Biochemistry Research International The latest articles from Hindawi Publishing Corporation © 2014 , Hindawi Publishing Corporation . 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. Reference Ranges for Serum Uric Acid among Healthy Assamese People Thu, 09 Jan 2014 13:15:13 +0000 This study was designed to establish reference ranges for serum uric acid among healthy adult Assamese population. Samples from 1470 aged 35–86 years were used to establish age and sex related reference range by the centile method (central 95 percentile) for serum uric acid level. There were 51% () males and 49% () females; 75.9% () of them were from urban area and the rest 24.1% () were from the rural area. Majority of the population were nonvegetarian (98.6%, ) and only 1.4% () were vegetarian. The mean age, weight, height, and uric acid of the studied group were years,  kg,  cm, and  mg/dL, respectively. There is a statistically significant difference in the mean value of the abovementioned parameters between male and female. The observed reference range of uric acid in the population is 2.6–8.2 mg/dL which is wider than the current reference range used in the laboratory. Except gender (), we did not find any significant relation of uric acid with other selected factors. Madhumita Das, N. C. Borah, M. Ghose, and N. Choudhury Copyright © 2014 Madhumita Das et al. All rights reserved. Community Based Assessment of Biochemical Risk Factors for Cardiovascular Diseases in Rural and Tribal Area of Himalayan Region, India Sun, 22 Dec 2013 09:47:28 +0000 Context. Evident change in nutrition and lifestyle among individuals of urban and rural areas raises suspicion for similar change in tribal area population of India. Aim. To study the biochemical risk factor for CVDs in rural and tribal population of Sub-Himalayan state of India. Settings and Design. Cross-sectional study in rural (low altitude) and tribal (high altitude) area of Himachal Pradesh, India. Methodology. Blood lipid profile using standard laboratory methods. Statistical Analysis. Chi-square test and multiple linear regression analysis. Results. Total of 900 individuals were studied in both areas. As per Asian criteria, obesity (BMI 27.5–30.0 kg/m2) was observed to be significantly high () as 13.7% in tribal area as compared to 5.5% in rural area. Normal level of TC (<200 mg/dL) and LDL (<130 mg/dL) was observed in the majority of the population of both areas, whereas, at risk level of HDL (<40 mg/dL) was present in half of the population of both rural and tribal areas. The prevalence of borderline to high level of TGs was observed to be 60.2% and 55.2% in rural and tribal () area, respectively. Conclusion. Prevalent abnormal lipid profile in tribal area demands establishment of an effective surveillance system for development of chronic diseases. Ashok Kumar Bhardwaj, Dinesh Kumar, Sunil Kumar Raina, Pradeep Bansal, Satya Bhushan, and Vishav Chander Copyright © 2013 Ashok Kumar Bhardwaj et al. All rights reserved. Protective Effect of Theaflavin on Erythrocytes Subjected to In Vitro Oxidative Stress Sat, 21 Dec 2013 12:42:38 +0000 Antioxidant and free radical scavenging effect of black tea theaflavins has been shown in many epidemiological studies. In the present work we report the protective mechanism of tea theaflavins on biomarkers of oxidative stress, which are elevated during stress conditions. We hereby report the in vitro effect of theaflavins on erythrocyte malondialdehyde (MDA), intracellular reduced glutathione (GSH), and plasma membrane redox system (PMRS) of rats. The effect of theaflavin on PMRS has also been validated through an in silico docking simulation study using Molegro Virtual Docker (MVD). We report that theaflavins show significant protection to erythrocyte against oxidative stress induced by tert-butyl hydroperoxide (t-BHP). The findings suggest a possible protective role of theaflavins as antioxidant. Mahejabeen Fatima, Rajesh Kumar Kesharwani, Krishna Misra, and Syed Ibrahim Rizvi Copyright © 2013 Mahejabeen Fatima et al. All rights reserved. Jatropha Oil Derived Sophorolipids: Production and Characterization as Laundry Detergent Additive Sat, 21 Dec 2013 12:35:32 +0000 Sophorolipids (SLs) are glycolipidic biosurfactants suitable for various biological and physicochemical applications. The nonedible Jatropha oil has been checked as the alternative raw material for SL synthesis using C. bombicola (ATCC22214). This is useful towards lowering the SL production cost. Through optimization of fermentation parameters and use of resting cell method, the yield 15.25 g/L could be achieved for Jatropha oil derived SL (SLJO) with 1% v/v oil feeding. The synthesized SL displayed good surfactant property. It reduced the surface tension of distilled water from 70.7 mN/m to 33.5 mN/m with the Critical Micelle Concentration (CMC) value of 9.5 mg/L. Keeping the prospective use of the SL in mind, the physicochemical properties were checked along with emulsion stability under temperature, pH stress, and in hard water. Also antibacterial action and stain removal capability in comparison with commercial detergent was demonstrated. SLJO enhanced the detergent performance. Based on the results, it can be said that SLs have utility as fabric cleaner with advantageous properties such as skin friendly nature, antibacterial action, and biodegradability. Therefore SLs are potential green molecules to replace synthetic surfactants in detergents so as to reduce harm caused to environment through detergent usage. Kasturi Joshi-Navare, Poonam Khanvilkar, and Asmita Prabhune Copyright © 2013 Kasturi Joshi-Navare et al. All rights reserved. Penetration of Milk-Derived Antimicrobial Peptides into Phospholipid Monolayers as Model Biomembranes Tue, 17 Dec 2013 13:48:08 +0000 Three antimicrobial peptides derived from bovine milk proteins were examined with regard to penetration into insoluble monolayers formed with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) or 1,2-dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DPPG). Effects on surface pressure () and electric surface potential () were measured, with a platinum Wilhelmy plate and with a vibrating plate. The penetration measurements were performed under stationary diffusion conditions and upon the compression of the monolayers. The two type measurements showed greatly different effects of the peptide-lipid interactions. Results of the stationary penetration show that the peptide interactions with DPPC monolayer are weak, repulsive, and nonspecific while the interactions with DPPG monolayer are significant, attractive, and specific. These results are in accord with the fact that antimicrobial peptides disrupt bacteria membranes (negative) while no significant effect on the host membranes (neutral) is observed. No such discrimination was revealed from the compression isotherms. The latter indicate that squeezing the penetrant out of the monolayer upon compression does not allow for establishing the penetration equilibrium, so the monolayer remains supersaturated with the penetrant and shows an under-equilibrium orientation within the entire compression range, practically. Wanda Barzyk, Ewa Rogalska, and Katarzyna Więcław-Czapla Copyright © 2013 Wanda Barzyk et al. All rights reserved. Effect of Different Stages of Chronic Kidney Disease and Renal Replacement Therapies on Oxidant-Antioxidant Balance in Uremic Patients Thu, 12 Dec 2013 15:59:52 +0000 Oxidative stress seems to be involved in the path physiology of cardiovascular complications of chronic kidney disease (CKD). In this study, we determined the effect of different stages of CKD and substitutive therapies on oxidative stress. One hundred sixty-seven patients (age: years; male/female: 76/91) with CKD were divided into 6 groups according to the National Kidney Foundation classification. Prooxidant status was assessed by assaying thiobarbituric acid reactive substances, hydroperoxides, and protein carbonyls. Antioxidant defence was performed by analysis of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, vitamin E, Iron, and bilirubin. TBARS and LPO were higher in HD patients compared to other groups (), while protein carbonyls were more increased in PD patients. The antioxidant enzymes were declined already at severe stage of CKD and they were declined notably in HD patients (). Similar observation was found for vitamin E, Fe, and bilirubin where we observed a significant decrease in the majority of study groups, especially in HD patients (). The evolution of CKD was associated with elevated OS. HD accentuates lipid, while PD aggravates protein oxidation. However, the activity of antioxidant enzymes was altered by impaired renal function and by both dialysis treatments. Hadja Fatima Tbahriti, Abbou Kaddous, Malika Bouchenak, and Khedidja Mekki Copyright © 2013 Hadja Fatima Tbahriti et al. All rights reserved. Acetylcholinesterase Biosensors for Electrochemical Detection of Organophosphorus Compounds: A Review Mon, 09 Dec 2013 15:38:40 +0000 The exponentially growing population, with limited resources, has exerted an intense pressure on the agriculture sector. In order to achieve high productivity the use of pesticide has increased up to many folds. These pesticides contain organophosphorus (OP) toxic compounds which interfere with the proper functioning of enzyme acetylcholinesterase (AChE) and finally affect the central nervous system (CNS). So, there is a need for routine, continuous, on spot detection of OP compounds which are the main limitations associated with conventional analytical methods. AChE based enzymatic biosensors have been reported by researchers as the most promising tool for analysis of pesticide level to control toxicity and for environment conservation. The present review summarises AChE based biosensors by discussing their characteristic features in terms of fabrication, detection limit, linearity range, time of incubation, and storage stability. Use of nanoparticles in recently reported fabrication strategies has improved the efficiency of biosensors to a great extent making them more reliable and robust. Vikas Dhull, Anjum Gahlaut, Neeraj Dilbaghi, and Vikas Hooda Copyright © 2013 Vikas Dhull et al. All rights reserved. Ameliorative Effects of Acacia Honey against Sodium Arsenite-Induced Oxidative Stress in Some Viscera of Male Wistar Albino Rats Tue, 03 Dec 2013 15:08:50 +0000 Cancer is a leading cause of death worldwide and its development is frequently associated with oxidative stress-induced by carcinogens such as arsenicals. Most foods are basically health-promoting or disease-preventing and a typical example of such type is honey. This study was undertaken to investigate the ameliorative effects of Acacia honey on sodium arsenite-induced oxidative stress in the heart, lung and kidney tissues of male Wistar rats. Male Wistar albino rats divided into four groups of five rats each were administered distilled water, Acacia honey (20%), sodium arsenite (5 mg/kg body weight), Acacia honey, and sodium arsenite daily for one week. They were sacrificed anesthetically using 60 mg/kg sodium pentothal. The tissues were used for the assessment of glutathione peroxidase, catalase, and superoxide dismutase activities, protein content and lipid peroxidation. Sodium arsenite significantly suppressed the glutathione peroxidase, catalase, superoxide dismutase activities with simultaneous induction of lipid peroxidation. Administration of Acacia honey significantly increased glutathione peroxidase, catalase, and superoxide dismutase activities with concomitant suppression of lipid peroxidation as evident by the decrease in malondialdehyde level. From the results obtained, Acacia honey mitigates sodium arsenite induced-oxidative stress in male Wistar albino rats, which suggest that it may attenuate oxidative stress implicated in chemical carcinogenesis. Muhammad Aliyu, Sani Ibrahim, Hajiya M. Inuwa, Abdullahi B. Sallau, Olagunju Abbas, Idowu A. Aimola, Nathan Habila, and Ndidi S. Uche Copyright © 2013 Muhammad Aliyu et al. All rights reserved. Prooxidant-Antioxidant Balance in Umbilical Cord Blood of Infants with Meconium Stained of Amniotic Fluid Thu, 28 Nov 2013 11:37:49 +0000 Objective. Using a novel assay termed prooxidant-antioxidant balance (PAB) assay to determine prooxidant-antioxidant balance in umbilical cord blood of infants with meconium stained of amniotic fluid (MSAF). Passage of meconium in amniotic fluid is associated with increase of neonatal mortality and morbidity. This complication occurs in about 15% of infants and is more widespread in postterm neonates. About 1.5 percent of neonates with MSAF develop meconium aspiration syndrome. Method. Sera of 29 umbilical cord blood of infants with MSAF and 32 healthy infants (HI) were collected. Both groups had nonsmoker and non-alcoholic mothers with no diseases. The PAB was measured. Result. There was a significant increase of PAB value ( HK) in umbilical cord blood of infants with MSAF in comparison to HI ( HK) (). There was no significant correlation between PAB value and age of mothers. Conclusion. The increased PAB value in infants with MSAF showed that these infants are exposed to oxidative stress. Further research with larger population is needed to demonstrate the oxidative stress in infants with MSAF. Mohammad Hassan Arjmand, Farhat Ahmad Shah, Masoud Saleh Moghadam, Fatemeh Tara, Amin Jalili, Mojtaba Mosavi Bazaz, and Daryoush Hamidi Alamdari Copyright © 2013 Mohammad Hassan Arjmand et al. All rights reserved. Long-Term Use of Probiotic-Containing Yogurts Is a Safe Way to Prevent Helicobacter pylori: Based on a Mongolian Gerbil's Model Thu, 21 Nov 2013 15:39:21 +0000 Background. The suppression of Helicobacter pylori (H. pylori) decreases H. pylori-related diseases. The probiotics have an inhibitory effect on H. pylori. Aim. We investigated the effects of long-term use of yogurt on H. pylori based on Mongolian gerbils’ model. Materials and Methods. Yogurt (containing a supplement of Lactobacillus acidophilus, Bifidobacterium lactis, etc.) was used. Forty-six gerbils were divided into five groups. All groups were inoculated with H. pylori for 5 to 8 weeks. The yogurt was given as follows: Group (Gr.) A: from 1st to 4th week; Gr. B from 5th to 8th week; Gr. C: from 17th week to sacrifice; Gr. D: from 5th week to sacrifice. Gerbils were sacrificed on the 52nd week. Histology was evaluated according to the Sydney system. Results. The positive rates of H. pylori were 60% (Gr. A), 75% (Gr. B), 67% (Gr. C), 44% (Gr. D), and 100% (Gr. E). Gr. D showed lower inflammatory score. Only Gr. E (60%) had intestinal metaplasia. Gr. D showed higher IL-10 and lower TNF-α expression than Gr. E. Conclusion. Long-term intake of yogurt could decrease H. pylori infection. The long-term use of yogurt would be an alternative strategy to manage H. pylori infection. Chao-Hung Kuo, Sophie S. W. Wang, Chien-Yu Lu, Huang-Ming Hu, Fu-Chen Kuo, Bi-Chuang Weng, Chun-Chieh Wu, Chung-Jung Liu, Pei-Yun Tsai, Tsung-Cheng Lee, Li-Wei Chen, Kuang-Hung Cheng, Lin-Li Chang, and Deng-Chyang Wu Copyright © 2013 Chao-Hung Kuo et al. All rights reserved. Determination of Coverage Interval of Antioxidant Vitamins (Vitamin C & Vitamin E) in Plasma and Serum of Bengali Population Thu, 14 Nov 2013 18:17:47 +0000 Reference interval of all haematological and biological analytes should be measured for every population because of the huge diversity in genetic make-up, dietary habits. The coverage interval of antioxidant vitamins (vitamin C and α-tocopherol) in the plasma and serum of reference Bengali population was determined and compared with the reference intervals of antioxidant vitamins in the established literature. Adult healthy volunteers from 18 to 68 years of age underwent extensive clinical and investigational procedure and were included in the study. Vitamin C and α-tocopherol were estimated using simple Spectrophotometric method. Of the 71 healthy Bengali volunteers participated, 31 were males and 40 were females. The mean concentration of plasma vitamin C was found to be 0.65 mg/dL. The mean α-tocopherol was found 6.35 mg/L (14.74 μmol/L) in the study population higher than the normal threshold value for α-tocopherol but lower than other populations. The study data enabled us to determine the gender nonspecific coverage interval of antioxidant vitamins, and the intervals were lower than the established reference interval in other populations. Sanghamitra Chakraborty and Indranil Chakraborty Copyright © 2013 Sanghamitra Chakraborty and Indranil Chakraborty. All rights reserved. Inhibition of Rat Muscle and Liver Phosphofructokinases by High Doses of Ethanol Wed, 13 Nov 2013 13:33:01 +0000 Activities of both rat muscle and liver phosphofructokinases are significantly inhibited after a single ethanol intake in the dose of 2.5 g per kg of body weight. This inhibitory effect is indirect, since ethanol in concentration (50 mM) close to that established after 2.5 g per kg of body weight intake cannot decrease their activities in vitro. Inhibition of liver phosphofructokinase activity after the 5.0 g per kg ethanol intake may be direct, since liver phosphofructokinase activity decreases in vitro when ethanol is added to supernatants of rat liver tissue in 100 mM concentration. According to the results of molecular docking, ethanol at high concentrations can be bound by adenine-binding pocket of the allosteric ADP-binding site of liver phosphofructokinase (Asp543, Phe308, Phe538, and Phe671) and its activation by ADP can be blocked by C2H5OH molecule. Direct inhibition of muscle phosphofructokinase activity, probably due to the binding of ethanol to the similar ADP-binding site, is possible when the concentration of ethanol (500 mM) is much higher than the level which can be established in living cells. So, inhibition of muscle phosphofructokinase activity after a single 5.0 g per kg intake is indirect and probably linked with the inhibition of the enzyme by elevated citrate and phosphoenolpyruvate levels. Sergey Vladimirovich Lelevich, Vladislav Victorovich Khrustalev, and Eugene Victorovich Barkovsky Copyright © 2013 Sergey Vladimirovich Lelevich et al. All rights reserved.