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Abstract 3D skin models have been explored as an alternative method to the use of animals in research and development. Usually, human skin equivalents comprise only epidermis or epidermis/dermis layers. Herein, we leverage 3D bioprinting technology to fabricate a full-thickness human skin equivalent with hypodermis (HSEH). The collagen hydrogel-based structure provides a mimetic environment for skin cells to adhere, proliferate and differentiate. The effective incorporation of the hypodermis layer is evidenced by scanning electron microscopy, immunofluorescence, and hematoxylin and eosin staining. The transcriptome results underscore the pivotal role of the hypodermis in orchestrating the genetic expression of a multitude of genes vital for skin functionality, including hydration, development and differentiation. Accordingly, we evidence the paramount significance of full-thickness human skin equivalents with hypodermis layer to provide an accurate in vitro platform for disease modeling and toxicology studies.

Abstract Glutaminase (GLS) is directly related to cell growth and tumor progression, making it a target for cancer treatment. The RNA-binding protein HuR (encoded by the ELAVL1 gene) influences mRNA stability and alternative splicing. Overexpression of ELAVL1 is common in several cancers, including breast cancer. Here we show that HuR regulates GLS mRNA alternative splicing and isoform translation/stability in breast cancer. Elevated ELAVL1 expression correlates with high levels of the glutaminase isoforms C (GAC) and kidney-type (KGA), which are associated with poor patient prognosis. Knocking down ELAVL1 reduces KGA and increases GAC levels, enhances glutamine anaplerosis into the TCA cycle, and drives cells towards glutamine dependence. Furthermore, we show that combining chemical inhibition of GLS with ELAVL1 silencing synergistically decreases breast cancer cell growth and invasion. These findings suggest that dual inhibition of GLS and HuR offers a therapeutic strategy for breast cancer treatment.

Background Despite advances in cancer care and detection, >65% of patients with squamous cell cancer of the head and neck (HNSCC) will develop recurrent and/or metastatic disease. The prognosis for these patients is poor with a 5-year overall survival of 39%. Recent treatment advances in immunotherapy, including immune checkpoint inhibitors like pembrolizumab and nivolumab, have resulted in clinical benefit in a subset of patients. There is a critical clinical need to identify patients who benefit from these antiprogrammed cell death protein 1 (anti-PD-1) immune checkpoint inhibitors.Methods Here, we report findings from a multicenter observational study, PREDicting immunotherapy efficacy from Analysis of Pre-treatment Tumor biopsies (PREDAPT), conducted across 17 US healthcare systems. PREDAPT aimed to validate OncoPrism-HNSCC, a clinical biomarker assay predictive of disease control in patients with recurrent or metastatic HNSCC treated with anti-PD-1 immune checkpoint inhibitors as a single agent (monotherapy) and in combination with chemotherapy (chemo-immunotherapy). The test used RNA-sequencing data and machine learning models to score each patient and place them into groups of low, medium, or high.Results The OncoPrism-HNSCC prediction significantly correlated with disease control in both the monotherapy cohort (n=62, p=0.004) and the chemo-immunotherapy cohort (n=50, p=0.01). OncoPrism-HNSCC also significantly predicted progression-free survival in both cohorts (p=0.015 and p=0.037, respectively). OncoPrism-HNSCC had more than threefold higher specificity than programmed death-ligand 1 combined positive score and nearly fourfold higher sensitivity than tumor mutational burden for predicting disease control.Conclusions Here, we demonstrate the clinical validity of the OncoPrism-HNSCC assay in identifying patients with disease control in response to anti-PD-1 immune checkpoint inhibitors.Trial registration number NCT04510129.

Abstract Cystathionine beta-synthase (CBS) is an essential metabolic enzyme across all domains of life for the production of glutathione, cysteine, and hydrogen sulfide. Appended to the conserved catalytic domain of human CBS is a regulatory domain that modulates activity by S-adenosyl-L-methionine (SAM) and promotes oligomerisation. Here we show using cryo-electron microscopy that full-length human CBS in the basal and SAM-bound activated states polymerises as filaments mediated by a conserved regulatory domain loop. In the basal state, CBS regulatory domains sterically block the catalytic domain active site, resulting in a low-activity filament with three CBS dimers per turn. This steric block is removed when in the activated state, one SAM molecule binds to the regulatory domain, forming a high-activity filament with two CBS dimers per turn. These large conformational changes result in a central filament of SAM-stabilised regulatory domains at the core, decorated with highly flexible catalytic domains. Polymerisation stabilises CBS and reduces thermal denaturation. In PC-3 cells, we observed nutrient-responsive CBS filamentation that disassembles when methionine is depleted and reversed in the presence of SAM. Together our findings extend our understanding of CBS enzyme regulation, and open new avenues for investigating the pathogenic mechanism and therapeutic opportunities for CBS-associated disorders.

Accuracy and transparency of scientific data are becoming more and more relevant with the increasing concern regarding the evaluation of data reproducibility in many research areas. This concern is also true for quantifying coding and noncoding RNAs, with the remarkable increase in publications reporting RNA profiling and sequencing studies. To address the problem, we propose the following recommendations: (a) accurate documentation of experimental procedures in Materials and methods (and not only in the supplementary information, as many journals have a strict mandate for making Materials and methods as visible as possible in the main text); (b) submission of RT‐qPCR raw data for all experiments reported; and (c) adoption of a unified, simple format for submitted RT‐qPCR raw data. The Real‐time PCR Data Essential Spreadsheet Format (RDES) was created for this purpose.

Analysing the regulatory consequences of mutations and splice variants at large scale in cancer requires efficient computational tools. Here, the authors develop RegTools, a software package that can identify splice-associated variants from large-scale genomics and transcriptomics data with efficiency and flexibility.

One of the challenging problems in large scale cyber-argumentation platforms is that users often engage and focus only on a few issues and leave other issues under-discussed and under-acknowledged. This kind of non-uniform participation obstructs the argumentation analysis models to retrieve collective intelligence from the underlying discussion. To resolve this problem, we developed an innovative opinion prediction model for a multi-issue cyber-argumentation environment. Our model predicts users’ opinions on the non-participated issues from similar users’ opinions on related issues using intelligent argumentation techniques and a collaborative filtering method. Based on our detailed experimental results on an empirical dataset collected using our cyber-argumentation platform, our model is 21.7% more accurate, handles data sparsity better than other popular opinion prediction methods. Our model can also predict opinions on multiple issues simultaneously with reasonable accuracy. Contrary to existing opinion prediction models, which only predict whether a user agrees on an issue, our model predicts how much a user agrees on the issue. To our knowledge, this is the first research to attempt multi-issue opinion prediction with the partial agreement in the cyber-argumentation platform. With additional data on non-participated issues, our opinion prediction model can help the collective intelligence analysis models to analyze social phenomena more effectively and accurately in the cyber argumentation platform.

ABSTRACT Global warming potential (GWP) of rice paddies depends on straw management. This study evaluated methane (CH4) and nitrous oxide (N2O) emissions and soil C stocks to determine GWP and yield-scaled GWP under different strategies and intensities of rice straw management in a subtropical climate. We hypothesized that decreasing soil management intensity and straw incorporation in the soil would reduce GWP. Methane fluxes were substantially higher during the rice growing season than in the off-season, while the opposite was observed for N2O fluxes. The cumulative emissions of CH4 during the growing season among the straw management strategies evaluated ranged from 165.8 to 586.0 kg ha-1. Annual CH4 emissions were lower when soil and straw received some type of management compared to no-tillage. Daily N2O fluxes ranged from -2.8 to 201.7 g ha-1 day-1; cumulative N2O emissions during the off-season ranged from 455.2 to 2816.5 g ha-1. During the off-season, strategies to reduce N2O emissions include post-harvest straw incorporation using a disc harrow, winter straw removal, and ryegrass cropping. Soil organic C stocks ranged from 35.96 to 38.36 Mg ha-1. Straw management using a disc harrow reduced soil organic C stocks, with more adverse effects than straw removal. Soil and rice straw management did not affect rice grain yield, with an average of 10.4 Mg ha-1. Methane emissions were the main component of GWP in all straw management systems. The contribution of N2O emissions to GWP was small and mostly (>85 %) determined by off-season emissions. Yield-scaled GWP ranged from 0.64 to 1.06 Mg CO2eq Mg-1 yield and was lower when soil and straw management systems occurred shortly after the rice harvest. Our results indicate that soil and straw management immediately after rice harvest reduces CH4 emissions, GWP, and yield-scaled GWP.

Background and study aims Hemospray (TC-325) is a mineral powder with adsorptive properties designed for use in various gastrointestinal bleeding (GIB) scenarios. We conducted a systematic review & meta-analysis of randomized controlled trials (RCTs) comparing TC-325 to standard endoscopic therapy (SET) for non-variceal GIB (NVGIB). Methods Multiple databases were searched through October 2022. Meta-analysis was performed using a random-effects model to determine pooled relative risk (RR) and proportions with 95 % confidence intervals (CI) for primary hemostasis, hemostasis failure, 30-day rebleeding, length of stay (LOS), and need for rescue interventions. Heterogeneity was assessed using I2%. Results Five RCTs with 362 patients (TC-325 178, SET 184) – 123 females and 239 males with a mean age 65 ± 16 years). The most common etiologies were peptic ulcer disease (48 %), malignancies (35 %), and others (17 %). Bleeding was characterized as Forrest IA (7 %), IB (73 %), IIA (3 %), and IIB (1 %). SET included epinephrine injection, electrocautery, hemoclips, or a combination. No statistical difference in primary hemostasis between TC-325 compared to SET, RR 1.09 (CI 0.95–1.25; I2 43), P = 0.2, including patients with oozing/spurting hemorrhage, RR 1.13 (CI 0.98–1.3; I2 35), P = 0.08. Failure to achieve hemostasis was higher in SET compared to TC-325, RR 0.30 (CI 0.12–0.77, I2 0), P = 0.01, including patients with oozing/spurting hemorrhage, RR 0.24 (CI 0.09 – 0.63, I2 0), P = 0.004. We found no difference between the two interventions in terms of rebleeding, RR 1.13 (CI 0.62–2.07, I2 26), P = 0.8 and LOS, standardized mean difference (SMD) 0.27 (CI, –0.20–0.74; I2 62), P = 0.3. Finally, pooled rate of rescue interventions (angiography) was statistically higher in SET compared to TC-325, RR 0.68 (CI 0.5–0.94; I2 0), P = 0.02. Conclusions Our analysis shows that for acute NV GIB, including oozing/spurting hemorrhage, TC-325 does not result in higher rates of primary hemostasis compared to SET. However, lower rates of failures were seen with TC-325 than SET. In addition, there was no difference in the two modalities when comparing rates of rebleeding and LOS.

After the Coronavirus pandemic, the importance of virus surveillance was highlighted, reinforcing the constant necessity of discussing and updating the methods for collection and diagnoses, including for other respiratory viruses. Although the nasopharyngeal swab is the gold-standard sample for detecting and genotyping SARS-CoV-2 and Influenza viruses, its collection is uncomfortable and requires specialized teams, which can be costly. During the pandemic, non-invasive saliva samples proved to be a suitable alternative for SARS-CoV-2 diagnosis, but for Influenza virus the use of this sample source is not recognized yet. In addition, most SARS-CoV-2 comparisons were conducted before the Omicron variant emerged. Here, we aimed to compare Influenza A and Omicron RT-qPCR analysis of nasopharyngeal swabs and saliva self-collection in paired samples from 663 individuals. We found that both nasopharyngeal swab and saliva collection are efficient for the diagnosis of Omicron (including sub-lineages) and for Influenza A, with high sensitivity and accuracy (>90%). The kappa index is 0.938 for Influenza A and 0.905 for SARS-CoV-2. These results showed excellent agreement between the two samples reinforcing saliva samples as a reliable source for detecting Omicron and highlighting saliva as a valid sample source for Influenza detection, considering this cheaper and more comfortable alternative.

Abstract Anti-PD-1 therapy can provide long, durable benefit to a fraction of patients. The on-label PD-L1 test, however, does not accurately predict response. To build a better biomarker, we created a method called T Cell Subtype Profiling (TCSP) that characterizes the abundance of T cell subtypes (TCSs) in FFPE specimens using five RNA models. These TCS RNA models are created using functional methods, and robustly discriminate between naïve, activated, exhausted, effector memory, and central memory TCSs, without the reliance on non-specific, classical markers. TCSP is analytically valid and corroborates associations between TCSs and clinical outcomes. Multianalyte biomarkers based on TCS estimates predicted response to anti-PD-1 therapy in three different cancers and outperformed the indicated PD-L1 test, as well as Tumor Mutational Burden. Given the utility of TCSP, we investigated the abundance of TCSs in TCGA cancers and created a portal to enable researchers to discover other TCSP-based biomarkers.

Cancer cells rely on different mechanisms to maintain their dysregulated metabolism and high proliferation. Metabolic rewiring with an increased dependency on glutamine, the most abundant amino acid in the blood, is a common feature of several types of cancers, including triple-negative breast cancer (TNBC). The enzyme glutaminase (GLS) converts glutamine to glutamate, a reaction that provides α-ketoglutarate for the tricarboxylic/citric acid cycle (TCA) and chromatin-modifying enzymes, thus impacting proliferation and cell differentiation. Glutamine metabolism also impacts the redox balance and synthesis of nucleotides, lipids, and other amino acids. Glutaminase is a potential therapeutic target for cancer, with inhibitors in clinical trials. We previously performed high-throughput screening (HTS) to look for GLS inhibitors. Here, we present the structure–activity relationship (SAR) study of one of the hits, C12, a molecule containing the heteroaromatic nucleus 2-amino-thiazole. The SAR investigation was carried out through modifications of the substituents from the main 3 rings present at the C12 prototype, rings A-C. The combination of 4-F and phenylacetic substitutions on ring A, the incorporation of a 1,3,4-thiadiazole moiety on ring B, and a 4-CN substituent on ring C were the best options to improve specific GLS inhibition. In this series, three compounds exhibited suitable enzyme (GAC vs GLS2) and cell (MDA-MB-231 vs SKBR3) selectivity and, thus, could be useful for the development of new glutaminase inhibitors.

We performed a large-scale severe acute respiratory syndrome coronavirus 2 screening campaign using 2 PCR-based approaches, coupled with variant genotyping, aiming to provide a safer environment for employees of Federal University in Curitiba, Brazil. We observed the rapid spread of the Gamma variant of concern, which replaced other variants in

A toxoplasmose é uma doença cosmopolita, causada pelo Toxoplasma gondii, com grande importância no contexto de produção animal e saúde pública como evidenciado nos dois maiores surtos da doença que ocorreram nas cidades brasileiras de em Santa Izabel do Ivai-PR e Santa Maria-RS. Enquanto o Neospora caninum é o agente causador da neosporose e a Brucella spp. causadora da brucelose, duas das principais causas de falhas reprodutivas em ruminantes, ocasionando problemas como abortamento, e responsáveis por significativas perdas econômicas na ovinocultura brasileira. Considerando a escassez de dados disponíveis sobre o tema para a região estudada, o objetivo do presente estudo foi avaliar a soroprevalência de anticorpos contra T. gondii, N. caninum e B. abortus em ovinos do município de Toledo, Paraná, Brasil. Durante o período de agosto de 2017 a julho de 2018 foram visitadas 22 propriedades e coletadas 240 amostras de sangue de fêmeas e machos em idade reprodutiva. Para a detecção de anticorpos contra o T. gondii e N. caninum foi utilizada a técnica de Imunofluorescência Indireta empregando ponto de corte ≥64 e ≥50, respectivamente. Para a detecção de anticorpos contra B. abortus foi realizada a triagem com o Antígeno Acidificado Tamponado e confirmação das amostras reagentes com o teste de Soroaglutinação Lenta e do 2-Mercaptoetanol. Das amostras analisadas, 27,08% (65/240) apresentaram anticorpos contra o T. gondii, estes se encontravam distribuídas em 81,82% (18/22) das propriedades, para N. caninum observou-se uma prevalência de 15,41% (37/240), distribuídas em 68,18% (15/22) das propriedades. Quanto ao resultado dos testes da B. abortus todas as amostras foram consideradas negativas. O presente estudo mostrou que a grande maioria das propriedades apresentaram animais positivos para os protozoários, os quais, principalmente T. gondii, poderiam estar causando problemas de abortamentos. Estes dados epidemiológicos fornecem embasamento para programas estratégicos a fim de realizar medidas de monitoramento e controle, evitando perdas econômicas na ovinocultura.

Background and study aims Endoscopic retrograde appendicitis therapy (ERAT) is an endoscopic procedure for management of patients with acute appendicitis (AA). In addition to being minimally invasive, it has the added advantages of preservation of appendix and simultaneous inspection of colon. We performed a systematic review and meta-analysis on ERAT in patients with AA. Methods We conducted a comprehensive search of multiple electronic databases (from inception through January 2022) to identify studies reporting ERAT in AA. The primary outcome was to evaluate the overall clinical and technical success of ERAT. The secondary outcome was to study the total and individual adverse events (AEs). The meta-analysis was performed using Der Simonian and Laird random effect model. Results Seven studies reporting on 298 patients were included. The majority of the patient population was male (55.3 %), with mean age of 31 ± 12.39 years. The pooled technical success rate was 99.36 % (95 % CI 97.61–100, I2 = 0) and the pooled clinical success rate was 99.29 % (95 % CI 97.48–100, I2 = 0). The pooled AE rate was 0.19 % (95 % CI 0–1.55, I2 = 0). The most common AE was perforation with 0.19 % (95 % CI 0–1.55, I2 = 0). The recurrence rate was 6.01 % (95 % CI 2.9–9.93, I2 = 20.10). Average length of procedure was 41.1 ± 7.16 min. Low heterogeneity was noted in in our meta-analysis. Conclusions ERAT is a safe procedure with high rates of clinical and technical success in patients with AA. Further randomized controlled trials should be performed to assess the utility of ERAT in AA as compared to laparoscopic appendectomy.

Breast cancer (BC) is a heterogeneous disease, and establishing biomarkers is essential to patient management. We previously described that extracellular vesicle–derived miRNAs (EV-miRNAs) miR-142-5p, miR-150-5p, miR-320a, and miR-4433b-5p in serum discriminated BC from control samples, either alone or combined in a panel. Using these previously described markers, we intend to evaluate whether the same markers identified in EVs are also potential biomarkers in tissue and serum. Expression analysis using RT-qPCR was performed using serum of 67 breast cancer patients (BC-S), 19 serum controls (CT), 83 fresh tumor tissues (BC-T), and 29 adjacent nontumor tissue samples (NT). In addition, analysis from The Cancer Genome Atlas (TCGA) data (832 BC-T and 136 NT) was performed. In all comparisons, we found concordant high expression levels of miR-320a and miR-4433b-5p in BC-S compared to CT in both EVs and cell-free miRNAs (cf-miRNAs). Although miR-150-5p and miR-142-5p were not found to be differentially expressed in serum, panels including these miRNAs improved sensitivity and specificity, supporting our previous findings in EVs. Fresh tissue and data from the TCGA database had, in most comparisons, an opposite behavior when compared to serum and EVs: lower levels of all miRNAs in BC-T than those in NT samples. TCGA analyses revealed reduced expression levels of miR-150-5p and miR-320a-3p in BC-T than those in NT samples and the overexpression of miR-142-5p in BC-T, unlike our RT-qPCR results from tissue in the Brazilian cohort. The fresh tissue analysis showed that all miRNAs individually could discriminate between BC-T and NT in the Brazilian cohort, with high sensitivity and sensibility. Furthermore, combining panels showed higher AUC values and improved sensitivity and specificity. In addition, lower levels of miR-320a-3p in serum were associated with poor overall survival in BC Brazilian patients. In summary, we observed that miR-320a and miR-4433b-5p distinguished BC from controls with high specificity and sensibility, regardless of the sample source. In addition, lower levels of miR-150-5p and higher levels of miR-142-5p were statistically significant biomarkers in tissue, according to TCGA. When combined in panels, all combinations could distinguish BC patients from controls. These results highlight a potential application of these miRNAs as BC biomarkers.

Abstract Background Xentuzumab—a humanised IgG1 monoclonal antibody—binds IGF-1 and IGF-2, inhibiting their growth-promoting signalling and suppressing AKT activation by everolimus. This phase Ib/II exploratory trial evaluated xentuzumab plus everolimus and exemestane in hormone receptor-positive, locally advanced and/or metastatic breast cancer (LA/MBC). Methods Patients with hormone receptor-positive/HER2-negative LA/MBC resistant to non-steroidal aromatase inhibitors were enrolled. Maximum tolerated dose (MTD) and recommended phase II dose (RP2D) of xentuzumab/everolimus/exemestane were determined in phase I (single-arm, dose-escalation). In phase II (open-label), patients were randomised 1:1 to the RP2D of xentuzumab/everolimus/exemestane or everolimus/exemestane alone. Randomisation was stratified by the presence of visceral metastases. Primary endpoint was progression-free survival (PFS). Results MTD was determined as xentuzumab 1000 mg weekly plus everolimus 10 mg/day and exemestane 25 mg/day. A total of 140 patients were enrolled in phase II (70 to each arm). Further recruitment was stopped following an unfavourable benefit-risk assessment by the internal Data Monitoring Committee appointed by the sponsor. Xentuzumab was discontinued; patients could receive everolimus/exemestane if clinically indicated. Median PFS was 7.3 months (95% CI 3.3–not calculable) in the xentuzumab/everolimus/exemestane group and 5.6 months (3.7–9.1) in the everolimus/exemestane group (hazard ratio 0.97, 95% CI 0.57–1.65; P = 0.9057). In a pre-specified subgroup of patients without visceral metastases at screening, xentuzumab/everolimus/exemestane showed evidence of PFS benefit versus everolimus/exemestane (hazard ratio 0.21 [0.05–0.98]; P = 0.0293). Most common any-cause adverse events in phase II were diarrhoea (29 [41.4%] in the xentuzumab/everolimus/exemestane group versus 20 [29.0%] in the everolimus/exemestane group), mucosal inflammation (27 [38.6%] versus 21 [30.4%]), stomatitis (24 [34.3%] versus 24 [34.8%]), and asthenia (21 [30.0%] versus 24 [34.8%]). Conclusions Addition of xentuzumab to everolimus/exemestane did not improve PFS in the overall population, leading to early discontinuation of the trial. Evidence of PFS benefit was observed in patients without visceral metastases when treated with xentuzumab/everolimus/exemestane, leading to initiation of the phase II XENERA™-1 trial (NCT03659136). Trial registration ClinicalTrials.gov, NCT02123823 . Prospectively registered, 8 March 2013.

Summary: Although tobacco use is an independent adverse prognostic feature in HPV(+) oropharyngeal squamous cell carcinoma (OPSCC), the biologic features associated with tobacco use have not been systematically investigated. We characterized genomic and immunologic features associated with tobacco use through whole exome sequencing, mRNA hybridization, and immunohistochemical staining in 47 HPV(+) OPSCC tumors. Low expression of transcripts in a T cell-inflamed gene expression profile (TGEP) was associated with tobacco use at diagnosis and lower overall and disease-free survival. Tobacco use was associated with an increased proportion of T > C substitutions and a lower proportion of expected mutational signatures, but not with increases in mutational burden or recurrent oncogenic mutations. Our findings suggest that rather than increased mutational burden, tobacco’s primary and clinically relevant association in HPV(+) OPSCC is immunosuppression of the tumor immune microenvironment. Quantitative assays of T cell infiltration merit further study as prognostic markers in HPV(+) OPSCC.

The study aims to analyze sustainable entrepreneurship and resonant and innovative leadership as a strategic binomial for sustainable development in Latin American territories, entrepreneurial actions for sustainability must be directed towards innovation, environmental conservation, for the creation of goods and services, improve the quality of life of citizens, it is essential to incorporate resonant and innovative leadership, their optimistic, balanced and comprehensive ability to envision the organization allows timely evaluation of social, political, economic, legal, intercultural or technological aspects, it is important That the leader focuses his attention on the workers since they are the most valuable asset they have, and have capacities and skills that contribute to the achievement of proposed goals, fosters a pleasant work environment, and optimal communication and interaction between workers, this entails gener ar competitive advantages and be highly productive, leading to sustainable development in Latin American territories. Methodologically, it is guided by the quantitative paradigm, with a descriptive scope; non-experimental transactional field, population 25 subjects among managers, supervisors of public service undertakings of the Baralt municipality, Venezuela. 1 instrument containing 39 items was designed following the Likert model with a scale of 5 response alternatives, to analyze the innovative resonant leadership variable, the systematic literature review technique was also used that allowed describing the context of sustainable development in the Latin American territory, through documentary analysis and interpretation, in order to achieve the objective proposed in this study. The results of the study indicate that the main quality of the resonant and innovative leadership referred to that they connect with the feelings of the people, directing them emotionally in a positive way is present in a moderate way, likewise it was evidenced that the leaders are not paying relevant attention to their personal and downplay their capacities and abilities, therefore these are used moderately, which represents serious difficulties for sustainable enterprises, showing that it is highly relevant to consider entrepreneurship and leadership as a strategic binomial for sustainable development.

ABSTRACT BACKGROUND: We aimed to describe the clinical characteristics of coronavirus disease 2019 (COVID-19) among healthcare workers (HCWs) in Curitiba, Brazil. METHODS: Upper respiratory samples from 1077 HCWs were tested for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using reverse transcription polymerase chain reaction from June 16, 2020 to December 9, 2020. RESULTS: Overall, 32.7% of HCWs were infected. The positivity rates in symptomatic and asymptomatic HCWs were 39.2% and 15.9%, respectively. Hospital departments categorized as high-risk for exposure had the highest number of infected HCWs. CONCLUSIONS: Early diagnosis and isolation of infected HCWs remain key in controlling SARS-CoV-2 transmission because HCWs in close contact with COVID-19 patients are more likely to be infected than those who are not.

Abstract Background In patients who have had axillary nodal metastasis diagnosed prior to neoadjuvant chemotherapy for breast cancer, there is little consensus on how to manage the axilla subsequently. The aim of this study was to explore whether a combination of breast magnetic resonance imaging (MRI) assessed response and primary tumour pathology factors could identify a subset of patients that might be spared axillary node clearance. Methods A retrospective data analysis was performed of patients with core biopsy-proven axillary nodal metastasis prior to commencement of neoadjuvant chemotherapy (NAC) who had subsequent axillary node clearance (ANC) at definitive breast surgery. Breast tumour and axillary response at MRI before, during and on completion of NAC, core biopsy tumour grade, tumour type and immunophenotype were correlated with pathological response in the breast and the number of metastatic nodes in the ANC specimens. Results Of 87 consecutive patients with MRI at baseline, interim and after neoadjuvant chemotherapy who underwent ANC at time of breast surgery, 33 (38%) had no residual macrometastatic axillary disease, 28 (32%) had 1–2 metastatic nodes and 26 (30%) had more than 2 metastatic nodes. Factors that predicted axillary nodal complete response were MRI complete response in the breast (p

A methodology is proposed to incorporate spatial distributions of micro-defects within a three dimensional multiscale modeling framework for Carbon Fiber Reinforced Polymer (CFRP) composites to predict stiffness and strength properties of laminates for use in structural design and in establishing nondestructive inspection protocols. Defects in both the fiber and matrix due to variations in the manufacturing process for fiber reinforced composite laminates are considered. Stochastic sampling of micro-defects based on experimental data using Ripley’s K-function is proposed. Three different laminate void volume fractions are used. Spatial statistical analysis is performed for the micro-defects, and then a methodology is presented for predicting stiffness and ultimate strength of three multi-directional composite laminates with distributions of microstructural defects. The results indicate that, in addition to the volume of micro-defects that are present, the spatial distribution of the micro-defects plays a significant role in mechanical properties of laminates, especially the ultimate strength for which a 3.62% reduction is seen in the simulations of laminates containing distributions of micro-defects. Based on these results, it is recommended that the spatial statistics of micro-defects in manufactured composites be incorporated into both design criteria and nondestructive inspection criteria for composite laminates.

ABSTRACT: Crop residues decomposition are controlled by chemical tissue components. This study evaluated changes on plant tissue components, separated by the Van Soest partitioning method, during cover crop decomposition. The Van Soest soluble fraction was the first to be released from the crop residues, followed by cellulose and hemicellulose. Lignin was the crop residue component that suffered the least degradation, and for certain crop residue types, lignin degradation was not detected. The degradation of the main components of crop residues (soluble fraction, cellulose, hemicellulose and lignin) is determined by the chemical and structural composition of each fraction.

Background: Most patients with oesophageal cancer present with incurable disease. For those with advanced disease, the mean survival is 3–5 months. Treatment emphasis is therefore on effective palliation, with the majority of patients requiring intervention for dysphagia. Insertion of a self-expanding metal stent provides rapid relief but dysphagia may recur within 3 months owing to tumour progression. Evidence reviews have called for trials of interventions combined with stenting to better maintain the ability to swallow. Objectives: The Radiotherapy after Oesophageal Cancer Stenting (ROCS) study examined the effectiveness of palliative radiotherapy, combined with insertion of a stent, in maintaining the ability to swallow. The trial also examined the impact that the ability to swallow had on quality of life, bleeding events, survival and cost-effectiveness. Design: A pragmatic, multicentre, randomised controlled trial with follow-up every 4 weeks for 12 months. An embedded qualitative study examined trial experiences in a participant subgroup. Setting: Participants were recruited in secondary care, with all planned follow-up at home. Participants: Patients who were referred for stent insertion as the primary management of dysphagia related to incurable oesophageal cancer. Interventions: Following stent insertion, the external beam radiotherapy arm received palliative oesophageal radiotherapy at a dose of 20 Gy in five fractions or 30 Gy in 10 fractions. Main outcome measures: The primary outcome was the difference in the proportion of participants with recurrent dysphagia, or death, at 12 weeks. Recurrent dysphagia was defined as deterioration of ≥ 11 points on the dysphagia scale of the European Organisation of Research and Treatment of Cancer Quality of Life Questionnaire oesophago-gastric module questionnaire. Secondary outcomes included quality of life, bleeding risk and survival. Results: The study recruited 220 patients: 112 were randomised to the usual-care arm and 108 were randomised to the external beam radiotherapy arm. There was no evidence that radiotherapy reduced recurrence of dysphagia at 12 weeks (48.6% in the usual-care arm compared with 45.3% in the external beam radiotherapy arm; adjusted odds ratio 0.82, 95% confidence interval 0.40 to 1.68; p = 0.587) and it was less cost-effective than stent insertion alone. There was no difference in median survival or key quality-of-life outcomes. There were fewer bleeding events in the external beam radiotherapy arm. Exploration of patient experience prompted changes to trial processes. Participants in both trial arms experienced difficulty in managing the physical and psychosocial aspects of eating restriction and uncertainties of living with advanced oesophageal cancer. Limitations: Change in timing of the primary outcome to 12 weeks may affect the ability to detect a true intervention effect. However, consistency of results across sensitivity analyses is robust, including secondary analysis of dysphagia deterioration-free survival. Conclusions: Widely accessible palliative external beam radiotherapy in combination with stent insertion does not reduce the risk of dysphagia recurrence at 12 weeks, does not have an impact on survival and is less cost-effective than inserting a stent alone. Reductions in bleeding events should be considered in the context of patient-described trade-offs of fatigue and burdens of attending hospital. Trial design elements including at-home data capture, regular multicentre nurse meetings and qualitative enquiry improved recruitment/data capture, and should be considered for future studies. Future work: Further studies are required to identify interventions that improve stent efficacy and to address the multidimensional challenges of eating and nutrition in this patient population. Trial registration: Current Controlled Trials ISRCTN12376468 and Clinicaltrials.gov NCT01915693. Funding: This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 25, No. 31. See the NIHR Journals Library website for further project information.

Screening efforts and genomic surveillance are essential tools to evaluate the course of the COVID-19 pandemic and assist the public healthcare system in dealing with an increasing number of infections. For the analysis of COVID-19 cases scenarios in Curitiba, Paraná, Brazil, we performed a diagnosis of positive cases, coupled with genotyping, for symptomatic and asymptomatic members of the Federal University of Paraná. We achieved over 1000 samples using RT-qPCR for diagnosis. The posterior genotyping allowed us to observe differences in the spread of strains in Curitiba, Brazil. The Delta variant was not associated with an infection wave, whereas the rapid Omicron variant spread became dominant in less than one month. We also evaluated the general vaccination coverage in the state, observing a striking reduction in lethality correlated to the vaccinated fraction of the population; although lower lethality rates were not much affected by the Omicron variant wave, the same effect was not translated in the number of infections. In summary, our results provide a general overview of the pandemic’s course in Paraná State and how there was reduction in lethality after a combination of multiple infection waves and a large-scale vaccination program.

Background and study aims We compared the diagnostic yield and specimen adequacy in EUS-guided parenchymal biopsies between two types of EUS 19 G core needles. Patients and methods This is a retrospective study of 420 patients at two tertiary medical centers in Florida with unexplained abnormal liver associated tests were referred for EUS evaluation of biliary obstruction and pancreatic pathology. EUS-guided liver biopsy (EUS-LB) was performed at the same session after biliary obstruction was excluded. We compared intact specimen length (ISL), total specimen length (TSL), complete portal triads (CPT) and adverse events (AE). Welch’s T and Tukey tests were used for ISL, TSL and CPT. Results A total of 210 patients underwent EUS-LB using a Franseen needle, 210 patients using a fork-tip needle. Median patient age was 52 years (15.63) and 238 (56.7 %) were female. The fork-tip needle had a mean ISL of 2.7 (1.1 SD) cm, TSL of 6 cm (2.1 SD), and mean 19.5 CPT (8.5 SD) Abdominal pain occurred in 35 patients (17 %) post-procedure and was managed with supportive care. Two patients required intravenous (IV) narcotic administration. Subcapsular hematomas occurred in 1 (0.5 %) patients. The Franseen needle had a mean ISL of 3.1 cm (1.3 SD), TSL of 6.5 cm (2.6 SD), and mean of 24 CPT (8.8 SD). Abdominal pain occurred in four patients (2 %) post-procedure, which resolved in all patients after IV narcotic administration. Subcapsular hematomas occurred in 1 (0.5 %) and bile leak in 1(0.4 %) patients. Conclusions Use of the Franseen needle resulted in better liver core samples than that obtained with a fork-tip needle.

Ultraconserved regions (UCRs) are 481 genome segments, with length longer than 200 bp, that are 100% conserved among humans, mice, and rats. The majority of UCRs are transcriptionally active (T-UCRs) as many of them produce non-coding RNAs. In a previous study, we evaluated the expression level of T-UCRs in breast cancer (BC) patients and found that 63% of transcripts correlated with some clinical and/or molecular parameter of BC. In this study, we delved into the expression levels of 12 T-UCRs and correlated them with clinicopathological parameters, immunohistochemical markers, and overall survival in two breast cancer cohorts: TCGA and Brazilian patients. We found that uc.268 is more expressed in TCGA patients under 40 years of age, associated with progesterone receptor (PR) and estrogen receptor (ER), and its high expression is found in luminal A. Lower uc.84 and uc.376 were respectively observed in metastatic and stage IV tumors associated with good prognostic in luminal B. Moreover, uc.84 was only related to the HER2+, while uc.376 was related to ER+ and PR+, and HER2+. A panel composed of uc.147, uc.271, and uc.427 distinguished luminal A from triple negative patients with an AUC of 0.9531 (sensitivity 92.19% and specificity 86.76%). These results highlight the potential role of T-UCRs in BC and provide insights into the potential application of T-UCRs as biomarkers.

Abstract The active transport of glycolytic pyruvate across the inner mitochondrial membrane is thought to involve two mitochondrial pyruvate carrier subunits, MPC1 and MPC2, assembled as a 150 kDa heterotypic oligomer. Here, the recombinant production of human MPC through a co-expression strategy is first described; however, substantial complex formation was not observed, and predominantly individual subunits were purified. In contrast to MPC1, which co-purifies with a host chaperone, we demonstrated that MPC2 homo-oligomers promote efficient pyruvate transport into proteoliposomes. The derived functional requirements and kinetic features of MPC2 resemble those previously demonstrated for MPC in the literature. Distinctly, chemical inhibition of transport is observed only for a thiazolidinedione derivative. The autonomous transport role for MPC2 is validated in cells when the ectopic expression of human MPC2 in yeast lacking endogenous MPC stimulated growth and increased oxygen consumption. Multiple oligomeric species of MPC2 across mitochondrial isolates, purified protein and artificial lipid bilayers suggest functional high-order complexes. Significant changes in the secondary structure content of MPC2, as probed by synchrotron radiation circular dichroism, further supports the interaction between the protein and ligands. Our results provide the initial framework for the independent role of MPC2 in homeostasis and diseases related to dysregulated pyruvate metabolism.

Abstract Background Triple-negative breast cancer (TNBC) is characterized by a lack of estrogen and progesterone receptor expression (ESR and PGR, respectively) and an absence of human epithelial growth factor receptor (ERBB2) amplification. Approximately 15–20% of breast malignancies are TNBC. Patients with TNBC often have an unfavorable prognosis. In addition, TNBC represents an important clinical challenge since it does not respond to hormone therapy. Methods In this work, we integrated high-throughput mRNA sequencing (RNA-Seq) data from normal and tumor tissues (obtained from The Cancer Genome Atlas, TCGA) and cell lines obtained through in-house sequencing or available from the Gene Expression Omnibus (GEO) to generate a unified list of differentially expressed (DE) genes. Methylome and proteomic data were integrated to our analysis to give further support to our findings. Genes that were overexpressed in TNBC were then curated to retain new potentially druggable targets based on in silico analysis. Knocking-down was used to assess gene importance for TNBC cell proliferation. Results Our pipeline analysis generated a list of 243 potential new targets for treating TNBC. We finally demonstrated that knock-down of Guanylate-Binding Protein 1 (GBP1 ), one of the candidate genes, selectively affected the growth of TNBC cell lines. Moreover, we showed that GBP1 expression was controlled by epidermal growth factor receptor (EGFR) in breast cancer cell lines. Conclusions We propose that GBP1 is a new potential druggable therapeutic target for treating TNBC with enhanced EGFR expression.

A 47-year-old male presented to the emergency department with 12 hours of nausea, vomiting, abdominal pain, and a widespread skin eruption with mottled, irregular, purpuric lesions with subsequent rapid decompensation. Laboratory analysis revealed thrombocytopenia, bandemia, elevated metamyelocytes, abnormal coagulation panel, decreased fibrinogen, elevated fibrin split products, renal dysfunction, bacterial rods, dohle bodies, and toxic granulation. Acute promyelocytic leukemia (APML) was confirmed via bone marrow biopsy, flow cytometry, and fluorescence in situ hybridization analysis. Disseminated intravascular coagulation (DIC) may be the initial presentation of APML. When treated promptly, APML can achieve high remission rates; however, conditions such as DIC continue to increase mortality requiring early recognition to improve survival rates.