Chromosome aberrations induced by Eumaitenine, a sesquiterpene isolated from Maytenus boaria Mol. in cultured CHO cells.
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Animals↗
Biomedical subjects
M Silva
Publications and source records attributed to M Silva.
Automated kinetic-spectrofluorimetric method for the determination of morphine in urine.
The continuous addition of reagent technique was used with spectrofluorimetric detection for the kinetic determination of morphine in urine samples. The method thus developed is based on the oxidative dimerization of morphine to fluorescent pseudomorphine by potassium hexacyanoferrate(III) in a basic medium. The optimum pH, oxidant concentration and instrumental variables were determined. The method permits the sensitive determination of morphine over a wide concentration range (15-925 ng ml-1) with high precision (relative standard deviation approximately 2%), selectivity and sample throughput (48 samples h-1). A novel sorption--desorption procedure was used to isolate morphine from whole urine, which provided recoveries of between 85 and 100%. The results showed the usefulness of the proposed method for controlling a wide variety of medicinal problems (e.g., therapeutic, overdose and doping analysis).
Autoanalysis↗
Rotating-liquid-drop model limit tested on macroscopic drops.
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Journal Article↗
A reporter transgene indicates renal-specific induction of tumor necrosis factor (TNF) by shiga-like toxin. Possible involvement of TNF in hemolytic uremic syndrome.
We have examined the hypothesis that TNF may play a pathogenetically important role in the hemolytic uremic syndrome. Specifically, we considered the possibility that shigatoxin, which eventuates this syndrome, might induce TNF biosynthesis, and/or that TNF and shigatoxin might sensitize animals, each to the toxic effects of the other agent. Shigatoxin was found to sensitize mice to the lethal effect of LPS and to the lethal effect of TNF. On the other hand, pretreatment of animals with either TNF or LPS did not noticeably sensitize mice to the lethal effect of shigatoxin. Intraperitoneal injections of shigatoxin did not induce the production of detectable quantities of TNF in the plasma of mice. When shigatoxin was injected into transgenic mice bearing a chloramphenicol acetyltransferase (CAT) reporter gene that indicates TNF synthesis, CAT activity was induced within the kidney, but not in other tissues. We therefore conclude that shigatoxin acts to induce TNF synthesis within the kidney, and at the same time increases renal sensitivity to the toxic effects of TNF. While this mouse model does not reproduce the hemolytic uremic syndrome as it occurs in humans, it does suggest that local synthesis of TNF within the kidney may contribute to renal injury induced by shigatoxin.
Animals↗
Detection of hepatitis C virus antibody by first and second generation assays and polymerase chain reaction in patients with autoimmune chronic active hepatitis types I, II, and III.
In this study we have attempted to classify a group of North American patients with autoimmune chronic hepatitis into types I, II, and III according to the class of autoantibody present in serum, and determine the prevalence and significance of antibody to hepatitis C virus (anti-HCV). A total of 62 patients (type I, 51; type II, 3; type III, 8) were tested with first-generation enzyme-linked immunosorbent assay (ELISA)-1. Seropositive patients were assessed by second-generation recombinant immunoblot assay (RIBA)-2 and polymerase chain reaction (PCR). Our results demonstrate that 12 (19%) of the 62 patients with autoimmune hepatitis were anti-HCV ELISA-1 positive (type I, 9; type II, 1; type III, 2). Only one patient with type II autoimmune hepatitis was reactive by RIBA-2 and PCR. Eight of the 12 seropositive patients entered remission after corticosteroid therapy and seven of them became seronegative by ELISA-1. The RIBA-2 and PCR reactive patient did not respond to immunosuppressive therapy and remained seropositive. We conclude that there is a low prevalence of anti-HCV antibody in autoimmune hepatitis. Results based only on ELISA-1 anti-HCV testing can be misleading, and second-generation testing is necessary to recognize the presence of HCV infection. The fact that the only RIBA-2 reactive patient had type II autoimmune hepatitis may suggest a role for HCV infection in the pathogenesis of this condition. Nevertheless, corticosteroid therapy remains effective in those patients who are ELISA-1 seropositive, but RIBA-2 and PCR nonreactive.
Adolescent↗
[Subdural abscess. Rare complication of epidural infiltration. Apropos of a case and review of the literature].
Lumbar subdural abscesses have rarely been reported in the literature, and the complications of epidural infiltrations predominantly include arachnoiditis, meningitis and epidural abscesses. We report a case of lumbar subdural abscess confirmed by surgery, which appeared following epidural infiltration for lumbar sciatica and was examined with computed tomography and MRI. Only the latter technique alone showed the abscess, after gadolinium injection, and established its intradural site.
Adrenal Cortex Hormones↗
[Spinal subdural hematoma. Presentation of 2 cases studied with MRI].
Spinal subdural hematomas are rare. They should be considered in the diagnosis of spinal cord compression. Prompt surgical treatment, if it is performed, may lead to complete recovery of the neurological deficit. But such treatment is not always necessary because of possible spontaneous recovery. MRI appears to be the method of choice to reveal hematomas even of small size and to follow up patients after treatment.
Aged↗
[Vasoactive endothelial factors].
Endothelium-derived vasoactive factors are produced by the endothelium activated by effective stimulus, and with paracrine regulatory activity of the tone/proliferation of the vascular smooth muscle and platelet function. They are divided in two groups: endothelium-derived relaxing and contracting factors. Among the endothelium-derived relaxing factors, PG I2, EDRF (NO or other nitrous compound) and EDHF (still unidentified) have been considered Synthetized by the endothelium after stimulation by plasmatic, platelet-derived and endothelium-derived substances and mechanisms, towards the vascular smooth muscle (myorelaxing/cytostatic) and the platelets (antiaggregation). The endothelium-derived contracting factors include the EDCF1 (endothelins, 21 amino acids peptides), EDCF2 (O2-) and TxA2. Its production, induced by stimulus similar to those for relaxing factors, promotes constriction/mitogenesis of the vascular smooth muscle and platelet aggregation. Probably, endothelin-1 has indirect actions over hormonal mechanisms of cardiovascular and renal regulation. The vascular system establishes a tight regulation over the production of these endothelium-derived vasoactive factors. Its loss (usually due to alteration of endothelial responsiveness to stimulation) allows local or generalized modifications of the vascular tone. These can depend on hypertension, atherosclerosis, ischemia-reperfusion lesion, diabetes, inflammation and situations of farmacotoxicity (all developing vasoconstriction/vasospasm) or by septicemia (leading to vasodilation). This disregulation is also involved in the pathogenesis of hypertension, atherosclerosis and ischemia-reperfusion. The vascular tone regulation by endothelium also leads to systemic consequences. Essentially by decreasing cardiac, cerebral and renal blood flow it implies morphologic and functional modifications of these organs.
Animals↗
The major Thiobacillus ferrooxidans outer membrane protein forms low conductance ion channels in planar lipid bilayers.
A protein isolated and purified from the outer membrane of the acidophilic, chemolithotrophic bacterium, Thiobacillus ferrooxidans with an oligomeric molecular weight of 90,000 Da (p90) was incorporated into phosphatidylethanolamine planar lipid bilayers. The protein formed slightly anionic channels in KCl solutions, with a conductance of 25 pS in 100 mM KCl. The current-voltage relationship was linear between +/- 60 mV, and the conductance was a saturating function of the salt concentration. These channels fluctuated from a single open to closed state at low potentials, but present flickering activity at higher potentials.
Bacterial Outer Membrane Proteins↗
Prevalence of antibodies to hepatitis C virus among patients with cryptogenic chronic hepatitis and cirrhosis.
Many cases of chronic hepatitis and cirrhosis cannot be attributed to a known cause and are collectively referred to as cryptogenic chronic liver disease. We have evaluated the role of the hepatitis C virus in the pathogenesis of this condition in a retrospective serum analysis for antibody to hepatitis C virus in 129 patients with cryptogenic liver disease. Other causes of chronic hepatitis and cirrhosis were ruled out by clinical, serum biochemical and serological techniques. All 129 patients were HBcAg negative, but 28 (22%) had antibody to HBcAg. Sera were tested by radioimmunoassays using recombinant peptides for antibodies to nonstructural (C100-3 and C33c) and structural regions (C22) of HCV. Among the 129 patients, 61 (47%) had antibody to C100-3, 76 (59%) had antibody to C33c and 74 (57%) had antibody to C22. Seventy-nine (61%) were reactive with at least one and 76 (59%) were reactive with at least two HCV peptides (this is the criterion used for hepatitis C virus antibody reactivity). A proportion of patients with chronic hepatitis and cirrhosis (55 of 91; 60%) similar to that of patients without cirrhosis (21 of 38; 55%) had hepatitis C virus antibody. No significant clinical, serum biochemical or histological differences were noted between the group of patients with hepatitis C virus antibody and those without this antibody reactivity. Thus more than half the patients with cryptogenic chronic liver disease had hepatitis C virus antibody, suggesting that chronic HCV infection plays a major role in the origin of cryptogenic chronic hepatitis and cirrhosis.
Adult↗
[Evaluation of commercially available reagents for diagnosis of chagas disease in Chilean blood banks. I. Selection of reagents].
Infection with T cruzi through blood transfusion is an important public health problem in Latin America, including Chile. A study on the sensitivity, specificity and applicability of currently applied methods was performed. Ten commercially available products were tested in a panel of 180 blood specimens against the indirect immunofluorescence antibody test (IFAT) and the immunoperoxidase test (IP). Qualitative concordance ranged from 82 to 98%, sensitivity from 64 to 97% and specificity from 93 to 100%. Some commercial products had a low performance and ELISA is recommended for routine use in Chilean blood banks, given its significantly higher sensitivity and specificity when compared to the hemagglutination test. In addition, experience with the use of the ELISA test is widespread in Chile.
Animals↗
Obstetric anesthesia.
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Anesthesia, Intravenous↗
The human gene for vascular endothelial growth factor. Multiple protein forms are encoded through alternative exon splicing.
Vascular endothelial growth factor (VEGF) is an apparently endothelial cell-specific mitogen that is structurally related to platelet-derived growth factor. By Northern blot and protein analyses, we show that VEGF is produced by cultured vascular smooth muscle cells. Analysis of VEGF transcripts in these cells by polymerase chain reaction and cDNA cloning revealed three different forms of the VEGF coding region, as had been reported in HL60 cells. The three forms of the human VEGF protein chain predicted from these coding regions are 189, 165, and 121 amino acids in length. Comparison of cDNA nucleotide sequences with sequences derived from human VEGF genomic clones indicates that the VEGF gene is split among eight exons and that the various VEGF coding region forms arise from this gene by alternative splicing: the 165-amino-acid form of the protein is missing the residues encoded by exon 6, whereas the 121-amino-acid form is missing the residues encoded by exons 6 and 7. Analysis of the VEGF gene promoter region revealed a single major transcription start, which lies near a cluster of potential Sp1 factor binding sites. The promoter region also contains several potential binding sites for the transcription factors AP-1 and AP-2; consistent with the presence of these sites, Northern blot analysis demonstrated that the level of VEGF transcripts is elevated in cultured vascular smooth muscle cells after treatment with the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate.
Amino Acid Sequence↗
Immunophenotyping of lymphocytes in liver tissue of patients with chronic liver diseases by flow cytometry.
Immunological factors are important in the pathogenesis of a spectrum of hepatobiliary diseases. To characterize the nature of specific immunological responses in liver disease, we determined lymphocyte changes in liver tissue and in blood using flow cytometry. A total of 113 liver biopsy specimens was collected from patients with the following diseases: 19 chronic hepatitis B; 39 chronic non-A, non-B hepatitis; 27 alcoholic liver disease; 10 hepatic malignancy; 8 autoimmune hepatitis; 6 fatty liver and 4 primary biliary cirrhosis. The lymphocytes were isolated from the liver biopsy specimens by mechanical and enzymatic methods. The lymphocyte yield was 7,901 +/- 575 cells/mg of liver tissue. The viability of lymphocytes was 97.7% +/- 0.3%. Lymphocytes were stained with four pairs of two-color mixed fluorescein-conjugated monoclonal antibodies, including T4-T8 (CD4/CD8), T11-B1 (CD2-CD20), NKH1-T8 (CD56-CD8), IL-2R1-T11 (CD25-CD2), and the ratios were determined by an Epics Profile flow cytometer. Immunophenotyping of lymphocytes in whole blood samples was simultaneously analyzed. Variability in lymphocyte yield and different patterns of lymphocyte subsets were found in the liver biopsy specimens. The yields of lymphocytes from patients with chronic non-A, non-B and autoimmune hepatitis were highest, and the lowest yield was from patients with fatty liver. Patients with primary biliary cirrhosis, fatty liver and hepatic malignancy had relatively high ratios of CD4/CD8, CD56/CD8 and CD25/CD2; whereas patients with chronic hepatitis B, autoimmune hepatitis and non-A, non-B hepatitis had lower ratios of CD4/CD8, CD56/CD8 and CD25/CD2. No difference in lymphocyte ratios between the patients with cirrhotic and noncirrhotic alcoholic liver disease was found.(ABSTRACT TRUNCATED AT 250 WORDS)
Cell Separation↗
Simultaneous kinetic determination of epinephrine and norepinephrine by the stopped-flow technique.
A stopped-flow method for the simultaneous determination of epinephrine and norepinephrine, two catecholamines with a wide spectrum of biological activity, is proposed. The method is based on the oxidation of these compounds with 1,10-phenanthroline-iron(III) complex, which is monitored by measuring the initial rate of change of the absorbance of the ferroin formed at 510 nm. The difference in kinetic behaviour between the two species was exploited by applying a modified version of the proportional-equation method for the resolution of epinephrine-norepinephrine mixtures at the micrograms ml-1 level over the ratio range 1:10-10:1, with an error of less than 5% and an average precision of about 2.5%. The method was successfully applied to the determination of the two catecholamines in pharmaceuticals.
Epinephrine↗
Characterization of ascites present at cesarean section.
Ascites has been reported at cesarean section delivery in pregnancies complicated by preeclampsia. The frequency and composition of peritoneal fluid collections in pregnancy have not previously been described. This descriptive study evaluated presence, composition, and characteristics of ascites present in 41 women undergoing cesarean delivery for a variety of indications. Twenty-six women had sufficient quantities of peritoneal fluid to be collected immediately after opening the parietal peritoneum. The protein content of the fluid was analyzed. The presence of fluid was not related to labor preceding delivery. All preeclamptic women in this study had fluid obtainable at cesarean section. Large volume ascites was noted in five preeclamptic women whose fluid had low total protein content, consistent with a transudate. Intrahepatic portal hypertension is one possible explanation of the large volume and low protein ascites in some preeclamptic patients. Ultrasonic measurements in preeclamptic patients and normotensive pregnant women showed uniform portal vein diameters and Doppler-derived portal vein flow velocities in all patients. The physiology of peritoneal fluid production in complicated and normal pregnancy remains unexplained.
Adult↗
Inhibition of hematopoiesis in vitro with the Lym-1 monoclonal antibody.
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Antibodies, Monoclonal↗
Expression of a negative regulator of human erythropoiesis by fluidized lymphocyte plasma membranes.
Regulation of hematopoiesis by paracrine molecules occurs in vitro. In some cases, hematopoietic paracrine factors have been localized to the plasma membrane of accessory cells. We have purified a unique integral membrane glycoprotein from normal human B cells that functions in vitro as a paracrine factor whose activity is directed toward erythroid progenitor cells. This factor is also spontaneously exfoliated from the cell surface as a component of extracellular vesicles. Analysis of the lipid and protein compositions and membrane lipid order of these extracellular vesicles reveals them to be biochemically distinct and more fluid than their parent membranes. Evidence in nonhematopoietic culture systems indicates that cell membrane function may be altered by modifying membrane fluidity. In an effort to accelerate growth factor release, plasma membranes of B cells were fluidized by incubation with an emulsion of Liposin II, phosphatidylcholine, and phosphatidylethanolamine. Fluidity assessed by steady-state fluorescence polarization was reduced in lipid-treated cells. Exfoliation was 3-4-fold higher from lipid-treated cells relative to untreated cells. Unexpectedly, a negative signal for burst-forming unit-erythroid (BFU-E) proliferation was expressed in membranes, in shed extracellular vesicles, and in supernatants of medium conditioned by the fluidized cells. Purification of the inhibitor is under way. The data are consistent with the hypothesis that accessory cell plasma membranes may positively or negatively regulate erythroid differentiation, depending upon the exchange of cholesterol and phospholipids between plasma membrane and ambient lipid pools.
B-Lymphocytes↗