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Biomedical subjects

M Verma

Publications and source records attributed to M Verma.

At least 37 records · Page 2Linked to original sources

Search for a novel killer toxin in yeast Pichia pastoris.

Certain yeast strains secrete a protein toxin, which inhibits the growth of sensitive pathogens and yeasts. Studies have shown that production of the toxin is dependent on presence of linear, double-stranded DNA plasmids in the killer yeasts. In the yeast Pichia pastoris, two linear double-stranded DNA plasmids have been identified. In the present study, the search for toxin-producing capability in P. pastoris has been conducted. No killer activity could be detected when 14 different indicator strains were tested.

Blastomyces↗

Aspirin esterases in North-West Indians: the influence of age and nutrition.

OBJECTIVE: To determine the activity of aspirin esterases in North-West Indian population and to find the effect of age and nutrition on it. SUBJECTS, MATERIAL AND METHODS: The serum albumin, plasma cholinesterase (PChE), aspirin esterase (ASPES) and phenyl acetate esterase (PAE) were determined in 175 subjects: young (< 40 years) and healthy (BMI > 19) = 74; elderly (> 50 years) and healthy (BMI > 19) = 32; young (< 40 years) and emaciated (BMI < 19) = 44; elderly (> 50 years) and emaciated (BMI < 19) = 25). RESULTS: The serum albumin levels significantly decreased with increase in age (r = -0.384, p < 0.01) and with decrease in body mass index (r = 0.457, p < 0.01). When the activity of esterases in four groups was compared, the PAE activity was not found to be affected by age or nutrition and the ASPES and PChE activity were significantly lower only in elderly emaciated (p < 0.01). CONCLUSION: As elderly emaciated have decreased serum albumin, ASPES and PChE activity, they may need a lower dose of aspirin to achieve the desired antiplatelet and analgesic effect. The young emaciated subjects, in spite of their lower serum albumin levels, may not require a lower dose of aspirin.

Adult↗

Ciprofloxacin treatment does not influence course or relapse rate of reactive arthritis and anterior uveitis.

OBJECTIVE: To assess the efficacy of ciprofloxacin in the treatment of reactive arthritis (ReA) and anterior uveitis (AU) in a double-blind, randomized, placebo-controlled trial. METHODS: Seventy-two patients participated in this study, 56 with ReA and 42 with AU (26 patients had both ReA and AU). Ciprofloxacin (750 mg twice a day) was administered for 12 months with a 12-month followup. End points of the study included time to disease relapse and measures of disease severity. RESULTS: There was no difference between groups in time to disease relapse, joint inflammation, number of joints and enthesis involved in patients with ReA, or signs and symptoms of AU. CONCLUSION: Long-term treatment of ReA and AU with ciprofloxacin made no statistically significant difference to the natural history of these diseases or their severity.

Anti-Infective Agents↗

CFTR antisense phosphorothioate oligodeoxynucleotides (S-ODns) induce tracheo-bronchial mucin (TBM) mRNA expression in human airway mucosa.

Mucus hypersecretion is a critical component of cystic fibrosis (CF) pathogenesis. The effects of dysfunction of the cystic fibrosis transmembrane regulator (CFTR) on mucin expression were examined using the tracheo-bronchial mucin (TBM) gene as an indicator. TBM mRNA expression was assessed in a human bronchial epithelial cell line (HBE1) and human nasal mucosal explants in vitro. Antisense phosphorothioate oligodeoxynucleotides (S-ODN) to TBM suppressed baseline expression of TBM mRNA in both systems, but had no effect on glyceraldehyde phosphate dehydrogenase mRNA (GAPDH) expression. Sense and missense (multiple scrambled control oligonucleotides) S-ODNs had no effect. 8Br-cAMP and PGE1 significantly elevated TBM mRNA expression. These increases were also specifically inhibited by the antisense S-ODNs. In order to induce a CF-like state, S-ODN to CFTR were added to explants. Antisense CFTR S-ODNs were anticipated to reduce the expression of cellular CFTR protein, and the level of CFTR function. Antisense, but not sense or missense, CFTR S-ODN significantly increased TBM mRNA expression. These data suggest that mucin hypersecretion in CF may be a direct consequence of CFTR dysfunction; the specific mechanism through which this effect is mediated is not known.

Cells, Cultured↗

Paraoxonase (PON1) polymorphism & its relation with lipids in north west Indian Punjabis.

We investigated 190 healthy, unrelated and randomly selected, north-west Indian Punjabis (M:102; F:88) for paraoxonase (PON1) polymorphism by dual substrate method and also determined lipid variables i.e., total cholesterol (TC), high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL) and triglycerides (TG) in them to determine any relationship between PON1 activity, PON1 phenotypes and lipids. The basal plasma paraoxonase (PON) activity, and PON activity in presence of 1 Mol NaCl (salt activated paraoxonase i.e., SAP) were estimated by using paraoxon as substrate whereas the, phenyl acetate esterase (A) activity was estimated by using phenylacetate as substrate. Based on the ratio of SAP/A activity, three distinct phenotypes of PON1 could be determined with gene frequencies of PON*A (low activity) and PON*B (high activity) allele being 0.847 and 0.153 respectively. In the whole population on partial correlation after normalising the variables and after adjusting the lipids for age and body mass index (BMI), a significant negative correlation was observed between SAP/A ratio and TC (r = -0.290; P < 0.01) and LDL (r = -0.154; P < 0.05). However, on analysis of covariance (ANCOVA) after normalizing the lipid variables and adjusting these for age and body mass index (BMI), no significant difference could be observed in lipid profile of these three phenotypes. The lack of a significant relationship between lipids and PON1 phenotypes, suggests that PON phenotype does not significantly influence the lipid profile in north-west Indian Punjabis. However, a significant negative correlation between the PON activity and TC and LDL suggests that low PON activity could be a risk factor for atherosclerosis in these subjects.

Adult↗

MUC1 upregulation by ethanol.

MUC1 is a glycoprotein and its expression is altered in breast cancer. Mucin protects epithelia from the external hostile environment. The expression of mucin changes when epithelia come in contact with toxic agents such as ethanol. Previously, we characterized the expression and regulation of tracheo-bronchial mucin (TBM) gene. In the present study, we studied the effect of ethanol on the gene encoding mammary gland mucin MUC1 and observed that ethanol regulates MUC1 expression at the transcription level. Ethanol enhanced the expression of MUC1 mRNA in a dose- and time-dependent manner in MCF-7 cells. At 100 mM concentration (a concentration reported to be present in alcoholics), ethanol induced a three to five-fold increase in mucin transcription as determined by nuclear run on analysis. This concentration of ethanol does not affect the half-life of MUC1 mRNA.

Adenocarcinoma↗

Development and characterization of new immortalized human breast cell lines

New human breast cell lines were developed from metastatic breast cancer tissues and normal breast tissues. Primary cultures were initiated from cellular outgrowths of explanted tissues or from mechanically isolated cells in two serum-free media. Cell cultures derived from both cancer and normal tissues were immortalized with pRSV-T plasmid to generate permanent breast cell lines that exhibited an epithelial morphology. Cell lines generated in this study were characterized with respect to morphology, growth rate, karyotype, presence of specific genes, and the expression of epithelial and breast markers. The cell lines expressed the epithelial cell markers, cytokeratins 8 and 18, and retained the capacity to produce human milk fat globulin. They also express the BRCA-1, erbB2, and EGF receptor genes and possess the H-ras, K-ras, and p53 genes. Preliminary data showed that one of the new cancer cell lines was highly sensitive to the cytotoxic action of taxol. It is envisioned that the new breast cell lines will be useful as targets for identification of therapeutic agents against breast cancer and as models for carcinogenesis studies.

Journal Article↗

Establishment and characterization of immortalized human cell lines from prostatic carcinoma and benign prostatic hyperplasia.

New human prostate cell lines were developed from prostatic carcinoma (BRF-41T) and BPH (BRF-55T). Primary cultures were initiated from cellular outgrowths of explanted tissues. A serum-free medium, BRFF-HPC1, was developed for growing human prostatic cancer cells. Cell strains were immortalized with pRSV-T plasmid to generate permanent cell lines that exhibited an epithelial morphology. Both cell lines expressed the epithelial cell markers, cytokeratins 8 and 18 as well as the prostatic marker, PSA, and the androgen receptor gene. They possess the H-ras, K-ras, and p53 genes. We hope that these new human prostatic cell lines will be useful as in vitro models for cancer research.

Cell Culture Techniques↗

Prevalence of anemia among urban school children of Punjab.

OBJECTIVE: To assess the prevalence of anemia among urban school children of Punjab. DESIGN: Cross sectional study. SETTING: Urban schools of Ludhiana, Punjab. SUBJECTS: Two thousand school children of 5-15 years age. METHODS: Relevant history was taken and a complete physical examination done in all the children. Hemoglobin was estimated using cynmethemoglobin method and peripheral blood smears were also examined. Anemia was diagnosed when hemoglobin was less than 11 g/dl for children of 5-6 years age and 12 g/dl for more than 6 years age. RESULTS: Overall prevalence of anemia was 51.5%. Girls had a significantly higher prevalence of anemia except at 5 years and 10-12 years age. More menarcheal girls were anemic as compared to non-menarcheal ones. The prevalence of anemia was high (38%) even in higher socioeconomic groups. Nearly half (47.6%) of well nourished children were anemic. The mean Hb also was lower than expected normal values in both nutritional groups. Compared to non-vegetarians (38%), more vegetarians (65.9%) were anemic. The commonest blood picture seen was microcytic hypochromic (55.4%). CONCLUSIONS: The present study revealed a high prevalence of anemia among healthy urban school children of higher socio-economic classes. Vegetarians and girls, especially after menarche were more at risk to develop anemia.

Adolescent↗

Paraoxonase polymorphism in northwestern Indians.

OBJECTIVE: The aim of this study was to distinguish paraoxonase phenotypes by dual substrate method. SUBJECTS AND METHODS: 166 unrelated healthy northwestern Indian Punjabis were studied for basal paraoxonase activity (P), in presence of 1 M NaCl (salt-activated) using paraoxon as substrate and arylesterase activity (A) using phenylacetate as substrate. RESULTS: The mean (+ SD) activity of these in population was 70.43 (+/-30.41), 103.11 (+/-42.87) nmol/min/ml plasma and 36.81 (+/- 9.87) micromol/min/ml plasma, respectively, with no significant difference in sexes. Based on the ratio of SAP activity to A activity, 3 distinct phenotypes could be determined with gene frequencies of PON*A and PON*B being 0.845 and 0.155, respectively. Between P and A, and SAP and A in both A and AB phenotypes, significant correlation was observed (A = r 0.34 and 0.62, p < 0.01; B = r 0.31 and 0.81, p > 0.01), suggesting that both enzymes belong to the same group. CONCLUSIONS: Though northwestern Indian Punjabis have trimodal paraoxonase activity, the majority is either low homozygous (AA) or heterozygous (AB) type with enzyme activity lower than Europids.

Adult↗

Enolase activity in chicken embryo primary retina cells is not affected by exposure to a 60-Hz electric field.

This study was to determine the extent of alteration of enolase specific activities in chicken embryo retina primary cells in culture when exposed to an ELF (extremely low frequency) electric field at 60 Hz. Results showed no alteration of enolase activity and enolase mRNA levels. In this study, sham vs. control experiments were also conducted to neutralize ambient AC magnetic fields, stray magnetic fields and variations in field uniformity. Under similar conditions, the specific activity of enolase is decreased in neuroblastoma cell line (NG108). It is apparent from this study that primary cells either are not affected by these exposure conditions or the effect is transient and warrants no damage.

Animals↗

Calcium and protein kinase C play a significant role in response to Shigella toxin in rabbit ileum both in vivo and in vitro.

The role of second messengers in Shigella toxin (STx) induced fluid secretion in rabbit ileum was evaluated. In vivo and in vitro studies were carried out in presence or absence of following modulators: Ca2+ ionophore A23187 (15 microM), l-verapamil (200 microM), phorbol-12-myristate-13-acetate (PMA, 200 ng), 1-(5-isoquinolinyl-sulphonyl)-2-methyl-piperazine (H-7, 15 microg) and indomethacin (20 microM). In in vivo studies, the fluid accumulation into rabbit ileal loops in response to STx was measured in presence or absence of these modulators. In in vitro studies, unidirectional fluxes of Na+ and Cl- were carried out in presence or absence of these modulators. The addition of Ca2+ ionophore A23187 along with STx further increases the amount of fluid already induced by STx. Whereas the presence of l-verapamil along with STx did not decrease the amount of fluid induced by STx. In vitro findings were in consonance with the in vivo studies. A significant increase in inositol triphosphate (IP3) levels was observed in enterocytes isolated from STx treated rabbit ileum. The addition of PMA into rabbit ileal loops in presence of STx mimicked the effect of STx while the presence of H-7 reversed the secretion caused by STx to absorption. Similar results were obtained while determining unidirectional fluxes of Na+ and Cl- in presence of PMA and also with H-7. A significant increase in PKC levels was observed in the membrane fraction of enterocytes isolated from STx treated rabbit ileum as compared to control. Further a marked decrease in PKC levels was observed in the presence of H-7 in membrane fraction of enterocytes isolated from STx treated rabbit ileum. The addition of indomethacin into rabbit ileal loops reversed the secretion (caused by STx) to absorption. In vitro findings were in consonance with in vivo studies. Besides, there was a significant increase in PG-E levels in enterocytes isolated from STx treated rabbit ileum as compared to control. These findings suggested that STx induced enteritis involves the role of PKC, intracellular calcium stores and prostaglandins. The extracellular calcium pool probably does not play a significant role in this process.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Transcriptional regulation of tracheo-bronchial mucin (TBM) gene by ethanol.

The epithelia of the respiratory tract are protected by a mucin glycoprotein. The expression of mucin changes when epithelia come in contact with toxic agents such as ethanol. Previously, we have identified and characterized the expression of a tracheo-bronchial mucin (TBM) gene. In the present study, we observed that ethanol regulates TBM expression at the transcription level. Ethanol enhanced the expression of TBM mRNA in a dose- and time-dependent manner in HBE1 cells. At 100 mM concentration (a concentration reported to be present in alcoholics), ethanol induced an eight-fold increase in TBM transcription as determined by reporter gene expression analysis.

Bronchi↗

Dynamics of conjunctival impression cytologic changes after vitamin A supplementation.

To investigate the chronological changes in conjunctival epithelium after supplementation with a massive oral dose of vitamin A, conjunctival impression cytology (CIC) with transfer was carried out repeatedly among 200 children aged 6-120 months in a randomized, double-blind, placebo-controlled study in Chandigarh (India). Significant conversion to normal CIC started 71-80 d after vitamin A supplementation and by 101-110 d conversion had taken place in all children. Compared with the placebo group, plasma retinol concentration at 100 d post-supplement was found to be significantly higher in the vitamin A-supplemented group P = 0.04. This study demonstrates that CIC responds to a massive oral dose of vitamin A 3-4 months after supplementation. These findings should guide future studies and evaluations in which CIC is used to assess response to vitamin A interventions.

Administration, Oral↗

RNA-RNA interaction and gene splicing.

The precise excision of intervening sequences during RNA splicing is an interesting example of the high degree of specificity involved in biosynthesis processes. Self-splicing RNA precursors achieve this specificity primarily through intramolecular interactions whereas all other types of RNA splicing requires interaction between cellular factors and specific recognition signals in the RNA precursor. About twelve years ago, the in vitro splicing system was developed and a general scheme of the pre-mRNA was proposed (Hernandez and Keller, 1983; Krainer et al., 1984; Lin et al., 1985; Padgett et al., 1984; Ruskin et al., 1984). A fundamental question in the splicing field is how the 5' and 3' splice sites are recognized and paired during the splicing reaction. Recent work in the splicing field has established that a network of RNA interactions may form the structural foundation of the spliceosomes. Possible solutions to many unsolved puzzles are getting attention. RNA-RNA interactions now appear to underlie many aspects of substrate recognition, reaction partner juxtaposition and catalysis. In this article we have presented the latest mechanisms involved in the pre-mRNA splicing and their implication in applied research including cancer.

RNA↗

Red cell acetyl cholinesterase and plasma cholinesterase activity and genetic variants of plasma cholinesterase in northwest Indian adults.

The plasma cholinesterase (PChE) and red cell acetyl cholinesterase (AChE) activities are indicators of exposure to organophosphates. We studied their distribution in unexposed Northwest Indian adults by measuring them in 120 men and 111 women by Ellman's and Kalow's method, respectively. We also determined genetic variability of plasma cholinesterase in 193 subjects (male = 111, female = 82). The mean +/- (SD) AChE levels in population, men and women, were 34.97 +/- 13.66, 35.05 +/- 12.42, 34.88 +/- 14.89 nmol/mg Hb/min, whereas PChE was 0.448 +/- 0.173, 0.435 +/- 0.163, 0.462 +/- 0.183 ku/l, respectively. When compared for sex, no significant difference could be found for red cell AChE and PChE activity. However, on 2-way analysis of variance (ANOVA) adjusted for age classification, the levels of both AChE and PChE were significantly higher in groups above the age of 30 years as compared to below 30 years (t = 3.08, p < 0.01, t = 2.82, p < 0.05), respectively. Seven genetic variants of PChE could be detected in males, whereas in females 6 genetic variants were found.

Acetylcholinesterase↗