Susceptibility of testicular cell cultures of crab, Scylla serrata (Forskal) to white spot syndrome virus.

Testicular cell culture of crab, Scylla serrata (Forskal) was used to study the effects of White spot syndrome virus (WSSV). We are showing the susceptibility of cell culture of crabs to WSSV. The proliferating cell culture of testes were maintained for more than 4 months in a medium prepared from L15 and crab saline supplemented with epidermal growth factor. The cell cultures inoculated with different concentrations of virus showed distinct cytopathic effects such as change in cell appearance, shrinkage and cell lysis. WSSV infection of cultured cells was confirmed by Nested PCR technique. The incorporation of viral DNA in cultured cells was shown by RAPD profile generated using 10-mer primers. The controls that were not exposed to WSSV did not show cytopathic effects. This work shows the usefulness of proliferating testicular cell culture for studying WSSV infection using molecular tools. Thus, this report gains significance as it opens new vistas for diagnostics and drugs for WSSV.

Development of cell line from the testicular tissues of crab Scylla serrata.

This is the first report on development of a finite cell line from testicular tissues of crab, Scylla serrata. Both the explant and segregated tissues of testes yielded cells that could proliferate and grow. These cells ranged in size from 10 to 38 µm with distinct nuclei of varying shapes. The testicular cells survived and proliferated best in L-15-crab saline medium supplemented with epidermal growth factor (20 ng/mL) and glucose (1 mg/mL). The cell proliferation rate was assessed by Methyl tetrazolium assay in terms of change in optical density which clearly indicated a prominent increase in cell density. The testicular cells were subcultured at an interval of 4-6 days. These subcultured cells remained healthy and proliferated for 5 months with a minimum of ten subsequent passages. The finite cell line was characterized in terms of morphology, growth rate, lactate dehydrogenase release (for detecting health status) and 18S rRNA sequencing. This cell line could be a very useful tool for testing infections and replications of crustacean viruses. The present work provides a technique that could be extended for developing other crustacean cell lines.

Study of epileptiform activity in cerebral ganglion of mud crab Scylla serrata.

An attempt is made to induce in mud crab (Scylla serrata) epileptiform activities that resemble the generalized epileptic seizures. Cerebral ganglion of crab was exposed in situ, to a convulsant drug pentylenetetrazole (PTZ) 100 mM, for induction of seizures. Also, crabs were pretreated with antiepileptic drug viz sodium valproate (120 µmol/l) to inhibit epileptiform activities. The surface electrical discharges of cerebral ganglion were recorded using Unkelscope (MIT, USA) in control as well as experimental animals. The cerebral ganglion of crab showed a pattern of high cerebral electrical discharges after PTZ treatment compared to control. The sodium valproate promoted sedative action in control and prevented PTZ-mediated epileptiform discharges. Glutamate and GABA contents in cerebral ganglion were assayed. Glutamate level increased (31.45%) during PTZ treatment with concomitant decrease (43.93%) in GABA. Sodium valproate had no effect on glutamate concentration, but it decreased GABA by 24.75%. The present study shows that epileptiform activities can be induced in crabs.

Development of primary cell culture from Scylla serrata : Primary cell cultures from Scylla serrata.

This paper reports for the first time, the Primary cell culture of hepatopancreas from edible crab Scylla serrata using crab saline, L-15 (Leibovitz), 1 x L-15 + crab saline, 2 x L-15 + crab saline, 3 x L-15 and citrate buffer without any serum. We could isolate and maintain E (Embryonalzellen), F (Fibrenzellen), B (Blasenzellen), R (Restzellen) and G (Granular cells). Upon seeding the hepatopancreatic E, F, B, and R cells showed different survival pattern over time than granular cells. A modified L-15 (3x) medium supported the best survival of hepatopancreatic E, F B, and R cells in in-vitro culture. However granular cells could be maintained for 184 days with L-15 (1x) + crab saline. Fetal bovine serum was not effective additive and hampered cell viability in present study.

The study of Na+, K(+)-ATPase activity of rat brain during Crush syndrome.

Crush syndrome (CS) results from severe traumatic damage to the organism that is characterized by stress, acute homeostatic failure of the tissues, and myoglobinuria with severe intoxication. This leads to an acute impairment of kidneys and heart. The peripheral and central nervous systems are also the subject of significant changes in CS. Na(+), K(+)-ATPase is a critical enzyme in neuron that is essential for the regulation of neuronal membrane potential, cell volume as well as transmembrane fluxes of Ca(++) and Excitatory Amino Acids. In the present study, Na(+), K(+)-ATPase activity of rat brain regions [Olfactory lobes (OL), Cerebral cortex (CC), Cerebellum (CL), and Medulla oblongata (MO)] during CS was investigated. Experimental model of CS in albino rats was induced by 2-h of compression followed by 2, 24, and 48-h of decompression of femoral muscle tissue. In this study, we have observed elevation in Na(+), K(+)-ATPase activity above normal/control levels in all parts of brain (OL: 34.4%; CC: 1.0%; CL: 3.3% and MO: 45%) during 2-h compression in comparison to controls.

Structural characterization of vivapain-2 and vivapain-3, cysteine proteases from Plasmodium vivax: comparative protein modeling and docking studies.

Malaria remains one of the most important infectious diseases in the world. Plasmodial cysteine proteases are proposed to be promising targets for novel antimalarial drug design. Vivapain-2 and vivapain-3 are cysteine proteases from Plasmodium vivax and apparent orthologs of falcipain-2 and falcipain-3 from Plasmodium falciparum. Model structures of vivapain-2 and vivapain-3 have been derived using the comparative protein modeling approach and validated by various structure/geometry verification tools. Correlation between the interaction energies calculated based on the docking studies of the inhibitors and the corresponding association constants (k(ass)) provide additional validation for the structures. Moreover, some of the biochemical differences observed between the vivapains may be explained by the results of the docking studies. The overall structures of the two vivapains are similar to each other as well as to the falcipains with most of the catalytic residues conserved. At the same time, some important differences are observed between the sizes of the binding pockets as well as some of the residues involved in binding. The study suggests a likelihood of developing common inhibitors for these enzymes provided the interesting differences in the binding pockets of these enzymes are critically considered during such an attempt. The results of the current study can be utilized in de novo drug design for effective treatment of malaria.

Influence of proline on rat brain activities of alanine aminotransferase, aspartate aminotransferase and acid phosphatase.

Hyperprolinemia type II (HPII) is an autosomal recessive disorder caused by the severe deficiency of enzyme delta1-pyrroline-5-carboxylic acid dehydrogenase leading to tissue accumulation of proline. Chronic administration of Pro led to significant reduction of cytosolic ALT activity of olfactory lobes (50.57%), cerebrum (40%) and medulla oblongata (13.71%) only. Whereas mitochondrial ALT activity was reduced significantly in, all brain regions such as olfactory lobes (73.23%), cerebrum (70.26%), cerebellum (65.39%) and medulla oblongata (65.18%). The effect of chronic Pro administration on cytosolic AST activity was also determined. The cytosolic AST activity from olfactory lobes, cerebrum and medulla oblongata reduced by 75.71, 67.53 and 76.13%, respectively while cytosolic AST activity from cerebellum increased by 28.05%. The mitochondrial AST activity lowered in olfactory lobes (by 72.45%), cerebrum (by 78%), cerebellum (by 49.56%) and medulla oblongata (by 69.30%). In vitro studies also showed increase in brain tissue proline and decrease in glutamate levels. In vitro studies indicated that proline has direct inhibitory effect on these enzymes and glutamate levels in brain tissue showed positive correlation with AST and ALT activities. Acid phosphatase (ACP) activity reduced significantly in olfactory lobes (40.33%) and cerebrum (20.82%) whereas it elevated in cerebellum (97.32%) and medulla oblongata (76.33%). The histological studies showed degenerative changes in brain. Following proline treatment, the animals became sluggish and showed low responses to tail pricks and lifting by tails and showed impaired balancing. These observations indicate influence of proline on AST, ALT and ACP activities of different brain regions leading to lesser synthesis of glutamate thereby causing neurological dysfunctions.

Usefulness of the admission electrocardiogram for identifying the infarct-related artery in inferior wall acute myocardial infarction.

We analyzed the admission electrocardiogram in 109 patients with inferior wall acute myocardial infarction in an attempt to determine the infarct-related artery (IRA). The presence of ST depression in leads V1 or V2 had a high sensitivity for predicting the left circumflex artery as the IRA. The lack of ST depression in V1 or V2 had a high negative predictive value for excluding the left circumflex artery as the IRA.

Can audit improve the quality of care for routine tonsillectomy?

Tonsillectomy is the most commonly performed operation in otolaryngology practice. In order to assess outcome and improve quality of care, 65 patients were asked to answer a post-operative questionnaire. The questionnaire was then audited and patient management changed. Following this, another group of 65 patients revealed, in the same questionnaire, a significant improvement in care.

Compensation for refractive-index variations in laser Doppler anemometry.

Techniques to compensate for index of refraction variations in the application of a laser Doppler anemometer are examined. For discontinuous plane-layered media a method of discrete elements is employed. An alternative set of equations is derived for continuous cylindrically layered media and simplified to make comparisons with available results for a single cylinder. Results of velocity measurements in a Plexiglas model of a cylindrically layered nuclear fuel assembly are presented to establish a positioning accuracy of the method to within 0.025 cm.