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Biomedical subjects

I Roy

Publications and source records attributed to I Roy.

At least 37 records · Page 2Linked to original sources
PubMed2000

Purification of alpha-amylase isoenzymes from Scytalidium thermophilum on a fluidized bed of alginate beads followed by concanavalin A-agarose column chromatography.

An alpha-amylase has been purified from the thermophilic fungus Scytalidium thermophilum. A ninefold purification was achieved in a single step using fluidized bed chromatography wherein alginate was used as the affinity matrix. There are at least two isoenzymes as shown by concanavalin A (Con A)-agarose column chromatography. The isoenzyme binding to Con A is stable for at least 3 h at 80 degrees C in the presence of calcium ions. The isoenzymes have similar molecular weights of around 45,000 Da as shown by SDS-PAGE analysis. The isoenzymes differ only slightly in their pH optima and temperature optima but the isoenzyme binding to Con A-agarose has slightly higher thermal stability.

Adsorption↗
PubMed2000

Purification of a 'double-headed' inhibitor of alpha-amylase/proteinase K from wheat germ by expanded bed chromatography.

The double-headed inhibitor of alpha-amylase and Proteinase K was purified from wheat germ using Cu(II)-Streamline-chelating resin. The endogenous alpha-amylase could be inactivated by heating. Followed by this step, both packed bed and expanded bed gave similar activity yield of around 83% and fold purification around 23. In the case of expanded bed, it was not necessary to separate precipitated protein before the chromatography. The purified preparation gave a single band on SDS-PAGE and the estimated molecular weight of 21 kDa was in agreement with the reported value for the inhibitor designated as PKI-3 in the literature.

Animals↗
PubMed2000

Clinical and immunologic evaluation of Cedrus deodara pollen: a new allergen from India.

BACKGROUND: Allergy to pollen from gymnosperms is well documented in the West. However, many allergenic species are native to the Himalayan region of India, and Cedrus deodara (Pinaceae) was selected for allergologic investigation. The objective was to define the allergologic and immunochemical aspects of C. deodara pollen. METHODS: Pollen antigen from C. deodara (CD) was prepared and characterized by biochemical and biologic assays. Specific IgE binding was determined by means of ELISA and immunoblotting. RESULTS: CD pollen antigen caused marked skin sensitivity in 7.5% of an atopic population. A significantly elevated level of CD-specific IgE antibodies was observed in 65.8% of the skin-positive patients. Immunoblotting showed protein fractions of 37, 44, 58, and 78 kDa with 100% binding with the patients' sera suspected to be due to carbohydrate moieties. CONCLUSIONS: Patients from the Himalayan region, where CD occurs naturally, were sensitized more than patients from distant places. The immunochemical characterization revealed multiple protein fractions from low to very high molecular mass (14-126 kDa) mostly in the acidic pI range. CD pollen has been recognized as a new allergen from India for the first time. The role of pollen as a causative agent of respiratory allergic disorders is very well established, as is evident from the recent increase of reports from across the world (1-4). India is blessed with the richest flora on the earth, from alpine tundra to Rajasthan desert. Consequently, it provides considerable variation in the quality and quantity of airborne pollen in different ecogeographic regions of the country (5-8). Although studies on the allergenic properties of airborne pollen from various species have been carried out by several workers in India (9-12), information on allergy to aerial pollen from Himalayan tree species has been completely

Allergens↗
PubMed2000

Purification of alkaline phosphatase from chicken intestine by expanded-bed affinity chromatography on dye-linked cellulose.

Alkaline phosphatase from chicken intestine was purified on dye-linked cellulose beads. The various dye-linked cellulose beads were screened in the batch mode for purification of the enzyme and the promising dyes were tried in the expanded-bed mode. The one-step expanded-bed affinity chromatography on Cibacron Blue-linked cellulose beads yielded 48-fold purification with an activity recovery of 70%.

Adsorption↗
PubMed1999

High-level disinfection of gastrointestinal endoscopes: are current guidelines adequate?

OBJECTIVE: For a germicide to obtain a high level disinfection (HLD) claim, FDA requires demonstration of a 6-log reduction of mycobacterial inoculum under worst case conditions. The purpose of this study was to assess the adequacy of current guidelines for high level disinfection of GI endoscopes using alkaline glutaraldehyde in simulated-use testing. METHODS: Various gastrointestinal endoscopes were contaminated with Mycobacterium chelonae in 46 experiments. Quantitative cultures were obtained from each endoscope channel separately after each step: inoculation, standardized manual cleaning, immersion in 2% glutaraldehyde (Cidex) for 10, 20, or 45 min at room temperature, 70% isopropanol rinse, and drying. RESULTS: Manual cleaning alone achieved a 4-log reduction. After 10 min of glutaraldehyde exposure, but before alcohol rinse, two of 10 experiments failed to achieve a 6-log reduction. However, after alcohol rinse, all 10 experiments achieved HLD. After 20 min of glutaraldehyde exposure, but before alcohol rinse, one of 18 experiments failed to achieve a 6-log reduction. After alcohol rinse, all 18 experiments achieved HLD. After 45 min of glutaraldehyde exposure, but before alcohol rinse, one of 18 experiments failed to achieve a 6-log reduction. After alcohol rinse, all 18 experiments achieved HLD. Thus, if the entire reprocessing protocol including manual cleaning, glutaraldehyde exposure, alcohol rinse, and drying was taken into account, the required 6-log reduction of mycobacteria was achieved with a minimum of 10 min of glutaraldehyde exposure at room temperature. CONCLUSIONS: Current guidelines for high level disinfection using glutaraldehyde are appropriate. Alcohol rinse is a valuable adjunctive step for drying and for its bactericidal effects.

Disinfectants↗
PubMed1999

Aerobiology, allergenicity and biochemistry of Madhuca indica Gmel. pollen.

An ASTIR volumetric sampler was used for one year (May 1995-April 1996) for aerobiological survey at Beharampore town, a centrally located representative part of West Bengal, to record the occurrence and frequency of airborne Madhuca pollen. The highest frequency of Madhuca pollen was recorded in April when the weather was dry with low relative humidity (RH) and moderately high temperature. Clinical test (skin prick test) showed Madhuca pollen to be one of the major causes of respiratory allergy. 30-60% (NH(4))(2)SO(4) cut fraction showed maximum positivity in skin prick test. Biochemical analysis showed that Madhuca pollen was rich in lipid and protein. SDS-PAGE was performed with the total soluble pollen protein which showed a total of 6 major protein bands, while in isolated fraction (Fr. II) a total of 7 protein bands were obtained.

Allergens↗
PubMed1998

Aerobiologic and immunochemical studies on Borassus flabellifer pollen: evidence for a 90-kD allergen.

BACKGROUND: The pollen grains of Arecaceae have been proved to be a very common aeroallergen in India. Except for the pollen grains of Cocos nucifera, detailed information about the allergenicity of other dominant Indian palm pollen is not available. OBJECTIVES: To explore the aerobiologic and allergenic significance of the pollen of palmyra palm (Borassus flabellifer Linn.), one of the important and common palm trees in India, with a view to isolate and purify its major allergenic components. METHODS: The aerobiologic survey was performed continuously using Burkard volumetric and Rotorod samplers from July, 1994 to June, 1996 in a northern suburb of greater Calcutta. The allergenic potential of the pollen extract and different fractions were studied by skin prick tests (SPT). ELISA and ELISA inhibition experiments were performed with individual and pooled patient sera to detect the specific IgE level. By 2-step ammonium sulphate fractionation and gel filtration study, the allergic fraction Fr.IIA1 was isolated. Its molecular weight was determined by SDS-PAGE and homogeneity was confirmed by rocket and crossed immunoelectrophoresis using rabbit antisera. RESULTS: Pollen grains of B. flabellifer were found to be present in air from February to May and June contributing more than 7% of the total airborne pollen load of the study area. The pollen elicited maximum concentration at 19 to 20 hours in the evening and at a height of 4 m from ground level. Skin prick testing of 455 respiratory allergic patients with whole pollen extract exhibited 31.64% positive response. After 2-step ammonium sulphate fraction and gel filtration, a fraction (Fr.IIA1) having remarkable allergenicity was isolated. In native and SDS-PAGE, it was found as a single protein component of 90 kD and as homogeneous in RIE and CIE. The soluble protein-carbohydrate analysis indicated the probability of the component to be a glycoprotein. CONCLUSION: The aerobiologic, clinical, and immunochemical, studies demonstrated that the pollen of B. flabellifer is a dominant aeroallergen to cause respiratory trouble in the area where these trees grow. The isolated 90-kD component is one of the major allergens present in the pollen extract.

Adult↗
PubMed1998

Phosphorylation of the periplasmic binding protein in two transport systems for arginine incorporation in Escherichia coli K-12 is unrelated to the function of the transport system.

In Escherichia coli K-12, the accumulation of arginine is mediated by two distinct periplasmic binding protein-dependent transport systems, one common to arginine and ornithine (AO system) and one for lysine, arginine, and ornithine (LAO system). Each of these systems includes a specific periplasmic binding protein, the AO-binding protein for the AO system and the LAO-binding protein for the LAO system. The two systems include a common inner membrane transport protein which is able to hydrolyze ATP and also phosphorylate the two periplasmic binding proteins. Previously, a mutant resistant to the toxic effects of canavanine, with low levels of transport activities and reduced levels of phosphorylation of the two periplasmic binding proteins, was isolated and characterized (R. T. F. Celis, J. Biol. Chem. 265:1787-1793, 1990). The gene encoding the transport ATPase enzyme (argK) has been cloned and sequenced. The gene possesses an open reading frame with the capacity to encode 268 amino acids (mass of 29.370 Da). The amino acid sequence of the protein includes two short sequence motifs which constitute a well-defined nucleotide-binding fold (Walker sequences A and B) present in the ATP-binding subunits of many transporters. We report here the isolation of canavanine-sensitive derivatives of the previously characterized mutant. We describe the properties of these suppressor mutations in which the transport of arginine, ornithine, and lysine has been restored. In these mutants, the phosphorylation of the AO- and LAO-binding proteins remains at a low level. This information indicates that whereas hydrolysis of ATP by the transport ATPase is an obligatory requirement for the accumulation of these amino acids in E. coli K-12, the phosphorylation of the periplasmic binding protein is not related to the function of the transport system.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1998

Carbodi-imide coupling of enzymes to the reversibly soluble insoluble polymer Eudragit S-100.

The coupling of proteins and enzymes to soluble-insoluble polymers by carbodi-imide can be performed by using numerous variations of the protocol. This protocol has been investigated for the coupling of five different enzymes, namely wheatgerm acid phosphatase, beta-glucosidase, beta-galactosidase, trypsin and xylanase, to an enteric methacrylate polymer Eudragit S-100. The following results were found. (1) The activity of the bioconjugate was critically dependent on the physical state of the polymer and the pH of the coupling reaction. For example, in the case of wheatgerm acid phosphatase, the activity of the bioconjugate was 49% when coupling was performed at pH 7.2 and 67% when coupling was performed at pH 4. 5. With beta-galactosidase the corresponding values were 57% and 23% and with beta-glucosidase they were 57% and 52% respectively. (2) In some cases, such as beta-glucosidase and beta-galactosidase, it might be necessary to remove excess carbodi-imide before the addition of the enzyme to the activated matrix. (3) In most of the cases investigated, a sig-nificant amount of the enzyme (more than 90%) could be bound to the matrix merely by adsorption. (4) More importantly, after the carbodi-imide coupling procedure, a sufficient fraction of the bound enzyme could be eluted off the matrix, indicating that this was merely adsorbed and not covalently coupled.

Acid Phosphatase↗
PubMed1997

Necrobiotic palisading suture granulomas involving bone and joint: report of two cases.

Biomaterial used in surgery is relatively inert and non-toxic; however, adverse reactions may follow implantation of such foreign material. We describe the first two cases of bone and joint destruction by necrobiotic palisading suture granulomas. The hypersensitivity reaction occurred years after shoulder repair using silk sutures. One patient received chemotherapy for a mistaken diagnosis of tuberculous arthritis. Although very rare, foreign material should be included in the differential diagnosis of necrotizing granulomas. A history of surgery and microscopic examination with polarized light should allow recognition of this entity.

Adult↗
PubMed1996

The masked cysteine residues in methylmalonyl-CoA mutase from Propionibacterium shermanii are essential for catalytic activity.

Two masked cysteine residues have been reported in methylmalonyl-CoA mutase from Propionibacterium shermanii, Cys-535 in the alpha-subunit and Cys-517 in the beta-subunit, which are revealed only after reduction of the denatured enzyme with dithiothreitol. It has been postulated that these residues are involved in disulphide linkages to unknown thiols of low M(r). These two masked cysteine residues have been changed to an alanine, individually. Both the mutants, C535alphaA and C517betaA, were inactive. This shows that both these residues are essential for catalytic activity.

Alanine↗
PubMed1993

Human studies to measure the effect of antibiotic residues.

This epidemiological study compares the frequency of resistant bacteria in stool microflora among vegetarians and nonvegetarians over a 12 month period. Two well characterized vegetarian populations (one in Boston, MA and the other in Loma Linda, CA) as well as appropriate controls were studied. No apparent differences in the prevalence of antibiotic resistance in the microflora were noted; however, vegetarians had a significantly greater incidence of multi-antibiotic resistance. E. coli of the same API biotype had the same frequency of antibiotic resistance in both vegetarians and nonvegetarians. Quantitative studies showed similar percents of tetracycline resistant facultative isolates and of "bacteroides." Klebsiella were more common in the stool of the nonvegetarians. As shown in previous studies, exposure to animal products either as meat eaters or production workers in a poultry abattoir was not associated with an increased incidence of resistant bacterial flora or infections caused by resistant strains.

Anti-Bacterial Agents↗
PubMed1990

Diversity and stability of restriction enzyme profiles of plasmid DNA from methicillin-resistant Staphylococcus aureus.

Nosocomial infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are a significant epidemiological problem. Detecting the sources of epidemic strains and preventing their access to patients, however, depend upon the availability of techniques to reliably distinguish among MRSA strains. We evaluated restriction enzyme analysis of plasmid DNA for use as an epidemiological marker of MRSA strains. The diversity of plasmid types was assessed by examining 120 clinical and environmental MRSA isolates from five southern California hospitals and from the American Type Culture Collection. Thirty-seven distinctive EcoRI digestion patterns were observed. We characterized each strain by the number of plasmids it contained and the sizes of the fragments that were generated by EcoRI. Very few of the isolates (4.2%) lacked plasmids, and some (6.7%) contained DNA that was not digested by EcoRI. Several isolates (12.5%) contained two or more plasmids. We were able to assess the stability of MRSA plasmid types by tracking epidemic strains over a 2-year period. We also examined successive isolates from 10 individual patients during their hospitalization. In all but one case, the patient's plasmid profiles remained unchanged. We conclude that the diversity and stability of MRSA plasmid types make them excellent epidemiological markers. In support of this conclusion, we found that our data provided significant epidemiological insights. Two epidemic strains, accounting for more than half of the infections, were identified in the five hospitals. The remaining cases were sporadic, caused by MRSA strains that appeared very infrequently and that may have originated from sources outside the hospitals.

Cross Infection↗
PubMed1990

Hispanic Health and Nutrition Examination Survey: methodological considerations.

The Hispanic Health and Nutrition Examination Survey (HHANES) was the first special population survey undertaken by the National Center for Health Statistics. The HHANES was designed to assess the health and nutritional status and needs of Mexican Americans, mainland Puerto Ricans and Cuban Americans. Data were collected using five data collection techniques: direct physical examinations, diagnostic testing, anthropometry, laboratory analyses, and interviews. Unlike other surveys conducted by the National Center for Health Statistics, the HHANES was not designed as a national survey. The HHANES was a survey of three Hispanic subgroups of the population in selected areas of the United States with a survey universe that included approximately 76 percent of the 1980 Hispanic-origin population in the United States. This article discusses statistical issues that should be addressed by researchers when analyzing HHANES data. Specifically, analysts need to account for the complex sample design, nonresponse bias, potential non-coverage bias, and the regional nature of the HHANES sample.

Acculturation↗