Epigenetic control of adamantinomatous craniopharyngiomas.

INTRODUCTION: Studies addressing the methylation pattern in adamantinomatous craniopharyngioma (ACP) are lacking. OBJECTIVE: To identify methylation signatures in ACPs regarding clinical presentation and outcome. METHODS: Clinical and pathology data were collected from 35 ACP patients (54% male; 18.1 years [2-68]). CTNNB1 mutations and methylation profile (MethylationEPIC/Array-Illumina) were analyzed in tumoral DNA. Unsupervised machine learning analysis of this comprehensive methylome sample was achieved using hierarchical clustering and multi-dimensional scaling. Statistical associations between clusters and clinical features were achieved using Fisher's test and global biological process interpretations were aided by Gene Ontology enrichment analyses. RESULTS: Two clusters were revealed consistently by all unsupervised methods (ACP-1: n = 18; ACP-2: n = 17) with strong bootstrap statistical support. ACP-2 was enriched by CTNNB1 mutations (100% vs 56%, P = 0.0006), hypomethylated in CpG Island (CGI), non-CGI sites, and globally (P < 0.001), and associated with greater tumor size (24.1 vs 9.5cm3, P = 0.04). Enrichment analysis highlighted pathways on signaling transduction, transmembrane receptor, development of anatomical structures, cell-adhesion, cytoskeleton organization, and cytokine binding, and also cell-type specific biological processes as regulation of oligodendrocytes, keratinocyte, and epithelial cells differentiation. CONCLUSION: Two clusters of ACP patients were consistently revealed by unsupervised machine learning methods, being one of them significantly hypomethylated, enriched by CTNNB1 mutated ACPs, and associated with increased tumor size. Enrichment analysis reinforced pathways involved in tumor proliferation and in cell-specific tumoral microenvironment.

Glenoid Bone Loss - A Retrospective Evaluation of Functional Outcomes after Bone Block Surgery for Anterior Shoulder Instability in High-demand Athletes.

Objective This study assesses the relationship between the glenoid bone loss size and range of motion, functional outcomes, and complications in high-performance athletes undergoing bone block surgery for anterior shoulder instability. Methods This retrospective study evaluated postoperative outcomes in athletes submitted to bone block surgery for anterior shoulder instability. In 5 years, 41 shoulders underwent the procedure; 20 had bone losses up to 15%, and 21 shoulders presented bone losses ranging from 15% and 25%. Results There was no statistically significant difference regarding postoperative complications, new dislocations, and the rate of return to sports. In addition, the quantitative criteria evaluated, i.e., ranges of motion and functional scores, showed no statistically significant difference between groups. Conclusion The size of the bone loss per se does not seem to affect functional outcomes and complications from these procedures, which are safe techniques for small and large bone losses.

Profound N-glycan remodelling accompanies MHC-II immunopeptide presentation.

Immunopeptidomics, the study of peptide antigens presented on the cell surface by the major histocompatibility complex (MHC), offers insights into how our immune system recognises self/non-self in health and disease. We recently discovered that hyper-processed (remodelled) N-glycans are dominant features decorating viral spike immunopeptides presented via MHC-class II (MHC-II) molecules by dendritic cells pulsed with SARS-CoV-2 spike protein, but it remains unknown if endogenous immunopeptides also undergo N-glycan remodelling. Taking a multi-omics approach, we here interrogate published MHC-II immunopeptidomics datasets of cultured monocyte-like (THP-1) and breast cancer-derived (MDA-MB-231) cell lines for overlooked N-glycosylated peptide antigens, which we compare to their source proteins in the cellular glycoproteome using proteomics and N-glycomics data from matching cell lines. Hyper-processed chitobiose core and paucimannosidic N-glycans alongside under-processed oligomannosidic N-glycans were found to prevalently modify MHC-II-bound immunopeptides isolated from both THP-1 and MDA-MB-231, while complex/hybrid-type N-glycans were (near-)absent in the immunopeptidome as supported further by new N-glycomics data generated from isolated MHC-II-bound peptides derived from MDA-MB-231 cells. Contrastingly, the cellular proteomics and N-glycomics data from both cell lines revealed conventional N-glycosylation rich in complex/hybrid-type N-glycans, which, together with the identification of key lysosomal glycosidases, suggest that MHC-II peptide antigen processing is accompanied by extensive N-glycan trimming. N-glycan remodelling appeared particularly dramatic for cell surface-located glycoproteins while less remodelling was observed for lysosomal-resident glycoproteins. Collectively, our findings indicate that both under- and hyper-processed N-glycans are prevalent features of endogenous MHC-II immunopeptides, an observation that demands further investigation to enable a better molecular-level understanding of immune surveillance.

Impact of Compression on the Textural and Structural Properties of CPO-27(Ni).

The employment of metal-organic frameworks in powder form is undesirable from an industrial perspective due to process and safety issues. This work is devoted to evaluating the impact of compression on the textural and structural properties of CPO-27(Ni). For this purpose, CPO-27(Ni) was synthesized under hydrosolvothermal conditions and characterized. Then, the resulting powder was compressed into binderless pellets using variable compression forces ranging from 5-90 kN (37-678 MPa) and characterized by means of nitrogen adsorption/desorption, thermogravimetric analysis and powder X-ray diffraction to evaluate textural, thermal and structural changes. Both textural and structural properties decreased with increasing compression force. Thermal stability was impacted in pellets compressed at forces over 70 kN. CPO-27(Ni) pelletized at 5, 8 and 10 kN, and retained more than 94% of its initial textural properties, while a loss of about one-third of the textural property was observed for the two most compressed samples (70 and 90 kN) compared to the starting powder.

EVALUATION OF THE EPIDEMIOLOGY OF EXPOSED FRACTURES BEFORE AND DURING THE COVID-19 PANDEMIC.

Objective: To assess the impact of the COVID-19 pandemic on the epidemiology and clinical outcomes of open fractures considering the periods before and during the pandemic. Methods: An observational and retrospective study, which included patients aged over 18 years, admitted to the Orthopedics and Traumatology Ward of Hospital São Paulo, of the Federal University of São Paulo (UNIFESP). Data was collected in two moments: pre-pandemic (March 1, 2019, to February 29, 2020) and during the pandemic (March 1, 2020, to February 28, 2021). Results: In total, 183 patients were evaluated with a mean age of 36 years ± 14 years. In the pre-pandemic period, 94 patients underwent surgery, 81 men (85.37%) and 13 women (14.2%), with a mean age of 36 ± 3 years. During the pandemic period, 89 patients were subjected to surgery, 77 men (86.6%) and 12 women (13.4%), with a mean age of 38 ± 3 years. Conclusion: During the pandemic, open fractures were still more common in men. Regarding hospital indicators, the prevalence of infections in the surgical wound and the length of stay of patients with open fractures increased, however, with little significance. Fractures classified as Gustilo IIIA were the most common, while the most common according to the AO classification were 33, 34, 42, 43, 2R3, and 2R3 + 2U2. The frequency of run overs during the pandemic decreased. However, firearm projectile injuries and falls and occupational injuries increased. Level of Evidence III, Retrospective Comparative Study.

Objetivo: Avaliar o impacto da COVID-19 na epidemiologia e nos desfechos clínicos das fraturas expostas considerando os períodos antes e durante a pandemia. Métodos: Estudo observacional e retrospectivo que incluiu pacientes maiores de 18 anos, internados na Enfermaria de Ortopedia e Traumatologia do Hospital São Paulo, da Universidade Federal de São Paulo. Os dados foram coletados em dois momentos - antes (março de 2019 a fevereiro de 2020) e durante a pandemia (março de 2020 a fevereiro de 2021) -, por meio da análise de prontuários eletrônicos de todos os registros hospitalares dessa instituição. Resultados: No total, foram avaliados 183 pacientes com média de idade de 36 ± 14 anos. No período pré-pandêmico, foram operados 94 pacientes, sendo 81 homens (85,37%) e 13 mulheres (14,2%), com média de idade de 36 ± 3 anos. Já ao longo do período pandêmico, foram operados 89 pacientes, sendo 77 homens (86,6%) e 12 mulheres (13,4%), com média de idade de 38 ± 3 anos. Conclusão: Durante a pandemia, a ocorrência de fraturas expostas se manteve com maior frequência em indivíduos do sexo masculino. Quanto aos indicadores hospitalares, houve aumento da prevalência de infecções na ferida operatória, assim como do tempo de internação dos pacientes, todavia, com pouca significância. Foram mais frequentes as fraturas classificadas como Gustilo IIIA e, pela classificação AO, as fraturas 33, 34, 42, 43, 2R3 e 2R3 + 2U2. Notamos redução da frequência de atropelamentos durante a pandemia e aumento dos casos de ferimentos por projétil de arma de fogo, quedas e acidentes ocupacionais. Nível de Evidência III, Estudo Retrospectivo Comparativo.

GATA-3 expression and its correlation with prognostic factors and survival in canine mammary tumors.

Introduction: The transcription factor GATA-3 plays a significant role in mammary gland development and differentiation. Recent studies on human oncology have demonstrated its association with favorable pathologic factors in breast cancer. Canine mammary tumours, proposed as comparative and translational study models, have epidemiological, clinical, biological, and genetic characteristics similar to those of human breast cancers. Methods: Here, we evaluated the frequency of GATA-3 expression in mammary tumors of dogs and its relationship with prognostic factors and survival. Tumor samples were obtained from 40 female dogs and grouped according to histological type into benign tumors (n = 10), carcinoma in mixed tumors (CMTs) (n = 20), and aggressive tumors (n = 10). CMTs were further separated according to histological grade, and data on clinical staging and diagnosis, histopathological grading, and survival rate were collected. Results: GATA-3 and estrogen receptor (ER) expression were higher in benign and well-differentiated carcinomas than in aggressive tumors, which showed greater Ki-67 expression. The expression rate of ER in the studied groups was equivalent to that of GATA-3. We identified a strong positive correlation between GATA-3 and ER expression frequencies and a negative correlation between those of GATA-3 and Ki-67. There were associations between GATA-3 (p < 0.001), Ki-67 (p = 0.003), tumor size (p < 0.001), clinical stage (p = 0.002), lymph node metastasis (p < 0.001), and histological grade (p < 0.001) by univariate survival analysis. The parameters ER (p = 0.015) and GATA-3 (p = 0.005) also influenced survival in a multifactorial manner. Discussion: Kaplan-Meier analysis of survival curves validated our previous findings that dogs with GATA-3 expression in ≥79.4% of cells had significantly higher survival rates (p < 0.001). The performance analysis showed that the expression of GATA-3 in ≥79.4% of cells effectively predicted survival or death in dogs with mammary tumors. Collectively, these results suggest that GATA-3 can be a relevant marker in the study of mammary tumor progression and has potential as a prognosis marker for predicting outcomes in canine mammary tumors.

SAPrIm, a semi-automated protocol for mid-throughput immunopeptidomics.

Human leukocyte antigen (HLA) molecules play a crucial role in directing adaptive immune responses based on the nature of their peptide ligands, collectively coined the immunopeptidome. As such, the study of HLA molecules has been of major interest in the development of cancer immunotherapies such as vaccines and T-cell therapies. Hence, a comprehensive understanding and profiling of the immunopeptidome is required to foster the growth of these personalised solutions. We herein describe SAPrIm, an Immunopeptidomics tool for the Mid-Throughput era. This is a semi-automated workflow involving the KingFisher platform to isolate immunopeptidomes using anti-HLA antibodies coupled to a hyper-porous magnetic protein A microbead, a variable window data independent acquisition (DIA) method and the ability to run up to 12 samples in parallel. Using this workflow, we were able to concordantly identify and quantify ~400 - 13000 unique peptides from 5e5 - 5e7 cells, respectively. Overall, we propose that the application of this workflow will be crucial for the future of immunopeptidome profiling, especially for mid-size cohorts and comparative immunopeptidomics studies.

Cerebral vasospasm following subarachnoid hemorrhage: a rare complication after transsphenoidal surgery for pituitary macroadenoma.

Summary: Postoperative (PO) complications after transsphenoidal surgery (TSS) are rare when performed in pituitary referral centers. Partial hypopituitarism is more frequent and somewhat expected. Meningitis, cerebrospinal fluid leaks, and visual deficits are unusual. Cerebrovascular complications, including cerebral vasospasm are rare, usually under-appreciated and not mentioned to the patient prior to the surgery. This is a report of a 51-year-old male with a non-functioning pituitary macroadenoma presenting with partial hypopituitarism and visual field loss. The patient was submitted to an uneventful TSS. On the first PO day, he developed a left palpebral ptosis with unequal pupils and impaired consciousness (12 points on Glasgow Coma Scale). CT scan revealed a perimesencephalic subarachnoid hemorrhage (SAH) grade 1 according to the modified Fisher scale. High-dose dexamethasone (16 mg/day) was initiated and the patient became more alert (Glasgow 14). On the fifth PO day, due to progression of the neurological deficits (left III, IV, and VI cranial nerves palsy, ataxia, dysdiadochokinesia, right dysmetria, and dysarthria), a magnetic resonance angiography was obtained and revealed a recent mesencephalic infarct without evident vasospasm. Nevertheless, nimodipine 60 mg 4/4 h was initiated. No improvement was seen after 3 days of treatment. The patient was discharged and put on rehabilitation, returning to normal gait and balance after 7 months. This, therefore, is a case of an unexpected mesencephalic infarct probably due to vasospasm induced by minor SAH. Although exceptionally rare, informing the patient about this event prior to TSS is important due to its significant neurological impact. More data are needed considering preventive treatment with nimodipine as soon as SAH is detected after TSS and whether it would improve neurological outcomes. Learning points: Whenever neurological deficits arise after transsphenoidal surgery (TSS), systemic infection, meningitis, electrolyte imbalance, and evident hemorrhage must be promptly investigated. Although rare, cerebral vasospasm (CVS) after TSS is associated with high morbidity and high mortality rates. Vigilance for vasospasm is necessary for patients undergoing TSS for pituitary adenoma, especially those with significant suprasellar extension. Informing this event to the patient prior to TSS is essential due to its significant morbidity and mortality. Post-TSS subarachnoid hemorrhage and hemiparesis may be important clues indicating CVS and infarction. There is limited evidence in the literature regarding post-TSS CVS surveillance and treatment strategies which could have an impact on clinical decisions.

Translation, adaptation, and evidence of content validity and internal structure of the brazilian version of the contest orientation scale / Tradução, adaptação e evidências de validade de conteúdo e de estrutura interna da versão brasileira da contest orientation scale

ABSTRACT This study aimed to translate, adapt, and present the first evidence of content validity and internal structure of the Brazilian version of the Contest Orientation Scale. To this end, three studies were conducted. The first study was the translation of the items of the scale and adaptation to the Brazilian sports context, considering cultural and theoretical aspects by four judges, fluent in both languages (English and Portuguese). The second study was the content validation of the translated version. To this end, three judges evaluated Language Clarity (CVCtLC= 0.90), Theoretical Relevance (CVCtTR= 0.91), and Theoretical Dimensionality (KTotal= 0.71). The results ranged from good to satisfactory. The third study applied the scale to 188 athletes of both sexes, with a mean age of 30.6 years. A semi-confirmatory factor analysis was performed, which presented a clean factorial solution, with adjustment ranging from reasonable to satisfactory and satisfactory reliability indices (α = 0.744). In conclusion, the Brazilian version of the scale presents indices favorable to its application to evaluate the orientations to the dispute in the sport.

RESUMO O objetivo do estudo foi traduzir, adaptar e apresentar as primeiras evidências de validade de conteúdo e de estrutura interna da versão brasileira da Contest Orientation Scale. Para tanto, foram realizados três estudos. No primeiro estudo, a tradução dos itens da escala e adaptação para o contexto esportivo brasileiro, considerando aspectos culturais e teóricos, foi realizada por quatro juízes, fluentes em ambos os idiomas (inglês e português). No segundo estudo, foi realizada a validação de conteúdo da versão traduzida. Para tanto, três juízes realizaram a avaliação da Clareza de Linguagem (CVCtCL = 0,90), Relevância Teórica (CVCtRT = 0,91) e Dimensionalidade Teórica (KTotal = 0,71). Os resultados variaram de bons a satisfatórios. No terceiro estudo, a escala foi aplicada a 188 atletas de ambos os sexos, com idade média de 30,6 anos. Realizou-se uma análise fatorial semi-confirmatória, que apresentou uma solução fatorial limpa, com ajuste variando de razoável a satisfatório e índices de fidedignidade satisfatórios (α = 0,744). Conclui-se que a versão brasileira da escala apresenta índices favoráveis à sua aplicação na avaliação das orientações à disputa no esporte.

Glenoid Bone Loss - A Retrospective Evaluation of Functional Outcomes after Bone Block Surgery for Anterior Shoulder Instability in High-demand Athletes / Falha óssea na glenoide - Uma avaliação retrospectiva dos desfechos funcionais após cirurgia de bloqueio ósseo para instabilidade anterior do ombro em atletas de alta demanda

Abstract Objective This study assesses the relationship between the glenoid bone loss size and range of motion, functional outcomes, and complications in high-performance athletes undergoing bone block surgery for anterior shoulder instability. Methods This retrospective study evaluated postoperative outcomes in athletes submitted to bone block surgery for anterior shoulder instability. In 5 years, 41 shoulders underwent the procedure; 20 had bone losses up to 15%, and 21 shoulders presented bone losses ranging from 15% and 25%. Results There was no statistically significant difference regarding postoperative complications, new dislocations, and the rate of return to sports. In addition, the quantitative criteria evaluated, i.e., ranges of motion and functional scores, showed no statistically significant difference between groups. Conclusion The size of the bone loss per se does not seem to affect functional outcomes and complications from these procedures, which are safe techniques for small and large bone losses.

Resumo Objetivo Avaliar a relação do tamanho do defeito ósseo da glenoide no arco de movimento, nos resultados funcionais e nas complicações em pacientes atletas de alta performance submetidos a cirurgia de bloqueio ósseo para instabilidade anterior do ombro. Método Estudo retrospectivo no qual foram avaliados os resultados pós-operatórios de atletas submetidos a cirurgia de bloqueio ósseo para instabilidade anterior do ombro. Em 5 anos foram 41 ombros operados, sendo 20 deles com até 15% de defeito ósseo e 21 com defeitos entre 15% e 25%. Resultados Não houve diferença estatisticamente significativa com relação a complicações pós-operatórias, novas luxações, e na taxa de retorno ao esporte. Os critérios quantitativos avaliados - arcos de movimento e escores funcionais - também não apresentaram diferença estatisticamente significativa entre os grupos. Conclusão O tamanho do defeito ósseo por si só não parece afetar os resultados funcionais e as complicações desses procedimentos, sendo uma técnica segura tanto para defeitos pequenos, quanto para os maiores.

Validity evidences of two Sports-Pedagogy-related scales: socio-educational and autonomy development in youth sports / Evidências de validade de duas escalas relacionadas à Pedagogia do Esporte: desenvolvimento socioeducativo e de autonomia no esporte infantojuvenil

Abstract Competition is the essence of sports and, when conceived having participants as references, it can contributes to the development of different pedagogical contents, among them, socio-educative aspects and autonomy. The purpose of this research was to demonstrate the first validity evidences of two scales for supporting socio-educational and autonomy development in youth sports; both are part of the Battery of Tests Gonçalves-Balbinotti of Pedagogical Contents' Development Support in Youth Sports. We aim to estimate their internal structures, test their stabilities and estimate their internal consistency. A sample of 210 coaches answered the scales related to socio-educational and autonomy development, which presented second order two-factor structures with significant factor loadings (> 0.40) and explaining 80.6% and 73.2% of the constructs' total variance, respectively. The results related to the model fit were satisfactory (X2/df < 2.00; AGFI > 0.95; RMSEA < 0.05; CFI > 0.95; TLI > 0.95). The results regarding the internal consistency (0.786 < α < 0.913 for the factors; αSE = 0.889; αAu = 0.870) assure the precision of the measures and the reliability of their uses according to their purposes. The results answer the main and specific purposes of this research and indicate the safe use of these two scales.

Resumo A competição é a essência do esporte e, quando concebida tendo como referência o praticante, pode contribuir para o desenvolvimento de diferentes conteúdos pedagógicos, dentre eles, aspectos socioeducativos e autonomia. O objetivo deste estudo foi demonstrar as primeiras evidências de validade de duas escalas de favorecimento ao desenvolvimento socioeducativo e de autonomia no esporte infantojuvenil; ambas compondo a Bateria de Testes Gonçalves-Balbinotti de Favorecimento ao Desenvolvimento de Conteúdos Pedagógicos no Esporte Infantojuvenil. Objetiva-se estimar as estruturas internas, testar suas estabilidades e estimar suas consistências internas. Uma amostra de 210 treinadores respondeu as escalas relacionadas ao desenvolvimento socioeducativo e de autonomia, as quais apresentaram estruturas de segunda ordem com saturações fatoriais significativas (> 0,40) e explicando 80,6% e 73,2% da variância total dos construtos, respectivamente. Os resultados relacionados ao ajuste do modelo foram satisfatórios (X2/df < 2,00; AGFI > 0,95; RMSEA < 0,05; CFI > 0,95; TLI > 0,95). Os resultados relativos à consistência interna (0,786 < α < 0,913 para os fatores; αSE = 0,889; αAu = 0,870) asseguram a precisão das medidas e a fidedignidade dos seus usos de acordo com o proposto. Os resultados respondem aos objetivos geral e específicos da pesquisa e indicam o uso seguro destas duas escalas.

Evaluation of the epidemiology of exposed fractures before and during the covid-19 pandemic / Avaliação da epidemiologia das fraturas expostas antes e durante a pandemia de covid-19

ABSTRACT Objective: To assess the impact of the COVID-19 pandemic on the epidemiology and clinical outcomes of open fractures considering the periods before and during the pandemic. Methods: An observational and retrospective study, which included patients aged over 18 years, admitted to the Orthopedics and Traumatology Ward of Hospital São Paulo, of the Federal University of São Paulo (UNIFESP). Data was collected in two moments: pre-pandemic (March 1, 2019, to February 29, 2020) and during the pandemic (March 1, 2020, to February 28, 2021). Results: In total, 183 patients were evaluated with a mean age of 36 years ± 14 years. In the pre-pandemic period, 94 patients underwent surgery, 81 men (85.37%) and 13 women (14.2%), with a mean age of 36 ± 3 years. During the pandemic period, 89 patients were subjected to surgery, 77 men (86.6%) and 12 women (13.4%), with a mean age of 38 ± 3 years. Conclusion: During the pandemic, open fractures were still more common in men. Regarding hospital indicators, the prevalence of infections in the surgical wound and the length of stay of patients with open fractures increased, however, with little significance. Fractures classified as Gustilo IIIA were the most common, while the most common according to the AO classification were 33, 34, 42, 43, 2R3, and 2R3 + 2U2. The frequency of run overs during the pandemic decreased. However, firearm projectile injuries and falls and occupational injuries increased. Level of Evidence III, Retrospective Comparative Study.

RESUMO Objetivo: Avaliar o impacto da COVID-19 na epidemiologia e nos desfechos clínicos das fraturas expostas considerando os períodos antes e durante a pandemia. Métodos: Estudo observacional e retrospectivo que incluiu pacientes maiores de 18 anos, internados na Enfermaria de Ortopedia e Traumatologia do Hospital São Paulo, da Universidade Federal de São Paulo. Os dados foram coletados em dois momentos - antes (março de 2019 a fevereiro de 2020) e durante a pandemia (março de 2020 a fevereiro de 2021) -, por meio da análise de prontuários eletrônicos de todos os registros hospitalares dessa instituição. Resultados: No total, foram avaliados 183 pacientes com média de idade de 36 ± 14 anos. No período pré-pandêmico, foram operados 94 pacientes, sendo 81 homens (85,37%) e 13 mulheres (14,2%), com média de idade de 36 ± 3 anos. Já ao longo do período pandêmico, foram operados 89 pacientes, sendo 77 homens (86,6%) e 12 mulheres (13,4%), com média de idade de 38 ± 3 anos. Conclusão: Durante a pandemia, a ocorrência de fraturas expostas se manteve com maior frequência em indivíduos do sexo masculino. Quanto aos indicadores hospitalares, houve aumento da prevalência de infecções na ferida operatória, assim como do tempo de internação dos pacientes, todavia, com pouca significância. Foram mais frequentes as fraturas classificadas como Gustilo IIIA e, pela classificação AO, as fraturas 33, 34, 42, 43, 2R3 e 2R3 + 2U2. Notamos redução da frequência de atropelamentos durante a pandemia e aumento dos casos de ferimentos por projétil de arma de fogo, quedas e acidentes ocupacionais. Nível de Evidência III, Estudo Retrospectivo Comparativo.

Myracrodruon urundeuva leaf lectin damages exochorionic cells and binds to the serosal cuticle of Aedes aegypti eggs.

In recent years, lectins have been identified as alternative agents against Aedes aegypti during the aquatic phases of its life cycle. For example, chitin-binding lectin from Myracrodruon urundeuva leaf (MuLL) can function as a larvicide. In this study, we investigated whether MuLL can also act as an ovicide against this insect. Aedes aegypti eggs were incubated with MuLL for 72 h to determine the concentration at which the hatching rate reduces by 50% (EC50). The effects of MuLL on the egg surface structure were evaluated using scanning electron microscopy (SEM), and the possible interaction of MuLL with the internal structures of eggs and embryos was investigated using MuLL-fluorescein isothiocyanate (FITC) conjugate. MuLL acted as an ovicidal agent with an EC50 of 0.88 mg/mL. The SEM analysis revealed that eggs treated with MuLL for 24 and 48 h no longer had tubercles and did not show a well-defined exochorionic network. In addition, deformation and degeneration of the surface were observed after 72 h. Fluorescence microscopy showed that MuLL penetrated the eggs 48 h after incubation and was detected in the upper portion of the embryo's gut. After 72 h, MuLL was observed in the serosal cuticle and digestive tract. In conclusion, MuLL can function as an ovicidal agent against A. aegypti through damage to the surface and internal structures of the eggs.

Revealing the differential protein profiles behind the nitrogen use efficiency in popcorn (Zea mays var. everta).

We investigated the proteomic profiles of two popcorn inbred lines, P2 (N-efficient and N-responsive) and L80 (N-inefficient and nonresponsive to N), under low (10% of N supply) and high (100% of N supply) nitrogen environments, associated with agronomic- and physiological-related traits to NUE. The comparative proteomic analysis allowed the identification of 79 differentially accumulated proteins (DAPs) in the comparison of high/low N for P2 and 96 DAPs in the comparison of high/low N for L80. The NUE and N uptake efficiency (NUpE) presented high means in P2 in comparison to L80 at both N levels, but the NUE, NUpE, and N utilization efficiency (NUtE) rates decreased in P2 under a high N supply. DAPs involved in energy and carbohydrate metabolism suggested that N regulates enzymes of alternative pathways to adapt to energy shortages and that fructose-bisphosphate aldolase may act as one of the key primary nitrate responsive proteins in P2. Proteins related to ascorbate biosynthesis and nitrogen metabolism increased their regulation in P2, and the interaction of L-ascorbate peroxidase and Fd-NiR may play an important role in the NUE trait. Taken together, our results provide new insights into the proteomic changes taking place in contrasting inbred lines, providing useful information on the genetic improvement of NUE in popcorn.

ggVolcanoR: A Shiny app for customizable visualization of differential expression datasets.

Volcano and other analytical plots (e.g., correlation plots, upset plots, and heatmaps) serve as important data visualization methods for transcriptomic and proteomic analyses. Customizable generation of these plots is fundamentally important for a better understanding of dysregulated expression data and is therefore instrumental for the ensuing pathway analysis and biomarker identification. Here, we present an R-based Shiny application, termed ggVolcanoR, to allow for customizable generation and visualization of volcano plots, correlation plots, upset plots, and heatmaps for differential expression datasets, via a user-friendly interactive interface in both local executable version and web-based application without requiring programming expertise. Compared to currently existing packages, ggVolcanoR offers more practical options to optimize the generation of publication-quality volcano and other analytical plots for analyzing and comparing dysregulated genes/proteins across multiple differential expression datasets. In addition, ggVolcanoR provides an option to download the customized list of the filtered dysregulated expression data, which can be directly used as input for downstream pathway analysis. The source code of ggVolcanoR is available at https://github.com/KerryAM-R/ggVolcanoR and the webserver of ggVolcanoR 1.0 has been deployed and is freely available for academic purposes at https://ggvolcanor.erc.monash.edu/.

Supporting Physiological Trait for Indirect Selection for Grain Yield in Drought-Stressed Popcorn.

The identification of traits associated with drought tolerance in popcorn is a contribution to support selection of superior plants under soil water deficit. The objective of this study was to choose morphological traits and the leaf greenness index, measured on different dates, to estimate grain yield (GY) and popping expansion (PE), evaluated in a set of 20 popcorn lines with different genealogies, estimated by multiple regression models. The variables were divided into three groups: morpho-agronomic traits-100-grain weight (GW), prolificacy (PR), tassel length (TL), number of tassel branches, anthesis-silking interval, leaf angle (FA) and leaf rolling (FB); variables related to the intensity of leaf greenness during the grain-filling period, at the leaf level, measured by a portable chlorophyll meter (SPAD) and at the canopy level, calculated as the normalized difference vegetation index (NDVI). The inbred lines were cultivated under two water conditions: well-watered (WW), maintained at field capacity, and water stress (WS), for which irrigation was stopped before male flowering. The traits GY (55%) and PE (28%) were most affected by water restriction. Among the morpho-agronomic traits, GW and PR were markedly reduced (>10%). Under dry conditions, the FA in relation to the plant stalk tended to be wider, the FB curvature greater and leaf senescence accelerated (>15% at 22 days after male flowering). The use of multiple regression for the selection of predictive traits proved to be a useful tool for the identification of groups of adequate traits to efficiently predict the economically most important features of popcorn (GY and PE). The SPAD index measured 17 days after male flowering proved useful to select indirectly for GY, while, among the morphological traits, TL stood out for the same purpose. Of all traits, PR was most strongly related with PE under WS, indicating its use in breeding programs. The exploitation of these traits by indirect selection is expected to induce increments in GY and PE.

Phenotyping Latin American Open-Pollinated Varieties of Popcorn for Environments with Low Water Availability.

Drought is a common abiotic stress in tropical and subtropical regions that limits the growth and development of agricultural crops, mainly impacting grain yield. Acting through plant breeding is the most viable alternative for obtaining genotypes more tolerant of environments with stress. This work aims to select popcorn genotypes for environments with drought and to identify discriminating traits for the evaluation of drought tolerance in popcorn germplasm. Fifteen Latin American populations of popcorn were evaluated in water stress (WS) and well-watered (WW) conditions. The evaluated traits were based in morpho-agronomic, physiological and radicular descriptors. Data were submitted to individual and joint ANOVA and GT Biplot analysis. Variability was detected between populations for all traits in both conditions. The drought caused a reduction of 30.61% and 3.5% in grain yield and popping expansion, respectively. Based in GT biplot analysis, 880POP was the most stable in WS and WW, being indicated as a promising population for cultivation in environments with water limitation. This study is going to allow the establishment of a collection of great importance to maize germplasm and to provide information to facilitate the process of selection in breeding programs focused on drought tolerance.

Genome-wide association mapping reveals new loci associated with light-colored seed coat at harvest and slow darkening in carioca beans.

BACKGROUND: Common bean (Phaseolus vulgaris L.) is a legume whose grain can be stored for months, a common practice among Brazilian growers. Over time, seed coats become darker and harder to cook, traits that are undesirable to consumers, who associate darker-colored beans with greater age. Like commercial pinto and cranberry bean varieties, carioca beans that have darker seeds at harvest time and after storage are subject to decreased market values. RESULTS: The goal of our study was to identify the genetic control associated with lightness of seed coat color at harvest (HL) and with tolerance to post-harvest seed coat darkening (PHD) by a genome-wide association study. For that purpose, a carioca diversity panel previously validated for association mapping studies was used with 138 genotypes and 1,516 high-quality SNPs. The panel was evaluated in two environments using a colorimeter and the CIELAB scale. Shelf storage for 30 days had the most expressive results and the L* (luminosity) parameter led to the greatest discrimination of genotypes. Three QTL were identified for HL, two on chromosome Pv04 and one on Pv10. Regarding PHD, results showed that genetic control differs for L* after 30 days and for the ΔL* (final L*-initial L*); only ΔL* was able to properly express the PHD trait. Four phenotypic classes were proposed, and five QTL were identified through six significant SNPs. CONCLUSIONS: Lightness of seed coat color at harvest showed an oligogenic inheritance corroborated by moderate broad-sense heritability and high genotypic correlation among the experiments. Only three QTL were significant for this trait - two were mapped on Pv04 and one on Pv10. Considering the ΔL, six QTL were mapped on four different chromosomes for PHD. The same HL QTL at the beginning of Pv10 was also associated with ΔL* and could be used as a tool in marker-assisted selection. Several candidate genes were identified and may be useful to accelerate the genetic breeding process.

IFNγ Modulates the Immunopeptidome of Triple Negative Breast Cancer Cells by Enhancing and Diversifying Antigen Processing and Presentation.

Peptide vaccination remains a viable approach to induce T-cell mediated killing of tumors. To identify potential T-cell targets for Triple-Negative Breast Cancer (TNBC) vaccination, we examined the effect of the pro-inflammatory cytokine interferon-γ (IFNγ) on the transcriptome, proteome, and immunopeptidome of the TNBC cell line MDA-MB-231. Using high resolution mass spectrometry, we identified a total of 84,131 peptides from 9,647 source proteins presented by human leukocyte antigen (HLA)-I and HLA-II alleles. Treatment with IFNγ resulted in a remarkable remolding of the immunopeptidome, with only a 34% overlap between untreated and treated cells across the HLA-I immunopeptidome, and expression of HLA-II only detected on treated cells. IFNγ increased the overall number, diversity, and abundance of peptides contained within the immunopeptidome, as well increasing the coverage of individual source antigens. The suite of peptides displayed under conditions of IFNγ treatment included many known tumor associated antigens, with the HLA-II repertoire sampling 17 breast cancer associated antigens absent from those sampled by HLA-I molecules. Quantitative analysis of the transcriptome (10,248 transcripts) and proteome (6,783 proteins) of these cells revealed 229 common proteins and transcripts that were differentially expressed. Most of these represented downstream targets of IFNγ signaling including components of the antigen processing machinery such as tapasin and HLA molecules. However, these changes in protein expression did not explain the dramatic modulation of the immunopeptidome following IFNγ treatment. These results demonstrate the high degree of plasticity in the immunopeptidome of TNBC cells following cytokine stimulation and provide evidence that under pro-inflammatory conditions a greater variety of potential HLA-I and HLA-II vaccine targets are unveiled to the immune system. This has important implications for the development of personalized cancer vaccination strategies.

Genome-wide association mapping reveals race-specific SNP markers associated with anthracnose resistance in carioca common beans.

Brazil is the largest consumer of dry edible beans (Phaseolus vulgaris L.) in the world, 70% of consumption is of the carioca variety. Although the variety has high yield, it is susceptible to several diseases, among them, anthracnose (ANT) can lead to losses of up to 100% of production. The most effective strategy to overcome ANT, a disease caused by the fungus Colletotrichum lindemuthianum, is the development of resistant cultivars. For that reason, the selection of carioca genotypes resistant to multiple ANT races and the identification of loci/markers associated with genetic resistance are extremely important for the genetic breeding process. Using a carioca diversity panel (CDP) with 125 genotypes and genotyped by BeadChip BARCBean6K_3 and a carioca segregating population AM (AND-277 × IAC-Milênio) genotyped by sequencing (GBS). Multiple interval mapping (MIM) and genome-wide association studies (GWAS) were used as mapping tools for the resistance genes to the major ANT physiological races present in the country. In general, 14 single nucleotide polymorphisms (SNPs) showed high significance for resistance by GWAS, and loci associated with multiple races were also identified, as the Co-3 locus. The SNPs ss715642306 and ss715649427 in linkage disequilibrium (LD) at the beginning of chromosome Pv04 were associated with all the races used, and 16 genes known to be related to plant immunity were identified in this region. Using the resistant cultivars and the markers associated with significant quantitative resistance loci (QRL), discriminant analysis of principal components (DAPC) was performed considering the allelic contribution to resistance. Through the DAPC clustering, cultivar sources with high potential for durable anthracnose resistance were recommended. The MIM confirmed the presence of the Co-14 locus in the AND-277 cultivar which revealed that it was the only one associated with resistance to ANT race 81. Three other loci were associated with race 81 on chromosomes Pv03, Pv10, and Pv11. This is the first study to identify new resistance loci in the AND-277 cultivar. Finally, the same Co-14 locus was also significant for the CDP at the end of Pv01. The new SNPs identified, especially those associated with more than one race, present great potential for use in marker-assisted and early selection of inbred lines.