Induction of giant cells by the synthetic food colorants viz. lemon yellow and orange red

Cytotoxicity and giant cell formation induced by lemon yellow and orange red synthetic food colorants were evaluated in the present study. The aqueous solutions of both the dye solutions were tested for cytotoxicity using Allium cepa assay. Frequency of giant cells were determined after treating the root tips with different concentrations of both food colorant solutions viz., 0.005, 0.01, 0.05, 0.1 % for varying time durations (1/2, 1, 2, 3 h). These colorants may cause giant cell formation primarily by interfering with the normal course of mitosis. Giant cells showing multiple aberrations viz. bridged and binucleate condition, cellular fragmentation, nuclear lesion, double and multiple nuclear lesions, double nuclear peaks and cellular breakage, elongated nucleus, nuclear budding, hyperchromasia, micronucleus, nuclear erosion, pulverized nucleus etc. were induced in root tips treated with both of the colorants. The synthetic food colorant treated cells showed inhibition of cell division and induction of giant cells. A dose dependant decrease in the mitotic index [88.20 % (c^?ve, 3h) to 81.54 % (Lx₄, 3h) and 88.20 % (c^?ve, 3h) to 73.17 % (Ox₄, 3h)] was observed. All mitotic phases show significant induction of giant cells when treated with both food colorants. Interphase stage shows higher percentage of giant cells, whereas in cytokinesis it was negligible. The orange red food colorant is observed to be more toxic because it recorded higher percentage of giant cell induction when compared with lemon yellow [27.93 % (Lx₄, 3h) and 28.07 % (Ox₄, 3h)].     

Histoenzymological demonstration of alkaline and acid phosphatases in the intestinal mucosa of several birds, viz Ploceus philippinus, Megalaima haemacephala and Halcyon smyrnensis.

Alkaline and acid phosphatases were histochemically localized in the ileum of three birds with diverse feeding habits. The birds chosen for study included the granivorous Ploceus philippinus, the frugivorous Megalaima haemacephala, and the mainly piscivorous Halcyon smyrnensis. It was found the the activity of these phosphatases tend to be greater in H. smyrnensis and M. haemacephala and comparatively less in P. philippinus, as demoted by variable enzymatic deposition. Simultaneously, their functional significance has been discussed, correlating enzymatic activities with the specific feeding habits of the birds.     

Observations on a new species of Symphyonemopsis Tiwari & Mitra,viz. S. Pantii n. sp. (Cyanophyta,Stigonematales)

The paper describes the morphology and reproduction of a new species of Symphyonemopsis viz. S. pantii n. sp. isolated from enrichment cultures of a soil sample collected from the campus of Sangeet Samiti, Allahabad (India). Thalli grow as dark blue-green, small, wooly patches on soil surface but as cushion-like growths on agar plates. It has a prostrate and erect system. The latter bears secondary branches of almost equal breadth. It possesses both false and true branches. Usually true branches are reverse V-shaped. The alga shows formation of hormocysts and akinetes. In old cultures usually the main filament becomes multiseriate. The crescent-shaped stages of hormocysts are also present here. Cells of the present alga are constricted at the cross-walls On comparison it was found to come close to Symphyonemopsis katniensis Tiwari et Mitra but differs in certain fundamental characters. It is, therefore, described as a new species viz. S. pantii n. sp. (Cyanophyta, Stigonematales, Mastigocladaceae).     

Isolation and purification of an extracellular protease from a new strain of Bacillus subtilis, viz.NCIM 2711

A new extracellular protease having a prospective application in the food industry was isolated from Bacillus sUbtilis NCIM 2711 by (NH4)2SO4 precipitation from the cell broth. It was purified using DEAE-Cellulose and CM-Sephadex C-50 ion-exchange chromatography. With casein as a substrate, the proteolytic activity of the purified protease was found to be optimal at pH 7.0 and temperature 55 degrees C with Km 1.06 mg/ml. The enzyme was stable over a pH range 6.5-8.0 at 30 degrees C for 1 hr in presence of CaCl2 x 2H2O. At 55 degrees C, the enzyme retained 60% activity up to 15 min in presence of CaCl2 x 2H2O. EDTA and o-phenanthroline (OP) completely inhibited the enzyme activity while DFP, PMSF and iodoacetamide were ineffective. The enzyme was completely inhibited by Hg2+ and partially by Cd2+, Cu2+, Ni2+, Pb2+ and Fe2+. The OP inhibited enzyme could be reactivated by Zn2+ and Co2+ up to 75% and 69% respectively. It is a neutral metalloprotease showing a single band of 43 kDa on SDS-PAGE.     

Changes in rodent-erythrocyte methemoglobin reductase system produced by two malaria parasites, viz. Plasmodium yoelii nigeriensis and Plasmodium berghei

The methemoglobin reductase system plays a vital role in maintaining the equilibrium between hemoglobin and methemoglobin in blood. Exposure of red blood cells to oxidative stress (pathological/physiological) may cause impairment to this equilibrium. We studied the status of erythrocytic methemoglobin and the related reductase system during Plasmodium yoelii nigeriensis infection in mice and P. berghei infection in mastomys. Malaria infection was induced by intraperitoneal inoculation with 10⁶ infected erythrocytes. The present investigation revealed a significant decrease in the activity of methemoglobin reductase, with a concomitant rise in methemoglobin content during P. yoelii nigeriensis infection in mice erythrocytes. This was accompanied with a significant increase in reduced glutathione and ascorbate levels. The activity of lactate dehydrogenase, glucose 6-phosphate dehydrogenase and glutathione reductase increased with a progressive rise in parasitemia. However, no methemoglobin or associated reductase activity was detected in normal and P. berghei-infected mastomys. P. berghei infection in mastomys resulted in an increase in the level of reduced glutathione and ascorbate in erythrocytes, and also in the activity of lactate dehydrogenase, glucose 6-phosphate dehydrogenase and glutathione reductase. These results suggest that antioxidants/antioxidant enzymes may prevent or reduce the formation of methemoglobin in the host and thereby protect the host from methemoglobinemia.     

Infection of Bacterial Endosymbionts in Insects: A Comparative Study of Two Techniques viz PCR and FISH for Detection and Localization of Symbionts in Whitefly,Bemisia tabaci

Bacterial endosymbionts have been associated with arthropods and large number of the insect species show interaction with such bacteria. Different approaches have been used to understand such symbiont- host interactions. The whitefly, Bemisia tabaci, a highly invasive agricultural pest, harbors as many as seven different bacterial endosymbionts. These bacterial endosymbionts are known to provide various nutritional, physiological, environmental and evolutionary benefits to its insect host. In this study, we have tried to compare two techniques, Polymerase chain reaction (PCR) and Flourescence in situ Hybridisation (FISH) commonly used for identification and localization of bacterial endosymbionts in B. tabaci as it harbors one of the highest numbers of endosymbionts which have helped it in becoming a successful global invasive agricultural pest. The amplified PCR products were observed as bands on agarose gel by electrophoresis while the FISH samples were mounted on slides and observed under confocal microscope. Analysis of results obtained by these two techniques revealed the advantages of FISH over PCR. On a short note, performing FISH, using LNA probes proved to be more sensitive and informative for identification as well as localization of bacterial endosymbionts in B. tabaci than relying on PCR. This study would help in designing more efficient experiments based on much reliable detection procedure and studying the role of endosymbionts in insects.     

In vitro synthesis of a specific protein viz. Asparagine synthetase by nuclear fraction from gamma irradiated potato buds

Irradiation induced synthesis of asparagine synthetase was localized in the nucleus of the potato buds. Isolated nuclei were capable of incorporating amino acids into asparagine synthetase protein and the process was inhibited by cycloheximide (100%), puromycin (73%) and by feed back inhibitor β-aspartyl hydroxamate (100%). Preincubation of the bud tissue with actinomycin D was necessary to express its inhibitory effect.     

Characterization of the 3-HKT gene in important malaria vectors in India, viz: Anopheles culicifacies and Anopheles stephensi (Diptera: Culicidae)

The 3-hydroxykynurenine transaminase (3-HKT) gene plays a vital role in the development of malaria parasites by participating in the synthesis of xanthurenic acid, which is involved in the exflagellation of microgametocytes in the midgut of malaria vector species. The 3-HKT enzyme is involved in the tryptophan metabolism of Anophelines. The gene had been studied in the important global malaria vector, Anopheles gambiae. In this report, we have conducted a preliminary investigation to characterize this gene in the two important vector species of malaria in India, Anopheles culicifacies and Anopheles stephensi. The analysis of the genetic structure of this gene in these species revealed high homology with the An. gambiae gene. However, four non-synonymous mutations in An. stephensi and seven in An. culicifacies sequences were noted in the exons 1 and 2 of the gene; the implication of these mutations on enzyme structure remains to be explored.