PubMed Health⌕ Search

Biomedical subjects

G Chandra

Publications and source records attributed to G Chandra.

At least 55 records · Page 3Linked to original sources

Studies on the filariasis vector--Culex quinquefasciatus at Kanchrapara, west Bengal (India).

A nine month entomological and parasitological study was conducted between December 1991 and August 1992, at Kanchrapara, a town of the district North 24-Parganas, West Bengal. The town was found to be endemic for bancroftian filariasis. Microfilaria rate among the human population was 1.9 per cent (35 of 1200 persons). Average man hour density of C. quinquefasciatus was considerably high (31.10) and a higher vector prevalence was seen in summer. The infection and infectivity rates of the natural population of the vector were 2.3 and 0.28 per cent respectively. 9.2 and 1.7 per cent of the human habitations were found to contain infected and infective C. quinquefasciatus. Both infection and infectivity rates were higher during the rainy season than the other seasons.

Adult↗

Correlation between the preferred biting site of Culex quinquefasciatus and the region of the body affected by clinical filariasis.

Entomological and filariasis surveys were conducted simultaneously in a rural and an urban environment in West Bengal from September 1986 to August 1987. The aim of the surveys was to see if any correlation existed between the preferred biting site of Culex quinquefasciatus, the filarial vector, and the region of the body affected by clinical filariasis. Significantly more mosquitoes (> 55%) attacked the thighs, legs and feet of human volunteers than other sites on their bodies, whether the volunteers were indoors or outdoors, in rural or urban environments. Significantly more patients had clinical filariasis of these lower body parts than of the upper parts. The vector's preferred biting sites and the anatomical sites most affected by clinical filariasis were therefore the same.

Adult↗

A CRE/ATF-like site in the upstream regulatory sequence of the human interleukin 1 beta gene is necessary for induction in U937 and THP-1 monocytic cell lines.

Transfection of U937 and THP-1 cells with a recombinant plasmid, pIL1(4.0kb)-CAT, containing 4 kb of the interleukin 1 beta (IL-1 beta) gene upstream regulatory sequence resulted in inducer-dependent expression of chloramphenicol acetyltransferase activity. Treatment of the transfected cells with various combinations of the inducers lipopolysaccharide, phorbol myristate acetate, and dibutyryl cyclic AMP upregulated the IL-1 beta promoter. In U937 and THP-1 cells, maximum stimulation of both the endogenous IL-1 beta gene and pIL1(4.0kb)-CAT transfectants was observed following treatment with the combination of inducing agents lipopolysaccharide-phorbol myristate acetate-dibutyryl cyclic AMP. This combination of inducing agents was used to identify and study, at the molecular level, some of the regulatory elements necessary for induction of the IL-1 beta gene. A series of 5' deletion derivatives of the upstream regulatory sequence were used in transient transfection assays to identify an 80-bp fragment located between -2720 and -2800 bp upstream of the mRNA start site that was required for induction. Exonuclease III mapping, electrophoretic mobility shift assays (EMSA), and DNA sequence analysis of this region were used to identify a transcription factor binding sequence which contained a potential cyclic AMP response element (CRE/ATF)- and NF-kappa B-like binding site. Site-directed mutagenesis of the CRE/ATF-like site resulted in the loss of binding of a specific factor or factors as determined by EMSA. The loss of binding activity directly correlated with a loss of approximately 75% of promoter activity as determined in transient transfection assays. As determined by EMSA, the factor binding to the CRE/ATF-like site was present in nuclear extracts prepared from both uninduced and induced THP-1 and U937 cells. However, the intensity of the band appeared to be increased when nuclear extracts from induced cells were used. In contrast to the CRE/ATF mutation, which resulted in the loss of promoter activity, mutation of the NF-kappa B-like site resulted in a moderate increase in activity in U937 cells. A similar increase in promoter activity was not observed in THP-1 cells. From these studies, we conclude that a CRE/ATF-like site and a factor or factors interacting with this site are essential for the maximum induction of the IL-1 beta gene in stimulated U937 and THP-1 cells.

Activating Transcription Factors↗

Large-scale purification of plasmid DNA by fast protein liquid chromatography using a Hi-Load Q Sepharose column.

The large-scale purification of plasmid DNA was achieved using fast protein liquid chromatography on a Hi-Load Q Sepharose column. This method allows for the purification of plasmids starting from crude plasmid DNA, prepared by a simple alkaline lysis procedure, to pure DNA in less than 5 h. In contrast to the previously described plasmid purification methods of CsCl gradient centrifugation or high-pressure liquid chromatography, this method does not require the use of any hazardous or expensive chemicals. More than 100 plasmids varying in size from 3 to 15 kb have been purified using this procedure. A Mono Q Sepharose column was initially used to purify plasmids smaller than 8.0 kb; however, a Hi-Load Q Sepharose column proved more effective with plasmids larger than 8 kb. The loading of plasmids larger than 8 kb on the Mono Q column resulted in a high back pressure and the plasmid DNA could not be eluted from the column. Thus, for routine purification we utilize the Hi-Load Q Sepharose column. Plasmids purified by this method had purity, yield, and transfection efficiency in mammalian cells similar to those of plasmids purified by CsCl density gradient centrifugation.

Centrifugation, Density Gradient↗