Saprochaete clavata invasive infection: characterization, antifungal susceptibility, and biofilm evaluation of a rare yeast isolated in Brazil

ABSTRACT Rare emerging pathogens such as Saprochaete clavata are associated with invasive fungal diseases, high morbidity, mortality, rapidly fatal infections, and outbreaks. However, little is known about S. clavata infections, epidemiology, risk factors, treatment, biofilms, and disease outcomes. The objective of this study was to describe a new case of severe S. clavata infection in a patient diagnosed at a referral children's hospital in Brazil, including antifungal minimal inhibitory concentration, S. clavata biofilm characterization, and molecular characterization. The S. clavata isolated from an immunocompromised 11-year-old male patient was characterized using MALDI-TOF, Gram staining, scanning electron microscopy (SEM), and next generation sequencing (NGS) of genomic DNA. Biofilm production was also evaluated in parallel with determining minimal inhibitory concentration (MIC) and biofilm sensitivity to antifungal treatment. We observed small to medium, whitish, farinose, dry, filamentous margin colonies, yeast-like cells with bacillary features, and biofilm formation. The MALDI-TOF system yielded a score of ≥ 2,000, while NGS confirmed S. clavata presence at the nucleotide level. The MIC values (in mg L-1) for tested drugs were as follows: fluconazole = 2, voriconazole ≤ 2, caspofungin ≥ 8, micafungin = 2, amphotericin B = 4, flucytosine ≤ 1, and anidulafungin = 1. Amphotericin B can be active against S. clavata biofilm and the fungus can be susceptible to new azoles. These findings were helpful for understanding the development of novel treatments for S. clavata-induced disease, including combined therapy for biofilm-associated infections.

Systems biology network reveals the correlation between COX-2 expression and Ch 7q copy number alterations in Ch 11q-deleted pediatric neuroblastoma tumors.

Tumor-associated inflammation and chromosomal aberrations can play crucial roles in cancer development and progression. In neuroblastoma (NB), the enzyme cyclooxygenase-2 (COX-2) is associated with copy number alterations on the long arm of chromosome 11 (Ch 11q), defining an aggressive disease subset. This retrospective study included formalin-fixed paraffin-embedded tumor samples collected from nine patients during diagnosis at the pediatric Pequeno Principe Hospital, Curitiba, PR, Brazil, and post-chemotherapy (CT). COX-2 expression was evaluated using immunohistochemistry and correlated with the genome profile of paired pre- and post-CT samples, determined by array comparative genomic hybridization. A systems biology approach elucidated the PTGS2 network interaction. The results showed positive correlations between pre-CT Ch 7q gain and COX-2 expression (ρ = 0.825; p-value = 0.006) and negative correlations between Ch 7q gain and Ch 11q deletion (ρ = -0.919; p-value = 0.0005). Three samples showed Ch 11q deletion and Ch 7q gain. Network analysis identified a direct connection between CAV-1 (Ch 7q) and COX-2 in NB tumors and highlighted the connection between amplified genes in Ch 7q and deleted ones in 11q. The identification of hub-bottleneck-switch genes provides new biological insights into this connection between NB, tumorigenesis, and inflammation.

A comprehensive analysis of AHRR gene as a candidate for cleft lip with or without cleft palate.

Cleft lip and palate (CL/P) is among the most common congenital malformations and affects 1 in 700 newborns. CL/P is caused by genetic and environmental factors (maternal smoking, alcohol or drug use and others). Many genes and loci were associated with cleft lip/palate but the amount of heterogeneity justifies identifying new causal genes and variants. AHRR (Aryl-Hydrocarbon Receptor Repressor) gene has recently been related to CL/P however, few functional studies analyze the genotypephenotype interaction of AHRR with CL/P. Several studies associate the molecular pathway of AHRR to CL/P which indicates this gene as a functional candidate in CL/P etiology. METHODS: Systematic Literature Review was performed using PUBMED database with the keywords cleft lip, cleft palate, orofacial cleft, AHRR and synonyms. SLR resulted in 37 included articles. RESULTS: AHRR is a positional and functional candidate gene for CL/P. In silico analysis detected interactions with other genes previously associated to CL/P like ARNT and CYP1A1. AHRR protein regulates cellular toxicity through TCDD mediated AHR pathway. Exposure to TCDD in animal embryos is AHR mediated and lead to cleft palate due to palate fusion failure and post fusion rupture. AHRR regulates cellular growth and differentiation, fundamental to lip and palatogenesis. AHRR decreases carcinogenesis and recently a higher tumor risk has been described in CL/P patients and families. AHRR is also a smoking biomarker due to changed methylation sites found in smokers DNA although folate intake may partially revert these methylation alterations. This corroborates the role of maternal smoking and lack of folate supplementation as risk factors for CL/P. CONCLUSION: This research identified the importance of AHRR in dioxin response and demonstrated an example of genetic and environmental interaction, indispensable in the development of many complex diseases.

Avaliação do perfil lipídico de adolescentes / Evaluation of lipid profile in adolescents

A aterosclerose é uma doença crônica, multifatorial e insidiosa, podendo iniciar-se na infância ou adolescência, com suas principais consequências aparecendo na fase adulta. As dosagens séricas de lipoproteínas como LDL-c, colesterol total (CT), HDL-c e NÃO HDL-c podem ser usadas como forma de triagem de um diagnóstico. No Brasil há ainda pouquíssimos estudos correlacionando níveis séricos dessas lipoproteínas com a idade das pessoas. Avaliar as concentrações séricas de LDL-c, CT, HDL-c, NÃO HDL-c, VLDL-c e triglicerídeos (TG) em adolescentes de 10 a 19 anos do município de Araucária/PR. Pesquisa transversal retrospectiva, que coletou os seguintes dados de 600 adolescentes: idade, sexo e dosagens de LDL-c, CT, HDL-c, NÃO HDL-c, VLDL-c e TG. Os dados foram avaliados com o programa de análise estatística SPSS 2.0, o teste U de Mann-Whitney e o coeficiente de correlação de Spearman para identificação de significado estatístico (p < 0,05). O sexo feminino exprimiu níveis séricos de CT, TG e LDL-c maiores que o sexo masculino. O HDL-c apresentou valores idênticos em ambos os sexos, com 48% de valores desejáveis e 52% de baixos. O estudo identificou forte correlação entre as frações lipídicas e associação com a idade de 10 a 14 anos. Os resultados apontam que, em comparação ao LDL-c, o NÃO HDL-c apresentou maior correlação com as demais frações lipídicas (TG, LDL-c e CT), sugerindo que o NÃO HDL-c pode ser utilizado como um método eficaz na complementação de diagnóstico para avaliar riscos ateroscleróticos em adolescentes

Atherosclerosis is a chronic, multifactorial and insidious disease that can begin in childhood and adolescence, and whose major consequences appear during adulthood. Serum levels of lipoproteins, such as LDL-c, total cholesterol (TC), HDL-c, and non-HDL-c can be used as a screening method for disease diagnosis. In Brazil, few studies have correlated the serum levels of those lipoproteins with age. To evaluate the serum concentrations of TC, LDL-c, HDL-c, VLDL-c, non-HDL-c and triglycerides (TG) of adolescents aged 10 to 19 years in the municipality of Araucária, Paraná state. Cross-sectional retrospective study, collecting the following data from 600 adolescents: age, sex and serum levels of TC, LDL-c, HDL-c and TG from June to December 2016. Data were analyzed using the SPSS software 2.0, with Mann-Whitney U test and Spearman coefficient of correlation to identify statistical significance (p < 0.05). The female sex showed higher serum levels of TC, TG and LDL-c than the male sex. The HDL-c levels were identical in both sexes, with 48% of desirable values and 52% of low values. This study identified a strong correlation between the lipids and association with the age group of 10 to 14 years. Non-HDL-c showed stronger correlation with the other lipids (TG, LDL-c and TC) as compared to LDL-c, suggesting that non-HDL-c can be used as an effective complementary diagnostic method to assess the risks for atherosclerosis in adolescents

A quantitative proteomic and transcriptomic comparison of human mesenchymal stem cells from bone marrow and umbilical cord vein.

Human mesenchymal stem cells (hMSCs) are adult multipotent cells that have high therapeutic potential due to their immunological properties. They can be isolated from several different tissues with bone marrow (BM) being the most common source. Because the isolation procedure is invasive, other tissues such as human umbilical cord vein (UCV) have been considered. However, their interchangeability remains unclear. In the present study, total protein extracts of BM-hMSCs and UCV-hMSCs were quantitatively compared using gel-LC-MS/MS. Previous SAGE analysis of the same cells was re-annotated to enable comparison and combination of these two data sets. We observed a more than 63% correlation between proteomic and transcriptomic data. In silico analysis of highly expressed genes in cells of both origins suggests that they can be modulated by microRNA, which can change protein abundance. Our results showed that MSCs from both tissues shared high similarity in metabolic and functional processes relevant to their therapeutic potential, especially in the immune system process, response to stimuli, and processes related to the delivery of the hMSCs to a given tissue, such as migration and adhesion. Hence, our results support the idea that the more accessible UCV could be a potentially less invasive source of MSCs.

Detection of human interchromosomal trans-splicing in sequence databanks.

Trans-splicing is a common phenomenon in nematodes and kinetoplastids, and it has also been reported in other organisms, including humans. Up to now, all in silico strategies to find evidence of trans-splicing in humans have required that the candidate sequences follow the consensus splicing site rules (spliceosome-mediated mechanism). However, this criterion is not supported by the best human experimental evidence, which, except in a single case, do not follow canonical splicing sites. Moreover, recent findings describe a novel alternative tRNA mediated trans-splicing mechanism, which prescinds the spliceosome machinery. In order to answer the question, 'Are there hybrid mRNAs in sequence databanks, whose characteristics resemble those of the best human experimental evidence?', we have developed a methodology that successfully identified 16 hybrid mRNAs which might be instances of interchromosomal trans-splicing. Each hybrid mRNA is formed by a trans-spliced region (TSR), which was successfully mapped either onto known genes or onto a human endogenous retrovirus (HERV-K) transcript which supports their transcription. The existence of these hybrid mRNAs indicates that trans-splicing may be more widespread than believed. Furthermore, non-canonical splice site patterns suggest that infrequent splicing sites may occur under special conditions, or that an alternative trans-splicing mechanism is involved. Finally, our candidates are supposedly from normal tissue, and a recent study has reported that trans-splicing may occur not only in malignant tissues, but in normal tissues as well. Our methodology can be applied to 5'-UTR, coding sequences and 3'-UTR in order to find new candidates for a posteriori experimental confirmation.