Biomedical subjects

A Siddiqui

Publications and source records attributed to A Siddiqui.

At least 109 records · Page 6Linked to original sources

PubMed1990

Low-dose preoperative aspirin therapy, postoperative blood loss, and transfusion requirements.

The effects of three low-dose regimens of preoperative aspirin therapy on postoperative blood loss, transfusion requirements, and length of hospital stay were recorded in a prospective cohort study of 202 patients undergoing elective coronary artery bypass grafting. One hundred one patients had been prescribed daily aspirin by the referring cardiologist (44 at 75 mg, 28 at 150 mg, and 29 at 300 mg); the remaining 101 patients who had not been prescribed aspirin acted as a control group. A median postoperative blood loss of 870 mL in the control group was increased by 280 mL in the 75-mg aspirin group (p less than 0.001), by 490 mL in the 150-mg aspirin group (p less than 0.001), and by 230 mL in the 300-mg aspirin group (p = 0.03). The median requirement for blood transfusion of 2 U red blood cell concentrates in the control group was increased by 2 U in the 75-mg aspirin group (p less than 0.001), 2 U in the 150-mg aspirin group (p less than 0.001), and 1 U in the 300-mg aspirin group (p = 0.05). Hemostatic "packs" (fresh frozen plasma, platelets, and cryoprecipitate) were required in 20 patients in the aspirin groups as compared with 5 in the control group (p less than 0.01 by chi 2 test). The mean postoperative hospital stay was 8 days for all groups. Regular daily low-dose aspirin therapy produces significant increases in postoperative blood loss, resulting in a substantial increase in blood transfusion and hemostatic pack requirements, but does not prolong postoperative hospital stay.

Aspirin↗

PubMed1990

Cell surface expression of hepatitis B surface and core antigens in transfected rat fibroblast cell lines.

Hepatocellular necrosis during hepatitis B virus infection is hypothesized to result from host immune responses against either hepatitis B surface antigen or hepatitis B core antigen expressed on the surface membrane of infected hepatocytes. To study the capacity of hepatitis B deoxyribonucleic acid to induce membrane expression of either hepatitis B surface antigen or hepatitis B core antigen in vitro, we assessed transfected rat fibroblast cell lines by indirect immunofluorescence. Rat fibroblasts were transfected with plasmid vectors containing the natural promoters, native enhancer, and uninterrupted sequences of either the Pre S/S gene or core gene. Resulting cell lines produced hepatitis B surface antigen and hepatitis B core antigen/hepatitis B e antigen, respectively. Immunofluorescence microscopy or flow cytometry showed that hepatitis B surface antigen and hepatitis B core antigen were expressed in a granular pattern in the surface membrane of transfected cells. We conclude that surface membrane expression of both hepatitis B surface antigen and hepatitis B core antigen is an intrinsic consequence of expression of either the Pre S/S or core gene.

Animals↗

PubMed1990

Multiple liver-specific factors bind to the hepatitis B virus core/pregenomic promoter: trans-activation and repression by CCAAT/enhancer binding protein.

The human hepatitis B virus (HBV) is a hepatotropic virus that replicates through an RNA intermediate referred to as the pregenome. The promoter that directs the synthesis of the pregenome and several other transcripts with heterogeneous 5' ends is of particular interest because of its role in regulating key functions during the viral life cycle. We have examined the liver-specific characteristics of this promoter by DNA-protein interactions and by demonstrating the in vivo function of the promoter using the luciferase reporter gene expression system. The DNA-protein interactions in this region appear to be almost entirely liver-specific. Among these, a liver-specific nuclear factor, CCAAT/enhancer binding protein, binds to at least five sites on this promoter. Transient cotransfection experiments using CCAAT/enhancer binding protein expression vectors and the core promoter in the context of either the native hepatitis B virus genome or the luciferase reporter gene demonstrate that CCAAT/enhancer binding protein at low concentration modestly activates expression from the core promoter but represses at high concentration.

Animals↗

PubMed1990

Trans-activation of HLA-DR gene by hepatitis B virus X gene product.

Hepatocellular injury during hepatitis B virus (HBV) infection has been postulated to result from a human leukocyte antigen (HLA)-restricted T-lymphocyte host immune response against HBV antigens. Although HLA expression is enhanced in the presence of hepatic inflammation, whether HBV itself can induce HLA expression on infected hepatocytes is unknown. In this study, we demonstrate the induction of HLA-DR expression on human hepatoma cell lines transfected with HBV DNA sequences. The HBV X gene alone was capable of inducing HLA-DR expression. This induction correlated with elevated HLA-DR RNA, and this resulted directly from transcriptional trans-activation of the HLA-DR gene by the HBV X protein. These studies suggest that the HBV X protein can regulate the expression of HLA-DR and thus raise the possibility of participation by the X gene in the immunopathogenesis of HBV infection.

Antigens, Viral↗

PubMed1990

The efficacy of tongue resection in treatment of symptomatic macroglossia in the child.

Nine patients with symptomatic macroglossia, 3 with Beckwith-Wiedemann syndrome, 3 with Down's syndrome, and 3 with lymphatic malformation of the tongue, were evaluated for postoperative improvement following partial glossectomy. Symptoms evaluated were speech intelligibility, oral competence at rest, drooling, and normal deglutition. All patients with Down's syndrome and Beckwith-Wiedemann syndrome were improved postoperatively. All patients with Down's syndrome exhibited improved deglutition and reduced drooling. Only 1 patient (33%), however, achieved oral competence. Speech was improved postoperatively in 2 patients (67%) with Down's syndrome. Patients with lymphatic malformation of the tongue did not exhibit consistent long-term improvement postoperatively, suggesting that conservative management of macroglossia in this patient group may be indicated.

Adolescent↗

PubMed1990

Assessment of ventricular function in infants and children. Response to dobutamine infusion.

A method to assess ventricular functional reserve in infants and children unable to perform dynamic exercise was evaluated. Left ventricular ejection fraction was measured by radionuclide angiocardiography at rest and during infusion of dobutamine in 5-15 micrograms/kg/min dosages. The only side effects noted were arrhythmias in two patients, who had similar ectopy documented previously. Group left ventricular ejection fraction increased from 0.40 +/- 0.21 to a maximum of 0.49 +/- 0.24 (P less than 0.001). Left ventricular ejection fraction at 5 micrograms/kg/min (0.44 +/- 0.23) was not statistically different from that measured during infusion of dobutamine at 10 micrograms/kg/min (0.48 +/- 0.24). The six patients receiving anthracyclines, who had relatively low function at rest and increased function during dobutamine administration, were continued on their anticancer regimen without deteriorating in condition. Absolute values of left ventricular ejection fraction correlated best with the prognosis in patients with idiopathic dilated cardiomyopathy or structural abnormalities; six out of ten patients whose left ventricular ejection fraction never rose above 0.40 have either died or are in transplant protocols. Thus, study of left ventricular function can be performed easily during dobutamine infusion, generating valuable prognostic information.

Antineoplastic Agents↗

PubMed1990

Hepatitis B virus X protein produced in Escherichia coli is biologically functional.

The hepatitis B virus X gene product trans activates transcription from a variety of viral and cellular regulatory elements. We expressed the complete, nonfused X protein in Escherichia coli and showed it to be active in trans activating a human immunodeficiency virus long terminal repeat-linked chloramphenicol acetyltransferase reporter gene.

Chloramphenicol O-Acetyltransferase↗

PubMed1990

Post-mortem changes of neurotransmitter concentrations in the rat brain regions.

Using High Performance Liquid Chromatography coupled with electrochemical detection the post-mortem stability of noradrenaline (NA), dopamine (DA), serotonin (5-HT) and 5-hydroxy indole acetic acid (5-HIAA) were examined in the rat hypothalamus, amygdala, cerebral cortex, cerebellum and corpus striatum over an 8 hour time period. Changes in concentrations of the different neurotransmitters were less than it might be expected. The significant changes were: a. A fall in NA levels in the cerebral cortex by 4 hours and in the hypothalamus at 8 hours. b. A reduction in DA concentrations in the corpus striatum at 8 hours but a two fold rise of levels in the hypothalamus at 1 and 2 hours. c. A four-fold increase in 5-HT concentrations in the amygdala throughout the 8 hours studied. The results indicate that for comparative studies on post-mortem brain tissue correction factors should be employed to take into account differential changes in the concentrations of the various neurotransmitters.

Animals↗

PubMed1990

Effect of mancozeb on hydrolytic metabolism of xenobiotics.

The present investigation reports the effect of chronic oral administration of mancozeb, a fungicide, on hepatic microsomal carboxylesterases/amidases or B-esterases responsible for hydrolytic metabolism of aspirin (acetylsalicylic acid or ASA) at pH 5.5 and 7.4, 2-acetylaminofluorene (AAF), acetanilide and p-nitrophenylacetate (NPA) and cholinesterase in rat. Oral administration of mancozeb (250 mg/kg/day) for 30 days caused significant stimulation of ASA esterase I (pH 5.5), ASA esterase II (pH 7.4), AAF N-deacetylase and acetanilide N-deacetylase in liver. However, the activities of NPA esterase and cholinesterase remained unaffected. Evaluation of induction kinetics demonstrated that the pattern and magnitude of responses of these microsomal hydrolases to mancozeb treatment for 7 days were comparable to those obtained after treatment for 30 days. The activities of hydrolases were not altered in animals killed 4 hr after an oral dose of mancozeb. Mancozeb did not affect these hydrolases in vitro.

2-Acetylaminofluorene↗

PubMed1989

trans-activation of viral enhancers including long terminal repeat of the human immunodeficiency virus by the hepatitis B virus X protein.

Human hepatitis B virus contains an open reading frame designated X. We have investigated the trans-activating function of the hepatitis B virus X gene in regulating transcriptional control elements. In the HBV genome the major target for X trans-activation is the enhancer element. Further, the X protein stimulates several other viral promoters/enhancers including the long terminal repeats (LTR) of human retroviruses. One of the viral sequences studied in detail is the human immunodeficiency viral (HIV) LTR which is trans-activated by the X protein. Using mutational analysis of the HIV LTR, we show that the NF-kappa B sequences contained within the U3 region are involved in this stimulatory activity. Nuclear run-on analyses support the notion that X-mediated trans-activation occurs at the level of transcription.

DNA Mutational Analysis↗

PubMed1989

Negative regulation of the hepatitis B virus pre-S1 promoter by internal DNA sequences.

Expression of the surface antigen gene (S gene) of hepatitis B virus is directed by two distinct promoter elements with markedly different activities. The upstream (pre-S1) promoter produces a 2.4-kb transcript at very low levels while the downstream (pre-S2) promoter produces an approximately 2.1-kb transcript in relative abundance. We have constructed a series of internal deletion mutants to analyze differential regulation of the two S gene promoters. We show here that expression directed by the pre-S1 promoter is negatively regulated by DNA sequences containing the downstream pre-S2 promoter region. Nuclear run-on analysis indicates this down-regulation to be at the level of transcription. Furthermore, promoter repression does not appear to be due to products of the S gene region. Deletion mutagenesis studies have permitted the localization of a 61-bp region that may be involved in the apparent down-regulation of the pre-S1 promoter. These results suggest the use of an unusual regulatory mechanism by a dipromoter gene in which an active internal promoter may preclude efficient use of an upstream promoter.

Base Sequence↗

PubMed1989

Interactions between nuclear factors and the hepatitis B virus enhancer.

We have previously established that the tissue-specific activity of the hepatitis B virus (HBV) enhancer is mediated by trans-acting cellular factors. Here we have studied in vitro the interactions between the HBV enhancer DNA and cellular factors present in nuclear extracts from both liver and nonliver cell types. The results presented in this study imply the involvement of several distinct, ubiquitous, and liver-specific cellular factors with the HBV enhancer. Sequence analysis of the binding sites for these proteins on HBV DNA showed homologies to sequence motifs known to bind other previously characterized and purified transcription factors including CAAT/enhancer-binding protein. Thus, all of these binding sites may function in concert to activate liver-specific transcription of HBV genes from their respective promoters.

Base Sequence↗

PubMed1989

Enzyme immunoassay in the diagnosis of Chlamydia trachomatis infections in diverse patient groups.

An enzyme immunoassay (EIA) in parallel with cell culture was used to investigate the extent of infections due to Chlamydia trachomatis. EIA reactive confirmed in cell culture was taken as positive. C. trachomatis was found in 6 (26.0%) of 23 men with symptomatic non-gonococcal urethritis (NGU), ten (17.2%) of 58 symptom-free males and in three of 4 with postgonococcal urethritis. Among 106 asymptomatic pregnant women studied the incidence of C. trachomatis was 8.5% while a higher incidence (16.7%) was found in those with symptoms. C. trachomatis positivity in asymptomatic and symptomatic post-natal screening were 11.4% and 7.7%. Of 43 symptomatic non-pregnant females investigated, 7 (16.3%) were found to be positive for C. trachomatis. Of 3 women with PID, 2 (66.7%) harboured C. trachomatis in their cervix while in another 29 infertile women, C. trachomatis was positive in 3 (8.1%). Contraceptives appeared to have an effect on the chlamydial positivity. Comparative testing of EIA with the standard cell culture method in this study indicate EIA as a suitable alternative for the definitive diagnosis of chlamydial infection in high prevalence settings and with caution in low prevalence settings.

Antigens, Bacterial↗

PubMed1988

Site-directed mutagenesis of the reactive center (serine 394) of antithrombin III.

Human antithrombin III (AT) shares significant sequence homology and a common inhibitory mechanism with the serine protease inhibitor (serpin) superfamily. AT has a reactive site in which the P1 residue is primarily responsible for protease specificity. The P1' residue, almost invariably serine, is critical in the inactive natural variant AT-Denver, which has a leucine substitution in that position (Stephens, A.W., Thalley, B.S., and Hirs, C.H.W. (1987) J. Biol. Chem. 262, 1044-1048). In the present study site-directed mutagenesis was used to generate eight variants with altered P1' residues. All were secreted efficiently by COS cells transiently transfected with the AT cDNA in a eukaryotic shuttle vector. All variants also bound heparin as effectively as wild-type AT. Variants were grouped into three categories with respect to thrombin-AT complex formation: 1) no detectable inhibitory activity (proline, methionine); 2) low activity (cysteine, valine, leucine); and 3) near normal activity (glycine, alanine, threonine). The leucine variant, which is in the low activity group, exhibited the same physical and functional properties as AT-Denver. We conclude that the serine hydroxyl is not critical for functional activity and that there is a side chain size optimum which is modulated by hydrophobic effects.

Amino Acid Sequence↗

PubMed1988

The hepatitis B virus enhancer modulates transcription of the hepatitis B virus surface antigen gene from an internal location.

We have used a transient expression system to examine the influence of the hepatitis B virus enhancer on the transcription of the hepatitis B virus surface antigen (HBsAg) gene. This enhancer is located 3' to HBsAg coding sequences but is contained within the mature HBsAg transcripts. Removal of the enhancer resulted in a significant reduction in HBsAg gene expression in liver cells. By using an in vitro nuclear run-on assay, this effect was determined to be at the level of transcription. We have further noted that HBsAg expression can be restored by the reinsertion of the enhancer sequences in the enhancerless vector. However, this reconstitution of HBsAg expression appears to depend on location of the enhancer with regard to the HBsAg promoters.

Animals↗

PubMed1988

Regional differences in the catecholamine content of the rat brain: effects of neonatal castration and androgenization.

The long-term organizational influence of perinatal androgen manipulations upon the central noradrenergic and dopaminergic systems was investigated in the rat. Males were castrated or sham-operated upon within 24 h of birth. Newborn females received either five daily sc injections of 100 micrograms of testosterone in oil, or vehicle only. Groups of animals were then decapitated at ages ranging from 12 to 180 days. Levels of noradrenaline and dopamine were measured in six brain regions using high-performance liquid chromatography with electrochemical detection. The results confirm that the noradrenaline and dopamine content of the brain are affected by perinatal androgen exposure and show that brain catecholamine levels alter during the first six months of life, in some areas merely increasing, but in others rising and later falling. Sex differences in catecholamine levels may be apparent at particular ages, but the direction of the difference is frequently reversed as the animals grow older. In the hypothalamus and amygdala broadly similar patterns of time-related changes in catecholamine levels were observed. However, although in other regions (cerebral cortex, cerebellum, corpus striatum and corpora quadrigemina) sex differences in the noradrenaline and dopamine content were frequently detected, on most occasions they were not affected in any recognizable fashion by steroid manipulation in the newborn. Once adult patterns of gonadotropin secretion and behaviour became established in early adulthood, a functional relationship between brain catecholamine levels and the early effects of testosterone is no longer so apparent. Nevertheless, it would seem that testosterone, acting during the neonatal period, tends to accelerate the fall in brain amine levels that occurs as the animals grow older.

Age Factors↗

PubMed1987

Expression of functionally active human antithrombin III.

Human antithrombin III cDNA was cloned into an expression vector suitable for transient expression in COS cells. Upon transfection COS cells secreted a single immunoreactive 58-kDa protein. Quantitation of secretion levels by ELISA indicated that at 44 hr posttransfection cells were secreting 48 +/- 5 ng of antithrombin III per 10(6) cells per 24 hr. Heparin-agarose chromatography resulted in the elution of the COS-derived protein as a broad band between 0.3 and 1.0 M NaCl. 35S-labeled medium from transfected cells reacted with human thrombin (1.5 ng/ml) in the absence of heparin. In 40 min, greater than 80% of the immunoreactive material was found as a higher molecular weight species, consistent with stoichiometric covalent complex formation. In a two-stage chromogenic thrombin inactivation assay, under pseudo-first-order conditions, at 16 nM antithrombin III the t1/2 was 74 min and 50 min for plasma and COS cell-derived antithrombin III, respectively, in the absence of heparin. In the presence of 17.4 nM high-affinity heparin, the t1/2 was 5.2 min and 2.2 min, respectively.

Animals↗

PubMed1987

Expression of the hepatitis B virus X gene in mammalian cells.

The hepatitis B virus (HBV) DNA contains an open reading frame (ORF) designated X, which has the capacity to encode a protein of 16,560 Da (subtype adw). Such a protein has not been identified in either HBV particles or infected human livers, and therefore its role in the viral life cycle remains unknown. We report here the expression of the HBV X ORF in cultured cells using recombinant vectors. A protein of 16 kDa was identified by means of an antiserum prepared against a synthetic peptide and with human antisera from hepatitis B patients as well as those with hepatocellular carcinoma. Cell fractionation and immunofluorescence studies suggest a probable association with cytoskeletal components. Our studies further located a promoter sequence upstream of the X ORF, which directs the transcription of a 0.7- to 0.8-kilobase X-specific RNA in transfected human hepatoma cells.

Acetyltransferases↗