Specific interaction between Group B Streptococcus CC17 hypervirulent clone and phagocytes.

Streptococcus agalactiae also named Group B Streptococcus (GBS) is the most significant pathogen causing invasive infections, such as bacteremia and meningitis, in neonates. Worldwide epidemiological studies have shown that a particular clonal complex (CC) of capsular serotype III, the CC17, is strongly associated with meningitis in neonates and is therefore, designated as the hypervirulent clone. Macrophages are a permissive niche for intracellular bacteria of all GBS clones. In this study, we deciphered the specific interaction of GBS CC17 strains with macrophages. Our study revealed that CC17 strains are phagocytosed at a higher rate than GBS non-CC17 strains by human monocytes and macrophages both in cellular models and in primary cells. CC17-enhanced phagocytosis is due to an initial enhanced-attachment step to macrophages mediated by the CC17-specific surface protein HvgA and the PI-2b pilus (Spb1). We showed that two different inhibitors of scavenger receptors (fucoidan and poly(I)) specifically inhibited CC17 adhesion and phagocytosis while not affecting those of non-CC17 strains. Once phagocytosed, both CC17 and non-CC17 strains remained in a LAMP-1 positive vacuole that ultimately fuses with lysosomes where they can survive at similar rates. Finally, both strains displayed a basal egress which occurs independently from actin and microtubule networks. Our findings provide new insights into the interplay between the hypervirulent GBS CC17 and major players of the host's innate immune response. This enhanced adhesion, leading to increased phagocytosis, could reflect a peculiar capacity of the CC17 lineage to subvert the host immune defenses, establish a niche for persistence or disseminate.

Colon adenocarcinoma associated with intestinal sarcoidosis.

Sarcoidosis is a multisystem chronic inflammatory disease of unknown etiology, characterized by non-caseating granulomas. Sarcoidosis has the potential to involve every tissue in the body, which mainly affect the lymphatic system and lungs; gastrointestinal system, and particularly the colon, is an extremely rare location. We report the case of a 64-year-old male with history of pulmonary and cutaneous sarcoidosis diagnosed with neoplasm in the hepatic flexure of the colon and a polyp with high-grade dysplasia in the transverse colon by colonoscopy after a positive fecal occult blood test. The case was presented to a multidisciplinary committee and it was decided to perform a total laparoscopic colectomy and ileorectal anastomosis with histopathological evidence of infiltrating adenocarcinoma and intestinal sarcoidosis with non-caseating granulomas in the appendix, terminal ileum, colon and locoregional lymph nodes. The relationship between colon cancer and sarcoidosis is controversial, with studies showing a possible increased risk of cancer in patients with sarcoidosis, relating it to the chronic proinflammatory state of the disease. In these cases, lymph node involvement is especially important when assessing tumor extension studies, and may lead to changes in staging and, as a consequence, in the therapeutic approach.

Satisfacción e impacto del programa de especialización en Neonatología antes y durante la COVID-19 / Satisfaction and impact of the Neonatology specialization program before and during COVID-19

Fundamento: por la COVID-19, muchas facultades de Medicina implementaron nuevos planes de estudio para la formación de médicos residentes, lo cual afectó de alguna manera su proceso de formación como especialistas. Objetivo: comparar la satisfacción de los residentes de Neonatología sobre el programa de especialización y el impacto en su formación antes y durante la pandemia de COVID-19. Métodos: se realizó una investigación descriptiva con enfoque cuantitativo, en la Universidad Nacional Mayor de San Marcos en 2022. Se utilizaron métodos teóricos: análisis-síntesis, inductivo-deductivo e histórico-lógico; empírico: cuestionario a neonatólogos egresados entre 2019-2022; y matemático-estadísticos para el análisis de datos encontrados. Resultados: se analizaron 64 cuestionarios en los que se delimitaron cifras en ambos períodos: antes y durante la COVID-19, la edad promedio de los grupos de estudio fue de 38 y 37 años respectivamente; 54.8 % y 69.7 % fueron mujeres, la satisfacción fue del 100 % y 84.8 % (p=0.05), y el impacto en el efecto de sus funciones sociales fue favorable en 96.8 % y 75.8 % (p=0.03).Los tutores no promovieron un clima laboral favorable y formación profesional con ética, y durante la pandemia el impacto en su formación disminuyó de forma moderada en su desempeño y prestigio profesional. Conclusiones: la comparación efectuada evidenció muy buena satisfacción en la formación de la especialidad antes de la pandemia, no así durante la enfermedad, según la percepción de los muestreados, quienes consideraron que las actividades del año académico no se llevaron sobre la base de un plan de estudios.

Background: due to COVID-19, many medical schools implemented new study plans for the training of resident physicians, which somehow affected their training process as specialists. Objective: to compare the satisfaction of Neonatology residents about the specialization program and the impact on their training before and during the COVID-19 pandemic. Methods: a descriptive research with a quantitative approach was carried out at the Universidad Nacional Mayor de San Marcos in 2022. Theoretical methods were used: analysis-synthesis, inductive-deductive and historical-logical; empirical ones: questionnaire to neonatologists graduated from 2019 to 2022; and mathematical-statistical for the analysis of data found. Results: 64 questionnaires were analyzed in which figures were delimited in both periods: before and during COVID-19, the average age of the study groups was 38 and 37 years, respectively; 54.8% and 69.7% were women, satisfaction was 100% and 84.8% (p=0.05), and the impact on the effect of their social functions was favorable in 96.8% and 75.8% (p=0.03). Tutors did not promoted a favorable work environment and professional training with ethics, and during the pandemic the impact on their training decreased moderately on their performance and professional prestige. Conclusions: the comparison made showed very good satisfaction in the training of the specialty before the pandemic, but not during the disease, according to the perception of those sampled, who considered that the activities of the academic year were not carried out on the basis of a study plan.

Innovative and thorough practice to certify reference materials for sensory defects of olive oil.

An important problem in the olive sector is the occasional mismatch of results obtained by different tasting panels when the same olive oil sample is analysed. These discrepancies could be minimised by using reference materials (RM) for taster training. A comprehensive protocol based on the combined use of sensory and instrumental analysis for the certification of olive oil batches as RMs, developed within the framework of the project 'Operational Group INTERPANEL', is proposed. Similarity indices (R2, cosθ and NEAR) applied on GC-MS fingerprints, allow a successful homogeneity and stability assessment of produced batches. Furthermore, the use of robust statistics combined with a set of instructions developed to remove outliers were applied with excellent results on sensory data set provided by supra-panel composed by more than 100 qualified tasters. This work is the first to provide a comprehensive protocol for certification of real olive oil samples as RM for sensory analysis.

Fluorescent secreted bacterial effectors reveal active intravacuolar proliferation of Listeria monocytogenes in epithelial cells.

Real-time imaging of bacterial virulence factor dynamics is hampered by the limited number of fluorescent tools suitable for tagging secreted effectors. Here, we demonstrated that the fluorogenic reporter FAST could be used to tag secreted proteins, and we implemented it to monitor infection dynamics in epithelial cells exposed to the human pathogen Listeria monocytogenes (Lm). By tracking individual FAST-labelled vacuoles after Lm internalisation into cells, we unveiled the heterogeneity of residence time inside entry vacuoles. Although half of the bacterial population escaped within 13 minutes after entry, 12% of bacteria remained entrapped over an hour inside long term vacuoles, and sometimes much longer, regardless of the secretion of the pore-forming toxin listeriolysin O (LLO). We imaged LLO-FAST in these long-term vacuoles, and showed that LLO enabled Lm to proliferate inside these compartments, reminiscent of what had been previously observed for Spacious Listeria-containing phagosomes (SLAPs). Unexpectedly, inside epithelial SLAP-like vacuoles (eSLAPs), Lm proliferated as fast as in the host cytosol. eSLAPs thus constitute an alternative replication niche in epithelial cells that might promote the colonization of host tissues.

HIV-1 genetic diversity and resistance to antiretroviral drugs among pregnant women in Ribeirão Preto (SP), Brazil. Cross-sectional study

ABSTRACT BACKGROUND: Increasing genetic diversity of HIV-1 and emergence of drug-resistant mutations may reduce the efficacy of antiretroviral therapy and prophylaxis that are used to prevent mother-to-child transmission. The aim of this study was to assess the genetic diversity and prevalence of drug-resistant mutations among HIV-infected pregnant women. DESIGN AND SETTING: Cross-sectional study at an outpatient clinic for infectious diseases within gynecology and obstetrics. METHODS: This study evaluated the dynamics of HIV-1 subtypes and the prevalence of transmitted and acquired drug-resistant mutations among 38 HIV-infected pregnant women (20 previously exposed to antiretroviral therapy and 18 naive), in Ribeirão Preto (SP), Brazil, between 2010 and 2011. Genotyping was performed by means of molecular sequencing of the protease and reverse transcriptase regions of the HIV-1 pol gene. RESULTS: Subtype B was identified in 84.2% of the samples, recombinant forms between B and F in 7.9%, subtype F1 in 5.3% and the recombinant form K/F in 2.6%. No mutation associated with transmitted drug resistance was detected in the samples from the naive pregnant women, whereas mutations associated with acquired drug resistance were found in 35.0% of the pregnant women previously exposed to antiretroviral therapy. CONCLUSION: The results showed that subtype B predominated, while there was low prevalence of sequences with transmitted drug resistance.

Single Nucleotide Polymorphisms in Cellular Drug Transporters Are Associated with Intolerance to Antiretroviral Therapy in Brazilian HIV-1 Positive Individuals.

Adverse reactions are the main cause of treatment discontinuation among HIV+ individuals. Genes related to drug absorption, distribution, metabolism and excretion (ADME) influence drug bioavailability and treatment response. We have investigated the association between single nucleotide polymorphisms (SNPs) in 29 ADME genes and intolerance to therapy in a case-control study including 764 individuals. Results showed that 15 SNPs were associated with intolerance to nucleoside and 11 to non-nucleoside reverse transcriptase inhibitors (NRTIs and NNRTIs), and 8 to protease inhibitors (PIs) containing regimens under alpha = 0.05. After Bonferroni adjustment, two associations remained statistically significant. SNP rs2712816, at SLCO2B1 was associated to intolerance to NRTIs (ORGA/AA = 2.37; p = 0.0001), while rs4148396, at ABCC2, conferred risk of intolerance to PIs containing regimens (ORCT/TT = 2.64; p = 0.00009). Accordingly, haplotypes carrying rs2712816A and rs4148396T alleles were also associated to risk of intolerance to NRTIs and PIs, respectively. Our data reinforce the role of drug transporters in response to HIV therapy and may contribute to a future development of personalized therapies.

Case 224: Cardiac Involvement in Erdheim-Chester Disease.

History A 61-year-old man with no relevant medical history was admitted to the emergency department with symptoms of congestive heart failure and a 1-week history of chest pain, progressive dyspnea, abdominal swelling, bipedal edema, and anorexia. Laboratory test results, including complete blood count and electrolyte, creatinine, creatine phosphokinase, and troponin T levels were normal. Electrocardiographic findings were unremarkable. Initial chest radiography showed an enlarged heart with bilateral pleural effusion. Transthoracic echocardiography revealed an irregular right atrial mass and moderate to severe pericardial effusion. The patient subsequently underwent computed tomography (CT) of the chest, abdomen, and pelvis followed by cardiac magnetic resonance (MR) imaging for further evaluation of the atrial mass. Because of the suspected diagnosis, conventional radiography of the skeleton was performed.

Cellular distribution and subcellular localization of spatacsin and spastizin, two proteins involved in hereditary spastic paraplegia.

Truncating mutations in the SPG11 and SPG15 genes cause complicated spastic paraplegia, severe neurological conditions due to loss of the functions of spatacsin and spastizin, respectively. We developed specific polyclonal anti-spatacsin (SPG11) and anti-spastizin (SPG15) antisera, which we then used to explore the intracellular and tissue localizations of these proteins. We observed expression of both proteins in human and rat central nervous system, which was particularly strong in cortical and spinal motor neurons as well as in retina. Both proteins were also expressed ubiquitously and strongly in embryos. In cultured cells, these two proteins had similar diffuse punctate, cytoplasmic and sometimes nuclear (spastizin) distributions. They partially co-localized with multiple organelles, particularly with protein-trafficking vesicles, endoplasmic reticulum and microtubules. Spastizin was also found at the mitochondria surface. This first study of the endogenous expression of spatacsin and spastizin shows similarities in their expression patterns that could account for their overlapping clinical phenotypes and involvement in a common protein complex.

Prevalence of HIV type 1 drug resistance mutations in treatment-naïve and experienced patients from resource-limited settings with universal access to antiretroviral therapy: a survey in two small Brazilian cities

Concerns have been raised that universal availability of antiretroviral agents in resource-limited settings might lead to the emergence and spread of resistant strains. We present the largest survey on human immunodeficiency virus type 1 (HIV-1) resistance among treatment-naïve and experienced patients followed in small, relatively underprivileged cities in Brazil with universal availability to standard of care antiretroviral combinations. Samples were collected between 2004 and 2006 from 95 patients followed in the cities of Saquarema and Santo Antonio de Pádua, state of Rio de Janeiro. A proviral fragment encompassing protease and reverse transcriptase (RT) regions was generated and drug susceptibility level was inferred. Among 50 strains from drug-naïve subjects, one (2 percent) had intermediate-level resistance to RT inhibitors. Among 38 patients on therapy as of sampling, 28 (73.7 percent) had plasma viral load (PVL) below detection limit (26 of whom without evidence of resistance mutations) and 11 (28.9 percent) harbored strains with reduced susceptibility. Only two strains harbored both protease and RT inhibitor mutations. Among seven patients who were off-treatment as of sampling, two (28.5 percent) harbored strains with reduced susceptibility to RT inhibitors. The relatively high frequency of undetectable PVL among patients on treatment and the overall low prevalence of resistance-associated mutations are reassuring. Continued surveillance, however, is necessary.

Human immunodeficiency virus type- 1 subtypes of infected patients in Espírito Santo, Brazil

Genetic variability of human immunodeficiency virus type - 1(HIV-1) is a potential threat for both diagnosis and treatment of HIV/AIDS, as well as the development of effective vaccines. Up to now, HIV subtypes circulating among HIV-positive patients in the state of Espírito Santo were not known. In the present study, blood samples from 100 therapy-naïve HIV-1 infected patients were collected and the HIV subtype was determined through the Heteroduplex Mobility Assay (HMA). Ninety-seven out of 100 studied samples were subtyped by HMA, 73 samples (75.2 percent) were from subtype B, 9 (9.3 percent) from subtype F, 3 (3.1 percent) from subtype C, 6 (6.2 percent) Benv/Fgag, and another 6 (6.2 percent) Fenv/Bgag, what suggests that recombinant viruses were present in the studied samples. Twenty-eight percent of the subtype B samples were represented by the Brazilian B" subtype, which were identified by RFLP with Fok I. Data presented here demonstrate that the epidemiological characteristics of the HIV epidemic in the state of Espírito Santo are similar to those from the other Southeastern states and helped to better understand the genetic polymorphism of HIV in Brazil.

Brote epidémico de fascioliasis invasiva: estudio de una familia / Epidemic outbreak of invasive fasciolasis: study of a family

Se estudiaron 6 casos de una familia procedente del área rural, con antecedentes de ingestión de berro silvestre, para exponer nuestras observaciones clínicas y terapéuticas sobre esta infección. Se observó que unas semanas más tarde a la ingestión se presentaron síntomas de astenia, dolor en hipocondrio derecho, fiebre, hepatomegalia. Se realizó leucograma, recuento global de eosinófilos, aminotransferasas, fosfatasa alcalina, antígeno de superficie, anti HVC, serodiagnóstico para fasciola (ELISA indirecto), heces fecales seriadas, drenaje biliar, laparoscopia y biopsia hepática y colangiopancreatografía transduodenal. Se estudió solamente mediante exámenes de laboratorio el caso de un niño. Se halló un recuento global de eosinófilos elevado en todos los casos. Se detectaron huevos de fasciola en 2 casos: uno en heces fecales y otro, en bilis. Se demostró, mediante laparoscopia, la presencia de lesiones de larva migrans visceral en la superficie hepática, que se comportaron histológicamente como granulomas. La colangiopancreatografía transduodenal resultó un método útil en el diagnóstico y tratamiento de algunos casos. Se indicó tratamiento con emetina y prazicuantel indistintamente

Contribuição ao estudo da diversidade do HIV-1 no Brasil: caracterização genotípica e fenotípica de amostras de HIV-1 obtidas em diferentes regiões geográficas / Contribution to the study of HIV-1 diversity in Brazil: genotypic and phenotypic characterization of HIV-1 samples obtained from different geographical regions

Desde o estabelecimento em cultura do primeiro isolado de HIV-l no Brasil (Galvão-Castro e cols., 1987), já podíamos perceber um polimorfismo genético e biológico em amostras obtidas no início da epidemia de AIDS no país, indicando que esta heterogeneidade seria similar a descrita para o HIV-l em outras partes do mundo (Couto-Fernandez e cols., 1992). Nos anos subsequentes, buscando um melhor detalhamento da estrutura genética destes isolados, entre outros, sequenciamos um fragmento genômico de aproximadamente 860pb correspondentes as regiões V3, V4, V5 e parte da gp41 do envelope viral de 6 amostras coletadas entre 1987 e 1989 (documento 1). A análise filo genética mostrou que estas amostras brasileiras eram do subtipo B, com níveis de diversidade variando entre 5,9 e 13, 1%. A análise das seqüências de aminoácidos da alça V3 revelou que 3 amostras possuíam o motivo GPGR no topo da alça, característico de isolados Norte Americanos/Europeus, enquanto que nas outras três existia uma substituição da prolina pelo triptofano (W), metionina (M) e fenilalalina (F), respectivamente. O grau de diversidade antigênica, avaliado pela reatividade sorológica de 114 soros obtidos de pacientes do Rio de Janeiro, coletados no período de 1990 a 1992, frente à diferentes peptídeos sintéticos subtipo-específicos, mostrou que 60,5% deles reagiram com peptídeos do subtipo B e 15,8% mostraram reatividade específica para o peptídeo Bra-cons (GWG). Um elevado percentual de soros (37,7%) não mostrou reatividade com os diferentes peptídeos usados. Buscando conhecer o espectro da diversidade do HIV-1 nos sítios selecionados para futuros testes de vacinas anti-HIV/AIDS, foi estabelecida a Rede Nacional de Laboratórios para Isolamento e Caracterização do HIV-1 no Brasil que, permitiu uma análise abrangente e sistematizada de amostras coletadas em diferentes regiões do país. Estes estudos, realizados primeiramente em amostras do Rio de Janeiro (documento 2) e, subseqüentemente,...

Seroprevalence of HIV, HTLV-I/II and other perinatally-transmitted pathogens in Salvador, Bahia

A obtencao de dados epidemiologicos e de fundamental importancia para o estabelecimento de politicas em Saude Pubica. No Brasil, essas informacoes sao escassas, principalmente na regiao Nordeste. Para se obter alguns desses dados, avaliamos a soroprevalencia de algumas infeccoes de transmissao perinatal, em cerca de 1024 gestantes de baixa renda, em Salvador, Bahia...

Molecular and biological diversity of HIV-1 in Brazil

To determine the genomic polymorphism and biological properties present in HIV-1 Brazilian isolates, were analyzed five viral isolates obtained from patients residing in Rio de Janeiro (P1 and P5), Säo Paulo (P3) and Bahia (P2 and P4) states. For each viral isolate in vitro characteristics such as replication rate, syncytium-inducing capacity and cell death were observed in lymphoblastoid (H9, CEM and peripheral blood mononuclear cells) as well as monocytoid (U937) cells. In addition, the evaluation of the restriction fragment lenght polymorphism of these isolates was also performed using a panel of endonucleases such as Hind III, Bgl II, Sac I, Pst I, Kpn I and Eco RI. One of the isolates (P1), showed the highest phenotypic and genotypic divergence, when compared to others. The results found suggest a HIV heterogeneity in Brazil similar to that already described in other regions of the world