The use of oligonucleotide probes for meningococcal serotype characterization

In the present study we examine the potential use of oligonucleotide probes to characterize Neisseria meningitidis serotypes without the use of monoclonal antibodies (MAbs). Antigenic diversity on PorB protein forms the bases of serotyping method. However, the current panel of MAbs underestimated, by at least 50% the PorB variability, presumably because reagents for several PorB variable regions (VRs) are lacking, or because a number of VR variants are not recognized by serotype-defining MAbs12. We analyzed the use of oligonucleotide probes to characterize serotype 10 and serotype 19 of N. meningitidis. The porB gene sequence for the prototype strain of serotype 10 was determined, aligned with 7 other porB sequences from different serotypes, and analysis of individual VRs were performed. The results of DNA probes 21U (VR1-A) and 615U (VR3-B) used against 72 N. meningitidis strains confirm that VR1 type A and VR3 type B encode epitopes for serotype-defined MAbs 19 and 10, respectively. The use of probes for characterizing serotypes possible can type 100% of the PorB VR diversity. It is a simple and rapid method specially useful for analysis of large number of samples.

Genetic relationships among serogroup B: serotype 4 Neisseria meningitidis strains

We compared the results obtained by serotyping of PorB epitopes using an expanded panel of monoclonal antibodies (mAb) including mAb 7 and mAb 10, with results obtained by RFLP of rRNA genes (ribotyping). The purpose of this study was to assess the correlation between phenotypic- and genotypic- methods for typing N. meningitidis. The ribotypes obtained using ClaI or EcoRV endonucleases grouped the strains in seven and two different patterns, respectively. This additional characterization of PorB epitopes improved the correlation between these two methods of typing N. meningitidis.

Histopathology, serology and cultures in the diagnosis of cryptococcosis

Cryptococcosis is one of the most common opportunistic fungal infections in patients with acquired immunodeficiency syndrome (AIDS). We report 13 cases of cryptococcal infection based on histopathology, serology and cultures. Epidemiological analysis, histochemical techniques of hematoxilin and eosin (HE) and Grocot's silver (GMS), as well special histochemical techniques such as Mayer's mucicarmine (MM) and Fontana-Masson (FM), cryptococcal antigen test (CrAg) and isolation on fungal media: Sabouraud's (SAB), brain-heart infusion agar (BHI) and canavanine-glycine-bromothymol blue (CGB) agar were analyzed. Unsatisfactory staining results by MM stain associated to negative titers by CrAg test, which FM stain confirmed that capsule-deficient Cryptococcus infections were observed in four cases. Eight isolated cases were identified as follows: six cases were infection with Cryptococcus neoformans and two cases were Cryptococcus gattii.

Unusual morphologies of Cryptococcus spp. in tissue specimens: report of 10 cases

Ten cases of cryptococcosis due to unusual microscopic forms of Cryptococcus sp. observed over a twenty-eight year period (1981-2009) are presented. The most important clinicopathological and laboratory data are tabulated. The uncommon forms of cryptococcal cells given are: structures resembling germ tube (one case), chains of budding yeasts (one case), pseudohyphae (two cases) and nonencapsulated yeast-like organisms (eight cases). The diagnosis was based on the histopathological findings. The causative organism was isolated and identified in seven cases; five were due to C. neoformans, and two to C. gattii. In addition, the importance of using staining histochemical techniques - Grocott's silver stain (GMS), Mayer's mucicarmine stain (MM) and Fontana-Masson stain (FM) - in the diagnosis of cryptococcosis is argued.

Whipple's disease: rare disorder and late diagnosis

Whipple's disease is a rare systemic infectious disorder caused by the bacterium Tropheryma whipplei. We report the case of a 61-year-old male patient who presented to emergency room complaining of asthenia, arthralgia, anorexia, articular complaints intermittent diarrhea, and a 10-kg weight loss in one year. Laboratory tests showed the following results: Hb = 7.5 g/dL, albumin = 2.5 mg/dL, weight = 50.3 kg (BMI 17.4 kg/m²). Upper gastrointestinal endoscopy revealed areas of focal enanthema in the duodenum. An endoscopic biopsy was suggestive of Whipple's disease. Diagnosis was confirmed based on a positive serum polymerase chain reaction. Treatment was initiated with intravenous ceftriaxone followed by oral trimethoprim-sulfamethoxazole. After one year of treatment, the patient was asymptomatic, with Hb = 13.5 g/dL, serum albumin = 5.3 mg/dL, and weight = 70 kg (BMI 24.2 kg/m²). Whipple's disease should be considered a differential diagnosis in patients with prolonged constitutional and/or gastrointestinal symptoms. Appropriate antibiotic treatment improves the quality of life of patients.

Técnicas histopatológicas no diagnóstico de criptococose por Cryptococcus deficiente de cápsula: relato de caso

Caso de criptococose por Cryptococcus deficiente de cápsula, no qual cultivo do espécime clínico e pesquisa do antígeno capsular no líquor e soro foram negativos. As técnicas histopatológicas foram: Hematoxilina-eosina, Grocott, Mucicarmim de Mayer e Fontana-Masson. Confirmou-se o diagnóstico do Cryptococcus deficiente de cápsula pela técnica de Fontana-Masson e pela imunofluorescência direta. É discutida a potencialidade das técnicas histoquímicas.

Bacteremias por Staphylococcus coagulase negativos oxacilina resistentes em um hospital escola na cidade de Santa Maria, Estado do Rio Grande do Sul

INTRODUÇÃO: Neste estudo, objetivou-se caracterizar a prevalência e o perfil de suscetibilidade de cepas de Staphylococcus coagulase negatives resistentes à oxacilina isoladas de culturas de sangue, em um hospital escola, localizado na Cidade de Santa Maria. Além disso, buscou-se comparar ao teste genotípico de referência, diferentes metodologias fenotípicas para a caracterização da resistência mediada pelo gene mecA. MÉTODOS: Após identificação (MicroScan® - Siemens), os isolados foram submetidos a testes de sensibilidade antimicrobiana a partir da difusão do disco e automação (MicroScan® - Siemens). A presença do gene mecA foi evidenciada através da técnica molecular de reação em cadeia da polimerase. RESULTADOS: A espécie prevalente foi Staphylococcus epidermidis (67%). O gene mecA foi detectado em 90% das cepas e conforme análise dos perfis de sensibilidade, observou-se um índice elevado de resistência a várias classes de antimicrobianos. Contudo, todos os isolados mostraram-se uniformemente sensíveis à vancomicina e tigeciclina. O disco de cefoxitina foi a metodologia fenotípica que melhor correlacionou-se com o padrão ouro. CONCLUSÕES: A análise da significância clínica de SCN isolados de hemoculturas e a detecção precisa da resistência à oxacilina representam fatores decisivos para a instituição correta da antibioticoterapia. Apesar da vancomicina constituir o tratamento usual na maioria dos hospitais brasileiros, tem a redução de seu emprego recomendada.

Evaluation of bacterial growth inhibition by mercaptopropionic acid in metallo-β-lactamase detection on multidrug-resistant Acinetobacter baumannii

INTRODUCTION: Metallo-β-lactamase (MBL) has been reported all over the world. METHODS: The inhibitory effect of mercaptopropionic acid (MPA) on bacterial growth was evaluated by comparison between disk diffusion and broth dilution methodology with determination of the minimum inhibitory concentration (MIC) for multidrug-resistant Acinetobacter baumanni strains. RESULTS: MPA significantly inhibited growth of the strains. CONCLUSIONS: The use of MPA can affect the results in phenotypic methods of MBL detection.

Detection of the viable myocardium. A perfusion scintigraphic study, before and after coronary bypass surgery in myocardial infarction patients

OBJECTIVE: To compare single-photon-emission computed tomography (SPECT) imaging scans using 201Tl and 99mTc-MIBI in detection of viable myocardium, in regions compromised by infarction. METHODS: Thirty-two (59.3±9.8 years old and 87% male) myocardial infarction patients were studied. All had Q waves on the ECG and left ventricle ejection fraction of <50%. They underwent coronary and left ventricle angiographies and SPECT before (including 201Tl reinjection) and after coronary artery bypass surgery (CABG). Improvement in perfusion observed after surgery was considered the gold standard for myocardial viability. RESULTS: Among 102 studied regions of the heart, there were 40 (39.2%) areas of transient perfusion defects in the conventional protocol with 201Tl and 52 (51.0%) after reinjection. Therefore, 12/62 (19.4%) more viable regions were identified by reinjection. Using 99mTc-MIBI, only 14 (13.7%) regions with transient defects were identified, all of which were seen also in 201Tl protocols. After surgery, 49 of a total of 93 regions analyzed (52.7%) were viable. Sensitivity, specificity, accuracy, positive and negative prediction values were, respectively, 201Tl SPECT scans - 65.3%, 90.9%, 77.4%, 88.9% and 70.2%, reinjection protocol with 201Tl scans - 81.6%, 81.8%, 81.7%, 83.3% and 80.0%; 99mTc-MIBI SPECT scans - 20.4%, 90.9%, 53.8%, 71.4% and 50.6%. Logistic regression demonstrated that the reinjection protocol with 201Tl was the best predictor of viability (P<0.001). CONCLUSION: Our data suggest the election of 201Tl for viability studies, especially when using the reinjection protocol.

Analysis of the effects of the continuous electrode paste band on precordial leads of the electrocardiogram

OBJECTIVE - To determine if the application of a continuous electrode paste band on precordial leads results in alteration of the electrocardiographic tracing as compared with an adequate amount of electrode paste, and if the former condition does not cause uniform morphologies from V1 to V6. METHODS - The amplitude and morphology of the electrocardiographic waves on the precordial leads in electrocardiographic tracings, which were performed with standard (control group) or excessive (continuous band) application of the electrode paste, were compared. RESULTS - None of the 106 patients studied showed uniformity of the QRS morphology from V1 to V6. The eletrocardiographic alterations identified in the tracings performed with a continuous electrode paste band that showed statistical significance in relation to the control group were the following: inversion of the P wave in V1; inversion of the T wave in V1, V2, and V3; appearance of R' waves in V1 and V2; disappearance of S waves in V1; appearance of S waves in V5 and V6; alterations in the amplitude of almost all waves, in all leads. CONCLUSION - Application of a continuous electrode paste band in the precordial leads may cause significant alterations in the electrocardiographic tracing obtained.

Electrocardiographic abnormalities in neurological diseases

OBJECTIVE: To evaluate electrocardiographic abnormalities in patients with neurologic diseases. METHODS: We studied 161 patients with neurologic disorders by analyzing the 12-lead electrocardiogram during the pathological process. An expert who did not know anything about the patients evaluated the traces. RESULTS: Neurological process included brain tumor (41%), stroke (27.3%), cerebral aneurysm (15.5%), subarachnoid hemorrhage (6.8%), subdural hemorrhage (5%), and head injury (4.4%). Electrocardiograms were normal in 61% of cases, and the most frequent abnormality was ventricular repolarization (23.7%). The presence of T waves (4.6%) and prolonged QT intervals (8.8%) was the most characteristic of brain injuries. CONCLUSION: We observed a lower incidence of electrocardiographic abnormalities than that described in the literature.

Co-administration of Apelin and T4 Protects Inotropic and Chronotropic Changes Occurring in Hypothyroid Rats

AbstractBackground:One of the most important thyroid hormone targets is the cardiovascular system. Hemodynamic changes, such as decreased resting heart rate (HR), myocardial contractility, and cardiac output, and increased diastolic pressure and systemic vascular resistance, have been observed in hypothyroid patients. Moreover, in these patients, ECG changes include sinus bradycardia and low voltage complexes (P waves or QRS complexes).Objective:This study aimed at evaluating the prophylactic effect of apelin on HR changes and QRS voltage that occur in propylthiouracil (PTU)-induced hypothyroid rats.Method:In this study, 48 adult male Wistar rats weighing 170-235g were randomly divided into 6 groups: Control group (normal saline ip injection + tap water gavage); P group (PTU 0.05%, in drinking water); A group (apelin 200 µg.kg-1.day-1, ip); PA group [co-administration of PTU and apelin]; PT group [co-administration of PTU + T4 (0.2 mg/g per day, gavage)]; and PAT group (co-administration of PTU, apelin and T4). All experiments were performed for 28 consecutive days, and then the animals were anesthetized with an ip injection of ketamine (80 mg/kg) and xylazine (12 mg/kg). Lead II electrocardiogram was recorded to calculate HR and QRS voltage.Results:Heart rate and QRS voltage increased more significantly in the hypothyroid group that consumed both apelin and T4 (201 ± 4 beat/min, 0.71 ± 0.02 mv vs. hypothyroid 145 ± 9 beat/min, 0.563 ± 0.015 mv; respectively).Conclusion:The co-administration of apelin and T4 showed a protective effect on QRS voltage and HR in PTU‑induced hypothyroid rats.

Contribuição ao estudo das Pasteurellas: pasteurella intermediária n. sp

Os autores decreveram uma nova bactéria - Pasteurella intermedia n. ap., obtida pela inoculação em cobaio de 2 cc. de sangue total de um indivíduo morto de bronco-pneumonia e suspeito de ter a forma grave de Tifo exantemático neotrópico. Têm a impressão que a Pasteurella marsupialis e a P. intermedia, constituem um grupo à parte, bem definido, dentro das Pasteurellas. Muito pequenas, de grande e persistente poder patogênico para os animais comuns de laboratório, mesmo quando as amostras das bactérias são conservadas pelos replantios em agar-comum, na temperatura e iluminação comum em laboratórios, durante anos. Estas duas Pasteurellas, ao contrário das demais, têm alto e inconfundível poder antígenico, para a formação de aglutininas e fixação do complemento e dão com constância uma "reação testicular" em cobaios machos, quando injetadas pela via intra-peritoneal, febre alta, esplenomegalia constante e às vezes notável, prestando-se à confusão para o diagnóstico diferencial e experimental com a raça VB do Tifo exantemático neotrófico no Brasil (Moléstia de Pisa, Gomes e Mayer).

Influência de elementos climáticos na mortalidade de cidades brasileiras

In this paper, preliminary to a series of investigations that the A. has the purpose to make about the influence of climatic factors particularly upon the prevalence of the most important acute infectious diseases in Brazil, he raises the question whether such factors do affect in this country the total death rates, as it is reasonable to suppose, according to what has been observed in temperate zones of northern and southern hemispheres. The inclusion of absolute humidity among other climatic factors to be dealt with seems justifiable according to Rogers and Stallybrass. Owing to scarcety of reliable data the A. was obliged to limit to a five-years period (1940-1944) the complete proposed investigation, which includes seven of the most important cities, scattered throughout the brazilian territory, from north to south - Belém, recife, Salvador, Rio, S. Paulo, Curitiba and Porto Alegre. Reference is made to their normal climatic conditions and monthly death-rates variations with their mean values and standard deviations. In a first part dealing with seasonal variations only for purposes of comparison, he points out that in there tropical cities of Brazil, without very clear seasonal differentiation, the curve of general mortality reached its highest point in austral autumn season and the remaining four (including Rio near the tropic) in the spring, with the exception of Curitiba, where the peak coincided with the summer season. He shows how such important causes of deaths, as diarrheas, common respiratory diseases and tuberculosis, whose seasonal distribution for each one of the seven cities is referred, may explain such seasonal variations. On a second part, a study is made of the general mortality distribution by four-months periods selected in accordance respectively with the highest or lowest values of rainfall and of mean temperature and humidity during period 1940-1944. Finally he compares the monthly waves of such climatic factors and the corresponding waves of total death - rates and finds through correlation coefficients 17 significant values with respect to their standard errors. Variations in the death - rates seemed to be perhaps more closely and uniformly associated with variations of mean humidity, as is indicated by coefficients ranging from + 0.3 to 0.6.

Doenças intestinais em cidades brasileiras: associação com elementos climáticos

Enteric fevers and dysenteries showed, during the period 1940-44, a summer prevalence in brazilian cities of the temperate zone. The distribu¬tion of the diseases by four-months periods, selected in acordance with the highest of lowest values of rainfall, mean temperature and absolute humidity induced to suppose that, in those cities, and also in tropical ones, both enteric fevers and dysenteries were closely associated with such climatic factors: enteric fevers mainly with absolute humidity and temperature, and dysenteries with humidity and rainfall. Correlation coefficients, statistically significant, have been obtained comparing monthly waves of climatic factors and corresponding waves of prevalence of the diseases. For enteric fevers, clear associations have been disclosed: with temperature variations in all temperate cities (coefficients ranging from + 0.42 to + 0.75, higher with mean temperature in the previous month) and in two of the four tropical cities (from + 0.26 to ± 0.30); with absolute humidity variations in cities of the first group (from + 0.51 to 0.71) and in the tropical city of Rio ( + 0.26 ± 0.12 and + 0.28 ± 0.12); and also with rainfall variations but only in two temperate cities (from + 0.28 to + 0.64). For dysenteric diseases, in cities of temperate zone similar associations have been found with absolute humidity (values of r, ranging from + 0.32 to + 0.45), with temperature (from + 0.26 to + 0.44); and with rainfall only in Curitiba ( + 0.25 ± 0.12). Recife (tropical city) yielded two significant values : r = — 0.27 ± 0.12 (correlation with mean temperature in the same month) and r = + 0.40 ± 0.11 and -h 0.37 ± 0.11 (between monthly morbidity rates and rainfall, respectively in the same month and in the previous one). Deaths by diarrhea and enteritis, in the cities of the temperate zone, prevailed in spring-summer seasons, also in four-months periods of highest temperature and humidity, for those cities and for the tropical ones, with the exception of Belem in which percentages were identical to those of opposite periods. Still with the exception of Belem, in all cities studied positive correla¬tion coefficients, statistically significant, have been obtained with temperature variations (ranging from + 0.25 to + 0.65 in tropical cities, and from + 0.47 to + 0.76 in temperate zone) and with humidity variations (from + 0.34 to + 0.44 in the first group, and from + 0.43 and + 0.74 in the second) . With rainfall, only Rio (in the tropical region) showed a significant value for r ( + 0.26 ±0:12); similarly in S. Paulo and Curitiba, the values ranged from + 0.46 to + 0.56, while in Porto Alegre there were found — 0.26 ± 0.12 and — 0.32 ± 012, for rainfall variations in the same and previous months.