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Disability is common in patients with mental disorders, and therapeutic approaches should promote functional recovery to achieve the highest quality of life. This study reports changes in WHODAS 2.0 scores at 1 year of follow-up among patients receiving care from a single mental-health nurse in a community setting in Spain and compare these changes by diagnosis (SMI vs. CMI). Baseline and 1-year follow-up scores were compared. For all participants, we collected sociodemographic information, clinical diagnosis, and disability scores according to WHODAS 2.0. Participating patients received the usual, non-manualized nursing intervention. Our sample comprised 133 patients, 75 of whom were assessed after 1 year. All patients had improved WHODAS 2.0 scores (difference of 9.20 ± 2.42; p  < .001) in all domains except those domains related to work ("Life activities") and relationships ("Getting along"). Furthermore, almost half of the patients studied were no longer undergoing treatment with the nurse at 1-year follow-up.

Leiomyosarcoma (LMS) is an aggressive, often poorly differentiated cancer of the smooth muscle (SM) lineage for which the molecular drivers of transformation and progression are poorly understood. In microRNA (miRNA) profiling studies, miR-130b was previously found to be upregulated in LMS vs. normal SM, and down-regulated during the differentiation of mesenchymal stem cells (MSCs) into SM, suggesting a role in LMS tumor progression. In the present study, the effects of miR-130b on human LMS tumorigenesis were investigated. Stable miR-130b overexpression enhanced invasion of LMS cells in vitro, and led to the formation of undifferentiated, pleomorphic tumors in vivo, with increased growth and metastatic potential compared to control LMS cells. TSC1 was identified as a direct miR-130b target in luciferase-3'UTR assays, and shRNA-mediated knockdown of TSC1 replicated miR-130b effects. Loss-of-function and gain-of-function studies showed that miR-130b levels regulate cell morphology and motility. Following miR-130b suppression, LMS cells adopted a rounded morphology, amoeboid mode of cell movement and enhanced invasive capacity that was Rho/ROCK dependent. Conversely, miR-130b-overexpressing LMS cells exhibited Rho-independent invasion, accompanied by down-regulation of Rho-pathway effectors. In mesenchymal stem cells, both miR-130b overexpression and TSC1 silencing independently impaired SM differentiation in vitro. Together, the data reveal miR-130b as a pro-oncogenic miRNA in LMS and support a miR-130b-TSC1 regulatory network that enhances tumor progression via inhibition of SM differentiation., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: Memorial Sloan Kettering Cancer Center and one investigator involved in this study (M.O.) have financial interests in Elucida Oncology. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2023 Danielson 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.)

Abstract: Serrano, C, Felipe, JL, García-Unanue, J, Vicente Gimenez, J, Jiménez-Linares, L, Ibáñez, E, Hernando, E, Gallardo, L, and Sánchez-Sánchez, J. Modeling dynamical positional physical data on field zones occupied by playing positions in elite-level futsal: a comparison between running velocities, acceleration, and deceleration. J Strength Cond Res 37(1): 200-206, 2023-The aim of this study was to analyze the influence of playing positions on the physical demands and the specific court zones occupied during official futsal games. The total number of observations were 188, corresponding to 8 matches from the first division of the Spanish Futsal League during the 2019-2020 season. All dynamic motion representations were recorded using an 18 Hz device with ultra-wideband technology and local positioning system installed on the futsal pitch for each position. Python Data Analysis Library, Matplotlib, and Python programming language were applied as computational tools to generate visual analysis of figures to represent every physical demand analyzed. An analysis of variance test showed differences between playing positions, specifically, for winger position (p < 0.05). Based on eta-square coefficients, all variables showed large effects, except acceleration distance zone 1 (1-2 m·s-2). Accordingly, the visual representation of positional dynamic motion data showed differences in the physical demands and the court zone occupied by the different playing positions. This innovative visual analysis allows fitness coaches to associate the physical demands with court zones occupied by different playing positions, and it aids in the design of training drills associated with futsal competition requirements., (Copyright © 2021 National Strength and Conditioning Association.)

Melanoma is an aggressive skin cancer reliant on early detection for high likelihood of successful treatment. Solar UV exposure transforms melanocytes into highly mutated tumor cells that metastasize to the liver, lungs, and brain. Even upon resection of the primary tumor, almost thirty percent of patients succumb to melanoma within twenty years. Identification of key melanoma genetic drivers led to the development of pharmacological BRAF V600E and MEK inhibitors, significantly improving metastatic patient outcomes over traditional cytotoxic chemotherapy or pioneering IFN-α and IL-2 immune therapies. Checkpoint blockade inhibitors releasing the immunosuppressive effects of CTLA-4 or PD-1 proved to be even more effective and are the standard first-line treatment. Despite these major improvements, durable responses to immunotherapy and targeted therapy have been hindered by intrinsic or acquired resistance. In addition to gained or selected genetic alterations, cellular plasticity conferred by epigenetic reprogramming is emerging as a driver of therapy resistance. Epigenetic regulation of chromatin accessibility drives gene expression and establishes distinct transcriptional cell states. Here we review how aberrant chromatin, transcriptional, and epigenetic regulation contribute to therapy resistance and discuss how targeting these programs sensitizes melanoma cells to immune and targeted therapies.

It is unclear why some melanomas aggressively metastasize while others remain indolent. Available studies employing multi-omic profiling of melanomas are based on large primary or metastatic tumors. We examine the genomic landscape of early-stage melanomas diagnosed prior to the modern era of immunological treatments. Untreated cases with Stage II/III cutaneous melanoma were identified from institutions throughout the United States, Australia and Spain. FFPE tumor sections were profiled for mutation, methylation and microRNAs. Preliminary results from mutation profiling and clinical pathologic correlates show the distribution of four driver mutation sub-types: 31% BRAF; 18% NRAS; 21% NF1; 26% Triple Wild Type. BRAF mutant tumors had younger age at diagnosis, more associated nevi, more tumor infiltrating lymphocytes, and fewer thick tumors although at generally more advanced stage. NF1 mutant tumors were frequent on the head/neck in older patients with severe solar elastosis, thicker tumors but in earlier stages. Triple Wild Type tumors were predominantly male, frequently on the leg, with more perineural invasion. Mutations in TERT, TP53, CDKN2A and ARID2 were observed often, with TP53 mutations occurring particularly frequently in the NF1 sub-type. The InterMEL study will provide the most extensive multi-omic profiling of early-stage melanoma to date. Initial results demonstrate a nuanced understanding of the mutational and clinicopathological landscape of these early-stage tumors., (© 2022 The Authors. Pigment Cell & Melanoma Research published by John Wiley & Sons Ltd.)

Brain metastases are the most common brain malignancy. This review discusses the studies presented at the third annual meeting of the Melanoma Research Foundation in the context of other recent reports on the biology and treatment of melanoma brain metastases (MBM). Although symptomatic MBM patients were historically excluded from immunotherapy trials, efforts from clinicians and patient advocates have resulted in more inclusive and even dedicated clinical trials for MBM patients. The results of checkpoint inhibitor trials were discussed in conversation with current standards of care for MBM patients, including steroids, radiotherapy, and targeted therapy. Advances in the basic scientific understanding of MBM, including the role of astrocytes and metabolic adaptations to the brain microenvironment, are exposing new vulnerabilities which could be exploited for therapeutic purposes. Technical advances including single-cell omics and multiplex imaging are expanding our understanding of the MBM ecosystem and its response to therapy. This unprecedented level of spatial and temporal resolution is expected to dramatically advance the field in the coming years and render novel treatment approaches that might improve MBM patient outcomes., (© 2022 The Authors. Pigment Cell & Melanoma Research published by John Wiley & Sons Ltd.)

Melanoma brain metastasis (MBM) frequently occurs in patients with advanced melanoma; yet, our understanding of the underlying salient biology is rudimentary. Here, we performed single-cell/nucleus RNA-seq in 22 treatment-naive MBMs and 10 extracranial melanoma metastases (ECMs) and matched spatial single-cell transcriptomics and T cell receptor (TCR)-seq. Cancer cells from MBM were more chromosomally unstable, adopted a neuronal-like cell state, and enriched for spatially variably expressed metabolic pathways. Key observations were validated in independent patient cohorts, patient-derived MBM/ECM xenograft models, RNA/ATAC-seq, proteomics, and multiplexed imaging. Integrated spatial analyses revealed distinct geography of putative cancer immune evasion and evidence for more abundant intra-tumoral B to plasma cell differentiation in lymphoid aggregates in MBM. MBM harbored larger fractions of monocyte-derived macrophages and dysfunctional TOX + CD8 + T cells with distinct expression of immune checkpoints. This work provides comprehensive insights into MBM biology and serves as a foundational resource for further discovery and therapeutic exploration., Competing Interests: Declaration of interests B.I. has received honoraria from consulting with Merck, Janssen Pharmaceuticals, Astra Zeneca, and Volastra Therapeutics. M.A.D. has been a consultant to Roche/Genentech, Array, Pfizer (New York, NY, United States of America), Novartis, BMS, GSK, Sanofi-Aventis (Bridgewater, NJ, United States of America), Vaccinex, Apexigen, EISAI, and ABM Therapeutics and he has been the PI of research grants to MD Anderson by Roche/Genentech (South San Francisco, CA, United States of America), GSK, Sanofi-Aventis, Merck, Myriad, and Oncothyreon. A.R. has received honoraria from consulting with CStone, Merck, and Vedanta, is or has been a member of the scientific advisory board and holds stock in Advaxis, Appia, Apricity, Arcus, Compugen, CytomX, Highlight, ImaginAb, ImmPact, ImmuneSensor, Inspirna, Isoplexis, Kite-Gilead, Lutris, MapKure, Merus, PACT, Pluto, RAPT, Synthekine, and Tango, has received research funding from Agilent (Santa Clara, CA, United States of America) and from Bristol-Myers Squibb through Stand Up to Cancer (SU2C), and patent royalties from Arsenal Bio. T.E. has acted as a consultant for Almiral Hermal, Bristol-Myers Squibb, MSD, Novartis, Pierre Fabre, and Sanofi. E.Z.M. is a consultant for Curio Bioscience. The other authors do not declare competing interests., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Introduction: Mental disorders are among the leading causes of disability worldwide. Despite the fact that severe mental disorders (SMD) are associated with high disability, the impact of common mental disorders (CMD) is not negligible. In this work, we compare the disability measured with the WHODAS 2.0 scale of both diagnostic groups at the Mental Health Nurse facility., Material and Methods: Sociodemographic data, clinical diagnosis and disability scores were collected, using the WHODAS 2.0 scale, of the patients attended by the Mental Health specialist nurse at the Infanta Elena de Valdemoro Hospital (Madrid) and disability was compared in patients with SMD and CMD, using the Student t test., Results: Our study sample consisted of 133 patients. Patients with CMD showed greater disability compared to patients with SMD. It was observed that the disability associated with CMD is higher, compared to SMD, this difference being significant for the domain of work (p < 0.001) and participation in society (p = 0.041)., Conclusions: In this study we showed that the level of disability associated with CMD was higher in certain areas compared to SMD, this difference was of special relevance for the «Work» and «Participation» domains. This may serve to adapt the interventions aimed at these people and improve their quality of life., (Copyright © 2022. Published by Elsevier España, S.L.U.)

The molecular basis of amyloid toxicity in Alzheimer's disease (AD) remains controversial. Amyloid β (Aβ) oligomers promote Ca 2+ influx, mitochondrial Ca 2+ overload and apoptosis in hippocampal neurons in vivo and in vitro, but the primary Ca 2+ entry pathways are unclear. We studied Ca 2+ entry pathways induced by Aβ oligomers in rat hippocampal and cerebellar neurons. Aβ oligomers induce Ca 2+ entry in neurons. Ca 2+ responses to Aβ oligomers are large after synaptic networking and prevented by blockers of synaptic transmission. In contrast, in neurons devoid of synaptic connections, Ca 2+ responses to Aβ oligomers are small and prevented only by blockers of amyloid channels (NA7) and NMDA receptors (MK801). A combination of NA7 and MK801 nearly abolished Ca 2+ responses. Non-neuronal cells bearing NMDA receptors showed Ca 2+ responses to oligomers, whereas cells without NMDA receptors did not exhibit Ca 2+ responses. The expression of subunits of the NMDA receptor NR1/ NR2A and NR1/NR2B in HEK293 cells lacking endogenous NMDA receptors restored Ca 2+ responses to NMDA but not to Aβ oligomers. We conclude that Aβ oligomers promote Ca 2+ entry via amyloid channels and NMDA receptors. This may recruit distant neurons intertwisted by synaptic connections, spreading excitation and recruiting further NMDA receptors and voltage-gated Ca 2+ channels, leading to excitotoxicity and neuron degeneration in AD.

Brain metastasis is a significant cause of morbidity and mortality in multiple cancer types and represents an unmet clinical need. The mechanisms that mediate metastatic cancer growth in the brain parenchyma are largely unknown. Melanoma, which has the highest rate of brain metastasis among common cancer types, is an ideal model to study how cancer cells adapt to the brain parenchyma. Our unbiased proteomics analysis of melanoma short-term cultures revealed that proteins implicated in neurodegenerative pathologies are differentially expressed in melanoma cells explanted from brain metastases compared with those derived from extracranial metastases. We showed that melanoma cells require amyloid beta (Aβ) for growth and survival in the brain parenchyma. Melanoma-secreted Aβ activates surrounding astrocytes to a prometastatic, anti-inflammatory phenotype and prevents phagocytosis of melanoma by microglia. Finally, we demonstrate that pharmacologic inhibition of Aβ decreases brain metastatic burden., Significance: Our results reveal a novel mechanistic connection between brain metastasis and Alzheimer's disease, two previously unrelated pathologies; establish Aβ as a promising therapeutic target for brain metastasis; and demonstrate suppression of neuroinflammation as a critical feature of metastatic adaptation to the brain parenchyma. This article is highlighted in the In This Issue feature, p. 1171., (©2022 American Association for Cancer Research.)

Melanoma is a highly prevalent cancer with an increasing incidence worldwide and high metastatic potential. Brain metastasis is a major complication of the disease, as more than 50% of metastatic melanoma patients eventually develop intracranial disease. MicroRNAs (miRNAs) have been found to play an important role in the tumorigenicity of different cancers and have potential as markers of disease outcome. Identification of relevant miRNAs has generally stemmed from miRNA profiling studies of cells or tissues, but these approaches may have missed miRNAs with relevant functions that are expressed in subfractions of cancer cells. We performed an unbiased in vivo screen to identify miRNAs with potential functions as metastasis suppressors using a lentiviral library of miRNA decoys. Notably, we found that a significant fraction of melanomas that metastasized to the brain carried a decoy for miR-124a, a miRNA that is highly expressed in the brain/neurons. Additional loss- and gain-of-function in vivo validation studies confirmed miR-124a as a suppressor of melanoma metastasis and particularly of brain metastasis. miR-124a overexpression did not inhibit tumor growth in vivo , underscoring that miR-124a specifically controls processes required for melanoma metastatic growth, such as seeding and growth post-extravasation. Finally, we provide proof of principle of this miRNA as a promising therapeutic agent by showing its ability to impair metastatic growth of melanoma cells seeded in distal organs. Our efforts shed light on miR-124a as an antimetastatic agent, which could be leveraged therapeutically to impair metastatic growth and improve patient survival., 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 © 2022 Moubarak, Koetz-Ploch, Mullokandov, Gaziel, de Pablos-Aragoneses, Argibay, Kleffman, Sokolova, Berwick, Thomas, Osman, Brown and Hernando.)

Metastasis is a complex process, requiring cells to overcome barriers that are only incompletely modeled by in vitro assays. A systematic workflow was established using robust, reproducible in vivo models and standardized methods to identify novel players in melanoma metastasis. This approach allows for data inference at specific experimental stages to precisely characterize a gene's role in metastasis. Models are established by introducing genetically modified melanoma cells via intracardiac, intradermal, or subcutaneous injections into mice, followed by monitoring with serial in vivo imaging. Once preestablished endpoints are reached, primary tumors and/or metastases-bearing organs are harvested and processed for various analyses. Tumor cells can be sorted and subjected to any of several 'omics' platforms, including single-cell RNA sequencing. Organs undergo imaging and immunohistopathological analyses to quantify the overall burden of metastases and map their specific anatomic location. This optimized pipeline, including standardized protocols for engraftment, monitoring, tissue harvesting, processing, and analysis, can be adopted for patient-derived, short-term cultures and established human and murine cell lines of various solid cancer types.

The contribution of epigenetic dysregulation to metastasis remains understudied. Through a meta-analysis of gene expression datasets followed by a mini-screen, we identified Plant Homeodomain Finger protein 8 (PHF8), a histone demethylase of the Jumonji C protein family, as a previously unidentified prometastatic gene in melanoma. Loss- and gain-of-function approaches demonstrate that PHF8 promotes cell invasion without affecting proliferation in vitro and increases dissemination but not subcutaneous tumor growth in vivo, thus supporting its specific contribution to the acquisition of metastatic potential. PHF8 requires its histone demethylase activity to enhance melanoma cell invasion. Transcriptomic and epigenomic analyses revealed that PHF8 orchestrates a molecular program that directly controls the TGFβ signaling pathway and, as a consequence, melanoma invasion and metastasis. Our findings bring a mechanistic understanding of epigenetic regulation of metastatic fitness in cancer, which may pave the way for improved therapeutic interventions.

Background and Objectives : The influence of smoking habits on mortality, VTE recurrence, and major bleeding in patients receiving anticoagulant therapy for venous thromboembolism (VTE) has not been consistently evaluated. Materials and Methods : We used data from the RIETE (Registro Enfermedad TromboEmbólica) registry to compare mortality, VTE recurrence, and major bleeding risk in smoking versus non-smoking patients with acute VTE. Results : 50,881 patients (43,426 non-smoking and 7455 smoking patients) were included. After a median follow-up of 8.8 months, 7110 patients died (fatal PE 292 and fatal bleeding 281), 3243 presented VTE recurrence, and 1579 had major bleeding. At multivariate analysis, smoking behavior was associated with a higher hazard of death, (HR: 1.28; 95% CI: 1.19-1.40). The risk of VTE recurrence was marginally increased in smoking patients compared to non-smoking patients (1.14; 95% CI: 1.02-1.27). Major bleeding did not differ in smoking and non-smoking patients (1.15; 95% CI: 0.96-1.38). The presence of cancer did not appear to influence the association between smoking habits and death (HR: 1.34; 95% CI: 1.22-1.47 in cancer patients and HR: 1.23; 95% CI: 1.04, 1.45 in non-cancer patients, respectively) Conclusions : the risk of death after an acute episode of VTE appeared to be higher in smoking than in non-smoking patients and this risk is higher between patients presenting PE at the onset of symptoms.

Introduction: Mental disorders are among the leading causes of disability worldwide. Despite the fact that severe mental disorders (SMD) are associated with high disability, the impact of common mental disorders (CMD) is not negligible. In this work, we compare the disability measured with the WHODAS 2.0 scale of both diagnostic groups at the Mental Health Nurse facility., Material and Methods: Sociodemographic data, clinical diagnosis and disability scores were collected, using the WHODAS 2.0 scale, of the patients attended by the Mental Health specialist nurse at the Infanta Elena de Valdemoro Hospital (Madrid) and disability was compared in patients with SMD and CMD, using the Student t test., Results: Our study sample consisted of 133 patients. Patients with CMD showed greater disability compared to patients with SMD. It was observed that the disability associated with CMD is higher, compared to SMD, this difference being significant for the domain of work (p<0.001) and participation in society (p=0.041)., Conclusions: In this study we showed that the level of disability associated with CMD was higher in certain areas compared to SMD, this difference was of special relevance for the «Work» and «Participation» domains. This may serve to adapt the interventions aimed at these people and improve their quality of life., (Copyright © 2021. Publicado por Elsevier España, S.L.U.)

Introduction: functional gastrointestinal disorders are prevalent and resource consuming. The use of group-consultations in these diseases is limited and no specific multidisciplinary programs have been developed., Methods: a multidisciplinary approach was used in patients with diverse functional gastrointestinal disorders attending group-consultations (group A). Five two-hour sessions were scheduled over a four-month period. Sessions consisted of a theoretical introduction (Pathophysiology, Low fodmap diet, Over the counter medications, Mediterranean diet, and Laughter therapy workshop) followed by relaxation techniques. This group was compared to a similar group of patients who received written information covering the topics discussed during the group-consultations (group B). Severity of digestive and psychological symptoms, use of drugs and adherence to the diet were the main outcomes measured., Results: the mean age of participants was 43 (± 1.38) years, 78 % were female and 73 % had at least two functional gastrointestinal disorders. Sixty-two patients were included in group A and 17 in group B. The severity of gastrointestinal and psychological symptoms at baseline was similar in both groups. Globally, there was an improvement in all symptoms in both groups. The proportion of participants with severe baseline gastrointestinal symptoms or pathologic anxiety scores that showed improvement was significantly higher in group A (74 % vs 23 %, p = 0.005; 47 % vs 8 %, p = 0.02, respectively). Symptoms were reassessed at six and 12 months after the intervention in participants from group A who attended ≥ 80 % sessions and a sustained response was observed., Conclusions: group-consultations are useful and efficient to alleviate gastrointestinal and psychological symptoms in patients with functional gastrointestinal disorders.

Brain tumors comprise a large series of tumor cancer from benign to highly malignant gliomas and metastases from primary tumors outside the brain. Intracellular Ca 2+ homeostasis is involved in a large series of cell functions including cell proliferation, migration, and cell death. Store-operated Ca 2+ entry (SOCE), the most important Ca 2+ entry pathway in non-excitable cells, is involved in cell proliferation and migration and enhanced in tumor cells from breast cancer, colon cancer and cell lines derived from glioblastoma but there are almost no studies in human primary glioblastoma cells or other brain tumors. We have developed a single procedure to obtain primary cells from a large series (n = 49) of human brain tumors including schwannomas, meningiomas, oligodendrogliomas, astrocytomas, glioblastomas and brain metastases from ovary, breast and lung. Cells were characterized by immunofluorescence and subjected to Ca 2+ imaging to investigate resting intracellular Ca 2+ levels, Ca 2+ responses to physiological agonists as well as voltage-operated Ca 2+ entry and SOCE. We found significant differences in resting intracellular Ca 2+ and Ca 2+ responses to plasma membrane depolarization and ATP among the different tumor cells. Only malignant tumor cells, displayed Ca 2+ responses to ATP. SOCE is significantly increased in malignant gliomas whereas voltage-gated Ca 2+ entry is decreased. In addition, SOCE is significantly larger in high grade gliomas than in low grade gliomas suggesting that SOCE increases with glioma progression. These data may provide new insights on the role of intracellular Ca 2+ and purinergic signalling in brain tumors., (Copyright © 2021. Published by Elsevier B.V.)

Background: Proteinuria is assumed to be less frequent in cats than in dogs and is mainly associated with chronic kidney disease (CKD)., Aim: The current study aimed to evaluate and compare urine protein-to-creatinine (UPC) values retrospectively in cats visited for comprehensive annual health check or for presenting systemic clinical signs related to CKD., Methods: UPC ratio was retrospectively evaluated in 112 owned cats, out of which 51 (45.5%) were apparently healthy cats according to their owners who visited for comprehensive annual health checks and 61 (54.5%) sick cats, presenting systemic clinical signs suggesting CKD, such as weight loss or polyuria/polydipsia, among others., Results: Based on UPC, the present study found that 54.5% of all cats included were borderline proteinuric or proteinuric, having increased UPC (UPC ≥ 0.2), with 35.7% included in the sick group and 18.7% in the health-check group. Increased UPC was also statistically associated with azotemia and isosthenuria (urinary-specific gravity between 1,008 and 1,035) in both sick and health-check groups of cats., Conclusion: Independent of the reason for their medical visit, it could be concluded that borderline proteinuria and proteinuria were statistically mainly related to CKD in cats. Furthermore, the measurement of UPC could be very useful in the detection and management of CKD in apparently healthy cats during a medical visit for annual health check irrespective of the age., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

High spliceosome activity is a dependency for cancer cells, making them more vulnerable to perturbation of the splicing machinery compared to normal cells. To identify splicing factors important for prostate cancer (PCa) fitness, we performed pooled shRNA screens in vitro and in vivo. Our screens identified heterogeneous nuclear ribonucleoprotein M (HNRNPM) as a regulator of PCa cell growth. RNA- and eCLIP-sequencing identified HNRNPM binding to transcripts of key homeostatic genes. HNRNPM binding to its targets prevents aberrant exon inclusion and backsplicing events. In both linear and circular mis-spliced transcripts, HNRNPM preferentially binds to GU-rich elements in long flanking proximal introns. Mimicry of HNRNPM-dependent linear-splicing events using splice-switching-antisense-oligonucleotides was sufficient to inhibit PCa cell growth. This suggests that PCa dependence on HNRNPM is likely a result of mis-splicing of key homeostatic coding and non-coding genes. Our results have further been confirmed in other solid tumors. Taken together, our data reveal a role for HNRNPM in supporting cancer cell fitness. Inhibition of HNRNPM activity is therefore a potential therapeutic strategy in suppressing growth of PCa and other solid tumors., Competing Interests: JH, FD, MS, DL, DI, FG, TT, JZ, HW, LC, OA, TC, AH, SZ, VR, SC, AK, MA, HH, BG, SO, MS, GG, KM, EH, DM, IM, DW, EG No competing interests declared, (© 2021, Ho et al.)

Bacterial urinary tract infection (UTI) is a common clinical concern in dogs. However, incidence of feline UTI is much lower than in dogs although an increasing prevalence has been registered. The main objective of the present study was to describe and characterize the prevalence of urinary tract pathogens in urine samples of dogs and cats with urinary clinical signs throughout different Spanish provinces. Secondary aims were to determine if there were differences in urine sample characterizations based on species (i.e., dog and cat) or season. Dogs were found to have a higher rate of positive urinary cultures than cats (39.3% and 24.7% of the cultures submitted, respectively). The bacterial genera most commonly isolated in dogs were Escherichia spp. (45.3%), Proteus spp. (13.2%), Staphylococcus spp. (11%), and Enterococcus spp. (8.6%). Whereas in the feline population, Escherichia spp. (42.7%), Enterococcus spp. (22.2%), and Staphyloccoccus spp. (15.2%) were the most frequently isolated bacteria. The highest rates of positive urine cultures were registered in Melilla (70%), Zamora (66.7%), Teruel (64.3%), and Guadalajara (60%). Moreover, the proportion of positive urine cultures was not homogeneously distributed across provinces. Finally, some seasonality was found among most isolated bacterias. Enterococcus spp. was significantly more prevalent in summer, whereas Escherichia spp. and Proteus spp. were more commonly isolated in spring and Pseudomonas spp. in autumn., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Five years ago, the Melanoma Research Foundation (MRF) conducted an assessment of the challenges and opportunities facing the melanoma research community and patients with melanoma. Since then, remarkable progress has been made on both the basic and clinical research fronts. However, the incidence, recurrence, and death rates for melanoma remain unacceptably high and significant challenges remain. Hence, the MRF Scientific Advisory Council and Breakthrough Consortium, a group that includes clinicians and scientists, reconvened to facilitate intensive discussions on thematic areas essential to melanoma researchers and patients alike, prevention, detection, diagnosis, metastatic dormancy and progression, response and resistance to targeted and immune-based therapy, and the clinical consequences of COVID-19 for patients with melanoma and providers. These extensive discussions helped to crystalize our understanding of the challenges and opportunities facing the broader melanoma community today. In this report, we discuss the progress made since the last MRF assessment, comment on what remains to be overcome, and offer recommendations for the best path forward., (©2021 American Association for Cancer Research.)

Melanoma is the most lethal skin malignancy, driven by genetic and epigenetic alterations in the complex tumour microenvironment. While large-scale molecular profiling of melanoma has identified molecular signatures associated with melanoma progression, comprehensive systems-level modeling remains elusive. This study builds up predictive gene network models of molecular alterations in primary melanoma by integrating large-scale bulk-based multi-omic and single-cell transcriptomic data. Incorporating clinical, epigenetic, and proteomic data into these networks reveals key subnetworks, cell types, and regulators underlying melanoma progression. Tumors with high immune infiltrates are found to be associated with good prognosis, presumably due to induced CD8+ T-cell cytotoxicity, via MYO1F-mediated M1-polarization of macrophages. Seventeen key drivers of the gene subnetworks associated with poor prognosis, including the transcription factor ZNF180, are tested for their pro-tumorigenic effects in vitro. The anti-tumor effect of silencing ZNF180 is further validated using in vivo xenografts. Experimentally validated targets of ZNF180 are enriched in the ZNF180 centered network and the known pathways such as melanoma cell maintenance and immune cell infiltration. The transcriptional networks and their critical regulators provide insights into the molecular mechanisms of melanomagenesis and pave the way for developing therapeutic strategies for melanoma.

Introduction: In early breast cancer (EBC), a single dose of intraoperative radiotherapy (IORT) might be an option to standard whole breast radiotherapy (WBRT). However, there is no consensus about its use and clinical results., Aim: to analyse the morbidity and oncological outcomes of IORT as monotherapy in EBC., Methods: A single centre observational analytic study was performed. A prospective IORT cohort (2015-17) and a retrospective WBRT cohort (2012-17) were selected following the same criteria: ≥ 45 y.o., invasive ductal carcinoma or variants, radiological tumour size ≤ 3 cm, positive oestrogenic receptors, negative HER2, cN0; exclusion criteria: lymphovascular invasion, multicentricity/multifocality, BRCA mutation and neoadjuvant therapy. Clinical, histological, surgical, oncological characteristics and complications were collected., Results: A total of 425 cases were selected: 217 in IORT cohort and 208 in WBRT cohort. Average age in IORT and WBRT groups was 67±9.5 and 64.8 ± 9.9 y.o. respectively (p = 0.01). ASA 3 risk score patients were 17.7% in IORT and 24 cases in WBRT (p = 0.027). There were no differences in histological results or tumoral stage. Average follow up was 24.4 ± 8 months in IORT and 50.5 ± 18 months in WBRT (p < 0.001). No differences were detected in local recurrence, metastases or mortality. Complications that required reintervention or hospitalization were similar in both groups. A total of 3 and 14 cases developed early severe dermatitis in IORT and WBRT groups respectively (p = 0.01)., Conclusion: IORT as monotherapy in selected patients with EBC stands for an alternative option versus WBRT. It seems especially useful in advanced-age patients with severe comorbidities. IORT associates lesser early severe dermatitis., (Copyright © 2020 AEC. Publicado por Elsevier España, S.L.U. All rights reserved.)

Background: Recent preclinical data suggest that there may be therapeutic synergy between immune checkpoint blockade and inhibition of the coagulation cascade. Here, we investigate whether patients who received immune checkpoint inhibitors (ICI) and were on concomitant anticoagulation (AC) experienced better treatment outcomes than individuals not on AC.Affiliation: Kindly confirm if corresponding authors affiliation is identified correctly.The corresponding author's affiliation is correct., Methods: We studied a cohort of 728 advanced cancer patients who received 948 lines of ICI at NYU (2010-2020). Patients were classified based on whether they did (n = 120) or did not (n = 828) receive therapeutic AC at any point during their treatment with ICI. We investigated the relationship between AC status and multiple clinical endpoints including best overall response (BOR), objective response rate (ORR), disease control rate (DCR), progression free survival (PFS), overall survival (OS), and the incidence of bleeding complications.Affiliations: Journal instruction requires a country for affiliations; however, this is missing in affiliations 1 to 5. Please verify if the provided country is correct and amend if necessary.The country is correct for all affiliations (1 - 5)., Results: Treatment with AC was not associated with significantly different BOR (P = 0.80), ORR (P =0.60), DCR (P =0.77), PFS (P = 0.59), or OS (P =0.64). Patients who received AC were significantly more likely to suffer a major or clinically relevant minor bleed (P = 0.05)., Conclusion: AC does not appear to impact the activity or efficacy of ICI in advanced cancer patients. On the basis of our findings, we caution that there is insufficient evidence to support prospectively evaluating the combination of AC and immunotherapy.

In futsal there are two referees on the playing court and their capacity to respond to physical and physiological demands imposed during the game is essential for the success. The futsal characteristics such as size pitch, referees position and rules of games or type of league could impose specific physical efforts probably. The aim of this study were to analyze the physical demands of eight elite referees (age 40 ± 3.43 years; height 1.80 ± 0.03 m; weight 72.84 ± 4.01 kg) from seven matches of Spanish Futsal Cup 2020. The physical activity of each referee during the match was monitored with a Local Positioning System, which was installed on futsal pitch where the matches were played. The data differences were evaluated as Paired-Samples T -Test procedure. The results revealed a similar total distance between halves (2888.39 vs. 2831.51 m). The zone 3 distance (15.1-18 km⋅h -1 ) showed a significative decrease ( p < 0.05) during the match in comparison to the first and second halves (-24.48 m; CI95%: -9.54 to -39.42; ES: 0.56). The number of high-intensity acceleration (-10.29; CI95%; 3.71-16.86; ES: 0.92) and deceleration (-24.86; CI95%; 11.59-38.12; ES: 0.99) decreased in the second half of the match ( p < 0.05). Therefore, the use of the tracking device to monitoring physical performance provides knowledge of the specific activity profile from futsal referees. This information to can be useful to design more accurate the training programs., 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 © 2021 Serrano, Sánchez-Sánchez, Felipe, Hernando, Gallardo and Garcia-Unanue.)

Purpose: The objectives of the study were to establish a procedure for in vivo film-based dosimetry for intraoperative radiotherapy (IORT), evaluate the typical doses delivered to organs at risk, and verify the dose prescription., Materials and Methods: In vivo dose measurements were studied using XR-RV3 radiochromic films in 30 patients with breast cancer undergoing IORT using the Axxent® device (Xoft Inc.). The stability of the radiochromic films in the energy ranges used was verified by taking measurements at different depths. The stability of the scanner response was tested, and 5 different calibration curves were constructed for different beam qualities. Six pieces of film were placed in each of the 30 patients. All the pieces were correctly sterilized and checked to ensure that the process did not affect the outcome. All calibration and dose measurements were analyzed using the Radiochromic.com software application., Results: The doses were measured for 30 patients. The doses in contact with the applicator (prescription zone) were 19.8 ± 0.9 Gy. In the skin areas, the doses were as follows: 1-2 cm from the applicator, 1.86 ± 0.77 Gy; 2-5 cm, 0.73 ± 0.14 Gy; and greater than 5 cm, 0.28 ± 0.17 Gy. The dose delivered to the pectoral muscle (tungsten shielding disc) was 0.51 ± 0.27 Gy., Conclusions: The study demonstrated the viability of XR-RV3 films for in vivo dose measurement in the dose and energy ranges applied in a complex procedure, such as breast IORT. The doses in organs at risk were far below the tolerances for cases such as those studied., (Copyright © 2021 Associazione Italiana di Fisica Medica. Published by Elsevier Ltd. All rights reserved.)

The aim of this study was to evaluate the neuromuscular response to an agility and repeated sprint ability (RSA) test according to the level of competition in futsal players. A total of 33 players from two elite teams and one amateur team participated in the study. The participants completed an agility t -test, a 30 m-speed test, and a RSA test. A countermovement jump (CMJ) test and a tensiomyography test of the rectus femoris (RF) and biceps femoris (BF) of both legs were carried out before and after the tests. RSA test revealed better sprint times in elite players compared to amateurs in the seven bouts, as well as in the 30 m sprint and in the agility test ( p < 0.05). Before the tests, elite players showed higher sustain time (Ts) in RF (+31.03 ms; ES: 0.76) and BF (+28.73 ms; ES: 0.73), higher half-relaxation time (Tr) in BF (+20.79 ms; ES: 0.94), and lower delay time (Td) in BF (-2 ms; ES: 1.19) compared to amateur players. However, post-test values did not present any significant differences ( p > 0.05). In conclusion, elite players showed greater performance in the RSA test, in the 30 m tests and in the agility test compared to amateur players. The contractile properties were not a key factor in the RSA performance of the futsal players., 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 © 2020 García-Unanue, Felipe, Bishop, Colino, Ubago-Guisado, López-Fernández, Hernando, Gallardo and Sánchez-Sánchez.)

Clinical diagnosis of influenza has low sensitivity in infants and children. Signs and symptoms are non-specific and similar to those of other respiratory viruses. Rapid influenza diagnostic tests (RIDTs) with adequate sensitivity and specificity used at the point of care can be useful for an etiologic diagnosis of influenza in primary care. This should have an impact on better management of these patients. We conducted a study during three consecutive influenza seasons (2016-2017, 2017-2018, and 2018-2019) in pediatric primary care settings collecting data from influenza point-of-care tests (POCTs)-confirmed ≤ 6-year-old patients. During the first two influenza seasons, antibiotic prescriptions and additional visits from influenza POCT-confirmed patients (Group&#95;1) were compared to patients with influenza-like illness (ILI) (Group&#95;2), or fever (2016 ICD-10 code R50) with no other signs of influenza (Group&#95;3). Group&#95;1 had 0.19 (2016-2017) and 0.23 (2017-2018) additional visits compared to 0.48 (2016-2017) and 0.49 (2017-2018) Group&#95;2 p < 0.001 and 1.01 (2016-2017) and 0.80 (2017-2018) Group&#95;3 p < 0.001. Antibiotic prescription was lower in Group&#95;1 (10.2%) vs. Group&#95;3 (17.2%) p < 0.002, difference statistically significant only for the 2017-2018 season. During the third season (2018-2019), RIDTs results were transmitted in real time to the reference laboratory ia the cloud, which strengthens the monitoring of circulating influenza viruses in the community. In our experience, the use of POCTs has a great potential in primary care specially in infants and young children in which the diagnosis maybe missed due to non-specific signs and symptoms., (Copyright © 2020 van Esso, Vilà, Andrés, Iglesias, Ferrer, Losada, Ricos, Sanchez, Morera, Pérez, Ferrandez, Hernando, Rodrigo, Pumarola and Antón.)

We hypothesized that human genes differ by their sensitivity to ultraviolet (UV) exposure. We used somatic mutations detected by genome-wide screens in melanoma and reported in the Catalog Of Somatic Mutations In Cancer. As a measure of UV sensitivity, we used the number of silent mutations generated by C>T transitions in pyrimidine dimers of a given transcript divided by the number of potential sites for this type of mutations in the transcript. We found that human genes varied by UV sensitivity by two orders of magnitude. We noted that the melanoma-associated tumor suppressor gene CDKN2A was among the top five most UV-sensitive genes in the human genome. Melanoma driver genes have a higher UV-sensitivity compared with other genes in the human genome. The difference was more prominent for tumor suppressors compared with oncogene. The results of this study suggest that differential sensitivity of human transcripts to UV light may explain melanoma specificity of some driver genes. Practical significance of the study relates to the fact that differences in UV sensitivity among human genes need to be taken into consideration whereas predicting melanoma-associated genes by the number of somatic mutations detected in a given gene., (© 2020 Wiley Periodicals LLC.)

TSC1 is a tumor suppressor that inhibits cell growth via negative regulation of the mammalian target of rapamycin complex (mTORC1). TSC1 mutations are associated with Tuberous Sclerosis Complex (TSC), characterized by multiple benign tumors of mesenchymal and epithelial origin. TSC1 modulates self-renewal and differentiation in hematopoietic stem cells; however, its effects on mesenchymal stem cells (MSCs) are unknown. We investigated the impact of Tsc1 inactivation in murine bone marrow (BM)-MSCs, using tissue-specific, transgelin ( Tagln )-mediated cre-recombination, targeting both BM-MSCs and smooth muscle cells. Tsc1 mutants were viable, but homozygous inactivation led to a dwarfed appearance with TSC-like pathologies in multiple organs and reduced survival. In young (28 day old) mice, Tsc1 deficiency-induced significant cell expansion of non-hematopoietic BM in vivo, and MSC colony-forming potential in vitro, that was normalized upon treatment with the mTOR inhibitor, everolimus. The hyperproliferative BM-MSC phenotype was lost in aged (1.5 yr) mice, and Tsc1 inactivation was also accompanied by elevated ROS and increased senescence. ShRNA-mediated knockdown of Tsc1 in BM-MSCs replicated the hyperproliferative BM-MSC phenotype and led to impaired adipogenic and myogenic differentiation. Our data show that Tsc1 is a negative regulator of BM-MSC proliferation and support a pivotal role for the Tsc1-mTOR axis in the maintenance of the mesenchymal progenitor pool.

A hallmark of metastasis is the adaptation of tumor cells to new environments. Metabolic constraints imposed by the serine and glycine-limited brain environment restrict metastatic tumor growth. How brain metastases overcome these growth-prohibitive conditions is poorly understood. Here, we demonstrate that 3-phosphoglycerate dehydrogenase (PHGDH), which catalyzes the rate-limiting step of glucose-derived serine synthesis, is a major determinant of brain metastasis in multiple human cancer types and preclinical models. Enhanced serine synthesis proved important for nucleotide production and cell proliferation in highly aggressive brain metastatic cells. In vivo , genetic suppression and pharmacologic inhibition of PHGDH attenuated brain metastasis, but not extracranial tumor growth, and improved overall survival in mice. These results reveal that extracellular amino acid availability determines serine synthesis pathway dependence, and suggest that PHGDH inhibitors may be useful in the treatment of brain metastasis. SIGNIFICANCE: Using proteomics, metabolomics, and multiple brain metastasis models, we demonstrate that the nutrient-limited environment of the brain potentiates brain metastasis susceptibility to serine synthesis inhibition. These findings underscore the importance of studying cancer metabolism in physiologically relevant contexts, and provide a rationale for using PHGDH inhibitors to treat brain metastasis. This article is highlighted in the In This Issue feature, p. 1241 ., (©2020 American Association for Cancer Research.)

The aim of this study was to analyze the influence of the match half and the playing position on physical requirements in the Spanish Professional Futsal League players during official games. The external load from distance, speed, acceleration and deceleration variables were obtained from fourteen elite futsal players during 10 official matches of the 2019-2020 season using a Local Positioning System with ultra-wideband technology installed on the futsal pitch. The results revealed similar results from physical requirements between first and second half ( p > 0.05). Wingers demonstrated greater high-speed running distance (+4.04 m·min -1 ; CI95%: 0.35 to 7.72; ES: 0.87) than pivots ( p > 0.05). There were a high number of accelerations (7.42-9.41 n ·min -1 ) and decelerations (7.37-9.12 n ·min -1 ) per minute in all player positions. The principal finding of the current manuscript did not evidence differences in the physical performance of players between the first and second half. The physical requirements varied among pivots and wingers regarding high-intensity actions. These outcomes add new contributions to the understanding of futsal physical demands.

The ability to effect a change of direction (COD) when performing high-speed actions is essential in team sports like futsal. Nevertheless, the interaction effect of sports surfaces on this ability remains unknown. This research aimed to analyse the plantar pressures and time performance of 15 youth futsal players when performing CODs on two playing surfaces with different mechanical properties. The shock absorption and vertical deformation of one synthetic flooring surface and one wooden flooring surface were assessed. CODs were evaluated using a modified version of the Agility T-Test, while total time of the test, time of CODs, and plantar pressure in CODs were all recorded. The wooden flooring surface displayed higher values of shock absorption (35.70% ± 2.87%) and vertical deformation (2.77 ± 0.38 mm) compared with the synthetic flooring ( p  < 0.01). Faster CODs were performed on the wooden flooring than on the synthetic flooring (-0.05 s, 95% confidence interval [CI]: -0.10 to -0.007, effect size [ES]: 0.07, p  < 0.05), but no differences in total test time were found ( p  > 0.05). Finally, no differences in plantar pressures by playing surface were found. In sum, the differences in the mechanical properties of the two futsal surfaces affected the performance of futsal players in the modified agility test. However, these differences were not great enough to generate different plantar pressures on players, probably due to playerś own adaptations.

As most existing studies in youth academies are focused on top players, the objective of this research is to analyze the physical and physiological demands of various small-sided games (SSGs) on different age categories within a sub-elite soccer academy. We evaluated 63 young players from a Spanish sub-elite academy (under 14 = 21; under 16 = 21; under 18 = 21). Players performed four different small-side games focused on possession game (3-a-side; 4-a-side; 5-a-side; 6-a-side). The global indicators of performance and high-intensity actions were recorded through global positioning systems, whereas the heart rate responses were measured using heart rate monitors. Results: Under 16 ran a greater distance at high-intensity velocity than under 14 in the small side games 3v3 and 6v6. Furthermore, under 16 also ran a greater distance at high-intensity velocity than under 18 in the small side game 3v3 ( p < 0.01). Under 14 showed greater acceleration at the highest intensity (> 2.75 m/s2) than the other age groups, under 16 and U18 ( p < 0.01; ES (effect size) > 1). According to the physiological load, SSG 3v3 presented lower outcomes in Zone 6 (> 95% HRmax) than the small side game 4v4 and the small side game 5v5, in both under 14 and under 16. The workload of SSGs varies depending on the number of players, but also depending on the players' ages. Therefore, when designing the SSGs it is important to consider both the players' ages and the workload that want to be achieved.

This study aimed to investigate morphological, functional, and neuromuscular asymmetries on futsal players' lower limbs at different competitive levels. Sixteen male elite futsal players from the Spanish National Futsal League and thirteen male sub-elite futsal players from the third division participated in this study. Morphological asymmetry was assessed through bioelectrical impedance (fat-mass (g and %) and lean-mass (g)). Functional asymmetry was assessed by means of a 20-second static unipedal balance test. Finally, neuromuscular asymmetry was assessed using tensiomiography tests on both the rectus femoris (RF) and biceps femoris (BF) of each participant. The three tests conducted did not reveal significant bilateral asymmetries in elite players. On the other hand, sub-elite players showed significant bilateral asymmetry in fat-mass percentage between dominant and non-dominant limbs (+6%; CI95%: 1 to 11; ES: 0.88; p = 0.019). They also showed higher bilateral asymmetry in the delay time of the RF (+13%; CI95%: 7 to 21; ES: 1.3; p < 0.05). However, the static unipedal balance test ( p > 0.05) did not evidence asymmetries regardless of the level of the participants. Elite futsal players do not develop bilateral asymmetries in lower limbs in the studied parameters. On the contrary, sub-elite players are likely to develop morphological and neuromuscular asymmetries between their dominant leg and non-dominant leg., Competing Interests: The authors declare no conflict of interest.

Next-generation sequencing has enabled genetic and genomic characterization of melanoma to an unprecedent depth. However, the high mutational background plus the limited depth of coverage of whole-genome sequencing performed on cutaneous melanoma samples make the identification of novel driver mutations difficult. We sought to explore the somatic mutation portfolio in exonic and gene regulatory regions in human melanoma samples, for which we performed targeted sequencing of tumors and matched germline DNA samples from 89 melanoma patients, identifying known and novel recurrent mutations. Two recurrent mutations found in the RPS27 promoter associated with decreased RPS27 mRNA levels in vitro. Data mining and IHC analyses revealed a bimodal pattern of RPS27 expression in melanoma, with RPS27-low patients displaying worse prognosis. In vitro characterization of RPS27-high and RPS27-low melanoma cell lines, as well as loss-of-function experiments, demonstrated that high RPS27 status provides increased proliferative and invasive capacities, while low RPS27 confers survival advantage in low attachment and resistance to therapy. Additionally, we demonstrate that 10 other cancer types harbor bimodal RPS27 expression, and in those, similarly to melanoma, RPS27-low expression associates with worse clinical outcomes. RPS27 promoter mutation could thus represent a mechanism of gene expression modulation in melanoma patients, which may have prognostic and predictive implications., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

By interacting with proteins and nucleic acids, the vast family of mammalian circRNAs is proposed to influence many biological processes. Here, RNA sequencing analysis of circRNAs differentially expressed during myogenesis revealed that circSamd4 expression increased robustly in mouse C2C12 myoblasts differentiating into myotubes. Moreover, silencing circSamd4, which is conserved between human and mouse, delayed myogenesis and lowered the expression of myogenic markers in cultured myoblasts from both species. Affinity pulldown followed by mass spectrometry revealed that circSamd4 associated with PURA and PURB, two repressors of myogenesis that inhibit transcription of the myosin heavy chain (MHC) protein family. Supporting the hypothesis that circSamd4 might complex with PUR proteins and thereby prevent their interaction with DNA, silencing circSamd4 enhanced the association of PUR proteins with the Mhc promoter, while overexpressing circSamd4 interfered with the binding of PUR proteins to the Mhc promoter. These effects were abrogated when using a mutant circSamd4 lacking the PUR binding site. Our results indicate that the association of PUR proteins with circSamd4 enhances myogenesis by contributing to the derepression of MHC transcription., (© Published by Oxford University Press on behalf of Nucleic Acids Research 2020.)

Aging is often associated with a cognitive decline and a susceptibility to neuronal damage. It is also the most important risk factor for neurodegenerative disorders, particularly Alzheimer's disease (AD). AD is related to an excess of neurotoxic oligomers of amyloid β peptide (Aβo); however, the molecular mechanisms are still highly controversial. Intracellular Ca 2+ homeostasis plays an important role in the control of neuronal activity, including neurotransmitter release, synaptic plasticity, and memory storage, as well as neuron cell death. Recent evidence indicates that long-term cultures of rat hippocampal neurons, resembling aged neurons, undergo cell death after treatment with Aβo, whereas short-term cultures, resembling young neurons, do not. These in vitro changes are associated with the remodeling of intracellular Ca 2+ homeostasis with aging, thus providing a simplistic model for investigating Ca 2+ remodeling in aging. In vitro aged neurons show increased resting cytosolic Ca 2+ concentration, enhanced Ca 2+ store content, and Ca 2+ release from the endoplasmic reticulum (ER). Ca 2+ transfer from the endoplasmic reticulum (ER) to mitochondria is also enhanced. Aged neurons also show decreased store-operated Ca 2+ entry (SOCE), a Ca 2+ entry pathway related to memory storage. At the molecular level, in vitro remodeling is associated with changes in the expression of Ca 2+ channels resembling in vivo aging, including changes in N-methyl-D-aspartate NMDA receptor and inositol 1,4,5-trisphosphate (IP 3 ) receptor isoforms, increased expression of the mitochondrial calcium uniporter (MCU), and decreased expression of Orai1/Stim1, the molecular players involved in SOCE. Additionally, Aβo treatment exacerbates most of the changes observed in aged neurons and enhances susceptibility to cell death. Conversely, the solely effect of Aβo in young neurons is to increase ER-mitochondria colocalization and enhance Ca 2+ transfer from ER to mitochondria without inducing neuronal damage. We propose that cultured rat hippocampal neurons may be a useful model to investigate Ca 2+ remodeling in aging and in age-related neurodegenerative disorders.

Metastasis is the primary cause of death of cancer patients. Dissecting mechanisms governing metastatic spread may uncover important tumor biology and/or yield promising therapeutic insights. Here, we investigated the role of circular RNAs (circRNA) in metastasis, using melanoma as a model aggressive tumor. We identified silencing of cerebellar degeneration-related 1 antisense (CDR1as), a regulator of miR-7, as a hallmark of melanoma progression. CDR1as depletion results from epigenetic silencing of LINC00632, its originating long non-coding RNA (lncRNA) and promotes invasion in vitro and metastasis in vivo through a miR-7-independent, IGF2BP3-mediated mechanism. Moreover, CDR1as levels reflect cellular states associated with distinct therapeutic responses. Our study reveals functional, prognostic, and predictive roles for CDR1as and expose circRNAs as key players in metastasis., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Store operated Ca 2+ entry (SOCE) is the most important Ca 2+ entry pathway in non-excitable cells. However, SOCE can also play a pivotal role in excitable cells such as anterior pituitary (AP) cells. The AP gland contains five different cell types that release six major AP hormones controlling most of the entire endocrine system. AP hormone release is modulated by Ca 2+ signals induced by different hypothalamic releasing hormones (HRHs) acting on specific receptors in AP cells. TRH and LHRH both induce Ca 2+ release and Ca 2+ entry in responsive cells while GHRH and CRH only induce Ca 2+ entry. SOCE has been shown to contribute to Ca 2+ responses induced by TRH and LHRH but no molecular evidence has been provided. Accordingly, we used AP cells isolated from mice devoid of Orai1 channels (noted as Orai1-/- or Orai1 KO mice) and mice lacking expression of all seven canonical TRP channels (TRPC) from TRPC1 to TRPC7 (noted as heptaTRPC KO mice) to investigate contribution of these putative channel proteins to SOCE and intracellular Ca 2+ responses induced by HRHs. We found that thapsigargin-evoked SOCE is lost in AP cells from Orai1-/- mice but unaffected in cells from heptaTRPC KO mice. Conversely, while spontaneous intracellular Ca 2+ -oscillations related to electrical activity were not affected in the Orai1-/- mice, these responses were significantly reduced in heptaTRPC KO mice. We also found that Ca 2+ entry induced by TRH and LHRH is decreased in AP cells isolated from Orai1-/-. In addition, Ca 2+ responses to several HRHs, particularly TRH and GHRH, are decreased in the heptaTRPC KO mice. These results indicate that expression of Orai1, and not TRPC channel proteins, is necessary for thapsigargin-evoked SOCE and is required to support Ca 2+ entry induced by TRH and LHRH in mouse AP cells. In contrast, TRPC channel proteins appear to contribute to spontaneous Ca 2+ -oscillations and Ca 2+ responses induced by TRH and GHRH. We conclude that expression of Orai1 and TRPC channels proteins may play differential and significant roles in AP physiology and endocrine control., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The aim of this study was to analyse the influence of age category on the performance and muscle response after a Repeated Sprint Ability (RSA) test in elite youth soccer players. 62 soccer players from three different age categories (Under 14 [ n = 21], Under 16 [ n = 20], and Under 18 [ n = 21]) were selected to participate in this study. Players completed an RSA test (7 × 30 m) with a 20-s recovery between sprints. The muscular response to an electrical stimulus before and after the test of both the biceps femoris (BF) and the rectus femoris (RF) were evaluated using tensiomyography. A two-way ANOVA was used to analyse the differences in RSA parameters in each of the four distance-intervals (0-5; 5-25; 25-30; 0-30 m) between sprint and age category. The U14 age category (5.30 ± 0.30 s) showed higher mean sprint times than U16 (4.62 ± 0.20 s) and U18 (4.46 ± 0.17 s) throughout the entire test ( p < 0.01). U16 players revealed a worse best sprints time (RSA BEST ) than U18 players (+0.12 s, CI95%: to 0.01 to 0.24; ES: 1.09, p = 0.03). The muscular contractile properties were similar in the three age categories analyzed ( p > 0.05), although the delay time (Td) of the muscle was significantly lower after the RSA test in U16 players (-1.53 ms, CI95%: -2.607 to -0.452; ES: 0.38) and U18 players (-1.11 ms, CI95%: -2.10 to -0.12; ES: 0.22). In conclusion, this study revealed an increase in physical performance and muscle response variability after a repeated sprint ability test in the U16's and over. The fatigue induced by the RSA test did not show differences depending on the age of the players, although muscle mechanical properties were altered after the RSA test in U16 and U18 soccer players. Physical performance and muscle response can be complementary variables in managing fatigue according to the age category in soccer players.

Alzheimer's disease (AD) is the most common neurodegenerative disorder and strongly associated to aging. AD has been related to excess of neurotoxic oligomers of amyloid β peptide (Aβo), loss of intracellular Ca 2+ homeostasis and mitochondrial damage. However, the intimate mechanisms underlying the pathology remain obscure. We have reported recently that long-term cultures of rat hippocampal neurons resembling aging neurons are prone to damage induced by Aβ oligomers (Aβo) while short-term cultured cells resembling young neurons are not. In addition, we have also shown that aging neurons display critical changes in intracellular Ca 2+ homeostasis including increased Ca 2+ store content and Ca 2+ transfer from the endoplasmic reticulum (ER) to mitochondria. Aging also promotes the partial loss of store-operated Ca 2+ entry (SOCE), a Ca 2+ entry pathway involved in memory storage. Here, we have addressed whether Aβo treatment influences differentially intracellular Ca 2+ homeostasis in young and aged neurons. We found that Aβo exacerbate the remodeling of intracellular Ca 2+ induced by aging. Specifically, Aβo exacerbate the loss of SOCE observed in aged neurons. Aβo also exacerbate the increased resting cytosolic Ca 2+ concentration, Ca 2+ store content and Ca 2+ release as well as increased expression of the mitochondrial Ca 2+ uniporter (MCU) observed in aging neurons. In contrast, Aβo elicit none of these effects in young neurons. Surprisingly, we found that Aβo increased the Ca 2+ transfer from ER to mitochondria in young neurons without having detrimental effects. Consistently, Aβo increased also colocalization of ER and mitochondria in both young and aged neurons. However, in aged neurons, Aβo suppressed Ca 2+ transfer from ER to mitochondria, decreased mitochondrial potential, enhanced reactive oxygen species (ROS) generation and promoted apoptosis. These results suggest that modulation of ER-mitochondria coupling in hippocampal neurons may be a novel physiological role of Aβo. However, excess of Aβo in the face of the remodeling of intracellular Ca 2+ homeostasis associated to aging may lead to loss of ER-mitochondrial coupling and AD.

Background: Two primary histologic subtypes, superficial spreading melanoma (SSM) and nodular melanoma (NM), comprise the majority of all cutaneous melanomas. NM is associated with worse outcomes, which have been attributed to increased thickness at presentation, and it is widely expected that NM and SSM would exhibit similar behavior once metastasized. Herein, we tested the hypothesis that primary histologic subtype is an independent predictor of survival and may impact response to treatment in the metastatic setting., Methods: We examined the most recent Surveillance, Epidemiology, and End Results (SEER) cohort (n = 118 508) and the New York University (NYU) cohort (n = 1621) with available protocol-driven follow-up. Outcomes specified by primary histology were studied in both the primary and metastatic settings with respect to BRAF-targeted therapy and immunotherapy. We characterized known driver mutations and examined a 140-gene panel in a subset of NM and SSM cases using next-generation sequencing. All statistical tests were two-sided., Results: NM was an independent risk factor for death in both the SEER (hazard ratio [HR] = 1.55, 95% confidence interval [CI] = 1.41 to 1.70, P < .001) and NYU (HR = 1.47, 95% CI = 1.05, 2.07, P = .03) cohorts, controlling for thickness, ulceration, stage, and other variables. In the metastatic setting, NM remained an independent risk factor for death upon treatment with BRAF-targeted therapy (HR = 3.33, 95% CI = 1.06 to 10.47, P = .04) but showed no statistically significant difference with immune checkpoint inhibition. NM was associated with a higher rate of NRAS mutation (P < .001), and high-throughput sequencing revealed NM-specific genomic alterations in NOTCH4, ANK3, and ZNF560, which were independently validated., Conclusions: Our data reveal distinct clinical and biological differences between NM and SSM that support revisiting the prognostic and predictive impact of primary histology subtype in the management of cutaneous melanoma., (© The Author(s) 2018. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

MicroRNAs are critical post-transcriptional regulators of a majority of genes, of which the FOXO family of transcription factors is no exception. Here, we describe generalizable methods, including 3' UTR reporter assays and western blotting after microRNA manipulation, to test if a candidate miRNA (miR-182) directly targets a candidate (FOXO3) gene product. We also provide guidance on candidate miRNA selection and unbiased miRNA-target identification methods.

Melanoma is the most aggressive type of skin cancer in the United States with an increasing incidence. Melanoma lesions often exhibit high immunogenicity, with infiltrating immune cells playing important roles in regression of tumors occurring spontaneously or caused by therapeutic treatment. Computational and experimental methods have been used to estimate the abundance of immune cells in tumors, but their applications are limited by the requirement of large gene sets or multiple antibodies. Although the prognostic role of immune cells has been appreciated, a systematic investigation of their association with clinical factors, genomic features, prognosis and treatment response in melanoma is still lacking. This study, identifies a 25-gene signature based on RNA-seq data from The Cancer Genome Atlas (TCGA)-Skin Cutaneous Melanoma (TCGA-SKCM) dataset. This signature was used to calculate sample-specific Leukocyte Infiltration Scores (LIS) in six independent melanoma microarray datasets and scores were found to vary substantially between different melanoma lesion sites and molecular subtypes. For metastatic melanoma, LIS was prognostic in all datasets with high LIS being associated with good survival. The current approach provided additional prognostic information over established clinical factors, including age, tumor stage, and gender. In addition, LIS was predictive of patient survival in stage III melanoma, and treatment efficacy of tumor-specific antigen vaccine. IMPLICATIONS: This study identifies a 25-gene signature that effectively estimates the level of immune cell infiltration in melanoma, which provides a robust biomarker for predicting patient prognosis., (©2018 American Association for Cancer Research.)

The aim of this study was to evaluate the effect of a repeated sprint ability (RSA) test on the contractile properties of the muscles in elite futsal players. A total of 20 elite players completed the RSA test (7 × 30 m), and the contractile response from the rectus femoris (RF) and biceps femoris (BF) of both legs were analysed pre and post through tensiomyography. There was a significant increment in 30-m times from the third sprint onwards (p < 0.05). The percent decrement in sprint ability (RSA DEC ) with respect to the first sprint was significantly higher in the last sprint. The players did not show evidence of lateral asymmetry in any of the muscle groups analysed after the RSA test (p > 0.05). Following the RSA test there was a significant reduction in the delay time (Td) in RF, a significant decrement in half-relaxation time (Tr) in the RF, and a significant reduction in sustain time (Ts) in the RF and BF of both legs. The maximum radial displacement of the muscle belly (Dm) increased (1.6 mm; effect size = 0.75; p < 0.05) in the RF after the RSA test, indicating reduced muscle stiffness and the ability to generate strength rapidly. The decrement in performance during the RSA test was significantly correlated with changes in contraction time (Tc) in RF and BF, Td in BF, and Dm in RF (p < 0.05). The RSA test generated alterations in the contractile properties of the RF and BF in elite players. However, futsal players did not present asymmetries in any muscular parameters. The baseline contractile muscle parameters could be an important factor related to performance of players during repeated high-intensity actions.

Background: Latest studies suggest similar performance of soccer players either on artificial turf (AT) or natural grass (NG). However, it is not clear if their muscular and physiological responses are also similar on both surfaces. This research aims to assess the influence of game surface on physiological patterns and neuromuscular responses of soccer players during a soccer simulation protocol (SSP) that incorporates repeated sprints and nonlinear actions at maximum speed., Methods: Sixteen amateur soccer players completed three bouts of the SSP on both AT and NG. The mechanical behaviour of both surfaces was recorded and the order was randomly established for each player. The physiological responses were measured during the SSP. A contra-movement jump and a tensiomyography analysis of the rectus femoris (RF) and biceps femoris (BF) were assessed right before and right after the SSP., Results: Both surfaces presented different mechanical properties. No differences among either surfaces or bouts were found for heart rate (HR) peak and HR mean (P>0.05). While the half-relaxation time of the RF on NG decreased after the SSP (right-leg: -44.430 ms; P=0.049; left-leg: -52.131 ms; P=0.008), the sustain time of the BF decreased after the SSP on AT (right-leg: +64.868 ms; P=0.007; left-leg: +87.564 ms; P<0.001). No differences between surfaces were found for the contra-movement jump., Conclusions: The mechanical behaviour of both surfaces does not differ enough to cause different physiological and neuromuscular responses. Playing on AT should cause similar neuromuscular responses to NG.