Search PubMed⌕ Search

Biomedical subjects

J M Silva

Publications and source records attributed to J M Silva.

At least 91 records · Page 5Linked to original sources
PubMed1993

Molecular mechanisms of nitrofurantoin-induced hepatocyte toxicity in aerobic versus hypoxic conditions.

We have previously reported that incubation of isolated rat hepatocytes with the antimicrobial drug nitrofurantoin causes toxic oxidative stress as a result of oxygen activation by futile one-electron redox cycling. In continuation of this work nitrofurantoin was found to cause rapid glutathione-protein mixed disulfide formation before GSSG had completely effluxed from the hepatocytes. Furthermore, the disulfide reductant dithiothreitol added up to 60 min later restored protein thiols and prevented cytotoxicity. In contrast, under hypoxic conditions nitrofurantoin-induced hepatocyte toxicity was associated with [14C]nitrofurantoin irreversible binding to protein, protein thiol depletion without glutathione-protein mixed disulfide formation, and GSH depletion. Furthermore, dithiothreitol did not have any antidotal properties and did not reverse covalent binding though some binding was prevented. These results suggest that under aerobic conditions nitrofurantoin-induced hepatocyte toxicity is due to the formation of glutathione-protein mixed disulfides, while under hypoxic conditions toxicity is associated with protein alkylation.

Aerobiosis↗
PubMed1993

Insertional inactivation of the WT1 gene in tumour cells from a patient with WAGR syndrome.

The WT1 gene was analysed using DNA from a Wilms' tumour derived from a patient with the WAGR syndrome using single strand conformation polymorphism analysis and polymerase chain reaction sequencing. A 14-bp insertion was found in the intron part of the splice donor site of exon 7 and was a tandem duplication of an upstream exon sequence. This mutation would be expected to disrupt the correct processing of the WT1 mRNA and is predicted to result in a non-functional protein. This observation further supports the role of WT1 in Wilms' tumorigenesis in patients with constitutional 11p13 deletions.

Abnormalities, Multiple↗
PubMed1993

Molecular mechanisms of SR 4233-induced hepatocyte toxicity under aerobic versus hypoxic conditions.

SR 4233 (3-amino-1,2,4-benzotriazine-1,4-dioxide) is the lead compound of the benzotriazene-di-N oxides which are selectively toxic to tumour cells under hypoxic conditions. However much higher concentrations given to rats caused bone marrow toxicity and necrosis of the low oxygen Zone 3 part of the liver. In the following effects of SR 4233 on hepatocytes under hypoxic vs aerobic conditions have been compared. (1) SR 4233 did not affect hepatocyte viability (as determined by plasma membrane disruption) or glutathione levels under aerobic conditions. SR 4233 however induced cyanide-resistant respiration, an indicator of redox cycling mediated oxidative stress and became cytotoxic if hepatocyte catalase or glutathione reductase was inactivated. Glutathione oxidation occurred well before cytotoxicity ensued. Addition of ascorbate markedly enhanced SR 4233 cytotoxicity to these compromised hepatocytes. (2) In contrast, SR 4233 was highly toxic to hypoxic hepatocytes. Addition of ascorbate to enhance SR 4233 reduction also caused a marked increase in hepatocyte toxicity and an SR 4233 radical was detected with ESR spectroscopy. SR 4233 cellular reduction and toxicity was prevented with fructose or inhibitors of NADPH:cytochrome P-450 reductase. Inactivation of catalase or glutathione reductase had no effect on SR 4233 toxicity and hepatocyte GSH was not oxidised indicating oxidative stress did not occur during hypoxic SR 4233 hepatocyte toxicity. (3) The lack of SR 4233 cytotoxicity under aerobic conditions could probably be attributed to the detoxification of the SR 4233 radical by mitochondrial oxidation as SR 4233, but not its metabolite SR 4317 markedly increased state III and IV mitochondrial respiration in the presence of NADH. The increased respiration was inhibited by the respiratory inhibitors KCN and antimycin A but not by rotenone. Furthermore SR 4233 cytotoxicity under aerobic conditions was markedly increased by partially inhibiting hepatocytes respiration with cyanide but not rotenone.

Animals↗
PubMed1992

Modulation of trenimon-induced cytotoxicity by DT-diaphorase in isolated rat hepatocytes under aerobic versus hypoxic conditions.

Trenimon belongs to a class of aziridinylbenzoquinone anticancer drugs that cross the blood-brain barrier. In this study we have investigated the molecular mechanisms for trenimon-induced toxicity in aerobic versus hypoxic conditions with the use of freshly isolated rat hepatocytes. The following evidence suggests the mechanisms for trenimon detoxification involves reduction by DT-diaphorase, while the cytotoxic mechanism involves macromolecular alkylation under hypoxic conditions as well as oxidative stress under aerobic conditions. (a) Hepatocyte cytotoxicity induced by trenimon (250 microM) under aerobic conditions ensued following an initial induction of cyanide-resistant respiration and partial oxidation of glutathione to oxidized glutathione. Trenimon reduction to the hydroquinone by the hepatocytes was rapid. Inhibition of hepatocyte DT-diaphorase by dicumarol increased trenimon-induced cytotoxicity by approximately 10-fold, and markedly inhibited hydroquinone formation. Furthermore, both cyanide-resistant respiration and oxidized glutathione formation were markedly increased, resulting in depletion of oxygen in the media. Trenimon reduction to the hydroquinone then occurred. This suggests that DT-diaphorase in normal hepatocytes prevents the formation of the semiquinone that causes cytotoxic protein alkylation and oxidative stress. (b) Hepatocyte cytotoxicity induced by trenimon (350 microM) under hypoxic conditions ensued following glutathione depletion without oxidized glutathione formation. Inactivation of hepatocyte DT-diaphorase by dicumarol under hypoxic conditions increased trenimon-induced cytotoxicity by approximately 3.5-fold and increased semiquinone radical levels 2-fold without affecting its reduction rate. This suggests that the cytotoxic mechanism involves protein alkylation by semiquinone radicals formed by reductases catalyzing a one-electron reduction of trenimon.

Aerobiosis↗
PubMed1992

Magnetic field exposure assessment for adult residents of Maine who live near and far away from overhead transmission lines.

Sixty-Hz magnetic field exposures were measured for 45 adult residents of Maine. Thirty of the subjects resided near rights-of-way (ROWs) with either 345- and 115-kV transmission lines, or ROWs with only 115-kV transmission lines; fifteen resided far from any transmission lines. Personal exposure data for a single 24-hour period was acquired with the EMDEX. The EMDEX's event-marker button was used to partition exposures into Home and Away components. Also, three area measurements were taken for each subject during the personal exposure measurement period: 1) 24-hr fixed-site bedroom measurement with a second EMDEX; 2) Spot measurements in at least three rooms of every residence; and 3) Spot measurements outside each residence. Residence near transmission lines highly loaded during the measurement period was associated with increased Home and Total exposure relative to a far-away population. Average exposure level while away from home was uniform (at about 2 mG) throughout the study population. On a quantitative level, Home exposure was correlated equivalently with Spot-In (r = .70) and the 24-hr fixed site measurement (r = .68). Correlations of area measurements with Total exposure were weaker because of the dilution effect of Away exposure (r = .64 for Spot-In; r = .61 for 24-h Bedroom). Away and Home exposures were not correlated (r = .14), which reinforced our confidence that the participants used the EMDEX correctly. The data suggest the need for caution before inferences are drawn about total personal exposure from area measurements. The study demonstrates the feasibility of obtaining valid measures of magnetic-field exposure with the personal exposure monitors that have been developed.

Adult↗
PubMed1992

Small integrating meter for assessing long-term exposure to magnetic fields.

A small, lightweight meter has been developed for magnetic-field measurements, particularly those needed for exposure-assessment purposes. This meter, known as the AMEX-3D, continuously measures all three axes of magnetic-flux density and electronically combines the data into a single estimate of cumulative exposure to the root-mean-square (rms) resultant flux density. The AMEX-3D weighs about 120 g, measures 2.7 cm x 5.1 cm x 10.2 cm, and is battery powered. Two panel-mounted jacks are provided for measuring battery voltage and for reading cumulative exposure data from the unit. The instrument has, within 3 dB, a flat response to magnetic flux densities at all frequencies in its 30-1,000 Hz bandwidth. A detailed analysis of error sources in the AMEX-3D leads to an estimate of +/- 20% as the accuracy of the instrument over its dynamic range, which extends from 0.02 to 15 microT. The AMEX-3D was tested in the field by asking electric-utility distribution linemen to wear AMEX-3D and EMDEX meters simultaneously while working. Agreement between the two measures of exposure was excellent.

Electromagnetic Fields↗
PubMed1992

Molecular mechanisms of trenimon-induced cytotoxicity in resistant L5178Y/HBM10 cells.

L5178Y/HBM10 lymphoblasts, resistant to a model quinone antitumor agent, hydrolyzed benzoquinone mustard, were approximately 2-fold more sensitive to trenimon (2,3,5-tris-ethyleneimino-1,4-benzoquinone) compared to parental cells (L5178Y). The L5178Y/HBM10 cells are reported to have a 24-fold increased level of DT-diaphorase activity over the parental cells. Inhibition of DT-diaphorase by dicoumarol markedly inhibited the cytotoxic activity of trenimon to the resistant L5178Y/HBM10 cells. Spectrophotometric analysis of the reduction of the quinone, trenimon, to its hydroquinone form was shown to occur approximately 25 times more rapidly in the L5178Y/HBM10 cells relative to the parental cells and was inhibited by discoumarol. Trenimon also induced continuous cyanide-resistant respiration in the L5178Y cells, but not in the resistant L5178Y/HBM10 cells. This suggested a one-electron reduction of trenimon to a semiquinone free radical which could then redox cycle with oxygen in the L5178Y cells. However, in the presence of dicoumarol the resistant L5178Y/HBM10 cells induced similar oxygen activation to the parental cells. Dicoumarol had no effect on trenimon-induced cyanide resistant respiration in the parental cells. These findings suggest that the two-electron reduction of trenimon to its hydroquinone derivative plays a major role in the cytotoxic activity of trenimon.

Cell Survival↗
PubMed1992

[Relations between the levels of lipoprotein (a) and results of coronarography in patients with ischemic heart disease. Preliminary results].

The authors make a short revision about Lipoprotein(a) [Lp(a)], which has been referred as an independent risk factor for coronary disease. To confirm this results in a portuguese population, we studied serum Apo(a) [method IRMA.Kit Pharmacia.1 U/L Apo(a) = 1 mg/L Lp(a)], Apo A, Apo B, total cholesterol, HDL-cholesterol, triglycerides, calcium, uric acid, glucose, fibrinogen and ESR in 37 patients who were submitted to postmyocardial infarction coronarography (Judkins technic--Seldinger. Classic incidences. Significative lesions: greater than or equal to 75% in coronary vessels or greater than or equal to 50% in common main nk). The purpose was to search for changes on the plasmatic levels according to the results of the coronarography. The only change observed was in the Lp(a) levels, with important variations (p less than 0.05) between the defined groups: patients without significant coronary disease or disease on just one vessel [media (mean) = 30.1 mg/dl. Standard error (SE) = 7.9 mg/dl.n = 11] and another group of patients with coronary disease of two or three vessels [mean = 50.5 mg/dl.SE = 7.0 mg/dl.n = 26]; among the patients with two vessels disease were found significant changes (p less than 0.02) on Lp(a) levels according to the descending anterior coronary being [mean = 79.5 mg/dl.SE = 4.3 mg/dl.n = 8] or not [mean = 38.5 mg/dl.SE = 14.3 mg/dl.n = 7] implicated. These observations make the suggestion of Lp(a) as a risk factor for coronary disease, and a predictor for the gravity of the coronary disease in myocardial infarction.

Adult↗
PubMed1991

Prevention of nitrofurantoin-induced cytotoxicity in isolated hepatocytes by fructose.

Nitrofurantoin is a widely utilized urinary antimicrobial drug which has been associated with pulmonary fibrosis, neuropathy, and hepatitis as well as hemolytic anemia in glucose-6-phosphate dehydrogenase-deficient individuals. Incubation of freshly isolated rat hepatocytes with nitrofurantoin caused oxygen activation as a result of futile redox cycling. Glutathione disulfide (GSSG) was formed and rapidly exported from the cell resulting in complete glutathione (GSH) depletion followed by cell death. However, fructose prevented the export of GSSG from the cell and GSH levels recovered rapidly without cytotoxicity occurring. Fructose did not affect nitrofurantoin metabolism but rapidly depleted cellular ATP levels by approximately 80% which remained depressed during the incubation period. Fructose, however, did not protect hepatocytes from nitrofurantoin-induced cytotoxicity if GSH was depleted beforehand. Protection by fructose only occurred at concentrations which caused ATP depletion. These results suggest that fructose prevents nitrofurantoin-induced toxicity by depleting ATP and thereby preventing the ATP-dependent GSSG efflux. GSSG is retained enabling NADPH and glutathione-reductase to reduce the GSSG back to GSH, thereby protecting the cell from nitrofurantoin-induced oxidative stress.

Adenosine Triphosphate↗
PubMed1989

Diaziquone-induced cytotoxicity in isolated rat hepatocytes.

2,5-Diaziridinyl-3,6-bis(carboethoxyamine)-1,4-benzoquinone (AZQ) is a lipid-soluble antitumor agent. The following evidence using isolated rat hepatocytes as a model for cytotoxicity studies suggests that, under aerobic conditions, AZQ participates in futile oxidation-reduction cycling and oxygen activation. The H2O2 formed can mediate oxidative stress cytotoxicity in compromised cells. (a) Addition of AZQ to hepatocytes causes the stoichiometric oxidation of glutathione (GSH) to oxidized glutathione. No subsequent reduction back to GSH occurred. This was found to be the result of reversible inactivation of glutathione reductase by AZQ. The extent of GSH oxidation increased with AZQ concentration. (b) Cytotoxicity occurred when AZQ concentrations were sufficient to completely deplete GSH. (c) Addition of AZQ to hepatocytes enhanced cyanide-resistant respiration. (d) If the hepatocytes were compromised with azide or cyanamide to inhibit catalase, cytotoxicity was increased 10-fold or 100-fold if ascorbate was also present. (e) AZQ readily induced Ca2+ release by energized mitochondria. Ascorbate markedly enhanced the effectiveness of AZQ, and catalase delayed Ca2+ release. H2O2 formed by aerobic oxidation-reduction cycling of AZQ may therefore cause mitochondrial Ca2+ release.

Animals↗
PubMed1989

Allyl alcohol- and acrolein-induced toxicity in isolated rat hepatocytes.

Incubation of isolated hepatocytes with allyl alcohol results in GSH depletion and subsequent cytotoxicity which is prevented by pyrazole, an inhibitor of alcohol dehydrogenase. Both GSH depletion and cytotoxicity were much more rapid when hepatocytes were incubated with acrolein, the reactive metabolite, and were not affected by pyrazole. However, cytotoxicity of both allyl alcohol and acrolein was enhanced by the aldehyde dehydrogenase inhibitors cyanamide and disulfiram. Malondialdehyde, a lipid peroxidation product, was also formed when hepatocytes were incubated with either agent, and treatment of the hepatocytes with a ferric ion chelator, desferrioxamine, or an antioxidant delayed the cytotoxicity without affecting GSH depletion. Although no GSSG was formed and addition of disulfide reductant dithiothreitol did not restore GSH levels, cytotoxicity was prevented if dithiothreitol was added some time after either agent.

1-Propanol↗
PubMed1989

Diethyldithiocarbamate (DEDC) enhances quinone mediated oxidative stress cytotoxicity in isolated hepatocytes by forming toxic quinone conjugates.

The copper-chelating thiol drug, diethyldithiocarbamate (DEDC) had previously been used to inhibit superoxide dismutase (SOD) and enhance oxidative stress mediated cytotoxicity. Using isolated rat hepatocytes, it was confirmed that DEDC enhances oxidative stress cytotoxicity induced by 1,4-naphthoquinone (1,4-NQ) and 1,4-naphthoquinone-2-sulphonate (1,4-NQ-2S). However, equimolar concentrations of DEDC also enhances cytotoxicity induced by benzoquinone, previously shown to cause cytotoxicity as a result of alkylation and not oxidative stress. Higher DEDC concentrations on the other hand protected against benzoquinone-induced cytotoxicity. Finally, the susceptibility of hepatocytes to quinone mediated oxidative stress cytotoxicity was not enhanced if the DEDC was removed before incubating the hepatocytes with naphthoquinone or benzoquinone. Enhanced oxidative stress cytotoxicity was only observed if the DEDC was present when hepatocytes were treated with quinones. It was concluded that DEDC forms conjugates with quinones which undergo futile redox cycling in the hepatocyte and form H2O2 as well as increase the susceptibility of hepatocytes to H2O2.

Animals↗
PubMed1989

[Percutaneous mitral valvotomy].

We report the first percutaneous mitral valvotomy performed in Portugal in October 19th 1987. The valvotomy was attempted successfully in a 34 year old man, with calcific mitral stenosis, in NYHA class III, with functional mitral valve area less then 1 cm2. The intervention was performed using the transseptal technique with two balloons (20 and 18 mm in diameter), placed in the mitral valve annulus through two long transseptal sheaths (14F). The procedure resulted in a marked decrease in the diastolic transmitral gradient, and an increase in mitral valve area superior to 100%. No immediate or late complications were observed, namely mitral regurgitation or left to right shunt through the atrial septum. One year later the patient is in NYHA class I, with a mitral valve area of 1.7 cm2.

Adult↗
PubMed1989

[Pulmonary valvotomy in adults. The double balloon technic--apropos of 2 cases].

Percutaneous double balloon valvotomy was performed in two adult patients with severe non dysplasic pulmonary valvular stenosis. The balloons were introduced through two 14F sheaths placed in the right and left femural veins, the intervention was well tolerated and succeeded with a marked decrease in right ventricular pressure and in pulmonary transvalvular gradient (113 and 147 mmHg to 10 and 12 mmHg respectively). The patients were discharged within 24 hours, without the need of Intensive Care admission. Indications for the procedure and technical aspects are presented, as well as the potential advantages of two balloons over one balloon and surgical valvotomy.

Catheterization↗
PubMed1988

Nitrofurantoin-mediated oxidative stress cytotoxicity in isolated rat hepatocytes.

Freshly isolated rat hepatocytes were used to study the mechanism(s) of toxicity of the antimicrobial drug nitrofurantoin. This 5-nitrofuran derivative stimulated hepatocyte oxygen uptake in the presence of the mitochondrial respiration inhibitors KCN or antimycin A. This could indicate the formation of O2- and H2O2, following intracellular nitrofurantoin reduction. Addition of nitrofurantoin to suspensions of isolated rat hepatocytes produced a dose- and time-dependent decrease of cell viability. H2O2 probably plays a significant role in the cytotoxic effects of nitrofurantoin as the catalase inhibitors azide or aminotriazole markedly enhanced cytotoxicity. The loss of cell viability was preceded by glutathione (GSH) depletion and a concomitant and nearly stoichiometric formation of oxidised glutathione (GSSG) that did not occur in hepatocytes lacking glutathione peroxidase activity isolated from rats fed a low-selenium diet. This indicates that H2O2 and the seleno-enzyme glutathione peroxidase are responsible for GSH oxidation. Furthermore, addition of nitrofurantoin to isolated rat hepatocytes produced a reversible inactivation of hepatocyte glutathione reductase activity and explains the maintenance of high GSSG levels. The compromised hepatocytes were also highly susceptible to H2O2. The hepatocyte toxicity of nitrofurantoin may, therefore, be attributed to oxidative stress caused by redox-cycling mediated oxygen activation.

Animals↗
PubMed1986

The relationship between selected psychological traits and fear of success in senior elite level wrestlers.

The present study sought to examine the relationship between selected psychological traits as measured by the Institute for Personality and Ability Testing (IPAT) 16 PF inventory and fear of success as measured by the Fear of Success Scale (FOSS). The subjects were 36 Senior elite wrestlers (age 20-25) who were invited to the Colorado Springs Olympic Training Center to prepare for an up-coming inter-national competition. A stepwise multiple regression analysis revealed that the following 16 PF factors predicted fear of success (p less than .05): IPAT-Q4, IPAT-L, IPAT-E, and IPAT-O. The multiple R was .75 and the R2 was 57.78%. Interpretation of the results suggest that while this sample of Senior elite wrestlers generally exhibit extremely low fear of success scores, athletes who are tense, frustrated, apprehensive, submissive, humble, and trusting tend to be more fearful of the consequences of success than those who are relaxed, unfrustrated, self-assured, assertive, competitive, and suspecting.

Achievement↗