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.

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.

Attending pediatric acutely ill patients at home: families' socioeconomic characterization, expectations, and experiences.

BACKGROUND: SJD a Casa is an acute pediatric hospital-at-home program that was initiated in 2019. For a thorough understanding of acute pediatric homecare programs, an analysis of all related factors, including the medical, social, and economic aspects as well as the family's experience, is essential. However, no previous study has attempted a comprehensive evaluation of this topic in relation to a complex program such as ours (in terms of the diseases and treatments offered). In this study, we aimed to finely characterize the population that opts for pediatric homecare programs and obtain a thorough understanding of the families' needs, which will improve our understanding of the program and potentially reveal possible deficiencies. METHODS: This prospective quantitative and qualitative study involved collection of ordinal data as well as statements made by the caregivers of patients undergoing homecare. A total of 372/532 families were asked to answer two independent questionnaires (preadmission and postadmission) that evaluated their socioeconomic characteristics; expectations and experiences; and factors influencing the preference for homecare. The results were presented as frequencies and comparisons (Fisher's exact test). RESULTS: The families had an adequate social network and a less-than-expected workload, and most families responded that they would have repeated the experience despite the workload. The expectations regarding the caregiver's well-being at home were better than the actual situation, since some caregivers experienced anxiety or fear. The rating for homecare was better than that for the inpatient care offered before the homecare transfer. CONCLUSIONS: Families included in the program were content with the homecare program and mostly responded that they would repeat the experience if needed. Although the duration of the program was short-term, some caregivers may experience symptoms of burnout like anxiety, which should be taken into consideration. Despite its limitations, this study offers the possibility of improving our service portfolio by focusing on vulnerable families' access to the program and the caregiver's risk of burnout.

Moderate prevalence of HIV-1 transmitted drug resistance mutations in southern Brazil.

BACKGROUND: Despite the advances in therapy, the occurrence of drug-resistant human immunodeficiency virus type 1 (HIV-1) is a major obstacle to successful treatment. This study aimed to characterize the genetic diversity and to determine the prevalence of transmitted drug resistance mutations (TDRM) between individuals recently or chronically diagnosed with HIV-1 from Paraná, Brazil. METHODS: A total of 260 HIV-1 positive antiretroviral therapy-naïve patients were recruited to participate on the study, of which 39 were recently diagnosed. HIV-1 genotyping was performed using sequencing reaction followed by phylogenetic analyses to determine the HIV-1 subtype. TDRM were defined using the Calibrated Population Resistance Tool program. RESULTS: The HIV-1 subtypes frequency found in the studied population were 54.0% of subtype B, 26.7% subtype C, 6.7% subtype F1 and 12.7% recombinant forms. The overall prevalence of TDRM was 6.7%, including 13.3% for recently diagnosed subjects and 5.9% for the chronic group. CONCLUSIONS: The prevalence of resistance mutations found in this study is considered moderate, thus to perform genotyping tests before the initiation of antiretroviral therapy may be important to define the first line therapy and contribute for the improvement of regional prevention strategies for epidemic control.

Treatment of a chronic anal fissure with a botulin toxin A injection and fissurectomy.

INTRODUCTION: pharmacological treatment of a chronic anal fissure (CAF) achieves healing in half of cases and lateral internal sphincterotomy (LIS) is the definite treatment. The objective of this study was to assess the combination of fissurectomy and botulin toxin A (BTA) injection. METHODS: this was a retrospective study of 54 patients with anal sphincter hypertonia and CAF treated with an injection of BAT and fissurectomy, after an unsuccessful management with topical nitroglycerin (NGT) for eight weeks. Fissurectomy and an injection of BTA (33 or 50 units) in the internal anal sphincter was performed during the same session. The main outcome measure was the healing rate, with incontinence and the need of LIS as secondary outcomes. RESULTS: two patients were excluded from the study, one due to Crohn's disease and the other was lost to follow-up. Of the 52 patients included in the study, there were 36 females (70%) and 16 (30%) males, with a mean age of 49 years (range 22-75). Fissure healing was initially achieved in 49 patients (94.2%) and LIS was required in the remaining three patients (5.8%). After initial healing, 18 patients (34.7%) developed 23 recurrences at a mean time of 27 months (5-83 months). Of these patients, healing with conservative sphincter measures was obtained in eleven cases (NGT in eight and repeat fissurectomy and BAT in three); two patients are currently under treatment with NGT and five underwent LIS. CONCLUSIONS: BTA injection associated with fissurectomy is a safe and effective procedure in patients with CAF, avoiding the need of LIS in a high percentage of patients.

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.

Multilocus sequence analysis of the genus Citrobacter and description of Citrobacter pasteurii sp. nov.

Strains originating from various sources and classified as members of the genus Citrobacter within the family Enterobacteriaceae were characterized by sequencing internal portions of genes rpoB, fusA, pyrG and leuS, 16S rRNA gene sequencing, average nucleotide identity (ANI) of genomic sequences and biochemical tests. Phylogenetic analysis based on the four housekeeping genes showed that the 11 species of the genus Citrobacter with validly published names are well demarcated. Strains CIP 55.13(T) and CIP 55.9 formed a distinct branch associated with Citrobacter youngae. The ANI between CIP 55.9 and CIP 55.13(T) was 99.19%, whereas it was 94.75% between CIP 55.13(T) and strain CIP 105016(T) of the species C. youngae, the most closely related species. Biochemical characteristics consolidated the fact that the two isolates represent a separate species, for which the name Citrobacter pasteurii sp. nov. is proposed. The type strain is CIP 55.13(T) ( =DSM 28879(T) =Na 1a(T)).

Paenibacillus faecis sp. nov., isolated from human faeces.

A spore-forming, rod-shaped Gram-strain-positive bacterium, strain 656.84T, was isolated from human faeces in 1984. It contained anteiso-C15 : 0 as the major cellular fatty acid, meso-diaminopimelic acid was found in the cell wall peptidoglycan, the polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and aminophospholipids as the major components, and the predominant menaquinone was MK-7. The DNA G+C content was 52.9 mol%. The results of comparative 16S rRNA gene sequence studies placed strain 656.84T within the genus Paenibacillus. Its closest phylogenetic relatives were Paenibacillus barengoltzii and Paenibacillus timonensis. Levels of DNA-DNA relatedness between strain 656.84T and Paenibacillus timonensis CIP 108005T and Paenibacillus barengoltzii CIP 109354T were 17.3 % and 36.8 %, respectively, indicating that strain 656.84T represents a distinct species. On the basis of phenotypic and genotypic results, strain 656.84T is considered to represent a novel species within the genus Paenibacillus, for which the name Paenibacillus faecis sp. nov. is proposed; the type strain is 656.84T ( = DSM 23593T = CIP 101062T).

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.

HIV-1 Diversity and Drug Resistance in Treatment-Naïve Children and Adolescents from Rio de Janeiro, Brazil.

The human immunodeficiency virus type 1 (HIV-1) can be transmitted via parenteral, sexual, or vertical exposure routes. The number of HIV-1 cases detected yearly in children and adolescents in Brazil did not decrease over the last decade, representing ~5% of total cases described in the country. In recent years, the HIV-1 diversity and the prevalence of transmitted drug resistance mutations (TDRM) are moving toward a marked increase. In this study, we retrospectively evaluated the diversity of HIV-1 subtypes and the TDRM prevalence in 135 treatment-naïve HIV-1 vertically infected children and adolescents born in between 1993 and 2012. These children were assessed in either 2001-2007 or 2008-2012 when they were 0 to 17 years old. The individuals assessed in 2001-2007 (n = 38) had median CD4+ T cell counts of 1218 cells/mm3 (IQR: 738-2.084) and median HIV-1 plasma viral load of 4.18 log10 copies/mL (IQR: 3.88-4.08). The individuals (n = 97) evaluated in 2008-2012 showed median CD4+ T cell counts of 898.5 cells/mm3 (IQR: 591.3-1.821) and median HIV-1 plasma viral load of 4.69 log10 copies/mL (IQR: 4.26-5.33). A steady decrease in the median CD4 T+ cell counts was observed with age progression, as expected. The majority HIV-1 pol sequences (87%) were classified as pure HIV-1 subtypes (77% subtype B, 9% subtype F1 and 1.5% subtype C), while 13% of sequences were classified as recombinants (CRF45_cpx, n = 4; CRF28/29_BF1, n = 2; CRF02_AG, n = 1; CRF40_BF1, n = 1, CRF99_BF1, n = 1, URF_BF1, n = 8). The overall prevalence of TDRM was 14% (19/135), conferring resistance to the nucleoside reverse transcriptase inhibitors (NRTI, 13/135-9.6%), non-nucleoside reverse transcriptase inhibitors (NNRTI, 8/135-5.9%), and protease inhibitors (PI, 2/135-1.5%). The main TDRM observed for NNRTI was the K103N (n = 8), while the mutations T215I/Y/D/E (n = 7) and M184V (n = 4) were the main TDRM for NRTI. Only two TDRM were observed for PI in one individual each (M46I and V82A). Most TDRM were found in the HIV-1 subtype B (84%) sequences. This study reveals an HIV-1 epidemic with high diversity and moderate prevalence of TDRM in the pediatric population of Rio de Janeiro, indicating the existence of possible problems in the clinical management of prophylactic therapy to prevent mother-to-child transmission and future treatment options for the affected children.

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.

HIV-1 molecular diversity in Brazil unveiled by 10 years of sampling by the national genotyping network.

HIV-1 has diversified into several subtypes and recombinant forms that are heterogeneously spread around the world. Understanding the distribution of viral variants and their temporal dynamics can help to design vaccines and monitor changes in viral transmission patterns. Brazil has one of the largest HIV-1 epidemics in the western-world and the molecular features of the virus circulating in the country are still not completely known. Over 50,000 partial HIV-1 genomes sampled between 2008 and 2017 by the Brazilian genotyping network (RENAGENO) were analyzed. Sequences were filtered by quality, duplicate sequences per patient were removed and subtyping was performed with online tools and molecular phylogeny. Association between patients' demographic data and subtypes were performed by calculating the relative risk in a multinomial analysis and trends in subtype prevalence were tested by Pearson correlation. HIV-1B was found to be the most prevalent subtype throughout the country except in the south, where HIV-1C prevails. An increasing trend in the proportion of HIV-1C and F1 was observed in several regions of the country, while HIV-1B tended to decrease. Men and highly educated individuals were more frequently infected by HIV-1B and non-B variants were more prevalent among women with lower education. Our results suggest that socio-demographic factors partially segregate HIV-1 diversity in Brazil while shaping viral transmission networks. Historical events could explain a preferential circulation of HIV-1B among men who have sex with men (MSM) and non-B variants among heterosexual individuals. In view of an increasing male/female ratio of AIDS cases in Brazil in the last 10-15 years, the decrease of HIV-1B prevalence is surprising and suggests a greater penetrance of non-B subtypes in MSM transmission chains.

Long-lasting severe immune dysfunction in Ebola virus disease survivors.

Long-term follow up studies from Ebola virus disease (EVD) survivors (EBOV_S) are lacking. Here, we evaluate immune and gene expression profiles in 35 Guinean EBOV_S from the last West African outbreak, a median of 23 months (IQR [18-25]) after discharge from treatment center. Compared with healthy donors, EBOV_S exhibit increases of blood markers of inflammation, intestinal tissue damage, T cell and B cell activation and a depletion of circulating dendritic cells. All survivors have EBOV-specific IgG antibodies and robust and polyfunctional EBOV-specific memory T-cell responses. Deep sequencing of the genes expressed in blood reveals an enrichment in 'inflammation' and 'antiviral' pathways. Integrated analyses identify specific immune markers associated with the persistence of clinical symptoms. This study identifies a set of biological and genetic markers that could be used to define a signature of "chronic Ebola virus disease (CEVD)".

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.