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Biomedical subjects

M Rubio

Publications and source records attributed to M Rubio.

At least 91 records · Page 5Linked to original sources

Lipid changes in hepatic microsomes and its relationship to P-nitrophenol glucuronidation in an experimental model of portal hypertension.

The liver is responsible for the most important metabolic pathway of non polar compounds. The aim of the present work was to study the p-nitrophenol glucuronidation and its relationship with lipidic composition of microsomal membrane in a model of hepatic portal hypertension and hepatocellular damage induced by monocrotaline. A global increment in liver microsomal phospholipids as well as changes in the phospholipid pattern (phosphatidylethanolamine and sphingomyelin increased up to 156 +/- 13 and 195 +/- 14% respectively) were detected in monocrotaline intoxicated rats when it were compared to control rats. The microsomal cholesterol content showed a decrease in monocrotaline intoxicated rats. (4.1 +/- 0.7 against 6.6 +/- 1.5 micrograms/mg of microsomal protein, in control rats). When p-nitrophenol activity was measured, Km from monocrotaline intoxicated rats was 0.137 mM, and Vmax was 2.9 nmol of p-nitrophenol/mg microsomal protein since in control group Km was 0.322 mM, and Vmax was 4.5 nmol of p-nitrophenol/mg microsomal protein. It is concluded that monocrotaline intoxicated rats showed a different behavior in the kinetics of p-nitrophenol UDP-glucuronyltransferase, as well as a different microsomal lipidic profile, when compared to control group.

Animals↗

Serum tryptase levels in adverse drug reactions.

We evaluated the usefulness of individual tryptase levels and variations after adverse drug reactions in 64 patients. Our aim was to find a tool for the diagnosis of drug allergy. Thirty-seven subjects were confirmed to have drug allergy, 12 had nonsteroidal anti-inflammatory drug (NSAID) reactions, five had negative controlled drug challenges (NAAR), and 10 had symptoms after placebo intake (PLA). Serum tryptase levels greatly increased after anaphylactic shocks (2242%) and anaphylaxis (710.5%). Patients with allergic urticaria and those with idiosyncratic responses to acetylsalicylic acid (ASA) exhibited a small increase in serum tryptase (49.5% and 38.2%, respectively). In the other two groups (NAAR and PLA), no variation in this serum protease was observed. The time of appearance of the serum tryptase peak differed considerably among patients with similar clinical reactions (from 30 min to 6 h) and was independent of the latent period, severity of symptoms, or the amount of tryptase released. We conclude that serum tryptase determinations are helpful in the diagnosis of anaphylactic shock and anaphylaxis, but serial measurements may be needed to confirm mast-cell participation in milder reactions.

Anaphylaxis↗

bldA-dependent expression of the Streptomyces exfoliatus M11 lipase gene (lipA) is mediated by the product of a contiguous gene, lipR, encoding a putative transcriptional activator.

Extracellular lipase synthesis by Streptomyces lividans 66 carrying the cloned lipase gene (lipA) from Streptomyces exfoliatus M11 was found to be growth phase dependent, since lipase was secreted into the medium mainly during the stationary phase; S1 nuclease protection experiments revealed abundant lipA transcripts in RNA preparations obtained during the stationary phase but not in those obtained during exponential growth. Transcription from the lipA promoter was dependent on the presence of lipR, a contiguous downstream gene with a very high guanine-plus-cytosine content (80.2%). The deduced lipR product consists of a protein of 934 amino acids that shows similarity to known transcriptional activators and has a strong helix-turn-helix motif at its C terminus; this motif is part of a domain homologous to DNA-binding domains of bacterial regulators of the UhpA/LuxR superfamily. The lipR sequence revealed the presence of a leucine residue, encoded by the rare TTA codon, which caused bldA dependence of lipA transcription in Streptomyces coelicolor A3(2); replacement of the TTA codon by the alternate CTC leucine codon alleviated bidA dependence but not the apparent growth phase-dependent regulation of lipA transcription. When lipR expression was induced in a controlled fashion during the exponential growth phase, by placing it under the inducible tipA promoter, lipase synthesis was shifted to the exponential growth phase, indicating that the timing of lipR expression, and not its bldA dependence, is the main cause for stationary-phase transcription of lipA.

Amino Acid Sequence↗

Decreases in the size and proliferation rate of VIP-immunoreactive cells induced in vitro by testosterone are associated with decreases in VIP release.

As in vitro study was carried out to determine whether testosterone directly inhibits the secretion of pituitary vasoactive intestinal peptide (VIP). A study was also conducted to elucidate the repercussions of treatment with 10(-6) M testosterone of monolayer cultures in the size and proliferation rate of pituitary VIP-immunoreactive cells, using double immunocytochemical labelling for proliferating cell nuclear antigen (PCNA) and VIP. Testosterone decreased the in vitro release of VIP from pituitary cells. It also decreased the percentage of pituitary VIP-immunoreactive and PCNA- and VIP-immunoreactive cells and decreased the size of the cellular and nuclear areas of these cells. Our results suggest that the inhibition of pituitary VIP secretion induced by testosterone could be exerted at pituitary level and that testosterone could act as a modulator of the proliferation rate of rat pituitary VIP-immunoreactive cells.

Animals↗

Corticosterone induces hypoactivity of prolactin-immunoreactive cells.

In order to determine whether corticosterone regulates activity of rat lactotrophs by acting directly at the pituitary level, immunohistochemical studies were carried out in adrenalectomized rats, subjected or not to treatment with corticosterone or colchicine, and in monolayer cultures after incubation with corticosterone. Adrenalectomy increased cellular and nuclear areas (p<0.01) of prolactin-immunoreactive cells without affecting their cytoplasmic area. Similar results were found in adrenalectomized and colchicine-treated animals. Corticosterone reversed the effects of adrenalectomy, although normal values were partially reversed. In cultured pituitary cells, exposure to corticosterone reduced numerical density and cellular, cytoplasmic and nuclear areas with respect to control dishes. Morphological differences in shape, arrangement and nuclear features were observed after treatment with corticosterone. These results demonstrate an inhibitory effect of corticosterone on the activity of rat pituitary prolactin cells and suggest that corticosterone induces hypoactivity by acting on the pituitary prolactin cells of male rats.

Adrenalectomy↗

In vitro effects of estradiol on pituitary GH-immunoreactive cells.

In order to determine whether estradiol modulates the proliferation and activity of somatotrophic cells in vitro, a study of GH-immunoreactive cells was carried out in pituitary monolayer cultures obtained from male adult rats treated with 10(-6)M estradiol for 3 hours. Cellular activity was evaluated in a morphometric study of GH-immunoreactive cells. The proliferation rate was determined by double immunostaining for GH and PCNA (proliferating cellular nuclear antigen). The results were compared to those obtained from control dishes. Estradiol was seen to increase the cellular (p < 0.05), nuclear (p < 0.01) and cytoplasmic areas (p < 0.05). Estradiol decreased the percentage of proliferating GH-immunoreactive cells (p < 0.05) and the nuclear density of somatotrophs (p < 0.05) when all cells present in the dishes were considered. However, when only GH-immunoreactive cells were considered, estradiol increased the proliferation rate of these cells (p < 0.05). Overall, our results suggest that, in vitro, estradiol stimulates the cellular activity and proliferation of GH-immunoreactive cells in the rat.

Animals↗

The pharmacokinetics of ceftazidime in lactating and non-lactating cows.

The pharmacokinetics of ceftazidime (CAZ) were studied in lactating (LTG) and non-lactating (NLTG) cows. Two groups (LTG and NLTG) of 5 healthy dairy cows were given ceftazidime (10 mg/ kg body weight) intravenously (i.v.) and intramuscularly (i.m.). Serum and milk (LTG) and serum samples (NLTG) were collected over a 24-h period post-administration. CAZ concentrations in serum and milk were determined by high-performance liquid chromatography, and an interactive and weighted-non-linear least-squares regression analysis was used to perform the pharmacokinetic analysis. The pharmacokinetic profiles in LTG and NLTG cows which had received CAZ i.v. fitted a three-compartment model and a two-compartment model, respectively. The CAZ concentration-time curves in serum and the area under the curve were greater and more sustained (p < 0.05) in the LTG cows by both routes, while the serum clearance (Cls = 72.5 +/- 18.1 ml/h per kg) was lower (p < 0.05) than that in the NLTG cows (Cls = 185.9 +/- 44.2 ml/h per kg). CAZ given i.v. exhibited a relatively long half-life of elimination (t1/2 beta (LTG) = 1.1 +/- 0.2 h; t1/2 beta (NLTG) = 1.4 +/- 0.3 h). Compared with other cephalosporins, CAZ had good penetration into the mammary gland (47.7 +/- 38.2% for CAZ i.v.; 51.1 +/- 39.0% for CAZ i.m.). Finally, the bioavailability of CAZ (F(LTG) = 98.9 +/- 36.8%; F(NLTG) = 77.1 +/- 25.3%) was suitable for its used by the i.m. route in lactating and non-lactating cows.

Animals↗

NADPH-diaphorase activity and vasopressin in the paraventricular nucleus of the hypothalamus following adrenalectomy.

In order to investigate novel neuroendocrine functions of the nitric oxide synthesizing enzyme a combined histochemical and immunocytochemical study focused on the paraventricular nucleus of the hypothalamus was conducted to check a possible influence of bilateral adrenalectomy on three different neuronal populations, NADPH diaphorase (ND)-positive, vasopressin (VP)-immunoreactive and neurons expressing both markers. In the adrenalectomized animals, a slight increase (P > 0.05) of the number of ND magnocellular neurons was detected, whereas no changes were observed in the ND-parvicellular population and in the neurons showing coexistence (magno- and parvicellular) (P > 0.05). By contrast, following bilateral adrenalectomy, a significant increase (P < 0.05) in the VP-parvicellular population (anterior, medial and periventricular subdivisions) was detected, which was reversed when the animals received daily doses of corticosterone. These results suggest that nitric oxide is not closely related to the hypothalamic regulation of the adenocorticotropin secretion exerted by the paraventricular nucleus.

Adrenal Glands↗

Dopamine inhibits in vitro release of VIP and proliferation of VIP-immunoreactive pituitary cells.

A double immunohistochemical study for VIP and proliferating cell nuclear antigen (PCNA) was carried out on monolayer cultures from adult male rats pituitary glands treated with dopamine (ranging from 10(-9) to 10(-5) M), in order to establish whether or not dopamine is involved in the regulation of the proliferation rate of pituitary VIP-immunoreactive cells. For all doses of dopamine assayed, the release of VIP to the culture medium, the numerical density of VIP-immunoreactive cells and the percentages of VIP- and PCNA-immunoreactive cells decreased significantly after dopamine treatment. These results suggest that dopamine could be a physiological inhibitor involved in the modulation of pituitary VIP proliferation rate.

Animals↗

Effects of S-adenosylmethionine on lipid peroxidation and liver fibrogenesis in carbon tetrachloride-induced cirrhosis.

BACKGROUND/AIM: The aim of this study was to investigate the effects of S-adenosylmethionine on liver peroxidation and liver fibrogenesis in carbon tetrachloride-induced cirrhosis. METHODS: Cirrhosis was induced in three groups of six rats by repeated injections of carbon tetrachloride over 9 weeks. One group of animals was treated only with carbon tetrachloride, and the other two received carbon tetrachloride plus S-adenosylmethionine (10 mg/kg intramuscularly daily) from week 3 to week 9, and from week 6 to week 9 of the study, respectively. Two additional groups of six rats, a control group and a group treated only with S-adenosylmethionine, were also studied. Glutathione concentration, thiobarbituric acid-reactive substances, collagen content, prolyl hydroxylase activity, and procollagen type I mRNA expression were determined in liver samples. RESULTS: All carbon tetrachloride-treated rats had cirrhosis at the end of the study. Cirrhosis was also present in five of the six carbon tetrachloride-treated rats receiving S-adenosylmethionine for 3 weeks, but in only one of the six rats that received S-adenosylmethionine for 6 weeks. Hepatic glutathione was significantly diminished in carbon tetrachloride-treated rats (2.7 +/- 0.3 mumol/g tissue) and returned to normal in rats receiving S-adenosylmethionine for 3 or 6 weeks (3.7 +/- 0.13 and 3.9 +/- 0.11 mumol/g tissue, respectively). The hepatic thiobarbituric acid-reactive substances were significantly lower in rats treated with carbon tetrachloride and S-adenosylmethionine for 6 weeks (98 +/- 5 nmol/g) than in rats treated with carbon tetrachloride (134 +/- 12 nmol/g) and in those treated with carbon tetrachloride and S-adenosylmethionine for 3 weeks (127 +/- 13 nmol/g). There were no differences in either hepatic collagen and prolyl hydroxylase activity between rats that received only carbon tetrachloride and those treated with S-adenosylmethionine for 3 weeks. In contrast, carbon tetrachloride-treated rats receiving S-adenosylmethionine for 6 weeks had significantly lower collagen and prolyl hydroxylase activity than the other two groups. A much greater increase in procollagen type I mRNA was found in carbon tetrachloride-treated rats than in rats treated with carbon tetrachloride and S-adenosylmethionine for 6 weeks. Furthermore, there was a significant correlation between the hepatic thiobarbituric acid-reactive substances and prolyl hydroxylase activity and hepatic collagen. CONCLUSIONS: We conclude that the early administration of S-adenosylmethionine in a model of carbon tetrachloride-induced liver injury restores glutathione levels and reduces lipid peroxidation, resulting in less advanced liver fibrosis.

Animals↗

Pharmacokinetics of Dextran-70 in patients with cirrhosis and ascites undergoing therapeutic paracentesis.

BACKGROUND/AIM: Dextran-70 is frequently used as a plasma expander in patients with cirrhosis treated with large-volume paracentesis to prevent post-paracentesis hypovolemia, which is thought to develop after 24 h following the procedure. However, there are no studies on Dextran-70 pharmacokinetics in cirrhosis. METHODS: Nine patients with alcoholic cirrhosis and tense ascites treated with a 5-1 paracentesis were given 500 ml of Dextran-70. Blood samples to measure the plasma concentration of dextran were obtained 15 and 30 min, 1, 2, 3, 6, 12 and 24 h and 2 and 6 days after the end of the infusion. Nine healthy volunteers were studied in identical fashion after infusion of 100 ml of Dextran-70. The plasma concentration of dextran was determined by the anthrone method. A bicompartmental model was used to analyze the pharmacokinetic parameters. RESULTS: There were no significant differences between patients with cirrhosis and controls in the volume of distribution (7.7 +/- 0.6 vs. 7.3 +/- 1.21), half-life of the first and second component of plasma disappearance (2.96 +/- 0.69 and 80.3 5.9 h in patients with cirrhosis vs 2.82 +/- 0.69 and 67.1 +/- 10.7 h in controls). CONCLUSIONS: The pharmacokinetics of Dextran-70 in patients with cirrhosis and ascites after large-volume paracentesis is similar to that in controls. This may explain why Dextran-70 is less effective than albumin in preventing paracentesis-induced hypovolemia, which starts after most Dextran fraction has disappeared from plasma.

Ascites↗

Nosocomial outbreak of Burkholderia pickettii infection due to a manufactured intravenous product used in three hospitals.

Forty-six cases of nosocomial infection caused by Burkholderia pickettii were reported between June and November 1993 in three metropolitan hospitals in Madrid. A case-control study of the outbreak was conducted to identify its cause. Seventy-four percent of the patients were males; the mean age +/- SD of the patients was 54 +/- 20 years. Sixty-five percent of the patients presented with some gastrointestinal disorder, and 80% had a peripheral catheter; 98% were treated with intravenous fluids, and 96% were treated with intravenous ranitidine. On the basis of results of a descriptive study and knowledge of the epidemiologic features of B. pickettii, a provisional causal hypothesis was formulated: intravenous ranitidine was the source of the outbreak. As a control measure, it was advised to stop treatment with this drug. On the basis of results of logistic regression and the microbiological isolation of B. pickettii in an ampule of the drug, we concluded that intravenous ranitidine was the cause of the outbreak.

Adult↗

A study of Light's criteria and possible modifications for distinguishing exudative from transudative pleural effusions.

STUDY OBJECTIVE: To determine the usefulness of modifying Light's criteria for the separation of pleural transudates from exudates. DESIGN: Retrospective review of patients who underwent a diagnostic thoracentesis during a 2-year period. SETTING: Community teaching hospital in Lleida, Spain. PATIENTS AND METHODS: Clinical records and pleural fluid characteristics of 230 consecutive patients with pleural effusion underwent a detailed review. Thirty-five of these patients were excluded from the analysis. As suggested recently by Romero et al, different cutoff levels for the criteria of Light et al were applied and their accuracies were calculated. RESULTS: Thirty-nine (20%) pleural effusions were transudates and 156 (80%) were exudates. The accuracy of the criteria of Light et al for identifying exudates was 94.7% (confidence interval, 91.6 to 97.9) in comparison to our own modified criteria (93.1%; confidence interval, 89.5 to 96.7) and the criteria suggested by Romero et al (92.6%; confidence interval, 88.9 to 96.3). These differences were not statistically significant. CONCLUSION: Changing the classic Light's criteria with different cutoff points offers no advantages for discriminating between transudative and exudative pleural effusions.

Adolescent↗

Morphometric evidence of the regulation of the activity of LH-immunoreactive cells by corticosterone. An in vivo and in vitro study.

The present study was performed to investigate the morphometric effects of the administration of corticosterone on LH-immunoreactive cells of pituitary monolayer cultures. In addition, the effect of bilateral adrenalectomy on the LH-adenohypophyseal cells was explored. In vitro administration of corticosterone induced a marked decrease in the morphometric parameters considered (cellular, cytoplasmic and nuclear areas). In vivo, bilateral adrenalectomy produced a significant increase in these parameters when compared to those obtained in normal and sham-operated animals. However, when the bilaterally adrenalectomized rats received a daily dose of corticosterone no significant changes in the morphometric parameters were found. Morphometrically, these combined in vivo-in vitro findings confirm a corticosterone-induced hypoactivity of LH-cells together with a direct effect of this glucocorticoid on LH-adenohypophyseal cells.

Adrenalectomy↗