Biochemistry Research International The latest articles from Hindawi Publishing Corporation © 2015 , Hindawi Publishing Corporation . All rights reserved. Similar and Additive Effects of Ovariectomy and Diabetes on Insulin Resistance and Lipid Metabolism Thu, 05 Mar 2015 12:03:50 +0000 Type 2 diabetes mellitus (T2DM) is among the leading causes of death in postmenopausal women. The disruption of ovarian function may contribute to the incidence of T2DM. The purpose of this study was to investigate the effects of ovariectomy and T2DM on glucose and lipid homeostasis, perilipin levels in adipose tissues, as a lipolytic regulator, and levels of certain adipokines. Ovariectomized (OVX) rats were used as a model for postmenopausal women. The study was performed on sham, OVX, sham diabetic, and OVX diabetic female rats. The results indicated that ovariectomy alters adipose tissue metabolism through reducing perilipin content in white adipose tissue (WAT); however it has no effect on perilipin level in brown adipose tissue (BAT). OVX diabetic females suffer from serious metabolic disturbances, suggested by exacerbation of insulin resistance in terms of disrupted lipid profile, higher HOMA-IR, hyperinsulinemia, higher leptin, and lower adiponectin concentrations. These metabolic derangements may underlie the predisposition for cardiovascular disease in women after menopause. Therefore, for efficient treatment, the menopausal status of diabetic female should be addressed, and the order of events is of great importance because ovariectomy following development of diabetes has more serious complications compared to development of diabetes as result of menopause. Shady H. Tawfik, Bothaina F. Mahmoud, Mohamed I. Saad, Mona Shehata, Maher A. Kamel, and Madiha H. Helmy Copyright © 2015 Shady H. Tawfik et al. All rights reserved. Modulation of Adipocytokines Production and Serum NEFA Level by Metformin, Glimepiride, and Sitagliptin in HFD/STZ Diabetic Rats Sun, 01 Mar 2015 06:56:56 +0000 Type 2 diabetes mellitus (T2DM) is a group of metabolic disorders characterized by hyperglycemia owing to insulin resistance and/or insulin deficiency. Current theories of T2DM pathophysiology include a decline in β-cells function, a defect in insulin signaling pathways, and a dysregulation of secretory function of adipocytes. This study aimed to investigate the effect of different antidiabetic drugs on serum levels of certain adipocytokines and nonesterified fatty acids (NEFA) in high-fat diet (HFD)/streptozotocin- (STZ-) induced diabetic rats. All treatments significantly decreased serum NEFA level. Metformin and sitagliptin increased serum adiponectin level, whereas they decreased serum leptin level. Glimepiride showed significant decline in serum levels of both adiponectin and leptin. All treatments remarkably ameliorated insulin resistance, suggested by an improvement of glycemic control, a significant reduction in homeostasis model assessment of insulin resistance (HOMA-IR), and a correction in lipid profile. Modulation of adipocytokines production (i.e., increased serum adiponectin and decreased serum leptin) may also underlie the improvement of insulin resistance and could be a possible mechanism for the beneficial cardiovascular effects of metformin and sitagliptin. Mohamed I. Saad, Maher A. Kamel, and Mervat Y. Hanafi Copyright © 2015 Mohamed I. Saad et al. All rights reserved. Bp-13 PLA2: Purification and Neuromuscular Activity of a New Asp49 Toxin Isolated from Bothrops pauloensis Snake Venom Thu, 19 Feb 2015 12:30:58 +0000 A new PLA2 (Bp-13) was purified from Bothrops pauloensis snake venom after a single chromatographic step of RP-HPLC on μ-Bondapak C-18. Amino acid analysis showed a high content of hydrophobic and basic amino acids and 14 half-cysteine residues. The N-terminal sequence showed a high degree of homology with basic Asp49 PLA2 myotoxins from other Bothrops venoms. Bp-13 showed allosteric enzymatic behavior and maximal activity at pH 8.1, 36°–45°C. Full Bp-13 PLA2 activity required Ca2+; its PLA2 activity was inhibited by Mg2+, Mn2+, Sr2+, and Cd2+ in the presence and absence of 1 mM Ca2+. In the mouse phrenic nerve-diaphragm (PND) preparation, the time for 50% paralysis was concentration-dependent (). Both the replacement of Ca2+ by Sr2+ and temperature lowering (24°C) inhibited the Bp-13 PLA2-induced twitch-tension blockade. Bp-13 PLA2 inhibited the contractile response to direct electrical stimulation in curarized mouse PND preparation corroborating its contracture effect. In biventer cervicis preparations, Bp-13 induced irreversible twitch-tension blockade and the KCl evoked contracture was partially, but significantly, inhibited (). The main effect of this new Asp49 PLA2 of Bothrops pauloensis venom is on muscle fiber sarcolemma, with avian preparation being less responsive than rodent preparation. The study enhances biochemical and pharmacological characterization of B. pauloensis venom. Georgina Sucasaca-Monzón, Priscila Randazzo-Moura, Thalita Rocha, Frank Denis Torres-Huaco, Augusto Vilca-Quispe, Luis Alberto Ponce-Soto, Sérgio Marangoni, Maria Alice da Cruz-Höfling, and Léa Rodrigues-Simioni Copyright © 2015 Georgina Sucasaca-Monzón et al. All rights reserved. Phytochemical Analysis and Biological Activities of Cola nitida Bark Thu, 12 Feb 2015 11:51:56 +0000 Kola nut is chewed in many West African cultures and is used ceremonially. The aim of this study is to investigate some biological effects of Cola nitida’s bark after phytochemical screening. The bark was collected, dried, and then powdered for the phytochemical screening and extractions. Ethanol and ethyl acetate extracts of C. nitida were used in this study. The antibacterial activity was tested on ten reference strains and 28 meat isolated Staphylococcus strains by disc diffusion method. The antifungal activity of three fungal strains was determined on the Potato-Dextrose Agar medium mixed with the appropriate extract. The antioxidant activity was determined by DPPH and ABTS methods. Our data revealed the presence of various potent phytochemicals. For the reference and meat isolated strains, the inhibitory diameter zone was from  mm (C. albicans) to  mm (P. vulgaris). The MIC ranged from 0.312 mg/mL to 5.000 mg/mL and the MBC from 0.625 mg/mL to >20 mg/mL. The highest antifungal activity was observed with F. verticillioides and the lowest one with P. citrinum. The two extracts have an excellent reducing free radical activity. The killing effect of A. salina larvae was perceptible at 1.04 mg/mL. The purified extracts of Cola nitida’s bark can be used to hold meat products and also like phytomedicine. Durand Dah-Nouvlessounon, Hubert Adoukonou-Sagbadja, Nafan Diarrassouba, Haziz Sina, Adolphe Adjanohoun, Mariam Inoussa, Donald Akakpo, Joachim D. Gbenou, Simeon O. Kotchoni, Mamoudou H. Dicko, and Lamine Baba-Moussa Copyright © 2015 Durand Dah-Nouvlessounon et al. All rights reserved. Characterization of Protein Complexes and Subcomplexes in Protein-Protein Interaction Databases Wed, 04 Feb 2015 09:18:11 +0000 The identification and characterization of protein complexes implicated in protein-protein interaction data are crucial to the understanding of the molecular events under normal and abnormal physiological conditions. This paper provides a novel characterization of subcomplexes in protein interaction databases, stressing definition and representation issues, quantification, biological validation, network metrics, motifs, modularity, and gene ontology (GO) terms. The paper introduces the concept of “nested group” as a way to represent subcomplexes and estimates that around 15% of those nested group with the higher Jaccard index may be a result of data artifacts in protein interaction databases, while a number of them can be found in biologically important modular structures or dynamic structures. We also found that network centralities, enrichment in essential proteins, GO terms related to regulation, imperfect 5-clique motifs, and higher GO homogeneity can be used to identify proteins in nested complexes. Nazar Zaki, Elfadil A. Mohamed, and Antonio Mora Copyright © 2015 Nazar Zaki et al. All rights reserved. Protective Effect of Sodium Nitroprusside on the Rat Small Intestine Transplanted Mucosa Sun, 11 Jan 2015 08:33:58 +0000 The intestinal mucosal epithelium is extremely susceptible to even brief periods of ischemia. Mucosal barrier damage, which is associated with ischemia/reperfusion (I/R) injury and consequently bacterial translocation, remains a major obstacle for clinically successful small bowel transplantation (SBT). Previous studies have demonstrated a protective effect of nitric oxide (NO) on other transplanted organs and NO mediated intestinal protection has also been reported in vitro. The aim of this study was to evaluate the effect of sodium nitroprusside (SNP), NO donor, on graft mucosal histology and molecular markers of function after SBT in rats. We used SNP in different period of heterotopic SBT rats. The groups consisted of SBT, pre-SNP group, and post-SNP group. Interestingly, the pre-SNP graft samples exhibited less damage compared to the SBT and post-SNP samples. In addition, mucosal samples from the pre-SNP group showed higher Na+-K+-ATPase activity and higher levels of laminin expression compared to the SBT and post-SNP samples. The findings of the present study reveal that SNP given before graft ischemia/reperfusion injury has a protective effect on mucosal histology and molecular markers of function in the transplanted small intestine. Feng-Hua Chen, Ke Li, Lu Yin, Chun-Qiu Chen, Zhao-Wen Yan, and Gui-Ming Chen Copyright © 2015 Feng-Hua Chen et al. All rights reserved. O Serogroup-Specific Touchdown-Multiplex Polymerase Chain Reaction for Detection and Identification of Vibrio cholerae O1, O139, and Non-O1/Non-O139 Sun, 28 Dec 2014 07:17:50 +0000 A novel, sensitive locus-specific touchdown-multiplex polymerase chain reaction (TMPCR), which is based on two-stage amplification pertaining to multiplex PCR and conditional touchdown strategy, was used in detecting and differentiating Vibrio cholerae serogroups. A panel of molecular marker-based TMPCR method generates reproducible profiles of V. cholerae-specific (588 bp) amplicons derived from ompW gene encoding the outer membrane protein and serogroup-specific amplicons, 364 bp for the O1 and 256 bp for the O139, authentically copied from rfb genes responsible for the lipopolysaccharide biosynthesis. The TMPCR amplification efficiency yields either equally or unequally detectable duplex DNA bands of the O1 (588 and 364 bp) and O139 (588 and 256 bp) or a DNA fragment of non-O1/non-O139 (588 bp) while providing no false positive identifications using the genomic DNA templates of the other vibrios and Enterobacteriaceae. The reciprocal analysis of two-template combinations demonstrated that, using V. cholerae O1, O139, or equally mixed O1 and O139, the TMPCR had a detection limit of as low as 100 pg of the O1, O139, or non-O1/non-O139 in reactions containing unequally or equally mixed gDNAs. In addition, the O serogroup-specific TMPCR method had 100% agreement with the serotyping method when examined for the serotyped V. cholerae reference strains and those recovered from clinical samples. The potential benefit of using this TMPCR tool would augment the serotyping method used in epidemiological surveillance and monitoring of V. cholerae serogroups, O1, O139, and non-O1/non-O139 present in clinical and environmental samples. Adisak Bhumiratana, Achiraya Siriphap, Nutsarin Khamsuwan, Jednipit Borthong, Kaknokrat Chonsin, and Orasa Sutheinkul Copyright © 2014 Adisak Bhumiratana et al. All rights reserved. Effect of Inhaling Cymbopogon martinii Essential Oil and Geraniol on Serum Biochemistry Parameters and Oxidative Stress in Rats Tue, 09 Dec 2014 09:22:58 +0000 The effects of the inhalation of Cymbopogon martinii essential oil (EO) and geraniol on Wistar rats were evaluated for biochemical parameters and hepatic oxidative stress. Wistar rats were divided into three groups : G1 was control group, treated with saline solution; G2 received geraniol; and G3 received C. martinii EO by inhalation during 30 days. No significant differences were observed in glycemia and triacylglycerol levels; G2 and G3 decreased total cholesterol level. There were no differences in serum protein, urea, aspartate aminotransferase activity, and total hepatic protein. Creatinine levels increased in G2 but decreased in G3. Alanine aminotransferase activity and lipid hydroperoxide were higher in G2 than in G3. Catalase and superoxide dismutase activities were higher in G3. C. martinii EO and geraniol increased glutathione peroxidase. Oxidative stress caused by geraniol may have triggered some degree of hepatic toxicity, as verified by the increase in serum creatinine and alanine aminotransferase. Therefore, the beneficial effects of EO on oxidative stress can prevent the toxicity in the liver. This proves possible interactions between geraniol and numerous chemical compounds present in C. martinii EO. Bruna Fernanda Murbach Teles Andrade, Camila Pereira Braga, Klinsmann Carolo dos Santos, Lidiane Nunes Barbosa, Vera Lúcia Mores Rall, José Maurício Sforcin, Ana Angélica Henrique Fernandes, and Ary Fernandes Júnior Copyright © 2014 Bruna Fernanda Murbach Teles Andrade et al. All rights reserved. Antiatherosclerotic and Cardioprotective Potential of Acacia senegal Seeds in Diet-Induced Atherosclerosis in Rabbits Sun, 07 Dec 2014 00:10:20 +0000 Acacia senegal L. (Fabaceae) seeds are essential ingredient of “Pachkutta,” a specific Rajasthani traditional food. The present study explored antiatherosclerotic and cardioprotective potential of Acacia senegal seed extract, if any, in hypercholesterolemic diet-induced atherosclerosis in rabbits. Atherosclerosis in rabbits was induced by feeding normal diet supplemented with oral administration of cholesterol (500 mg/kg body weight/day mixed with coconut oil) for 15 days. Circulating total cholesterol (TC), HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C), triglycerides, and VLDL-cholesterol (VLDL-C) levels; atherogenic index (AI); cardiac lipid peroxidation (LPO); planimetric studies of aortal wall; and histopathological studies of heart, aorta, kidney, and liver were performed. Apart from reduced atherosclerotic plaques in aorta () and increased lumen volume (), administration with ethanolic extract of Acacia senegal seeds (500 mg/kg/day, p.o.) for 45 days to atherosclerotic rabbits significantly lowered serum TC, LDL-C, triglyceride, and VLDL-C levels and atherogenic index as compared to control. Atherogenic diet-induced cardiac LPO and histopathological abnormalities in aorta wall, heart, kidney, and liver were reverted to normalcy by Acacia senegal seed extract administration. The findings of the present study reveal that Acacia senegal seed extract ameliorated diet-induced atherosclerosis and could be considered as lead in the development of novel therapeutics. Heera Ram, Rameshwar Jatwa, and Ashok Purohit Copyright © 2014 Heera Ram et al. All rights reserved. Comparative Effects of Some Medicinal Plants: Anacardium occidentale, Eucalyptus globulus, Psidium guajava, and Xylopia aethiopica Extracts in Alloxan-Induced Diabetic Male Wistar Albino Rats Thu, 27 Nov 2014 00:10:04 +0000 Insulin therapy and oral antidiabetic agents/drugs used in the treatment of diabetes mellitus have not sufficiently proven to control hyperlipidemia, which is commonly associated with the diabetes mellitus. Again the hopes that traditional medicine and natural plants seem to trigger researchers in this area is yet to be discovered. This research was designed to compare the biochemical effects of some medicinal plants in alloxan-induced diabetic male Wistar rats using named plants that are best at lowering blood glucose and hyperlipidemia and ameliorating other complications of diabetes mellitus by methods of combined therapy. The results obtained showed 82% decrease in blood glucose concentration after the 10th hour to the fortieth hour. There was significant increase in the superoxide dismutase activity of the test group administered 100 mg/kg of A. Occidentale. There was no significant difference recorded in the glutathione peroxidase activity of E. globulus (100 mg/kg) when compared to the test groups of P. guajava (250 mg/kg) and X. aethiopica (250 mg/kg). Catalase activity showed significant increase in the catalase activity, compared to test groups. While at , there was no significant difference seen between test group and treated groups. Meanwhile, degree of significance was observed in other parameters analysed. The biochemical analysis conducted in this study showed positive result, attesting to facts from previous works. Though these individual plants extracts exhibited significant increase in amelorating diabetes complication and blood glucose control compared to glibenclamide, a synthetic antidiabetic drug. Greater performance was observed in the synergy groups. Therefore, a poly/combined formulation of these plants extracts yielded significant result as well as resolving some other complications associated with diabetics. Victor Eshu Okpashi, Bayim Peter-Robins Bayim, and Margaret Obi-Abang Copyright © 2014 Victor Eshu Okpashi et al. All rights reserved. Evaluating the Biodeterioration Enzymatic Activities of Fungal Contamination Isolated from Some Ancient Yemeni Mummies Preserved in the National Museum Thu, 13 Nov 2014 06:44:40 +0000 Sophisticated mummification using chemical preservation was prevalent in ancient Yemeni civilization as noted in the 4th century B.C. mummies of the National Museum of Yemen, Sana’a, used in this study. Five of these mummies were used to evaluate hydrolytic enzymes produced as a result of fungal contamination. Forty-seven fungal species were isolated, thereby reflecting a high degree of contamination which may have resulted from the poor ventilation and preservation system. Aspergillus was the most common genus isolated (48.9%). Fifteen isolates exhibited ability to produce cellulase (EC;, Aspergillus candidus being the highest cellulose-producer. Pectin lyase (PL, EC; and pectin methyl esterase (PME, EC; were produced by Trichoderma hamatum, whereas chitinase (EC; was produced by Aspergillus niger. Protease activity was noted by only Cladosporium herbarum. The higher activities of these fungal hydrolytic enzymes represent the major threats of biodeterioration including deteriorating linen bandages as well as the mummy bodies. Therefore, it is recommended to improve the preservation system of the mummies at the National Museum to minimize the contamination up to the lowest level and protect the mummies from biodeterioration. Khalid Mohammed Naji, Qais Yusuf M. Abdullah, Aida Qaseem M. AL-Zaqri, and Saeed M. Alghalibi Copyright © 2014 Khalid Mohammed Naji et al. All rights reserved. Improved Rifamycin B Production by Nocardia mediterranei MTCC 14 under Solid-State Fermentation through Process Optimization Thu, 09 Oct 2014 14:31:12 +0000 Optimization of various production parameters using response surface methodology (RSM) was performed to assess maximum yield of rifamycin B from Nocardia mediterranei MTCC 14. Plackett-Burman design test was applied to determine the significant effects of various production parameters such as glucose, maltose, ribose, galactose, beef extract, peanut meal, ammonium chloride, ammonium sulphate, barbital, pH, and moisture content on production of rifamycin B. Among the eleven variables tested, galactose, ribose, glucose, and pH were found to have significant effect on rifamycin B production. Optimum levels of the significant variables were decided by using a central composite design. The most appropriate condition for production of rifamycin B was found to be a single step production at galactose (8% w/w), ribose (3% w/w), glucose (9% w/w), and pH (7.0). At these optimum production parameters, the maximum yield of rifamycin B obtained experimentally (9.87 g/kgds dry sunflower oil cake) was found to be very close to its predicted value of 10.35 g/kgds dry sunflower oil cake. The mathematical model developed was found to fit greatly with the experimental data of rifamycin B production. Basavaraj M. Vastrad, Shivayogeshwar E. Neelagund, Sudhir R. Iiger, Ajeet M. Godbole, and Venkatrao Kulkarni Copyright © 2014 Basavaraj M. Vastrad et al. All rights reserved. Serum Prolidase Activity and Oxidative Stress in Diabetic Nephropathy and End Stage Renal Disease: A Correlative Study with Glucose and Creatinine Mon, 08 Sep 2014 05:46:27 +0000 Association of oxidative stress and serum prolidase activity (SPA) has been reported in many chronic diseases. The study was aimed at evaluating the correlation of glucose and creatinine to SPA and oxidative stress in patients with diabetic nephropathy (DN) and end stage renal disease (ESRD) concerned with T2DM. 50 healthy volunteers, 50 patients with T2DM, 86 patients with DN, and 43 patients with ESRD were considered as control-1, control-2, case-1, and case-2, respectively. Blood glucose, creatinine, SPA, total oxidant status (TOS), total antioxidant status (TAS), and oxidative stress index (OSI) were measured by colorimetric tests. SPA, TOS, and OSI were significantly increased in case-1 and case-2 than control-1 and control-2, while TAS was significantly decreased . Blood glucose was linearly correlated to SPA, TOS, TAS, and OSI in control-2, case-1 and case-2 . Serum creatinine was linearly correlated with SPA, TOS, TAS and OSI in control-2 and case-1 . In case-2, serum creatinine was significantly correlated with SPA only . Thus, the study concluded that SPA and oxidative stress significantly correlated with blood glucose and creatinine. SPA, TOS, TAS, and OSI can be used as biomarkers for diagnosis of kidney damage. Akhilesh Kumar Verma, Subhash Chandra, Rana Gopal Singh, Tej Bali Singh, Shalabh Srivastava, and Ragini Srivastava Copyright © 2014 Akhilesh Kumar Verma et al. All rights reserved. A Polymorphism in Hepatocyte Nuclear Factor 1 Alpha, rs7310409, Is Associated with Left Main Coronary Artery Disease Mon, 18 Aug 2014 06:20:48 +0000 Coronary artery disease is the leading cause of mortality and morbidity in the world. Left main coronary artery disease (LMCAD) is a particularly severe phenotypic form of CAD and has a genetic basis. We hypothesized that some inflammation- and hyperhomocysteinemia-related gene polymorphisms may contribute to LMCAD susceptibility in a Chinese population. We studied the association between polymorphisms in the genes hepatocyte nuclear factor 1 alpha (HNF1A; rs7310409, G/A), C-reactive protein (rs1800947 and rs3093059 T/C), methylenetetrahydrofolate reductase (rs1801133, C/T), and methylenetetrahydrofolate dehydrogenase (rs1076991, A/G) in 402 LMCAD and 804 more peripheral CAD patients in a Chinese population. Genotyping was performed using the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry method. When the HNF1A rs7310409 GG homozygote genotype was used as the reference group, both the individual, GA and AA, and combined GA/AA genotypes were associated with an increased risk of LMCAD. This single nucleotide polymorphism (rs7310409) is strongly associated with plasma CRP levels. In conclusion, the present study provides evidence that the HNF1A rs7310409 G/A functional polymorphism may contribute to the risk of LMCAD. Rui Liu, Hanning Liu, Haiyong Gu, Xiao Teng, Yu Nie, Zhou Zhou, Yan Zhao, Shengshou Hu, and Zhe Zheng Copyright © 2014 Rui Liu et al. All rights reserved. A Metabolic Study on Colon Cancer Using 1H Nuclear Magnetic Resonance Spectroscopy Thu, 14 Aug 2014 11:18:23 +0000 Background. Colorectal carcinoma is the third cause of cancer deaths in the world. For diagnosis, invasive methods like colonoscopy and sigmoidoscopy are used, and noninvasive screening tests are not very accurate. We decided to study the potential of 1HNMR spectroscopy with metabolomics and chemometrics as a preliminary noninvasive test. We obtained a distinguishing pattern of metabolites and metabolic pathways between colon cancer patient and normal. Methods. Sera were obtained from confirmed colon cancer patients and the same number of healthy controls. Samples were sent for 1HNMR spectroscopy and analysis was carried out Chenomex and MATLAB software. Metabolites were identified using Human Metabolic Data Base (HDMB) and the main metabolic cycles were identified using Metaboanalyst software. Results. 15 metabolites were identified such as pyridoxine, orotidine, and taurocholic acid. Main metabolic cycles involved were the bile acid biosynthesis, vitamin B6 metabolism, methane metabolism, and glutathione metabolism. Discussion. The main detected metabolic cycles were also reported earlier in different cancers. Our observations corroborated earlier studies that suggest the importance of lowering serum LCA/DCA and increasing vitamin B6 intake to help prevent colon cancer. This work can be looked upon as a preliminary step in using 1HNMR analysis as a screening test before invasive procedures. Zahra Zamani, Mohammad Arjmand, Farideh Vahabi, Seyed Mahmood Eshaq Hosseini, Sadegh Mohammad Fazeli, Ayda Iravani, Parastoo Bayat, Akbar Oghalayee, Mahshid Mehrabanfar, Reza Haj Hosseini, Mohammad Tashakorpour, Mohsen Tafazzoli, and Sedigheh Sadeghi Copyright © 2014 Zahra Zamani et al. All rights reserved. Green Tea Potentially Ameliorates Bisphenol A-Induced Oxidative Stress: An In Vitro and In Silico Study Sun, 10 Aug 2014 11:12:27 +0000 The present investigation was an attempt to elucidate oxidative stress induced by bisphenol A on erythrocytes and its amelioration by green tea extract. For this, venous blood samples from healthy human adults were collected in EDTA vials and used for preparation of erythrocytes suspension. When erythrocyte suspensions were treated with different concentrations of BPA/, a dose-dependent increase in hemolysis occurred. Similarly, when erythrocytes suspensions were treated with either different concentrations of (0.05–0.25 mM) along with BPA (50 μg/mL) or 0.05 mM along with different concentrations of BPA (50–250 μg/mL), dose-dependent significant increase in hemolysis occurred. The effect of BPA and was found to be additive. For the confirmation, binding capacity of bisphenol A with erythrocyte proteins (hemoglobin, catalase, and glutathione peroxidase) was inspected using molecular docking tool, which showed presence of various hydrogen bonds of BPA with the proteins. The present data clearly indicates that BPA causes oxidative stress in a similar way as . Concurrent addition of different concentrations (10–50 μg/mL) of green tea extract to reaction mixture containing high dose of bisphenol A (250 μg/mL) caused concentration-dependent amelioration in bisphenol A-induced hemolysis. The effect was significant (). It is concluded that BPA-induced oxidative stress could be significantly mitigated by green tea extract. Hiral Suthar, R. J. Verma, Saumya Patel, and Y. T. Jasrai Copyright © 2014 Hiral Suthar et al. All rights reserved. Cytotoxic Effects of Newly Synthesized Palladium(II) Complexes of Diethyldithiocarbamate on Gastrointestinal Cancer Cell Lines Thu, 24 Jul 2014 10:12:01 +0000 As a part of a drug development program to discover novel therapeutic and more effective palladium (Pd) based anticancer drugs, a series of water-soluble Pd complexes have been synthesized by interaction between [Pd (phen)(H2O)2(NO3)2] and alkylenebisdithiocarbamate(al-bis-dtc) disodium salts. This study was undertaken to examine the possible cytotoxic effect of three novel complexes (0.125–64 µg/mL) on human gastric carcinoma (AGS), esophageal squamous cell carcinoma (Kyse-30), and hepatocellular carcinoma (HepG2) cell lines. The cytotoxicity was examined using cell proliferation and acridine orange/ethidium bromide (AO/EB) assay. In order to examine the effects of new Pd(II) complexes on cell cycle status, we performed cell cycle analysis. The complexes were found to have completely lethal effects on the cell lines, and the half maximal inhibitory concentration (IC50) values obtained for the cell lines were much lower in comparison with cisplatin. We demonstrated that the three new Pd(II) complexes are able to induce G2/M phase arrest in AGS and HepG2; in addition, the Pd(II) complexes caused an S phase arrest in Kyse-30 cell line. Our results indicate that newly synthesized Pd(II) complexes may provide a novel class of chemopreventive compounds for anticancer therapy. Shahram Hadizadeh, Nowruz Najafzadeh, Mohammad Mazani, Mojtaba Amani, Hassan Mansouri-Torshizi, and Ali Niapour Copyright © 2014 Shahram Hadizadeh et al. All rights reserved. Influence of Polymorphism on Glycosylation of Serum Amyloid A4 Protein Thu, 15 May 2014 11:35:56 +0000 Serum amyloid A4 (SAA4) is a constitutive apolipoprotein of high-density lipoprotein. It exhibits N-linked glycosylation in its second half. There are both glycosylated and nonglycosylated forms in plasma and the ratio of these two forms varies among individuals. This study was conducted to examine the influence of genetic polymorphism of SAA4 on its glycosylation status. In 55 healthy subjects, SAA4 polymorphism was analyzed by PCR combined direct sequencing and its glycosylation status was analyzed by immunoblotting. The results showed that the percentage of glycosylation in subjects with amino acid substitutions at positions 71 and/or 84 was significantly () higher than that in subjects with the wild type. The polymorphism had no influence on the plasma concentration of SAA4. These findings suggest that the changes in protein structures alter the efficiency of glycosylation in the SAA4 molecule. The functional implication of this should be of interest. Toshiyuki Yamada, Jyunji Sato, Kazuhiko Kotani, and Masafumi Tanaka Copyright © 2014 Toshiyuki Yamada et al. All rights reserved. Identification of Potential Herbal Inhibitor of Acetylcholinesterase Associated Alzheimer’s Disorders Using Molecular Docking and Molecular Dynamics Simulation Wed, 14 May 2014 06:48:39 +0000 Cholinesterase inhibitors (ChE-Is) are the standard for the therapy of AD associated disorders and are the only class of approved drugs by the Food and Drug Administration (FDA). Additionally, acetylcholinesterase (AChE) is the target for many Alzheimer’s dementia drugs which block the function of AChE but have some side effects. Therefore, in this paper, an attempt was made to elucidate cholinesterase inhibition potential of secondary metabolite from Cannabis plant which has negligible or no side effect. Molecular docking of 500 herbal compounds, against AChE, was performed using Autodock 4.2 as per the standard protocols. Molecular dynamics simulations have also been carried out to check stability of binding complex in water for 1000 ps. Our molecular docking and simulation have predicted high binding affinity of secondary metabolite () to AChE. Further, molecular dynamics simulations for 1000 ps suggest that ligand interaction with the residues Asp72, Tyr70-121-334, and Phe288 of AChE, all of which fall under active site/subsite or binding pocket, might be critical for the inhibitory activity of AChE. This approach might be helpful to understand the selectivity of the given drug molecule in the treatment of Alzheimer's disease. The study provides evidence for consideration of as a valuable small ligand molecule in treatment and prevention of AD associated disorders and further in vitro and in vivo investigations may prove its therapeutic potential. Chandrabhan Seniya, Ghulam Jilani Khan, and Kuldeep Uchadia Copyright © 2014 Chandrabhan Seniya et al. All rights reserved. Microbial Tyrosinases: Promising Enzymes for Pharmaceutical, Food Bioprocessing, and Environmental Industry Tue, 06 May 2014 06:57:35 +0000 Tyrosinase is a natural enzyme and is often purified to only a low degree and it is involved in a variety of functions which mainly catalyse the o-hydroxylation of monophenols into their corresponding o-diphenols and the oxidation of o-diphenols to o-quinones using molecular oxygen, which then polymerizes to form brown or black pigments. The synthesis of o-diphenols is a potentially valuable catalytic ability and thus tyrosinase has attracted a lot of attention with respect to industrial applications. In environmental technology it is used for the detoxification of phenol-containing wastewaters and contaminated soils, as biosensors for phenol monitoring, and for the production of L-DOPA in pharmaceutical industries, and is also used in cosmetic and food industries as important catalytic enzyme. Melanin pigment synthesized by tyrosinase has found applications for protection against radiation cation exchangers, drug carriers, antioxidants, antiviral agents, or immunogen. The recombinant V. spinosum tryosinase protein can be used to produce tailor-made melanin and other polyphenolic materials using various phenols and catechols as starting materials. This review compiles the recent data on biochemical and molecular properties of microbial tyrosinases, underlining their importance in the industrial use of these enzymes. After that, their most promising applications in pharmaceutical, food processing, and environmental fields are presented. Kamal Uddin Zaidi, Ayesha S. Ali, Sharique A. Ali, and Ishrat Naaz Copyright © 2014 Kamal Uddin Zaidi et al. All rights reserved. Exploring Drug Targets in Isoprenoid Biosynthetic Pathway for Plasmodium falciparum Wed, 23 Apr 2014 11:46:54 +0000 Emergence of rapid drug resistance to existing antimalarial drugs in Plasmodium falciparum has created the need for prediction of novel targets as well as leads derived from original molecules with improved activity against a validated drug target. The malaria parasite has a plant plastid-like apicoplast. To overcome the problem of falciparum malaria, the metabolic pathways in parasite apicoplast have been used as antimalarial drug targets. Among several pathways in apicoplast, isoprenoid biosynthesis is one of the important pathways for parasite as its multiplication in human erythrocytes requires isoprenoids. Therefore targeting this pathway and exploring leads with improved activity is a highly attractive approach. This report has explored progress towards the study of proteins and inhibitors of isoprenoid biosynthesis pathway. For more comprehensive analysis, antimalarial drug-protein interaction has been covered. Tabish Qidwai, Farrukh Jamal, Mohd Y. Khan, and Bechan Sharma Copyright © 2014 Tabish Qidwai et al. All rights reserved. A Nonhost Peptidase Inhibitor of ~14 kDa from Butea monosperma (Lam.) Taub. Seeds Affects Negatively the Growth and Developmental Physiology of Helicoverpa armigera Thu, 17 Apr 2014 13:01:37 +0000 Helicoverpa armigera is one of the major devastating pests of crop plants. In this context a serine peptidase inhibitor purified from the seeds of Butea monosperma was evaluated for its effect on developmental physiology of H. armigera larvae. B. monosperma peptidase inhibitor on 12% denaturing polyacrylamide gel electrophoresis exhibited a single protein band of ~14 kDa with or without reduction. In vitro studies towards total gut proteolytic enzymes of H. armigera and bovine trypsin indicated measurable inhibitory activity. B. monosperma peptidase inhibitor dose for 50% mortality and weight reduction by 50% were 0.5% w/w and 0.10% w/w, respectively. The IC50 of B. monosperma peptidase inhibitor against total H. armigera gut proteinases activity was 2.0 µg/mL. The larval feeding assays suggested B. monosperma peptidase inhibitor to be toxic as reflected by its retarded growth and development, consequently affecting fertility and fecundity of pest and prolonging the larval-pupal duration of the insect life cycle of H. armigera. Supplementing B. monosperma peptidase inhibitor in artificial diet at 0.1% w/w, both the efficiencies of conversion of ingested as well as digested food were downregulated, whereas approximate digestibility and metabolic cost were enhanced. The efficacy of Butea monosperma peptidase inhibitor against progressive growth and development of H. armigera suggest its usefulness in insect pest management of food crops. Prabhash K. Pandey, Dushyant Singh, Sangram Singh, M. Y. Khan, and Farrukh Jamal Copyright © 2014 Prabhash K. Pandey et al. All rights reserved. Roles of the Oxidative Stress and ADMA in the Development of Deep Venous Thrombosis Sun, 13 Apr 2014 00:00:00 +0000 Venous thromboembolism has multifactorial origin and occurs in the context of complex interactions between environmental and genetic predisposing factors. Oxidative stress plays an important role in the physiopathology of venous thrombosis. Current study examined the role of oxidative stress and asymmetric dimethylarginine in the development of DVT with the parameters such as serum malondialdehyde (MDA), glutathione peroxidase (GSH-Px), catalase, ADMA, homocysteine, folic acid, vitamin B6, and vitamin B12 levels. Serum MDA levels were found significantly () high in patients with DVT compared with control group. Additionally, serum B6 levels were found significantly () low in patients with DVT compared with healthy volunteers. There were no significant differences between the groups in terms of the other parameters (). This study showed that patients with DVT have increased oxidative stress compared with the healthy volunteers whereas there was no significant difference between the groups in terms of serum ADMA levels. Thus serum ADMA levels seemed to be not related with development of DVT. Meral Ekim, M. Ramazan Sekeroglu, Ragıp Balahoroglu, Halil Ozkol, and Hasan Ekim Copyright © 2014 Meral Ekim et al. All rights reserved. Identification and Characterization of TEX101 in Bovine Epididymal Spermatozoa Thu, 10 Apr 2014 17:42:22 +0000 Several studies exhibit the presence of Ricinus Communis Agglutinin I (RCA) binding glycocalyx in mammalian spermatozoa. However, the molecular characterization of RCA binding glycocalyx in sperm membranes and its mechanism of action are poorly understood. The objective of the study was to identify and to characterize RCA binding glycoprotein of the bovine sperm plasma membranes (PM). Lectin blots of caput and cauda sperm PM revealed a 38 kDa polypeptide exhibiting the highest affinity to RCA among the several major RCA binding polypeptides. The 38 kDa RCA binding polypeptide of cauda sperm PM was purified and exhibited a charge train of three distinct spots with isoelectric points (pH 5.3 and 5.8). Proteomic identification yielded ten peptides that matched the sequence of Testis Expressed 101 protein (TEX101). Western blots data revealed that bovine sperm TEX101 is present in both testicular and epididymal sperm PM fractions. The native TEX101 polypeptide contains ~17 kDa N-linked oligosaccharides and the polypeptide is anchored to sperm membrane via a glycosylphosphatidylinositol lipid linkage. Immunofluorescence staining of sperm with anti-TEX101 demonstrated that the polypeptide is localized at the head of cauda sperm. Our biochemical results provide evidence on the presence of TEX101 in bovine epididymal sperm plasma membranes and may have a potential role in sperm-egg interaction. Subir K. Nagdas, Eric L. McLean, Leeá P. Richardson, and Samir Raychoudhury Copyright © 2014 Subir K. Nagdas et al. All rights reserved. A Comparative Study of Phase States of the Peribacteroid Membrane from Yellow Lupin and Broad Bean Nodules Thu, 03 Apr 2014 08:57:43 +0000 A comparative study of the lipid bilayer phase status and structure of the outer membrane of free-living Bradyrhizobium strain 359a (Nod+Fix+) and 400 (Nod+FixL) or Rhizobium leguminosarum 97 (Nod+Fix+, effective) and 87 (Nod+FixL, ineffective) has been carried out. Also, the effect of the symbiotic pair combination on the lipid bilayer structure of the bacteroid outer membrane and peribacteroid membrane, isolated from the nodules of Lupinus luteus L. or Vicia faba L., has been studied. As a result, it is shown that the lipid bilayer status of the bacteroid outer membrane is mainly determined by microsymbiont, but not the host plant. In the contrast, the lipid bilayer status of the peribacteroid membrane and, as a consequence, its properties depend on interaction of both symbiotic partners. Natalia N. Kudryavtseva, Alexis V. Sof’in, Georgiy S. Bobylev, and Evgeny M. Sorokin Copyright © 2014 Natalia N. Kudryavtseva et al. All rights reserved. Adaptation of Sonication-Assisted Matrix Solid Phase Dispersion of Tissues for the Subsequent Extraction of Polycyclic Aromatic Hydrocarbons from Gulf Menhaden (Brevoortia patronus) Sun, 09 Mar 2014 10:13:48 +0000 A new adaptation based on matrix solid phase dispersion of tissue for the subsequent isolation of polycyclic aromatic hydrocarbons was developed and used for extractions of Gulf menhaden caught during the summer of 2011. Many Matrix Solid Phase Dispersion (MSPD) methods require specific cartridges and other clean-up materials in order to achieve proper extraction. For this study, the tissues were lyophilized prior to applying the adapted MSPD method allowing for a much more complete homogenization with the C18 silica. The tissue was spiked with phenanthrene d10 as a surrogate as a measure of PAH recovery prior to the lyophilisation process to determine if any target compounds were lost and prior to sonication as per the finalized adaptation procedure to determine method efficiency. This technique used C18 silica in a 1 : 1 ratio as the primary homogenizing material for the menhaden tissue matrix and was eluted with dichloromethane (DCM) until visibly clear. The overall study mean recovery was with method detection limits between 0.4 ng/g and 4.4 ng/g tissue dry weight. This adapted protocol has been used exclusively on the analysis of high lipid content fish stocks affected by dispersed and weathered oil from the BP Horizon incident. Gregory M. Olson, Buffy M. Meyer, and Ralph J. Portier Copyright © 2014 Gregory M. Olson et al. All rights reserved. Modeling Dendrimers Charge Interaction in Solution: Relevance in Biosystems Thu, 27 Feb 2014 07:00:20 +0000 Dendrimers are highly branched macromolecules obtained by stepwise controlled, reaction sequences. The ability to be designed for specific applications makes dendrimers unprecedented components to control the structural organization of matter during the bottom-up synthesis of functional nanostructures. For their applications in the field of biotechnology the determination of dendrimer structural properties as well as the investigation of the specific interaction with guest components are needed. We show how the analysis of the scattering structure factor S(q), in the framework of current models for charged systems in solution, allows for obtaining important information of the interdendrimers electrostatic interaction potential. The finding of the presented results outlines the important role of the dendrimer charge and the solvent conditions in regulating, through the modulation of the electrostatic interaction potential, great part of the main structural properties. This charge interaction has been indicated by many studies as a crucial factor for a wide range of structural processes involving their biomedical application. Due to their easily controllable properties dendrimers can be considered at the crossroad between traditional colloids, associating polymers, and biological systems and represent then an interesting new technological approach and a suitable model system of molecular organization in biochemistry and related fields. Domenico Lombardo Copyright © 2014 Domenico Lombardo. All rights reserved. Expression of uPA, tPA, and PAI-1 in Calcified Aortic Valves Mon, 17 Feb 2014 06:52:35 +0000 Purpose. Our physiopathological assumption is that u-PA, t-PA, and PAI-1 are released by calcified aortic valves and play a role in the calcification of these valves. Methods. Sixty-five calcified aortic valves were collected from patients suffering from aortic stenosis. Each valve was incubated for 24 hours in culture medium. The supernatants were used to measure u-PA, t-PA, and PAI-1 concentrations; the valve calcification was evaluated using biphotonic absorptiometry. Results. Aortic stenosis valves expressed normal plasminogen activators concentrations and overexpressed PAI-1 (u-PA, t-PA, and PAI-1 mean concentrations were, resp., 1.69 ng/mL ± 0.80, 2.76 ng/mL ± 1.33, and 53.27 ng/mL ± 36.39). There was no correlation between u-PA and PAI-1 () but t-PA and PAI-1 were strongly correlated with each other (). Overexpression of PAI-1 was proportional to the calcium content of the AS valves. Conclusions. Our results demonstrate a consistent increase of PAI-1 proportional to the calcification. The overexpression of PAI-1 may be useful as a predictive indicator in patients with aortic stenosis. Najlah Kochtebane, Abdullah Mossa M. Alzahrani, and Aghleb Bartegi Copyright © 2014 Najlah Kochtebane et al. All rights reserved. Combination Therapies for the Treatment of Advanced Melanoma: A Review of Current Evidence Wed, 12 Feb 2014 10:07:43 +0000 The treatment of advanced melanoma has been revolutionised in recent years with the advent of a range of new therapies. BRAF inhibitors, such as vemurafenib, have demonstrated improvements in the overall survival of patients with advanced melanoma that harbour a BRAF V600 mutation. Alongside these targeted therapies, novel immune-checkpoint inhibitors, such as ipilimumab, have also been developed and have produced similarly improved outcomes for patients. For the first time in the history of melanoma, monotherapy with each of these drugs has produced improvements in the overall survival of patients with advanced disease. Building on this initial success, there has been intense interest in developing combination therapies predominantly with either dual blockade of the MAPK oncogenic pathway or dual immune-checkpoint blockade. The current evidence for the use of these combination therapies will be presented here. Mark Voskoboynik and Hendrik-Tobias Arkenau Copyright © 2014 Mark Voskoboynik and Hendrik-Tobias Arkenau. All rights reserved. Production and Partial Purification of Alpha Amylase from Bacillus subtilis (MTCC 121) Using Solid State Fermentation Sun, 12 Jan 2014 12:48:47 +0000 Amylase is an enzyme that catalyzes the breakdown of starch into sugars and plays a pivotal role in a variety of areas like use as digestives, for the production of ethanol and high fructose corn syrup, detergents, desiring of textiles, modified starches, hydrolysis of oil-field drilling fluids, and paper recycling. In the present work, solid state fermentation (SSF) for α-amylase production has been used in lieu of submerged fermentation (SmF) due to its simple technique, low capital investment, lower levels of catabolite repression, and better product recovery. Bacillus subtilis has been well known as producer of alpha amylase and was tested using solid state fermentation for 48 hours at 37°C with wheat bran as substrate. Comparison between different fermentation hours demonstrated high yield of alpha amylase after 48 hours. This alpha amylase has optimum pH and temperature at 7.1 and 40°C, respectively. With the goal to purify alpha amylase, 30–70% (NH4)2SO4 cut concentrated the amylase activity threefold with respect to crude fermented extract. This was verified in quantitative DNS assay method as well as in zymogram gel profile. The exact molecular weight of the amylase is yet to be determined with the aid of other protein purification techniques. Dibyangana Raul, Tania Biswas, Suchita Mukhopadhyay, Shrayan Kumar Das, and Suvroma Gupta Copyright © 2014 Dibyangana Raul et al. All rights reserved.