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The ongoing increase in genomic surveillance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lineages remains a significant global challenge, mainly due to the incompatibility of genetic sequencing with the technological and economic capabilities of most countries worldwide. Additionally, the continued circulation and emergence of numerous lineages of the Omicron variant with some specific mutations represent a challenge in developing straightforward tests for their discrimination. In this study, we developed a multiplex assay based on reverse transcription loop-mediated isothermal amplification with lateral flow assay detection (RT-LAMP-LFA). This assay aims to enhance the accessibility of genomic surveillance and to deliver more precise epidemiological data to support public health decision-making. To demonstrate its utility, we used the assay as a proof of concept for discriminating between the SARS-CoV-2 Omicron lineages BA.1 and BA.2. When comparing the results of the new assay with the gold standard method of genetic sequencing in a panel of clinical samples, the multiplex RT-LAMP-LFA demonstrated excellent diagnostic performance. For BA.1 lineage detection, the assay achieved 100% sensitivity, 100% specificity, and 100% accuracy, while for BA.2, it showed 100% sensitivity, 95% specificity, and 96% accuracy. The overall simplicity of the method combined with advantages such as short analysis time (40 min), low cost (∼$15 per test) and adaptability to the point-of-care format make the multiplex RT-LAMP-LFA assay an important screening tool for inferring SARS-CoV-2 Omicron lineage positive samples, thus alleviating the high demand for sequencing and expanding genomic surveillance, even in remote locations.

Base editors are precise editing tools that employ deaminases to modify target DNA bases. The DYW-family of cytosine deaminases is structurally and phylogenetically distinct and might be harnessed for genome editing tools. We report a novel CRISPR/Cas9-cytosine base editor using SsdA, a DYW-like deaminase and bacterial toxin. A G103S mutation in SsdA enhances C-to-T editing efficiency while reducing its toxicity. Truncations result in an extraordinarily small enzyme. The SsdA-base editor efficiently converts C-to-T in rice and barley protoplasts and induces mutations in rice plants and mammalian cells. The engineered SsdA is a highly efficient genome editing tool., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Objective: Black individuals remain underrepresented in disordered eating research, despite evidence that both Black men and women present with disordered eating behaviors. Culturally-informed theoretical frameworks suggest that these behaviors may be linked to race-related sociocultural experiences, such as aspects of racial identity. While studies have focused on racial identity commitment, the association between attitudes toward one's racial identity and disordered eating remains underexplored. The present study examines whether positive attitudes toward one's Blackness and Black culture are associated with disordered eating., Method: In a cross-sectional online sample of Black men and women (N = 458), we measured self-reported attitudes toward Blackness (i.e., centrality and private regard) and disordered eating behaviors (i.e., purging, binge eating, excessive exercise, and drive for thinness)., Results: In pre-registered linear regression models, private regard was negatively associated with purging and binge eating. Across all models, centrality was not associated with disordered eating. On average, Black women reported greater drive for thinness whereas Black men reported higher excessive exercise scores., Discussion: This is the first study to demonstrate associations between racial attitudes and disordered eating among Black men and women. Our findings affirm unique correlates of disordered eating among Black people and suggest that positive attitudes toward one's Blackness and Black culture may be a protective factor against the development of disordered eating., (© 2024 The Author(s). International Journal of Eating Disorders published by Wiley Periodicals LLC.)

A hereditary component of breast (BC) and colorectal cancer (CRC) has been described in approximately one-third of these tumor types. BC patients have an increased risk of developing CRC as a second primary tumor and vice versa. Germline genomic variants (NextSeq550, Illumina) were investigated in 24 unrelated BC and/or CRC patients and 7 relatives from 3 index patients. Fifty-six pathogenic or likely pathogenic variants were identified in 19 of 24 patients. We detected single-nucleotide variants (SNVs) in CRC predisposition genes ( MLH1 and MUTYH ) and other promising candidates ( CDK5RAP3 , MAD1L1 , NOS3 , and POLM ). Eighteen patients presented SNVs or copy number variants (CNVs) in DNA damage repair genes. We also identified SNVs recently associated with BC or CRC predisposition ( PABPC1 , TYRO3 , MAP3K1 , SLC15A4 , and LAMA1 ). The PABPC1 c.1255C>T variant was detected in nine unrelated patients. Each patient presented at least one SNV/CNV in a candidate gene, and most had alterations in more than one gene, reinforcing a polygenic model for BC/CRC predisposition. A significant fraction of BC/CRC patients with a family history of these tumors harbored deleterious germline variants in DNA repair genes. Our findings can lead to strategies to improve the diagnosis, genetic counseling, and treatment of patients and their relatives.

Introduction: Overusing and misusing pesticides, including paraquat (PQ), have led to numerous environmental contamination complications. PQ is an emerging bio-accumulative contaminant that is present in environmental aqueous matrices. Adsorption techniques are part of a set of technologies applied in ecological remediation, known for their high effectiveness in removing aqueous PQ. A study of the PQ adsorption capacity of three cyclodextrin-silica nanocomposites (α-CDSi, β-CDSi, and γ-CDSi) from contaminated waters is presented in this paper., Methods: The cyclodextrin-silica nanocomposites were synthesized via an esterification reaction between the inorganic matrix and cyclodextrins (CDs) (α, β, and γ) and were characterized physicochemically by spectroscopic, thermal, and surface methods. Their PQ removal performance from contaminated aqueous media was studied under different experimental conditions., Results and Discussion: The results showed a fast adsorptive response in removal treatment studies over time. Adsorption capacities of 87.22, 57.17, and 77.27 mg.g -1 were found for α-CDSi, β-CDSi, and γ-CDSi, respectively, at only 30 min of treatment. Thermodynamic studies indicated spontaneous and exothermic adsorption processes. The removal assays responded mainly to physisorption mechanisms with contributions from chemisorption mechanisms. Spectroscopic assays showed a strong interaction of PQ with the adsorbents used. Innovative CDSi nanocomposites have proven to be highly efficient in applying aqueous PQ remediation, thus proving to be sustainable adsorbents of contaminants of emerging importance worldwide., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Baigorria, Carvalho, Alves Pinto and Fraceto.)

Objective: To analyze in silico the evidence of molecular mimicry between human beta-cell autoantigens and cow's milk proteins as a potential type 1 diabetes mellitus (T1DM) trigger., Materials and Methods: The in silico analysis was performed using bioinformatics tools to compare the amino acid sequences of cow's milk proteins (bovine serum albumin [BSA] and beta-lactoglobulin [BLG]) and human beta-cell autoantigens (glutamic acid decarboxylase-65 [GAD-65], insulin, and zinc transporter 8 [ZnT8]). The structural and functional characteristics of the proteins were analyzed to identify potential molecular mimicry mechanisms., Results: The results of the in silico analysis showed significant sequence similarity between BSA/BLG and GAD-65/human insulin/ZnT8, ranging from 19.64% to 27.27%. The cow's milk proteins evaluated shared structural features with the beta-cell antigens selected for comparison, indicating a potential for molecular mimicry between these proteins., Conclusion: The findings of this study provide further evidence for a potential role of cow's milk proteins in triggering T1DM. The in silico analysis suggests that molecular mimicry mechanisms between cow's milk proteins and human beta-cell antigens may contribute to the autoimmune response leading to T1DM., Competing Interests: Disclosure: no potential conflict of interest relevant to this article was reported.

Background: Amazonas was one of the most impacted Brazilian states by the COVID-19 pandemic. Mortality rates were high, and the health systems collapsed. It is important to identify possible intermediate reservoirs to avoid animal-to-human contamination. Several tropical fish are of commercial interest and are sold in large open-air markets in the region, representing a large economic and dietary importance., Objectives: This study aimed to verify if fish species of commercial importance, aerosols, and fish wastewater in local open-air markets, at a major capital city in the western Brazilian Amazon, are contaminated by SARS-CoV-2., Methods: 488 fish, 50 aerosol, and 45 wastewater samples were analyzed for the presence of SARS-CoV-2. The samples were subjected to extraction using the BIOGENE Viral DNA/RNA Extraction kit, and the molecular diagnosis was tested for SARS-CoV-2 using the Bio-Manguinhos SARS-CoV-2 (EDx) Molecular Kit., Results: It was not possible to detect the virus (Ct≤40, for Gene E) in these samples, however, in 181 samples of fish it was possible to detect the human RP gene (Ct≤35, for the RP Gene), indicating human contact. There was a high number of COVID-19 diagnoses in all city districts in which the samples were collected, showing that SARS-CoV-2 was circulating., Conclusion: This study indicates that fish of local commercial importance do not carry SARS-CoV-2 viral particles, despite circulation of SARS-CoV-2, and are not an important source of animal-to-human contamination. Despite these results, the human RP gene was found detectable in fish, air, and fish wastewater, showing that such places may carry human pathogens., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 de Sá Leitão et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: To evaluate the influence of chemotherapy on the prevalence of trismus in irradiated head and neck cancer patients., Material and Methods: This systematic review guided by PRISMA-2020 and registered in PROSPERO (CRD42021255377) screened 963 articles in 7 scientific-databases (PubMed, Lilacs, Livivo, Scopus, Embase, Web of Science, EBSCO) and 3 grey-literature databases (Open Grey, Google Scholar, ProQuest) and eight articles were included for qualitative synthesis, meta-analysis (combined odds ratio, inverse variance method plus random effects), heterogeneity analysis (I² and Tau²), one-of-out evaluation and publication bias analysis (Eggs' and Begg's tests) (RevMan®, p <0.05). The Newcastle-Ottawa Quality Assessment Scale Cohort Studies was used to assess the risk of bias (RoB). The classification assessment, development, and recommendations (GRADE) approach was used to assess the certainty of evidence., Results: The eight articles evaluated 1474 patients treated with chemoradiotherapy and 858 patients treated with radiotherapy. Five articles had low RoB, and three had high RoB. Chemoradiotherapy significantly ( p =0.0003) increased the prevalence of trismus (OR=2.55, 95% CI = 1.53-4.23) compared to radiotherapy, with significant ( p =0.010) but low heterogeneity (I²=59%;Tau²=0.29). There was no significant risk of publication bias, one-out analysis showed no significant difference between studies, and GRADE showed a moderate level of evidence. Trismus was directly associated to worse quality of life., Conclusions: The incidence of trismus increases when chemotherapy is combined with radiotherapy for head and neck cancer, which negatively impacts the quality of life. Key words: Radiotherapy, Chemoradiotherapy, Head and Neck Neoplasms, Trismus, Quality of Life., Competing Interests: Conflicts of interest The authors have no conflicts of interest., (Copyright: © 2024 Medicina Oral S.L.)

The disability of cells to react to insulin, causing glucose intolerance and hyperglycemia, is referred to as insulin resistance. This clinical condition, which has been well-researched in organs such as adipose tissue, muscle, and liver, has been linked to neurodegenerative diseases like Alzheimer's disease (AD) when it occurs in the brain., Objective: The authors aimed to gather data from the current literature on brain insulin resistance (BIR) and its likely repercussions on neurodegenerative disorders, more specifically AD, through a systematic review., Methods: A comprehensive search was conducted in multiple medical databases, including the Cochrane Central Register of Controlled Trials, EMBASE, Medical Literature Analysis and Retrieval System Online (Medline), and PubMed ® , employing the descriptors: "insulin resistance", "brain insulin resistance", "Alzheimer's disease", "neurodegeneration", and "cognition". The authors focused their search on English-language studies published between 2000 and 2023 that investigated the influence of BIR on neurodegenerative disorders or offered insights into BIR's underlying mechanisms. Seventeen studies that met the inclusion criteria were selected., Results: The results indicate that BIR is a phenomenon observed in a variety of neurodegenerative disorders, including AD. Studies suggest that impaired glucose utilization and uptake, reduced adenosine triphosphate (ATP) production, and synaptic plasticity changes caused by BIR are linked to cognitive problems. However, conflicting results were observed regarding the association between AD and BIR, with some studies suggesting no association., Conclusion: Based on the evaluated studies, it can be concluded that the association between AD and BIR remains inconclusive, and additional research is needed to elucidate this relationship., Competing Interests: Disclosure: The authors report no conflicts of interest.

Despite its rare frequency, a pleuroperitoneal communication is a well-documented complication for patients on peritoneal dialysis. It occurs in ~2% of continuous ambulatory peritoneal dialysis, with uncertain incidence for those on automated peritoneal dialysis. We report a case of a 30-year-old female patient with end-stage kidney disease with sudden dyspnea 2 days after starting automated peritoneal dialysis. Her chest x-ray revealed a significant pleural effusion on the right side. A thoracocentesis was performed, with a pleural glucose/plasma glucose of 1.08. Additionally, a computed tomography scan revealed a pleuroperitoneal communication upon dialysate infusion added with media contrast. A pleural-to-serum glucose gradient of greater than 50 mg/dL may indicate the diagnosis of a pleuroperitoneal communication in patients on peritoneal dialysis. Current literature also indicates that a pleural-to-serum glucose ratio above 1.0 may provide a more sensitive analysis. This case highlights the diagnosis process for this complication, with both laboratory and image findings corroborating the clinical hypotheses of a pleuroperitoneal communication in a patient on automated peritoneal dialysis., (© 2023 Asian Pacific Society of Nephrology.)

This study evaluated photobiomodulation therapy (PBMT) for treatment of trismus in patients undergoing radiotherapy for head and neck cancer (HNC). Sixteen patients, 10 men and 6 women, who had a mouth opening < 35 mm and underwent RT were included. The patients were evaluated daily before and after the PBMT application, measuring mouth opening and performing pain scores for the masticatory muscles using the visual analog scale (VAS). We used the infrared laser (~ 808 nm) extraorally, 0.1 W power, 3 J energy, 30 s (107 J/cm 2 ) per point, applied to temporalis anterior, masseter muscles, and temporomandibular joints (TMJ). An intraoral point was made in the trigonoretromolar region towards the medial pterygoid muscle. The mean mouth opening of the patients increased by more than 7 mm throughout the treatment. The pain scores on the initial days showed an immediate reduction after PBMT on the ipsilateral side in the muscles and TMJ. Throughout PBMT applications, there was a significant reduction in pain scores in all muscles and the TMJ. The radiation dose of all patients was above 40 Gy, which is the threshold dose for the risk of developing trismus. SPSS software was used and adopted a confidence of 95%. The Kolmogorov-Smirnov normality test, Wilcoxon test, and Spearman correlation were performed. PBMT controls muscular pain and reduced mouth opening limitation in HNC during radiotherapy. Further studies are needed to evaluate the preventive capacity of PBMT protocols for RT trismus-related HNC., (© 2023. The Author(s), under exclusive licence to Springer-Verlag London Ltd., part of Springer Nature.)

Bronchial anastomotic complications are a cause of grave concern for surgeons that perform lung transplantations. There are several risk factors that may lead to this complication, being inadequate surgical technique one of them, specifically regarding adequate exposure and manipulation of the bronchial stump and anastomosis. Here we report the use of Octopus™ Tissue Stabilizer as a mean to allow for a better exposure of the stump and facilitate a "no-touch" approach towards anastomosis. Systematic application of devices that facilitate the employment of the correct surgical techniques can have an effect in reducing the incidence of bronchial anastomotic complications.

Objectives  To analyze the lower limb strength in both untreated and surgically treated adolescent idiopathic scoliosis (AIS) patients and examine its correlation with the distance covered in a six-minute walking test (6MWT). Methods  A total of 88 participants (n = 30 pre-surgery AIS patients, n = 30 post-surgical AIS patients, and n = 28 control) underwent a 6MWT and a muscle strength assessment. The lower limb strength was measured at the knee joint using the knee extension (KE) and knee flexion (KF) peak torque (PT) measurements. Results  The control group covered a greater distance in the TC6 compared to both the pre-surgical (534 ± 67 m) and post-surgical (541 ± 69 m) groups, with a distance of 612 ± 70 m (p < 0.001). No differences were observed in KE PT (pre: 2.1 ± 0.63, post: 2.1 ± 0.7, control: 2.2 ± 0.7 Nm.kg -1 , p = 0.67) or KF PT (pre: 1.0 ± 0.3, post: 1.1 ± 0.3, control: 1.1 ± 0.5 Nm.kg -1 , p = 0.46). A moderate positive correlation was observed between KE PT and 6MWT distance (r = 0.53, p < 0.001), as well as a low positive correlation for KF PT (r = 0.37, p = 0.003) with 6MWT distance. Conclusion  This study highlights the importance of lower limb maximal strength in the functionality of AIS patients. Our findings suggest that exercise programs aimed at enhancing lower limb strength, especially the KE, could improve the walking capacity of AIS patients. These results provide useful information for designing purposeful exercise programs for AIS patients with walking deficits., Competing Interests: Conflito de Interesses Os autores declaram não haver conflito de interesses., (Sociedade Brasileira de Ortopedia e Traumatologia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. ( https://creativecommons.org/licenses/by-nc-nd/4.0/ ).)

Background: Patients with colorectal metastatic disease have a poor prognosis, limited therapeutic options, and frequent development of resistance. Strategies based on tumor-derived organoids are a powerful tool to assess drug sensitivity at an individual level and to suggest new treatment options or re-challenge. Here, we evaluated the method's feasibility and clinical outcome as applied to patients with no satisfactory treatment options., Methods: In this phase 2, single-center, open-label, non-comparative study (ClinicalTrials.gov, register NCT03251612), we enrolled 90 patients with metastatic colorectal cancer following progression on or after standard therapy. Participants were 18 years or older with an Eastern Cooperative Oncology Group performance status of 0-2, adequate organ function, and metastasis available for biopsy. Biopsies from the metastatic site were cultured using organoids model. Sensitivity testing was performed with a panel of drugs with proven activity in phase II or III trials. At the discretion of the investigator considering toxicity, the drug with the highest relative activity was offered. The primary endpoint was the proportion of patients alive without disease progression at two months per local assessment., Results: Biopsies available from 82 to 90 patients were processed for cell culture, of which 44 successfully generated organoids with at least one treatment suggested. The precision cohort of 34 patients started treatment and the primary endpoint, progression-free survival (PFS) at two months was met in 17 patients (50%, 95% CI 32-68), exceeding the pre-defined level (14 of 45; 31%). The median PFS was 67 days (95% CI 51-108), and the median overall survival was 189 days (95% CI 103-277)., Conclusions: Patient-derived organoids and in-vitro sensitivity testing were feasible in a cohort of metastatic colorectal cancer. The primary endpoint was met, as half of the patients were without progression at two months. Cancer patients may benefit from functional testing using tumor-derived organoids., Trial Registration: ClinicalTrials.gov, register NCT03251612., (© 2023. The Author(s).)

Objective: To define the mathematical relationship between fructosamine levels and average glucose values., Subjects and Methods: The study comprised laboratory data of 1,227 patients with type 1 or 2 diabetes mellitus. Fructosamine levels measured at the end of a 3-week period were compared against the average blood glucose levels of the previous 3 weeks. Average glucose levels were determined by the weighted average of the daily fasting capillary glucose results performed during the study period, and the plasma glucose measured in the same sample collected for fructosamine measurement., Results: In total, 9,450 glucose measurements were performed. Linear regression analysis between fructosamine levels and average glucose levels showed that for each 1.0 μmol/L increase in fructosamine level there was a 0.5 mg/dL increase in average glucose level, as estimated by the equation Mean glucose level = (0.5157 × Fructosamine) - 20 . The coefficient of determination (r2 = 0.353492, p < 0.006881) allowed the calculation of the estimated average glucose based on fructosamine level., Conclusion: Our study demonstrated a linear correlation between fructosamine level and mean blood glucose level, suggesting that fructosamine levels can be a proxy for the average glucose level in assessing the metabolic control of patients with diabetes.

The rapid proliferation of renewable energy communities/ecosystems is an indication of their potential contribution to the ongoing energy transition. A common characteristic of these ecosystems is their complex composition, which often involves the interaction of multiple actors. Currently, the notions of "networking", "collaboration", "coordination", and "cooperation", although having different meanings, are often loosely used to describe these interactions, which creates a sense of ambiguity and confusion. To better characterize the nature of interactions in current and emerging ecosystems, this article uses the systematic literature review method to analyse 34 emerging cases. The objective is threefold (a) to study the interactions and engagements between the involved actors, aiming at identifying elements of collaboration. (b) Identify the adopted technological enablers, and (c) ascertain how the composition and functions of these ecosystems compare to virtual power plants. The outcome revealed that the interactions between the members of these ecosystems can be described as cooperation and not necessarily as collaboration, except in a few cases. Regarding technological enablers, a vast panoply of technologies, such as IoT devices, smart meters, intelligent software agents, peer-to-peer networks, distributed ledger systems/blockchain technology (including smart contracts, blockchain as a platform service, and cryptocurrencies) were found. In comparison with virtual power plants, these ecosystems have similar composition, thus, having multiple actors, comprised of decentralized and heterogeneous technologies, and are formed by aggregating various distributed energy resources. They are also supported by ICT and are characterized by the simultaneous flow of information and energy., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s).)

Background: During oral rehabilitation, dental implants in the posterior maxilla can penetrate the maxillary sinus. The aim was to evaluate the presence of maxillary sinus abnormalities in patients with dental implants in the posterior maxillary region using cone-beam computed tomography (CBCT) images., Materials and Methods: This was a retrospective cross-sectional study, and CBCT scans of 199 patients (459 dental implants) were evaluated. Implants were assessed according to their relative location to the maxillary sinus floor (up to 2 mm from the maxillary sinus cortex, within 2 mm to intimate contact with the maxillary sinus cortex, apical third inside the maxillary sinus, two-thirds or more inside the maxillary sinus) and bone-fixation tissue (Alveolar ridge or Bone graft). Maxillary sinus abnormalities were classified. Kappa and Weighted Kappa and the Kruskal-Wallis test were applied., Results: A higher prevalence of mucosal thickening and non-specific opacification were observed in implants located within 2 mm to intimate contact with the cortex of the maxillary sinus floor. Of the 66 implants with apical thirds located inside the maxillary sinus, 31 (46.7%) were associated with sinus abnormalities and of all implants ( n = 5) with two-thirds or more located inside the maxillary sinus, all of these were associated with sinus abnormalities. No association was observed in relation to implant bone-fixation tissue., Conclusions: This study found a significant association between dental implant placement near or within the sinus and sinus abnormalities, mainly mucosal thickening and non-specific opacification.

In recent years, the manufacturing sector is going through a major transformation, as reflected in the concept of Industry 4.0 and digital transformation. The urge for such transformation is intensified when we consider the growing societal demands for sustainability. The notion of sustainable manufacturing has emerged as a result of this trend. Additionally, industries and the whole society face the challenges of an increasing number of disruptive events, either natural or human-caused, that can severely affect the normal operation of systems. Furthermore, the growing interconnectivity between organizations, people, and physical systems, supported by recent developments in information and communication technologies, highlights the important role that collaborative networks can play in the digital transformation processes. As such, this article analyses potential synergies between the areas of sustainable and resilient manufacturing and collaborative networks. The work also discusses how the responsibility for the various facets of sustainability can be distributed among the multiple entities involved in manufacturing. The study is based on a literature survey, complemented with the experience gained from various research projects and related initiatives in the area, and is organized according to various dimensions of Industry 4.0. A brief review of proposed approaches and indicators for measuring sustainability from the networked manufacturing perspective is also included. Finally, a set of key research challenges are identified to complement strategic research agendas in manufacturing., (© The Author(s) 2022.)

Background: Serous ovarian carcinoma is the most frequent histological subgroup of ovarian cancer and the leading cause of death among gynecologic tumors. The tumor microenvironment and cancer-associated fibroblasts (CAFs) have a critical role in the origin and progression of cancer. We comprehensively characterized the crosstalk between CAFs and ovarian cancer cells from malignant fluids to identify specific ligands and receptors mediating intercellular communications and disrupted pathways related to prognosis and therapy response., Methods: Malignant fluids of serous ovarian cancer, including tumor-derived organoids, CAFs-enriched (eCAFs), and malignant effusion cells (no cultured) paired with normal ovarian tissues, were explored by RNA-sequencing. These data were integrated with single-cell RNA-sequencing data of ascites from ovarian cancer patients. The most relevant ligand and receptor interactions were used to identify differentially expressed genes with prognostic values in ovarian cancer., Results: CAF ligands and epithelial cancer cell receptors were enriched for PI3K-AKT, focal adhesion, and epithelial-mesenchymal transition signaling pathways. Collagens, MIF, MDK, APP, and laminin were detected as the most significant signaling, and the top ligand-receptor interactions THBS2/THBS3 (CAFs)-CD47 (cancer cells), MDK (CAFs)-NCL/SDC2/SDC4 (cancer cells) as potential therapeutic targets. Interestingly, 34 genes encoding receptors and ligands of the PI3K pathway were associated with the outcome, response to treatment, and overall survival in ovarian cancer. Up-regulated genes from this list consistently predicted a worse overall survival (hazard ratio &gt; 1.0 and log-rank P &lt; 0.05) in two independent validation cohorts., Conclusions: This study describes critical signaling pathways, ligands, and receptors involved in the communication between CAFs and cancer cells that have prognostic and therapeutic significance in ovarian cancer. Video abstract., (© 2022. The Author(s).)

Periods of soil water stress have been recurrent in the Cerrado region and have become a growing concern for Brazilian tropical pasture areas. Thus, the search for forage grasses more tolerant to water stress has intensified recently in order to promote more sustainable livestock. In a greenhouse experiment, the degree of water stress tolerance of nine tropical forage grass cultivars was studied under different soil water regimes. The investigation followed a 9 × 3 factorial design in four randomized blocks. Nine cultivars from five species of perennial forage grasses were tested: Urochloa brizantha ('BRS Piatã', 'Marandu', and 'Xaraés'), Panicum maximum ('Aruana', 'Mombaça', and 'Tanzânia'), Pennisetum glaucum ('ADR 300'), Urochloa ruziziensis ('Comum'), and Paspalum atratum ('Pojuca'). These cultivars were grown in pots under three soil water regimes (high soil water regime-HSW (non-stressful condition), middle soil water regime-MSW (moderate water stress), and low soil water regime-LSW (severe water stress)). Plants were exposed to soil water stress for 25 days during the tillering and stalk elongation phases. Twelve tolerance indices, including tolerance index (TOL), mean production (MP), yield stability index (YSI), drought resistance index (DI), stress tolerance index (STI), geometric mean production (GMP), yield index (YI), modified stress tolerance (k 1 STI and k 2 STI), stress susceptibility percentage index (SSPI), abiotic tolerance index (ATI), and harmonic mean (HM), were calculated based on shoot biomass production under non-stressful (Y P ) and stressful (Y S ) conditions. Soil water stress decreased leaf area, plant height, tillering capacity, root volume, and shoot and root dry matter production in most cultivars, with varying degrees of reduction among tropical forage grasses. Based on shoot biomass production under controlled greenhouse conditions, the most water-stress-tolerant cultivars were P. maximum cv. Mombaça and cv. Tanzânia under the MSW regime and P. maximum cv. Aruana and cv. Mombaça under the LSW regime. P. maximum cv. Mombaça has greater adaptability and stability of shoot biomass production when grown under greenhouse conditions and subjected to soil water stress. Therefore, this forage grass should be tested under field conditions to confirm its forage production potential for cultivation in tropical regions with the occurrence of water stress. The MP, DI, STI, GMP, YI, k 2 STI, and HM tolerance indices were the most suitable for identifying forage grass cultivars with greater water stress tolerance and a high potential for shoot biomass production under LSW regime.

Implementation of resource recovery technologies is becoming increasingly important, as humans are exhausting the world's natural resources. Recovering nutrients and water from wastewater treatment systems will play an important role in changing the current trends towards a circular economy. However, guidance is still needed to determine the most appropriate way to do this. In this study two decision-support tools, sanitation planning software (Santiago) and life cycle assessment (LCA), were applied to identify appropriate technologies and their environmental impacts. As a case study, current and alternative scenarios for a wastewater treatment plant (WWTP) in Campo Grande, west-central Brazil, were used. Among 12 scenarios provided by Santiago for efficient nutrient recovery, eight were selected for further assessment. The current WWTP system (UASB reactors) resulted in the highest negative impacts in two of nine assessment categories (freshwater and marine eutrophication), due to nutrient discharge to water. A source separation scenario with urine stored in a urine bank and co-composting of feces showed best overall performance. Electricity consumption played a crucial role for impacts in several categories, while water consumption was not significantly affected by choice of toilet. One Santiago scenario matched the most appropriate scenario with the best environmental performance, but the other seven scenarios were not as beneficial, indicating a need for some adjustments in the software. These results highlight the importance of performing LCA to compare alternative scenarios, even when using a tool designed to identify locally appropriate technologies. The results also indicate that the current wastewater treatment system has reasonable environmental performance, but could be improved if measures were taken to recover energy and reuse water., Competing Interests: Declaration of competing interest Priscila de Morais Lima reports financial support was provided by Swedish Research Council., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Valvular heart disease is an important contributor to cardiovascular morbidity and mortality and remains underdiagnosed. Deep learning analysis of electrocardiography (ECG) may be useful in detecting aortic stenosis (AS), aortic regurgitation (AR), and mitral regurgitation (MR)., Objectives: This study aimed to develop ECG deep learning algorithms to identify moderate or severe AS, AR, and MR alone and in combination., Methods: A total of 77,163 patients undergoing ECG within 1 year before echocardiography from 2005-2021 were identified and split into train (n = 43,165), validation (n = 12,950), and test sets (n = 21,048; 7.8% with any of AS, AR, or MR). Model performance was assessed using area under the receiver-operating characteristic (AU-ROC) and precision-recall curves. Outside validation was conducted on an independent data set. Test accuracy was modeled using different disease prevalence levels to simulate screening efficacy using the deep learning model., Results: The deep learning algorithm model accuracy was as follows: AS (AU-ROC: 0.88), AR (AU-ROC: 0.77), MR (AU-ROC: 0.83), and any of AS, AR, or MR (AU-ROC: 0.84; sensitivity 78%, specificity 73%) with similar accuracy in external validation. In screening program modeling, test characteristics were dependent on underlying prevalence and selected sensitivity levels. At a prevalence of 7.8%, the positive and negative predictive values were 20% and 97.6%, respectively., Conclusions: Deep learning analysis of the ECG can accurately detect AS, AR, and MR in this multicenter cohort and may serve as the basis for the development of a valvular heart disease screening program., Competing Interests: Funding Support and Author Disclosures This study was supported by a National Institutes of Health Institutional Training Grant (2T32HL007854-21) and National Institutes of Health/National Heart, Lung, and Blood Institute Award (R01HL148248) (to Dr Elias and Dr Perotte’s institution). Dr Elias has received research grant support provided to his institution from Pfizer, Eidos Therapeutics, Google, and Edwards LifeSciences; and is an inventor on a pending patent related to the ValveNet algorithm described in this paper. Dr Poterucha owns stock in Abbott Laboratories and Baxter International with research support provided to his institution from the Amyloidosis Foundation, Eidos Therapeutics, Pfizer, Edwards Lifesciences, and the Glorney-Raisbeck Fellowship Award from the New York Academy of Medicine; and is an inventor on a pending patent related to the ValveNet algorithm described in this paper. Dr Hahn has received speaker fees from Abbott Structural, Edwards Lifesciences, and Philips Healthcare; has institutional consulting contracts with Abbott Structural, Boston Scientific, Edwards Lifesciences, and Gore and Associates; has equity with Navigate; and is Chief Scientific Officer for the Echocardiography Core Laboratory at the Cardiovascular Research Foundation for multiple industry-sponsored trials, for which she receives no direct industry compensation. Dr Tison has received research grants from General Electric, Janssen Pharmaceuticals, and Myokardia; has received personal fees from Myokardia Digital Health as an advisory group member; and has served as an unpaid advisor for Cardiogram. Dr Kodali has served as a consultant for or received honoraria from Admedus, Meril Lifesciences, JenaValve, and Abbott Vascular; has served on the scientific advisory board for and owns equity in Dura Biotech, MicroInterventional Devices, Thubrikar Aortic Valve, Supira, and Admedus; and has received institutional funding to Columbia University and/or the Cardiovascular Research Foundation from Edwards Lifesciences, Medtronic, Abbott Vascular, Boston Scientific, and JenaValve. Dr Khalique has served as a consultant for Boston Scientific, Edwards Lifesciences, and Abbott Structural. Dr Cohen has received research grant support and consulting income from Edwards Lifesciences, Abbott, Boston Scientific, and Medtronic. Dr Einstein has received speaker fees from Ionetix; has received consulting fees from W. L. Gore and Associates; has received authorship fees from Wolters Kluwer Healthcare–UpToDate; and his institution has grants/grants pending from Attralus, Canon Medical Systems, Eidos Therapeutics, GE Healthcare, Pfizer, Roche Medical Systems, W. L. Gore and Associates, and XyloCor Therapeutics. Dr Nazif has served as a consultant for or received honoraria from Edwards Lifesciences, Medtronic, Venus Medtech, and Boston Scientific. Dr Leon has received institutional grants for clinical research from Abbott, Boston Scientific, Edwards, JenaValve, and Medtronic; has received stock options (equity) for advisory board participation in Valve Medical, Picardia, and Venus MedTech; and is an inventor on a pending patent related to the ValveNet algorithm described in this paper. Dr Perotte is Chief Scientific and Innovation Officer at Spiden AG; and is an inventor on a pending patent related to the ValveNet algorithm described in this paper. All other authors have reported that they have no relationships relevant to the contents of this paper to disclose., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

High-throughput DNA sequencing has allowed for the identification of genomic alterations and their impact on tumor development, progression, and therapeutic responses. In PSCC, for which the incidence has progressively increased worldwide, there are still limited data on the molecular mechanisms involved in the disease pathogenesis. In this study, we characterized the mutational signature of 30 human papillomavirus (HPV)-associated PSCC cases from Latin Americans, using whole-exome sequencing. Copy number variations (CNVs) were also identified and compared to previous array-generated data. Enrichment analyses were performed to reveal disrupted pathways and to identify alterations mapped to HPV integration sites (HPVis) and miRNA-mRNA hybridization regions. Among the most frequently mutated genes were NOTCH1 , TERT , TTN , FAT1 , TP53 , CDKN2A , RYR2 , CASP8 , FBXW7 , HMCN2 , and ITGA8 . Of note, 92% of these altered genes were localized at HPVis. We also found mutations in ten novel genes ( KMT2C , SMARCA4 , PTPRB , AJUBA , CR1 , KMT2D , NBEA , FAM135B , GTF2I , and CIC ), thus increasing our understanding of the potential HPV-disrupted pathways. Therefore, our study reveals innovative targets with potential therapeutic benefits for HPV-associated PSCCs. The CNV analysis by sequencing (CNV-seq) revealed five cancer-associated genes as the most frequent with gains ( NOTCH1 , MYC , NUMA1 , PLAG1 , and RAD21 ), while 30% of the tumors showed SMARCA4 with loss. Additionally, four cancer-associated genes ( CARD11 , CSMD3 , KDR , and TLX3 ) carried untranslated regions (UTRs) variants, which may impact gene regulation by affecting the miRNAs hybridization regions. Altogether, these data contribute to the characterization of the mutational spectrum and its impact on cellular signaling pathways in PSCC, thus reinforcing the pivotal role of HPV infection in the molecular pathogenesis of these tumors.

Objective: To analyze the temporal trend of the incidence and mortality rate for prostate, breast, colorectal, lung, cervical, stomach and laryngeal cancer among residents in the city of Cuiabá between 2008 and 2016., Methods: Time series study with data from the Mortality Information System and the Population-Based Cancer Registry. Stratified by sex, the proportional distribution of new cases and deaths by age group and the cancer incidence and mortality rates standardized by the world population were calculated. Linear regression was used and the annual percentage change (APC) was estimated., Results: In males, most new cases and deaths, for the main types of cancer, occurred among those aged 50 years or older, and the incidence rate of prostate cancer showed a tendency to decrease in the period (APC=-4.33%). For females, the proportion of new cases and deaths, due to breast and cervical cancer, were more frequent among women aged 50 years or younger, and lung, stomach and colorectal cancer among women aged 50 years or older. The incidence rate of breast cancer showed an increasing trend (APC=3.60%). For both sexes, the mortality rate remained stable., Conclusion: The incidence rate trend varied between sexes, an increase was observed for breast cancer among women and a reduction for prostate cancer among men. The mortality rate for the main types of cancer was stable.

Background: Tracheal diverticulum (TD) is a rare entity in clinical practice, accidentally found by imaging methods. It is a potential factor for the development of chronic respiratory infections, contributing to the progression of preexisting lung diseases and putting the success of lung transplantation at risk., Objectives: This paper reports 2 cases of TD with atypical clinical presentation in post-lung transplant patients with recurrent infections and aims to present the importance of this differential diagnosis., Discussion: Case 1: A 30-year-old man with terminal lung disease underwent bilateral lung transplantation with a satisfactory postoperative period. He presented with TD as a focus of recurrent infection associated with persistent hemoptysis. Indicated for surgical resection (cervicotomy with resection of tracheal diverticulum), without complications. He evolved uneventfully in the postoperative period and was clinically stable at follow-up. Case 2: A 57-year-old woman with hypersensitivity pneumonia associated with secondary pulmonary arterial hypertension and bronchiectasis underwent bilateral lung transplantation without complications. She presented with TD as a focus of infection associated with esophageal symptoms. Indicated for the surgical approach (cervicotomy with resection of tracheal diverticulum), she was asymptomatic at follow-up., Conclusion: Conservative treatment is recommended in elderly and asymptomatic patients. Surgical resection is based on the recurrence of symptoms and failure of clinical therapy and is the preferred approach for reported cases., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: Guillian Barré syndrome (GBS) is an acute autoimmune polyradiculoneuropathy often associated with previous exposure to infectious agents., Methods: A clinical cohort of 41 patients with GBS admitted to the Base Hospital Institute of the Federal District between May 2017 and April 2019 was followed up for 1 year. Serological tests for arbovirus detection and amplification of nucleic acids using polymerase chain reaction for zika virus (ZIKV), dengue virus (DENV), and chikungunya virus (CHIKV) were performed., Results: The cohort consisted of 61% men with a median age of 40 years, and 83% had GBS-triggering events. A total of 54% had Grade 4 disability, 17% had Grade 3, 12% had Grade 2, 10% had Grade 5, and 7% had Grade 1. The classic form occurred in 83% of patients. Nerve conduction evaluations revealed acute demyelinating inflammatory polyneuropathy (51%), acute motor axonal neuropathy (17%), acute sensory-motor neuropathy (15%), and indeterminate forms (17%). Four patients were seropositive for DENV. There was no laboratory detection of ZIKV or CHIKV infection. Ninety percent of patients received human immunoglobulin. Intensive care unit admission occurred in 17.1% of the patients, and mechanical ventilation was used in 14.6%. One patient died of Bickerstaff's encephalitis. Most patients showed an improvement in disability at 10 weeks of follow-up., Conclusions: GBS in the Federal District showed a variable clinical spectrum, and it was possible to detect recent exposure to DENV.

Genetic and epigenetic changes contribute to intratumor heterogeneity and chemotherapy resistance in several tumor types. LncRNAs have been implicated, directly or indirectly, in the epigenetic regulation of gene expression. We investigated lncRNAs that potentially mediate carboplatin-resistance of cell subpopulations, influencing the progression of ovarian cancer (OC). Four carboplatin-sensitive OC cell lines (IGROV1, OVCAR3, OVCAR4, and OVCAR5), their derivative resistant cells, and two inherently carboplatin-resistant cell lines (OVCAR8 and Ovc316) were subjected to RNA sequencing and global DNA methylation analysis. Integrative and cross-validation analyses were performed using external (The Cancer Genome Atlas, TCGA dataset, n = 111 OC samples) and internal datasets ( n = 39 OC samples) to identify lncRNA candidates. A total of 4255 differentially expressed genes (DEGs) and 14529 differentially methylated CpG positions (DMPs) were identified comparing sensitive and resistant OC cell lines. The comparison of DEGs between OC cell lines and TCGA-OC dataset revealed 570 genes, including 50 lncRNAs, associated with carboplatin resistance. Eleven lncRNAs showed DMPs, including the SNHG12 . Knockdown of SNHG12 in Ovc316 and OVCAR8 cells increased their sensitivity to carboplatin. The results suggest that the lncRNA SNHG12 contributes to carboplatin resistance in OC and is a potential therapeutic target. We demonstrated that SNHG12 is functionally related to epigenetic mechanisms.

Background: Lung transplantation (LTx) has been discussed as an option for treating irreversible lung fibrosis post-coronavirus disease 2019 (COVID-19), in selected cases., Objectives: To report on the initial experience and management of end-stage lung disease due to COVID-19 at a national center reference in Brazil., Design and Setting: Cohort study conducted at a national reference center for lung transplantation., Methods: Medical charts were reviewed regarding patients' demographics and pre-COVID-19 characteristics, post-LTx due to COVID-19., Results: Between March 2020 and September 2021, there were 33 cases of LTx. During this period, we evaluated 11 cases of severe COVID-19-related acute respiratory distress syndrome (ARDS) that were potentially candidates for LTx. Among these, LTx was only indicated for three patients (9.1%). All of these patients were on venovenous extracorporeal membrane oxygenation (ECMO), and the procedure that they underwent was central venoarterial ECMO. All three patients were still alive after the first 30 postoperative days. However, patient #1 and patient #2 subsequently died due to fungal sepsis on the 47th and 52nd postoperative days, respectively. Patient #3 was discharged on the 30th postoperative day., Conclusions: LTx is feasible among these complex patients. Survival over the first 30 days was 100%, and this favors surgical feasibility. Nonetheless, these were critically ill patients.

Carbon monoxide (CO), nitric oxide (NO) and hydrogen sulfide (H 2 S) are gasotransmitters known for their roles in plant response to (a)biotic stresses. The crosstalk between these gasotransmitters and potassium ions (K + ) has received considerable attention in recent years, particularly due to the dual role of K + as an essential mineral nutrient and a promoter of plant tolerance to abiotic stress. This review brings together what it is known about the interplay among NO, CO, H 2 S and K + in plants with focus on the response to high salinity. Some findings obtained for plants under water deficit and metal stress are also presented and discussed since both abiotic stresses share similarities with salt stress. The molecular targets of the gasotransmitters NO, CO and H 2 S in root and guard cells that drive plant tolerance to salt stress are highlighted as well., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

Introduction: Supplementation with high doses of vitamin D, known as mega-dose or "stoss therapy," refers to administering high doses of vitamin D by oral or intramuscular route in short periods of time. In this sense, conducting a review to organize this knowledge in a single article will generate a helpful instrument for researchers working in this area and for health professionals who use this therapeutic tool., Objective: To carry out a literature review on safety and efficacy (normalization of serum vitamin D level, and changes in the clinical picture) of vitamin D mega-dose use., Methodology: This is a systematic review of the literature searching data through the electronic banks of PubMed, Scielo, and LILACS, using the following keywords: "vitamin D," "mega-dose," "stoss therapy," "cholecalciferol," in different combinations. CONSORT questionnaire was used to assess the quality of the included studies., Results: Of the 59 articles screened for this review, 10 were included in the review, studying patients with rickets, osteoporosis, and critically ill patients. Two studies compared the exact dosage of vitamin D by different routes of administration, and three studies compared different doses by the same route. All others studied vitamin D mega-dose versus placebo. Adverse effects were observed through the presence of hypercalcemia/hypercalciuria. Serum vitamin D levels were normalized between 70 and 100% of patients, and adverse effects ranged between 1.9 and 18.5%., Conclusion: The study demonstrated that vitamin D mega-dose therapy is effective in normalizing serum vitamin levels, and the toxicity assessed through adverse effects was low, with no expressive clinical significance. Despite this, there is still a need for further studies in the area to confirm the results found., Competing Interests: Declaration of competing interest None., (Copyright © 2021 European Society for Clinical Nutrition and Metabolism. Published by Elsevier Ltd. All rights reserved.)

Introduction: Guillain-Barre Syndrome (GBS) is an acute immune-mediated polyneuropathy that compromises the peripheral and cranial nerves. It is characterized by rapid-onset paresthesia accompanied by progressive weakness in the lower extremities followed by symmetric ascending paralysis., Methodology: assessment of sensitivity to detect GBS between March 2017 and May 2019 in a public referral hospital, using the capture-recapture method based on the Chapman estimator and comparing three GBS data sources: the hospital-based sentinel surveillance system (VSBH), Human Immunoglobulin Dispensing Records System (RDIH), and Hospital Information System (SIH)., Results: A total of 259 possible cases were identified (captured). Of these, 58 were confirmed and most resided in the Federal District. The VSBH showed the greatest sensitivity in case identification. The temporal distribution of cases showed periods with no cases identified, and more were registered during the rainy season from October to May, when high temperatures also occur., Conclusions: Increased circulation of arboviruses and gastrointestinal infections during the rainy season may explain the greater concentration of GBS cases. It is important to note that one-third of the cases identified in the different data sources do not converge, demonstrating that no single surveillance system is 100% effective. The severity and possible increase in cases related to GBS demonstrates the need for an improved surveillance system capable of monitoring and following-up cases involving neurological syndromes, regardless of the event preceding infection., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2021 Ana Flavia de Morais Oliveira, Luciana Guerra Gallo, Mabia Milhomem Bastos, Amanda Amaral Abrahão, Paulo Rufalco-Moutinho, Juliane Maria Alves Siqueira Malta, Klauss Kleydmann Sabino Garcia, Luiza Morais de Matos, Wildo Navegantes de Araujo.)

Rectal cancer is a common disease with high mortality rates and limited therapeutic options. Here we combined the gene expression signatures of rectal cancer patients with the reverse drug-induced gene-expression profiles to identify drug repositioning candidates for cancer therapy. Among the predicted repurposable drugs, topoisomerase II inhibitors (doxorubicin, teniposide, idarubicin, mitoxantrone, and epirubicin) presented a high potential to reverse rectal cancer gene expression signatures. We showed that these drugs effectively reduced the growth of colorectal cancer cell lines closely representing rectal cancer signatures. We also found a clear correlation between topoisomerase 2A ( TOP2A) gene copy number or expression levels with the sensitivity to topoisomerase II inhibitors. Furthermore, CRISPR-Cas9 and shRNA screenings confirmed that loss-of-function of the TOP2A has the highest efficacy in reducing cellular proliferation. Finally, we observed significant TOP2A copy number gains and increased expression in independent cohorts of rectal cancer patients. These findings can be translated into clinical practice to evaluate TOP2A status for targeted and personalized therapies based on topoisomerase II inhibitors in rectal cancer patients.

Despite contemporary research efforts, the prognosis of penile squamous cell carcinoma (PeSCC) has not significantly improved over the past decade. Despite frequently encountered patient-related delayed medical consultations impairing outcomes, several other aspects contribute to the lack of advancement in the treatment of this condition. One essential reason is that translational research, a prerequisite for the clinically successful disease management, is still at an early stage in PeSCC as compared to many other malignancies. Preclinical experimental models are indispensable for the evaluation of tumor biology and identification of genomic alterations. However, since neither commercial PeSCC cell lines are available nor xenograft models sustainably established, such analyses are challenging in this field of research. In addition, systemic therapies are less effective and toxic without decisive breakthroughs over recent years. Current systemic management of PeSCC is based on protocols that have been investigated in small series of only up to 30 patients. Thus, there is an unmet medical need for new approaches necessitating research efforts to develop more efficacious systemic strategies. This review aims to highlight the current state of knowledge in the molecular alterations involved in the etiology and ensuing steps for cancer progression, existing preclinical models of translational research, clinically relevant systemic protocols, and ongoing clinical trials., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/tau-20-945). The series “Management of Advanced Genitourinary Malignancies” was commissioned by the editorial office without any funding or sponsorship. Dr. PES reports other from NCCN Bladder and Penile Cancer Panel, other from Global Society of Rare GU Tumors, during the conduct of the study. The authors have no other conflicts of interest to declare., (2021 Translational Andrology and Urology. All rights reserved.)

Paracoccidioidomycosis (PCM) is a systemic disease caused by Paracoccidioides spp. PCM is endemic in Latin America and most cases are registered in Brazil. This mycosis affects mainly the lungs, but can also spread to other tissues and organs, including the liver. Several approaches have been investigated to improve treatment effectiveness and protection against the disease. Extracellular vesicles (EVs) are good antigen delivery vehicles. The present work aims to investigate the use of EVs derived from Paracoccidioides brasiliensis as an immunization tool in a murine model of PCM. For this, male C57BL/6 were immunized with two doses of EVs plus adjuvant and then infected with P. brasiliensis . EV immunization induced IgM and IgG in vivo and cytokine production by splenocytes ex vivo. Further, immunization with EVs had a positive effect on mice infected with P. brasiliensis , as it induced activated T lymphocytes and NKT cell mobilization to the infected lungs, improved production of proinflammatory cytokines and the histopathological profile, and reduced fungal burden. Therefore, the present study shows a new role for P. brasiliensis EVs in the presence of adjuvant as modulators of the host immune system, suggesting their utility as immunizing agents.

The number of people living with dementia in the world is rising at an unprecedented rate, and no country will be spared. Furthermore, neither decisive treatment nor effective medicines have yet become effective. One potential alternative to this emerging challenge is utilizing supportive technologies and services that not only assist people with dementia to do their daily activities safely and independently, but also reduce the overwhelming pressure on their caregivers. Thus, for this study, a systematic literature review is conducted in an attempt to gain an overview of the latest findings in this field of study and to address some commercially available supportive technologies and services that have potential application for people living with dementia. To this end, 30 potential supportive technologies and 15 active supportive services are identified from the literature and related websites. The technologies and services are classified into different classes and subclasses (according to their functionalities, capabilities, and features) aiming to facilitate their understanding and evaluation. The results of this work are aimed as a base for designing, integrating, developing, adapting, and customizing potential multimodal solutions for the specific needs of vulnerable people of our societies, such as those who suffer from different degrees of dementia.

Objectives: To evaluate four different teaching methodologies in the radiographic diagnosis of proximal carious lesions and in the students' perception of these methodologies., Methods: 71 undergraduate dental students were subdivided into 4 groups according to the teaching methodologies used (traditional, hybrid, e-learning and problem-based learning). All methods were applied by two properly trained researchers. Initially, students completed a pre-methodology test (index test). After applying the teaching methodologies, the students were submitted to a post-methodology test containing periapical radiographs for evaluation of proximal carious lesions. Then, the students answered a questionnaire to verify their perception of the different teaching methodologies. Statistical analysis was done., Results: There were no relationship between the results of pre- and post-methodologies tests, irrespective of the teaching methodology employed ( p > 0.05). No differences between the teaching methodologies studied ( p > 0.05) were found regarding the amount of correct answers. However, the students who participated in the active (e-learning and problem-based learning) and hybrid teaching methodology reported a positive impact in relation to their diagnostic skills at the questionnaire., Conclusions: All the tested methodologies had a similar performance; however, the traditional methodology showed less acceptance by the students when analyzed subjectively and comparatively. The results of the present study increase comprehension about teaching methodologies for radiographic diagnosis of proximal carious lesions, and there is a potential to build on the knowledge base and provide practical support for students and educators.

In its 30 years of existence, there are still many improvement possibilities in studies performing the life cycle assessment (LCA) of wastewater treatment plants (WWTPs). Hence, this paper aims to start a guideline development for LCA of urban WWTPs based on the information available in the scientific literature on the topic. The authors used the ProKnow-C systematic review methodology for paper selection and 111 studies were analyzed. The most significant points that can be improved are caused by missing essential information (e.g. functional unity and input data). Other important methodological aspects are covered: allocation process, functional unit choice, sensitivity analysis, and important fluxes to be considered. Many opportunities within the LCA of WWTPs were identified, such as optimization of WWTP operational aspects and resource recovery. Furthermore, LCA should be combined with other methodologies such as big data, data envelopment analysis, life cycle cost assessment, and social life cycle assessment. To achieve this potential, it is clear that the scientific and technical community needs to converge on a new protocol to ensure that LCA application becomes more reliable and transparent.

The treatment for locally advanced rectal carcinomas (LARC) is based on neoadjuvant chemoradiotherapy (nCRT) and surgery, which results in pathological complete response (pCR) in up to 30% of patients. Since epigenetic changes may influence response to therapy, we aimed to identify DNA methylation markers predictive of pCR in LARC patients treated with nCRT. We used high-throughput DNA methylation analysis of 32 treatment-naïve LARC biopsies and five normal rectal tissues to explore the predictive value of differentially methylated (DM) CpGs. External validation was carried out with The Cancer Genome Atlas-Rectal Adenocarcinoma (TCGA-READ 99 cases). A classifier based on three-CpGs DM (linked to OBSL1 , GPR1 , and INSIG1 genes) was able to discriminate pCR from incomplete responders with high sensitivity and specificity. The methylation levels of the selected CpGs confirmed the predictive value of our classifier in 77 LARCs evaluated by bisulfite pyrosequencing. Evaluation of external datasets (TCGA-READ, GSE81006, GSE75546, and GSE39958) reproduced our results. As the three CpGs were mapped near to regulatory elements, we performed an integrative analysis in regions associated with predicted cis-regulatory elements. A positive and inverse correlation between DNA methylation and gene expression was found in two CpGs. We propose a novel predictive tool based on three CpGs potentially useful for pretreatment screening of LARC patients and guide the selection of treatment modality.

Women gradually lose bone from the age of ~35 years, but around menopause, the rate of bone loss escalates due to increasing bone resorption and decreasing bone formation levels, rendering these individuals more prone to developing osteoporosis. The increased osteoclast activity has been linked to a reduced estrogen level and other hormonal changes. However, it is unclear whether intrinsic changes in osteoclast precursors around menopause can also explain the increased osteoclast activity. Therefore, we set up a protocol in which CD14 + blood monocytes were isolated from 49 female donors (40-66 years old). Cells were differentiated into osteoclasts, and data on differentiation and resorption activity were collected. Using multiple linear regression analyses combining in vitro and in vivo data, we found the following: (1) age and menopausal status correlate with aggressive osteoclastic bone resorption in vitro; (2) the type I procollagen N-terminal propeptide level in vivo inversely correlates with osteoclast resorption activity in vitro; (3) the protein level of mature cathepsin K in osteoclasts in vitro increases with age and menopause; and (4) the promoter of the gene encoding the dendritic cell-specific transmembrane protein is less methylated with age. We conclude that monocytes are "reprogrammed" in vivo, allowing them to "remember" age, the menopausal status, and the bone formation status in vitro, resulting in more aggressive osteoclasts. Our discovery suggests that this may be mediated through DNA methylation. We suggest that this may have clinical implications and could contribute to understanding individual differences in age- and menopause-induced bone loss., Competing Interests: Competing interestsThe authors declare no competing interests., (© The Author(s) 2020.)

Kartagener syndrome (KS) is a rare congenital disorder related to bronchiectasis, chronic sinusitis, and situs inversus, predisposing patients to recurrent respiratory infections that can evolve to end-stage lung disease; lung transplantation (LTx) is one of the therapeutic options. This study highlights some concerns in this group of patients, mainly related to the difficulty of performing the transplantation in recipients with suppurative disease and situs inversus. We conducted a retrospective analysis of all KS patients who underwent LTx at 2 national reference centers by the same LTx team. During 29 years of analysis, we performed 12 cases of bilateral sequential LTx in KS patients, representing 2.4% of all Ltx that we performed. Special perioperative care is needed, including vascular access sites and lung isolation techniques; operative concerns include the arteriotomy and bronchotomy during the back table preparation of the graft and concern about the length of the arterial and bronchial anastomosis. We found a higher incidence of bronchial complications is this group that had not been previously reported. Bilateral sequential orthotopic LTx is feasible in this group of patients, and more studies are needed to understand possible reasons for the apparent higher incidence of bronchial complications., (Copyright © 2020. Published by Elsevier Inc.)