Commenced in January 2007
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Edition: International
Paper Count: 25

Search results for: glutamate dehydrogenase

25 New Malate Dehydrogenase-Glutamate Oxaolacetate Aminotransferase Glutamate Oxaloacetate Aminotransferase Enzyme System from Cereals and its Bioengineering Application

Authors: Zhanar S. Kudiyarova, Zhanar K. Rakhmetova, L. K. Bekbayeva, N. Z. Omirbekova, M. K. Gilmanov

Abstract:

Malate dehydrogenase-glutamate oxaloacetate aminotransferase (MDh-GOAT) enzyme complex (the EC) was isolated and purified from wheat and rise, their some main physicchemical properties were studied. Michael-s constants of the EC MDh-GOAT to malate, glutamate and NAD were investigated. This kinetic results show a high relationship to glutamate. Taking into account important role of the the EC in catabolism of glutamate – the central amino acid of a nitric exchange, there is a sharp necessity of deeper studying of this enzyme complex. Therefore the basic purpose of the work is studying the basic physical and chemical properties of this enzyme complex discovered by us, which would be very important for understanding the mechanisms of reaction catalyzed by the EC.

Keywords: Malate dehydrogenase-glutamate oxaloacetate aminotransferase, enzyme complex, glutamate.

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24 Glutamate Dehydrogenase and the Changing Pattern of Excretory Ammonia and Urea in Heteropneustes fossilis

Authors: Shuvasish Roy Choudhury, Rita Mahanta, Aparajita Borkotoki

Abstract:

Fishes, in general, follow ammonotelic mode of excretion. However, certain stress factors may provoke them to excrete urea. In the present study, the possible role of ureogenesis to avoid accumulation of toxic ammonia under water-restricted condition was tested in Heteropneustes fossilis. A total of hundred fishes were collected and sacrificed. Excretory urea and ammonia were estimated in the water of the aquarium and glutamate dehydrogenase acitivity was measured in the hepatic tissue. During the experimental period, excretory ammonia in Heteropneustes fossilis was found between 931% to 16% above the baseline ammonia and excretory urea was found between 112% to 898% above the baseline urea. A high degree of correlation with r (coefficient of correlation) above 0.9 is observed between excretory ammonia and urea in Heteropneustes fossilis. However, only a moderate degree of correlation is observed between the activity of glutamate dehydrogenase and excretory ammonia and urea.

Keywords: Ammonia, aquarium, glutamate dehydrogenase, urea, ureogenesis.

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23 Estimating Enzyme Kinetic Parameters from Apparent KMs and Vmaxs

Authors: Simon Brown, Noorzaid Muhamad, David C Simcock

Abstract:

The kinetic properties of enzymes are often reported using the apparent KM and Vmax appropriate to the standard Michaelis-Menten enzyme. However, this model is inappropriate to enzymes that have more than one substrate or where the rate expression does not apply for other reasons. Consequently, it is desirable to have a means of estimating the appropriate kinetic parameters from the apparent values of KM and Vmax reported for each substrate. We provide a means of estimating the range within which the parameters should lie and apply the method to data for glutamate dehydrogenase from the nematode parasite of sheep Teladorsagia circumcincta.

Keywords: enzyme kinetics, glutamate dehydrogenase, intervalanalysis, parameter estimation.

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22 Development of a Brain Glutamate Microbiosensor

Authors: Kartika S. Hamdan, Zainiharyati M. Zain, Mohamed I. A. Halim, Jafri M. Abdullah, Robert D. O'Neill

Abstract:

This work attempts to improve the permselectivity of poly-ortho-phenylenediamine (PPD) coating for glutamate biosensor applications on Pt microelectrode, using constant potential amperometry and cyclic voltammetry. Percentage permeability of the modified PPD microelectrode was carried out towards hydrogen peroxide (H2O2) and ascorbic acid (AA) whereas permselectivity represents the percentage interference by AA in H2O2 detection. The 50-μm diameter Pt disk microelectrode showed a good permeability value toward H2O2 (95%) and selectivity against AA (0.01%) compared to other sizes of electrode studied here. The electrode was further modified with glutamate oxidase (GluOx) that was immobilized and cross linked with glutaraldehyde (GA, 0.125%), resulting in Pt/PPD/GluOx-GA electrode design. The maximum current density Jmax and apparent Michaelis constant, KM, obtained on Pt/PPD/GluOx-GA electrodes were 48 μA cm-2 and 50 μM, respectively. The linear region slope (LRS) was 0.96 μA cm-2 mM-1. The detection limit (LOD) for glutamate was 3.0 ± 0.6 μM. This study shows a promising glutamate microbiosensor for brain glutamate detection. 

Keywords: Brain, Glutamate, Microbiosensor.

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21 Novel Structural Insights of Glutamate Racemase from Mycobacterium tuberculosis through Modeling and Docking Studies

Authors: Jayashree Ramana

Abstract:

An alarming emergence of multidrug-resistant strains of the tuberculosis pathogen Mycobacterium tuberculosis and continuing high worldwide incidence of tuberculosis has invigorated the search for novel drug targets. The enzyme glutamate racemase (MurI) in bacteria catalyzes the stereoconversion of L-glutamate to D-glutamate which is a component of the peptidoglycan cell wall of the bacterium. The inhibitors targeted against MurI from several bacterial species have been patented and are advocated as promising antibacterial agents. However there are none available against MurI from Mycobacterium tuberculosis, due to the lack of its threedimensional structure. This work accomplished two major objectives. First, the tertiary structure of MtMurI was deduced computationally through homology modeling using the templates from bacterial homologues. It is speculated that like in other Gram-positive bacteria, MtMurI exists as a dimer and many of the protein interactions at the dimer interface are also conserved. Second, potent candidate inhibitors against MtMurI were identified through docking against already known inhibitors in other organisms.

Keywords: Glutamate racemase, homology modeling, docking, drug resistance.

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20 Influence of Hydrocarbons on Plant Cell Ultrastructure and Main Metabolic Enzymes

Authors: T. Sadunishvili, E. Kvesitadze, M. Betsiashvili, N. Kuprava, G. Zaalishvili, G. Kvesitadze

Abstract:

Influence of octane and benzene on plant cell ultrastructure and enzymes of basic metabolism, such as nitrogen assimilation and energy generation have been studied. Different plants: perennial ryegrass (Lolium perenne) and alfalfa (Medicago sativa); crops- maize (Zea mays L.) and bean (Phaseolus vulgaris); shrubs – privet (Ligustrum sempervirens) and trifoliate orange (Poncirus trifoliate); trees - poplar (Populus deltoides) and white mulberry (Morus alba L.) were exposed to hydrocarbons of different concentrations (1, 10 and 100 mM). Destructive changes in bean and maize leaves cells ultrastructure under the influence of benzene vapour were revealed at the level of photosynthetic and energy generation subcellular organells. Different deviations at the level of subcellular organelles structure and distribution were observed in alfalfa and ryegrass root cells under the influence of benzene and octane, absorbed through roots. The level of destructive changes is concentration dependent. Benzene at low 1 and 10 mM concentration caused the increase in glutamate dehydrogenase (GDH) activity in maize roots and leaves and in poplar and mulberry shoots, though to higher extent in case of lower, 1mM concentration. The induction was more intensive in plant roots. The highest tested 100mM concentration of benzene was inhibitory to the enzyme in all plants. Octane caused induction of GDH in all grassy plants at all tested concentrations; however the rate of induction decreased parallel to increase of the hydrocarbon concentration. Octane at concentration 1 mM caused induction of GDH in privet, trifoliate and white mulberry shoots. The highest, 100mM octane was characterized by inhibitory effect to GDH activity in all plants. Octane had inductive effect on malate dehydrogenase in almost all plants and tested concentrations, indicating the intensification of Trycarboxylic Acid Cycle. The data could be suggested for elaboration of criteria for plant selection for phytoremediation of oil hydrocarbons contaminated soils.

Keywords: Higher plants, hydrocarbons, cell ultrastructure, glutamate and malate dehydrogenases.

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19 Biochemical Changes in the Liver of Mice after Exposure to Different Doses of Diclofenac Sodium

Authors: Deepak Mohan, Sushma Sharma

Abstract:

Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) are a group of widely used drugs for the treatment of rheumatoid diseases and to relieve pain and inflammation due to their analgesic anti-pyretic and anti-inflammatory properties. The therapeutic and many of the toxic effects of NSAIDs result from reversible inhibition of enzymes in the cyclooxygenase (COX) group. In the present investigation the effect of the drug on the concentration of lipids, and on the activity of the enzymes i.e. acid and alkaline phosphatase, GOT, GPT and lipid peroxidase were studied. There was a significant enhancement in the activities of both acid and alkaline phosphatase after 21 days of treatment. Proportionate increase in the MDA contents was observed after different days of diclofenac treatment. Cellular damage in the liver resulted in decrease in the activity of both GOT (Glutamate oxaloacetate transaminase) and GPT (Glutamate pyruvate transaminase) in both low and high dose groups. Significant decrease in the liver contents was also observed in both dose groups.

Keywords: Anti-inflammatory, cyclooxygenase, glutamate oxaloacetate transaminase, malondialdehyde.

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18 Impact of Herbicides on Soil Biology in Rapeseed

Authors: M. Eickermann, M. K. Class, J. Junk

Abstract:

Winter oilseed rape, Brassica napus L., is characterized by a high number of herbicide applications. Therefore, its cultivation can lead to massive contamination of ground water and soil by herbicide and their metabolites. A multi-side long-term field experiment (EFFO, Efficient crop rotation) was set-up in Luxembourg to quantify these effects. Based on soil sampling and laboratory analysis, preliminary results showed reduced dehydrogenase activities of several soil organisms due to herbicide treatments. This effect is highly depending on the soil type. Relation between the dehydrogenase activity and the amount of microbial carbon showed higher variability on the test side with loamy Brown Earth, based on Bunter than on those with sandy-loamy Brown Earth, based on calciferous Sandstone.

Keywords: Cropping system, dehydrogenase activity, herbicides, mechanical weed control, oilseed rape.

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17 Protective Effect of Ethanolic Extract of Polyherbal Formulation on Carbon Tetrachloride Induced Liver Injury

Authors: R. Kokilavani, K. Gurusamy, K. Arumugasamy

Abstract:

Protective effect of ethanolic extract of polyherbal formulation (PHF) was studied on carbon tetrachloride induced liver damage on carbon tetrachloride induced liver damage. Treatment of rats with 250mg /kg body weight of ethanolic extract of PHF protected rats against carbon tetrachloride liver injury by significant lowerering 5’ nucleotidase (5’NT), Gamma Glutamyl transferase (GGT), Glutamate dehdyrogenasse (GDH) and Succinate Dehydrogenase (SDH) levels compared to control. Normalization in these enzyme levels indicates strong hepatoprotective property of PHF extract.

Keywords: Carbon tetrachloride, ethanolic, hepatoprotective, polyherbal formulation.

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16 Gamma Glutamyl Transferase and Lactate Dehydrogenase as Biochemical Markers of Severity of Preeclampsia

Authors: S. M. Munde, N. R. Hazari, A. P. Thorat, S. B. Gaikwad, V. S. Hatolkar

Abstract:

This study was conducted to examine the possible role of serum Gamma-glutamyltransferase (GGT) and Lactate dehydrogenase (LDH) in the prediction of severity of preeclampsia. The study group comprised of 40 preeclamptic cases (22 with mild and 18 with severe) and 40 healthy normotensive pregnant controls. Serum samples of all the cases were assayed for GGT and LDH. Demographic, hemodynamic and laboratory data as well as serum GGT and LDH levels were compared among the three groups.

The results indicated that severe preeclamptic cases had significantly increased levels of serum GGT and LDH. The symptoms in severe preeclamptic women were significantly increased in patients with GGT > 70 IU/L and LDH >800 IU/L. Elevated levels of serum GGT and LDH can be used as biochemical markers which reflects the severity of preeclampsia and useful for the management of preeclampsia to decrease maternal and fetal morbidity and mortality.

Keywords: Severe Preeclampsia, GGT, LDH.

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15 Gonadotoxic and Cytotoxic Effect of Induced obesity via Monosodium Glutamate on Mus musculus Testis Cytoarchitecture and Sperm Parameter

Authors: I. Nur Hilwani, R. Nasibah, S. Nurdiana, M. J. Norashirene

Abstract:

Impaired fertility may be the result of indirect consumption of anti-fertility agents through food. Monosodium glutamate (MSG) has been widely used as food additive, flavour enhancer and included in vaccines. This study focuses in determining the gonadotoxic and cytotoxic effect of MSG on selected sperm parameters such as sperm viability, sperm membrane integrity and testes cytoarchitecture of male mice via histological examination to determine its effect on spermatogenesis. Twenty-four Mus musculus were randomly divided into 4 groups and given intraperitoneal injections (IP) daily for 14 days of different MSG concentrations at 250, 500 and 1000mg/kg MSG to body weight to induce obesity. Saline was given to control group. Mice were sacrificed and analysis revealed abnormalities in values for sperm parameters and damages to testes cytoarchitecture of male mice. The results recorded decreased viability (p<0.05) and integrity of sperm membrane (p>0.05) with degenerative structures in seminiferous tubule of testes. The results indicated various implications of MSG on male mice reproductive system which has consequences in fertility potential.

Keywords: Sperm parameter, sperm viability, sperm membrane integrity and testes histology.

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14 Effects of Allelochemical Gramine on Metabolic Activity and Ultrastructure of Cyanobacterium Microcystis aeruginosa

Authors: Y. Hong, H. Y. Hu, A. Sakoda, M. Sagehashi

Abstract:

In this study, inhibition of Microcystis aeruginosa by antialgal alleochemical gramine, was studied by analyzing algal metabolic activity (represented by esterase and total dehydrogenase activities) and cell ultrastructure (showing morphological and ultrastructure alterations using transmission electron microscopy and DNA ladder analysis). After gramine exposure, esterase and total dehydrogenase activities were increased firstly but decreased later. In contrast with the controls, the cells exposed to gramine showed apparent ultrastructure alterations with thylakoids in breakage, phycobilins in decrease, lipid and cyanophycin granules abundant firstly but dissolved afterwards, DNA in fragementation. The occurrence of increase of metabolic activity and specific granules reflected that the resistance of cellular response to gramine was initiated. DNA fragementation associated with the increase of metabolic activity and specific granules hinted that gramine caused M. aeruginosa cells to initiate some morphotype of programmed cell death.

Keywords: Allelochemical, gramine, metabolic activity, Microcystis aeruginosa, ultrastructure.

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13 Effects of Intrauterine and Extrauterine Exposure to 1800 MHz GSM-Like Radiofrequency Radiation on Liver Regulatory Enzymes Activities in Infant Female Rabbits

Authors: A. Tomruk, G. Guler, B. Tandogan, E. Ozgur, N.N. Ulusu, N. Seyhan

Abstract:

In the present study, we aimed to design the intrauterine and extrauterine exposure to 1800 MHz GSM-like RF radiation and investigate its possible bio-effects on infant female rabbits. Totally thirty-six New Zealand White female rabbits, onemonth old, were randomly divided into four groups which are composed of 9 rabbits; i. Group I [Intrauterine (IU) exposure(-); Extrauterine (EU) exposure (-)], Group II [IU exposure (-); EU exposure (+)], Group III [IU exposure(+);EU exposure(-)], Group IV [IU exposure (+);EU exposure(+)]. The master regulatory enzymes activities of pentose phosphate pathway (glucose-6-phosphate dehydrogenase, G-6PD; 6-phosphogluconate dehydrogenase, 6- PGDH) and glutathione-dependent metabolism (glutathione peroxidase, GSH-Px; glutathione reductase, GR; glutathione Stransferase, GST, thioredoxin reductase, TRx) were analyzed in liver tissues of young female rabbits. Decreased G-6PD, 6-PGD, GSH-Px, GR activities were found in Group III compared to Group I (p<0.05, Mann Whitney). Increased GSH-px and TRx activities were found in Group IV compared to Group I (p<0.05, Mann Whitney). It can be concluded that the intrauterine and extrauterine exposure to GSMlike RF radiation may influence the liver regulatory enzymes activities.

Keywords: Radiofrequency (RF), intrauterine (IU) andextrauterine (EU) exposure, infant female rabbits.

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12 Stabilizing Effects of Deep Eutectic Solvents on Alcohol Dehydrogenase Mediated Systems

Authors: Fatima Zohra Ibn Majdoub Hassani, Ivan Lavandera, Joseph Kreit

Abstract:

This study explored the effects of different organic solvents, temperature, and the amount of glycerol on the alcohol dehydrogenase (ADH)-catalysed stereoselective reduction of different ketones. These conversions were then analyzed by gas chromatography. It was found that when the amount of deep eutectic solvents (DES) increases, it can improve the stereoselectivity of the enzyme although reducing its ability to convert the substrate into the corresponding alcohol. Moreover, glycerol was found to have a strong stabilizing effect on the ADH from Ralstonia sp. (E. coli/ RasADH). In the case of organic solvents, it was observed that the best conversions into the alcohols were achieved with DMSO and hexane. It was also observed that temperature decreased the ability of the enzyme to convert the substrates into the products and also affected the selectivity. In addition to that, the recycling of DES up to three times gave good conversions and enantiomeric excess results and glycerol showed a positive effect in the stability of various ADHs. Using RasADH, a good conversion and enantiomeric excess into the S-alcohol were obtained. It was found that an enhancement of the temperature disabled the stabilizing effect of glycerol and decreased the stereoselectivity of the enzyme. However, for other ADHs a temperature increase had an opposite positive effect, especially with ADH-T from Thermoanaerobium sp. One of the objectives of this study was to see the effect of cofactors such as NAD(P) on the biocatlysis activities of ADHs.

Keywords: Alcohol dehydrogenases, DES, gas chromatography, RasADH.

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11 Determination of Effective Variables on Arachidonic Acid Production by Mortierella alpina CBS 754.68in Solid-State Fermentation using Plackett-Burman Screening Design

Authors: Z. Ghobadi, Z. Hamidi- Esfahani, M. H. Azizi

Abstract:

In the present study, the oleaginous fungus Mortierella alpina CBS 754.68 was screened for arachidonic acidproduction using inexpensive agricultural by-products as substrate. Four oilcakes were analysed to choose the best substrate among them. Sunflower oilcake was the most effective substrate for ARA production followed by soybean, colza and olive oilcakes. In the next step, seven variables including substrate particle size, moisture content, time, temperature, yeast extract supply, glucose supply and glutamate supply were surveyed and effective variables for ARA production were determined using a Plackett-Burman screening design. Analysis results showed that time (12 days), substrate particle size (1-1.4 mm) and temperature (20ºC) were the most effective variables for the highest level of ARA production respectively.

Keywords: Arachidonic acid, Mortierella alpine, Solid-statefermentation, Plackett-Burman design

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10 Effect of Different Fertilization Methods on Soil Biological Indexes

Authors: Khosro Mohammadi

Abstract:

Fertilization plays an important role in crop growth and soil improvement. This study was conducted to determine the best fertilization system for wheat production. Experiments were arranged in a complete block design with three replications in two years. Main plots consisted of six methods of fertilization including (N1): farmyard manure; (N2): compost; (N3): chemical fertilizers; (N4): farmyard manure + compost; (N5): farmyard manure + compost + chemical fertilizers and (N6): control were arranged in sub plots. The addition of compost or farm yard manure significantly increased the soil microbial biomass carbon in comparison to the chemical fertilizer. The dehydrogenase, phosphatase and urease activities in the N3 treatment were significantly lower than in the farm yard manure and compost treatments.

Keywords: Enzyme activity, fertilization, microbial biomasscarbon, wheat.

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9 Pollution Induced Community Tolerance(PICT) of Microorganisms in Soil Incubated with Different Levels of PB

Authors: N. Aliasgharzad, A. Molaei, S. Oustan

Abstract:

Soil microbial activity is adversely affected by pollutants such as heavy metals, antibiotics and pesticides. Organic amendments including sewage sludge, municipal compost and vermicompost are recently used to improve soil structure and fertility. But, these materials contain heavy metals including Pb, Cd, Zn, Ni and Cu that are toxic to soil microorganisms and may lead to occurrence of more tolerant microbes. Among these, Pb is the most abundant and has more negative effect on soil microbial ecology. In this study, Pb levels of 0, 100, 200, 300, 400 and 500 mg Pb [as Pb(NO3)2] per kg soil were added to the pots containing 2 kg of a loamy soil and incubated for 6 months at 25°C with soil moisture of - 0.3 MPa. Dehydrogenase activity of soil as a measure of microbial activity was determined on 15, 30, 90 and 180 days after incubation. Triphenyl tetrazolium chloride (TTC) was used as an electron acceptor in this assay. PICTs (€IC50 values) were calculated for each Pb level and incubation time. Soil microbial activity was decreased by increasing Pb level during 30 days of incubation but the induced tolerance appeared on day 90 and thereafter. During 90 to 180 days of incubation, the PICT was gradually developed by increasing Pb level up to 200 mg kg-1, but the rate of enhancement was steeper at higher concentrations.

Keywords: Induced tolerance, soil microorganisms, Pb, PICT, pollutants.

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8 Charaterisation of Salmonella Isolated from Nile Tilapia (Oreochromis niloticus) along Lake Victoria Beaches in Western Kenya

Authors: Wandili S. Awuor, Onyango D. Miruka, Waindi N. Eliud

Abstract:

Foodborne Salmonella infections have become a major problem world wide. Salmonellosis transmitted from fish are quite common. Established quality control measures exist for export oriented fish, none exists for fish consumed locally. This study aimed at characterization of Salmonella isolated from Nile tilapia . The study was carried out in selected beaches along L. Victoria in Western Kenya between March and June 2007. One hundred and twenty fish specimens were collected. Salmonella isolates were confirmed using serotyping, biochemical testing in addition to malic acid dehydrogenase (mdh) and fliC gene sequencing. Twenty Salmonella isolates were confirmed by mdh gene sequencing. Nine (9) were S. enterica serotype typhimurium, four (4) were S. enterica Serotype, enteritidis and seven (7) were S. enterica serotype typhi. Nile tilapia have a role in transmission of Salmonellosis in the study area, poor sanitation was a major cause of pollution at the beach inshore waters.

Keywords: fliC, mdh, Salmonellosis, Serotype

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7 Enzymes Activity in Bovine Cervical Mucus Related to the Time of Ovulation And Insemination

Authors: S. Benbia, A.Kalla, M. Yahia, K. Belhadi, A. Zidani

Abstract:

Forty-five dairy cows were used to compare the enzyme activity of alkaline phosphatase (ALP), lactate dehydrogenase (LDH), α -amylase in the cervical mucus of cows during spontaneous and induced estrus using progestagen or PGF2 α and to determine whether these enzymes affect the fertility in cows with induced estrus, at the time of Al. The animals were assigned to 3 groups (no treatment, a Crestar® for 12 days, a double im injection of PGF2 α). The cows were artificially inseminated (AI). Cervical mucus samples were collected from all cows 3 to 5 min before the AI. The results are summarized as follows: ALP and α -amylase activity for spontaneous estrus were similar to those for induced estrus (P>0.05) . LDH activity levels during spontaneous and PGF2 α induced estrus was significantly lower (P < 0.001) than that in progestagene induced estrus groups. While no difference was found between the first and the third groups. Our result showed a significant difference in LDH activity levels between cows conceived with 2 or more AI and those conceived with 1 AI. The result of this study showed that the enzyme activity in cervical mucus is helpful for detection of ovulation and time of AI.

Keywords: cervical mucus, dairy cow, enzyme, induced, estrus, ovulation, AI

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6 The Role of Glutamine-Rich Region of Candida Albicans Tec1p in Mediating Morphological Transition and Invasive Growth

Authors: W. Abu Rayyan, A. Singh, A. M. Al-Jaafreh, W. Abu Dayyih, M. Bustami, S. Salem, N. Seder, K. Schröppel

Abstract:

Hyphal growth and the transcriptional regulation to the host environment are key issues during the pathogenesis of C. albicans. Tec1p is the C. albicans homolog of a TEA transcription factor family, which share a conserved DNA-binding TEA domain in their N-terminal. In order to define a structure-function relationship of the C. albicans Tec1p protein, we constructed several mutations on the N terminal, C terminal or in the TEA binding domain itself by homologous recombination technology. The modifications in the open reading frame of TEC1 were tested for reconstitution of the morphogenetic development of the tec1/tec1 mutant strain CaAS12. Mutation in the TEA consensus sequence did not confer transition to hyphae whereas the reconstitution of the full-length Tec1p has reconstituted hyphal development. A deletion in one of glutamine-rich regions either in the Tec1p N-terminal or the C-terminal in regions of 53-212 or 637–744 aa, respectively, did not restore morphological development in mutant CaAS12 strain. Whereas, the reconstitution with Tec1p mutants other than the glutamate-rich region has restored the morphogenetic switch. Additionally, the deletion of the glutamine-rich region has attenuated the invasive growth and the heat shock resistance of C. albicans. In conclusion, we show that a glutamine-rich region of Tec1p is essential for the hyphal development and mediating adaptation to the host environment of C. albicans.

Keywords: Candida albicans, transcription factor, TEA domain, hyphal formation, morphogenetic development, TEC1, Tet-induced.

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5 Apoptosis Activity of Persea declinata (Bl.) Kosterm Bark Methanolic Crude Extract

Authors: P. Narrima, C. Y. Looi, M. A. Mohd, H. M. Ali

Abstract:

Persea declinata (Bl.) Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill), which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl.) Kosterm bark methanolic crude extract (PDM). PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 .g/mL after 48h of treatment. We observed that PDM caused cell cycle arrest and subsequent apoptosis in MCF-7 cells, as exhibited by increased population at G0/G1 phase, higher lactate dehydrogenase (LDH) release, and DNA fragmentation. Mechanistic studies showed that PDM caused significant elevation in ROS production, leading to perturbation of mitochondrial membrane potential, cell permeability, and activation of caspases-3/7. On the other hand, real-time PCR and Western blot analysis showed that PDM treatment increased the expression of the proapoptotic molecule, Bax, but decreased the expression of prosurvival proteins, Bcl-2 and Bcl-xL, in a dose-dependent manner. These findings imply that PDM could inhibit proliferation in MCF-7 cells via cell cycle arrest and apoptosis induction, indicating its potential as a therapeutic agent worthy of further development.

Keywords: Antiproliferative, apoptosis, MCF-7 human breast cancer, Persea declinata.

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4 Higher Plants Ability to Assimilate Explosives

Authors: G. Khatisashvili, M. Gordeziani, G. Adamia, E. Kvesitadze, T. Sadunishvili, G. Kvesitadze

Abstract:

The ability of agricultural and decorative plants to absorb and detoxify TNT and RDX has been studied. All tested 8 plants, grown hydroponically, were able to absorb these explosives from water solutions: Alfalfa > Soybean > Chickpea> Chikling vetch >Ryegrass > Mung bean> China bean > Maize. Differently from TNT, RDX did not exhibit negative influence on seed germination and plant growth. Moreover, some plants, exposed to RDX containing solution were increased in their biomass by 20%. Study of the fate of absorbed [1-14ðí]-TNT revealed the label distribution in low and high-molecular mass compounds, both in roots and above ground parts of plants, prevailing in the later. Content of 14ðí in lowmolecular compounds in plant roots are much higher than in above ground parts. On the contrary, high-molecular compounds are more intensively labeled in aboveground parts of soybean. Most part (up to 70%) of metabolites of TNT, formed either by enzymatic reduction or oxidation, is found in high molecular insoluble conjugates. Activation of enzymes, responsible for reduction, oxidation and conjugation of TNT, such as nitroreductase, peroxidase, phenoloxidase and glutathione S-transferase has been demonstrated. Among these enzymes, only nitroreductase was shown to be induced in alfalfa, exposed to RDX. The increase in malate dehydrogenase activities in plants, exposed to both explosives, indicates intensification of Tricarboxylic Acid Cycle, that generates reduced equivalents of NAD(P)H, necessary for functioning of the nitroreductase. The hypothetic scheme of TNT metabolism in plants is proposed.

Keywords: Higher plants, TNT, RDX, transformation.

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3 Differential Sensitivity of Nitrogen-Fixing, Filamentous Cyanobacterial Species to an Organochlorine Insecticide - 6, 7, 8, 9, 10, 10- Hexachloro-1, 5, 5a, 6, 9, 9a-Hexahydro-6, 9- Methano-2, 4, 3-Benzodioxathiepine-3-Oxide

Authors: Nirmal J.I. Kumar, Anubhuti A. Bora, Manmeet K. Amb

Abstract:

Application of pesticides in the paddy fields has deleterious effects on non-target organisms including cyanobacteria which are photosynthesizing and nitrogen fixing micro-organisms contributing significantly towards soil fertility and crop yield. Pesticide contamination in the paddy fields has manifested into a serious global environmental concern. To study the effect of one such pesticide, three cyanobacterial strains; Anabaena fertilissima, Aulosira fertilissima and Westiellopsis prolifica were selected for their stress responses to an Organochlorine insecticide - 6, 7, 8, 9, 10, 10-hexachloro-1, 5, 5a, 6, 9, 9a-hexahydro-6, 9-methano-2, 4, 3- benzodioxathiepine-3-oxide, with reference to their photosynthesic pigments-chlorophyll-a and carotenoids as well as accessory pigments-phycobiliproteins (phycocyanin, allophycocyanin and phycoerythrin), stress induced biochemical metabolites like carbohydrates, proteins, amino acids, phenols and enzymes-nitrate reductase, glutamine synthetase and succinate dehydrogenase. All the three cyanobacterial strains were adversely affected by the insecticide doses and inhibition was dose dependent. Reduction in photosynthetic and accessory pigments, metabolites, nitrogen fixing and respiratory enzymes of the test organisms were accompanied with an initial increase in their total protein at lower Organochlorine doses. On the other hand, increased amount of phenols in all the insecticide treated concentrations was indicative of stressed activities of the organisms.

Keywords: biochemical metabolites, endosulfan, enzymes, pigments

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2 ALDH1A1 as a Cancer Stem Cell Marker: Value of Immunohistochemical Expression in Benign Prostatic Hyperplasia, Prostatic Intraepithelial Neoplasia, and Prostatic Adenocarcinoma

Authors: H. M. Abdelmoneim, N. A. Babtain, A. S. Barhamain, A. Z. Kufiah, A. S. Malibari, S. F. Munassar, R. S. Rawa

Abstract:

Introduction: Prostate cancer is one of the most common causes of morbidity and mortality in men in developed countries. Cancer Stem Cells (CSCs) could be responsible for the progression and relapse of cancer. Therefore, CSCs markers could provide a prognostic strategy for human malignancies. Aldehyde dehydrogenase 1A1 (ALDH1A1) activity has been shown to be associated with tumorigenesis and proposed to represent a functional marker for tumor initiating cells in various tumor types including prostate cancer. Material & Methods: We analyzed the immunohistochemical expression of ALDH1A1 in benign prostatic hyperplasia (BPH), prostatic intraepithelial neoplasia (PIN) and prostatic adenocarcinoma and assessed their significant correlations in 50 TURP sections. They were microscopically interpreted and the results were correlated with histopathological types and tumor grade. Results: In different prostatic histopathological lesions we found that ALDH1A1 expression was low in BPH (13.3%) and PIN (6.7%) and then its expression increased with prostatic adenocarcinoma (40%), and this was statistically highly significant (P value = 0.02). However, in different grades of prostatic adenocarcinoma we found that the higher the Gleason grade the higher the expression for ALDH1A1 and this was statistically significant (P value = 0.02). We compared the expression of ALDH1A1 in PIN and prostatic adenocarcinoma. ALDH1A1 expression was decreased in PIN and highly expressed in prostatic adenocarcinoma and this was statistically significant (P value = 0.04). Conclusion: Increasing ALDH1A1 expression is correlated with aggressive behavior of the tumor. Immunohistochemical expression of ALDH1A1 might provide a potential approach to study tumorigenesis and progression of primary prostate carcinoma.

Keywords: ALDH1A1, BPH, PIN, prostatic adenocarcinoma.

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1 Enzyme Involvement in the Biosynthesis of Selenium Nanoparticles by Geobacillus wiegelii Strain GWE1 Isolated from a Drying Oven

Authors: Daniela N. Correa-Llantén, Sebastián A. Muñoz-Ibacache, Mathilde Maire, Jenny M. Blamey

Abstract:

The biosynthesis of nanoparticles by microorganisms, on the contrary to chemical synthesis, is an environmentally-friendly process which has low energy requirements. In this investigation, we used the microorganism Geobacillus wiegelii, strain GWE1, an aerobic thermophile belonging to genus Geobacillus, isolated from a drying oven. This microorganism has the ability to reduce selenite evidenced by the change of color from colorless to red in the culture. Elemental analysis and composition of the particles were verified using transmission electron microscopy and energy-dispersive X-ray analysis. The nanoparticles have a defined spherical shape and a selenium elemental state. Previous experiments showed that the presence of the whole microorganism for the reduction of selenite was not necessary. The results strongly suggested that an intracellular NADPH/NADH-dependent reductase mediates selenium nanoparticles synthesis under aerobic conditions. The enzyme was purified and identified by mass spectroscopy MALDI-TOF TOF technique. The enzyme is a 1-pyrroline-5-carboxylate dehydrogenase. Histograms of nanoparticles sizes were obtained. Size distribution ranged from 40-160 nm, where 70% of nanoparticles have less than 100 nm in size. Spectroscopic analysis showed that the nanoparticles are composed of elemental selenium. To analyse the effect of pH in size and morphology of nanoparticles, the synthesis of them was carried out at different pHs (4.0, 5.0, 6.0, 7.0, 8.0). For thermostability studies samples were incubated at different temperatures (60, 80 and 100 ºC) for 1 h and 3 h. The size of all nanoparticles was less than 100 nm at pH 4.0; over 50% of nanoparticles have less than 100 nm at pH 5.0; at pH 6.0 and 8.0 over 90% of nanoparticles have less than 100 nm in size. At neutral pH (7.0) nanoparticles reach a size around 120 nm and only 20% of them were less than 100 nm. When looking at temperature effect, nanoparticles did not show a significant difference in size when they were incubated between 0 and 3 h at 60 ºC. Meanwhile at 80 °C the nanoparticles suspension lost its homogeneity. A change in size was observed from 0 h of incubation at 80ºC, observing a size range between 40-160 nm, with 20% of them over 100 nm. Meanwhile after 3 h of incubation at size range changed to 60-180 nm with 50% of them over 100 nm. At 100 °C the nanoparticles aggregate forming nanorod structures. In conclusion, these results indicate that is possible to modulate size and shape of biologically synthesized nanoparticles by modulating pH and temperature.

Keywords: Genus Geobacillus, NADPH/NADH-dependent reductase, Selenium nanoparticles.

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