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C Martinez

Publications and source records attributed to C Martinez.

At least 19 recordsLinked to original sources
PubMed1995

Impairment of the liver insulin receptor autoactivation cascade at full-term pregnancy in the rat.

Partially purified liver insulin receptors from full-term pregnant rats show decreased autophosphorylation rates if compared with receptors from virgins. We studied the molecular mechanism of this phenomenon, looking at possible structural and functional changes of several domains. The ATP-binding domain seems to be unaltered in receptors from pregnant rats since Km for ATP was similar to that observed in virgins. In contrast, the Vmax. is decreased some 45%, suggesting changes in the kinase domain. Truncation of a fragment of 10 kDa from the C-terminal tail does not normalize the kinase activity in receptors from pregnant rats, suggesting that this domain is not involved in the inhibitory regulation. Treatment with alkaline phosphatase increases the [32P]Pi incorporation into receptors from pregnant rats; however, the autophosphorylation remains lower than that observed in virgin rats. Tryptic phosphopeptide maps of phosphorylated receptors show that the same phosphopeptides are present in receptors from virgin and pregnant rats. However, the progression through the autoactivation cascade in the kinase domain is impaired in receptors from pregnant rats. Differences in the cleavage by trypsin at the two alternative sites in the kinase domain were observed, indicating possible structural changes in receptors from pregnant rats that could be related to the impairment of the autoactivation cascade. Integrity of the alpha- and beta-subunits, as well as differential expression of the two receptor isotypes, were shown to be unaltered. We conclude that (1) the decreased autophosphorylation rate of the liver insulin receptor from pregnant rats is associated with the impairment of its autoactivation cascade, probably as a consequence of the basal Ser/Thr phosphorylation; and (2) the inhibition of the autoactivation cascade does not account for the overall inhibition of autophosphorylation observed in receptors from pregnant rats.

Adenosine Triphosphate↗
PubMed1995

Role of the autophosphorylation site on the biological function of pp56lck.

Src-family tyrosine kinases act as signaling molecules in a wide array of cellular activation processes. The existence of the various src-family members reflects the requirement for different cell-surface receptors to transmit cell-type specific intracellular signals. The structural basis for the functional specificity of src-kinases is being actively investigated. In the present report we have analysed the contribution of the area surrounding the autophosphorylation site (located at subdomain VII of the catalytic domain) in determining src-kinases activity and functional specificity. To this end we analysed the kinase activities of the lymphoid src-kinase pp56lck and a mutant of pp56lck in which this region has been exchanged for the corresponding area of the serine/threonine kinase c-Raf. Our studies indicate that the change at subdomain VII affected the ability of pp56lck to phosphorylate physiological substrates. Furthermore, when analysed in T cells, the mutant at subdomain VII failed to induce interleukin-2 production, a specific biological function of pp56lck. Thus, the area surrounding the autophosphorylation site of pp56lck plays a critical role in mediating its specific biological function.

Base Sequence↗
PubMed1995

Lck unique domain influences Lck specificity and biological function.

Src-family tyrosine kinases share structural and amino acid sequence homology, particularly in the catalytic domain as well as in the SH2 and SH3 domains of the regulatory region. However, each src-family member also contains a unique domain which is specific to and characteristic of each individual tyrosine kinase. These unique or specific domains may contribute to the functional specificity of each src-family kinase. To address this possibility, we analyzed the kinase activities and substrate specificities of the lymphoid src-kinase, pp56lck, and a mutant of pp56lck lacking its specific domain. Our data show that both the wild type enzyme and the specific domain-deleted mutant displayed similar affinities for ATP and the non-physiological substrate denatured enolase. However, the specific domain-deleted mutant failed to phosphorylate a number of physiological substrates of pp56lck. In addition, the ability of pp56lck to mediate induction of the interleukin-2 promoter was strongly impaired upon deletion of its specific domain. Thus, the unique domain is not required for the intrinsic kinase activity of pp56lck, however, it influences substrate preference and contributes to the unique physiological function of this src-family tyrosine kinase.

Base Sequence↗
PubMed1995

Balloon-expandable stent repair of severe coarctation of aorta.

Experimental studies have shown that stents implanted at the aorta become incorporated within the aortic wall and can be further expanded in growing animals. This study evaluates the feasibility and immediate results of balloon-expandable stent implantation in 10 patients with severe coarctation of aorta. The ages of the patients ranged from 1 month to 43 years; 1 was an infant, 8 were children (mean age 5.3 +/- 4 years), and 1 was an adult. All had an unfavorable anatomy for balloon angioplasty; 9 had isthmus hypoplasia. Balloon predilation was first performed and its immediate effect evaluated. Then a balloon-expandable stent that was 30 mm long and covered the isthmus and coarctation levels was deployed, and it was further expanded to the preselected final diameter (12 +/- 4 mm). A final hemodynamic and angiographic evaluation was then obtained. Full deployment of an incompletely expanded and distally displaced stent in the infant led to aortic disruption that was controlled by a second stent covering the disrupted zone and the isthmus. After balloon angioplasty alone was done, the mean gradient (43 +/- 12 vs 31 +/- 10 mm Hg) and the percentage stenosis (72% +/- 11% vs 54% +/- 11%) had an insufficient decrease. However, after stent implantation was done, the gradient almost disappeared (mean 2 +/- 3 mm Hg). The angiographic stenosis disappeared in 7 patients and was markedly reduced in 3. The ratio of isthmus/descending aorta changed from 0.65 +/- 0.14 to 1 +/- 0.08 (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Disease↗
PubMed1995

Risk factors for acquisition of urinary tract infections caused by ciprofloxacin resistant Escherichia coli.

From January 1990 to December 1992, ciprofloxacin resistant Escherichia coli was isolated in 125 of 1,946 urine cultures (6.4%) with more than 10(5) colony-forming units per ml. To determine the risk factors for acquisition of urinary tract infections caused by ciprofloxacin resistant E. coli a retrospective chart review was done. Data from 54 patients with urinary tract infections caused by ciprofloxacin resistant E. coli were compared with 51 controls matched by temporal occurrence and randomly selected among 540 patients with urinary tract infections caused by ciprofloxacin susceptible E. coli. Patients had greater proportions of asymptomatic bacteriuria or lower urinary tract symptoms (85% versus 61%, p = 0.01) and of relapse (22% versus 0%, p = 0.001) than controls. Urinary tract abnormalities (odds ratio 7.98, 95% confidence interval 2.7 to 3.1, p < 0.001), patient age 65 years or older (odds ratio 6.48, 95% confidence interval 2.2 to 19.1, p < 0.001), previous treatment with quinolones (odds ratio 19.09, 95% confidence interval 2.2 to 166.5, p = 0.008) and urinary catheterization (odds ratio 2.92, 95% confidence interval 1.1 to 8.5, p = 0.048) were independently associated with infections caused by ciprofloxacin resistant strains. Our results suggest that patients with urological abnormalities previously treated with quinolones are especially prone to urinary tract infection caused by ciprofloxacin resistant strains.

Age Factors↗
PubMed1995

Energy expenditure and body fat distribution in Mohawk children.

OBJECTIVE: Epidemiologic studies suggest that Native Americans, including the Mohawk people, have a high prevalence of obesity, diabetes, and cardiovascular risk. However, current information on alterations in related variables such as energy metabolism and body composition in Native Americans is almost exclusively limited to already obese Pima adults living in the Southwest. The aim of this study was to characterize energy metabolism and body composition in young Mohawk children (17 girls, 11 boys; aged 4 to 7 years) as compared to Caucasian children (36 girls, 34 boys; aged 4 to 7 years). Total energy expenditure was measured by doubly labeled water, postprandial resting energy expenditure by indirect calorimetry, and activity energy expenditure was derived from the difference between total and resting energy expenditure. Fat and fat free mass were estimated from bioelectrical resistance, and body fat distribution was estimated from skinfolds and circumferences. RESULTS: There were no significant effects of ethnic background or sex on body weight, height, or body mass index. Fat free mass was significantly higher in boys and fat mass was significantly higher in girls, with no effect of ethnic background. Chest skinfold thickness, the ratio of trunk skinfolds:extremity skinfolds, and the waist:hip ratio were significantly higher in Mohawk children by 2.5 mm, 0.09 units, and 0.03 units, respectively, independent of sex and fat mass. Total energy expenditure was significantly higher in Mohawk children compared to Caucasian (100 kcal/day in girls, 150 kcal/day in boys), independent of fat free mass and sex, due to a significantly higher physical activity-related energy expenditure. CONCLUSION: These data suggest that: 1) body fat is more centrally distributed in Mohawk relative to Caucasian children, and this effect is independent of sex and body fat content; 2) Mohawk children have a greater total energy expenditure than Caucasian children, independent of fat free mass, due to greater physical activity-related energy expenditure.

Adipose Tissue↗
PubMed1994

Tyrosine kinase specific motif at subdomain VIII does not confer specificity for tyrosine.

The majority of protein kinases fall within one of the two broad classes, kinases that phosphorylate serine or threonine and kinases that phosphorylate tyrosine. The structural basis that confers residue specificity is not known. However, it has been hypothesized that a region in subdomain VIII of the catalytic domain may be involved in determining kinase specificity. This region contains a motif which is conserved among serine/threonine kinases and different from the one conserved among tyrosine kinases. We have prepared a chimera of the tyrosine kinase pp56lck in which the tyrosine kinase motif at subdomain VIII has been exchanged for the corresponding region of the serine/threonine kinase c-Raf. Our results indicate that this motif itself does not confer amino acid specificity since the chimeric kinase still displays specificity for tyrosine.

Amino Acid Sequence↗
PubMed1994

Gene expression of VIP receptor in rat lymphocytes.

In the present report we analyze by reverse transcription (RT) and polymerase chain reaction (PCR) the expression of vasoactive intestinal peptide receptors (VIP-R) in rat thymocytes and peripheral T and B lymphocytes. We have obtained a specific VIP-R cDNA product of 311 bp identical in size to that obtained from lung. These results have been confirmed by Southern hybridization analysis. VIP receptor gene expression in central and peripheral lymphoid organs support the notion that VIP is a immunomodulatory peptide.

Adjuvants, Immunologic↗
PubMed1994

Horse pancreatic lipase. The crystal structure refined at 2.3 A resolution.

Pancreatic lipase (EC 3.1.1.3) plays a key role in dietary fat digestion by converting triacylglycerols into 2-monoacylglycerols and free fatty acids in the intestine. Although the crystallographic structures of the human pancreatic lipase and of a human lipase-porcine colipase complex have been solved, no refined structure of pancreatic lipase has yet been published. The crystal structure of the horse enzyme was solved by the molecular replacement method from the model of the human pancreatic lipase and subsequently refined to 2.3 A resolution. The final model contains two molecules of 449 amino acid residues each in the asymmetric unit, 705 well-defined water molecules and two calcium ions. The two molecules in the asymmetric unit of the orthorhombic crystals are related by a 2-fold non-crystallographic symmetry axis as in the case of the human lipase. However, the association between the two molecules in their respective crystal forms is different. The overall molecular structure of the horse lipase is very similar to that of the human enzyme. The horse lipase is made up of two well-defined domains. The N-terminal domain which bears the active centre has a typical alpha/beta hydrolase fold topology. The C-terminal domain which is devoted to colipase binding has a beta-sheet sandwich topology. Comparison of equivalent C alpha atom positions between the final model of the horse lipase and the human lipase structure shows only slight differences which are mainly located in the C-terminal domain. The horse enzyme possesses the common features of the known mammalian and microbial lipases, in particular the "flap" covering the catalytic triad. In addition to more precise information concerning these features, the elucidation of the horse lipase crystal structure allowed us to better understand the structural basis of the kinetic behaviour of pancreatic lipases towards a soluble substrate, p-nitrophenyl acetate, and the different sensitivity of these enzymes towards limited proteolysis.

Amino Acid Sequence↗
PubMed1994

Repeated stress causes reversible impairments of spatial memory performance.

Restraint stress, 6 h/day for 21 days, caused an impairment, during acquisition, of the performance of a spatial memory task, the eight-arm radial maze. The impairment was reversible, temporally limited and blocked by phenytoin, a blocker of excitatory amino acid action, or tianeptine, an antidepressant, which lowers extracellular serotonin. These effects on behavior parallel the reversible stress-induced atrophy of dendrites of hippocampal CA3 neurons that are also blocked by the drugs.

Animals↗
PubMed1994

Cutinase, a lipolytic enzyme with a preformed oxyanion hole.

Cutinases, a group of cutin degrading enzymes with molecular masses of around 22-25 kDa (Kolattukudy, 1984), are also able to efficiently hydrolyse triglycerides (De Geus et al., 1989; Lauwereys et al., 1991), but without exhibiting the interfacial activation phenomenom (Sarda et al., 1958). They belong to a class of proteins with a common structural framework, called the alpha/beta hydrolase fold (Martinez et al., 1992; Ollis et al., 1992). We describe herein the structure of cutinase covalently inhibited by diethyl-p-nitrophenyl phosphate (E600) and refined at 1.9-A resolution. Contrary to what has previously been reported with lipases (Brzozowski et al., 1991; Derewenda et al., 1992; Van Tilbeurgh et al., 1993), no significant structural rearrangement was observed here in cutinase upon the inhibitor binding. Moreover, the structure of the active site machinery, consisting of a catalytic triad (S120, H188, D175) and an oxyanion hole (Q121 and S42), was found to be identical to that of the native enzyme, whereas the oxyanion hole of Rhizomucor lipase (Brzozowski et al., 1991; Derewenda et al., 1992), like that of pancreatic lipase (van Tilbeurgh et al., 1993), is formed only upon enzyme-ligand complex formation. The fact that cutinase does not display interfacial activation cannot therefore only be due to the absence of a lid but might also be attributable to the presence of a preformed oxyanion hole.

Amino Acid Sequence↗
PubMed1994

Manganese intoxication and chronic liver failure.

Manganese intoxication and chronic liver failure are associated with strikingly similar clinical, imaging, and pathological abnormalities. As manganese is primarily cleared by the liver, inadequate elimination of manganese absorbed from the normal diet may lead to manganese overload in patients with liver disease. We report a significant elevation of blood manganese concentration in 3 patients with biopsy-proved hepatic cirrhosis who exhibited neurological dysfunction and characteristic abnormal signal hyperintensity in the globi pallidi and substantia nigra on T1-weighted magnetic resonance imaging. We speculate that manganese accumulation in the brain accounts for the magnetic resonance imaging abnormalities and contributes to neurological dysfunction in patients with liver disease.

Aged↗
PubMed1994

CD2-CD48 interaction prevents apoptosis in murine B lymphocytes by up-regulating bcl-2 expression.

Antigen receptor engagement initiates clonal expansion and antibody secretion in B lymphocytes in response to foreign antigens. However, binding of self antigen to antigen receptors targets self-reactive B cell clones for elimination or inactivation. The antigen-triggered biochemical events and the eventual response of the cells are dependent on the simultaneous occupancy of co-stimulatory receptors. CD2 is an intercellular adhesion molecule implicated in cell activation and expressed in human T and natural killer cells as well as in mouse B lymphocytes. Mouse B cells specific for allogeneic major histocompatibility complex (MHC) class I initiate a suicide program that leads to DNA fragmentation and cell death when confronted with soluble MHC class I while undergoing clonal expansion when the antigen is present on mitomycin C-treated cells. Here we show that occupancy of CD2 in mouse B cells by the presence of either monoclonal antibody (mAb) specific for CD2, or soluble recombinant mouse CD48, its natural ligand in mouse, prevents the induction of apoptosis. Furthermore, the in vitro activation by mitomycin C-treated allogeneic cells, is abrogated in the presence of anti-CD48 mAb (OX78). These results indicate that a CD2-CD48 interaction is involved in the control of B cell activation.

Animals↗
PubMed1994

Activity of peptidergic neurons in the amygdala during sexual behavior in the male rat.

The medial amygdaloid nucleus (AME) occupies a central position in the circuitry that organizes sexual behavior in the male rat. It receives a projection from olfactory structures that are activated by pheromonal cues indicating receptivity in the female and projects in turn to limbic and hypothalamic structures that are thought to organize aspects of coitus. Electrical stimulation of the AME elicits a behavioral state that is indistinguishable by several measures from the post-ejaculatory interval. We used chronic single-unit recording techniques to determine the behavioral conditions in which the AME is normally active. We found that the cells indeed fired selectively during the presence of a receptive female, but that the discharge considerably anticipated copulation in time. We propose that sexual behavior in the male rat is a reaction chain of fixed action patterns, each one acting as a releaser for the next. The AME mediates an early event in the reaction chain, namely recognition of the receptive female, but electrical activation of the AME causes the reaction chain to proceed to its culminating behavior, the post-ejaculatory interval.

Amygdala↗