Biochemistry Research International The latest articles from Hindawi Publishing Corporation © 2016 , Hindawi Publishing Corporation . All rights reserved. Crystal Structures of Two Isozymes of Citrate Synthase from Sulfolobus tokodaii Strain 7 Tue, 30 Aug 2016 13:24:52 +0000 Thermoacidophilic archaeon Sulfolobus tokodaii strain 7 has two citrate synthase genes (ST1805-CS and ST0587-CS) in the genome with 45% sequence identity. Because they exhibit similar optimal temperatures of catalytic activity and thermal inactivation profiles, we performed structural comparisons between these isozymes to elucidate adaptation mechanisms to high temperatures in thermophilic CSs. The crystal structures of ST1805-CS and ST0587-CS were determined at 2.0 Å and 2.7 Å resolutions, respectively. Structural comparison reveals that both of them are dimeric enzymes composed of two identical subunits, and these dimeric structures are quite similar to those of citrate synthases from archaea and eubacteria. ST0587-CS has, however, 55 ion pairs within whole dimer structure, while having only 36 in ST1805-CS. Although the number and distributions of ion pairs are distinct from each other, intersubunit ion pairs between two domains of each isozyme are identical especially in interterminal region. Because the location and number of ion pairs are in a trend with other CSs from thermophilic microorganisms, the factors responsible for thermal adaptation of ST-CS isozymes are characterized by ion pairs in interterminal region. Midori Murakami and Tsutomu Kouyama Copyright © 2016 Midori Murakami and Tsutomu Kouyama. All rights reserved. Presynaptic Proteins as Markers of the Neurotoxic Activity of BmjeTX-I and BmjeTX-II Toxins from Bothrops marajoensis (Marajó Lancehead) Snake Venom Thu, 18 Aug 2016 16:31:56 +0000 Neuromuscular preparations exposed to B. marajoensis venom show increases in the frequency of miniature end-plate potentials and twitch tension facilitation followed by presynaptic neuromuscular paralysis, without evidences of muscle damage. Considering that presynaptic toxins interfere into the machinery involved in neurotransmitter release (synaptophysin, synaptobrevin, and SNAP25 proteins), the main objective of this communication is to analyze, by immunofluorescence and western blotting, the expression of the synaptic proteins, synaptophysin, synaptobrevin, and SNAP25 and by myography, light, and transmission electron microscopy the pathology of motor nerve terminals and skeletal muscle fibres of chick biventer cervicis preparations (CBC) exposed in vitro to BmjeTX-I and BmjeTX-II toxins from B. marajoensis venom. CBC incubated with toxins showed irreversible twitch tension blockade and unaffected KCl- and ACh-evoked contractures, and the positive colabelling of acetylcholine receptors confirmed that their action was primarily at the motor nerve terminal. Hypercontraction and loose myofilaments and synaptic vesicle depletion and motor nerve damage indicated that the toxins displayed both myotoxic and neurotoxic effect. The blockade resulted from interference on synaptophysin, synaptobrevin, and SNAP25 proteins leading to the conclusion that BmjeTX-I and BmjeTX-II affected neurotransmitter release machinery by preventing the docking of synaptic vesicles to the axolemma of the nerve terminal. Antonio Lisboa, Rodolfo Melaré, Junia R. B. Franco, Carolina V. Bis, Marta Gracia, Luis A. Ponce-Soto, Sérgio Marangoni, Léa Rodrigues-Simioni, Maria Alice da Cruz-Höfling, and Thalita Rocha Copyright © 2016 Antonio Lisboa et al. All rights reserved. Urinary Markers of Tubular Injury in HIV-Infected Patients Sun, 17 Jul 2016 11:26:29 +0000 Renal disease is a common complication of HIV-infected patients, associated with increased risk of cardiovascular events, progression to AIDS, AIDS-defining illness, and mortality. Early and accurate identification of renal disease is therefore crucial to improve patient outcomes. The use of serum creatinine, along with proteinuria, to detect renal involvement is essentially to screen for markers of glomerular disease and may not be effective in detecting earlier stages of renal injury. Therefore, more sensitive and specific markers are needed in order to early identify HIV-infected patients at risk of renal disease. This review article summarizes some new and important urinary markers of tubular injury in HIV-infected patients and their clinical usefulness in the renal safety follow-up of TDF-treated patients. Temesgen Fiseha and Angesom Gebreweld Copyright © 2016 Temesgen Fiseha and Angesom Gebreweld. All rights reserved. A Bioinformatics Analysis Reveals a Group of MocR Bacterial Transcriptional Regulators Linked to a Family of Genes Coding for Membrane Proteins Thu, 30 Jun 2016 09:21:15 +0000 The MocR bacterial transcriptional regulators are characterized by an N-terminal domain, 60 residues long on average, possessing the winged-helix-turn-helix (wHTH) architecture responsible for DNA recognition and binding, linked to a large C-terminal domain (350 residues on average) that is homologous to fold type-I pyridoxal 5′-phosphate (PLP) dependent enzymes like aspartate aminotransferase (AAT). These regulators are involved in the expression of genes taking part in several metabolic pathways directly or indirectly connected to PLP chemistry, many of which are still uncharacterized. A bioinformatics analysis is here reported that studied the features of a distinct group of MocR regulators predicted to be functionally linked to a family of homologous genes coding for integral membrane proteins of unknown function. This group occurs mainly in the Actinobacteria and Gammaproteobacteria phyla. An analysis of the multiple sequence alignments of their wHTH and AAT domains suggested the presence of specificity-determining positions (SDPs). Mapping of SDPs onto a homology model of the AAT domain hinted at possible structural/functional roles in effector recognition. Likewise, SDPs in wHTH domain suggested the basis of specificity of Transcription Factor Binding Site recognition. The results reported represent a framework for rational design of experiments and for bioinformatics analysis of other MocR subgroups. Teresa Milano, Sebastiana Angelaccio, Angela Tramonti, Martino Luigi Di Salvo, Roberto Contestabile, and Stefano Pascarella Copyright © 2016 Teresa Milano et al. All rights reserved. Potential Properties of Plant Sprout Extracts on Amyloid β Mon, 27 Jun 2016 12:12:03 +0000 The aim of this study is to examine the amyloid β (Aβ) inhibition mechanism of plant sprouts’ aqueous extracts (PSAE). In this study, we screened the effects of five plant sprouts’ extracts on Aβ (1–42) structure modification using gel electrophoresis. In PSAE, no band of Aβ monomer was recognized in Japanese butterbur. Similarly, the Aβ monomer band became light in buckwheat, red cabbage, broccoli, and brussels. The neuroprotective effects of PSAE were evaluated by measuring levels of Aβ in mixtures (Aβ  and PSAE) with Aβ ELISA assay. The treatment with PSAE decreased Aβ levels. The results indicated that the levels of red cabbage, Japanese butterbur, and broccoli were 9.6, 28.0, and 44.0%, respectively. The lowest value was observed with buckwheat. Furthermore, we carried out a Congo Red (CR) and Aβ binding experiment of PSAE to confirm the modification mechanism of PSAE. The correlation coefficient for the absorption spectrum peak of CR was found to be bigger than 0.8 () which proved that the Aβ levels could be attributed to the peak of CR. In conclusion, we demonstrated that treatment with PSAE effectively decreases Aβ concentration. Thus, the mechanism that decreased the Aβ levels may be modification by PSAE. Mizue Okada and Yoshinori Okada Copyright © 2016 Mizue Okada and Yoshinori Okada. All rights reserved. Cetuximab and Cisplatin Show Different Combination Effect in Nasopharyngeal Carcinoma Cells Lines via Inactivation of EGFR/AKT Signaling Pathway Tue, 24 May 2016 14:25:45 +0000 Nasopharyngeal carcinoma (NPC) is a common malignant cancer in South China. Cisplatin is a classical chemotherapeutic employed for NPC treatment. Despite the use of cisplatin-based concurrent chemoradiotherapy, distant failure still confuses clinicians and the outcome of metastatic NPC remains disappointing. Hence, a potent systemic therapy is needed for this cancer. Epidermal growth factor receptor (EGFR) represents a promising new therapeutic target in cancer. We predicted that combining the conventional cytotoxic drug cisplatin with the novel molecular-targeted agent cetuximab demonstrates a strong antitumor effect on NPC cells. In this study, we selected HNE1 and CNE2 cells, which have been proved to possess different EGFR expression levels, to validate our conjecture. The two-drug regimen showed a significant synergistic effect in HNE1 cells but an additive effect in CNE2 cells. Our results showed that cisplatin-induced apoptosis was significantly enhanced by cetuximab in the high EGFR-expressing HNE1 cells but not in CNE2 cells. Further molecular mechanism study indicated that the EGFR/AKT pathway may play an important role in cell apoptosis via the mitochondrial-mediated intrinsic pathway and lead to the different antitumor effects of this two-drug regimen between HNE1 and CNE2 cells. Thus, the regimen may be applied in personalized NPC treatments. Jiajia Gu, Li Yin, Jing Wu, Nan Zhang, Teng Huang, Kai Ding, Haixia Cao, Lin Xu, and Xia He Copyright © 2016 Jiajia Gu et al. All rights reserved. Para-Phenylenediamine Induces Apoptotic Death of Melanoma Cells and Reduces Melanoma Tumour Growth in Mice Tue, 17 May 2016 06:23:07 +0000 Melanoma is one of the most aggressive forms of cancer, usually resistant to standard chemotherapeutics. Despite a huge number of clinical trials, any success to find a chemotherapeutic agent that can effectively destroy melanoma is yet to be achieved. Para-phenylenediamine (p-PD) in the hair dyes is reported to purely serve as an external dyeing agent. Very little is known about whether p-PD has any effect on the melanin producing cells. We have demonstrated p-PD mediated apoptotic death of both human and mouse melanoma cells in vitro. Mouse melanoma tumour growth was also arrested by the apoptotic activity of intraperitoneal administration of p-PD with almost no side effects. This apoptosis is shown to occur primarily via loss of mitochondrial membrane potential (MMP), generation of reactive oxygen species (ROS), and caspase 8 activation. p-PD mediated apoptosis was also confirmed by the increase in sub-G0/G1 cell number. Thus, our experimental observation suggests that p-PD can be a potential less expensive candidate to be developed as a chemotherapeutic agent for melanoma. Debajit Bhowmick, Kaushik Bhar, Sanjaya K. Mallick, Subhadip Das, Nabanita Chatterjee, Tuhin Subhra Sarkar, Rajarshi Chakrabarti, Krishna Das Saha, and Anirban Siddhanta Copyright © 2016 Debajit Bhowmick et al. All rights reserved. Perspective of Fe3O4 Nanoparticles Role in Biomedical Applications Thu, 12 May 2016 13:32:18 +0000 In recent years, although many review articles have been presented about bioapplications of magnetic nanoparticles by some research groups with different expertise such as chemistry, biology, medicine, pharmacology, and materials science and engineering, the majority of these reviews are insufficiently comprehensive in all related topics like magnetic aspects of process. In the current review, it is attempted to carry out the inclusive surveys on importance of magnetic nanoparticles and especially magnetite ones and their required conditions for appropriate performance in bioapplications. The main attentions of this paper are focused on magnetic features which are less considered. Accordingly, the review contains essential magnetic properties and their measurement methods, synthesis techniques, surface modification processes, and applications of magnetic nanoparticles. Mohammad Reza Ghazanfari, Mehrdad Kashefi, Seyyedeh Fatemeh Shams, and Mahmoud Reza Jaafari Copyright © 2016 Mohammad Reza Ghazanfari et al. All rights reserved. Production of Laccase by Cochliobolus sp. Isolated from Plastic Dumped Soils and Their Ability to Degrade Low Molecular Weight PVC Thu, 12 May 2016 11:47:50 +0000 One of the utmost man-made problems faced today has been the ever-increasing plastic waste filling the world. It accounts for an estimated 20–30% (by volume) of municipal solid waste in landfill sites worldwide. Research on plastic biodegradation has been steadily growing over the past four decades. Several fungi have been identified that produce enzymes capable of plastic degradation in various laboratory conditions. This paper presents a study that determined the ability of fungi to degrade low molecular weight polyvinyl chloride (PVC) by the enzyme laccase. We have isolated a fungal species, Cochliobolus sp., from plastic dumped soils and they were cultured on Czapek Dox Agar slants at 30°C. The effectiveness of this fungal species on the degradation of commercial low molecular weight polyvinyl chloride (PVC) was studied under laboratory conditions. Significant differences were observed from the FTIR, GC-MS, and SEM results in between control and Cochliobolus sp. treated PVC. Tirupati Sumathi, Buddolla Viswanath, Akula Sri Lakshmi, and D. V. R. SaiGopal Copyright © 2016 Tirupati Sumathi et al. All rights reserved. Effect of pH, Temperature, and Chemicals on the Endoglucanases and β-Glucosidases from the Thermophilic Fungus Myceliophthora heterothallica F.2.1.4. Obtained by Solid-State and Submerged Cultivation Sun, 08 May 2016 09:53:11 +0000 This work reports endoglucanase and beta-glucosidase production by the thermophilic fungus Myceliophthora heterothallica in solid-state (SSC) and submerged (SmC) cultivation. Wheat bran and sugarcane bagasse were used for SSC and cardboard for SmC. Highest endoglucanase production in SSC occurred after 192 hours: 1,170.6 ± 0.8 U/g, and in SmC after 168 hours: 2,642 ± 561 U/g. The endoglucanases and beta-glucosidases produced by both cultivation systems showed slight differences concerning their optimal pH and temperature. The number of endoglucanases was also different: six isoforms in SSC and ten in SmC. Endoglucanase activity remained above 50% after incubation between pH 3.0 and 9.0 for 24 h for both cultivation systems. The effect of several chemicals displayed variation between SSC and SmC isoenzymes. Manganese activated the enzymes from SmC but inhibited those from SSC. For β-glucosidases, maximum production on SmC was 244 ± 48 U/g after 168 hours using cardboard as carbon source. In SSC maximum production reached 10.9 ± 0.3 U/g after 240 h with 1 : 1 wheat bran and sugarcane bagasse. Manganese exerted a significant activation on SSC β-glucosidases, and glucose inhibited the enzymes from both cultivation systems. FeCl3 exerted the strongest inhibition for endoglucanases and β-glucosidases. Vanessa de Cássia Teixeira da Silva, Amanda Lais de Souza Coto, Rafael de Carvalho Souza, Marcello Bertoldi Sanchez Neves, Eleni Gomes, and Gustavo Orlando Bonilla-Rodriguez Copyright © 2016 Vanessa de Cássia Teixeira da Silva et al. All rights reserved. Phytochemical Evaluation of Moth Bean (Vigna aconitifolia L.) Seeds and Their Divergence Sat, 30 Apr 2016 14:35:56 +0000 In the present study, phytochemical contents of 25 moth bean (Vigna aconitifolia) seed accessions were evaluated. This includes protease inhibitors, phytic acid, radical scavenging activity, and tannins. The studies revealed significant variation in the contents of theses phytochemicals. Presence of photochemical composition was correlated with seed storage proteins like albumin and globulin. Qualitative identification of total seed storage protein abundance across two related moth bean accessions using two-dimensional gel electrophoresis (2D-GE) was performed. Over 20 individual protein fractions were distributed over the gel as a series of spots in two moth bean accessions. Seed proteome accumulated spots of high intensity over a broad range of pI values of 3–10 in a molecular weight range of 11–170 kDa. In both seed accessions maximum protein spots are seen in the pI range of 6–8. Neha Gupta, Nidhi Shrivastava, Pramod Kumar Singh, and Sameer S. Bhagyawant Copyright © 2016 Neha Gupta et al. All rights reserved. Curcumin Mediated Attenuation of Carbofuran Induced Oxidative Stress in Rat Brain Sun, 24 Apr 2016 14:16:47 +0000 The indiscriminate use of carbofuran to improve crop productivity causes adverse effects in nontargets including mammalian systems. The objective of this study was to evaluate carbofuran induced oxidative stress in rat brain stem and its attenuation by curcumin, a herbal product. Out of 6 groups of rats, 2 groups received two different doses of carbofuran, that is, 15 and 30% of LD50, respectively, for 30 days. Out of these, 2 groups receiving same doses of carbofuran were pretreated with curcumin (100 mg/kg body weight). The levels of antioxidants, TBARS, GSH, SOD, catalase, and GST were determined in rat brain stem. The 2 remaining groups served as placebo and curcumin treated, respectively. The data suggested that carbofuran at different doses caused significant alterations in the levels of TBARS and GSH in dose dependent manner. The TBARS and GSH contents were elevated. The activities of SOD, catalase, and GST were significantly inhibited at both doses of carbofuran. The ratio of P/A was also found to be sharply increased. The pretreatment of curcumin exhibited significant protection from carbofuran induced toxicity. The results suggested that carbofuran at sublethal doses was able to induce oxidative stress in rat brain which could be attenuated by curcumin. Sunil Kumar Jaiswal, Ashish Sharma, Vivek Kumar Gupta, Rakesh Kumar Singh, and Bechan Sharma Copyright © 2016 Sunil Kumar Jaiswal et al. All rights reserved. Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes Thu, 14 Apr 2016 12:41:32 +0000 The promastigote stage of Leishmania resides in the sand fly gut, enriched with sugar molecules. Recently we reported that Leishmania donovani possesses a sucrose uptake system and a stable pool of intracellular sucrose metabolizing enzyme. In the present study, we purified the intracellular sucrase nearly to its homogeneity and compared it with the purified extracellular sucrase. The estimated size of intracellular sucrase is ~112 kDa by gel filtration chromatography, native PAGE, and substrate staining. However, in SDS-PAGE, the protein is resolved at ~56 kDa, indicating the possibility of a homodimer in its native state. The kinetics of purified intracellular sucrase shows its higher substrate affinity with a of 1.61 mM than the extracellular form having a of 4.4 mM. The highly specific activity of intracellular sucrase towards sucrose is optimal at pH 6.0 and at 30°C. In this report the purification and characterization of intracellular sucrase provide evidence that sucrase enzyme exists at least in two different forms in Leishmania donovani promastigotes. This intracellular sucrase may support further intracellular utilization of transported sucrose. Arpita Singh and Debjani Mandal Copyright © 2016 Arpita Singh and Debjani Mandal. All rights reserved. Characterization of Seed Storage Proteins from Chickpea Using 2D Electrophoresis Coupled with Mass Spectrometry Sun, 10 Apr 2016 13:13:19 +0000 Proteomic analysis was employed to map the seed storage protein network in landrace and cultivated chickpea accessions. Protein extracts were separated by two-dimensional gel electrophoresis (2D-GE) across a broad range 3.0–10.0 immobilized pH gradient (IPG) strips. Comparative elucidation of differentially expressed proteins between two diverse geographically originated chickpea accessions was carried out using 2D-GE coupled with mass spectrometry. A total of 600 protein spots were detected in these accessions. In-gel protein expression patterns revealed three protein spots as upregulated and three other as downregulated. Using trypsin in-gel digestion, these differentially expressed proteins were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) which showed 45% amino acid homology of chickpea seed storage proteins with Arabidopsis thaliana. Pramod Kumar Singh, Nidhi Shrivastava, Krishna Chaturvedi, Bechan Sharma, and Sameer S. Bhagyawant Copyright © 2016 Pramod Kumar Singh et al. All rights reserved. Potential Antioxidant Activity of New Tetracyclic and Pentacyclic Nonlinear Phenothiazine Derivatives Mon, 04 Apr 2016 11:36:15 +0000 The global increase in oxidative stress related diseases such as cancer, cardiovascular, and inflammatory diseases caused by overwhelming level of free radicals in the body has encouraged the search for new antioxidant agents. Based on the ability of newly synthesized phenothiazine derivatives (6-chloro-11-azabenzo[a]phenothiazine-5-one and 6-[4-bromophenyl]-10-methyl-11-azabenzo[a]phenothiazine-5-one) to oxidize H2O2, a known free radical to sulfoxide, this study assessed the in vitro and in vivo antioxidant activity. The synthesized phenothiazine derivatives exhibited reducing power potential to convert Fe3+ to Fe2+ and high ability to scavenge H2O2 free radical in vitro. These activities were comparable to ascorbic acid, a standard antioxidant. The catalase activity significantly increased () in groups 1 and 2 animals that received the phenothiazine derivatives compared to the controls (groups 3 and 4) suggesting the ability of the phenothiazine derivatives to scavenge H2O2 in vivo. The malondialdehyde level in groups 1 and 2 animals was lower than that in group 3 that received the reference compound (ascorbic acid) and group 4 that received the solvent suggesting the ability of the phenothiazine derivatives to prevent lipid membrane damage. AST and bilirubin levels were higher in group 2 animals which received 6-[4-bromophenyl]-10-methyl-11-azabenzo[a]phenothiazine-5-one compared to group 3, the positive control. The results suggest that phenothiazine derivatives, especially 6-chloro-11-azabenzo[a]phenothiazine-5-one, possess antioxidant activity though 6-[4-bromophenyl]-10-methyl-11-azabenzo[a]phenothiazine-5-one was slightly toxic. This activity may be due to the presence of electron donors such as sulfur as well as the richness of hydrogen in the additional benzene rings for substitution. Further study is needed to identify tolerable doses for possible therapeutic purposes. Godwill Azeh Engwa, Eugene Lekem Ayuk, Benardeth Ujunwa Igbojekwe, and Marcellus Unaegbu Copyright © 2016 Godwill Azeh Engwa et al. All rights reserved. Geometric Simulation Approach for Grading and Assessing the Thermostability of CALBs Thu, 31 Mar 2016 13:09:19 +0000 Candida antarctica lipase B (CALB) is a known stable and highly active enzyme used widely in biodiesel synthesis. In this work, the stability of native (4K6G) and mutant (4K5Q) CALB was studied through various structural parameters using conformational sampling approach. The contours of polar surface area and surface area of mutant CALB were 11357.67 Å2 and 30007.4 Å2, respectively, showing an enhanced stability compared to native CALB with a statistically significant value of < 0.0001. Moreover, simulated thermal denaturation of CALB, a process involving dilution of hydrogen bond, significantly shielded against different intervals of energy application in mutant CALB revealing its augmentation of structural rigidity against native CALB. Finally, computational docking analysis showed an increase in the binding affinity of CALB and its substrate (triglyceride) in mutant CALB with Atomic Contact Energy (ACE) of −91.23 kcal/mol compared to native CALB (ACE of −70.3 kcal/mol). The computational observations proposed that the use of mutant CALB (4K5Q) could serve as a best template for production of biodiesel in the future. Additionally, it can also be used as a template to identify efficient thermostable lipases through further mutations. B. Senthilkumar, D. Meshachpaul, and R. Rajasekaran Copyright © 2016 B. Senthilkumar et al. All rights reserved. Nationality, Gender, Age, and Body Mass Index Influences on Vitamin D Concentration among Elderly Patients and Young Iraqi and Jordanian in Jordan Thu, 24 Mar 2016 09:58:55 +0000 Vitamin D is necessary for maintaining and regulating calcium levels; thus, insufficiency of vitamin D increases the risk of many chronic diseases. This study aimed to examine vitamin D levels among Jordanian and Iraqi volunteers and find the relation between vitamin D level and lipid profile patients. Vitamin D levels were evaluated using enzyme-linked immunosorbent assay. For young healthy group subjects, vitamin D levels were 20.60 ± 5.94 ng/mL for Jordanian and 27.59 ± 7.74 ng/mL for Iraqi. Vitamin D concentrations for young males and females were 25.82 ± 8.33 ng/mL and 21.95 ± 6.39 ng/mL, respectively. Females wearing hijab were 20.87 ± 6.45 ng/mL, while uncovered females were 23.55 ± 6.04 ng/mL. For >40 years Iraqi subjects, vitamin D level for healthy was 29.78 ± 9.49 ng/mL and 23.88 ± 7.93 ng/mL for hyperlipidemic subjects. Vitamin D levels for overweight and obese healthy groups were significantly higher () than those for the hyperlipidemic patients groups. Vitamin D levels for males were significantly higher than females and were significantly higher for healthy than those hyperlipidemic Iraqi patients. These findings showed that vitamin D levels are affected by age, nationality, gender, and health statues and highlight the importance of vitamin D supplementation for groups with low levels particularly old, hijab wearing females, and hyperlipidemic groups. Hanan Al-Horani, Wael Abu Dayyih, Eyad Mallah, Mohammed Hamad, Mohammad Mima, Riad Awad, and Tawfiq Arafat Copyright © 2016 Hanan Al-Horani et al. All rights reserved. From Synthesis to Biological Impact of Pd (II) Complexes: Synthesis, Characterization, and Antimicrobial and Scavenging Activity Wed, 16 Mar 2016 12:38:21 +0000 The Pd (II) complexes with a series of halosubstituted benzylamine ligands (BLs) have been synthesized and characterized with different spectroscopic technique such as FTIR, UV/Vis, LCMS, 1H, and 13C NMR. Their molecular sustainability in different solvents such as DMSO, DMSO : H2O, and DMSO : PBS at physiological condition (pH 7.2) was determined by UV/Vis spectrophotometer. The in vitro antibacterial and antifungal activities of the complexes were investigated against Gram-positive and Gram-negative microbes and two different fungi indicated their significant biological potential. Additionally, their antioxidant activity has been analyzed with DPPH• free radical through spectrophotometric method and the result inferred them as an antioxidant. The stronger antibacterial and antioxidant activities of the synthesized complexes suggested them as a stronger antimicrobial agent. Our study advances the biological importance of palladium (II) amine complexes in the field of antimicrobial and antioxidant activities. Nitin Kumar Sharma, Rakesh Kumar Ameta, and Man Singh Copyright © 2016 Nitin Kumar Sharma et al. All rights reserved. Oxidative Stress: An Effective Prognostic Tool for an Early Detection of Cardiovascular Disease in Menopausal Women Wed, 16 Mar 2016 12:02:41 +0000 Background. Menopause, a form of reproductive aging, is marked by many hormonal variations which cause imbalance in the oxidative processes resulting in onset of endothelial dysfunction leading to cardiovascular disease (CVD). We aimed to analyze the effect of oxidative stress in an early detection of CVD in all menopausal women both normolipidemic and hyperlipidemic. Methods and Results. Study included 523 menopausal women (265 CVD and 258 non-CVD). They were screened for lipid profile, serum malondialdehyde (MDA), serum LDL carbonyl protein, and serum superoxide dismutase (SOD). Pearson’s correlation was observed between MDA and atherogenic index of plasma (AIP) in both normolipidemic (; ) and hyperlipidemic (; ) CVD group as compared to non-CVD menopausal women. Significant correlation was also observed between LDL carbonyl content and AIP in normolipidemic (; ) and hyperlipidemic (; ) CVD menopausal women as compared to non-CVD ones. Conclusion. Strong correlation between atherogenic index of plasma and oxidative stress in CVD menopausal women reveals oxidative stress as an effective prognostic tool for an early detection of cardiovascular risk. Jyot Amrita, Mridula Mahajan, A. J. S. Bhanwer, and Gurinder Mohan Copyright © 2016 Jyot Amrita et al. All rights reserved. Physical Training Status Determines Oxidative Stress and Redox Changes in Response to an Acute Aerobic Exercise Tue, 15 Mar 2016 09:35:32 +0000 Objective. To assess the influence of different physical training status on exercise-induced oxidative stress and changes in cellular redox state. Methods. Thirty male subjects participated in this study and were assigned as well-trained (WT), moderately trained (MT), and untrained (UT) groups. The levels of cortisol, creatine kinase, plasma reduced glutathione to oxidized glutathione (GSH/GSSG), cysteine/cystine (Cys/CySS), and GSH/GSSG ratio in red blood cells (RBCs) were measured immediately and 10 and 30 min after exercise. Results. Following the exercise, plasma GSH/GSSG () and Cys/CySS () were significantly reduced in all groups. Reduction in plasma GSH/GSSG ratio in all groups induced a transient shift in redox balance towards a more oxidizing environment without difference between groups (), while RBCs GSH/GSSG showed significant reduction () and elevation () in UT and MT groups, respectively. The highest level of RBCs GSH/GSSG ratio was recorded in MT group, and the lowest one was recorded in the WT group. Conclusion. Long term regular exercise training with moderate intensity shifts redox balance towards more reducing environment, versus intensive exercise training leads to more oxidizing environment and consequently development of related diseases. Farnaz Seifi-skishahr, Arsalan Damirchi, Manoochehr Farjaminezhad, and Parvin Babaei Copyright © 2016 Farnaz Seifi-skishahr et al. All rights reserved. Characterization of Jamaican Delonix regia and Cassia fistula Seed Extracts Thu, 10 Mar 2016 07:54:42 +0000 Delonix regia and Cassia fistula seed extracts were evaluated for their antioxidant activity, total phenolics, ash, zinc and fatty acid content. Fourier Transform Infrared Spectroscopy (FTIR) was utilized to assess the chemical functionalities present within the seeds. Antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Trolox equivalent antioxidant capacity (TEAC) assays. Total phenolics were determined by the Folin-Ciocalteu assay. Lipid extracts were characterized by nuclear magnetic resonance spectroscopy and gas chromatography/mass spectrometry. Zinc concentration was determined by atomic absorption spectroscopy. Extracts from the seeds of C. fistula had a higher antioxidant activity, free radical scavenging activity, and phenolic content than D. regia. FTIR revealed that the seeds are a rich source of protein with small quantities of fat. C. fistula extracts contained a higher percentage of total fat than D. regia. Palmitic acid was identified as the predominant saturated fatty acid in both extracts. Oleic acid and linoleic acid were identified in smaller quantities. Seed extracts may be considered for use in food and nutraceutical applications. Andrea Goldson Barnaby, Raymond Reid, Vaughn Rattray, Ruth Williams, and Marcel Denny Copyright © 2016 Andrea Goldson Barnaby et al. All rights reserved. HPV Infection and Prognostic Factors of Tongue Squamous Cell Carcinoma in Different Ethnic Groups from Geographically Closed Cohort in Xinjiang, China Thu, 10 Mar 2016 06:19:25 +0000 Background. The effect of HPV infection status and ethnic differences on the prognosis of tongue squamous cell carcinoma in Xinjiang presents an interesting set of conditions that has yet to be studied. Methods. A comprehensive analysis of clinical data was undertaken for a cohort consisting of 63 patients with tongue squamous cell carcinoma recruited from three ethnic groups in Xinjiang. PCR was used for the detection of HPV16 and HPV18 infections. Kaplan-Meier survival analysis was used for analyzing survival outcome in addition to the assessment of other prognostic factors. Results. The overall infection rate for HPV was 28.6% (18/63); the 5-year survival rate among the HPV-positive patients was 47.8% and 30.3% for HPV-negative patients. The survival rate for HPV-positive patients who received radiotherapy and chemotherapy was better than for those who did not receive radiotherapy and chemotherapy. N staging and HPV infection were found to be two independent and significant prognostic factors. Conclusion. HPV-positive patients with tongue squamous cell carcinoma are more sensitive to chemotherapy. Higher N staging indicates poor prognosis. Hua Zhang, Yang Zhang, Huarong Zhao, Huerxidan Niyaz, Pan Liu, Lei Zhang, Songan Zhang, Yiming Reheman, Yongxing Bao, and Xinhua Chen Copyright © 2016 Hua Zhang et al. All rights reserved. Changes in Lipid Profiles of HIV+ Adults over Nine Months at a Harare HIV Clinic: A Longitudinal Study Tue, 08 Mar 2016 11:39:04 +0000 HIV infection, together with ART, is associated with changes in biochemical, metabolic parameters and lipid profiles. The aim of this study was to compare changes in lipid profiles among HIV positive outpatients over nine months. 171 patients were investigated, 79% were ART experienced, and 82% of ART experienced patients were on NVP/EFV first line at baseline, but some patients changed ART groups over follow-up and classification was based on intent to treat. More than 60% ART naïve and ART experienced patients had some form of dyslipidemia either at baseline or at follow-up, but mean lipid values for the two groups were within normal limits. At baseline and follow-up, mean levels of TC and HDL were slightly higher in the ART experienced group. Interestingly, there was higher increase in HDL over time in the ART negative group compared to the ART positive group. There was a decrease in TC/HDL ratio in both groups over time, suggesting a reduction in calculated risk of CHD over time. HIV positive patients frequently show various forms of dyslipidemia, but there are no changes in average atherogenic lipid levels and results suggest reduced risk of CHD, mainly due to increases in HDL, after nine months of observation time. Danai Tavonga Zhou, Doreen Nehumba, Olav Oktedalen, Princess Marange, Vitaris Kodogo, Zvenyika Alfred Gomo, Tonya M. Esterhuizen, and Babill Stray-Pedersen Copyright © 2016 Danai Tavonga Zhou et al. All rights reserved. The Ginkgo biloba Extract Reverses the Renal Effects of Titanium Dioxide Nanoparticles in Adult Male Rats Thu, 03 Mar 2016 12:45:03 +0000 The Ginkgo biloba extract (GbE) is a commercial product used as a nutraceutic herbal remedy in Europe and US. It contains 27% of the polyphenols isorhamnetin, kaempferol, and quercetin, as antioxidants. We used male adult Wistar rats (200–300 g), divided into four groups: control group (treated with 5.0 mg/kg of sodium chloride, intravenous), titanium dioxide nanoparticles (TiO2-NPs) group (5.0 mg/kg, intravenous), GbE group (10 mg/kg, intraperitoneal), and GbE + TiO2-NPs group (treated 24 h before with 10 mg/kg of GbE, intraperitoneal), followed, 24 h later, by 5.0 mg/kg of TiO2-NPs intravenously. The statistical analysis was performed using Student’s -test for grouped data with ANOVA posttest. The GbE protected renal cells against the effects of TiO2-NPs because it reversed the increased activity of -glutamyltranspeptidase and the enzymatic activity of dipeptidylaminopeptidase IV at all times tested (0–5, 5–24, 24–48, and 48–72 h). Also it reversed the glucosuria, hypernatriuria, and urine osmolarity at three times tested (5–24, 24–48, and 48–72). Thus, we conclude that GbE has a beneficial activity in the cytoplasmic membranes of brush border cells on the renal tubules, against the adverse effects that can be produced by some xenobiotics in this case the TiO2-NPs, in experimental rats. Carlos Enrique Escárcega-González, Irma Guadalupe Reynoso-Andeola, Fernando Jaramillo-Juárez, Haydée Martínez-Ruvalcaba, and Francisco A. Posadas del Rio Copyright © 2016 Carlos Enrique Escárcega-González et al. All rights reserved. Biosensors in Health Care: The Milestones Achieved in Their Development towards Lab-on-Chip-Analysis Thu, 03 Mar 2016 09:11:46 +0000 Immense potentiality of biosensors in medical diagnostics has driven scientists in evolution of biosensor technologies and innovating newer tools in time. The cornerstone of the popularity of biosensors in sensing wide range of biomolecules in medical diagnostics is due to their simplicity in operation, higher sensitivity, ability to perform multiplex analysis, and capability to be integrated with different function by the same chip. There remains a huge challenge to meet the demands of performance and yield to its simplicity and affordability. Ultimate goal stands for providing point-of-care testing facility to the remote areas worldwide, particularly the developing countries. It entails continuous development in technology towards multiplexing ability, fabrication, and miniaturization of biosensor devices so that they can provide lab-on-chip-analysis systems to the community. Suprava Patel, Rachita Nanda, Sibasish Sahoo, and Eli Mohapatra Copyright © 2016 Suprava Patel et al. All rights reserved. Influenza A Virus and Influenza B Virus Can Induce Apoptosis via Intrinsic or Extrinsic Pathways and Also via NF-κB in a Time and Dose Dependent Manner Wed, 02 Mar 2016 08:31:49 +0000 Influenza viruses are able to cause annual epidemics and pandemics due to their mutation rates and reassortment capabilities leading to antigenic shifts and drifts. To identify host response to influenza A and B viruses on A549 and MDCK II cells at low and high MOIs, expressions of MxA and caspases 3, 8, and 9 and BAD, TNFα, and IκBα genes were measured in the cells supernatants. H1N1 and H3N2 prefer to initially enhance the intrinsic pathway, determined by higher caspase 9 activity in MDCK II cells compared to caspase 8 activity and vice versa in A549 cells at different MOIs, while INF B prefers extrinsic pathway in A549 cells according to significant low or undetectable caspase 9 activity and high activity of caspase 8 but also can induce intrinsic pathway in MDCK II cells as determined by significant low or undetectable activity of caspase 8 and high caspase 9 activity at different MOIs; the considerable MxA expression was found in influenza A and B viruses infected A549 and MDCK II cells at low MOIs. In conclusion, influenza A and B viruses induced extrinsic and intrinsic apoptosis in parallel, and the induction was associated with viral infection in a dose dependent manner. Ibrahim El-Sayed, Khalid Bassiouny, Aziz Nokaly, Ahmed S. Abdelghani, and Wael Roshdy Copyright © 2016 Ibrahim El-Sayed et al. All rights reserved. Fructose-Induced Carbonyl/Oxidative Stress in S. cerevisiae: Involvement of TOR Thu, 25 Feb 2016 09:42:08 +0000 The TOR (target of rapamycin) signaling pathway first described in the budding yeast Saccharomyces cerevisiae is highly conserved in eukaryotes effector of cell growth, longevity, and stress response. TOR activation by nitrogen sources, in particular amino acids, is well studied; however its interplay with carbohydrates and carbonyl stress is poorly investigated. Fructose is a more potent glycoxidation agent capable of producing greater amounts of reactive carbonyl (RCS) and oxygen species (ROS) than glucose. The increased RCS/ROS production, as a result of glycoxidation in vivo, is supposed to be involved in carbonyl/oxidative stress, metabolic disorders, and lifespan shortening of eukaryotes. In this work we aim to expand our understanding of how TOR is involved in carbonyl/oxidative stress caused by reducing monosaccharides. It was found that in fructose-grown compared with glucose-grown cells the level of carbonyl/oxidative stress markers was higher. The defects in the TOR pathway inhibited metabolic rate and suppressed generation of glycoxidation products in fructose-grown yeast. Bohdana V. Valishkevych, Ruslana A. Vasylkovska, Liudmyla M. Lozinska, and Halyna M. Semchyshyn Copyright © 2016 Bohdana V. Valishkevych et al. All rights reserved. In Silico Screening, Alanine Mutation, and DFT Approaches for Identification of NS2B/NS3 Protease Inhibitors Thu, 25 Feb 2016 07:15:37 +0000 To identify the ligand that binds to a target protein with high affinity is a nontrivial task in computer-assisted approaches. Antiviral drugs have been identified for NS2B/NS3 protease enzyme on the mechanism to cleave the viral protein using the computational tools. The consequence of the molecular docking, free energy calculations, and simulation protocols explores the better ligand. It provides in-depth structural insights with the catalytic triad of His51, Asp75, Ser135, and Gly133. The MD simulation was employed here to predict the stability of the complex. The alanine mutation has been performed and its stability was monitored by using the molecular dynamics simulation. The minimal RMSD value suggests that the derived complexes are close to equilibrium. The DFT outcome reveals that the HOMO-LUMO gap of Ligand19 is 2.86 kcal/mol. Among the considered ligands, Ligand19 shows the lowest gap and it is suggested that the HOMO of Ligand19 may transfer the electrons to the LUMO in the active regions. The calculated binding energy of Ligand19 using the DFT method is in good agreement with the docking studies. The pharmacological activity of ligand was performed and satisfies Lipinski rule of 5. Moreover, the computational results are compared with the available IC50 values of experimental results. R. Balajee, V. Srinivasadesikan, M. Sakthivadivel, and P. Gunasekaran Copyright © 2016 R. Balajee et al. All rights reserved. Production and Characterization of Highly Thermostable β-Glucosidase during the Biodegradation of Methyl Cellulose by Fusarium oxysporum Mon, 08 Feb 2016 12:06:58 +0000 Production of β-glucosidase from Fusarium oxysporum was investigated during degradation of some cellulosic substrates (Avicel, α-cellulose, carboxymethyl cellulose (CMC), and methylcellulose). Optimized production of β-glucosidase using the cellulosic substrate that supported highest yield of enzyme was examined over 192 h fermentation period and varied pH of 3.0–11.0. The β-glucosidase produced was characterized for its suitability for industrial application. Methyl cellulose supported the highest yield of β-glucosidase (177.5 U/mg) at pH 6.0 and 30°C at 96 h of fermentation with liberation of 2.121 μmol/mL glucose. The crude enzyme had optimum activity at pH 5.0 and 70°C. The enzyme was stable over broad pH range of 4.0–7.0 with relative residual activity above 60% after 180 min of incubation. β-glucosidase demonstrated high thermostability with 83% of its original activity retained at 70°C after 180 min of incubation. The activity of β-glucosidase was enhanced by Mn2+ and Fe2+ with relative activities of 167.67% and 205.56%, respectively, at 5 mM and 360% and 315%, respectively, at 10 mM. The properties shown by β-glucosidase suggest suitability of the enzyme for industrial applications in the improvement of hydrolysis of cellulosic compounds into fermentable sugars that can be used in energy generation and biofuel production. Folasade M. Olajuyigbe, Chidinma M. Nlekerem, and Olusola A. Ogunyewo Copyright © 2016 Folasade M. Olajuyigbe et al. All rights reserved. Contribution of Cell Surface Hydrophobicity in the Resistance of Staphylococcus aureus against Antimicrobial Agents Sun, 07 Feb 2016 06:50:05 +0000 Staphylococcus aureus is found in a wide variety of habitats, including human skin, where many strains are commensals that may be clinically significant or contaminants of food. To determine the physiological characteristics of resistant strain of Staphylococcus aureus against pediocin, a class IIa bacteriocin, a resistant strain was compared with wild type in order to investigate the contribution of hydrophobicity to this resistance. Additional clumping of resistant strain relative to wild type in light microscopy was considered as an elementary evidence of resistance attainment. A delay in log phase attainment was observed in resistant strain compared to the wild type strain. A significant increase in cell surface hydrophobicity was detected for resistant strain in both hexadecane and xylene indicating the contribution of cell surface hydrophobicity as adaptive reaction against antimicrobial agents. Puja Lather, A. K. Mohanty, Pankaj Jha, and Anita Kumari Garsa Copyright © 2016 Puja Lather et al. All rights reserved.