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G Vazquez

Publications and source records attributed to G Vazquez.

At least 37 records · Page 2Linked to original sources
PubMed1999

Activation of a beta-adrenergic-sensitive signal transduction pathway by the secosteroid hormone 1,25-(OH)2-vitamin D3 in chick heart.

In recent studies we have established that 1 alpha, 25-dihydroxy-vitamin D3[1,25(OH)2D3] rapidly stimulates dihydropyridine-sensitive calcium channel-mediated Ca2+influx in chick cardiac muscle by a non-genomic action which is accompanied by PKA-dependent phosphorylation of a 45 kDa microsomal membrane protein. To investigate the signal transduction pathway activated by 1,25(OH)2D3 in heart, we have compared the effects of the secosteroid hormone with those of the beta-adrenergic agonist isoproterenol (IPT) by employing cultured chick embryonic cardiac cells (myocytes) and thin-slice preparations of differentiated adult heart muscle. The increases in 45Ca2+ uptake and intracellular calcium ([Ca2+]i), cyclic AMP accumulation and changes in microsomal protein phosphorylation evoked by 1,25(OH)2D3 could be reproduced by IPT. When combined treatments with the sterol and the beta-adrenergic agonist were performed, no additive stimulation of these parameters was observed, suggesting that a common signal transduction pathway mediates the effects of 1,25(OH)2D3 and IPT. The participation of a guanine nucleotide binding protein (G protein) in the 1, 25(OH)2D3-induced changes in heart was investigated. AlF4(-), an activator of G proteins, and cholera and pertussis toxins, like 1, 25(OH)2D3 increased 45Ca2+ uptake by myocytes. AlF4(-) did not further stimulate the effects of 1,25(OH)2D3 thereby showing that a G protein is involved in the hormone action. Moreover, 1,25(OH)2D3 potentiated pertussis toxin but was unable to modify choleric toxin-dependent myocyte Ca2+ influx. Altogether, these results provide evidence indicating that the non-genomic action of 1,25(OH)2D3 on cardiac muscle calcium influx involves modulation of the beta-adrenergic-sensitive adenylyl cyclase/cAMP/PKA pathway coupled to a Gs protein.

Adrenergic beta-Agonists↗
PubMed1999

Rapid actions of calcitriol and its side chain analogues CB1093 and GS1500 on intracellular calcium levels in skeletal muscle cells: a comparative study.

1. The ability of synthetic analogues of the secosteroid hormone 1alpha,25-dihydroxy-vitamin-D3 [calcitriol, CT; 1,25(OH)2D3] to exert non-genomic (rapid) effects on target cells has been scarcely studied. To evaluate the pharmacological potential of the CT side-chain analogues CB1093 and GS1500, we compared their fast effects on intracellular calcium concentration ([Ca2+]i) in chick skeletal muscle cells with those elicited by the natural hormone. 2. Both analogues, similarly to CT, specifically induced rapid (30-60 s) and sustained rises in [Ca2+]i levels. CB1093 and GS1500 were more potent than the natural hormone at concentrations as low as 10(-13) M (4.5 fold stimulation) and 10(-12) M (2.5 fold), respectively, whereas higher concentrations (10(-9)- 10(-8) M) of CT were more effective than the analogues in elevating [Ca2+]i. Cyclic AMP was markedly increased by both analogues pointing for a role of this messenger in the fast actions of the synthetic compounds. 3. In Ca2+ free medium CT and analogues elicited a transient elevation in [Ca2+]i. The PLC inhibitors U73122 (2 microM) and neomycin (0.5 mM), as well as depletion of intracellular stores with thapsigargin (1 microM), completely prevented CB1093/GS1500-dependent changes in [Ca2+]i suggesting that, similarly to CT, these analogues mobilized Ca2+ from an IP3/thapsigargin-sensitive store. 4. The voltage-dependent calcium channel (VDCC) blocker nifedipine (2 microM) reduced by 50-60% the influx phase of the [Ca2+]i response to CB1093 and GS1500, indicating that VDCC contributed partially to Ca2+ entry. The Ca2+ readdition protocol suggested that analogue-dependent activation of a SOC entry pathway accounted, to the same extent as for CT, for the remaining non-VDCC mediated Ca2+ influx.

Animals↗
PubMed1999

Morbidity associated with long-term use of totally implantable ports in patients with AIDS.

To determine the morbidity associated with long-term use of a totally implantable central venous access device (Port-A-Cath [PAC]) in patients with AIDS, we studied 68 consecutive patients with AIDS requiring 79 such devices for long-term use, inserted over a period of 5 years. The total number of PAC-days was 20,159. At least one PAC-related complication occurred with 40 of 79 PACs (50.6% [95% confidence interval (CI): 39.6%-61.6%]), and 16 devices (20.2% [95% CI, 11.4%-29.0%]) had to be removed because of complications. Device-related infection occurred with 33 of 79 PACs (41.7% [95 CI, 30.8%-52.6%]). The predominant infection occurring with PACs was chamber infection, with an incidence of 0.16 per 100 PAC-days. The predominant organisms isolated from patients with chamber infections but also from those with device-related bacteremia were gram-positive cocci (79.4%). The presence of neutropenia (odds ratio [OR] = 9.72; 95% CI, 3.0-31.3; P < .001) and a CD4 cell count lower than 0.025 x 10(9)/L (OR = 6.14; 95% CI, 1.9-19.2; P = .002) were independent predictors of infection. The antibiotic lock technique was associated with decreased device loss when compared with isolated systemic antibiotic therapy (OR = 0.05; 95% CI, 0.0-0.59; P = .008). This technique may be useful to treat PAC infection in patients with AIDS, for whom the risk of PAC-related complications is very high.

AIDS-Related Opportunistic Infections↗
PubMed1999

Acute Q fever in adult patients: report on 63 sporadic cases in an urban area.

We report here 63 sporadic urban cases of acute Q fever diagnosed in 1985-1997. Fifty-eight men and five women were included; the mean age (+/- SD) was 35.6 (+/- 10.2) years. Twenty-six patients had pneumonia, 30 had hepatitis, and 7 had a self-limited febrile illness. The most frequent radiological abnormalities were lobar or segmental alveolar opacities involving right lower lobes. Chronic bronchitis was significantly more frequent among patients with pneumonic Q fever (P = .01). Thirty-two patients' illnesses were diagnosed through seroconversion, 12 by a fourfold increase in serum antibody titer, and 19 by initial high titers. Patients who initially received doxycycline had a significantly shorter duration of fever than those treated with erythromycin or other antibiotics (P = .0001 and P = .0004, respectively). No patient died. Acute Q fever affects mainly urban men, most frequently causing hepatitis, except in those with chronic bronchitis, who more frequently develop pneumonia. Hepatic Q fever presented with more pronounced increases in liver function test values than did pneumonic Q fever. Treatment with doxycycline caused a significant reduction in the duration of fever.

Acute Disease↗
PubMed1998

1alpha,25-dihydroxy-vitamin-D3-induced store-operated Ca2+ influx in skeletal muscle cells. Modulation by phospholipase c, protein kinase c, and tyrosine kinases.

In skeletal muscle cells the steroid hormone 1alpha, 25-dihydroxy-vitamin-D3 (1,25(OH)2D3) nongenomically promotes Ca2+ release from intracellular stores and cation influx through both L-type and store-operated Ca2+ (SOC) channels. In the present work we evaluated the regulation and kinetics of the 1, 25(OH)2D3-stimulated SOC influx in chick muscle cells. Stimulation with 10(-9) M 1,25(OH)2D3 in Ca2+-free medium resulted in a rapid (40-60 s) but transient [Ca2+]i rise, which correlated with sterol-dependent inositol 1,4,5-trisphosphate production. The SOC influx stimulated by the hormone was insensitive to both L-type channel antagonists and polyphosphoinositide-specific phospholipase C (PPI-PLC) inhibitors but was fully inhibitable by La3+ and Ni2+. PPI-PLC blockade prior to 1,25(OH)2D3 stimulation suppressed both the [Ca2+]i transient and the SOC influx. 1,25(OH)2D3-induced SOC entry was markedly increased after 3 min of treatment (30% above basal) and then rapidly reached a steady-state level. The sterol-stimulated SOC influx was prevented by protein kinase C and tyrosine kinase inhibitors but unaffected by blockade of the protein kinase A pathway. None of these inhibitors altered the thapsigargin-induced SOC entry, suggesting the operation of a signaling mechanism different from that for sterol-dependent SOC influx. The present results indicate that 1,25(OH)2D3-induced activation of PPI-PLC is upstream to Ca2+ influx through SOC channels and point for a role of both protein kinase C and tyrosine kinases but not protein kinase A in the regulation of the sterol-dependent SOCE pathway.

Animals↗
PubMed1997

Stimulation of Ca2+ release-activated Ca2+ channels as a potential mechanism involved in non-genomic 1,25(OH)2-vitamin D3-induced Ca2+ entry in skeletal muscle cells.

As in other vitamin D target cells, activation of voltage-dependent Ca2+ channels (VDCC) mediates the fast, non-genomic, 1,25(OH)2D3 stimulation of Ca2+ influx in skeletal muscle cells (SMC). 1,25(OH)2D3 has also been shown to rapidly induce the release of Ins(1,4,5)P3 in SMC. Experiments were performed to investigate whether Ca2+ release-activated Ca2+ channels (CRAC) also participate in the mechanism by which 1,25(OH)2D3 regulates Ca2+ entry into these cells. In cultured chick SMC loaded with Fura-2/AM the hormone (10(-12) - 10(-8) M) induced a rapid (30 sec) followed by a sustained (up to 5 min) increase in intracellular Ca2+ concentration ([Ca2+]i) associated to Ca2+ mobilization from internal stores and influx of extracellular Ca2+, respectively. Thus, the initial, transient, 1,25(OH)2D3-dependent increment in [Ca2+]i could be observed in Ca2+-free medium and was abolished by the PLC inhibitor U73122. Readdition of Ca2+ to cells that had undergone the initial 1,25(OH)2D3-induced [Ca2+]i rise in Ca2+ free medium resulted in a fast increment in [Ca2+]i indicating the existence of a hormone-activated CRAC entry pathway. The sustained phase of the Ca2+ response to 1,25(OH)2D3 was only partially (60%) suppressed by nifedipine, whereas lanthanum (10 microM) completely abolished the hormone effects. Accordingly, depletion of intracellular Ca2+ stores by thapsigargin reproduced 1,25(OH)2D3-induced Ca2+ influx, inhibiting any further response to the sterol. 1,25(OH)2D3 increased the rate of quenching of Fura-2 fluorescence by Mn2+, indicating activation of Mn2+ permeable channels. Altogether, these results provide the first evidence involving CRAC channels in the rapid modulation of Ca2+ entry in animal cells by 1,25(OH)2D3.

Animals↗
PubMed1997

1 alpha,25-(OH)2-vitamin D3 stimulates the adenylyl cyclase pathway in muscle cells by a GTP-dependent mechanism which presumably involves phosphorylation of G alpha i.

To further understand the mechanism underlying 1,25(OH)2D3 activation of the cAMP pathway, the effect of the hormone on adenylyl cyclase (AC), GTPase and protein kinase A (PKA) activities as well as on the phosphorylation of G alpha i was studied in membranes from chick skeletal muscle cells. The sterol stimulated AC activity in a dose (0.1-10 nM) and time (1-5 min.) dependent fashion, provided GTP (10 microM) was present in the assay. High affinity GTPase activity was unaffected by the hormone. In the absence of GTP or in the presence of Mn2+ (20 mM), 1,25(OH)2D3 effects on AC were abolished. PKA activity was increased (+120%) in cells pretreated (1 nM, 5 min.) with the sterol. Moreover, immunoprecipitation of G alpha i from [32P]-labeled myoblast membranes showed that 5 min. exposure to 1 nM 1,25(OH)2D3 increased (1.5-2 fold) the phosphorylation of its alpha subunit. The present data suggest that in muscle cells, 1,25(OH)2D3 activates AC by a non direct, GTP-dependent action which could imply amelioration of Gi function by sterol-induced alpha i phosphorylation.

Adenylyl Cyclases↗
PubMed1997

Comparison of the response to the recombinant vaccine against hepatitis B virus in dialyzed and nondialyzed children with CRF using different doses and routes of administration.

In patients with chronic renal failure (CRF), parenteral transmission of the hepatitis B virus (HBV) is common. The response to the recombinant vaccine is 50%-80% of seroprotection. Therefore, to improve seroprotection, different strategies such as dose augmentation, vaccination at the predialysis stage, subcutaneous application, and using interleukin were tried, with unsatisfactory results. In children, there are no studies demonstrating the efficacy of the vaccine. The aim of this study was to evaluate the efficacy of the recombinant vaccine in children with CRF, in late as well as early phases, through the quantification of antibodies against the surface antigen in response to different doses of the vaccine against HBV. There were 103 patients who were assigned to three groups: (1) 25 patients with CRF in the early phase (undergoing pharmacological treatment only); (2) 67 patients with CRF in the late phase (treatment with peritoneal dialysis or hemodialysis); (3) 11 patients with CRF in the early phase (undergoing low-dose pharmacological treatment only). The antibodies against the serum antigen (HBsAg) were measured by the aEIA method. Urea, creatinine, and creatinine clearance were measured at 0, 2, and 12 months. In our seroprotection results we observed that group 1 and 3 developed earlier seroconversion (50% first month). In patients undergoing dialysis the seroconversion happened in 91% at month 13, but with lower concentration than group 1 and/or group 3 (p < 0.05). In conclusion, there is a better response in predialysis patients. The levels of antibodies are similar in groups 1 and 3 (with small doses), which are similar to the complete doses for an efficient immunity in children with chronic renal failure.

Adolescent↗
PubMed1996

Involvement of protein kinase C in the modulation of 1alpha,25-dihydroxy-vitamin D3-induced 45Ca2+ uptake in rat and chick cultured myoblasts.

The calciotropic hormone 1alpha,25-dihydroxy-vitamin D3 (1,25(OH)2D3) has been shown to stimulate both rat and chick myoblast 45Ca2+ uptake via modulation of dihydropyridine-sensitive L-type calcium channels through phosphorylation by the cAMP/protein kinase A pathway. We further investigated the involvement of protein kinases in 1,25(OH)2D3-signal transduction on cultured myoblasts. The protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) was found to rapidly stimulate myoblast 45Ca2+ uptake, mimicking 1,25(OH)2D3. The effects of PMA were time- (1-5 min) and dose (50-100 nM)-dependent, were mimicked by 1,2-dioctanoylglycerol (DOG) and were specific, since the inactive analogue 4alpha-phorbol was without effect. Analogously to the hormone, PMA-enhanced 45Ca2+ uptake was suppressed by the Ca2+-channel blocker nifedipine (5 microM). 1-(5-isoquinolynsulfonyl)-2-methylpiperazine (H-7), a PKC inhibitor, and down-regulation of PKC by prolonged exposure to PMA (1 microM, 24 h), abolished both PMA and hormone effects on rat and chick cells. As in chick myoblasts, 1,25(OH)2D3 activated PKC in rat myoblasts, with translocation of activity from the cytosol to the cell membrane. Treatment of myoblasts with PMA (100 nM) plus 1,25(OH)2D3 (1 nM) greatly potentiated 45Ca2+ uptake than either agent alone. PMA also increased myoblast cAMP content. These results suggest the involvement of PKC in the mechanism by which 1,25(OH)2D3 rapidly stimulates calcium uptake in both mammalian and avian myoblasts.

Animals↗
PubMed1995

Modulation by 1,25(OH)2-vitamin D3 of the adenylyl cyclase/cyclic AMP pathway in rat and chick myoblasts.

We have previously reported that the calciotropic hormone 1,25(OH)2-vitamin D3 stimulates influx of Ca2+ into cultured rat and embryonic chick myoblasts via voltage sensitive Ca(2+)-channels. In the present study, we show that this effect of 1,25(OH)2D3 requires the mediation of the adenylylcyclase signalling system since the hormone-dependent Ca2+ influx is abolished by specific inhibitors of adenylylcyclase and protein kinase A and mimicked by forskolin and dibutyryl cAMP. 1,25(OH)2D3-stimulated elevations in cellular cAMP paralleled increases in Ca2+ uptake, further suggesting a coupling of adenylylcyclase activation and calcium influx. Fluoride and GTP gamma S mimicked 1,25(OH)2D3-stimulation of calcium influx while GDP beta S suppressed the effect of the hormone. Cholera toxin and Bordetella pertussis toxin both increased 45Ca2+ uptake in rat and chick myoblasts. The hormone further increased cholera toxin actions, but was unable to modify pertussis toxin-induced 45Ca2+ uptake, suggesting a similar target of action for pertussis toxin and 1,25(OH)2D3. Incubation of microsomal membranes with the sterol (10 nM, 2 min) markedly displaces (-32%) [35S]GTP gamma S binding to the membranes. ADP-ribosylation of the pertussis toxin-sensitive 41 kDa substrate was significantly increased (+40%) in 1,25(OH)2D3-pretreated cells. These results suggest that 1,25(OH)2D3-stimulated influx of Ca2+ into rat and embryonic chick cultured myoblasts sequentially requires inhibition of a pertussis toxin-sensitive G protein, accumulation of cAMP and activation of dihydropyridine-sensitive Ca(2+)-channels through PKA-mediated phosphorylation events.

Adenylyl Cyclases↗
PubMed1993

"Stimulation of dihydropyridine-sensitive Ca2+ influx in cultured myoblasts by 1,25(OH)2-vitamin D3".

The rapid, non-genomic effects of 1,25(OH)2-vitamin D3 [1,25(OH)2D3] on calcium influx in cultured neonate rat and embryonic chick myoblasts were investigated. The hormone stimulated 45Ca uptake in a time (1-10 min) and dose (10(-11)-10(-7) M)-dependent manner. The effects of 1,25(OH)2D3 on both rat and chick myoblasts were mimicked by the Ca(2+)-channel agonist BAY K8644 and effectively suppressed by the antagonist nifedipine. 1,25(OH)2D3 induction of Ca2+ uptake was dependent on the presence of extracellular calcium, since it was abolished by prior addition of EGTA and was restored upon the addition of 1 mM Ca2+ Cell membrane depolarization with high K+, similarly to the hormone, elicited a rapid increase in Ca2+ uptake (+75% above control). These results suggest the modulation of Ca2+ influx through competent calcium channels in rat and chick embryo myoblasts by 1,25(OH)2D3.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1989

Mediastinal, left, and right atrial pressure variations with different modes of mechanical and spontaneous ventilation.

Variations in mediastinal, left, and right atrial pressures (MedP, LAP, RAP, respectively) were measured by means of catheters and tubes positioned in ten patients with nonvalvular cardiac surgery. For each pressure, a maximum, minimum, and mean value was determined in relation to its oscillations during the respiratory cycle. Thus, we compared the variations in MedP, LAP, and RAP in controlled mechanical ventilation (CMV), CMV with 5 cm H2O PEEP, synchronous intermittent mandatory ventilation (SIMV), SIMV with 5 cm H2O PEEP, continuous positive airway pressure (CPAP), and spontaneous respiration (SR). We built an experimental model to compare the measurements obtained by air-filled tubes inserted at surgery with those obtained by esophageal balloons filled with water. The maximum MedP did not vary significantly in these patients except when SIMV and SR were compared; however, the minimum MedP diminished significantly (p less than .001) in SIMV, SIMV-PEEP, CPAP, and SR, with negative inspiratory values reaching significant proportions. The mean values of MedP, LAP, and RAP showed a similar tendency although to a lesser degree. The experimental model revealed a strong linear relation between the values obtained with air-filled tubes and those obtained with water-filled esophageal balloons (r = .99, p less than .001). These results suggest that the mean values of MedP, LAP, and RAP do not reflect the dynamic variations in ventricular filling pressure accurately, nor the important negative inspiratory peaks that appear in different types of ventilation using spontaneous cycles with and without PEEP. These inspiratory peaks can overload the left ventricle by hydrostatic gradients, and lead to pulmonary edema in susceptible patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1988

Schistosoma mansoni cationic egg antigens (CEF6): immunoserology with oxamniquine-treated patients and involvement of CEF6 in the circumoval precipitin reaction.

The serologic activity of a cationic fraction (denoted CEF6) of Schistosoma mansoni soluble egg antigen was compared in an ELISA with that of the unpurified soluble egg antigen for the ability to detect human infections and for the prediction of chemotherapeutic success in patients followed up to 5 years post-treatment with oxamniquine. The cationic fraction correctly identified 100% of 20 patients as infected with S. mansoni; moreover, 50% (10 of 20) seroconverted to negative by 2 years post-treatment and 100% of 15 patients tested were negative 5 years post-treatment. In general, the cationic fraction was superior to the unpurified soluble egg antigen for the detection of infection and for the prediction of chemotherapeutic success. The cationic fraction also exhibited greater immunologic specificity over the unpurified soluble egg antigen in that the latter exhibited higher titers than the former to antibodies against heterologous parasite antigens (Fasciola hepatica, Clonorchis sinensis, Paragonimus westermani adult worms; Trichinella spiralis larvae). Moreover, rabbit antisera raised against egg antigens isolated from the precipitation formed when fresh S. mansoni eggs are incubated with S. mansoni infection of immunization sera (known as circumoval precipitin reactions or COP) reacted strongly with the cationic fraction in ELISA. In addition, antiserum to the cationic fraction as well as antisera against either of the two antigenic components of this fraction, known as antigens alpha 1 and omega 1, all give positive COP reactions when incubated with fresh S. mansoni eggs.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗