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.     

Methyl mercury uptake by free and immobilized cyanobacterium

Methyl mercury uptake in free cells and different immobilizates of the cyanobacteriumNostoc calcicola has been examined. The general growth of the immobilized cyanobacterial cells could be negatively correlated with methyl mercury uptake. Alginate spheres proved most efficient in terms of uptake rate (0.48 nmol mg protein^–1 min^–1, 10 min) and total bioaccumulation (10.71 nmol mg protein^–1, 1 h) with a bioconcentration factor of 3.3×10³. Alginate biofilms showed a faster methyl mercury accumulation rate (0.83 nmol mg protein^–1 min^–1, 10 min) with a saturation of 10.28 nmol mg protein^–1 reached within only 30 min (bioconcentration factor, 3.1×10³). Foam preparations with a slow initial uptake approximated biofilms but were characterized by a lower bioconcentration factor (2.8×10³). Free cells, in comparison, maintained the initial slow rate of uptake (0.62 nmol mg protein^–1 min^–1, 10 min), saturating at 30 min (8.81 nmol mg protein^–1), and the resultant lowest bioconcentration factor (2.7×10³). Cell ageing (30 days) brought a drastic reduction (3-fold) in organomercury uptake by free cells while alginate spheres maintained the same potential. Foam preparations of the same age showed a significant improvement in methyl mercury uptake followed by only a marginal decline in alginate biofilms. Data are discussed in the light of the physiological efficiency and longevity of immobilized cells.     

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.

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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.

     

In vitro Production of Halo Blight Inducing Toxin by Pseudomonas phaseolicola (Burkh.) Dowson

Three pathogenic strains of Pseudomonas phaseolicola (strain 1 and 3 virulent and strain 5 weakly virulent) were tested for their toxic activity. All three strains produced detectable amounts of toxin in vitro. Cultural conditions and length of incubation greatly influenced toxin production. Maximum amount of toxin was produced at 20°C and pH 6.5. Glycerol served as the best carbon source and 1-cysteine as the best amino acid for toxin production.     

Intergeneric protoplast fusion between Acinetobacter sp. A3 and Pseudomonas putida DP99 for enchanced hydrocarbon degradation

Summary Polyethylene glycol 6000 mediated protoplast fusion between an alkane degrader Acinetobacter sp. A3, and a naphthalene degrader, Pseudomonas putida DP99 , resulted in fusants capable of degrading both hydrocarbons and were morphologically similar to Acinetobacter sp. A3. While fusant F4/13 and Pseudomonas putida DP99 degraded over 98% of naphthalene provided by the end of five days, tetradecane degradation by fusant F4/13 was 82% compared to 77% by Acinetobacter sp. A3 in the same time period. Also, while from naphthalene +tetradecane mixture, fusant F4/13 could degrade 99% and 53% of naphthalene and tetradecane respectively, both the parent strains together could degrade over 99% naphthalene but only about 16% tetradecane.     

Collagenase-3 (MMP-13) deficiency protects C57BL/6 mice from antibody-induced arthritis

Introduction

Matrix metalloproteinases (MMPs) are important in tissue remodelling. Here we investigate the role of collagenase-3 (MMP-13) in antibody-induced arthritis.

Methods

For this study we employed the K/BxN serum-induced arthritis model. Arthritis was induced in C57BL/6 wild type (WT) and MMP-13-deficient (MMP-13^–/–) mice by intraperitoneal injection of 200 μl of K/BxN serum. Arthritis was assessed by measuring the ankle swelling. During the course of the experiments, mice were sacrificed every second day for histological examination of the ankle joints. Ankle sections were evaluated histologically for infiltration of inflammatory cells, pannus tissue formation and bone/cartilage destruction. Semi-quantitative PCR was used to determine MMP-13 expression levels in ankle joints of untreated and K/BxN serum-injected mice.

Results

This study shows that MMP-13 is a regulator of inflammation. We observed increased expression of MMP-13 in ankle joints of WT mice during K/BxN serum-induced arthritis and both K/BxN serum-treated WT and MMP-13^–/– mice developed progressive arthritis with a similar onset. However, MMP-13^–/– mice showed significantly reduced disease over the whole arthritic period. Ankle joints of WT mice showed severe joint destruction with extensive inflammation and erosion of cartilage and bone. In contrast, MMP-13^–/– mice displayed significantly decreased severity of arthritis (50% to 60%) as analyzed by clinical and histological scoring methods.

Conclusions

MMP-13 deficiency acts to suppress the local inflammatory responses. Therefore, MMP-13 has a role in the pathogenesis of arthritis, suggesting MMP-13 is a potential therapeutic target.     

The Small GTPase RhoA Is Required for Proper Locomotor Circuit Assembly

The assembly of neuronal circuits during development requires the precise navigation of axons, which is controlled by attractive and repulsive guidance cues. In the developing spinal cord, ephrinB3 functions as a short-range repulsive cue that prevents EphA4 receptor-expressing corticospinal tract and spinal interneuron axons from crossing the midline, ensuring proper formation of locomotor circuits. Here we report that the small GTPase RhoA, a key regulator of cytoskeletal dynamics, is also required for ephrinB3/EphA4-dependent locomotor circuit formation. Deletion of RhoA from neural progenitor cells results in mice that exhibit a rabbit-like hopping gait, which phenocopies mice lacking ephrinB3 or EphA4. Consistent with this locomotor defect, we found that corticospinal tract axons and spinal interneuron projections from RhoA-deficient mice aberrantly cross the spinal cord midline. Furthermore, we determined that loss of RhoA blocks ephrinB3-induced growth cone collapse of cortical axons and disrupts ephrinB3 expression at the spinal cord midline. Collectively, our results demonstrate that RhoA is essential for the ephrinB3/EphA4-dependent assembly of cortical and spinal motor circuits that control normal locomotor behavior.