Biochemistry Research International The latest articles from Hindawi Publishing Corporation © 2016 , Hindawi Publishing Corporation . 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. Differences in GPR30 Regulation by Chlorotriazine Herbicides in Human Breast Cells Wed, 03 Feb 2016 07:22:59 +0000 Over 200,000 cases of invasive breast cancer are diagnosed annually; herbicide contaminants in local water sources may contribute to the growth of these cancers. GPR30, a G protein coupled receptor, was identified as a potential orphan receptor that may interact with triazine herbicides such as atrazine, one of the most commonly utilized chlorotriazines in agricultural practices in the United States. Our goal was to identify whether chlorotriazines affected the expression of GPR30. Two breast cancer cell lines, MDA-MB-231 and MCF-7, as well as one normal breast cell line, MCF-10A, were treated with a 100-fold range of atrazine, cyanazine, or simazine, with levels flanking the EPA safe level for each compound. Using real-time PCR, we assessed changes in GPR30 mRNA compared to a GAPDH control. Our results indicate that GPR30 expression increased in breast cancer cells at levels lower than the US EPA drinking water contamination limit. During this treatment, the viability of cells was unaltered. In contrast, treatment with chlorotriazines reduced the expression of GPR30 in noncancerous MCF-10A cells. Thus, our results indicate that cell milieu and potential to metastasize may play a role in the extent of GPR30 response to pesticide exposure. Colin P. Florian, Shelly R. Mansfield, and Jennifer R. Schroeder Copyright © 2016 Colin P. Florian et al. All rights reserved. Lipidomic Modulation in Stressed Albino Rats Is Altered by Yolk and Albumen of Quail (Coturnix japonica) Egg and Poultry Feed Thu, 28 Jan 2016 13:27:23 +0000 Cold and immobilization stressors can generate oxidative stress as well as skeletal muscle fatigue. Free radicals cause oxidative degradation of lipids, proteins, nucleic acids, and carbohydrates molecules, thereby compromising cell integrity and function. Quail egg had been described as being very functional biochemically, due to the essential biomolecules it contains in very regulated quantity. This study was aimed for evaluating the dietary effect of the egg on lipid profile parameters on selected tissues. The antilipidemic properties of the egg yolk and albumen and poultry (layers) feed were determined in selected tissues in male albino rats assaulted with cold immobilization stress induced on them at 4°C for 2 hours, while diazepam was used as standard antistress drug. Antilipidemic activities were evaluated by lipid profile modulation (HDL, LDL, TRIG., and T-CHOL.). Quantitative and qualitative analyses of fatty acids profile of the yolk hexane-extract were determined by Gas Chromatography and Mass Spectrophotometry (GC-MS). The ameliorative impacts of diazepam (2.5 and 5.0 mg/mL/kg BW), yolk (5 and 10 mL/kg BW), albumen (5 and 10 mL/kg BW), and the feed (5–10 mg/kg BW) were competitively () specific for each of the tissues. The result of the study suggested yolk and albumen of quail egg and poultry feed as antistress agents as well as lipid modulators. Emmanuel Oluwafemi Ibukun and Gideon Oludare Oladipo Copyright © 2016 Emmanuel Oluwafemi Ibukun and Gideon Oludare Oladipo. All rights reserved. Inactivation of Anopheles gambiae Glutathione Transferase ε2 by Epiphyllocoumarin Tue, 26 Jan 2016 16:15:34 +0000 Glutathione transferases (GSTs) are part of a major family of detoxifying enzymes that can catalyze the reductive dehydrochlorination of dichlorodiphenyltrichloroethane (DDT). The delta and epsilon classes of insect GSTs have been implicated in conferring resistance to this insecticide. In this study, the inactivation of Anopheles gambiae GSTε2 by epiphyllocoumarin (Tral 1) was investigated. Recombinant AgGSTε2 was expressed in Escherichia coli cells containing a pET3a-AGSTε2 plasmid and purified by affinity chromatography. Tral 1 was shown to inactivate GSTε2 both in a time-dependent manner and in a concentration-dependent manner. The half-life of GSTε2 in the presence of 25 μM ethacrynic acid (ETA) was 22 minutes and with Tral 1 was 30 minutes, indicating that Tral 1 was not as efficient as ETA as an inactivator. The inactivation parameters and were found to be 0.020 ± 0.001 min−1 and 7.5 ± 2.1 μM, respectively, after 90 minutes of incubation. Inactivation of GSTε2 by Tral 1 implies that Tral 1 covalently binds to this enzyme in vitro and would be expected to exhibit time-dependent effects on the enzyme in vivo. Tral 1, therefore, would produce irreversible effects when used together with dichlorodiphenyltrichloroethane (DDT) in malaria control programmes where resistance is mediated by GSTs. Patience Marimo, Rose Hayeshi, and Stanley Mukanganyama Copyright © 2016 Patience Marimo et al. All rights reserved. Novel 4-Thiazolidinone Derivatives as Anti-Infective Agents: Synthesis, Characterization, and Antimicrobial Evaluation Tue, 26 Jan 2016 09:52:02 +0000 A series of new 4-thiazolidinone derivatives was synthesized, characterized by spectral techniques, and screened for antimicrobial activity. All the compounds were evaluated against five Gram-positive bacteria, two Gram-negative bacteria, and two fungi, at concentrations of 50, 100, 200, 400, 800, and 1600 µg/mL, respectively. Minimum inhibitory concentrations of all the compounds were also determined and were found to be in the range of 100–400 µg/mL. All the compounds showed moderate-to-good antimicrobial activity. Compounds 4a [2-(4-fluoro-phenyl)-3-(4-methyl-5,6,7,8-tetrahydro-quinazolin-2-yl)-thiazolidin-4-one] and 4e [3-(4,6-dimethyl-pyrimidin-2-yl)-2-(2-methoxy-phenyl)-thiazolidin-4-one] were the most potent compounds of the series, exhibiting marked antimicrobial activity against Pseudomonas fluorescens, Staphylococcus aureus, and the fungal strains. Thus, on the basis of results obtained, it may be concluded that synthesized compounds exhibit a broad spectrum of antimicrobial activity. Amit Gupta, Rajendra Singh, Pankaj K. Sonar, and Shailendra K. Saraf Copyright © 2016 Amit Gupta et al. All rights reserved. Evaluation of Early Markers of Nephropathy in Patients with Type 2 Diabetes Mellitus Sun, 24 Jan 2016 12:31:38 +0000 Aims. T2DM often remains undiagnosed for many years because hyperglycemia develops gradually and may not produce any symptoms. As patients with T2DM are at increased risk of microvascular and macrovascular complications, the preclinical diagnosis of the state is the key point of the disease management. Methods. We evaluated parameters such as GAGs/PGs, NAG, and NGAL in urine samples from 43 normoalbuminuric T2DM patients and 31 apparently healthy control subjects. Results. The total urinary GAG excretion showed no significant differences between patients and controls. The electrophoretic analysis evidenced the presence of UTI and its degradation products (LSC and SM-LSC), CS, and HS. We observed modifications of HS and total UTI (including UTI and its degradation products) relative contents in T2DM patients compared with controls whereas no differences in CS percentage were found. NGAL levels were significantly increased in T2DM patients and were positively correlated with both NAG (, ) and the presence of hypertension (, ). Conclusions. These data suggest that the assessed molecules could represent useful markers to detect early renal impairment in patients with T2DM. Pierina De Muro, Antonio Junior Lepedda, Gabriele Nieddu, Michela Idini, Hai Quy Tram Nguyen, Omar Lobina, Pietro Fresu, and Marilena Formato Copyright © 2016 Pierina De Muro et al. All rights reserved. Modulation of Erythrocyte Plasma Membrane Redox System Activity by Curcumin Tue, 19 Jan 2016 18:24:38 +0000 Plasma membrane redox system (PMRS) is an electron transport chain system ubiquitously present throughout all cell types. It transfers electron from intracellular substrates to extracellular acceptors for regulation of redox status. Curcumin, isolated from Curcuma longa, has modulatory effects on cellular physiology due to its membrane interaction ability and antioxidant potential. The present study investigates the effect of curcumin on PMRS activity of erythrocytes isolated from Wistar rats in vitro and in vivo and validated through an in silico docking simulation study using Molegro Virtual Docker (MVD). Effects of curcumin were also evaluated on level of glutathione (GSH) and the oxidant potential of plasma measured in terms of plasma ferric equivalent oxidative potentials (PFEOP). Results show that curcumin significantly () downregulated the PMRS activity in a dose-dependent manner. Molecular docking results suggest that curcumin interacts with amino acids at the active site cavity of cytochrome reductase, a key constituent of PMRS. Curcumin also increased the GSH level in erythrocytes and plasma while simultaneously decreasing the oxidant potential (PFEOP) of plasma. Altered PMRS activity and redox status are associated with the pathophysiology of several health complications including aging and diabetes; hence, the above finding may explain part of the role of curcumin in health beneficial effects. Prabhakar Singh, Rajesh Kumar Kesharwani, Krishna Misra, and Syed Ibrahim Rizvi Copyright © 2016 Prabhakar Singh et al. All rights reserved. Protective Effects of Carvacrol against Oxidative Stress Induced by Chronic Stress in Rat’s Brain, Liver, and Kidney Tue, 19 Jan 2016 16:20:11 +0000 Restraint stress may be associated with elevated free radicals, and thus, chronic exposure to oxidative stress may cause tissue damage. Several studies have reported that carvacrol (CAR) has a protective effect against oxidative stress. The present study was designed to investigate the protective effects of CAR on restraint stress induced oxidative stress damage in the brain, liver, and kidney. For chronic restraint stress, rats were kept in the restrainers for 6 h every day, for 21 consecutive days. The animals received systemic administrations of CAR daily for 21 days. To evaluate the changes of the oxidative stress parameters following restraint stress, the levels of malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and catalase (CAT) activities were measured in the brain, liver, and kidney. In the stressed animals that received vehicle, the MDA level was significantly higher () and the levels of GSH and antioxidant enzymes were significantly lower than the nonstressed animals (). CAR ameliorated the changes in the stressed animals as compared with the control group (). This study indicates that CAR can prevent restraint stress induced oxidative damage. Saeed Samarghandian, Tahereh Farkhondeh, Fariborz Samini, and Abasalt Borji Copyright © 2016 Saeed Samarghandian et al. All rights reserved. Fabrication and Optimization of ChE/ChO/HRP-AuNPs/c-MWCNTs Based Silver Electrode for Determining Total Cholesterol in Serum Mon, 18 Jan 2016 10:27:35 +0000 The developed method used three enzymes comprised of cholesterol esterase, cholesterol oxidase, and peroxidase for fabrication of amperometric biosensor in order to determine total cholesterol in serum samples. Gold nanoparticles (AuNPs) and carboxylated multiwall carbon nanotubes (cMWCNTs) were used to design core of working electrode, having covalently immobilized ChO, ChE, and HRP. Polyacrylamide layer was finally coated on working electrode in order to prevent enzyme leaching. Chemically synthesised Au nanoparticles were subjected to transmission electron microscopy (TEM) for analysing the shape and size of the particles. Working electrode was subjected to FTIR and XRD. The combined action of AuNP and c-MWCNT showed enhancement in electrocatalytic activity at a very low potential of 0.27 V. The pH 7, temperature 40°C, and response time of 20 seconds, respectively, were observed. The biosensor shows a broad linear range from 0.5 mg/dL to 250 mg/dL (0.01 mM–5.83 mM) with minimum detection limit being 0.5 mg/dL (0.01 mM). The biosensor showed reusability of more than 45 times and was stable for 60 days. The biosensor was successfully tested for determining total cholesterol in serum samples amperometrically with no significant interference by serum components. Kusum Lata, Vikas Dhull, and Vikas Hooda Copyright © 2016 Kusum Lata et al. All rights reserved. Halophilic Bacteria of Lunsu Produce an Array of Industrially Important Enzymes with Salt Tolerant Activity Mon, 18 Jan 2016 07:20:00 +0000 The halophilic bacterial isolates SS1, SS2, SS3, SS5, and SS8 were characterized for production of industrially important enzymes like amylase, protease, lipase, and glutaminase. Halophilic bacterial isolates SS1 and SS3 exhibited salt dependent extracellular amylase and protease activities. Both the halophilic isolates SS1 and SS3 exhibited maximum amylase and protease activities in the presence of 1.5 and 1.0 M NaCl, respectively, with the optimum pH 8 and temperature 40°C. SS2 showed maximum extracellular protease and lipase activities in the presence of 0.75 M NaCl, at optimum pH of 7, and temperature 37°C. The glutaminase activity of SS3 increased with increase in concentration of NaCl up to 2.5 M. The optimum pH and temperature for L-glutaminase activity of SS3 was 8 and 40°C, respectively. The combined hydrolytic activities of these halophilic bacterial isolates can be used for bioconversion of organic materials to useful products. Sonika Gupta, Parul Sharma, Kamal Dev, and Anuradha Sourirajan Copyright © 2016 Sonika Gupta et al. All rights reserved. Unravelling the Interactions between Hydrolytic and Oxidative Enzymes in Degradation of Lignocellulosic Biomass by Sporothrix carnis under Various Fermentation Conditions Mon, 11 Jan 2016 07:36:17 +0000 The mechanism underlying the action of lignocellulolytic enzymes in biodegradation of lignocellulosic biomass remains unclear; hence, it is crucial to investigate enzymatic interactions involved in the process. In this study, degradation of corn cob by Sporothrix carnis and involvement of lignocellulolytic enzymes in biodegradation were investigated over 240 h cultivation period. About 60% degradation of corn cob was achieved by S. carnis at the end of fermentation. The yields of hydrolytic enzymes, cellulase and xylanase, were higher than oxidative enzymes, laccase and peroxidase, over 144 h fermentation period. Maximum yields of cellulase (854.4 U/mg) and xylanase (789.6 U/mg) were at 96 and 144 h, respectively. Laccase and peroxidase were produced cooperatively with maximum yields of 489.06 U/mg and 585.39 U/mg at 144 h. Drastic decline in production of cellulase at 144 h (242.01 U/mg) and xylanase at 192 h (192.2 U/mg) indicates that they play initial roles in biodegradation of lignocellulosic biomass while laccase and peroxidase play later roles. Optimal degradation of corn cob (76.6%) and production of hydrolytic and oxidative enzymes were achieved with 2.5% inoculum at pH 6.0. Results suggest synergy in interactions between the hydrolytic and oxidative enzymes which can be optimized for improved biodegradation. Olusola A. Ogunyewo and Folasade M. Olajuyigbe Copyright © 2016 Olusola A. Ogunyewo and Folasade M. Olajuyigbe. 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