Abstract Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at -80oC for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at -20oC for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at -80oC for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at -20oC.

Abstract Background: Cryptoccocal meningitis continues to present high incidence among AIDS patients. The treatment of choice is the synergistic combination of flucytosine (5-FC) with amphotericin B deoxycholate (AmBd) or its lipid formulations. However, 5-FC is unavailable in many countries and AmB demands hospitalization. The combination of AmB with the fungistatic fluconazole (FLC) or the use of high FLC daily doses alone became the choice. Nonetheless, sterilization of cerebrospinal fluid is delayed with FLC monotherapy, mainly with high fungal burden. These findings suggest the search for new antifungal compounds, such as liriodenine. Methods: Liriodenine antifungal activity was evaluated by three procedures: determining the minimum inhibitory concentration (MIC) on 30 strains of the Cryptococcus neoformans (C. neoformans) complex and 30 of the Cryptococcus gattii (C. gattii) complex, using EUCAST methodology and amphotericin B deoxycholate as control; performing the time-kill methodology in two strains of the C. neoformans complex and one of the C. gattii complex; and injury to cryptococcal cells, evaluated by transmission electron microscopy (TEM). Liriodenine absorption and safety at 0.75 and 1.50 mg.kg-1 doses were evaluated in BALB/c mice. Results: Liriodenine MICs ranged from 3.9 to 62.5 μg.mL-1 for both species complexes, with no differences between them. Time-kill methodology confirmed its concentration-dependent fungicidal effect, killing all the strains below the limit of detection (33 CFU.mL-1) at the highest liriodenine concentration (32-fold MIC), with predominant activity during the first 48 hours. Liriodenine induced severe Cryptococcus alterations - cytoplasm with intense rarefaction and/or degradation, injury of organelles, and presence of vacuoles. Liriodenine was better absorbed at lower doses, with no histopathological alterations on the digestive tract. Conclusion: The fungicidal activity confirmed by time-kill methodology, the intense Cryptococcus injury observed by TEM, the absorption after gavage administration, and the safety at the tested doses indicate that the liriodenine molecule is a promising drug lead for development of anticryptococcal agents.

Resumo Introdução: A adenotonsilectomia é um procedimento que, por ser invasivo, pode incentivar a busca de informações pela população leiga por meio de ferramentas de busca online. A qualidade dessas informações, no entanto, requer uma avaliação devido à grande diversidade do conteúdo disponível. Objetivo: Avaliar a qualidade dos websites de maior acesso voltados à orientação de pais/responsáveis de crianças às quais a adenotonsilectomia é indicada, em termos de princípios éticos, legibilidade e abrangência. Método: Os websites contidos nas primeiras 10 páginas obtidas pela busca das palavras-chave “retirada”, “amígdalas” e “adenoides” foram selecionados sob critérios de inclusão e de exclusão e avaliados com as ferramentas Teste de Facilidade de Leitura Flesch e Health on the Net Code (HON Code). Também foi feita avaliação de conteúdo por dois avaliadores independentes. Os dados foram descritos, a concordância interavaliador foi calculada pelo coeficiente Kappa. Resultados: Foram encontrados 34 websites, dos 100 analisados, que se adequavam aos critérios de inclusão e exclusão com as ferramentas Google e Yahoo! Pelo Índice de Facilidade de Leitura Flesch 16 (47%) páginas foram consideradas de leitura razoavelmente difícil/difícil. A maioria atendeu a menos da metade dos quesitos éticos analisados e a média de abrangência geral foi considerada como insuficiente. Conclusão: Demonstrou-se uma realidade deficitária em termos de qualidade geral (abrangência, legibilidade e princípios éticos) associada aos websites disponíveis para pais ou cuidadores sobre o procedimento da cirurgia das tonsilas em crianças.

Abstract Introduction: Adenotonsillectomy is an invasive surgical procedure, which may encourage the search for information by the lay population through online search engines. The quality of this information, however, requires an evaluation due to the wide diversity of the available content. Objective: To evaluate the quality, in terms of ethical principles, readability and comprehensiveness, of the most accessed websites concerning guidance to parents/guardians related to their children’s recommended adenotonsillectomy. Methods: The websites mentioned on the first 10 pages obtained after the search using “removal”, “tonsils” and “adenoids” as keywords, after applying the inclusion and exclusion criteria, were selected. All were assessed using the Flesch Readability Index and Health on the Net Code tools, in addition to an evaluation of the content by two independent evaluators. The data were described, and the inter-rater agreement was calculated by the Kappa coefficient. Results: 34 websites were found, of the 100 assessed ones, which met the inclusion and exclusion criteria using the Google and Yahoo! Tools. Sixteen (47%) pages were considered reasonably difficult/difficult to read according to the Flesch Readability Index. Most of them met less than half of the analyzed ethical requirements according to the Health on the Net Code, and the overall comprehensiveness average was considered insufficient. Conclusion: A deficit of overall quality (comprehensiveness, readability, and ethical principles) was demonstrated for the websites available to parents or guardians about the adenotonsillectomy procedure in children.

Abstract Background: The present study was carried out aiming to evaluate the impact of laryngeal sequelae on the quality of life of treated paracoccidioidomycosis (PCM) patients. Methods: This cross-sectional study was conducted at the Otorhinolaryngology Outpatient Clinic of the University Hospital, Federal University of Mato Grosso do Sul, Brazil. Thirty-two PCM patients considered clinically and immunologically cured were included: 16 with laryngeal involvement during the active phase of the disease (laryngeal PCM group) and 16 without laryngeal involvement (control group). They were submitted to structured interview, otorhinolaryngology examination, videolaryngoscopy, videoendoscopic swallowing study, completed two questionnaires for voice self-assessment - Voice-related Quality of Life (V-RQOL) and Voice Handicap Index (VHI) - and were asked to score their voices on a scale from zero to 10 (self-assessment of vocal quality). Results: Dysphonia was present in 50% of the cases. Patients with laryngeal PCM presented worse voice-related quality of life scores on the V-RQOL and poorer vocal quality self-assessment than the control group. No significant differences in the VHI were found between the groups. None of the participants developed dysphagic sequelae, although some minor changes were observed on videoendoscopic examination. Conclusion: There were no dysphagia complaints and only a few mild changes were found on the fiberoptic endoscopic evaluation of swallowing, suggesting that this evaluation should be performed only in specific cases. Patients with laryngeal involvement presented worse V-RQOL and self-assessment voice quality. This study contributes to the current knowledge of the functional assessment of the larynx affected by PCM and the impact of dysphonia on quality of life.

BACKGROUND The elimination of malaria depends on the blocking of transmission and of an effective treatment. In Brazil, artemisinin therapy was introduced in 1991, and here we present a performance overview during implementation outset years. METHODS It is a retrospective cohort (1991 to 2002) of patients treated in a tertiary centre of Manaus, with positive microscopic diagnosis of Plasmodium falciparum malaria, under treatment with using injectable or rectal artemisinin derivatives, and followed over 35-days to evaluate parasite clearance, death and recurrence. FINDINGS This cohort outcome resulted 97.6% (1554/1593) of patients who completed the 35-day follow-up, 0.6% (10/1593) of death and 1.8% (29/1593) of follow-up loss. All patients that died and those that presented parasitaemia recurrence had pure P. falciparum infections and received monotherapy. Considering patients who completed 35-day treatment, 98.2% (1527/1554) presented asexual parasitaemia clearance until D4 and 1.8% (27/1554) between D5-D10. It is important to highlight that had no correlation between the five treatment schemes and the sexual parasite clearance. Finally, it is noteworthy that we were able to observe also gametocytes carriage during all follow-up (D0-D35). MAIN CONCLUSIONS Artemisinin derivatives remained effective in the treatment of falciparum malaria during first 12-years of use in north area of Brazil.

Resumo Justificativa: O jejum pré-operatório adequado é fundamental para prevenir a aspiração pulmonar do conteúdo gástrico. Nossa proposta foi avaliar a dinâmica ultrassonográfica do conteúdo gástrico após a ingestão de alimentos líquidos ou sólidos em voluntários sadios e confrontá-la com as diretrizes atuais para os períodos de jejum no pré-operatório. Métodos: Um estudo prospectivo, cruzado e avaliador-cego foi feito com 17 voluntários saudáveis de ambos os sexos. Cada participante jejuou por 10 horas e foi submetido a uma ultrassonografia gástrica na fase basal, ingestão de 400 mL de água de coco ou 355 g de sanduíche de carne e avaliações gástricas ultrassonográficas foram feitas após 10 minutos e a cada hora até o estômago estar completamente vazio. Resultados: Na fase basal, todos os participantes estavam com o estômago vazio. Aos 10 minutos, o conteúdo gástrico [média + desvio-padrão (DP)] foi de 240,4 + 69,3 e 248,2 + 119,2 mL para alimentos líquidos e sólidos, respectivamente (p > 0,05). Os tempos médios de esvaziamento gástrico + DP foram de 2,5 + 0,7 e 4,5 + 0,9 horas para alimentos líquidos e sólidos, respectivamente (p < 0,001). Para a bebida, o estômago ficou completamente vazio em 59% e 100% dos sujeitos após duas e quatro horas; para o sanduíche, o estômago ficou completamente vazio em 65% e 100% dos sujeitos após quatro e sete horas, respectivamente. Conclusões: A dinâmica ultrassonográfica do volume gástrico para água de coco e sanduíche de carne resultou em tempos totais de esvaziamento gástrico maiores e menores, respectivamente, do que os sugeridos pelas diretrizes atuais para o jejum pré-operatório.

Abstract Background: Adequate preoperative fasting is critical in preventing pulmonary aspiration of gastric content. We proposed to study the sonographic gastric content dynamics after the ingestion of liquid or solid food in healthy volunteers and confront it with current guidelines for preoperative fasting times. Methods: We performed a prospective, crossover, evaluator-blinded study involving 17 healthy volunteers of both sexes. Each participant fasted for 10 h and was subjected to a baseline gastric ultrasound, intake of 400 mL of coconut water or a 145 g, 355 kcal meat sandwich, and sonographic gastric evaluations after 10 min and every hour until the stomach was completely empty. Results: At baseline, all subjects had an empty stomach. At 10 min, gastric content [mean + standard deviation (SD)] was 240.4 + 69.3 and 248.2 + 119.2 mL for liquid and solid foods, respectively (p > 0.05). Mean + SD gastric emptying times were 2.5 + 0.7 and 4.5 + 0.9 h for liquid and solid foods, respectively (p < 0.001). For the drink, the stomach was completely empty in 59% and 100% of the subjects after two and four hours, and for the sandwich, 65% and 100% of the subjects after four and seven hours, respectively. Conclusions: Sonographic gastric dynamics for coconut water and a meat sandwich resulted in complete gastric emptying times higher and lower, respectively, than those suggested by current guidelines for preoperative fasting.

BACKGROUND The main clinical forms of paracoccidioidomycosis (PCM) are the acute/subacute form (AF) and the chronic form (CF), and they both display considerable clinical variability. The immune responses of PCM patients, during and after treatment, remain neglected, mainly in the case of CF patients, due to the high prevalence of pulmonary sequelae. OBJECTIVE To evaluate the distribution of whole blood T cell subsets, serum cytokines, and biomarkers of pulmonary fibrosis in PCM patients, according to the clinical form and at different time points, during the antifungal therapy. METHODS Eighty-seven PCM patients, from an endemic area in Brazil, were categorised into groups, according to the clinical form (AF or CF) and the moment of treatment. The peripheral blood T lymphocyte subsets of these patients were analysed using fluorescence-activated cell sorting. The serum levels of cytokines, basic fibroblast growth factor and surfactant protein-D (SP-D) were also analysed. FINDINGS In the CF patients, an expansion of the peripheral blood TCD4+ cells was observed during the treatment, and this persisted even after two years of antifungal treatment. In addition, these patients showed high serum levels of SP-D. CONCLUSION Our findings highlight the immunological changes CF patients undergo, during and after treatment, possibly due to the hypoxia triggered by pulmonary fibrosis and emphysema.

Abstract Purpose: To investigate the effects of cyclosporine A on renal ischemia-reperfusion injury during transient hyperglycemia in rats. Methods: In a model of ischemia-reperfusion-induced renal injury and transiently induced hyperglycemia by intraperitoneal injection of glucose, 2.5 g.kg-1, Wistar rats were anesthetized with either isoflurane or propofol and received intravenous cyclosporine A, 5 mg.kg-1, five minutes before reperfusion. Comparison groups were isoflurane and propofol sham groups and isoflurane and propofol ischemia-reperfusion-induced renal injury. Renal tubular cell viability was quantitatively assessed by flow cytometry after cell culture and classified as early apoptosis, necrotic cells, and intact cells. Results: Early apoptosis was significantly higher in isoflurane and propofol anesthetized animals subjected to renal ischemia-reperfusion injury when compared to both cyclosporine A treated and sham groups. Necrosis percentage was significantly higher in propofol-anesthetized animals subjected to renal ischemia-reperfusion injury. The percentage of intact cells was lower in both, isoflurane and propofol anesthetized animals subjected to renal ischemia-reperfusion injury. Conclusion: In a model of ischemia-reperfusion-induced renal injury, cyclosporine A, 5 m.kg-1, administered five minutes before renal reperfusion in rats with acute-induced hyperglycemia under either isoflurano or propofol anesthesia, attenuated early apoptosis and preserved viability in renal tubular cells, regardless of the anesthetic used.

ABSTRACT PURPOSE: To evaluate the effects of dobutamine (DB), noradrenaline (NA), and their combination (NADB), on volume retention in rabbits submitted to hemorrhage. METHODS: Thirty six rabbits were randomly divided into 6 groups: SHAM, Control, Saline, DB, NA, DB+NA. All the animals, except for SHAM, were subjected to hemorrhage of 25% of the calculated blood volume. Control animals were replaced with their own blood. The other groups received NSS 3 times the volume withdrawn. The intravascular retention, hematocrit, diuresis, central venous pressure, mean arterial pressure, NGAL, dry-to-wet lung weight ratio (DTWR) and the lung and kidney histology were analyzed. RESULTS: Replacement with NSS and NA, DB or NA+DB did not produce differences in the intravascular retention. After hemorrhage, the animals presented a significant decrease in the MAP and CVP, which were maintained until volume replacement. Regarding NGAL, dry-to-wet-lung-weight ratio, lung and kidney histology, there were no statistical differences between the groups. CONCLUSION: The use of noradrenaline, dobutamine or their combination did not increase the intravascular retention of volume after normal saline infusion.

ABSTRACT PURPOSE: To investigate changes in the serum concentration and renal expression of IL-1 and TNF-α cytokines in rats that received sevoflurane and glibenclamide prior to hemorrhage. METHODS: Two groups of sevoflurane-anesthetized Wistar rats (n=10): G1 (control) and G2 (glibenclamide, 1 µg/g i.v.); hemorrhage of 30% blood volume (10% every 10 min), with replacement using Ringer solution, 5 ml/kg/h. Serum concentrations of IL-1 and TNF-α were studied in the first hemorrhage (T1) and 50 min later (T2), renal expression, at T2. RESULTS: In serum, G1 TNF-α (pg/mL) was T1=178.6±33.5, T2=509.2±118.8 (p<0.05); IL-1 (pg/mL) was T1=148.8±31.3, T2=322.6±115.4 (p<0.05); in G2, TNF-α was T1=486.2±83.6, T2=261.8±79.5 (p<0.05); IL-1 was T1=347.0±72.0, T2= 327.3±90.9 (p>0.05). The expression of TNF-α and IL-1 in the glomerular and tubular cells was significantly higher in the G2 group. CONCLUSIONS: Hemorrhage and glibenclamide elevated TNF-α and IL-1 concentrations in serum and kidneys. High levels of TNF-α already present before the hemorrhage in the glibenclamide group may have attenuated the damages found in the kidneys after the ischemia event.

Growth functions with inflection points following a diphasic model, can be adjusted by two approaches using segmented regression or the sum of two functions. In both cases, there are two functions, one for each phase, with inflection and stability points. However, when they are summed, the result is a new function and the points of inflection and stability are different from those obtained from using each function individually. A method to determine these points in a diphasic logistics sum of functions is suggested and the results obtained from fitting the models to eucalyptus growth data showed a better fit of the logistic diphasic sum as compared with segmented regression and monophasic logistic models.

O objetivo deste estudo foi analisar e comparar os registros eletrocardiográficos de 50 equinos American Miniature Horse, clinicamente saudáveis, obtidos pelos métodos convencional e computadorizado, para a padronização do método informatizado. As medidas de duração e amplitude das ondas e intervalos e frequência cardíaca foram analisadas nas derivações do plano frontal e na derivação base-ápice. Diferenças quanto à duração da onda P, foram encontradas nas derivações I e aVL, e do complexo QRS, em todas as derivações do plano frontal. Quanto à derivação base-ápice, houve diferença na duração do intervalo QT e da onda T, assim como na amplitude da onda S. Concluiu-se, portanto, que ocorre superestimação da duração do complexo QRS no método computadorizado e que, devido às diferenças inerentes de cada método, torna-se importante a utilização de valores de referência para o método computadorizado também na espécie equina

The aim of this study was to analyze and compare electrocardiographic records of 50 horses American Miniature Horse, clinically healthy, obtained by conventional methods and computerized, to the standardization of the computerized method. The measures of duration and amplitude of waves and intervals and heart rate were analyzed in the leads of the frontal plane and base-apical lead. Differences about the duration of the P wave were found in I and aVL derivations, and QRS complex, in all derivations of the frontal plane. As for basis-apical derivation, there was no difference in the duration of the QT interval and the T wave and the amplitude of the S wave, therefore concluding that overestimation occurs in the duration of the QRS complex in computerized method and that due to inherent differences of each method, it becomes important the use of reference values for the computerized method also in the equine species

Adjusting autoregressive and mixed models to growth data fits discontinuous functions, which makes it difficult to determine critical points. In this study we propose a new approach to determine the critical stability point of cattle growth using a first-order autoregressive model and a mixed model with random asymptote, using the deterministic portion of the models. Three functions were compared: logistic, Gompertz, and Richards. The Richards autoregressive model yielded the best fit, but the critical growth values were adjusted very early, and for this purpose the Gompertz model was more appropriate.

Mice experimentally infected with a pathogenic strain of Leptospira interrogans serovar Canicola produced false negative results (prozone effect) in a microscopic agglutination test (MAT). This prozone effect occurred in several serum samples collected at different post-infection times, but it was more prominent in samples collected from seven-42 days post-infection and for 1:50 and 1:100 sample dilutions. This phenomenon was correlated with increased antibody titres in the early post-infection phase. While prozone effects are often observed in serological agglutination assays for the diagnosis of animal brucellosis and human syphilis, they are not widely reported in leptospirosis MATs.

PURPOSE: To study the effect of isoflurane (Iso) or propofol (Prop) anesthesia on renal ischemia/reperfusion injury (IRI) during transient hyperglycemia. METHODS: Thirty six rats were randomly assigned into six groups of six animals each: PHS (Sham-Prop=1mg.kg-1.min-1 + Hyperglycemia=2.5g.kg-1 of glucose solution administered intraperitoneally); HIS (Sham-Iso + Hyperglycemia); PHI (Prop + Hyperglycemia + Ischemia); IHI (Iso + Hyperglycemia + Ischemia); PI (Prop + Ischemia), and II (Iso + Ischemia). After 30 minutes of anesthesia induction, right nephrectomy was performed (all animals) and the left renal artery was clamped during 25 minutes (ischemia). The animals were sacrificed after 24 hours and blood collection (to dose creatinine) and left kidney removal were performed for histological analysis, and flow cytometry (FCM): percentage of initial apoptosis (APTi) and viable cells (VC). RESULTS: Serum creatinine (mg/dL) was statistically different in groups PHI (3.60±0.40) and IHI (3.23±1.08), p<0.05. Histological analysis was statistically different in groups PHI (4.0[4.0;5.0]) and IHI (4.5[4.0;5.0]), p<0.05. APTi percentage was statistically different in groups PHI (73.2±7.1), and IHI (48.1±14). VC percentage was statistically different in groups PHI (25.8±6.9) and IHI (38.5±9.2), p<0.05. CONCLUSIONS: Propofol and isoflurane showed the same level of protection against ischemia/reperfusion injury in the normoglycemic groups. Transient hyperglycemia is associated with an increase in IRI.