Types of Rotavirus Causing Acute Diarrhea among Children in Western India, their Demographic Pattern and Disease Severity.

Background: Rotavirus infection is a major cause of severe acute gastroenteritis among infants and children all over the world1 with winter out-breaks of diarrhea in temperate and cooler parts almost round the year. However, this varies in different part of India.2-6 Diarrhea is a major cause of under-5 mortality, contributing to approximately over 1,50,000 infant deaths in our country per year.15,16 Different genotypes have been identified and many more are emerging by way of mutation, genetic shift and genetic drifts. Rotavirus are classified antigenically as A (Most common), B, C, D, E by ELISA and genotypically as G (1 through 12) and P (1 through 8) by Reverse Trans criptase PCR, in combinations. Materials and methods: Stool samples of 110 infants and children from 6 to 60 months of age, with suspected viral diarrhea over one year period were studied for serotypes and genotypes; and compared for their respective disease severity. Results: Thirty-four percent were found positive for Rotavirus-A by ELISA. Of the positive, 33.4% were found to be of G9 genotype, much higher than reported from other parts of the country. On the other hand, merely 13.6% of G1 and G4 each were detected, contrary to high prevalence elsewhere. On electro-pherotyping, the long-arm types were associated with more severe disease (64.6% showing moderate to severe dehydration) than their short-arm types (Only 16.6% showed moderate dehydration only) p < 0.009. No difference in incidence of severe dehydration between AD positive for Rotavirus (11.7%) and those found nega tive (11.8%), presumably due to other viruses, after excluding invasive diarrhea. Conclusion: Emergence of diverse strains, i.e. more of G9 and G12 genotypes than earlier reports of G1 and G2 types indicate considerable genetic shift in the region. Such trend could have significant implication on degree of seroconversion from currently used live vaccines, using G1 or bovine reassortant G1-3 strains only, seen in recent studies from Africa and Malayi.29 Contrary to claims that Rotavirus diarrhea usually threatened severe diarrhea, no significant difference in incidence of severe diarrhea was observed between Rotavirus positive and Rotavirus negative acute diarrhea.

Factors affecting induction and development of in vitro rooting in apple rootstocks.

Shoots of apple rootstocks raised in vitro were transferred to various rooting media to study the effect of different factors on root initiation and development. Various concentrations of indole-3-butyric acid (IBA) initiated rooting but maximum rooting percentage was found with 2.0 and 2.5 mg l(-1) of IBA in M7 and with 1.0 mg l(-1) of IBA in MM106. The drawback was that the roots were thick, short and with profuse callus. The presence of activated charcoal (AC) in the rooting medium improved the rooting quality but reduced the rooting percentage in both the rootstocks. In high auxin dip of 70, 80 and 90 mg l(-1) IBA for 2, 2 and 1 hr showed 75-85 per cent rooting in M7, but lacked reproducibility of the results. Whereas in MM106, 66 - 70 % rooting was achieved with 70 mg l(-1) of IBA dip for 3 h. Root induction in shoots in IBA containing liquid medium (LM) in dark for few days and root elongation in IBA--free medium in light proved most effective. On the other hand, continuous light treatment showed reduced rooting. Reduction of MS salts and sucrose in root elongation medium showed decreased rooting. Plantlets from two--stage rooting procedure showed more rapid growth and satisfactory survival during hardening of plants and on transfer to field.

Pharmacovigilance in kala-azar patients with severe thrombocytopenia caused by sodium antimony gluconate & miltefosine.

Sodium antimony gluconate (SAG) and miltefosine used in the treatment of kala-azar are known to cause several side effects but severe thrombocytopenia has not been reported. Four cases of severe thrombocytopenia, two caused by SAG and two by miltefosine were promptly detected and treated by immediate withdrawal of the offending drugs, platelet and blood transfusions and dexamethasone. After improvement Leishman-Donovan (LD) bodies were demonstrated in splenic aspirates of both patients of SAG group and one of miltefosine and they were treated with 1 mg/kg body wt of amphotericin B for 20 days and cured. One patient of miltefosine group treated outside only on the basis of rK-39 positivity did not show LD bodies in splenic aspirates and improved without any antikala- azar drug. None of the patients relapsed within 6 months of follow up. Prompt detection of side effects under the concept of pharmacovigilance can save life of such patients.

In vitro clonal multiplication of an apple rootstock by culture of shoot apices and axillary buds.

In vitro clonal multiplication of apple rootstock MM 111 using axillary buds and shoot apices were carried out. Vegetative axillary buds of the size of 0.2-2.0 cm and shoot apices measuring 4 mm in length were initiated to shoot proliferation on MS medium supplemented with BA (0.5 - 1.0 mgl(-1)), GA3(0.5 mgl(-1)), with or without IBA(0.05 - 0.1 mgl(-1)). Small size explants showed less phenol exudation and less contamination. Following establishment phase, the small shoots emerged from explants were subcultured on MS medium supplemented with different combinations and concentrations of growth regulators. BA (1.0 mgl(-1)) and GA3 (0.5 mgl(-1)) combination showed highest multiplication rate (1:5), andcl also produced longer shoots. Two step rooting was done by transferring microcuttings to auxin free solid medium after root initiation in dark on 1/2 strength MS liquid medium containing IBA (0.5 mgl(-1) ). Rooted plantlets were transferred to peat containing paper cups and resulting plants of MM 111 acclimated successfully for transfer to field.

Immobilization of Amaranthus leaf oxalate oxidase on alkylamine glass.

A membrane bound oxalate oxidase from leaves of Amaranthus spionsus has been partially purified and immobilized on alkylamine glass with a yield of 9.2 mg protein/g support. The enzyme retained 99.4% of initial activity of free enzyme after immobilization. There was no change in the optimum pH (3.5) and Vmax but the temperature for maximum activity was slightly decreased (35 degrees C) and energy of activation (Ea) and Km for oxalate were increased after immobilization. The immobilized enzyme preparation was stable for 6 months, when stored in distilled water at 4 degrees C. Presence of Cl- did not affect the activity of immobilized enzyme.

Determination of plasma oxalate with chloride ion insensitive oxalate oxidase.

A simple colorimetric method has been developed for determination of oxalate in plasma using a C1-insensitive oxalate oxidase purified from grain sorghum leaves. The ultrafiltered plasma collected in 3.5 N HC1 was pretreated with NaNO2 to avoid possible interference by ascorbate. The minimum detection limit of the method is 0.225 mg/l. The % recovery of the added oxalate was 91.5 +/- 5.0 (mean +/- S.D., n = 15). The coefficient of variation within and between batch were < 3 and < 5 respectively. The mean plasma oxalate concentration in healthy subjects was 0.29 mg/l. The method has the advantage over other enzymic methods that it doesn't require the removal of C1- prior to oxalate assay.

Chemical analysis of urinary calculi in Haryana.

Quantitative chemical analysis of 225 urinary calculi (128 Renal, 57 bladder and 40 ureter) collected from hospitals of different districts of Haryana was carried out. CaOxM was found to be present as a major component in all the stones analysed, while MAP, HA, CA and UA were found in 83.7%, 83.6%, 13% and 78.9% cases, respectively. The content of CaOxM was found to be higher in renal stones as compared to ureter and bladder stones, while the concentration of MAP was higher in bladder stones as compared to kidney and ureter stones. HA content was higher in ureter and UA was higher in bladder stones as compared to other urinary stones.

Adenosine triphosphate content of Mycobacterium leprae by percoll buoyant density centrifugation.

Thirty nine untreated patients of bacilliferous leprosy with a mean bacteriological index of 4.8 and morphological index of 1.3% formed the study group. Adenosine triphosphate assay was carried out by (i) enzyme treatment method in 18 patients and (ii) percoll buoyant density gradient method in 21 patients. ATP content obtained by percoll buoyant density gradient method was significantly higher than that obtained by enzyme treatment method. Percoll buoyant density centrifugation for purification and isolation of bacilli from human leproma is simplier, quicker and can serve as an alternate method of enzyme treatment.

Primary dapsone resistance as assessed by uptake of labelled thymidine by the macrophage resident Mycobacterium leprae.

12 untreated lepromatous leprosy patients were screened for primary dapsone resistance by the uptake of labelled thymidine by macrophage resident M. leprae. There were found to harbour primary dapsone resistant strains of M. leprae and another three partially resistant strains to the drug. This rapid, simple and reliable method should be used routinely to screen leprosy patients, for drug resistance.

Effect of suppressor cells on antibody producing cells in mice infected with Mycobacterium leprae.

Swiss albino mice were transfused with suppressor cells obtained after in vivo stimulation of mice with Con A (NS group). Some of the animals were infected with Mycobacterium leprae (NSI-group). Half of these animals were treated with dapsone (NSIT group). Adequate normal (NC) and infected (NI) controls were included. A plaque assay was carried out at different time periods to elucidate the effect of suppressor cells on antibody producing cells. No significant difference was seen in the number of plaque forming cells (PFC) in infected and dapsone treated animals (NSIT) when these were compared with controls. However significant increase seen in the number of IgM plaque forming cells at 6 months in NI and NSI groups and IgG PFC in NI group could be due to the peak footpad infection during this period. The significant decrease in the number of IgG PFC in NS and NSIT group compared to NC at 0 month is probably due to the suppressor cell activity in these groups.

Analysis of nucleosome arrangement on satellite DNA of rat liver chromatin.

The arrangement of nucleosomes on the nucleotide sequence of satellite DNA of Oceanian rat (Rattus rattus) has been studied. Nucleosome cores were prepared from rat liver nuclei with micrococcal nuclease, exonuclease III and nuclease Sl. From the total population of core DNA fragments, the satellite-containing fragments were selected by molecular cloning and the complete nucleotide sequence of these clones was determined. The data show that nucleosomes occupy a number of preferred positions on satellite DNA. These positions are strictly defined. Thus location of nucleosomes along the satellite sequence is non-random. Such finding may have important biological significance.

Levels of the third component of complement in Mycobacterium leprae infected intact and immune compromised mice.

Normal and immunosuppressed mice were infected with Mycobacterium leprae and the bacillary counts were made from the footpads at 3, 6 and 9 months post inoculation. A decrease in the serum C3 level was observed in the infected groups of animals compared to controls.