American Journal of Biochemistry and Molecular Biology

No Issue Number

• Compositional Analysis α-Lactalbumin
  This review is about the α-Lactalbumin (α-LA), the major whey protein found in the milk of all mammals. It is a simple Ca ²⁺ binding milk protein, homologous in sequence to the lysozyme family and has significant role in biosynthesis of lactose in the lactating mammary gland. The structural and functional analyses so far obtained about this protein were outlined here. As regards the structure, it displays the best-characterized molten globule state which has been studied by various techniques such as X-ray scattering, NMR, thermodynamic study, ultrasonic study etc. The functional aspect shows many remarkable features of its calcium binding sites and is found to be a valuable constituent to enrich the infant formula. In addition, α-LA can alter its biological function depending on the conformational state and actively interact with lipid membranes, which leads to its antimicrobial and antitumor activity that has a vital role to induce apoptosis in tumor cells. Recently it was found that fabrication of α-LA nanoparticles to use in drug and food delivery system in nanomedicine is also possible.
  
• Biology and Partial Sequencing of an Endophytic <I>Fusarium oxysporum</I> and Plant Defense Complex
  The capacity of a non-pathogenic strain of Fusarium oxysporum (IMI 386351) to control fusarium wilt of a susceptible tomato ( Lycopersicon esculentum Mill.) cultivar Danish Export caused by Fusarium oxysporum f. sp. lycopersici was examined along with the biology of that strain, including its phylogenetics. Based on the morphological and the molecular examinations, F . oxysporum IMI 386351 appeared to be a member of the F . oxysporum complex; phylogenetic trees inferred independently and by combining the two partial sequence genes (a mitochondrial small subunit ribosomal DNA and the nuclear translation elongation factor 1α) suggest that this strain might be a member of Fusarium oxysporum f. sp. melonis . The results of seed-coating experiments using five doses of F. oxysporum IMI 386351 revealed a complex three-organism interaction (host-endophyte-pathogen). Between two of the doses, a dose-dependent shift from one form of systemic induced resistance to a distinctly different form apparently occurred. Corresponding with this shift, the pathogen shifted from being virulent to being benign and disease incidence plummeted to zero. A plant growth promoting effect of F. oxysporum IMI 386351, which was probably associated with systemic induced resistance, consistently increased with dose. This study may be the first to demonstrate that, by means of systemic induced resistance, an endophytic fungus (as opposed to a bacterium) can suppress disease symptoms caused by plant pathogenic fungi.
  
• Biochemical Analysis of Black and White Sesame Seeds from China
  An analytical comparison of the biochemical composition of Black Sesame (BS) and White Sesame (WS) produced in China was carried out. The aim of the study was to analyze the black and white sesame seeds grown in China and compare their biochemical properties. Various approved methods that have been reported by researchers were used to do the analysis. Gas Chromatography/Mass Spectra (GC/MS) system was used to identify and quantify the fatty acids. Nicolet 360Ft-IR spectrometer was used to determine the Infrared (IR) spectra of WS and BS. Protein for WS was 22.20%; BS 20.82%; fat WS was 52.61% and BS 48.40%; moisture was higher in WS than BS but ash was higher in BS than WS and the amount was significantly different at (p<0.05); carbohydrate was higher in BS than WS. The two colors were good sources of minerals. Vitamins vary in quantity for the two colors, the same was also observed for the sugars. The fatty acids, oleic and linoleic, were the major unsaturated fatty acids while palmitic and stearic were the main saturated fatty acids significantly observed in both samples. Both colors were higher in essential amino acids with the exception of lysine. The IR spectra of WS and BS showed different peak structures and both possess different functional groups at different regions of their spectra. The overall results indicated that WS and BS have different biochemical properties.
  
• Influence of Live Yeast Culture on Milk Production, Composition and Some Blood Metabolites of Ossimi Ewes During the Milking Period
  A feeding trial was conducted to evaluate the influence of live yeast culture ( Saccharomyces cerevisiae ) on milk production, composition, and some blood metabolites of Ossimi ewes during the milking period. The control group (G1) was fed a concentrate mixture (CFM) and hay (H) and grazed twice daily, while the second group (G2) and third group (G3) were fed the same diet supplemented with 3 or 6 g of live yeast culture (Yea Sacc1026), respectively. The treated groups had significantly higher values (p <0.05) for fat corrected milk (FCM) (740, 605, 571 g/day, for G3, G2 and G1 vs, respectively), while the values for milk yield, fat yield and lactose yield were higher (p <0.05) only in G3 compared with G1. Milk yield values were constantly higher in G3 than in G1 while the values for the G3 were more variable during milking. Milk composition was not significantly affected by yeast supplementation with the exception of urea values which were significantly (p <0.05) lower in G3. Yeast administration influenced β-Hydroxy-Butyrate (BHB) values, which were significantly (p <0.05) higher in the treated groups; and non-esterified fatty acids (NEFA) values, which were significantly (p <0.05) higher only in the G3 compared with the G1. Other blood metabolites values were not influenced by the treatments. It was concluded that supplementation with live yeast culture, under the conditions of this experiment, had a significant effect on the performance and metabolism of Ossimi ewes during the milking period. Based on more constant results, it is recommend including live yeast culture (Yea Sacc1026) at 6 g/animal/day as appropriate level for field conditions.
  
• Biochemical Studies of Iron Fortified Nigerian Rice Fed to Phenylhydrazine Induced Anaemic Rats
  The blood chemistry, hematological and oxidative stress parameters of anaemic rats fed iron-fortified Nigerian Ofada rice with Ferrazone ^® (NaFeEDTA) were investigated. Forty male rats were divided equally into 4 groups. Groups 1 and 2 were induced with anaemia using phenylhydrazine and were fed with and without iron fortified diet respectively, while groups 3 and 4 which served as control were non-anaemic and were fed with and without iron fortified diet respectively, for 30 days. The blood chemistry, electrolyte level, hematology, anti-oxidant enzyme activities and tissue histology were determined. Packed cell volume , hemoglobin level and red blood cell count were significantly (p<0.01) increased in rats fed with/without iron fortified diet as compared to the control. The activities of aspartate amino transferase, alanine amino transferase, direct bilirubin, creatinine and triacylglycerol levels were significantly increased in anaemic rats fed with iron fortified diet as compared with the control. There was a significant decrease in superoxide dismutase (SOD) activity and malonyldialdehyde (MDA) level of anaemic rats fed with fortified diet as compared with the control group, while there was a significant increase in SOD and catalase activities and a decrease in MDA level of anaemic rats fed with unfortified diet. There was a significant decrease in sodium and potassium levels and increased in the brain iron levels of anaemic rats fed with iron fortified diet as compared with the control. Data of the study show that iron fortification using Ferrazone ^® affected the hematology, lipid peroxidation, electrolytes and blood chemistry of rats.
  
• Genetic Diversity in <I>Salix viminalis </I> in the Kashmir Valley, India
  In Kashmir, Salix is cultivated on large scale. There are diverse cultivars of Salix found in Kashmir Valley but they are not well demarked due to similarities in certain morphological characteristics. Salix species is an important source of aspirin and salicylic acid. It has tremendous economic importance. Its wood is used for manufacturing different types of furniture, bats etc. Therefore, in the present study a fingerprinting technique has been used to differentiate cultivars of Salix found in Kashmir Valley. Among Salix cultivars, genetic diversity has been ascertained in the S. viminalis using Amplified Fragment Length Polymorphism (AFLP) technique. AFLP analysis of different Salix viminalis cultivars which were difficult to identify morphologically revealed that they are different cultivars of the same species. AFLP markers were able to reveal that the four Salix cultivars taken for study were highly diverse at genetic level. The 4 primers used generated a total of 240 bands of which 197 (82%) were polymorphic. The broad genetic base can be attributed to their out-crossing nature. Our study may be useful in identifying diverse genetic stock of S. viminalis , which may be conserved on priority basis.
  
• Quaternary Structure of Omp85/YaeT/BamA of <I>Yersinia pestis</I>
  The aim of the study was to investigate quaternary structure of Omp85 outer membrane protein of Y. pestis , because of its potential use as a potential therapeutic target. Bacterial outer membrane proteins are suitable targets for the antimicrobial drugs and vaccine development. In Gram-negative bacteria, outer membrane protein, Omp85 is an integral part of the protein machinery. Inhibiting and disabling of Omp85 has a negative effect on the secretion of virulence factors, as well as a bactericidal effect. Previous research has identified components of the Omp85 complex and has started to yield structural insights into Omp85 family proteins, the quaternary structure of Omp85 itself remains under investigation. We present a quaternary structure study of Yersinia pestis Omp85 in vitro and in vivo using multiple approaches. Full-length Omp85, purified without denaturation, is shown to form monomers, dimers and tetramers in vitro using cross-linking and sedimentation studies. In vivo studies, using Western blot of outer membranes resolved on semi-native LDS-PAGE, indicate that Omp85 formed tetramers. Our studies contribute to a better understanding of Omp85 structure and mechanism. The applied relevance of this study has potential use in the development of new vaccines and antimicrobials, which may be potent weapons in the fight against bacterial infections.
  
• Comparative Study of RAPD and ISSR Markers to Assess the Genetic Diversity of Betel Vine (<I>Piper betle</I> L.) in Orissa, India
  In Orissa Betel vine ( Piper betle L., family Piperaceae) is an important asexually propagated cash crop comprising of several cultivars. There are many cultivares but they are not well demarcated due to similarities in the morphological characters and in certain places same cultivars are cultivated under different local name. Therefore, in the present study DNA fingerprinting technique has been used to differentiate cultivars of betel vine for crop improvement programme. So Comparative study of both RAPD and ISSR markers analysis were used to establish genetic identities and evaluate genetic diversity among fifteen cultivars of betel vine grown in different parts of Orissa. Thirty RAPD and 25 ISSR primers were tested to resolve the genetic diversity among the cultivars. Twenty RAPD and 18 ISSR primers resulted in 523 amplicons. Out of these 504 were polymorphic loci and 54 were found to be unique. The extent of genetic diversity and relatedness among 15 cultivars were computed through Jaccard’s similarity coefficient. Maximum similarity (0.68) was observed between Balipana and Birkoli and minimum (0.114) for Banglamandesore chitalpudi and Halisahar Sanchi. All the cultivars were related with each other with an average similarity of 0.2913. Dendrogram showed Godibangala was separated from rest of the species into isolated clade in both the analysis. Correlation between RAPD and ISSR marker was very low (r = 0.17). RAPD showed high correlation with all the primers.
  
• Effects of Silibinin on Hair Follicle Stem Cells Differentiation to Neural-like Cells
  The hair follicle stem cells are an abundant and easily accessible source of pluripotent adult stem cells. Hairfollicle stem cells differentiate into neurons and glial cells in vitro and express stem cell markers such as tubulin, RIP, P75, and S100. Silibinin, a flavonolignan and the active component of Milk thistle ( Silybum Marianum ) has been used as a dietary supplement and herbal medication. The aim of the present study was to investigate the effects of Silibinin on hair follicle stem cells differentiation to neural-like cells. The bulge region of the rat whisker was isolated and cultured in Dulbecco’s modified Eagle’s medium supplemented with different concentrations of silibinin and Neurotrophin-3. One week later , bulge cells immunostained with Nestin and p75 and then with tubulin . The morphological and biological features of cultured bulge cells were observed by light microscopy. Present data demonstrated that silibinin (0.5 μg mL ^-1 ) can promote differentiation of hair follicle stem cells to neural-like cells.
  
• Genetic Diversity of Persian Wheat (<I>Triticum turgidum</I> ssp. <I>carthlicum</I>) Accessions by EST-SSR Markers
  To estimate the allelic variation at the expressed sequence tag simple sequence repeats (EST-SSR) among Persian wheat ( Triticum turgidum ssp. carthlicum L.) accessions collected from fifteen countries and provide information for wheat breeding and improvement in southwest China, 87 Persian wheat accessions from fifteen countries were investigated by using EST-SSR markers. EST-SSRs were molecular markers belonging to the transcribed region of the genome. Therefore, any polymorphism detected by EST-SSRs might reflect better relationship among species or varieties. Fourteen primer pairs could successfully amplify the fragments in the 87 accessions, of which tri-nucleotide repeats were the dominant type. A total of 33 eSSR alleles were detected, and the number of alleles detected by single pair primers ranged from 3 to 7 per locus, with an average of 3.71. Clustering analysis suggested that most of the accessions with adjacent geographic origins had the tendency to cluster together. Therefore, when used in Persian wheat genetic analysis, EST-SSR markers not only act as genetic markers but also reveal differences in related gene expression.
  
• Detection of Arg 72Pro Polymorphism of the Tumor Suppressor Gene (<I>TP53</I>) by a Rapid One-step Tetra-primer Amplification Refractory Mutation System-PCR
  The Tumor Protein p53 gene ( TP53 ), known as the guardian of the genome, encodes the p53 protein that plays an important role in cell cycle and in maintenance of the genome stability by preventing mutations. The transition from G to C at codon 72 of the TP53 gene represents a non-synonymous single nucleotide polymorphism (Arg 72Pro) that has been extensively genotyped for association with a wide variety of cancers. A new method for Arg 72Pro detection is described based on the tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR), with a single PCR to discriminate both alleles. Two primers that amplify a common amplicon independently of the allele considered are combined in a single PCR reaction with two specific primers, differing in the 3U base. In the presence of Pro/Pro or Arg/Arg, amplification occurs both in the general amplicon and in the specific allele; in the presence of Arg/Pro three different amplicons are produced. Validation by PCR-RFLP revealed 100% accordance for genotype assignment. The method was successfully applied for genotype determination of the Arg 72Pro in a novel population from Aleppo, Syria, with 51% homozygous for the Arg allele and 12% homozygous for the Pro Allele.
  
• Evaluation of Toxicological and Standardization Parameters and Phytochemical Investigation of Ficus deltoidea Leaves
  Medicinal herbs were the primary health care agent over the many centuries before development of modern medicine. The fact that herbal medicines have been employed for such a long time does not guarantee their efficacy and safety. On that basis, an attempt was made on efficacy and safety studies of a well known medicinal herb in Malaysia, Ficus deltoidea by evaluation of toxicological parameter like heavy metals and standardization parameters like, physical constants, ash content, Microbial Limit Test (MLT) and screening phytoconstituents are present in the leaves of F. deltoidea . Heavy metals analysis was done by atomic absorption spectrometry. Physicochemical determinations, including moisture, volatile and total ash content were carried out by Thermogravimetric Analyzer. Microbial Limit Test was done as per the United State Pharmacopoeia method. Thin layer chromatography was carried out for phytochemical screening with normal silica plate using various chemical reagents. The contents of Cd, Pb and As were found to be 0.069, 0.761 and 0.422 ppm, respectively. While Hg was not detected in F. deltoidea leaves. MLT test showed 5.0x10 ⁶ and 3.3x10 ⁷ cfu g ^-1 for Total aerobic microbial count and total combined mold and yeast respectively. However, Staphylococcus aureus , Pseudomonas aeruginosa , Salmonella sp. and Escherichia coli were found to absent . Phytochemical studies of petroleum ether, chloroform, methanol and water extracts showed presence of saponin, amino acid, flavonoids and terpenoids. Hence, there is an urgent need for mandatory evaluation of these parameters in every crude drug before further processing to ensure safety and efficacy of medicinal plants. In conclusion, it was shown that the extracts of F. deltoidea leaves were enriched with chemically diversified phytoconstituents which could be useful for various pharmacological activities.
  
• Chemical Assistance in Refolding of Bacterial Inclusion Bodies
  Escherichia coli is one of the most widely used hosts for the production of recombinant proteins but insoluble expression of heterologous proteins is a major bottleneck in production of recombinant proteins in E. coli . In-vitro refolding of inclusion body into proteins with native conformations is a solution for this problem but there is a need for optimization of condition for each protein specifically. Several approaches have been described for in vitro refolding; most of them involve the use of additives for assisting correct folding. Co-solutes play a major role in refolding process and can be classified according to their function as, aggregation suppressors and folding enhancers. This study presents a review of additives that are used in refolding process of insoluble recombinant proteins in small scale and industrial processes.
  
• Bioactive Compounds from Endophytes and their Potential in Pharmaceutical Effect: A Review
  Endophytes are microorganisms that live inside the host plant tissues which have novel metabolites exhibiting a variety of biological activities against different diseases. In fact, a significant number of interesting natural bioactive compounds have been reported in the last years. The microbial biotransformation process is a novel alternative method to obtain bioactive compounds. In this review, some aspects in the phytochemistry of endophytes producing Phytochemicals and its pharmaceutical effects are discussed.
  
• Serum Copper and Vascular Endothelial Growth Factor (VEGF-A) in Dysfunctional Uterine Bleeding
  Over the past 20 years there has been an upsurge of interest proper cause of abnormal angiogenesis of DUB. This study particularly focused on the association of serum copper and vascular endothelial growth factor (VEGF-A) in patients with Dysfunctional Uterine Bleeding (DUB). Blood samples were collected from female patients suffering with DUB (n = 50) as well as healthy females as controls (n = 50). Serum copper levels were estimated by spectrophotometric method and serum VEGF-A by ELISA technique and compared with ultrasonographic measurement of endometrial thickness in both patients and controls. A significant increase in serum copper levels and an almost two fold increase in serum VEGF-A was observed in DUB patients when compared with controls. Correlation(r) between serum VEGF-A levels and endometrial thickness was 0.97. Odd Ratio for copper, VEGF-A and combination of copper and VEGF-A was 0.0426, 0.0947 and 0.0313, respectively, in all these cases odds ratio was less than one. The abnormal angiogenesis in DUB could be due to increased serum copper levels, which in turn stimulates factors like VEGF-A, thereby causing an increase in endometrial growth.
  
• Single Cell Gel Electrophoretic Analysis of 2-Aminoanthracene Exposed F-344 Rats
  To assess DNA damage from exposure to 2-Aminoanthracene (2AA), Single Cell Gel Electrophoresis (SGCE) was employed to examine single strand breaks in blood of Fisher-344 rats. Although subcellular injury due to 2-AA exposure has been noted in the past, there is yet to be a direct demonstration of genetic alterations due to 2-AA intoxication. In the current study, alkaline comet assay was used to evaluate the type of DNA damage that is on-going in the blood samples of F-344 animals. The animals were fed control (0 mg kg ^-1 ), LD (50 mg kg ^-1 ), MD (75 mg kg ^-1 ) and HD (100 mg kg ^-1 ) 2-AA diet for two and four weeks. At the end of each exposure period (14 or 28 days), rats were euthanized with CO ₂ and blood was collected by cardiac puncture. Twenty micro liters of whole blood samples were added to 1 mL of Hanks Balanced Salt Solution (HBSS without Ca ²⁺ and Mg ²⁺ ) and subsequently snapped frozen in liquid nitrogen. Following comet assay analysis, single strand break were assessed via tail moment, tail length, tail intensity and head intensity scoring of comets along with cell viability. There seems to be an apparent dose response. That is 50, 75 and 100 mg kg ^-1 diet 2-AA rats when compared with the control demonstrated significant (p<0.05) damage as measured by tail length and tail moment values. Head and tail intensity values for intoxicated animals were also significant at 2 and 4 weeks relative to the controls, although there were no dramatic shifts between exposure time.
  
• Phylogenetics and in silico Docking Studies Between Coat Protein of Mimosa Yellow Vein Virus and Whey α-lactalbumin
  Begomovirus associated symptoms were observed in several Mimosa pudica plants growing in crop fields of Lakshmangarh, Rajasthan (India). Amplification of a PCR product was found up to the expected size (~570 bp) during agarose gel electrophoresis only in infected samples not in healthy samples. The PCR product was cloned and partially sequenced (GenBank accession HQ876467) and sequence analysis of HQ876467 by using BLASTn revealed the highest 98% nucleotide sequence identities with Ageratum yellow vein china virus (AJ558120). GeneBank-NCBI generated CDC for HQ876467 from 1 to 774 (ADW83735) and it was utilized for in silico analysis. In Neighbor-Joining tree, the coat protein of mimosa yellow vein virus (ADW83735) place in monophyletic clusters of 74 bootstrap values with AV1 protein of Ageratum yellow vein China virus (CAD90081). The closest homologue of coat protein was 2B7R A, with highest sequence identity only 37% that was selected as representative model using homology modeling softwares. Homology modeling study of coat protein (ADW83735) and docking between α-lactalbumin and coat protein was carried out by using modeling and docking software (Hex 6.3). The ADW83735 model was validated by using Procheck server for reliability that results shown only 49.2% of residues present in core region. On the basis of RMS and energy values, the best docking orientation -1.00 was selected. This study will be used for the screening of inhibitors against mimosa yellow vein virus proteins and can be further applied in future antiviral agent designing.
  
• Coagulation Factors Evaluation in NIDDM Patients
  Diabetes mellitus is the major risk factor for vascular diseases presumably by inducing hypercoagulable state leading to thrombotic plaque in small blood vessels. There are miscellaneous reports showing the intervention of some coagulation factors/co-factors, especially I, II, V, VII, VIII, IX, X, XI, vonWillebrand and tissue factor in micro and macrovascular complications. In the present we used 40 uniformed NIDDM patients and 21 healthy volunteers as control for the study. Based on the results obtained for coagulation tests such as PT, APTT and the specific activity or concentration determination for factors I, II, V, VII, VIII, IX, X, X and von Willebrand, we made a new comprehensive comparison between NIDDM and healthy groups. Present results show the major interference of intrinsic and common pathway of coagulation cascade factors and less of extrinsic pathway in hypercoagulability state in NIDDM patients. Present results show that there are significant increase (p<0.05) in plasma activities of coagulation factors, FII, FIX, FXI in intrinsic and common pathways and significant decrease in activity of factor VIII and prolonged prothrombin time in NIDDM patients (n = 40) in contrast to normal controls (n = 21). This finding leads us to conclude that this pattern could be used as diagnostic criteria for the presence of hypercoagulable state and as prognostic tools for susceptibility to vascular disease in NIDDM patients. Second, our findings in conjunction with patients history indicates that the presence of hypercoagulable state is not sufficient to induced vascular problems in NIDDM and instead it need additional triggering factors to initiate vascular thrombotic problems e.g., endothelium damage.
  
• Immobilization of Microbial (Wild and Mutant Strains) Amylase on Coconut Fiber and Alginate Matrix for Enhanced Activity
  Reports are not available for immobilization of on α-amylase produced by Bacillus subtilis, Agrobacterium tumefaciens, Escherichia coli, Xanthomonas campestris, Staphylococcus aureus, Aspergillus niger, Aspergillus flavus, Metarhizium anisopliae, Azotobacter chroococcum and Rhizopus oryzae on coconut coir. Hence, an attempt was made in present study to immobilize the free amylase on coconut coir. Cell bound amylase of Bacillus subtilis, Escherichia coli, Xanthomonas campestris, Aspergillus niger, Aspergillus flavus and Rhizopus oryzae were immobilized by entrapment in calcium alginate. The immobilized enzyme on cells showed higher α-amylase production compared to free cells. The enzyme was characterized by Thin Layer Chromatography (TLC) and confirmed as alpha amylase. A simple and inexpensive method for immobilizing α-amylase on coconut fiber was also adopted. Highest amylase activity was observed in X. campestris on immobilized state on coconut fibers (1.708±0.103 U/min/g) and also in alginate matrix (0.948±0.082 U/min/100 beads). Hence, the cells of X. campestris were mutated with UV (for different time intervals: 1, 2, 4 and 8 min) and acridine orange (in different concentrations: 1, 2, 4 and 8 μg mL ^-1 ). The immobilized catalyst of mutant strain of X. campestris AO ₄ on alginate matrix exhibited 66.31% (4.125±0.056 U/min/100 beads) higher than that of the parent strain. It exhibited highest activity, when immobilized on coconut coir. Thus the mutant strain of X. campestris AO ₄ could be used in industries for the efficient conversion of starch to oligosaccharides.
  

  No Issue Number

• Induction and Uncoupling of Rat Liver Mitochondria by Oral Administered Coartemether
  This study investigated the cytotoxic effect of varying doses of antimalarial coartemether (2.0, 4.0, 8.0 and 10.0 mg kg ^-1 ) in the presence of ferrous sulphate (2800 mg kg ^-1 ) for 3 days on normal rat liver mitochondrial membrane permeability transition pore opening and F ₁ F ₀ ATPase activity. Swelling was estimated spectrophotometrically under succinate energized condition. Calcium ion treated mitochondria preloaded with coartemether induced swelling in a concentration dependent manner in vitro . Swelling was amplified in the presence of ferrous. Coartemether alone and combined coartemether-ferrous also induced mitochondrial swelling in the presence of spermine. In vivo study further showed that 10.0 mg kg ^-1 coartemether induced swelling in the presence of ferrous. Coartemether stimulated an increased activity of mitochondrial F ₁ F ₀ ATPase in a concentration dependent manner. Thus, these findings indicate that coartemether at high dose in the presence of ferrous sulphate could be an inducer of mitochondrial mega pore opening and an uncoupler of oxidative phosphorylation initiating apoptosis.
  
• Purification and Physicochemical Characterization of Anti-Gardnerella vaginalis Bacteriocin HV6b Produced by Lactobacillus fermentum Isolate from Human Vaginal Ecosystem
  Anti -Gardnerella vaginalis, Lactobacillus fermentum bacteriocin HV6b producing strain was isolated from vaginal swabs of healthy, fecund females. It was identified, purified and further characterized physicochemically. The relative molecular mass of bacteriocin was found to be less than 7 kDa. Maximum bacteriocin production was observed at 37°C, pH 6.5 after 16-18 h of incubation Proteinase K and pepsin strongly inhibited bacteriocin production. The bacteriocin has purified by absorption desorption method and characterized by reverse phase liquid chromatography. It exhibited high thermal stability and showed inhibitory activity over wide range of pH. The study revealed the possibility of using bacteriocin for bio-medical use and the L. fermentum strain as probiotic.
  
• Anti-inflammatory and Antioxidant Activities of a Fraction I1 of Male Inflorescences of Borassus aethiopum Mart (Arecaceae)
  In previous studies, using a bioassay-guided fractionation procedure; five fractions (E1F1, E2F2, E3F3, E4F4 and E5F5) from powdered Borassus aethiopum male inflorescences were extracted. Among these, the dichloromethane methanol extract E2F2 was found to exert significant anti-inflammatory and antipyretic activities and pro apoptotic effect. It seemed important to pursue the investigations to understand the mechanism of the anti-inflammatory activity. The anti-inflammatory activity was studied, C Reactive Protein (CRP) level in mice blood was immunoturbidimetry after inflammation induction and antioxidant activity was studied using 1,1 diphenyl picrylhydrazyl (DPPH). Phytochemical screening was carried out according to the methodology for chemical analysis for vegetable drugs. Among 3 fractions (I1; I2, I3) of E2F2, I1 was the most active with a percentage of inhibition (PI) of 80%. This anti-inflammatory activity was twice high than indometacin (PI = 40%). The I1 fraction cause significant decline of concentration of CRP compared with indometacin. The radical scavenging activities of I1 were approximately 4 times lower than ascorbic acid. Phytochemical analyses of Borassus aethiopum extracts revealed the presence of terpenoids, steroids and saponins which all have been shown to be potent anti-inflammatory and antioxidants. The present study confirmed the anti-inflammatory and antioxidant potential of Borassus aethiopum extracts with results comparable with those of standard compounds such as indometacin. Further studies are needed to isolate, purify and identify the chemical structure of the compounds responsible for anti-inflammatory and antioxidant activity.
  
• Effect of Quercetin and Epicatechin on the Transcript Expression and Activity of Antioxidant Enzymes in Tobacco Seedlings
  The aim of this study was to see the effect of flavonoids quercetin and epicatechin on the transcript expression and activity of antioxidant enzymes. For this, tobacco seedlings were exposed to 50 and 100 μM quercetin and epicatechin for 21 days. The transcript expression level of various antioxidant enzymes like Glutathione-S-Transferase (GST), Glutathione Peroxidase (GPx), Ascorbate Peroxidase (APx), Glutathione Reductase (GR), Catalase (CAT) and Superoxide dismutase (SOD) were monitored in the root and shoot of exposed seedlings through reverse transcriptase PCR. The activity of these enzymes was studied spectrophotometrically. Interestingly, 50 μM epicatechin and 100 μM quercetin exposures were found to increase the expression of genes encoding antioxidant enzymes in shoot. In tobacco root, only Glutathione-S-Transferase (GST) and GPx expression was increased with 50 μM epicatechin and 100 μM quercetin exposures. Activity assay of all the enzymes showed similar trend to that of the transcript expression in shoot tissue. While in root, except CAT and SOD other enzymes activity also showed similar trend to that of expression pattern. Results have suggested the possible regulation of antioxidant enzymes by these two flavonoids at transcriptional and post-transcriptional level. Additionally, appropriate levels of such flavonoids seem to be essential for such regulations.
  
• XRD and Physicochemical Evaluation of Hibiscus sabdariffa Cellulose-Butyl Acrylate-co-vinyl Monomer Graft
  Utilizing the renewable waste biomass to procure advanced materials has been the aim of research. Various reaction parameters were optimized for the graft co-polymerization of Hibiscus sabdariffa cellulose fiber using binary vinyl monomeric mixture. The graft co-polymers thus obtained were characterized by XRD technique and the results received the supporting evidence by various other advanced analytical techniques. The percentage crystallinity and crystallinity index were found to decrease with increase in grafting while a reduction in moisture absorption was observed. There was an increase in physico-chemico-thermal resistance in the graft copolymers. These graft copolymer can be used in various scientific applications.
  
• Competitiveness of Polymerase Chain Reaction to Alternate Amplification Methods
  PCR is the first method developed for amplification of DNA/nucleic acids in vitro . It was the method of choice since it's invent. PCR has many limitations which gave birth to alternative methods such as Loop Mediated Isothermal Amplification (LAMP), Nucleic Acid Sequence Based Amplification (NASBA), Self Sustained Sequence Replication (3SR), Rolling Circle Amplification (RCA) etc. A comprehensive review of the literature on the principles, limitations, challenges, improvements and alternatives of PCR was performed. Though PCR encountered some limitation afterwards and a no of technically improved alternatives has been devised worldwide, it still holds its standings as a gold standard for nucleic acid amplification preventing it from being obsolete. PCR has gone through some phenomenal modifications making continuous improvement of this method. Although, latter techniques offer better sensitivity and advantages where PCR has limitations, it is still commonly applied in the field of molecular biology for its extensive information all over the world, availability of reagents and methods, widespread validation and very good technical understanding among researchers. PCR is not obsolete despite of threats faced from other methods; rather it is continuously being modified to overcome the limitations. It will remain one of the ultimate methods of choice especially in developing countries for long.
  
• Chemical Analysis of Essential Oils of Some Syrian Wild Iris Species
  This research was aimed at investigating qualitative and quantitative variation among oils of some Syrian wild Iris species compared to those of Iris germanica . Hydrodistillation was used to extract the essential oil from different wild Iris rhizomes. 0.1- 0.2% of oil yield was obtained. Gas Chromatography-mass Spectrometry (GC-MS) analysis of the essential oil have indicated the presence of 23 compounds in Iris germanica, Iris barnumae, Iris bostrensis and 19 in Iris aurantiaca . Results were statistically analyzed by the SPSS computer program. The major compound in these essential oils was myristic acid (61.42, 70.67, 51.15 and 79.67%) in Iris germanica, Iris aurantiaca, Iris barnumae and Iris bostrensis, respectively. The other sub major compounds obtained were lauric acid, decanoic acid (capric acid), palmitic acid methyl ester, octadecanoic acid methyl ester, elaidic acid methyl ester (9-octadecenoic acid methyl ester (e)-) and palmitic acid. This is the first report on isolation and identification of oil from Syrian wild Iris plants by GC-MS.
  
• L-arginine Exposure Improves Renal Function Markers of Metabolic Syndrome in Female Rats
  Female gender is an independent risk factor for the development of metabolic syndrome (MES) (a cluster of features indicating metabolic disorders), that is associated with kidney damage, insulin resistance and a significant reduction in Nitric Oxide (NO), a major metabolite of L-arginine (ARG). This study aimed to ascertain the effect of ARG on selected markers of MES related to kidney damage in female Wistar albino rats. Two groups of rats were given 3 mL kg ^-1 body weight (b.wt.) of distilled water, DW and 60 mg kg ^-1 b.wt. of ARG, respectively as control and treated groups. Exposing the female rats to ARG caused a significant decrease (p <0.01) in the concentration of urea (6.34±0.23 mg/100 mL), creatinine (4.41±0.50 mg/100 mL) and albumin (14.30±0.15 mg/100 mL) in rats’ serum. It decreased (p <0.01) creatinine clearance (1.78±0.27 mL min ^-1 ) but elicited a significant increase (p <0.01) in the albumin:creatinine ratio (3.27±0.32) of the rats. Improved kidney histology as indicated by lots of renal corpuscles, was observed in the ARG-fed group while correlation analysis showed that urea correlated positively (r = 0.01) with creatinine, albumin and creatinine clearance, but negatively (r = 0.01) with Albumin: Creatinine ratio. The study suggests that L-arginine ingestion could improve these renal function markers and perhaps, metabolic syndrome related to kidney dysfunction, in female Wistar rats. The effect could be concerted and significant as indicated by the histomorpholgy and correlation results. Thus, with the abundance of ARG in nuts, including walnut, cashew nut, ground nut and even coconut, the implication of this study in the prevention and management of MES in, especially female, animals is noteworthy hence, deserve follow up, probably in humans.
  
• Insights into Steviol Glycoside Biosynthesis Pathway Enzymes Through Structural Homology Modeling
  Steviol glycosides are the major secondary metabolites synthesized through steviol glycoside biosynthesis pathway operating in the leaves of Stevia rebaudiana . Present article documents the structural analysis of enzymes specific to steviol glycoside biosynthesis pathway, kaurenoic acid-13 hydoxylase (KAH) and three UDP-glycosyltransferases (UGT85C2, UGT74G1 and UGT76G1). The in silico protein structure prediction server SWISS MODEL was used to predict and evaluate the models. The secondary structure data of predicted model for KAH was in accordance to that of cytochrome P450s suggesting its nativeness to the respective superfamily. Similarly, the secondary structure data of target UGTs also showed conservation with the structural information of glycosyltransferases superfamily. PROCHECK and QMEAN Z-score evaluations suggested that the models predicted for the 4 query enzymatic proteins were of good quality. In addition, Ligand binding site analysis and molecular docking analysis was carried out for the predicted models. The following data suggested a possibility of the presence of an alternate pathway for the synthesis of steviol glycosides.
  
• Biochemical Mechanisms of Resistance to Black Pod Disease in Cocoa (Theobroma cacao L.)
  Black pod disease caused by Phytophthora palmivora and Phytophthora megakarya is an important disease of cocoa and host plant resistance is one of the important components of management of this disease. Therefore, a diverse array of 12 cocoa genotypes was evaluated to identify biochemical characteristics conferring resistance to black pod disease. Resistance to black pod disease measured as leaf disc scores, pod lesion numbers, pod lesion sizes and natural field infection was associated with amounts of nitrogen, protein, soluble sugars, insoluble sugars, total polyphenols, flavonoids, tannins and lignin in leaf and pod of cocoa. The levels of these biochemical compounds in leaf and pod increased after inoculation with P. megakarya . Principal component analysis shows that 90% of the total variability in the eight original biochemical variables is captured in the first two principal components and 95% in the first three principal components. The first two principal components were defined by healthy pod lignin and healthy pod insoluble sugar and principal component three was mainly defined by healthy leaf protein. Phytochemical characterization of the 12 cocoa genotypes by cluster analysis revealed two major clusters. Cluster one consisted of Imc67, Na33, T79/501, T63/971, Imc53 and Sca9 which were susceptible to black pod and cluster two was made up of Pa150, T60/887, Sca6, Imc76, Pa7/808 and T85/799 which were resistant. Correlation coefficients, multiple and step-wise regressions indicated that insoluble sugar, flavonoid, tannins and lignin were the most reliable biochemical factors and these could be used as marker traits to screen and select for resistance to black pod disease of cocoa.
  
• Modulation of Cardiovascular Risk Factors (Haematological and Haemorrheological Parameters) Caused by Sucrose Diet
  The involvement of sucrose and its amount in the causation of cardiovascular disease is still controversial and inconclusive. The two latest reports of WHO/FAO and Institute of Medicine of Food and Nutritional Board (IOM of FNB) on optimal level of sucrose consumption are at least contradictory; therefore the need to clarify the effect of different concentrations of sucrose consumption on cardiovascular disease risk factor is expedient. Effect of sucrose consumption was assessed on twenty four male albino rats, four to six weeks old, 48-65 g, divided into five groups: G1 (control), G2 (10% energy supply from sucrose), G3 (20% energy supply from sucrose), G4 (30% energy supply from sucrose). The following parameters were determined: red blood cell count, white blood cell count, packed cell volume, blood and plasma viscosities; fibrinogen level and erythrocyte sedimentation rate. Analyses revealed that inclusion of sucrose at concentration of 20% energy supply significantly increased (p<0.05) blood viscosity by 97.59%, plasma viscosity 16.48%, erythrocyte sedimentation rate 40.00%, plasma fibrinogen13.63% and white blood cell count 6.59%, while no significant effect (p>0.05) was observed on red blood cell count and packed cell volume. The study revealed that consumption of sucrose at twenty percent energy supply increased some selected haematological and haemorrheological parameters associated with cardiovascular disease.
  
• Phylogenetic Diversity of Sorghum bicolor (L.) Moench Accessions from Different Regions in Sudan
  Sorghum ( Sorghum bicolor L.) is believed to be domesticated for the first time within the geographical range of Sudan, so, studying the genetic variation of sorghum genotypes collections attracts special interest. In this study, 17 sorghum accessions with important agronomic traits, representing four states in Sudan were assayed for polymorphism using Random Amplified Polymorphic DNA technique (RAPD). Ten primers out of 40 tested (A-1, B-20, C-20, D-18, OPE-04, UBC-101, UBC-103, UBC-127, UBC-155 and UBC-157) showed high polymorphism among the accessions. The results indicated 110 polymorphic bands out of 145 bands with percentage of polymorphic bands of 77.6%. Unweighted Pair Group Method with Arithmetic Mean (UPGMA) result showed two major clusters, with a clear trend of grouping of each region. A cluster contained all accessions from eastern Sudan (Red Sea). However, accessions from north Sudan (River Nile), western Sudan (North Kordofan) and south Sudan (Blue Nile) were diverse in another cluster. Based on the results of this study RAPD technique proved to be useful to study genetic variation among the Sudanese sorghum accessions.
  
• Assay of Glucose 6-phosphate Dehydrogenase Enzyme and its Correlation with Disease Prevalence in Patients with Plasmodium falciparum Malaria
  Falciparum malaria is a major global health problem and is third leading cause of death after HIV and tuberculosis. Although, several targets and related drugs are available, yet the parasite evolves a resistance mechanism to most of the existing drugs. Under such circumstances, it is imperative to explore antimalarial drug targets and effective drugs. Malaria is an interesting case of evolutionary selection and several host genetic factors have been selected in response to Plasmodium falciparum infection, such as hemoglobin variants, glucose-6-phosphate dehydrogenase (G6PD) and pyruvate kinase deficiency. Among these G6PD deficiency is one of the most studied host genetic factor that confer resistance to malaria in endemic and non endemic region. We have studied G6PD deficiency in Falciparum malaria patients and ethnically matched controls in non endemic region of 170 blood samples. G6PD deficiency in samples was detected by Fluorescent spot test. The samples were incubated with reaction mixture, spotted and visualized (366 nm) under ultraviolet light. Results indicate that the frequency of % G6PD deficiency is 9.2 in males and 7.14 in females in non endemic region. On the basis of our data and earlier studies, we conclude that G6PD deficiency is more prevalent in those areas where the frequency of malaria infection is high. G6PD is a selective force against the pressure of malaria in endemic regions while in non endemic regions the burden of malaria is seasonal and low which accounts for the deficiency of enzyme in conferring protection against malaria.
  
• Effect of Mistletoe (Viscum album) Extract on the Blood Glucose, Liver Enzymes and Electrolyte Balance in Alloxan Induced Diabetic Rats
  The effect of aqueous extract of mistletoe on the blood glucose, electrolytes, transaminases and phosphatase activities of alloxan induced diabetic rats was investigated using their respective kits. Twenty male rats, weighing between 90 and 120 g kg ^-1 b.wt. were used. Alloxan was injected into the experimental animals at a dosage of 100 mg kg ^-1 b.wt. The rats were divided into 5 groups: Group 1: non-diabetic received normal rat feeds comprising of clean water and growers mash. Group 2: diabetic control received the same treatment as group 1. Group 3: diabetic received 20% of the extract, Group 4: diabetic, received 40% of the extract while group 5 diabetic rats received 60% of the extract. The duration of the experiment was four weeks. Results indicate that all concentrations of the extract administered reduced the elevated blood glucose levels of the diabetic animals although they could not statistically ameliorate it (p>0.05), administration of 20 and 40% of the extract was most effective in ameliorating the altered K ⁺ concentration of the diabetic rats, while 40 and 60% of the extract proved to be statistically most effective in ameliorating the elevated bicarbonate levels of the diabetic rats (p<0.05). All concentrations administered also ameliorated the altered sodium and alanine transaminase activities of the diabetic animals while they slightly improved their chloride levels (p>0.05). In addition, 60% of the extract significantly ameliorated the altered phosphatase activities and body weights of the diabetic rats while 40 and 60% of the extract significantly ameliorated the altered aspartate transaminase activities of the diabetic animals (p<0.05). It can be deduced that mistletoe has anti-diabetic potentials, in addition to ameliorating the altered electrolyte balance arising from diabetes. Finally, the dietary supplement may protect liver cells from free radical damage and could be effective in treating hepatoctye damage in diabetics.
  
• Antidiabetic Effects of Camel Milk in Streptozotocin-induced Diabetic Rats
  This study was conducted to analyse the possible antidiabetic effects of camel milk in streptozotocin (STZ)-induced diabetic rats by assaying liver and kidney clinical function parameters. Administration of streptozotocin (55 mg kg ^-1 b.wt.) to the experimental groups of rats resulted in marked detectable changes. The rats were fed daily with fresh camel milk by feeding bottles for 30 days. The effects of camel milk on blood glucose, serum proteins, urea, uric acid, creatinine, lipid profile and the activities of diagnostic marker enzymes of liver function and Alkaline Phosphatase (ALP) were examined in the plasma/serum of control and experimental groups of rats. Camel milk feeding to diabetic rats significantly reduces the levels of blood glucose, urea, uric acid and creatinine and increases the activities of albumin, albumin/globulin ratio and restores all liver function marker enzymes and lipid profile to near control levels. The present study shows that feeding of camel milk to diabetic rats has antihyperglycemic effects and consequently may alleviate liver and renal damage associated with streptozotocin-induced diabetic rats.
  
• Tetranucleotide Repeat Polymorphism in the 3' UTR of the Human Leptin Gene and Risk of Type 2 Diabetes Mellitus in a South Indian Population
  Type 2 Diabetes Mellitus (T2DM), an increasingly common serious metabolic disorder has obesity as one of its major predisposing factors. Accumulating evidence suggests that the adipocyte-derived multifunctional hormone, leptin, may play a vital role in the modulation of obesity and subsequent T2DM. The association of polymorphic tetranucleotide repeat (TTTC) _n polymorphism in the 3' UTR of the human Leptin gene (LEP-tet) with T2DM was examined in the present study. The study population comprised of 481 subjects including 305 patients and 176 controls. Genotyping of the polymorphic variant was carried out by PCR followed by agarose gel electrophoresis. The alleles of the polymorphism consisted of two groups with different size distributions: the shorter group (class I) and the longer group (class II). Analysis of allele frequencies and genotype frequencies between T2DM subjects and healthy controls revealed no significant differences. Risk estimates for different combinations of the LEP-tet genotypes showed that the I/II genotype was associated with modest 1.46 fold increased risk of T2DM (p = 0.04; 95% CI = 1.01-2.12). A similar trend could not be observed after stratification of the subjects based on sex or obesity. The results obtained in the present study indicated an insignificant influence of the leptin tetranucleotide repeat polymorphism on the development of T2DM. Further studies on a similar aspect would help clarify the issue.
  
• Evaluation of Novel Tasar Silkworm Feed for Antheraea mylitta: Its Impact on Rearing, Cocoon Trait and Biomolecular Profile
  The Antheraea mylitta produces tasar silk having massive demand in international market. Its rearing is carried out in outdoor conditions and during course of I to III instar rearing, a major proportion (20-30%) of larvae die due to vagaries of nature, pests and predators etc. which considerably affects the production and productivity. The solution to this major bottleneck of the silkworm rearing lies in evolving a suitable tasar silkworm feed (semi-synthetic diet) for young age tasar silkworm. In present study, comparative evaluation was done among semi-synthetic diet fed; fresh leaf fed indoor reared and fresh leaf fed in natural outdoor reared insects. Data revealed that young age survival and Effective Rate of Rearing (ERR) were higher when larvae were brushed on semi-synthetic diet in contrast to indoor rearing on fresh leaf and complete outdoor rearing. Semi-synthetic diet fed larvae showed greater body weight and their cocoon showed higher weight, shell weight and shell ratio than controls. Concentration of hemolymph protein was slightly higher in semi-synthetic diet fed than outdoor reared larvae whereas, significantly lower in case of indoor reared. Hemolymph protein SDS-PAGE analysis indicates that, the semi-synthetic diet fed larvae is closer to complete outdoor reared larvae than indoor reared. Comparative assessment of rearing, cocoon trait and biomolecular profile of A. mylitta it is assumed that tasar feed developed by our laboratory is novel. It will be helpful in minimizing impact of unfavorable condition during I crop rearing and maintenance of precious eco-races stocks to enhance productivity of tasar silk.
  
• A Novel Oligo-DNA Probe Carrying Non-nucleosidic Silylated Pyrene Derivatives: Synthesis and Excimer Forming Ability
  Sequence-selective DNA detection is an important tool for monitoring many biological processes and for other biotechnological applications. In this way, molecular beacons (MBs) are highly selective in detecting oligonucleotides that have got widespread applications in nucleic acids chemistry. Excimer formation is a fundamental property of pyrene that can be used for designing unique class of DNA probe such as molecular beacons. To develop a new DNA detection probe that gives specific excimer fluorescent signal before hybridization to the target, a novel 29mer stem-loop structured (molecular beacon) DNA was synthesized. Then two non-nucleosidic silylated pyrene units have been incorporated into this novel oligo-DNA by standard phosphoramidite method using automated DNA synthesizer. Molecular beacon probe with silylated pyrene has already been reported by our group where pyrene was used as a part of nucleobase. In the present case, pyrenes are incorporated as non-nucleobase that have successfully displayed excimer fluorescence signal. Interestingly, the resulting modified DNA exhibited strong excimer fluorescent signal both before and after hybridization to the fully matched complementary target. Careful selection of the nucleobases in the stem segment could result in a more efficient sequence discriminating oligo-DNA probe.
  
• Implications of Raised Folate and Lowered Vitamin B12 in Osteosarcoma
  Osteosarcoma is the most common primary tumor of bone, occur mainly during childhood and adolescence. No reports are available in literature where serum folate and vitamin B ₁₂ have been assessed in osteosarcoma patients. Hence, the present study was planned to analyze status of folate and vitamin B ₁₂ in thirty osteosarcoma patients and compare it with thirty controls (subjects with musculoskeletal pain). Serum calcium, alkaline phosphatase levels were higher and phosphorus levels were comparable in osteosarcoma patients as compared to controls. Serum folate levels were significantly raised in osteosarcoma patients as compared to controls (p <0.001). Serum vitamin B ₁₂ levels were significantly lowered in osteosarcoma patients as compared to controls (p <0.001). Folate-mediated one-carbon metabolism (FOCM) is unequivocally linked to multiple health outcomes, including birth defects, several types of cancer and possibly cardiovascular disease and cognitive function. Tumors are known to up regulate FRα modulating the folate uptake in serum. Vitamin B ₁₂ diminishes osteoblastic activity and lowered serum vitamin B ₁₂ levels are possibly due to increased metabolic demand of the tumor. The present study suggests that these parameters can serve as useful markers for diagnosis and follow up of disease.
  

  No Issue Number

• Homology Modeling of a Fruit Ripening Specific Plant MADS–box Factor
  A MADS-box (Minichromosome maintenance-1, Agamous, Deficiens and Serum response factor) transcription factor namely SlMADS RIN ( Solanum lycopersicum MADS Ripening inhibitor) seems to act as global regulator in climacteric fruit ripening process of tomato. Structure modeling of any plant MIKC (MADS-box, I-box, K-box and C-box) -type MADS-box factor were unknown till date, and the present study is an approach towards this direction. The template search of SlMADS RIN was performed by PSI BLAST (Position-Specific Iterative Basic Local Alignment Search Tool). Initial model was built with the help of MODELLER 9v4 package. The predicted 3D structure for SlMADS RIN protein was further validated by Ramachandran plot analysis using the PROCHECK tool. The submitted sequence of SlMADS RIN protein to PSI BLAST tool identified only one region (1-73 amino acids). DOPE (Discrete Optimized Protein Energy) score analysis revealed that the modeled structure showed overall lower DOPE score value (-3968.569336). Ramachandran plot analysis revealed that 94.1% residues were in favored region and 05.9% residues were in allowed region. Result revealed that the SlMADS RIN protein structure under study deviate largely by sequence with the known MADS-box, except the N-terminal 74 amino acid. Further, the side chain and loops of SlMADS RIN showed <1Å (0.233 Å) root mean square deviation. Thus, it can be concluded that this is the first report on prediction of three dimensional models for SlMADS RIN and this modeled structure can be used to predict the molecular function of the protein.
  
• Detection of Canine Parvo Virus by Polymerase Chain Reaction Assay and its Prevalence in Dogs in and Around Mathura, Uttar Pradesh, India
  The early detection of the Canine Parvo Virus (CPV) is of paramount importance. The present study was aimed to know the molecular epidemiology of Canine parvo virus. Canine faecal samples from 100 dogs showing the clinical signs of gastroenteritis in and around Mathura, Uttar Pradesh, India were collected and DNA was extracted by phenol-chloroform method. CPV vaccine strain was used as a positive control. Polymerase Chain Reaction (PCR) was carried out to amplify VPI/VP2 gene using a set of 20-mer primers [pCPV-RT (Forward): 5’-CAT TGG GCT TAC CAC CAT TT-3’; (Reverse): 5’-CCA ACC TCA GCT GGT CTC AT-3’)] from position 3136-3155 to 3276-3295 of VP1/VP2 gene. A PCR product of approximately 160 bp was generated with positive faecal samples and CPV vaccine strain. After screening, 63 dogs were found positive for CPV but no sex variation was noticed amongst the CPV positive cases. Dogs, of the age group of ≤6 months were more susceptible in comparison to of >6 months and highest occurrence was noted in unvaccinated dogs and dogs in co-habitation with other dogs. Breed wise distribution of CPV in dogs revealed that the prevalence of CPV was the highest in Doberman (77.78%), followed by Spitz (78.57%), German shepherd (70.00%), Labrador (68.75%), Pomeranian (45.45%). It is concluded that CPV is prevalent in the Mathura and nearby area and it is more common in pups of age less than 6 months old and more prevalent in German shepherd, Labrador and Pomeranian breeds of dog.
  
• Partial Characterization of Proteolytic and Milk Clotting Enzymes in Sodom Apple Calotropis procera (Ait.) R.Br. (Asclepiadaceae) Plant
  Calotropis procera (Sodom apple) or Bomubomu in Yoruba Language has been used locally in the Northern part of Nigerian and the Fulani set-ups in African settings as curdling agent in the production of ‘warankasi’ (a local soft cheese) when using cow milk; this plant is also known for its medicinal importance like antidote for snake bite, sinus fistula, rheumatism, mumps, burn injuries, body pain and treatment of jaundice in India and some parts of Africa. However, this present study was undertaken to characterize the milk clotting and proteolytic enzymes in the plant with the aim of establishing the optimal conditions of the enzymes to be used for cheese and allied milk products manufacturing and protein hydrolytic processes of medicinal importance . The results obtained from this study showed that the enzyme activities were evidenced in all the plant parts extract preparations except the root that had only proteolytic activity without noticeable milk-clotting activity for the period of one hour of investigation. The optimal pH for proteolytic and milk clotting enzyme activities were observed to be 4.0 for proteolytic and 5.0 for milk clotting enzyme activities in the crude extracts using casein and powdered milk as substrates, respectively while the temperature optima for the two enzyme activities were observed to be 70°C for milk clotting and 60°C for proteolytic. Calcium ion and cysteine were found to activate the two enzymes, with maximum activation at 800 mM Ca ²⁺ for proteolytic activity and 10 mM for milk clotting activity while cysteine showed maximum activation at concentration of 6.0 mM for the two enzymes as Pb ²⁺ ions caused inhibition of both enzymes at the very least concentration. The results of analyzes from this work showed how the proteolytic and milk clotting enzyme activities of the plant latex could be enhanced for protein hydrolytic processes of medicinal importance and industrial production of cheese and allied milk products.
  
• Induction of Salicylic Acid in Cucumber Plants Against Cucumber Mosaic Cucumovirus Using Biotic Inducers
  Systemic Acquired Resistance (SAR) could be induced in cucumber plants using different individual of seven microbial isolates against Cucumber mosaic Cucumovirus (CMV). These isolates were Bacillus subtilis , Bacillus polymyxa , Bacillus circulans , Pseudomonas putida , Pseudomonas fluorescens 2 and Pseudomonas fluorescens 8 as bacterial isolates and Trichoderma harzianum as fungal isolate. The occurrence of SAR was found by seed treatment with microbial liquid culture based on virus infectivity and the level of free and endogenous salicylic acid (SA), 14 days from CMV inoculation. Seven biotic inducers reduced CMV infectivity at range 16.6 to 39% and T. harzianum gave the highest percentage of reduction 39%. In addition, the level of total SA has been increased in treated plants, B. subtilis gave the highest level of SA 239.13 μg g ^-1 fresh weight (fwt) while, B. circulans gave the lowest level 70.1 μg g ^-1 fwt.
  
• Ultrasonication of Chicken Natural Actomyosin: Effect on ATPase Activity, Turbidity and SDS-PAGE Profiles at Different Protein Concentrations
  With the increasing application of ultrasonics in meat tenderization and processing, physicochemical events initiated by sonic radiation at myofibrillar level and propagated in complex tissue such as meat require a clear understanding. The enormous amount of basic information collected by studying myofibrils, actomyosin or their individual constituents has already clarified intricacies of muscular contraction and, part of such basic information has find application in meat sciences. In this investigation, chicken Natural actomyosin (NAM) has been taken as a simple model to work out some effects of ultrasonication in a concentration range of 0.5 to 1.8 mg protein mL ^-1 . At each concentration, NAM solution in 0.6 M NaCl (2.0 mL) was individually exposed to 20 kHz sonic waves for a total of 10 min. Cooling was maintained by keeping NAM containers in crushed ice and a lag of 5 sec after each 10 sec long sonic burst. Aliquots from each sonicated NAM were subjected to biochemical analyses. Most striking differences were observed in Ca ²⁺ -ATPase activity, which displayed a steady decline that corresponded with the decreasing protein concentration. Ultrasonication of NAM for 10 min caused a loss of ~47% of Ca ²⁺ -ATPase activity at the highest dilution (0.5 mg mL ^-1 ). In the same order of protein concentration, turbidity of ultrasonicated NAM also decreased which denotes increasing transparency. Thus, ATPase and turbidity data demonstrate that due to sonic radiation, interactions among constituents of chicken actomyosin complex alter and these structural changes are devoid of any fragmentation. Under present experimental conditions, SDS-PAGE profiles did not reveal any novel band which could be attributed to ultrasonic fragmentation or proteolytic contamination. The findings also suggest that unlike myofibrils, actomyosin is a model where interactions and substructural changes of constituent polypeptides can be investigated without interference of endogenous muscle proteases.
  
• Hypolidemic and Angiotensin-converting Enzyme Inhibitory Effect of Peristrophe bicalyculata (Retz.) Nees
  The objective of this study was to investigate the effect of Peristrophe bicalyculata on lipid levels and activity of angiotensin-converting enzyme in kidneys, lungs and heart of fat-fed obese Wistar rats . The angiotensin-converting enzyme inhibition pattern of Peristrophe bicalyculata was also studied. Apparently healthy male Wistar rats were grouped into: control group, fat-fed control, standard control (atorvastatin at 70 mg kg ^-1 ) and low dose (100 mg kg ^-1 ) and high dose (250 mg kg ^-1 ) extract groups, after confirmation of the lethal dose (LD ₅₀ ). All rats, except those in the control group were fat-fed until obesity was confirmed (BMI>0.3) and treatment given for 4 weeks. From our results, serum and liver levels of Total Cholesterol (TC), Triacyglycerol (TG) and Low Density Lipoprotein increased significantly (p<0.05) in obese rats compared to the control rats; while High Density Lipoprotein (HDL) significantly decreased. Administration of atorvastatin and the extract significantly (p<0.05) reduced TC, TAG and LDL levels and increased HDL in serum and liver. The activity of Angiotensin Converting Enzyme (ACE) in kidneys, lungs and heart of obese rats was significantly (p<0.05) higher than that of rats in the control group. The extract reduced ACE activity in the lungs, kidneys and heart by 97, 98 and 96%, respectively, which was 20, 30 and 15 folds that of the standard drug. Studies on the inhibition pattern of the extract showed that Peristrophe bicalyculata inhibits ACE by competing with the substrate for the active site, exhibiting a competitive type of inhibition. The K _m of ACE increased from 0.25 to 2.5 mM at 1% concentration and 5 mM at 2% with an unchanged V _max and k _i value of 1.1 mg mL ^-1 . In conclusion, this study has confirmed that the aqueous extract of Peristrophe bicalyculata may be useful in the reduction of cardiovascular disease risk.
  
• Modulatory Effect of Aqueous Stem Bark Extract of Psidium guajava Linn. against CCl4 Induced Liver Damage in Rats
  The present study was aimed to evaluate the stem bark aqueous extract of Psidium guajava for modulatory effect against CCl ₄ induced liver damage in rats. A total of thirty six male rats, were randomly divided into six groups of six rats each. The extract was administered orally for 15 days at 125, 250 and 500 mg kg ^-1 b.wt. The results obtained showed that treatment with the extract significantly (p<0.05) restored liver weight. There was significant (p<0.05) increase in the level of Packed Cell Volume (PCV), haemoglobin (Hb) and Red Blood Cell (RBC) counts and significant (p<0.05) decrease in White Blood Cell (WBC) counts compared to toxin control group. Also administration of the extract caused significant (p<0.05) decrease in the activities of Alanine Transaminase (ALT), Aspartate Transaminase (AST), Alkaline Phosphatase (ALP) and the level of total bilirubin and significant (p<0.05) increase in total protein level compared to toxin control group. Similarly the extract caused a significant (p<0.05) increase in the activities of Catalase (CAT) and Superoxide Dismutase (SOD) and significant (p<0.05) decrease in reduced Glutathione (GSH) and Thiobarbituric Reactive Substances (TBARS) level compared to group 2 (toxin control group). The histopathological study indicated that treatment with the extract restored and regenerated hepatic cells compared to toxin control group. This study found that administration of aqueous stem bark extracts ameliorated hepatotoxicity induced by CCl ₄ in rats.
  
• Oxidative Stress Biomarkers and Biochemical Profile in Broilers Chicken Fed Zinc Bacitracin and Ascorbic Acid under Hot Climate
  Dietary manipulation of heat induced oxidative stress is one of the most widely used method to alleviate the negative effect of heat stress. The combination of two or more potential antioxidants are known to protect against heat stress. Therefore, 200 day old broiler chicks were raised under hot climate to determine the effect of ascorbic acid and zinc bacitracin and their combination as antioxidants. Birds were divided into four treatments; the first treatment (T ₁ ), the control was provided with basal diet. Second (T ₂ ) and third (T ₃ ) treatments were provided with 1 g ascorbic acid and 100 mg zinc bacitracin per kg basal diet, respectively. The fourth treatment (T ₄ ) was provided with a combination of T ₂ and T ₃ for 42 days. Performance parameters were reported every fortnight. Blood samples were collected every fortnight and the harvested sera were used to determine oxidative stress biomarkers. Ascorbic acid and zinc bacitracin were safe for heat stressed birds since the liver and kidney function indicators were not changed. Ascorbic acid and zinc bacitracin lowered lipid peroxidation level (Malondialdehyde; MDA) and increased the activity of catalase (CAT), glutathione-S-transferase (GST) and Super Oxide Dismutase (SOD) whereas, glutathione peroxidase (GPx) activity remained unchanged in all treatments. Serum ascorbic acid concentration was significantly (p<0.05) increased accompanied with hypocholesterolemia in heat stressed broilers receiving ascorbic acid compared to the control. Performance parameters were not influenced by feed additives. In conclusion, ascorbic acid was more potent than zinc bacitracin and their combination in increasing the level of enzymatic and non enzymatic antioxidants.