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Biomedical subjects

A Haque

Publications and source records attributed to A Haque.

At least 73 records · Page 4Linked to original sources

New findings and symptomatic treatment for neurolathyrism, a motor neuron disease occurring in north west Bangladesh.

Neurolathyrism is a form of spastic paraparesis caused by the neuroexcitatory amino acid 3-N-oxalyl-L-2,3-diaminopropanoic acid (beta-ODAP) present in the seeds and foliage of Lathyrus sativus. The disease is irreversible and usually nonprogressive. Tolperisone HCl, a centrally acting muscle relaxant, has been shown to reduce significantly the spasticity in neurolathyrism patients. Sporadic occurrence of HTLV-1 infection (0.9%) and of osteolathyrism was found among the neurolathyrism patients. Osteolathyrism is linked to the consumption of the green shoots of Lathyrus sativus.

Bangladesh↗

Production of slime polysaccharides by Shigella dysenteriae type 1.

Electron microscopy of ruthenium red-stained ultrathin section of strains of Shigella dysenteriae type 1 grown in the Casamino Acids-yeast extract broth medium showed the presence of an extracellular slime layer. The slime appeared as a dense sheath covering bacteria. The presence of slime promoted hemagglutinating activity of the bacteria. The slime polysaccharide (SPS) isolated from the cell-free culture supernatant or the bacterial surface was less than 162,000 daltons in size and immunochemically similar. The SPS showed cross-reaction with lipopolysaccharide (LPS) antigen in immunological tests; however, it also appeared to be different from LPS since it did not contain 2-keto-3-deoxyoctonate, a core sugar of LPS. A different pattern of separation from LPS was also observed by silver staining of SDS-polyacrylamide gels. From these data it appeared that either LPS and SPS are contaminated with each other or that SPS is the polysaccharide portion of LPS.

Animals↗

Impairment of the cellular immune response in acute murine toxoplasmosis: regulation of interleukin 2 production and macrophage-mediated inhibitory effects.

Depression of the cellular immune response to Toxoplasma gondii has been reported in both mice and humans. The present study was undertaken to determine the kinetics and mechanism of the observed downregulation of interleukin 2 (IL-2) production during experimental murine toxoplasmosis. For these investigations, the cell-mediated immune response to the wild type (PTg) was compared with that to the less-virulent mutant parasite (PTgB), which is deficient in the major surface antigen, p30 (SAG-1). Spleen cells from infected A/J mice failed to proliferate in response to Toxoplasma antigens during the first week of infection. Both PTg- and PTgB-infected A/J mice exhibited a significant reduction in the concanavalin A (Con A)-induced lymphoproliferative response. Further, the response of splenocytes from mice infected with the wild-type parasite was significantly diminished compared with that of mice infected with PTgB. The lymphoproliferative response to Con A reached its nadir at day 7 and remained below control levels for at least 14 days postinfection. By day 21 postinfection, the response to Con A and to Toxoplasma antigens was restored to the level observed prior to day 7. Con A-stimulated culture supernatants of spleen cells from mice on day 7 postinfection contained significantly less IL-2 than normal mice. There was no significant difference in the numbers of binding sites or capacity of high-affinity IL-2 receptors between infected and normal mouse splenocytes as determined by Scatchard analysis. Exogenous IL-2 at different concentrations failed to restore the proliferative response of lymphocytes from infected mice to Con A. Adherent macrophages from 7-day-infected mice were able to suppress IL-2 production by normal splenocytes following stimulation with Con A. The inhibitory activity mediated by infected cells was reversed by the antibody to IL-10 but not transforming growth factor beta. There were insignificant levels of nitric oxide production in both infected and normal splenocytes. These results indicate that during acute murine toxoplasmosis, there is a well-defined period (day 7) during which both the T-cell mitogen and parasite antigen-associated lymphoproliferative response are reduced. Further, there is a reduction in the production of IL-2 and an increase in IL-10, which appear to mediate, in part, the observed downregulation of immunity to T. gondii.

Acute Disease↗

Hemagglutinating properties of enteroaggregative Escherichia coli.

Many intestinal bacterial pathogens possess hemagglutinating properties, which are indicative of their adhesive properties to the intestinal mucosal surface. To understand the bacteria-mucosa interaction, 41 strains of enteroaggregative Escherichia coli (EAggEC), a recently described category of diarrheagenic E. coli, isolated mostly from children with diarrhea in Bangladesh, India, Thailand, Central America, and South America were screened for mannose-sensitive hemagglutination and mannose-resistant hemagglutination of erythrocytes from humans, rats, mice, sheep, cattle, and rabbits. Some strains demonstrated mannose-sensitive hemagglutination of erythrocytes. Most isolates showed mannose-resistant hemagglutination of erythrocytes from all species except rabbits. The hemagglutination patterns could be classified into 18 groups. Studies with three selected isolates suggested that hemagglutinins are cell bound and are protein in nature. On the basis of the pattern of inhibition of hemagglutination by various chemicals, 39 isolates were classified into 19 groups. Hemagglutinations of many isolates were inhibited by sialic acid-containing compounds, suggesting that these compounds may be the receptors for these organisms on erythrocytes and possibly on the intestinal mucosa. These data indicate that strains of EAggEC are a heterogeneous group of organisms with different types of hemagglutinins or adhesins for the intestinal mucosal surface. Also, the adhesion characteristics of EAggEC strains may be too complex to be assessed by simple hemagglutination tests.

Adhesins, Escherichia coli↗

Plasmodium yoelii in mice: antigen reactivity of CD4- and CD8-bearing T cells.

Mice infected with Plasmodium yoelii (265 BY, a nonlethal strain) after recovering from parasitemia become resistant to reinfection. In the present study, we have attempted to define the role of T cell subsets in primary vs secondary P. yoelii infection. We have evaluated the in vivo effects of selective depletion of each subset of T cells on the course of infection and also investigated the in vitro expansion of each subset in response to homologous antigen. Depletion of CD4- or CD8-bearing T cells did not result in reappearance of parasitemia in animals cured from primary infection. However, 25% of reinfected animals treated with anti-CD4 mAb, but not with anti-CD8 mAb, displayed a low level (2 to 3%) of parasitemia late in the secondary infection. The splenocyte response to P. yoelii antigen or to T-cell mitogens was impaired during patient infection, even in the 25% of CD4-depleted animals with low parasitemia. A markedly high lymphocyte reactivity to antigen was observed in mice recovered from primary infection, and this was enhanced in animals exposed to a challenge infection. In the case of animals cured from primary infection, a marked decrease in antigen-induced in vitro lymphocyte proliferation occurred in CD8-depleted but not in CD4-depleted animals when total splenic cell populations were assayed. Although less dramatic, a significant diminution in antigen reactivity was observed also with T-rich populations in CD8-depleted animals. In contrast, there was no such decrease in CD4-depleted mice. Treatment of resistant animals which were challenged with a second infection, with anti-CD4 or anti-CD8 mAbs, resulted in a significant decrease in the proliferative response to antigen by both T-rich and total cell populations. In the secondary infection, the T-rich cell populations from CD8-depleted mice responded better to P. yoelii antigen than the total cell populations, indicating an inhibitory role on the expansion of CD4+ T cells of B cells or their product(s) which were removed during T cell enrichment. The results of our study suggest that CD8-bearing T cells were more reactive in the primary infection. In the secondary infection, although both CD8+ and CD4+ T cells were antigen reactive, the latter T cell subset appeared to play a superior role in controlling the parasitemia.

Animals↗

The antimicrosporidial activity of albendazole.

The antimicrosporidial activity of albendazole was tested on Nosema bombycis in vitro in Spodoptera frugiperda cells and in vivo in Heliocoverpa zea larvae and pupae. Significant reductions in the percentage of infected S. frugiperda cells were obtained using a concentration of 5.3 micrograms/ml albendazole in tissue culture medium but recrudescence occurred after the drug was withdrawn from the cultures. Significant reductions in the number of spores harvested from 6th-instar larvae or pupae were obtained when doses of 0.2 to 4.0 mg were incorporated into the diet but, with the lower doses, some resurgence of infection occurred in pupae after cessation of drug intake. Established infections were almost eliminated from 6th-instar larvae and pupae after consumption of 2 or 4 mg albendazole and infections were not established at all when 4 mg was consumed concurrently with the infective spores. Even at the highest dose albendazole had no deleterious effect on the growth and viability of H. zea. Clumped chromatin in the nuclei of meronts, revealed by electron microscopy, reflected the selective anti-tubulin activity of albendazole and there was massive disorganization of sporogonic development.

Albendazole↗

T-cell recognition of a cross-reactive antigen(s) in erythrocyte stages of Plasmodium falciparum and Plasmodium yoelii: inhibition of parasitemia by this antigen(s).

In the current study, we investigated the presence of a cross-reactive antigen(s) in the erythrocyte stage from Plasmodium yoelii (265 BY strain) and Plasmodium falciparum through recognition by T cells primed in vivo with antigens from each of these parasites. BALB/c mice are naturally resistant to P. falciparum but are susceptible to P. yoelii infection. Mice that had recovered from P. yoelii primary infection became resistant to a second infection. A higher in vitro proliferative response to a soluble blood stage preparation of P. falciparum was observed in splenic cells from immune animals than in those from mice with a patent P. yoelii infection. The antigen-induced proliferative response was enhanced when animals were exposed to a secondary infection. Animals exposed to a challenge infection were treated with anti-CD4 or anti-CD8 monoclonal antibodies to deplete the corresponding subset of T cells. There was a marked diminution in P. falciparum antigen-induced proliferative response in the total splenic cell populations from CD8-depleted but not from CD4-depleted mice. In CD8-depleted and nondepleted animals, the antigen-induced proliferation in the total cell populations was markedly lower than in the T-cell-rich populations, indicating inhibitory activities of B cells and/or macrophages. There was no such difference in the stimulation between total and T-enriched cell populations from CD4-depleted animals. Flow cytometry analysis demonstrated the presence of an almost equal percentage of CD8+ (59.6%) and CD4+ (64%) T cells in the spleen preparations following in vivo depletion of CD4- and CD8-bearing T cells, respectively. When cultured with P. yoelii blood stage antigen, splenocytes from animals immunized with P. falciparum antigen displayed a significant proliferative response which was markedly diminished by treatment with anti-Thy-1.2 antibody plus complement. Animals immunized with P. falciparum antigen and then challenged with P. yoelii blood stage parasites displayed about a 50% lower level of parasitemia. These results demonstrated the existence of a cross-reactive antigen(s) between a murine and a human Plasmodium species, as determined from both in vivo and in vitro biological assays, and indicated the reactivity of mainly CD8+ T cells with this antigen.

Animals↗

Bioregenerative life support as self-sustaining ecosystem in space.

Recent results of research on the development of biological systems and the related technologies for life support ecosystem in space have been reviewed. The integration of higher plants and algal subsystems and the technologies for food and atmospheric regeneration, water reclamation as waste recycling are the aspects which have been dealt with and need critical evaluation before they can be tested in space. The adaptation of living organisms and their ability to multiply and regenerate in space environment during long duration missions is one of the important criteria for their final selection. It is not yet clear to what extent bioregenerative subsystems can be accommodated in the space station Freedom, which will depend on their mass and functional stability, maintainability and operational cost. But it can be expected that bioregenerative life support systems will continue to evolve during the life time of space station Freedom leading towards an integrated ecosystem with further simplification and an increased degree of closure. Several biological and physico-chemical life support systems currently developed are under investigation on the MIR space station.

Animals↗

TNF-alpha production by U937 promonocytes is enhanced by factors released from HIV-infected T4 lymphocytes: TNF-alpha is one of the mediators causing lysis of HIV-infected T4 cells.

In the present study, we have shown that the addition of culture supernatants from HIV-infected SupT1 cells (T4) but not from noninfected cells markedly increased the production of TNF-alpha by U937 promonocytic cells after stimulation with phorbol 12-myristate 13-acetate (PMA). Pretreatment of supernatants with the antibodies to granulocyte/macrophage colony-stimulating factor (GM-CSF) or TNF-alpha, but not interferon-gamma, significantly diminished this enhancing effect. These results suggest that HIV may play an indirect role by producing cytokines from infected T4 cells that can lead to an increased production of TNF-alpha by monocytic cells. Further, TNF-alpha produced by U937 cells following stimulation with PMA plus lipopolysaccharide or with phytohemagglutinin induced lysis of HIV-infected T cells. TNF-alpha-induced cytotoxicity was markedly higher toward HIV-infected than toward noninfected T4 cells. Addition of antibody to TNF-alpha during the cytotoxic phase of response resulted in a reduction of about 50% in the percentage of cytotoxicity, indicating TNF-alpha as one of the lytic mediators.

Antibodies↗

Production, purification, and characterization of botulinolysin, a thiol-activated hemolysin of Clostridium botulinum.

A hemolysin, botulinolysin, produced by Clostridium botulinum was purified to homogeneity and characterized. First, a strain of C. botulinum type C, strain C-203 Tox, which produced a large amount of hemolysin, was selected, and optimal culture medium and conditions for its production of hemolysin were determined. The hemolysin produced in the culture supernatant of this strain under optimal conditions was purified by a combination of ammonium sulfate precipitation, DEAE-Sepharose CL-6B column chromatography, Sephadex G-75 gel permeation chromatography, and SP-Toyopearl 650 M cation-exchange column chromatography, with a recovery of 12%. The purified hemolysin gave a single protein band in polyacrylamide gel electrophoresis (PAGE) with and without sodium dodecyl sulfate (SDS). The protein in this band in PAGE with SDS was estimated to have a molecular weight of 58,000 and was immunostained with a neutralizing monoclonal antibody. In PAGE without SDS, the hemolytic activity corresponded in position to the single protein band. The pI of the hemolysin was 8.4. Amino acid analysis of the purified hemolysin indicated the presence of four half-cystine residues per molecule. The purified hemolysin had a specific activity of 2,100 hemolytic units per microgram of protein on rabbit erythrocytes. It was activated by SH compounds, inhibited by cholesterol, and heat labile. The optimum pH for hemolysis was 6.0 to 7.0. Rabbit, human, and guinea pig erythrocytes were the most susceptible to the hemolysin, while sheep, mouse, rat, and chicken erythrocytes were much less susceptible. The purified hemolysin had a lethal effect in mice and was cytotoxic for some cultured cells: its 50% lethal dose in mice was 310 ng, and its 50% cytotoxic dose for Vero cells was 120 ng/ml.

Amino Acids↗

Independent segregation of glutathione S-transferase and fatty acid ethyl ester synthase from pancreas and other human tissues.

Glutathione S-transferase (GST) isoenzymes of human pancreas were purified, characterized and evaluated for their possible role in the metabolism of ethanol. Human pancreas has at least two GST isoenzymes belonging to the Alpha class (pI 8.8 and 8.1), one belonging to the Mu class (pI 6.4) and one belonging to the Pi class (pI 4.9). During the purification of GSTs from pancreas as well as from heart, liver, lung, brain and muscle, the fatty acid ethyl ester synthase (FAEES) activity was monitored in order to evaluate the role of GSTs in metabolism of ethanol, as suggested in earlier studies. Both t.l.c. and h.p.l.c. were used to identify ethyl oleate in reaction mixtures to monitor FAEES activity. During the purification of GSTs with the use of affinity chromatography on GSH linked to epoxy-activated Sepharose 6B, FAEES and GST activities from each of these tissues segregated independently. Purified GST isoenzymes from these tissues did not exhibit any FAEES activity. Antibodies raised against Pi-class GST, as expected, immunoprecipitated most of the GST activity of brain and heart without precipitating FAEES activity. These results suggest that human GST isoenzymes belonging to the Alpha, Mu and Pi classes do not express FAEES activity. The independent segregation of GST and FAEES activities was further demonstrated by monitoring GST activity during the purification of FAEES from pancreas. It was found that purified FAEES had no GST activity towards 1-chloro-2,4-dinitrobenzene and a number of other electrophilic substrates. Results of these studies demonstrate that FAEES and GSTs are distinct proteins.

Acyltransferases↗

Use of a monoclonal antibody specific for a protein epitope of Echinococcus granulosus antigen 5 in a competitive antibody radioimmunoassay for diagnosis of hydatid disease.

A monoclonal antibody (mAb) designated as EG 02 154/12, specific for the major antigen (antigen 5) of Echinococcus granulosus was produced, and used to study the binding sites recognized by anti-antigen 5 antibodies from patients with hydatid disease. The nature of the target epitope was partially characterized. The antibody reactivity was analyzed towards sheep hydatid fluid antigens (SHF Ag) using ELISA, immunoelectrophoresis (IEP), Western blotting (WB), and immunoprecipitation (IP). In IEP, EG 02 154/12 mAb gave a single precipitin of Ag 5. The mAb and human hydatid patient sera recognized a major antigen of 64 kDa, in SHF Ag analyzed in non-reducing conditions. Both types of antibodies revealed two components of 37 and 22 kDa in reducing conditions. Deglycosylation and delipidation of SHF Ag did not affect the mAb binding. These results, together with the observation of mAb binding to in vitro translation products from protoscoleces messenger RNA, suggest the protein nature of the epitope recognized on the antigen 5. Using competitive antibody radioimmunoassay (CRIA), a competition between this mAb and hydatid patient sera, for the same epitope or closely related sites on antigen 5, was observed. No such competition was detected with the sera from other helminthiasis. The sensitivity and specificity of CRIA was compared to that of ELISA and CRIA found to be an improved diagnostic test for hydatid disease.

Animals↗

A monoclonal antibody to Ly-6 gene product inhibits generation of functionally active T cells and recognizes single antigenic specificity whose expression is up-regulated in virus-transformed rat fibroblast.

In order to elucidate the relationship between the structure and function of proteins encoded for by the Ly-6 gene complex, a cDNA was constructed for a Ly-6.2 specificity and then monoclonal antibodies (mAb) generated to bacterially synthesized protein. The addition of one of these mAb, designated Pb-19, inhibited the proliferative response of T cells to concanavalin A (Con A) or major histocompatability complex (MHC) alloantigens. Reactivity of Pb-19 to the Ly-6 specificity was blocked by a known anti-Ly-6.A.2 mAb but not by an anti-Ly-6.E.1 mAb. This mAb detected a Ly-6.A.2 specificity (a 33,000 MW antigen) whose expression was increased in a transformed rat fibroblast containing the entire genome of bovine papillomavirus.

Animals↗

Use of monoclonal antibodies for the characterization of Onchocerca volvulus antigens.

In order to identify Onchocerca volvulus antigens that could be considered as either diagnostic and/or immunoprophylactic, mouse monoclonal antibodies were produced against O. volvulus soluble antigens. Three were selected on the basis of their staining patterns in an indirect fluorescent antibody assay carried out on cryosections of adult O. volvulus. The first monoclonal antibody (K1-159) recognized a cuticular antigen which appeared in IFA to be restricted to the genus Onchocerca. However, neither Western blotting, nor the immunoprecipitation experiments performed on radiolabelled O. volvulus soluble antigen allowed detection of the corresponding antigen(s). The second monoclonal antibody (K1-126) bound to the muscle cells of adults. A 30,000 Mr antigen was detected by Western blot analysis of adult O. volvulus homogenate. This antigen was also recognised by sera from infected patients and corresponding antigenic determinants were detected in extracts of O. gutturosa, Acanthocheilonema viteae and Ascaris suum, but not in Brugia malayi. The third monoclonal antibody (K1-143) recognized egg shells and the surface of adult worms. The target epitope was not species specific and could be found in O. gutturosa, A. viteae, B. malayi and A. suum. The electrophoretic analysis of I-125 labelled soluble antigens and radiolabelled surface antigens of adult O. volvulus, showed numerous antigens (molecular weights ranging from 30,000 to 120,000 Mr) precipitated by K1-143. In an inhibition radioimmunoassay, K1-143 allowed the detection of corresponding antibodies in 79% of the O. volvulus patient sera tested.

Animals↗

Production of tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) in patients with AIDS. Enhanced level of TNF-alpha is related to a higher cytotoxic activity.

We measured simultaneously circulating and cell-generated TNF-alpha and IL-1 after lipopolysaccharide (LPS) stimulation of peripheral blood mononuclear cells (PBMC) by radioimmunoassay (RIA) in HIV-infected individuals at different stages of infection, classified according to CDC classification. TNF-alpha production, both in vitro and endogenous in sera, remained at the normal level in group II patients but was significantly increased in most patients in group IV (P less than 0.05). Most patients of group II and IV displayed normal level of IL-1 in their sera, whereas the level of this monokine generated in vitro was significantly reduced in both groups (P less than 0.05). The cytotoxic effect of factor(s) secreted by PBMC from HIV-infected individuals was evaluated towards a fibroblast cell line L929. The higher titre of cytotoxicity was directly related to a higher production of TNF-alpha by the cells from group IV patients and the effect could be removed by pre-absorption with anti-TNF-alpha monoclonal antibody.

Acquired Immunodeficiency Syndrome↗

Tolerance in rats by transplacental transfer of Dipetalonema viteae microfilariae: recognition of putative tolerogen(s) by antibodies that inhibit antigen-specific lymphocyte proliferation.

We have previously reported (Nature 1982. 299:361) that the transplacental transfer of Dipetalonema viteae microfilariae (mf) can induce an antigen-specific tolerance in rats. Rats thus tolerized have serum factor(s) which block(s) antigen-specific lymphocyte proliferation. The results of experiments involving fractionation of antisera from tolerant animals indicate that the inhibitory activity for antigen-specific blastogenesis resides in IgG antibodies. Absorption of IgG (eluted from protein A) with specific filarial antigens reduced the inhibition from 58% to 9% whereas a similar immunosorption of IgG size fraction (obtained by applying to AcA 34 Ultrogel) resulted in a decrease from 72% to 35%. This suggests that IgG size fraction might include factor(s) derived from mf and was partially blocking the blastogenic response. Since the tolerant animals harbor only mf, we have used radiolabeled mf surface antigens for immunoprecipitation by antisera from tolerant animals. Antibodies from tolerant animals have a different specificity for filarial antigens compared to those from immunocompetent and mf-resistant rats.

Animals↗

Uptake and accumulation of pentachlorophenol and sodium pentachlorophenate by earthworms from water and soil.

Toxicity values were obtained for pentachlorophenol (PCP) and other compounds in the standard OECD and EEC earthworm test. To help explain the uptake and bioaccumulation mechanisms of PCP which affect toxicity, experiments have been conducted using the earthworm Allolobophora caliginosa. The experiments showed that radiolabelled PCP and its sodium salt, PCP-Na, were taken up rapidly by the earthworms during 24 h from aqueous solution (1 and 10 micrograms ml-1) in a limited volume (5 ml) to give a bioconcentration factor (BCF) of 2.5, and that the solution concentration had no apparent influence on uptake. Exposure of worms over a period of 14 days in a large volume of soil under laboratory conditions increased the BCF, which was approximately 8 and 13 in the low (2.2 mg g-1 soil) and high (11.2 mg g-1 soil) soil concentration respectively. Longer exposures of the same worm species for 131 days in another soil type, in lysimeters under outdoor conditions, did not increase the BCF beyond that reached after 14 days exposure in the artificial soil. There were differences between earthworm species in their bioconcentration ability: Lumbricus terrestris accumulated 3 times more radiocarbon than A. caliginosa. Irrespective of the concentrations in solution, the binding of free phenol (PCP) in worm tissues was greater than for its sodium salt (PCP-Na), but there were no apparent differences in the amount of these substances eliminated, which was rather low. Metabolic studies have shown that PCP is metabolised more rapidly than PCP-Na in the worms body, resulting in the formation of metabolites which are polar in nature and also probably of conjugates. The results are discussed in relation to the mechanisms which may be responsible for the detoxification of PCP in earthworms.

Animals↗