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Biomedical subjects

M Torres

Publications and source records attributed to M Torres.

At least 37 records · Page 2Linked to original sources
PubMed1991

Paradoxical potentiation by low extracellular Ca2+ of acute chemical anoxic neuronal injury in cerebellar granule cell culture.

Acute chemical anoxic injury was produced in primary cerebellar granule cell cultures incubated with iodoacetate (IAA) alone or IAA combined with potassium cyanide (KCN). Cytotoxicity was assessed using Trypan blue exclusion or LDH release. Four millimolars of KCN induced approx 30% neuron death at 3 h, whereas greater than 50% cell death was produced by 0.2 mM IAA. No potentiation of cytotoxicity was observed by IAA + KCN. A total of 0.2 mM IAA produced an early major reduction of intracellular ATP prior to the onset of neuron injury or reduction in intracellular glutathione (GSH). Medium Na+ replacement by choline, K+, or methylglucamine protected against IAA-induced neuronal injury, reduced the rate of decline of intracellular ATP but had no effect on intracellular GSH. Some 80% neuronal survival was obtained when Na+ was deleted from the medium even after the intracellular ATP had been reduced to less than 10% of control. Removal of Ca2+ from the medium had no effect on control culture, Trypan blue exclusion, GSH, or ATP, but potentiated the onset and magnitude of IAA-induced cytotoxicity. ATP and GSH decline. Loading of granule cells with the Ca2+ chelator Fura-2 did not influence IAA-induced cytotoxicity in control or low Ca2+ media. Addition of 50 microM glutamate had a minimal cytotoxic effect over 3 h and the combined addition of 0.2 mM IAA plus 50 microM glutamate did not potentiate IAA-induced injury. The glutamate receptor antagonists, D-2-amino-5-phosphonovaleric acid (APV) or kynurenate did not block IAA-induced injury in control medium but inhibited the potentiation of toxicity seen in the low Ca2+ medium. This study suggests the use of IAA as a chemical anoxic agent in cerebellar granule cell culture. The early, dose-dependent decline in ATP may be dissociated from GSH change. Acute IAA-induced injury is Na+/Cl- dependent but paradoxically potentiated in low Ca2+ medium. The low Ca2+ potentiated component was sensitive to glutamate/NMDA receptor antagonists and associated with reduction of intracellular GSH.

2-Amino-5-phosphonovalerate↗
PubMed1991

Carbachol induced release of diadenosine polyphosphates--Ap4A and Ap5A--from perfused bovine adrenal medulla and isolated chromaffin cells.

The diadenosine polyphosphates--Ap4A and Ap5A--were released from perfused bovine adrenal glands and recently isolated chromaffin cells by the action of carbachol. The H.P.L.C. technique reported here allowed the quantification of pmol amounts of these compounds present in biological samples from the perfusion media after stimulation. Both compounds (Ap4A and Ap5A) were identified by the retention time in H.P.L.C. chromatography, co-elution with standards, re-chromatography and destruction by the phosphodiesterase action. Bovine adrenal glands stimulated with 100 microM carbachol released 0.47 +/- 0.12 nmol/gland of Ap4A and 1.11 +/- 0.26 nmol/gland of Ap5A. Isolated bovine chromaffin cells after 100 microM carbachol, as secretagogue, released 11.1 +/- 0.8 pmol/10(6) cells of Ap4A and 15.8 +/- 1.1 pmol/10(6) cells of Ap5A. The ratio of these compounds with respect to the exocytotically released ATP and catecholamines was in the same order as that found in isolated chromaffin granules.

Adenosine Triphosphate↗
PubMed1991

Further evidence for a dopaminergic involvement in the renal action of centrally administered atrial natriuretic peptide in rats.

Intracerebroventricular (ICV) administration of atrial natriuretic peptide (ANP) induces an increase of urinary volume and sodium excretion in conscious male hydrated rats. The involvement of brain dopaminergic neurones in the ICV-ANP renal action was investigated. Diuretic and natriuretic action of ANP was prevented by inhibition of tyrosine hydroxylase activity with alpha-methyl-p-tyrosine treatment and was absent following selective central dopaminergic denervation with ICV administration of 6-hydroxydopamine in combination with desmethylimipramine (IP). Our results suggest that ANP acts centrally, at least in part, via an interaction with endogenous dopamine neurones.

Analysis of Variance↗
PubMed1991

Prevalence of HIV antibody in a noninner-city university hospital emergency department.

STUDY HYPOTHESIS: To determine the prevalence of antibody to human immunodeficiency virus (HIV) in trauma and nontrauma patients not identified as having known HIV infection in a noninner-city university teaching hospital emergency department, and to determine the frequency with which treating emergency physicians are knowledgeable of patients' risk factors for HIV infection. POPULATION: ED patients between 18 and 59 years old with injuries that met trauma center triage criteria or with nontrauma-related illness who had blood drawn for physician-requested laboratory tests and for whom an extra aliquot of blood was available for HIV antibody testing. METHODS: All serum samples were first tested for antibody to HIV by enzyme-linked immunosorbent assay. If positive, the specimen was retested. All repeatedly reactive specimens were analyzed by Western blot test. The treating physician completed a questionnaire regarding the patient's illness and risk factors for HIV infection after the patient's care was completed. RESULTS: Two of 100 major trauma patients (confidence interval, 0% to 5%) and seven of 100 nontrauma patients (confidence interval, 2% to 12%) had antibody to HIV. The seropositive rate by age and clinical group varied from 0% to 12.5%, with the highest rates in the 30- to 39-year-old group of nontrauma patients. The difference in proportions of seropositivity between the sexes was not statistically significant. Physicians obtained information regarding homosexual or bisexual behavior, IV drug use, and hemophilia from 52% of the nontrauma patients and only 17% of trauma patients. None of 30 trauma patients for whom data were available and only two of the 100 nontrauma patients gave a history of any high-risk behavior. CONCLUSION: Although the sampling technique we used has limitations, the prevalence of HIV infection in our noninner-city ED is similar to that recently reported from inner-city EDs. This is in contrast to previous reports of low rates of HIV infection among ED patients in nonurban settings. Physician assessment of risk factors was incomplete in the majority of our patients. Patients rarely acknowledged any high-risk behavior. It is essential that emergency health care workers take maximum diligence to prevent exposure to blood and other body fluids from all ED patients.

Adult↗
PubMed1991

Genetic and molecular analysis of new female-specific lethal mutations at the gene Sxl of Drosophila melanogaster.

We have isolated three female-specific lethal mutations at the gene Sex-lethal (Sxl): Sxlfb, Sxlfc and Sxlfd. We have carried out the complementation analysis between these mutations and other previously reported Sxlf mutations. It is possible to classify the alleles tested in this report into two complementation groups: the bc group defined by Sxlfb, and Sxlfc, and the LS group defined by SxlfLS. The other alleles tested affect both complementation groups albeit with different degrees. Contrary to what happens with mutations at the LS group, mutations at the bc group do not affect sex determination, nor late dosage compensation nor oogenesis. Both Sxlfb and Sxlfc present a DNA insertion of at least 5 kb between position -10 and -11 on the molecular map, within the fourth intron. On the contrary, Sxlfd, a strong mutation affecting all Sxl functions, is not associated to any detectable DNA alteration in Southern blots, so that it seems to be a "point" mutation. In agreement with their phenotypes, both Sxlfc/SxlfLS and Sxlfc homozygous female larvae express only the late Sxl transcripts characteristic of females, while females homozygous for SxlfLS express only the late Sxl transcripts characteristic of males. Moreover, Sxlfc presents a lethal synergistic interaction with mutations at either da or the X:A ratio, two signals that define the initial activity state of Sxl, while SxlfLS do not. These data suggest that the two complementation groups are related to the two sets of early and late Sxl transcripts, which are responsible for the early and late Sxl functions, respectively: Sxlfb and Sxlfc would affect the early functions and SxlfLS would affect the late Sxl functions.

Alleles↗
PubMed1991

Immunoscintigraphy of lung cancer using 111In-labelled antiCEA F023C5-F(ab')2 fragments.

Forty-one patients with known lung carcinoma or suspicion of bearing this disease were studied with 111In-antiCEA F023C5-F(ab')2. Five patients with positive results were further studied to assess in vivo specificity using 111In-4C4-F(ab')2 (MoAb antihepatitis). Immunoscintigraphic results have been compared to immunohistochemistry in 16 patients. Tumour visualization with non-specific MoAb was present in all five patients although the maximum tumour/background (T/B) ratio obtained was 1.51. Therefore a 'specificity criterion' has been applied to examinations performed with MoAb antiCEA. There was considered to be a positive result when the T/B ratio was greater than 1.60. Results obtained in this way for thoracic lesions were 25/34 true positive (TP) (73%) and 6/7 true negative (TN) (86%). When studies were analysed visually only, the results were 32/34 TP (94%) and 3/7 TN (43%). In addition, immunoscintigraphic results are more concordant with immunohistochemical results when the specificity criterion is applied. In metastatic lesions results were 9/13 TP (69%) with only 1/3 TP (33%) for liver metastasis (anatomic site of lowest detection ability). SPECT imaging did not improve results over planar images, although it clearly contributed to a better anatomic location of lesions. The constant presence of non-specific uptake in thoracic lesions makes it necessary to establish a T/B ratio limit in order to obtain adequate specificity with the immunoscintigraphic technique.

Adenocarcinoma↗
PubMed1991

Ca(2+)-independent release of glutamate during in vitro anoxia in isolated nerve terminals.

The effects of in vitro anoxia on the release of glutamate in isolated nerve terminals were studied. The extra-synaptosomal concentration of glutamate ([Glu]ext) under aerobic conditions was 2.3 microM and increased to 4.9 microM after 10 min of anoxia. However, when synaptosomes were incubated in the presence of lactate plus pyruvate instead of glucose, to prevent anaerobic glycolysis, anoxia induced an eightfold increase in the [Glu]ext. The accumulation of glutamate in the external medium during anoxia was Ca2+ independent and insensitive to a significant reduction of the Ca(2+)-dependent release of the amino acid. These results indicate that a Ca(2+)-independent efflux of cytoplasmic glutamate occurs during in vitro anoxia in isolated nerve terminals.

4-Aminopyridine↗
PubMed1991

Characterization of diadenosine tetraphosphate (Ap4A) binding sites in cultured chromaffin cells: evidence for a P2y site.

1. Diadenosine tetraphosphate (Ap4A) a dinucleotide, which is stored in secretory granules, presents two types of high affinity binding sites in chromaffin cells. A Kd value of 8 +/- 0.65 x 10(-11) M and Bmax value of 5420 +/- 450 sites per cell were obtained for the high affinity binding site. A Kd value of 5.6 +/- 0.53 x 10(-9) M and a Bmax value close to 70,000 sites per cell were obtained for the second binding site with high affinity. 2. The diadenosine polyphosphates, Ap3A, Ap4A, Ap5A and Ap6A, displaced [3H]-Ap4A from the two binding sites, the Ki values being 1.0 nM, 0.013 nM, 0.013 nM and 0.013 nM for the very high affinity binding site and 0.5 microM, 0.13 microM, 0.062 microM and 0.75 microM for the second binding site. 3. The ATP analogues displaced [3H]-Ap4A with the potency order of the P2y receptors, adenosine 5'-O-(2 thiodiphosphate) (ADP-beta-S) greater than 5'-adenylyl imidodiphosphate (AMP-PNP) greater than alpha, beta-methylene ATP (alpha, beta-MeATP), in both binding sites. The Ki values were respectively 0.075 nM, 0.2 nM and 0.75 nM for the very high affinity binding site and 0.125 microM, 0.5 microM and 0.9 microM for the second binding site.

Animals↗
PubMed1991

The sisterless-b function of the Drosophila gene scute is restricted to the stage when the X:A ratio determines the activity of Sex-lethal.

The gene scute (sc) has a dual function: the scute function which is involved in neurogenesis and the sisterless-b function which is involved in generating the X:A signal that determines the state of activity of Sxl, a gene that controls sex determination and dosage compensation. We show here that the lethal phase of sc- females is embryonic and caused by the lack of Sxl function. We also analyze the time in development when sc and Sxl interact by means of (a) determining the thermosensitive phase (TSP) of the interaction between Sxl and sc and (b) a chimeric gene in which sc is under the control of a heat-shock promoter (HSSC-3). Pulses of sc expression from the HSSC-3 activate Sxl only at a very specific and early stage in development, which coincides with the TSP of the interaction between sc and Sxl. It corresponds to the syncytial blastoderm stage and coincides with the time when the X:A signal regulates Sxl. At this stage sc undergoes a homogeneous transient expression in wild-type flies. We conclude that the sc expression at the syncytial blastoderm is responsible for its sisterless-b function. Since sc expression from the HSSC-3 fully suppresses the sisterless-b phenotype, we further conclude that the sisterless-b function is exclusively provided by the sc protein. Finally, we have analyzed, by in situ hybridization, the effect of sc and sis-a mutations on the embryonic transcription of Sxl. Our results support the view that the control of Sxl by the X:A signal occurs at the transcriptional level.

Animals↗
PubMed1990

Presence of ectonucleotidases in cultured chromaffin cells: hydrolysis of extracellular adenine nucleotides.

The granular ATP released from chromaffin cells during the secretory response can be hydrolyzed by ectonucleotidases that are present in the plasma membrane of these cells. The ecto-ATPase activity showed a Km for ATP of 250 +/- 18 microM and a VMAX value of 167 +/- 25 nmol/10(6) cells x min (1.67 mumol/mg protein x min) for cultured chromaffin cells, while the ecto-ADPase activity showed a Km value for ADP of 375 +/- 40 microM and a VMAX of 125 +/- 20 nmol/10(6) cells x min (1.25 mumol/mg protein x min). The ecto 5'-nucleotidase activity of cultured chromaffin cells was more specific for the purine nucleotides, AMP and IMP, than for the pirimidine nucleotides, CMP and TMP. The Km for AMP was 55 +/- 5 microM and the VMAX value was 4.3 +/- 0.8 nmol/10(6) cells x min (43 nmol/mg protein x min). The nonhydrolyzable analogs of ADP and ATP, alpha, beta-methylene-adenosine 5'-diphosphate and adenylyl-(beta, gamma-methylene)-diphosphonate were good inhibitors of ecto 5'-nucleotidase activity, the KI values being 73.3 +/- 3.5 nM and 193 +/- 29 nM, respectively. The phosphatidylinositol-specific phospholipase C released the ecto-5'-nucleotidase from the chromaffin cells in culture, thus suggesting an anchorage through phosphatidylinositol to plasma membranes. The presence of ectonucleotidases in chromaffin cells may permit the recycling of the extracellular ATP exocytotically released from these neural cells.

Adenine Nucleotides↗
PubMed1990

All nucleoside transporters in bovine chromaffin cells are nitrobenzylthioinosine sensitive.

Nitrobenzylthioinosine is an effective inhibitor of adenosine transport in chromaffin cells. When adenosine transport was measured at a 0.15 microM adenosine concentration, in the presence of variable concentrations of nitrobenzylthioinosine, ranging from 10(-14) to 10(-6) M, a half-maximal inhibitory concentration value (IC50) of 1 +/- 0.3 nM was deduced. This compound has the capacity to inhibit the total adenosine transport at 10(-7) M concentration. Nitrobenzylthioinosine acts in a non-competitive manner in blocking adenosine transport, as deduced from a Dixon plot, with a constant inhibition value (K1) of 0.01 +/- 0.003 nM. The results suggest that all nucleoside transporters present in bovine chromaffin cells are sensitive to the nitrobenzylthioinosine inhibition.

Adenosine↗
PubMed1990

Use of a piezoelectric film sensor for monitoring vascular grafts.

Detection of failing arterial reconstructions requires intensive surveillance by frequent physical examination and noninvasive laboratory testing. However, many grafts fail during the intervals between these examinations. For this reason, we have developed an implantable miniaturized piezoelectric flow detection device whose function can be monitored externally by radiotransmission across the skin. Sensors were constructed from ultrathin polyvinylidene fluoride (PVF2) with piezoelectric activity and attached with silicone fixative to 6-mm polytetrafluoroethylene grafts. Ten of these grafts were placed in mongrel dogs as iliofemoral bypasses. Real time data were acquired from the sensors at a rate of 200 Hz, using a DATAQ A/D data acquisition board and CODAS data acquisition software, while simultaneous blood flow (using an electromagnetic flowmeter) and intraluminal pressure were processed by using separate channels of the same data acquisition board. The data were stored on computer storage media and analyzed by the ASYST software, which allows simultaneous signal curves to be compared using regression analysis. In the resting state, the mean blood flow was 123 +/- 16 mL and the mean intraluminal pressure was 124/78 mm Hg, and there was perfect correlation between the PVF2 sensor and the flowmeter and between the sensor and the intraluminal pressure (correlation coefficient, r greater than or equal to 0.99 and r greater than or equal to 0.93, respectively). A tourniquet was applied to the iliac artery proximal to the graft to reduce the flow to approximately half of the resting state (mean flow after tourniquet: 66 +/- 6 mL/minute). Signal tracings from the three sources showed a remarkable similarity with a very high correlation coefficient (r greater than or equal to 0.99 between sensor and flowmeter and r greater than or equal to 0.92 between sensor and the pressure signal). These preliminary results show that the sensors made from low-profile and low-mass PVF2 material have the potential of being implanted around grafts for long-term, continuous monitoring of graft function. Further studies involving long-term implantation to assess the effect of tissue ingrowth and loss of compliance are necessary before this device can be used clinically.

Animals↗
PubMed1990

Multiple endocrine neoplasia type II B with symptoms suggesting Hirschsprung's disease: a case report.

A 3-year-old child was referred with a tentative diagnosis of Hirschsprung's disease because of life-long constipation and "megacolon" demonstrated radiographically. Our rectal biopsy revealed hyperganglionosis suggestive of multiple endocrine neoplasia (MEN) type II B. This, in addition to an elevated serum calcitonin level, prompted surgical removal of her thyroid, which appeared grossly normal but on sectioning, contained a medullary carcinoma in each lobe. She remains disease-free 5 years later. Gastrointestinal symptoms are a significant component of the MEN type II B syndrome, and often antedate the full phenotypic expression of the syndrome and the development of potentially lethal endocrine neoplasms. On the basis of this experience, it is recommended that MEN II B be included in the differential diagnosis of chronic constipation.

Child, Preschool↗