ABSTRACT Serodiagnosis of strongyloidiasis is usually performed by ELISA for the detection of IgG antibodies due to its high sensitivity and practicality, but its main limitation is a constant source of S. stercoralis antigens. The use of S. venezuelensis as a heterologous source of antigens has facilitated several published studies on the serodiagnosis and epidemiology of human strongyloidiasis. The main objective of this study was to evaluate the diagnostic accuracy of surface cuticle antigens of infective larvae of S. venezuelensis extracted with CTAB detergent (L3-CTAB) in comparison with soluble somatic extracts (L3-SSE) using a panel of sera from immunocompetent and immunocompromised individuals, at three different cut-offs. ROC curve analysis showed that L3-CTAB had an AUC of 0.9926. At the first cut-off value (OD 450 nm = 0.214), sensitivity and specificity were 100% and 90.11%, respectively, with a diagnostic accuracy of 0.93. At a second cut-off value (OD 450 nm = 0.286), sensitivity and specificity were 70% and 100%, respectively, with a diagnostic accuracy of 0.91. However, at an alternative third cut-off value (OD 450 nm = 0.589), sensitivity and specificity were 95% and 97.8%, respectively, with a diagnostic accuracy of 0.97. Using L3-CTAB as an antigenic source, the seropositivity rate in immunocompromised patients was 28.13% (9/32) whereas a seropositivity rate of 34.38% (11/32) was found when L3-SSE was used in ELISA. Therefore, the L3-CTAB is simple and practical to obtain and was found to be highly sensitive and specific. practicality S L3CTAB LCTAB L3 L (L3-CTAB L3SSE LSSE SSE (L3-SSE individuals cutoffs. cutoffs cut offs. offs cut-offs 09926 0 9926 0.9926 cutoff off OD 45 0.214, 0214 0.214 , 214 0.214) 100 9011 90 11 90.11% respectively 093 93 0.93 0.286, 0286 0.286 286 0.286) 70 091 91 0.91 However 0.589, 0589 0.589 589 0.589) 95 978 97 8 97.8% 097 0.97 2813 28 13 28.13 9/32 932 9 32 (9/32 3438 34 38 34.38 11/32 1132 (11/32 Therefore specific 0992 992 0.992 4 021 0.21 21 10 901 1 90.11 09 0.9 028 0.28 7 058 0.58 58 97.8 281 2 28.1 9/3 3 (9/3 343 34.3 11/3 113 (11/3 099 99 0.99 02 0.2 90.1 0. 05 0.5 5 97. 28. 9/ (9/ 34. 11/ (11/ 90. (9 (11 ( (1

Abstract Objective: The association between diabetes and Strongyloides infection remains controversial. This study aimed to detect Strongyloides stercoralis DNA in the feces of patients with Diabetes Mellitus type 2 (DM2). Methods: Fecal samples were analyzed via the Lutz, Rugai, and agar plate culture methods. PCR amplification was performed using two targets (PCR-genus and PCR-species) located on the S. stercoralis 18S ribosomal. Results: The positivity for S. stercoralis using parasitological methods was 1.1%. PCR-genus (14.13%) demonstrated a higher positivity than PCR-species (9.78%). Conclusion: The results confirm the greater positivity of the molecular diagnosis in relation to parasitological methods, reinforcing its use as an additional tool for the diagnosis of S. stercoralis infection in patients with DM2 living in endemic areas for this helminthiasis. HIGHLIGHTS Positivity for strongyloidiasis in coproscopic exam was low in diabetic patients. PCR is more sensitive for detecting S. stercoralis infection in diabetic patients. Molecular diagnosis is an important tool for the detection of S. stercoralis.

ABSTRACT The Western-blotting technique was applied to identify antigenic fractions of excretory-secretory Toxocara canis antigen recognized by IgG antibodies throughout an experimental infection in mice challenged by different inocula. Mice were inoculated with 5, 50 and 500 embryonated eggs and serum samples were collected 15, 30, 60, 90 and 120 days post-infection. Serum samples were analyzed using an excretory-secretory Toxocara antigen. Antibodies recognized antigenic fractions from 30 to 90 kDa. The protein fraction of 30-35 kDa was the most frequently recognized regardless of the size of inoculum and the stage of infection represented by the different collection times, but the antigenic recognition was more evident in groups infected with 50 and 500 eggs. This study presents an antigenic panel of the excretory-secretory antigen of T. canis and suggests that the 30-35 kDa antigenic fraction is a promising marker of the infection and should be further explored in future studies on experimental toxocariasis.

ABSTRACT This study aimed to report the first case of a patient with hepatosplenic schistosomiasis mansoni, refractory ascites and portal vein thrombosis treated with a transjugular intrahepatic portosystemic shunt (TIPS), at the Instituto de Radiologia, Hospital das Clinicas, Faculdade de Medicina, Universidade de Sao Paulo, Brazil. After the procedure, the patient recovered favorably and progressed with portal pressure reduction and no deterioration of the liver function. Endovascular shunt modification is a conservative medical approach that often helps in reducing symptoms significantly, making it a less invasive and a safer alternative to liver transplantation for the treatment of schistosomiasis with portal hypertension.

ABSTRACT Blastocystis sp. is an enteric protist commonly found in human fecal samples. In Brazil, few studies have been developed, but none of them has explored the presence of Blastocystis in patients with diabetes mellitus. We evaluated the occurrence and molecular identification of Blastocystis sp. among patients with diabetes mellitus in the Midwest region, Goias State, Brazil. Genomic DNA was obtained from 175 fecal samples (99 from the diabetic group and 76 from the control group). PCR was performed using pan-Blastocystis primers from the SSU-rDNA gene. Microscopic examination revealed positivity of 12.1% and 7.9% for Blastocystis in diabetics and in controls, respectively. Amplification of Blastocystis DNA was observed in 34.4% (34 of 99) and 30.3% (23 of 76) from the diabetic and control groups, respectively. Phylogenetic analyses and BLAST searches revealed six subtypes among Blastocystis isolates in the diabetic group, represented by ST1 (38.2%), ST2 (11.8%), ST3 (35.3%), ST6 (2.9%), ST7 (2.9%) and ST8 (8.8%). In the control group, ST1 (21.8%), ST2 (21.8%), ST3 (43.5%), ST6 (4.4%) and ST8 (8.7%) were identified. This study is the first report regarding the occurrence and subtypes distribution of Blastocystis in patients with diabetes mellitus in Brazil. The results reinforce the potential risk of Blastocystis infection in patients with diabetes, in addition, it contributes to the understanding of the genetic diversity of this enigmatic organism.

Resumo: Introdução: Uma extensa literatura demonstra os benefícios do ensino ativo na educação médica e a necessidade de atualização das metodologias de ensino. As estratégias de aprendizagem ativa consolidam o conhecimento teórico e estimulam as habilidades conceituais do aluno. No entanto, reformar a prática pedagógica é desafiador. Existem diversas estratégias de aprendizagem ativa que sugerem uma adaptação do formato de entrega do conteúdo, de modo a adequá-las a diversos contextos. Neste artigo, discutimos a experiência da aplicação da técnica Four Corners no ensino de doenças infecciosas em nossa instituição. Relato de experiência: A estratégia Four Corners foi implementada da seguinte maneira: 1. preparação: os estudantes recebem material de estudo para análise prévia; 2. preparação da sala: cada um dos quatro cantos recebe cartelas com os casos clínicos escolhidos; 3. divisão de tarefas: as tarefas são divididas entre os estudantes; 4. atividade: os estudantes leem e discutem cada um dos casos propostos com mínima interferência do moderador; 5. finalização: discussão final com todos os participantes liderados pelo moderador. A duração de toda a tarefa varia de duas horas e meia a três horas. Discussão: O processo educacional emerge da experiência do aprendiz. Muitos professores do ensino superior acreditam que promovem o pensamento crítico por meio do ensino ativo em suas aulas, entretanto a porcentagem dos que aplicam essas técnicas ainda é baixa. Existe uma ampla variedade de técnicas ativas de ensino, e muitas delas se adaptam a diferentes cenários. A técnica Four Corners é uma estratégia de aprendizagem ativa que promove debate e troca de conceitos entre os estudantes. Conclusão: Este relato de experiência descreve uma estratégia de aprendizagem ativa para o ensino de doenças infecciosas para estudantes de Medicina. O aprimoramento da educação médica passa por entender o papel do professor como mediador do processo de aprendizagem. Propor novas estratégias de ensino é desafiador e requer ajustes frequentes. A estratégia Four Corners melhora o envolvimento do aluno com o processo de aprendizagem e é eficiente para compreender um conteúdo teórico extenso em um tempo de discussão relativamente curto.

Abstract: Introduction: An extensive literature has demonstrated the benefits of active learning in medical education and has revealed the need for updating the teaching methodologies. Active learning strategies consolidate the theoretical knowledge, while simultaneously encouraging the student´s concrete abilities. Nonetheless, remodeling the pedagogical practice is challenging. Several active learning strategies are available and selecting the most suitable one is often a matter of format, not of content. In this article, we discuss the experience of the ‘Four Corners’ technique in the teaching of Infectious Diseases during the internship at our institution. Experience report: The four corners strategy can be adapted to different topics and should be implemented as follows: 1. pre-activity learning: students receive the supportive material for previous analysis; 2. Preparation of the room: each of the four corners is labeled with a different case vignette; 3. Dividing tasks: the specific assignments are divided among the students; 4. Activity: the students read the case vignette and discuss the questions, while the mediator has an observer’s role, and 5. Debriefing: final discussion with all participants led by the mediator. The entire process takes from 2h30 to 3 hours. Discussion: The educational process emerges from the learner´s experience. Many instructors working in higher education believe they promote critical thinking and active teaching during their classes; however, the percentage of teachers who regularly use active teaching strategies is still low. A range of different active learning strategies have been described and many can be adapted to different scenarios; most approaches change the nature of the learning experience but do not always require a change in the given subject. The Four Corners technique is an active learning strategy that promotes debate and exchange of ideas among students. Conclusion: This experience report describes an active learning strategy for the teaching of Infectious Diseases to medical students. The medical education improvement goes through highlighting the teacher’s role as a mediator of the learning process; proposing new teaching strategies is challenging and requires frequent adjustments. The Four Corners strategy improves student’s engagement with the learning process, and it is an efficient strategy to comprise an extensive theoretical content in a relatively short discussion time.

The present study aimed to evaluate the occurrence of Blastocystis sp. in Brazilian studies over a period of years (2000-2020), as well as point out relevant aspects of this enigmatic organism. We performed a literature search using six sources of international databases. The data were divided into diagnostic by parasitological and molecular techniques, and relevant aspects. After applying the inclusion and exclusion criteria, 52 studies were included in the final analysis. The occurrence of Blastocystis sp. in Brazil ranged from 0.5% to 86.6%, as determined using parasitological techniques. The highest occurrence was in the North (27.3%) and the lowest, in the Midwest region (13.4%). In Brazil, most studies have employed molecular techniques and are concentrated in the Southeast region. The Blastocystis sp. subtype ST3 had the highest average positivity, followed by ST1 and ST2. These findings represent a panorama that reflects the reality of Brazil; thus, we believe that the effectiveness of parasitological diagnosis should be considered with regard to making an appropriate choice of technique for detecting Blastocystis sp. Additionally, we emphasize the importance of further studies in the context of molecular epidemiology with regard to this genus. Blastocystis sp. is not well understood yet, and very little information regarding this genus is available; hence, further research regarding this genus is urgently needed.

ABSTRACT Approximately 240 million people worldwide are infected by Schistosoma. In Brazil, one of the main intermediate hosts of this parasite is Biomphalaria glabrata snails. The early detection of larval stages in intermediate hosts is an important challenge to public health, but it also represents an opportunity as a new alternative to indicate earlier natural infections before cercariae differentiation and emergence. In this context, we demonstrated that PCR amplification of a 28S gene fragment from the parasite does demonstrate S. mansoni infection in snails 14 days post infection. This conventional polymerase chain reaction amplified clear bands and was able to detect parasitic infection in the intermediate host B. glabrata under experimental conditions. However, we reinforce that this approach requires deeper investigations and further comparisons to confirm its specificity and sensitivity in earlier time points after miracidia infection. This approach has relevant potential as an effective molecular-based strategy for the monitoring of schistosomiasis transmission.

ABSTRACT In experimental infection with Strongyloides venezuelensis, the acute and recovery phases can be distinguished, unlike human infections caused by Strongyloides stercoralis. The objective of this study was to evaluate the production of anti-Strongyloides IgG antibodies and the recognition of immunogenic protein bands during the acute and the recovery phases in rats experimentally infected with S. venezuelensis. Rats were infected subcutaneously with 400 or 4,000 S. venezuelensis infective larvae. The acute phase was characterized by elimination of a large number of eggs in the faeces on days 6-14 post infection; the recovery phase was characterized by the resolution of the infection between days 30 and 35 post infection. Differences in IgG levels were observed in the acute and the recovery phases. Different antigenic fractions were recognized in both phases of infection. It is concluded that proteins within the 30-40 kDa range are immunoreactive markers for both the acute and the recovery phases in rats experimentally infected with S. venezuelensis, particularly using membrane antigen.

ABSTRACT Blastocystis sp. is a protist commonly found in stool samples of humans and animals. Biological and genetic factors of this organism remain controversial. The present study aimed to develop and implement the Blastocystis in vitro culture of Brazilian human isolates for routine use. The fecal isolates (n = 20) were maintained in our laboratory by several passages in Pavlova's medium. Cultures were monitored every 72 h by light microscopy. Genomic DNA was extracted to identify the subtypes (STs). In most isolates, the vacuolar form was prevalent. The amoeboid, granular and cystic forms were observed during in vitro cultivation. STs 1, 2, 3, 4 and 7 were identified. Our preliminary results show the generation time and forms present in the in vitro culture of Blastocystis subtypes isolated from Brazilian human isolates. Therefore, we emphasize the use of in vitro culture as a tool in future studies for the better understanding of the biological aspects of Blastocystis sp.

OBJECTIVES: Hyperinfection or disseminated strongyloidiasis has been frequently reported after transplants and is related to high mortality. This study aimed to screen for strongyloidiasis using serological diagnoses in transplant candidates. METHODS: An ELISA test was performed with filariform larvae of Strongyloides venezuelensis as a source of antigen. RESULTS: In the serum from transplant candidates, anti-Strongyloides IgG antibodies were detected in 35/150 (23.3%) samples by soluble fractions in phosphate buffered saline (PBS), 31/150 (20.7%) samples by soluble fractions in Tris-HCl, 27/150 (18.0%) samples by membrane fractions in PBS and 22/150 (14.7%) samples by membrane fractions in Tris-HCl. CONCLUSIONS: The present results suggest the ELISA test, ideally using soluble fractions of filariform larvae S. venezuelensis in PBS, as an additional strategy for the diagnosis of strongyloidiasis in transplant candidates.

OBJECTIVE: Schistosomiasis remains a public health problem. This was a descriptive and retrospective study of 42 patients with a severe form of schistosomiasis who were admitted to the outpatient clinic of the Faculdade de Medicina da Universidade de São Paulo, São Paulo, in São Paulo, Brazil. METHODS: A data collection questionnaire was designed from the patient charts, and the following variables were evaluated: age, sex, place of birth, occupation, signs and symptoms of schistosomiasis, data from laboratory and imaging examinations, data regarding treatment outcomes, and the existence of comorbidities. Statistical analysis was carried out using Statistical Package for the Social Sciences 15.0 and Microsoft Excel 2003 software. The significance levels of the tests were fixed, accepting 5% type 1 error (α=0.05). Since this was a retrospective observational study, not all data were available for analysis. RESULTS: The mean age of the patients was 48.24 years; 57.1% were male. Statistically significant associations were observed between splenomegaly and thrombocytopenia (p=0.004) and between splenomegaly and leukopenia (p=0.046); however, only 4.5% of the patients had esophageal hemorrhage. Some patients received a specific treatment; of those, 42% took praziquantel, and 35.4% took oxamniquine. Nonspecific drug therapy was given as follows: 65% received propranolol, 90% omeprazole, and 43.6% aluminum hydroxide. The other treatments were as follows: 92.9% of patients underwent endoscopic treatment, 85% received sclerotherapy, and 62.5% used elastic bandages. CONCLUSION: This preliminary study presents a multidisciplinary outpatient follow-up associated with endoscopic and drug treatments that may be effective at preventing bleeding.