Probing the role of highly conserved residues in triosephosphate isomerase-analysis of site specific mutants at positions 64 and 75 in the Plasmodial enzyme

Highly conserved residues in enzymes are often found to be clustered close to active sites, suggesting that functional constraints dictate the nature of amino acid residues accommodated at these sites. Using the Plasmodiumfalciparum triosephosphate isomerase (PfTIM) enzyme () as a template, we have examined the effects of mutations at positions 64 and 75, which are not directly involved in the proton transfer cycle. Thr (T) occurring at position 75 is completely conserved, whereas only Gln (Q) and Glu (E) are accommodated at position 64. Biophysical and kinetic data are reported for four T75 (T75S/V/C/N) and two Q64 (Q64N/E) mutants. The dimeric structure is weakened in the Q64E and Q64N mutants, whereas dimer integrity is unimpaired in all four T75 mutants. Measurement of the concentration dependence of enzyme activity permits an estimate of K-d values for dimer dissociation (Q64N=73.79.2nm and Q64E=44.6 +/- 8.4nm). The T75S/V/C mutants have activities comparable to the wild-type enzyme, whereas a fourfold drop is observed for T75N. All four T75 mutants show a dramatic fall in activity between 35 degrees C and 45 degrees C. Crystal structure determination of the T75S/V/N mutants provides insights into the variations in local interactions, with the T75N mutant showing the largest changes. Hydrogen-bond interactions determine dimer stability restricting the choice of residues at position 64 to Gln (Q) and Glu (E). At position 75, the overwhelming preference for Thr (T) may be dictated by the imperative of maintaining temperature stability of enzyme activity.

Mapping Key Residues of ISD11 Critical for NFS1-ISD11 Subcomplex Stability IMPLICATIONS IN THE DEVELOPMENT OF MITOCHONDRIAL DISORDER, COXPD19

Biogenesis of the iron-sulfur (Fe-S) cluster is an indispensable process in living cells. In mammalian mitochondria, the initial step of the Fe-S cluster assembly process is assisted by the NFS1-ISD11 complex, which delivers sulfur to scaffold protein ISCU during Fe-S cluster synthesis. Although ISD11 is an essential protein, its cellular role in Fe-S cluster biogenesis is still not defined. Our study maps the important ISD11 amino acid residues belonging to putative helix 1 (Phe-40), helix 3 (Leu-63, Arg-68, Gln-69, Ile-72, Tyr-76), and C-terminal segment (Leu-81, Glu-84) are critical for in vivo Fe-S cluster biogenesis. Importantly, mutation of these conserved ISD11 residues into alanine leads to its compromised interaction with NFS1, resulting in reduced stability and enhanced aggregation of NFS1 in the mitochondria. Due to altered interaction with ISD11 mutants, the levels of NFS1 and Isu1 were significantly depleted, which affects Fe-S cluster biosynthesis, leading to reduced electron transport chain complex (ETC) activity and mitochondrial respiration. In humans, a clinically relevant ISD11 mutation (R68L) has been associated in the development of a mitochondrial genetic disorder, COXPD19. Our findings highlight that the ISD11 R68A/R68L mutation display reduced affinity to form a stable subcomplex with NFS1, and thereby fails to prevent NFS1 aggregation resulting in impairment of the Fe-S cluster biogenesis. The prime affected machinery is the ETC complex, which showed compromised redox properties, causing diminished mitochondrial respiration. Furthermore, the R68L ISD11 mutant displayed accumulation of mitochondrial iron and reactive oxygen species, leading to mitochondrial dysfunction, which correlates with the phenotype observed in COXPD19 patients.

The Characteristics of Semi-Cokes --The Solid Residues from Coal Partial Gasification

In this paper the proximate analysis and ultimate analysis of sulfur in different semi-cokes generated from Rizhao bituminous coal and Beijing anthracite under different temperatures is done. Also the tendency of the contents of volatile, ash, fixed carbon and sulfur in different semi-cokes along with the different preparation temperatures is studied. Then the combustion experiment of semi-cokes in the drop-tube furnace system was carried out, and the kinetic parameters of different semi-cokes ware calculated.

The analysis of dieldrin residues in the brain, liver and muscles of the African catfish Clarias gariepinus (Burchell 1822) chronically exposed to dieldrin

Mature adult Clarias gariepinus were obtained at the ABRU hatchery in Sonning (UK), where they had beenbred and reared for several years. These were exposed to two concentrations of dieldrin in water (2.4 mu g super(-1) and 4.0 mu g super(-1). The residue analysis of diedrin in three tissues exposed for on moth at two concentrations was carried out. These were subjected to GLC analytical process. The results indicated significantly (P<0.05) higher residues in liver than in muscle and brain. The results also showed that residue levels were dependant on exposure concentration

Limit of drug residues in aquatic products (NY5071-2002)

Weimin, ZOU, lexian YANG, lan JIANG, Shuqin WU, Qi YI, Jianli WU

Pesticide residues in marine fishes

Pesticide residues in four marine fishes, black pomfret (Parastromateus niger), mackerel (Rastrelliger kanagurta), marine vala (Chirocentrus sp.) and tuna (Euthynnus affinis) are reported. Highest concentration is found in black pomfret followed by tuna, vala and mackerel. The contents of various pesticides present in fish under study are well below the action level prepared by FDA to cause any health hazard.

Organochlorine insecticide residues in sediments from the coast of Chittagong, Bangladesh

A total of 5 samples of marine sediments were analyzed for residues of organochlorine insecticide from the coast of Chittagong, Bangladesh. The analytical method consisted of 3 phases, extraction, clean-up and, analysis through Gas Chromatography (GC) with Electron Capture Detector (ECD). The concentration ranges were as follows 0.18 - 1.33 ng.g¹ for aldrin, 0.2 - 1.84 ng.g¹ for dielddrin, 0.30 - 1.31 ng.g¹ for endrin, 0.11 - 0.26 ng.g¹ for lindane, 0.56- 3.36 ng.g¹ for heptachlor, 0.2- 1.51 ng.g¹ for P,P' DDE, 0.18- 2.91 ng.g¹ for P,P' DDD, 0.11 - 3.12 ng.g¹ for P,P' DDT. These results reveal that the sediments along coast of Chittagong are slightly contaminated with some of these organochlorine insecticides.

Glucocorticoid receptor and protein/RNA synthesis-dependent mechanisms underlie the control of synaptic plasticity by stress

Learning and memory are exquisitely sensitive to behavioral stress, but the underlying mechanisms are still poorly understood. Because activity-dependent persistent changes in synaptic strength are believed to mediate memory processes in brain areas such as the hippocampus we have examined the means by which stress affects synaptic plasticity in the CA1 region of the hippocampus of anesthetized rats, Inescapable behavioral stress (placement on an elevated platform for 30 min) switched the direction of plasticity, favoring low frequency stimulation-induced decreases in synaptic transmission (long-term depression, LTD), and opposing the induction of long-term potentiation by high frequency stimulation, We have discovered that glucocorticoid receptor activation mediates these effects of stress on LTD and longterm potentiation in a protein synthesis-dependent manner because they were prevented by the glucocorticoid receptor antagonist RU 38486 and the protein synthesis inhibitor emetine. Consistent with this, the ability of exogenously applied corticosterone in non-stressed rats to mimic the effects of stress on synaptic plasticity was also blocked by these agents, The enablement of low frequency stimulation-induced LTD by both stress and exogenous corticosterone was also blocked by the transcription inhibitor actinomycin D, Thus, naturally occurring synaptic plasticity is liable to be reversed in stressful situations via glucocorticoid receptor activation and mechanisms dependent on the synthesis of new protein and RNA, This indicates that the modulation of hippocampus-mediated learning by acute inescapable stress requires glucocorticoid receptor-dependent initiation of transcription and translation.

Morphine conditioned place preference depends on glucocorticoid receptors in both hippocampus and nucleus accumbens

Learned association between drugs of abuse and context is essential for the formation of drug conditioned place preference (CPP), which is believed to engage many brain regions including hippocampus, and nucleus accumbens (NAc). The underlying mechanisms

Survey of veterinery [sic] drug residues, environmental contaminants and mycotoxins of cultured shirmp [sic] in Iran

Economical achievement of optimal growth in developing countries may lead to sustainable poverty reduction. Agricultural activities play an important role in economy and human being welfare, which leads to establishment of food security and quality. Aquaculture products in developing countries share 51.4 percent of total agricultural production.7—percent in developed countries. Therefore undoutedly food production by means of quality and quantity has to be increased .The history of shirmp production goes back to 500 years ago. Today 50 countries of the world produce shirmp .In Islamic Republic of Iran shrimp production started since 1992 in the coastal region of Persian Gulf. The shrimp culture farms canbe classified in to 4 different categories; Extensive, semi-extensive, intensive and super instensive. Global ecological manitenanc is one of the major concerns of authorities Human manipulation of nature is the most destructive activity. Industrial sweage leakage in to the rivers and water sources is a big issue that causes reduction in the aquatic population. Heavy metals have an inhibitory effect in the production and growth of sealife. Human intake of food treated with anti microbial cause's allergy, hypersensitivity and develops microbial resistance. Organochlorine compounds contamination may found in hepato pancreatic tissues of aquatic products, Aresnic may transfer to man via plant & animal product contamination. In 1991 during Persian Gulf Mir 700 oil well set