Mini-percoll gradient may be used for frozen-thawed sperm selection in sheep / Gradiente de mini-Percoll pode ser utilizado na seleção espermática de sêmen congelado de ovinos

This study evaluated the effect of increasing centrifugal force and reducing centrifugation time and volume in Percoll protocols on ram sperm parameters. Commercial semen of Santa Inês rams were used and five treatments were performed: traditional Percoll and mini-Percoll (MP) techniques (I- 5000 x g, 5min; II- 2500 x g, 5min; III- 1250 x g, 5min; IV- 700 x g, 10min). At post-thawing (PT) and post-selection protocols (0h), samples were assessed for spermatozoa recovery rate, motility, plasma membrane (PM) integrity, sperm capacitation and morphology and incubated at 37 C for 1, 2 and 3h. The sperm recovery rate averaged 9.1±1.4%, and most motility parameters were similar (P> 0.05) among protocols. VCL (µm/s) was higher (P< 0.05) after MP-II, III and IV (66.1±4.5) than traditional Percoll (46.3±4.9). Capacitation status and PM integrity were similar (P> 0.05) among treatments. For the first time, we have demonstrated the reduction of the gradient volume and centrifugation time associated with an increase on centrifugation force at Percoll can be successfully used for frozen-thawed ram sperm selection. MP may be used instead of traditional Percoll, decreasing costs and semen handling time.(AU)

O presente estudo avaliou o efeito do aumento da força de centrifugação, bem como da redução do tempo de centrifugação e do volume do gradiente de Percoll em diferentes protocolos nos parâmetros espermáticos de ovinos. Foi utilizado sêmen comercial de carneiros da raça Santa Inês, e cinco tratamentos foram realizados: Percoll tradicional e quatro técnicas de mini-Percoll (I- 5000 x g, 5min; II- 2500 x g, 5min; III- 1250 x g, 5min; IV- 700 x g, 10min). Após o descongelamento e a seleção espermática em cada técnica utilizada (0h), amostras foram avaliadas quanto à taxa de recuperação espermática, motilidade, integridade de membrana plasmática, capacitação e morfologia. Ao final, foram incubadas a 37 ºC por uma, duas e três horas. A taxa de recuperação média (9,1±1,4%) e a maioria dos parâmetros de motilidade foram similares (P>0,05) entre os tratamentos. VCL foi maior (P<0,05) após MP-II, III e IV (66,1±4,5) quando comparados ao Percoll tradicional (46,3±4,9). O status da capacitação e a integridade de membrana foram similares (P>0,05) entre os tratamentos. Pela primeira vez, foi demonstrado que a redução do volume do gradiente utilizado e do tempo de centrifugação, associada com o aumento da força de centrifugação nos protocolos de Percoll, pode ser usada com sucesso na seleção espermática de sêmen congelado de ovinos. O mini-Percoll pode ser utilizado em alternativa à técnica de Percoll tradicional, diminuindo custos e tempo de manipulação do sêmen durante a técnica.(AU)

Major seminal plasma proteome of rabbits and associations with sperm quality.

The present study was conducted to describe the major seminal plasma proteome of rabbits and potential associations between seminal proteins and semen criteria. Semen samples were collected from 18 New Zealand adult rabbits, and seminal plasma proteins were analyzed by 2-D SDS-PAGE and tandem mass spectrometry. Sperm motility, vigor, concentration, morphology and membrane sperm viability were evaluated. Rabbits ejaculated 364 ±â€¯70 million sperm/ml, with 81 ±â€¯6.1% motile cells, 3.8 ±â€¯0.2 vigor and 66.7 ±â€¯2.5% sperm with normal morphology. Based on the viability and acrosome integrity assay, there were 65.8 ±â€¯2.5% live sperm with intact acrosome and most spermatozoa had both intact acrosome and functional membrane. On average, 2-D gels of rabbit seminal plasma had 232 ±â€¯69.5 spots, as determined by PDQuest software (Bio Rad, USA). Mass spectrometry allowed the identification of 137 different proteins. The most abundant proteins in rabbit seminal plasma were hemoglobin subunit zeta-like, annexins, lipocalin, FAM115 protein and albumin. The intensity of the spots associated with these five proteins represented 71.5% of the intensity of all spots detected in the master gel. Multiple regression models were estimated using sperm traits as dependent variables and seminal plasma proteins as independent ones. Also, sperm motility had positive association with beta-nerve growth factor and cysteine-rich secretory protein 1-like and a negative one with galectin-1. The percentage of rabbit sperm with intact membrane was related to seminal plasma protein FAM115 complex and tropomyosin. Then, the population of morphologically normal sperm in rabbit semen was positively linked to carcinoembryonic antigen-related cell adhesion molecule 6-like and down regulated by seminal plasma isocitrate dehydrogenase. Based on another regression model, the variation in the percentage of live sperm with intact acrosome was partially explained by the amount of leukocyte elastase inhibitor and the peptidyl-prolyl cis-trans isomerase A in the rabbit seminal fluid. The current study reports the identification of 137 proteins of rabbit seminal plasma. Major proteins of seminal secretion relate primarily to prevention of damages caused by lipid peroxide radicals and oxidative stress, membrane functionality, transport of lipids to the sperm membrane and temperature regulation. Moreover, finding seminal plasma proteins as indicators of semen parameters will improve assisted reproductive technologies.

Comparison of the Sensitivity of Three Methods for the Early Diagnosis of Sporotrichosis in Cats.

Sporotrichosis is caused by species of fungi within the Sporothrix schenckii complex that infect man and animals. In Rio de Janeiro, Brazil, an epidemic has been observed since 1998, with most of the cases being related to transmission from infected cats. Although the definitive diagnosis of feline sporotrichosis is made by fungal culture, cytopathological and histopathological examinations are used routinely, because the long culture period may delay treatment onset. However, alternative methods are desirable in cases of low fungal burden. Immunohistochemistry (IHC) has been described as a sensitive method for diagnosing human and canine sporotrichosis, but there are no reports of its application to cats. The aim of this study was to analyse the sensitivity of cytopathological examination (Quick Panoptic method), histopathology (Grocott silver stain) and anti-Sporothrix IHC by blinded comparisons, using fungal culture as the reference standard. Samples were collected from 184 cats with sporotrichosis that exhibited skin ulcers. The sensitivities of Grocott silver stain, cytopathological examination and IHC were 91.3%, 87.0% and 88.6%, respectively. Grocott silver stain showed the best performance. IHC showed high sensitivity, as did cytopathological examination and these may be considered as alternative methodologies. When the three methods were combined, the diagnosis was established in 180 (97.8%) out of 184 cases. Taken together, these findings indicate the need to implement these methods as routine tools for the early diagnosis of sporotrichosis in cats, notably when fungal culture is not available.

Proteomic analysis of seminal plasma from locally-adapted "Curraleiro Pé-Duro bulls" (Bos taurus): identifying biomarkers involved in sperm physiology in endangered animals for conservation of biodiversity.

The present study was aimed at evaluating the seminal plasma proteins and sperm parameters of Curraleiro Pé-Duro bulls. Semen was collected from 10 bulls by electroejaculation, and sperm parameters were evaluated in fresh and frozen-thawed semen. Seminal plasma proteins were analyzed by 2-D SDS-PAGE and mass spectrophotometry. Tools in computational biology were used to generate bioinformatic knowledge and evaluate gene ontology, protein-protein interactions, phylogenetic trees and multiple sequence alignments. Sperm motility in fresh and frozen-thawed semen was 78.8±1.8% and 21.2±1.6%, respectively. Pearson's correlations were evaluated (p<0.05). Sperm motility and vigor in fresh semen were correlated with clusterin, TIMP2 and cathepsin S (r=0.64-0.71) and sperm defects were related to inhibitor of carbonic anhydrase and BSP 5 (r=0.78-0.80). Clusterin, BSP 5, alpha-enolase, creatine kinase M-type, glyceraldehyde-3-phosphate dehydrogenase, BSP 3, albumin, and 5'-nucleotidase and legumain were correlated with acrosome intact live sperm (r=0.80-0.64). Associations were detected between sperm vigor and spermadhesin 1 (r=-0.89), and between sperm defects in fresh semen and spermadhesin 1 and clusterin (r=-0.81). Sperm motility in frozen-thawed semen was associated with BSP 1, spermadhesin 1, clusterin and spermadhesin Z13 (r=0.64-0.85). The percent of motile sperm after freeze-thawing was negatively correlated (r=-0.64) with the amount of spermadhesin 1 in the seminal plasma. Based on in silico analysis, TIMP2 interacted with BSP1, BSP3, BSP5 and metalloproteinases. Molecular functions of proteins associated with sperm parameters were binding, catalytic activity and enzymatic regulation. Amino acid sequences of spermadhesin 1 and BSP 1 from Bos taurus, and other domestic species were similar. Phylogenetic tree analysis demonstrated that clusterin from Bos taurus was related to Ovis aries and domains of clusterin, spermadhesin 1, BSP 1 and inhibitor of carbonic anhydrase were conserved as well. In summary, specific seminal proteins are associated with sperm parameters of locally-adapted bulls. Use of the endangered mammalian as a model may assist in understanding aspects of evolutionary adaptations and could improve assisted reproductive biotechnologies.

Sensitivity and specificity of serological tests, histopathology and immunohistochemistry for detection of Toxoplasma gondii infection in domestic chickens.

Since free-range chickens are important for the epidemiology of toxoplasmosis, this study evaluated the sensitivity and specificity of different laboratory techniques for the diagnosis of Toxoplasma gondii in these animals. Serum samples from 135 adult domestic chickens were tested for anti-T. gondii antibodies by the indirect fluorescent antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA), modified agglutination test (MAT), and indirect hemagglutination test (IHAT). Tissue samples from all animals were analyzed by histopathology, immunohistochemistry and mouse bioassay (gold standard). Fifty-four chickens were positive for T. gondii in the bioassay. The sensitivity and specificity of the different tests were, respectively, 85% and 56% for ELISA; 80% and 52% for IFAT; 76% and 68% for MAT; 61% and 80% for IHAT; 7% and 98% for immunohistochemistry, and 6% and 98% for histopathology. The MAT was the most effective method for the diagnosis of T. gondii infection in chickens, followed by ELISA. Histopathology and immunohistochemistry are useful tools for the diagnosis of T. gondii infection in chickens due to their specificity.

Parameters of the reproductive tract, spermatogenesis, daily sperm production and major seminal plasma proteins of tropically adapted morada nova rams.

This study describes the reproductive parameters of Morada Nova rams, a breed of hair sheep from Brazil and with unique adaption to tropical environments. At 42 weeks of age, 15 rams were subjected to semen collection and, 1 week later, animals were slaughtered for collection of testes, epididymis and accessory sex glands. We conducted 2-D electrophoresis of seminal plasma proteins and major spots of stained gels were identified by LC-MS/MS. Total RNA was isolated from testis, epididymis and vesicular glands and subjected to qPCR. At slaughter, scrotal circumference and testicular weight were 27.5 ± 0.5 cm and 109.5 ± 6.0 g, respectively. Seminiferous tubule (ST) diameter was 188.3 ± 4.0 µm and each testis contained 1.9 ± 0.1 Sertoli cells (×10(9) ). Each Sertoli cell supported 0.1 ± 0.01 A spermatogonia, 3.0 ± 0.2 pachytene spermatocytes and 7.7 ± 0.5 round spermatids/tubule cross section. Daily sperm production reached 5.6 × 10(6)  cells/g of testis parenchyma. Testis size appeared as indicative of ST diameter and associated with epididymal measurements, as well as with the population of round spermatids and Sertoli cells/testis. Rams with heavier testes had greater daily sperm production and more Sertoli cells/testis. We detected 90.9 ± 9.6 spots per 2-D gel of seminal plasma. Major seminal proteins were identified as ram seminal vesicle proteins at 14 and 22 kDa, representing 16.2% and 12.8% of the total intensity of valid spots in the gels, respectively. Expression of both genes was greater in the vesicular glands as compared to testis and epididymis. Pixel intensity for those proteins in the 2-D gels was significantly correlated with seminal vesicle weight. This is the first description of the basic reproductive aspects of Morada Nova rams, including protein profiles of their seminal plasma. These findings will allow a better understanding of their reproductive physiology.

Desempenho produtivo e econômico do confinamento de bovinos zebuínos alimentados com três dietas de alto concentrado / Economic performance of zebu cattle under feedlot fed with three high concentrate diets

Estudaram-se o desempenho produtivo e a viabilidade econômica no confinamento de 24 bovinos, inteiros, com idade média de 23 meses, sendo 12 Nelore (NEL) e 12 F1 Nelore x Brahman (NBR), divididos em três dietas: SIL - silagem de milho e ração concentrada; PEL - exclusiva de pellets; GRN - milho grão inteiro e pellets. O experimento foi realizado em um delineamento inteiramente ao acaso, em esquema fatorial 2x3, com quatro repetições. Os animais NBR apresentaram resultados semelhantes aos da raça NEL quanto aos desempenhos alimentar e econômico, para todas as variáveis analisadas durante o confinamento. Entre as três dietas, a dieta PEL, apesar de não ser a de maior custo em R$/kg, obteve o menor desempenho, com maior COT/kg, menor RT e menores margens líquidas. A dieta SIL apresentou os menores COT/kg e custo/kg de carcaça, com maiores receitas e margens líquidas. A dieta GRN obteve resultados econômicos semelhantes à SIL, porém obteve menor GMD em relação à SIL, além de ter o custo de dieta (R$/kg) mais oneroso em relação aos três tratamentos. Todas as dietas foram economicamente viáveis ao final do experimento, com ML/Kg de carcaça médios de 2,12, 0,35 e 1,09 para SIL, PEL e GRN, respectivamente, no entanto a dieta SIL foi a de melhor desempenho produtivo e econômico.

The production performance and economic viability of a feedlot of 24 cattle, with an average age of 23 months, 12 Nellore (NEL) and 12 F1 Nellore x Brahman (NBR), divided into three diets: SIL - corn silage and concentrate diet; PEL - exclusive of pellets; GRN - corn and whole grain pellets were studied. The experiment was conducted in a completely randomized scheme in a 2x3 factorial with four replications. The NBR animals showed similar results to the NEL in performance and economic viability for all variables analyzed. Among the diets, the PEL diet, although not the highest diet cost in R$/kg, had the lowest performance with higher TOC/kg, lower TR and lower net margins. The SIL diet had the lowest TOC/kg and cost/kg of carcass, with higher revenues and net margins. The economic results obtained for the GRN diet were similar to SIL, but had lower ADG compared to SIL and highest diet cost (R$/kg) among all. The diets were economically viable at the end of the trial, with average net margin/kg carcass of 2.12, 0.35 and 1.09 for SIL, FEP and GRN, respectively; however the SIL diet presented better productive and economical performance.

Influence of the nutritional status in the clinical and therapeutical evolution in adults and elderly with American Tegumentary Leishmaniasis.

The objective of this study is to describe the nutritional status of adult and elderly patients with American Tegumentary Leishmaniasis (ATL). It was conducted a longitudinal study in 68 adult and elderly patients with ATL treating at the Surveillance Leishmaniasis Laboratory at the Evandro Chagas Clinical Research Institute, Oswaldo Cruz Foundation (Fiocruz), from 2009 to 2012. The nutritional assessment included the body mass index (BMI) and serum albumin levels. The clinical evolution (epithelialization and wound healing) was measured up to two years after ATL treatment. Most of the sample was composed of men (71%), adults (73%), with household income of 1-5 minimum wages (79%), and incomplete elementary school (48.5%). The predominant ATL form was cutaneous (72%), and 39% presented comorbidities, the most frequent was hypertension (30.8%). The most prevalent clinical and nutritional events were: recent decrease in food intake (23.9%); nasal obstruction (22.1%); oral ulcer (14.7%), anorexia and dysphagia (13.2% each) and odynophagia (10.3%). The total healing time was 115.00 (IR=80-230) days for skin lesions, and 120.00 (IR=104.50-223.50) days for mucous membrane lesions. Low body weight in 10%, and hypoalbuminemia in 12% of the patients have been observed. Low body weight was associated with age, mucosal leishmaniasis (ML), nasal obstruction, recent decrease in food intake and hypoalbuminemia. As for serum albumin depletion, association with the ML, dyspnea, dysphagia, odynophagia, recent decrease in food intake, absence of complete healing of the skin lesions, and increased healing time for mucous membrane lesions, was observed. The ML and their events that affect the alimentary intake have been related to the impairment of the nutritional status. Additionally, serum albumin depletion negatively affected the healing of the lesions, suggesting that a nutritional intervention can increase the effectiveness of the ATL treatment.

Falls in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP).

STUDY DESIGN: Cross-sectional study. OBJECTIVES: To determine the prevalence of falls in human T-cell lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients and possible factors associated to their occurrence. SETTING: Instituto de Pesquisa Clínica Evandro Chagas, Fundação Oswaldo Cruz (FIOCRUZ) - Brazil. METHODS: Thirty-six HAM/TSP patients able to walk at least 20 m were assessed by a questionnaire. Data regarding gender, age, duration of disease (DD), HTLV-I proviral load (HPL), frequency of physical activity (FCA), use of walking aids, functional ambulation level, the number of falls and associated injuries in the last year were reviewed. Multiple correspondence analysis was used to group characteristics of this sample according to the fall occurrence. RESULTS: The prevalence of falls was 63.9% and we observed injuries in 47.8% of the cases. Four groups were identified in the descriptive analysis. One group was formed by faller individuals, men <60 years, independent ambulation, FCA≥3 times per week and HPL <6.6 copies per 100 cells (group B). The other one comprised non-faller patients, women ≥60 years, restricted ambulation, DD ≥7 years, use of orthosis, FCA 0-1 time per week and HPL ≥6.6 copies per 100 cells (group D). The others two groups comprised individuals that did not use orthosis (group A) and those that FCA was two times per week and DD <7 years (group C). CONCLUSION: Falls occur in roughly two-thirds of ambulatory HAM/TSP patients and are associated with significant morbidity. Further studies with a larger number of patients are necessarily to identify risk factors in order to elaborate specific programs to prevent falls in this population.

Evaluation of serological cross-reactivity between canine visceral leishmaniasis and natural infection by Trypanosoma caninum.

In order to evaluate if the presence of Trypanosoma caninum can lead to a confuse diagnosis of canine visceral leishmaniasis (CVL), we investigated the serological status of dogs infected by T. caninum and assessed the serological cross-reactivity with CVL. A set of 117 serum samples from dogs infected by T. caninum, Leishmania chagasi and not infected dogs (n=39 in each group) was tested using commercial kits--indirect immunofluorescence (IFI-LVC), ELISA (EIE-LVC) and immunochromatographic test (DPP)--and in house tests with T. caninum (IIF-Tc and ELISA-Tc) and L. chagasi antigens (IIF-Lc and ELISA-Lc). IIF-Tc and ELISA-Tc presented sensitivity of 64.1% and 94.9% and specificity of 23.1% and 35.9%, respectively. The sensitivity of the IFI-LVC, EIE-LVC and DPP tests was 100% and the specificity was 70.5%, 68% and 97.5% respectively. The concordance between the tests was considered as satisfactory. The specificities of IFI-LVC, EIE-LVC and DPP were higher when the group Tc was excluded, with significant values for IFI-LVC (χ2=4.36, P-value=0.036), thus suggesting that the infection by T. caninum can confuse the diagnosis of CVL.

Avaliação de espermatozoides caprinos congelados em meio à base de água de coco em pó (ACP-101®) ou TRIS / Evaluation of goat spermatozoa frozen in media based on powder coconut water media based (ACP-101®) or TRIS

Compararam-se as características cinéticas e morfológicas de espermatozoides caprinos congelados nos meios à base de ACP-101® e TRIS. Os diluentes utilizados foram: ACP-101® (+ 2,5 por cento gema ovo + 7 por cento glicerol) e TRIS (+ 20 por cento gema ovo + 6,8 por cento glicerol). Quarenta e oito ejaculados de quatro bodes foram coletados, avaliados, divididos em duas alíquotas e diluídos nos meios ACP-101® e TRIS, respectivamente, posteriormente congelados e, após 30 dias, descongelados. A avaliação da motilidade espermática por computador foi realizada aos 5, 60 e 120 minutos pós-descongelação. As características de motilidade espermática analisadas foram: motilidade total (MT) ( por cento) e progressiva (MP) ( por cento), velocidades média do trajeto do espermatozoide (VAP) (µm/s) e linear (VSL) (µm/s) e população de espermatozoides rápidos (ER) ( por cento). As avaliações de morfologia espermática quantificaram a porcentagem de espermatozoides normais (N) e as alterações da cabeça (AC), da peça intermediária (API) e do flagelo (AF), aos cinco e 120 minutos pós-descongelação. O diluente TRIS apresentou resultados cinéticos mais elevados que o ACP-101® aos 60 e 120 minutos pós-descongelação. As AC aos 120 minutos pós-descongelação foram mais altas nos espermatozoides congelados em ACP-101®. Conclui-se que o diluente TRIS promoveu maior viabilidade in vitro dos espermatozoides caprinos pós-descongelação.

The aim of the work was to compare kinetic and morphologic characteristics of goat sperm frozen in diluent media based on ACP-101® and TRIS. The employed diluents were: ACP-101® (+ 2.5 percent egg yolk + 7 percent glycerol) and TRIS (+ 20 percent egg yolk + 6.8 percent glycerol). Forty eight ejaculates from four bucks were collected, assessed and divided into two aliquots and diluted into the experimental treatments ACP-101® and TRIS. The samples were frozen and after 30 days, thawed. The computer assisted spermatic motility evaluations were placed into 5, 60 and 120 minutes post-thawing. The motion parameters assessed were: total motility (MT) ( percent), progressive motility (MP) ( percent), average path velocity (VAP) (µm/s), straight linear velocity (VSL) (µm/s), and population of rapid spermatozoa (ER) ( percent). The morphologic parameters: normal spermatozoa (N), head alteration (AC), intermediary piece alteration (API), tail alteration (AF) were evaluated at 5 and 120 min post-thawing. The media based on TRIS showed kinetic results significantly superior to ACP-101® at 60 and 120min. After 120min post-thawing the AC was higher in frozen sperm in media based on ACP-101®. The TRIS media promoted better goat spermatozoa in vitro viability post-thawing.

An MD-PhD program in Brazil: students’ concepts of science and of common sense

In 1995, a pioneering MD-PhD program was initiated in Brazil for the training of medical scientists in experimental sciences at the Federal University of Rio de Janeiro. The program’s aim was achieved with respect to publication of theses in the form of papers with international visibility and also in terms of fostering the scientific careers of the graduates. The expansion of this type of program is one of the strategies for improving the preparation of biomedical researchers in Brazil. A noteworthy absence of interest in carrying out clinical research limits the ability of young Brazilian physicians to solve biomedical problems. To understand the students’ views of science, we used qualitative and quantitative triangulation methods, as well as participant observation to evaluate the students’ concepts of science and common sense. Subjective aspects were clearly less evident in their concepts of science. There was a strong concern about "methodology", "truth" and "usefulness". "Intuition", "creativity" and "curiosity" were the least mentioned thematic categories. Students recognized the value of intuition when it appeared as an explicit option but they did not refer to it spontaneously. Common sense was associated with "consensus", "opinion" and ideas that "require scientific validation". Such observations indicate that MD-PhD students share with their senior academic colleagues the same reluctance to consider common sense as a valid adjunct for the solution of scientific problems. Overcoming this difficulty may be an important step toward stimulating the interest of physicians in pursuing experimental research.

An MD-PhD program in Brazil: students' concepts of science and of common sense.

In 1995, a pioneering MD-PhD program was initiated in Brazil for the training of medical scientists in experimental sciences at the Federal University of Rio de Janeiro. The program's aim was achieved with respect to publication of theses in the form of papers with international visibility and also in terms of fostering the scientific careers of the graduates. The expansion of this type of program is one of the strategies for improving the preparation of biomedical researchers in Brazil. A noteworthy absence of interest in carrying out clinical research limits the ability of young Brazilian physicians to solve biomedical problems. To understand the students' views of science, we used qualitative and quantitative triangulation methods, as well as participant observation to evaluate the students' concepts of science and common sense. Subjective aspects were clearly less evident in their concepts of science. There was a strong concern about "methodology", "truth" and "usefulness". "Intuition", "creativity" and "curiosity" were the least mentioned thematic categories. Students recognized the value of intuition when it appeared as an explicit option but they did not refer to it spontaneously. Common sense was associated with "consensus", "opinion" and ideas that "require scientific validation". Such observations indicate that MD-PhD students share with their senior academic colleagues the same reluctance to consider common sense as a valid adjunct for the solution of scientific problems. Overcoming this difficulty may be an important step toward stimulating the interest of physicians in pursuing experimental research.

Interobserver agreement on signs and symptoms of patients with acute febrile illness.

PURPOSE: To assess the interobserver agreement on clinical history and physical examination when using a semi-structured questionnaire to evaluate patients with an acute febrile illness (AFI). METHODS: A cross-sectional study was conducted with outpatients aged 12 years and over, presenting with an AFI defined as fever up to 7 days and no evident focus of infection. Clinical data were collected independently by two physicians using a semi-structured questionnaire. Interobserver agreement was estimated using kappa coefficients with a 95% confidence interval (CI). RESULTS: A total of 140 patients (age range 13-73 years; 56.4% females) were enrolled. All symptoms showed weighted kappa values significantly greater than 0.6, indicating an at least substantial agreement. As most physical signs were infrequent and of mild intensity, they were recoded and analyzed as absent/present. Of the signs with prevalence ≥15%, exanthema, pallor, lymph node enlargement, and eye congestion showed agreements significantly greater than 0.6, while kappa confidence limits for pharyngeal erythema and dehydration included values classified as regular. CONCLUSIONS: High agreement was observed for most of the clinical data assessed, and symptom grading was feasible. Some physical findings were rare and their inclusion in a structured form may not be justified in this setting. The questionnaire application showed good reliability for the most frequent signs and symptoms and may prove to be useful at gathering data for surveillance and research at sentinel sites.

Risk factors associated with dizziness during treatment of mucosal leishmaniasis with meglumine antimoniate: 16-year retrospective study of cases from Rio de Janeiro, Brazil.

OBJECTIVE: To evaluate dizziness in patients receiving meglumine antimoniate for the treatment of mucosal leishmaniasis. MATERIALS AND METHODS: We retrospectively studied 127 patients treated at the Laboratory of Leishmaniasis Surveillance, Evandro Chagas Clinical Research Institute, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil, between 1 January 1989 and 31 December 2004. RESULTS: A low dose of meglumine antimoniate (5 mg/kg/day) was used in 86.6 per cent of patients; a dose of 10 mg/kg/day or higher was used in 13.4 per cent of patients. Dizziness was reported by 4.7 per cent of patients. The adjusted odds ratios were 7.37 for dizziness in female patients, 4.9 for dizziness in patients aged 60 years or older, and 7.77 for dizziness in the presence of elevated serum lipase. CONCLUSION: We suggest that dizziness may be a side effect of meglumine antimoniate, particularly in elderly individuals, in females and in patients with elevated serum lipase.

Comparative histopathological study of sporotrichosis and American tegumentary leishmaniasis in dogs from Rio de Janeiro.

Pyogranulomatous lesions from 80 dogs with sporotrichosis and 26 dogs with American tegumentary leishmaniasis (ATL) were compared microscopically in order to identify features that would support the diagnostic suspicion and direct the subsequent search for the aetiological agent of either infection. Odds ratios and their respective 95% confidence intervals were calculated in order to evaluate the impact of the microscopical findings on the diagnosis of either disease. Lesions with well-formed granulomata were 14 times more likely to be due to sporotrichosis than ATL. Marked neutrophil infiltration into granulomata was 12.26 times more likely to be associated with sporotrichosis when compared with lesions having mild neutrophilic infiltration. Absence of lymphocytes and macrophages in the peripheral infiltrate was associated with a 9.71 and 4.93 higher chance, respectively, of being sporotrichosis rather than ATL compared with lesions where these cells were present. Lesions with a perivascular, perifollicular and interstitial peripheral inflammatory infiltrate were 5.48 times more likely to be due to sporotrichosis than ATL when compared with lesions with a diffuse peripheral infiltrate. Histopathological analysis may therefore contribute to the diagnosis of sporotrichosis or ATL skin lesions in dogs since this method permits the identification of features that direct the diagnostic suspicion, thus facilitating the search for the aetiological agent in histological sections, permitting the precise request of subsequent tests and thereby reducing costs and time taken to achieve a definitive diagnosis and the initiation of appropriate therapy.

Histopathology of canine sporotrichosis: a morphological study of 86 cases from Rio de Janeiro (2001-2007).

The present study reports the histopathological findings of 86 skin lesions of dogs with sporotrichosis from Rio de Janeiro. Suppurative granulomatous inflammation was the predominant finding and was observed in 76 (88.37%) cases. Plasma cells surrounding the suppurative granulomas were detected in 68 (89.5%) cases and an inflammatory infiltrate at the periphery of these granulomatous lesions was observed in 63 (82.9%). Fungus-specific staining revealed yeast cells compatible with Sporothrix schenckii in 36 cases. These fungal elements were only detected in lesions characterized by suppurative granulomatous inflammation. Thus, specific staining of serial sections is recommended in the case of dogs with skin lesions whose histopathological presentation is consistent with sporotrichosis. However, due to the generally small number of yeast cells in lesions, the hypothesis of sporotrichosis should not be ruled out even if the result is negative, especially in epidemic areas where correlation with epidemiological data is particularly useful.