Showing papers in "American Journal of Biochemistry and Biotechnology in 2010"

Production of biodiesel by enzymatic transesterification: review.

Abstract: Problem Statement: The research on the production of biodiesel has increased significantly in recent years because of the need for an alternative fuel which endows with biodegradability, low toxicity and renewability. Plant oils, animal fats, microalgal oils and waste products such as animal rendering, fish processing waste and cooking oils have been employed as feedstocks for biodiesel production. In order to design an economically and environmentally sustainable biodiesel production process, a proper understanding of the factors affecting the process and their relative importance is necessary. Approach: A comprehensive review of the literature on the subject of biodiesel production was carried out. Traditionally biodiesel has been produced using either acid or base catalysts. The multi-step purification of end products, wastewater treatment and energy demand of the conventional process has lead to search for alternative option for production of biodiesel. The use the enzyme lipase as a biocatalyst for the transesterification reaction step in biodiesel production has been extensively investigated. Lipase is produced by all living organisms and can be used intracellularly or extracellularly. Conclusion: To date, the most popular microbes used for their lipases have been filamentous fungi and recombinant bacteria. A summary of lipases used in transesterification and their optimum operating conditions is provided. In addition to the choice of lipase employed, factors which make the transesterification process feasible and ready for commercialization are: enzyme modification, the selection of feedstock and alcohol, use of common solvents, pretreatment of the lipase, alcohol to oil molar ratio, water activity/content and reaction temperature. Optimization of these parameters is necessary in order to reduce the cost of biodiesel production. Use of no/low cost waste materials as feedstocks will have double environmental benefits by reducing the environmental pollution potential of the wastes and producing an environmentally friendly fuel.

The Development of a Revised Canadian Myalgic Encephalomyelitis Chronic Fatigue Syndrome Case Definition

Abstract: Problem statement: Several investigators have indicated that case defi nitions for Chronic Fatigue Syndrome (CFS) are characterized by vaguely worded criteria that lack operational definitions and guidelines. The most widely used CFS case definition is the Fukuda et al. criteria, which uses polythetic criteria (i.e., patients are only requir ed to have four out of a possible eight symptoms). Yet two of these eight symptoms (post-exertional malais e and memory/concentration problems) are an essential feature of this illness and the Fukuda et al. criteria do not require that these symptoms be present among all patients. Significant methodologi cal problems could occur if investigators in different settings recruit samples with different p ercentages of these core symptoms. In contrast, the Canadian clinical case definition does require spec ific ME/CFS symptoms such as post-exertional malaise and memory/concentration problems. The provision of operationally explicit, objective criteria on specific key symptoms might reduce criterion var iance as a source of unreliability. In addition, th e use of structured interview schedules will ensure t hat symptoms are assessed in a consistent way across settings. Conclusion/Recommendations: In this article, we specified explicit rules for determining whether critical symptoms meet ME/CFS criteria using a revised Canadian case definition and a questionnaire has been developed to assess co re symptoms. It is hoped that these developments will lead to increased reliability of this revised Canadian case definition as well as more frequent u se of these criteria by investigators.

A Proteomics Analysis of Drought Stress-Responsive Proteins as Biomarker for Drought-Tolerant Sugarcane Cultivars

Abstract: Problem statement: The prime objective in breeding selection process of drought-tolerant sugarcane is to identify the correlating marker, wh ich could lead to rapid screening for drought-toler ant cultivars. In this study, we have reported an unkno wn 18-kDa protein (p18) along with other stress-ind ucible proteins to be highly expressed in sugarcane leaves under drought stress condition. Approach: The 2D- PAGE patterns of proteins were compared between those expressed in drought-tolerance K86-161 and drought-susceptible Khon Kaen 1 cultivars. The inte rested proteins were identified by mass spectrometr y. The correlation between p18 expression and drought tolerance was verified in additional 4 sugarcane cultivars using ELISA and western blotting. Two physiological indexes, Chlorophyll content and SOD activity were also evaluated. Results: Mass spectrometry and comparison with known sequences in the database reveal that the proteins expressed only in stressed K86-161 are serine protease inhibitor and the one similar to replication protein A1. A group of p roteins up-regulated in K86-161 are S- Adenosylmethionine decarboxylase proenzyme (SAM), ubiquitin and p18. From ELISA and western blotting analysis, we found that p18 expressed in d rought-tolerant sugarcane cultivars had higher bind ing specificity to antibody than that in drought-suscep tible sugarcane cultivars. Two physiological indexe s showed higher levels in drought-tolerant than those in drought susceptible sugarcanes. Conclusion: These high levels of chlorophyll and SOD are in agreement with a high level of p18 expression in drought- tolerant sugarcanes. It is likely that an accumulat ion of p18 is a response to water deficit. In concl usion, p18 might be a good candidate for development as a marker in drought-tolerant plants.

Effect of Plant Growth Regulators on Callus, Cell Suspension and Cell Line Selection for Flavonoid Production from Pegaga (centella asiatica L. urban)

Abstract: Problem statement: Considering pegaga medicinal properties and over-exploitation, the requirement for a tissue culture technique as an alternative production system was crucial. Approach: Investigation of cell suspension culture response to different plant growth regulators (PRGs) for flavonoid production from elite cell line was carried out. Callus cultures were initiated from the leaf explants of Centella asiatica on Murashige and Skoog (MS) medium containing B5 vitamins and 30 g L−1 sucrose supplemented with different concentrations (0.5-2.5 mg L−1) of 2,4-D, NAA, Dicamba, Picloram and IBA supplied singly and in combination with different concentrations (0.5-1.5 mg L−1) of kinetin, BAP and TDZ. Results: Callus induction was observed for all the PGRs tested. The highest callus induction frequency (86.67%) was observed in MS medium containing 2.0 mg L−1 2,4-D while the combination of 2.0 mg L−1 2,4-D and 1 mg L−1 kinetin in MS medium gave the highest biomass yield (0.27 g dry weight culture−1). This combination was also found to be best for callus proliferation for all the accessions investigated. Among the four accessions tested, UPM03 was found to have the highest biomass yield (0.041 g DW culture−1) and hydrolysed flavonoid content (10.75 mg g−1 DW) after the 12th day of culture. The flavonoids present in the four accessions were quercetin, kaempherol, luteolin and rutin based on high performance liquid chromatography (HPLC) analysis. These results indicated that C. asiatica accession UPM03 was the potential elite cell line in mass production of flavonoid, especially luteolin. Coclusions/Recommendations: In the establishment of cell suspension culture, 2 mg L−1 2,4-D and 1 mg L−1 kinetin were the best PGRs in supporting the cell growth and flavonoid production. This is the first report on the use of PRGs on the establishment of cell suspension cultures in flavonoid production of C. asiatica.

Effect of Camel's Milk on Cisplatin-Induced Nephrotoxicity in Swiss Albino Mice

Abstract: Problem statement: Nephrotoxicity is a major complication and a dose limiting factor for cisplatin therapy. Cisplatin mediated nephrotoxicity is remarkably documented by reactive oxygen species. Camel's milk has good nutritive value, antigenotoxic and anticytotoxic effects. The aim of the present study was to assess the protective effect of camel's milk against Cisplatin-induced renal oxidative stress in mice. Approach: Forty mal Swiss albino mice were randomly divided into four groups (n = 10). Group I, control group. Group II was received cisplatin (12 mg kg-1) for 5 alternate days. Group III was received camel's milk (33 mL kg-1) for consecutive 30 days. Group IV was received camel's milk (33 mL kg-1) for consecutive 30 days before administration of Cisplatin. Results: Cisplatin-induced oxidative stress was indicated by increased level of tissue Malondialdehyde (MDA), serum creatinine and urea, decreased the concentration of reduced Glutathione (GSH), Vitamin C (Vit. C) and Vitamin E (Vit. E) and decreased both activities and gene expression of Superoxid Dismutase (SOD), Catalase (CAT), Glutathione Raductase (GR) and Glutathione Peroxidase (GPx). Camel's milk reduced these biochemical changes and counteracted the deleterious effects of cisplatin Conclusion: The present study demonstrated the renoprotective potential of camel's milk against cisplatin-induced oxidative stress and renal dysfunction in mice. Hence, camel's milk has a potential to be used as therapeutic adjuvant in cisplatin nephrotoxicity.

Extraction and Purification of Collagenase Enzymes: A Critical Review

Abstract: Problem statement: Enzymes have vital roles in several industrial processes (foods, cosmetics, nutraceuticals and pharmaceuticals) due to their highly selective nature and high activity at very low concentrations. Recent efforts to identify new sources of useful enzymes have been concentrated on the marine environment because of the potential to make use of processing wastes. About 35-50% of the mass of the fish caught is a waste that is disposed off at sea or in landfills. The extraction of enzymes from fish processing waste can reduce environment problems and improve the economics of the fish industry. Collagenases are a group of enzymes that can be extracted from fish waste. Approach: Comprehensive reviews of the literature on the extraction, purification, characterization and use of collagenases was carried out. Results: Collagenases have different molecular weights based on their types and sources. They have the ability to break down the peptide bonds in collagen at physiological pH. They are classified into two types: serine and metallocollagenase. Collagenolytic activities have been shown at a wide range of temperatures (20-40°C) and pH (6-8). Many activators can be used to achive collagenase activity including 4-Aminophenylmercuric Acetate (APMA), trypsin, potassium or sodium thiocyanate, iodoacetamide and potassium iodide. Dithiothreitol (DTT), mercaptoethanol, ethylendiaminetetracetic acid, ophenanthroline and cysteine inactivate the enzyme. Collagenases enzymes can be extracted with a variety of techniques using different buffering systems (tris-HCl, sodium bicarbonate, calcium chloride and cacodylate). All techniques involve the use of ammonium sulphate fractionation and centrifugation to precipitate the enzyme. Collagenases are normally purified using chromatographic techniques such as gel-filtration, ion-exchange and affinity column chromatography. Collagenase can be assayed with a number of methods, including: colorimetric absorbance, viscometry, radioactivity and fluorescence spectroscopy. Collagenases are partly responsible for toughness in red meats and are used as tenderizers in food industry, have application in the fur and hide tanning to ensure uniform dying of leather, used in medicine to treat burns and ulcers, eliminate scar tissues, transplantation of organs. Conclusion: Understanding of the nature of the enzymes and identifying the most suitable resources and the methods for their extraction and purification will have significant impact on the fish processing, food and medical industries.

Recovery of Lead(II) from Aqueous Solutions by Zea mays Tassel Biosorption

Abstract: Problem statement: Major adsorbent materials used in heavy metal ion removal from polluted aqueous streams are expensive and difficult to regenerate. In this study, the possibility of using Zea mays tassel, as an alternative low cost biosorbent material to remediate heavy metal pollution was investigated. Lead (II) was used because of its wide application in industrial products and well documented toxicity. Approach: Tassel was obtained from mature Zea mays cultivar R52 hybrid plants. The tassel was milled to a powder and was used to adsorb lead(II) ions from simulated solutions in batch experiments. The desorption of lead(II) was carried out using nitric acid and sodium citrate solutions. The adsorbent was characterized by FTIR, EDX and ESCA before and after application of lead(II) solutions. Results: For samples with concentrations of 100 mg L-1 Pb(II), 94-98% was adsorbed and 57-74 and 57-67% recoveries were achieved with 0.5-5 M nitric acid and 0.01-0.2 M sodium citrate as the stripping solutions, respectively. EDX spectrum of pure tassel indicated that group 1 and 2 metals were the major exchangeable ions present on its surface. ESCA analysis picked up small amounts of lead(II) in the form of Pb(OH)+ and Pb(NH)+ ions on the surface of tassel adsorbent exposed to Pb2+ ions and none on pure tassel sample. Functional groups such as -H, -NH2, -C = O and -COOH which are polar and are legends which are capable of binding heavy metals were identified by FTIR. Conclusion: The potential of Zea mays tassel to adsorb and recover heavy metals from aqueous solution was successfully demonstrated with Pb(II) sample solutions. The results obtained thus far demonstrated the possibility of using tassel powder in the removal as well as recovery of metals from aqueous solutions.

Antilithiatic Influence of Spirulina on Ethylene Glycol-Induced Nephrolithiasis in Male Rats

Abstract: Problem statement: Nephrolithiasis or renal stone disease remains a significant health problem in the adult population. Nephrolithiasis is a recurrent disorder prominent in males. It is significant medical and surgical problem because of incidence, recurrence and severe consequences. The present day medical management of nephrolithiasis is either costly or not without side effects. Invasive procedures for the treatment of nephrolithiasis may cause serious complications and they also impose a great load of costs to the healthcare system. Hence the search for antilithiatic drugs from natural sources has assumed greater importance. Approach: The aim of the present study was to evaluate the antilithiatic activity of spirulina supplementation on ethylene glycol induced nephrolithiasis in male rats. Healthy male Wistar rats were used in the present study and were divided randomly into 4 groups. Rats of 1st group were served as normal control. Rats of 2nd group were received 0.75% ethylene glycol in drinking water for three weeks and drinking tap water for the next three weeks. Rats of 3rd group were received 0.75% ethylene glycol in drinking water for three weeks and fed with spirulina solution (20 mg kg-1 body weight) for the next three weeks. Animals of 4th group were treated as 1st group for the first three weeks and fed with spirulina solution ate the same dose given to 3rd group for the last three weeks. After six weeks, serum levels of sodium, chloride, potassium, calcium, phosphorus, Blood Urea Nitrogen (BUN), uric acid, creatinine, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST) were measured. Results: Statistically increases in the levels of sodium, chloride, BUN and ALT and a decrease in the level of calcium were noted in rats treated with ethylene glycol. Supplementation of spirulina for the last three weeks mostly recovered the rats from nephrolithiasis and completely from hepatotoxicity induced by ethylene glycol. Conclusion: This study suggested that spirulina is a safety and promising agent as a functional food for the management of nephrolithiasis induced by ethylene glycol and may be also by other chemical factors.

The effect of Citrullus colocynthis pulp extract on the liver of diabetic rats a light and scanning electron microscopic study.

Abstract: Problem statement: The goal of the current investigation was to clarify the effects of Citrullus colocynthis pulp extract on the structure of the liver of diabetic rats at both light and scanning electron microscopic levels. Approach: Forty-eight adult male albino rats were equally allocated into four groups: Group1: control, Group 2: Citrullus colocynthis-treated, Group 3: diabetic rats and Group4: diabetic rats treated with Citrullus colocynthis. All treatments were administered via an intragastric tube. Diabetes was induced in the rats of groups 3 and 4 by an intraperitoneal injection with alloxan. Results: The liver of Citrullus colocynthis-treated rats revealed minor histological changes versus the control animals. In group 3 animals, diabetes caused degenerative alterations in the form of disorganization of the hepatic cords, cytoplasmic vacuolization and pyknosis of the nuclei of hepatocytes and inflammatory cell infiltration. Scanning electron microscope examination of these livers revealed numerous lipid droplets within hepatocytes, damaged blood sinusoids and hemorrhage of erythrocytes between hepatocytes and inside Disse's spaces. On the other hand, the normal histological and scanning ultrastructural features were nearly resumed in the liver of diabetic rats treated with Citrullus colocynthis pulp extract. Conclusion: The present study proved a lessening effect of Citrullus colocynthis pulp extract on the liver of diabetic rats. In light of these advantageous influences, it is advisable to widen the scale of its use in a trial to alleviate the diabetic hepatic adverse effects.

Apoptosis of human Breast Cancer Cells induced by Ethylacetate Extracts of Propolis

Abstract: Problem statement: Propolis has been ethno medically claimed to possess a wide array of biological activities including anticancer activity. The purpose of this research was to verify the folklore claim. Approach: This study was performed in a human breast carcinoma cell, MCF-7. Extract of propolis from different solvent, ethylacetate and n-buthanol showed induced apoptotic cells was detected by flow cytometry. Results: The results demonstrated that ethylacetate extract of propolis can induce apoptosis in MCF-7 as large as 13.21% during the 24 h incubation. On the other hand, doxorubicin is able to induce apoptosis as large as 18.89% during the 24 h incubation. Conclusion: The extracts of propolis ethylacetate had cytotoxic activity and triggers apoptosis on MCF-7 cells.

Effect of Exercise Training on Adiponectin Receptor Expression and Insulin Resistance in Mice Fed a High Fat Diet

Abstract: Problem statement: Adiponectin is an adipocyte secreted hormones, exerts its effects via the specific receptors. AdipoR1 and adipoR2 and plays a pivotal role in lipid and glucose metabolism. Approach: We investigated the effect of increased physical activities on insulin resistance and if this effect is modulated through adiponectin receptor expression. Results: We also assessed the effect of High Fat Diet (HFD) on adipoRs expression. Mice were subjected to 16 weeks of HFD protocol then to 4, 6 and 8 weeks of exercise training. Following the experimental protocol the fasting plasma glucose, insulin and index of Homeostasis Model Assessment of insulin Resistance (HOMA-R) were evaluated. The mRNA expression of adiponectin receptors genes were also analyzed using reverse transcription RT-PCR. The consumption of high fat diet by the mice for 16 weeks resulted in a significant increase in weight associated with insulin resistance and associated with significant decrease in adiponectin receptors R1 and R2 expression in both liver and skeletal muscle. Exercise training for 4 weeks resulted in a significant improvement in the insulin resistance state, significant increase in expression of AdipoR1 and AdipoR2 in both liver and skeletal muscles. On increasing duration of exercise training for 6 weeks and 8 weeks there was significant improvement in insulin resistance and significant increase in the expression of AdipoR1 in liver, AdipoR1 and AdipoR2 in muscle, whereas liver AdipoR2 expression was significantly decreased. Conclusion/Recommendations: we conclude that the exercise training improves insulin sensitivity and up-regulates mRNA expression of AdipoR1 in both skeletal muscle and liver in mice and AdiopR2 in muscle and this suggests that the insulin sensitizing effect of exercise training may be mediated even partially through increased adiponectin receptor expression and up regulation of adiponectin receptors reaches certain level and increasing the duration of exercise adds no more improvement to insulin sensitivity or adiponectin receptor expression.

Modulation of Insulin Receptor Substrate-1 and Some Inflammatory Variables in Hyperinsulinemic Rats Treated with Cinnamon Extract

Abstract: Cinnamon Extract (CE) has shown to be generally safe when ingested and to have many pharmacological properties. Problem statement: Study the effects of daily intake of CE on the modulation of hepatic, cardiac Insulin Receptor Sub strate-1 (IRS-1) and their relations to some inflammatory variables in hyperinsulinemic rats. Approach: The influence of CE administered orally was studied in hyperinsulinemic rats. Eighteen male Wistar rats were divided into 3 groups of 6 rats each. Group 1; control animals received starch as c ontrol diet, while group 2; rats were fed a "high- fructose diet" (60%). Group 3; fructose-fed rats re ceived orally CE (0.5 mL/rat/day) from the 16th day of fructose feeding in experimental period. The ani mals were maintained in their respective groups for 30 days. At the end of the experimental period, ser um levels of glucose, insulin, lipid profile, Total Antioxidant Capacity (TAC), Malondialdehyde (MDA), sialic acid and soluble Fas (sFas) were assayed. Hepatic and cardiac IRS-1 levels were also evaluated. Results: Fed high fructose diet to rats induced significant elevations in serum levels of g lucose, insulin, triacylglycerol, HDL-c, sialic aci d, sFas and MDA, while the level of serum TAC was significantly reduced as compared to controls. Also significant reduction in the levels of hepatic and cardiac IRS-1 were recorded as compared to controls. Oral administration of cinnamon extract to fructose -fed rats alleviated the effects of fructose and th ese rats showed a normal level of the parameters studie d. The percentage changes of IRS-1 level in fructose-fed rats before and after treatment with C E were 38.51 for liver and 31.92% for cardiac muscle. This increase in IRS-1 level after treatmen t is still lowered than control level with the percentage change -11.82 and -9.93% for liver and c ardiac muscle respectively. There was a significant positive correlation between IRS-1 and TAC level whereas there was negative correlation between IRS-1 and MDA, sialic acid and sFas. Conclusion: This study reports interest findings that cinnamon extract enhances glucose uptake by activating insul in receptor kinase activity in rats fed high fructo se diet and it has additional roles as antioxidant and anti-inflammatory agents. Thus, clinical applicati on of cinnamon could be considered as a potential ther apeutic option in humans in the treatment of insuli n resistance states. So inclusion of cinnamon in the daily diet could be recommended.

Modeling growth of microalgae Dunaliella salina under different nutritional conditions.

Abstract: Problem statement: The aim of this study was to find the empirical model that describes the growth kinetics of Dunaliella salina, with low production cost and to estimate parameters of this model. Approach: In this study the strain of D. salina UTEX 200 was cultivated in seawater (0.5 M NaCl) at room temperature with agitation of 150 rpm and luminous intensity of 60 mmols.m-2.s-1. The synthetic medium AS100 (0.2 M NaCl) was used in this study for comparison purposes and in order to determine the optimal growth of the microalgae. Kinetics of growth and β-carotene production was determined in a period of 15 days. Results: After the analysis of the behavior graphic, an unstructured model was used for describing the cell growth (logistic model). It was observed that the model was well adjusted to experimental data for the two conditions of analysis. It was observed that alga produces carotenoids under conditions of stress, in which the cell division are retarded. In the case of cells grown in seawater (higher salt concentration), the cell growth was lower but the concentration of β-carotene was higher. Conclusion: In general, these results suggest that D. salina presents higher potential for β-carotene accumulation and that high salinity decreases cellular concentration (measured by the parameter Xm of the kinetic model proposed); however there is an increase in β-carotene production.

Cloning of the Endoglucanase Gene from a Bacillus amyloliquefaciens PSM 3.1 in Escherichia coli Revealed Catalytic Triad Residues Thr-His-Glu

Abstract: Problem statement: An Indonesian marine bacterial isolate, Bacillus amyloliquefaciens PSM 3.1 was isolated for hydrolyzing cellulose. A 1500-bp nucleotide fragment was amplified from the chromosomal DNA by the use of primers directed against the conserved sequence of Bacilli endoglucanase genes obtained from GenBank. Approach: The fragment was cloned and expressed in Escherichia coli. Results: The endoglucanase gene (eglII gene) had an open reading frame of 1500 nucleotides encoding a protein of 499 amino acids. The EglII protein belonged to Glycosyl Hydrolase family 5 (GH5) with a Cellulose Binding Module 3 (CBM 3). The structure model of the EglII protein revealed that the catalytic residues seemed to be Glu169 (as proton donor) and Glu257 (as nucleophile) and the catalytic triad residues were Thr256, His229 and Glu169. The EglII endoglucanase exhibited an optimum pH of 6.0 and temperature of 50°C and the enzyme tolerated to high salt concentration. Conclusion/Recommendations: This EglII endoglucanase is a promising candidate for many applications in biomass degradation.

Expression of Adiponectin Receptors in Human Placenta and Its Possible Implication in Gestational Diabetes

Abstract: Problem statement: Similar to obese patients and type 2 diabetic patie nts, adiponectin levels are reduced in former Gestational Diabetes M ellitus (GDM) patients and are lower in GDM women during late pregnancy compared with pregnant control subjects matched for BMI. Diabetic insult at later stages in gestation, such as may oc cur in gestational diabetes, will foremost lead to short- term changes in a variety of molecules for key func tions including gene expression in the placenta. Approach: In this study we assessed the expression of adipon ectin receptors in human placenta to identify the site (s) of expression and to clarify the effect of gestational diabetes in this expressi on. This study was carried on 10 normoglycemic pregnant women and 20 GDM women. The placental tissue was collected immediately after delivery and tissue biopsies were taken from both fetal and maternal sides of each placenta. One step-RT-PCR for ADIPOR1 and ADIPOR2 was done by Real Time PCR using Syber Green technique. Relative quantification of mRNA of the ADIPOR1 and ADIPOR2 genes was measured using ABI7900 Real Time machine. Results: Both types of Adiponectin Receptors (ADIPOR1 and ADIPOR2) are expressed in human placenta. ADIPOR1 is more highly expressed than ADIPOR2 in both fetal and maternal sides of GDM cases and normal pregnant women. ADIPOR1 mRNA expression was significantly up regulated in GDM women compared to normal pregnant women, whereas no significant difference in the expression of ADIPOR2 was detected between the two groups. There was no evidence of maternal-fetal side difference in the expression of adiponectin receptors in GDM cases but in normal pregnant women there is a statistically significant difference between both s ides in the expression of both ADIPOR1 and ADIPOR2. Conclusion: We concluded that adiponectin plays an important r ole in mediation the glucose metabolism in fetal tissues through its rec eptors, mainly Adiponectin Receptor 1 (ADIPOR1).

Secondary Metabolites in Essential Oil of Achillea millefolium (L.) Growing Wild in East Part of Kosova

Abstract: Problem statement: Chemical composition of essential oil of Achillea millefolium (L.), originated from east part of Kosova, was investigated. Approach: The chemical profile of the essential oil was evaluated by GC and GC-MS. Results: Analysis of the oil resulted in the identification of 33 peaks, representing 81.4% of the oil. The main compounds of Achillea millefolium (L.) from east region of Kosova, were 1,8-cineole (22%), camphor (21%), borneol (7,6%) and β-pinene (5.3%). Conclusion: After comparison of our date with those reported in literature we can conclude that various chemotypes of Achillea millefolium (L.) occur in nature.

Identification a Novel Raw-Starch-Degrading-α-Amylase from a Tropical Marine Bacterium

Abstract: Problem statement: Bacteria from the surface of the tropical marine hard coral Acropora sp. were screened for producing raw-starch-degrading-α-amylase. Approach: Based on its 16s rDNA sequence, a bacterium that produced the highest amylolitic activity was identified as Bacillus amyloliquifaciens ABBD. The bacterial isolate secreted a α-amylase extracellularly and then the enzyme was partially purified by ammonium sulfate precipitation followed by anion exchange chromatography. Results: Electrophoresis results both SDS-PAGE and native PAGE suggested that the enzyme was a heterodimeric protein (97 kDa) consisting of 45 and 55 kDa subunits. The α-amylase had an optimum pH of 7.0 and temperature of 60°C. More than 80% activity of the enzyme was retained under high salt conditions (up to 20% NaCl). The enzyme remained stable at 50°C for 1 h. Starch hydrolysis by the enzyme at 70°C yielded oligosaccharides (G2-G4) and at room temperature yielded glucose/maltose (G1 and G2). Conclusion: The B. amyloliquifaciens ABBD α-amylase was capable of degrading various raw starch granules from corn, rice, cassava and sago at room temperature.

2-Nonenal-Ovulatory Specific Volatiles in Human Saliva throughout Menstrual Cycle by Gas Chromatography and Mass Spectrometry Analysis

Abstract: Problem statement: The present investigation carry out a pilot study of a novel method to identify the salivary volatiles in different phases of menstrual cycle for the assessment of ovulation detection using gas chromatography and mass spectrophotometer. Approach: The profiles from follicular phase (6-12 days); ovulatory phase (13-14 days) and luteal phase (15-26 days) of menstrual cycle samples were compared to establish any qualitative and quantitative differences that might have potential value in human olfactory communication. Dichloromethane was used as the solvent for extraction of the compounds. Results: Fifteen compounds were identified. They include organic compounds like, acid, aldehyde, amine and alcohol. The most important constituent was 2-nonenal, which usually comprised 75% or more of the total volatiles observed in ovulatory phase. The concentration of many constituents varied widely. This appeared to be periodically in three cycles for five of the constituents, with a period of a few weeks and with pronounced maxima at the peak of ovulatory period of which only two were common to all the chromatograms. The chemical profile of ovulatory phase saliva was distinguished by the presence of two specific compounds, viz. 2- Nonenal, Acetic acid and Acetaldehyde that were not found in the other reproductive phases of saliva sample in women. Apparently these compounds are 2-nonenal, dodecanol, acetic acid and acetaldehyde. One or more of these compounds may have pheromonal activity in human body odor. Conclusion: Differentiation of the volatile patterns among reproductive phases in women may help to find the diagnostic marker for ovulation detection.

Antimicrobial Properties of Newly Synthesized Derivatives of Coumarine

Abstract: Problem statement: Coumarins are well known for their biological activity On the basis of that we have synthesized some new derivatives of coumarine and investigated their antimicrobial properties Approach: 4-Heteroaryl-coumarin-3-carbaldehydes 4(a-d) are synthesized by condensation of 4-chloro-coumarin-3-carbaldehydes 2 and corresponding heterorylamines 3(a-d) under reflux reaction conditions Antimicrobial properties of new coumarins 4(a-d) are investigated and results are submitted for their activities against Staphylococcus aureus, Escherichia coli, Hafnia alvei, Pseudomonas aeruginosa and Enterobacter cloacae Applying the Agar disc diffusion technique we measured diameters of the inhibition zone around discs which are previously wetted with N, N-DMF solution of compounds, 1, 3 and 5 mg L −1 Results: The inhibition zone depends from concentrations and also from sort of bacteria The inhibition zone differ from 0 to 30mm Two sort of bacteria, Hafnia alvei and Pseudomonas aeruginosa, are resistant to these new synthesized compounds Conclusion: From results we may conclude that these derivates showed moderate to high activity against Staphylococcus aureus, Escherichia coli and Enterobacter cloaco Compounds 4(a-d) are more active against Staphylococcus aureus, Ecoli and Enterobacter cloaco Compounds 4(a-d) are not active against Hafnia alvei and Pseudomonas aeruginosa

Endothelial Dysfunction Induced by Type 2 Diabetes Mellitus and Fibrinolytic Activity

Abstract: Problem statement: Type 2 diabetes mellitus is the world's largest sy stemic and metabolic disease. Endothelin (ET) was first described as a p eptidergic endothelium-derived constricting factor. Endothelin-1 is one of the endothelial fact ors that is overexpressed in cases of endothelial dysfunction. The fibrinolytic system is present in plasma with the degradation of fibrin polymers in blood clots. It is a proteolytic mechanism that res ults in the formation of one main enzyme, plasmin, which cleaves fibrin. Approach: A total of 20 normal white albino rats weighing be tween 180-200 g were used and divided into two groups each of 10 ra ts, one group served as a control which fed on normal chow diet, while the second group fed on hig h fat diet for one month, then received streptozocin (25 mg kg -1 ) intraperitoneally as a single dose and continued feeding on high fat diet. Plasma was separated to determine the levels of tis sue Plasminogen Activator (t-PA), Plasminogen Activator Inhibitor-1 (PAI-1), Fibrin Degradation P roducts (FDPs) and Euglobulin Clot Lysis Time (ECLT). Results: Data showed that diabetic rats have inhibited fibr inolytic activity as detected from the significant increase in PAI-1, significant decr ease in both t-PA and TDPs with significant prolongation of the ECLT relative to the control gr oup. Conclusion: Endothelial dysfunction is associated with a prothrombotic state.

The inhibition of human pathogens: Trichophyton rubrum and Trichoderma harzianum by a natural product

Abstract: Problem statement: A number of studies have recently conducted to identify novel and potent antifungal components from natural products. One of the reasons is to overcome the antifungal resistant developed against most of commercially available drugs. Studies confirmed that mung beans have increased phenolic compounds and enhanced defenses during germination. Approach: We hypothesized that antifungal activities might be found in sprouts of mung beans, or Vigna radiate (L.) R. Wilczek. The screening method was conducted using disc diffusion assay against 12 fungi. It was followed by the evaluation of the minimum inhibitory concentration and the minimum fungicidal concentration. Results: The screening results revealed a potential antifungal activity by mung bean sprout extract against 2 out of 12 fungi including remarkable antifungal activity against human fungal pathogens, Trichophyton rubrum and Trichoderma harzianum. The potential antifungal activity of mung bean sprout reflects effective quality/quantity of polyphenolic compounds present after bean germination. Conclusion/Recommendations: This unprecedented study showed that mung bean sprout extract is a potential source for novel antifungal compound (s) that is inexpensive and readily available at a large scale for pharmaceutical companies.

Do Resistin and Tumor Necrosis Factor-α Relate to Changes in Insulin Resistance in Normal Pregnancy?

Abstract: Problem statement: The purpose of this study was to evaluate the role of resistin and Tumor Necrosis Factor-α (TNF-α) in insulin resistance during pregnancy. Approach: Serum resistin and TNF-α concentrations were measured by ELISA in 86 healthy pregnant women (26, 23 and 37 of them in the 1st, 2nd and 3rd trimesters, respectively) and in 21 healthy non pregnant women in a cross sectional study. Results: Resistin concentration was significantly higher in the third trimester (9.5±3.3 ng mL-1) as compared with non pregnant women (7±3.3 ng mL-1). Serum TNF-α level were also significantly increase in pregnant women (2.6±1.9 pg mL-1) as compared with maternal healthy controls (0.8 ±0.7 pg mL-1). There were significant correlation between gestational age and BMI (r = 0.28, p = 0.01), resistin (r = 0.36, p = 0.002) and TNF-α (r = -0.44, p<0.0001). There was not significant correlation between gestational age and Insulin Resistance (IR). We also did not found correlation between IR and resistin as well as between IR and TNF-α in pregnant women. Conclusion: TNF-α and resistin do not appear to contribute greatly to pregnancy induced insulin resistance in healthy pregnancy.

Impact of L-Carnitine and Cinnamon on Insulin-Like Growth Factor-1 and Inducible Nitric Oxide Synthase Gene Expression in Heart and Brain of Insulin Resistant Rats

Abstract: Problem statement: Evaluate the effects of daily administration of L- carnitine and cinnamon extract for two weeks on the expression of Insulin-like Growth Factor-1 (IGF-1) and inducible Nitric Oxide Synthase (iNOS) genes in car diac and brain tissues of rats with Insulin Resistance (IR). Approach: Rats were divided into 4 groups (8 animals each): Group (1) rats fed control diet (60% starch) as control while groups ( 2, 3 and 4) fed high fructose diet (60% fructose). At the beginning of the 3rd week of feeding, rats of g roup (3) were treated with L-carnitine (300 mg kg -1 body weight/day, i.p.) and animals of group (4) rec eived a daily oral dose of cinnamon aqueous extract (0.5 mL rat -1 ). The animals were maintained in their respect ive groups for 4 weeks. Results: Feeding high fructose diet causes significant reduc tion in Insulin Receptor Substrate-1 (IRS-1) (amounted 30.65%) and elevation in iNOS expression (reached 51%) in the cardiac tissues as compared to control. In brain tissues, the IGF-1 mR NA was reduced in fructose loaded groups (28.81%). Administration of either L-carnitine or c innamon extract significantly improves the expression of the cardiac studied genes but with no effects on the brain tissues . Conclusion: The present study illustrated that CE was more potent t han L-carnitine in improving the IR. Key word: Insulin resistance, insulin-like growth factor-1, i nducible nitric oxide synthase, insulin receptor substrate-1

Amelioration of Inducible Nitric Oxide Synthase, Insulin like Growth Factor-1 Gene Expression and Insulin Receptor Substrate-1 in Liver Tissue of Insulin Resistant Rats Treated With L-Carnitine

Abstract: Problem statement: It has been reported that genes expression (inducible Nitric Oxide (iNOS). Insulin-like Growth Factor-1(IGF-1) have a role in both glucose homeostasis and insulin resistance. The aim of this study was designed to evaluate the amelioration of the iNOS, IGF-1 genes expression as well as IRS-1 in liver tissues of rats fed high fructose diet treated with Lcarnitine. Approach: About 24 male Wister rats of body weight 120-160 g were divided into 3 groups of 8 rats each. Group 1 received control diet, while group 2 and 3, rats received high fructose diet (60 g 100 g-1 diet). Group 3, after 2 weeks from fructose feeding animals were treated with L-carnitine (CA) (300 mg kg-1 body weight day-1 i. p). At the end of the experimental period (30 days), serum levels of glucose, insulin, Triacylglycerol (TG) and cholesterol were determined. Hepatic contents of cholesterol, triacylglycerol, Malondialdehyde (MDA) and nitrogen oxide products were assayed. Genes expressions of iNOS, IGF-1 as well as IRS-1 were also determined in liver tissues of the experimental animals feeding high fructose diet. Results: Compared to control rats, the high fructose feeding in animals induces alterations in serum glucose, lipid metabolism and hepatic TG and MDA. In addition, fructose fed group develop marked increase in hepatic gene expression of iNOS and pronounced decreases in both IGF-1 mRNA and IRS-1 receptor. The administration of L-carnitine to rats fed high fructose diet mitigated the adverse effects of fructose load (insulin resistance) through the regulation of studied genes expression as well as insulin receptor substrate-1. Conclusion: The important findings of this context indicate the close association between hepatic gene expression (iNOS and IGF-1), IRS-1 receptor and insulin resistance. The exogenous CA to fructose fed rats improves the inflammation resulting from insulin resistance through the amelioration of the studied genes expression. This indicates that iNOS and IGF-1 have the characteristics to be marker of the metabolic syndrome.

Anti β 2 -Glycoprotein I Antibodies in Women with Recurrent Spontaneous Abortion

Abstract: Problem statement: Antiphospholipid antibodies (aPL) are associated with repeated miscarriages and pregnancy complications, however their pathogenic mechanisms are still matter of research. Anti-β2 glycoprotein 1 (β2GPI) antibody is one of these aPL that its contribution to RSA risk remains poorly understood. There were not any data about role this auto-antibody in RSA in South of Iran. This study was conducted to assess Anti- β2GPI IgM and IgG as RSA risk factors for RSA in this area of Iran. Approach: This case-control study was carried out in gynecology special hospital of Hormozgan University of medical sciences, during 2004-2005. A number of 250 women with >3 consecutive idiopathic pregnancy losses as a case group and 200 age matched women with normal full term delivery and negative history of miscarriage as a controls were studied. All obtained sera from the case and the control groups were tested using an Enzyme Linked Immunosorbent Assay (ELISA) method for detection of anti-β2GPI IgM and IgG. Data was analyzed, using SAS 8 statistical software (chi square and t-test). Results: There were significant differences between the prevalence of positive anti-β2GPI IgM (p = 0.01) and IgG (p = 0.02) in the case group and the control group. A positive significant relationship was observed in the case group between number of abortion and seropositivity for anti-β2GPI IgG (p = 0.028, r = +0.181) and anti-β2GPI IgM (p = 0.0381, r = +0.014). Conclusion: This study showed anti-β2GPI antibodies are important causative agent for RSA in this area of Iran.

Total Level of Serum Homocysteine in Males and Females with Coronary Heart Disease of Different Age Groups

Abstract: Problem statement: Elevated plasma total homocysteine concentration is a risk factor of cardiovascular disease. Total homocysteine level is a strong predictor of mortality in-patient with an angiographically confirmed Coronary Heart Disease (CHD), so we want to know at what age in males and females elevated homocysteine will be significa ntly elevated and this will help for better management and prognosis by decreasing the level of homocysteine. Approach: Plasma level of homocysteine was determined in male and female patients below and above 50 years old, who have coronary heart disease with diabetes or without dia betes. Fifty two coronary heart diseases with type 2 diabetic patients and a matched number of healthy s ubjects as a control and another 52 coronary heart disease patients without diabetes were included in this study. Plasma homocysteine was determined by Enzymatic Immuno Sorbant Assay (ELISA). Results: Plasma homocysteine level in coronary heart disease diabetic male and female patients who are 50 years old, respectively. Plasma homocysteine level in coronary heart disease nondiabetic male and female patients who are 50 years old, respecti vely. Conclusion: It is concluded that plasma level of homocysteine is significantly elevated in diabetic coronary heart disease female patients above 50 yea rs old and significantly elevated in nondiabetic coronary heart disease males and female patients, t hus nondiabetic coronary heart disease male and female patients and diabetic coronary female patien ts are at high risk of vascular diseases. It is recommended that these patients may take supplementation of folate and vitamin B12 to reduce the level of homocysteine.

Expression, Purification and Activity Assay of the Recombinant Protein of Catechol-O-Methyltransferase from Chinese White Shrimp (Fenneropenaeus chinensis)

Abstract: Problem statement: We have previously cloned a gene of Chinese white shrimp Catechol O-Methyltransferase (designated Fc-COMT) and characterized the gene expression pattern. In this study, expression and purification as well as activity assay of the recombinant Fc-COMT was further conducted. Approach: Using pET-30a (+) as a prokaryotic expression vector, the recombinant Fc- COMT was expressed in the supernatant of Escherichia coli lysate and easily purified by His-Bind resin chromatography. SDS-PAGE analysis showed that the molecular mass of recombinant Fc-COMT was approximately 30,000 Da, in good agreement with the software-predicted molecular weight. The enzymatic activity of recombinant Fc-COMT was tested using Dihydroxybenzoic Acid (DHBAc) as a substrate. Results: The methyl products of DHBAc, Vanillic Acid (VA) and Isovanillic Acid (IVA), were detected in the enzymatic reaction mixture with recombinant Fc-COMT by High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS). Conclusion: The recombinant Fc-COMT has catalytic activity of transferring methyl group from S-Adenosyl-L-Methionine (SAM) to the 3' hydroxyl or 4' hydroxyl group of benzyl ring of DHBAc.

Potentials and challenges of Participation in Agricultural Biotechnology R and D: A Case for National Agricultural Research Capacity Building

Abstract: Problem statement: Scientists in National Agricultural Research Institutes and University faculties of agriculture and veterinary medicine were surveyed to explore their participation in agricultural biotechnology research in Nigeria. Multistage sampling was used. Approach: Two Federal universities and two state universities were randomly selected from a list of Federal and State universities. In addition to these, one university each was selected from the four universities of Technology and three Federal universities of Agriculture. Results: Forty three scientists were purposively selected based on participation in agricultural biotechnology research. Nine research institutes were purposively selected based on their mandates. A total of 105 scientists were selected from the research institutes, based on their participation in agricultural biotechnology research. The total number of respondents from the selected amounted to 148. A structured questionnaire was used. Majority of the respondents fell within the medium participation category (63.5%). There is a significant relationship between availability of training/self development opportunities (r = 0.278, p<0.05) and career advancement opportunities (r = 0.348, p<0.05) and participation in agricultural biotechnology research. Inadequacy of electricity supply was significantly related to participation. Regression analysis show that human resources capacities available to scientists had positive relationship with participation in agricultural biotechnology research and development but only career advancement opportunities (r = 0.003, p<0.05) and royalties on findings (r = 0.151, p<0.05) were significant. Conclusion/Recommendations: There is need for strengthening both human resources and infrastructural capacity to increase participation in agricultural biotechnology research in Nigeria.

Tailor-Made Enzyme Carriers: Preparation and Use of Adsorbents Specifically Designed to Immobilize Allosteric Enzymes in Activated Conformation

Abstract: Problem statement: The enzyme immobilization has experienced substantial growth in the recent past and an ever increasing amount of study has been reported on various aspects of immobilized enzymes. In most of these investigations, catalytic activities are found to be diminished as compared to the enzyme free in solution. Approach: Hydrophobic adsorbents were prepared containing L-leucine or citric acid, two positive allosteric effectors, for bovine liver Glutamate Dehydrogenase (GDH, EC 1.4.1.3) and heart mitochondrial Malate Dehydrogenase (MDH, EC 1.1.1.37 ), respectively. Results: Immobilized preparations of these well-defined allosteric enzymes indicated improved catalytic activities as compared with those involving use of the adsorbents without these activators. Conclusion/Recommendations: It is concluded that the regulatory proteins are Furthermore; they retain their natural capacity for undergoing the conformational transitions needed for enhanced catalytic activities. Adsorptive immobilization of these two allosteric proteins in activated conformation may serve as useful models in relation to design strategies for preparation of tailor-made enzyme carriers.

Uterus-Relaxing Study of a Sudanese Herb (El-Hazha)

Abstract: Problem statement: The aim of this study is to investigate the pharmacological effects of the Methanolic-extract (AH2) of El-Hazha and its sub-fractions. Approach: These investigations were carried out on in vitro isolated uterus preparations from Non-Pregnant (NP) and Late-Pregnant rats (LP). In parallel displacement radio-ligand binding assay was performed for β-Adrenergic Receptors (β-ADR). Results: Showed that the herb and its different fractions produced dose-dependent relaxant effect (p<0.05, t-test, n = 6) on uterine contraction elicit by 25 mM KCl in both NP and LP uteri that affected significantly by fractionation, however, the effect of the most active fraction (AH2-11) was reversed by prior addition of propranolol (non-specific β-antagonist), but not affected by progesterone pre treatment of the LP rats. In addition, AH2 only in high concentration displaced isotopes from β-ADR, this affinity changed markedly by fractionation. Conclusion: We validate the fractionation effect on its relaxant activity and found partial role for β-ADR on mediating this activity. Future study was recommended to isolate and investigate its active components to enhance this activity or to discover a new novel natural therapeutic agent(s).