BioMed Research International: Infectious Diseases The latest articles from Hindawi © 2017 , Hindawi Limited . All rights reserved. Analysis of a New Delayed HBV Model with Exposed State and Immune Response to Infected Cells and Viruses Thu, 16 Nov 2017 09:27:53 +0000 We propose a comprehensive delayed HBV model, which not only considers the immune response to both infected cells and viruses and a time delay for the immune system to clear viruses but also incorporates an exposed state and the proliferation of hepatocytes. We prove the positivity and boundedness of solutions and analyze the global stability of two boundary equilibria and then study the local asymptotic stability and Hopf bifurcation of the positive (infection) equilibrium and also the stability of the bifurcating periodic solutions. Moreover, we illustrate how the factors such as the time delay, the immune response to infected cells and viruses, and the proliferation of hepatocytes affect the dynamics of the model by numerical simulation. Deshun Sun and Fei Liu Copyright © 2017 Deshun Sun and Fei Liu. All rights reserved. Phylogenetic Diversity in Core Region of Hepatitis C Virus Genotype 1a as a Factor Associated with Fibrosis Severity in HIV-1-Coinfected Patients Sun, 12 Nov 2017 00:00:00 +0000 High hepatitis C virus (HCV) genetic diversity impacts infectivity/pathogenicity, influencing chronic liver disease progression associated with fibrosis degrees and hepatocellular carcinoma. HCV core protein is crucial in cell-growth regulation and host-gene expression. Liver fibrosis is accelerated by unknown mechanisms in human immunodeficiency virus-1- (HIV-1-) coinfected individuals. We aimed to study whether well-defined HCV-1a core polymorphisms and genetic heterogeneity are related to fibrosis in a highly homogeneous group of interferon-treated HIV-HCV-coinfected patients. Genetic heterogeneity was weighed by Faith’s phylogenetic diversity (PD), which has been little studied in HCV. Eighteen HCV/HIV-coinfected patients presenting different liver fibrosis stages before anti-HCV treatment-initiation were recruited. Sampling at baseline and during and after treatment was performed up to 72 weeks. At inter/intrahost level, HCV-1a populations were studied using molecular cloning and Sanger sequencing. Over 400 complete HCV-1a core sequences encompassing 573 positions of C were obtained. Amino acid substitutions found previously at positions 70 and 91 of HCV-1b core region were not observed. However, HCV genetic heterogeneity was higher in mild than in severe fibrosis cases. These results suggest a potential utility of PD as a virus-related factor associated with chronic hepatitis C progression. These observations should be reassessed in larger cohorts to corroborate our findings and assess other potential covariates. Micaela Parra, Natalia Laufer, Julieta M. Manrique, Leandro R. Jones, and Jorge Quarleri Copyright © 2017 Micaela Parra et al. All rights reserved. Transient Receptor Potential Melastatin 2 Negatively Regulates LPS-ATP-Induced Caspase-1-Dependent Pyroptosis of Bone Marrow-Derived Macrophage by Modulating ROS Production Wed, 08 Nov 2017 06:54:16 +0000 Background. Pyroptosis, a new form of cell death, which has special morphological characteristics, depends on caspase-1 activation and occupies an important role in inflammatory immune diseases and ischemia-reperfusion injury. ROS is a common activator of NLR/caspase-1. Transient receptor potential melastatin 2 (TRPM2), a selective cation channel, is involved in inflammatory regulation. This study was designed to explore the role of TRPM2 in activating caspase-1 and caspase-1-dependent pyroptosis of mouse BMDMs. Methods. BMDMs isolated from WT and TRPM2−/− mice were treated with LPS and ATP, along with ROS inhibitor (NAC and DPI), caspase-1 inhibitor (Z-YVAD), or not. The activation of caspase-1 was measured by western blot. EtBr and EthD-2 staining were used to assess the incidence of pyroptosis. Results. Compared with WT, the activated caspase-1-P10 was higher and the percentage of EtBr positive cells was also increased in TRPM2−/− group, which were both inhibited by Z-YVAD, NAC, or DPI. ASC oligomerization was increased in TRPM2−/− group. Conclusion. Deletion of TRPM2 can enhance the activation of caspase-1 and pyroptosis, which may be via modulating ROS production, suggesting that TRPM2 plays a critical role in immune adjustment. Haihong Wang, Xinyi Zhou, Hui Li, Xiaowei Qian, Yan Wang, and Liang Ma Copyright © 2017 Haihong Wang et al. All rights reserved. Relationship between Malaria and ABO Blood Types in East China Tue, 07 Nov 2017 07:19:50 +0000 Objectives. This study aims at investigating the relationship between malaria and blood group types in east China. Methods. Between 1 January 2011 and 31 March 2017, 99 malaria patients were enrolled for the study. Laboratory tests were conducted on their infection status and blood types. Clinical data of the participants were retrieved for analysis. Results. There was no mortality during the period of study. Overall, 90 (90.91%) of the patients were positive for Plasmodium falciparum, 8 (8.08%) were infected with Plasmodium vivax, and only 1 (1.01%) was infected with Plasmodium malariae. The most common blood group among the participants was group O (38.38%) followed by blood groups A, B, and AB, with 32.32%, 22.22%, and 7.07% cases, respectively. There was no significant relationship between the prevalence of malaria and ABO blood types (). In the blood group O, the prevalence of haemolytic-uremic syndrome and cerebral malaria was 13.16% and 5.25%, respectively, which was lower than that of the other three blood types (). Conclusion. There was no mortality among the malaria patients in this study. The blood group O was the most common blood type. Due to small sample size of data, there was no significant association between ABO blood types and malaria infection. Xuan Zhang, Meifang Yang, Hong Zhao, Jianhua Hu, and Lanjuan Li Copyright © 2017 Xuan Zhang et al. All rights reserved. Sodium Mercaptoethane Sulfonate Reduces Collagenolytic Degradation and Synergistically Enhances Antimicrobial Durability in an Antibiotic-Loaded Biopolymer Film for Prevention of Surgical-Site Infections Tue, 07 Nov 2017 00:00:00 +0000 Implant-associated surgical-site infections can have significant clinical consequences. Previously we reported a method for prophylactically disinfecting implant surfaces in surgical pockets, where an antibiotic solution containing minocycline (M) and rifampin (R) was applied as a solid film in a crosslinked biopolymer matrix that partially liquefied in situ to provide extended prophylaxis. Here we studied the effect of adding sodium 2-mercaptoethane sulfonate (MeSNA) on durability of prophylaxis in an in vitro model of implant-associated surgical-site infection. Adding MeSNA to the M/R biopolymer, antimicrobial film extended the duration for which biofilm formation by multidrug-resistant Pseudomonas aeruginosa (MDR-PA) was prevented on silicone surfaces in the model. M/R films with and without MeSNA were effective in preventing colonization by methicillin-resistant Staphylococcus aureus. Independent experiments revealed that MeSNA directly inhibited proteolytic digestion of the biopolymer film and synergistically enhanced antimicrobial potency of M/R against MDR-PA. Incubation of the MeSNA containing films with L929 fibroblasts revealed no impairment of cellular metabolic activity or viability. Joel Rosenblatt, Ruth A. Reitzel, George M. Viola, Nylev Vargas-Cruz, Jesse Selber, and Issam Raad Copyright © 2017 Joel Rosenblatt et al. All rights reserved. Sera from Visceral Leishmaniasis Patients Display Oxidative Activity and Affect the TNF-α Production by Macrophages In Vitro Sun, 05 Nov 2017 00:00:00 +0000 Mammalian protection against leishmanial infection depends on the development of an effective immune response. Zoonotic visceral leishmaniasis (ZVL) patients are usually unable to mount an effective immune response against the parasite and indeed appear to be severely immunosuppressed. This suppression has strong nonspecific and specific components mediated by serum factors and leishmanicidal activity of infected macrophages, respectively. The lipid profile has been shown to be altered in ZVL patients’ sera. This work aimed at (i) determining the HDL, Apo A1, LDL, and VLDL concentrations in ZVL patients’ sera; (ii) investigating the oxidative effect of ZVL patients’ sera on the β-carotene matrix; (iii) measuring IL-10, IL-6, IL-12p40, and tumour necrosis factor-α (TNF-α) concentrations in the macrophage cultures, to which 10% of ZVL patients’ serum had been added. Levels of HDL, LDL fraction, and apolipoprotein A1 in ZVL patients’ sera were lower than those of healthy individuals’ sera, except for the mean level of VLDL. The matrix of β-carotene and linoleic acid system was oxidized in the presence of ZLV patients’ sera. The presence of ZVL patients’ sera did not modify the cytokine production of IL-6, IL-12p40, and IL-10 by human macrophages in vitro but TNF-α production was altered, probably due to lack of macrophage stimulation by lipoprotein. Neci M. Soares, Joelma N. de Souza, Tatiana F. Leal, Eliana A. G. Reis, Maria S. Miranda, Washington L. C. dos Santos, and Márcia C. A. Teixeira Copyright © 2017 Neci M. Soares et al. All rights reserved. Prevalence of Helicobacter pylori vacA Genotypes and cagA Gene in Dental Plaque of Asymptomatic Mexican Children Wed, 01 Nov 2017 10:42:35 +0000 The variability in Helicobacter pylori vacA and cagA genes has been related to the progression of the gastrointestinal disease; also the presence of H. pylori in the oral cavity has been associated with periodontal disease in adults, but, in children without dyspeptic symptoms, little is known about this. We evaluated the prevalence of H. pylori and the presence of vacA/cagA genotypes in the oral cavity of Mexican children without dyspeptic symptoms. The gingival status was measured, and dental plaque samples () were taken. 38% of children were positive for H. pylori 16S rRNA gene by qPCR. A significant association between H. pylori oral infection and gingival status was observed (). In 34.6% (9/26) of mild gingivitis cases, s1m2 genotype was found, while s1m1 was typed in 50% (3/6) of moderate gingivitis. The cagA prevalence among H. pylori-positive children was 80.8% (21/26), 83.3% (5/6), and 16.7% (1/6) of cases of mild gingivitis, moderate gingivitis, and nongingivitis, respectively (). The s1m1/cagA+ combinational genotype was the most detected in children with gingivitis. Our results suggest that the prevalence of H. pylori and detection of vacA/cagA genotypes-associated gastrointestinal disease in the oral cavity could be related to the progression of gingivitis in asymptomatic children. Alejandra Mendoza-Cantú, Víctor Hugo Urrutia-Baca, Cynthia Sofía Urbina-Ríos, Myriam Angélica De la Garza-Ramos, Martha Elena García-Martínez, and Hilda H. H. Torre-Martínez Copyright © 2017 Alejandra Mendoza-Cantú et al. All rights reserved. Identification of Clostridium difficile Asymptomatic Carriers in a Tertiary Care Hospital Mon, 02 Oct 2017 08:58:40 +0000 Background. The diagnosis of Clostridium difficile infection (CDI) increases concern that asymptomatic carriers of toxigenic C. difficile may be diagnosed with CDI. Methods. A matched case control study was conducted in inpatients in a tertiary care center. The first 50 patients with diarrhea and a positive polymerase chain reaction (PCR) test beginning February 1, 2015, were identified as cases. Control patients were hospitalized patients receiving antibiotics, but with no diarrhea, housed in a room as close as possible to each case during the same admission time. A convenience sample of healthcare workers who cared for C. difficile infected patients was also tested. Results. We found two positive PCR results for C. difficile in controls (4.1%). None of these healthcare workers were positive for C. difficile by PCR. There was no difference between groups with respect to overall antibiotic use before the requested PCR for Clostridium difficile (). The majority of cases had a high proportion of gastrointestinal disorders (71.4%) compared with control (8.2%), . Patients with neoplasia had a higher chance of being identified as cases (). Conclusions. PCR should not be the only diagnostic tool but should be complementary to other methods and to the medical history. André Luiz de Oliveira Silva, Alexandre R. Marra, Marinês Dalla Valle Martino, Ana Carolina Cintra Nunes Mafra, Michael B. Edmond, and Oscar Fernando Pavao dos Santos Copyright © 2017 André Luiz de Oliveira Silva et al. All rights reserved. IFN-α Boosting of Mycobacterium bovis Bacillus Calmette Güerin-Vaccine Promoted Th1 Type Cellular Response and Protection against M. tuberculosis Infection Wed, 27 Sep 2017 00:00:00 +0000 The role of type I IFNs in the pathogenesis and control of mycobacterial infection is still controversial. It has been reported that type I IFNs exacerbated M. tuberculosis infection through hampering Th1 type cellular immune response. However, under certain conditions they can act as natural immune adjuvants for commercial vaccines. At this point, we have reported recently that successive IFN-alpha boosting of Mycobacterium bovis Bacillus Calmette Güerin (BCG) vaccinated mice protected adult mice from intradermal M. lepraemurium infection and a difference in iNOS was observed. In the present work, we have found that intramuscular IFN-α boosting of Mycobacterium bovis Bacillus Calmette Güerin (BCG) vaccine, either in vitro (human cell line or macrophages derived from PBMC) or in vivo (aerosol mouse model of MTb infection), promoted mostly the development of specific anti-antimycobacterial Th1 type cytokines (IFN-γ; IL-12, TNF-alpha, and IL-17; IL1β) while bacterial load reduction (0.9 logs versus PBS or BCG vaccine) was observed. These findings indicate that, under the experimental settings reported here, interferon alpha can drive or affect the TH cellular immune response in favour of BCG-inducing immunity against M. tuberculosis infection. C. E. Rivas-Santiago and G. G. Guerrero Copyright © 2017 C. E. Rivas-Santiago and G. G. Guerrero. All rights reserved. Comment on “Ebola Virus Infection among Western Healthcare Workers Unable to Recall the Transmission Route” Wed, 20 Sep 2017 00:00:00 +0000 Mark G. Kortepeter, Theodore J. Cieslak, Elena H. Kwon, Philip W. Smith, Christopher J. Kratochvil, and Angela L. Hewlett Copyright © 2017 Mark G. Kortepeter et al. All rights reserved. Improving Prediction Accuracy of “Central Line-Associated Blood Stream Infections” Using Data Mining Models Wed, 20 Sep 2017 00:00:00 +0000 Prediction of nosocomial infections among patients is an important part of clinical surveillance programs to enable the related personnel to take preventive actions in advance. Designing a clinical surveillance program with capability of predicting nosocomial infections is a challenging task due to several reasons, including high dimensionality of medical data, heterogenous data representation, and special knowledge required to extract patterns for prediction. In this paper, we present details of six data mining methods implemented using cross industry standard process for data mining to predict central line-associated blood stream infections. For our study, we selected datasets of healthcare-associated infections from US National Healthcare Safety Network and consumer survey data from Hospital Consumer Assessment of Healthcare Providers and Systems. Our experiments show that central line-associated blood stream infections (CLABSIs) can be successfully predicted using AdaBoost method with an accuracy up to 89.7%. This will help in implementing effective clinical surveillance programs for infection control, as well as improving the accuracy detection of CLABSIs. Also, this reduces patients’ hospital stay cost and maintains patients’ safety. Amin Y. Noaman, Farrukh Nadeem, Abdul Hamid M. Ragab, Arwa Jamjoom, Nabeela Al-Abdullah, Mahreen Nasir, and Anser G. Ali Copyright © 2017 Amin Y. Noaman et al. All rights reserved. Sterilization of Biofilm on a Titanium Surface Using a Combination of Nonthermal Plasma and Chlorhexidine Digluconate Tue, 19 Sep 2017 09:31:11 +0000 Nosocomial infections caused by opportunistic bacteria pose major healthcare problem worldwide. Out of the many microorganisms responsible for such infections, Pseudomonas aeruginosa is a ubiquitous bacterium that accounts for 10–20% of hospital-acquired infections. These infections have mortality rates ranging from 18 to 60% and the cost of treatment ranges from $20,000 to $80,000 per infection. The formation of biofilms on medical devices and implants is responsible for the majority of those infections. Only limited progress has been made to prevent this issue in a safe and cost-effective manner. To address this, we propose employing jet plasma to break down and inactivate biofilms in vitro. Moreover, to improve the antimicrobial effect on the biofilm, a treatment method using a combination of jet plasma and a biocide known as chlorhexidine (CHX) digluconate was investigated. We found that complete sterilization of P. aeruginosa biofilms can be achieved after combinatorial treatment using plasma and CHX. A decrease in biofilm viability was also observed using confocal laser scanning electron microscopy (CLSM). This treatment method sterilized biofilm-contaminated surfaces in a short treatment time, indicating it to be a potential tool for the removal of biofilms present on medical devices and implants. Tripti Thapa Gupta, Surya B. Karki, Jyl S. Matson, Daniel J. Gehling, and Halim Ayan Copyright © 2017 Tripti Thapa Gupta et al. All rights reserved. The Clinical Significance of FilmArray Respiratory Panel in Diagnosing Community-Acquired Pneumonia Wed, 06 Sep 2017 07:22:23 +0000 Aim. FilmArray Respiratory Panel (FilmArray RP) test is an emerging diagnostic method in fast detecting multiple respiratory pathogens; the methodology and clinical significance of FilmArray RP in community-acquired pneumonia (CAP) diagnosis were evaluated in this study. Methods. Specimens from 74 patients with CAP were analyzed and compared using FilmArray RP, traditional multiple PCR assay, bacterial (or fungal) culture, and serological detection. Results. FilmArray RP and multiplex PCR showed 100% coincidence rate in detecting coronaviruses 229E, OC43, HKU1, and NL63, human metapneumovirus, influenza A and B, and parainfluenza viruses (PIV1, PIV2, and PIV4). There were 15 viral specimens tested as disagreement positive results. FilmArray RP had higher detection rate in detecting dual viral and Mycoplasma pneumoniae infection. The positive bacteria (or fungi) were found in 25 specimens. Conclusions. This study demonstrated the capability of FilmArray RP for simultaneous detection of broad-spectrum respiratory pathogens and potential use in facilitating better patient care. Huanzhu Chen, Huilan Weng, Meirui Lin, Ping He, Yazhen Li, Qingdong Xie, Changwen Ke, and Xiaoyang Jiao Copyright © 2017 Huanzhu Chen et al. All rights reserved. CT Scanning in Identification of Sheep Cystic Echinococcosis Thu, 31 Aug 2017 00:00:00 +0000 Objective. We aim to determine the efficiency of CT in identification of cystic echinococcosis in sheep. Methods. Fifty-three sheep with liver cysts confirmed by ultrasonography were subject to CT scan to evaluate the number, size, and type of the cysts in liver and lung, confirmed using necropsy. The correlation of numbers between liver cysts and lung cysts was calculated using Pearson analysis. Results. Necropsy indicated a 98% consensus on size, location, number, and activity compared with CT scan. The viable cysts were 53.1% and 50.6% in the liver and lung, respectively. Among the cysts in liver, 35.5%, 9.5%, 5.7%, 10.2%, and 39.1% were Types CE1, CE2, CE3, CE4, and CE5, respectively. The cysts in the lungs, 17.4%, 26.9%, 12.1%, 11.6%, and 32.1%, were Types CE1, CE2, CE3, CE4, and CE5, respectively. A significant correlation was noticed between the number of cysts in liver and those in lung (, ). Conclusions. CT scan is a suitable tool in determining the size and type of cystic hydatid cysts in both liver and lung of sheep. A significant correlation was noticed between the numbers in liver and lung, indicating that lung infection was likely due to the expansion of liver cyst burden pressure. Rui Mao, Hongzhi Qi, Lei Pei, Jie Hao, Jian Dong, Tao Jiang, Abudula Ainiwaer, Ge Shang, Lin Xu, Xi Shou, Songan Zhang, Ge Wu, Pengfei Lu, Yongxing Bao, and Haitao Li Copyright © 2017 Rui Mao et al. All rights reserved. Isolation, Identification, and Antimicrobial Susceptibility of Bacteria Associated with Waterpipe Contaminants in Selected Area of Saudi Arabia Mon, 28 Aug 2017 09:56:09 +0000 This study highlights the level of microbial contamination of waterpipe components in selected area of Saudi Arabia and the resistance of selected bacteria to different antibiotics was determined. A series of biochemical tests, microscopic examination, and screening on Vitek 2 compact (bioMérieux Inc., USA) system were done to characterize the bacterial isolates. Out of 132 samples investigated, 7 mouthpiece samples and 48 water bowl samples showed positivity on culture. The percentage of contamination rate was higher in water bowl (69.69%) than in mouthpieces (10.6%) for all selected areas. A total of 55 bacterial isolates were identified which included Gram-negative (28) and Gram-positive (27) bacteria. Antimicrobial susceptibility data showed more resistance to bacteria isolated from water bowl than bacteria isolated from mouthpiece. In addition, one isolate which was confirmed as methicillin-resistant Staphylococcus aureus and Klebsiella pneumoniae was resistant to antibiotics which are commonly used to treat pneumonia. Water bowl of waterpipe instrument is significantly contaminated with different bacterial pathogens including multidrug-resistant and pneumonia causing bacteria, which are a real health concern among waterpipe smokers. The presented data could assist public health professionals to raise the concerns regarding cleaning practices of waterpipe components and highlights the risk posed among the waterpipe smokers. Mohammed Alaidarous, Meshal Alanazi, and Ahmed Abdel-Hadi Copyright © 2017 Mohammed Alaidarous et al. All rights reserved. Acute Bacterial Meningitis in Qatar: A Hospital-Based Study from 2009 to 2013 Thu, 13 Jul 2017 00:00:00 +0000 Background and Objectives. Bacterial meningitis is a common medical condition in Qatar. The aim of this study was to describe the clinical characteristics of bacterial meningitis, the frequency of each pathogen, and its sensitivity to antibiotics and risk factors for death. Patients and Methods. This retrospective study was conducted at Hamad General Hospital between January 1, 2009, and December 31, 2013. Results. We identified 117 episodes of acute bacterial meningitis in 110 patients. Their mean age was years (range: 2–74) and 81 (69.2%) of them were male patients. Fifty-nine episodes (50.4%) were community-acquired infection and fever was the most frequent symptom (94%), whereas neurosurgery is the most common underlying condition. Coagulase-negative staphylococci were the most common causative agent, of which 95% were oxacillin-resistant, while 63.3% of Acinetobacter spp. showed resistance to meropenem. The in-hospital mortality was 14 (12%). Only the presence of underlying diseases, hypotension, and inappropriate treatment were found to be independent predictors of mortality. Conclusion. Acute bacterial meningitis predominantly affected adults and coagulase-negative staphylococci species were the common causative agent in Qatar with majority of infections occurring nosocomially. More than 90% of all implicated coagulase-negative staphylococci strains were oxacillin-resistant. Fahmi Yousef Khan, Mohammed Abu-Khattab, Eman Abdulrahman Almaslamani, Abubaker Ahmed Hassan, Shehab Fareed Mohamed, Abdurrahman Ali Elbuzdi, Nada Yagoub Elmaki, Deshmukh Anand, and Doiphode Sanjay Copyright © 2017 Fahmi Yousef Khan et al. All rights reserved. The Effects of Time Lag and Cure Rate on the Global Dynamics of HIV-1 Model Tue, 13 Jun 2017 10:05:15 +0000 In this research article, a new mathematical model of delayed differential equations is developed which discusses the interaction among CD T cells, human immunodeficiency virus (HIV), and recombinant virus with cure rate. The model has two distributed intracellular delays. These delays denote the time needed for the infection of a cell. The dynamics of the model are completely described by the basic reproduction numbers represented by , , and . It is shown that if , then the infection-free equilibrium is locally as well as globally stable. Similarly, it is proved that the recombinant absent equilibrium is locally as well as globally asymptotically stable if . Finally, numerical simulations are presented to illustrate our theoretical results. Our obtained results show that intracellular delay and cure rate have a positive role in the reduction of infected cells and the increasing of uninfected cells due to which the infection is reduced. Nigar Ali, Gul Zaman, Abdullah, Aisha M. Alqahtani, and Ali Saleh Alshomrani Copyright © 2017 Nigar Ali et al. All rights reserved. Chlamydia muridarum Infection of Macrophages Stimulates IL-1β Secretion and Cell Death via Activation of Caspase-1 in an RIP3-Independent Manner Sun, 04 Jun 2017 00:00:00 +0000 Chlamydiae are Gram-negative bacteria, which replicate exclusively in the infected host cells. Infection of the host cells by Chlamydiae stimulates the innate immune system leading to an inflammatory response, which is manifested not only by secretion of proinflammatory cytokines such as IL-1 from monocytes, macrophages, and dendritic cells, but also possibly by cell death mediated by Caspase-1 pyroptosis. RIP3 is a molecular switch that determines the development of necrosis or inflammation. However, the involvement of RIP3 in inflammasome activation by Chlamydia muridarum infection has not been clarified. Here, we assessed the role of RIP3 in synergy with Caspase-1 in the induction of IL-1β production in BMDM after either LPS/ATP or Chlamydia muridarum stimulation. The possibility of pyroptosis and necroptosis interplays and the role of RIP3 in IL-1 production during Chlamydia muridarum infection in BMDM was investigated as well. The data indicated that RIP3 is involved in NLRP3 inflammasome activation in LPS/ATP-stimulated BMDMs but not in Chlamydia muridarum infection. Pyroptosis occurred in BMDM after LPS/ATP stimulation or Chlamydia muridarum infection. Moreover, the results also illuminated the important role of the Caspase-1-mediated pyroptosis process which does not involve RIP3. Taken together, these observations may help shed new light on details in inflammatory signaling pathways activated by Chlamydia muridarum infection. Lixiang Chen, Xue Liu, Xin Yu, Rongrong Ren, Chao Wang, Rui Zhao, Guangxun Meng, Shun Li, and Xiaohui Zhou Copyright © 2017 Lixiang Chen et al. All rights reserved. Involvement of the Toll-Like Receptor/Nitric Oxide Signaling Pathway in the Pathogenesis of Cervical Cancer Caused by High-Risk Human Papillomavirus Infection Wed, 24 May 2017 00:00:00 +0000 Human papillomavirus (HPV) can activate Toll-like receptor (TLR)/nitric oxide (NO) signaling pathways; however, whether the TLR/NO pathway is involved in cervical cancer caused by high-risk HPV (HR-HPV) remains unclear. In this study, 43 HR-HPV-positive patients with cervical cancer (CC group), 39 HR-HPV-positive patients with a healthy cervix (HR-HPV group), and 33 HR-HPV-negative controls were recruited. NO concentration in cervical canal and expression of inducible NO synthase (iNOS) in cervical tissues were detected. Expressions of key TLR/NO pathway genes (TLR3/4/7/8, NF-κB p65, and iNOS) in cervical epithelial cells were detected by quantitative reverse transcription PCR. Expressions of TLR4, NF-κB p65, and iNOS in CaSki, HeLa, and C33a cells were determined by Western blot. NO concentration in cervical canal of CC group was significantly higher than in other groups (). Positive rates of iNOS in cervical tissues were 72.1%, 28.2%, and 3.1% in the CC group, HR-HPV group, and controls, respectively (). Levels of TLR3, TLR4, TLR7, TLR8, NF-κB p65, and iNOS in cervical epithelial cells were higher in CC group than in other groups (). Both mRNA and protein levels of TLR4, NF-κB p65, and iNOS were higher in HPV-positive HeLa and CaSki cells than in HPV-negative C33a cells (). Together, these results suggest that TLR/NO signaling pathway may be involved in pathogenesis of cervical cancer caused by HR-HPV. Jie Li, Heping Rao, Chang’e Jin, and Jinrong Liu Copyright © 2017 Jie Li et al. All rights reserved. First Insight into the Molecular Epidemiology of Mycobacterium tuberculosis Isolates from the Minority Enclaves of Southwestern China Wed, 17 May 2017 09:29:31 +0000 Yunnan is a province located in southwestern China. The aim of this study was to investigate the genetic diversity of Mycobacterium tuberculosis (MTB) strains circulating in Yunnan Province. We used spoligotyping and a 12-locus high-resolution VNTR set to identify a total of 271 MTB isolates collected from six prefectures’ Yunnan. All the 271 patients were classified as 11 different ethnic groups, including 133 Han patients (49.1%) and 138 minority patients (50.9%). Spoligotyping analyses revealed that the largest two spoligotypes were SIT1 (Beijing family, ) and SIT53 (T family, ). Statistical analysis indicated that the proportion of Beijing genotype in Qujing was significantly higher than that in the ethnic enclaves (). Compared with the percentage of Beijing-family isolates from patients under 25 years of age (72.7%), those from patients aged 45~64 years (44.6%, ) and older than 64 years (48.1%) () were significantly lower. Beijing genotype strains (23.8%, 36/151) showed higher clustering rate than non-Beijing genotype strains (16/120, 13.3%, ). In conclusion, our data demonstrated that the Beijing genotype was the predominant genotype in Yunnan Province. The distribution of Beijing genotype strains showed geographic diversity. In addition, Beijing genotype was associated with recent transmission rather than drug resistance. Lianyong Chen, Yu Pang, Li Ma, Huijuan Yang, Haohao Ru, Xing Yang, Shuangqun Yan, Mao Jia, and Lin Xu Copyright © 2017 Lianyong Chen et al. All rights reserved. Analysis of Differentially Expressed Proteins in Mycobacterium avium-Infected Macrophages Comparing with Mycobacterium tuberculosis-Infected Macrophages Wed, 10 May 2017 00:00:00 +0000 Mycobacterium avium (MA) belongs to the intracellular parasitic bacteria. To better understand how MA survives within macrophages and the different pathogenic mechanisms of MA and Mycobacterium tuberculosis (MTB), tandem mass tag (TMT) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis have been used to determine the proteins which are differentially expressed in MA-infected and MTB-infected macrophages. 369 proteins were found to be differentially expressed in MA-infected cells but not in MTB-infected cells. By using certain bioinformatics methods, we found the 369 proteins were involved in molecular function, biological process, and cellular component including binding, catalytic activity, metabolic process, cellular process, and cell part. In addition, some identified proteins were involved in multiple signaling pathways. These results suggest that MA probably survive within macrophages by affecting the expression of some crucial proteins. Dongjun Yang, Xin Fu, Shiyi He, Xueping Ning, and Min Ling Copyright © 2017 Dongjun Yang et al. All rights reserved. Positive Correlation between IP-10 and IFN-γ Levels in Rhesus Monkeys (Macaca mulatta) with Either Naturally Acquired or Experimental Infection of Mycobacterium tuberculosis Thu, 20 Apr 2017 07:47:36 +0000 Numerous studies identify that IP-10 and IFN-γ are involved in leucocyte migration and activation and regarded as promising surrogate biomarkers in human and bovine tuberculosis infection, but there is lack of evidence for IP-10 in nonhuman primates. In this study, we directly determined IP-10 and IFN-γ levels in plasma from 30 healthy monkeys, 30 monkeys with naturally acquired tuberculosis, 4 monkeys experimentally infected with tuberculosis, and PPD stimulated whole blood of 14 monkeys with naturally acquired tuberculosis by ELISA. Higher plasma levels of IP-10 and IFN-γ were observed in natural tuberculosis monkeys than in healthy controls. The dynamic changes of plasma IP-10 and IFN-γ in experimental infections showed consistent representation of a transient increase during the infection period. After PPD stimulation, release of IP-10 and IFN-γ is significantly induced in natural tuberculosis monkeys, but the stimulation index of IP-10 was significantly lower than IFN-γ. Further analysis showed that positive correlation between IP-10 and IFN-γ existed in healthy and tuberculosis monkeys. Our findings support plasma IP-10 and IFN-γ as biomarkers for monitoring ongoing inflammation of nonhuman primate tuberculosis, and IFN-γ is a more valuable diagnostic biomarker. Fangui Min, Ruike Wu, Jinchun Pan, Shuwu Huang, Yinzhu Luo, and Yu Zhang Copyright © 2017 Fangui Min et al. All rights reserved. Drug Susceptibility of 33 Reference Strains of Slowly Growing Mycobacteria to 19 Antimicrobial Agents Thu, 20 Apr 2017 07:27:47 +0000 Objectives. Slowly growing mycobacteria (SGM) are prevalent worldwide and cause an extensive spectrum of diseases. Methods. In this study, the antimicrobial susceptibility of 33 reference strains of SGM to 19 antimicrobial agents was tested using a modified microdilution method. Results. Cefmetazole (32/33) and azithromycin (32/33) exhibited the highest antimicrobial activity, and dapsone (9/33) exhibited the lowest activity against the tested strains. Cefoxitin (30/33), cefoperazone (28/33), and cefepime (28/33) were effective against a high proportion of strains, and macrolides were also highly effective as well as offering the benefit of convenient oral administration to patients. Linezolid (27/33), meropenem (26/33), sulfamethoxazole (26/33), and tigecycline (25/33) showed the highest activity; clofazimine (20/33) and doxycycline (18/33) showed intermediate activity; and rifapentine (13/33), rifabutin (13/33), and minocycline (11/33) showed low antimicrobial activity, closely followed by thioacetazone (10/33) and pasiniazid (10/33), against the tested organisms. According to their susceptibility profiles, the slowly growing species Mycobacterium avium and Mycobacterium simiae were the least susceptible to the tested drugs, whereas Mycobacterium intracellulare, Mycobacterium asiaticum, Mycobacterium scrofulaceum, Mycobacterium szulgai, Mycobacterium branderi, and Mycobacterium holsaticum were the most susceptible. Conclusions. In summary, cephalosporins and macrolides, particularly cefmetazole, azithromycin, clarithromycin, and roxithromycin, showed good antimicrobial activity against the reference strains of SGM. Hui Pang, Yi Jiang, and Kanglin Wan Copyright © 2017 Hui Pang et al. All rights reserved. Etiological and Resistance Profile of Bacteria Involved in Urinary Tract Infections in Young Children Tue, 11 Apr 2017 07:04:16 +0000 Background. The objective of this study was to identify the bacteria most frequently responsible for urinary tract infection (UTI) in the population of under-2-year-olds in our geographic area and to evaluate the activity of antibiotics widely used for UTI treatment during a 4-year study period. Materials and Methods. A retrospective analysis was conducted of data on the identification and susceptibility of microorganisms isolated in urine samples from children under 2 years of age. Results. A total of 1,045 uropathogens were isolated. Escherichia coli accounted for the majority (60.3%) of these, followed by Enterococcus faecalis (22.4%) and Klebsiella spp. (6.5%). The highest E. coli susceptibility rates (>90%) were to piperacillin-tazobactam, cefuroxime, cefotaxime, ceftazidime, imipenem, gentamicin, nitrofurantoin, and fosfomycin, and the lowest were to amoxicillin-clavulanic acid and cotrimoxazole. Among all bacteria isolated, we highlight the overall high activity of piperacillin-tazobactam, imipenem, nitrofurantoin, and fosfomycin against both community and hospital isolates and the reduced activity of amoxicillin-clavulanic acid, cephalosporins, gentamicin, and cotrimoxazole. There was no significant change in the total activity of any of the studied antibiotics over the 4-year study period. Conclusion. Empiric treatment with amoxicillin-clavulanic acid, cotrimoxazole, cephalosporins, and gentamicin may be inadequate due to their limited activity against uropathogens in our setting. Antonio Sorlózano-Puerto, José María Gómez-Luque, Juan de Dios Luna-del-Castillo, José María Navarro-Marí, and José Gutiérrez-Fernández Copyright © 2017 Antonio Sorlózano-Puerto et al. All rights reserved. The Interaction between HIV and Intestinal Helminth Parasites Coinfection with Nutrition among Adults in KwaZulu-Natal, South Africa Wed, 22 Mar 2017 10:04:10 +0000 In South Africa few studies have examined the effects of the overlap of HIV and helminth infections on nutritional status. This cross-sectional study investigated the interaction between HIV and intestinal helminths coinfection with nutritional status among KwaZulu-Natal adults. Participants were recruited from a comprehensive primary health care clinic and stratified based on their HIV, stool parasitology, IgE, and IgG4 results into four groups: the uninfected, HIV infected, helminth infected, and HIV-helminth coinfected groups. The nutritional status was assessed using body mass index, 24-hour food recall, micro-, and macronutrient biochemical markers. Univariate and multivariate multinomial probit regression models were used to assess nutritional factors associated with singly and dually infected groups using the uninfected group as a reference category. Biochemically, the HIV-helminth coinfected group was associated with a significantly higher total protein, higher percentage of transferrin saturation, and significantly lower ferritin. There was no significant association between single or dual infections with HIV and helminths with micro- and macronutrient deficiency; however general obesity and low micronutrient intake patterns, which may indicate a general predisposition to micronutrient and protein-energy deficiency, were observed and may need further investigations. B. T. Mkhize, M. Mabaso, T. Mamba, C. E. Napier, and Z. L. Mkhize-Kwitshana Copyright © 2017 B. T. Mkhize et al. All rights reserved. Clinical Features and Outcome in Adult Cases of Tuberculous Meningitis in Tertiary Care Hospital in Antananarivo, Madagascar Wed, 15 Mar 2017 00:00:00 +0000 Purpose. We aimed to describe and to assess prognosis factors in tuberculous meningitis in adult patients. Methods. We performed a retrospective study of case records of adult patients. Patients classified as definite, probable, or possible tuberculous meningitis according to standardized definition criteria were included and assessed in the study. Results. Seventy-five patients were included in the study. Tuberculous meningitis was classified as definite in 8 (10.7%), probable in 44 (58.7%), and possible in 23 patients (30.6%). HIV was found in 3% of patients. Patients were in advanced stages at admission in 82.7%. Median duration of symptoms prior to admission was 3 weeks (IQR: 2–5). Median time to diagnosis following admission was 5 days (IQR: 3–8). Median CSF WCC was 75 per mm3 with lymphocytic predominance in 38 cases (52.8%). Median CSF glucose level was 1.48 mmol/L and median CSF protein level was 1 g/L. Mortality rate was 28%. Age ≥ 35 years (aOR: 4.06; 95% CI: 1.16–14.26) and coma (aOR: 12.98; 95% CI: 1.13–149.16) predicted inpatient mortality. Conclusion. Most of the patients experienced more than 3 weeks of diagnostic delay prior to admission. Mortality was high and occurred early after admission. Age and coma were identified as independent prognosis factors. Mihaja Raberahona, Rivonirina Andry Rakotoarivelo, Tiana Razafinambinintsoa, Radonirina Lazasoa Andrianasolo, and Mamy Jean de Dieu Randria Copyright © 2017 Mihaja Raberahona et al. All rights reserved. Potential Role of Carvedilol in the Cardiac Immune Response Induced by Experimental Infection with Trypanosoma cruzi Thu, 09 Mar 2017 00:00:00 +0000 Trypanosoma cruzi causes a cardiac infection characterized by an inflammatory imbalance that could become the inciting factor of the illness. To this end, we evaluated the role of carvedilol, a beta-blocker with potential immunomodulatory properties, on the immune response in C57BL/6 mice infected with VL-10 strain of T. cruzi in the acute phase. Animals () were grouped: (i) not infected, (ii) infected, (iii) infected + carvedilol, and (iv) not infected + carvedilol. We analyzed parameters related to parasitemia, plasma levels of TNF, IL-10, and CCL2, and cardiac histopathology after the administration of carvedilol for 30 days. We did not observe differences in the maximum peaks of parasitemia in the day of their detection among the groups. The plasma TNF was elevated at 60 days of infection in mice treated or not with carvedilol. However, we observed a decreased CCL2 level and increased IL-10 levels in those infected animals treated with carvedilol, which impacted the reduction of the inflammatory infiltration in cardiac tissue. For this experimental model, carvedilol therapy was not able to alter the levels of circulating parasites but modulates the pattern of CCL2 and IL-10 mediators when the VL10 strain of T. cruzi was used in C57BL6 mice. Aline Luciano Horta, Ana Luisa Junqueira Leite, G. Paula Costa, Vivian Paulino Figueiredo, and André Talvani Copyright © 2017 Aline Luciano Horta et al. All rights reserved. Effect of Prior Atorvastatin Treatment on the Frequency of Hospital Acquired Pneumonia and Evolution of Biomarkers in Patients with Acute Ischemic Stroke: A Multicenter Prospective Study Sun, 05 Mar 2017 00:00:00 +0000 Objective. To investigate whether prior treatment of atorvastatin reduces the frequency of hospital acquired pneumonia (HAP). Methods. Totally, 492 patients with acute ischemic stroke and Glasgow Coma Scale ≤ 8 were enrolled in this study. Subjects were assigned to prior atorvastatin treatment group (, PG) and no prior treatment group (, NG). All the patients were given 20 mg atorvastatin every night during their hospital stay. HAP frequency and 28-day mortality were measured. Levels of inflammatory biomarkers [white blood cell (WBC), procalcitonin (PCT), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6)] were tested. Results. There was no significant difference in the incidence of HAP between PG and NG (25.74% versus. 24.55%, ) and 28-day mortality (50.72% versus 58.18%, ). However, prior statin treatment did modify the mortality of ventilator associated pneumonia (VAP) (36.54% versus 58.14%, ) and proved to be a protective factor (HR, 0.564; 95% CI, 0.310~0.825, ). Concentrations of TNF-α and IL-6 in PG VAP cases were lower than those in NG VAP cases (). Conclusions. Prior atorvastatin treatment in patients with ischemic stroke was associated with a lower concentration of IL-6 and TNF-α and improved the outcome of VAP. This clinical study has been registered with ChiCTR-ROC-17010633 in Chinese Clinical Trial Registry. Yuetian Yu, Cheng Zhu, Chunyan Liu, and Yuan Gao Copyright © 2017 Yuetian Yu et al. All rights reserved. Screening for Tuberculosis in Health Care Workers: Experience in an Italian Teaching Hospital Mon, 27 Feb 2017 06:35:53 +0000 Health care workers (HCW) are particularly at risk of acquiring tuberculosis (TB), even in countries with low TB incidence. Therefore, TB screening in HCW is a useful prevention strategy in countries with both low and high TB incidence. Tuberculin skin test (TST) is widely used although it suffers of low specificity; on the contrary, the in vitro enzyme immunoassay tests (IGRA) show superior specificity and sensitivity but are more expensive. The present study reports the results of a three-year TB surveillance among HCW in a large teaching hospital in Rome, using TST (by standard Mantoux technique) and IGRA (by QuantiFERON-TB) as first- and second-level screening tests, respectively. Out of 2290 HCW enrolled, 141 (6.1%) had a positive TST; among them, 99 (70.2%) underwent the IGRA and 16 tested positive (16.1%). The frequency of HCW tested positive for TB seems not far from other experiences in low incidence countries. Our results confirm the higher specificity of IGRA, but, due to its higher cost, TST can be considered a good first level screening test, whose positive results should be further confirmed by IGRA before the patients undergo X-ray diagnosis and/or chemotherapy. Christian Napoli, Filippo Ferretti, Filippo Di Ninno, Riccardo Orioli, Alessandra Marani, Maria Giuditta Sarlo, Claudio Prestigiacomo, Assunta De Luca, and Giovanni Battista Orsi Copyright © 2017 Christian Napoli et al. All rights reserved. IFN-τ Displays Anti-Inflammatory Effects on Staphylococcus aureus Endometritis via Inhibiting the Activation of the NF-κB and MAPK Pathways in Mice Sun, 26 Feb 2017 00:00:00 +0000 The aim of the present study was to determine the anti-inflammatory effect of IFN-τ on endometritis using a mouse model of S. aureus-induced endometritis and to elucidate the mechanism of action underlying these effects. In the present study, the effect of IFN-τ on S. aureus growth was monitored by turbidimeter at 600 nm. IFN-τ did not affect S. aureus growth. The histopathological changes indicated that IFN-τ had a protective effect on uterus tissues with S. aureus infection. The ELISA and qPCR results showed the production of the proinflammatory cytokines TNF-α, IL-1β, and IL-6 was decreased with IFN-τ treatment. In contrast, the level of the anti-inflammatory cytokine IL-10 was increased. We further studied the signaling pathway associated with these observations, and the qPCR results showed that the expression of TLR2 was repressed by IFN-τ. Furthermore, the western blotting results showed the phosphorylation of IκB, NF-κB p65, and MAPKs (p38, JNK, and ERK) was inhibited by IFN-τ treatment. The results suggested that IFN-τ may be a potential drug for the treatment of uterine infection due to S. aureus or other infectious inflammatory diseases. Zhenbiao Zhang, Yingfang Guo, Yuzhu Liu, Chengye Li, Mengyao Guo, and Ganzhen Deng Copyright © 2017 Zhenbiao Zhang et al. All rights reserved. Interferon-Gamma Release Assay Performance of Cerebrospinal Fluid and Peripheral Blood in Tuberculous Meningitis in China Mon, 20 Feb 2017 00:00:00 +0000 The aim of this study was to examine the performance of T-SPOT.TB on cerebrospinal fluid (CSF) and peripheral blood (PB) in diagnosis of tuberculous meningitis (TBM) in China. Of 100 patients with presumed TBM prospectively enrolled from Sep 2012 to Oct 2014, 53 were TBM (21 definite and 32 probable TBM cases) and 37 were non-TBM cases; the other 10 patients were excluded from analysis due to inconclusive diagnosis, no sufficient CSF samples, or incomplete follow-up. T-SPOT.TB on CSF and PB and routine laboratory tests of CSF were performed simultaneously. The receiver operating characteristic (ROC) curve and cut-off value of CSF T-SPOT.TB and routine CSF parameters were established between TBM and non-TBM group. The area under ROC curve (AUC) of the T-SPOT.TB on CSF and PB was 0.81 and 0.89, which was higher than that of the routine CSF parameters (AUC 0.67–0.77). Although the sensitivity of CSF T-SPOT.TB was lower than that of PB T-SPOT.TB (60.8% versus 90.6%, ), the specificity of CSF T-SPOT.TB was significantly higher than that of PB T-SPOT.TB (97.2% versus 75.7%, ). These results indicated that the diagnostic accuracies of PB and CSF T-SPOT.TB are higher than routine laboratory tests. Furthermore, the higher specificity of CSF T-SPOT.TB makes it a useful rule-in test in rapid diagnosis of TBM. Liping Pan, Fei Liu, Jinli Zhang, Xinting Yang, Shiqi Zheng, Jing Li, Hongyan Jia, Xiaoyou Chen, Mengqiu Gao, and Zongde Zhang Copyright © 2017 Liping Pan et al. All rights reserved. Emerging and Neglected Infectious Diseases: Insights, Advances, and Challenges Mon, 13 Feb 2017 08:20:54 +0000 Infectious diseases are a significant burden on public health and economic stability of societies all over the world. They have for centuries been among the leading causes of death and disability and presented growing challenges to health security and human progress. The threat posed by infectious diseases is further deepened by the continued emergence of new, unrecognized, and old infectious disease epidemics of global impact. Over the past three and half decades at least 30 new infectious agents affecting humans have emerged, most of which are zoonotic and their origins have been shown to correlate significantly with socioeconomic, environmental, and ecological factors. As these factors continue to increase, putting people in increased contact with the disease causing pathogens, there is concern that infectious diseases may continue to present a formidable challenge. Constant awareness and pursuance of effective strategies for controlling infectious diseases and disease emergence thus remain crucial. This review presents current updates on emerging and neglected infectious diseases and highlights the scope, dynamics, and advances in infectious disease management with particular focus on WHO top priority emerging infectious diseases (EIDs) and neglected tropical infectious diseases. Nicholas Israel Nii-Trebi Copyright © 2017 Nicholas Israel Nii-Trebi. All rights reserved. Advances in Emerging and Neglected Infectious Diseases Mon, 30 Jan 2017 08:32:01 +0000 Charles T. Spencer and Jose Ronnie Vasconcelos Copyright © 2017 Charles T. Spencer and Jose Ronnie Vasconcelos. All rights reserved. Prevalence of Hepatitis C Virus Infection and Its Risk Factors among Patients Attending Rwanda Military Hospital, Rwanda Thu, 26 Jan 2017 00:00:00 +0000 In Rwanda, the prevalence of viral hepatitis (HCV) is poorly understood. The current study investigated the prevalence and risk factors of HCV infection in Rwanda. A total of 324 patients attending Rwanda Military Hospital were randomly selected and a questionnaire was administered to determine the risk factors. Blood was collected and screened for anti-HCV antibodies and seropositive samples were subjected to polymerase chain reaction method. Hematology abnormalities in the HCV infected patients were also investigated. Anti-HCV antibody and active HCV infection were found in 16.0% and 9.6% of total participants, respectively. Prevalence was highest (28.4%; 19/67) among participants above 55 years and least (2.4%; 3/123) among younger participants (18–35 years). There was a significant () relationship between place of residence and HCV infection with residents of Southern Province having significantly higher prevalence. The hematological abnormalities observed in the HCV infected patients included leukopenia (48.4%; 15/52), neutropenia (6.5%; 2/52), and thrombocytopenia (25.8%; 8/52). The HCV infection was significantly higher in the older population (>55 years) and exposure to injection from traditional practitioners was identified as a significant () risk factor of infection. Further studies to determine the factors causing the high prevalence of HCV in Rwanda are recommended. Esperance Umumararungu, Fabien Ntaganda, John Kagira, and Naomi Maina Copyright © 2017 Esperance Umumararungu et al. All rights reserved. Utility of Real-Time Quantitative Polymerase Chain Reaction in Detecting Mycobacterium tuberculosis Wed, 11 Jan 2017 00:00:00 +0000 This study aimed to assess the value of real-time quantitative polymerase chain reaction (RT-qPCR) for the detection of Mycobacterium tuberculosis (MTB). Samples from 192 patients with suspected MTB were examined by RT-qPCR and an improved Löwenstein–Jensen (L-J) culture method. To evaluate the diagnostic usefulness of RT-qPCR in detecting MTB, a receiver operating characteristic (ROC) curve for RT-qPCR was generated, and the area under the curve (AUC) as well as a cutoff value was calculated. Using the L-J culture method as the gold standard, accuracy of the RT-qPCR method for detecting MTB was 92.7%, with sensitivity and specificity of 62.5% and 97.02%, respectively. In comparison with the improved L-J culture method, the AUC of RT-qPCR ROC curve was 0.957, which was statistically significant (). The Youden Index reached the maximum value (0.88) for gene copy number of 794.5 IU/mL, which was used as the cutoff value. RT-qPCR detection of MTB yielded results consistent with those of the improved L-J culture method, with high accuracy. RT-qPCR may be used as an auxiliary method for etiological diagnosis of tuberculosis. Zhongquan Lv, Mingxin Zhang, Hui Zhang, and Xinxin Lu Copyright © 2017 Zhongquan Lv et al. All rights reserved. Genetic Structure and Drug Susceptibility Patterns of Mycobacterium tuberculosis Complex Strains Responsible of Human Pulmonary Tuberculosis in the Major Rearing Region in Cameroon Thu, 29 Dec 2016 09:23:22 +0000 Background. Cameroon this last decade continues to present a low contribution of M. africanum and M. bovis in human tuberculosis (TB), while M. bovis was prevalent in cattle but all these pieces of information only concerned West and Center regions. Methods. We carried out the first study in Adamaoua, one of the most rearing regions of Cameroon, on the genetic structure and drug susceptibility of the MTBC strains isolated from newly diagnosed sputum smear-positive patients aged 15 years and above. For that purpose, spoligotyping, a modified 15 standard MIRU/VNTR loci typing, and the proportion method were used. Results. Four hundred and thirty-seven MTBC isolates were analyzed by spoligotyping. Of these, 423 were identified as M. tuberculosis, within the Cameroon family being dominant with 278 (65.7%) isolates; twelve (2.75%) isolates were classified as M. africanum and two as M. bovis. MIRU/VNTR typing of the most prevalent sublineage (SIT 61) suggested that this lineage is not a unique clone as thought earlier but could constitute a group of strains implicated to different pocket of TB transmission. Only M. tuberculosis sublineages were associated with antituberculosis drug resistance. Conclusion. These results showed the weak contribution of M. africanum and M. bovis to human active pulmonary tuberculosis in Cameroon even in the rearing region. Francioli Koro Koro, Alphonse Um Boock, Afi Leslie Kaiyven, Juergen Noeske, Cristina Gutierrez, Christopher Kuaban, François-Xavier Etoa, and Sara Irène Eyangoh Copyright © 2016 Francioli Koro Koro et al. All rights reserved. Quality Improvement Interventions for Early HIV Infant Diagnosis in Northeastern Uganda Mon, 26 Dec 2016 08:33:20 +0000 Introduction. Early infant diagnosis (EID) of human immunodeficiency virus (HIV) ensures prompt treatment and infant survival. In Kaabong Hospital, 20% of HIV exposed infants (HEIs) had access to HIV diagnosis by eight weeks. We aimed to improve EID of HIV by deoxyribonucleic acid-polymerase chain reaction (DNA-PCR) testing by eight weeks from 20 to 100% between June 2014 and November 2015. Method. In this quality improvement (QI) project, EID data was reviewed, gaps prioritized using theme matrix selection, root causes analyzed using fishbone tool, and improvement changes were selected using counter measures matrix but implemented using Plan-Do-Study-Act cycle. Root causes of low first DNA-PCR testing included maternal EID ignorance, absent lost mother-baby pairs (LMBP) tracking system, and no EID performance reviews. Health education, Continuous Medical Education (CMEs), and integration of laboratory and EID services were initial improvement changes used. Results. DNA-PCR testing increased from 20 to 100% between June 2014 and July 2015 and was sustained at 100% until February 2016. Two declines, 67% in September 2014 and 75% in June 2015, due to LMBP were addressed using expert clients and peer mothers, respectively. Conclusion. Formation of WIT, laboratory service integration at MBCP, and task shifting along EID cascade improved EID outcomes at 6 weeks. Jonathan Izudi, Agnes Akot, Grace Paul Kisitu, Pauline Amuge, and Adeodata Kekitiinwa Copyright © 2016 Jonathan Izudi et al. All rights reserved. Evaluation of Two Methods for Determination of CD64 as a Diagnostic Marker of Infection in Critically Ill Adults Tue, 20 Dec 2016 08:17:42 +0000 Objectives. Diagnostic markers of infection have had little innovation over the last few decades. CD64, a marker expressed on the surface of neutrophils, may have utility for this purpose. Methods. This study was conducted in an adult intensive care unit (ICU) in São Paulo, Brazil, with 89 patients. We evaluated CD64 in patients with documented or clinically diagnosed infection (infection group) and controls (patients without any evidence of infection) by two different methodologies: method #1, an in house assay, and method #2, the commercial kit Leuko64 (Trillium Diagnostics). Results. CD64 displayed good discriminating power with a 91.2% sensitivity (95% CI 90.7–91.6%) for detecting infection. The commercial kit (Leuko64) demonstrated higher specificity (87.3%) compared with method #1 as well as better accuracy (88.8%). Conclusions. CD64 seems to be a promising marker of infection in the intensive care setting, with Leuko64 showing a slight advantage. Thiago Zinsly Sampaio Camargo, Alexandre R. Marra, Nydia Strachman Bacal, Eduardo Casaroto, Lilian Moreira Pinto, Jacyr Pasternak, Elivane da Silva Victor, Oscar Fernando Pavão dos Santos, and Michael B. Edmond Copyright © 2016 Thiago Zinsly Sampaio Camargo et al. All rights reserved. Factors Associated with Missed Detection of Mycobacterium tuberculosis by Automated BACTEC MGIT 960 System Mon, 19 Dec 2016 14:10:13 +0000 Despite the demonstration of excellent performance, mycobacterial growth in BACTEC MGIT 960 can go undetected. The aim of this study was to investigate the prevalence of “false-negative” culture sample in Beijing and the potential factors associated with the detection failures by MGIT 960. Of the 577 sputum samples tested, 141 (24.4%) were culture-positive for mycobacteria, of which 133 (94.3%) were automatically determined by MGIT 960 system and 8 (5.7%) were positive for visual growth (false negative by MGIT). Statistical analysis showed that positive grade of specimen had no influence on the false-negative rate by MGIT 960 system (, ). In addition, the mean time to detection (TTD) was 241.4 (range: 224–261) hours for false-negative group and 186.8 (range: 173–199) hours for positive group. The difference in TTD between false-negative and positive groups was statistically significant (). In conclusion, our data demonstrate that the automatic MGIT missed a small portion of bacteriological mycobacterial patients. In addition, the poor growth rate rather than the low grade of AFB smear is associated with the detection failure by MGIT. Our findings highlight the notion that manual inspection for all instrument-negative MGIT tubes will bring about considerable benefit to patients and clinicians. Yu Pang, Biyi Su, Huiwen Zheng, Zhiguo Zhang, Aijing Ma, Yufeng Wang, and Yanlin Zhao Copyright © 2016 Yu Pang et al. All rights reserved. Prospective Study to Assess Progression of Renal Markers after Interruption of Tenofovir due to Nephrotoxicity Sun, 18 Dec 2016 13:14:40 +0000 Background. Prospective studies about the reversibility of tenofovir disoproxil fumarate- (TDF-) related renal impairment remain scarce. Methods. This is an observational prospective study including all patients that presented at our HIV Unit who interrupted TDF owing to nephrotoxicity. We assessed the evolution of renal parameters after discontinuation of this drug. Results. We included 59 patients, who were followed up for 72 weeks. Most were male (41, 69.5%), median (IQR) age was 53 (44; 58) years, and median time receiving TDF-containing regimens was 55.4 (28; 87.7) months. Most patients were receiving PI-based treatments (67%). At the final visit, most of the subjects showed complete recovery (35, 59.3%) or improvement (13 subjects, 22%). Significant improvements were observed in creatinine levels (from 84.9 [73.8; 97.5] to 78 [69.6; 91] μmol/L, ), estimated glomerular filtration rate (eGFR, CKD EPI equation, from 87.7 [67; 99] to 89.9 [73.6; 99.3] mL/min/1.73 m2, ), and number of patients with eGFR <60 mL/min/1.73 m2 (from 9 [15.3%] to 1 [1.7%], ). A trend toward significance was observed in abnormal urine proteinuria/creatinine ratio (from 22 [37%] to 8 [13.6%], ). Conclusions. Our results corroborate the high frequency of complete or partial renal recovery in patients receiving TDF-containing regimens who discontinued therapy owing to nephrotoxicity. Anna Bonjoch, Patricia Echeverría, Núria Perez-Alvarez, Jordi Puig, Carla Estany, Bonaventura Clotet, and Eugènia Negredo Copyright © 2016 Anna Bonjoch et al. All rights reserved. Agreement among Four Prevalence Metrics for Urogenital Schistosomiasis in the Eastern Region of Ghana Sun, 18 Dec 2016 12:22:19 +0000 Few studies assess agreement among Schistosoma haematobium eggs, measured hematuria, and self-reported metrics. We assessed agreement among four metrics at a single time point and analyzed the stability of infection across two time points with a single metric. We used data from the Eastern Region of Ghana and constructed logistic regression models. Girls reporting macrohematuria were 4.1 times more likely to have measured hematuria than girls not reporting macrohematuria (CI95%: 2.1–7.9); girls who swim were 3.6 times more likely to have measured hematuria than nonswimmers (CI95%: 1.6–7.9). For boys, neither self-reported metric was predictive. Girls with measured hematuria in 2010 were 3.3 times more likely to be positive in 2012 (CI95%: 1.01–10.5), but boys showed no association. Boys with measured hematuria in 2008 were 6.0 times more likely to have measured hematuria in 2009 (CI95%: 1.5–23.9) and those with eggs in urine in 2008 were 4.8 times more likely to have eggs in urine in 2009 (CI95%: 1.2–18.8). For girls, measured hematuria in 2008 predicted a positive test in 2009 (OR = 2.8; CI95%: 1.1–6.8), but egg status did not. Agreement between dipstick results and eggs suggests continued dipstick used is appropriate. Self-reported swimming should be further examined. For effective disease monitoring, we recommend annual dipstick testing. Karen Claire Kosinski, Alexandra V. Kulinkina, David Tybor, Dickson Osabutey, Kwabena M. Bosompem, and Elena N. Naumova Copyright © 2016 Karen Claire Kosinski et al. All rights reserved. Tigecycline Therapy for Nosocomial Pneumonia due to Carbapenem-Resistant Gram-Negative Bacteria in Critically Ill Patients Who Received Inappropriate Initial Antibiotic Treatment: A Retrospective Case Study Sun, 04 Dec 2016 11:02:28 +0000 Background. Nosocomial pneumonia due to carbapenem-resistant Gram-negative bacteria (CRGNB) is a growing concern because treatment options are limited and the mortality rate is high. The effect of tigecycline (TGC) on nosocomial pneumonia due to CRGNB in patients who have received inappropriate initial empiric antibiotic treatment (IIAT) is unclear. Therefore, this study aimed to examine the effect of TGC on nosocomial pneumonia due to CRGNB in critically ill patients who had received IIAT. Methods. A retrospective study was conducted in an adult respiratory intensive care unit. Data were obtained and analyzed for all patients who were treated with TGC ≥ 3 days for microbiologically confirmed nosocomial pneumonia due to CRGNB and had experienced initial antibiotic failure. Clinical and microbiological outcomes were investigated. Results. Thirty-one patients with hospital-acquired pneumonia or ventilator-associated pneumonia were included in the study. The majority of the responsible organisms were carbapenem-resistant Acinetobacter baumannii (67.7%), followed by Klebsiella pneumoniae (16.1%) and Escherichia coli (9.7%). Twenty patients were treated with high-dose TGC therapy (100 mg every 12 h after a 200 mg loading dose), and the others received a standard-dose therapy (50 mg every 12 h after a 100 mg loading dose). The duration of TGC therapy was days. The global clinical cure rate and the microbiological eradication rate were 48.4% and 61.3%, respectively. The overall ICU mortality rate was 45.2%. A higher score on the Acute Physiology and Chronic Health Evaluation II and a longer duration of IIAT were associated with clinical failure. High-dose TGC therapy had a higher clinical success rate [65.0% (13/20) versus 18.2% (2/11), ] and a lower ICU mortality rate [30.0% (6/20) versus 72.7% (8/11), ] than the standard-dose therapy. Conclusions. TGC, especially a high-dose regimen, might be a justifiable option for critically ill patients with nosocomial pneumonia due to CRGNB who have received IIAT when the options for these patients are limited. Xiaomai Wu, Yefei Zhu, Qiuying Chen, Liuyang Gong, Jian Lin, Dongqing Lv, and Jiaxi Feng Copyright © 2016 Xiaomai Wu et al. All rights reserved. Spinal Epidural Abscess: A Review with Special Emphasis on Earlier Diagnosis Thu, 01 Dec 2016 09:20:11 +0000 Spinal epidural abscess (SEA) is an uncommon but serious condition with significant morbidity and mortality. The prognosis of SEA is highly dependent on the timeliness of its diagnosis before neurological deficits develop. Unfortunately, often due to its nonspecific presentation, such as back pain, the diagnosis of SEA may be delayed in up to 75% of cases. Although many risk factors for SEA can be found in the published literature, their utility is limited by their frequent lack of objective evidence, numerousness, and absence in a significant proportion of cases. In this review, we call for a more discriminate evidence-based use of the term “risk factor” when discussing SEA and explore several approaches to its earlier diagnosis, including a simple algorithm based on its pathophysiology and serum C-reactive protein or erythrocyte sedimentation rate. Allison Bond and Farrin A. Manian Copyright © 2016 Allison Bond and Farrin A. Manian. All rights reserved. A Quantitative Real-Time RT-PCR Assay for the Detection of Venezuelan equine encephalitis virus Utilizing a Universal Alphavirus Control RNA Tue, 29 Nov 2016 13:08:46 +0000 Venezuelan equine encephalitis virus (VEEV) is an Alphavirus from the family Togaviridae that causes epizootic outbreaks in equids and humans in Central and South America. So far, most studies use conventional reverse transcriptase PCR assays for the detection of the different VEEV subtypes. Here we describe the development of a TaqMan quantitative real-time reverse transcriptase PCR assay for the specific detection and quantitation of all VEEV subtypes which uses in parallel a universal equine encephalitis virus control RNA carrying target sequences of the three equine encephalitis viruses. The control RNA was used to generate standard curves for the calculation of copy numbers of viral genome of Eastern equine encephalitis virus (EEEV), Western equine encephalitis virus (WEEV), and VEEV. The new assay provides a reliable high-throughput method for the detection and quantitation of VEEV RNA in clinical and field samples and allows a rapid differentiation from potentially cocirculating EEEV and WEEV strains. The capability to detect all known VEEV variants was experimentally demonstrated and makes this assay suitable especially for the surveillance of VEEV. Ariel Vina-Rodriguez, Martin Eiden, Markus Keller, Winfried Hinrichs, and Martin H. Groschup Copyright © 2016 Ariel Vina-Rodriguez et al. All rights reserved. Ebola Virus Infection among Western Healthcare Workers Unable to Recall the Transmission Route Sun, 27 Nov 2016 13:13:13 +0000 Introduction. During the 2014–2016 West-African Ebola virus disease (EVD) outbreak, some HCWs from Western countries became infected despite proper equipment and training on EVD infection prevention and control (IPC) standards. Despite their high awareness toward EVD, some of them could not recall the transmission routes. We explored these incidents by recalling the stories of infected Western HCWs who had no known directly exposures to blood/bodily fluids from EVD patients. Methodology. We carried out conventional and unconventional literature searches through the web using the keyword “Ebola” looking for interviews and reports released by the infected HCWs and/or the healthcare organizations. Results. We identified fourteen HCWs, some infected outside West Africa and some even classified at low EVD risk. None of them recalled accidents, unintentional exposures, or any IPC violation. Infection transmission was thus inexplicable through the acknowledged transmission routes. Conclusions. We formulated two hypotheses: inapparent exposures to blood/bodily fluids or transmission due to asymptomatic/mildly symptomatic carriers. This study is in no way intended to be critical with the healthcare organizations which, thanks to their interventions, put an end to a large EVD outbreak that threatened the regional and world populations. Stefano Petti, Carmela Protano, Giuseppe Alessio Messano, and Crispian Scully Copyright © 2016 Stefano Petti et al. All rights reserved. Clinical and Microbiological Characteristics of Visceral Leishmaniasis Outbreak in a Northern Italian Nonendemic Area: A Retrospective Observational Study Wed, 23 Nov 2016 14:03:58 +0000 Background. Visceral leishmaniasis (VL) caused by Leishmania infantum is endemic in the Mediterranean area. In the last decades a northward spread of the parasite has been observed in Italy. This paper describes a VL outbreak in Modena province (Emilia-Romagna, Northern Italy) between 2012 and 2015. Methods. Retrospective, observational study to evaluate epidemiological, microbiological characteristics, and clinical management of VL in patients referring to Policlinico Modena Hospital. Results. Sixteen cases of VL occurred in the study period. An immunosuppressive condition was present in 81.3%. Clinical presentation included anemia, fever, leukopenia, thrombocytopenia, and hepatosplenomegaly. Serology was positive in 73.3% of cases, peripheral blood PCR in 92.3%, and bone marrow blood PCR in 100%. Culture was positive in 3/6 cases (50%) and all the isolates were identified as L. infantum by ITS1/ITS2 sequencing. The median time between symptom onset and diagnosis was 22 days (range 6–131 days). All patients were treated with liposomal amphotericin b. 18.8% had a VL recurrence and were treated with miltefosine. Attributable mortality was 6.3%. Conclusions. VL due to L. infantum could determine periodical outbreaks, as the one described; thus it is important to include VL in the differential diagnosis of fever of unknown origin, even in low-endemic areas. E. Franceschini, C. Puzzolante, M. Menozzi, L. Rossi, A. Bedini, G. Orlando, W. Gennari, M. Meacci, G. Rugna, E. Carra, M. Codeluppi, and C. Mussini Copyright © 2016 E. Franceschini et al. All rights reserved. HBV Infection Status and the Risk of Cholangiocarcinoma in Asia: A Meta-Analysis Wed, 23 Nov 2016 09:42:31 +0000 Background. The inconsistent finding was between hepatitis B virus (HBV) infections and cholangiocarcinoma (CCA). This meta-analysis is to explore this relationship in Asia. Methods. A literature search was performed using PubMed, Web of Science, and Cochrane Library to October 30, 2015. Pooled incidence rate and OR with 95% CI were calculated using STATA 11.0. Results. Thirty-nine studies were included. The pooled incidence rate of CCA patients with HBV infection was 31% (95% CI 22%–39%). The pooled OR showed increased risk of CCA incidence with HBV infection (OR = 2.72, 95% CI 1.90–3.88), especially in ICC (OR = 3.184, 95% CI 2.356–4.302), while it showed no risk in ECC (OR = 1.407, 95% CI 0.925–2.141). Also, the pooled OR showed increased risk of ICC and ECC incidence (OR = 6.857, 95% CI 4.421–10.633 and OR = 1.740, 95% CI 1.260–2.404) in patients with HBsAg+/HBcAb+. The pooled OR showed increased risk of ICC incidence (OR = 1.410, 95% CI 1.095–1.816) in patients with HBsAg−/HBcAb+. Conclusion. It is suggested that HBV infection is associated with an increased risk of CCA in Asia. Two HBV infection models (HBsAg+/HBcAb+ and HBsAg−/HBcAb+) increase the risk of CCA, and patients with HBsAg−/HBcAb+ also had a risk of ICC. This trial is registered with PROSPERO CRD42015029264. Hao Zhang, Biqing Zhu, He Zhang, Jianxin Liang, and Wenting Zeng Copyright © 2016 Hao Zhang et al. All rights reserved. A First Insight into the Genetic Diversity and Drug Susceptibility Pattern of Mycobacterium tuberculosis Complex in Zhejiang, China Tue, 22 Nov 2016 07:48:38 +0000 In this study, our aim was to determine the predominant genotypes among the Mycobacterium tuberculosis (MTB) strains circulating in Zhejiang Province. In addition, we also sought to determine the potential associations between MTB genotypes and susceptibility to first-line drugs. Out of these isolates, 673 (71.6%) were classified into the Beijing genotype, while the other 267 (28.4%) were from non-Beijing families. The highest proportion of Beijing genotype was found in Huzhou (80.0%) and the lowest in Lishui (48.3%). Statistical analysis revealed that there was a significant difference in the prevalence of Beijing genotype among different regions (χ2 = 17.57, P = 0.04). In addition, the overall proportions of drug resistance to INH, RIF, SM, and EMB were 13.2% (124/940), 21.8% (75/940), 3.4% (32/940), and 5.9% (55/940) in Zhejiang, respectively. Further comparison revealed that there was no significant difference in drug susceptibility profiles between Beijing and non-Beijing strains (P > 0.05). In conclusion, we describe the genetic diversity and drug susceptibility pattern of MTB in Zhejiang for the first time. Our data demonstrate that Beijing genotype is the predominant lineage in Zhejiang, while the distribution of Beijing-genotype strains shows geographic diversity. In addition, no correlation is observed between Beijing genotype and anti-TB drug resistance. Zhengwei Liu, Yu Pang, Songhua Chen, Beibei Wu, Haibo He, Aizhen Pan, and Xiaomeng Wang Copyright © 2016 Zhengwei Liu et al. All rights reserved. Comparative Evaluation of Anthelmintic Activity of Edible and Ornamental Pomegranate Ethanolic Extracts against Schistosoma mansoni Mon, 21 Nov 2016 06:41:36 +0000 Due to the development of praziquantel (PZQ) schistosomes resistant strains, the discovery of new antischistosomal agents is of high priority in research. This work reported the in vitro and in vivo effects of the edible and ornamental pomegranate extracts against Schistosoma mansoni. Leaves and stem bark ethanolic extracts of both dried pomegranates were prepared at 100, 300, and 500 μg/mL for in vitro and 600 and 800 mg/kg for in vivo. Adult worms Schistosoma mansoni in RPMI-1640 medium for in vitro and S. mansoni infected mice for in vivo tests were obtained from Theodor Bilharz Research Institute, Cairo, Egypt. In vitro activity was manifested by significant coupled worms separation, reduction of motor activity, lethality, and ultrastructural tegumental alterations in adult worms. In vivo activity was manifested revealed by significant reduction of hepatic granulomas number and diameter, decreased number of bilharzial eggs in liver tissues, lowered liver inflammatory infiltration, decreased hepatic fibrosis, and inducible nitric oxide synthase (iNOS) expression. Ethanolic stem bark extract of edible pomegranate exhibited highest antischistosomal activities both in vitro and in vivo. Therefore, pomegranate showed a good potential to be used as a promising new candidate for the development of new schistosomicidal agents. Doaa A. Yones, Dalia M. Badary, Hesham M. B. Sayed, Soad A. H. Bayoumi, Azza A. Khalifa, and Ahmed M. El-Moghazy Copyright © 2016 Doaa A. Yones et al. All rights reserved. Low-Intensity Ultrasound Combined with Hematoporphyrin Monomethyl Ether in the Treatment of Experimental Periodontitis in Rats Wed, 16 Nov 2016 12:33:31 +0000 Objectives. This study aims to evaluate the efficacy of hematoporphyrin monomethyl ether- (HMME-) mediated sonodynamic therapy (SDT) on experimental periodontal disease in rats. Methods. Periodontal disease was induced by submerging ligatures at the first maxillary molar subgingival region in forty-eight male SD rats. After 30 days, the ligatures were removed. The rats were randomly allocated into four groups; the experimental SDT group was treated through hypodermic injection of 40 μg/mL HMME and 3 W/cm2 low-intensity ultrasound irradiation (1 MHz, 600 s). Those in control groups received 40 μg/mL HMME alone (control 1 group) or 3 W/cm2 ultrasound irradiation alone (control 2 group) or were subjected to neither HMME nor ultrasound (control 3 group). After 10 days of treatment, all rats were euthanized, the maxilla was obtained for histological examination, and the alveolar bone level was evaluated by histometric analysis. Results. The control groups showed more bone loss () after 10 days of treatment than the SDT group. There is no significant difference among the control groups (). Conclusions. HMME mediated SDT was an effective therapy of experimental periodontal tissue in rats. Deshu Zhuang, Zongshan Ji, Liangjia Bi, Xiaochun Wang, Qi Zhou, and Wenwu Cao Copyright © 2016 Deshu Zhuang et al. All rights reserved. A De-O-acylated Lipooligosaccharide-Based Adjuvant System Promotes Antibody and Th1-Type Immune Responses to H1N1 Pandemic Influenza Vaccine in Mice Mon, 07 Nov 2016 08:06:13 +0000 Vaccine adjuvants are agents that are used to promote immune responses to vaccine antigens and thereby to enhance the protective efficacy of the vaccines. In this study, we investigated the adjuvant activity of CIA06, an adjuvant system that is composed of a toll-like receptor 4 agonist de-O-acylated lipooligosaccharide (dLOS) and aluminum hydroxide, on the H1N1 pandemic influenza vaccine Greenflu-S® in mice. CIA06 significantly enhanced influenza-specific serum IgG, hemagglutination-inhibition, and virus-neutralizing antibody titers, which eliminated vaccine dose-dependency in the antibody response. Mice immunized with the CIA06-adjuvanted Greenflu-S showed Th1-type-predominant cytokine profiles, and both CD4+ and CD8+ T cell responses were induced. Immunization of mice with the CIA06-adjuvanted vaccine reduced the mortality and morbidity of mice upon lethal challenges with influenza virus, and no excessive inflammatory responses were observed in the lung tissues of the immunized mice after viral infection. These data suggest that the dLOS-based adjuvant system CIA06 can be used to promote the immune responses to influenza vaccine or to spare antigen dose without causing harmful inflammatory responses. Ji In Ryu, Shin Ae Park, Seo Ri Wui, Ara Ko, Ji Eun Han, Jung Ah Choi, Man Ki Song, Kwang Sung Kim, Yang Je Cho, and Na Gyong Lee Copyright © 2016 Ji In Ryu et al. All rights reserved. Modelling Seasonal Brucellosis Epidemics in Bayingolin Mongol Autonomous Prefecture of Xinjiang, China, 2010–2014 Sun, 30 Oct 2016 12:40:23 +0000 Brucellosis is one of the severe public health problems; the cumulative number of new human brucellosis cases reached 211515 from 2010 to 2014 in China. Bayingolin Mongol Autonomous Prefecture is situated in the southeast of Xinjiang, where brucellosis infection occurs every year. Based on the reported data of newly acute human brucellosis cases for each season in Bayingolin Mongol Autonomous Prefecture, we proposed a susceptible, exposed, infected, and vaccinated (SEIV) model with periodic transmission rates to investigate the seasonal brucellosis transmission dynamics among sheep/cattle and from sheep/cattle to humans. Compared with the criteria of MAPE and RMSPE, the model simulations agree to the data on newly acute human brucellosis. We predict that the number of newly acute human brucellosis is increasing and will peak 15325 [95% CI: 11920–18242] around the summer of 2023. We also estimate the basic reproduction number [95% CI: 2.5129–2.6225] and perform some sensitivity analysis of the newly acute human brucellosis cases and the basic reproduction number in terms of model parameters. Our study demonstrates that reducing the birth number of sheep/cattle, raising the slaughter rate of infected sheep/cattle, increasing the vaccination rate of susceptible sheep/cattle, and decreasing the loss rate of vaccination are effective strategies to control brucellosis epidemic. Pengwei Lou, Lei Wang, Xueliang Zhang, Jiabo Xu, and Kai Wang Copyright © 2016 Pengwei Lou et al. All rights reserved. Systematic Identification and Bioinformatic Analysis of MicroRNAs in Response to Infections of Coxsackievirus A16 and Enterovirus 71 Mon, 24 Oct 2016 14:18:58 +0000 Hand, foot, and mouth disease (HFMD), mainly caused by coxsackievirus A16 (CVA16) and enterovirus 71 (EV71) infections, remains a serious public health issue with thousands of newly diagnostic cases each year since 2008 in China. The mechanisms underlying viral infection, however, are elusive to date. In the present study, we systematically investigated the host cellular microRNA (miRNA) expression patterns in response to CVA16 and EV71 infections. Through microarray examination, 27 miRNAs (15 upregulated and 12 downregulated) were found to be coassociated with the replication process of two viruses, while the expression levels of 15 and 5 miRNAs were significantly changed in CVA16- and EV71-infected cells, respectively. A great number of target genes of 27 common differentially expressed miRNAs were predicted by combined use of two computational target prediction algorithms, TargetScan and MiRanda. Comprehensive bioinformatic analysis of target genes in GO categories and KEGG pathways indicated the involvement of diverse biological functions and signaling pathways during viral infection. These results provide an overview of the roles of miRNAs in virus-host interaction, which will contribute to further understanding of HFMD pathological mechanisms. Zheng Zhu, Yuhua Qi, Huan Fan, Lunbiao Cui, and Zhiyang Shi Copyright © 2016 Zheng Zhu et al. All rights reserved. Efficacy of Scutellaria baicalensis for the Treatment of Hand, Foot, and Mouth Disease Associated with Encephalitis in Patients Infected with EV71: A Multicenter, Retrospective Analysis Thu, 20 Oct 2016 14:10:05 +0000 This study aimed to evaluate the clinical efficacy and safety of using the traditional Chinese herbal medicine Scutellaria baicalensis for the treatment of severe HFMD in 725 patients aged >1 year in a multicenter, retrospective analysis. The patients were divided into the S. baicalensis and ribavirin groups, and the temperatures, presence or absence of skin rashes and oral lesions, nervous system (NS) involvement, and viral loads of the patients, as well as the safety of the treatments, were evaluated. The median duration of fever, median time to NS involvement, and the number of patients with oral ulcers and/or vesicles, as well as skin rashes, were decreased in the S. baicalensis group compared with the ribavirin group. In addition, the EV71 viral loads were decreased in the S. baicalensis group, suggesting that S. baicalensis exerted more potent antiviral effects compared with ribavirin. The present study demonstrated that S. baicalensis was suitable for the treatment of severe HFMD in patients aged >1 year, since it was shown to rapidly relieve fever, attenuate oral lesions and rashes, and improve NS involvement. Furthermore, it was demonstrated to be relatively safe for topical application. Hailong Lin, Jian Zhou, Kaichun Lin, Hongjiao Wang, Zunhong Liang, Xingshuai Ren, Leting Huang, and Chan Xia Copyright © 2016 Hailong Lin et al. All rights reserved. NRAMP1 Polymorphisms like Susceptibility Marker in Mexican Focus of Cutaneous Leishmaniasis Tue, 18 Oct 2016 13:04:16 +0000 Cutaneous leishmaniasis (CL) is endemic in Campeche state, Mexico. Host and parasite factors are involved in the establishment and development of CL. Host factors include immune response and genetic background. NRAMP1 (Natural Resistance Associated Macrophage Protein 1) is important in innate immunity. Polymorphisms in NRAMP1 have been associated with susceptibility or resistance to infectious and autoimmune diseases. To study the association of NRAMP1 mutations with CL in patients from Calakmul, Campeche, samples from 115 CL patients and 69 samples of healthy people from the same area were evaluated. Five regions in NRAMP1 were amplified and digested, looking for mutations in the promoter region (−524G/C), exon 3 (274C/T), exon 8 (823 C7T), and exon 15 (G/A) and deletion of 4 bp in the 3′UTR region. We found a statistical association between polymorphisms in 3′UTR region and exon 8 and CL [; ; OR = 17.00; IC of 95% (2.24–128.99)]. Some patients who needed more than 40 doses of Glucantime to heal injuries presented mutations in exons 3, 8, and 15. Multiple or ear lesions were not associated with NRAMP1 polymorphism. Mirsha Pamela Hernández-Rivera, Alicia Ramírez-Ramírez, Adelaido Chiñas-Pérez, Amalia Monroy-Ostria, Mario Eugenio Cancino-Díaz, and Omar Hernández-Montes Copyright © 2016 Mirsha Pamela Hernández-Rivera et al. All rights reserved. First Observation of Leptospira interrogans in the Lungs of Rattus norvegicus Wed, 05 Oct 2016 15:00:08 +0000 We report the first two cases of pulmonary presence of leptospires in apparently healthy rats captured in a city park in Lyon (France). Only renal carriage of Leptospira has been described in the literature. Blood serology was performed in parallel with molecular and histological analyses of the kidney and lung samples. We isolated leptospires from the kidneys of two out of three seropositive wild rats. These results were confirmed by specific detection of pathogenic Leptospira by real-time PCR. Moreover, Leptospira DNA was detected in lung tissues. Immunohistochemistry and Warthin-Starry staining revealed that leptospires were present on the surface of the ciliated epithelium of the bronchi. Using PCR of the rrs (16S) gene and Multispacer Sequence Typing, DNA extracts of the kidney and lung were identified as belonging to Leptospira interrogans serovar Icterohaemorrhagiae “CHU Réunion.” This first observation of the presence Leptospira in the lung with simultaneous renal carriage will require further study in future on several target organs to gain a better understanding of the Leptospira infection in wild rat. Anne-Laure Zilber, Patrick Belli, Marc Artois, Angeli Kodjo, and Zoheira Djelouadji Copyright © 2016 Anne-Laure Zilber et al. All rights reserved. Epidemiology of American Tegumentary Leishmaniasis and Trypanosoma cruzi Infection in the Northwestern Argentina Thu, 29 Sep 2016 13:42:44 +0000 Background. Endemic areas of tegumentary leishmaniasis (TL) in Salta, Argentina, present some overlap zones with the geographical distribution of Chagas disease, with mixed infection cases being often detected. Objectives. The purpose of this study was to determine the magnitude of Leishmania sp. infection and potential associated risk factors, the serologic prevalence of T. cruzi, and the presence of T. cruzi-Leishmania sp. mixed infection in a region of the northwest of Argentina. Methods. Cross-sectional studies were conducted to detect TL prevalence and T. cruzi seroprevalence. A case-control study was conducted to examine leishmaniasis risk factors. Results. Prevalence of TL was 0.17%, seroprevalence of T. cruzi infection was 9.73%, and mixed infection proportion—within the leishmaniasic patients group—was 16.67%. The risk factors associated with TL transmission were sex, age, exposure to bites at work, staying outdoors more than 10 hours/day, bathing in the river, and living with people who had lesions or were infected during the study. Discussion. The endemic pattern of TL seems to involve exposure of patients to vectors in wild as well as peridomestic environment. Cases of T. cruzi infection are apparently due to migration. Therefore, a careful epidemiological surveillance is necessary due to the contraindication of antimonial administration to chagasic patients. Carlos L. Hoyos, Silvana P. Cajal, Marisa Juarez, Jorge D. Marco, Anahí M. Alberti D’Amato, Melina Cayo, Irma Torrejón, Rubén O. Cimino, Patricio Diosque, Alejandro J. Krolewiecki, Julio R. Nasser, and José F. Gil Copyright © 2016 Carlos L. Hoyos et al. All rights reserved. Differences in Rhodococcus equi Infections Based on Immune Status and Antibiotic Susceptibility of Clinical Isolates in a Case Series of 12 Patients and Cases in the Literature Wed, 10 Aug 2016 11:39:06 +0000 Rhodococcus equi is an unusual zoonotic pathogen that can cause life-threatening diseases in susceptible hosts. Twelve patients with R. equi infection in Kentucky were compared to 137 cases reported in the literature. Although lungs were the primary sites of infection in immunocompromised patients, extrapulmonary involvement only was more common in immunocompetent patients (). Mortality in R. equi-infected HIV patients was lower in the HAART era (8%) than in pre-HAART era (56%) (), suggesting that HAART improves prognosis in these patients. Most (85–100%) of clinical isolates were susceptible to vancomycin, clarithromycin, rifampin, aminoglycosides, ciprofloxacin, and imipenem. Interestingly, there was a marked difference in susceptibility of the isolates to cotrimoxazole between Europe (35/76) and the US (15/15) (). Empiric treatment of R. equi infection should include a combination of two antibiotics, preferably selected from vancomycin, imipenem, clarithromycin/azithromycin, ciprofloxacin, rifampin, or cotrimoxazole. Local antibiograms should be checked prior to using cotrimoxazole due to developing resistance. Praveen Gundelly, Yasuhiro Suzuki, Julie A. Ribes, and Alice Thornton Copyright © 2016 Praveen Gundelly et al. All rights reserved. Health Care Providers’ Knowledge and Practice Gap towards Joint Zoonotic Disease Surveillance System: Challenges and Opportunities, Gomma District, Southwest Ethiopia Sun, 07 Aug 2016 13:47:20 +0000 Background. Health care providers play a crucial role for realization of joint zoonotic diseases surveillance by human and animal health sectors, yet there is limited evidence. Hence, this study aimed to determine knowledge and practice gap of health care providers towards the approach for Rabies and Anthrax in Southwest Ethiopia. Methods. A cross-sectional survey was conducted from December 16, 2014, to January 14, 2015. Eligible health care providers were considered for the study. Data were entered in to Epi-data version 3.1 and analyzed using SPSS version 20. Results. A total of 323 (92.02%) health care providers participated in the study. Three hundred sixteen (97.8%) of participants reported that both human and animal health sectors can work together for zoonotic diseases while 96.9% of them replied that both sectors can jointly conduct surveillance. One hundred seventeen (36.2%) of them reported that their respective sectors had conducted joint surveillance for zoonotic diseases. Their involvement was, however, limited to joint outbreak response. Conclusion. There is good opportunity in health care providers’ knowledge even though the practice was unacceptably low and did not address all surveillance components. Therefore, formal joint surveillance structure should be in place for optimal implementation of surveillance. Desta Hiko Gemeda, Abiot Girma Sime, Kifle Woldemichael Hajito, Benti Deresa Gelalacha, Wubit Tafese, and Tsegaye Tewelde Gebrehiwot Copyright © 2016 Desta Hiko Gemeda et al. All rights reserved. HMBA Enhances Prostratin-Induced Activation of Latent HIV-1 via Suppressing the Expression of Negative Feedback Regulator A20/TNFAIP3 in NF-κB Signaling Wed, 27 Jul 2016 08:15:07 +0000 In the past decade, much emphasis has been put on the transcriptional activation of HIV-1, which is proposed as a promised strategy for eradicating latent HIV-1 provirus. Two drugs, prostratin and hexamethylene bisacetamide (HMBA), have shown potent effects as inducers for releasing HIV-1 latency when used alone or in combination, although their cellular target(s) are currently not well understood, especially under drug combination. Here, we have shown that HMBA and prostratin synergistically release HIV-1 latency via different mechanisms. While prostratin strongly stimulates HMBA-induced HIV-1 transcription via improved P-TEFb activation, HMBA is capable of boosting NF-κB-dependent transcription initiation by suppressing prostratin-induced expression of the deubiquitinase A20, a negative feedback regulator in the NF-κB signaling pathway. In addition, HMBA was able to increase prostratin-induced phosphorylation and degradation of NF-κB inhibitor IκBα, thereby enhancing and prolonging prostratin-induced nuclear translocation of NF-κB, a prerequisite for stimulation of transcription initiation. Thus, by blocking the negative feedback circuit, HMBA functions as a signaling enhancer of the NF-κB signaling pathway. Duchu Chen, Huiping Wang, Jude Juventus Aweya, Yanheng Chen, Meihua Chen, Xiaomeng Wu, Xiaonan Chen, Jing Lu, Ruichuan Chen, and Min Liu Copyright © 2016 Duchu Chen et al. All rights reserved. Pains and Gains from China’s Experiences with Emerging Epidemics: From SARS to H7N9 Wed, 20 Jul 2016 16:46:30 +0000 Over the recent decades, China experienced several emerging virus outbreaks including those caused by the severe acute respiratory syndrome- (SARS-) coronavirus (Cov), H5N1 virus, and H7N9 virus. The SARS tragedy revealed faults in China’s infectious disease prevention system, propelling the Chinese government to enact reforms that enabled better combating of the subsequent H1N1 and H7N9 avian flu epidemics. The system is buttressed by three fundamental, mutually reinforcing components: (1) enduring government administration reforms, including legislation establishing a unified public health emergency management system; (2) prioritized funding for biotechnology and biomedicine industrialization, especially in the areas of pathogen identification, drug production, and the development of vaccines and diagnostics; and (3) increasing investment for public health and establishment of a rapid-response infectious diseases prevention and control system. China is now using its hard-gained experience to support the fight against Ebola in Africa and the Middle East Respiratory Syndrome in its own country. Pengfei Wei, Zelang Cai, Jinwen Hua, Weijia Yu, Jiajie Chen, Kang Kang, Congling Qiu, Lanlan Ye, Jiayun Hu, and Kunmei Ji Copyright © 2016 Pengfei Wei et al. All rights reserved. Comparison of the Hemagglutination Inhibition Test and IgG ELISA in Categorizing Primary and Secondary Dengue Infections Based on the Plaque Reduction Neutralization Test Thu, 30 Jun 2016 10:36:13 +0000 Secondary dengue infection by heterotypic serotypes is associated with severe manifestations of disease, that is, dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). The World Health Organization (WHO) has recommended criteria based on the hemagglutination inhibition (HI) test to distinguish between primary and secondary dengue infections. Since the HI test has practical limitations and disadvantages, we evaluated the accuracy of WHO HI criteria and compared it with criteria based on an IgG enzyme-linked immunosorbent assay (ELISA) using a plaque reduction neutralization test (PRNT) as the gold standard. Both WHO HI criteria and IgG ELISA criteria performed strongly (16/16) in determining primary infection. However, to determine secondary infection, the IgG ELISA criteria performed better (72/73) compared to the WHO HI criteria (23/73). Nurhayati Lukman, Gustiani Salim, Herman Kosasih, Nugroho Harry Susanto, Ida Parwati, Silvita Fitri, Bachti Alisjahbana, Susana Widjaja, and Maya Williams Copyright © 2016 Nurhayati Lukman et al. All rights reserved. Seroprevalence and Associated Risk Factors for Toxoplasma gondii Infection in Healthy Blood Donors: A Cross-Sectional Study in Sonora, Mexico Thu, 30 Jun 2016 10:32:32 +0000 Toxoplasma gondii (T. gondii) can be transmitted by blood transfusion. We determined the prevalence of T. gondii infection in healthy blood donors in Hermosillo city, Mexico, and the association of infection with T. gondii with the sociodemographic, clinical, and behavioral characteristics of blood donors. Four hundred and eight blood donors who attended two public blood banks in Hermosillo city were examined for anti-T. gondii IgG and IgM antibodies by using enzyme-linked immunoassays. Of the 408 blood donors (mean age ; range 18–60 years old) studied, 55 (13.5%) were positive for anti-T. gondii IgG antibodies, and 12 (21.8%) of them were also positive for anti-T. gondii IgM antibodies. Multivariate analysis showed that seropositivity to T. gondii was associated with age (OR = 1.74; 95% CI: 1.03–2.94; ) and tobacco use (OR = 2.09; 95% CI: 1.02–4.29; ). Seropositivity to T. gondii was correlated with the number of pregnancies, deliveries, and cesarean sections. The seroprevalence of T. gondii infection in blood donors in Sonora is the highest reported in blood donors in northern Mexico so far. This is the first report of an association of T. gondii exposure and tobacco use. Further research to confirm this association is needed. Cosme Alvarado-Esquivel, Antonio Rascón-Careaga, Jesús Hernández-Tinoco, María Alba Guadalupe Corella-Madueño, Luis Francisco Sánchez-Anguiano, María Lourdes Aldana-Madrid, Edgar Velasquez-Vega, Trinidad Quizán-Plata, José Luis Navarro-Henze, Joel Alberto Badell-Luzardo, José María Gastélum-Cano, and Oliver Liesenfeld Copyright © 2016 Cosme Alvarado-Esquivel et al. All rights reserved. Preventive Effects of Houttuynia cordata Extract for Oral Infectious Diseases Mon, 20 Jun 2016 07:41:44 +0000 Houttuynia cordata (HC) (Saururaceae) has been used internally and externally as a traditional medicine and as an herbal tea for healthcare in Japan. Our recent survey showed that HC poultice (HCP) prepared from smothering fresh leaves of HC had been frequently used for the treatment of purulent skin diseases with high effectiveness. Our experimental study also demonstrated that ethanol extract of HCP (eHCP) has antibacterial, antibiofilm, and anti-inflammatory effects against S. aureus which caused purulent skin diseases. In this study, we focused on novel effects of HCP against oral infectious diseases, such as periodontal disease and dental caries. We determined the antimicrobial and antibiofilm effects of water solution of HCP ethanol extract (wHCP) against important oral pathogens and investigated its cytotoxicity and anti-inflammatory effects on human oral epithelial cells. wHCP had moderate antimicrobial effects against some oral microorganisms and profound antibiofilm effects against Fusobacterium nucleatum, Streptococcus mutans, and Candida albicans. In addition, wHCP had no cytotoxic effects and could inhibit interleukin-8 and CCL20 productions by Porphyromonas gingivalis lipopolysaccharide-stimulated human oral keratinocytes. Our findings suggested that wHCP may be clinically useful for preventing oral infectious diseases as a mouthwash for oral care. Yasuko Sekita, Keiji Murakami, Hiromichi Yumoto, Takashi Amoh, Natsumi Fujiwara, Shohei Ogata, Takashi Matsuo, Yoichiro Miyake, and Yoshiki Kashiwada Copyright © 2016 Yasuko Sekita et al. All rights reserved. Epidemiology and Outcome of Ventilator-Associated Pneumonia in a Heterogeneous ICU Population in Qatar Mon, 13 Jun 2016 07:59:42 +0000 Objective. The purpose of this study is to collect data on epidemiology, microbiology, and outcome of VAP in our ICUs for reevaluation of the therapeutic strategies. Methods. This retrospective study involved all adult patients, 15 years of age or older, diagnosed with VAP in multidisciplinary ICUs at Hamad General Hospital between January 2010 and December 2012. Results. A total of 106 patients were enrolled. The mean incidence of VAP was 5.0 per 1000 ventilator-days. It was predominant among younger age group (<60 years), male patients (80.2%), and trauma ICU admissions (49.0%). The most common comorbidity was hypertension (34%) and polytrauma (36.8%) was the most frequent admission diagnosis. 30-day mortality was 23.6% and it was significantly higher in ≥60 years age group, female gender, patients with diabetes mellitus, hypertension, chronic respiratory disease, ≥1 comorbidity, and poor functional status, smokers, medical and surgical ICU admissions, and patients with previous stay in medical/surgical wards, inappropriate empirical therapy, and admission diagnosis of respiratory failure. Gram-negative bacilli were the most frequent respiratory specimen isolates, Pseudomonas spp. being the most common. Majority of our Acinetobacter isolates were multidrug resistant. Conclusion. The incidence of VAP in our ICUs was low. Higher mortality rates were observed in certain subgroup of patients. Resistance to commonly used antimicrobials is likely to require reevaluation of the therapeutic strategies at our institution. Husain Shabbir Ali, Fahmi Yousef Khan, Saibu George, Nissar Shaikh, and Jameela Al-Ajmi Copyright © 2016 Husain Shabbir Ali et al. All rights reserved. Efficacy of Linezolid and Fosfomycin in Catheter-Related Biofilm Infection Caused by Methicillin-Resistant Staphylococcus aureus Sun, 05 Jun 2016 09:24:17 +0000 As long-standing clinical problems, catheter-related infections and other chronic biofilm infections are more difficult to treat due to the high antibiotic resistance of biofilm. Therefore, new treatments are needed for more effective bacteria clearance. In this study, we evaluated the antibacterial activities of several common antibiotics alone and their combinations against biofilm-embedded methicillin-resistant staphylococcus aureus (MRSA) infections, both in vitro and in vivo. In brief, fosfomycin, levofloxacin, and rifampin alone or in combination with linezolid were tested in vitro against planktonic and biofilm-embedded MRSA infection in three MRSA stains. The synergistic effects between linezolid and the other three antibiotics were assessed by fractional inhibitory concentration index (FICI) and time-kill curves, where the combination of linezolid plus fosfomycin showed the best synergistic effect in all strains. For further evaluation in vivo, we applied the combination of linezolid and fosfomycin in a catheter-related biofilm rat model and found that viable bacteria counts in biofilm were significantly reduced after treatment (). In summary, we have shown here that the combination of linezolid and fosfomycin treatment had improved therapeutic effects on biofilm-embedded MRSA infection both in vitro and in vivo, which provided important basis for new clinical therapy development. Dong Chai, Xu Liu, Rui Wang, Yan Bai, and Yun Cai Copyright © 2016 Dong Chai et al. All rights reserved. The Efficacy and Immunomodulatory Effects of Ulinastatin and Thymosin α1 for Sepsis: A Systematic Review and Meta-Analysis Tue, 31 May 2016 09:13:34 +0000 Objective. To systematically review the efficacy and potential immunomodulatory effect of ulinastatin combined with thymosin α1 (UTI) for sepsis. Design. A systematic review and meta-analysis of randomized controlled trials (RCTs). Data Sources. The following databases: PubMed, Embase, and Cochrane Central were searched to identify related clinical trials. The search terms were “ulinastatin”, “thymosin”, and “sepsis”. Results. Six RCTs, 944 septic patients in total, were included in this meta-analysis. The result shows UTI increased the 28-day survival rate of septic patients, odds ratio (OR) = 2.01, 95% CI [1.53, 2.64]. After the treatment with UTI, the APACHE II score (four studies) dropped 4.72 further, mean = −4.72, 95% CI [−6.54, −2.91] (). The mean time of ICU stay (four studies) in UTI group decreased 3.03 days further, mean = −3.03 [−6.99, 0.95] (), and mechanical ventilation time (four studies) decreased 2.05 days, mean = −1.81 [−2.96, −0.66] (). With the treatment of UTI, CD4+T cells raised 5.13%, mean = 5.13, 95% CI [2.75, 7.50] (); there was no significant change in CD8+T cells, mean = −0.74 [−2.93, 1.45] (). Conclusion. According to this meta-analysis, with the treatment of UTI, the short-term survival rate of septic patients was increased and the illness severity was alleviated. ICU stay and mechanical ventilation time were effectively shortened. The beneficial effect of UTI might be due to the potential immunomodulatory effects of these two drugs. Feng Yun Wang, Bin Fang, Xin Hua Qiang, Tie Ou Yu, Jia Rong Zhong, Jun Cao, and Li Xin Zhou Copyright © 2016 Feng Yun Wang et al. All rights reserved. Paediatric Tuberculosis at a Referral Hospital in Istanbul: Analysis of 250 Cases Thu, 19 May 2016 09:49:25 +0000 Background. Tuberculosis (TB) still remains a growing public health problem globally. TB in children is often diagnosed clinically. Methods. We conducted a retrospective chart review of children with TB from November 2004 through December 2010 to determine the appropriateness of using contact history and diagnostic testing. Results. A total of 250 children with TB were identified. One hundred and sixty-two children had only pulmonary disease while 39 had features of both extrapulmonary and pulmonary TB. Mean age was 7.8 years. Thirty-six patients had known contacts. The index case/cases were first-degree relatives in 75%. Sixteen patients who were symptomless were yielded by contact investigation of newly identified TB cases. Tuberculin skin test positivity was 53.3%. Acid-fast bacilli smear positivity was 13.1%, and culture positivity was 18.7%. Twenty-six patients had histopathology of nonrespiratory specimens (lymph nodes and other tissues) showing granulomatous inflammation and caseous necrosis consistent with TB. Conclusions. Presence of contact history directed us to search for TB in children with nonspecific symptoms even if physical examinations were normal. Some children who were close contacts to TB cases were identified to have TB before development of symptoms. Ozden Turel, Selcen Kazanci, Ismail Gonen, Cigdem Aydogmus, Emel Karaoglan, and Rengin Siraneci Copyright © 2016 Ozden Turel et al. All rights reserved. Mycobacterium tuberculosis Contaminant Risk on Bone Marrow Aspiration Material from Iliac Bone Patients with Active Tuberculous Spondylitis Wed, 11 May 2016 07:50:06 +0000 There was a concern on Mycobacterium tuberculosis spreading to the bone marrow, when it was applied on tuberculous spine infection. This research aimed to study the probability of using autologous bone marrow as a source of mesenchymal stem cell for patients with tuberculous spondylitis. As many as nine patients with tuberculous spondylitis were used as samples. During the procedure, the vertebral lesion material and iliac bone marrow aspirates were obtained for acid fast staining, bacteria culture, and PCR (polymerase chain reaction) tests for Mycobacterium tuberculosis at the Clinical Microbiology Laboratory of Faculty of Medicine Universitas Indonesia. This research showed that there was a relationship between diagnostic confirmation of tuberculous spondylitis based on the PCR test and bacterial culture on the solid vertebral lesion material with the PCR test and bacterial culture from the bone marrow aspirates. If the diagnostic confirmation concluded positive results, then there was a higher probability that there would be a positive result for the bone marrow aspirates, so that it was not recommended to use autologous bone marrow as a source of mesenchymal stem cell for patients with tuberculous spondylitis unless the PCR and culture examination of the bone marrow showed a negative result. Ahmad Jabir Rahyussalim, Tri Kurniawati, and Andriansjah Rukmana Copyright © 2016 Ahmad Jabir Rahyussalim et al. All rights reserved. Prevalence and Influencing Factors of Thyroid Dysfunction in HIV-Infected Patients Wed, 20 Apr 2016 06:16:54 +0000 Thyroid dysfunction is more common in human immunodeficiency virus (HIV) patients. But the effects of highly active antiretroviral therapy (HAART) and hepatitis B/C virus (HBV/HCV) coinfection on thyroid function is unclear. We retrospectively reviewed the data of 178 HIV patients and determined the prevalence of thyroid dysfunction and the relationship between thyroid hormone levels, CD4 cell count, HIV-1 duration, HAART duration/regimens, and HBV/HCV coinfection. Of the 178 patients, 59 (33.1%) had thyroid dysfunction, mostly hypothyroidism. Thyroid dysfunction was significantly more frequent in the HAART group (41/104, 39.4%) than in the HAART-naïve group (18/74, 24.3%; ). The mean CD4 cell count was significantly lower in patients with hypothyroidism (372 ± 331/μL) than in the other patients (). The FT4 level was significantly lower in the HAART group than in the HAART-naïve group ( versus  pg/mL, ). FT3/FT4 levels were negatively related to HIV duration and FT3 levels were positively related to CD4 cell (). HBV patients had lower FT3 levels, while HCV patients had higher FT3 and FT4 levels (). Thyroid dysfunction is more common in HIV patients on HAART, mainly manifested as hypothyroidism. FT3/FT4 levels are correlated with HIV progression. HBV/HCV coinfection increases the probability of thyroid dysfunction. Shujing Ji, Changzhong Jin, Stefan Höxtermann, Wolfgang Fuchs, Tiansheng Xie, Xiangyun Lu, Haibo Wu, Linfang Cheng, Adriane Skaletz-Rorowski, Norbert H. Brockmeyer, and Nanping Wu Copyright © 2016 Shujing Ji et al. All rights reserved. Differences in Knowledge, Attitude, and Behavior towards HIV/AIDS and Sexually Transmitted Infections between Sexually Active Foreign and Chinese Medical Students Tue, 19 Apr 2016 14:05:29 +0000 Although the prevalence of human immunodeficiency virus (HIV) decreased in the last decade worldwide, the number of deaths due to HIV/AIDS and communicable diseases including syphilis, hepatitis, and tuberculosis had dramatically increased in developing countries. Education and behavior are incredibly important factors to prevent these diseases’ spread. This study highlights the range of differences in knowledge, attitude, and behavior of 434 sexually active medical students towards HIV/AIDS and sexually transmitted infections (STIs). Because the surveyed population constitutes the forefront of healthcare providers and was originated from different area of the world, this is the first time a study sought to investigate the behavioral attitude of this group of population irrespective of the three levels of their academic and professional knowledge. Several factors including sociodemographic characteristics, sexual behavior, HIV/AIDS, and STIs related patterns play a key role in medical student attitude and behavior towards people infected with HIV/AIDS and STIs. Our findings add consistent value in prior studies which aimed to stop new infections and also imply further investigations on the management of the studied infections by medical students. The present study arouses much interest among participants and provides evidence of reinforcing medical students’ education on HIV/AIDS and STIs. Martin Kuete, Qiao Huang, Abid Rashid, Xiu Lan Ma, HongFang Yuan, Juan Pablo Escalera Antezana, Rakhmanov Yeltay, Meng Rao, Qian He, ChengLiang Xiong, and HuiPing Zhang Copyright © 2016 Martin Kuete et al. All rights reserved. Does Citrulline Have Protective Effects on Liver Injury in Septic Rats? Tue, 19 Apr 2016 13:49:49 +0000 Citrulline (Cit) supplementation was proposed to serve as a therapeutic intervention to restore arginine (Arg) concentrations and improve related functions in sepsis. This study explored whether citrulline had positive effects on liver injury and cytokine release in the early stages of sepsis. The cecal ligation and puncture (CLP) model was utilized in our study. Rats were divided into four groups: normal, Cit, CLP, and CLP+Cit. The CLP group and CLP+Cit group were separated into 6-, 12-, and 24-hour groups, according to the time points of sacrifice after surgery. Intragastric administration of L-citrulline was applied to rats in Cit and CLP+Cit groups before surgery. Serum AST and ALT levels and levels of MDA, SOD, NO, and iNOS in the liver tissues were evaluated. Plasma concentrations of Cit and Arg were assessed using HPLC-MS/MS. Serum concentrations of cytokines and chemokines were calculated by Luminex. Results showed SOD activities of CLP+Cit groups were significantly higher than that of CLP groups, contrasting with the MDA and NO levels which were significantly lower in CLP+Cit groups than in CLP groups. In addition, plasma concentrations of TNF-α, IL-6, and IL-1β were significantly lower in the CLP+Cit 6-hour group than in the CLP 6-hour group. Bin Cai, Yu-long Luo, Shi-jun Wang, Wei-yuan Wei, Xue-hui Zhang, Wei Huang, Tong Li, Meng Zhang, Nan Wu, Gopaul Roodrajeetsing, and Sen Zhang Copyright © 2016 Bin Cai et al. All rights reserved. Infections due to Carbapenem Resistant Enterobacteriaceae among Saudi Arabian Hospitalized Patients: A Matched Case-Control Study Wed, 06 Apr 2016 06:58:19 +0000 Background. We conducted this case-control study to determine the risk factors and treatment outcome of infections due to carbapenem resistant Enterobacteriaceae in our institution. Methods. This is a matched case-control study of patients with infection due to carbapenem resistant Enterobacteriaceae (CRE) and carbapenem susceptible Enterobacteriaceae (CSE), from Riyadh, Saudi Arabia, between March 2012 and December 2013. Results. During this period, 29 cases and 58 controls were studied. The mean ages of the cases (55.4 years) and controls (54.7 years) were similar (). Cases had higher mean Charlson comorbidity index (CCI) (3.1) than controls (1.9), . Several factors contributed to infection among the studied population. Prior uses of piperacillin-tazobactam, a carbapenem, a quinolone, and metronidazole were significantly associated with CRE infections. Nine of the cases died compared with 7 of the controls, . Mortality was associated with advanced age, the presence of comorbidities, ICU stay, and receipt of invasive procedures. Conclusions. Infections due to CRE resulted in a significantly increased mortality. Combination antibiotic therapy was associated with reduced mortality. Properly designed randomized controlled studies are required to better characterize these findings. M. A. Garbati, H. Sakkijha, and A. Abushaheen Copyright © 2016 M. A. Garbati et al. All rights reserved. A Rapid In-Clinic Test Detects Acute Leptospirosis in Dogs with High Sensitivity and Specificity Thu, 24 Mar 2016 08:41:35 +0000 A rapid IgM-detection immunochromatographic test (WITNESS® Lepto, Zoetis) has recently become available to identify acute canine leptospirosis at the point of care. Diagnostic sensitivity and specificity of the test were evaluated by comparison with the microscopic agglutination assay (MAT), using a positive cut-off titer of ≥800. Banked serum samples from dogs exhibiting clinical signs and suspected leptospirosis were selected to form three groups based on MAT titer: (1) positive (); (2) borderline (); and (3) negative (). Using an analysis to weight group sizes to reflect French prevalence, the sensitivity and specificity were 98% and 93.5% (88.2% unweighted), respectively. This test rapidly identifies cases of acute canine leptospirosis with high levels of sensitivity and specificity with no interference from previous vaccination. Angeli Kodjo, Christophe Calleja, Michael Loenser, Dan Lin, and Joshua Lizer Copyright © 2016 Angeli Kodjo et al. All rights reserved. Serum Cytokine Responses over the Entire Clinical-Immunological Spectrum of Human Leishmania (L.) infantum chagasi Infection Tue, 08 Mar 2016 14:33:58 +0000 The clinical-immunological spectrum of human Leishmania (L.) infantum chagasi infection in Amazonian Brazil was recently reviewed based on clinical, DTH, and IFAT (IgG) evaluations that identified five profiles: three asymptomatic (asymptomatic infection, AI; subclinical resistant infection, SRI; and indeterminate initial infection, III) and two symptomatic (symptomatic infection, SI; American visceral leishmaniasis, AVL; and subclinical oligosymptomatic infection, SOI). TNF-α, IL-4, IL-6, and IL-10 serum cytokines were analyzed using multiplexed Cytometric Bead Array in 161 samples from endemic areas in the Brazilian Amazon: SI [AVL] (21 cases), III (49), SRI (19), SOI (12), AI (36), and a control group [CG] (24). The highest IL-6 serum levels were observed in the SI profile (AVL); higher IL-10 serum levels were observed in SI than in SOI or CG and in AI and III than in SOI; higher TNF-α serum levels were seen in SI than in CG. Positive correlations were found between IL-6 and IL-10 serum levels in the SI and III profiles and between IL-6 and TNF-α and between IL-4 and TNF-α in the III profile. These results provide strong evidence for associating IL-6 and IL-10 with the immunopathogenesis of AVL and help clarify the role of these cytokines in the infection spectrum. Patrícia Karla Ramos, Karina Inácio Carvalho, Daniela Santoro Rosa, Ana Paula Rodrigues, Luciana Vieira Lima, Marliane Batista Campos, Claudia Maria C. Gomes, Márcia Dalastra Laurenti, Carlos Eduardo Corbett, and Fernando Tobias Silveira Copyright © 2016 Patrícia Karla Ramos et al. All rights reserved. Determining the Independent Risk Factors and Mortality Rate of Nosocomial Infections in Pediatric Patients Mon, 15 Feb 2016 09:16:02 +0000 The objective of this study was to determine the rate, independent risk factors, and outcomes of healthcare-associated infections in pediatric patients. This study was performed between 2011 and 2014 in pediatric clinic and intensive care unit. 86 patients and 86 control subjects were included in the study. Of 86 patients with nosocomial infections (NIs), there were 100 NIs episodes and 90 culture growths. The median age was 32.0 months. The median duration of hospital stay of the patients was 30.0 days. The most frequent pathogens were Coagulase-negative Staphylococcus, Acinetobacter spp., Klebsiella spp., and Candida spp. Unconsciousness, prolonged hospitalization, transfusion, mechanical ventilation, use of central venous catheter, enteral feeding via a nasogastric tube, urinary catheter, and receiving carbapenems and glycopeptides were found to be significantly higher in NIs patients. Multivariate logistic regression analysis showed prolonged hospitalization, neutropenia, and use of central venous catheter and carbapenems as the independent risk factors for NIs. In the univariate analysis, unconsciousness, mechanical ventilation, enteral feeding, use of enteral feeding via a nasogastric tube, H2 receptor blockers, and port and urinary catheter were significantly associated with mortality. In the multiple logistic regression analysis, only mechanical ventilation was found as an independent predictor of mortality in patients with NIs. Fesih Aktar, Recep Tekin, Ali Güneş, Cevat Ülgen, İlhan Tan, Sabahattin Ertuğrul, Muhammet Köşker, Hasan Balık, Duran Karabel, and Ilyas Yolbaş Copyright © 2016 Fesih Aktar et al. All rights reserved. Epidemic Profile of Maternal Syphilis in China in 2013 Thu, 11 Feb 2016 12:05:04 +0000 Objective. The aim of this study was to investigate the epidemiological characteristics and adverse pregnancy outcomes of pregnant women with syphilis infection in China. Methods. Data were from China’s Information System of Prevention of Mother-to-Child Transmission of Syphilis Management. Women who were registered in the system and delivered in 2013 were included in the analysis. Results. A total of 15884 pregnant women with syphilis infection delivered in China in 2013. 79.1% of infected women attended antenatal care at or before 37 gestational weeks; however, 55.4% received no treatment or initiated the treatment after 37 gestational weeks. 14.0% of women suffered serious adverse pregnancy outcomes including stillbirth/neonatal death, preterm delivery/low birth weight, or congenital syphilis in newborns. High maternal titer (≥1 : 64) and late treatment (>37 gestational weeks)/nontreatment were significantly associated with increased risk of congenital syphilis and the adjusted ORs were 1.88 (95% CI 1.27 to 2.80) and 3.70 (95% CI 2.36 to 5.80), respectively. Conclusion. Syphilis affects a great number of pregnant women in China. Large proportions of women are not detected and treated at an early pregnancy stage. Burden of adverse pregnancy outcomes is high among infected women. Comprehensive interventions still need to be strengthened to improve uptake of screening and treatment for maternal syphilis. Lixia Dou, Xiaoyan Wang, Fang Wang, Qian Wang, Yaping Qiao, Min Su, Xi Jin, Jie Qiu, Li Song, and Ailing Wang Copyright © 2016 Lixia Dou et al. All rights reserved. Prevalence and Clinical Characteristics of Mycobacterial Diseases in the Barletta-Andria-Trani Province, Italy (2005–2013) Sun, 17 Jan 2016 10:56:03 +0000 Tuberculosis remains one of the major worldwide problems regarding public health. This study evaluates the burden of this disease in the BAT Province of the Apulia region (Italy); 12,295 patients were studied, including 310 immigrants. Tubercular disease and mycobacteriosis were found in 129 patients. The number of new TB cases/year ranged from three in 2005 to 12 in 2009. TB was more frequently localized in the lung (70.5%). 14.4% of cases were institutionalized patients for severe neurological and/or psychiatric disease. The database evidenced certain aspects of our study population: the large number of TB patients institutionalized between natives, but no larger presence of TB among HIV-positive patients in immigrants compared to Italians. Our findings should help to redefine the alarm regarding the spread of an epidemical form of TB but also to present certain criticisms regarding patient management (especially immigrants) regarding costs, hospitalization, and difficulty of reinstating the patient in the community. Further our data underscore the importance of prevalence of TB in bedridden, institutionalized patients. Gaetano Brindicci, Carmen Rita Santoro, Giovanna Trillo, Anna Volpe, Daniela Loconsole, Laura Monno, and Tommaso Fontana Copyright © 2016 Gaetano Brindicci et al. All rights reserved. Prevalence and Predictors of Self-Medication with Antibiotics in Al Wazarat Health Center, Riyadh City, KSA Tue, 05 Jan 2016 13:03:27 +0000 Background. Antibiotics are responsible for most dramatic improvement in medical therapy in history. These medications contributed significantly to the decreasing mortality and morbidity when prescribed based on evidence of microbial infection. Objective. The aim of this study was to determine the prevalence and predictors of self-prescription with antibiotics in Al Wazarat Health Center, Riyadh City, Kingdom of Saudi Arabia. Material and Methods. Cross-sectional study was conducted in Al Wazarat Health Center between February 2014 and November 2014. Respondents were randomly selected using a multistage clustered random sampling technique. Data was entered into SPSS version 21 and analyzed. Descriptive statistics and multiple logistic regression models were applied. Results. A total of 681 patients have participated in this study with a response rate of 92%. The prevalence of self-prescription with antibiotics in Al Wazarat Health Center was 78.7%. Amoxicillin was the most used self-prescribed antibiotic with prevalence of (22.3%). Friend advice on self-prescription of antibiotics use () and pharmacy near to the participants () were the most common predictors for self-prescription with antibiotics. Conclusion. The level of self-prescribing antibiotics is relatively high among participants. Health education on the appropriate use of antibiotics is highly recommended. The proper use of treatment guidelines for antibiotic therapy will significantly reduce self-prescription with antibiotics. Abdulrahman Al Rasheed, Umar Yagoub, Hesham Alkhashan, Osama Abdelhay, Ahmad Alawwad, Aboud Al Aboud, and Saad Al Battal Copyright © 2016 Abdulrahman Al Rasheed et al. All rights reserved. Comment on “Is There a Risk of Yellow Fever Virus Transmission in South Asian Countries with Hyperendemic Dengue?” Sun, 29 Nov 2015 11:04:15 +0000 John T. Cathey Copyright © 2015 John T. Cathey. All rights reserved. Clinical and Etiological Characteristics of Atypical Hand-Foot-and-Mouth Disease in Children from Chongqing, China: A Retrospective Study Thu, 26 Nov 2015 12:56:30 +0000 Background. Hand-foot-and-mouth disease (HFMD) is a disease that had similar manifestations to chickenpox, impetigo, and measles, which is easy to misdiagnose and subsequently causes delayed therapy and subsequent epidemic. To date, no study has been conducted to report the clinical and epidemiological characteristics of atypical HFMD. Methods. 64 children with atypical HFMD out of 887 HFMD children were recruited, stool was collected, and viral VP1 was detected. Results. The atypical HFMD accounted for 7.2% of total HFMD in the same period (64/887) and there were two peaks in its prevalence in nonepidemic seasons. Ten children (15.6%) had manifestations of neurologic involvement, of whom 4 (6.3%) were diagnosed with severe HFMD and 1 with critically severe HFMD, but all recovered smoothly. Onychomadesis and desquamation were found in 14 patients (21.9%) and 15 patients (23.4%), respectively. The most common pathogen was coxsackievirus A6 (CV-A6) which accounted for 67.2%, followed by nontypable enterovirus (26.6%), enterovirus 71 (EV-A71) (4.7%), and coxsackievirus A16 (A16) (1.5%). Conclusions. Atypical HFMD has seasonal prevalence. The manifestations of neurologic involvement in atypical HFMD are mild and usually have a good prognosis. CV-A6 is a major pathogen causing atypical HFMD, but not a major pathogen in Chongqing, China. Xiang Yan, Zhen-Zhen Zhang, Zhen-Hua Yang, Chao-Min Zhu, Yun-Ge Hu, and Quan-Bo Liu Copyright © 2015 Xiang Yan et al. All rights reserved. Design and Synthesis of a Novel Cationic Peptide with Potent and Broad-Spectrum Antimicrobial Activity Wed, 25 Nov 2015 12:29:16 +0000 Antibacterial and antifungal peptides have increasingly been used to combat the antibiotic-resistant microbes in recent years. KW-13, a novel cationic α-helical antibacterial peptide consisting of 13 amino acid residues, was designed and chemically synthesized. The peptide has a net charge of +6 with a total hydrophobic ratio of 38%. The antibacterial experiments revealed that KW-13 strongly inhibited the growth of human pathogenic bacteria with minimal inhibitory concentrations of 4 and 16 μg/mL for Staphylococcus epidermidis and Staphylococcus aureus, respectively, while the hemolytic assay showed that this peptide did not destroy human red blood cells in vitro. Scanning electron microscopy imaging of Escherichia coli confirmed that KW-13 can damage the membrane of bacterial cells. Thus, this peptide could be a potential candidate for the treatment of infectious diseases. Wen-Ping Liu, Ya-Hui Chen, Xin Ming, and Yi Kong Copyright © 2015 Wen-Ping Liu et al. All rights reserved. MicroRNA-381 Negatively Regulates TLR4 Signaling in A549 Cells in Response to LPS Stimulation Tue, 24 Nov 2015 12:12:21 +0000 It is widely reported that miR-381 is dysregulated in various tumors. However, the specific role of miR-381 in respiratory infections has not been reported. To probe this role, A549 cells were pretreated with 1 μg/mL LPS for 24 h. The level of miR-381 was detected using RT-qPCR. The expression of proinflammatory cytokines was determined using an ELISA kit and western blotting. Bioinformatics analysis was used to predict the target genes of miR-381, and a luciferase reporter assay was used to validate the expression of the target genes. miR-381 expression was increased in A549 cells treated with LPS, which is a ligand of TLRs. Further study revealed that the overexpression of miR-381 increased the activity of NF-κB signaling, thereby increasing the expression of IL-6, TNFα, and COX-2. Further study revealed that IκBα was a target gene of miR-381. The upregulation of miR-381 under LPS stimulation contributes to respiratory infections mainly by targeting IκBα. Zhihao Xu, Dapeng Dong, Xiaofei Chen, Huaqiong Huang, and Shenglan Wen Copyright © 2015 Zhihao Xu et al. All rights reserved. A Novel Insight into Dehydroleucodine Mediated Attenuation of Pseudomonas aeruginosa Virulence Mechanism Wed, 11 Nov 2015 13:41:29 +0000 Increasing resistance of Pseudomonas aeruginosa (P. aeruginosa) to conventional treatments demands the search for novel therapeutic strategies. In this study, the antimicrobial activity of dehydroleucodine (DhL), a sesquiterpene lactone obtained from Artemisia (A.) douglasiana, was screened against several pathogenic virulence effectors of P. aeruginosa. In vitro, minimum inhibitory concentration of DhL was determined against P. aeruginosa strains PAO1, PA103, PA14, and multidrug resistant clinical strain, CDN118. Results showed that DhL was active against each strain where PAO1 and PA103 showed higher susceptibility (MIC 0.48 mg/mL) as compared to PA14 (MIC 0.96 mg/mL) and CDN118 (MIC 0.98 mg/mL). Also, when PAO1 strain was grown in the presence of DhL (MIC50, 0.12 mg/mL), a delay in the generation time was noticed along with significant inhibition of secretory protease and elastase activities, interruption in biofilm attachment phase in a stationary culture, and a significant decline in Type III effector ExoS. At MIC50, DhL treatment increased the sensitivity of P. aeruginosa towards potent antibiotics. Furthermore, treatment of P. aeruginosa with DhL prevented toxin-induced apoptosis in macrophages. These observations suggest that DhL activity was at the bacterial transcriptional level. Hence, antimicrobial activity of DhL may serve as leads in the development of new anti-Pseudomonas pharmaceuticals. S. Mustafi, M. L. Veisaga, L. A. López, and M. A. Barbieri Copyright © 2015 S. Mustafi et al. All rights reserved. Macrocolonies (Granules) Formation as a Cause of False-Negative Results in the MGIT 960 System: Cause Analysis and Correlation with Mycobacterial Species Wed, 04 Nov 2015 09:17:38 +0000 Background. The viable mycobacterial bacilli can sometimes form granules in the Mycobacterium Growth Indicator Tube (MGIT) to produce instrument-negative outcomes when BACTEC MGIT 960 culture is performed. The cause of this phenomenon has never been analyzed. Methods. Thirty-one instrument-negative, granule presenting MGIT vials were collected for conducting acid-fast staining and also liquid and solid subculture. Species identification and drug susceptibility test were performed with the recovered strains. Cultivation test was done by inoculating small amount of bacilli into the MGIT vials. Results. Twenty-four and twenty-nine of the tested MGIT vials were smear and culture positive, respectively. In total, 18, 4, and 7 of the cultivated strains were identified as Mycobacterium tuberculosis complex, M. intracellulare, and M. xenopi, respectively. When a limited amount of bacilli was inoculated, the granule formation was observed for M. xenopi strains in the MGIT system. Conclusions. The granules observed in the instrument-negative MGIT vials consisted of viable bacilli, which emphasized the need of visual inspection to increase recovery rate. Limited bacterial load and specific species might be the cause of granule forming. Xia Yu, Liping Zhao, Guanglu Jiang, Yifeng Ma, Weimin Li, and Hairong Huang Copyright © 2015 Xia Yu et al. All rights reserved. Thyroid Peroxidase Antibodies in Non-Interferon Treated Hepatitis C Patients in Pakistan Tue, 03 Nov 2015 09:18:23 +0000 Objective. Interferon therapy of HCV infected patients is associated with development of thyroid dysfunctions. Patients with pretreatment presence of antithyroid peroxidase (TPO-Ab) are at greater risk. This study, probably the first in Pakistan, was planned to determine TPO-Ab in sera of treatment-naive local HCV patients. Setting. Centre for Nuclear Medicine (CENUM), Mayo Hospital, Lahore. Patients and Methods. During July to December 2012, 190 patients (140 females, 50 males) newly diagnosed for HCV infection were selected for this study. Their age range was 15–55 years (mean: 35.3 ± 9.1 years). 262 age matched healthy subjects (211 females and 50 males) were recruited as control. Serum-free thyroxin (FT4) and thyroid stimulating hormone (TSH) were detected by radioimmunoassay techniques. Serum TPO-Ab titer was determined by ELISA method using commercial kits. Results. Serum FT4 and TSH levels in HCV patients and controls were within normal range. Between two groups there was no significant difference in mean value of FT4 (16.0 ± 3.0 versus 16.2 ± 3.9; ) but mean TSH value was significantly lower in HCV patients (1.5 ± 0.8 versus 1.8 ± 0.9; ). Among HCV patients 51 (26.8%) were TPO-Ab positive and among control subjects 18 (6.9%) were TPO-Ab positive. The difference was statistically significant . Further analysis showed that among HCV patients 39 (27.8%) females and 12 (24.0%) males were TPO-Ab positive, respectively, and difference was not statistically significant . Moreover, TPO-Ab positive patients were older and had significantly higher serum TSH as compared to TPO-Ab negative HCV patients. Conclusion. Independent of patient’s gender and increasing with advancing age, about one-fourth of local untreated HCV patients are TPO-Ab positive and are at greater risk of developing thyroid disorders during and after interferon treatment. Muhammad Imtiaz Shafiq, Amna Gauhar, Muhammad Akram, and Shan Elahi Copyright © 2015 Muhammad Imtiaz Shafiq et al. All rights reserved. The Evolving Genotypic Profile of HIV-1 Mutations Related to Antiretroviral Treatment in the North Region of Brazil Mon, 12 Oct 2015 14:16:53 +0000 HIV related mutations can be associated with decreased susceptibility to antiretrovirals and treatment failures. There is scarce information about HIV mutations in persons failing HIV treatment in North of Brazil. Our aim was to evaluate evolution of HIV subtypes and mutations patterns related to antiretroviral therapy in this region. We investigated HIV resistance profile in adults failing antiretroviral regimen in Northern Brazil from January, 2004, through December, 2013. Genotype data was evaluated through Stanford University algorithm. There were 377 genotypes from different individuals to evaluate. Resistance mutations were similar to worldwide reports and related to antiretroviral exposure. Most prevalent mutations in the reverse transcriptase gene were M184V (80.1%) and K130N (40.6%). Thymidine associated mutations were more frequent in multiexperienced patients. Most common protease mutations were M46I, V82A, I54V, L90M, I84V, M46L, and L76V. Subtype B was the most prevalent (90.7%). There were differences between subtypes B and non-B mutations. We documented for the first time subtypes and patterns of HIV associated mutations in Northern Brazil. A1 subtype was identified for the first time in this area. Depending on drug regimen and how experienced the patient is, an empirical switch of a failing antiretroviral treatment could be a reasonable option. Carmen Andréa F. Lopes, Marcelo A. Soares, Diego R. Falci, and Eduardo Sprinz Copyright © 2015 Carmen Andréa F. Lopes et al. All rights reserved. Successive Intramuscular Boosting with IFN-Alpha Protects Mycobacterium bovis BCG-Vaccinated Mice against M. lepraemurium Infection Mon, 21 Sep 2015 14:10:52 +0000 Leprosy caused by Mycobacterium leprae primarily affects the skin and peripheral nerves. As a human infectious disease, it is still a significant health and economic burden on developing countries. Although multidrug therapy is reducing the number of active cases to approximately 0.5 million, the number of cases per year is not declining. Therefore, alternative host-directed strategies should be addressed to improve treatment efficacy and outcome. In this work, using murine leprosy as a model, a very similar granulomatous skin lesion to human leprosy, we have found that successive IFN-alpha boosting protects BCG-vaccinated mice against M. lepraemurium infection. No difference in the seric isotype and all IgG subclasses measured, neither in the TH1 nor in the TH2 type cytokine production, was seen. However, an enhanced iNOS/NO production in BCG-vaccinated/i.m. IFN-alpha boosted mice was observed. The data provided in this study suggest a promising use for IFN-alpha boosting as a new prophylactic alternative to be explored in human leprosy by targeting host innate cell response. G. G. Guerrero, J. Rangel-Moreno, S. Islas-Trujillo, and Ó. Rojas-Espinosa Copyright © 2015 G. G. Guerrero et al. All rights reserved. Comment on “Nontuberculous Mycobacteria Isolation from Clinical and Environmental Samples in Iran: Twenty Years of Surveillance” Tue, 08 Sep 2015 11:58:50 +0000 Eduardo Hernández-Garduño Copyright © 2015 Eduardo Hernández-Garduño. All rights reserved. Mathematical Model of Innate and Adaptive Immunity of Sepsis: A Modeling and Simulation Study of Infectious Disease Tue, 08 Sep 2015 09:08:09 +0000 Sepsis is a systemic inflammatory response (SIR) to infection. In this work, a system dynamics mathematical model (SDMM) is examined to describe the basic components of SIR and sepsis progression. Both innate and adaptive immunities are included, and simulated results in silico have shown that adaptive immunity has significant impacts on the outcomes of sepsis progression. Further investigation has found that the intervention timing, intensity of anti-inflammatory cytokines, and initial pathogen load are highly predictive of outcomes of a sepsis episode. Sensitivity and stability analysis were carried out using bifurcation analysis to explore system stability with various initial and boundary conditions. The stability analysis suggested that the system could diverge at an unstable equilibrium after perturbations if (maximum release rate of Tumor Necrosis Factor- (TNF-) α by neutrophil) falls below a certain level. This finding conforms to clinical findings and existing literature regarding the lack of efficacy of anti-TNF antibody therapy. Zhenzhen Shi, Chih-Hang J. Wu, David Ben-Arieh, and Steven Q. Simpson Copyright © 2015 Zhenzhen Shi et al. All rights reserved. Computational and Theoretical Analysis of Human Diseases Associated with Infectious Pathogens Tue, 08 Sep 2015 06:26:30 +0000 Suares Clovis Oukouomi Noutchie, Cedrick Aurelien Kitio Kwuimy, Jean Jules Tewa, Farai Nyabadza, and Necdet Bildik Copyright © 2015 Suares Clovis Oukouomi Noutchie et al. All rights reserved. Changes in B-Cell Counts and Percentages during Primary HIV Infection Associated with Disease Progression in HIV-Infected Men Who Have Sex with Men: A Preliminary Study Mon, 07 Sep 2015 06:12:09 +0000 Numerous anomalies in B-cell phenotypes and functions have been described in HIV-infected individuals. However, the actual relationship between B cells and disease progression remains unclear. In this study, we investigated B-cell counts/percentages during a 12-month infection period in HIV-infected individuals that eventually developed into typical progressors (TPs) or rapid progressors (RPs). We found, after 12 months of infection, the baseline B-cell counts/percentages correlated positively with CD4+ T-cell counts ( and ) and negatively with HIV viral set points ( and ). Kaplan-Meier survival analysis showed that high baseline B-cell counts/percentages were associated with a slow CD4-cell decline. B-cell kinetics indicated the baseline B-cell counts/percentages could be factors distinguishing between TPs and RPs. The combination of the baseline B-cell counts and percentages was associated with rapid disease progression (a 80.7% predictive value as measured by the area under the curve). These results indicate that the baseline B-cell counts/percentages might be associated with HIV disease progression. Chen Cui, Yongjun Jiang, Zining Zhang, Qinghai Hu, Zhenxing Chu, Junjie Xu, Bin Zhao, Haibo Ding, Jing Liu, Xiaoxu Han, Yaming Cao, and Hong Shang Copyright © 2015 Chen Cui et al. All rights reserved. A Bayesian Outbreak Detection Method for Influenza-Like Illness Sun, 06 Sep 2015 13:22:02 +0000 Epidemic outbreak detection is an important problem in public health and the development of reliable methods for outbreak detection remains an active research area. In this paper we introduce a Bayesian method to detect outbreaks of influenza-like illness from surveillance data. The rationale is that, during the early phase of the outbreak, surveillance data changes from autoregressive dynamics to a regime of exponential growth. Our method uses Bayesian model selection and Bayesian regression to identify the breakpoint. No free parameters need to be tuned. However, historical information regarding influenza-like illnesses needs to be incorporated into the model. In order to show and discuss the performance of our method we analyze synthetic, seasonal, and pandemic outbreak data. Yury E. García, J. Andrés Christen, and Marcos A. Capistrán Copyright © 2015 Yury E. García et al. All rights reserved. The Implications of HIV Treatment on the HIV-Malaria Coinfection Dynamics: A Modeling Perspective Sun, 06 Sep 2015 13:20:27 +0000 Most hosts harbor multiple pathogens at the same time in disease epidemiology. Multiple pathogens have the potential for interaction resulting in negative impacts on host fitness or alterations in pathogen transmission dynamics. In this paper we develop a mathematical model describing the dynamics of HIV-malaria coinfection. Additionally, we extended our model to examine the role treatment (of malaria and HIV) plays in altering populations’ dynamics. Our model consists of 13 interlinked equations which allow us to explore multiple aspects of HIV-malaria transmission and treatment. We perform qualitative analysis of the model that includes positivity and boundedness of solutions. Furthermore, we evaluate the reproductive numbers corresponding to the submodels and investigate the long term behavior of the submodels. We also consider the qualitative dynamics of the full model. Sensitivity analysis is done to determine the impact of some chosen parameters on the dynamics of malaria. Finally, numerical simulations illustrate the potential impact of the treatment scenarios and confirm our analytical results. F. Nyabadza, B. T. Bekele, M. A. Rúa, D. M. Malonza, N. Chiduku, and M. Kgosimore Copyright © 2015 F. Nyabadza et al. All rights reserved. Fascioliasis: An Ongoing Zoonotic Trematode Infection Mon, 31 Aug 2015 11:59:26 +0000 Zoonotic trematode infections are an area of the neglected tropical diseases that have become of major interest to global and public health due to their associated morbidity. Human fascioliasis is a trematode zoonosis of interest in public health. It affects approximately 50 million people worldwide and over 180 million are at risk of infection in both developed and underdeveloped countries. The one health paradigm is an area that seeks to address the problem of zoonotic infections through a comprehensive and sustainable approach. This review attempts to address the major challenges in managing human and animal fascioliasis with valuable insights gained from the one health paradigm to global health and multidisciplinary integration. Mramba Nyindo and Abdul-Hamid Lukambagire Copyright © 2015 Mramba Nyindo and Abdul-Hamid Lukambagire. All rights reserved. The Absence of N-Acetyl-D-glucosamine Causes Attenuation of Virulence of Candida albicans upon Interaction with Vaginal Epithelial Cells In Vitro Thu, 20 Aug 2015 11:52:00 +0000 To better understand the molecular events underlying vulvovaginal candidiasis, we established an in vitro system. Immortalized vaginal epithelial cells were infected with live, yeast form C. albicans and C. albicans cultured in the same medium without vaginal epithelial cells were used as control. In both cases a yeast to hyphae transition was robustly induced. Whole transcriptome sequencing was used to identify specific gene expression changes in C. albicans. Numerous genes leading to a yeast to hyphae transition and hyphae specific genes were upregulated in the control hyphae and the hyphae in response to vaginal epithelial cells. Strikingly, the GlcNAc pathway was exclusively triggered by vaginal epithelial cells. Functional analysis in our in vitro system revealed that the GlcNAc biosynthesis is involved in the adherence to, and the ability to kill, vaginal epithelial cells in vitro, thus indicating the key role for this pathway in the virulence of C. albicans upon vulvovaginal candidiasis. Máté Manczinger, Alexandra Bocsik, Gabriella F. Kocsis, Andrea Vörös, Zoltán Hegedűs, Lilla Ördögh, Éva Kondorosi, Annamária Marton, Csaba Vízler, Vilmos Tubak, Mária Deli, Lajos Kemény, István Nagy, and Lóránt Lakatos Copyright © 2015 Máté Manczinger et al. All rights reserved. Interaction between Flavivirus and Cytoskeleton during Virus Replication Mon, 10 Aug 2015 12:34:40 +0000 Flaviviruses are potentially human pathogens that cause major epidemics worldwide. Flavivirus interacts with host cell factors to form a favourable virus replication site. Cell cytoskeletons have been observed to have close contact with flaviviruses, which expands the understanding of cytoskeleton functions during virus replication, although many detailed mechanisms are still unclear. The interactions between the virus and host cytoskeletons such as actin filaments, microtubules, and intermediate filaments have provided insight into molecular alterations during the virus infection, such as viral entry, in-cell transport, scaffold assembly, and egress. This review article focuses on the utilization of cytoskeleton by Flavivirus and the respective functions during virus replication. Kar Yue Foo and Hui-Yee Chee Copyright © 2015 Kar Yue Foo and Hui-Yee Chee. All rights reserved. In Silico Identification of Highly Conserved Epitopes of Influenza A H1N1, H2N2, H3N2, and H5N1 with Diagnostic and Vaccination Potential Thu, 06 Aug 2015 12:36:05 +0000 The unpredictable, evolutionary nature of the influenza A virus (IAV) is the primary problem when generating a vaccine and when designing diagnostic strategies; thus, it is necessary to determine the constant regions in viral proteins. In this study, we completed an in silico analysis of the reported epitopes of the 4 IAV proteins that are antigenically most significant (HA, NA, NP, and M2) in the 3 strains with the greatest world circulation in the last century (H1N1, H2N2, and H3N2) and in one of the main aviary subtypes responsible for zoonosis (H5N1). For this purpose, the HMMER program was used to align 3,016 epitopes reported in the Immune Epitope Database and Analysis Resource (IEDB) and distributed in 34,294 stored sequences in the Pfam database. Eighteen epitopes were identified: 8 in HA, 5 in NA, 3 in NP, and 2 in M2. These epitopes have remained constant since they were first identified (~91 years) and are present in strains that have circulated on 5 continents. These sites could be targets for vaccination design strategies based on epitopes and/or as markers in the implementation of diagnostic techniques. José Esteban Muñoz-Medina, Carlos Javier Sánchez-Vallejo, Alfonso Méndez-Tenorio, Irma Eloísa Monroy-Muñoz, Javier Angeles-Martínez, Andrea Santos Coy-Arechavaleta, Clara Esperanza Santacruz-Tinoco, Joaquín González-Ibarra, Yu-Mei Anguiano-Hernández, César Raúl González-Bonilla, Eva Ramón-Gallegos, and José Alberto Díaz-Quiñonez Copyright © 2015 José Esteban Muñoz-Medina et al. All rights reserved. Blood or Urine IP-10 Cannot Discriminate between Active Tuberculosis and Respiratory Diseases Different from Tuberculosis in Children Wed, 05 Aug 2015 10:28:29 +0000 Objectives. Interferon-γ inducible protein 10 (IP-10), either in blood or in urine, has been proposed as a tuberculosis (TB) biomarker for adults. This study aims to evaluate the potential of IP-10 diagnostics in children from Uganda, a high TB-endemic country. Methods. IP-10 was measured in the blood and urine concomitantly taken from children who were prospectively enrolled with suspected active TB, with or without HIV infection. Clinical/microbiological parameters and commercially available TB-immune assays (tuberculin skin test (TST) and QuantiFERON TB-Gold In-Tube (QFT-IT)) were concomitantly evaluated. Results. One hundred twenty-eight children were prospectively enrolled. The analysis was performed on 111 children: 80 (72%) of them were HIV-uninfected and 31 (27.9%) were HIV-infected. Thirty-three healthy adult donors (HAD) were included as controls. The data showed that IP-10 is detectable in the urine and blood of children with active TB, independent of HIV status and age. However, although IP-10 levels were higher in active TB children compared to HAD, the accuracy of identifying “active TB” was low and similar to the TST and QFT-IT. Conclusion. IP-10 levels are higher in children with respiratory illness compared to controls, independent of “TB status” suggesting that the evaluation of this parameter can be used as an inflammatory marker more than a TB test. Linda Petrone, Angela Cannas, Francesco Aloi, Martin Nsubuga, Joseph Sserumkuma, Ritah Angella Nazziwa, Levan Jugheli, Tedson Lukindo, Enrico Girardi, Klaus Reither, and Delia Goletti Copyright © 2015 Linda Petrone et al. All rights reserved. Comparison of the Effect of Vaginal Zataria multiflora Cream and Oral Metronidazole Pill on Results of Treatments for Vaginal Infections including Trichomoniasis and Bacterial Vaginosis in Women of Reproductive Age Tue, 21 Jul 2015 12:39:55 +0000 Effect of Zataria multiflora on bacterial vaginosis and Trichomonas vaginalis is shown in vivo and in vitro. We compare the effectiveness of Zataria multiflora cream and oral metronidazole pill on results of treatment for vaginal infections including Trichomonas and bacterial vaginosis; these infections occur simultaneously. The study included 420 women with bacterial vaginosis, Trichomonas vaginalis, or both infections together, who were randomly divided into six groups. Criteria for diagnosis were wet smear and Gram stain. Vaginal Zataria multiflora cream and placebo pill were administered to the experiment groups; the control group received oral metronidazole pill and vaginal placebo cream. Comparison of the clinical symptoms showed no significant difference in all three vaginitis groups receiving metronidazole pill and vaginal Zataria multiflora cream. However, comparison of the wet smear test results was significant in patients with trichomoniasis and bacterial vaginosis associated with trichomoniasis in the two treatment groups ( and ). Vaginal Zataria multiflora cream had the same effect of oral metronidazole tablets in improving clinical symptoms of all three vaginitis groups, as well as the treatment for bacterial vaginosis. It can be used as a drug for treatment of bacterial vaginosis and elimination of clinical symptoms of Trichomonas vaginitis. Khadijeh Abdali, Leila Jahed, Sedigheh Amooee, Mahnaz Zarshenas, Hamidreza Tabatabaee, and Reza Bekhradi Copyright © 2015 Khadijeh Abdali et al. All rights reserved. Utility of C-Reactive Protein Levels for Early Prediction of Dengue Severity in Adults Sun, 12 Jul 2015 11:27:20 +0000 Dengue has broad clinical presentation with unpredictable clinical evolution and outcome. We aimed to evaluate the utility of C-reactive protein (CRP) levels for distinguishing between mild and severe cases in the early phase of the dengue illness. We retrospectively evaluated adults with dengue from 2006 to 2014, according to 1997 and 2009 World Health Organization (WHO) criteria for severity. Of 191 included patients, 32.9% had nonshock dengue hemorrhagic fever (DHF), 3.1% dengue shock syndrome (DSS), and 7.9% severe dengue. The risk of DHF/DSS and severe dengue is significantly related to the increasing levels of CRP. Of 191 patients, 97 had CRP levels measured during the febrile (days 1–3); 85 during the critical (days 4–6); and 9 during the convalescent (days 7–10) illness phases. During the febrile phase, there was significant higher CRP level for DSS versus DF/nonshock DHF and severe dengue versus nonsevere dengue, with CRP cutoff level 30.1 mg/L (area under the receiver operating characteristic curve (AUC), 0.938; 100% sensitivity, 76.3% specificity) and 24.2 mg/L (AUC, 0.717; 70% sensitivity, 71.3% specificity), respectively. Our study highlights the utility of the CRP levels in early prediction of DSS and severe dengue in adult patients. Chien-Chih Chen, Ing-Kit Lee, Jien-Wei Liu, Shi-Yu Huang, and Lin Wang Copyright © 2015 Chien-Chih Chen et al. All rights reserved. The Effect of Latent Toxoplasma gondii Infection on the Immune Response in HIV-Infected Patients Sun, 12 Jul 2015 09:27:47 +0000 A relationship between latent toxoplasmosis and the immune system during HIV disease is poorly understood. Therefore, the aim of this follow-up study was to characterize immunological parameters in HIV-infected patients with latent toxoplasmosis and noninfected individuals. A total of 101 HIV-infected patients were enrolled in the study. The patients were classified into two groups based on anti-Toxoplasma gondii antibodies: a group of 55 toxoplasma-positive persons (TP) and a group of 46 toxoplasma-negative persons (TN). Absolute counts of several lymphocyte subsets decreased in the TP group, namely, T cells (), B cells (), NK cells (), CD4 T cells (), and CD8 T cells (). On the other hand, the percentage of CD8 T cells expressing CD38 and HLA-DR significantly increased during the follow-up in the TP group (, , resp.) as well as the intensity of CD38 and HLA-DR expression (MFI) on CD8 T cells (, , resp.). In the TN group, analysis of the kinetics of immunological parameters revealed no significant changes over time. In conclusion, the results suggest that latent T. gondii infection modulates the immune response during HIV infection. Ondrej Beran, Petr Kodym, Marek Maly, Alzbeta Davidova, Gabriela Reinvartova, David Jilich, Michal Holub, and Hanus Rozsypal Copyright © 2015 Ondrej Beran et al. All rights reserved. Seroepidemiology of Leptospira Exposure in General Population in Rural Durango, Mexico Thu, 09 Jul 2015 14:25:34 +0000 The magnitude of Leptospira exposure in rural Mexico is largely unknown. We sought to determine the seroprevalence of Leptospira IgG antibodies in adults in rural Durango, Mexico, and to determine the sociodemographic, behavioral, and housing characteristics of the subjects associated with Leptospira seropositivity. We performed a cross-sectional study in 282 adults living in rural Durango, Mexico. Sera from participants were analyzed for Leptospira IgG antibodies using a commercially available enzyme immunoassay. Seroprevalence association with the characteristics of the subjects was analyzed by bivariate and multivariate analyses. Of the 282 rural subjects (42.91 ± 17.53 years old) studied, 44 (15.6%) had anti-Leptospira IgG antibodies. Seropositivity to Leptospira was not associated with gender, educational level, employment, socioeconomic status, contact with animals or soil, or type of floors at home. In contrast, multivariate analysis showed that Leptospira exposure was associated with national trips (OR = 2.09; 95% CI: 1.05–4.16; ) and poor education of the head of the family (OR = 2.96; 95% CI: 1.51–5.78; ). We demonstrated serological evidence of Leptospira exposure in adults in rural northern Mexico. The contributing factors associated with Leptospira exposure found in the present study may be useful for optimal planning of preventive measures against Leptospira infection. Cosme Alvarado-Esquivel, Luis Francisco Sánchez-Anguiano, and Jesús Hernández-Tinoco Copyright © 2015 Cosme Alvarado-Esquivel et al. All rights reserved. Diversity and Global Distribution of IncL/M Plasmids Enabling Horizontal Dissemination of β-Lactam Resistance Genes among the Enterobacteriaceae Wed, 08 Jul 2015 11:14:54 +0000 Antibiotic resistance determinants are frequently associated with plasmids and other mobile genetic elements, which simplifies their horizontal transmission. Several groups of plasmids (including replicons of the IncL/M incompatibility group) were found to play an important role in the dissemination of resistance genes encoding β-lactamases. The IncL/M plasmids are large, broad host range, and self-transmissible replicons. We have identified and characterized two novel members of this group: pARM26 (isolated from bacteria inhabiting activated sludge from a wastewater treatment plant) and pIGT15 (originating from a clinical strain of Escherichia coli). This instigated a detailed comparative analysis of all available sequences of IncL/M plasmids encoding β-lactamases. The core genome of these plasmids is comprised of 20 genes with conserved synteny. Phylogenetic analyses of these core genes allowed clustering of the plasmids into four separate groups, which reflect their antibiotic resistance profiles. Examination of the biogeography of the IncL/M plasmids revealed that they are most frequently found in bacteria of the family Enterobacteriaceae originating from the Mediterranean region and Western Europe and that they are able to persist in various ecological niches even in the absence of direct antibiotic selection pressure. Marcin Adamczuk, Piotr Zaleski, Lukasz Dziewit, Renata Wolinowska, Marta Nieckarz, Pawel Wawrzyniak, Piotr Kieryl, Andrzej Plucienniczak, and Dariusz Bartosik Copyright © 2015 Marcin Adamczuk et al. All rights reserved. Nontuberculous Mycobacteria: Epidemiologic, Mycobacteriologic, and Clinical Aspects Tue, 16 Jun 2015 12:40:12 +0000 Mehdi Mirsaeidi, Parissa Farnia, Ruxana Sadikot, Po-Ren Hsueh, and Stefano Aliberti Copyright © 2015 Mehdi Mirsaeidi et al. All rights reserved. Isolation of Mycobacterium avium from Potable Water in Homes and Institutions of Patients with HIV Infection in Finland and the United States Tue, 09 Jun 2015 08:16:55 +0000 Symptomatic disease by nontuberculous mycobacteria has been linked to potable water from institutional and domestic potable water systems. Potable water samples were collected from homes and institutions of patients with AIDS. Colonization of potable water with nontuberculous mycobacteria was demonstrated in 230 (15%) of 1489 samples collected from domestic and institutional water systems of patients with HIV infection in the United States and Finland. Mycobacterium avium was the most common species and colonization was favored at temperatures of 40–50°C in recirculating hot water systems. Such systems are a plausible source of human infection and disease. Matti Ristola, Robert D. Arbeit, C. Fordham von Reyn, and C. Robert Horsburgh Jr. Copyright © 2015 Matti Ristola et al. All rights reserved. Specific Proteins in Nontuberculous Mycobacteria: New Potential Tools Thu, 28 May 2015 14:15:27 +0000 Nontuberculous mycobacteria (NTM) have been isolated from water, soil, air, food, protozoa, plants, animals, and humans. Although most NTM are saprophytes, approximately one-third of NTM have been associated with human diseases. In this study, we did a comparative proteomic analysis among five NTM strains isolated from several sources. There were different numbers of protein spots from M. gordonae (1,264), M. nonchromogenicum type I (894), M. nonchromogenicum type II (935), M. peregrinum (806), and M. scrofulaceum/Mycobacterium mantenii (1,486) strains, respectively. We identified 141 proteins common to all strains and specific proteins to each NTM strain. A total of 23 proteins were selected for its identification. Two of the common proteins identified (short-chain dehydrogenase/reductase SDR and diguanylate cyclase) did not align with M. tuberculosis complex protein sequences, which suggest that these proteins are found only in the NTM strains. Some of the proteins identified as common to all strains can be used as markers of NTM exposure and for the development of new diagnostic tools. Additionally, the specific proteins to NTM strains identified may represent potential candidates for the diagnosis of diseases caused by these mycobacteria. Patricia Orduña, Antonia I. Castillo-Rodal, Martha E. Mercado, Samuel Ponce de León, and Yolanda López-Vidal Copyright © 2015 Patricia Orduña et al. All rights reserved. Beninese Medicinal Plants as a Source of Antimycobacterial Agents: Bioguided Fractionation and In Vitro Activity of Alkaloids Isolated from Holarrhena floribunda Used in Traditional Treatment of Buruli Ulcer Thu, 28 May 2015 13:43:00 +0000 Buruli ulcer (BU) imposes a serious economic burden on affected households and on health systems that are involved in diagnosing the disease and treating patients. Research is needed to find cost-effective therapies for this costly disease. Plants have always been an important source of new pharmacologically active molecules. Consequently we decided to undertake the study of plants used in traditional treatment of BU in Benin and investigate their antimycobacterial activity as well as their chemical composition. Extracts from forty-four (44) plant species were selected on account of reported traditional uses for the treatment of BU in Benin and were assayed for antimycobacterial activities. Crude hydroethanolic extract from aerial parts of Holarrhena floribunda (G. Don) T. Durand and Schinz was found to have significant antimycobacterial activity against M. ulcerans (MIC = 125 µg/mL). We describe here the identification of four steroidal alkaloids from Mycobacterium ulcerans growth-inhibiting fractions of the alkaloidal extract of the aerial parts of Holarrhena floribunda. Holadysamine was purified in sufficient amount to allow the determination of its MCI (=50 µg/mL). These results give some support to the use of this plant in traditional medicine. Achille Yemoa, Joachim Gbenou, Dissou Affolabi, Mansourou Moudachirou, André Bigot, Séverin Anagonou, Françoise Portaels, Anandi Martin, and Joëlle Quetin-Leclercq Copyright © 2015 Achille Yemoa et al. All rights reserved. Sliding Motility, Biofilm Formation, and Glycopeptidolipid Production in Mycobacterium colombiense Strains Thu, 28 May 2015 12:29:28 +0000 Mycobacterium colombiense is a novel member of the Mycobacterium avium complex, which produces respiratory and disseminated infections in immunosuppressed patients. Currently, the morphological and genetic bases underlying the phenotypic features of M. colombiense strains remain unknown. In the present study, we demonstrated that M. colombiense strains displaying smooth morphology show increased biofilm formation on hydrophobic surfaces and sliding on motility plates. Thin-layer chromatography experiments showed that M. colombiense strains displaying smooth colonies produce large amounts of glycolipids with a chromatographic behaviour similar to that of the glycopeptidolipids (GPLs) of M. avium. Conversely, we observed a natural rough variant of M. colombiense (57B strain) lacking pigmentation and exhibiting impaired sliding, biofilm formation, and GPL production. Bioinformatics analyses revealed a gene cluster that is likely involved in GPL biosynthesis in M. colombiense CECT 3035. RT-qPCR experiments showed that motile culture conditions activate the transcription of genes possibly involved in key enzymatic activities of GPL biosynthesis. Milena Maya-Hoyos, John Leguizamón, Leonardo Mariño-Ramírez, and Carlos Y. Soto Copyright © 2015 Milena Maya-Hoyos et al. All rights reserved. Lung and Nodal Involvement in Nontuberculous Mycobacterial Disease: PET/CT Role Thu, 28 May 2015 09:31:39 +0000 Introduction. Systematic use of 18F-FDG PET/CT has the potential to simultaneously assess both pulmonary and lymph node involvement in nontuberculous mycobacterial (NTM) lung infection. Objective. The aim of the study was to evaluate the role of 18F-FDG PET/CT in the assessment of both mediastinal lymph nodes and lung involvement in NTM patients compared with active tuberculosis (TB) patients. Methods. 26 patients with pulmonary NTM disease were selected; six consecutive patients had undergone 18F-FDG PET/CT and data was compared with 6 active TB patients. Results. NTM exhibited different radiological lung patterns with an average SUV max value at PET/CT scan of 3,59 ± 2,32 (range 1,14 to 9,01) on pulmonary lesions and a mean value of SUV max 1,21 ± 0,29 (range 0,90 to 1,70) on mediastinal lymph nodes. Pulmonary lesions in TB showed an average SUV max value of 10,07 ± 6,45 (range 1,20 to 22,75) whilst involved mediastinal lymph nodes exhibited a mean SUV max value of 7,23 ± 3,03 (range 1,78 to 15,72). Conclusions. The differences in PET uptake in a broad range of lung lesions and lymph nodes between NTM and M. tuberculosis patients suggest a potential role for PET/CT scan in the diagnosis and management of pulmonary mycobacterial disease. Ginevra Del Giudice, Andrea Bianco, Antonio Cennamo, Giulia Santoro, Marco Bifulco, Carlo Marzo, and Gennaro Mazzarella Copyright © 2015 Ginevra Del Giudice et al. All rights reserved. Clinical Relevance of Nontuberculous Mycobacteria Isolated from Sputum in a Gold Mining Workforce in South Africa: An Observational, Clinical Study Thu, 28 May 2015 08:51:29 +0000 Background. The clinical relevance of nontuberculous mycobacteria (NTM), detected by liquid more than solid culture in sputum specimens from a South African mining workforce, is uncertain. We aimed to describe the current spectrum and relevance of NTM in this population. Methods. An observational study including individuals with sputum NTM isolates, recruited at workforce tuberculosis screening and routine clinics. Symptom questionnaires were administered at the time of sputum collection and clinical records and chest radiographs reviewed retrospectively. Results. Of 232 individuals included (228 (98%) male, median age 44 years), M. gordonae (60 individuals), M. kansasii (50), and M. avium complex (MAC: 38) were the commonest species. Of 38 MAC isolates, only 2 (5.3%) were from smear-positive sputum specimens and 30/38 grew in liquid but not solid culture. MAC was especially prevalent among symptomatic, HIV-positive individuals. HIV prevalence was high: 57/74 (77%) among those tested. No differences were found in probability of death or medical separation by NTM species. Conclusions. M. gordonae, M. kansasii, and MAC were the commonest NTM among miners with suspected tuberculosis, with most MAC from smear-negative specimens in liquid culture only. HIV testing and identification of key pathogenic NTM in this setting are essential to ensure optimal treatment. Clare L. van Halsema, Violet N. Chihota, Nicolaas C. Gey van Pittius, Katherine L. Fielding, James J. Lewis, Paul D. van Helden, Gavin J. Churchyard, and Alison D. Grant Copyright © 2015 Clare L. van Halsema et al. All rights reserved. Use of MALDI-TOF MS for Identification of Nontuberculous Mycobacterium Species Isolated from Clinical Specimens Thu, 28 May 2015 08:01:34 +0000 The aim of this study was to compare the results obtained for identification by MALDI-TOF of nontuberculous mycobacteria (NTM) isolated in clinical samples with those obtained by GenoType Mycobacterium CM/AS (common mycobacteria/additional species). A total of 66 Mycobacterium isolates from various clinical specimens (mainly respiratory) were tested in this study. They were identified using MALDI-TOF Bruker from strains isolated in Lowenstein, following the recommended protocol of heat inactivation and extraction, and were simultaneously analyzed through hybridization by GenoType Mycobacterium from liquid culture MGIT. Our results showed that identification by MALDI-TOF was correct in 98.4% (65/66) of NTM isolated in our clinical practice (M. avium, M. intracellulare, M. abscessus, M. chelonae, M. fortuitum, M. mucogenicum, M. kansasii, and M. scrofulaceum). MALDI-TOF was found to be an accurate, rapid, and cost-effective system for identification of mycobacteria species. María Concepción Mediavilla-Gradolph, Inmaculada De Toro-Peinado, María Pilar Bermúdez-Ruiz, María de los Ángeles García-Martínez, María Ortega-Torres, Natalia Montiel Quezel-Guerraz, and Begoña Palop-Borrás Copyright © 2015 María Concepción Mediavilla-Gradolph et al. All rights reserved. Nontuberculous Mycobacteria Isolation from Clinical and Environmental Samples in Iran: Twenty Years of Surveillance Thu, 28 May 2015 06:34:50 +0000 Nontuberculous mycobacteria (NTM) are opportunistic pathogens that are widely distributed in the environment. There is a lack of data on species distribution of these organisms from Iran. This study consists of a review of NTM articles published in Iran between the years 1992 and 2014. In this review, 20 articles and 14 case reports were identified. Among the 20 articles, 13 (65%) studies focused on NTM isolates from clinical specimens, 6 (30%) studies examined NTM isolates from environmental samples, and one (5%) article included both clinical and environmental isolates. M. fortuitum (229/997; 23%) was recorded as the most prevalent and rapid growing mycobacteria (RGM) species in both clinical (28%) and environmental (19%) isolated samples (P < 0.05). Among slow growing mycobacteria (SGM), M. simiae (103/494; 21%) demonstrated a higher frequency in clinical samples whereas in environmental samples it was M. flavescens (44/503; 9%). These data represent information from 14 provinces out of 31 provinces of Iran. No information is available in current published data on clinical or environmental NTM from the remaining 17 provinces in Iran. These results emphasize the potential importance of NTM as well as the underestimation of NTM frequency in Iran. NTM is an important clinical problem associated with significant morbidity and mortality in Iran. Continued research is needed from both clinical and environmental sources to help clinicians and researchers better understand and address NTM treatment and prevention. Ali Akbar Velayati, Parissa Farnia, Mohadese Mozafari, and Mehdi Mirsaeidi Copyright © 2015 Ali Akbar Velayati et al. All rights reserved. Species Identification and Clarithromycin Susceptibility Testing of 278 Clinical Nontuberculosis Mycobacteria Isolates Thu, 28 May 2015 06:12:04 +0000 Purpose of this paper is to analyze different species’ proportion of nontuberculosis mycobacteria (NTM) and susceptibility to clarithromycin of different species. 278 clinical NTM isolates were identified into species by using 16S rRNA, rpoB and hsp65. Then clarithromycin susceptibility testing against different species was done separately, using microplate Alamar Blue assay. Finally, resistance isolates’ erm(41) of M. abscessus were sequenced in order to analyze mechanisms for clarithromycin resistant. In this test, 131 isolates (47%) belonged to M. avium complex (MAC), and 70 isolates (25%) belonged to M. abscessus. Nearly all the M. abscessus subsp. abscessus resistant to clarithromycin had T28 in erm(41). However, all the M. abscessus subsp. abscessus susceptible to clarithromycin had C28 in erm(41). In this study, we find that MAC was the most common pathogens of NTM, and the second one was M. abscessus. However, M. chelonei, M. fuerth, and M. gordon were rare. Clarithromycin had a good inhibition activity against all the NTM species except M. abscessus subsp. abscessus. The erm(41) genotype is of high relevance to clarithromycin resistance. Wenjuan Nie, Hongfei Duan, Hairong Huang, Yu Lu, and Naihui Chu Copyright © 2015 Wenjuan Nie et al. All rights reserved. Nontuberculous Mycobacteria in Noncystic Fibrosis Bronchiectasis Wed, 27 May 2015 14:21:27 +0000 During the past decades, a growing interest has been raised in evaluating nontuberculous mycobacteria (NTM) in patients with noncystic fibrosis bronchiectasis (NCFBE). This paper reviews several aspects of the correlations between NTM and NCFBE, including pathogenesis, radiological features, diagnosis, and management. Bronchiectasis and NTM lung disease are connected, but which one comes first is still an unresolved question. The rate of NTM lung disease in NCFBE varies through the studies, from 5% to 30%. The most frequent species isolated is MAC. NCFBE patients affected by NTM infection frequently present coinfections, including both other different NTM species and microorganisms, such as P. aeruginosa. Once a diagnosis of NTM disease has been reached, the initiation of therapy is not always mandatory. NTM species isolated, patients’ conditions, and disease severity and its evolution should be considered. Risk factors for disease progression in NCFBE patients with NTM are low body mass index, cavitary disease, consolidations, and macrolide resistance at presentation. Giulia Bonaiti, Alberto Pesci, Almerico Marruchella, Giuseppe Lapadula, Andrea Gori, and Stefano Aliberti Copyright © 2015 Giulia Bonaiti et al. All rights reserved. Nontuberculous Mycobacterial Ocular Infections: A Systematic Review of the Literature Wed, 27 May 2015 14:16:54 +0000 Nontuberculous or atypical mycobacterial ocular infections have been increasing in prevalence over the past few decades. They are known to cause periocular, adnexal, ocular surface and intraocular infections and are often recalcitrant to medical therapy. These infections can potentially cause detrimental outcomes, in part due to a delay in diagnosis. We review 174 case reports and series on nontuberculous mycobacterial (NTM) ocular infections and discuss etiology, microbiology, risk factors, diagnosis, clinical presentation, and treatment of these infections. History of interventions, trauma, foreign bodies, implants, contact lenses, and steroids are linked to NTM ocular infections. Steroid use may prolong the duration of the infection and cause poorer visual outcomes. Early diagnosis and initiation of treatment with multiple antibiotics are necessary to achieve the best visual outcome. Wajiha J. Kheir, Huda Sheheitli, Maamoun Abdul Fattah, and Rola N. Hamam Copyright © 2015 Wajiha J. Kheir et al. All rights reserved. Insufficient Discriminatory Power of Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry Dendrograms to Determine the Clonality of Multi-Drug-Resistant Acinetobacter baumannii Isolates from an Intensive Care Unit Mon, 25 May 2015 09:23:11 +0000 While pulsed-field gel electrophoresis (PFGE) is recognized as the gold standard method for clonality analysis, MALDI-TOF MS has recently been spotlighted as an alternative tool for species identification. Herein, we compared the dendrograms of multi-drug-resistant (MDR) Acinetobacter baumannii isolates by using MALDI-TOF MS with those by using PFGE. We used direct colony and protein extraction methods for MALDI-TOF MS dendrograms. The isolates with identical PFGE patterns were grouped into different branches in MALDI-TOF MS dendrograms. Among the isolates that were classified as very close isolates in MALDI-TOF MS dendrogram, PFGE band patterns visually showed complete differences. We numeralized similarity among isolates by measuring distance levels. The Spearman rank correlation coefficient values were 0.449 and 0.297 between MALDI-TOF MS dendrogram using direct colony and protein extraction method versus PFGE, respectively. This study is the first paper focusing solely on the dendrogram function of MALDI-TOF MS compared with PFGE. Although MALDI-TOF MS is a promising tool to identify species in a rapid manner, our results showed that MALDI-TOF MS dendrograms could not substitute PFGE for MDR Acinetobacter baumannii clonality analysis. John Hoon Rim, Yangsoon Lee, Sung Kuk Hong, Yongjung Park, MyungSook Kim, Roshan D’Souza, Eun Suk Park, Dongeun Yong, and Kyungwon Lee Copyright © 2015 John Hoon Rim et al. All rights reserved. A Novel Method for the Synthesis of -Ofloxacin Kits Using D-Penicillamine as Coligand and Their Application as Infection Imaging Agent Tue, 19 May 2015 14:13:49 +0000 The employment of radiopharmaceuticals is increasing nowadays for infection imaging and early execution of patients having infectious or inflammatory complaints. The main aim of this study was to discover a novel method for the labeling of ofloxacin with , optimization of labelling conditions to get higher percent yield, to assess kits radiochemical purity, in vitro stability, partition coefficient, protein binding, and intracellular accumulation in Pseudomonas aeruginosa, Salmonella typhi, and Escherichia coli in infected rabbits. Maximum labeling efficiency was achieved when 1.5 mg ofloxacin was labeled with 10–20 mCi sodium pertechnetate in the presence of 3 mg D-penicillamine, 75 μg SnCl2. In vitro binding and biodistribution in Pseudomonas aeruginosa, Salmonella typhi, and Escherichia coli showed good results. This new complex is efficient for the imaging of infections caused by Gram-positive and Gram-negative bacteria. Muhammad Abdul Qadir, Shabnam Shahzad, Rashid Rasheed, Mahmood Ahmed, Shahzad Anwar, and Syeda Kiran Shahzadi Copyright © 2015 Muhammad Abdul Qadir et al. All rights reserved. Risk Factors and Scoring System for Predicting Bacterial Resistance to Cefepime as Used Empirically in Haematology Wards Tue, 05 May 2015 14:29:57 +0000 Objectives. Bacterial resistance is of growing concern in haematology wards. As the inappropriate administration of empirical antibacterial may alter survival, we studied risk factors for resistance to our usual empirical first-line antibacterial therapy, cefepime. Methods. We retrospectively studied 103 first episodes of bacteraemia recorded in our haematology department over 2.5 years. Risk factors for cefepime-resistance were identified by multivariate logistic regression with backward selection (). A scoring system for predicting cefepime-resistance was built on independent factor, with an internal validation by the bootstrap resampling technique. Results. 38 (37%) episodes were due to Gram-negative bacteria. Fifty (49%) were due to bacteria resistant to cefepime. Cefepime resistance was significantly associated with a decreased survival at day 30 (). Three risk factors were independently associated with cefepime-resistance: acute lymphoblastic leukaemia; ≥18 days since hospital admission; and receipt of any -lactam in the last month. Patients with ≥2 of these risk factors had a probability of 86% (CI 95%, 25 to 100%) to carry a cefepime-resistant strain. Conclusion. Using our scoring system should reduce the indication of very broad antibacterial regimens in the empirical, first-line treatment of febrile hematology patients in more than 80% of the cases. Hicham El Maaroufi, Agathe Goubard, Rabah Redjoul, Patrick Legrand, Cécile Pautas, Mohamed Mikdame, Kamal Doghmi, Andréa Toma, Sébastien Maury, Michael Schwarzinger, and Catherine Cordonnier Copyright © 2015 Hicham El Maaroufi et al. All rights reserved. Dealing with Stigma: Experiences of Persons Affected by Disabilities and Leprosy Wed, 15 Apr 2015 13:19:34 +0000 Persons affected by leprosy or by disabilities face forms of stigma that have an impact on their lives. This study seeks to establish whether their experiences of stigma are similar, with a view to enabling the two groups of people to learn from each other. Accounts of experiences of the impact of stigma were obtained using in-depth interviews and focus group discussion with people affected by leprosy and by disabilities not related to leprosy. The analysis shows that there are a lot of similarities in impact of stigma in terms of emotions, thoughts, behaviour, and relationships between the two groups. The main difference is that those affected by leprosy tended to frame their situation in medical terms, while those living with disabilities described their situation from a more social perspective. In conclusion, the similarities offer opportunities for interventions and the positive attitudes and behaviours can be modelled in the sense that both groups can learn and benefit. Research that tackles different aspects of stigmatization faced by both groups could lead to inclusive initiatives that help individuals to come to terms with the stigma and to advocate against exclusion and discrimination. Mimi Lusli, Marjolein B. M. Zweekhorst, Beatriz Miranda-Galarza, Ruth M. H. Peters, Sarah Cummings, Francisia S. S. E. Seda, Joske F. G. Bunders, and Irwanto Copyright © 2015 Mimi Lusli et al. All rights reserved. Educational Effectiveness, Target, and Content for Prudent Antibiotic Use Sun, 05 Apr 2015 11:07:29 +0000 Widespread antimicrobial use and concomitant resistance have led to a significant threat to public health. Because inappropriate use and overuse of antibiotics based on insufficient knowledge are one of the major drivers of antibiotic resistance, education about prudent antibiotic use aimed at both the prescribers and the public is important. This review investigates recent studies on the effect of interventions for promoting prudent antibiotics prescribing. Up to now, most educational efforts have been targeted to medical professionals, and many studies showed that these educational efforts are significantly effective in reducing antibiotic prescribing. Recently, the development of educational programs to reduce antibiotic use is expanding into other groups, such as the adult public and children. The investigation of the contents of educational programs for prescribers and the public demonstrates that it is important to develop effective educational programs suitable for each group. In particular, it seems now to be crucial to develop appropriate curricula for teaching medical and nonmedical (pharmacy, dentistry, nursing, veterinary medicine, and midwifery) undergraduate students about general medicine, microbial virulence, mechanism of antibiotic resistance, and judicious antibiotic prescribing. Chang-Ro Lee, Jung Hun Lee, Lin-Woo Kang, Byeong Chul Jeong, and Sang Hee Lee Copyright © 2015 Chang-Ro Lee et al. All rights reserved. Prevalence of Nontuberculous Mycobacteria among Extrapulmonary Tuberculosis Cases in Tertiary Care Centers in Northern India Thu, 26 Mar 2015 09:00:12 +0000 The reports of nontuberculous mycobacteria (NTM) associated with extrapulmonary diseases are increasing in tertiary care hospitals. Despite a significant increase in knowledge about NTM infections, they still represent a diagnostic and therapeutic challenge. The aim of this study is to know the prevalence of NTN among extrapulmonary tuberculosis cases in tertiary care centers in Northern India. A total of 227 culture positive isolates from 756 cases were tested for niacin production and catalase assay. BIO-LINE SD Ag MPT64 TB test and final identification and differentiation between MTBC and different species of NTM were further confirmed by GenoType Mycobacterium CM/AS assay. 71 cases (9.3%) were positive for AFB by ZN staining and 227 cases (30.1%) were positive for mycobacteria by culture. Niacin production and catalase activity were negative in 62/227 (27.4%) strains and after using a panel of different biochemicals and final confirmation by GenoType Mycobacterium CM assay. Out of 227 cultures tested, 165 (72.6%) strains were confirmed as M. tuberculosis complex, and 62 (27.4%) were confirmed as NTM. The most common NTM species identified were M. fortuitum 17 (27.5%) and M. intracellulare 13 (20.9%). The rapid identification of NTM species may help in targeted therapy and management of the diseases. A. K. Maurya, V. L. Nag, S. Kant, R. A. S. Kushwaha, M. Kumar, A. K. Singh, and T. N. Dhole Copyright © 2015 A. K. Maurya et al. All rights reserved. The Epidemiology and Economic Burden of Clostridium difficile Infection in Korea Mon, 02 Mar 2015 11:19:05 +0000 The prevalence of Clostridium difficile infection and the associated burden have recently increased in many countries. While the main risk factors for C. difficile infection include old age and antibiotic use, the prevalence of this infection is increasing in low-risk groups. These trends highlight the need for research on C. difficile infection. This study pointed out the prevalence and economic burden of C. difficile infection and uses the representative national data which is primarily from the database of the Korean Health Insurance Review and Assessment Service, for 2008–2011. The annual economic cost was measured using a prevalence approach, which sums the costs incurred to treat C. difficile infection. C. difficile infection prevalence was estimated to have increased from 1.43 per 100,000 in 2008 to 5.06 per 100,000 in 2011. Moreover, mortality increased from 69 cases in 2008 to 172 in 2011. The economic cost increased concurrently, from $2.4 million in 2008 to $7.6 million, $10.5 million, and $15.8 million in 2009, 2010, and 2011, respectively. The increasing economic burden of C. difficile infection over the course of the study period emphasizes the need for intervention to minimize the burden of a preventable illness like C. difficile infection. Hyung-Yun Choi, So-Youn Park, Young-Ae Kim, Tai-Young Yoon, Joong-Myung Choi, Bong-Keun Choe, So-Hee Ahn, Seok-Jun Yoon, Ye-Rin Lee, and In-Hwan Oh Copyright © 2015 Hyung-Yun Choi et al. All rights reserved. HIV-Associated Neurocognitive Disorders: The Relationship of HIV Infection with Physical and Social Comorbidities Sun, 01 Mar 2015 12:12:45 +0000 The prevalence of HIV (human immunodeficiency virus) associated neurocognitive disorders (HAND) will undoubtedly increase with the improved longevity of HIV-infected persons. HIV infection, itself, as well as multiple physiologic and psychosocial factors can contribute to cognitive impairment and neurologic complications. These comorbidities confound the diagnosis, assessment, and interventions for neurocognitive disorders. In this review, we discuss the role of several key comorbid factors that may contribute significantly to the development and progression of HIV-related neurocognitive impairment, as well as the current status of diagnostic strategies aimed at identifying HIV-infected individuals with impaired cognition and future research priorities and challenges. Ellen M. Tedaldi, Nancy L. Minniti, and Tracy Fischer Copyright © 2015 Ellen M. Tedaldi et al. All rights reserved. Analysis of Monocytic and Granulocytic Myeloid-Derived Suppressor Cells Subsets in Patients with Hepatitis C Virus Infection and Their Clinical Significance Sun, 01 Mar 2015 11:17:57 +0000 Myeloid-derived suppressor cells (MDSCs) have been shown to inhibit T-cell responses in many diseases, but, in hepatitis C virus (HCV) infected patients, MDSCs are still poorly studied. In this assay, we investigated the phenotype and frequency of two new populations of MDSCs denoted as monocytic and granulocytic MDSCs (M-MDSCs and G-MDSCs) in HCV infected patients and analyzed their clinical significance in these patients respectively. We found that the frequency of CD14+ cells (M-MDSCs) from HCV infected patients (mean ± SE, 3.134% ± 0.340%) was significantly increased when compared to healthy controls (mean ± SE, 1.764% ± 0.461%) (Z = −2.438, P = 0.015), while there was no statistical difference between the frequency of CD33+CD11b+CD15+ (G-MDSCs) of HCV infected patients and healthy donors (0.201% ± 0.038% versus 0.096% ± 0.026%, P > 0.05), which suggested that HCV infection could cause the proliferation of M-MDSCs instead of G-MDSCs. Besides, we found that the frequency of M-MDSCs in HCV infected patients had certain relevance with age (r = 0.358, P = 0.003); patients older than 40 years old group (mean ± SE, 3.673% ± 0.456%) had a significantly higher frequency of M-MDSCs than that of age less than 40 years old group (mean ± SE, 2.363% ± 0.482%) (Z = −2.685, P = 0.007). The frequency of M-MDSCs, however, had no correlation with HCV RNA loads, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and the level of liver inflammation degree. Gang Ning, Lanhui She, Lirong Lu, Ying Liu, Yingfu Zeng, Ying Yan, and Chaoshuang Lin Copyright © 2015 Gang Ning et al. All rights reserved. Clinical Usefulness of the 2010 Clinical and Laboratory Standards Institute Revised Breakpoints for Cephalosporin Use in the Treatment of Bacteremia Caused by Escherichia coli or Klebsiella spp. Sun, 22 Feb 2015 06:52:46 +0000 We investigated the clinical usefulness of the revised 2010 Clinical and Laboratory Standards Institute (CLSI) breakpoints for Escherichia coli and Klebsiella spp. Of 2,623 patients with bacteremia caused by E. coli or Klebsiella spp., 573 who had been treated appropriately with cephalosporin based on the CLSI 2009 guidelines were enrolled. There were no differences in the rates of treatment failure or mortality between the appropriately and inappropriately treated groups according to the CLSI 2010 guidelines. Additionally, in the matched case-control analysis, the treatment failure rate was higher in bacteremic patients with extended-spectrum β-lactamase- (ESBL-) producing but cephalosporin-susceptible organisms than in those with ESBL-nonproducing isolates when patients with urinary tract infections were excluded (44% and 0%, resp., ). In patients with bacteremia caused by E. coli or Klebsiella spp., the revised CLSI 2010 guidelines did not lead to poorer outcomes. However, ESBL production appeared to be associated with poor clinical outcomes in patients with bacteremia from sources other than the urinary tract. Nam Su Ku, Hae-Sun Chung, Jun Yong Choi, Dongeun Yong, Kyungwon Lee, June Myung Kim, and Yunsop Chong Copyright © 2015 Nam Su Ku et al. All rights reserved. Phytochemical Analysis and Modulation of Antibiotic Activity by Luehea paniculata Mart. & Zucc. (Malvaceae) in Multiresistant Clinical Isolates of Candida Spp. Mon, 16 Feb 2015 09:17:47 +0000 The high incidence of fungal infections has led to the continuous search for new drugs. Extracts of Luehea paniculata, a tree of multiple medicinal uses, were evaluated for anti-Candida activity, as well as its modulator potential of the Fluconazole antibiotic. Chemical prospecting of ethanol extracts of leaf and bark was carried out, the quantification of total phenols and flavonoids, characterized by the HPLC-DAD technique. The rosmarinic acid and the vitexin flavonoid were observed as major constituents in ELELP and ESWELP, respectively. Antioxidant activity was also evaluated by the method of scavenging the free radical DPPH, and quercetin was used as standard, obtaining IC50 values: 0.341 (mg/mL) for ELELP and 0.235 (mg/mL) for ESWELP. The microdilution assay was performed for antifungal activity against strains of Candida albicans, C. krusei, and C. tropicalis and showed minimum inhibitory concentrations values ≥1024 μg/mL. In the modulator action of extracts on Fluconazole against multiresistant clinical isolates of Candida (subinhibitory concentration minimum of 128 μg/mL), a significant synergism was observed, indicating that the extracts potentiated the antifungal effect against C. tropicalis, where antioxidant flavonoids could be responsible. This is the first report about modifying activity of the antibiotic action of a species of the genus Luehea. João T. Calixto Júnior, Selene M. Morais, Clécio G. Martins, Larissa G. Vieira, Maria Flaviana B. Morais-Braga, Joara N. P. Carneiro, Antonio J. P. Machado, Irwin R. A. Menezes, Saulo R. Tintino, and Henrique D. M. Coutinho Copyright © 2015 João T. Calixto Júnior et al. All rights reserved. Predictive Contribution of Neutrophil/Lymphocyte Ratio in Diagnosis of Brucellosis Tue, 03 Feb 2015 08:46:59 +0000 Here we wanted to investigate predictive value of neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) in the diagnosis of brucellosis. Thirty-two brucellosis patients diagnosed with positive serum agglutination test and thirty-two randomized healthy subjects were enrolled in this study retrospectively. Result with ROC analyzes the baseline NLR and hemoglobin values were found to be significantly associated with brucellosis (, , resp.). Herein we demonstrated for the first time that NLR values were significantly associated with brucellosis. This situation can help clinicians during diagnosis of brucellosis. Serdar Olt, Hasan Ergenç, and Seyyid Bilal Açıkgöz Copyright © 2015 Serdar Olt et al. All rights reserved. Research on Bacterial Virulence in the Developing Countries Mon, 02 Feb 2015 07:21:20 +0000 Angel Cataldi, Ana Lucia Nascimento, Armando Acosta, and Chensong Wan Copyright © 2015 Angel Cataldi et al. All rights reserved. Latent Tuberculosis Infection among a Large Cohort of Medical Students at a Teaching Hospital in Italy Sun, 01 Feb 2015 11:52:46 +0000 The surveillance of latent tuberculosis infection (LTBI) in both healthcare workers and healthcare students is considered fundamental for tuberculosis (TB) prevention. The aim of the present study was to estimate LTBI prevalence and evaluate potential risk-factors associated with this condition in a large cohort of medical students in Italy. In a cross-sectional study, performed between March and December 2012, 1511 eligible subjects attending the Medical School of the University of Genoa, trained at the IRCCS San Martino-IST Teaching Hospital of Genoa, were actively called to undergo the tuberculin skin test (TST). All the TST positive cases were confirmed with an interferon-gamma release assay (IGRA). A standardized questionnaire was collected for multivariate risk analysis. A total of 1302 (86.2%) students underwent TST testing and completed the questionnaire. Eleven subjects (0.8%) resulted TST positive and LTBI diagnosis was confirmed in 2 (0.1%) cases. Professional exposure to active TB patients (OR 21.7, 95% CI 2.9–160.2; value 0.003) and previous BCG immunization (OR 28.3, 95% CI 3.0–265.1; value 0.003) are independently associated with TST positivity. Despite the low prevalence of LTBI among Italian medical students, an occupational risk of TB infection still exists in countries with low circulation of Mycobacterium tuberculosis. Paolo Durando, Cristiano Alicino, Andrea Orsi, Ilaria Barberis, Chiara Paganino, Guglielmo Dini, Giovanni Mazzarello, Valerio Del Bono, Claudio Viscoli, Francesco Copello, Dimitri Sossai, Giovanni Orengo, Laura Sticchi, Filippo Ansaldi, and Giancarlo Icardi Copyright © 2015 Paolo Durando et al. All rights reserved. WHO Multidrug Therapy for Leprosy: Epidemiology of Default in Treatment in Agra District, Uttar Pradesh, India Sun, 01 Feb 2015 11:30:07 +0000 Aim. To study the magnitude of default, time of default, its causes, and final clinical outcome. Methods. Data collected in active surveys in Agra is analyzed. Patients were given treatment after medical confirmation and were followed up. The treatment default and other clinical outcomes were recorded. Results. Patients who defaulted have comparable demographic characteristics. However, among defaulters more women (62.7% in PB, 42.6% in MB) were seen than those in treatment completers (PB 52.7% and MB 35.9%). Nerve involvement was high in treatment completers: 45.7% in PB and 91.3% in MB leprosy. Overall default rate was lower (14.8%) in ROM than (28.8%) in standard MDT for PB leprosy (, ) and also for MB leprosy: 9.1% in ROM compared to 34.5% in MDT (, ). Default rate was not different (28.8% versus 34.5%, ) in both types of leprosy given MDT. Most patients defaulted at early stage of treatment and mainly due to manageable side effects. Conclusion. The default in standard MDT both for PB and MB leprosy was observed to be significantly higher than in ROM treatment. Most defaults occurred at early stage of treatment and major contribution of default is due to side effects like drowsiness, weakness, vomiting, diarrhea, and so forth, related to poor general health. Although about half of the defaulters were observed to be cured 2.2% in PB-MDT and 10.9% of MB-MDT developed disability. This is an issue due to default. Attempts are needed to increase treatment compliance. The use of specially designed disease related health education along with easily administered drug regimens may help to reduce default. Anil Kumar, Anita Girdhar, Joy Kumar Chakma, and Bhuwneswar Kumar Girdhar Copyright © 2015 Anil Kumar et al. All rights reserved. Impairment of Neutrophil Migration to Remote Inflammatory Site during Lung Histoplasmosis Thu, 29 Jan 2015 14:24:10 +0000 Histoplasma capsulatum (Hc) induces a pulmonary disease in which leukotrienes promote activation and recruitment of effectors cells. It is also well-recognized that leukotriene B4 (LTB4) and platelet-activating factor (PAF) induce leukocyte recruitment to inflammatory sites. We investigated the impact of pulmonary Hc infection on PMN migration to a remote inflammatory site. Our results show that pulmonary Hc infection impairs LTB4- or PAF-stimulated PMN recruitment to air pouch. Yet, remote inflammation did not modify PMN numbers in the bronchoalveolar lavage fluid (BALF) of Hc-infected mice. Interestingly, the concomitant administration of PAF and LTB4 receptor antagonists inhibited PMN recruitment to both BALF and the remote site, demonstrating cooperation between both mediators. Along that line, our results show that PAF-elicited PMN chemotaxis was abrogated in 5-lipoxygenase-deficient animals. These results suggest caution in the indiscriminate use of anti-inflammatory drugs during infectious diseases. Alexandra I. Medeiros, Adriana Secatto, Caroline Bélanger, Carlos A. Sorgi, Pierre Borgeat, Sylvie Marleau, and Lúcia H. Faccioli Copyright © 2015 Alexandra I. Medeiros et al. All rights reserved. High In Vitro Antibacterial Activity of Pac-525 against Porphyromonas gingivalis Biofilms Cultured on Titanium Thu, 29 Jan 2015 13:35:36 +0000 In order to investigate the potential of short antimicrobial peptides (AMPs) as alternative antibacterial agents during the treatment of peri-implantitis, the cytotoxic activity of three short AMPs, that is, Pac-525, KSL-W, and KSL, was determined using the MTT assay. The antimicrobial activity of these AMPs, ranging in concentration from 0.0039 mg/mL to 0.5 mg/mL, against the predominant planktonic pathogens, including Streptococcus sanguis, Fusobacterium nucleatum, and Porphyromonas gingivalis, involved in peri-implantitis was investigated. Furthermore, 2-day-old P. gingivalis biofilms cultured on titanium surfaces were treated with Pac-525 and subsequently observed and analysed using confocal laser scanning microscopy (CLSM). The average cell proliferation curve indicated that there was no cytotoxicity due to the three short AMPs. The minimum inhibitory concentration and minimum bactericidal concentration values of Pac-525 were 0.0625 mg/mL and 0.125 mg/mL, respectively, for P. gingivalis and 0.0078 mg/mL and 0.0156 mg/mL, respectively, for F. nucleatum. Using CLSM, we confirmed that compared to 0.1% chlorhexidine, 0.5 mg/mL of Pac-525 caused a significant decrease in biofilm thickness and a decline in the percentage of live bacteria. These data indicate that Pac-525 has unique properties that might make it suitable for the inhibition the growth of pathogenic bacteria around dental implants. Ji-yin Li, Xue-jin Wang, Li-na Wang, Xiao-xia Ying, Xiang Ren, Hui-ying Liu, Li Xu, and Guo-wu Ma Copyright © 2015 Ji-yin Li et al. All rights reserved. Pan-Genome Analysis of Human Gastric Pathogen H. pylori: Comparative Genomics and Pathogenomics Approaches to Identify Regions Associated with Pathogenicity and Prediction of Potential Core Therapeutic Targets Thu, 29 Jan 2015 06:54:04 +0000 Helicobacter pylori is a human gastric pathogen implicated as the major cause of peptic ulcer and second leading cause of gastric cancer (~70%) around the world. Conversely, an increased resistance to antibiotics and hindrances in the development of vaccines against H. pylori are observed. Pan-genome analyses of the global representative H. pylori isolates consisting of 39 complete genomes are presented in this paper. Phylogenetic analyses have revealed close relationships among geographically diverse strains of H. pylori. The conservation among these genomes was further analyzed by pan-genome approach; the predicted conserved gene families (1,193) constitute ~77% of the average H. pylori genome and 45% of the global gene repertoire of the species. Reverse vaccinology strategies have been adopted to identify and narrow down the potential core-immunogenic candidates. Total of 28 nonhost homolog proteins were characterized as universal therapeutic targets against H. pylori based on their functional annotation and protein-protein interaction. Finally, pathogenomics and genome plasticity analysis revealed 3 highly conserved and 2 highly variable putative pathogenicity islands in all of the H. pylori genomes been analyzed. Amjad Ali, Anam Naz, Siomar C Soares, Marriam Bakhtiar, Sandeep Tiwari, Syed S Hassan, Fazal Hanan, Rommel Ramos, Ulisses Pereira, Debmalya Barh, Henrique César Pereira Figueiredo, David W. Ussery, Anderson Miyoshi, Artur Silva, and Vasco Azevedo Copyright © 2015 Amjad Ali et al. All rights reserved. Oxidative Stress Control by Apicomplexan Parasites Wed, 28 Jan 2015 09:44:33 +0000 Apicomplexan parasites cause infectious diseases that are either a severe public health problem or an economic burden. In this paper we will shed light on how oxidative stress can influence the host-pathogen relationship by focusing on three major diseases: babesiosis, coccidiosis, and toxoplasmosis. Soraya S. Bosch, Thales Kronenberger, Kamila A. Meissner, Flávia M. Zimbres, Dirk Stegehake, Natália M. Izui, Isolmar Schettert, Eva Liebau, and Carsten Wrenger Copyright © 2015 Soraya S. Bosch et al. All rights reserved. Investigation of Ser315 Substitutions within katG Gene in Isoniazid-Resistant Clinical Isolates of Mycobacterium tuberculosis from South India Wed, 28 Jan 2015 09:44:30 +0000 Mutation at codon 315 of katG gene is the major cause for isoniazid (INH) resistance in Mycobacterium tuberculosis (M. tuberculosis). Substitution at codon 315 of katG gene was analyzed in 85 phenotypically resistant isolates collected from various parts of southern India by direct sequencing method. The obtained results were interpreted in the context of minimum inhibitory concentration (MIC) of INH. Of the 85 phenotypically resistant isolates, 56 (66%) were also correlated by the presence of resistance mutations in the katG gene; 47 of these isolates had ACC, 6 had AAC, 2 had ATC, and one had CGC codon. The frequency of Ser315 substitution in katG gene was found to be higher (70%) amongst multidrug-resistant (MDR) strains than among non-MDR (61%) INH-resistant isolates. Further, the frequency of mutations was found to be greater (74%) in isolates with higher MIC values in contrast to those isolates with low MIC values (58%). Therefore, the study identified high prevalence of Ser315Thr substitution in katG gene of INH-resistant isolates from south India. Also, isolates harboring this substitution were found to be associated with multidrug and high level INH resistance. A. Nusrath Unissa, N. Selvakumar, Sujatha Narayanan, C. Suganthi, and L. E. Hanna Copyright © 2015 A. Nusrath Unissa et al. All rights reserved. Clinical Epidemiology and Paraclinical Findings in Tuberculosis Patients in North of Iran Wed, 28 Jan 2015 06:39:44 +0000 Background. Mycobacterium tuberculosis (M.TB) causes a wide spectrum of clinical diseases. The prevalence of TB is different in various parts of Iran and throughout the world. The present study aimed to determine the clinical epidemiology and paraclinical findings of TB. Methods. A cross-sectional study was conducted from 2008 to 2013. Patient demographic, clinical, and radiologic characteristics, picked up from the TB patient’s files, were collected using a standard questionnaire format. Data was entered and analyzed using the SPSS version 16 statistical software and P value < 0.05 was considered statistically significant. Results. Out of 212 patients enrolled in this study 62% were male and the mean age was about 50 years old. 98.6% were Iranian, and 46.2% were rural. Prevalence of smear-positive TB was 66.4%. Prevalence of positive PPD was 50.7% with no significant difference between HIV-positive and -negative patients (P = 0.8). Prevalence of diabetes mellitus was 17%. 36% of the patients had history of smoking and about 29.3% were addicted to narcotics. Cough was the most common symptom (94.5%) and 84% had sputum. 15 cases (7%) had extrapulmonary TB. The mean time between the onset of symptoms and admission was 46.5 days. The delay for admission between urban and rural populations was not significantly different (P = 0.68); but for those who were in prison, the delay was significant (P = 0.02). About 46% of the patients had cavitary lesions in CXRs. Conclusion. Timely diagnosis of TB especially in prisoners by understanding its most important epidemiologic characteristics and clinical features can help to make an early treatment and prevent spread of mycobacteria and their complications. Farhang Babamahmoodi, Ahmad Alikhani, Jamshid Yazdani Charati, Amir Ghovvati, Fatemeh Ahangarkani, Leila Delavarian, and Abdolreza Babamahmoodi Copyright © 2015 Farhang Babamahmoodi et al. All rights reserved. Development of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Trichosporon asahii in Experimental and Clinical Samples Tue, 27 Jan 2015 08:43:07 +0000 Invasive trichosporonosis is a deep mycosis found mainly in immunocompromised hosts, and the major pathogen is Trichosporon asahii. We detected the species-specific intergenic spacers (IGS) of rRNA gene of T. asahii using a loop-mediated isothermal amplification (LAMP) assay in 15 isolates with 3 different visualization methods, including SYBR green detection, gel electrophoresis, and turbidimetric methods. The LAMP assay displayed superior rapidity to other traditional methods in the detection time; that is, only 1 h was needed for detection and identification of the pathogen DNA. Furthermore, the detection limit of the LAMP assay was more sensitive than the PCR assay. We also successfully detect the presence of T. asahii in samples from experimentally infected mice and samples from patients with invasive trichosporonosis caused by T. asahii, suggesting that this method may become useful in clinical applications in the near future. Jianfeng Zhou, Yong Liao, Haitao Li, Xuelian Lu, Xiufeng Han, Yanli Tian, Shanshan Chen, and Rongya Yang Copyright © 2015 Jianfeng Zhou et al. All rights reserved. Clinical Significance of Neutrophil Gelatinase-Associated Lipocalin in Crimean-Congo Hemorrhagic Fever Thu, 22 Jan 2015 13:28:46 +0000 Neutrophil gelatinase-associated lipocalin (NGAL), which is an important prognostic marker for sepsis and inflammatory diseases, is mostly released from neutrophils. Crimean-Congo hemorrhagic fever (CCHF) patients are generally neutropenic. We aimed to investigate whether there is a change in serum NGAL level and to investigate its effect on the recovery time (RT) during the course of CCHF. A total of 40 CCHF patients (19 females and 21 males) and 34 healthy controls (17 females and 17 males) were included in the study. The serum NGAL level and biochemical and hematological parameters were checked. The NGAL level of CCHF patients was significantly higher than that of the healthy controls (). A multivariate analysis showed that the independent prognostic factor for the prediction of the RT is the NGAL level (odds ratio [OR] 0.3, 95% confidence interval [Cl] 0.1–0.4, ). An elevated NGAL level was found to be associated with an increased RT in CCHF patients. The NGAL levels of CHHF patients might be elevated due to increased cytokine release, the presence of a tissue injury, and the release of immature neutrophils from the bone marrow into the peripheral stream. This may be a good prognostic factor in CHHF patients. Ayse Erturk, Erkan Cure, Emine Parlak, Medine Cumhur Cure, Serap Baydur Sahin, and Suleyman Yuce Copyright © 2015 Ayse Erturk et al. All rights reserved. The Diagnosis of Invasive and Noninvasive Pulmonary Aspergillosis by Serum and Bronchoalveolar Lavage Fluid Galactomannan Assay Thu, 22 Jan 2015 11:29:36 +0000 The incidence and mortality of invasive pulmonary aspergillosis (IPA) are rising, particularly in critically ill patients and patients with severe chronic obstructive pulmonary disease (COPD). Noninvasive aspergillosis occurring in these patients requires special attention because of the possibility of developing subsequent IPA, given the poor health and worsened immune state of these patients. We compared the performance of the Platelia galactomannan (GM) enzyme immunoassay in the bronchoalveolar lavage fluid (BALF) and serum. The sensitivity, and specificity of BALF-GM were 85.4% and 62.4%, and those of serum-GM were 67.9% and 93.5% at the cutoff index of 0.5. As the cutoff index increased, the specificity of BALF-GM detection was increased with the detriment of sensitivity. The area under the ROC curves was 0.817 (95% CI: 0.718–0.916) for BALF-GM and 0.819 (95% CI: 0.712–0.926) for serum-GM. The optimal cutoff index was 1.19 for BALF-GM, and the sensitivity and specificity were 67.9% and 89.2%. The BALF-GM assay is more sensitive in detecting pulmonary aspergillosis than serum-GM assay and fungal cultures. However, BALF-GM assay has a high false-positive rate at the cutoff index of 0.5. Hence, the diagnostic cutoff index of the BALF-GM assay should be improved to avoid the overdiagnosis of pulmonary aspergillosis in clinic. Shuzhen Zhang, Sibu Wang, Zhe Wan, Ruoyu Li, and Jin Yu Copyright © 2015 Shuzhen Zhang et al. All rights reserved. CD8+ T-Cells Count in Acute Myocardial Infarction in HIV Disease in a Predominantly Male Cohort Mon, 19 Jan 2015 12:07:35 +0000 Human Immunodeficiency Virus- (HIV-) infected persons have a higher risk for acute myocardial infarction (AMI) than HIV-uninfected persons. Earlier studies suggest that HIV viral load, CD4+ T-cell count, and antiretroviral therapy are associated with cardiovascular disease (CVD) risk. Whether CD8+ T-cell count is associated with CVD risk is not clear. We investigated the association between CD8+ T-cell count and incident AMI in a cohort of 73,398 people (of which 97.3% were men) enrolled in the U.S. Veterans Aging Cohort Study-Virtual Cohort (VACS-VC). Compared to uninfected people, HIV-infected people with high baseline CD8+ T-cell counts (>1065 cells/mm3) had increased AMI risk (adjusted , 95% CI: 1.46 to 2.28). There was evidence that the effect of CD8+ T-cell tertiles on AMI risk differed by CD4+ T-cell level: compared to uninfected people, HIV-infected people with CD4+ T-cell counts ≥200 cells/mm3 had increased AMI risk with high CD8+ T-cell count, while those with CD4+ T-cell counts <200 cells/mm3 had increased AMI risk with low CD8+ T-cell count. CD8+ T-cell counts may add additional AMI risk stratification information beyond that provided by CD4+ T-cell counts alone. Oluwatosin A. Badejo, Chung-Chou Chang, Kaku A. So-Armah, Russell P. Tracy, Jason V. Baker, David Rimland, Adeel A. Butt, Adam J. Gordon, Charles R. Rinaldo Jr., Kevin Kraemer, Jeffrey H. Samet, Hilary A. Tindle, Matthew B. Goetz, Maria C. Rodriguez-Barradas, Roger Bedimo, Cynthia L. Gibert, David A. Leaf, Lewis H. Kuller, Steven G. Deeks, Amy C. Justice, and Matthew S. Freiberg Copyright © 2015 Oluwatosin A. Badejo et al. All rights reserved. Assessment of Duration of Staying Free from Acquiring Rehappening Opportunistic Infections among Pre-ART People Living with HIV/AIDS between 2008 and 2013 Sun, 18 Jan 2015 12:59:30 +0000 Introduction. In regional state of the study area, HIV (Human Immunodeficiency Virus) prevalence is 2.2% and opportunistic infections (OIs) occurred in 88.9% of pre-ART (Antiretroviral Therapy) people living with HIV/AIDS (PLWHA). Even though OIs are prevalent in the study area, duration of staying free from acquiring rehappening opportunistic infections and its determinant factors are not studied. Method. The study was conducted in randomly selected 341 adult Pre-ART PLWHA who are included in chronic HIV care. OI free duration was estimated using the actuarial life table and Kaplan Meier survival. Cox proportional-hazard model was used to calculate hazard rate. Result. OIs were rediagnosed in three quarters (75.37%) participants. In each week the probability of getting new recurrence OI was about 15.04 per 1000 person weeks. The median duration of not acquiring OI recurrence was 54 weeks. After adjustment, variables associated with recurrence were employment status, marital status, exposure for prophylaxis and adherence to it, CD4 count, and hemoglobin value. Conclusion. Giving prophylaxis and counseling to adhere it, rise in CD4 and hemoglobin level, and enhancing job opportunities should be given for PLWHA who are on chronic HIV care while continuing the care. Habtamu Mellie Bizuayehu, Direslgne Misker Abyu, and Amlaku Mulat Aweke Copyright © 2015 Habtamu Mellie Bizuayehu et al. All rights reserved. Benzodiazepines and Z-Drug Use among HIV-Infected Patients in Taiwan: A 13-Year Nationwide Cohort Study Tue, 13 Jan 2015 11:03:00 +0000 Introduction. Benzodiazepines (BZDs) and zolpidem, zopiclone, and zaleplon (Z-drugs) are commonly prescribed to HIV-infected patients. We hypothesized that frequent BZD and Z-drug use among these patients may be associated with psychiatric illnesses, particularly in long-term users. Methods. We included 1,081 patients with HIV between 1998 and 2011 from the Taiwan National Health Insurance Research Database and matched them according to age, sex, and comorbidity with uninfected controls to investigate the psychiatric diagnoses and prescriptions of BZDs and Z-drugs. Cumulative defined daily dose (cDDD) was assessed as the indicator of the duration of medication exposure. Patients exhibiting a cDDD exceeding 180 were defined as long-term users. Results. The patients with HIV had an increased risk of any use (odds ratio (OR): 8.70, 95% confidence interval (CI): 6.82–10.97) and long-term use (OR: 5.06, 95% CI: 3.63–7.04) of BZD and Z-drugs compared with those without HIV during the follow-up after demographic data and psychiatric comorbidities were adjusted. Conclusion. A large proportion of the HIV-infected patients received prescriptions of BZDs and Z-drugs. Mood disorders, insomnia, anxiety disorders, HIV infection, and substance use disorder were substantial predictors among the BZD and Z-drug users. These findings suggest that providing psychiatric services for HIV-infected patients is vital. Han-Ting Wei, Mu-Hong Chen, Wing-Wai Wong, Yuan-Hwa Chou, Ying-Jay Liou, Tung-Ping Su, Tzeng-Ji Chen, and Ya-Mei Bai Copyright © 2015 Han-Ting Wei et al. All rights reserved. Parasites: From Source to Vector and Human Sun, 21 Dec 2014 06:11:27 +0000 Veeranoot Nissapatorn, Nongyao Sawangjaroen, Rogan Lee, and Subhash Chandra Parija Copyright © 2014 Veeranoot Nissapatorn et al. All rights reserved. Mycobacterium avium subsp. hominissuis Infection in Swine Associated with Peat Used for Bedding Mon, 15 Sep 2014 06:46:57 +0000 Mycobacterium avium subsp. hominissuis is an environmental bacterium causing opportunistic infections in swine, resulting in economic losses. Additionally, the zoonotic aspect of such infections is of concern. In the southeastern region of Norway in 2009 and 2010, an increase in condemnation of pig carcasses with tuberculous lesions was seen at the meat inspection. The use of peat as bedding in the herds was suspected to be a common factor, and a project examining pigs and environmental samples from the herds was initiated. Lesions detected at meat inspection in pigs originating from 15 herds were sampled. Environmental samples including peat from six of the herds and from three peat production facilities were additionally collected. Samples were analysed by culture and isolates genotyped by MLVA analysis. Mycobacterium avium subsp. hominissuis was detected in 35 out of 46 pigs, in 16 out of 20 samples of peat, and in one sample of sawdust. MLVA analysis demonstrated identical isolates from peat and pigs within the same farms. Polyclonal infection was demonstrated by analysis of multiple isolates from the same pig. To conclude, the increase in condemnation of porcine carcasses at slaughter due to mycobacteriosis seemed to be related to untreated peat used as bedding. Tone Bjordal Johansen, Angelika Agdestein, Bjørn Lium, Anne Jørgensen, and Berit Djønne Copyright © 2014 Tone Bjordal Johansen et al. All rights reserved. Cigarette Smoke-Exposed Candida albicans Increased Chitin Production and Modulated Human Fibroblast Cell Responses Thu, 11 Sep 2014 08:27:57 +0000 The predisposition of cigarette smokers for development of respiratory and oral bacterial infections is well documented. Cigarette smoke can also contribute to yeast infection. The aim of this study was to investigate the effect of cigarette smoke condensate (CSC) on C. albicans transition, chitin content, and response to environmental stress and to examine the interaction between CSC-pretreated C. albicans and normal human gingival fibroblasts. Following exposure to CSC, C. albicans transition from blastospore to hyphal form increased. CSC-pretreated yeast cells became significantly () sensitive to oxidation but significantly () resistant to both osmotic and heat stress. CSC-pretreated C. albicans expressed high levels of chitin, with 2- to 8-fold recorded under hyphal conditions. CSC-pretreated C. albicans adhered better to the gingival fibroblasts, proliferated almost three times more and adapted into hyphae, while the gingival fibroblasts recorded a significantly () slow growth rate but a significantly higher level of IL-1β when in contact with CSC-pretreated C. albicans. CSC was thus able to modulate both C. albicans transition through the cell wall chitin content and the interaction between C. albicans and normal human gingival fibroblasts. These findings may be relevant to fungal infections in the oral cavity in smokers. Humidah Alanazi, Abdelhabib Semlali, Laura Perraud, Witold Chmielewski, Andrew Zakrzewski, and Mahmoud Rouabhia Copyright © 2014 Humidah Alanazi et al. All rights reserved. Model of Break-Bone Fever via Beta-Derivatives Thu, 11 Sep 2014 06:16:02 +0000 Using the new derivative called beta-derivative, we modelled the well-known infectious disease called break-bone fever or the dengue fever. We presented the endemic equilibrium points under certain conditions of the physical parameters included in the model. We made use of an iteration method to solve the extended model. To show the efficiency of the method used, we have presented in detail the stability and the convergence of the method for solving the system (2). We presented the uniqueness of the special solution of system (2) and finally the numerical simulations were presented for various values of beta. Abdon Atangana and Suares Clovis Oukouomi Noutchie Copyright © 2014 Abdon Atangana and Suares Clovis Oukouomi Noutchie. All rights reserved. On the Mathematical Analysis of Ebola Hemorrhagic Fever: Deathly Infection Disease in West African Countries Thu, 11 Sep 2014 06:12:34 +0000 For a given West African country, we constructed a model describing the spread of the deathly disease called Ebola hemorrhagic fever. The model was first constructed using the classical derivative and then converted to the generalized version using the beta-derivative. We studied in detail the endemic equilibrium points and provided the Eigen values associated using the Jacobian method. We furthered our investigation by solving the model numerically using an iteration method. The simulations were done in terms of time and beta. The study showed that, for small portion of infected individuals, the whole country could die out in a very short period of time in case there is not good prevention. Abdon Atangana and Emile Franc Doungmo Goufo Copyright © 2014 Abdon Atangana and Emile Franc Doungmo Goufo. All rights reserved. Highlight on Advances in Nontuberculous Mycobacterial Disease in North America Thu, 11 Sep 2014 00:00:00 +0000 Nontuberculous mycobacteria (NTM) are ubiquitous in the environment and exist as an important cause of pulmonary infections in humans. Pulmonary involvement is the most common disease manifestation of NTM and the incidence of NTM is growing in North America. Susceptibility to NTM infection is incompletely understood; therefore preventative tools are not well defined. Treatment of pulmonary nontuberculous mycobacterial (NTM) infection is difficult and entails multiple antibiotics and an extended treatment course. Also, there is a considerable variation in treatment management that should be considered before initiating treatment. We highlight the new findings in the epidemiology diagnosis and treatment of mycobacterial infections. We debate new advances regarding NTM infection in cystic fibrosis patients and solid organ transplant recipients. Finally, we introduce a new epidemiologic model for NTM disease based on virulence-exposure-host factors. Mehdi Mirsaeidi, Maham Farshidpour, Mary Beth Allen, Golnaz Ebrahimi, and Joseph O. Falkinham Copyright © 2014 Mehdi Mirsaeidi et al. All rights reserved. A Derived Network-Based Interferon-Related Signature of Human Macrophages Responding to Mycobacterium tuberculosis Sun, 31 Aug 2014 08:10:43 +0000 Network analysis of transcriptional signature typically relies on direct interaction between two highly expressed genes. However, this approach misses indirect and biological relevant interactions through a third factor (hub). Here we determine whether a hub-based network analysis can select an improved signature subset that correlates with a biological change in a stronger manner than the original signature. We have previously reported an interferon-related transcriptional signature (THP1r2Mtb-induced) from Mycobacterium tuberculosis (M. tb)-infected THP-1 human macrophage. We selected hub-connected THP1r2Mtb-induced genes into the refined network signature TMtb-iNet and grouped the excluded genes into the excluded signature TMtb-iEx. TMtb-iNet retained the enrichment of binding sites of interferon-related transcription factors and contained relatively more interferon-related interacting genes when compared to THP1r2Mtb-induced signature. TMtb-iNet correlated as strongly as THP1r2Mtb-induced signature on a public transcriptional dataset of patients with pulmonary tuberculosis (PTB). TMtb-iNet correlated more strongly in CD4+ and CD8+ T cells from PTB patients than THP1r2Mtb-induced signature and TMtb-iEx. When TMtb-iNet was applied to data during clinical therapy of tuberculosis, it resulted in the most pronounced response and the weakest correlation. Correlation on dataset from patients with AIDS or malaria was stronger for TMtb-iNet, indicating an involvement of TMtb-iNet in these chronic human infections. Collectively, the significance of this work is twofold: (1) we disseminate a hub-based approach in generating a biologically meaningful and clinically useful signature; (2) using this approach we introduce a new network-based signature and demonstrate its promising applications in understanding host responses to infections. Kang Wu, Hai Fang, Liang-Dong Lyu, Douglas B. Lowrie, Ka-Wing Wong, and Xiao-Yong Fan Copyright © 2014 Kang Wu et al. All rights reserved. Proteinases in Excretory-Secretory Products of Toxocara canis Second-Stage Larvae: Zymography and Modeling Insights Thu, 14 Aug 2014 06:49:17 +0000 Components released in excretory-secretory products of Toxocara canis larvae (TES) include phosphatidylethanolamine-binding proteins (TES26), mucins (TES120, MUC2-5), and C-type lectins (TES32, TES70) and their biochemical, immunological, and diagnostic properties have been extensively studied albeit proteinase activities towards physiological substrates are almost unknown. Proteolytic activities in TES samples were first analyzed by gel electrophoresis with gelatin as substrate. Major activities of ~400, 120, and 32 kDa in TES were relatively similar over a broad pH range (5.5–9.0) and all these were of the serine-type as leupeptin abolished gelatinolysis. Further, the ~400 kDa component degraded all physiological substrates tested (laminin, fibronectin, albumin, and goat IgG) and the 120 kDa component degraded albumin and goat IgG while proteinases of lower MW (45, 32, and 26 kDa) only degraded laminin and fibronectin, preferentially at alkaline pH (9.0). By protein modeling approaches using the known sequences of TES components, only TES26 and MUC4 displayed folding patterns significantly related to reference serine proteinases. These data suggest that most of serine proteinase activities secreted in vitro by infective larvae of T. canis have intriguing nature but otherwise help the parasite to affect multiple components of somatic organs and bodily fluids within the infected host. Gonzalo Ernesto González-Páez, Fernando Alba-Hurtado, Carlos Gerardo García-Tovar, and Raúl Argüello-García Copyright © 2014 Gonzalo Ernesto González-Páez et al. All rights reserved. Evaluation of Novel Tools to Facilitate the Detection and Characterization of Leprosy Patients in China Tue, 12 Aug 2014 08:57:53 +0000 Leprosy is the disabling outcome of chronic infection with Mycobacterium leprae. The disease often evades early detection, particularly now that fewer clinicians are able to confidently diagnose the disease following the integration of leprosy control measures within general health services in many countries. Although leprosy is officially eliminated in China, endemic regions remain in some difficult-to-reach, underdeveloped areas in Southwest China. In order to better understand the extent of M. leprae infection and identify new leprosy cases in a timely manner, simple tools that can detect infection and the early disease are required. In this report we evaluated the performance of antigen-specific ELISA, the NDO-LID rapid diagnostic test, and antigen-specific whole blood assays (WBA) as potential diagnostic tools. Our data support the use of antibody detection tests and WBA to facilitate the diagnosis of multibacillary and paucibacillary leprosy, respectively. These tools could be invaluable for increased, but simplified, monitoring of individuals in order to provide referrals for clinical exam and early leprosy diagnosis. Yan Wen, Yuan Gang You, Lian-Chao Yuan, You Hua Yuan, Ying Zhang, Malcolm S. Duthie, and Huan-Ying Li Copyright © 2014 Yan Wen et al. All rights reserved. Prevalence of Ischemic Heart Disease and Management of Coronary Risk in Daily Clinical Practice: Results from a Mediterranean Cohort of HIV-Infected Patients Thu, 07 Aug 2014 10:55:02 +0000 Background. There are conflicting data on the prevalence of coronary events and the quality of the management of modifiable cardiovascular risk factors (CVRF) in HIV-infected patients. Methods. We performed a retrospective descriptive study to determine the prevalence of coronary events and to evaluate the management of CVRF in a Mediterranean cohort of 3760 HIV-1-infected patients from April 1983 through June 2011. Results. We identified 81 patients with a history of a coronary event (prevalence 2.15%); 83% of them suffered an acute myocardial infarction. At the time of the coronary event, CVRF were highly prevalent (60.5% hypertension, 48% dyslipidemia, and 16% diabetes mellitus). Other CVRF, such as smoking, hypertension, lack of exercise, and body mass index, were not routinely assessed. After the coronary event, a significant decrease in total cholesterol and LDL-cholesterol was observed. However, the percentage of patients who maintained LDL-cholesterol > 100 mg/dL remained stable (from 46% to 41%, ). Patients using protease inhibitors associated with a favorable lipid profile increased over time . Conclusions. The prevalence of coronary events in our cohort is low. CVRF prevalence is high and their management is far from optimal. More aggressive interventions should be implemented to diminish cardiovascular risk in HIV-infected patients. Patricia Echeverría, Pere Domingo, Josep-María Llibre, Mar Gutierrez, Gracia Mateo, Jordi Puig, Anna Bonjoch, Nuria Pérez-Alvarez, Guillem Sirera, Bonaventura Clotet, and Eugenia Negredo Copyright © 2014 Patricia Echeverría et al. All rights reserved. Description of a Novel Adhesin of Mycobacterium avium Subsp. paratuberculosis Tue, 22 Jul 2014 08:13:46 +0000 The binding and ingestion of Mycobacterium avium subsp. paratuberculosis (MAP) by host cells are fibronectin (FN) dependent. In several species of mycobacteria, a specific family of proteins allows the attachment and internalization of these bacteria by epithelial cells through interaction with FN. Thus, the identification of adhesion molecules is essential to understand the pathogenesis of MAP. The aim of this study was to identify and characterize FN binding cell wall proteins of MAP. We searched for conserved adhesins within a large panel of surface immunogenic proteins of MAP and investigated a possible interaction with FN. For this purpose, a cell wall protein fraction was obtained and resolved by 2D electrophoresis. The immunoreactive spots were identified by MALDI-TOF MS and a homology search was performed. We selected elongation factor Tu (EF-Tu) as candidate for further studies. We demonstrated the FN-binding capability of EF-Tu using a ligand blot assay and also confirmed the interaction with FN in a dose-dependent manner by ELISA. The dissociation constant of EF-Tu was determined by surface plasmon resonance and displayed values within the μM range. These data support the hypothesis that this protein could be involved in the interaction of MAP with epithelial cells through FN binding. Mariana Noelia Viale, Gabriela Echeverria-Valencia, Pablo Romasanta, María Laura Mon, Marisa Fernandez, Emilio Malchiodi, María Isabel Romano, Andrea Karina Gioffré, and María de la Paz Santangelo Copyright © 2014 Mariana Noelia Viale et al. All rights reserved. Event-Specific Risk Factors Predicting Episodes of Unprotected Anal Intercourse with Male Nonregular Partners among Men Who Have Sex with Men Using Case-Crossover Study Design Sun, 20 Jul 2014 12:06:45 +0000 This study investigated event-specific factors that determine episodes of unprotected and protected anal intercourse (UAI and PAI) among 215 men who have sex with men (MSM), who used condoms inconsistently with nonregular partners (NRP) in the last six months, in Hong Kong. A case-crossover study design was used. Lower likelihood of episodes involving UAI with NRP was associated with (1) five partner attributes (NRP were <35 years old, at least three previous anal sex experiences with the NRP, perception that participant and the NRP had asymmetrical sexual experience, perception that the NRP was feminine, and liking toward the NRP; OR = 0.16–0.52), (2) six situational variables (the participant having had UAI with another man in the last week, having discussed condom use, perception that the NRP liked to use condom, partner’s suggestion to have PAI, participant’s suggestion to have PAI, and participant’s plan to use condoms; OR = 0.11–0.39), and (3) four environmental/setting variables (condoms already placed at the venue, display of condom use promotion materials, participant’s possession of a condom, and the NRP possessed a condom; OR = 0.27–0.45). HIV prevention targeting MSM should focus on event-specific protective factors, which may be different from those obtained from studies distinguishing condom users versus nonusers. Jinghua Li, Joseph T. F. Lau, Jing Gu, Chun Hao, and Coco H. Y. Lai Copyright © 2014 Jinghua Li et al. All rights reserved. Epidemiology, Phylogeny, and Evolution of Emerging Enteric Picobirnaviruses of Animal Origin and Their Relationship to Human Strains Thu, 17 Jul 2014 12:02:44 +0000 Picobirnavirus (PBV) which has been included in the list of viruses causing enteric infection in animals is highly versatile because of its broad host range and genetic diversity. PBVs are among the most recent and emerging small, nonenveloped viruses with a bisegmented double-stranded RNA genome, classified under a new family “Picobirnaviridae.” PBVs have also been detected from respiratory tract of pigs, but needs further close investigation for their inhabitant behavior. Though, accretion of genomic data of PBVs from different mammalian species resolved some of the ambiguity, quite a few questions and hypotheses regarding pathogenesis, persistence location, and evolution of PBVs remain unreciprocated. Evolutionary analysis reveals association of PBVs with partitiviruses especially fungi partitiviruses. Although, PBVs may have an ambiguous clinical implication, they do pose a potential public health concern in humans and control of PBVs mainly relies on nonvaccinal approach. Based upon the published data, from 1988 to date, generated from animal PBVs across the globe, this review provides information and discussion with respect to genetic analysis as well as evolution of PBVs of animal origin in relation to human strains. Yashpal S. Malik, Naveen Kumar, Kuldeep Sharma, Kuldeep Dhama, Muhammad Zubair Shabbir, Balasubramanian Ganesh, Nobumichi Kobayashi, and Krisztian Banyai Copyright © 2014 Yashpal S. Malik et al. All rights reserved. Computational Analysis of the Model Describing HIV Infection of CD4+T Cells Thu, 17 Jul 2014 00:00:00 +0000 An analysis of the model underpinning the description of the spread of HIV infection of CD4+T cells is examined in detail in this work. Investigations of the disease free and endemic equilibrium are done using the method of Jacobian matrix. An iteration technique, namely, the homotopy decomposition method (HDM), is implemented to give an approximate solution of nonlinear ordinary differential equation systems. The technique is described and illustrated with numerical examples. The approximated solution obtained via HDM is compared with those obtained via other methods to prove the trustworthiness of HDM. Moreover, the lessening and simplicity in calculations furnish HDM with a broader applicability. Abdon Atangana and Emile Franc Doungmo Goufo Copyright © 2014 Abdon Atangana and Emile Franc Doungmo Goufo. All rights reserved. Harnessing the Helminth Secretome for Therapeutic Immunomodulators Tue, 15 Jul 2014 09:19:29 +0000 Helminths are the largest and most complex pathogens to invade and live within the human body. Since they are not able to outpace the immune system by rapid antigen variation or faster cell division or retreat into protective niches not accessible to immune effector mechanisms, their long-term survival depends on influencing and regulating the immune responses away from the mode of action most damaging to them. Immunologists have focused on the excretory and secretory products that are released by the helminths, since they can change the host environment by modulating the immune system. Here we give a brief overview of the helminth-associated immune response and the currently available helminth secretome data. We introduce some major secretome-derived immunomodulatory molecules and describe their potential mode of action. Finally, the applicability of helminth-derived therapeutic proteins in the treatment of allergic and autoimmune inflammatory disease is discussed. Dana Ditgen, Emmanuela M. Anandarajah, Kamila A. Meissner, Norbert Brattig, Carsten Wrenger, and Eva Liebau Copyright © 2014 Dana Ditgen et al. All rights reserved. First Record of Isolation and Characterization of Methicillin Resistant Staphylococcus lugdunensis from Clinical Samples in Iraq Sun, 13 Jul 2014 11:08:32 +0000 This study was conducted to determine the frequency of Staphylococcus lugdunensis in different clinical samples. Out of 690 clinical samples, a total of 178 coagulase negative staphylococci (CoNS) isolates were recovered. CoNS were identified as 10 different species; 22 isolates belonged to Staphylococcus lugdunensis. Two specific genes for S. lugdunensis were used ( tanA gene and fbl gene) to confirm identification. Both of these specific genes were detected in 15 (68.1%) of 22 isolates that were identified phenotypically. The results of oxacillin MIC showed that 7 of the 15 (46.6%) S. lugdunensis isolates were oxacillin resistant. The antibiotic susceptibility testing against 16 antibiotics showed that resistance rates were variable towards these antibiotics. Eight of fifteen S. lugdunensis isolates (53.3%) were β-lactamase producer. Results of molecular detection of mecA gene found that mecA gene was detected in 6 (40%) of 15 S. lugdunensis. All of these 6 isolates (S1, S2, S3, S4, S5, and S6) were resistant to oxacillin. One isolate (S7) was resistant to oxacillin but mecA was not detected in this isolate. This study is a first record of isolation and characterization of methicillin resistant S. lugdunensis (MRSL) from clinical samples in Iraq. Alaa H. Al-Charrakh and Mohammed H. Obayes Copyright © 2014 Alaa H. Al-Charrakh and Mohammed H. Obayes. All rights reserved. Mycobacterium tuberculosis P-Type ATPases: Possible Targets for Drug or Vaccine Development Thu, 10 Jul 2014 11:38:46 +0000 Tuberculosis (TB) has been the biggest killer in the human history; currently, Mycobacterium tuberculosis (Mtb) kills nearly 2 million people each year worldwide. The high prevalence of TB obligates the identification of new therapeutic targets and the development of anti-TB vaccines that can control multidrug resistance and latent TB infections. Membrane proteins have recently been suggested as key targets for bacterial viability. Current studies have shown that mycobacteria P-type ATPases may play critical roles in ion homeostasis and in the response of mycobacteria to toxic substances in the intraphagosomal environment. In this review, we bring together the genomic, transcriptomic, and structural aspects of the P-type ATPases that are relevant during active and latent Mtb infections, which can be useful in determining the potential of these ATPases as drug targets and in uncovering their possible roles in the development of new anti-TB attenuated vaccines. Lorena Novoa-Aponte and Carlos Yesid Soto Ospina Copyright © 2014 Lorena Novoa-Aponte and Carlos Yesid Soto Ospina. All rights reserved. Modelling the Aggregation Process of Cellular Slime Mold by the Chemical Attraction Tue, 08 Jul 2014 08:54:41 +0000 We put into exercise a comparatively innovative analytical modus operandi, the homotopy decomposition method (HDM), for solving a system of nonlinear partial differential equations arising in an attractor one-dimensional Keller-Segel dynamics system. Numerical solutions are given and some properties show evidence of biologically practical reliance on the parameter values. The reliability of HDM and the reduction in computations give HDM a wider applicability. Abdon Atangana and P. D. Vermeulen Copyright © 2014 Abdon Atangana and P. D. Vermeulen. All rights reserved. Evaluation of Cocktails with Recombinant Proteins of Mycobacterium bovis for a Specific Diagnosis of Bovine Tuberculosis Tue, 08 Jul 2014 08:16:15 +0000 The Delayed type hypersensitivity skin test (DTH) and interferon-gamma assay are used for the diagnosis of bovine tuberculosis (TBB). The specificity of these diagnoses, however, is compromised because both are based on the response against purified protein derivative of Mycobacterium bovis (PPD-B). In this study, we assessed the potential of two cocktails containing M. bovis recombinant proteins: cocktail 1 (C1): ESAT-6, CFP-10 and MPB83 and cocktail 2 (C2): ESAT-6, CFP-10, MPB83, HspX, TB10.3, and MPB70. C1, C2, and PPD-B showed similar response by DTH in M. bovis-sensitized guinea pigs. Importantly, C1 induced a lower response than PPD-B in M. avium-sensitized guinea pigs. In cattle, C1 displayed better performance than PPD-B and C2; indeed, C1 showed the least detection of animals either vaccinated or Map-infected. To optimize the composition of the cocktails, we obtained protein fractions from PPD-B and tested their immunogenicity in experimentally M. bovis-infected cattle. In one highly reactive fraction, seven proteins were identified. The inclusion of FixB in C1 enhanced the recognition of M. bovis-infected cattle without compromising specificity. Our data provide a promising basis for the future development of a cocktail for TBB detection without interference by the presence of sensitized or infected animals with other mycobacteria. María Laura Mon, Roberto Damián Moyano, Mariana Noelia Viale, María Alejandra Colombatti Olivieri, Ignacio José Gamietea, Valeria Noely Montenegro, Bernardo Alonso, María de la Paz Santangelo, Mahavir Singh, Rosario Duran, and María Isabel Romano Copyright © 2014 María Laura Mon et al. All rights reserved. Phylogenetic and In Silico Functional Analyses of Thermostable-Direct Hemolysin and tdh-Related Encoding Genes in Vibrio parahaemolyticus and Other Gram-Negative Bacteria Tue, 08 Jul 2014 06:54:25 +0000 Emergence and spread of pandemic strains of Vibrio parahaemolyticus have drawn attention to make detailed study on their genomes. The pathogenicity of V. parahaemolyticus has been associated with thermostable-direct hemolysin (TDH) and/or TDH-related hemolysin (TRH). The present study evaluated characteristics of tdh and trh genes, considering the phylogenetic and in silico functional features of V. parahaemolyticus and other bacteria. Fifty-two tdh and trh genes submitted to the GenBank were analyzed for sequence similarity. The promoter sequences of these genes were also analyzed from transcription start point to −35 regions and correlated with amino acid substitution within the coding regions. The phylogenetic analysis revealed that tdh and trh are highly distinct and also differ within the V. parahaemolyticus strains that were isolated from different geographical regions. Promoter sequence analysis revealed nucleotide substitutions and deletions at −18 and −19 positions among the pandemic, prepandemic, and nonpandemic tdh sequences. Many amino acid substitutions were also found within the signal peptide and also in the matured protein region of several TDH proteins as compared to TDH-S protein of pandemic V. parahaemolyticus. Experimental evidences are needed to recognize the importance of substitutions and deletions in the tdh and trh genes. Sushanta K. Bhowmik, Gururaja P. Pazhani, and Thandavarayan Ramamurthy Copyright © 2014 Sushanta K. Bhowmik et al. All rights reserved. Differential Expression of Immunogenic Proteins on Virulent Mycobacterium tuberculosis Clinical Isolates Mon, 07 Jul 2014 00:00:00 +0000 Molecular epidemiology has revealed that Mycobacterium tuberculosis (Mtb), formerly regarded as highly conserved species, displays a considerable degree of genetic variability that can influence the outcome of the disease as well as the innate and adaptive immune response. Recent studies have demonstrated that Mtb families found worldwide today differ in pathology, transmissibility, virulence, and development of immune response. By proteomic approaches seven proteins that were differentially expressed between a local clinical isolate from Latin-American-Mediterranean (LAM) and from Haarlem (H) lineages were identified. In order to analyze the immunogenic ability, recombinant Rv2241, Rv0009, Rv0407, and Rv2624c proteins were produced for testing specific antibody responses. We found that these proteins induced humoral immune responses in patients with drug-sensitive and drug-resistant tuberculosis with substantial cross-reactivity among the four proteins. Moreover, such reactivity was also correlated with anti-Mtb-cell surface IgM, but not with anti-ManLAM, anti-PPD, or anti-Mtb-surface IgG antibodies. Therefore, the present results describe new Mtb antigens with potential application as biomarkers of TB. Pablo Schierloh, Laura Klepp, Camila Vazquez, Roxana Valeria Rocha, Federico Carlos Blanco, Luciana Balboa, Beatriz López, Viviana Ritacco, Fabiana Bigi, and María del Carmen Sasiain Copyright © 2014 Pablo Schierloh et al. All rights reserved. Induction of Boosted Immune Response in Mice by Leptospiral Surface Proteins Expressed in Fusion with DnaK Sun, 06 Jul 2014 06:50:41 +0000 Leptospirosis is an important global disease of human and veterinary concern. Caused by pathogenic Leptospira, the illness was recently classified as an emerging infectious disease. Currently available veterinarian vaccines do not induce long-term protection against infection and do not provide cross-protective immunity. Several studies have suggested the use of DnaK as an antigen in vaccine formulation, due to an exceptional degree of immunogenicity. We focused on four surface proteins: rLIC10368 (Lsa21), rLIC10494, rLIC12690 (Lp95), and rLIC12730, previously shown to be involved in host-pathogen interactions. Our goal was to evaluate the immunogenicity of the proteins genetically fused with DnaK in animal model. The chosen genes were amplified by PCR methodology and cloned into pAE, an E. coli vector. The recombinant proteins were expressed alone or in fusion with DnaK at the N-terminus. Our results demonstrate that leptospiral proteins fused with DnaK have elicited an enhanced immune response in mice when compared to the effect promoted by the individual proteins. The boosted immune effect was demonstrated by the production of total IgG, lymphocyte proliferation, and significant amounts of IL-10 in supernatant of splenocyte cell cultures. We believe that this approach could be employed in vaccines to enhance presentation of antigens of Leptospira to professional immune cells. Marina V. Atzingen, Dunia Rodriguez, Gabriela Hase Siqueira, Luciana C. C. Leite, and Ana L. T. O. Nascimento Copyright © 2014 Marina V. Atzingen et al. All rights reserved. Low Prevalence of Hepatitis B and C Virus Markers among Children and Adolescents Tue, 01 Jul 2014 07:57:13 +0000 This study aimed to determine the prevalence of HBV and HCV among children and adolescents attending schools and daycare centres in Rio de Janeiro State, located in southern Brazil. Serum samples from 1,217 individuals aged 0 to 18 years were collected from 1999 to 2012 and tested for HBsAg, total anti-HBc, anti-HBs, and anti-HCV by ELISA. Reactive HBsAg and anti-HBc samples were tested for HBV DNA. Reactive anti-HCV samples were tested for HCV RNA and genotyped by RFLP. HBsAg was detected in 1.8% of individuals, and total anti-HBc was detected among 3.6% of individuals. Anti-HBs reactivity was found among 25.3% (322/1,217) of the individuals and increased from 6.28% in the years 1999-2000 to 76.2% in the years 2001–2012 . HBV DNA was detected in 18 of 51 individuals who presented with HBsAg or isolated anti-HBc, and nine were considered occult hepatitis B cases. Three individuals were anti-HCV- and HCV RNA-positive: two of them were infected with genotype 1, and the other was infected with genotype 3. Low levels of HBV and HCV markers were observed in children and adolescents. HBV immunity increased during the period of study, indicating that childhood universal HBV vaccination has been effective for controlling HBV infection in Brazil. Livia Melo Villar, Luciane Almeida Amado, Adilson José de Almeida, Vanessa Salete de Paula, Lia Laura Lewis-Ximenez, and Elisabeth Lampe Copyright © 2014 Livia Melo Villar et al. All rights reserved. Analysis of Antigens of Mycobacterium leprae by Interaction to Sera IgG, IgM, and IgA Response to Improve Diagnosis of Leprosy Sun, 29 Jun 2014 11:25:57 +0000 Till 2010, several countries have declared less than one leprosy patient among population of 10,000 and themselves feeling as eliminated from leprosy cases. However, new leprosy cases are still appearing from all these countries. In this situation one has to be confident to diagnose leprosy. This review paper highlighted already explored antigens for diagnosis purposes and finally suggested better combinations of protein antigens of M. leprae versus immunoglobulin as detector antibody to be useful for leprosy diagnosis. Avnish Kumar, Om Parkash, and Bhawneshwar K. Girdhar Copyright © 2014 Avnish Kumar et al. All rights reserved. Immunopathological Aspects of Experimental Trypanosoma cruzi Reinfections Tue, 24 Jun 2014 10:27:26 +0000 Chagas disease is caused by Trypanosoma cruzi infection. Besides the host-related factors, such as immune response and genetic background, the parasite, strain, and occurrences of reinfection episodes, may influence disease outcome. Our results demonstrate that both the primary infection and the reinfection with the Colombiana strain are connected with lower survival rate of the mice. After reinfection, parasitaemia is approximately ten times lower than in primary infected animals. Only Colombiana, Colombiana/Colombiana, and Y/Colombiana groups presented amastigote nests in cardiac tissue. Moreover, the mice infected and/or reinfected with the Colombiana strain had more T. cruzi nests, more intense inflammatory infiltrate, and higher in situ expression of TNF-α and IFN-γ than Y strain. Antigen-stimulated spleen cells from infected and/or reinfected animals produced higher levels of TNF-α, IFN-γ, and IL-10. Our results reinforce the idea that Chagas disease outcome is influenced by the strain of the infective parasite, being differentially modulated during reinfection episodes. It highlights the need of control strategies involving parasite strain characterization in endemic areas for Chagas disease. Juliana Reis Machado, Marcos Vinícius Silva, Diego Costa Borges, Crislaine Aparecida da Silva, Luis Eduardo Ramirez, Marlene Antônia dos Reis, Lúcio Roberto Castellano, Virmondes Rodrigues, and Denise Bertulucci Rocha Rodrigues Copyright © 2014 Juliana Reis Machado et al. All rights reserved. Helicobacter pylori Is Not Eradicated after Triple Therapy: A Nested PCR Based Study Mon, 23 Jun 2014 06:51:59 +0000 Detection of Helicobacter pylori after triple therapy is usually carried out by either rapid urease test (RUT), urea breath test (UBT), histology, bacterial isolation, and single round PCR or serological tests. In this study, antral biopsy specimens from 25 patients were tested for H. pylori by RUT, culture, histology, and nested PCR in their antral biopsy specimens before and after treatment. Three genes, namely, heat shock protein (hsp60), phosphoglucosamine mutase (ureC), and flagellar export ATP synthase (fliI) of H. pylori were targeted. Of the 25 antral biopsy specimens, the RUT, culture, histology, and nested PCR positivity dropped from 81.8% to 12%, 31% to 0%, 100 to 84%, and 100% to 92%, respectively, before and after therapy. Further, hsp60 specific amplicons from 23 out of 25 patients gave identical restriction pattern, while 6 fliI and 1 ureC specific amplicon produced different restriction pattern. Furthermore, variations in fliI gene sequences in H. pylori after treatment were also confirmed by sequencing and compared in silico. Nested PCR based detection of H. pylori is more sensitive method to detect H. pylori after therapy than culture, RUT, and histology. Further, this study suggests that H. pylori is not eradicated completely after triple therapy. Saurabh Kumar Patel, Girish Narayan Mishra, Chandra Bhan Pratap, Ashok Kumar Jain, and Gopal Nath Copyright © 2014 Saurabh Kumar Patel et al. All rights reserved. Regional Warming and Emerging Vector-Borne Zoonotic Dirofilariosis in the Russian Federation, Ukraine, and Other Post-Soviet States from 1981 to 2011 and Projection by 2030 Thu, 19 Jun 2014 09:15:01 +0000 We analyze through a climatic model the influence of regional warming on the geographical spreading and potential risk of infection of human dirofilariosis in Russia, Ukraine, and other post-Soviet states from 1981 to 2011 and estimate the situation by 2030. The model correctly predicts the spatiotemporal location of 97.10% of 2154 clinical cases reported in the area during the studied period, identified by a retrospective review of the literature. There exists also a significant correlation between annual predicted Dirofilaria generations and calculated morbidity. The model states the progressive increase of 14.8% in the potential transmission area, up to latitude 64°N, and 14.7% in population exposure. By 2030 an increase of 18.5% in transmission area and 10.8% in population exposure is expected. These findings strongly suggest the influence of global warming in both geographical spreading and increase in the number of Dirofilaria generations. The results should alert about the epidemiological behavior of dirofilariosis and other mosquito-borne diseases in these and other countries with similar climatic characteristics. Vladimir Kartashev, Alexandr Afonin, Javier González-Miguel, Rosa Sepúlveda, Luis Simón, Rodrigo Morchón, and Fernando Simón Copyright © 2014 Vladimir Kartashev et al. All rights reserved. Seroprevalence of Antibodies against Pkn1, a Novel Potential Immunogen, in Chlamydia trachomatis-Infected Macaca nemestrina and Human Patients Wed, 18 Jun 2014 08:26:51 +0000 Chlamydia trachomatis (CT) is an important cause of sexually transmitted genital tract infections (STIs) and trachoma. Despite major research into chlamydial pathogenesis and host immune responses, immunoprotection has been hampered by the incomplete understanding of protective immunity in the genital tract. Characterized vaccine candidates have shown variable efficacy ranging from no protection to partial protection in vivo. It is therefore a research priority to identify novel chlamydial antigens that may elicit protective immune responses against CT infection. In the present study we assessed the seroprevalence of antibodies against protein kinase1 (Pkn1), DNA ligaseA (LigA), and major outer membrane protein A (OmpA) following natural CT infection in humans and in experimentally induced CT infection in Macaca nemestrina. Antigenic stretches of Pkn1, LigA, and OmpA were identified using bioinformatic tools. Pkn1, LigA, and OmpA genes were cloned in bacterial expression vector and purified by affinity chromatography. Our results demonstrate significantly high seroprevalence of antibodies against purified Pkn1 and OmpA in sera obtained from the macaque animal model and human patients infected with CT. In contrast no significant seroreactivity was observed for LigA. The seroprevalence of antibodies against Pkn1 suggest that nonsurface chlamydial proteins could also be important for developing vaccines for C. trachomatis. Achchhe L. Patel, Prashant K. Mishra, Divya Sachdev, Uma Chaudhary, Dorothy L. Patton, and Daman Saluja Copyright © 2014 Achchhe L. Patel et al. All rights reserved. Liposomal Formulation of Turmerone-Rich Hexane Fractions from Curcuma longa Enhances Their Antileishmanial Activity Wed, 18 Jun 2014 00:00:00 +0000 Promastigote forms of Leishmania amazonensis were treated with different concentrations of two fractions of Curcuma longa cortex rich in turmerones and their respective liposomal formulations in order to evaluate growth inhibition and the minimal inhibitory concentration (MIC). In addition, cellular alterations of treated promastigotes were investigated under transmission and scanning electron microscopies. LipoRHIC and LipoRHIWC presented lower MIC, 5.5 and 12.5 μg/mL, when compared to nonencapsulated fractions (125 and 250 μg/mL), respectively, and to ar-turmerone (50 μg/mL). Parasite growth inhibition was demonstrated to be dose-dependent. Important morphological changes as rounded body and presence of several roles on plasmatic membrane could be seen on L. amazonensis promastigotes after treatment with subinhibitory concentration (2.75 μg/mL) of the most active LipoRHIC. In that sense, the hexane fraction from the turmeric cortex of Curcuma longa incorporated in liposomal formulation (LipoRHIC) could represent good strategy for the development of new antileishmanial agent. Ana Claudia F. Amaral, Luciana A. Gomes, Jefferson Rocha de A. Silva, José Luiz P. Ferreira, Aline de S. Ramos, Maria do Socorro S. Rosa, Alane B. Vermelho, and Igor A. Rodrigues Copyright © 2014 Ana Claudia F. Amaral et al. All rights reserved. High Prevalence of Panton-Valentine Leukocidin (PVL) Genes in Nosocomial-Acquired Staphylococcus aureus Isolated from Tertiary Care Hospitals in Nepal Wed, 18 Jun 2014 00:00:00 +0000 Methicillin-resistant Staphylococcus aureus (MRSA) carrying the important virulence determinant, Panton-Valentine leukocidin (PVL), is an emerging infectious pathogen associated with skin and soft tissue infections as well as life-threatening invasive diseases. In carrying out the first PVL prevalence study in Nepal, we screened 73 nosocomial isolates of S. aureus from 2 tertiary care Nepali hospitals and obtained an overall PVL-positivity rate of 35.6% among the hospital isolates: 26.1% of MRSA and 51.9% of methicillin sensitive S. aureus (MSSA) isolates were found to be positive for the PVL genes. PVL prevalence was not associated with a specific (i) infection site, (ii) age group, or (iii) hospital of origin. It was found to be positively associated with heterogeneous MRSA (73.3%) compared to homogeneous MRSA (3.2%) and MSSA (51.9%); negatively associated with multiresistant MRSA (22%) compared to nonmultiresistant MRSA (60%) and MSSA (51.9%); and positively associated with macrolide-streptogramin B resistance (93.8%) compared to macrolide-lincosamide-streptogramin B resistance (0%) and no-resistance (45.8%) types. Macrolide-streptogramin B resistance was confirmed by the presence of msr(A) gene. Restriction pattern analyses provided evidence to support the circulation of a limited number of clones of PVL-positive MRSA, arguing for the adaptability of these isolates to a hospital setting. Bidya Shrestha, Winny Singh, V. Samuel Raj, Bharat Mani Pokhrel, and Tribhuban Mohan Mohapatra Copyright © 2014 Bidya Shrestha et al. All rights reserved. Performance and Logistical Challenges of Alternative HIV-1 Virological Monitoring Options in a Clinical Setting of Harare, Zimbabwe Sun, 15 Jun 2014 05:25:06 +0000 We evaluated a low-cost virological failure assay (VFA) on plasma and dried blood spot (DBS) specimens from HIV-1 infected patients attending an HIV clinic in Harare. The results were compared to the performance of the ultrasensitive heat-denatured p24 assay (p24). The COBAS AmpliPrep/COBAS TaqMan HIV-1 test, version 2.0, served as the gold standard. Using a cutoff of 5,000 copies/mL, the plasma VFA had a sensitivity of 94.5% and specificity of 92.7% and was largely superior to the VFA on DBS (sensitivity = 61.9%; specificity = 99.0%) or to the p24 (sensitivity = 54.3%; specificity = 82.3%) when tested on 302 HIV treated and untreated patients. However, among the 202 long-term ART-exposed patients, the sensitivity of the VFA decreased to 72.7% and to 35.7% using a threshold of 5,000 and 1,000 RNA copies/mL, respectively. We show that the VFA (either on plasma or on DBS) and the p24 are not reliable to monitor long-term treated, HIV-1 infected patients. Moreover, achieving acceptable assay sensitivity using DBS proved technically difficult in a less-experienced laboratory. Importantly, the high level of virological suppression (93%) indicated that quality care focused on treatment adherence limits virological failure even when PCR-based viral load monitoring is not available. Pascale Ondoa, Tinei Shamu, Michelle Bronze, Maureen Wellington, Tamara Sonia Boender, Corry Manting, Kim Steegen, Rudi Luethy, and Tobias Rinke de Wit Copyright © 2014 Pascale Ondoa et al. All rights reserved. Directly-Observed Intermittent Therapy versus Unsupervised Daily Regimen during the Intensive Phase of Antituberculosis Therapy in HIV Infected Patients Wed, 11 Jun 2014 14:12:22 +0000 The World Health Organization strongly recommends using daily antituberculosis therapy (ATT) during the intensive phase for HIV infected patients. India has the highest burden of tuberculosis in the world, but HIV infected patients are still receiving intermittent ATT. In this study we compared the mortality in patients who received directly-observed intermittent ATT versus self-administered daily ATT with fixed dose combinations during the intensive phase in a context of freely available antiretroviral therapy. The study included 1460 patients, 343 in the intermittent ATT group and 1117 in the daily ATT group. Baseline covariates of the two groups were balanced using inverse probability of treatment weighting based on propensity score methods. In a sensitivity analysis, continuous variables (albumin, CD4 count, and age) were modelled using restricted cubic smoothing splines. Compared with patients who received daily ATT, patients who received intermittent ATT had a 40% higher risk of mortality (1.4 hazard ratio; 95% confidence interval, 1.14–1.7). We estimated that the use of daily ATT could achieve a 10% absolute reduction in mortality at 12 months. Self-administered daily ATT was not associated with an increased risk of default from treatment. These results support the immediate implementation of daily ATT for HIV infected patients during the intensive phase in India. Gerardo Alvarez-Uria, Manoranjan Midde, Raghavakalyan Pakam, and Praveen Kumar Naik Copyright © 2014 Gerardo Alvarez-Uria et al. All rights reserved. Exposure of a 23F Serotype Strain of Streptococcus pneumoniae to Cigarette Smoke Condensate Is Associated with Selective Upregulation of Genes Encoding the Two-Component Regulatory System 11 (TCS11) Wed, 11 Jun 2014 12:55:49 +0000 Alterations in whole genome expression profiles following exposure of the pneumococcus (strain 172, serotype 23F) to cigarette smoke condensate (160 μg/mL) for 15 and 60 min have been determined using the TIGR4 DNA microarray chip. Exposure to CSC resulted in the significant (–0.0006) upregulation of the genes encoding the two-component regulatory system 11 (TCS11), consisting of the sensor kinase, hk11, and its cognate response regulator, rr11, in the setting of increased biofilm formation. These effects of cigarette smoke on the pneumococcus may contribute to colonization of the airways by this microbial pathogen. Riana Cockeran, Jenny A. Herbert, Timothy J. Mitchell, Thérèse Dix-Peek, Caroline Dickens, Ronald Anderson, and Charles Feldman Copyright © 2014 Riana Cockeran et al. All rights reserved. Characterization of Shiga Toxin-Producing Escherichia coli Isolated from Ground Beef Collected in Different Socioeconomic Strata Markets in Buenos Aires, Argentina Mon, 09 Jun 2014 09:01:17 +0000 Consumption of raw/undercooked ground beef is the most common route of transmission of Shiga toxin-producing E. coli (STEC). The aim of the study was to determine the STEC contamination level of the ground beef samples collected in 36 markets of different socioeconomic strata in Buenos Aires, Argentina, and the characterization of the isolated strains. Ninety-one out of 252 (36.1%) samples were stx+. Fifty-seven STEC strains were recovered. Eleven STEC strains belonged to O157 serogroup, and 46 to non-O157 serogroups. Virulence markers of the 57 STEC were stx1, 5.3% (3/57); stx2, 86.0% (49/57); stx1/stx2, 8.8% (5/57); ehxA, 61.4% (35/57); eae, 26.3% (15/57); saa, 24.6% (14/57). Shiga toxin subtypes were stx2, 31.5% (17/54); stx2c-vhb, 24.1% (13/54); stx2c-vha, 20.4% (11/54); stx2/stx2c-vha, 14.8% (8/54); stx2/stx2c-vhb, 5.6% (3/54); stx2c-vha/vhb, 3.7% (2/54). Serotypes O178:H19 and O157:H7 were prevalent. Contamination rate of STEC in all strata was high, and the highest O157 contamination was observed at low strata at several sampling rounds. Persistence of STEC was not detected. Sixteen strains (28.1%) were resistant to ampicillin, streptomycin, amikacin, or tetracycline. The STEC contamination level of ground beef could vary according to the sociocultural characteristics of the population. Patricia Llorente, Laura Barnech, Kinue Irino, María Valeria Rumi, and Adriana Bentancor Copyright © 2014 Patricia Llorente et al. All rights reserved. Recombinant Dense Granular Protein (GRA5) for Detection of Human Toxoplasmosis by Western Blot Thu, 29 May 2014 11:55:21 +0000 Toxoplasma gondii infects all warm-blooded animals, including humans, causing serious public health problems and great economic loss for the food industry. Commonly used serological tests require costly and hazardous preparation of whole Toxoplasma lysate antigens from tachyzoites. Here, we have evaluated an alternative method for antigen production, which involved a prokaryotic expression system. Specifically, we expressed T. gondii dense granular protein-5 (GRA5) in Escherichia coli and isolated it by affinity purification. The serodiagnostic potential of the purified recombinant GRA5 (rGRA5) was tested through Western blot analysis against 212 human patient serum samples. We found that rGRA5 protein was 100% specific for analysis of toxoplasmosis-negative human sera. Also, rGRA5 was able to detect acute and chronic T. gondii infections (sensitivities of 46.8% and 61.2%, resp.). Xiao Teng Ching, Yee Ling Lau, Mun Yik Fong, Veeranoot Nissapatorn, and Hemah Andiappan Copyright © 2014 Xiao Teng Ching et al. All rights reserved. Added Value of QuantiFERON TB-Gold in-Tube for Detecting Latent Tuberculosis Infection among Persons Living with HIV/AIDS Thu, 29 May 2014 09:34:36 +0000 Objective. To evaluate the added value of QuantiFERON TB-Gold in-Tube (QTF-GIT) over the tuberculin skin testing (TST) for detecting latent tuberculosis (TB) infection (LTBI) among patients with AIDS in a city with a low TB incidence rate (11.1/100,000 inhabitants) and universal BCG coverage. Methods. Three hundred consecutive patients with AIDS in eight outpatient sexually transmitted disease public clinics in Brasilia were submitted to QFT-IT and TST between May 2011 and March 2013. A positive result of either test was considered to be LTBI. Results. Median CD4-cell count was 477.5 cells/mm3; 295 (98.3%) were using antiretroviral therapy. Eighteen patients (6%, 95% CI: 3.6%–9.3%) had LTBI, of whom 4 (1.3%, 95% CI: 0.04%–2.63%) had only a positive TST, 8 (2.7%, 95% CI: 0.8%–4.5%) had only a QFT-GIT positive test, and 6 (2%, 95% CI: 0.4%–3.6%) had positive results for both tests. This represents an 81.8% relative increase in LTBI detection when QFT-GIT is added to TST. The concordance between both tests was 96% (). Conclusions. The QFT-GIT alone was more effective to detect LTBI than TST alone and had an 81% added value as an add-on sequential test in this population with mild immunosuppression. The cost-effectiveness of these strategies remains to be evaluated. Josiane Maria Oliveira Souza, Maria do Socorro Nantua Evangelista, and Anete Trajman Copyright © 2014 Josiane Maria Oliveira Souza et al. All rights reserved. HS1,2 Ig Enhancer Alleles Association to AIDS Progression in a Pediatric Cohort Infected with a Monophyletic HIV-Strain Tue, 20 May 2014 12:53:20 +0000 Alteration in the humoral immune response has been observed during HIV infection. The polymorphisms of enhancer HS1,2, member of the 3′ regulatory region of the Ig heavy chain cluster, may play a role in the variation of the humoral response leading to pathological conditions. To assess the role of the HS1,2 polymorphic variants in the progression of AIDS, the HS1,2-A allelic frequencies were investigated in a cohort of HIV infected pediatric subjects from a nosocomial outbreak with a monophyletic strain of HIV. From a total group of 418 HIV infected children in the outbreak cohort, 42 nonprogressors and 31 progressors without bias due to antiretroviral therapy were evaluated. HS1,2 allele *1 has been associated with nonprogressors (allelic frequency: 51.19% versus 33.87% in progressors, OR 0.5, and , while allele *2 has been associated with progression (allelic frequency: 48.39% versus 30.95% in nonprogressors, OR 2.1, and . Further, only subjects carrying allele *2 in absence of allele *1, either in homozygous condition for allele *2 [nonprogressors 2/42 (4.76%), Progressors 7/31 (22.58%), OR 5.8, and or in combination with other allelic variants [nonprogressors 7/42 (16.67%), Progressors 13/31 (41.93%), OR 3.61, and , have been associated with HIV progression to AIDS. In conclusion, while the HS1,2 allele *1 has a protective effect on HIV progression when present, allele *2 is associated with progression toward AIDS when allele *1 is absent. Carla Montesano, Vincenzo Giambra, Domenico Frezza, Paolo Palma, Eliseo Serone, Guido Castelli Gattinara, Maurizio Mattei, Giorgio Mancino, Vittorio Colizzi, and Massimo Amicosante Copyright © 2014 Carla Montesano et al. All rights reserved. Characterization of a Mycobacterium avium subsp. avium Operon Associated with Virulence and Drug Detoxification Tue, 20 May 2014 00:00:00 +0000 The lprG-p55 operon of Mycobacterium tuberculosis and Mycobacterium bovis is involved in the transport of toxic compounds. P55 is an efflux pump that provides resistance to several drugs, while LprG is a lipoprotein that modulates the host's immune response against mycobacteria. The knockout mutation of this operon severely reduces the replication of both mycobacterial species during infection in mice and increases susceptibility to toxic compounds. In order to gain insight into the function of LprG in the Mycobacterium avium complex, in this study, we assayed the effect of the deletion of lprG gene in the D4ER strain of Mycobacterium avium subsp. avium. The replacement of lprG gene with a hygromycin cassette caused a polar effect on the expression of p55. Also, a twofold decrease in ethidium bromide susceptibility was observed and the resistance to the antibiotics rifampicin, amikacin, linezolid, and rifabutin was impaired in the mutant strain. In addition, the mutation decreased the virulence of the bacteria in macrophages in vitro and in a mice model in vivo. These findings clearly indicate that functional LprG and P55 are necessary for the correct transport of toxic compounds and for the survival of MAA in vitro and in vivo. Mariana Noelia Viale, Kun Taek Park, Belén Imperiale, Andrea Karina Gioffre, María Alejandra Colombatti Olivieri, Roberto Damián Moyano, Nora Morcillo, María de la Paz Santangelo, William Davis, and María Isabel Romano Copyright © 2014 Mariana Noelia Viale et al. All rights reserved. Differential Macrophage Response to Slow- and Fast-Growing Pathogenic Mycobacteria Sun, 18 May 2014 12:50:16 +0000 Nontuberculous mycobacteria (NTM) have recently been recognized as important species that cause disease even in immunocompetent individuals. The mechanisms that these species use to infect and persist inside macrophages are not well characterised. To gain insight concerning this process we used THP-1 macrophages infected with M. abscessus, M. fortuitum, M. celatum, and M. tuberculosis. Our results showed that slow-growing mycobacteria gained entrance into these cells with more efficiency than fast-growing mycobacteria. We have also demonstrated that viable slow-growing M. celatum persisted inside macrophages without causing cell damage and without inducing reactive oxygen species (ROS), as M. tuberculosis caused. In contrast, fast-growing mycobacteria destroyed the cells and induced high levels of ROS. Additionally, the macrophage cytokine pattern induced by M. celatum was different from the one induced by either M. tuberculosis or fast-growing mycobacteria. Our results also suggest that, in some cases, the intracellular survival of mycobacteria and the immune response that they induce in macrophages could be related to their growth rate. In addition, the modulation of macrophage cytokine production, caused by M. celatum, might be a novel immune-evasion strategy used to survive inside macrophages that is different from the one reported for M. tuberculosis. A. Cecilia Helguera-Repetto, Rommel Chacon-Salinas, Jorge F. Cerna-Cortes, Sandra Rivera-Gutierrez, Vianney Ortiz-Navarrete, Iris Estrada-Garcia, and Jorge A. Gonzalez-y-Merchand Copyright © 2014 A. Cecilia Helguera-Repetto et al. All rights reserved. Microbubble-Mediated Ultrasound Enhances the Lethal Effect of Gentamicin on Planktonic Escherichia coli Thu, 15 May 2014 14:21:59 +0000 Previous research has found that low-intensity ultrasound enhanced the lethal effect of gentamicin on planktonic E. coli. We aimed to further investigate whether microbubble-mediated low-intensity ultrasound could further enhance the antimicrobial efficacy of gentamicin. The planktonic E. coli (ATCC 25922) was distributed to four different interventions: control (), microbubble only (), ultrasound only (), and microbubble-mediated ultrasound (). Ultrasound was applied with 100 mW/cm2 (average intensity) and 46.5 KHz, which presented no bactericidal activity. After 12 h, plate counting was used to estimate the number of bacteria, and bacterial micromorphology was observed with transmission electron microscope. The results showed that the viable counts of E. coli in were decreased by 1.01 to 1.42 log10 CFU/mL compared with (). The minimal inhibitory concentration (MIC) of gentamicin against E. coli was 1 μg/mL in the and groups, lower than that in the and groups (2 μg/mL). Transmission electron microscopy (TEM) images exhibited more destruction and higher thickness of bacterial cell membranes in the than those in other groups. The reason might be the increased permeability of cell membranes for gentamicin caused by acoustic cavitation. Han-Xiao Zhu, Xun-Zi Cai, Zhong-Li Shi, Bin Hu, and Shi-Gui Yan Copyright © 2014 Han-Xiao Zhu et al. All rights reserved. Chagas’ Disease: Pregnancy and Congenital Transmission Thu, 15 May 2014 07:40:07 +0000 Chagas disease is a chronic infection that kills approximately 12,000 people a year. Mass migration of chronically infected and asymptomatic persons has caused globalization of Chagas disease and has made nonvectorial infection, including vertical and blood-borne transmission, more of a threat to human communities than vectorial infection. To control transmission, it is essential to test all pregnant women living in endemic countries and all pregnant women having migrated from, or having lived in, endemic countries. All children born to seropositive mothers should be tested not only within the first month of life but also at ~6 months and ~12 months of age. The diagnosis is made by identification of the parasite in blood before the age of 6 months and by identification of the parasite in blood and/or positive serology after 10 months of age. Follow up for a year is essential as a significant proportion of cases are initially negative and are only detected at a later stage. If the condition is diagnosed and treated early, the clinical response is excellent and the majority of cases are cured. Ana María Cevallos and Roberto Hernández Copyright © 2014 Ana María Cevallos and Roberto Hernández. All rights reserved. Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis Infection Tue, 13 May 2014 12:01:58 +0000 This study examined the immunogenic properties of the fusion protein fimbria 2 of Bordetella pertussis (Fim2)—cholera toxin B subunit (CTB) in the intranasal murine model of infection. To this end B. pertussis Fim2 coding sequence was cloned downstream of the cholera toxin B subunit coding sequence. The expression and assembly of the fusion protein into pentameric structures (CTB-Fim2) were evaluated by SDS-PAGE and monosialotetrahexosylgaglioside (GM1-ganglioside) enzyme-linked immunosorbent assay (ELISA). To evaluate the protective capacity of CTB-Fim2, an intraperitoneal or intranasal mouse immunization schedule was performed with 50 μg of CTB-Fim2. Recombinant (rFim2) or purified (BpFim2) Fim2, CTB, and phosphate-buffered saline (PBS) were used as controls. The results showed that mice immunized with BpFim2 or CTB-Fim2 intraperitoneally or intranasally presented a significant reduction in bacterial lung counts compared to control groups ( or resp.). Moreover, intranasal immunization with CTB-Fim2 induced significant levels of Fim2-specific IgG in serum and bronchoalveolar lavage (BAL) and Fim2-specific IgA in BAL. Analysis of IgG isotypes and cytokines mRNA levels showed that CTB-Fim2 results in a mixed Th1/Th2 (T-helper) response. The data presented here provide support for CTB-Fim2 as a promising recombinant antigen against Bordetella pertussis infection. Noelia Olivera, Celina E. Castuma, Daniela Hozbor, María E. Gaillard, Martín Rumbo, and Ricardo M. Gómez Copyright © 2014 Noelia Olivera et al. All rights reserved. Naturally Occurring Compounds Elicit HIV-1 Replication in Chronically Infected Promonocytic Cells Mon, 12 May 2014 07:18:07 +0000 Since antiretroviral therapy suppresses but does not eradicate HIV-1 infection, methods to purge viral reservoirs are required. Many strategies involve the reactivation of chronically HIV infected cells to induce the expression of integrated viral genome. In this study, five bioactive compounds, the plant derivatives 1-cinnamoyl-3,11-dihydroxymeliacarpin (CDM), nordihydroguaiaretic acid (NDGA), and curcumin (Cur) and the synthetic stigmasterol analogs (22S,23S)-22,23-dihydroxystigmast-4-en-3-one (compound 1) and (22S,23S)-3β-bromo-5α,22,23-trihydroxystigmastan-6-one (compound 2), were evaluated for their ability to elicit HIV replication in promonocytic (U1) and lymphocytic (H9+) HIV-1 chronically infected cells. The results revealed that natural compounds CDM, NDGA, and Cur were able to increase HIV-1 p24 antigen, determined by ELISA, only in latently infected promonocytic cells. CDM would reactivate HIV from latency by modulating the release of IL-6 and TNF-α, since the amount of both cytokines measured through ELISA significantly increased in U1 treated cells. Besides, NDGA increased ROS production, which might be related to the increase on p24 level observed in NDGA treated U1. These findings suggest that CDM, NDGA, and Cur might be candidates for further studies on latency-reversing therapeutics to eliminate latently HIV-1 reservoirs. Andrea Alejandra Barquero, María Eugenia Dávola, Diego Ariel Riva, Susana Esther Mersich, and Laura Edith Alché Copyright © 2014 Andrea Alejandra Barquero et al. All rights reserved. Atypical Enteropathogenic Escherichia coli Secretes Plasmid Encoded Toxin Sun, 11 May 2014 09:33:09 +0000 Plasmid encoded toxin (Pet) is a serine protease originally described in enteroaggregative Escherichia coli (EAEC) prototype strain 042 whose entire characterization was essentially obtained from studies performed with the purified toxin. Here we show that Pet is not exclusive to EAEC. Atypical enteropathogenic Escherichia coli (aEPEC) strains, isolated from diarrhea cases, express Pet and its detection in supernatants of infected HEp-2 cells coincides with the appearance of cell damage, which, in turn, were similar to those described with purified Pet. Pet secretion and the cytotoxic effects are time and culture medium dependent. In presence of DMEM supplemented with tryptone cell rounding and detachment were observed after just 5 h of incubation with the bacteria. In the absence of tryptone, the cytotoxic effects were detected only after 24 h of infection. We also show that, in addition to the prototype EAEC, other pet+ EAEC strains, also isolated from diarrhea cases, induce cellular damage in the same degree as the aEPEC. The cytotoxic effects of EAEC and aEPEC strains were significantly reduced in the presence of a serine protease inhibitor or anti-Pet IgG serum. Our results show a common aspect between the aEPEC and EAEC and provide the first evidence pointing to a role of Pet in aEPEC pathogenesis. Rita C. Ruiz, Keyde C. M. Melo, Sarita S. Rossato, Camila M. Barbosa, Lívia M. Corrêa, Waldir P. Elias, and Roxane M. F. Piazza Copyright © 2014 Rita C. Ruiz et al. All rights reserved. Characterization of the Opp Peptide Transporter of Corynebacterium pseudotuberculosis and Its Role in Virulence and Pathogenicity Thu, 08 May 2014 12:47:49 +0000 Despite the economic importance of caseous lymphadenitis (CLA), a chronic disease caused by Corynebacterium pseudotuberculosis, few genes related to the virulence of its etiologic agent have been characterized. The oligopeptide permease (Opp) transporters are located in the plasma membrane and have functions generally related to the uptake of peptides from the extracellular environment. These peptide transporters, in addition to having an important role in cell nutrition, also participate in the regulation of various processes involving intercellular signaling, including the control of the expression of virulence genes in pathogenic bacteria. To study the role of Opp in C. pseudotuberculosis, an OppD deficient strain was constructed via simple crossover with a nonreplicative plasmid carrying part of the oppD gene sequence. As occurred to the wild-type, the ΔoppD strain showed impaired growth when exposed to the toxic glutathione peptide (GSH), indicating two possible scenarios: (i) that this component can be internalized by the bacterium through an Opp-independent pathway or (ii) that there is toxicity while the peptide is extracellular. Additionally, the ΔoppD mutant presented a reduced ability to adhere to and infect macrophages compared to the wild-type, although both strains exhibit the same potential to colonize spleens and cause injury and death to infected mice. Pablo M. R. O. Moraes, Nubia Seyffert, Wanderson M. Silva, Thiago L. P. Castro, Renata F. Silva, Danielle D. Lima, Raphael Hirata Jr., Artur Silva, Anderson Miyoshi, and Vasco Azevedo Copyright © 2014 Pablo M. R. O. Moraes et al. All rights reserved. Xanthan Gum as an Adjuvant in a Subunit Vaccine Preparation against Leptospirosis Wed, 07 May 2014 09:09:57 +0000 Leptospiral immunoglobulin-like (Lig) proteins are of great interest due to their ability to act as mediators of pathogenesis, serodiagnostic antigens, and immunogens. Purified recombinant LigA protein is the most promising subunit vaccine candidate against leptospirosis reported to date, however, as purified proteins are weak immunogens the use of a potent adjuvant is essential for the success of LigA as a subunit vaccine. In the present study, we compared xanthan pv. pruni (strain 106), aluminium hydroxide (alhydrogel), and CpG ODN as adjuvants in a LigA subunit vaccine preparation. Xanthan gum is a high molecular weight extracellular polysaccharide produced by fermentation of Xanthomonas spp., a plant-pathogenic bacterium genus. Preparations containing xanthan induced a strong antibody response comparable to that observed when alhydrogel was used. Upon challenge with a virulent strain of L. interrogans serovar Copenhageni, significant protection (Fisher test, ) was observed in 100%, 100%, and 67% of hamsters immunized with rLigANI-xanthan, LigA-CpG-xanthan, and rLigANI-alhydrogel, respectively. Furthermore, xanthan did not cause cytotoxicity in Chinese hamster ovary (CHO) cells in vitro. The use of xanthan as an adjuvant is a novel alternative for enhancing the immunogenicity of vaccines against leptospirosis and possibly against other pathogens. Katia L. Bacelo, Daiane D. Hartwig, Fabiana K. Seixas, Rodrigo Schuch, Angelita da S. Moreira, Marta Amaral, Tiago Collares, Claire T. Vendrusculo, Alan J. A. McBride, and Odir A. Dellagostin Copyright © 2014 Katia L. Bacelo et al. All rights reserved. Bacterial Sepsis in Patients with Visceral Leishmaniasis in Northwest Ethiopia Tue, 06 May 2014 13:19:39 +0000 Background and Objectives. Visceral leishmaniasis (VL) is one of the neglected diseases affecting the poorest segment of world populations. Sepsis is one of the predictors for death of patients with VL. This study aimed to assess the prevalence and factors associated with bacterial sepsis, causative agents, and their antimicrobial susceptibility patterns among patients with VL. Methods. A cross-sectional study was conducted among parasitologically confirmed VL patients suspected of sepsis admitted to the University of Gondar Hospital, Northwest Ethiopia, from February 2012 to May 2012. Blood cultures and other clinical samples were collected and cultured following the standard procedures. Results. Among 83 sepsis suspected VL patients 16 (19.3%) had culture confirmed bacterial sepsis. The most frequently isolated organism was Staphylococcus aureus (68.8%; 11/16), including two methicillin-resistant isolates (MRSA). Patients with focal bacterial infection were more likely to have bacterial sepsis (). Conclusions. The prevalence of culture confirmed bacterial sepsis was high, predominantly due to S. aureus. Concurrent focal bacterial infection was associated with bacterial sepsis, suggesting that focal infections could serve as sources for bacterial sepsis among VL patients. Careful clinical evaluation for focal infections and prompt initiation of empiric antibiotic treatment appears warranted in VL patients. Mengistu Endris, Yegnasew Takele, Desalegn Woldeyohannes, Moges Tiruneh, Rezika Mohammed, Feleke Moges, Lutgarde Lynen, Jan Jacobs, Johan van Griensven, and Ermias Diro Copyright © 2014 Mengistu Endris et al. All rights reserved. Atypical Enteropathogenic Escherichia coli Strains form Biofilm on Abiotic Surfaces Regardless of Their Adherence Pattern on Cultured Epithelial Cells Tue, 06 May 2014 13:01:01 +0000 The aim of this study was to determine the capacity of biofilm formation of atypical enteropathogenic Escherichia coli (aEPEC) strains on abiotic and biotic surfaces. Ninety-one aEPEC strains, isolated from feces of children with diarrhea, were analyzed by the crystal violet (CV) assay on an abiotic surface after 24 h of incubation. aEPEC strains representing each HEp-2 cell type of adherence were analyzed after 24 h and 6, 12, and 18 days of incubation at 37°C on abiotic and cell surfaces by CFU/cm2 counting and confocal laser scanning microscopy (CLSM). Biofilm formation on abiotic surfaces occurred in 55 (60.4%) of the aEPEC strains. There was no significant difference in biofilm biomass formation on an abiotic versus prefixed cell surface. The biofilms could be visualized by CLSM at various developmental stages. aEPEC strains are able to form biofilm on an abiotic surface with no association with their adherence pattern on HEp-2 cells with the exception of the strains expressing UND (undetermined adherence). This study revealed the capacity of adhesion and biofilm formation by aEPEC strains on abiotic and biotic surfaces, possibly playing a role in pathogenesis, mainly in cases of persistent diarrhea. Hebert F. Culler, Cristiane M. Mota, Cecilia M. Abe, Waldir P. Elias, Marcelo P. Sircili, and Marcia R. Franzolin Copyright © 2014 Hebert F. Culler et al. All rights reserved. Effects of Shiga Toxin Type 2 on Maternal and Fetal Status in Rats in the Early Stage of Pregnancy Mon, 05 May 2014 14:32:30 +0000 Shiga toxin type 2 (Stx2), a toxin secreted by Shiga toxin-producing Escherichia coli (STEC), could be one of the causes of maternal and fetal morbimortality not yet investigated. In this study, we examined the effects of Stx2 in rats in the early stage of pregnancy. Sprague-Dawley pregnant rats were intraperitoneally (i.p.) injected with sublethal doses of Stx2, 0.25 and 0.5 ng Stx2/g of body weight (bwt), at day 8 of gestation (early postimplantation period of gestation). Maternal weight loss and food and water intake were analyzed after Stx2 injection. Another group of rats were euthanized and uteri were collected at different times to evaluate fetal status. Immunolocalization of Stx2 in uterus and maternal kidneys was analyzed by immunohistochemistry. The presence of Stx2 receptor (globotriaosylceramide, Gb3) in the uteroplacental unit was observed by thin layer chromatography (TLC). Sublethal doses of Stx2 in rats caused maternal weight loss and pregnancy loss. Stx2 and Gb3 receptor were localized in decidual tissues. Stx2 was also immunolocalized in renal tissues. Our results demonstrate that Stx2 leads to pregnancy loss and maternal morbidity in rats in the early stage of pregnancy. This study highlights the possibility of human pregnancy loss and maternal morbidity mediated by Stx2. Flavia Sacerdoti, María M. Amaral, Elsa Zotta, Ana M. Franchi, and Cristina Ibarra Copyright © 2014 Flavia Sacerdoti et al. All rights reserved. Classification of Epidemic Community-Acquired Methicillin-Resistant Staphylococcus aureus by Anatomical Site of Isolation Mon, 05 May 2014 14:16:36 +0000 Methicillin-resistant Staphylococcus aureus contributes significantly to cost, morbidity, and mortality due to infectious disease. We surveyed community-associated MRSA isolates to determine which strains were present within anatomical sites of interest. The most likely sources of MRSA among anatomic sites swabbed were wounds followed by the nasal cavity. The USA 300 MRSA strain was most commonly isolated among wound infections while nasal swabs largely yielded USA 100 MRSA. The frequency of isolation of USA 100 amongst community-associated strains is clinically significant as this strain is often correlated with invasive disease, exhibits broad antibiotic resistance, and has been considered to be hospital associated. The potential of USA 100 to cause serious disease and the frequency of its isolation suggest an important reservoir for opportunistic infection. These data demonstrate that MRSA epidemic clones are widespread among the community. Jill C. Roberts Copyright © 2014 Jill C. Roberts. All rights reserved. Identification of Virulence Factors Genes in Escherichia coli Isolates from Women with Urinary Tract Infection in Mexico Mon, 05 May 2014 13:43:46 +0000 E coli isolates (108) from Mexican women, clinically diagnosed with urinary tract infection, were screened to identify virulence genes, phylogenetic groups, and antibiotic resistance. Isolates were identified by MicroScan4 system; additionally, the minimum inhibitory concentration (MIC) was assessed. The phylogenetic groups and 16 virulence genes encoding adhesins, toxins, siderophores, lipopolysaccharide (LPS), and invasins were identified by PCR. Phylogenetic groups distribution was as follows: B1 9.3%, A 30.6%, B2 55.6%, and D 4.6%. Virulence genes prevalence was ecp 98.1%, fimH 86.1%, traT 77.8%, sfa/focDE 74.1%, papC 62%, iutA 48.1%, fyuA 44.4%, focG 2.8%, sfaS 1.9%, hlyA 7.4%, cnf-1 6.5%, cdt-B 0.9%, cvaC 2.8%, ibeA 2.8%, and rfc 0.9%. Regarding antimicrobial resistance it was above 50% to ampicillin/sulbactam, ampicillin, piperacillin, trimethoprim/sulfamethoxazole, ciprofloxacin, and levofloxacin. Uropathogenic E. coli clustered mainly in the pathogenic phylogenetic group B2. The isolates showed a high presence of siderophores and adhesion genes and a low presence of genes encoding toxins. The high frequency of papC gene suggests that these isolates have the ability to colonize the kidneys. High resistance to drugs considered as first choice treatment such as trimethoprim/sulfamethoxazole and fluoroquinolones was consistently observed. Daniela A. López-Banda, Erika M. Carrillo-Casas, Margarita Leyva-Leyva, Gabriel Orozco-Hoyuela, Ángel H. Manjarrez-Hernández, Sara Arroyo-Escalante, David Moncada-Barrón, Silvia Villanueva-Recillas, Juan Xicohtencatl-Cortes, and Rigoberto Hernández-Castro Copyright © 2014 Daniela A. López-Banda et al. All rights reserved. Phenotypic and Genotypic Characterization of Enterotoxigenic Escherichia coli Clinical Isolates from Northern Colombia, South America Wed, 30 Apr 2014 13:26:27 +0000 Enterotoxigenic Escherichia coli (ETEC) are major causes of childhood diarrhea in low and middle income countries including Colombia, South America. To understand the diversity of ETEC strains in the region, clinical isolates obtained from northern Colombia children were evaluated for multiple locus sequencing typing, serotyping, classical and nonclassical virulence genes, and antibiotic susceptibility. Among 40 ETEC clinical isolates evaluated, 21 (52.5%) were positive for LT gene, 13 (32.5%) for ST gene, and 6 (15%) for both ST and LT. The most prevalent colonization surface antigens (CS) were CS21 and CFA/I identified in 21 (50%) and 13 (32.5%) isolates, respectively. The eatA, irp2, and fyuA were the most common nonclassical virulence genes present in more than 60% of the isolates. Ampicillin resistance (80% of the strains) was the most frequent phenotype among ETEC strains followed by trimethoprim-sulfamethoxazole resistance (52.5%). Based on multiple locus sequencing typing (MLST), we recognize that 6 clonal groups of ETEC clinical isolates circulate in Colombia. ETEC clinical isolates from children in northern Colombia are highly diverse, yet some isolates circulating in the community belong to well-defined clonal groups that share a unique set of virulence factors, serotypes, and MLST sequence types. Julio A. Guerra, Yesenia C. Romero-Herazo, Octavio Arzuza, and Oscar G. Gómez-Duarte Copyright © 2014 Julio A. Guerra et al. All rights reserved. Falciparum Malaria as an Emerging Cause of Fever in Adults Living in Gabon, Central Africa Wed, 30 Apr 2014 06:36:25 +0000 Following the observed increase of malaria prevalence among older children in Gabon, a descriptive observational study was carried out in 2012 to determine the prevalence of malaria in adults presenting with fever in two health centres of Libreville, the capital city of Gabon. Thick- and thin-blood smears for malaria diagnosis were performed in febrile individuals aged more than 15 years old. Age, use of bed nets, previous antimalarial drug treatment, clinical symptoms, chest radiography results, and available haemoglobin data were also recorded. Among the 304 patients screened, the global malaria frequency was of 42.1% . Plasmodium (P). falciparum was the only species identified. The proportion of patients with a clinical malaria requiring parenteral treatment was 38.5%, whereas 47.5% of outpatients had uncomplicated malaria. According to WHO classification, 14 (19.7%) infected patients had severe malaria; neurological and respiratory symptoms tended to be more frequent in case of P. falciparum infection. Anaemia was found in 51.5% adults and none had severe anaemia. Almost half of adults consulting for fever in two health centres of the urban city of Libreville have malaria. The use of insecticide-treated bed nets, the screening, and the treatment of individuals with P. falciparum microscopic and submicroscopic asymptomatic infection or clinical malaria should be emphasized to reduce the transmission. Marielle K. Bouyou-Akotet, Christelle L. Offouga, Denise P. Mawili-Mboumba, Laurence Essola, Blondel Madoungou, and Maryvonne Kombila Copyright © 2014 Marielle K. Bouyou-Akotet et al. All rights reserved. Rural Residents in China Are at Increased Risk of Exposure to Tick-Borne Pathogens Anaplasma phagocytophilum and Ehrlichia chaffeensis Wed, 30 Apr 2014 00:00:00 +0000 As emerging tick born rickettsial diseases caused by A. phagocytophilum and E. chaffeensis, anaplasmosis and ehrlichiosis have become a serious threat to human and animal health throughout the world. In particular, in China, an unusual transmission of nosocomial cases of human granulocytic anaplasmosis occurred in Anhui Province in 2006 and more recent coinfection case of A. phagocytophilum and E. chaffeensis was documented in Shandong Province. Although the seroprevalence of human granulocytic anaplasmosis (former human granulocytic ehrlichiosis, HGE) has been documented in several studies, these data existed on local investigations, and also little data was reported on the seroprevalence of human monocytic ehrlichiosis (HME) in China. In this cross-sectional epidemiological study, indirect immunofluorescence antibody assay (IFA) proposed by WHO was used to detect A. phagocytophilum and E. chaffeensis IgG antibodies for 7,322 serum samples from agrarian residents from 9 provinces/cities and 819 urban residents from 2 provinces. Our data showed that farmers were at substantially increased risk of exposure. However, even among urban residents, risk was considerable. Seroprevalence of HGA and HME occurred in diverse regions of the country and tended to be the highest in young adults. Many species of ticks were confirmed carrying A. phagocytophilum organisms in China while several kinds of domestic animals including dog, goats, sheep, cattle, horse, wild rabbit, and some small wild rodents were proposed to be the reservoir hosts of A. phagocytophilum. The broad distribution of vector and hosts of the A. phagocytophilum and E. chaffeensis, especially the relationship between the generalized susceptibility of vectors and reservoirs and the severity of the disease’s clinical manifestations and the genetic variation of Chinese HGA isolates in China, is urgently needed to be further investigated. Lijuan Zhang, Hong Liu, Bianli Xu, Zhilun Zhang, Yuming Jin, Weiming Li, Qunying Lu, Liang Li, Litao Chang, Xiuchun Zhang, Desheng Fan, Minghua Cao, Manli Bao, Ying Zhang, Zengzhi Guan, Xueqin Cheng, Lina Tian, Shiwen Wang, Huilan Yu, Qiang Yu, Yong Wang, Yonggen Zhang, Xiaoyan Tang, Jieying Yin, Shijun Lao, Bin Wu, Juan Li, Weihong Li, Qiyi Xu, Yonglin Shi, and Fang Huang Copyright © 2014 Lijuan Zhang et al. All rights reserved. Virulence Factors Associated with Pediatric Shigellosis in Brazilian Amazon Tue, 29 Apr 2014 07:41:43 +0000 Shigellosis is a global human health problem and the incidence is highest among children. In the present work, main Shigella virulence genes was examined by PCR and compared to symptoms of pediatric shigellosis. Thirty Shigella isolates were identified from an etiologic study at which 1,339 children ranging 0–10 years old were enrolled. S. flexneri was the most frequent species reaching 60.0% of isolates, 22.2% were S. sonnei, and 6.6% were both S. dysenteriae and S. boydii. All Shigella infected children had diarrhea, but not all were accompanied by others symptoms of bacillary dysentery. Among major virulence genes, the PCR typing revealed ipaBCD was present in all isolates, followed by IpaH7.8, set-1A, set-1B, sen/ospD3, virF, and invE. The pathogenic potential of the ShET-1B subunit was observed in relation to dehydration () and ShET-2 related to the intestinal injury () evidenced by the presence of bloody diarrhea. Our results show associations among symptoms of shigellosis and virulence genes of clinical isolates of Shigella spp. Carolinie Batista Nobre da Cruz, Maria Carolina Scheffer de Souza, Paula Taquita Serra, Ivanildes Santos, Antonio Balieiro, Fabio Alessandro Pieri, Paulo Afonso Nogueira, and Patrícia Puccinelli Orlandi Copyright © 2014 Carolinie Batista Nobre da Cruz et al. All rights reserved. Analysis of the Virulence of an Atypical Enteropathogenic Escherichia coli Strain In Vitro and In Vivo and the Influence of Type Three Secretion System Mon, 28 Apr 2014 08:40:20 +0000 Atypical enteropathogenic Escherichia coli (aEPEC) inject various effectors into intestinal cells through a type three secretion system (T3SS), causing attaching and effacing (A/E) lesions. We investigated the role of T3SS in the ability of the aEPEC 1711-4 strain to interact with enterocytes in vitro (Caco-2 cells) and in vivo (rabbit ileal loops) and to translocate the rat intestinal mucosa in vivo. A T3SS isogenic mutant strain was constructed, which showed marked reduction in the ability to associate and invade but not to persist inside Caco-2 cells. After rabbit infection, only aEPEC 1711-4 was detected inside enterocytes at 8 and 24 hours pointing to a T3SS-dependent invasive potential in vivo. In contrast to aEPEC 1711-4, the T3SS-deficient strain no longer produced A/E lesions or induced macrophage infiltration. We also demonstrated that the ability of aEPEC 1711-4 to translocate through mesenteric lymph nodes to spleen and liver in a rat model depends on a functional T3SS, since a decreased number of T3SS mutant bacteria were recovered from extraintestinal sites. These findings indicate that the full virulence potential of aEPEC 1711-4 depends on a functional T3SS, which contributes to efficient adhesion/invasion in vitro and in vivo and to bacterial translocation to extraintestinal sites. Suely C. F. Sampaio, Fabiana C. Moreira, Ana M. A. Liberatore, Mônica A. M. Vieira, Terezinha Knobl, Fabiano T. Romão, Rodrigo T. Hernandes, Claudete S. A. Ferreira, Antônio P. Ferreira, Aloísio Felipe-Silva, Rita Sinigaglia-Coimbra, Ivan H. J. Koh, and Tania A. T. Gomes Copyright © 2014 Suely C. F. Sampaio et al. All rights reserved. Variations in KIR Genes: A Study in HIV-1 Serodiscordant Couples Sun, 27 Apr 2014 13:37:12 +0000 Background. NK cells have anti-HIV activity mediated through killer cell immunoglobulin-like receptors (KIRs). The current prospective cohort study evaluated whether variation in KIR genes is associated with HIV infection in discordant couples (DCs), where one spouse remains seronegative (HSN) despite repeated exposure to the HIV. Methods. KIR was genotyped using PCR SSP. Viral load and CD4 counts were estimated using commercially available reagents. Data were analyzed using SPSS software. Results. Among the 47 DCs, HSN spouses had significantly () higher frequencies of KIR3DS1. Regression analysis revealed significant () association of KIR2DS1 with low viral load. KIR2DS4 variant was associated () with high viral load. Three pairs of KIR genes were in strong LD in HSNs and two pairs in HSPs. There were 60 KIR genotypes, and 16 are reported the first time in the Indian population. Exclusive genotypes were present either in HSPs (, 11 unique genotypes) or in HSNs (, 9 unique genotypes). Conclusions. This study highlights for the first time in the Indian population an association of KIR genes in HIV infection where presence of exclusive and unique genotypes indicates possible association with either HIV infection or with protection. Vijay R. Chavan, Deepali Chaudhari, Swati Ahir, Zakiya Ansari, Preeti Mehta, and Jayanti Mania-Pramanik Copyright © 2014 Vijay R. Chavan et al. All rights reserved.