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Biomedical subjects

S Rahman

Publications and source records attributed to S Rahman.

At least 163 records · Page 9Linked to original sources

On the induction of protective responses in Salmonella typhimurium strain TA1535/pSK1002 by UVA (365 nm).

Exposure to UVA (365 nm) led to growth delay, loss of viability and inhibition of 3H-thymidine incorporation into the cells of Salmonella typhimurium strain TA1535 containing multiple copies of a plasmid pSK1002 carrying a umuC'-'lacZ fusion gene. Ultraviolet-A induced umu gene expression, as monitored by the estimation of beta-galactosidase, in a linear fluence-dependent manner. The induction of umu gene expression increased with the increase of postirradiation incubation period of the cells in the LB-ampicillin (LBA) medium at 37 degrees C and leveled off from 2 h onward. The induction of gene expression depended on concomitant protein synthesis and represented the induction of the SOS response in the particular S. typhimurium cells used. The exposure to low fluences (sublethal) of UVA also led to the induction of an adaptive response in the same bacterial cells, which made them resistant to subsequent challenge by a much higher fluence of the same radiation. The adaptive response, as monitored by the assays of viability and beta-galactosidase units, increased with the period of exposure to sublethal fluences of UVA, attained a maximum at the UVA exposure of 4.5 kJ/m2 (15 min) and thereafter gradually decreased with further increase of UVA exposure period. Modulation studies involving D2O, LBA growth medium, different scavengers of free radicals and quenchers of activated oxygen species indicated the involvement of both hydroxyl free radicals and singlet oxygen in the UVA-induced umu gene expression.

Bacterial Proteins↗

Open versus laparoscopic cholecystectomy: a comparison of postoperative temperature.

Laparoscopic cholecystectomy has been shown to allow better postoperative pulmonary function than open cholecystectomy, with less incidence of lung atelectasis. As atelectasis following abdominal surgery is responsible for most febrile episodes in the first 48 h, it was postulated that with minimally invasive surgery there may be a parallel improvement in the incidence of postoperative fever. This study was designed to evaluate this hypothesis. Seventy-eight patients were randomly divided into two groups. Thirty-eight had an open cholecystectomy and 40 underwent the laparoscopic approach. Twenty-one of the 38 patients (55%) following open cholecystectomy had early febrile episodes compared with only 6 of the 40 (15%) in the laparoscopic cholecystectomy group. We concluded that there was a lower incidence of febrile episodes following laparoscopic cholecystectomy and suggest that this was related to improved pulmonary function and minimal surgical trauma.

Adult↗

Preferential antagonism of the interactions of the integrin alpha IIb beta 3 with immobilized glycoprotein ligands by snake-venom RGD (Arg-Gly-Asp) proteins. Evidence supporting a functional role for the amino acid residues flanking the tripeptide RGD in determining the inhibitory properties of snake-venom RGD proteins.

The inhibitory properties of a panel of snake-venom-derived RGD (Arg-Gly-Asp) proteins, including the disintegrins kistrin, elegantin and albolabrin, and the neurotoxin homologue dendroaspin, were investigated in a platelet-adhesion assay using three immobilized ligands of the glycoprotein IIb-IIIa complex (alpha IIb beta 3), namely fibrinogen, fibronectin and von Willebrand factor (vWF). The snake-venom proteins preferentially inhibited the adhesion of ADP-treated platelets to one or more of the immobilized ligands. Kistrin and dendroaspin exhibited similar inhibitory characteristics, abrogating platelet adhesion to fibrinogen and vWF at nanomolar concentrations, but poorly inhibiting adhesion to fibronectin. Kistrin and dendroaspin share little overall amino-acid-sequence identity, but a considerable level of sequence similarity exists around the RGD tripeptide. Synthetic cyclic peptides corresponding to these regions of kistrin and dendroaspin inhibited platelet adhesion to both fibrinogen and fibronectin with approximately equal potency, but were 100-fold weaker antagonists of the interactions of the alpha IIb beta 3 complex with fibrinogen than their parent proteins. The disintegrins elegantin and albolabrin, which share approx. 60% overall amino-acid-sequence similarity with kistrin but have different residues around the RGD tripeptide, exhibited different antagonistic preferences. Elegantin inhibited platelet adhesion to immobilized vWF and fibronectin, but was significantly less effective at disrupting adhesion to fibrinogen. Albolabrin selectively inhibited platelet adhesion to immobilized vWF and was less effective with fibrinogen and fibronectin as adhesive ligands. In contrast with the behaviour of these venom proteins, the adhesion of ADP-treated platelets to immobilized fibrinogen, fibronectin and vWF was inhibited non-selectively by a range of monoclonal antibodies with specificity for the alpha IIb beta 3 complex. These observations, therefore, define antagonistic preferences in this panel of venom proteins towards the interactions of the alpha IIb beta 3 complex with three immobilized glycoprotein ligands.

Amino Acid Sequence↗

Cloning of a wheat 15-kDa grain softness protein (GSP). GSP is a mixture of puroindoline-like polypeptides.

The wheat starch 15-kDa protein (called grain softness protein or GSP) consists of a major polypeptide and several minor polypeptides. An antiserum raised against GSP was used to screen a wheat cDNA library. A cDNA family encoding approximately 15-kDa proteins that included a heptapeptide sequence previously isolated from protease digests of GSP was identified. A partial cDNA was used in a prokaryotic expression system to produce a fusion protein which reacted strongly against the original anti-GSP serum. A new antiserum raised against the fusion protein produced a weak reaction against a 15-kDa polypeptide extracted from wheat seeds. The results suggest that the proteins encoded by the cDNA family form a minor component of the mixture of 15-kDa polypeptides defined as GSP. RNA complementary to the cDNAs could be extracted from both soft and hard wheat grains from about half-way through grain filling. The encoded proteins are novel members of the 2S superfamily of seed proteins, a diverse family of proteins which maintain a characteristic framework of cysteine residues. The deduced proteins show the highest similarity to the oat 16-kDa avenin and to wheat puroindoline (a lipid-binding 15-kDa protein from wheat). Review of previously published data shows that puroindoline is also closely related to the major polypeptide of GSP, suggesting that the lipid-binding properties of GSP polypeptides may influence grain softness.

Amino Acid Sequence↗

Production and disposal of medium-chain fatty acids in children with medium-chain acyl-CoA dehydrogenase deficiency.

The effect of fasting on plasma concentrations of fatty acids has been determined in four children with medium-chain acyl-CoA dehydrogenase (MCAD) deficiency. In addition, the in vivo rate of octanoate oxidation was measured, using [1-13C]octanoate. In the three older children (1.5-11.2 years), fasting for up to 18 h stimulated lipolysis, as reflected by the increasing concentration of free fatty acids, but with little rise in concentrations of medium-chain fatty acids, octanoate, decanoate and cis-4-decenoate. In an infant (0.5 year), lipolysis was greater and was accompanied by rising concentrations of medium-chain fatty acids. After 13.5 h there was a rapid increase in the concentration of decanoate and cis-4-decenoate. The calculated in vivo rate of octanoate oxidation was substantial in all patients studied (6.4-13.1 mumol/kg per h) despite very low MCAD activity in vitro. It is concluded that under basal conditions the in vivo oxidation rate of medium-chain fatty acids is near normal in the four children studied with MCAD deficiency.

Acyl-CoA Dehydrogenase↗

Immunohistological expression of p53 in primary pT1 transitional cell bladder cancer in relation to tumour progression.

OBJECTIVE: To determine whether p53 expression is a marker of tumour progression in superficially invasive (pT1) transitional cell carcinoma of the bladder. PATIENTS AND METHODS: The immunohistochemical status of the p53 protein in 28 pT1 primary bladder cancers was determined on frozen tissue and archival paraffin block sections using three primary antibodies (CM-1, PAB1801 and D07). The findings were compared with the patients' progress. All the patients, except for those who died during the course of the study, were followed up by check cystoscopy for a minimum of 2 years. RESULTS: Immediately adjacent frozen sections stained identically in 10 of 16 cases for CM-1 and PAB1801. For paraffin sections, identical staining patterns were seen for PAB1801 and D07 in 21 of 26 sections. However, inter- and intra-tumour staining for p53 was very variable, even with the same antibody. The heterogeneity of p53 positive cell distribution in the tumours indicates potential for significant sampling errors if random sections are chosen as representative of p53 status. CONCLUSION: The p53 status of the primary tumours did not relate to patient outcome. The results obtained do not support the use of immunohistological p53 expression as a discriminating prognostic indicator in pT1 transitional cell carcinoma of the bladder.

Biomarkers, Tumor↗

Myo-inositol reduces serotonin (5-HT2) receptor induced homologous and heterologous desensitization.

The effect of myo-inositol was examined on 5-HT2 receptor mediated facilitation of NMDA depolarization of rat neocortical neurons in vitro. Myo-inositol (1-10 mM) potentiated the 5-HT facilitation, the potentiation increasing linearly with log 5-HT concentration. Myo-inositol also eliminated 5-HT induced heterologous desensitization of muscarinic and alpha 1-adrenergic receptor mediated facilitation. Our findings suggest that 5-HT induced homologous and heterologous desensitization results in part from depleting phosphoinositide substrate.

Animals↗

Synthetic RGD peptides derived from the adhesive domains of snake-venom proteins: evaluation as inhibitors of platelet aggregation.

Synthetic peptides based on the RGD domains of the potent platelet aggregation inhibitors kistrin and dendroaspin were generated. The 13-amino-acid peptides were synthesized as dicysteinyl linear and disulphide cyclic forms. In platelet-aggregation studies, the cyclic peptides showed 3-fold better inhibition than their linear equivalents and approx. 100-fold greater potency than synthetic linear RGDS peptides derived from fibronectin. An amino acid substitution, Asp10-->Ala, in the kistrin-based peptide gave a 4-fold decrease in potency in the linear peptide, but produced a 2-fold elevation in the inhibitory activity of the cyclic form, generating a peptide of potency comparable with that of the parent protein.

Adenosine Diphosphate↗

Identification of multiple glucocorticoid receptor binding sites in the rat osteocalcin gene promoter.

The biosynthesis of osteocalcin (OC), a bone-specific, noncollagenous protein, is stringently regulated during differentiation of the osteoblast phenotype. Glucocorticoids, and also 1,25(OH)2D3, mediate the developmental regulation of OC gene transcription. In this study, we established that the -1097 to +23 promoter (pOCZCat) of the rat OC gene confers glucocorticoid responsiveness to both basal and vitamin D-induced OC expression. The presence of multiple glucocorticoid receptor (GR) binding sites in the proximal rat OC gene promoter was determined by the combined use of DNase I footprinting, dimethyl sulfate fingerprinting, and gel mobility shift analysis with glucocorticoid receptor protein. One glucocorticoid receptor binding element (GRE) resides immediately downstream of the TATA box (-16 to -1). In vivo activity was established by cotransfection of ROS 17/2.8 osteosarcoma cells with an OC-CAT construct in the presence of cloned GRE sequences (wild type or mutant) as competitors. A putative second, less protected GR binding site is located further upstream in the OC gene basal promoter within the region overlapping the TATA box. This is in direct contrast to the organization of GREs in the human OC proximal promoter wherein GR binding at the upstream GRE overlapping the TATA is stronger than at the downstream GRE. In addition, we detected sequence-specific binding of GR protein to another basal promoter element, the OC box (-99 to -76), which contains a central CCAAT motif. The presence of multiple GR binding sites in the rat OC gene proximal promoter indicates that regulation of basal and vitamin D-enhanced transcription by glucocorticoids may involve the integrated activities of multiple, independent GREs.

Animals↗

Multiple mechanisms of serotonin 5-HT2 receptor desensitization.

Desensitization of serotonin 5-HT2 receptor-mediated enhancement of the N-methyl-D-aspartate (NMDA) depolarization was studied in rat cortical neurons. Serotonin and (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) induced long term desensitization. Staurosporine, a nonspecific protein kinase C inhibitor, potentiated the serotonin and DOI facilitation, suggesting acute desensitization was operative. In the case of DOI, long term desensitization was prevented by staurosporine. Activators of protein kinase C abolished the serotonin facilitation, an action prevented by staurosporine. Concanavalin A potentiated the facilitation at 100 microM, but not 30 microM serotonin, suggesting these receptors undergo dose dependent internalization. Calmodulin antagonists prevent long term desensitization induced by serotonin. The depolarization induced by NMDA alone was not altered by staurosporine, protein kinase C activators, concanavalin A or calmodulin antagonists. Serotonin at 100 microM, but not 30 microM, induced heterologous desensitization of phenylephrine and carbachol induced facilitation of the NMDA depolarization. We conclude that serotonin 5-HT2 receptors both induce and undergo several forms of desensitization.

Alkaloids↗

Activation of 5-HT2 receptors facilitates depolarization of neocortical neurons by N-methyl-D-aspartate.

The interaction between serotonin and excitatory amino acid agonists at rat neocortical neurons was investigated using the grease-gap recording method. Depolarization evoked by 50 microM N-methyl-D-aspartate was dose dependently facilitated by serotonin (5-HT) (1 to 100 microM) giving a bell-shaped dose-response curve with maximum enhancement at 30 microM. In contrast, quisqualate and kainate depolarizations were not enhanced. Subnanomolar concentrations of methysergide, ritanserin and spiperone, but not ICS 205-930, attenuated the 5-HT enhancement, compatible with 5-HT2, but not 5-HT1 or 5-HT3 receptor subtype involvement. Enhancement was observed with 5-HT2 receptor agonists, whereas 5-HT1 receptor subtype agonists had either no effect (1B and 1C) or reduced (1A) the N-methyl-D-aspartate depolarization. Scopolamine and prazosin reduced the N-methyl-D-aspartate depolarization and blocked facilitation induced by carbachol and phenylephrine, but not that due to 5-HT. Tetrodotoxin reduced the N-methyl-D-aspartate depolarization, but the facilitation by 5-HT persisted. Activators of protein kinase C (phorbol diacetate and 1-oleoyl-2-acetyl-sn-glycerol) did not mimic the serotonin facilitation. We conclude that serotonin enhances N-methyl-D-aspartate depolarization of rat cortical neurons through activation of 5-HT2 receptors, however the cellular mechanism underlying the facilitation remains to be established.

Animals↗

Assembly of hepatic gap junctions. Topography and distribution of connexin 32 in intracellular and plasma membranes determined using sequence-specific antibodies.

The subcellular distribution in rat liver and the topography in intracellular and plasma membranes of connexin 32, a major protein component of gap junctions, was studied using sequence-specific anti-peptide antibodies generated to extracellular and intracellular domains of the protein. The distribution of connexin 32 in liver analyzed using SDS-polyacrylamide gel electrophoresis and Western blotting showed the relative protein levels in the subcellular fractions to be: lateral plasma membranes > Golgi membranes > sinusoidal plasma membranes > lysosomes. Low amounts of connexin 32 were detected in microsomes, endosomes, and bile canalicular plasma membranes. Six highly conserved cysteine residues are located in the amino acid sequences comprising the two extracellular loops of all connexins thus far isolated, and these loops are positioned to extend the channel in the lipid bilayers across the intercellular region of the gap junction. In the present work, the intramolecular disulfide bonds linking the extracellular loops in gap junctions were shown to be present in connexins located in plasma membranes, Golgi, and a microsomal fraction, and it was concluded that the disulfide linkages were formed in the endoplasmic reticulum. In addition, immature configurations of connexin 32, probably occurring during membrane insertion, were detected in liver microsomal fractions. The results contribute to charting of the biogenetic routes followed by connexins in hepatocytes and the general mechanisms of gap junction assembly.

Amino Acid Sequence↗

The effect of interleukin-1 on cytokine gene expression in cultured human articular chondrocytes analyzed by messenger RNA phenotyping.

OBJECTIVE: To investigate the pattern of cytokine gene expression in human articular chondrocytes in culture in response to interleukin-1 beta (IL-1 beta). The effect of serum and variations in culture conditions was also studied. METHODS: Messenger RNA was extracted from cells, reverse-transcribed to complementary DNA, and amplified by the polymerase chain reaction (PCR), using specific oligonucleotide primers. The PCR products were validated by restriction analysis with specific enzymes and by Southern blot analysis. RESULTS: In cultured articular chondrocytes, IL-1 beta, IL-1 alpha, granulocyte colony-stimulating factor (CSF), and granulocyte-macrophage CSF cytokine genes were expressed only after induction by IL-1 beta. However, IL-6, IL-8, and macrophage CSF genes were expressed constitutively. The expression of IL-1 beta was dose and time dependent. CONCLUSION: Using PCR, it was possible to demonstrate gene expression for several cytokines in human articular chondrocytes in culture. It was evident that some cytokine genes were expressed constitutively and some were inducible by IL-1 beta.

Base Sequence↗

A five-year clinical evaluation of Norplant contraceptive subdermal implants in Bangladeshi acceptors.

A non-comparative study of the Norplant contraceptive subdermal implant system was conducted in Dhaka, Bangladesh. The study was designed to evaluate the contraceptive efficacy, safety and overall acceptability of Norplant implants. Six hundred women were enrolled in the clinical trial, which began in 1985 at three study sites. Follow-up visits were scheduled at 1, 3, and 6 months after Norplant implant insertion, and every six months thereafter until removal or at the end of five years. There were no post-insertion pregnancies during Norplant implant use in this study. After five years of Norplant implant use, there was no clinically significant change in body weight, systolic or diastolic blood pressure. Less than 3% of the women ever reported having any significant medical problems such as migraine, respiratory or cardiac problems during the study. The gross cumulative continuation rate was 41.2 per 100 women at the end of five years. The two most frequently reported reasons for discontinuation during the study were menstrual problems and desired pregnancy. Of the women who completed the five-year user satisfaction questionnaire, the majority of the women (86.3%) planned to continue using contraception after study completion. Of these women, approximately one-third said they planned to use another Norplant set. The findings presented suggest that the Norplant system is a highly effective, safe and acceptable method among Bangladeshi women.

Adolescent↗

Primary actinomycotic endocarditis: case report and review.

Endocarditis due to Actinomyces species for which a portal of entry cannot be identified is extremely rare. We present a case of primary endocarditis due to Actinomyces israelii with an unknown portal of entry and review seven similar cases reported in the literature since 1939. The disease predominantly affects males. Clinical features are similar to those of bacterial endocarditis due to other organisms. Penicillin remains the drug of choice for treatment of this condition. The optimal duration of treatment has not yet been determined; however, with early diagnosis and appropriate antimicrobial therapy, the prognosis is good.

Actinomyces↗

Multiple copies of the bone-specific osteocalcin gene in mouse and rat.

The osteocalcin (OC) gene was initially described as a single copy gene encoding the bone specific vitamin K dependent and vitamin D regulated protein. We report here the presence of multiple copies of the gene in mouse and rat. Southern blot analysis and restriction mapping of genomic DNA from several strains of mice indicated the presence of at least three copies of the OC coding sequence within a 19 kb fragment. Two closely linked OC genes contain the proximal promoter region with intact coding sequences. The third potential OC gene includes a 3.5 kb insert between an OC promoter-like region and a coding region that has several amino acid substitutions distributed among functional domains when compared with the normal gene. The 940 nucleotides upstream of the modified coding region lack the well defined 5' regulatory elements that support basal and hormone-responsive transcriptional control. In rats either one or more OC genes were observed in different strains or in Sprague Dawley rats obtained from different suppliers.

Amino Acid Sequence↗