Abstract The enterotoxigenic Escherichia coli (ETEC) strain is one of the most frequent causative agents of childhood diarrhea and travelers’ diarrhea in low-and middle-income countries. Among the virulence factors secreted by ETEC, the exoprotein EtpA has been described as an important. In the present study, a new detection tool for enterotoxigenic E. coli bacteria using the EtpA protein was developed. Initially, antigenic sequences of the EtpA protein were selected via in silico prediction. A chimeric recombinant protein, corresponding to the selected regions, was expressed in an E. coli host, purified and used for the immunization of mice. The specific recognition of anti-EtpA IgG antibodies generated was evaluated using flow cytometry. The tests demonstrated that the antibodiesdeveloped were able to recognize the native EtpA protein. By coupling these antibodies to magnetic beads for the capture and detection of ETEC isolates, cytometric analyses showed an increase in sensitivity, specificity and the effectiveness of the method of separation and detection of these pathogens. This is the first report of the use of this methodology for ETEC separation. Future trials may indicate their potential use for isolating these and other pathogens in clinical samples, thus accelerating the diagnosis and treatment of diseases. (ETEC travelers lowand low middleincome middle income countries important study E developed Initially prediction regions host mice antiEtpA anti cytometry isolates sensitivity samples diseases

Abstract: Introduction: Communication skills (CS) are indispensable competencies for any healthcare professional as such skills lead to greater patient compliance, understanding, and overall satisfaction. The constant aging of the Brazilian population increasingly results in the presence of the older people in the healthcare system. This has led medical schools to think about a reformulation of their curricular components within the scope of CS, since this subject still presents gaps. In this context, intergenerational programs play a valuable role in the development of CS by allowing interaction between different generations. Objective: To investigate the perspectives of health students about their participation in an intergenerational program in the city of Paulo Afonso/BA entitled “[Tec-Idoso]: utilização de tecnologia como ferramenta de inclusão digital e de apoio psicossocial ao idoso” [Tec-Idoso: use of technology as a tool for digital including and psychosocial support for the elderly], linked to the Federal University of Vale of São Francisco (UNIVASF). Method: This is an exploratory qualitative study based on semi-structured interviews, using the data analysis method. Results: The following thematic categories were defined: intergenerational contact, perception and stigma about old age, CS, impact on academic performance and impact on future professional practices. Conclusion: Given the above, the results obtained from this research include improved academic performance and social interaction through the development of CS, change of paradigms facing aging, and preparation for handling personal and professional life situation with more confidence. Abstract Introduction (CS compliance understanding satisfaction system gaps context generations Objective AfonsoBA Afonso BA TecIdoso Tec Idoso “[Tec-Idoso] idoso [Tec-Idoso elderly, elderly , elderly] UNIVASF. UNIVASF . (UNIVASF) Method semistructured semi structured interviews method Results defined contact age practices Conclusion above confidence “[Tec-Idoso (UNIVASF

Resumo: Introdução: As habilidades de comunicação (HC) são competências indispensáveis para qualquer profissional da área de saúde, pois permitem maior adesão, compreensão e satisfação geral do paciente. O constante envelhecimento da população brasileira resulta cada vez mais na presença do público idoso no sistema de saúde. Esse cenário tem levado as escolas médicas a pensar em uma reformulação dos seus componentes curriculares no âmbito das HC, visto que esse assunto ainda apresenta lacunas. Nesse contexto, os programas intergeracionais possuem um valioso papel no desenvolvimento das HC por permitirem a interação entre diferentes gerações. Objetivo: Este estudo teve como objetivo investigar as perspectivas de estudantes da área de saúde sobre a participação em um programa intergeracional no município de Paulo Afonso, na Bahia, intitulado “[Tec-Idoso]: utilização de tecnologia como ferramenta de inclusão digital e de apoio psicossocial ao idoso”, vinculado à Universidade Federal do Vale do São Francisco (Univasf). Método: Trata-se de um estudo qualitativo e exploratório, realizado a partir de entrevistas semiestruturadas, utilizando o método de análise de conteúdo. Resultado: As seguintes categorias temáticas foram definidas: contato intergeracional, percepção e estigma com pessoas idosas, HC, impacto no desempenho acadêmico e impacto nas práticas profissionais futuras. Conclusão: Os resultados da presente pesquisa incluem melhora do desempenho acadêmico e da interação social por meio do desenvolvimento das HC, mudança de perspectivas perante o envelhecimento e preparação para o enfrentamento com mais confiança de situações da vida pessoal e profissional. Resumo Introdução (HC adesão paciente lacunas contexto gerações Objetivo Afonso Bahia TecIdoso Tec Idoso “[Tec-Idoso] idoso, , idoso” Univasf. Univasf . (Univasf) Método Tratase Trata se exploratório semiestruturadas conteúdo Resultado definidas idosas futuras Conclusão “[Tec-Idoso (Univasf

Respiratory failure (RF) is the main cause of hospital admission in HIV/AIDS patients. This study assessed comorbidities and laboratory parameters in HIV/AIDS inpatients with RF (N = 58) in relation to those without RF (N = 36). Tuberculosis showed a huge relative risk and platelet counts were slightly higher in HIV/AIDS inpatients with RF. A flow cytometry assay for reactive oxygen species (ROS) showed lower levels in platelets of these patients in relation to the healthy subjects. However, when stimulated with adrenaline, ROS levels increased in platelets and platelet-derived microparticles of HIV/AIDS inpatients, which may increase the risk of RF during HIV and tuberculosis (HIV-TB) coinfection.

BACKGROUND Thrombocytopenia in malaria involves platelet destruction and consumption; however, the cellular response underlying this phenomenon has still not been elucidated. OBJECTIVE To find associations between platelet indices and unbalanced Th1/Th2/Th17 cytokines as a response to thrombocytopenia in Plasmodium vivax infected (Pv-MAL) patients. METHODS Platelet counts and quantification of Th1/Th2/Th17 cytokine levels were compared in 77 patients with uncomplicated P. vivax malaria and 37 healthy donors from the same area (endemic control group - ENCG). FINDINGS Thrombocytopenia was the main manifestation in 55 patients, but was not associated with parasitaemia. The Pv-MAL patients showed increases in the mean platelet volume (MPV), which may be consistent with larger or megaplatelets. Contrary to the findings regarding the endemic control group, MPV and platelet distribution width (PDW) did not show an inverse correlation, due the increase in the heterogeneity of platelet width. In addition, the Pv-MAL patients presented increased IL-1β and reduced IL-12p70 and IL-2 serum concentrations. Furthermore, the reduction of these cytokines was associated with PDW values. MAIN CONCLUSIONS Our data demonstrate that an increase in MPV and the association between reductions of IL-2 and IL-12 and PDW values may be an immune response to thrombocytopenia in uncomplicated P. vivax malaria.

As phagocytosis is the first line of defense against malaria, we developed a phagocytosis assay with Plasmodium vivax (P. vivax) merozoites that can be applied to evaluate vaccine candidates. Briefly, after leukocyte removal with loosely packed cellulose powder in a syringe, P. vivax trophozoites matured to the merozoite-rich schizont stages in the presence of the E64 protease inhibitor. The Percoll gradient-enriched schizonts were chemically disrupted to release merozoites that were submitted to merozoite opsonin-dependent phagocytosis in two phagocytic lines with human and mouse antibodies against the N- and C-terminus of P. vivax Merozoite Surface Protein-1 (Nterm-PvMSP1 and MSP119). The resulting assay is simple and efficient for use as a routine phagocytic assay for the evaluation of merozoite stage vaccine candidates.

ABSTRACT Objectives The increase in HIV transmissibility in non-ulcerative sexually transmitted infection is already well-established. It is estimated that symptomatic carriers of N. gonorrhoeae and C. trachomatis have a relative risk of 4.8-fold and 3.6-fold, respectively, for the sexual acquisition of HIV. This type of evaluation for asymptomatic urethritis is necessary to reinforce strategies to combat HIV transmission. This study aims to assess the prevalence of patients with asymptomatic urethritis among men diagnosed with HIV-1 and determine the risk factors associated with this infection. Methods We enrolled a total of 115 male patients aged 18 years or older who have been diagnosed with HIV infection and have no symptoms of urethritis or other sexually transmitted infections and who have been evaluated between May and August 2015 in a follow-up visit at the Immunology Outpatient Clinic of a Brazilian University Hospital. Results Four asymptomatic patients were positive for C. trachomatis and were considered asymptomatic carriers of urethritis. Prevalence was 3.47%. Patients who were positive for C. trachomatis urethritis had a lower mean age (p = 0.015). Conclusion The presence of asymptomatic sexually transmitted infection is a challenge in clinical practice. We recommend that, in outpatient practice, the habit of inquiring on previous sexual behavior to obtain more information about risks and associations with asymptomatic sexually transmitted infection, a routine physical examination and complementary tests to detect STI pathogens should be performed to discard these conditions. The development of rapid tests for this purpose should also be encouraged.

ABSTRACT A prevalence of 3.47% of asymptomatic Chlamydia trachomatis urethritis has been previously reported among males living with HIV infection in Brazil. This study aims to assess the recurrence of C. trachomatis urethritis three years later in the same cohort of patients and analyze associated risk factors. A total of 115 male patients diagnosed with HIV infection, with no symptoms of urethritis and observed since May of 2015 in followup visits were enrolled. They had urine samplers tested by PCR for C. trachomatis and N. gonorrhoeae between February and March 2018. Results: Three of the four patients who had asymptomatic C. trachomatis urethritis three years before were recurrently positive for C. trachomatis urethritis. Two new patients were diagnosed as positives, accounting for a total asymptomatic C. trachomatis urethritis prevalence of 4.34%. The prevalence during the whole study was 5.21%. The relative risk for a new urethritis episode among those previously diagnosed with urethritis is RR=41.62 (95% CI: 9.42-183.84), p < 0.01. Patients who presented asymptomatic urethritis anytime and who were recurrently positive for C. trachomatis had a lower mean age (p<0.01). Married individuals were protected regarding asymptomatic urethritis [p<0.01, OR = 0.04 (0.005-0.4)] and had lower risk to develop recurrence [p<0.01, RR = 0.86 (0.74-0.99)]. Illicit drugs users had risk associated to asymptomatic urethritis [p=0.02, OR= 5.9 (1.03-34)] and higher risk to develop recurrence [p<0.01, RR=1.1 (1-1.22)]. Conclusion: The recurrence of asymptomatic C. trachomatis urethritis after treatment among males living with HIV infection in Brazil can be considered high and should not be neglected.

A consolidação da Atenção Primária à Saúde (APS) requer políticas públicas embasadas por evidências científicas. Este artigo apresenta o estudo ELECT, cujo objetivo foi identificar temas prioritários de pesquisa para a fortalecimento da APS no estado de São Paulo, Brasil. Com a participação de especialistas e de um grupo focal com usuários, foi obtida uma lista com os vinte principais obstáculos, bem como dez temas de pesquisa prioritários, na APS. Os resultados apontam para problemas e temas de pesquisas relacionados à: organização da gestão, capacitação de profissionais e gestores, valorização profissional, criação de mecanismos de colaboração entre equipes de saúde e informatização dos recursos. Espera-se, assim, estimular o debate no contexto da APS sobre o papel da priorização de pesquisas, seus obstáculos e proposições de pesquisa. Almeja-se, também, estimular a adoção de modelos mais participativos de seleção de temas de pesquisa.

The consolidation of Primary Health Care (PHC) requires public policies based on scientific evidence. This paper presents the ELECT study, aimed to identify priority research themes for strengthening PHC in the state of Sao Paulo, Brazil. A list of the twenty main obstacles and ten priority research themes in PHC were obtained with participation of specialists and a focus group with users. The results point to problems and research issues related to organizational management, training of professionals and managers, professional development, creation of cooperation mechanisms between health teams and computerization of resources. It is expected to stimulate debate in the context of the PHC on the role of research prioritization, its obstacles and research propositions. It also aims to encourage the adoption of more participatory models of selection of research topics.

La solidificación de la Atención Primaria de Salud (APS) requiere políticas públicas con base en evidencias científicas. Este artículo presenta el estudio ELECT, cuyo objetivo fue identificar temas prioritarios de investigación para el fortalecimiento de la APS en el estado de São Paulo. Con la participación de especialistas y de un grupo de opinión formado por usuarios, se obtuvo una lista con los veinte principales obstáculos, así como diez temas de investigación prioritarios en la APS. Los resultados señalan problemas y temas de investigación relacionados a la organización de la gestión, la capacitación de profesionales y gestores, la valorización profesional, la creación de mecanismos de colaboración entre equipos de salud e informatización de los recursos. Se espera por lo tanto incentivar el debate en el contexto de la APS sobre el papel de la priorización de investigaciones, sus obstáculos y propuestas de investigación. Se anhela también incentivar la adopción de modelos más participativos de selección de temas de investigación.

A consolidação da Atenção Primária à Saúde (APS) requer políticas públicas embasadas por evidências científicas. Este artigo apresenta o estudo ELECT, cujo objetivo foi identificar temas prioritários de pesquisa para a fortalecimento da APS no estado de São Paulo, Brasil. Com a participação de especialistas e de um grupo focal com usuários, foi obtida uma lista com os vinte principais obstáculos, bem como dez temas de pesquisa prioritários, na APS. Os resultados apontam para problemas e temas de pesquisas relacionados à: organização da gestão, capacitação de profissionais e gestores, valorização profissional, criação de mecanismos de colaboração entre equipes de saúde e informatização dos recursos. Espera-se, assim, estimular o debate no contexto da APS sobre o papel da priorização de pesquisas, seus obstáculos e proposições de pesquisa. Almeja-se, também, estimular a adoção de modelos mais participativos de seleção de temas de pesquisa.

The consolidation of Primary Health Care (PHC) requires public policies based on scientific evidence. This paper presents the ELECT study, aimed to identify priority research themes for strengthening PHC in the state of Sao Paulo, Brazil. A list of the twenty main obstacles and ten priority research themes in PHC were obtained with participation of specialists and a focus group with users. The results point to problems and research issues related to organizational management, training of professionals and managers, professional development, creation of cooperation mechanisms between health teams and computerization of resources. It is expected to stimulate debate in the context of the PHC on the role of research prioritization, its obstacles and research propositions. It also aims to encourage the adoption of more participatory models of selection of research topics.

La solidificación de la Atención Primaria de Salud (APS) requiere políticas públicas con base en evidencias científicas. Este artículo presenta el estudio ELECT, cuyo objetivo fue identificar temas prioritarios de investigación para el fortalecimiento de la APS en el estado de São Paulo. Con la participación de especialistas y de un grupo de opinión formado por usuarios, se obtuvo una lista con los veinte principales obstáculos, así como diez temas de investigación prioritarios en la APS. Los resultados señalan problemas y temas de investigación relacionados a la organización de la gestión, la capacitación de profesionales y gestores, la valorización profesional, la creación de mecanismos de colaboración entre equipos de salud e informatización de los recursos. Se espera por lo tanto incentivar el debate en el contexto de la APS sobre el papel de la priorización de investigaciones, sus obstáculos y propuestas de investigación. Se anhela también incentivar la adopción de modelos más participativos de selección de temas de investigación.

O objetivo deste trabalho foi avaliar as respostas morfológicas do sistema radicular de quatro cultivares de café arábica submetidas a diferentes arranjos espaciais. Os espaçamentos adotados foram os de 0,40, 0,50, 0,60, 0,70 e 0,80 m entre plantas na linha, tendo-se mantido fixo 3,8 m na entrelinha. Aos 27, 35 e 39 meses após o transplantio, foram coletadas amostras de solo+raízes em três posições em relação aos caules das plantas e três profundidades. As raízes foram lavadas, coloridas, digitalizadas e processadas com o programa Safira. A cultivar Tupi RN IAC 1669-13 apresentou sistema radicular com características morfológicas relacionadas à absorção de nutrientes e de água superiores às das cultivares Catuaí Vermelho IAC 144, Catuaí Amarelo IAC 62 e Catuaí Amarelo IAC 32, para todos os arranjos espaciais testados. A redução no espaçamento aumenta a abundância de raízes por volume de solo, sem alterar a qualidade morfológica do sistema radicular, nem aprofundá-lo. Os sistemas radiculares das cultivares avaliadas são mais abundantes e apresentam superfície e comprimento específico maiores entre plantas adjacentes, seguidas das posições afastadas a 0,25 e 0,50 m do caule. A proporção de raízes na camada de 0,1 m de profundidade é maior que a na de 0,4 m, para todos os espaçamentos. A variação no espaçamento entre plantas na linha altera a morfologia do sistema radicular do café arábica.

The objective of this work was to evaluate the morphological responses of the root system of four Arabica coffee cultivars subjected to different spatial arrangements. The adopted plant spacings were 0.40, 0.50, 0.60, 0.70, and 0.80 m between plants within rows, maintaining 3.8 m constant between rows. At 27, 35, and 39 months after transplanting, samples of soil+roots were collected at three surface positions in relation to plant stems and at three depths. Roots were washed, colored, digitalized, and processed with the Safira software. The Tupi RN IAC 1669-13 cultivar showed a root system with superior morphological traits related to the absorption of nutrients and water, in comparison to those of the Catuaí Vermelho IAC 144, Catuaí Amarelo IAC 62, and Catuaí Amarelo IAC 32 cultivars, for all tested spatial arrangements. The reduction in spacing increases the amount of total root per soil volume unit, without affecting the morphological quality of the root system, nor deepening it. The root systems of the evaluated cultivars are more abundant and show higher surface and specific length between adjacent plants, followed by the 0.25 and 0.50 m positions away from the coffee stem. The proportion of roots at the 0.1 m layer is higher than at the 0.4 m layer, for all plant spacings. Changes in spacings between plants within lines affect the morphology of Arabica coffee root system.

INTRODUÇÃO: Entre 2000 e 2002, 79 amostras positivas para rotavírus foram coletadas de crianças com diarreia na Amazônia ocidental brasileira. MÉTODOS: Para a caracterização molecular dos genótipos G e P foram realizadas as reações de RT-PCR e a análise dos eletroferotipos por eletroforese em gel de poliacrilamida (PAGE). RESULTADOS: 59 amostras foram confirmadas como pertencentes ao rotavírus grupo A. Os genótipos G1P[8], G3P[8] e G4P[8] apresentaram perfis eletroforéticos longos. O genótipo G1P[8] foi encontrado em maior proporção. O eletroferotipo curto ocorreu apenas em genótipos G2. CONCLUSÕES: A proporção dos genótipos de rotavírus observada não foi diferente de outras áreas do Brasil. Este estudo é a primeira genotipagem de rotavírus na Amazônia ocidental brasileira.

INTRODUCTION: During the period from 2000 to 2002, 79 rotavirus-positive stool samples were collected from children presenting diarrhea in the Western Brazilian Amazon. METHODS: Molecular characterization of the G and P genotypes was performed using RT-PCR and electropherotyping analysis by polyacrylamide gel electrophoresis. RESULTS: A total of 59 samples were confirmed as group A rotavirus. A long electrophoretic profile was exhibited by the G1P[8], G3P[8], and G4P[8] genotypes. The G1P[8] genotype was found in greater proportion. The short electropherotype was exhibited only by G2 genotype strains. CONCLUSIONS: The proportion of the rotavirus genotypes observed was not different from that in other areas of Brazil. This study is the first genotyping of rotavirus in the Western Brazilian Amazon.

CONTEXTO E OBJETIVO: A maioria dos hipertensos em tratamento teve seu diagnóstico feito somente com medidas no consultório. O objetivo deste estudo é refazer o diagnóstico em pacientes tratados após novas medidas clínicas e monitorização ambulatorial da pressão arterial (MAPA). TIPO DE ESTUDO E LOCAL: Estudo transversal realizado em clínica de especialidade. MÉTODOS: Foram incluídos pacientes com hipertensão leve a moderada ou em tratamento anti-hipertensivo, sem lesões de órgão-alvo ou diabetes. Após suspensão do tratamento por duas a três semanas, novas medidas da pressão arterial (PA) foram feitas em duas visitas distintas. A MAPA foi feita às cegas em relação às medidas clínicas. Os limites pressóricos utilizados para o diagnóstico da hipertensão, hipertensão do avental branco, normotensão e hipertensão mascarada foram: 140 (sistólica) e 90 (diastólica) mmHg para as medidas de consultório e 135 (sistólica) e 85 (diastólica) mmHg para as médias de vigília da MAPA (MVM). RESULTADOS: Foram avaliados 101 indivíduos (70% mulheres), idade 51 ± 10 anos. PA clínica: 155 ± 18/97 ± 10 mmHg (primeira visita) e 150 ± 16/94 ± 11 mmHg (segunda visita), MVM de 137 ± 13/86 ± 10 mmHg. Sessenta e quatro (63%) pacientes foram confirmados como hipertensos, 28 (28%) como portadores de hipertensão do "avental branco", 9 (9%) como normotensos e nenhum como tendo hipertensão mascarada. Após a MAPA, 37% de presumíveis hipertensos não se enquadravam como tal. CONCLUSÃO: Este estudo revela um excesso de diagnósticos de hipertensão em hipertensos tratados. Novos procedimentos diagnósticos devem ser realizados após suspensão da medicação, com auxílio das monitorizações da PA.

CONTEXT AND OBJECTIVE: Most hypertensive subjects undergoing treatment were diagnosed solely through measurements made in the consultation office. The objective of this study was to redo the diagnosis of treated patients after new clinical measurements and ambulatory blood pressure monitoring (ABPM). DESIGN AND SETTING: Cross-sectional study conducted in an outpatient specialty clinic. METHODS: Patients with mild-to-moderate hypertension or undergoing anti-hypertensive treatment, without target organ damage or diabetes, were included. After drug withdrawal lasting 2-3 weeks, new blood pressure (BP) measurements were made during two separate visits. ABPM was performed blindly, in relation to clinical measurements. The BP thresholds used for diagnosing hypertension, white-coat hypertension, normotension and masked hypertension were: 140 (systolic) and 90 (diastolic) mmHg for office measurements and 135 (systolic) and 85 (diastolic) mmHg for mean awake ABPM (MAA). RESULTS: Evaluations were done on 101 subjects (70% women); mean age 51 ± 10 years. The clinical BP was 155 ± 18/97 ± 10 mmHg (first visit) and 150 ± 16/94 ± 11 mmHg (second visit); MAA was 137 ± 13/ 86 ± 10 mmHg. Sixty-four patients (63%) were confirmed as hypertensive, 28 (28%) as white-coat hypertensive, nine (9%) as normotensive and none as masked hypertensive. After ABPM, 37% of the presumed hypertensive patients did not fit into this category. CONCLUSION: This study showed that hypertension was overdiagnosed among hypertensive subjects undergoing treatment. New diagnostic procedures should be performed after drug withdrawal, with the aid of BP monitoring.

OBJETIVO: Avaliação dos pacientes com o quadro clínico de ceratite herpética (CH) típicas e atípicas, pela reação em cadeia da polimerase (PCR) correlacionando com o diagnóstico clínico. MÉTODOS: Foi realizada a PCR em 28 pacientes com ceratite herpética típica e atípica. RESULTADOS: A PCR foi positiva em 57,14% (n=16) do total (n=28). Nos casos de CH típica a positividade foi de 60,00% (n=12) em 20 casos. Para CH epitelial a positividade foi de 69,23% (n=9), sendo 77,78% (n=7) apenas para as lesões epiteliais dendríticas. Os casos de CH atípica apresentaram positividade de 50% (n=4) em oito casos. CONCLUSÃO: Quadro clínico típico de CH teve boa correlação com o resultado positivo observado na PCR. Entretanto, metade dos pacientes com o quadro de CH atípica apresentou PCR positivo, portanto, o exame do PCR é teste importante para o auxílio e diagnóstico da CH. No caso de CH estromal, foi demonstrado que a técnica da PCR conseguiu identificar o vírus HSV.

PURPOSE: To evaluate patients with clinically typical and atypical herpetic keratitis (HK) by means of polymerase chain reaction (PCR) as compared with the clinical diagnosis. METHODS: PCR in 28 patients with clinical symptoms of typical and atypical HK was performed. RESULTS: PCR was positive in 57.14% (n=16) of the total cases (n=28). The test was positive in 60.0% (n=12) of the 20 typical HK cases. For epithelial HK, the test was positive in 69.23% (n=9), and 77.78% (n=7) only for dendritic injuries. Atypical HK presented a positive test in 50% (n=4) of eight cases. CONCLUSION: Clinical typical picture of HK had a good correlation with the positive result of PCR, mainly for epithelial injury of the dendritic type. However, 50% of the patients with atypical HK presented positive PCR. This result showed that PCR test can provide an effective HK diagnosis. In the stromal case of HK, PCR was a useful technique to identify HSV virus.

The incidence of Shigella spp. was assessed in 877 infants from the public hospital in Rondônia (Western Amazon region, Brazil) where Shigella represents the fourth cause of diarrhea. Twenty-five isolates were identified: 18 were Shigella flexneri, three Shigella sonnei, three Shigella boydii and one Shigella dysenteriae. With the exception of S. dysenteriae, all Shigella spp. isolated from children with diarrhea acquired multiple antibiotic resistances. PCR detection of ipa virulence genes and invasion assays of bloody diarrhea and fever (colitis) were compared among 25 patients testing positive for Shigella. The ipaH and ipaBCD genes were detected in almost all isolates and, unsurprisingly, all Shigella isolates associated with colitis were able to invade HeLa cells. This work alerts for multiple antibiotic resistant Shigella in the region and characterizes presence of ipa virulence genes and invasion phenotypesin dysenteric shigellosis.

Rhoptry-associated protein 2 (RAP2) is known to be discharged from rhoptry onto the membrane surface of infected and uninfected erythrocytes (UEs) ex vivo and in vitro and this information provides new insights into the understanding of the pathology of severe anemia in falciparum malaria. In this study, a hexahistidine-tagged recombinant protein corresponding to residues 5-190 of the N-terminal of Plasmodium falciparum RAP2 (rN-RAP2) was produced using a new method of solubilization and purification. Expression was induced with D-lactose, a less expensive alternative inducer to the more common isopropyl-²-D-thio-galactopyranosidase. The recombinant protein was purified using two types of commercially-available affinity columns, iminodiacetic and nitrilotriacetic. rN-RAP2 had immunogenic potential, since it induced high titers of anti-RAP2 antibodies in mice. These antibodies recognized full-length RAP2 prepared from Triton X-100 extracts from two strains of P. falciparum. In fact, the antibody recognized a 29-kDa product of RAP2 cleavage as well as 82 and 70-kDa products of RAP1 cleavage. These results indicate that the two antigens share sequence epitopes. Our expressed protein fragment was shown to contain a functional epitope that is also present in rhoptry-derived ring surface protein 2 which attaches to the surface of both infected and UEs and erythroid precursor cells in the bone marrow of malaria patients. Serum from malaria patients who developed anemia during infection recognized rN-RAP2, suggesting that this protein fragment may be important for epidemiological studies investigating whether immune responses to RAP2 exacerbate hemolysis in falciparum malaria patients.