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Biomedical subjects

M Verma

Publications and source records attributed to M Verma.

At least 55 records · Page 3Linked to original sources
PubMed1996

Promoter of the canine tracheobronchial mucin gene.

The mucin gene is up-regulated in diseases such as cystic fibrosis (CF) and asthma. To understand the mechanisms involved in transcriptional regulation of mucin gene expression we have characterized the region of the mucin gene up-stream of the transcriptional start site and analysed the cis-acting elements required for mucin promoter activity. We isolated clones from a dog genomic library containing the promoter region for the tracheobronchial mucin gene (TBM). The authenticity of the promoter was tested by nucleotide sequencing, primer extension analysis, electrophoretic mobility shift assay (EMSA) and reporter gene expression analysis. The canine TBM promoter is different from housekeeping gene promoters (as it is not rich in GC content and contains TATA- and CAAT-like sequences) and different from that of regulatory genes (because it contains many TATA- and CAAT-like sequences and multiple transcriptional initiation sites). Reporter gene analysis using canine TBM promoter-chloramphenicol acetyltransferase (CAT) fusion plasmids established the regions responsible for promoter activity and verified the positions of the major mucin transcriptional initiation sites. Reporter gene analysis also established that a region of the canine TBM promoter and first exon containing all of the transcriptional initiation sites is more active in mucin expressing cells (e.g. CT1 cells-immortalized canine tracheal epithelial cells, human CFT1 cells-immortalized tracheal epithelial cells from a CF subject, or HBE1 cells-immortalized tracheal epithelial cells from non-CF subject) than in mucin non-expressing cells (COS7, 3T3), suggesting cell specificity. The promoter region contained cAMP response element (CRE) sequences, and the TBM gene transcription was enhanced when cAMP analogs were added to transfected cells. EMSA indicated the presence of at least two DNA binding proteins in CT1 cells. This is the first report describing the characterization of a TBM gene promoter. The information obtained in the present studies will be valuable in understanding mucin gene regulation in normal and pathological conditions.

Animals↗
PubMed1996

Tracheo-bronchial mucin gene expression as detected by in situ hybridization.

To understand regulation of the tracheo-bronchial mucin (TBM) gene expression we developed cDNA probes encoding TBM. We also raised antisera against mucin protein (deglycosylated and glycosylated), and developed immortalized tracheal epithelial cells which express mucin (at the RNA and protein level). TBM cDNA probes can detect TBM mRNA in situ in samples from dog and human primary tracheal epithelial cells and cell lines derived from them. For clinical application, conditions were optimized for detection of the TBM mRNA in human turbinal and nasal polyps and trachea of cystic fibrosis (CF) and non-CF subjects. Fixing and hybridization conditions were found to be critical for the optimum hybridization signal. Riboprobes proved to be better than cDNA or oligonucleotide probes. The application of these newly developed molecular tools in the genetic therapy of CF is discussed herein.

Animals↗
PubMed1996

Comparison of two substrates in determination of plasma cholinesterase activity & genetic variability.

Two substrates, acetyl thiocholine iodide ATCI and benzoylcholine chloride BCC were compared for the determination of plasma cholinesterase ChE levels and after incorporation of dibucaine and sodium fluoride in the assay, their usefulness in determining plasma ChE genetic variability was assessed in 64 healthy subjects. With both substrates, plasma ChE levels were found to be in the reference range. However, ATCI could detect only two variants with the usual phenotype UU in 60 of 64 [93.75%] subjects whereas with BCC 6 different groups could be determined. Though both substrates are of equal value in estimation of plasma ChE levels, BCC is definitely superior in determining its genetic variants.

Acetylthiocholine↗
PubMed1996

2,3,7,8-tetrachlorodibenzo-p-dioxin modulates expression of the prostaglandin G/H synthase-2 gene in rat thymocytes.

As an approach to understanding the molecular mechanism(s) of thymic gene expression mediated by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), we investigated the effect of TCDD on expression of prostaglandin G/H synthase-2 (PGHS-2) in rat thymocytes by reverse transcription-polymerase chain reaction. Incubation of thymocytes with increasing doses of TCDD resulted in inhibition of PGHS-2 gene expression in a concentration-dependent manner, with an IC50 of 10 nM. In contrast, TCDD had no appreciable effect on expression of glyceraldehyde phosphate dehydrogenase. Because the xenobiotic-responsive element is conserved in the PGHS-2 promoter from several animal species, it seems likely that inhibition of PGHS-2 expression by TCDD may occur at the level of transcription. To test this hypothesis in cultured thymocytes, we characterized the Ah receptor in the thymoma cell line WEHI 7.1. Reverse transcription-polymerase chain reaction experiments indicated that TCDD inhibited PGHS-2 expression in this cell line. Sucrose density gradient centrifugation experiments indicated that WEHI 7.1 cytosol exhibited 9 to 10S ligand-binding activity characteristic of the Ah receptor. The viability of WEHI 7.1 cells incubated with TCDD was comparable to that of control cells, whereas dexamethasone induced toxicity in a concentration-dependent manner. Transient transfection experiments using PGHS-2 promoter fragments ligated into a chloramphenicol acetyltransferase reporter plasmid suggested that TCDD inhibits PGHS-2 transcription, and deletion of the xenobiotic-responsive element failed to exhibit this repression. These results demonstrate that TCDD is a potent inhibitor of PGHS-2 gene expression, and they represent the first mechanistic evidence for TCDD-dependent inhibition of transcription in thymocytes.

Animals↗
PubMed1996

Plasma vitamin A, zinc and selenium concentrations in children with acute and persistent diarrhoea.

Plasma zinc, selenium, and vitamin A concentrations in 25 children with persistent diarrhoea (PD) and acute diarrhoea (AD) were determined and compared with 25 age-matched control children. Plasma retinol concentrations (PRC) and plasma zinc concentrations (PZC) (3.92 micrograms/dL and 79.4 micrograms/dL respectively) were found to be significantly lower (p < 0.05) in children with PD. PZCs were also significantly reduced (p < 0.05) in children with AD. However, reduction in PZC was more in PD than that in AD. There was no significant difference in PSC in children with either types of diarrhoea as compared with the control group. The results of the study showed that there was deficiency of vitamin A and zinc in diarrhoeas which needs to be correlated for proper nutritional management.

Acute Disease↗
PubMed1995

Primary structure of the canine U1 snRNA.

The nucleotide (nt) sequence of the canine U1 snRNA was determined. It exhibited significant homology (90-98%) with known U1 sequences. The RNA can be folded according to the secondary structure previously proposed for the U1 snRNA. It contained the conserved sequence UUACCUG in loop A (nt 6-12), required for the recognition of the 5' splice site, and the sequence UGCACU in loop B (nt 68-73), required for recognition of the U1-70K protein. The U1 snRNA was localized in the nucleus and its transcription was sensitive to alpha-amanitin, suggesting that it is transcribed by RNA polymerase II. Southern analysis revealed that the canine genome possesses 5-10 copies of U1 snRNA-encoding genes.

Animals↗
PubMed1995

51Cr-labelled ethylenediaminetetraacetic acid and D-xylose absorption test in Escherichia coli enterotoxin-induced diarrhoea in mice.

BACKGROUND AND METHODS: D-Xylose and 51Cr-labelled ethylenediaminetetraacetic acid (EDTA) absorption tests were evaluated in control and Escherichia coli heat-stable, heat-labile, and both heat-stable and heat-labile enterotoxin-induced diarrhoea in mice. RESULTS: The absorption of D-xylose was greatest 45 min after its ingestion in both control and enterotoxin-treated groups. There was a significant (p < 0.01) decrease in xylose absorption at 45 min in all the three experimental groups as compared with the control group, which may be due to change in membrane fluidity and hence change in membrane permeability, whereas 51Cr-EDTA absorption was at its maximum 15 min after its ingestion in both control and experimental groups. And there was a significant (p < 0.05) increase in 51Cr-EDTA absorption in enterotoxin-treated groups as compared with the control group. CONCLUSIONS: These results suggest a change in tight junctional permeability which may be due to change in the cytoskeletal microfilaments. Both these tests hence show that E. coli enterotoxin-induced diarrhoea results in absorption and permeability changes.

Animals↗
PubMed1995

Biophysical profile of blood pressure in school children.

The study was conducted in an industrial and prosperous city of Punjab to evaluate the biophysical profile of blood pressure (BP) in apparently healthy school children. A total of 2560 children between the ages of 5-15 years were enrolled. Their age, religion, dietary and family history were recorded. Weight and height of all children were measured and body mass index (BMI) calculated. A value of 2.26 or more was taken as obesity. BP measurements were made as per recommendations of the American Heart Association. Systolic as well as diastolic BP increased with age in both sexes, correlation coefficients being 0.59 and 0.6, respectively. A statistically significant linear relationship between BP and weight and height was noted. Children with BMI of > 2.26 had a significantly higher BP (P < 0.01). The mean BP did not vary among different religions. The BP of vegetarians and also non- vegetarians also did not differ. A family history of hypertension was associated significantly with elevated BP (p < 0.01). It is concluded that obesity and a family history of hypertension in children are associated with elevated BP and such children may be at risk for developing hypertension at a later date. They should be followed up and considered for modification of risk factors.

Adolescent↗
PubMed1995

Antenatal period: an educational opportunity.

In this study the impact of an educational programme during antenatal period was evaluated. Pregnant women attending the antenatal clinic formed the study material. The first 100 mothers who were not given health education served as controls. The subsequent 201 cases constituted the study group and were given health education on certain aspects of maternal and child care. The control and study groups were well matched for age, parity, education, income and number of antenatal visits. The results indicated that the mothers in study group gained statistically significant knowledge regarding the purpose of antenatal care, hematinics and tetanus toxoid vaccination. The awareness regarding breast feeding and its advantages also increased significantly in the study group. The knowledge about individual vaccine especially measles and DPT was poor which increased significantly after the educational intervention in the study group. It is recommended that the antenatal period should be optimally utilized to impart health education on the various aspects of maternal and child health.

Case-Control Studies↗
PubMed1995

Attitudes of future mothers regarding infant feeding.

Attitudes of 2500 urban college girls from Ludhiana city, aged 15 to 20 years were assessed towards infant feeding using a standardized Likert Scale (r = 0.82). The influence of subject opted in college and the presence of a child less than 3 years in the family on the attitudes was also studied. The overall attitudes of the whole group towards infant feeding were negative. Science students had better attitude scores on most of the themes tested (p < 0.05). The presence of a child aged less than 3 years in the family had no significant influence on the attitudes of girls towards infant feeding. Most of the girls were desirous of studying the concepts of infant feeding in their college curriculum. It is recommended that efforts should be made to incorporate these concepts in the formal educational curriculum so that girls grow up with positive attitude towards infant feeding.

Adolescent↗
PubMed1995

Predictive utility of clinical and stool parameters in bacterial diarrhoea in children.

A prospective one year study was conducted on children between the ages of 1 month to 5 years hospitalised in the pediatric ward of Christian Medical College, Ludhiana, with the aim of determining the predictive utility of certain clinical and stool parameters in diagnosing bacterial diarrhoea. Among the 204 children enrolled in the study, fever was observed in 40% in both the culture positive and negative groups. Clinical features such as abdominal distension, vomiting and oliguria although had low positive predictive values, their negative predictive values were high. Among the stool parameters, watery consistency and pus cells > 5 HPF were significantly more often observed in culture positive cases. The presence of mucus and pus cells > 5 HPF had good sensitivity (70-80%) but poor specificity (27-40%), while the reverse was true of blood (sensitivity 23%, specificity 89%). Again the positive predictive values were uniformly low while the negative ones were high. In conclusion the clinical and stool parameters were found to be more useful by their absence than by their presence in excluding a positive stool culture.

Bacterial Infections↗
PubMed1995

Substituted quinazolinones and their anti-inflammatory activity.

6-Substituted-2-alkyl-3-(4-aminobenzene sulphonamido)quinazolin-4-(3H) ones (1a-b) were converted either to 2-methyl 3-[(N-aminoacylbenzoate)benzenes sulphonamido]quinazolin-4 (3H) ones (2a-c) or to 6-substituted-2-alkyl-3-(N-arylidene benzene sulphonamido)-quinazolin-4- (3H) ones (3a-h). 3 on reduction, yielded 6-substituted-2-alkyl-3-(N-arylmethyl sulphonamido)-quinazolin-4(3H)ones (4a-l). Reaction of 3a-h with thioglycolic acid afforded 2-alkyl-3-[4'-(5'-arylthiazolidin-3'-one)benzene sulphonamido]quinazolin-4(3H)ones (5a-h). All these compounds were evaluated for their anti-inflammatory activity against carrageenin induced rat's paw oedema. Active compounds were also evaluated for ulcerogenic liability and ALD50 values.

Animals↗