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Biomedical subjects

M Collins

Publications and source records attributed to M Collins.

At least 19 recordsLinked to original sources
PubMed1992

The abolition of collagen gene expression in SV40-transformed fibroblasts is associated with trans-acting factor switching.

The goal of this study was to determine whether alpha 2(1) procollagen gene expression is modulated by positive or negative trans-acting DNA-binding proteins. Previous studies have shown that a clone of SV40-transformed WI-38 fibroblasts (SVWI-38) does not produce any alpha 2(1) procollagen mRNA (Parker et al (1989), J. Biol Chem. 264, 7147-7152). In order to elucidate the mechanism(s) responsible for such inactivation, we have examined the activity of a transfected wild type COL1A2 promoter in SVWI-38 cells. A set of 5' promoter deletions was linked to the chloramphenicol acetyltransferase (CAT) gene and transfected into SVWI-38 and other cell lines expression type I collagen. The resulting CAT assays confirmed the importance of several upstream regions for promoter activity and documented the decreased transcriptional activity from an exogenous COL1A2 promoter in the SVWI-38 cell line. Competition experiments with an excess of COL1A2 promoter DNA fragment and a constant amount of COL1A2/CAT construct displayed a linear relationship between excess COL1A2 fragment and CAT activity in SVWI-38 cells, suggesting the involvement of a titratable negative effector. Electrophoretic mobility shift assays revealed the presence of a specific DNA-protein complex which was present in SVWI-38 cells and almost absent in control fibroblasts. Methylation interference analysis mapped the region of binding of this factor between nucleotides -80 and -72, relative to the transcription start site. Thus the data presented provide strong evidence for the existence of a negative trans-acting factor that may play a role in the repression of COL1A2 expression in SVWI-38 fibroblasts.

Base Sequence↗
PubMed1992

Characterization of a novel T lymphocyte protein which binds to a site related to steroid/thyroid hormone receptor response elements in the negative regulatory sequence of the human immunodeficiency virus long terminal repeat.

We have previously identified a T lymphocyte protein which binds to a site within the LTR of the human immunodeficiency virus type 1 (HIV-1) and exerts an inhibitory effect on virus gene expression. The palindromic site (site B) recognized by this protein is related to the palindromic binding sites of members of the steroid/thyroid hormone receptor family. Here we characterize the T cell protein binding to this site as a 100 kD protein which is most abundant in T cells and which binds to site B as a 200 kD complex. This protein is distinct from other members of the steroid/thyroid hormone receptor family including the COUP protein which has a closely related DNA binding specificity.

Base Sequence↗
PubMed1992

Glutamate differentially inhibits the expression of class II MHC antigens on astrocytes and microglia.

MHC molecules are required for Ag recognition by T cells. Inasmuch as cells in the central nervous system do not express MHC constitutively, appearance of MHC, in inflammatory and degenerative diseases of the brain, may indicate local Ag presentation and subsequent immune response. Although both astrocytes and microglia are capable of class II MHC expression in vitro, in vivo studies failed to show the presence of significant amounts of class II on astrocytes compared to microglia. Our study is designed to clarify possible regulatory mechanisms that can explain the differences in inducibility of class II MHC between astrocytes and microglia in vivo. Using dissociated rat brain cell cultures, we have found that glutamate, an excitatory neurotransmitter, exerted a profound inhibitory effect on IFN-gamma-induced expression of class II on astrocytes, but not on microglia. Both glutamate and norepinephrine, a neurotransmitter previously reported to down-regulate class II on astrocytes, inhibited the induction of class II on astrocytes by eliminating accumulation of class II MHC mRNA. The kinetics of class II mRNA induction by IFN-gamma in the presence of glutamate suggested that glutamate may act as a transcriptional inhibitor. It is likely that class II induction on astrocytes in vivo may be selectively down-regulated by neurotransmitters such as glutamate and norepinephrine.

Animals↗
PubMed1992

A flexible, efficient, checkerboard immunoblot system for the detection and semiquantitation of specific antinuclear antibodies.

In this paper an uncomplicated method for the simultaneous detection and semiquantitation of 11 of the 12 commonly studied antinuclear antibodies (ANA) in a single run is described. This new application of checkerboard immunoblotting (CBIB) is based upon available technology and employs purified antigens which can be either purchased or produced in-house. CBIB requires no electronic instrument, can be formatted to meet the needs of the user, is rapidly performed, and has acceptable labor and materials costs. Data on the use of the method to examine available reference antisera is presented. CBIB has also proven practical for the clinical study of 18 sera, at two dilutions per membrane, for each set of specific antinuclear antibodies, also at two or more dilutions.

Antibodies, Antinuclear↗
PubMed1992

Effect of lithium on rat embryos in culture: growth, development, compartmental distribution and lack of a protective effect of inositol.

Lithium chloride (LiCl) was tested at various concentrations (50, 100, 150 and 200 micrograms/ml) using a rat whole-embryo culture system beginning on gestation day 9.5 (headfold stage) in order to establish a concentration-response relationship. Open neural tubes--as described in former publications (Tesh 1988)--were not induced by lithium. A significant reduction of embryonic growth and development occurred at the lowest concentration tested (50 micrograms/ml). Clear-cut dysmorphogenetic effects (absence of the eye cup, kink in the spinal anlage, "bleb" at the rostral head region) occurred at a concentration of 150 micrograms/ml LiCl. LiCl concentrations in the embryo, visceral yolk sac, exocoelomic fluid and medium were determined after the embryos had been cultured for 48 h in the presence of a moderately embryotoxic dose of LiCl (3.5 mEq/l or 150 micrograms/ml). Medium supplementation with myo-inositol in different concentrations was ineffective in antagonizing the embryotoxicity induced by LiCl.

Animals↗
PubMed1992

Suppressor T cells: some answers but more questions.

Many properties of suppressor T cells and the antigen-binding factors derived from them have evaded molecular genetic definition. Here, Martin Dorf and colleagues discuss recent data in the context of a growing awareness of the molecules and principles involved.

Animals↗
PubMed1992

Rothmund-Thomson syndrome: review of the world literature.

Rothmund-Thomson syndrome is a rare, inherited disorder characterized by poikilodermatous skin changes that appear in infancy. The inheritance is autosomal recessive. Patients exhibit variable features including skeletal abnormalities, juvenile cataracts, and a higher than expected incidence of malignancy. This extensive review of the world literature clarifies aspects of the inheritance, incidence of characteristic features, and malignant potential of this disease. Insight into its origin is provided through a review of the clinical and in vitro studies of endocrine function and possible DNA repair abnormalities.

Adolescent↗
PubMed1992

Prostaglandin E2 gel induction of patients with a prior low transverse cesarean section.

To determine safety and efficacy of induction with prostaglandin E2 gel, we compared the outcome of 25 patients (study group) with an unfavorable cervix, a medical indication for delivery, and one prior low cervical transverse cesarean section to 56 patients (comparison group) with one prior low cervical transverse cesarean section and spontaneous labor. We placed 1 mg of prostaglandin E2 in gel intracervically in the 25 study patients. Common indications for delivery were: diabetes, post dates, and preeclampsia. Although most labor and delivery variables were similar, the study group had a longer mean latent phase (14.2 +/- 13.8 versus 7.3 +/- 3.7 hours: p less than 0.002), but had a shorter mean length of active phase (4.0 +/- 3.5 versus 5.7 +/- 3.0 hours; p less than 0.02). None of the patients in either group had a dehiscence of the uterine scar, nor rupture of the uterus. Both groups had a similar cesarean section rate. Since from the few reported, nonrandomized studies it appears that prostaglandin E2 gel use in patients with a prior low cervical transverse cesarean section may be useful and relatively safe, it may be time to attempt randomized trials of prostaglandin E2 gel versus oxytocin for induction of patients with a prior low cervical transverse cesarean section, unfavorable cervix, and a medical indication for delivery.

Adult↗
PubMed1992

Visceral leishmaniasis in the BALB/c mouse: antimony tissue disposition and parasite suppression after the administration of free stibogluconate.

BALB/c mice with an acute or chronic Leishmania donovani infection were treated with intravenous sodium stibogluconate solution and the parasite suppressions determined in the spleen, liver and femur bone marrow. Antimony concentrations in these and other tissues were determined by hydride generation-atomic absorption spectrophotometry. There was little correlation between tissue antimony levels one hour after treatment and drug efficacy. It would appear that the peak tissue antimony concentration achieved soon after dosing, rather than the lower concentrations which are readily sustained in most tissues, is the most important factor in the antileishmanial activity of stibogluconate. A high peak antimony concentration occurred in the liver, where parasites were significantly suppressed, and was not observed in the two other sites of infection, where the parasites were apparently less susceptible to stibogluconate therapy.

Acute Disease↗
PubMed1992

Controlled trial using computerized feedback to improve physicians' diagnostic judgments.

The goal of this study was to test an innovative method to improve physicians' diagnostic judgments by integrating the use of a computer program (employing cognitive feedback to teach a clinical rule that predicts the probability of streptococcal pharyngitis), a traditional lecture, and periodic disease-prevalence reports. In a controlled trial using pre- and postintervention measures involving 885 patients, the authors compared the effects of the integrated method on the diagnostic judgments of seven experienced physicians at a university health service (from 1982 to 1985) with the effects of the lecture alone on the judgments of seven experienced physicians at a different university health service (1986 to 1987). The integrated method significantly improved the quality of the physicians' judgments as measured by calibration curves and Brier scores, and increased the level of agreement between the physicians' judgments and those made by the clinical prediction rule. The lecture alone produced less improvement in the quality of the physicians' judgments, and decreased the level of agreement with the rule. The authors conclude that this method, based on cognitive psychology, is a promising educational tool.

Computer-Assisted Instruction↗
PubMed1992

Human papillomavirus type 16 E5 gene stimulates the transforming activity of the epidermal growth factor receptor.

We have until recently made several unsuccessful attempts to assign any activity to the human papillomavirus type 16 (HPV-16) E5 gene product. However, studies with the bovine papilloma virus 1 (BPV-1) E5 protein indicated an interaction with the epidermal growth factor receptor (EGFR). In light of the overall similarity between the HPV and BPV E5 proteins we attempted to determine whether the HPV-16 E5 gene had any common activity. In cells expressing high levels of EGFR plus HPV-16 E5 we found a dramatically increased proliferative activity in soft-agar assays in the presence of EGF. The specificity of this activity was monitored by the addition of other mitogenic agents. The phorbol ester phorbol 12-myristate 13-acetate (PMA) had no effect on the E5-containing cells, although insulin weakly stimulated their growth in soft agar. Further analysis revealed the same number of EGF receptors were present on the E5-containing cells as on the control cells, although the E5 cells were more sensitive to lower concentrations of EGF. These results imply that E5 is amplifying the mitogenic signals from the EGFR in an as yet unknown manner, but which may form the basis of interactions with a variety of growth factor receptors. This report brings to three the number of transforming genes encoded by HPV-16.

3T3 Cells↗
PubMed1992

Iron and nutritional status of toddlers.

The iron and nutritional status of 22 clinically well and normal toddlers, aged from one to four years inclusive was determined. Mean energy intake was normal. Protein, fibre and ascorbic acid intakes were high. Total iron intake was low in 73%, and available iron was low in 96%. The percentage availability of the total iron intake was 6.98 +/- 1.2, which is below the 10% assumption of iron availability based on consumption of the recommended daily allowances. Mean haemoglobin was with in the normal range. Low normal serum ferritin suggested poor iron stores. Anthropometric measurements were normal. Although the toddlers; intake of most dietary parameters was normal, the finding of poor iron intake and stores is of concern given recent reports of the role of iron in weight gain and psychomotor development.

Child Nutritional Physiological Phenomena↗
PubMed1991

An exon 5-deleted mRNA encodes a functional interleukin 2 receptor alpha-subunit.

Two polypeptides are involved in interleukin 2 binding: a low-affinity receptor of 55 kD (IL2-R alpha) and an intermediate affinity component of 75 kD (IL2-R beta). We describe the cloning by the Polymerase Chain Reaction of the coding region of IL2-R alpha from a human T-cell lymphoma cell line. One clone presented a 72-bp deletion that precisely corresponds to exon 5. The deleted form and the normal IL2-R alpha cDNA were expressed CHO cells. Stable transfected cellular clones were compared for their immunoreactivity to monoclonal antibodies directed against IL2-R alpha and for their ability to bind radiolabeled IL2. The presence or absence of the protein region encoded by exon 5 did not modify the IL2-binding capacity of the receptor.

Amino Acid Sequence↗
PubMed1991

T-cell receptor alpha chain plays a critical role in antigen-specific suppressor cell function.

Antigen-specific suppressor T-cell hybridomas release soluble suppressor factors (TsF) in the supernatant that modulate both in vivo delayed-type hypersensitivity and in vitro plaque-forming cell responses in an antigen-specific manner. To study the relationship between the T-cell receptor (TcR) and TsF, we developed a series of TcR alpha- or TcR beta- expression variants from suppressor T-cell hybridomas that expressed the CD3-TcR alpha/beta complex. We demonstrate that loss of TcR alpha but not TcR beta mRNA was accompanied by the concomitant loss of suppressor bioactivity. Homologous transfection of TcR alpha cDNA into a TcR alpha- beta+ clone reconstituted both CD3-TcR expression and suppressor function. Furthermore, suppressor activity from TcR beta- variants was specifically absorbed by antigen and anti-TcR alpha antibodies, but not by anti-CD3 or anti-TcR beta affinity columns. These data directly establish a role for the TcR alpha chain in suppressor T-cell function and suggest that the TcR alpha chain is part of the antigen-specific TsF molecule.

Animals↗
PubMed1991

Molecular cloning in Lactobacillus helveticus by plasmid pSA3::pVA797 co-integrate formation and conjugal transfer.

A gene encoding beta-glucanase activity from Bacillus amyloliquefaciens was subcloned in both orientations into plasmid shuttle vector pSA3. In only one orientation could a co-integrate be generated with the conjugative plasmid pVA797. The plasmid co-integrate was conjugated into Lactobacillus helveticus strain CNRZ450, where it was stably maintained without antibiotic selection and exhibited beta-glucanase activity. This method of introducing cloned DNA into thermophilic lactobacilli will facilitate the study of heterologous gene expression in non-transformable species.

Cloning, Molecular↗
PubMed1991

A comparison of fasting serum amino acid profiles of young and elderly subjects.

The fasting serum amino acid profile in 37 healthy young women and men (30-35 years) was compared with the fasting profile in 30 institutionalized elderly women and men (80-89 years), an ambulatory, self-fed senior-residence group. Levels of serum lysine, leucine, methionine, valine, and total essential amino acids were significantly lower in the older group than in the younger group; however, citrulline and hydroxyproline were significantly higher in the older group compared to the younger group. Histidine, threonine, tryptophan, and the ratio of tryptophan to large neutral amino acids (isoleucine, leucine, phenylalanine, tyrosine, and valine) were also generally lower in the older group than in the younger group, while the difference due to age was more pronounced in the females compared to males. The essential/nonessential amino acid ratio was lower in females compared to males.

Adult↗
PubMed1991

In-patient withdrawal from long-term benzodiazepine use.

Twenty-one patients completed an intensive in-patient and out-patient group-based programme, to achieve withdrawal from long-term benzodiazepine dependence. By 6-month follow-up (post-withdrawal), eight patients (38%) had achieved a 'good' outcome. Generally, patients demonstrated moderate levels of psychiatric morbidity and depression in the pre-withdrawal phase; a diagnosis of a depressive disorder being associated with either an 'intermediate' or 'poor' outcome in comparison to patients receiving a diagnosis of anxiety disorder, who showed a predominantly 'good' outcome. Two patients committed suicide some time after withdrawal. Additionally, older patients fared less well than their younger counterparts. We conclude that maintenance benzodiazepine prescription, rather than withdrawal, may be the preferred treatment for some dependent patients. Furthermore, more data are required regarding the association, and causality, between depression and benzodiazepine withdrawal in long-term users.

Adult↗