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Subacute studies of monocrotophos [Dimethyl (E)-1-methyl-2-(methyl-carbamoyl) vinyl phosphate] on mosquito fish, Gambusia affinis, were carried out in vivo for 24 days to assess the locomotor behavior, structural integrity of gill, and targeted enzyme acetylcholinesterase (AChE, EC: 3.1.1.7) interactions. Monocrotophos (MCP) can be rated as moderately toxic to G. affinis, with a median lethal concentration (LC(50)) of 20.49 +/- 2.45 mgL(-1). The fish exposed to sublethal concentration of LC(10) (7.74 mgL(-1)) were under stress and altered their locomotor behavior, such as distance traveled per unit time (m min(-1)) and swimming speed (cm sec(-1)) with respect to the length of exposure. Inhibition in the activity of brain AChE and deformities in the primary and secondary lamellae of gill may have resulted in failure of exchange of gases. The maximum inhibition of 95% of AChE activity was observed on days 20 and 24. Morphological aberrations in the gills were also studied during exposure to the sublethal concentration of monocrotophos for a period ranging from 8 to 24 days. The extent of damage in gill was dependent on the duration of exposure. The findings revealed that inhibition in brain AChE activity and structural alteration in gill were responsible for altering the locomotor behavior of exposed fish.

The acute toxicity of azodrin (monocrotophos, an organophosphorus insecticide) was determined on a soil organism, Eisenia foetida. The median lethal concentrations (LC50) were derived from a 48-h paper contact test and from artificial soil tests. The LC50 of azodrin in the paper contact test was 0.46 +/- 0.1 microg cm(-2) (23 +/- 6 mg L(-1)) and those in the 7- and 14-day artificial soil tests were 171 +/- 21 and 132 +/- 20 mg kg(-1), respectively. The neurotoxic potentiality of azodrin was assessed by using a marker enzyme, acetylcholinesterase (AChE; EC 3.1.1.7) in both in vitro and in vivo experiments. The progressive signs of morphological destruction are correlated with percentage inhibition of AChE in the in vivo experiments. The kinetics of AChE activity in the presence and absence of azodrin indicated that the toxicant is competitive in nature. This study demonstrated that azodrin causes concentration-dependent changes in the morphology and AChE activity of the earthworm E. foetida.

Acute toxicity of monocrotophos (MCP) and its newly synthesized thiol analogs 2-butenoic acid-3-(diethoxyphosphinothionyl) methyl ester (RPR II) and 2-butenoic acid-3-(diethoxyphosphinothionyl) ethyl ester (RPR-V) was studied on euryhaline fish, Oreochromis mossambicus. The median lethal concentrations of MCP, RPR-II, and RPR-V are 11.506, 0.167, and 0.174 mgL(-1), respectively. Both the analogs were found to be 65-fold more toxic than MCP. Inhibitory and recovery patterns of brain, gill, and muscle acetylcholinesterase (AChE) was studied in vivo after exposure to LC(50) (96 h) concentrations. Recovery study was performed of regular intervals of day-1, -7, -14, -21, and -28 to establish the time course of 50% and 100% recovery in neurotransmitter enzyme of brain, gill, and muscle tissues. The sensitivity of the tissue AChE was in the order gill>brain>muscle. The relative toxicity of these compounds with regard to AChE was RPR-II>RPR-V>MCP. The MCP-treated fishes recovered at a faster rate than for RPR-V, followed by RPR-II.

Indian bullfrog Haplobatrachus tigerinus (Daudin) was exposed to sublethal dose (1/3 of LC50 I.E. 1.166 mg/kg) of fenvalerate technical grade and the effect was studied on the specific activity of acetyl cholinesterase in the different tissues of frog viz., brain, muscle, liver, kidney and testis at different time periods viz., 3,6, 12, 24, 48 and 72 hours. The inhibition of specific activity of acetyl cholinesterase was in the order of kidney > brain > muscle > liver > testis. A significant inhibition was noticed in kidney at 12 hours (-64.33%) and no effect was noticed at 3 hours in testis (+0.67%). The AChE activity was inhibited in first three hours of administration of fenvalerate in all the tissue tested. The inhibition continued upto 6 hours or 2 hours in different tissue but the recovery was started by 24 hours and almost completed by 72 hours.

The three-dimensional structures of human and rabbit liver cytosolic recombinant serine hydroxymethyltransferases (hcSHMT and rcSHMT) revealed that E75 and Y83 (numbering according to hcSHMT) are probable candidates for proton abstraction and Calpha-Cbeta bond cleavage in the reaction catalyzed by serine hydroxymethyltransferase. Both these residues are completely conserved in all serine hydroxymethyltransferases sequenced to date. In an attempt to decipher the role of these residues in sheep liver cytosolic recombinant serine hydroxymethyltransferase (scSHMT), E74 (corresponding residue is E75 in hcSHMT) was mutated to Q and K, and Y82 (corresponding residue is Y83 in hcSHMT) was mutated to F. The specific activities using serine as the substrate for the E74Q and E74K mutant enzymes were drastically reduced. These mutant enzymes catalyzed the transamination of D-alanine and 5,6,7, 8-tetrahydrofolate independent retroaldol cleavage of Lallo threonine at rates comparable with wild-type enzyme, suggesting that E74 was not involved directly in the proton abstraction step of catalysis, as predicted earlier from crystal structures of hcSHMT and rcSHMT. There was no change in the apparent Tm value of E74Q upon the addition of L-serine, whereas the apparent Tm value of scSHMT was enhanced by 10 degrees C. Differential scanning calorimetric data and proteolytic digestion patterns in the presence of L-serine showed that E74Q was different to scSHMT. These results indicated that E74 might be required for the conformational change involved in reaction specificity. It was predicted from the crystal structures of hcSHMT and rcSHMT that Y82 was involved in hemiacetal formation following Calpha-Cbeta bond cleavage of L-serine and mutation of this residue to F could lead to a rapid release of HCHO. However, the Y82F mutant had only 5% of the activity and failed to form a quinonoid intermediate, suggesting that this residue is not involved in the formation of the hemiacetal intermediate, but might be involved indirectly in the abstraction of the proton and in stabilizing the quinonoid intermediate.

Serine hydroxymethyltransferase belongs to the alpha class of pyridoxal-5'-phosphate enzymes along with aspartate aminotransferase. Recent reports on the three-dimensional structure of human liver cytosolic serine hydroxymethyltransferase had suggested a high degree of similarity between the active-site geometries of the two enzymes. A comparison of the sequences of serine hydroxymethyltransferases revealed the presence of several highly conserved residues, including Pro-297. This residue is equivalent to residue Arg-292 of aspartate aminotransferase, which binds the gamma-carboxy group of aspartate. In an attempt to change the reaction specificity of the hydroxymethyltransferase to that of an aminotransferase and to assign a possible reason for the conserved nature of Pro-297, it was mutated to Arg. The mutation decreased the hydroxymethyltransferase activity significantly (by 85-90%) and abolished the ability to catalyse alternative reactions, without alteration in the oligomeric structure, pyridoxal 5'-phosphate content or substrate binding. However, the concentration of the quinonoid intermediate and the extent of proton exchange was decreased considerably (by approx. 85%) corresponding to the decrease in catalytic activity. Interestingly, mutant Pro-297 Arg was unable to perform the transamination reaction with L-aspartate. All these results suggest that although Pro-297 is indirectly involved in catalysis, it might not have any role in imparting substrate specificity, unlike the similarly positioned Arg-292 in aspartate aminotransferase.

Phytochemical investigation on the constituents of heartwood of Pterocarpus santalinus resulted in the isolation of a new acylated isoflavone glucoside. The structure of the new compound was elucidated on the basis of spectral studies as 4',5-dihydroxy-7-O-methyl isoflavone 3'-O-D-(3''-E-cinnamoyl)glucoside.

Bud break and multiple shoots were induced in apical and axillary meristems derived from one month old seedlings of S. mukorossi on Murashige and Skoog (MS) medium supplemented with benzylamino purine (BAP) 0.4 microM or 0.8 microM alone. A combination of BAP and gibberellic acid (GA3) 0.4 microM and 2.8 microM produced elongated multiple shoots from both types of explants. Excised shoots were rooted on MS medium respectively with indole-3-butyric acid (IBA) 3.4 microM or 2.4 microM. The regenerated plantlets were successfully acclimatized and transferred to soil.

A new isoflavone together with liquiritigenin and isoliquiritigenin has been isolated from the heartwood of Pterocarpus santalinus. Based on spectral methods, the structure of the new compound was elucidated as 6-hydroxy,7,2',4',5'-tetramethoxyisoflavone.

A new pentacyclic triterpene was isolated from the callus induced from the stem cuttings of Pterocarpus santalinus. Based on spectral methods, the structure of the new compound was elucidated as 3-ketooleanane (1).

A new isoflavone glucoside (1) together with the known santal has been isolated from the heartwood of Pterocarpus santalinus. Based on spectral methods, the structure of the new compound was elucidated as 4',5-dihydroxy 7-O-methyl isoflavone 3'-O-beta-D-glucoside.

Aspartate residues function as proton acceptors in catalysis and are involved in ionic interactions stabilizing subunit assembly. In an attempt to unravel the role of a conserved aspartate (D89) in sheep-liver tetrameric serine hydroxymethyltransferase (SHMT), it was converted into aspargine by site-directed mutagenesis. The purified D89N mutant enzyme had a lower specific activity compared with the wild-type enzyme. It was a mixture of dimers and tetramers with the proportion of tetramers increasing with an increase in the pyridoxal-5'-phosphate (PLP) concentration used during purification. The D89N mutant tetramer was as active as the wild-type enzyme and had similar kinetic and spectral properties in the presence of 500 microM PLP. The quinonoid spectral intermediate commonly seen in the case of SHMT was also seen in the case of D89N mutant tetramer, although the amount of intermediate formed was lower. Although the purified dimer exhibited visible absorbance at 425 nm, it had a negligible visible CD spectrum at 425 nm and was only 5% active. The apo-D89N mutant tetramer was a dimer unlike the apo-form of the wild-type enzyme which was present predominantly as a tetramer. Furthermore the apo mutant dimer could not be reconstituted to the holo-form by the addition of excess PLP, suggesting that dimer-dimer interactions are weak in this mutant. The recently published crystal structure of human liver cytosolic recombinant SHMT indicates that this residue (D90 in the human enzyme) is located at the N-terminal end of the fourth helix of one subunit and packs against K39 from the second N-terminal helix of the other symmetry related subunit forming the tight dimer. D89 is at the interface of tight dimers where the PLP 5'-phosphate is also bound. Mutation of D89 could lead to weakened ionic interactions in the tight dimer interface, resulting in decreased affinity of the enzyme for the cofactor.

Some organotin(IV) and tin(II) complexes of composition R(3)Sn[R'COC:CON(C(6)H(5))N:CCH(3)] (where R=C(4)H(9), R'=CH(3), C(3)H(5), p-ClC(6)H(4); R=C(6)H(5), R'=C(6)H(5) and p-ClC(6)H(4)) and Sn[p-ClC(6)H(4)COC:CON(C(6)H(5))N:CCH(3)](2) were screened for their toxicity against Aedes aegypti larvae. Organotin(IV) complexes were more active than tin(II) complexes.

Sequence analysis and metal ion binding studies reported earlier have established that the calcium binding protein (CaBP) from the parasitic ameboid Entamoeba histolytica protein has four canonical EF hand motifs which bind calcium. Equilibrium denaturation studies on both the apo and the holo forms of this protein indicate the presence of stable transition intermediates at low denaturant concentrations as revealed by the binding of the non-specific hydrophobic dye ANS. Fast reaction kinetics shows that the binding of the Gdn+ ions at or near the Ca2+ sites in the N-terminal domain influences metal ion binding to the sites in the C-terminal domain. Isothermal calorimetric titrations performed using low GdnHCl concentrations reveal the presence of two binding sites of low affinity, both being endothermic in nature. Thus the stabilization of CaBP observed at low GdnHCl concentration represents a native-like intermediate, with the Gdn+ ions mimicking Ca2+ binding at the N-terminal domain of this protein.

In vitro effects of six dibenzyltin dichloride (A-series) and another six dibenzyltin diisothiocyanate (B-series) complexes on Na+/K+-ATPase were studied. The rat brain synaptic membranes were prepared and significant ATPase inhibition was observed with all the complexes in a dose-dependent manner. The order of potency on Na+/K+-ATPase activity of the chloride complexes was found to be higher than that of the isothiocyanate complexes. The IC50 values range from 64 microM to 252 microM for the chloride complexes and from 132 microM to 399 microM for the isothiocyanate complexes. A structure-activity relationship could be derived for the chloride complexes.

Acetylcholinesterase (AChE, EC 3.1.1.7) was purified from western corn rootworm (WCR, Diabrotica virgifera virgifera) beetles by affinity chromatography. The purification factor reached over 20,000-fold with a specific activity of 169.5 mumol/min/mg and a yield of 23%. The Vmax values for hydrolyzing acetylthiocholine (ATC), acetyl-(beta-methyl) thiocholine (A beta MTC), propionylthiocholine (PTC), and S-butyrylthiocholine (BTC) were 184.8, 140.5, 150.2, and 18.8 mumol/min/mg, respectively, and K(m) values were 19.7, 18.5, 14.1, and 11.0 microM, respectively. The first three substrates showed significant inhibition to the AChE at higher concentrations, whereas BTC showed inhibition at the concentrations of 0.25-2 nM but activation at > 4 mM. AChE activity was almost completely inhibited by 1 microM eserine and BW284C15, respectively, but only 12% of AChE activity were inhibited by ethopropazine at the same concentration. These results suggested that the purified AChE from WCR was a typical insect AChE. Insecticides or their oxidative metabolites, chlorpyrifos-methyl oxon, carbofuran, carbaryl, malaoxon, and paraoxon, used in in vitro kinetic study exhibited high inhibition to AChE purified from WCR. However, chlorpyrifos-methyl oxon and carbofuran showed at least 36- and 4-fold, respectively, higher inhibitory potency than the remaining insecticides examined. Results from our in vitro inhibition of AChE agreed quite well with the previously published in vivo bioassay data.

Alkali extract of sepia shell possesses hypoglycemic effect. The status of glycogen and pyruvate and the activity of glucose-6-phosphatase and alanine amino transferase in liver was studied under the influence of sepia shell extract in both normal and streptozotocin induced diabetic mice. The glycogen concentration was elevated steeply in both and the pyruvate concentration increased substantially in diabetic mice, while the activity of glucose-6-phosphatase and alanine amino transferase was inhibited in normal and diabetic mice. The sepia shell extract enhances glycogenesis and reduces the formation of glucose from metabolic intermediates like pyruvate and glucose-1-phosphate and by suppressing gluconeogenesis.

Species-related differences in sensitivity to acute intoxication by anticholinesterase (anti-ChE) compounds have been attributed, in large part, to differences in the kinetics of inhibition of acetylcholinesterase (AChE: EC 3.1.1.7) in vitro. The following investigation was designed to determine if species-related differences in the sensitivity of brain AChE to inhibition by monocrotophos (MCP) could contribute to the interspecies differences in toxicity. Brain AChE activity was significantly greater in fish followed by pigeon and rat. MCP was found to be a competitive inhibitor of rat, pigeon, and fish brain AChE, thereby, altering the Km (Michaelis constant) widely among the species. Comparatively, least alterations in Km were observed in fish and maximum in pigeon. The Ki (bimolecular inhibition constant) of rat was 1.4- and 3.2-fold lower than that of pigeon and fish, respectively. Although fish brain had significantly greater AChE activity, it was the least sensitive to MCP inhibition. These data suggest that the greater sensitivity of rodent brain AChE to inhibition by MCP may contribute to the greater toxicity of MCP in rodents than in birds and fishes.

Two simple and sensitive visible spectrophotometric methods (methods A and B) for the determination of tolnaftate in bulk samples and formulations are described. Method A (lambda(max) 490 nm) involves the reaction of tolnaftate with 2,6-dichloroquinone-4-chlorimide (DCQC: Gibb's reagent), while method B (lambda(max) 530 nm) is based on a similar reaction of the drug with p-N,N-dimethylphenylene diamine in the presence of chloramine-T. Both these methods are applicable to pure samples as well as formulations of the drug.

Fertile White Leghorn chicken eggs were injected on day 4 of incubation with 0, 0.25, 0.50 or 1.00 mg RPR-V/egg and opened on day 20 or allowed to hatch. The following parameters were examined: hatchability, growth and development, external malformations and skeletal deformities. As the dose was increased, the hatchability decreased, and the incidence of deformities increased. Staining of the skeleton with Alizarin red showed clear deformities in the embryos. The results suggest that RPR-V has teratogenic effects on chick embryos when injected on day 4 of incubation.

Chronopharmacokinetics of rifampicin was studied in four healthy adult male human volunteers after drug (2.0 g) ingestion at 6.00, 12.00, 18.00 and 24.00 hr. The absorption rate constant was found to be lower and the time to reach peak concentration was longer after drug administration at 24.00 hr than at other dosing times. A second peak was observed in all individual volunteers between 6-12 hr after drug dosing at 24.00 hr. This may be due to the influence of biliary rhythms on the disposition kinetics of rifampicin.

Exposure of a freshwater fish to several sublethal concentrations (1.25, 2.50, 5.00, 10.00, and 20.00 mg/liter) of lead for a period of 30 days showed significant accumulation of lead in the blood and tissues. Lead bioaccumulated in the study showed organ-specific distribution, with high levels in the blood followed by the kidney, gill, liver, and brain and comparatively lesser amounts in the ovary and muscle tissues. The lead accumulation in tissues was found to increase with lead in water up to a concentration of 5 mg/liter, and at concentrations of 10 and 20 mg/liter the lead accumulation in the tissues, although indicating an increase, was not proportional to the concentration in water. Exposure of the freshwater fish Anabas testudineus to a sublethal (5 ppm) concentration of lead nitrate for a period of 30 days during the preparatory phase of its annual reproductive cycle reduced the total lipids, phospholipids, and cholesterol levels in the liver and ovary tissues while the free fatty acid levels were increased and lipase activity was elevated. All the parameters in the blood were found to increase. These results suggest that lead nitrate affects the lipid metabolism of the fish and this may reduce the fecundity of the fish since lipids are known to play an important role in teleost reproduction as an energy source and a precursor of steroids.

A simple, selective and sensitive spectrophotometric method has been developed for the determination of microgram quantities of warfarin sodium (WS), nicoumalone (NIC) and acebutolol hydrochloride (ACBH), either in pure form or in pharmaceutical preparations. This method is based on the haloform reaction with a known and excess of standard iodine solution under alkaline conditions. The excess of iodine is determined at pH 3.0 with metol-INH. The absorbance of the resulting p-N-methyl-benzoquinonemonoimine-INH charge-transfer complex is measured at 620 nm.

The strain differences in the neurotoxic potential of monocrotophos (MCP) were assessed by determining the inhibition of brain acetylcholinesterase (AChE) in BALB/cAnN, DBA/2J and C57BL/6J in vitro. MCP being a competitive inhibitor for AChE, alters the Km values widely among these inbred strains. Comparatively least alterations in Km were found in BALB/cAnN and maximum in DBA/2J. Based on the Ki values DBA/2J was found to be the most sensitive strain to MCP inhibition followed by C57BL/6J and BALB/cAnN.

The effect of 16 L-amino acids on the activity levels of arginase in sheep brain homogenates was studied. The amino acids leucine, valine, lysine, and ornithine inhibited arginase activity significantly. The other amino acids tested did not show a significant influence on arginase activity. The inhibition was related to the carbon chain length of the amino acids.

An experiment for studying the effect of lime-pelleting of inoculated seed of soybean [Glycine max. (L.) Merr.] on nodulation, growth, and grain yield was undertaken in red sandy loam of Bangalore, having a pH of 4.0. The results indicated that inoculation alone increased significantly nodulation, plant dry weight, and grain yield. But inoculation plus lime-pelleting significantly increased the dry weight of nodules and plants, but not the grain yield. However, lime-pelleting was found to be beneficial though not essential in acid soils.

Exposure of fresh water crab Barytelphusa querini to a sublethal concentration of NaF (30 ppm) caused significant alterations in the carbohydrate metabolism. In an exposure span of 15 days the crab was observed to have marked depletion in glycogen and total free sugar levels. A significant elevation in "active" and "total" glycogen phosphorylase activity was observed. The activity changes of lactate dehydrogenase reflected changes in tissue lactate levels and succinate dehydrogenase activity had a tissue specificity.

Respiration and related physiologic variables in different tissues of Barytelphusa guerini and the respiration of this freshwater crab as a whole are closely synchronized in phase and in frequency along the circadian scale, in the face of large differences in circadian amplitude. A very close timing of most of the 36 variables examined in 2 separate circadian profiles and a modulation of some of these variables in added profiles as a function of lunar stage are clearly demonstrable, statistically significant and illustrative of time relations at 2 interacting frequencies.