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Biomedical subjects

M Sharma

Publications and source records attributed to M Sharma.

At least 109 records · Page 6Linked to original sources

Epidemiological determinants of burns in paediatric and adolescent patients from a centre in western India.

A 1-year prospective study of 112 burn patients up to 19 years of age aimed to identify and study the determinants and mortality experiences in these burn patients and found flame burns to be the commonest, followed by scalds and electric burns. All burns other than those caused by electricity were commoner in females, more so between 15 and 19 years of age. Burns were more frequent in winter and 85 per cent of them were domestic. Nearly all burns took place during day time with a higher incidence between 06.00-09.00 h and 14.00-21.00 h. The patient fatality rate (41.1 per cent) was associated with total burn surface area. Referral time-lag was an important determinant of mortality especially in less severe burns. As would be expected, hospital stay was significantly longer in survivors.

Accidents

Neuroimaging of meningeal disease.

In the past two decades, the advent of CT and MRI has made a considerable impact on the evaluation of meningeal diseases, conditions once regulated to cytological, histopathological, or postmortem analyses alone. This article reviews the imaging findings in various meningeal processes with particular attention to the anatomic definition of the meningeal layers and their relationship to the development of meningeal pathology and consequent imaging characteristics.

Central Nervous System Diseases

Characterization of monoclonal antibodies to conserved antigens of Entamoeba histolytica and E. dispar and development of a stool ELISA.

Eight murine monoclonal antibodies (MAbs) were generated against Entamoeba histolytica NIH:200. All MAbs reacted with three axenic strains of E. histolytica, NIH:200, HM-1: IMMS, and SAW 1734R clAR, but not with enteric bacteria or Giardia lamblia. Five of the MAbs reacted with low-molecular-weight, periodate-sensitive antigens of 14-21 kD, while one (AB31) reacted with high-molecular-weight, 90 to 200-kD protein determinants. MAb PC14 appeared to be specific for antigen in its native state. Another MAb (BB12) agglutinated live trophozoites and caused membrane fluorescence in contrast to the five other MAbs tested. Although BB12 reacted with the same 14 to 21-kD band on Western blot as AC55, the latter reacted with different cytoplasmic epitopes. All the MAbs reacted to five pathogenic (E. histolytica) and six nonpathogenic (E. dispar) clinical isolates. These MAbs may be helpful for studying conserved antigens of E. histolytica and were used to develop a sandwich ELISA for the diagnosis of intestinal amoebiasis. The assay was sensitive to 60 ng of E. histolytica antigen. A comparative study of microscopic examination of stool samples and the sandwich ELISA was conducted on 102 samples from patients with gastrointestinal complaints. The ELISA could detect all microscopically positive samples with a sensitivity of 100% and specificity of 93%. A sandwich ELISA using a monoclonal antibody to conserved antigens of E. histolytica has the potential to be a reliable method for the diagnosis of intestinal amoebiasis.

Agglutination Tests

Primary radiation therapy to T1 and T2 breast cancer following conservative surgery. Which patients should be boosted?

We reviewed 199 radiated patients at our institution (201 breasts treated) and its affiliates treated between 1978 and 1989. Of these, 157 were T1 and T2 invasive breast carcinoma. Our intent was to retrospectively compare the results of those who received standard doses of 4,500 to 5,000 cGy to the breast to those that received an additional boost to the surgical bed to a dose totaling at least 5,500 cGy. There were a total of 5 local recurrences in 159 treated breasts. (The mean follow-up time was 36 months.) Of our T1 and T2 patients with clear resection margins that were boosted, there was 1 local recurrence in 28 treated breasts. There was 1 local recurrence in the nonboosted group of 68 patients. Except for one patient, all patients with positive margins were boosted. There were 2 local recurrences in the 23 T1 and T2 breasts with positive margins that were boosted. Of the patients with uncertain margins who were not boosted, there was one local recurrence in 20 treated breasts. Of those with uncertain margins that were boosted, there were no local recurrences in 19 treated breasts. From our results, it would appear that a boost to the primary site is unnecessary if the margins of resection are negative (by either inking or if it is clearly stated in the pathology report). In those patients with uncertain margins, most were done in the years before margins were routinely inked, but generous excisional biopsies were usually done. In this latter group of patients, there also was no added benefit to boosting.

Adult

Purification and characterization of the SegA protein of bacteriophage T4, an endonuclease related to proteins encoded by group I introns.

Although not encoded by an intron, the bacteriophage T4 SegA protein shares common amino acid motifs with a family of proteins found within mobile group I introns present in fungi and phage. Each of these intron-encoded proteins is thought to initiate the homing of its own intron by cleaving the intronless DNA at or near the site of insertion. Previously, we have found that SegA also cleaves DNA. In this report, we have purified the SegA protein and characterized this endonuclease activity extensively. SegA protein cleaved circular and linear plasmids, DNA containing unmodified cytosines, and wild-type T4 DNA containing hydroxymethylated, glucosylated cytosines. In all cases, certain sites on the DNA were highly preferred for cleavage, but with increasing protein concentration or time of incubation, cleavage occurred at many sites. SegA cleaving activity was stimulated by the presence of ATP or ATP gamma S. Sequence analysis of three highly preferred cleavage sites did not reveal a simple consensus sequence, suggesting that even among highly preferred sites, SegA tolerates many different sequences. A T4 segA amber mutant that we constructed had no phenotype, and PCR analyses indicated that several T-even-related phages lack the segA gene. Taken together, our results show that SegA is an endonuclease with a hierarchy of site specificity, and these results are consistent with the insertion of segA DNA into the T4 genome some time after the divergence of the closely consistent with the insertion of segA DNA into the T4 genome some time after the divergence of the closely related T-even phages.

Amino Acid Sequence

MC540 induced photosensitization of glioma & neuroblastoma cells.

Binding and photodynamic action of merocyanine 540 (MC540) has been studied in glioma (U-87MG) and neuroblastoma (Neuro 2A) cells as a function of dye concentration, incubation time of cells with MC540 and growth phase of cells. In the plateau phase, U-87MG cells accumulated more MC540 as compared to exponentially growing cells, whereas in Neuro 2A cells the opposite effect was observed. Exponentially growing U-87MG cells were more photosensitive than plateau phase cells. However, the photosensitivity of Neuro 2A cells was not dependent on the growth phase. Thus, MC540 mediated photosensitization may be useful for photodynamic therapy of brain tumours.

Fluorescent Dyes

A quantitative microbiological study of bacterial vaginosis.

A quantitative microbial study of vaginal secretions from 100 women with bacterial vaginosis (BV) and 50 healthy women matched for age, was undertaken. This was aimed at comparing the bacterial flora colonising the vagina of sexually active asymptomatic women with those with BV and correlating bacterial counts with presence of symptoms. Of the total 475 bacteria isolated anaerobes were predominant among controls and among patients aerobes and anaerobes had equal prevalences. The increased viable counts of all anaerobes (> 10(7) cfu/g) were related with symptoms. Mobiluncus mulieris could be isolated from one patient and Chlamydia trachomatis antigen was absent in all samples tested. Bacteroides, G. vaginalis and Ureaplasma urealyticum were found to be the most commonly occurring group in women with BV. Lactobacillus, and probably Coryneforms have a protective role in health, preventing disease.

Adolescent

Effect of D-penicillamine on lymphocyte subsets: correlation with clinical response in rheumatoid arthritis.

We studied the effects of D-penicillamine (DP) on the clinical response, immunoinflammatory parameters and the lymphocyte subsets in 46 patients with rheumatoid arthritis (RA). Patients were evaluated before the start of the drug and then at 3 and 9 months during the follow up. 38 of 46 (82.6%) patients could continue DP treatment for over 9 months, while in 8 the drug was withdrawn due to adverse effects. Improvement in the various disease activity indices of more than 50% (responders) was seen in 25 of 38 (65.8%) patients. Responders showed a significant decrease in the serum IgA and IgM at 9 months, and in IgM only at 3 months. The serum levels of C3 and C4 did not show any significant change. Serum levels of C-reactive protein and rheumatoid factor (RF) showed a significant decrease at 3 and 9 months. A significant decrease in CD3+ and CD4+ lymphocytes along with a fall in CD4+/CD8+ lymphocyte ratio was also seen in responders at 3 and 9 months, compared to the baseline. Our results suggest that DP may have immunomodulatory action in RA.

Adolescent

Mode, presentation, CT findings and outcome of pediatric head injury.

Twenty five to thirty per cent of all patients admitted with head injuries to any large hospital are children. Head injuries in children differ in several ways from those seen in the adult population. A standard management protocol is used in a service hospital and CT and neurosurgical consultation/transfer is based on specific criteria. The records of 312 patients admitted with head injury to Command Hospital (WC), Chandimandir were retrospectively analysed to compare the mode of injury, initial presentation, CT findings and the outcome of management between those above and those below the age of 10 years. Eighty seven (27.8%) of the 312 patients were children and 71% of them had sustained the injury due to fall from a height. One third of children were brought with vomiting and drowsiness and 9.2% with seizures, but lateralizing signs were found in only 5.7%. Nineteen children underwent a CT of the head and of these 74% had only cerebral edema. Only 4 of the 87 children were transferred to a neurosurgical unit. The mortality rate was lower in children (5.7%) compared to the older age group (11.6%) but the difference was not statistically significant. However, a significantly higher number (80.5%) of children recovered without any residual deficit. Majority of our children can be safely managed in a general surgical unit provided a multidisciplinary approach is used and facilities of CT are readily available, since very few require active neurosurgical intervention.

Accidental Falls

Phytotoxicological evaluation of Tagetes erectes on aquatic stages of Anopheles stephensi.

Petroleum ether (5%) in ethyl acetate fraction of Tagetes erectes showed toxic activity against second and fourth instar larvae of Anopheles stephensi below 100 ppm concentrations. The LC50 values for second and fourth instar larvae were calculated to be 43 and 58 ppm respectively. The second instar larvae were more susceptible than the fourth instar larvae. The extract did not affect the larval developmental period. The growth index of treated mosquitoes was significant in comparison with that of the control and untreated sets (p < 0.01). Treatment with the extract had a significant effect on the mortality and reduction in the adult emergence of the vector.

Animals

Effects of carbon dioxide-induced anesthesia on cholinergic parameters in rat brain.

We report that acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) activities in rat brain were virtually identical whether the rat was anesthetized with carbon dioxide (CO2) before decapitation or decapitated without prior sedation. The AChE and ChAT activities were measured in three brain regions: the hippocampus, cerebral cortex, and cerebellum. Enzyme activities varied significantly by brain region, with the highest values in the hippocampus and the lowest values in the cerebellum. Enzyme activities, however, did not vary with the method of euthanasia, either CO2-induced anesthesia prior to decapitation or decapitation without anesthesia. These data suggest that CO2-induced anesthesia prior to decapitation does not alter activities of these cholinergic markers in rat hippocampus, cerebral cortex, and cerebellum. This method of euthanasia eliminates the need to capture a conscious animal, which reduces stress to the animal and the experimenter.

Acetylcholinesterase

Differential effects on spatial navigation of immunotoxin-induced cholinergic lesions of the medial septal area and nucleus basalis magnocellularis.

The effects on anatomy and behavior of a ribosomal inactivating protein (saporin) coupled to a monoclonal antibody against the low-affinity NGF receptor (NGFr) were examined. In adult rats, NGFr is expressed predominantly in cholinergic neurons of the medial septal area (MSA), diagonal band nuclei, and nucleus basalis magnocellularis (nBM), but also in noncholinergic cerebellar Purkinje cells. Rats with immunotoxin injections to the MSA, nBM, and lateral ventricle were compared to controls on a spatial and cued reference memory task in the Morris maze. Toxin injections to the MSA slightly impaired the initial, but not asymptotic, phase of spatial navigation. Injections to the nBM impaired all phases of spatial navigation. Cued navigation, however, was not affected in either the MSA or nBM group. The ventricular injections severely affected spatial and cued navigation. Acetylcholinesterase (AChE) histochemistry and NGFr and choline acetyltransferase immunohistochemistry revealed a loss of (1) almost all NGFr-positive cholinergic neurons in the MSA and AChE fibers in hippocampus (MSA group); (2) almost all NGFr neurons in the nBM, some in the MSA, most AChE fibers in neocortex and some in the hippocampus (nBM group), and (3) almost all NGFr neurons in the MSA and nBM and their corresponding hippocampal and cortical AChE fibers (ventricular group). Cholinergic nBM projections to the amygdala were largely preserved in all groups. The amount of cholinergic fiber loss in the cortex correlated modestly, but significantly, with the severity of impairment of the asymptotic phase of performance of the spatial task. An unambiguous interpretation of the anatomical locus of behavioral deficits was not possible because of damage to cholinergic striatal interneurons (nBM group) and to noncholinergic cerebellar Purkinje cells (ventricular group). These data suggest that the cholinergic cortical system is critical to the performance of this spatial memory task. Cholinergic denervation of the hippocampus alone, however, is not sufficient to impair markedly performance of this task.

Animals

Protective effect of Rubia cordifolia on lipid peroxide formation in isolated rat liver homogenate.

Rubia cordifolia Linn. (Rubiaceae) is an important component of the ayurvedic system of medicine. It has a variety of uses such as blood purifier, immunomodulant, antiinflammatory and anti-PAF. In this report the anti-peroxidative property of the solvent free alcoholic extract of R. cordifolia has been studied in rat liver homogenate. It prevents the cumene hydroperoxide induced malondialdehyde formation in the dose and time dependent manner. This effect is accompanied by the maintained reduced glutathione level even in the presence of above toxin.

Animals

Carriage of Gardnerella vaginalis in the urethra of Indian men.

A total of 310 consecutive sexually active men attending sexually transmitted disease clinic of the Nehru Hospital, Chandigarh, were studied to assess the prevalence of G vaginalis. Non-gonococcal urethritis (NGU) was diagnosed in 53, acute gonococcal urethritis (AGU) in 20, psychosomatic disorders in 50 and genital ulcer disease in 187 patients. G. vaginalis was isolated in 11 patients (3.5%) only. Of these 11 isolates, 4(7.5%) were from patients with NGU, 2(10.0%) from men diagnosed to have AGU, 3(6%) from the patients with psychosomatic disorders and 2 isolates (1.7%) from patients with genital ulcer disease.

Bacterial Infections

Effect of chronic cadmium exposure on glutathione S-transferase and glutathione peroxidase activities in rhesus monkey: the role of selenium.

The effect of cadmium (Cd) on the activity of glutathione S-transferase (GST) and glutathione peroxidase (GSH-Px) which play an important role in the detoxification of xenobiotics, was studied in the liver, kidney, heart and lung of Rhesus monkeys. Furthermore, the role of selenium (Se) in the modulation of Cd toxicity with respect to GST and GSH-Px was also evaluated. Cadmium exposure (5 mg Cd/kg body wt./day as CdCl2 for 10 weeks) to monkeys resulted in decreased GSH-Px activity in all four organs present in the order liver > kidney > heart > lung. Cadmium administration also resulted in a significant decrease in total GST activity present in the order liver > heart > kidney > lung, whereas a significant increase in the pi class GST activity was observed greatest in the heart followed by lung, kidney and liver. Oral administration of Se (0.5 mg Se/kg body wt./day as Na2SeO3 for 10 weeks) caused a significant increase in GSH-Px activity in the order liver > heart > kidney > lung. Selenium administration caused an increase in total GST activity in liver and lung but a decrease in kidney and heart. Simultaneous administration of Cd and Se resulted in an increase in total GST activity (except in lung) including the pi class activity as well as GSH-Px activity in all four tissues under study. Thus, the mechanism by which selenium decreases Cd toxicity in Rhesus monkeys, seems to rely on the protection of the enzyme systems GST and GSH-Px in the four organs, possibly by forming non-toxic cadmium selenide.

Animals

Fluorescence postlabeling assay of DNA damage induced by N-methyl-N-nitrosourea.

We have used a new technique to assay DNA adduct by combining enzymatic digestion of DNA to nucleotides and fluorescence postlabeling. The assay relies on the selectivity of nucleotide chromatography by high-performance liquid chromatography and the sensitivity of fluorescence detection. This report describes the fluorescence postlabeling assay of N7-(N7-methyldGuo) and O6-methyl-2'-deoxyguanosine (O6-methyldGuo) in calf thymus DNA exposed to N-methyl-N-nitrosourea. Using a conventional fluorescence detector, fluorescence postlabeling assay detected 1 modified nucleotide/10(6) normal nucleotides in 100 micrograms DNA. Laser induced fluorescence detection offers a linear response of the chromatographic signal from 10(-13)-10(-16) mol fluorescent nucleotides (correlation coefficient, 0.998) improving the detection limit of the assay to 1 modified nucleotide/10(8) normal nucleotides in 10 micrograms DNA. Fluorescence postlabeling analysis of the authentic markers shows that both N7- and O6-methyldGuo can be assayed with similar detection sensitivity. However, the inherent nature of the instability of N7-methyldGuo, even under physiological conditions, makes its quantitation difficult by any postlabeling technique. O6-MethyldGuo, on the other hand, can be detected with sufficient selectivity and sensitivity by a fluorescence postlabeling assay. Quantitative efficiency of enzymatic excision and chemical postlabeling of O6-methyldGuo were validated by fluorescence postlabeling analysis of synthetic model DNA. Fluorescence postlabeling assay complements 32P-postlabeling assay for O6-methyldGuo without requiring the handling and disposal of radiolabel. Fluorescence postlabeling assay has potential, therefore, to monitor the level of O6-alkyldGuo in human DNA exposed to both environmental and chemotherapeutic alkylating agents.

Animals

Fluorinated carbohydrates as potential plasma membrane modifiers. Synthesis of 3-deoxy-3-fluoro derivatives of 2-acetamido-2-deoxy-D-hexopyranoses.

Treatment of benzyl 2-acetamido-4,6-O-benzylidene-2-deoxy-alpha-D-allopyranoside with diethylaminosulfur trifluoride or of the 3-O-mesyl derivative with tetrabutylammonium fluoride gave the 2,3-unsaturated compound instead of the expected 3-deoxy-3-fluoro derivative. The latter was obtained when benzyl 2-acetamido-4,6-di-O-benzyl-2-deoxy-3-O-mesyl-alpha-D-allopyran oside was treated with potassium fluoride. Methyl 2-azido-4,6-O-benzylidene-2-deoxy-alpha-D-altropyranoside was converted into the 2-acetamido- and 2-phthalimido-3-O-mesyl derivatives; when treated with fluoride nucleophile, these gave only the 2,3-aziridine derivative. However, treatment of the 2-azido-2-deoxy derivative with diethylaminosulfur trifluoride gave methyl 2-azido-2,3-dideoxy-3-fluoro-alpha-D-mannopyranoside which, after reduction, deprotection, and acetylation, gave the acetylated derivative of methyl 2-acetamido-2,3-dideoxy-3-fluoro-alpha-D-mannopyranoside in excellent yield. These acetylated 3-fluoro derivatives exhibited inhibition of cell growth of murine L1210 leukemia cells in culture at micromolar concentrations.

Acetylglucosamine