983 resultados para Microscopic observation drug susceptibility assay (MODS)


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RESUMO: Desde 1976 que as Forças Armadas desenvolvem acções de prevenção do consumo de drogas e álcool. Na década de 80 foi criada capacidade laboratorial e deu-se início a um programa de rastreios toxicológicos. No quinquénio 2001 a 2005, as proporções de resultados positivos, associando todos os tipos de rastreio, variaram entre 3,7% e 1,5%. De Outubro de 2006 a Julho de 2007 realizou-se um estudo analítico transversal, para estimar a prevalência do consumo de drogas (canabinóides, opiáceos, cocaína e anfetaminas) num dos Ramos das Forças Armadas, com base nos despistes realizados pelo seu laboratório. Foi utilizada uma amostra aleatória simples de 1039 militares, profissionais (QP) e contratados (RC), no activo e de ambos os sexos. Desde a nomeação dos militares a rastrear, passando pela cadeia de custódia das amostras até à obtenção do resultado foi utilizado apoio informático específico. O processo de pesquisa utilizou duas técnicas de triagem por imunoensaio e tecnologia de confirmação por GC/MS, de acordo com as recomendações europeias, permitindo estabelecer uma metodologia standard para organizações e empresas. A prevalência estimada, de consumidores de droga, foi de 3,8/1.000, para um erro de 0,37%. O número de casos registado (4) não permitiu a utilização de testes estatísticos que conduzissem à identificação de características determinantes da positividade, mas não deixou de revelar aspectos inesperados. A observação de séries de casos e a realização regular de estudos epidemiológicos, que ajudem a redefinir grupos alvo e a perceber a dimensão, as determinantes e as consequências do consumo de drogas é sugerida, em conclusão.--------------------------------------- RÉSUMÉ: Depuis 1976, les Forces Armées mettent au point des mesures visant à prévenir la consommation de drogues et d'alcool. En 1980, fut créé capacité laboratoriel et ont ensuite commencé un programme de dépistage toxicologique. Au cours des cinq années allant de 2001 à 2005, les proportions de consommateurs, impliquant tous les types de dépistage, allaient de 3,7% à 1,5 %. D'octobre 2006 à juillet 2007, une étude analytique transversale a été organisée pour évaluer la prévalence de l’usage de drogues (cannabis, opiacés, cocaïne et amphétamines) dans une branche de les Forces Armées, basée sur les dépistages faites par un laboratoire militaire, à l'aide d'un échantillon aléatoire de 1039 militaires, professionnels (QP) et sous contract (RC), à l’actif et des deux sexes. Tout au long du procès, de la nomination des donneurs, en passant par la chaine de garde des échantillons, jusqu’à obtention du résultat, il fut employé un appui informatique sécurisé. Le processus de recherche employa deux techniques de tri par imunoessay et la technologie de confirmation GC/MS, selon les recommandations européennes, permettant d'établir une méthodologie standard pour les organisations et les entreprises. La prévalence estimée fut de 3,8/1.000 pour une marge d’erreur de 0,37%. Le nombre de cas enregistrés (4) n'autorise pas l'utilisation de testes statistiques de menant à l'identification de caractéristiques déterminant de la positivité, mais il permet à révéler des aspects inattendus. L'observation de séries de cas et la tenue régulière d’études épidémiologiques, qui contribuent à redéfinir les groupes cibles et de comprendre l'ampleur, les déterminants et les conséquences de l'usage de drogues, est suggéré, en fin de compte.--------------------------------------- ABSTRACT: Since 1976, the Armed Forces, have been developing measures to prevent the use of drugs and alcohol. In 1980, was created laboratory facility which then started a program of toxicological screenings. In the five years running from 2001 to 2005, the proportions of consumers, involving all types of screening, ranged from 3,7% to 1,5%. From October 2006 to July 2007, a cross-sectional study was held to estimate the prevalence of drug use (cannabinoids, opiates, cocaine and amphetamines) in one branch of the Portuguese Armed Forces, based on laboratory screenings, using a random sample of 1039 military, professional (QP) and enlisted (RC), active-duty and of both sexes. Specific computer support was used all the way, from the appointment, including the chain of custody of samples, to the obtaining of the result. The process of search used two techniques for sorting by immunoassay and confirmation technology GC/MS, according to European recommendations, allowing to establish a standard methodology for organizations and companies. The estimated prevalence of drug users was 3.8/1.000 for a 0.37% error (95% confidence interval). The number of cases registered (4) does not permit use of statistical testing leading to the identification of characteristics weighing in the establishing to extrapolate for the population, but it allows revealing unexpected aspects. The observation of series of cases and the regular holding of epidemiological studies, which help redefine target groups and to understand the extent, the determinants and consequences of drug use, is suggested, in conclusion.

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Extended-spectrum β-lactamases (ESBLs) prevalence was studied in the north of Portugal, among 193 clinical isolates belonging to citizens in a district in the boundaries between this country and Spain from a total of 7529 clinical strains. In the present study we recovered some members of Enterobacteriaceae family, producing ESBL enzymes, including Escherichia coli (67.9%), Klebsiella pneumoniae (30.6%), Klebsiella oxytoca (0.5%), Enterobacter aerogenes (0.5%), and Citrobacter freundii (0.5%). β-lactamases genes blaTEM, blaSHV, and blaCTX-M were screened by polymerase chain reaction (PCR) and sequencing approaches. TEM enzymes were among the most prevalent types (40.9%) followed by CTX-M (37.3%) and SHV (23.3%). Among our sample of 193 ESBL-producing strains 99.0% were resistant to the fourth-generation cephalosporin cefepime. Of the 193 isolates 81.3% presented transferable plasmids harboring genes. Clonal studies were performed by PCR for the enterobacterial repetitive intragenic consensus (ERIC) sequences. This study reports a high diversity of genetic patterns. Ten clusters were found for E. coli isolates and five clusters for K. pneumoniae strains by means of ERIC analysis. In conclusion, in this country, the most prevalent type is still the TEM-type, but CTX-M is growing rapidly.

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In today’s healthcare paradigm, optimal sedation during anesthesia plays an important role both in patient welfare and in the socio-economic context. For the closed-loop control of general anesthesia, two drugs have proven to have stable, rapid onset times: propofol and remifentanil. These drugs are related to their effect in the bispectral index, a measure of EEG signal. In this paper wavelet time–frequency analysis is used to extract useful information from the clinical signals, since they are time-varying and mark important changes in patient’s response to drug dose. Model based predictive control algorithms are employed to regulate the depth of sedation by manipulating these two drugs. The results of identification from real data and the simulation of the closed loop control performance suggest that the proposed approach can bring an improvement of 9% in overall robustness and may be suitable for clinical practice.

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Several reports have related Legionella pneumophila with pneumonia in renal transplant patients, however this association has not been systematically documented in Brazil. Therefore this paper reports the incidence, by serologycal assays, of Legionella pneumophila serogroup 1 in these patients during a five year period. For this purpose sera from blood samples of 70 hospitalized patients with pneumonia from the Renal Transplant Unit of Hospital das Clinicas, FMUSP collected at the acute and convalescent phase of infection were submitted to indirect immunofluorescence assay (IFA) to demonstrate anti-Legionella pneumophila serogroup 1 antibodies. Of these 70 patients studied during the period of 1988 to 1993,18 (25.71 %) had significant rises in specific antibody titers for Legionella pneumophila serogroup 1. Incidence was interrupted following Hospital water decontamination procedures, with recurrence of infections after treatment interruption. In this study, the high susceptibility (25.71%) of immunodepressed renal transplant patients to Legionella pneumophila serogroup 1 nosocomial infections is documented. The importance of the implementation and maintenance of water decontamination measures for prophylaxis of the infection is also clearly evident.

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The localization of magma melting areas at the lithosphere bottom in extensional volcanic domains is poorly understood. Large polygenetic volcanoes of long duration and their associated magma chambers suggest that melting at depth may be focused at specific points within the mantle. To validate the hypothesis that the magma feeding a mafic crust, comes from permanent localized crustal reservoirs, it is necessary to map the fossilized magma flow within the crustal planar intrusions. Using the AMS, we obtain magmatic flow vectors from 34 alkaline basaltic dykes from São Jorge, São Miguel and Santa Maria islands in the Azores Archipelago, a hot-spot related triple junction. The dykes contain titanomagnetite showing a wide spectrum of solid solution ranging from Ti-rich to Ti-poor compositions with vestiges of maghemitization. Most of the dykes exhibit a normal magnetic fabric. The orientation of the magnetic lineation k1 axis is more variable than that of the k3 axis, which is generally well grouped. The dykes of São Jorge and São Miguel show a predominance of subhorizontal magmatic flows. In Santa Maria the deduced flow pattern is less systematic changing from subhorizontal in the southern part of the island to oblique in north. These results suggest that the ascent of magma beneath the islands of Azores is predominantly over localized melting sources and then collected within shallow magma chambers. According to this concept, dykes in the upper levels of the crust propagate laterally away from these magma chambers thus feeding the lava flows observed at the surface.

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A new serological test, the gelatin particle agglutination test (GPAT), was used for the serodiagnosis of schistosomiasis mansoni. This technique showed the sensitivity (90.6%) and specificity (97.8%) close to those of enzyme-linked immunosorbent assay. The GPAT can be easily and rapidly performed without specialized equipment, by using lyophilized antigen-coated gelatin particles. The test also seems to be useful for mass screening of Schistosoma infection in field conditions.

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B. tenagophila snails from Ouro Branco, MG, showed positivity for S. mansoni, with infection rates of 5%, 10%, (SJ strain), and 1% (LE strain) using a pool of miracidia. The mollusks were found to be susceptive from the 3rd generation reared in laboratory onwards. The B. tenagophila (OB, MG) when individually exposed to 10 miracidia, showed infection rate of 2% for LE strain. B. glabrata snails from Gagé, MG, showed a positivity rate of 58% for S. mansoni (LE strain), under experimental conditions. The B. tenagophila from Cabo Frio, RJ and B. glabrata from Belo Horizonte, MG used as a control for SJ strain showed infection rates of 47% - 85% and 36% respectivily. For the LE strain, B. glabrata (BH, MG) used as control showed infection rate of 40% - 75%.

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A dot-enzyme-linked immunosorbent assay (Dot-ELISA) for pneumococcal antigen detection was standardized in view of the need for a rapid and accurate immunodiagnosis of acute pneumococcal pneumonia. A total of 442 pleural fluid effusion samples (PFES) from children with clinical and laboratory diagnoses of acute bacterial pneumonia, plus 38 control PFES from tuberculosis patients and 20 negative control serum samples from healthy children were evaluated by Dot-ELISA. The samples were previously treated with 0.1 M EDTA pH 7.5 at 90°C for 10 min and dotted on nitrocellulose membrane. Pneumococcal omniserum diluted at 1:200 was employed in this assay for antigen detection. When compared with standard bacterial culture, counterimmunoelectrophoresis and latex agglutination techniques, the Dot-ELISA results showed relative indices of 0.940 to sensitivity, 0.830 to specificity and 0.760 to agreement. Pneumococcal omniserum proved to be an optimal polyvalent antiserum for the detection of pneumococcal antigen by Dot-ELISA. Dot-ELISA proved to be a practical alternative technique for the diagnosis of pneumococcal pneumonia.

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The sensitivity and specificity of an enzyme-linked immunosorbent assay (ELISA) for the detection of circulating antigens from toxic components of Tityus serrulatus scorpion venom was determined in patients stung by T. serrulatus before antivenom administration. Thirty-seven patients were classified as mild cases and 19 as moderate or severe cases. The control absorbance in the venom assay was provided by serum samples from 100 individuals of same socioeconomic group and geographical area who had never been stung by scorpions or treated with horse antisera. The negative cutoff value (mean + 2 SD) corresponded to a venom concentration of 4.8 ng/ml. Three out of the 100 normal sera were positive, resulting in a specificity of 97%. The sensitivity of the ELISA when all cases of scorpion sting were included was 39.3%. When mild cases were excluded, the sensitivity increased to 94.7%. This study showed that this ELISA can be used for the detection of circulating venom toxic antigens in patients with systemic manifestations following. T. serrulatus sting but cannot be used for clinical studies in mild cases of envenoming since the test does not discriminate mild cases from control patients.

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The aim of this study was to determine the value of blood culture as a parameter of treatment effectiveness in experimental histoplasmosis. A total of thirty five hamsters, weighing approximately 120g, were inoculated intracardiacly with 0.1 ml of a suspension containing 4 x 10(7) cells/ml of the yeast phase of H. capsulatum. Treatments were started one week after the infection and lasted for 3 weeks. The azoles, (itraconazole, saperconazole and fluconazole) were administered once a day by gavage, at a dose of 8 mg/kg; Amphotericin B was given intraperitonealy every other day at a dose of 6mg/kg. Blood samples (1 ml) were obtained by heart punction from the 4th day after infection and were seeded in Sabouraud honey-agar and BHI-agar. The hamsters that survived were killed one week after treatment completion and the following criteria were considered for treatment evaluation: 1) rate of spontaneous death, at the end of the experience; 2) microscopic examination of Giemsa smears from liver and spleen and 3) determination of CFU in spleen cultures. Amphotericin B was the most effective drug, with negative blood cultures at day 20, negative spleen cultures in all cases and all the animals survived until the end of the study. Fluconazole was the less effective drug, blood cultures were positive during the whole experience, spleen cultures showed a similar average of CFU when compared with the control animals and 42.8% of these animals died. Saperconazole and itraconazole showed a similar activity, with survival of all hamsters and negative blood cultures at 23 and 26 days respectively. Blood culture seems to be valuable parameter for treatments' evaluation in experimental histoplasmosis of the hamster.

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A Dot enzyme-linked immunosorbent assay (Dot-ELISA) was standardized and evaluated for the serodiagnosis of human toxoplasmosis. Out of 538 serum samples tested by the immunofluorescence test for toxoplasmosis (IFAT-IgG) as reference test, 183 (34%) were positive at cut off 1:16 and 192 (36%) were positive for Dot-ELISA-IgG at cut-off 1:256. For Dot-ELISA, co-positivity was 0.94, co-negativity 0.94 and concordance 0.88 in relation to IFAT-IgG. These results suggest the usefulness of Dot-ELISA (cut-off titer of 1:256) for the serodiagnosis of human toxoplasmosis. The main advantage of this technique is simplicity, positive test can be visually identified (colored precipitate). It does not require a special equipment and it can be used as a qualitative test to screen large numbers of samples or as a quantitative assay to determine end-point titration of individual sera.

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An immunoprecipitation technique, ELIEDA (enzyme-linked-immuno-electro-diffusion assay), was evaluated for the diagnosis of Schistosoma mansoni infection with low worm burden. One hundred of serum samples from patients excreting less than 600 eggs per gram of feces (epg), with unrelated diseases and clinically healthy subjects were studied. In patients with egg counts higher than 200 epg, the sensitivities of IgM and IgG ELIEDA were 1.000 and 0.923, respectively, not differing from other Serologic techniques, such as indirect hemaglutination (IHAT), immunofluorescence (IFT) tests and immuno-electrodiffusion assay (IEDA). However in patients with low egg counts (< 100 epg), the IgG ELIEDA provided better results (0.821) than IgM ELIEDA (0.679), showing sensitivity that did not differ from that of IgG IFT (0.929), but lower than that of IgM IFT (0.964). However, its sensivity was higher than that found with IHAT (0.607) and IEDA (0.536). The specificity of IgG ELIEDA was comparable to that of other techniques. The data indicate that IgG ELIEDA might be useful for the diagnosis of slight S. mansoni infections, and the cellulose acetate membrane strips can be stored for further retrospective studies.

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A simple method of rubella antigen production by treatment with sodium desoxycholate for use in enzyme immunoassay (IMT-ELISA) is presented. When this assay was compared with a commercial test (Enzygnost-Rubella, Behring), in the study of 108 sera and 118 filter paper blood samples, 96.9% (219/226) overall agreement and correlation coefficient of 0.90 between absorbances were observed. Seven samples showed discordant results, negative by the commercial kit and positive by our test. Four of those 7 samples were available, being 3 positive by HI.

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We studied the susceptibility to Leishmania (Viannia) panamensis in strains of mice. The C57BL/6 strain was resistant and showed self-controlled lesion at the injected foot pad. The BALB/c and DBA/2J strains were susceptible and showed a foot swelling that started day 20 post-infection and progressed to a tumour-like lesion in later period of observation. The CBA/HJ strain was found to be of intermediary resistance. In contrast to other known cutaneous leishmaniasis in mice, the lesion in L. (V.) panamensis-infected mice was restricted to the inoculation site in the skin. In addition, we studied the development of cellular response and antibodies against Leishmania antigen in BALB/c and C57BL/6 strains. The proliferative response of lymph node cells against L. (V.) panamensis antigen was biphasic in both strains. An initial response was seen on day 20, followed by a refractory period between 40 and 80 days and a second response around fourth month post-infection. The response in the latter period was higher in C57BL/6 strain than in BALB/c strain. BALB/c strain presented much higher anti-Leishmania antibody level than C57BL/6 strain. The model and the correlation of immunological variables and the course of the infection are discussed.

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M. tuberculosis-positive cultures were obtained from 228 patients seen in our service and drug sensitivity assays were carried out from January 1992 to December 1994. A survey of the medical records of these patients showed resistance to one or more drugs in 47 (20.6%), 25 of whom (10.9%), who reported previous treatment, were considered to have acquired resistance. Among the antecedents investigated, only previous treatment and alcoholism were the factors independently associated with the occurrence of resistance. The survival of patients with resistant strains was lower than that of patients attacked by non-resistant M. tuberculosis. We conclude that in the present series M. tuberculosis resistance to tuberculostatic agents was predominantly of the acquired type.