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Biomedical subjects

P K Dutta

Publications and source records attributed to P K Dutta.

At least 19 recordsLinked to original sources

Isolation of pure compound R/J/3 from Pluchea indica (L.) Less. and its anti-amoebic activities against Entamoeba histolytica.

The plant Pluchea indica is known for its anti-inflammatory, anti-ulcer, anti-pyretic, hypoglycemic, diuretic and anti-microbial activities besides many other pharmacological activities. We have isolated and purified seven compounds from the methanolic root extract of this plant by column chromatography. The compounds were identified by spectroscopic analyses. The anti-amoebic activities of the pure compound R/J/3 was investigated against the HM1 strain of Entamoeba histolytica. The compound, R/J/3 showed the most pronounced anti-proliferative activity at a dose of 50 microg/ml. It also showed a marked activity on cell lysis of trophozoites, 4h after administration. The cell lytic activity was compared with metronidazole (5 microg/ml) as positive control.

Animals↗

A novel wavelet neural network based pathological stage detection technique for an oral precancerous condition.

AIM: To describe a novel neural network based oral precancer (oral submucous fibrosis; OSF) stage detection method. METHOD: The wavelet coefficients of transmission electron microscopy images of collagen fibres from normal oral submucosa and OSF tissues were used to choose the feature vector which, in turn, was used to train the artificial neural network. RESULTS: The trained network was able to classify normal and oral precancer stages (less advanced and advanced) after obtaining the image as an input. CONCLUSIONS: The results obtained from this proposed technique were promising and suggest that with further optimisation this method could be used to detect and stage OSF, and could be adapted for other conditions.

Algorithms↗

Studies on the production and purification of an antimicrobial compound and taxonomy of the producer isolated from the marine environment of the Sundarbans.

A microorganism isolated from the Sundarbans region of the Bay of Bengal, India, showed potent antimicrobial activity against gram-positive and gram-negative bacteria, molds, yeast and several multiple-drug-resistant (MDR) bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). The isolate grew in the presence of 20% (w/v) NaCl, antibiotic production being maximum with 5% (w/v) NaCl in the production medium. Natural seawater stimulated antibiotic biosynthesis. The absence of catabolite repression during the synthesis of the antimicrobial substance was demonstrated by the utilization of glucose by this isolate. The 16S rRNA gene of this aerobic, gram-positive, mycelium- and spore-forming microorganism was amplified, and molecular phylogenetic analysis of the DNA sequence showed less than 93% similarity with its closest relative, indicating differentiation at the genus level. The highly stable, active principle was purified by butyl acetate extraction and silica-gel chromatography and a single compound was found to posses the broad-spectrum activity. Molecular characterization showed that the active compound is a lipid. Bioreactor studies demonstrated that antibiotic production is strongly dependent on the scale of operation and there is a definite relation between the dissolved oxygen concentration, medium pH, glucose utilization, cell differentiation and antibiotic production. Maximum production in 30 h could be obtained by regulation of the medium pH in the alkaline range by a combination of controlled addition of NaOH, regulation of the air supply and changes in the reactor configuration. Considering all of the above evidences and based on comparison with the current literature, a novel antimicrobial appears to have been isolated.

Anti-Bacterial Agents↗

Distribution of virulence genes in Salmonella serovars isolated from man & animals.

BACKGROUND & OBJECTIVES: Pathogenesis of Salmonellosis depends upon a large number of factors controlled by an array of genes that synergise into the actual virulence of Salmonella. A study was undertaken to observe the distribution of three such genes, namely, Salmonella enterotoxin (stn), Salmonella Enteritidis fimbrial (sef and plasmid encoded fimbrial (pef genes, among different serovars of Salmonella enterica isolated from man and animals. METHODS: A total of 95 isolates belonging to S. Typhimurium (51), S. Enteritidis (36), S. Bareilly (3), and S. Paratyphi B (5) serovars were subjected to polymerase chain reaction (PCR) assay for the detection of stnl ssf and pef genes using their specific primers and the PCR products were analysed by 1 per cent agarose gel electrophoresis for the presence of the respective genes. RESULTS: Varying distribution pattern of these genes was observed amongst the isolates. While, stn was found in all the 95 strains, sef was found only among the S. Enteritidis isolates. The pef gene was found to be absent in 10 isolates including the three S. Bareilly isolates. INTERPRETATION & CONCLUSION: Findings indicated that the stn gene is widely distributed among Salmonella irrespective of the serovars and source of isolation. However, the sef gene appears to be serovar specific. Since the stn gene is found in all the isolates, it can be a viable target gene to explore the possibility of direct detection of Salmonella from samples from biological sources.

Adhesins, Bacterial↗

Enhanced frequency of micronuclei in individuals exposed to arsenic through drinking water in West Bengal, India.

In West Bengal, India arsenic in ground water has been found to be above the maximum permissible limit in seven districts covering an area of 37,493km2. In the present study, evaluation of the micronuclei (MN) formation in oral mucosa cells, urothelial cells and peripheral blood lymphocytes was carried out in the symptomatic individuals exposed to arsenic through drinking water. Forty five individuals with cutaneous signs of arsenicism from four affected districts (368.11 microg/l of As in drinking water) were considered as the exposed group and 21 healthy individuals with no symptoms of arsenic poisoning and residing in two unaffected districts (5.49 microg/l of As) were considered as controls. The exposed and control groups had similar age distribution and socioeconomic status. Standardised questionnaires were utilised and medical examination was conducted to ascertain exposure history, sociodemographic characteristics, diet, health, medication, addiction and chief symptoms in the study participants. Arsenic exposure was confirmed by measuring the arsenic content in the drinking water, nails, hair and urine samples from the volunteers. Arsenic contents in the urine, nail and hair in the exposed group were 24.45 microg/l, 12.58 and 6.97 microg/g, respectively which were significantly high in comparison to corresponding control group values of 4.88 microg/l, 0.51 and 0.34 microg/g, respectively. Exposed individuals showed a statistically significant increase in the frequency of MN in oral mucosa, urothelial cells and lymphocytes (5.15, 5.74 and 6.39/1000 cells, respectively) when compared with the controls (0.77, 0.56 and 0.53/1000 cells, respectively). Thus, the above results indicate that the symptomatic individuals exposed to arsenic through drinking water in this region have significant cytogenetic damage.

Adolescent↗

Fluorescence imaging of reactive oxygen metabolites generated in single macrophage cells (NR8383) upon phagocytosis of natural zeolite (erionite) fibers.

In this paper we address the phenomenon of reactive oxygen metabolite generation subsequent to phagocytosis of mineral fibers by macrophages. Natural erionite fibers were chosen because of their established toxicity. Macrophages (cell line NR8383) were loaded with the dye 5-(and 6)-carboxy-2',7'-dichlorodihydrofluorescein diacetate and exposed to erionite particles by centrifuging cells and fibers together to effect adherence. Reactive oxygen metabolite generation was examined by monitoring the fluorescence of oxidized dye formed via the reaction with oxygen species produced during phagocytosis. Individual cells were repeatedly scanned for up to 2 hr to monitor the evolution of this fluorescence. It was found that erionite-exposed cells had a mean total fluorescence of three times that of controls during the first 35 min, declining to two times that of controls at 35-60 min and about the same level as that of controls at 60-80 min. Ultrastructural studies of similarly treated aliquots of cells showed marked variation in size and numbers of the phagocytized particles. This study demonstrates that intracellular oxidation can be monitored on a single cell basis over a period of time. Quantitative studies are in progress to establish the relationship between the phagocytized particulate load and the extent of fluorescence.

Animals↗

Evaluation of some risk factors for ischaemic heart disease under a matched pairs case-control design.

A pair-wise matched case-control study was carried out in 200 male patients of ischaemic heart disease (IHD). The cases were matched with an equal number of controls in respect of the variables viz., age and socio-economic status. The objective was to quantify the risk of developing IHD for some of the leading risk factors under the univariate and multivariate situations. The analysis revealed that of the factors (smoking, overweight, hypertension, diabetes, serum cholesterol and family history) considered, the smoking habit of the individual, presence of family history of the disease and overweight were the significant risk factors responsible for the development of the disease. The estimated odds ratios were found to be 3.29, 2.60 and 2.00 respectively.

Case-Control Studies↗

In vitro interaction of zeolite fibers with individual cells (macrophages NR8383): measurement of intracellular oxidative burst.

Inhalation of fibrous minerals such as asbestos and erionite can cause various lung diseases, including cancer. The mechanism by which these fibers induce disease is an area of active research. Interaction of fibers with lung macrophages leads to release of many substances. Among these, reactive oxygen metabolites (which include hydrogen peroxide, superoxide, and possibly hydroxyl radicals) are proposed to cause cellular damage. In this paper, we report a method for observing intracellular hydrogen peroxide release as rat lung-derived macrophages (NR-8383) phagocytize erionite fibers. This is possible by observing the fluorescence of 2',7'-dichlorofluorescein-the intracellular, oxidized form of 5 (and 6)-carboxy-2', 7'-dichlorodihydrofluorescin formed in the presence of newly released hydrogen peroxide. We are able to image the fluorescence within a single cell, thereby allowing us to get information on the spatial distribution of the metabolites.

Animals↗

Stereoselective modification of circular dichroism spectra of rat lung beta-adrenoceptor protein preparation by enantiomers of epinephrine.

The presence of beta-adrenoceptor in a rat lung membrane preparation was confirmed by stereoselective competition of enantiomers of epinephrine with labeled iodocyanopindolol. The receptor-rich protein fraction, when combined with the pharmacologically active (-)-epinephrine, exhibited specific changes in the 205-220 nm region of circular dichroism spectra, indicating that the receptor helices may be perturbed. The (+)-epinephrine combined with the lung protein produced little or no change of the spectra. Bovine serum albumin, when combined with either enantiomer of epinephrine, produced nonstereoselective alterations of the spectra. Thus, the data provide important evidence for the higher intrinsic pharmacologic activity of the natural (-)-epinephrine over the unnatural (+)-enantiomer.

Adrenergic beta-Antagonists↗

Profile of tuberculosis--an Army experience.

With the advent of HIV infection an increased awareness has arisen on its effect on the incidence of various diseases. In the Indian context an increased incidence of Tuberculosis has been expected. The Army is a controlled relatively healthy constituent of the general population. It hence provides an ideal sample for gauging any dramatic change in incidence. The present paper covers the secular trend of the disease from the days of independence and highlights the differing rates in the stratum of society within the Army. With increased awareness and improvement in diagnostic facilities an increased incidence of the disease has been noticed. However a prospective study to further study various features of the disease as well its correlation if any with HIV is necessary. The paper also touches upon manpower wastage and age profile of Tuberculosis in the Army, which is an important concern to them.

Adult↗

Analysis of protein denaturation by high-performance continuous differential viscometry.

The intrinsic viscosity of protein solutions changes when denaturation occurs. Therefore, viscosity measurement using traditional glass capillary viscometers has been a key method to study the process of protein denaturation. Such measurements are laborious, time consuming and need at least 10 ml of sample. The Viscotek differential viscometer can be used as an on-line detector for high-performance liquid chromatography to monitor the viscosity of column effluent. In this study, samples were injected using an autosampler onto a "delay" column containing glass beads in place of the high-performance liquid chromatography columns. Results indicate guanidine hydrochloride, heat and pH act as denaturing agents and changes the intrinsic viscosities of serum albumin, turkey egg albumin, and ovalbumin solutions. The differential viscometer is sensitive and provides accurate measurements of minor changes in viscosities of very dilute protein solutions undergoing denaturation. The advantage of using the differential viscometer instead of conventional glass capillary viscometer is the increased sensitivity, precision, speed and operational ease that permits measurements of solution viscosity of low sample concentrations up to 1.2 micrograms of pure proteins.

Animals↗

Characterization of polyamino acids by use of GPC-viscometry technology.

The properties of polybenzyl glutamate, a well known polyamino acid vary considerably with molecular weight in both solid state and in solution. Therefore, accurate determinations of the molecular weights of these polyamino acids is essential. The dual viscometer/refractometer when used as detector system for size exclusion chromatography provides a way of determining accurate molecular weights. An indirect method of determining the molecular weight distribution (MWD) and the radius of gyration distribution (RgD) of polybenzyl glutamate is described. The MWD is calculated from the measured value of intrinsic viscosity (IV) and the known IV-to-MW relationship, at every SEC retention volume slice. Such a technique of determining MWD requires no calibration and is more precisely measurable than conventional SEC methods.

Amino Acids↗

An epidemiological study of malaria in a north-Indian district.

A retrospective epidemiological study was carried out in Gurgaon District of Haryana, wherein data on malaria for a period of more than three decades (1958-1989) obtained from various sources were analysed. The study revealed a rise in the incidence of this disease from midsixties onwards and reached peak in 1976. Subsequently, there has been a gradual and sustained decline, possibly due to better implementation of Modified Plant of Operations (M.P.O.) in a more realistic and rational manner. Various malaria-metric indices like A.P.I., A.B.E.R., S.F.R., etc, have been analysed and attempts made to compare these parameters with that of Haryana State as well as with all India average for last few years. The analysis of the cases according to species, age of patients, seasonal incidence and temporal and spatial distribution have also been discussed.

Adolescent↗

Characterization of temperature-sensitive cytidine triphosphate synthase mutations in bacteria by high-performance liquid chromatography.

Cytidine triphosphate (CTP) synthase catalyzes the last step in pyrimidine ribonucleotide synthesis, namely the formation of CTP from UTP, ATP, and glutamine. Mutants devoid of CTP synthase activity require cytidine for growth and have been designated pyrG in an obligate cdd background. Using a ts mutation blocked in the conversion of UTP to CTP at 43 degrees C, it was demonstrated that the conversion occurs by growing cells at 33 degrees C or below where UTP and CTP pools are normal. Growth at 43 degrees C shuts off the enzyme, while UTP accumulates and CTP is decreased significantly. By now feeding exogenous cytidine the CTP pool can be restored to the level found at the permissive temperature. Intracellular nucleoside triphosphates (CTP and UTP) were separated on a Partisil SAX10 cartridge, using a linear gradient of low buffer (7 mM ammonium dihydrogenphosphate, pH 3.8) to high buffer (250 mM ammonium dihydrogenphosphate, pH 4.5 with 500 mM potassium chloride). Nucleoside triphosphates were also separated after enzymatic conversion of UTP to CTP in solution by cell extracts using ion-pair reversed-phase chromatography on a C18 cartridge eluted with a mixture of 95% buffer A (25 mM ammonium dihydrogenphosphate with 1 mM tetrabutylammonium phosphate, pH 7.0) and 5% buffer B (15% aqueous acetonitrile). Using the two different separation techniques, it was possible to monitor the level of UTP and CTP inside cells as well as the enzymatic conversion of UTP to CTP by the enzyme CTP synthase.

Bacteria↗