Biomedical subjects

F Martin

Publications and source records attributed to F Martin.

At least 217 records · Page 12Linked to original sources

PubMed1996

YY1 and NF1 both activate the human p53 promoter by alternatively binding to a composite element, and YY1 and E1A cooperate to amplify p53 promoter activity.

A novel transcription factor binding element in the human p53 gene promoter has been characterized. It lies about 100 bp upstream of the major reported start site for human p53 gene transcription. On the basis of DNase I footprinting studies, electromobility shift assay patterns, sequence specificity of binding, the binding pattern of purified transcription factors, effects of specific antibodies, and methylation interference analysis we have identified the site as a composite element which can bind both YY1 and NF1 in an independent and mutually exclusive manner. The site is conserved in the human, rat, and mouse p53 promoters. The occupancy of the site varies in a tissue-specific manner. It binds principally YY1 in nuclear extracts of rat testis and spleen and NF1 in extracts of liver and prostate. This may facilitate tissue-specific control of p53 gene expression. When HeLa cells were transiently transfected with human p53 promoter-chloramphenicol acetyltransferase reporter constructs, a mutation in this composite element which disabled YY1 and NF1 binding caused a mean 64% reduction in basal p53 promoter activity. From mutations which selectively impaired YY1 or NF1 binding and the overexpression of YY1 or NF1 in HeLa cells we concluded that both YY1 and NF1 function as activators when bound to this site. In transient cotransfections E1A could induce the activity of the p53 promoter to a high level; 12S E1A was threefold as efficient as 13S E1A in this activity, and YY1 bound to the composite element was shown to mediate 55% of this induction. Overexpressed YY1 was shown to be able to synergistically activate the p53 promoter with E1A when not specifically bound to DNA. Deletion of an N-terminal domain of E1A, known to be required for direct E1A-YY1 interaction and E1A effects mediated through transcriptional activator p300, blocked the E1A induction of p53 promoter activity.

Animals↗

PubMed1996

Cloning of the bovine activin receptor type II gene (ACVR2) and mapping to chromosome 2 (BTA2).

The cDNA for the bovine activin receptor type II (ACVR2) gene has been cloned and sequenced. It encodes a protein of 513 amino acids which is highly homologous (approximately 98% identity) to the human, mouse, and rat proteins. Using PCR analysis of bovine x hamster somatic cell lines, the ACVR2 gene was mapped to syntenic group U17, which has been localised to bovine chromosome 2. Comparative mapping has shown that the genes within U17 are also in a syntenic group on the long arm of human and sheep chromosome 2, as well as on mouse chromosome 1. This group of genes represents an evolutionarily conserved mammalian chromosomal segment. Genotyping a highly polymorphic microsatellite locus, (AT)4(GT)9(AT)11, found within an intron of this gene confirmed the localisation by linking the ACVR2-associated microsatellite to the region of chromosome 2 flanked by CSSM42 and TGLA226. This gene locus, which has the characteristics of a type I and type II mapping locus, represents the first localisation of this gene in any species to date.

Activin Receptors↗

PubMed1996

The effect of heavy social drinking on recall and event-related potentials.

OBJECTIVE: It has been suggested that alcohol-related brain impairments progress on a continuum from subtle deficits in social drinkers through to end-state Korsakoff syndrome. The aim of the present study was to investigate whether heavy levels of social drinking have an adverse effect on sober cognitive functioning in young nonalcoholic adults. METHOD: Event-related potentials (ERPs) were recorded from heavy ( > 200 gm/week) and light ( < 20 gm/week) male social drinkers under the effects of a pharmacological challenge (lorazepam-ATIVAN) and a placebo. The heavy (n = 14) and light (n = 14) social drinkers completed a free-recall task with a rare word probability of .30. RESULTS: Analysis of the P300 component of the ERP to recalled and not-recalled words found that P300 amplitude in heavy social drinkers (HSDs) was reduced following placebo compared to the light social drinkers (LSDs). Lorazepam produced a distinctive pattern of anterograde memory deficits in both groups and reduced P300 amplitude to rare words in the LSDs compared to both the placebo treatment and the HSDs. CONCLUSIONS: The differences in central nervous system functioning evident between HSDs and LSDs were reflected in ERP deviations in both the presence and absence of lorazepam. Even though no definite statements can be made because of the small sample size, the results appear to indicate that HSDs have an impairment in their information-processing ability in the absence of an acute inhibitory agent (e.g., alcohol or a benzodiazepine) as well as a differential response to the depressant effects of lorazepam.

Adult↗

PubMed1996

Diadenosine polyphosphates. A novel class of glucose-induced intracellular messengers in the pancreatic beta-cell.

Diadenosine polyphosphates are a group of low-weight compounds that increase after exposure to a wide variety of oxidants and have been suggested to act as "alarmones," alerting the cell to the onset of metabolic stress. We demonstrate here that glucose at concentrations that induce insulin release produce a 30- to 70-fold increase in the concentration of diadenosine triphosphate (Ap3A) and tetraphosphate (Ap4A) in beta-cells. Furthermore, Ap3A and Ap4A, at the concentrations found in glucose-stimulated cells, are effective inhibitors of the ATP-regulated K+ channels when applied to the intracellular side of excised membrane patches from cultured beta-cells. We suggest that Ap3A and Ap4A act as second messengers mediating a glucose-induced blockade of the pancreatic beta-cell ATP-regulated potassium channel.

Adenosine Triphosphate↗

PubMed1996

Zinc causes an apparent increase in rhodopsin phosphorylation.

PURPOSE: Rhodopsin is a zinc-binding protein. We investigated the effect of low concentrations of zinc on the initial phosphorylation of rhodopsin. METHODS: Dark-adapted bovine rod outer segments (ROS) were incubated with (gamma 32P) ATP and 5 mM magnesium in the presence and absence of micromolar amounts of zinc. The ROS were exposed to light to initiate phosphorylation under conditions which allow only limited initial phosphorylation. RESULTS: We found that zinc enhanced the rhodopsin phosphorylation apparent on autoradiographies by several fold. Phosphorylation reactions conducted in the presence of potent phospho-opsin phosphatase inhibitors show a comparable zinc-enhanced phosphorylation of rhodopsin. Under our reaction conditions, ROS membranes also appear more red upon initial exposure to light when zinc is present. CONCLUSIONS: Zinc can increase initial rhodopsin phosphorylation, apparently acting at the substrate rhodopsin and not at relevant phosphatases or rhodopsin kinase. How zinc binding to rhodopsin might increase its ability to serve as a substrate for phosphorylation is under investigation.

Animals↗

PubMed1996

Expression of fibronectin ED-A+ and ED-B+ isoforms by human and experimental colorectal cancer. Contribution of cancer cells and tumor-associated myofibroblasts.

Alternative splicing of primary fibronectin (FN) mRNA results in the synthesis of different isoforms. ED-A+ and ED-B+ FN isoforms are absent from plasma FN and are representative of cellular FN. Their expression was studied in human and rat normal colon, in human colorectal carcinomas, and in transplanted tumors derived from a chemically-induced rat colon cancer. In normal colon, only the ED-A+ FN isoform was expressed as a thin deposit between crypt colonocytes and pericryptal myofibroblasts. Conversely, heavy ED-A+ FN deposits and lighter ED-B+ FN expression were found in the stroma of colorectal tumors in association with myofibroblasts surrounding tumor glands. Some colonic cancer cells also contained intracellular FN isoform granules and expressed FN mRNA. Tumor-associated myofibroblasts and some cancer cell lines were able to synthesize and deposit extracellular ED-A+ and ED-B+ FN in vitro. FN isoform deposition by tumor-associated myofibroblasts was not modulated by colon cancer cell-conditioned medium, but was strongly enhanced when myofibroblasts were cultured on colon cancer cell extracellular matrix or on laminin. These results show that the ED-A+ and ED-B+ FN isoforms were overexpressed in colorectal cancer. Cancer cells can deposit these FN isoforms directly and also stimulate their deposition by tumor-associated myofibroblasts.

Actins↗

PubMed1996

Inflammatory cells infiltrating human colorectal carcinomas express HLA class II but not B7-1 and B7-2 costimulatory molecules of the T-cell activation.

Colon cancer cells express potentially immunogenic proteins but are not rejected by the immune system. To induce an effective immune response, antigenic peptides have to be presented to T lymphocytes by professional antigen-presenting cells in association with HLA class II molecules. Antigen-presenting cells also have to express B7 family molecules, B7-1 and B7-2, which deliver the costimulatory signals that are required to prevent T cell anergy. We studied B7-1 and B7-2 expression by the antigen-presenting cells that infiltrate colorectal cancer stroma. In 25 samples of colorectal carcinomas, a panel of monoclonal antibodies was used to label macrophages, dendritic cells, and T lymphocytes that infiltrate the tumor stroma and the morphologically normal distant mucosa. The expression of HLA class II and B7 molecules involved in T-cell activation was studied using specific monoclonal antibodies. Biopsy pieces from two patients with active Crohn's disease were used as controls. All of the samples were heavily infiltrated by macrophages and/or dendritic cells that strongly expressed HLA class II molecules. In contrast, antibodies to B7-1 and/or B7-2 stained no cells in 16 of the 25 samples of colorectal tumors and less than 1% of the inflammatory cells that infiltrated tumor stroma of the other nine tumor samples. B7 molecules were also poorly expressed by rare cells in the lamina propria of the morphologically normal colorectal mucosa. In contrast, many inflammatory cells that infiltrated the two Crohn's disease samples strongly expressed B7-1 and B7-2, especially in the granulomas. We conclude that most HLA class II+ inflammatory cells that infiltrate colorectal cancers do not express the B7-1 and B7-2 costimulatory molecules. This defect may contribute to the failure of the immune system to recognize tumor cells as antigenic.

Adenocarcinoma↗

PubMed1996

Selection of tRNA(Asp) amber suppressor mutants having alanine, arginine, glutamine, and lysine identity.

Elements that confer identity to a tRNA in the cellular environment, where all aminoacyl-tRNA synthetases are competing for substrates, may be delineated by in vivo experiments using suppressor tRNAs. Here we describe the selection of active Escherichia coli tRNAAsp amber mutants and analyze their identity. Starting from a library containing randomly mutated tRNA(CUA)Asp genes, we isolated four amber suppressors presenting either lysine, alanine, or glutamine activity. Two of them, presenting mainly alanine or lysine activity, were further submitted to a second round of mutagenesis selection in order to improve their efficiency of suppression. Eleven suppressors were isolated, each containing two or three mutations. Ten presented identities of the two parental mutants, whereas one had switched from lysine to arginine identity. Analysis of the different mutants revealed (or confirmed for some nucleotides) their role as positive and/or negative determinants in AlaRS, LysRS, and ArgRS recognition. More generally, it appears that tRNAAsp presents identity characteristics closely related to those of tRNALys, as well as a structural basis for acquiring alanine or arginine identity upon moderate mutational changes; these consist of addition or suppression of the corresponding positive or negative determinants, as well as tertiary interactions. Failure to isolate aspartic acid-inserting suppressors is probably due to elimination of the important G34 identity element and its replacement by an antideterminant when changing the anticodon of the tRNAAsp to the CUA triplet.

Alanine↗

PubMed1996

[Living conditions of prostitutes: consequences for the prevention of HIV infection].

Since 1990, several community-based interventions have been set up for populations of prostitutes in France with the principal objective of preventing HIV infection in prostitutes and their sexual partners. Field workers have suggested that extremely precarious living conditions are a major obstacle for the diffusion of prevention messages. A multidisciplinary working group thus set up a study in order to determine which living conditions could affect the adoption of behaviour at low risk for HIV infection. In May 1995, six of the seven teams conducting prevention actions among prostitutes in France used a short standardised questionnaire to collect information including type of housing, health insurance cover, physical aggressions, drug use. It was planned to collect data from all persons attending the drop-in centres. A total of 355 questionnaires were completed (sampling was exhaustive for 3 of the 6 teams). The population comprised women, men and transvestites. The median age was 28 years. Only 39% (135/348) of the subjects had access to health insurance. Approximately 50% (160/324) of the persons lived in precarious accommodation (hotel or no fixed address) and 33% (119/355) had been victims of physical aggressions during the 5 months preceding the study. Transvestites and young people had the most precarious living conditions. Despite the limits of this study due to the difficulty in carrying out a survey in this very marginalised population, the unique information collected may enable community health action strategies to be suitably adapted. In particular, it is important to improve partnership between field workers and existing social and health services, to improve access to better accommodation, health care and physical security while disseminating prevention messages and distributing condoms.

Adolescent↗

PubMed1996

[Arteriovenous malformation of the mandible. Apropos of a case in a 6-year-old child].

Arteriovenous malformations of the mandible are uncommon. Though characteristic clinical and radiological presentations are well described, management in the case of children is still a matter of debate. We report a mandibular angioma in a 6-year-old girl which was treated with superselective embolization. This method is the treatment of choice for mandibular arteriovenous malformations in children because of the low complication rate and the preservation of healthy tissue. When the malformation continues to develop despite repeated embolizations, surgical treatment may be necessary.

Arteriovenous Malformations↗

PubMed1995

Constitutive activation of mitogen-activated protein kinase-activated protein kinase 2 by mutation of phosphorylation sites and an A-helix motif.

A recently described downstream target of mitogen-activated protein kinases (MAPKs) is the MAPK-activated protein (MAPKAP) kinase 2 which has been shown to be responsible for small heat shock protein phosphorylation. We have analyzed the mechanism of MAPKAP kinase 2 activation by MAPK phosphorylation using a recombinant MAPKAP kinase 2-fusion protein, p44MAPK and p38/40MAPK in vitro and using an epitope-tagged MAPKAP kinase 2 in heat-shocked NIH 3T3 cells. It is demonstrated that, in addition to the known phosphorylation of the threonine residue carboxyl-terminal to the catalytic domain, Thr-317, activation of MAPKAP kinase 2 in vitro and in vivo is dependent on phosphorylation of a second threonine residue, Thr-205, which is located within the catalytic domain and which is highly conserved in several protein kinases. Constitutive activation of MAPKAP kinase 2 is obtained by replacement of both of these threonine residues by glutamic acid. A constitutively active form of MAPKAP kinase 2 is also obtained by deletion of a carboxyl-terminal region containing Thr-317 and the A-helix motif or by replacing the conserved residues of the A-helix. These data suggest a dual mechanism of MAPKAP kinase 2 activation by phosphorylation of Thr-205 inside the catalytic domain and by phosphorylation of Thr-317 outside the catalytic domain involving an autoinhibitory A-helix motif.

3T3 Cells↗

PubMed1995

Use of tension band wires in horses with fractures of the ulna: 22 cases (1980-1992).

Twenty-two horses with ulnar fractures were treated, using tension band wires alone or tension band wires in combination with pins or cortical bone screws. Age of the horses ranged from 2 weeks to 12 years (median, 4 months), and body weight ranged from 68 to 477 kg (median, 181 kg). Fractures were classified according to the Donecker and Bramlage ulnar fracture classification and included type 1-a (4 horses), type 1-b (4), type-2 (6), type-3 (1), type-4 (3), and type-5 (4), fractures. Tension band wires alone were used in 7 horses. Tension band wires were used in conjunction with unthreaded pins in 10 horses. In 3 horses, 5.5-mm cortical bone screws were inserted longitudinally instead of pins. A combination of a 5.5-mm cortical bone screw and a pin was used in 2 horses. In addition to pins and tension band wires, 4.5-mm cortical bone screws were placed in lag fashion to aid reduction of comminuted or oblique fractures in 7 of the 22 horses. Fractures healed in 18 (82%) horses. Four horses were euthanatized because of complications that included catastrophic failure of fixation during recovery from general anesthesia in 1 foal, septic arthritis and hyperextension of the contralateral metacarpophalangeal joint in 1 foal, and wound infection with partial disruption of repair in 2 horses. Nonfatal complications developed in 6 horses and included incision infection, partial wound dehiscence, carpal contraction, carpus varus of the contralateral forelimb, slight distraction of proximal fragments of the fractures, bent implants, and distal migration of pins. Long-term monitoring was performed on 17 horses.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthesia, General↗

PubMed1995

Development of the mouse B-cell repertoire.

The mouse B-cell repertoire early in ontogeny contains B cells with receptor immunoglobulins exhibiting high connectivity, multi/self-reactivity, and generally low affinity. This is due structurally to extensive restriction in the germline components used to generate the B-cell receptor. The selective pressure acting on the nascent repetoire has both negative and positive components as seen in our in vivo models. VH81X-bearing B cells from the VH81X transgenic mice (and probably from normal mice) are subject to self-selective pressure with two components: a positive one favoring a certain (self-reactive) specificity in the CD23-IgM+ population and a negative one preventing the entry of B cells with this specificity into the CD23+IgM+ compartment.

Animals↗

PubMed1995

Participation of the propeptide on procathepsin D activation of human peripheral lymphocytes and neutrophils.

Flow cytometry was used to test the role of the propeptide of procathepsin D in the activation of human peripheral lymphocytes and neutrophils. A selective inhibition of the activation was achieved by antibodies targeted against the propeptide. Synthetic peptide corresponding to the cathepsin D propeptide had a very similar effect as procathepsin D itself. The interaction of procathepsin D with the cells was blocked by the propeptide as monitored in experiments with fluorescently labeled procathepsin D. Our data indicate that procathepsin D activation of human neutrophils and leukocytes described earlier is mediated through the propeptide of the procathepsin D.

Cathepsin D↗

PubMed1995

Amino acid-induced [Ca2+]i oscillations in single mouse pancreatic islets of Langerhans.

1. The effects of amino acids on cytosolic free calcium concentration ([Ca2+]i) were measured, using fura-2 fluorescence imaging, in mouse pancreatic islets of Langerhans. 2. Slow [Ca2+]i oscillations appeared when isolated islets were incubated with a solution containing a mixture of amino acids and glucose at concentrations found in the plasma of fed animals. 3. In the presence of 11 mM glucose, alanine (5 mM) and arginine (10 mM) induced a transient rise in [Ca2+]i followed by an oscillatory pattern, while leucine (3 mM) and isoleucine (10 mM) triggered the appearance of slow [Ca2+]i oscillations. 4. Also in the presence of glucose (11 mM), tolbutamide (10 microM) increased the duration of the glucose-induced [Ca2+]i oscillations. While tolbutamide (10 microM) did not modify the leucine-induced slow oscillatory pattern, addition of diazoxide (10 microM) resulted in the gradual appearance of [Ca2+]i oscillations which resembled the glucose-induced fast oscillations. 5. Like stimulatory glucose concentrations (11 mM), glyceraldehyde (10 mM) induced fast oscillations of [Ca2+]i. 6. Fluoroacetate (2 mM) transformed leucine-induced slow [Ca2]i oscillations into fast [Ca2+]i oscillations. Iodoacetate (1 mM) completely inhibited any oscillatory pattern. 7. It is suggested that mitochondrially generated signals, derived from amino acid oxidative metabolism, acting in conjunction with glucose-signalled messengers, are very effective at closing ATP-dependent K+ channels (KATP+). 8. We propose that metabolic regulation of KATP+ channels is one of the mechanisms underlying the modulation of the oscillatory [Ca2+]i response to nutrient secretagogues.

Amino Acids↗

PubMed1995

Ubiquitinylation is not an absolute requirement for degradation of c-Jun protein by the 26 S proteasome.

Degradation of rapidly turned over cellular proteins is commonly thought to be energy dependent, to require tagging of protein substrates by multi-ubiquitin chains, and to involve the 26 S proteasome, which is the major neutral proteolytic activity in both the cytosol and the nucleus. The c-Jun oncoprotein is very unstable in vivo. Using cell-free degradation assays, we show that ubiquitinylation, along with other types of tagging, is not an absolute prerequisite for ATP-dependent degradation of c-Jun by the 26 S proteasome. This indicates that a protein may bear intrinsic structural determinants allowing its selective recognition and breakdown by the 26 S proteasome. Moreover, taken together with observations by different groups, our data point to the notion of the existence of multiple degradation pathways operating on c-Jun.

Animals↗

PubMed1995

Identification and characterization of a soluble c-kit receptor produced by human hematopoietic cell lines.

Stem cell factor (SCF) triggers cell growth by binding to cell surface c-kit receptors. Soluble forms of several cytokine receptors have been described and may play a role in the modulation of cytokine activity in vivo. For these reasons, we investigated whether human hematopoietic cells produce soluble c-kit receptors. The human leukemia cell lines OCIM1 and MO7e display approximately 80,000 and approximately 35,000 high-affinity cell surface c-kit receptors, respectively. Soluble c-kit receptors were detected by enzyme immunoassay in OCIM1 and MO7e culture supernatants. We determined the molecular weight and binding affinity of soluble c-kit receptor produced by OCIM1 cells, soluble c-kit receptor purified from human serum, and recombinant soluble c-kit receptor expressed in CHO cells. The three soluble c-kit receptors each have a molecular weight of 98 kD. Quantitative binding experiments with 125I-SCF indicate that the soluble c-kit receptors obtained from human serum or OCIM1 cells have binding affinities for SCF of approximately 200 to 300 pmol/L, in contrast to the recombinant form, which has a binding affinity of approximately 1.5 nmol/L. All three forms of the soluble c-kit receptor were able to compete with c-kit receptors on OCIM1 cells for 125I-SCF binding. Thus human hematopoietic cells can produce a soluble form of the c-kit receptor that retains high-affinity SCF binding activity. We speculate that the soluble c-kit receptor may bind SCF and function as a receptor antagonist in vivo.

Amino Acid Sequence↗

PubMed1995

Genetic selection for active E.coli amber tRNA(Asn) exclusively led to glutamine inserting suppressors.

Suppressor tRNAs are useful tools for determining identity elements which define recognition of tRNAs in vivo by their cognate aminoacyl-tRNA synthetases. This study was aimed at the isolation of active amber tRNA(Asn). Nineteen mutated tRNA(Asn)CUA having amber suppressor activity were selected by an in vivo genetic screen, and all exclusively inserted glutamine. From analysis of the different mutations it is concluded that glutamine accepting activity was obtained upon reducing the interaction strength between the first base pair of the tRNA(Asn)CUA by direct or indirect effects. Failure to isolate tRNA(Asn)CUA suppressors charged with asparagine as well as other evolutionary related amino acids is discussed.

Base Sequence↗