Identification of an enzymatic activity that hydrolyzes protein-bound ADP-ribose in skeletal muscle

An enzymatic activity present in high-speed supernatant fluids of rat skeletal muscle was found that catalyzes the release of ADP-ribose from ADP-ribosylated-modified lysozyme. The nature of the product was proved by chromatographic studies and proton nuclear magnetic resonance spectroscopy. The enzyme activity is stimulated by Mg2+, dithioerythritol, and flouride. These results and those published earlier (Soman, G., Mickelson, J.R., Louis, C.F., and Graves, D.J. (1984) Biochem. Biophys. Res. Commun. 120, 973-980) show that ADP-ribosylation is a reversible process in skeletal muscle.     

Endogenous ADP-ribosylation in skeletal muscle membranes

The characteristics of ADP-ribosyltransferase activity in skeletal muscle membranes have been studied. The membrane enzymes can ADP-ribosylate exogenous substrates such as guanylhydrazones, polyarginine, lysozyme, and histones. The properties of the enzyme are investigated by using diethylaminobenzylidineaminoguanidine as a model substrate. Incubation of the membranes with [32P]adenylate-labeled NAD results in the labeling of a number of cellular proteins. Magnesium ions, detergents, and diethylaminobenzylidineaminoguanidine stimulated the ADP-ribosylation of membrane proteins, whereas L-arginine methyl ester and arginine inhibited ADP-ribosylation. The labeling of specific proteins in the sarcoplasmic reticulum and glycogen pellet is influenced significantly by detergents, nucleotides, and thiols. The hydroxylamine sensitivity of the ADP-ribose linkage in the membrane proteins is similar to that reported for (ADP-ribose)-arginine linkage. Snake venom phosphodiesterase digestion of the ADP-ribosylated membranes produces 5'-AMP as the major acid-soluble digestion product. The results suggest that the primary mode of modification is mono(ADP-ribosyl)ation. The ADP-ribosyltransferase activity in the membrane preparations is not extracted under conditions used for solubilization of extrinsic proteins, suggesting that the activity is associated with some integral membrane protein.     

Local Perceptions of Federal Block Grants

We report the results of a survey of California county medical societies and county health officers on federal health block grants. Survey respondents agreed that the existing network of health services funded through the federal block grants and the current statelocal apparatus for providing these services are sound. Most respondents do not recommend major changes in the service system, and most support a strong state role in administering programs under the block grants.     

The role of leaf surface wetness in larval behaviour of the sorghum shoot fly, Atherigona soccata

The susceptibility of sorghum to the shoot fly Atherigona soccata Rondani, (Diptera: Muscidae) is affected by seedling age and is highest when seedlings are 8–12 days old. This corresponds with high moisture accumulation on the central leaf which is the path of newly hatched larva as it moves downwards from the oviposition site, towards the growing apex. Studies showed that leaf surface wetness (LSW) of the central shoot leaf was higher in 10-day old seedlings than in seedlings of other ages. Similarly, LSW was much higher in the susceptible sorghum genotype CSH 1 than in the resistant genotype IS 2146. Larvae moved faster towards the growing point and produced deadhearts much earlier in CSH 1 than in IS 2146. They also moved faster in 10-day old seedlings than in seedlings of other ages. It was also shown that the leaf surface wetness of the central shoot leaf is a more reliable parameter of resistance than the glossy leaf trait or trichome density.

---

L'influence de la humidité de la surface foliaire sur le comportement de la mouche des pousses du sorgho

Résumé La sensibilité du sorgho à la mouche des pousses du sorgho, Atherigona soccata Rondani, est liée à l'âge de la plantule. Elle est plus forte lorsque la plantule est âgée de 8 à 12 jours et la sensibilité est maximale à 10 jours. A ce stade de croissance on observe une forte accumulation d'humidité sur la feuille centrale de la tige. Les jeunes larves traversent cette zone humide lorsqu'elles descendent vers la zone de croissance à partir des pontes déposées sur la face ventrale des feuilles déroulées.Des études ont été menées à l'ICRISAT (Inde) sur la relation entre l'humidité de la feuille centrale de la tige des plantules du sorgho et les dégâts provoqués par la mouche des pousses. L'humidité de la surface des feuilles (HSF) a été estimée grâce à une échelle visuelle graduée 1 à 5 où, 1 = pas d'humidité apparente et 5 = surface de la feuille recouverte de gouttes d'eau. La HSF est plus élevée sur des pousses de sorgho âgées de 10 j que sur les pousses appartenant à d'autres classes d'âge. Les valeurs observées sont également plus fortes pour les variétés non résistantes à ce ravageur (CSH 1,4.8) que pour les variétés résistantes (IS 2146, (2)). La vitesse du déplacement larvaire entre le cornet et la zone de la croissance varie en fonction de l'âge de la plante et des cultivars. Les larves migrent plus rapidement vers la zone de croissance et provoquent la mort du coeur du sorgho plus tôt dans la variété CSH 1 que dans IS 2146. Les larves se déplacent plus rapidement dans les pousses âgées de 10 j que dans les pousses appartenant à d'autres classes d'âge.Des études ont également démontré que la HSF n'est pas directement liée au caractère feuille lisse où à la densité des trichomes. La HSF est faible pour les génotypes résistants présentent où non le caractère feuille lisse. Par contre la HSF est élevée pour les génotypes non résistants présentant le caractère feuille lisse ou non. Aucune relation directe entre la densité des trichomes et les dégâts provoqués par la mouche des pousses n'a pu être mise en évidence. L'analyse des correlations établie pour les caractères de surface des feuilles avec la mort du cur des sorghos indique que les correlations sont faibles et non-significatives pour le caractère feuille lisse (0.49) et la densité des trichomes (0.39 et 0.2). Par contre les correlations sont fortes et significatives pour la HSF (0.82).On conclue que la HSF de la feuille centrale de la tige est un facteur important dans le déterminisme de la résistance du sorgho vis à vis de la mouche des pousses. Les relations entre les processus physiologiques de la plante et les facteurs impliquées dans l'accumulation d'eau sur la surface des feuilles font actuellement l'objet d'études détaillées.

     

Deregulation of hepatic lipid metabolism associated with insulin resistance in rats subjected to chronic monocrotophos exposure

In our previous study, we demonstrated the potential of monocrotophos (MCP), an organophosphorus insecticide (OPI), to induce glucose intolerance, insulin resistance (IR), and dyslipidemia with hyperinsulinemia in rats after chronic exposure. As hyperinsulinemia is likely to exert an impact on hepatic lipid metabolism, we carried out this study to establish the effect of chronic MCP exposure (0.9 and 1.8 mg/kg/day for 180 days) on hepatic lipid metabolism in rats. The state of IR induced by MCP in rats was associated with an increase in the liver lipid content (triglyceride and cholesterol) and expression levels of sterol regulatory element‐binding proteins, PPARγ, acetyl‐CoA carboxylase, and fatty acid synthase in the liver. Similarly, activities of key enzymes (acetyl‐COA carboxylase, fatty acid synthase, lipin 1, malic enzyme, glucose‐6‐phosphate dehydrogenase, and glycerol‐3‐phosphate dehydrogenase), which regulate lipogenesis, were enhanced in livers of pesticide‐treated rats. A strong correlation was observed between insulin levels, hepatic lipid content, and plasma lipid profile in treated rats. Our study suggests that long‐term exposure to OPIs not only has a propensity to induce a state of hyperinsulinemic IR, but it is also associated with augmented hepatic lipogenesis, which may explain dyslipidemia induced by chronic exposure to MCP.     

Novel method to differentiate 3T3 L1 cells in vitro to produce highly sensitive adipocytes for a GLUT4 mediated glucose uptake using fluorescent glucose analog

Adipocytes play a vital role in glucose metabolism. 3T3 L1 pre adipocytes after differentiation to adipocytes serve as excellent in vitro models and are useful tools in understanding the glucose metabolism. The traditional approaches adopted in pre adipocyte differentiation are lengthy exercises involving the usage of IBMX and Dexamethasone. Any effort to shorten the time of differentiation and quality expression of functional differentiation in 3T3 L1 cells in terms of enhanced Insulin sensitivity has an advantage in the drug discovery process. Thus, there is a need to develop a new effective method of differentiating the pre adipocytes to adipocytes and to use such methods for developing efficacious therapeutic molecules. We observed that a combination of Dexamethasone and Troglitazone generated differentiated adipocytes over fewer days as compared to the combination of IBMX and Dexamethasone which constitutes the standard protocol followed in our laboratory. The experiments conducted to compare the quality of differentiation yielded by various differentiating agents indicated that the lipid droplet accumulation increased by 112 % and the GLUT4 mediated glucose uptake by 137 % in cells differentiated with Troglitazone and Dexamethasone than in cells differentiated traditionally. The comparative studies conducted for evaluating efficient measurable glucose uptake by GOPOD assay, radioactive ³H-2-deoxy-D-glucose assay and by non-radioactive 6-NBDG (fluorescent analog of glucose) indicated that the non-radioactive method using 6-NBDG showed a higher signal to noise ratio than the conventional indirect glucose uptake method (GOPOD assay) and the radioactive ³H-2-deoxy-D-glucose uptake method. Differentiated 3T3 L1 cells when triggered with 2.5 ng/mL of Insulin showed 3.3 fold more glucose uptake in non-radioactive method over the radioactive ³H-2-deoxy-D-glucose uptake method. The results of this study have suggested that a combination of Dexamethasone and Troglitazone for 3T3 L1 cell differentiation helps in better quality differentiation over a short period of time with increased sensitivity to Insulin. The application of these findings for developing new methods of screening novel Insulin mimetics and for evaluating the immunological responses has been discussed.     

Identification of Penicillin-binding proteins employing support vector machines and random forest

Penicillin-Binding Proteins are peptidases that play an important role in cell-wall biogenesis in bacteria and thus maintaining bacterial infections. A wide class of β-lactam drugs are known to act on these proteins and inhibit bacterial infections by disrupting the cell-wall biogenesis pathway. Penicillin-Binding proteins have recently gained importance with the increase in the number of multi-drug resistant bacteria. In this work, we have collected a dataset of over 700 Penicillin-Binding and non-Penicillin Binding Proteins and extracted various sequence-related features. We then created models to classify the proteins into Penicillin-Binding and non-binding using supervised machine learning algorithms such as Support Vector Machines and Random Forest. We obtain a good classification performance for both the models using both the methods.     

InDiaMed: A Comprehensive Database of Indian Medicinal plants for Diabetes

According to International Diabetes Federation (IDF), India has 62.4 million people with diabetes and by 2030 it is predicted that the number will rise to 100 million. Studies claim that there are around 410 experimentally proven Indian medicinal plants which have anti-diabetic activity, of which the mechanism of action of 109 plants has been elucidated or reported. So, the need of the hour is to explore the claims of Indian medicinal flora and open up the facets of many Indian plants which are being examined for their beneficial role in diabetes. So, we created a database (InDiaMed) of Indian medicinal plants that captures their role in anti-diabetic activity. InDiaMed''s features include chemical, pharmacological, biochemical and geographical information of the medicinal plant, scientifically relevant information of the plant, and the coherent research done on it in the field of diabetes. The database also includes the list of poly-herbal formulations which are used for treatment of diabetes in India.

Availability

http://www.indiamed.info