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H Ahmad

Publications and source records attributed to H Ahmad.

At least 73 records · Page 4Linked to original sources

Anionic glutathione S-transferases of human erythrocytes, placenta, and lung: evidence for structural differences.

Studies were undertaken to elucidate the structural interrelationships among glutathione S-transferase (GST) isozymes of human placenta, lung, and erythrocytes. Results of the high-performance liquid chromatography of the trypsin digests of the three isozymes indicate minor but significant differences in their elution profiles. Although a number of peptides generated by proteolysis were common for either 2 or 3 of the isozymes, significant differences were observed in elution profiles of other peptides. Qualitative as well as quantitative differences were also observed in the electrophoretic peptide maps of these isozymes. These studies suggest that there may be fine structural differences among the pi class GST isozymes of human tissues.

Amino Acid Sequence↗

Glutathione S-transferase of bovine iris and ciliary body: characterization of isoenzymes.

Five forms of glutathione (GSH) S-transferase (GST) having catalytic activities towards a variety of xenobiotics were present in bovine iris-ciliary body. In contrast to that in lens, cornea, and retina, GST isoenzymes belonging to all the three classes (alpha, mu and pi) were present in iris as well as in the ciliary body. GST isoenzymes of iris-ciliary body had pI values of 8.7, 7.4, 7.0, 6.6, and 6.0. GST 8.7 and GST 7.4 were apparent homodimers of 27,000 and 22,500 Mr subunits, respectively. GST 8.7 cross-reacted only with antibodies raised against the alpha class GST of human liver and GST 7.4 cross-reacted with the antibodies raised against GST pi of human placenta. GST 7.0 and 6.6 were heterodimers of Mr 26,500 and 25,000 subunits and both these subunits cross-reacted with the antibodies raised against the mu class human GST. Iris-ciliary body contained both, GSH peroxidase I and GSH peroxidase II activities and in this respect also, they differ from lens, cornea, and retina each of which have only one of these two activities. The presence of several GST isoenzymes belonging to all the three major classes and both GSH peroxidase I and II activities in iris-ciliary body may be important for the detoxification of oxidants and xenobiotics in order to prevent their infiltration in aqueous humor.

Animals↗

Omeprazole compared with ranitidine once daily in the treatment of duodenal ulcer.

A double-blind randomized study in 230 Malaysian patients with duodenal ulcer was conducted to compare the proton-pump inhibitor, omeprazole 20 mg, given once daily in the morning, with ranitidine 300 mg, administered once daily at night. After 2 and 4 weeks of treatment, 222 and 220 patients, respectively, were evaluable according to the study protocol. Omeprazole produced significantly higher healing rates than ranitidine at both 2 weeks (75% versus 46%, respectively, P less than 0.0001) and 4 weeks (97% versus 83%, respectively, P = 0.001). Ulcer symptoms were relieved more rapidly by omeprazole than ranitidine. After 2 weeks, daytime epigastric pain was reported by 30% of ranitidine-treated patients but only by 15% of omeprazole-treated patients, which is a statistically significant difference (P = 0.004). No major clinical or biochemical side effects were recorded for either omeprazole or ranitidine. In conclusion, omeprazole 20 mg was found to be superior to ranitidine 300 mg administered once daily for the treatment of duodenal ulcer as measured by ulcer healing and pain relief.

Adult↗

Differential expression of alpha, mu and pi classes of isozymes of glutathione S-transferase in bovine lens, cornea, and retina.

Isozyme characterization of glutathione S-transferase (GST) isolated from bovine ocular tissue was undertaken. Two isozymes of lens, GST 7.4 and GST 5.6, were isolated and found to be homodimers of a Mr 23,500 subunit. Amino acid sequence analysis of a 20-residue region of the amino terminus was identical for both isozymes and was identical to GST psi and GST mu of human liver. Antibodies raised against GST psi cross-reacted with both lens isozymes. Although lens GST 5.6 and GST 7.4 demonstrated chemical and immunological relatedness, they were distinctly different as evidenced by their pI and comparative peptide fingerprint. A corneal isozyme, GST 7.2, was also isolated and established to be a homodimer of Mr 24,500 subunits. Sequence analysis of the amino-terminal region indicated it to be about 67% identical with the GST pi isozyme of human placenta. Antibodies raised against GST pi cross-reacted with cornea GST 7.2. Another corneal isozyme, GST 8.7, was found to be homodimer of Mr 27,000 subunits. Sequence analysis revealed it to have a blocked amino-terminus. GST 8.7 immunologically cross-reacted with the antibodies raised against cationic isozymes of human liver indicating it to be of the alpha class. Two isozymes of retina, GST 6.8 and GST 6.3, were isolated and identified to be heterodimers of subunits of Mr 23,500 and 24,500. Amino-terminal sequence analysis gave identical results for both retina GST 6.8 and GST 6.3. The sequence analysis of the Mr 23,500 subunit was identical to that obtained for lens GSTs. Similarly, sequence analysis of the Mr 24,500 subunit was identical to that obtained for the cornea GST 7.2 isozyme. Both the retina isozymes cross-reacted with antibodies raised against human GST psi as well as GST pi. The results of these studies indicated that all three major classes of GST isozymes were expressed in bovine eye but the GST genes were differentially expressed in lens, cornea, and retina. In lens only the mu class of GST was expressed, whereas cornea expressed alpha and pi classes and retina expressed mu and pi classes of GST isozymes.

Amino Acid Sequence↗

A novel dinitrophenylglutathione-stimulated ATPase is present in human erythrocyte membranes.

Vesicles prepared from human erythrocyte membranes were found to catalyze ATP hydrolysis that was stimulated by dinitrophenylglutathione (Dnp-SG). This activity was dependent on temperature and Mg2+ and independent of ion pump ATPases present in erythrocyte membranes. The activity was a linear function of protein and time up to 60 min. The Km values of ATPase for Dnp-SG and ATP were found to be 49 microM and 1.67 mM, respectively. This suggests that in erythrocytes, the transport of Dnp-SG requires direct enzymatic hydrolysis of ATP and both Dnp-SG-stimulated ATPase activity and the ATP-dependent efflux of Dnp-SG from erythrocytes represent different activities of the same protein.

Adenosine Triphosphatases↗

Purification and characterization of unique glutathione S-transferases from human muscle.

Results of studies designed to investigate the origin of the diversity of glutathione S-transferase (GST) isozymes in human tissues indicated that human muscle has at least three forms of GST with pI values of 5.0, 5.1, and 5.2 that are distinct from GST isozymes characterized so far. The major muscle isozyme which was expressed in all the six samples analyzed in this study was a unique GST of pI 5.2 that was designated as GST zeta. It had a blocked N-terminal and did not correspond to any of the known three classes (alpha, mu, or pi) of human GST as evidenced by its immunological properties and substrate specificities. The N-terminal regions of human muscle GST 5.1 and 5.0 had identical amino acid sequences except at residue 5, but demonstrated significant differences in amino acid composition and substrate specificities. These two isozymes showed homology with the mu class of human GST in their N-terminal region and were also immunologically related to the mu class of human GST although their subunit molecular weight values (Mr 23,000) were lower than that reported for GST psi. The substrate specificities of these isozymes were also significantly different from those of other human GST isozymes characterized so far. Significantly, muscle tissue did not express the alpha class of GST isozymes; however, two other isozymes were identified, GST 4.8 and GST 4.5, which had identical N-terminal amino acid sequences that were similar to that reported for the pi class of human GST. GST 4.8 was present in all six samples analyzed in this study whereas GST 4.5 was present in only two of these samples, indicating a possibility of polymorphism at the GST3 locus. This study indicated the occurrence of at least three distinct isozymes in muscle tissue, providing further evidence for tissue specific expression of GST isozymes in humans.

Amino Acid Sequence↗

Expression of glutathione S-transferase isoenzymes in human small cell lung cancer cell lines.

A glutathione S-transferase (GST) isoenzyme having common antigenicity to rat placental form (GST-P) and human placental form (GST-pi) has recently been suggested may be a marker of carcinogenesis. In the present study we have investigated the expression of this isoenzyme in three small cell lung cancer cell lines in order to determine whether or not this isoenzyme can be used as a general marker of carcinogenesis. GST activity towards 1-chloro-2,4-dinitrobenzene in two of the cell lines (NES and NOC-361) was moderately higher than that in normal human lung, but this activity was markedly suppressed in one of the cell lines (NCI-H69). Quantitation of the GST isoenzymes in the tumors grown in nude mice by injecting these cell lines also revealed a moderate increase of GST-pi-type isoenzyme in NES and NOC-361 and its suppression in NCI-H69. Immunocytochemical localization studies with these tumors using antibodies raised against GST-pi also indicated a drastic decrease of GST-pi-type isoenzyme in NCI-H69 and this finding was confirmed by Western blot studies. These results suggest that GST-pi, or the isoenzyme(s) having similar immunological nature to GST-pi, cannot be used as the general markers of neoplastic states.

Animals↗

Glutathione S-transferase isoenzymes in human lung tumors.

In the present studies we have compared the levels of glutathione (GSH) and GSH-related enzymes in lung tumors and corresponding normal tissues obtained from the same individuals. We have also immunologically quantitated the relative amounts of glutathione S-transferase pi (or GST-P) type antigen in tumors and adjacent normal tissues from five patients. GST activities towards 1-chloro-2,4-dinitrobenzene (CDNB) and ethacrynic acid were found to be elevated in tumors from two out of five patients (patients #1 and 4), whereas the activity towards these substrates was markedly suppressed in the tumor tissue from one of the patients (#5). Immunotitration and Western blot studies using antibodies raised against pi-type GST isoenzymes of human lung and placenta indicated induction of GST pi-type isoenzyme in tumors from patients #1 and 4 and suppression of this isoenzyme in tumor from patient #5. The tumors from patients #2 and 3 did not show any increase in GST activity or GST pi-type antigen. Except for the tumor from patient #5, the GSH content was higher in the tumors from other patients. GSH reductase activity was found to be elevated in tumors of all the patients examined in this study. These results indicate that GSH and GSH related enzymes are differentially altered in lung tumors and GSH levels and GST pi- or GST-P-type isoenzyme(s) are not uniformly elevated in all tumors.

Carcinoma↗

Immunocytochemical evidence for the expression of GST1, GST2, and GST3 gene loci for glutathione S-transferase in human lung.

Immunocytochemical studies demonstrate that significant amounts of glutathione S-transferase (GST) are associated with alveoli and bronchioles of human lung. The immunofluorescence in human lung sections was observed with the antibodies which were raised against GST psi and GST alpha-epsilon of human liver and GST pi of human placenta indicating that the isoenzymes corresponding to three gene loci, GST1, GST2, and GST3 are present in human lung. Presence of GST isoenzymes in significant amounts in bronchioles and alveoli of human lung indicate that these isoenzymes may play an important role in the detoxification of xenobiotics as well as in combating oxidative stress through glutathione peroxidase II activity.

Bronchi↗

A Coomassie blue-binding assay for the microquantitation of immobilized proteins.

A sensitive assay procedure for the determination of microgram quantities of immobilized proteins is described. The procedure is based on the property of Coomassie blue G-250 to bind strongly yet reversibly to proteins. The assay involves incubation of the immobilized protein with a solution containing 0.1% Coomassie blue, 10% acetic acid, and 25% isopropyl alcohol in distilled water at room temperature followed by washing off of the unbound dye. The protein-bound dye is eluted with methanolic NaOH, acidified, and the absorbance is measured at 605 nm. The assay is highly reproducible and several proteins immobilized on various matrices could be conveniently assayed. Protein values determined by the dye-binding assay showed good agreement with those obtained by other procedures.

Amines↗

Bromophenol blue protein assay: improvement in buffer tolerance and adaptation for the measurement of proteolytic activity.

A modification of the bromophenol blue dye binding procedure of protein estimation is described. Substitution of glycine/phosphoric acid, pH 2.6, for dilute acetic acid in the colour reagent extended the applicability of the procedure to protein solutions containing buffers of various pH values. This was, however, accompanied by approximately 25% loss in the sensitivity of the procedure. The modified reagent exhibited very marked tolerance to detergents and could be successfully adapted for the measurement of proteolytic activity in acidic, neutral or alkaline pH ranges.

Bromphenol Blue↗

Studies on the migratory route of Paragonimus siamensis in the bandicoot (Bandicota indica).

The migration route and the development of P. siamensis obtained from S. juliae and S. germaini were studied in B. indica. The infection rates of P. siamensis in these 2 species of crabs were 19.7% and 21.4% respectively. Worms were recovered in the abdominal cavity, pleural cavity and lungs on day 12, 16 and 30 respectively after oral infection. Worms became mature and laid eggs on day 47 after infection, but their size still increased considerably. The average egg output was found to be 144 eggs/gm faeces/worm or 443 eggs/day/worm. The average recovery rate of P. siamensis in the experimental B. indica was 15%.

Animals↗

ATP dependent primary active transport of xenobiotic-glutathione conjugates by human erythrocyte membrane.

We have demonstrated the presence of a dinitrophenyl glutathione (Dnp-SG) stimulated ATPase in human erythrocyte membranes. This ATPase mediates the active transport of glutathione-xenobiotic conjugate such as Dnp-SG from erythrocytes into the plasma. It is suggested that this transport system is distinct from the system which actively transports oxidized glutathione from the erythrocytes.

Adenosine Triphosphatases↗

Epidemic dengue transmission in southern Sumatra, Indonesia.

An outbreak of dengue fever (DF), dengue haemorrhagic fever (DHF), and dengue shock syndrome (DSS) in the city of Palembang, south Sumatra, Indonesia was investigated to (i) validate epidemic occurrence, (ii) confirm dengue virus aetiology and associated serotype(s), (iii) provide a demonstrable measure of community impact, and (iv) identify causative relationship (if any) with climatic El Niño Southern Oscillation (ENSO) influences. Trend analysis based on a 6-year retrospective review of hospital records demonstrates a 3-fold increase in clinical cases for the outbreak period (January-April 1998), relative to historical records. In the 2 hospitals surveyed, the monthly mean number of outbreak-related dengue cases over 4 months was 833 (range 650-995 cases/month); the mean monthly value for the previous 72 months was 107 (range 14-779 cases/month). An apparent trend in epidemic transmission was observed, evolving from a 5-year cyclic phenomenon to an annual occurrence, often indistinguishable from one year to the next. The proportional distribution of clinical outbreak cases into DF, DHF and DSS diagnostic categories was 24%, 66%, and 10%, respectively. The population aged 10-19 years accounted for the largest (35%) proportion of hospitalized DHF cases, followed by children aged 5-9 years (25%) and children aged 4 years (16%). Serum samples obtained during acute illness from 221 hospitalized patients were examined using serology, RT-PCR, and virus isolation in cell culture: 59% of samples had laboratory evidence of a dengue infection. All 4 dengue virus serotypes (DEN 1-4) were identified in epidemic circulation, with DEN 3 predominating (43%). DEN 1 was the principal serotype associated with less severe dengue illness, suggesting that virulence may be, in part, a function of infecting serotype. The climatic influence of ENSO on rainfall and temperature in the months leading up to and during the outbreak was dramatic, and is likely to contribute to favourable outbreak conditions.

Adolescent↗

In vitro sensitivity of medically significant Fusarium species to various antimycotics.

Sixteen isolates belonging to Fusarium chlamydosporum (n = 4), Fusarium equiseti (n = 1), Fusarium moniliforme (n = 2), Fusarium oxysporum (n = 3), Fusarium proliferatum (n = 1), and Fusarium solani (n = 5) were tested against amphotericin B, 5-fluorocytosine, fluconazole, itraconazole, ketoconazole, JAI-amphotericin B (water-soluble compound), hamycin and amphotericin B combined with 5-fluorocytosine, using antibiotic medium M3, high-resolution broth (pH 7.1), Sabouraud's dextrose, and yeast-nitrogen broth media (1 ml/tube). The minimal inhibitory and minimal fungicidal concentrations of 5-fluorocytosine and fluconazole for all species were > 100 micrograms/ml. All Fusarium isolates, except F. equiseti (3.125 micrograms), gave minimal inhibitory concentrations of 12.5-100 micrograms/ml for hamycin. The values for amphotericin B, itraconazole, ketoconazole, JAI-amphotericin B, and amphotericin B combined with 5-fluorocytosine were 1.56-100, 0.78-50, 3.125-100,50-100, and 1.56 to > 100 micrograms/ml, respectively. Although a wide range of minimal inhibitory concentrations was recorded for most of the isolates studied, it appears that some--F. solani, F. oxysporum, F. chlamydosporum, F. equiseti, and F. moliniforme--were more susceptible to amphotericin B, itraconazole, ketoconazole, hamycin, and amphotericin B in the presence of 5-fluorocytosine. All isolates showed resistance to 5-fluorocytosine and fluconazole. The minimal fungicidal concentrations were either the same or several times higher than the minimal inhibitory concentrations.

Antifungal Agents↗