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Biomedical subjects

I A Khan

Publications and source records attributed to I A Khan.

At least 19 recordsLinked to original sources
PubMed1992

Antigen-specific (p30) mouse CD8+ T cells are cytotoxic against Toxoplasma gondii-infected peritoneal macrophages.

The importance of CD8+ T cells in immunity against Toxoplasma gondii is now well recognized. The mechanism by which these CD8+ T cells are able to confer this immunity is not yet understood. To examine the Ag specificity of this response, immune splenocytes from mice immunized with p30, a major surface parasite Ag, were evaluated for their ability to lyse peritoneal macrophages infected with three different strains of T. gondii. Macrophages infected with either the RH or P wild-type strain tachyzoites were lysed at varying E:T ratios by nylon wool nonadherent immune splenocytes whereas macrophages infected with a p30-deficient mutant (B mutant) of the P strain were not. The gene encoding p30 for the wild type and B mutant were amplified by the polymerase chain reaction. This revealed a nonsense mutation in the B mutant such that its primary translation product is predicted to be about two-thirds the size of the wild-type p30 molecule. mAb depletion studies indicate that the cytotoxic effect of the immune splenocytes is mediated by the CD8+ T cell population. Peritoneal macrophages infected with the three different strains (RH, P wild type, B mutant) from mice genetically restricted were not lysed by the immune CD8+ effector cell population. A cloned line (C3) of p30 Ag-specific CD8+ T cells exhibited significant cytotoxicity against syngeneic peritoneal macrophages infected with either the RH or P strain tachyzoites. There was no macrophage lysis observed by these CD8+ effector cells of either syngeneic macrophages infected with the B mutant or nonsyngeneic macrophages infected with the three different tachyzoite strains.

Amino Acid Sequence↗
PubMed1992

Stimulatory effects of serotonin on maturational gonadotropin release in the Atlantic croaker, Micropogonias undulatus.

The effects of serotonin (5-HT) injection alone, and in combination with des Gly10 [D-Trp6]-luteinizing hormone-releasing hormone ethylamide (LHRHa), on plasma maturational gonadotropin (GtH) levels in the Atlantic croaker, Micropogonias undulatus, were examined. An injection of 5-HT (20 micrograms/g body wt) alone did not significantly elevate GtH levels, whereas simulatneous administration of LHRHa (20 ng/g body wt) and 5-HT elicited a greater elevation of plasma GtH levels than that induced by LHRHa alone. Pretreatment of 1-year-old fish with fluoxetine (10 micrograms/g), a 5-HT reuptake inhibitor, only slightly augmented the effect of 5-HT on GtH levels, whereas pretreatment with ketanserin (10 micrograms/g), a 5-HT receptor antagonist, completely inhibited the potentiating effect of 5-HT on the GtH response to LHRHa. Administration of LHRHa (20 ng/g) or 5-HT (20 micrograms/g) significantly elevated GtH levels in the 2- and 3-year-old croaker, but the combined treatment failed to increase GtH levels above those induced by LHRHa alone. However, with a lower dose of LHRHa (5 ng/g), the combination produced an additive effect. Serotonin (20 micrograms/ml media) alone, and in combination with LHRHa (10 ng/ml media), significantly stimulated GtH release from the pituitaries of gonadally mature 2- and 3-year-old female croaker during an 18-hr incubation in vitro, but not from pituitaries of gonadally regressed fish. The combined treatment of LHRHa with 5-HT also significantly stimulated in vitro GtH release during 6- and 12-hr pituitary incubations, whereas these treatments alone were ineffective.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1992

Tubulin structure and biochemistry.

In the past year, much has been learned about structure-function correlations in the tubulin molecule, and specifically about the nature and roles of post-translational modifications and tubulin isotypes. The interactions between tubulin and its ligands--both microtubule-associated proteins and anti-mitotic drugs--are becoming clearer at the molecular level.

Animals↗
PubMed1992

New phenolic glucosides from the leaves of Eurya tigang.

Three new compounds, 6'-O-coumaroyl-1'-O-[2-(4-hydroxyphenyl)ethyl]-beta-D-glucopyra nos ide [1] (eutigoside A), 6'-O-coumaroyl-1'-O-[2-(1-hydroxy-4-oxo-2,5-cyclohexadien-1- yl)ethyl]-beta-D-glucopyranoside [2] (eutigoside B), and 6'-O-cinnamoyl-1'-O-[2-(1-hydroxy-4-oxo-2,5-cyclohexadien-1-yl)eth yl]- beta-D-glucopyranoside [3] (eutigoside C) have been isolated from the leaves of Eurya tigang, along with other known compounds (afzelin, quercitrin, p-coumaric acid, methyl-alpha-D-fructofuranoside, isorengyol, and euryanoside). Their structures were determined by chemical and spectroscopic methods (uv, ir, ms, 1H-1H COSY, and 1H-13C COSY).

Animals↗
PubMed1991

A purified parasite antigen (p30) mediates CD8+ T cell immunity against fatal Toxoplasma gondii infection in mice.

Induction of protective immunity against acute and chronic toxoplasmosis can be achieved using p30, the major membrane and excreted/secreted protein of Toxoplasma gondii. This protein, when administered to outbred mice in the presence of the saponin Quil A, is able to induce almost 100% protection against acute infection without evidence of intracerebral cyst development. Adoptive transfer of immune splenocytes from immunized inbred A/J mice conferred a significant level (p less than 0.001) of protection against subsequent challenge. Phenotypic analysis in outbred as well as two different strains of inbred mice (A/J and C57BL/6) demonstrated that CD8+ T cells are selectively stimulated by this immunization protocol. T cell depletion studies using specific mAb directed at either CD3+ or CD8+ T cell phenotype, followed by adoptive transfer, failed to confer protective immunity, whereas CD4+ depletion had no effect. These cytotoxic CD8+ T cells produced high titers of both IFN-gamma and IL-2. Moreover, these CD8+ T cells were directly parasiticidal against radiolabeled extracellular T. gondii, further supporting the critical immune function of these p30 Ag-specific CD8+ T cells in host immunity against T. gondii infection.

Adjuvants, Immunologic↗
PubMed1991

Possible regulation of the in vitro assembly of bovine brain tubulin by the bovine thioredoxin system.

Microtubule assembly in vitro and in vivo is highly sensitive to a variety of sulfhydryl-reactive reagents, raising the question of the possible existence of a physiological sulfhydryl-mediated system for regulating microtubule assembly. However, the specific reagents which have previously been used to inhibit microtubule assembly in vitro are either nonphysiological or, if physiological, effective only at concentrations much higher than their physiological ones. Because of reports of association in vivo between microtubules and the sulfhydryl-reactive proteins thioredoxin and thioredoxin reductase, we decided to examine the interaction in vitro between microtubules and the thioredoxin system, comprising thioredoxin, thioredoxin reductase and NADPH. At pH 6.8, both the mammalian and the Escherichia coli thioredoxin systems inhibited microtubule assembly by 4-35% (19 +/- 9%) by reducing one intra-subunit disulfide bond in the tubulin dimer. The thioredoxin-reducible disulfide of the tubulin dimer remains protected from thioredoxin in the assembled microtubules. Thioredoxin or thioredoxin reductase alone, or together in the absence of NADPH, were incapable of either reducing tubulin or inhibiting microtubule assembly. Microtubules formed from reduced tubulin were found to be stable and morphologically identical to those obtained from native tubulin dimers. Since the components of the thioredoxin system were used at concentrations similar to their physiological ones, our results suggest a potential role of the thioredoxin system in regulation of microtubule assembly in vivo.

Animals↗
PubMed1991

Changes in hypothalamic monoamine oxidase activity in relation to 17 beta-estradiol and clomiphene citrate treatments in the teleost Channa punctatus during three seasons.

Hypothalamic monoamine oxidase (MAO) activity displayed biphasic responses to administration of estradiol-17 beta (E2) depending on season. Lower doses of 0.05 and 0.10 microgram/g BW of E2 had enhanced enzyme activity and higher doses (0.5 and 1.0 microgram/g BW) depressed it significantly in preparatory season. In prespawning phase, the lowest dose of 0.05 microgram/g BW of E2 was stimulatory and the higher doses were inhibitory to MAO activity. In postspawning season, lower doses (0.05 and 0.10 microgram/g BW) of E2 failed to elicit any significant change in enzyme activity whereas the higher doses (0.5 and 1.0 microgram/g BW) elevated it significantly. Administration of clomid (5, 10, and 20 micrograms/g BW) inhibited MAO activity in a dose-dependent manner during all the three seasons. The results show that E2 may participate in feedback control of gonadotropin secretion by modifying monoaminergic activity at the level of oxidative deamination by MAO.

Animals↗
PubMed1991

Pineal-gonadal relationship in the teleost Channa punctatus (Bloch): evidence for possible involvement of hypothalamic serotonergic system.

In Channa punctatus, pinealectomy results in an accelerated growth (stimulatory effect) of the ovary in preparatory phase (March), but had no significant effect in prespawning (May-June) or postspawning (September) phases. Administration of melatonin (25 micrograms/fish or 0.4 mg/kg BW at 2-day intervals for 30 days) inhibits the stimulatory effect of a long photoperiod and high temperature (16L:8D; 22 +/- 1 degree C) on the ovary in the early preparatory phase (February). In the late preparatory phase (April), the administration of both melatonin (0.5 mg/kg BW at 2-day intervals) and parachlorophenylalanine (pCPA, a serotonin synthesis blocker; 100 mg/kg BW at 3-day intervals) for 30 days inhibits ovarian activity in comparison to that of the saline-treated control group. In the dose-response study, greater than or equal to 0.5 mg/kg BW of melatonin induced a significant increase in hypothalamic 5-HT content and greater than or equal to 10 mg/kg BW of pCPA decreased it significantly. In the time-course study, melatonin (2.0 mg/kg BW) elevates the 5-HT content significantly after an hr of the injection and maintained it up to 48 hrs. The administration of pCPA (200 mg/kg BW) had significantly inhibited the 5-HT content which was sustained for 72 hr. In another study, a single injection of melatonin (0.5 mg/kg BW) increased the 5-HT content significantly. A single injection of pCPA (100 mg/kg BW) decreased significantly both the content and activity of 5-HT. It is inferred that hypothalamic 5-HT may play a central role in photosexual mechanisms and mediate long photoperiodic effects on neuroendocrine-reproductive axis.

Animals↗
PubMed1990

Selective osmotic effect on diffusion of plasma membrane lipids in maize protoplasts.

Osmotic levels in the range typically used during plant protoplast isolation and incubation were investigated with regard to effects on the lateral diffusion of lipid probes in the plasma membrane. The lateral diffusion coefficient of a fluorescent sterol probe in the plasma membrane of maize (Zea mays L.) root protoplasts in a medium containing 0.45 M mannitol was 4 times faster than when the medium contained 0.9 M mannitol. The lateral diffusion coefficient of a fluorescent phospholipid probe, however, did not change over this range of mannitol concentrations. Similar diffusion characteristics were observed when the medium contained trehalose instead of mannitol. Slower lateral diffusion of the sterol probe at higher osmolality was also observed when KCl/CaCl2-based osmotic media were used with protoplasts isolated by a mechanical, rather than by an enzymic, method. Extraction and quantitation of total lipids from protoplasts showed that both the phospholipid and sterol contents per protoplast decreased with increasing osmolality, while the sterol/phospholipid ratio increased. These results demonstrate that osmotic stress induces selective changes in both the composition and biophysical properties of plant membranes.

Cell Membrane↗
PubMed1990

Effects of season, pinealectomy, and blinding, alone and in combination, on hypothalamic monoaminergic activity in the teleost Channa punctatus (Bloch).

In Channa punctatus, day-night variations in hypothalamic 5-HT (serotonin) and monoamine oxidase (MAO) activity were noticed in preparatory but not other phases (prespawning and postspawning) of the annual reproductive cycle. Hypothalamic MAO, 5-HT, and norepinephrine (NE) activity was found to be high in the prespawning phase and low in the postspawning phase. Dopamine (DA) activity, on the other hand, was high in the postspawning season and low in the prespawning phase. Pinealectomy caused season-dependent effects on hypothalamic monoaminergic activity, with a significant increase in serotonergic activity and a significant reduction in MAO activity at midscotophase during the preparatory phase (March) but not in the prespawning (May-June) or postspawning (September) phases. Hypothalamic catecholaminergic (CA) activity was not influenced by pinealectomy during any of the seasons. To determine whether or not the photoperiodic influences on daily variations of 5-HT and MAO in the preparatory phase are mediated via pineal and/or lateral eyes, fish were pinealectomized and/or blinded in January, when there is no rhythm, and sacrificed in February, when a day-night variation normally sets in. The day-night difference in 5-HT content and activity and MAO activity was not abolished by pinealectomy or blinding alone; but the combination (pinealectomy + blinding) obliterated the daily variation only in 5-HT content and in MAO activity. However, pinealectomy and blinding, alone or in combination, caused a significant elevation of 5-HT activity (not its level) and a significant decrease in MAO activity at midscotophase, with the combination having an additive effect. Hypothalamic CA content or activity was not affected by these regimes. The results show that photoperiodic influence on the daily pattern of 5-HT and MAO activity is mediated through and by the interaction of the pineal and lateral eyes.

Animals↗
PubMed1990

Induction of antigen-specific human cytotoxic T cells by Toxoplasma gondii.

To further the understanding of the role of T cells in immunity to the parasite Toxoplasma gondii, antigen-specific T cell clones were generated using peripheral blood mononuclear cells from seropositive individuals. Whole parasites were used to stimulate a proliferative expansion of antigen-reactive cells, followed by limiting dilution cloning in the presence of irradiated, autologous PBMC and rIL-2. Three parasite antigen-specific T cell clones expressing the CD3+ phenotype were selected for further characterization. Phenotypic analysis with monoclonal antibodies revealed two clones reactive with CD8 (RTg1 and RTg3) while the other (RTg2) phenotyped as CD4+, CD8-. When tested in a proliferation assay using a panel of different T. gondii proteins, clone RTg1 reacted with a single large protein (Mr greater than 180,000) as well as smaller components (less than 12,000), clone RTg2 reacted with a protein of Mr = 28,000 and clone RTg3 reacted with a protein of 116,000 plus smaller components (less than 12,000). Only the 28,000 = Mr antigen recognized by RTg2 was reactive on Western blot with autologous donor antisera. All three clones produced IFN-gamma and IL-2 in varying amounts upon antigenic stimulation in the presence of irradiated APC. Moreover, one clone RTg1, exhibited direct parasite cytotoxicity, inhibiting extracellular T. gondii by greater than 70% when incubated at an effector/target ratio of 40:1. This clone was alpha, beta TCR heterodimer positive and exerted its cytotoxic parasiticidal activity in the apparent absence of MHC restriction. The results provide evidence for the existence of circulating antigen-specific cytotoxic T cells in normal humans who are toxoplasma antibody seropositive.

Animals↗
PubMed1990

Prevalence of cryptosporidium in Rawalpindi/Islamabad a comparison of saline, iodine and stained preparations.

Three hundred stool samples for routine ova and parasite examination were examined as saline and iodine wet preparations, the results were then compared with the Zeihl-Neelsen-modified Acid Fast staining (MAFS), as the oocysts are acid fast in character. Examination with saline preparation was unable to detect any oocyst, while iodine preparation yielded three oocyst and stained smears with MAFS revealed five positive stool samples.

Animals↗
PubMed1988

Induction of antigen-specific parasiticidal cytotoxic T cell splenocytes by a major membrane protein (P30) of Toxoplasma gondii.

Infection with Toxoplasma gondii has become a major cause of morbidity in patients with AIDS. To investigate the mechanisms responsible for immune responses to toxoplasma Ag we used a highly purified membrane protein (P30) of T. gondii to stimulate an in vitro Ag-specific cytotoxic T cell response. P30 immune mouse splenocytes reduced extracellular T. gondii plaque-forming units by more than 50% when incubated at an E/T ratio of 10:1 or greater. By using a [3H]uracil radioisotope release assay, the effect of the immune splenocytes was determined to be a direct parasite lytic mechanism. The immune splenocytes were P30 Ag specific and of the Thy 1.2, Lyt2,3+ (CD4-, CD8+) phenotype, specific for mouse cytotoxic T cells. Opsonization of the parasites with monoclonal P30-reactive mAb did not enhance parasiticidal activity. Culture supernatants obtained during the 2-h cytotoxic assay were not parasiticidal, and anti-asialo-GM1 antibody plus C did not destroy the parasiticidal activity of the P30 responder cells. Accordingly, we have identified an Ag-specific subset of CD4-, CD8+, P30 responder T cells that are directly parasiticidal to extracellular T. gondii, and that exhibit cytotoxicity independent of antibody opsonization, lymphokine secretion, NK cell activity, and, apparently, MHC involvement as well.

Animals↗
PubMed1988

Native-like intermediate on the folding pathway of Escherichia coli succinyl-CoA synthetase.

The transition between the native and denatured states of the tetrameric succinyl-CoA synthetase from Escherichia coli has been investigated by circular dichroism, fluorescence spectroscopy, cross-linking by glutaraldehyde and activity measurements. At pH 7.4 and 25 degrees C, both denaturation of succinyl-CoA synthetase by guanidine hydrochloride and refolding of the denatured enzyme have been characterized as reversible reactions. In the presence of its substrate ATP, the denatured enzyme could be successfully reconstituted into the active enzyme with a yield of 71-100%. Kinetically, reacquisition of secondary structure by the denatured enzyme was rapid and occurred within 1 min after refolding was initiated. On the other hand, its reactivation was a slow process which continued up to 25 min before 90% of the native activity could be restored. Both secondary and quaternary structures of the enzyme, reconstituted in the absence of ATP, were indistinguishable from those of the native enzyme but the renatured protein was catalytically inactive. This observation indicates the presence of catalytically inactive tetramer as an intermediate in the reconstitution process. The reconstituted protein could be reactivated by ATP even 10 min after the reacquisition of the native secondary structure by the refolding protein. However, reactivation of the protein by ATP 60 min after the regain of secondary structure was significantly less, suggesting that rapid refolding and reassociation of the monomers into a native-like tetramer and reactivation of the tetramer are sequential events; the latter involving slow and small conformational rearrangements in the refolded enzyme that are likely to be associated with phosphorylation.

Circular Dichroism↗
PubMed1988

Production of gamma interferon by cultured human lymphocytes stimulated with a purified membrane protein (P30) from Toxoplasma gondii.

Purified P30, the principal iodinatable membrane protein of Toxoplasma gondii, induced proliferation of peripheral blood mononuclear cells from seropositive individuals but not from seronegative individuals. Culture supernatants from stimulated cells of seropositive individuals blocked the growth of T. gondii in human fibroblasts, whereas those from antibody-negative individuals failed to do so. The anti-toxoplasmic effect of culture supernatants correlated with the induction of indoleamine 2,3-dioxygenase and the destruction of tryptophan, as previously described for fibroblasts treated with recombinant gamma interferon (IFN-gamma). The anti-toxoplasmic effect was blocked by monoclonal antibody to IFN-gamma. The protective effect correlated with the amount of IFN-gamma in the culture supernatant, as measured by inhibition of viral CPE. Thus, the level of IFN-gamma appears to be an important immune factor in protection against toxoplasmosis in humans.

Animals↗
PubMed1988

Seasonal and daily variations in hypothalamic monoamine levels and monoamine oxidase activity in the teleost Channa punctatus (Bloch).

In Channa punctatus, a significant daily variation in hypothalamic 5-HT level and monoamine oxidase (MAO) activity was noticed in preparatory phase (February), but not in prespawning (May) or postspawning (November) phases. Hypothalamic dopamine (DA) and noradrenaline (NA), on the other hand, showed marked daily variation in their levels during all the three seasons with peak values in the photophase. The overall activity of MAO (mean +/- SEM on 24-hr period) increased from November to May through February, whereas the 5-HT content which was high in November decreased during February and May. The NA and DA levels were low in November and February and high in May. The catecholamine (CA) content and MAO activity increased with increasing photoperiod and temperature which is indicative of an enhanced CA metabolism.

Animals↗