Assessment of Sugarcane Germplasm ( Saccharum spp. complex) Against Red Rot Pathogen Colletotrichum Falcatum

ABSTRACT Red rot, caused by Colletotrichum falcatum Went is the most important disease of sugarcane in India inflicting substantial loss to both cane industry and cane growers. To keep in view the importance of red rot disease of sugarcane, 117 accession of sugarcane germplasm including different Saccharum species and Indian and foreign commercial hybrids were tested against red rot with Cf 07, Cf 08 & Cf 09 (national pathotypes) by plug method of inoculation. Out of 117, 6 were found resistant and 12 were moderately resistant against red rot and rest were moderately susceptible/susceptible/highly susceptible. Theses resistance and moderately resistant accession can be further utilize to produce resistance varieties against the most devastating pathogen of sugarcane.

Characterization and evaluation of apoptotic potential of double gene construct pVIVO.VP3.NS1.

Viral gene oncotherapy, targeted killing of cancer cells by viral genes, is an emerging non-infectious therapeutic cancer treatment modality. Chemo and radiotherapy in cancer treatment is limited due to their genotoxic side effects on healthy cells and need of functional p53, which is mutated in most of the cancers. VP3 (apoptin) of chicken infectious anaemia (CIA) and NS1 (Non structural protein 1) of Canine Parvovirus-2 (CPV-2) have been proven to have oncolytic potential in our laboratory. To evaluate oncolytic potential of VP3 and NS1 together these genes needed to be cloned in a bicistronic vector. In this study, both these genes were cloned and characterized for expression of their gene products and its apoptotic potential. The expression of VP3 and NS1 was studied by confocal microscopy and flowcytometry. Expression of VP3 and NS1 in pVIVO.VP3.NS1 transfected HeLa cells in comparison to mock transfected cells indicated that the double gene construct expresses both the products. This was further confirmed by flowcytometry where there was increase in cells expressing VP3 and NS1 in pVIVO.VP3.NS1 transfected group in comparison with the mock control group. The apoptotic inducing potential of this characterized pVIVO.VP3.NS1 was evaluated in human cervical cancer cell line (HeLa) by DNA fragmentation assay, TUNEL assay and Hoechst staning. This double construct was observed to induce apoptosis in HeLa cells.

Synthesis of multiple biologically active 1, 2-dihydro-pyrimido [1, 2-A]-benzimidazole-3- carbonitrile.

A variety of 1,2-dihydropyrimido-[1,2-a]-benzimidazole-3-carbonitrile derivatives were synthesized under microwave irradiation using water and acetonitril as solvent system. All the compounds were tested in vitro for α-glucosidase inhibitory and DPPH free radical scavenging activity. 4-Amino-2-(4-flourophenyl)-1,2-dihydropyrimido [1,2-a]-benzimidazole-3-carbonitrile (4c) was found to be a potent intestinal α- glucosidase inhibi tory activity (IC50; 91μM) along with moderate DPPH scavenging property. This compound was further evaluated for cytotoxicity activity against HT-29 colon cancer cell line. The IC50 value for its cytotoxicity activity was found to be 662 μM.

A report on the indoor residual spraying (IRS) in the control of Phlebotomus argentipes, the vector of visceral leishmaniasis in Bihar (India): an initiative towards total elimination targeting 2015 (Series-1).

Background & objectives: Visceral leishmaniasis, commonly known as kala-azar is endemic in Bihar state, India. Current vector control programme in Bihar focuses mainly on spraying the sandfly infested dwellings with DDT. The Government of India in collaboration with WHO has fixed the target 2015 for total elimination of kala-azar. The present study was carried out to see the impact of DDT and improved IEC in the containment of vector density vis-à-vis disease transmission. Methods: Before the start of the spraying operations training was imparted to all the medical and paramedical personnel regarding the methods of spraying operations. Pre- and post-sandfly density was monitored in four selected districts. Incidences of kala-azar cases were compared for pre- and post-spray periods. Social acceptability and perceptions of households was collected through questionnaires from 500 randomly selected households in the study districts. Results: House index in three study districts reduced considerably during post-spray when compared to pre-spray. Kala-azar incidence in many districts was reduced after the DDT spray. Either partial or complete refusal was reported in 14.4%, while 35% were not satisfied with the suspension concentration and coverage; and 46.6% were found satisfied with the spraying procedure. Interpretation & conclusion: Strengthening the IEC activities to sensitise the community, proper training of health personnel, monitoring of spray, good surveillance, proper treatment of cases and two rounds of DDT spray with good coverage in the endemic districts up to three years are essential to achieve the desired total elimination of kala-azar in Bihar state.

Opinion: Pathophysiological approach to the management of scorpion envenomation.

Indian red scorpion (Mesobuthus tamulus; MBT) produces lethal stings and is a matter of concern in certain parts of India. MBT envenomation produces multi-systemic involvement, thus presents difficulty in the management. Symptomatic treatment has been practiced earlier that failed to relieve the toxic effects of the venom. Therefore, present manuscript deals with pathophysiologically based approach in the management of toxicity considering the merits and the demerits of treatment protocols so as to evolve a consensus in the treatment strategies of scorpion envenomation.

Characterisation of fowl adenoviruses from chickens affected with infectious hydropericardium during 1994-1998 in India.

In the present study characterisation has been done for six group I fowl adenoviruses (FAV) isolated from outbreaks of infectious hydropericardium (IHP) of chickens that occurred in different states/regions of India during the years 1994-98. These six viruses were identified as FAV serotype 4 by virus neutralisation and restriction endonuclease analyses. Antigenic analyses of the viruses revealed close relationship (R-values 0.93-0.96). Under the experimental conditions, we have been able to induce IHP using FAV serotype 4 isolate AD: 411 and were also able detect FAV antigens in myocardial tissues by immunofluorescence assay (a new observation), an indication that IHP causing FAV serotype 4 strain replicate in myocardial tissue. Restriction endonuclease analysis of the viral genomes (approximately 46 Kb), using Hind III, Sma I, Xba I, Bam HI, Pst I and Dra I produced identical genetic profiles. Pst I and Bam HI profiles for these six vitus isolates were identical to those published earlier for an IHP causing Pakistani FAV serotype 4 isolate KR31. The identical genetic profiles of viruses, chronology of the outbreaks of IHP in Pakistan during 1989 onward and later in Jammu and Kashmir, India (1994), suggest that FAV serotype 4 isolates involved in outbreaks of IHP in India had probably spread from Pakistan. In order to prevent further spread and economic losses due to IHP in India, based on the antigenic relatedness data in this paper, any one of the six studied FAV serotype 4 isolates can be used as a candidate for mass production of CEH culture based killed vaccine.

Purification and differentiation of three avian adenoviruses by restriction endonuclease analysis.

A rapid method of ultracentrifugation pelleting of avian adenovirus (AAV) from small volume of chloroform treated infected cell culture fluid or allantoic fluid was adapted for isolation of adenoviral DNA. The viral DNA extracted from semipurified viruses was found to be intact on agarose gel and pure enough (A260/280 = 1.85-1.92) for restriction enzyme analysis. Restriction endonuclease analysis of Indian strain of AAV serotype 1, AAV serotype 4 (group I AAVs) and egg drop syndrome-76 (EDS-76) virus genomes (group III AAV) with Hind III enzyme differentiated these viruses. The AAV serotype 1 and serotype 4 strain exhibited identical Hind III profile to European viral strains belonging to same serotypes however, the EDS-76 virus gave similar but not identical profile. The calculated genomic lengths for AAV serotype 1 and EDS-76 virus were approximately found to be 33.9 and 44.4 Kb, respectively.

Detection of bluetongue virus genome segment 6 sequences by RT-PCR.

Reverse transcription-PCR (RT-PCR) technique was adopted to amplify a 101 basepair nucleotide sequence of bluetongue virus (BTV) genome segment 6. The specificity of the amplicon was determined by its approximate size in 3% agarose gel electrophoresis, digestion with restriction enzyme MspI, dot-blot hybridization and cycle sequencing. The technique was found to be suitable for detection of bluetongue virus in infected cell culture and clinical samples.

Toxigenicity of Edwardsiella tarda isolates of fish and pig origin in experimental models.

Indian isolates of E. tarda from fish (7) and pigs (2) were examined for their enterotoxigenicity/cytotoxigenicity in rabbit ligated ileal loop (RLIL), suckling mouse, rabbit skin and Vero cell monolayers. Cell free culture filtrates (CFCF) of isolates from fish and pigs, induced blood tinged fluid accumulation in RLIL, increased vasopermeability in rabbit skin, and caused cytopathic effect in Vero cells but could not induce fluid accumulation in suckling mouse. CFCFs lost their activity on heating at 63 degrees C for 30 min or 72 degrees C for 15 sec, and also at pH < or = 4.5 or > or = 8.5. The toxic factor was released extracellularly and was nondializable.

Augmentation of phenyldiguanide-induced bradycardia by Buthus tamulus venom in adult rats.

Intravenous injection of phenyldiguanide (PDG) in anaesthetized rats produced dose-dependent (1-10 micrograms/kg) decrease in heart rate for a period of time (time-response area). The maximum response occurred at 10 micrograms/kg PDG. Administration of B. tamulus (BT) venom as low as 20 micrograms/kg augmented the PDG-induced bradycardia response by 2.5 times the initial PDG (10 micrograms/kg) response. The maximal augmentation was observed after 60 min of venom injection. Increasing the BT venom concentration to 40 micrograms/kg failed to enhance the reflex response (1.7 times the initial response). The threshold concentration of BT venom was 4 micrograms/kg. BT venom (100 micrograms/kg) alone, decreased the heart rate significantly only after 90 min. Results indicate that, even the sublethal concentrations of BT venom sensitize the reflexes elicited by PDG.

Effect of cigarette smoking on serum total cholesterol and high density lipoprotein cholesterol in normal subjects and coronary heart patients.

In the present study, 51 apparently normal volunteers and 34 clinically established coronary heart disease cases were studied. 21 out of 51 normal, and 16 out 34 coronary heart disease cases were cigarette smokers. The cases were divided in two age groups--20 to 40 years, the younger age group and 41 to 60 years, the older age group. Total serum cholesterol and high-density lipoprotein cholesterol of all the cases were determined. The ratio of total cholesterol to high-density lipoprotein cholesterol was significantly higher in all the smoking normal and coronary heart disease smokers. Hence, the higher levels of total cholesterol to high density lipoprotein cholesterol ratio appears to be one of the important parameters to ascertain the development of coronary heart disease in cigarette smokers.