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Biomedical subjects

R Rahman

Publications and source records attributed to R Rahman.

At least 55 records · Page 3Linked to original sources

Detection of Vibrio cholerae O1 in the aquatic environment by fluorescent-monoclonal antibody and culture methods.

Vibrio cholerae O1 in plankton samples collected from ponds and rivers between February 1987 and January 1990 in Matlab, Bangladesh, was detected by the fluorescent-monoclonal antibody (FA) technique. Samples were collected at sites which were monitored fortnightly (fixed sites) as well as at sites that were part of a case-control study. FA results were compared with those obtained by conventional culture methods (CM). A total of 876 samples were collected; V. cholerae O1 was detected in 563 samples (64.27%) by the FA method and in 3 samples (0.34%) by CM. Of the fixed-site plankton samples, 439 (63.62%) were positive by FA and none were positive by CM. Of the 93 case sites sampled on the day after the occurrence of a case of cholera, 73 (78.49%) were positive for V. cholerae O1 by FA and 3 (3.2%) were positive by CM. In comparison, of the 93 first-day sample collections at control sites at the time a case of cholera occurred, only 51 (54.83%) were positive by FA and none were positive by CM. From the data, it is concluded that V. cholerae O1 is present throughout the year in the ponds and rivers of Bangladesh that were examined in this study and that V. cholerae can be detected by FA but not always by CM. The FA procedure was found to be very useful in detecting V. cholerae in plankton, with which it was associated and often occurred in large numbers in the nonculturable stage. Thus, studies investigating the significance of the role of environmental factors in the epidemiology of cholera can be performed effectively by using FA. Such studies are in progress.

Animals↗

Oral cholera vaccines containing B-subunit-killed whole cells and killed whole cells only. II. Field evaluation of cross-protection against other members of the Vibrionaceae family.

Because of demonstrable cross-reactivity of cellular antigens contained in B subunit-killed whole-cell (BS-WC) and killed whole-cell-only (WC) oral cholera vaccines with antigens of various non-cholera species of the family Vibrionaceae (NCV), the protection conferred by the vaccines against diarrhoea associated with NCV was evaluated during a randomized, double-blind field trial in Bangladesh. Children aged 2-15 years and women aged greater than 15 years (62,285 in number) received three doses of BS-WC vaccine, WC-only vaccine, or a placebo consisting of Escherichia coli K12 strain (K12). During 1 year of follow-up, the incidence of treated episodes of diarrhoea associated with non-cholera vibrios known to be enteric pathogens (non-01 Vibrio cholerae, V. fluvialis, V. parahaemolyticus, V. mimicus) in the placebo group was low (1.9 cases per 10,000 recipients) and identical to that for the two vaccine groups combined. The incidence (per 10,000 recipients) of treated diarrhoeal episodes associated with Aeromonas species was considerably higher, but nearly identical in the three groups (26.1 cases for BS-WC, 26.0 cases for WC; 25.9 cases for K12). Pleisiomonas shigelloides was not isolated from any participant. It is concluded that NCV other than Aeromonas were rarely isolated from diarrhoeal patients in our study population and that killed oral vaccines which were effective against cholera exhibited no detectable cross-protection against diarrhoea associated with NCV organisms.

Administration, Oral↗

Monoclonal antibodies to HTLV-III451 gp41: delineation of an immunoreactive conserved epitope in the transmembrane region of divergent isolates of HIV-1.

We report on the development of monoclonal antibodies directed against the transmembrane portion of the envelope of HTLV-III451 gp41. One of these monoclonal antibodies, designated M71/2B4, was found to cross-react with transmembrane proteins from other independent isolates of HIV-1, namely IIIB, MN, and RF. Thus, this monoclonal antibody identifies an epitope located in a region of gp41 that is conserved among all these isolates. To identify this conserved region a series of E. coli recombinant proteins were screened in immunoblot with M71/2B4. From these results the epitope recognized by this antibody appears to map at the amino terminus of gp41, in the region indicated between the cleavage site with gp120 (aa 508) and the HindIII site (aa647).

Animals↗

Immunological and chemical analysis of P6, the carboxyl-terminal fragment of HIV P15.

The first open reading frame of the HIV genome has been identified as the gag gene. The proteins encoded by this gene are p17 as the amino-terminal protein, p24 as the middle peptide, and p15 as the carboxyl-terminal end. A monoclonal antibody recognizing an antigenic determinant on a fragment of p15 has been developed and designated M35/2F8. This monoclonal has been instrumental in radiosequencing the carboxyl-terminal product of p15, p6, and in determining the cleavage site between this protein and the amino-terminal product, p7. By immunoaffinity chromatography it was also possible to purify p6 from HIV lysates and all p6 containing polyproteins from HIV-infected cells. These results gave more insight into the composition and processing of the HIV gag gene.

Antibodies, Monoclonal↗

Characterization of highly immunogenic p66/p51 as the reverse transcriptase of HTLV-III/LAV.

Approximately 80 percent of all human sera that react with antigens of HTLV-III, the etiologic agent of the acquired immune deficiency syndrome (AIDS), recognize protein bands at 66 and 51 kilodaltons. A mouse hybridoma was produced that was specific to these proteins. Repeated cloning of the hybridoma did not separate the two reactivities. The p66/p51 was purified from HTLV-III lysates by immunoaffinity chromatography and subjected to NH2-terminal Edman degradation. Single amino acid residues were obtained in 17 successive degradation cycles. The sequence determined was a perfect translation of the nucleotide sequence of a portion of the HTLV-III pol gene. The purified p66/51 had reverse transcriptase activity and the monoclonal immunoglobulin G specifically removed the enzyme activity from crude viral extract as well as purified enzyme.

Acquired Immunodeficiency Syndrome↗

Monoclonal antibodies specific for p24, the major core protein of human T-cell leukemia virus type III.

Four mouse hybridomas secreting monoclonal antibodies specific for p24, the major core antigen of the human T-cell leukemia virus type III (HTLV-III), have been developed, and their specificities have been partially characterized. These antibodies specifically recognized p24 of HTLV-III in extracts of HTLV-III and in HTLV-III-producing cells. No epitopes cross-reactive with HTLV-I and -II were detected with these antibodies. These hybridomas will be extremely valuable reagents in identifying expression of HTLV-III in infected cultures and in cells or tissues from patients with suspected immunodeficiency syndrome.

Acquired Immunodeficiency Syndrome↗

Distinction of human T-cell line (HUT-102)-derived activity stimulating granulocytic colony formation in diffusion chambers in vivo from activities stimulating erythroid and mixed-colony formation in vitro.

Medium conditioned in the presence of human HUT-102 T-cell line cells contains activities stimulating human mixed (colony-forming unit, erythroid, granulocyte, macrophage, megakaryocyte) and erythroid (burst-forming unit, erythroid) colony formation in methylcellulose in vitro and granulocyte colony formation in diffusion chambers in mice. The stimulatory effect of HUT-102-conditioned medium on colony-forming unit, granulocyte diffusion chamber was also observed in diffusion chambers implanted in nude mice. The hemopoietic activities were heat stable and could be detected from serum-free conditioned medium. Chromatographically, it was possible to separate colony-forming unit, granulocyte diffusion chamber-stimulating activity from activities stimulating burst-forming unit, erythroid and colony-forming unit, erythroid, granulocyte, macrophage, megakaryocyte. On the other hand, the latter two activities were indistinguishable by the methodology used in this study. Failure to abolish the hemopoietic activities by boiling or by human T-lymphotropic retrovirus type 1 antibody indicates that human T-lymphotropic retrovirus type 1 or its components potentially present in the conditioned medium were not responsible for the stimulatory effects.

Cell Line↗

Effects of insulin on CO2 fixation in adipose tissue. Evidence for regulation of pyruvate transport.

Insulin was found to double the rate of incorporation of H14CO3- into protein by segments of rat epididymal adipose tissue provided the incubation medium contained a suitable energy substrate such as fructose. Overall protein synthesis was increased by insulin to a lesser extent, one-third as measured by tritiated water indicating that insulin also increased CO2 fixation into amino acids. The latter could be demonstrated only when the tissue amino acid pools were expanded by the addition of aspartate to the incubation medium. The pattern of labeling observed in the amino acids indicated that CO2 fixation occurred primarily at the pyruvate carboxylase step. Addition of pyruvate to the incubation medium also increased CO2 fixation and this effect was not additive with that of insulin, suggesting that insulin acted by increasing the availability of pyruvate to the carboxylase. No change in carboxylase activity could be measured. Mitochondria isolated from tissue exposed to insulin retained a higher capacity to fix CO2 into acid-soluble products provided they were not freeze-thawed or sonicated. Uptake of pyruvate by mitochondria incubated 1 min at 2 degrees C or 5 s at 15 degrees C was doubled by prior insulin treatment of the tissue. It is concluded that insulin increases the flux through pyruvate carboxylase in adipose tissue in part by increasing the transport of pyruvate through the inner mitochondrial membrane.

Adipose Tissue↗

Ecological relationships between Vibrio cholerae and planktonic crustacean copepods.

Strains of Vibrio cholerae, both O1 and non-O1 serovars, were found to attach to the surfaces of live copepods maintained in natural water samples collected from the Chesapeake Bay and Bangladesh environs. The specificity of attachment of V. cholerae to live copepods was confirmed by scanning electron microscopy, which revealed that the oral region and egg sac were the most heavily colonized areas of the copepods. In addition, survival of V. cholerae in water was extended in the presence of live copepods. Attachment of viable V. cholerae cells to copepods killed by exposure to -60 degrees C was not observed. Furthermore, survival of V. cholerae was not as long in the presence of dead copepods as in the live copepod system. A strain of Vibrio parahaemolyticus was also seen to attach to copepod surfaces without effect on survival of the organism in water. The attachment of vibrios to copepods was concluded to be significant since strains of other bacteria, including Pseudomonas sp. and Escherichia coli, did not adhere to live or dead copepods. Attachment of V. cholerae to live copepods is suggested to be an important factor of the ecology of this species in the aquatic environment, as well as in the epidemiology of cholera, for which V. cholerae serovar O1 is the causative agent.

Animals↗

Male-sterilizing interactions between duplications and deficiencies for proximal X-chromosome material in Drosophila melanogaster.

The genetic limits of sixty-four deficiencies in the vicinity of the euchromatic-heterochromatic junction of the X chromosome were mapped with respect to a number of proximal recessive lethal mutations. They were also tested for male fertility in combination with three Y chromosomes carrying different amounts of proximal X-chromosome-derived material (BSYy+, y+Ymal126 and y+Ymal+). All deficiencies that did not include the locus of bb and a few that did were male-fertile in all male-viable Df(1)/Dp(1;Y) combinations. Nineteen bb deficiencies fell into six different classes by virtue of their male-fertility phenotypes when combined with the duplicated Y chromosomes. The six categories of deficiencies are consistent with a formalism that invokes three factors or regions at the base of the X, one distal and two proximal to bb, which bind a substance critical for precocious inactivation of the X chromosome in the primary spermatocyte. Free duplications carrying these regions or factors compete for the substance in such a way that, in the presence of such duplications, proximally deficient X chromosomes are unable to command sufficient substance for proper control of X-chromosome gene activity preparatory to spermatogenesis. We conclude that there is no single factor at the base of the X that is required for the fertility of males whose genotype is otherwise normal.

Animals↗

Sexual differentiation of hepatic metabolism of corticosterone and testosterone in the rat.

The metabolism of corticosterone and testosterone has been studied in vitro in separate aliquots of the same homogenates of livers from neonatally masculinized and non-masculinized adult castrate male rats. The recovered metabolites were separated by TLC and identified by the combined use of GLC and appropriate derivative formation. The major effect of neonatal masculinization upon the metabolism of both steroids was to increase the ratio of 3 beta : 3 alpha metabolites but this effect was much more prominent in the metabolism of corticosterone than of testosterone. Only 5 alpha metabolites of corticosterone were found, whereas a minor amount of 5 beta products appeared in the case of testosterone. Reduction at C-20 accounted for only about 5% of the metabolism of corticosterone with the exclusive formation of 20 beta-hydroxy derivatives in the non-masculinized liver, but of both 20 alpha and 20 beta products in the masculinized liver. Small amounts of acetylated androstane 3,17-diols and 3-hydroxy-17-ketones were formed, apparently by enzymic reactions that are influenced by neonatal masculinization.

Animals↗

Simple affinity procedure for the purification of mammalian viral reverse transcriptases.

Polyguanylic acid was found to be a potent inhibitor of RNase H associated with mammalian viral reverse transcriptase, indicating a strong interaction between polyguanylic acid and the reverse transcriptase protein. Based on this observation, we have developed three simple procedures for the purification of mammalian viral reverse transcriptases. In the first procedure, a nucleic acid-free extract of Rauscher murine leukemia virus was applied to a column of phosphocellulose and the reverse transcriptase was eluted by a low concentration (50 microM) of polyguanylic acid. Polyadenylic acid and polyuridylic acid could not replace polyguanylic acid for the elution. In the second procedure, a polyuridylic acid-Sepharose column was substituted for phosphocellulose, and the elution was again achieved by polyguanylic acid. In the third affinity procedure, the reverse transcriptase in a nucleic acid-free viral extract was incubated in the cold with 50 microM polyguanylic acid and the complex was adsorbed onto a DEAE-cellulose column. After washing to remove uncomplexed and weakly complexed proteins, the reverse transcriptase was eluted in a concentrated form at 0.3 M NaCl with a recovery of greater than 70%. by polyacrylamide gel analysis in the presence of sodium dodecyl sulfate, the enzyme appeared to be nearly pure.

Chromatography, Affinity↗

Radioimmunoassay for the major structural protein of Mason-Pfizer monkey virus: Attempts to detect the presence of antigen or antibody in humans.

The 25,000 dalton protein of Mason-Pfizer monkey virus (MPMV) was isolated by gel filtration chromatography. In agreement with results from other laboratories, antisera to type-C and the non-type-C bovine leukemia and equine infectious anemia viruses did not precipitate 125I-labelled MPMV p25. In addition, these viruses did not cross-react in a competition radioimmunoassay for MPMV p25. Twenty-one human tissues (15 breast carcinomas, 2 normal breasts, 3 acute myelogenous leukemias and 1 sarcoma) were fractionated by detergent solubilization, ammonium sulfate precipitation, and DE-52 anion exchange chromatography. These methods were shown to be highly effective for purification of MPMV p25. Under assay conditions which minimized incubation damage to the 125I-MPMV p25, all tissues failed to react in the competition radioimmunoassay (RIAT). Two hundred and two human sera or plasma specimens, including those from patients with breast cancer and 33 age-matched controls, from 50 patients with hematologic malignancies, from 12 patients with amyotrophic lateral sclerosis, and from 14 patients with systemic lupus erythematosis, were examined for antibodies to MPMV p25. With the exception of two multiple myeloma plasma which produced artifactual false positive reactions based on hypergammaglobulinemia, a known complication of salt precipitation radioimmunoassays, the remainder of the specimens were negative for evidence of MPMV p25 antibodies.

Adolescent↗

Correlation of human in vivo and in vitro cutaneous antimicrobial factors.

the presence of antimicrobial substances on human skin was investigated. Staphyloccus aureus (10-4 colony-forming units) was applied on the forearm of 50 subjects and covered with a semiocclusive device for 24 hr. In 54% of the subjects the organisms persisted, and in 34% S. aureus was inhibited on the skin. Subjects with persistent S. aureus also had persistent Candida albicans, and vice persa. This correlation was not noted with Streptococcus pyogenes. Skin lipids from the two groups of subjects were extracted with acetone and assayed against S. aureus, S. pyogenes, and C. albicans. The percentage of S. aureus or C. albicans recovered was higher (79% and 55%, respectively) in subjects with persistent microrgamisms on their skin than in those without (47% and 28%, respectively). Subjects with persistent S. aureus and C. albicans had higher counts of normal flora (average, 9.2 times 10-3) than those on whose skin these organisms did not persist (average, 7.4 times 10-2). Coagulase-negative Staphylococcus and Micrococcus were found in higher proportions in subjects with persistent test organisms than in those without. Subjects with lower counts of their normal flora had a higher proportion of diphtheroids (34%) than the high-count group (12%).

Acetone↗

Species and interspecies radioimmunoassays for rat type C virus p30: interviral comparisons and assay of human tumor extracts.

The major internal protein, p30, of rat type C virus (RaLV) was purified and utilized to establish intra- and interspecies radioimmunoassays. Three rat viruses were compared in homologous and heterologous intraspecies assays with no evidence of type specificity. The only heterologous viruses to give inhibition in these species assays were the feline (FeLV) and hamster (HaLV) type C viruses; these reactions were incomplete and required high virus concentrations. An interspecies assay using a goat antiserum prepared after sequentially immunizing with FeLV, RD 114, and woolly monkey virus p30's and labeled RaLV p30 was inhibited by all mammalian type C viruses, although preferentially by RaLV, FeLV, and HaLV. Thus, as in a previously reported assay developed with HaLV p30, rat, hamster, and cat p30's seem more closely related to each other than to mouse type C virus p30. High levels of specific antigen were found in all cell lines producing rat virus, whereas embryonic tissues from several rat strains and cell lines considered virus-free based on other tests were negative for p30. Rats bearing tumors containing Moloney murine sarcoma virus (RaLV) did not contain free circulating antibody to RaLV p30. Fifty-one human tumor extracts (including two tumor cell lines) were tested for activity in the RaLV species and 47 in the interspecies assays after Sephadex gel filtration and pooling of material in the 15,000- to 40,000-molecular-weight range. At a sensitivity level of 7 ng/ml (0.7 ng/assay) in the interspecies assay, all human tissues, with one exception, were negative. The one positive result is considered nonspecific based on proteolysis of the labeled antigen. Input tissue protein of the purified tumor extracts averaged 1.9 mg/ml with a range of less than 0.025 to 22 mg/ml. Tissues from NIH Swiss mice processed in the same manner were positive in the interspecies assay but negative in the intraspecies RaLV assay.

Animals↗