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.
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.
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.
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.
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.
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.
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.
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 (p0.05) 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.
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.
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.
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.