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Biomedical subjects

E Gomes

Publications and source records attributed to E Gomes.

At least 19 recordsLinked to original sources

A method for multiple sequential analyses of macrophage functions using a small single cell sample.

Microbial pathogens such as bacillus Calmette-Guérin (BCG) induce the activation of macrophages. Activated macrophages can be characterized by the increased production of reactive oxygen and nitrogen metabolites, generated via NADPH oxidase and inducible nitric oxide synthase, respectively, and by the increased expression of major histocompatibility complex class II molecules (MHC II). Multiple microassays have been developed to measure these parameters. Usually each assay requires 2-5 x 10(5) cells per well. In some experimental conditions the number of cells is the limiting factor for the phenotypic characterization of macrophages. Here we describe a method whereby this limitation can be circumvented. Using a single 96-well microassay and a very small number of peritoneal cells obtained from C3H/HePas mice, containing as little as <=2 x 10(5) macrophages per well, we determined sequentially the oxidative burst (H2O2), nitric oxide production and MHC II (IAk) expression of BCG-activated macrophages. More specifically, with 100 micro l of cell suspension it was possible to quantify H2O2 release and nitric oxide production after 1 and 48 h, respectively, and IAk expression after 48 h of cell culture. In addition, this microassay is easy to perform, highly reproducible and more economical.

Animals↗

Purification and characterization of two beta-glucosidases from the thermophilic fungus Thermoascus aurantiacus.

beta-Glucosidase from the fungus Thermoascus aurantiacus grown on semi-solid fermentation medium (using ground corncob as substrate) was partially purified in 5 steps--ultrafiltration, ethanol precipitation, gel filtration and 2 anion exchange chromatography runs, and characterized. After the first anion exchange chromatography, beta-glucosidase activity was eluted in 3 peaks (Gl-1, Gl-2, Gl-3). Only the Gl-2 and Gl-3 fractions were adsorbed on the gel matrix. Gl-2 and Gl-3 exhibited optimum pH at 4.5 and 4.0, respectively. The temperature optimum of both glucosidases was at 75-80 degrees C. The pH stability of Gl-2 (4.0-9.0) was higher than Gl-3 (5.5-8.5); both enzyme activities showed similar patterns of thermostability. Under conditions of denaturing gel chromatography the molar mass of Gl-2 and Gl-3 was 175 and 157 kDa, respectively. Using 4-nitrophenyl beta-D-glucopyranoside as substrate, Km values of 1.17 +/- 0.35 and 1.38 +/- 0.86 mmol/L were determined for Gl-2 and Gl-3, respectively. Both enzymes were inhibited by Ag+ and stimulated by Ca2+.

Ascomycota↗

Production, characterization and properties of polysaccharide depolymerizing enzymes from a strain of Curvularia inaequalis.

Xylanase, beta-glucosidase, beta-xylosidase, endoglucanase and polygalacturonase production from Curvularia inaequalis was carried out by means of solid-state and submerged fermentation using different carbon sources. beta-Glucosidase, beta-xylosidase, polygalacturonase and xylanase produced by the microorganisms were characterized. beta-Glucosidase presented optimum activity at pH 5.5 whereas xylanase, polygalacturonase and beta-xylosidase activities were optimal at pH 5.0. Maximal activity of beta-glucosidase was determined at 60 degrees C, beta-xylosidase at 70 degrees C, and polygalacturonase and xylanase at 55 degrees C. These enzymes were stable at acidic to neutral pH and at 40-45 degrees C. The crude enzyme solution was studied for the hydrolysis of agricultural residues.

Ascomycota↗

Suppression of asthma-like responses in different mouse strains by oral tolerance.

In this study we examined the effect of oral antigen (Ag) administration on the development of experimental asthma in different mouse strains. We selected BALB/c, BP2, CBA/Ca interleukin (IL)-5 transgenic, and BALB/c T-cell receptor-delta-deficient mouse strains because they exhibit different aspects of the asthma syndrome. Mice exposed to 1% ovalbumin (OVA), dissolved in the drinking water for 5 consecutive days, became unresponsive to subsequent immunogenic OVA challenges. This regimen of OVA administration induced Ag-specific unresponsiveness in all mouse strains tested, including gammadelta-deficient mice that are said to be resistant to tolerance induction. The Ag-specific unresponsiveness was characterized by reduced (almost absent) airway eosinophilic inflammation, airway hyperreactivity, and mucus production; also by low levels of T helper (Th) 2-type cytokines in bronchoalveolar lavage fluid, and decreased immunoglobulin (Ig) G1 and IgE OVA-specific antibody production. The unresponsive state was not associated with increased levels of the suppressive cytokines IL-10 and transforming growth factor (TGF)-beta or with immune deviation toward the Th1 pathway due to increased levels of interferon-gamma and IL-12. Moreover, treatment with anti- TGF-beta antibodies did not abrogate oral tolerance. Oral Ag administration was quite effective in suppressing the development of key features of asthma when initiated after primary immunization (Day 0) or after booster (Day 7), but not after challenge (Day 14) when it increased allergic responses. Collectively, our findings show for the first time the beneficial and detrimental effects of oral Ag administration on the development of experimental asthma.

Administration, Inhalation↗

[Can blood homocysteine explain the family history of vascular diseases?].

INTRODUCTION AND AIMS: Family history of vascular disease is an important risk factor for vascular disease, independent of conventional risk factors. Homocysteinemia, a newly defined risk factor, is caused by genetics, such as cystathionine beta synthase deficiencies, and metabolic deficiencies. With the present work we intend to study the influence of family history of vascular disease in homocysteinemia. METHODS: We studied 204 normal persons (153 males), average age 38.7 +/- 10.9 years, in terms of family history of vascular disease (death due to myocardial infarction or a stroke), conventional risk factors, routine laboratory tests, fasting homocysteinemia and after oral methionine loading (0.1 g/Kg body weight). We compared laboratory results, conventional risk factors and homocysteinemia levels in persons with and without a family history of vascular disease. We performed covariance analysis to evaluate, in a multivariate model, factors that were related to basal or after methionine loading homocysteinemia. RESULTS: 35% of persons presented a family history of vascular disease (FHVD). Persons with FHVD presented higher age (45.6 +/- 8.9 versus 35.0 +/- 10.1, p < 0.001), and higher prevalence of hypertension (p = 0.002), dyslipidemia (p = 0.001), obesity (p = 0.03), and physical inactivity (p = 0.03). They presented a tendency, without statistical significance, to have a higher prevalence of diabetes and of hyperhomocysteinemia, and to present higher levels of basal and afterload homocysteinemia. Performing covariance analysis, basal homocysteinemia did not present any relation to FHVD. After methionine load homocysteinemia was strongly influenced by basal homocysteinemia (p = 0.0000), and significantly related to FHVD (p = 0.039). CONCLUSIONS: Homocysteinemia cannot explain most of the risk of family history of vascular disease, not explained by conventional risk factors. The only significant relationship between homocysteinemia and FHVD was observed with afterload homocysteinemia in the multivariate model. FHVD is clearly related to conventional risk factors.

Adult↗

Hepatic glucokinase is induced by dietary carbohydrates in rainbow trout, gilthead seabream, and common carp.

Glucokinase (GK) plays a central role in glucose homeostasis in mammals. The absence of an inducible GK has been suggested to explain the poor utilization of dietary carbohydrates in rainbow trout. In this context, we analyzed GK expression in three fish species (rainbow trout, gilthead seabream, and common carp) known to differ in regard to their dietary carbohydrate tolerance. Fish were fed for 10 wk with either a diet containing a high level of digestible starch (>20%) or a diet totally deprived of starch. Our data demonstrate an induction of GK gene expression and GK activity by dietary carbohydrates in all three species. These studies strongly suggest that low dietary carbohydrate utilization in rainbow trout is not due to the absence of inducible hepatic GK as previously suggested. Interestingly, we also observed a significantly lower GK expression in common carp (a glucose-tolerant fish) than in rainbow trout and gilthead seabream, which are generally considered as glucose intolerant. These data suggest that other biochemical mechanisms are implicated in the inability of rainbow trout and gilthead seabream to control blood glucose closely.

Animals↗

Increased replication of T-cell-tropic HIV strains and CXC-chemokine receptor-4 induction in T cells treated with macrophage inflammatory protein (MIP)-1alpha, MIP-1beta and RANTES beta-chemokines.

OBJECTIVE AND DESIGN: To study, in T-lymphoid cells, the effects of macrophage inflammatory protein (MIP)-1alpha, MIP-1beta and RANTES beta-chemokines on the replication of T-cell-tropic HIV-1 strains, since it has been reported that beta-chemokines interfere with the replication of macrophage-tropic HIV-1 strains, but not T-cell-tropic strains. METHODS: Freshly phytohaemagglutinin (PHA)-activated peripheral blood lymphocytes (PBL) and cultured PHA-activated T cells from healthy volunteers, as well as the C8166 T-cell line, were treated overnight with beta-chemokines before infection with T-cell-tropic HIV-1 isolates, or human T-lymphotropic virus type IIIB. HIV replication was followed by detecting the production of infectious particles, p24 antigen, and viral sequences. CXC-chemokine receptor (CXCR)-4 expression was followed by detection and quantification of specific transcripts. RESULTS: Pretreatment of T cells with MIP-1alpha, MIP-1beta and RANTES affected T-cell-tropic strains, increased the replication of HIV-1beta and HIV-1RPdT strains dose-dependently, as well as virus absorption and provirus DNA accumulation. These findings were associated with increased accumulation of CXCR-4 transcripts, and mediated by the protein tyrosine kinase signalling. Moreover, beta-chemokines stimulated PBL proliferation. CONCLUSIONS: Beta-chemokines increase the adsorption and replication of at least some T-cell-tropic HIV-1 strains, and this is related to stimulated expression of the CXCR-4 coreceptor.

Cell Division↗

Prevention of lung eosinophilic inflammation by oral tolerance.

Airway inflammation plays a major role in human asthma. Increasing evidence points to a close correlation between eosinophil infiltration and allergic lung disease. A new murine model of eosinophilic lung inflammation has recently been developed; it consists of immunizing mice with small fragments of solidified hen egg white implanted (EWI) into the subcutaneous tissue. In this model, which is further characterized here, mice challenged with ovalbumin (OVA) present an intense and persistent lung eosinophilia, as well as histopathological findings that resemble human asthma. In the present work, the effect of oral tolerance on the development of allergic lung inflammation in B6 mice immunized with antigen plus adjuvant or with EWI is investigated. It was found that in mice rendered orally tolerant by previous exposure to antigen in the drinking water, the T-helper type 2 cell (Th2)-associated allergic responses in both protocols of immunization were almost completely abolished. The allergic responses were assessed by pulmonary and bone marrow eosinophilia, lung histopathology and antigen-specific IgE and IgG1 production. These findings provide the first indication that Th2-associated lung pathology can be prevented by oral tolerance.

Administration, Oral↗

Antimicrobial activity of Terminalia macroptera root.

Terminalia macroptera Guill et Perr. (Combretaceae) is a medicinal plant used in Guinea-Bissau and other West African countries to treat infectious diseases. The ethanol extract from T. macroptera decorticated root and their liquid-liquid partition fractions, were screened for antimicrobial activity, by the twofold serial microdilution assay against seven reference bacterial strains and against Candida albicans. The extract and fractions showed some activity against at least one of the test microorganisms. The best results were obtained against Shigella dysenteriae and Vibrio cholerae. The minimum inhibitory concentrations (MIC) of T. macroptera ethanol extract were also determined for about 100 clinical strains of Campylobacter sp., Escherichia coli, Salmonella sp., Shigella sp. and Vibrio cholerae. The ethanol extract activity against Campylobacter strains is similar to co-trimoxazole, higher than sulfamethoxazole but lower than tetracycline, erythromycin, ampicillin and streptomycin. Ellagitannins are the major compounds in the extract and active fractions. The obtained results suggest a potential importance of this medicinal plant in the treatment of enteric diseases, particularly in Campylobacter infections.

Anti-Bacterial Agents↗

Outcome of cardiopulmonary resuscitation in a Portuguese university hospital.

Portugal has many weak links in the so-called 'chain of survival' both in the pre-hospital and in-hospital setting. Apart from evaluating the performance of a newly implemented in-hospital cardiac arrest system, we assessed the correlation between different clinical variables and outcome after cardiopulmonary resuscitation (CPR). All resuscitation attempts during 1995 were registered using the form recommended by the European Resuscitation Council. One hundred and twenty-five patients either collapsing out-of-hospital (42; 33.6%) or having in-hospital cardiac arrest (83; 66.4%) were included in the study. The number of patients who experienced recovery of spontaneous circulation was 10 (23.8%) in the out-of-hospital group and 35 (42.2%) in the in-hospital one. Of all out-of-hospital patients, 11.9% were discharged from hospital compared with 22.8% of the in-hospital group. Asystole was the initial rhythm in a significant number of arrests. The average call-response interval of the in-hospital cardiac arrest team was under 3 min. The results from this series concur with other reported series. Although good standards of care were achieved, we are aware that this was only an isolated step in the implementation of the 'chain of survival' in our country. The authors conclude that there is an urgent need for a nationwide programme that improves the standards of care for these patients.

Adult↗

Antimicrobial activity of Guinea-Bissau traditional remedies.

The ethanolic extracts of twelve plants selected through ethnomedical survey in Guinea-Bissau were investigated for their in vitro antimicrobial properties over ten bacteria and Candida albicans, using agar diffusion and dilution methods. All the tested extracts showed some activity against at least one of the bacteria. Most of the extracts (79%) showed activity against Staphylococcus aureus and only one (Cryptolepis sanguinolenta) against Escherichia coli. Cryptolepis sanguinolenta and Terminalia macroptera root extracts showed some activity against Candida albicans as well as showing an interesting profile of activity against most of the enteropathogen microorganisms. Inhibition zones against Staphylococcus aureus were localised on extract chromatograms by bioautographic techniques.

Anti-Bacterial Agents↗

Wet versus dry enamel ablation by Er:YAG laser.

The purpose of this study was to observe tooth structure and pulpal temperature changes in extracted human teeth subjected to a pulsed Er:YAG (2.94 microns) laser. Two teeth were irradiated while dry and three teeth while moistened by a fine water mist. When the dry teeth were irradiated, there was minimal enamel ablation. SEM of the resulting surface showed rounded fragments of enamel rods, enamel melting, cracks, and smooth-edged voids. Intrapulpal temperature measured by thermal sensor rose more than 27 degrees C. When the laser application on the teeth was pulsed with a constant fine water mist, enamel and dentin were efficiently ablated. SEM of the resulting surfaces showed fissures and conical craters with sharp enamel projections remaining. Intrapulpal temperatures rose an average of 4 degrees C. These results indicate that pulsed Er:YAG (2.94 microns) used with a water mist removes enamel and dentin without producing significant pulpal temperature changes.

Aerosols↗

Community-based evaluation survey of immunizations in Burkina Faso.

A cluster sample survey was conducted in January 1989 in 3 provinces of Burkina Faso to evaluate an immunization programme (based on two contacts, providing inactivated poliomyelitis vaccine plus DPT) that had been launched in 1982-84. The objectives were to estimate neonatal tetanus (NNT) mortality and poliomyelitis prevalence in the study area. The target population (using the same sample of households comprised 2107 live infants born during the preceding year for the NNT survey, and 17,154 children aged 0-9 years for the poliomyelitis survey. The NNT mortality rate was 3.3 per 1000 live births, and the poliomyelitis prevalence rate was 2.8 per 1000 children aged 5-9 years. Dates of onset of poliomyelitis cases among children aged 0-9 years and the numbers of children at risk during the 10-year recall period, reconstituted with demographic indicators taken from standardized life-tables, were used to calculate the incidence rates of poliomyelitis. These rates could be compared in the 5-year period preceding the survey, and showed a decreasing trend consistent with routine surveillance data.

Burkina Faso↗