Molecular epidemiology and antimicrobial resistance in Clostridioides difficile strains isolated from children and adolescents in a tertiary referral pediatric hospital in Fortaleza, Brazil.

BACKGROUND: C. difficile has been increasingly reported as a cause of gastrointestinal disease in children, ranging from mild self-limiting diarrhea to severe conditions such as pseudomembranous colitis and toxic megacolon. Only two pediatric research groups reported the presence of C. difficile infection in Brazilian children, but no previous research has examined C. difficile infection among children in northeastern Brazil. This prospective cross-sectional study investigated the molecular epidemiology and antimicrobial resistance of C. difficile strains isolated from children and adolescents with diarrhea referred to a tertiary pediatric hospital in Brazil while exploring the associated risk factors. RESULTS: Toxin positivity or C. difficile isolation was found in 30.4 % (17/56) samples. C. difficile was isolated from 35 % (6/17) samples. Four toxigenic strains were identified (tpi+, tcdA+, tcdB+, cdtB-, without tcdC deletions) belonging to PCR ribotypes and PFGE-pulsotypes: 046 (new pulsotype 1174), 106 (NAP11), 002 (new pulsotype 1274), 012 (new pulsotype NML-1235). Two of the six isolates belonging to ribotypes 143 and 133 were non-toxigenic. All toxigenic strains were sensitive to metronidazole and vancomycin. Regarding the clinical manifestation, diarrhea lasted an average of 11 days, ranging from 3 to 50 days and was often associated with mucus and/or blood. All six patients from whom the C. difficile was isolated had a chronic disease diagnosis, with these comorbidities as the main risk factors. CONCLUSION: Our study enhances our understanding of the present epidemiological landscape of C. difficile-associated diarrhea (CDI) among children in northeastern Brazil, reveling a substantial CDI frequency of 30.4 %, with toxigenic strains detected in 76.4 % of cases, highlighting a higher prevalence compared to earlier Brazilian studies. In the globalized world, an understanding of disease-generating strains, the associated risk factors, clinical manifestation, and antimicrobial sensitivity has fundamental epidemiological importance and draws attention to preventive measures, allowing for more decisive action.

First Transcriptome Analysis of Hepatoblastoma in Brazil: Unraveling the Pivotal Role of Noncoding RNAs and Metabolic Pathways.

Hepatoblastoma stands as the most prevalent liver cancer in the pediatric population. Characterized by a low mutational burden, chromosomal and epigenetic alterations are key drivers of its tumorigenesis. Transcriptome analysis is a powerful tool for unraveling the molecular intricacies of hepatoblastoma, shedding light on the effects of genetic and epigenetic changes on gene expression. In this study conducted in Brazilian patients, an in-depth whole transcriptome analysis was performed on 14 primary hepatoblastomas, compared to control liver tissues. The analysis unveiled 1,492 differentially expressed genes (1,031 upregulated and 461 downregulated), including 920 protein-coding genes (62%). Upregulated biological processes were linked to cell differentiation, signaling, morphogenesis, and development, involving known hepatoblastoma-associated genes (DLK1, MEG3, HDAC2, TET1, HMGA2, DKK1, DKK4), alongside with novel findings (GYNG4, CDH3, and TNFRSF19). Downregulated processes predominantly centered around oxidation and metabolism, affecting amines, nicotinamides, and lipids, featuring novel discoveries like the repression of SYT7, TTC36, THRSP, CCND1, GCK and CAMK2B. Two genes, which displayed a concordant pattern of DNA methylation alteration in their promoter regions and dysregulation in the transcriptome, were further validated by RT-qPCR: the upregulated TNFRSF19, a key gene in the embryonic development, and the repressed THRSP, connected to lipid metabolism. Furthermore, based on protein-protein interaction analysis, we identified genes holding central positions in the network, such as HDAC2, CCND1, GCK, and CAMK2B, among others, that emerged as prime candidates warranting functional validation in future studies. Notably, a significant dysregulation of non-coding RNAs (ncRNAs), predominantly upregulated transcripts, was observed, with 42% of the top 50 highly expressed genes being ncRNAs. An integrative miRNA-mRNA analysis revealed crucial biological processes associated with metabolism, oxidation reactions of lipids and carbohydrates, and methylation-dependent chromatin silencing. In particular, four upregulated miRNAs (miR-186, miR-214, miR-377, and miR-494) played a pivotal role in the network, potentially targeting multiple protein-coding transcripts, including CCND1 and CAMK2B. In summary, our transcriptome analysis highlighted disrupted embryonic development as well as metabolic pathways, particularly those involving lipids, emphasizing the emerging role of ncRNAs as epigenetic regulators in hepatoblastomas. These findings provide insights into the complexity of the hepatoblastoma transcriptome and identify potential targets for future therapeutic interventions.

Molecular epidemiology and antimicrobial resistance in Clostridioides difficile strains isolated from children and adolescents in a tertiary referral pediatric hospital in Fortaleza, Brazil

Abstract Background C. difficile has been increasingly reported as a cause of gastrointestinal disease in children, ranging from mild self-limiting diarrhea to severe conditions such as pseudomembranous colitis and toxic megacolon. Only two pediatric research groups reported the presence of C. difficile infection in Brazilian children, but no previous research has examined C. difficile infection among children in northeastern Brazil. This prospective cross-sectional study investigated the molecular epidemiology and antimicrobial resistance of C. difficile strains isolated from children and adolescents with diarrhea referred to a tertiary pediatric hospital in Brazil while exploring the associated risk factors. Results Toxin positivity or C. difficile isolation was found in 30.4 % (17/56) samples. C. difficile was isolated from 35 % (6/17) samples. Four toxigenic strains were identified (tpi+, tcdA+, tcdB+, cdtB-, without tcdC deletions) belonging to PCR ribotypes and PFGE-pulsotypes: 046 (new pulsotype 1174), 106 (NAP11), 002 (new pulsotype 1274), 012 (new pulsotype NML-1235). Two of the six isolates belonging to ribotypes 143 and 133 were non-toxigenic. All toxigenic strains were sensitive to metronidazole and vancomycin. Regarding the clinical manifestation, diarrhea lasted an average of 11 days, ranging from 3 to 50 days and was often associated with mucus and/or blood. All six patients from whom the C. difficile was isolated had a chronic disease diagnosis, with these comorbidities as the main risk factors. Conclusion Our study enhances our understanding of the present epidemiological landscape of C. difficile-associated diarrhea (CDI) among children in northeastern Brazil, reveling a substantial CDI frequency of 30.4 %, with toxigenic strains detected in 76.4 % of cases, highlighting a higher prevalence compared to earlier Brazilian studies. In the globalized world, an understanding of disease-generating strains, the associated risk factors, clinical manifestation, and antimicrobial sensitivity has fundamental epidemiological importance and draws attention to preventive measures, allowing for more decisive action.

Insulin in combination with pioglitazone prevents advanced cachexia in 256-Walker tumor-bearing rats: effect is greater than treatment alone and is associated with improved insulin sensitivity.

BACKGROUND: Insulin (INS) resistance and hypoinsulinemia commonly observed in cancer-carrying, can contribute to cachexia. However, the effects of INS and INS sensitizers, such as pioglitazone (PIO), particularly when used in combination therapy, on cancer cachexia have not been evaluated sufficiently. We investigated the effects of INS and PIO, at various doses, either isolated or combined, on cachexia in Walker-256 tumor-bearing rats (TB rats). METHODS: INS or INS + PIO were administered in TB rats, for 6 or 12 days, starting from the day of tumor cells inoculation. RESULTS: INS at 18 or 27 U/kg (12-days treatment), but not 9 U/kg, reduced fat loss and slightly prevented weight loss. However, INS 18 U/kg + PIO 5, 10, 20, or 40 mg/kg (6 or 12-day treatment) reduced fat loss and markedly prevented weight loss but did not affect muscle wasting. While TB rats lost weight (37.9% in 12 days), TB rats treated with INS 18 U/kg + PIO 5 mg/kg showed pronounced weight gain (73.7%), which was greater than the sum (synergism) of the weight gains promoted by isolated treatments with INS 18 U/kg (14.7%) or PIO 5 mg/kg (13.1%). The beneficial effect of the INS 18 U/kg + PIO 5 mg/kg on weight loss was associated with improved INS sensitivity, as indicated by the higher blood glucose clearance constant (kITT), decreased levels of free fatty acids and triacylglycerols (INS resistance-inducing factors) in the blood, and increased expression of p-Akt (INS signaling pathway protein) in adipose tissue. CONCLUSIONS: The combined treatment with INS 18 U/kg + PIO 5 mg/kg was more effective in preventing advanced cachexia in TB rats than each treatment alone, emerging as the best approach, considering the lower dosage and higher efficacy. This combination completely preserved adipose mass and markedly reduced weight loss through a synergistic mechanism linked to improved insulin sensitivity. These findings provide new insights into the importance of drug combinations in effectively combating fat loss in advanced cachexia.

Prevalence and clinical implications of germline pathogenic variants in cancer predisposing genes in young patients across sarcoma subtypes.

BACKGROUND: Sarcomas are a rare and diverse group of cancers occurring mainly in young individuals for which an underlying germline genetic cause remains unclear in most cases. METHODS: Germline DNA from 177 children, adolescents and young adults with soft tissue or bone sarcomas was tested using multigene panels with 113 or 126 cancer predisposing genes (CPGs) to describe the prevalence of germline pathogenic/likely pathogenic variants (GPVs). Subsequent testing of a subset of tumours for loss of heterozygosity (LOH) evaluation was performed to investigate the clinical and molecular significance of these variants. RESULTS: GPVs were detected in 21.5% (38/177) of the patients (15.8% in children and 21.6% in adolescents and young adults), with dominant CPGs being altered in 15.2% overall. These variants were found in genes previously associated with the risk of developing sarcomas (TP53, RB1, NF1, EXT1/2) but also in genes where that risk is still emerging/limited (ERCC2, TSC2 and BRCA2) or unknown (PALB2, RAD50, FANCM and others). The detection rates of GPVs varied from 0% to 33% across sarcoma subtypes and GPV carriers were more likely to present more than one primary tumour than non-carriers (21.1%×6.5%; p=0.012). Loss of the wild-type allele was detected in 48% of tumours from GPV carriers, mostly in genes definitively associated with sarcoma risk. CONCLUSION: Our findings reveal that a high proportion of young patients with sarcomas presented a GPV in a CPG, underscoring the urgency of establishing appropriate genetic screening strategies for these individuals and their families.

Effect of promethazine on biofilms of gram-positive cocci associated with infectious endocarditis.

This study evaluated the antimicrobial activity of promethazine against Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus mutans and its effect on the antimicrobial susceptibility of biofilms grown in vitro and ex vivo on porcine heart valves. Promethazine was evaluated alone and in combination with vancomycin and oxacillin against Staphylococcus spp. and vancomycin and ceftriaxone against S. mutans in planktonic form and biofilms grown in vitro and ex vivo. Promethazine minimum inhibitory concentration range was 24.4-95.31 µg/mL and minimum biofilm eradication concentration range was 781.25-3.125 µg/mL. Promethazine interacted synergistically with vancomycin, oxacillin and ceftriaxone against biofilms in vitro. Promethazine alone reduced (p < 0.05) the CFU-counts of biofilms grown on heart valves for Staphylococcus spp., but not for S. mutans, and increased (p < 0.05) the activity of vancomycin, oxacillin and ceftriaxone against biofilms of Gram-positive cocci grown ex vivo. These findings bring perspectives for repurposing promethazine as adjuvant in the treatment of infective endocarditis.

O fortalecimento do patriarcado e a desvalorização do trabalho de cuidado no capitalismo / The strengthenin of patriarchy and the devaluation of care work in capitalism

Resumo: Este artigo busca compreender como os processos de colonização europeia e de acumulação primitiva do capital estão relacionados à destinação de atividades de cuidado para as mulheres, e a simultânea invisibilidade e desvalorização dessas experiências femininas. À luz de autoras feministas, são discutidos como a expansão do capitalismo e a colonização desencadearam o fortalecimento do patriarcado e a potencialização da divisão sexual do trabalho.

Abstract: This article seeks to understand how the processes of European colonization and primitive capital accumulation are related to the allocation of care activities for women and the simultaneous invisibility and devaluation of these female experiences. In the light of feminist authors, it is discussed how the expansion of capitalism and colonization triggered the strengthening of patriarchy and the potentiation of the sexual division of labor.

Perfil Socioeconômico de Pessoas com Câncer de Laringe e Cavidade Oral em Tratamento no Instituto Nacional de Câncer / Socioeconomic Profile of People with Cancer of the Larynx and Oral Cavity Being Treated at National Cancer Institute / Perfil Socioeconómico de las Personas con Cáncer de Laringe y Cavidad Oral en Tratamiento en el Instituto Nacional de Cáncer

Introdução: O câncer é um dos principais problemas de saúde pública do mundo. O câncer de cabeça e pescoço apresenta como principais fatores de risco o tabagismo, o etilismo, entre outros fatores associados ao modelo econômico vigente, explicitando a associação entre desigualdades socioeconômicas e incidência/mortalidade. Objetivo: Conhecer o perfil socioeconômico das pessoas com câncer de laringe e cavidade oral, matriculadas na clínica de cabeça e pescoço do HCI/INCA no período de fevereiro a julho de 2017. Método: O caminho metodológico empregado se divide em dois eixos: revisão de literatura narrativa e definição conceitual de terminologias utilizadas na pesquisa; e questionário aplicado com pessoas adoecidas. Trata-se de uma pesquisa exploratória que visa levantar elementos qualitativos para a construção do perfil socioeconômico. Resultados: Os resultados apresentam o perfil socioeconômico, demonstram a magnitude das condições socioeconômicas e como estas podem impactar no processo de tratamento oncológico. Conclusão: Tais questões são desafiadoras para o trabalho em equipes de saúde, que, diante de situações complexas, precisam desenvolver a interlocução entre diferentes saberes para subsidiar decisões compartilhadas.

Introduction: Cancer is one of the main public health problems in the world. The main risk factors for head and neck cancer are smoking, alcoholism, and others associated with the current economic model, explaining the association between socioeconomic inequalities and incidence/mortality. Objective: To know the socioeconomic profile of people with cancer of the larynx and oral cavity, enrolled in the head and neck clinic at HCI/INCA from February to July 2017. Method: The methodological path used is divided into two axes: the narrative literature review and conceptual definition of terminologies used in the research and the questionnaire applied with sick people. This is an exploratory investigation that aims to raise qualitative elements for the construction of the socioeconomic profile. Results: The results describe the socioeconomic profile, demonstrate the magnitude of socioeconomic conditions and how these can impact the oncology treatment process. Conclusion: These issues are challenging for the work of health teams that, when faced with complex situations, need to develop the dialogue between different types of expertise to support shared decisions.

Introducción: El cáncer es uno de los principales problemas de salud pública en el mundo. El cáncer de cabeza y cuello presenta como principales factores de riesgo el tabaquismo, alcoholismo, entre otros asociados al modelo económico actual, explicando la asociación entre desigualdades socioeconómicas e incidencia/mortalidad. Objetivo: Conocer el perfil socioeconómico de las personas con cáncer de laringe y cavidad oral, inscritas en la clínica de cabeza y cuello del HCI/INCA de febrero a julio de 2017. Método: La ruta metodológica utilizada se divide en dos ejes: la revisión bibliográfica narrativa y definición conceptual de terminologías utilizadas en la investigación y el cuestionario aplicado a las personas enfermas. Se trata de una investigación exploratoria que pretende levantar elementos cualitativos para la construcción del perfil socioeconómico. Resultados: Los resultados presentan el perfil socioeconómico, demuestran la magnitud de las condiciones socioeconómicas y cómo estas pueden impactar en el proceso de tratamiento del cáncer. Conclusión: Tales cuestiones son desafiantes para el trabajo de los equipos de salud que frente a cuestiones complejas necesitan desarrollar la interlocución entre diferentes conocimientos para subvencionar las decisiones compartidas

High-Resolution Magic-Angle-Spinning NMR in Revealing Hepatoblastoma Hallmarks.

Cancer is one of the leading causes of death in children and adolescents worldwide; among the types of liver cancer, hepatoblastoma (HBL) is the most common in childhood. Although it affects only two to three individuals in a million, it is mostly asymptomatic at diagnosis, so by the time it is detected it has already advanced. There are specific recommendations regarding HBL treatment, and ongoing studies to stratify the risks of HBL, understand the pathology, and predict prognostics and survival rates. Although magnetic resonance imaging spectroscopy is frequently used in diagnostics of HBL, high-resolution magic-angle-spinning (HR-MAS) NMR spectroscopy of HBL tissues is scarce. Using this technique, we studied the alterations among tissue metabolites of ex vivo samples from (a) HBL and non-cancer liver tissues (NCL), (b) HBL and adjacent non-tumor samples, and (c) two regions of the same HBL samples, one more centralized and the other at the edge of the tumor. It was possible to identify metabolites in HBL, then metabolites from the HBL center and the border samples, and link them to altered metabolisms in tumor tissues, highlighting their potential as biochemical markers. Metabolites closely related to liver metabolisms such as some phospholipids, triacylglycerides, fatty acids, glucose, and amino acids showed differences between the tissues.

Sensitivity profile of blood culture isolates in a clinical analysis laboratory, Fortaleza, CE / Perfil de sensibilidade de isolados de hemocultura em um laboratório de análises clínicas, Fortaleza, CE / Perfil de sensibilidad de hemocultivos aislados en un laboratorio de análisis clínicos, Fortaleza, CE

Background and objectives: bacteremia is defined from the presence of bacteria in the bloodstream. Its clinical importance is associated with the high morbidity and mortality rate in the world. In severe cases, it can culminate in sepsis, with a constant increase in cases in Brazil. Therefore, this study aims to assess the main bacterial isolates in blood cultures and a possible change in their sensitivity profiles in a clinical analysis laboratory in Fortaleza, Ceará. Methods: an epidemiological, descriptive, retrospective study was carried out, with a quantitative approach of positive blood cultures, seeking to assess the main isolated microorganisms and their sensitivity profiles. The data used were obtained from the laboratory system through the EpiCenter→ software, from January 2019 to December 2020. Statistical analysis was performed using the Graphpad 7.0 software. Results: 840 microorganisms were identified from blood cultures, and the main ones were E. coli, K. pneumoniae, P. aeruginosa, S. epidermidis, S. aureus and S. haemolyticus. Some isolates show a change in the sensitivity profile, such as K. pneumoniae and P. aeruginosa, showing an increase in sensitivity to carbapenems and cephalosporins, while S. epidermidis showed a decrease in sensitivity to minocycline in the comparison between years 2019 and 2020.Conclusion: clinical isolates from blood cultures showed a change in the sensitivity profile between 2019 and 2020, taking into account that, for K. pneumoniae, P. aeruginosa, this change resulted in an increase in sensitivity, with an increase in resistance in S. epidermidis isolates.(AU)

Justificativa e objetivos: bacteremia é definida a partir da presença de bactérias na corrente sanguínea. Sua importância clínica está associada à alta taxa de morbidade e mortalidade no mundo. Nos casos graves, pode culminar em sepse, com constante aumento dos casos no Brasil. Portanto, o presente estudo tem como objetivo avaliar os principais isolados bacterianos em hemoculturas e uma possível alteração nos seus perfis de sensibilidade em um laboratório de análises clínicas de Fortaleza, Ceará. Métodos: foi realizado um estudo epidemiológico, descritivo, retrospectivo, com abordagem quantitativa de hemoculturas positivas, buscando avaliar os principais microrganismos isolados e seus perfis de sensibilidades. Os dados utilizados foram obtidos a partir do sistema laboratorial através do software EpiCenter→, referente ao período de janeiro de 2019 a dezembro de 2020. A análise estatística foi realizada pelo software Graphpad 7.0. Resultados: foram identificados 840 microrganismos a partir das hemoculturas, sendo os principais E. coli, K. pneumoniae, P. aeruginosa, S. epidermidis, S. aureus e S. haemolyticus. Alguns isolados apresentam uma alteração no perfil de sensibilidade, como K. pneumoniae e P. aeruginosa, apresentando um aumento na sensibilidade frente aos carbapenêmicos e as cefalosporinas, enquanto o S. epidermidis apresentou uma diminuição na sensibilidade frente à minociclina na comparação entre os anos de 2019 e 2020. Conclusão: os isolados clínicos de hemocultura apresentaram uma alteração no perfil de sensibilidade entre 2019 e 2020, levando em consideração que, para K. pneumoniae e P. aeruginosa, essa alteração resultou no aumento na sensibilidade, com aumento na resistência nos isolados de S. epidermidis.(AU)

Justificación y objetivos: la bacteriemia se define por la presencia de bacterias en el torrente sanguíneo. Su importancia clínica está asociada con la alta tasa de morbimortalidad en el mundo. En casos severos, puede culminar en sepsis, con un aumento constante de casos en Brasil. Por tanto, este estudio tiene como objetivo evaluar los principales aislados bacterianos en hemocultivos y un posible cambio en sus perfiles de sensibilidad en un laboratorio de análisis clínicos en Fortaleza, Ceará. Métodos: se realizó un estudio epidemiológico, descriptivo, retrospectivo, con abordaje cuantitativo de hemocultivos positivos, buscando evaluar los principales microorganismos aislados y sus perfiles de sensibilidad. Los datos utilizados se obtuvieron del sistema de laboratorio a través del software EpiCenter→, para el período de enero de 2019 a diciembre de 2020. El análisis estadístico se realizó mediante el software Graphpad 7.0. Resultados: se identificaron 840 microorganismos a partir de hemocultivos, siendo los principales E. coli, K. pneumoniae, P. aeruginosa, S. epidermidis, S. aureus y S. haemolyticus. Algunos aislados muestran un cambio en el perfil de sensibilidad, como K.pneumoniae y P. aeruginosa, mostrando un aumento en la sensibilidad a los carbapenémicos y cefalosporinas, mientras que S. epidermidis mostró una disminución en la sensibilidad a la minociclina, en la comparación entre los años de 2019 y 2020. Conclusiones: los aislados clínicos de hemocultivos mostraron un cambio en el perfil de sensibilidad entre 2019 y 2020, teniendo en cuenta que para K. pneumoniae, P. aeruginosa, este cambio resultó en un aumento de la sensibilidad, con un aumento de la resistencia en los aislados de S. epidermidis

An ion imprinted magnetic organosilica nanocomposite for the selective determination of traces of Cd(II) in a minicolumn flow-through preconcentration system coupled with graphite furnace atomic absorption spectroscopy.

In this paper we present the determination of ultratraces of cadmium ions in water by means of a minicolumn (MC) flow-through preconcentration system coupled with graphite furnace atomic absorption spectrometry. The core of the system is a lab-made ion imprinted magnetic organosilica nanocomposite which is employed as filler of the MC for the selective retention of the analyte. In this case superparamagnetic magnetite nanoparticles were coated with an amine-functionalized shell and ion imprinted with Cd(II) by a simple sol-gel co-condensation method. The setup was completed with the inclusion of a magnet fixed around the packed MC. This assembly - which is studied with an MII material for the first time here - allowed a homogeneous distribution of the solid on the walls of the MC, leaving a hole in the center and enabling the absence of material bleeding or obstructions to the free movement of fluids. Ion imprinted (MII) and non-imprinted (MNI) materials were studied for comparison purposes. Both were characterized and compared by DRX, FTIR, and SEM and their magnetic behavior by magnetization curves. Batch experiments showed an equilibration time of less than 10 minutes and a maximum adsorption pH of around 7 for both solids. The maximum capacity for MII was greater than that of MNI (200 mg g-1 and 30 mg g-1 respectively) and thus, the former was chosen for analytical purposes. Under MC dynamic conditions, sample and elution flow rates, volumes of the sample and eluant, and type and concentration of the most suitable eluant have been thoroughly investigated and optimized. Under the optimal experimental conditions, the MII filler showed a preconcentration factor of 200, a limit of detection of 0.64 ng L-1, a linear range of 2.5-100 ng L-1, RSD% of 1.9 (n = 6; 10 ng L-1) and a lifetime of more than 800 cycles of concentration-elution with no loss of sensitivity or need for refilling. The effect of potentially interfering ions on the percent recovery of cadmium was also studied. The proposed method was successfully applied to the determination of traces of Cd(II) in osmosis and tap water with recoveries of 98.0-101.3%. A comparison with similar methods is also provided.

Antimicrobial susceptibility and production of virulence factors by bacteria recovered from bitches with pyometra.

Pyometra is one of the most common diseases in adult female dogs, characterized by a suppurative bacterial infection of the uterus with accumulation of inflammatory exudate and a variety of local and systemic clinical manifestations. This study aimed to identify the bacteria within the uterine content and vaginal canal of bitches with pyometra and evaluate their antimicrobial susceptibility and production of virulence factors. Uterine and vaginal content were collected with sterile swabs from 30 bitches diagnosed with pyometra. Bacteria were identified and assessed for their antimicrobial susceptibility and production of virulence factors, including biofilms, siderophores, proteases and hemolysins, both in planktonic and biofilm forms. A total of 82 bacterial isolates (35 uterus, 47 vagina), belonging to 21 species, were identified, with Escherichia coli as the most prevalent species (32/82, 39%). As for susceptibility, 39/79 (49.4%) isolates were resistant to one or more drugs, with resistance proportion among Gram-positive bacteria (87.5%) higher (p < .05) than that observed for Gram-negative bacteria (32.7%). Four coagulase-negative Staphylococcus species were resistant to methicillin. Regarding virulence, the isolates had low production of biofilms, siderophores, proteases and hemolysins, suggesting that the occurrence of pyometra might be more associated with host-related factors than bacterial virulence.

Unraveling the Genetic Architecture of Hepatoblastoma Risk: Birth Defects and Increased Burden of Germline Damaging Variants in Gastrointestinal/Renal Cancer Predisposition and DNA Repair Genes.

The ultrarare hepatoblastoma (HB) is the most common pediatric liver cancer. HB risk is related to a few rare syndromes, and the molecular bases remain elusive for most cases. We investigated the burden of rare damaging germline variants in 30 Brazilian patients with HB and the presence of additional clinical signs. A high frequency of prematurity (20%) and birth defects (37%), especially craniofacial (17%, including craniosynostosis) and kidney (7%) anomalies, was observed. Putative pathogenic or likely pathogenic monoallelic germline variants mapped to 10 cancer predisposition genes (CPGs: APC, CHEK2, DROSHA, ERCC5, FAH, MSH2, MUTYH, RPS19, TGFBR2 and VHL) were detected in 33% of the patients, only 40% of them with a family history of cancer. These findings showed a predominance of CPGs with a known link to gastrointestinal/colorectal and renal cancer risk. A remarkable feature was an enrichment of rare damaging variants affecting different classes of DNA repair genes, particularly those known as Fanconi anemia genes. Moreover, several potentially deleterious variants mapped to genes impacting liver functions were disclosed. To our knowledge, this is the largest assessment of rare germline variants in HB patients to date, contributing to elucidate the genetic architecture of HB risk.

Clostridioides difficile from Brazilian hospitals: characterization of virulence genes by whole genome sequencing.

Clostridioides difficile (CD) is the most frequent cause of healthcare related diarrhea and its severity has increased in the last decade by the spread of hypervirulent strains. Most important CD virulence factor is toxin production; however, not only toxins are responsible for Clostridioides virulence. We sequenced 38 strains and analyzed the presence and integrity of 24 virulence (including toxin) genes. We identified 28 toxigenic strains, six also presented the cdt genes. Only six strains didn't present all others genes searched. All absent genes were adhesion related. Understand others CD virulence factors can lead to a best understanding on this matter.

Comparative biofilm-forming ability between Clostridioides difficile strains isolated in Latin America and the epidemic NAP1/027 strain.

Introduction: One of the challenges in treating Clostridioides difficile infection (CDI) is that the bacterium forms biofilms, a critical virulence mechanism known to promote antibiotic resistance and, as a result, consequently, a higher recurrence of the disease. The goal of this study was to compare the ability of three MLST Clade 2 strains to form a biofilm in vitro: ICC-45 (ribotype SLO231/UK[CE]821), a ST41 toxinotype IXb isolated in Brazil; and two epidemic NAP1/027/ST01 strains: NAP1/027/ST01 (LIBA5756), isolated during a 2010 outbreak in Costa Rica and the reference epidemic strain NAP1/027/ST01 (R20291); and ATCC700057, a non-toxigenic strain. Methods: The ability of strains to form biofilm was evaluated using crystal violet staining. In addition, samples were stained with the Film Tracer biofilm matrix (Invitrogen®) and the biofilm matrix thickness was measured using confocal microscopy. The matrix architecture was determined using Scanning electron microscop. Confocal microscopy was used to detect the presence of toxin A (tcdA) using an anti-Clostridioides difficile TcdA antibody. The expression of virulence genes (tcdA, tcdB, tcdC, cdtB, spo0A, slpA, cwp66 and cwp84) was examined, as well as the effect of antibiotics metronidazole (MTZ) and vancomycin (VAN) on biofilm growth. Results: All of the strains tested formed a moderate biofilm with 1.1 3.5. After 72h, biofilm biomass of the NAP1/027/ST01 epidemic strains (LIBA5756 and R20291) was significantly higher than ICC-45 and ATCC 700057 biofilms, as confirmed by electron and confocal microscopy. At 120h, the LIBA5756 biofilm biomass decreased compared to other strains. The toxigenic strains R20291 or LIBA 5756 had higher expression of genes tcdA, tcdB, tcdC, cdtA, slpA and spo0A than ICC-45, but there were no significant differences in the expression levels of cdtB, cwp66 and cwp84. In epidemic strains, VAN and MTZ inhibited biofilm formation; however, in the ICC-45 strain, MIC concentrations of VAN and MIC and 4MIC of MTZ did not inhibit biofilm formation. Conclusion: The three MLST Clade 2 isolated from different rybotipes, two of which were isolated from Latin America, are competent biofilm-forming bacteria, indicating their ability to induce C. difficile infection recurrence, making treatment difficult.

Preserved executive functioning and low stress symptoms in children treated for acute lymphoblastic leukemia.

Pediatric cancer treatment can negatively impact cognitive and psychosocial development, although it has been suggested that these adverse effects may be minimized when children have higher resilience and better executive functioning. We aimed to evaluate the impact of pediatric Acute Lymphoblastic Leukemia (ALL) treatment on executive function, resilience and stress in survivors and to investigate correlations between executive functioning and resilience and between executive functioning and stress. The neuropsychological assessment was performed in 32 ALL survivors aged 7-17 years and 28 age-, sex- and socioeconomic status matched controls. Executive functioning was assessed by inhibitory control, mental flexibility and working memory tasks. Children's self-report scales were used to assess stress symptoms and resilience. Results revealed no executive function impairment nor stress symptom differences between ALL survivors and control group. In the ALL group, executive function and resilience were positively correlated, whereas executive function and stress were negatively correlated. We concluded that ALL treatment was not associated with impairment in executive functioning nor to increased stress symptoms in our sample. ALL survivors with better performance in mental flexibility and inhibition tasks reported fewer stress symptoms and more resilience, indicating a possible relationship between these variables.

Copy Number Alterations in Hepatoblastoma: Literature Review and a Brazilian Cohort Analysis Highlight New Biological Pathways.

Hepatoblastoma (HB) is a rare embryonal tumor, although it is the most common pediatric liver cancer. The aim of this study was to provide an accurate cytogenomic profile of this type of cancer, for which information in cancer databases is lacking. We performed an extensive literature review of cytogenetic studies on HBs disclosing that the most frequent copy number alterations (CNAs) are gains of 1q, 2/2q, 8/8q, and 20; and losses at 1p and 4q. Furthermore, the CNA profile of a Brazilian cohort of 26 HBs was obtained by array-CGH; the most recurrent CNAs were the same as shown in the literature review. Importantly, HBs from female patients, high-risk stratification tumors, tumors who developed in older patients (> 3 years at diagnosis) or from patients with metastasis and/or deceased carried a higher diversity of chromosomal alterations, specifically chromosomal losses at 1p, 4, 11q and 18q. In addition, we distinguished three major CNA profiles: no detectable CNA, few CNAs and tumors with complex genomes. Tumors with simpler genomes exhibited a significant association with the epithelial fetal subtype of HBs; in contrast, the complex genome group included three cases with epithelial embryonal histology, as well as the only HB with HCC features. A significant association of complex HB genomes was observed with older patients who developed high-risk tumors, metastasis, and deceased. Moreover, two patients with HBs exhibiting complex genomes were born with congenital anomalies. Together, these findings suggest that a high load of CNAs, mainly chromosomal losses, particularly losses at 1p and 18, increases the tendency to HB aggressiveness. Additionally, we identified six hot-spot chromosome regions most frequently affected in the entire group: 1q31.3q42.3, 2q23.3q37.3, and 20p13p11.1 gains, besides a 5,3 Mb amplification at 2q24.2q24.3, and losses at 1p36.33p35.1, 4p14 and 4q21.22q25. An in-silico analysis using the genes mapped to these six regions revealed several enriched biological pathways such as ERK Signaling, MicroRNAs in Cancer, and the PI3K-Akt Signaling, in addition to the WNT Signaling pathway; further investigation is required to evaluate if disturbances of these pathways can contribute to HB tumorigenesis. The analyzed gene set was found to be associated with neoplasms, abnormalities of metabolism/homeostasis and liver morphology, as well as abnormal embryonic development and cytokine secretion. In conclusion, we have provided a comprehensive characterization of the spectrum of chromosomal alterations reported in HBs and identified specific genomic regions recurrently altered in a Brazilian HB group, pointing to new biological pathways, and relevant clinical associations.

Infection with Clostridioides difficile ribotype 046 in a paediatric liver transplant patient.

Clostridioides difficile causes nosocomial diarrhoea associated with antibiotic use and immunodeficiency. Although the number of paediatric C. difficile infections (CDIs) has increased worldwide, there are few studies on the molecular characterization of strains causing CDIs among children. We report the clinical features and strain molecular characterization of a CDI in a female child with a history of liver transplantation at 7 months of age. This is the first report of the 046 ribotype causing paediatric diarrhoea.

Exoproteomic analysis of two MLST clade 2 strains of Clostridioides difficile from Latin America reveal close similarities.

Clostridioides difficile BI/NAP1/ribotype 027 is an epidemic hypervirulent strain found worldwide, including in Latin America. We examined the genomes and exoproteomes of two multilocus sequence type (MLST) clade 2 C. difficile strains considered hypervirulent: ICC-45 (ribotype SLO231/UK[CE]821), isolated in Brazil, and NAP1/027/ST01 (LIBA5756), isolated during a 2010 outbreak in Costa Rica. C. difficile isolates were cultured and extracellular proteins were analyzed using high-performance liquid chromatography-tandem mass spectrometry. Genomic analysis revealed that these isolates shared most of the gene composition. Only 83 and 290 NAP1/027 genes were considered singletons in ICC-45 and NAP1/027, respectively. Exoproteome analysis revealed 197 proteins, of which 192 were similar in both strains. Only five proteins were exclusive to the ICC-45 strain. These proteins were involved with catalytic and binding functions and indirectly interacted with proteins related to pathogenicity. Most proteins, including TcdA, TcdB, flagellin subunit, and cell surface protein, were overrepresented in the ICC-45 strain; 14 proteins, including mature S-layer protein, were present in higher proportions in LIBA5756. Data are available via ProteomeXchange with identifier PXD026218. These data show close similarity between the genome and proteins in the supernatant of two strains with hypervirulent features isolated in Latin America and underscore the importance of epidemiological surveillance of the transmission and emergence of new strains.