BUB1B monoallelic germline variants contribute to prostate cancer predisposition by triggering chromosomal instability.

BACKGROUND: Prostate cancer (PrCa) is the most frequently diagnosed cancer in men. Variants in known moderate- to high-penetrance genes explain less than 5% of the cases arising at early-onset (< 56 years) and/or with familial aggregation of the disease. Considering that BubR1 is an essential component of the mitotic spindle assembly checkpoint, we hypothesized that monoallelic BUB1B variants could be sufficient to fuel chromosomal instability (CIN), potentially triggering (prostate) carcinogenesis. METHODS: To unveil BUB1B as a new PrCa predisposing gene, we performed targeted next-generation sequencing in germline DNA from 462 early-onset/familial PrCa patients and 1,416 cancer patients fulfilling criteria for genetic testing for other hereditary cancer syndromes. To explore the pan-cancer role of BUB1B, we used in silico BubR1 molecular modeling, in vitro gene-editing, and ex vivo patients' tumors and peripheral blood lymphocytes. RESULTS: Rare BUB1B variants were found in ~ 1.9% of the early-onset/familial PrCa cases and in ~ 0.6% of other cancer patients fulfilling criteria for hereditary disease. We further show that BUB1B variants lead to decreased BubR1 expression and/or stability, which promotes increased premature chromatid separation and, consequently, triggers CIN, driving resistance to Taxol-based therapies. CONCLUSIONS: Our study shows that different BUB1B variants may uncover a trigger for CIN-driven carcinogenesis, supporting the role of BUB1B as a (pan)-cancer predisposing gene with potential impact on genetic counseling and treatment decision-making.

Genetic landscape of homologous recombination repair genes in early-onset/familial prostate cancer patients.

Prostate cancer (PrCa) is one of the three most frequent and deadliest cancers worldwide. The discovery of PARP inhibitors for the treatment of tumors with deleterious variants in homologous recombination repair (HRR) genes has placed PrCa on the roadmap of precision medicine. However, the overall contribution of HRR genes to the 10%-20% of carcinomas arising in men with early-onset/familial PrCa has not been fully clarified. We used targeted next-generation sequencing (T-NGS) covering eight HRR genes (ATM, BRCA1, BRCA2, BRIP1, CHEK2, NBN, PALB2, and RAD51C) and an analysis pipeline querying both small and large genomic variations to clarify their global and relative contribution to hereditary PrCa predisposition in a series of 462 early-onset/familial PrCa cases. Deleterious variants were found in 3.9% of the patients, with CHEK2 and ATM being the most frequently mutated genes (38.9% and 22.2% of the carriers, respectively), followed by PALB2 and NBN (11.1% of the carriers, each), and finally by BRCA2, RAD51C, and BRIP1 (5.6% of the carriers, each). Using the same NGS data, exonic rearrangements were found in two patients, one pathogenic in BRCA2 and one of unknown significance in BRCA1. These results contribute to clarify the genetic heterogeneity that underlies PrCa predisposition in the early-onset and familial disease, respectively.

Exome sequencing of affected duos and trios uncovers PRUNE2 as a novel prostate cancer predisposition gene.

BACKGROUND: Prostate cancer (PrCa) is one of the most hereditable human cancers, however, only a small fraction of patients has been shown to carry deleterious variants in known cancer predisposition genes. METHODS: Whole-exome sequencing was performed in multiple affected members of 45 PrCa families to select the best candidate genes behind part of the PrCa missing hereditability. Recurrently mutated genes were prioritised, and further investigated by targeted next-generation sequencing in the whole early-onset and/or familial PrCa series of 462 patients. RESULTS: PRUNE2 stood out from our analysis when also considering the available data on its association with PrCa development. Ten germline pathogenic/likely pathogenic variants in the PRUNE2 gene were identified in 13 patients. The most frequent variant was found in three unrelated patients and identical-by-descent analysis revealed that the haplotype associated with the variant is shared by all the variant carriers, supporting the existence of a common ancestor. DISCUSSION: This is the first report of pathogenic/likely pathogenic germline variants in PRUNE2 in PrCa patients, namely in those with early-onset/familial disease. Importantly, PRUNE2 was the most frequently mutated gene in the whole series, with a deleterious germline variant identified in 2.8% of the patients, representing a novel prostate cancer predisposition gene.

Reaching Men and Young Adults in a Pharmacy-Based HIV Self-Testing Strategy: Results from an Acceptability Study in Mozambique.

HIV self-testing (HIVST) is a WHO-recommended strategy to increase testing, especially among key populations, men, and young adults. Between May and December 2019, a pilot was implemented in Zambézia province, Mozambique, allowing clients to purchase HIV self-tests in 14 public/private pharmacies. The study assessed the strategy's acceptability and uptake. Pharmacy-based exit surveys were conducted among a random sample of clients, during the first three months of the pilot, independent of HIVST purchase. Another random sample of clients who bought an HIVST completed a survey 1-12 weeks after purchase. Chi-square and Mann-Whitney tests were used for the analysis, comparing clients who purchased an HIVST versus not. A total of 1,139 adults purchased 1,344 tests. Buyers were predominantly male (70%) and younger (52% between 15 and 34 years of age). Surveys were completed by 280 exiting pharmacy clients and 82 clients who purchased an HIVST. Main advantages were confidentiality and lack of need of a health provider visit, with main disadvantages being absence of nearby counseling and fear of results. No differences were seen between buyers and non-buyers for these factors. Among all undergoing HIVST, 71 (92%) perceived the instructions to be clear, however, 29 (38%) stated they would have benefitted from additional pre-test information or counseling. Ten (13%) reported following up at a nearby health facility to confirm results and/or receive care. Offering HIVST at public/private pharmacies was acceptable among people who traditionally tend to have a lower HIV testing coverage, such as men and young adults. However, additional resources and/or enhanced educational materials to address the lack of counseling, and linkage-to-care systems need to be put into place before scaling up this strategy.

Glycoproteogenomics characterizes the CD44 splicing code associated with bladder cancer invasion.

Rationale: Bladder cancer (BC) management demands the introduction of novel molecular targets for precision medicine. Cell surface glycoprotein CD44 has been widely studied as a potential biomarker of BC aggressiveness and cancer stem cells. However, significant alternative splicing and multiple glycosylation generate a myriad of glycoproteoforms with potentially distinct functional roles. The lack of tools for precise molecular characterization has led to conflicting results, delaying clinical applications. Addressing these limitations, we have interrogated the transcriptome and glycoproteome of a large BC patient cohort for splicing signatures. Methods:CD44 gene and its splicing variants were assessed by Real Time-Polymerase Chain Reaction (RT-PCR) and RNAseq in tumor tissues. The co-localization of CD44 and short O-glycans was evaluated by proximity ligation assay (PLA), immunohistochemistry and double-immunofluorescence. An innovative glycoproteogenomics approach, integrating transcriptomics-customized datasets and glycomics for protein annotation from nanoLC-ESI-MS/MS experiments, was developed and implemented to identify CD44 variants and associated glycosignatures. The impact of CD44 silencing on proliferation and invasion of BC cell lines and glycoengineered cells was determined by BrdU ELISA and Matrigel invasion assays, respectively. Antibody phosphoarrays were used to investigate the role of CD44 and its glycoforms in the activation of relevant oncogenic signaling pathways. Results: Transcriptomics analysis revealed remarkable CD44 isoforms heterogeneity in bladder cancer tissues, as well as associations between short CD44 standard splicing isoform (CD44s), invasion and poor prognosis. We further demonstrated that targeting short O-glycoforms such as the Tn and sialyl-Tn antigens was key to overcome the lack of cancer specificity presented by CD44. Glycoproteogenomics allowed, for the first time, the comprehensive characterization of CD44 splicing code at the protein level. The concept was applied to invasive human BC cell lines, glycoengineered cells, and tumor tissues, enabling unequivocal CD44s identification as well as associated glycoforms. Finally, we confirmed the link between CD44 and invasion in CD44s-enriched cells in vitro by small interfering RNA (siRNA) knockdown, supporting findings from BC tissues. The key role played by short-chain O-glycans in CD44-mediated invasion was also demonstrated through glycoengineered cell models. Conclusions: Overall, CD44s emerged as biomarker of poor prognosis and CD44-Tn/ Sialyl-Tn (STn) as promising molecular signatures for targeted interventions. This study materializes the concept of glycoproteogenomics and provides a key vision to address the cancer splicing code at the protein level, which may now be expanded to better understand CD44 functional role in health and disease.

Glycoproteomics identifies HOMER3 as a potentially targetable biomarker triggered by hypoxia and glucose deprivation in bladder cancer.

BACKGROUND: Muscle invasive bladder cancer (MIBC) remains amongst the deadliest genitourinary malignancies due to treatment failure and extensive molecular heterogeneity, delaying effective targeted therapeutics. Hypoxia and nutrient deprivation, oversialylation and O-glycans shortening are salient features of aggressive tumours, creating cell surface glycoproteome fingerprints with theranostics potential. METHODS: A glycomics guided glycoproteomics workflow was employed to identify potentially targetable biomarkers using invasive bladder cancer cell models. The 5637 and T24 cells O-glycome was characterized by mass spectrometry (MS), and the obtained information was used to guide glycoproteomics experiments, combining sialidase, lectin affinity and bottom-up protein identification by nanoLC-ESI-MS/MS. Data was curated by a bioinformatics approach developed in-house, sorting clinically relevant molecular signatures based on Human Protein Atlas insights. Top-ranked targets and glycoforms were validated in cell models, bladder tumours and metastases by MS and immunoassays. Cells grown under hypoxia and glucose deprivation disclosed the contribution of tumour microenvironment to the expression of relevant biomarkers. Cancer-specificity was validated in healthy tissues by immunohistochemistry and MS in 20 types of tissues/cells of different individuals. RESULTS: Sialylated T (ST) antigens were found to be the most abundant glycans in cell lines and over 900 glycoproteins were identified potentially carrying these glycans. HOMER3, typically a cytosolic protein, emerged as a top-ranked targetable glycoprotein at the cell surface carrying short-chain O-glycans. Plasma membrane HOMER3 was observed in more aggressive primary tumours and distant metastases, being an independent predictor of worst prognosis. This phenotype was triggered by nutrient deprivation and concomitant to increased cellular invasion. T24 HOMER3 knockdown significantly decreased proliferation and, to some extent, invasion in normoxia and hypoxia; whereas HOMER3 knock-in increased its membrane expression, which was more pronounced under glucose deprivation. HOMER3 overexpression was associated with increased cell proliferation in normoxia and potentiated invasion under hypoxia. Finally, the mapping of HOMER3-glycosites by EThcD-MS/MS in bladder tumours revealed potentially targetable domains not detected in healthy tissues. CONCLUSION: HOMER3-glycoforms allow the identification of patients' subsets facing worst prognosis, holding potential to address more aggressive hypoxic cells with limited off-target effects. The molecular rationale for identifying novel bladder cancer molecular targets has been established.

Rare Germline Variants in ATM Predispose to Prostate Cancer: A PRACTICAL Consortium Study.

BACKGROUND: Germline ATM mutations are suggested to contribute to predisposition to prostate cancer (PrCa). Previous studies have had inadequate power to estimate variant effect sizes. OBJECTIVE: To precisely estimate the contribution of germline ATM mutations to PrCa risk. DESIGN, SETTING, AND PARTICIPANTS: We analysed next-generation sequencing data from 13 PRACTICAL study groups comprising 5560 cases and 3353 controls of European ancestry. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Variant Call Format files were harmonised, annotated for rare ATM variants, and classified as tier 1 (likely pathogenic) or tier 2 (potentially deleterious). Associations with overall PrCa risk and clinical subtypes were estimated. RESULTS AND LIMITATIONS: PrCa risk was higher in carriers of a tier 1 germline ATM variant, with an overall odds ratio (OR) of 4.4 (95% confidence interval [CI]: 2.0-9.5). There was also evidence that PrCa cases with younger age at diagnosis (<65 yr) had elevated tier 1 variant frequencies (pdifference = 0.04). Tier 2 variants were also associated with PrCa risk, with an OR of 1.4 (95% CI: 1.1-1.7). CONCLUSIONS: Carriers of pathogenic ATM variants have an elevated risk of developing PrCa and are at an increased risk for earlier-onset disease presentation. These results provide information for counselling of men and their families. PATIENT SUMMARY: In this study, we estimated that men who inherit a likely pathogenic mutation in the ATM gene had an approximately a fourfold risk of developing prostate cancer. In addition, they are likely to develop the disease earlier.

The spread of visceral leishmaniasis in Brazil: the first canine cases described in Ji-Paraná, Rondônia, Brazil / A disseminação da leishmaniose visceral no Brasil: os primeiros casos caninos descritos em Ji-Paraná, Rondônia, Brasil

Abstract Leishmaniases are zoonotic diseases caused by protozoa of the genus Leishmania. The disease has two clinical manifestations described in humans: visceral (VL) and cutaneous (CL) leishmaniasis. In Brazil, there has been an expansion of human VL. The participation of the dog as a reservoir of Leishmania infantum, the agent of VL, is important for the epidemiology of the disease since canine cases generally precede human cases. The present study aimed to evaluate the occurrence of Leishmania spp. infection in dogs in the municipality of Ji-Paraná by PCR assays using blood samples. Leishmania DNA was detected in two of the 105 studied dogs. The PCR products were sequenced and confirmed that the two samples (1.90%) correspond to L. infantum. The dogs had allochthonous history. Therefore, the positive results found here should serve as a warning to public health agencies. This is because Ji-Paraná is the third municipality to register cases of canine leishmaniasis (CanL) in Rondônia state. Thus, reinforcing the importance of expanding studies on the epidemiology and surveillance of VL in the region.

Resumo As leishmanioses são doenças causadas por protozoários do gênero Leishmania. A doença apresenta duas manifestações clínicas: leishmaniose visceral (LV) e cutânea (LC). No Brasil, a LV está em expansão. A participação do cão como reservatório é importante para a epidemiologia da doença, pois os casos caninos geralmente precedem os humanos. O presente estudo avaliou a ocorrência de LV em cães (LVC) do município de Ji-Paraná por meio de ensaios de PCR, utilizando-se amostras de sangue. O DNA de Leishmania foi detectado em dois dos 105 cães estudados. Os produtos da PCR foram sequenciados e confirmaram que duas amostras (1,90%) eram Leishmania infantum. Os cães tinham histórico alóctone. Os resultados positivos encontrados servem de alerta aos órgãos públicos de saúde. Isso porque Ji-Paraná é o terceiro município a registrar casos de LVC no estado de Rondônia. Dessa forma, reforça-se a importância da ampliação dos estudos sobre a epidemiologia e vigilância da LV na região.

Occurrence of Leishmania infection in the immediate geographic region of Ji-Paraná, Rondônia State, Brazil

Abstract INTRODUCTION: This study evaluated the epidemiology of American cutaneous leishmaniasis in the immediate region of Ji-Paraná, Rondônia State. METHODS: Samples and epidemiological data were collected from 105 patients. RESULTS: Leishmania infection was observed in 58 (55.2%) patients, and Leishmania braziliensis was present in 82.9% of the 41 sequenced samples. Infected patients were predominantly male (93.1%). Leishmania infection was twice as prevalent among rural inhabitants versus urban inhabitants. Lesions were more frequent in the upper limbs (arms/hands, 41.82%). CONCLUSIONS: The present data corroborate the zoonotic profile of cutaneous leishmaniasis; this information could help to improve surveillance and control strategies.

Entendimento dos estudantes de medicina a respeito da abrangência clínica da especialidade da cirurgia plástica / Understanding of medical students regarding the clinical scope of the plastic surgery specialty

Introdução: A cirurgia plástica é popularmente conhecida pela atuação na estética corporal. Além disso, engloba queimaduras, microcirurgia, cirurgia de mão e craniomaxilofacial. Devido à diversidade de especialidades médicas, muitos dos procedimentos em que os cirurgiões plásticos são capacitados, são também realizados por outros especialistas, o que pode justificar o desconhecimento da população em relação à rica atuação plástica. O objetivo é compreender o entendimento dos estudantes brasileiros de medicina sobre a abrangência clínica da cirurgia plástica. Métodos: Estudo transversal, com aplicação de questionário online pela plataforma Google Docs com critérios de inclusão ser estudante de medicina e possuir mais de dezoito anos. Composto por doze casos clínicos, enviado aos participantes por meio das redes sociais. Ao final de 99 dias com o link sendo divulgado, o questionário foi encerrado e as respostas contabilizadas com Windows Excel. Resultados: Amostra composta por 423 participantes, sendo maioria feminina e média de 22,23 anos, com maior prevalência do terceiro ano pertencente ao ciclo clínico acadêmico. Percebeu-se que os estudantes estão familiarizados com a ação dos cirurgiões plásticos no campo estético, reconstrução mamária após carcinoma e tratamento reparador de queimaduras. Já o tratamento de fraturas no complexo craniomaxilofacial e a abordagem da laceração dos tendões palmares foi pouco indicada pelos acadêmicos. Conclusão: Os estudantes apresentam um entendimento satisfatório da atuação dos cirurgiões plásticos no campo estético.

Introduction: Plastic surgery is popularly known for its performance in body aesthetics. Besides, it includes burns, microsurgery, hand and craniomaxillofacial surgery. Due to the diversity of medical specialties, many of the procedures in which plastic surgeons are trained are also performed by other specialists, which may justify the population's lack of knowledge regarding the rich plastic work. The objective is to understand the Brazilian medical students' comprehension of the plastic surgery clinical scope. Methods: Cross-sectional study, applying an online questionnaire with Google Docs platform using the inclusion criteria of being a medical student and over eighteen years old. It consisted of twelve clinical cases sent to participants through social networks. At the end of 99 days after the link's release, the questionnaire was closed, and the answers were counted with Windows Excel. Results: Sample composed of 423 participants, with a female majority and an average age of 22.23 years, with a higher prevalence of the third year of the academic clinical cycle. It was noticed that students are familiar with plastic surgeons' actions in the aesthetic field, breast reconstruction after carcinoma, and reparative treatment of burns. Academics poorly indicated the treatment of fractures in the craniomaxillofacial complex and the palmar tendons' laceration approach. Conclusion: Students have a good understanding of the role of plastic surgeons in the aesthetic field.

The CHEK2 Variant C.349A>G Is Associated with Prostate Cancer Risk and Carriers Share a Common Ancestor.

The identification of recurrent founder variants in cancer predisposing genes may have important implications for implementing cost-effective targeted genetic screening strategies. In this study, we evaluated the prevalence and relative risk of the CHEK2 recurrent variant c.349A>G in a series of 462 Portuguese patients with early-onset and/or familial/hereditary prostate cancer (PrCa), as well as in the large multicentre PRACTICAL case-control study comprising 55,162 prostate cancer cases and 36,147 controls. Additionally, we investigated the potential shared ancestry of the carriers by performing identity-by-descent, haplotype and age estimation analyses using high-density SNP data from 70 variant carriers belonging to 11 different populations included in the PRACTICAL consortium. The CHEK2 missense variant c.349A>G was found significantly associated with an increased risk for PrCa (OR 1.9; 95% CI: 1.1-3.2). A shared haplotype flanking the variant in all carriers was identified, strongly suggesting a common founder of European origin. Additionally, using two independent statistical algorithms, implemented by DMLE+2.3 and ESTIAGE, we were able to estimate the age of the variant between 2300 and 3125 years. By extending the haplotype analysis to 14 additional carrier families, a shared core haplotype was revealed among all carriers matching the conserved region previously identified in the high-density SNP analysis. These findings are consistent with CHEK2 c.349A>G being a founder variant associated with increased PrCa risk, suggesting its potential usefulness for cost-effective targeted genetic screening in PrCa families.

Hereditary Predisposition to Prostate Cancer: From Genetics to Clinical Implications.

Prostate cancer (PrCa) ranks among the top five cancers for both incidence and mortality worldwide. A significant proportion of PrCa susceptibility has been attributed to inherited predisposition, with 10-20% of cases expected to occur in a hereditary/familial context. Advances in DNA sequencing technologies have uncovered several moderate- to high-penetrance PrCa susceptibility genes, most of which have previously been related to known hereditary cancer syndromes, namely the hereditary breast and ovarian cancer (BRCA1, BRCA2, ATM, CHEK2, and PALB2) and Lynch syndrome (MLH1, MSH2, MSH6, and PMS2) genes. Additional candidate genes have also been suggested, but further evidence is needed to include them in routine genetic testing. Recommendations based on clinical features, family history, and ethnicity have been established for more cost-efficient genetic testing of patients and families who may be at an increased risk of developing PrCa. The identification of alterations in PrCa predisposing genes may help to inform screening strategies, as well as treatment options, in the metastatic setting. This review provides an overview of the genetic basis underlying hereditary predisposition to PrCa, the current genetic screening recommendations, and the implications for clinical management of the disease.

The effect of sample size on polygenic hazard models for prostate cancer.

We determined the effect of sample size on performance of polygenic hazard score (PHS) models in prostate cancer. Age and genotypes were obtained for 40,861 men from the PRACTICAL consortium. The dataset included 201,590 SNPs per subject, and was split into training and testing sets. Established-SNP models considered 65 SNPs that had been previously associated with prostate cancer. Discovery-SNP models used stepwise selection to identify new SNPs. The performance of each PHS model was calculated for random sizes of the training set. The performance of a representative Established-SNP model was estimated for random sizes of the testing set. Mean HR98/50 (hazard ratio of top 2% to average in test set) of the Established-SNP model increased from 1.73 [95% CI: 1.69-1.77] to 2.41 [2.40-2.43] when the number of training samples was increased from 1 thousand to 30 thousand. Corresponding HR98/50 of the Discovery-SNP model increased from 1.05 [0.93-1.18] to 2.19 [2.16-2.23]. HR98/50 of a representative Established-SNP model using testing set sample sizes of 0.6 thousand and 6 thousand observations were 1.78 [1.70-1.85] and 1.73 [1.71-1.76], respectively. We estimate that a study population of 20 thousand men is required to develop Discovery-SNP PHS models while 10 thousand men should be sufficient for Established-SNP models.

Direct defense elicited by tetranychus urticae koch (acari: tetranychidae) in bt maize plants / Defesa direta elicitada por Tetranychus urticae koch (Acari: Tetranychidae) em milho Bt

Direct and indirect induced defense mechanisms against herbivores can be manifested in maize (Zea mays L.) plants. Furthermore, there are constitutive defenses in which plants continuously express resistance traces. In recent decades has increased the production of transgenic maize plants that constitutively express proteins with insecticide action (Bt maize). The increase of the use of transgenic maize cultivars with the Bt (Bacillus thuringiensis) gene demand studies that evaluate the impacts caused by this technology on plant defense mechanisms and their impact on non-targeted organisms, as the two-spotted spider mite Tetranychus urticae Koch (Acari: Tetranychidae). We tested the hypothesis that Bt maize plants (expressing Cry1F protein) would be capable of inducing direct defenses to T. urticae after being attacked by these mites. Thus, we used plants of a commercial maize hybrid (30F35 Hx - expressing Cry1F protein) and plants of its respective non-Bt isogenic line (control). We compared the survival and reproductive performances of T. urticae on plants of both lines that were previously infested with conspecifics and on plants that did not suffer pre-infestations. The previous infestation of maize plants by T. urticae did not impacted the survival and reproductive abilities of adult and immature forms of the conspecific in both genotypes. These results suggest that, Bt maize expressing the Cry1F insecticidal protein, does not interfere in the induction of direct defense by the T. urticae when compared with conventional maize plants.

Mecanismos diretos e indiretos de defesa induzida contra herbívoros podem manifestar-se em plantas de milho (Zea mays.). Além das defesas induzidas, existem as defesas constitutivas, nas quais as plantas expressam a resistência de forma contínua. Nas últimas décadas vem se difundindo a produção deplantas de milho geneticamente modificadas que expressam proteínas com ação inseticida de forma constitutiva (milho Bt). Com o crescente uso de cultivares de milho transgênico com o gene Bt (Bacillus thuringiensis), há uma demanda por estudos que avaliem os impactos causados por essa tecnologia sobre os mecanismos de defesa das plantas e seu impacto sobre organismos não alvo, como o ácaro-rajado Tetranychus urticae Koch (Acari: Tetranychidae). Testou-se a hipótese de que plantas de milho Bt (expressando a proteína Cry 1F) seriam capazes de induzir defesas diretas a T. urticae após o ataque por esses ácaros. Assim, foram utilizadas plantas de milhos híbridos comerciais (30F35 Hx expressando a proteína Cry 1F) e seu respectivo isogênico não-Bt (controle). Nós comparamos a sobrevivência e o desempenho reprodutivo de T. urticae em plantas de ambas as linhagens que foram previamente infestadas com coespecíficos e em plantas que não foram pré-infestadas. A infestação prévia de plantas de milho Bt por T. urticae nãoapresentou diferença nos padrões de sobrevivência de formas adultas e formas jovens do coespecífico em comparação com o milho convencional. Os resultados sugerem que, o fato de o milho Bt expressar a toxina inseticida Cry 1F, não interfere na indução de defesa direta pelo ácaro-rajado T. urticae quando comparado com plantas de milho convencional.

Brazil's first free-mating laboratory colony of Nyssorhynchus darlingi

Abstract INTRODUCTION: The lack of highly-productive Nyssorhynchus darlingi laboratory colonies limits some studies. We report the first well-established laboratory colony of Ny. darlingi in Brazil. METHODS: Mosquitoes were collected from Porto Velho and were reared at the Laboratory of Fiocruz/RO. After induced mating by light stimulation in the F1 to F6, the subsequent generations were free mating. Larvae were reared in distilled water and fed daily until pupation. RESULTS: In 11 generations, the colony produced a high number of pupae after the F5 generation. CONCLUSIONS: These results demonstrate the potential for permanently establishing Ny. darlingi colonies for research purposes in Brazil.

Diversity, natural infection and blood meal sources of phlebotomine sandflies (Diptera, Psychodidae) in the western Brazilian Amazon

BACKGROUND The state of Rondônia (RO) is a hot spot for human cases of cutaneous leishmaniasis. Many sandfly species in RO are putative vectors of leishmaniasis. OBJECTIVES This study examines the diversity patterns and the presence of Leishmania DNA and blood meal sources of sandflies in RO. METHODS A sandfly survey was performed between 2016 and 2018 in 10 municipalities categorised into three different environment types: (i) Conservation Unit (CUN) - comprised of preserved ombrophilous forests; (ii) Forest Edge (FE) - small forest fragments; and (iii) Peridomicile (PE) - areas around dwellings. FINDINGS A total of 73 species were identified from 9,535 sandflies. The most abundant species were Psychodopygus davisi (1,741 individuals), Nyssomyia antunesi (1,397), Trichophoromyia auraensis (1,295) and Trichophoromyia ubiquitalis (1,043). Diversity was the highest in CUN, followed by the FE and PE environments. One pool of Ps. davisi tested positive for Leishmania braziliensis, reinforcing the possibility that Ps. davisi acts as a vector. The cytochrome b (cytb) sequences were used to identify three blood meal sources: Bos taurus, Homo sapiens and Tamandua tetradactyla. MAIN CONCLUSIONS Our results demonstrated that sandflies can switch between blood meal sources in differing environments. This study enhances the knowledge of the vector life cycle in RO and provides information relevant to leishmaniasis surveillance.

Targeted next generation sequencing identifies functionally deleterious germline mutations in novel genes in early-onset/familial prostate cancer.

Considering that mutations in known prostate cancer (PrCa) predisposition genes, including those responsible for hereditary breast/ovarian cancer and Lynch syndromes, explain less than 5% of early-onset/familial PrCa, we have sequenced 94 genes associated with cancer predisposition using next generation sequencing (NGS) in a series of 121 PrCa patients. We found monoallelic truncating/functionally deleterious mutations in seven genes, including ATM and CHEK2, which have previously been associated with PrCa predisposition, and five new candidate PrCa associated genes involved in cancer predisposing recessive disorders, namely RAD51C, FANCD2, FANCI, CEP57 and RECQL4. Furthermore, using in silico pathogenicity prediction of missense variants among 18 genes associated with breast/ovarian cancer and/or Lynch syndrome, followed by KASP genotyping in 710 healthy controls, we identified "likely pathogenic" missense variants in ATM, BRIP1, CHEK2 and TP53. In conclusion, this study has identified putative PrCa predisposing germline mutations in 14.9% of early-onset/familial PrCa patients. Further data will be necessary to confirm the genetic heterogeneity of inherited PrCa predisposition hinted in this study.

Desinfecção de água cinza por fotocatálise heterogênea / Disinfection of greywater by heterogeneous photocatalysis

RESUMO A desinfecção adequada de águas cinzas é necessária para garantir a segurança de sua reutilização, principalmente em aplicações com potencial de exposição humana. Diversos processos oxidativos avançados têm sido empregados nos últimos anos para a degradação de contaminantes orgânicos, bem como para desinfecção de águas e efluentes. O objetivo deste trabalho foi testar TiO2 suportado em microtubos para desinfetar águas cinzas por meio da fotocatálise heterogênea, visando ao reúso hídrico em bacia sanitária. A água cinza utilizada nos experimentos foi coletada após passar por um sistema de tratamento composto de um tanque de evapotranspiração seguido de banhado construído de fluxo horizontal. Foram realizados testes em batelada utilizando-se um reator fotoquímico cilíndrico de 1,0 L (volume total do reator), preenchido com pequenos cilindros de vidro do tipo Pyrex com TiO2 suportado. Para os testes de desinfecção, foram utilizados os processos UV, H2O2, UV/TiO2, UV/H2O2 e UV/TiO2/H2O2. Foi possível obter uma camada homogênea de TiO2 depositada nos pequenos tubos de vidro Pyrex, com espessura média de 35,3 µm, capaz de promover um incremento na desinfecção de águas cinzas. Porém, mesmo com um maior poder de desinfecção do TiO2 - se comparado com a fotólise (UV) -, os processos em que se empregou o peróxido de hidrogênio foram bem mais eficientes, tanto na desinfecção (inativação total de coliformes totais, enterococos e Pseudomonas aeruginosa) quanto na remoção de matéria orgânica em termos de demanda química de oxigênio (em torno de 60%). As amostras mantidas em temperatura ambiente e envolvidas por plástico escuro não apresentaram recrescimento bacteriano com 24h de armazenamento após os experimentos, mostrando assim a viabilidade da água cinza tratada em reúso doméstico.

ABSTRACT Proper disinfection of greywater is needed to ensure the safety of its reuse, especially in applications with potential for human exposure. Various advanced oxidation processes have been used in recent years for the degradation of organic contaminants, as well as for disinfection of water and wastewater. The purpose of this study was to test TiO2 supported in microtubes to disinfect greywater by photocatalysis in order to reuse it in sanitary bowl. The greywater used in the experiments was collected after passing through a treatment system consisting of an evapotranspiration tank followed by constructed wetland with horizontal flow. Batch tests were conducted using a cylindrical photochemical reactor of 1.0 L (total volume of the reactor), filled with small glass Pyrex cylinders with supported TiO2. For disinfection tests, the processes UV, H2O2, UV/TiO2, UV/H2O2 and UV/TiO2/H2O2 were used. It was possible to obtain a homogeneous layer of TiO2 deposited in small Pyrex glass tubes with an average thickness of 35.3 µm; this layer was able to promote an increase in the greywater disinfection. However, even with a greater disinfection power of TiO2 compared with photolysis (UV), the processes with hydrogen peroxide was much more efficient in disinfection (total inactivation of total coliforms, enterococci and Pseudomonas aeruginosa) and in the removal of organic matter in terms of chemical oxygen demand (around 60%). Samples stored at a room temperature and wrapped in plastic dark showed no bacterial regrowth after 24 hours of storage after the experiments, thus showing the viability of treated greywater for domestic reuse.

Validation of a Next-Generation Sequencing Pipeline for the Molecular Diagnosis of Multiple Inherited Cancer Predisposing Syndromes.

Despite the growing knowledge of the genetic background behind the cancers that occur in a context of hereditary predisposition, personal or family cancer history may not be clear enough to support directional gene testing. Defined targeted next-generation sequencing gene panels allow identification of the causative disease mutations of multigene syndromes and differential diagnosis for syndromes with phenotypically overlapping characteristics. Herein, we established a next-generation sequencing analysis pipeline for the molecular diagnosis of multiple inherited cancer predisposing syndromes using the commercially available target sequencing panel TruSight Cancer. To establish the analysis pipeline, we included 22 control samples with deleterious mutations covering all genes currently analyzed at our institution by standard Sanger sequencing. We tested the pipeline using 51 samples from patients with a clinical diagnosis of neurofibromatosis type 1 (NF1), 10 of which without previous molecular characterization of the causative NF1 mutations. We propose a thoroughly validated analysis pipeline that combines Isaac Enrichment, Burrows-Wheeler Aligner Enrichment, and NextGENe for the alignment and variant calling, and GeneticistAssistant for variant annotation and prioritization. This pipeline allowed the identification of disease-causing mutations in all 73 patients, including a large duplication of 37 bp in NF1. We show that high sensitivity and specificity can be achieved by using multiple bioinformatic tools for alignment and variant calling and careful variant filtering, having in mind the clinical question.

Factors associated with suicidal ideation among university students.

Objective: to analyze the factors associated with suicidal ideation in a representative sample of university students. Methods: cross-sectional study, carried out with 637 students of the Federal University of Mato Grosso. The presence of suicidal ideation, demographic and socioeconomic variables, use of alcohol through the Alcohol, Smoking and Substance Involvement Screening Test, and depressive symptoms (Major Depression Inventory) were investigated. Bivariate analysis was performed with the Chi-square test and multivariate analysis using the Poisson regression model. Results: it was found that 9.9% of the students had suicidal thoughts in the previous 30 days and, in the bivariate analysis, the variables economic class, sexual orientation, religious practice, suicide attempts in the family and among friends, alcohol consumption and depressive symptoms were associated with suicidal ideation. In the multivariate analysis sexual orientation, suicide attempts in the family and the presence of depressive symptoms remained as associated factors. Conclusion: these findings constitute a situational diagnosis that enables the formulation of academic policies and preventive actions to confront this situation on the university campus.

OBJETIVO: analisar os fatores associados à ideação suicida em uma amostra representativa de estudantes universitários. MÉTODOS: Revisão sistemática da literatura. A busca foi realizada nas bases de dados PubMed, Lilacs e Web of Science, sem restrição de datas e idiomas, entretanto foram incluídos somente os artigos publicados em português, inglês e espanhol. Foram critérios de inclusão: ter delineamento observacional; possuir os fatores socioeconômicos como variáveis de interesse na análise do acesso ou utilização de serviços de saúde entre idosos; ter amostra representativa da população alvo; fazer ajuste para fatores de confusão; e não apresentar viés de seleção. OBJETIVOS: Foram encontrados 5.096 artigos após a exclusão de duplicidades e 36 foram selecionados para a revisão após o processo de leitura e avaliação dos critérios de inclusão. Maior renda e escolaridade estiveram associadas à utilização e acesso a consultas médicas nos países em desenvolvimento e em alguns países desenvolvidos. A mesma associação foi observada nas consultas odontológicas em todos os países. A maioria dos estudos não apresentou associação entre características socioeconômicas e uso de serviços de internação e emergência. Foi identificado maior uso de visita domiciliar em indivíduos de menor renda, com exceção dos Estados Unidos. CONCLUSÕES: Observou-se desigualdade no acesso ou na utilização de serviços de saúde na maior parte dos países, variando em relação ao tipo de serviço utilizado. A ampliação da cobertura de serviços de saúde faz-se necessária para a redução da desigualdade no acesso gerada por iniquidades sociais.