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Beside inherited muscle diseases many catabolic conditions such as insulin resistance, malnutrition, cancer growth, aging, infections, chronic inflammatory status, inactivity, obesity are characterized by loss of muscle mass, strength and function. The decrease of muscle quality and quantity increases morbidity, mortality and has a major impact on the quality of life. One of the pathogenetic mechanisms of muscle wasting is the dysregulation of the main protein and organelles quality control system of the cell: the autophagy-lysosome. This review will focus on the role of the autophagy-lysosome system in the different conditions of muscle loss. We will also dissect the signalling pathways that are involved in excessive or defective autophagy regulation. Finally, the state of the art of autophagy modulators that have been used in preclinical or clinical studies to ameliorate muscle mass will be also described.

Objective Italy is facing the COVID‐19 outbreak with an abrupt reorganization of its national health‐system, in order to augment care provision to symptomatic patients. The sudden shift of personnel and resources towards COVID‐19 care has led to the reduction of surgery, with possible severe drawbacks. The aim of the study is to describe the trend in surgical volume in urology, in Italy. Materials and Methods Thirty‐three urological units with physicians affiliated to the AGILE consortium were involved in a survey. Urologists were asked to report the amount of surgical elective procedures week‐by‐week, from the beginning of the emergency to the following month. Results The 33 hospitals involved in the study account, globally, for 22,945 beds and are distributed in 13/20 Italian regions. Before the outbreak, the involved urology units performed an overall amount of 1,213 procedures per week, half of which were oncological. One month later, the amount of surgery declined by 78%. Lombardy, the first region with positive‐cases, experienced a 94% reduction. The decrease in oncological and non‐oncological surgical activity was 35,9% and 89%, respectively. The trend of the decline showed a delay of roughly 2 weeks for the other regions. Conclusion Italy, the country with the highest fatality rate from COVID‐19, is experiencing a sudden decline in surgical activity. It is inversely related to the increase in COVID‐19 care, with potential harm particularly in the oncological field. The Italian experience can be helpful for future surgical pre‐planning in other countries not so hardly hit by the disease yet.

Most patients with advanced solid cancers exhibit features of cachexia, a debilitating syndrome characterized by progressive loss of skeletal muscle mass and strength. Because the underlying mechanisms of this multifactorial syndrome are incompletely defined, effective therapeutics have yet to be developed. Here, we show that diminished bone morphogenetic protein (BMP) signaling is observed early in the onset of skeletal muscle wasting associated with cancer cachexia in mouse models and in patients with cancer. Cancer-mediated factors including Activin A and IL-6 trigger the expression of the BMP inhibitor Noggin in muscle, which blocks the actions of BMPs on muscle fibers and motor nerves, subsequently causing disruption of the neuromuscular junction (NMJ), denervation, and muscle wasting. Increasing BMP signaling in the muscles of tumor-bearing mice by gene delivery or pharmacological means can prevent muscle wasting and preserve measures of NMJ function. The data identify perturbed BMP signaling and denervation of muscle fibers as important pathogenic mechanisms of muscle wasting associated with tumor growth. Collectively, these findings present interventions that promote BMP-mediated signaling as an attractive strategy to counteract the loss of functional musculature in patients with cancer.

The dynamic coordination of processes controlling the quality of the mitochondrial network is crucial to maintain the function of mitochondria in skeletal muscle. Changes of mitochondrial proteolytic system, dynamics (fusion/fission), and mitophagy induce pathways that affect muscle mass and performance. When muscle mass is lost, the risk of disease onset and premature death is dramatically increased. For instance, poor quality of muscles correlates with the onset progression of several age-related disorders such as diabetes, obesity, cancer, and aging sarcopenia. To date, there are no drug therapies to reverse muscle loss, and exercise remains the best approach to improve mitochondrial health and to slow atrophy in several diseases. This review will describe the principal mechanisms that control mitochondrial quality and the pathways that link mitochondrial dysfunction to muscle mass regulation.

To retrospectively review our 20 year experience of multidisciplinary management of non-metastatic ductal prostate cancer (dPC), a rare but aggressive histological subtype of prostate cancer whose optimal therapeutic approach is still controversial. Histologically confirmed dPC patients undergoing primary, curative treatment [radical prostatectomy (RP), external beam radiotherapy (EBRT), and androgen deprivation therapy (ADT)] were included, and percentage of ductal and acinar pattern within prostate samples were derived. Survival outcomes were assessed using the subdistribution hazard ratio (SHR) and Fine-and-Gray model. From January 1997 to December 2016, 81 non-metastatic dPC fitted selection criteria. Compared to surgery alone, SHR for progression-free survival and cancer-specific mortality were 2.8 (95% CI 0.6–13.3) and 1.3 (95% CI 0.1–16.2) for exclusive EBRT, 2.7 (95% CI 0.6–13.0) and 6.5 (95% CI 0.6–69.8) for adjuvant EBRT, 4.9 (95% CI 0.7–35.5) and 5.8 (95% CI 0.5–65.6) for salvage EBRT post-prostatectomy recurrence, and 3.2 (95% CI 0.7–14.0) and 3.9 (95% CI 0.3–44.1) for primary ADT (P = 0.558; P = 0.181), respectively. Comparing multimodal treatment and monotherapy confirmed the above trends. Local recurrence more typically occurred in pure dPC patients, mixed histology more frequently produced metastatic spread (29.6% relapse in total, P = 0.026). Albeit some limitations affected the study, our findings support the role of local treatment to achieve better disease control and improve quality of life. Different behavior, with typical local growth in pure dPC, higher distant metastatization in the mixed form, might influence treatment response. Given its poor prognosis, we recommend multidisciplinary management of dPC.

Paola Zuccolotto, Marica Manisera and Marco Sandri give a play-by-play of basketball data analyses to assist students, coaches, technical experts and fans in scrutinising the sport, its teams and players

Purpose To explore the role of vacuum assisted closure (VAC) therapy versus conventional dressings in the Fournier’s gangrene wound therapy. Patients and Methods This is a retrospective multi-institutional cohort study. Data of 92 patients from nine centers between 2007 and 2018 were retrospectively analyzed. After surgery, patient having a local or a disseminated FG were managed with VAC therapy or with conventional dressings. The 10-weeks wound closure cumulative rate and OS were analyzed. Results Of the 92 patients, 62 (67.4%) showed local and 30 (32.6%) a disseminated FG. After surgery, 19 patients (20.7%) with local and 14 (15.2%) with disseminated FG underwent to VAC therapy; 43 (46.7%) with local and 16 (17.4%) with disseminated FG were treated using conventional dressings. The multivariable logistic regression analysis demonstrated that the VAC in patients with disseminated FG led to a higher cumulative rate of wound closure than patients treated with no-VAC (OR = 6.5; 95% CI 1.1–37.4, p = 0.036). The Kaplan–Meier survival curves for the OS showed a significant difference between no-VAC patients with local and disseminated FG (OS rate at 90 days 0.90, 95% CI 0.71–0.97 vs 0.55, 95% CI 0.24–0.78, respectively; p = 0.039). Cox regression confirmed that no-VAC patients with disseminated FG showed the lowest OS (hazard ratio adjusted for sex and age HR = 3.4, 95% CI 1.1–10.4; p = 0.033). Conclusions In this large cohort study, VAC therapy in patients with disseminated FG may offer an advantage in terms of 10-weeks wound closure cumulative rate and OS at 90 days after initial surgery.

Purpose The Ubiquitin Proteasome System (UPS) is a selective degradation system mediating the removal of intracellular unfolded/misfolded proteins and is essential for cardiomyocyte (CM) health. Substrate specificity and ubiquitination rate are mediated by E3 ubiquitin-ligases, such as Atrogin1 and MuRF1, which are specifically expressed in muscle cells. Perturbation of protein quality control causes aggregation of misfolded proteins, leading to CM proteotoxicity. UPS dysfunction occurs in ageing, a risk factor for cardiac hypertrophy and HF. We recently demonstrated that Atrogin-1 is essential to maintain CM health, during ageing. Whether MuRF1 plays similar roles in heart adaptation to ageing is still unexplored, and different studies have yielded contrasting results. Methods To assess the role of MuRF1 in heart homeostasis, we combined echocardiography, histology, IF, TUNEL assay and EM on heart sections from MuRF1 knock-out (KO) mice, and littermate controls, at 3, 10 and 24 mo. RTqPCR and WB assessed markers of UPS and extracellular matrix. Langendorff procedure was used to separate CMs from cardiac fibroblasts. Molecular and IF analyses were performed in heart samples from patients affected by aortic stenosis. Results MuRF1 ablation leads to cardiac hypertrophy, progressing during ageing (LV CM areas: 3 mo., KO: 329.14±10.66 vs Ctrl: 296.25±5.43; 10 mo., KO: 399.73±7.64 vs Ctrl: 247.49±3.67; 24 mo., KO: 418.89±11.10 vs. Ctrl: 209.93±4.48, in μm2). The hypertrophic remodeling was accompanied by diastolic dysfunction in the adulthood and, during ageing, also by systolic dysfunction (EF, 24 mo., KO: 29.55±8.82 vs Ctrl: 51.23±6.56, in %). Loss of MuRF1 causes increased interstitial collagen -I and -VI deposition, even before the onset of contractile dysfunction, followed by activation of Matrix MetalloProteinases (MMPs), suggesting that such alterations may be responsible for decreased cardiac performance. Interestingly, collagen established rings enveloping MuRF1 KO CMs and such fibrotic remodeling was not accompanied by increased CM apoptosis, nor myofibroblast activation. Such peculiar remodelling, called peri-endomysial fibrosis, was detected in hearts from patients with aortic stenosis, a condition in which MuRF1 levels decrease. Our data supports that MuRF1 has a role in CM-dependent regulation of the extracellular matrix (ECM) dynamics. Consistently, MuRF1 downregulation in normal cultured CMs demonstrated that such ubiquitin ligase impacts on signaling pathways involved in the control of the ECM homeostasis. In addition, fibroblasts treated with culture medium conditioned by MuRF1 KO CM display increased collagen and MMP expression. Conclusions We identifieded a novel role of MuRF1 in the control of CM proteostasis, and unveiled that in addition to cardiac fibroblasts, CM may directly regulate ECM dynamics, indicating that the correct function of MuRF1 is essential for heart adaptation to aging. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): University of Padova

Introduction: COVID-19 outbreak represented an unprecedented event that led to a redefinition of health care systems worldwide The impact of the emergency required a deviation of the care toward the assistance to COVID-19 patients, with reduction of resources for elective activities, including surgery We aim to report the decrease of urological surgical activity during the first weeks from the beginning of the pandemic, aiming to highlight the prioritization we applied to select patients for surgery Materials and methods: Thirty-three urological units with physicians affiliated to the AGILE group were involved in a survey Urologists were asked to report the amount of surgical elective procedures week- by-week, from the beginning of the emergency to the following month The type of surgery (oncologic, for urolithiasis, for benign prostate obstruction, other) was assessed as well Results: The 33 hospitals involved in the study account, globally, for 22,945 beds and are distributed in 13/20 Italian regions Before the outbreak, the involved urology units performed an overall amount of 1,213 procedures per week, half of which were oncological By the 20 of March, the amount of surgery declined by 78% Lombardy, the first region with positive-cases, experienced a 94% reduction The decrease in oncological and non- oncological surgical activity was 35,9% and 89%, respectively Among non-oncological procedures, stone surgery declined by 35,9% as well, whereas BPH and minor urological procedures completely dropped Reassessing for surgical activity on 20, April, a slight trend toward surgical restoration (+11%) started to appear Conclusions: Italy, the country with the highest fatality rate from COVID-19, had experienced a sudden decline in surgical activity;by the end of April, a current trend toward restoration of surgery started to appear Criteria for prioritization were consistent with an urgent/ emergent principle, with trauma, tumours and septic conditions being the ones prioritized The Italian experience can be helpful for future surgical pre-planning in other countries or pandemic settings Smart Communications (SC1–SC28) Andrology

Background: VOCs originate from metabolic pathophysiological processes at respiratory and systemic level. VOCs pattern has previously been analysed with E-nose on exhaled air; in CF it distinguished P.aeruginosa and A.fumigatus colonization. Aim: to evaluate exhaled VOCs pattern with E-nose (Cyranose320®)in HC and in CF pts; to compare exhaled VOCs in stable CF pts (StCF) and in pts with respiratory exacerbation (ExCF). Methods: a cross-sectional experimental study was approved by Verona Ethic Committee. Spirometry, N2multiple breath washout (MBW) and exhaled air collection were performed in 29 CF pts (age 12.56 ± 4.08; 23 StCF, 6 ExCF) and 15 HC (age 13.36 ±4.13). VOCs analysis was performed with Cyranose320®. The output data underwent multivariate analysis and Principal Component Analysis (PCA). Results: A number of E-nose sensors (S6-8, S10, S12, S21, S27, S28) significantly distinguished between HC and CF pts; also, StCF could be discriminated from ExCf (S6-8, S10, S12, S21, S27). For sensors distinguishing between HC and CF pts, AUC of ROC was more than accurate for S12 (0.85), good for S6-7, S21, S27 (0.73-0.76). PCA identified PC10, PC1, PC7 and PC2, which proved the best PC in predicting between HC and CF subsets. Spirometry distinguished HC from ExCF, but not from St CF, whereas N2MBW distinguished also StCF from HC. Conclusion: E-nose analysis of exhaled air samples distinguished HC from CF pts, and among CF pts, StCF from ExCF. N2MBW distinguished all 3 subsets, whereas spirometry only distinguished ExCF from HC. E-nose is a promising tool for clinical and research setting.

Aging is associated with alterations in skeletal muscle autophagy, potentially affecting both muscle mass and quality in a negative manner. Strength training with protein supplementation has been reported to improve both muscle mass and quality in frail elderly individuals, but whether improvements are accompanied by alterations in protein quality control is not known. To address this issue, we investigated protein degradation markers in skeletal muscle biopsies (m. vastus lateralis) from twenty-four frail elderly men and women (86 ± 7 yr) after acute and chronic (10 weeks) strength training with protein supplementation (ST + PRO) or protein supplementation alone (PRO). Acute increases in mRNA expression of genes related to the ubiquitin proteasome system (MuRF-1, MUSA1), autophagy (ATG7, LC3, p62), and mitochondrial fission (DRP1) were observed after the first, but not after the last training session in ST + PRO. Acute changes in gene expression were accompanied by changes in protein levels of both LC3-I and LC3-II. Hence, the acute training-induced activation of proteasomal degradation and autophagy seems to depend on training status, with activation in the untrained, but not trained state. The ten-week training intervention did not affect basal levels of autophagy mRNAs and proteins, and neither markers of the ubiquitin-proteasome system. This suggests that a relatively short period of strength training may not be sufficient to increase the basal rate of protein degradation in frail elderly.

The loss of skeletal muscle mass under a wide range of acute and chronic maladies is associated with poor prognosis, reduced quality of life, and increased mortality. Decades of research indicate the importance of skeletal muscle for whole body metabolism, glucose homeostasis, as well as overall health and wellbeing. This tissue’s remarkable ability to rapidly and effectively adapt to changing environmental cues is a double-edged sword. Physiological adaptations that are beneficial throughout life become maladaptive during atrophic conditions. The atrophic program can be activated by mechanical, oxidative, and energetic distress, and is influenced by the availability of nutrients, growth factors, and cytokines. Largely governed by a transcription-dependent mechanism, this program impinges on multiple protein networks including various organelles as well as biosynthetic and quality control systems. Although modulating muscle function to prevent and treat disease is an enticing concept that has intrigued research teams for decades, a lack of thorough understanding of the molecular mechanisms and signaling pathways that control muscle mass, in addition to poor transferability of findings from rodents to humans, has obstructed efforts to develop effective treatments. Here, we review the progress made in unraveling the molecular mechanisms responsible for the regulation of muscle mass, as this continues to be an intensive area of research.

Objectives To report the frequency and characteristics of growth hormone (GH) deficiency (GHD) in adolescents who had normalized GH secretion at mid-puberty and to identify possible factors predictive for GH sufficiency at puberty. Design Clinical analysis of children affected by GHD at five time points: diagnosis; first year of therapy; intermediate stage of puberty; retesting and end of growth phase. Methods The study population was 80 children with idiopathic GHD and treated with GH for at least 2 years. Treatment was discontinued at the intermediate stage of puberty. Retesting with an arginine test was performed 12 weeks later. If GH peak at retesting was ≥8 μg/L, the therapy was definitively discontinued, otherwise it was restarted and continued until achievement of near-final height. Results GH therapy was discontinued in 44 children (55%), and restarted in 36 (45%). No evidence of differences in definitive height and in the delta height between the genetic target and the definitive height was found between the two groups. The only predictive factor for GHD at mid-puberty was the insulin growth factor-1 (IGF-1) level at 1 year of GH treatment. Conclusions GH secretion should be retested at mid-puberty. Retesting at puberty may reduce potential side effects and minimize costs, without impairing growth potential and final height.

Background: The ERG1a potassium channel has been detected in the atrophying skeletal muscle of mice experiencing either muscle disuse or cancer cachexia and further evidenced to contribute to muscle deterioration by enhancing ubiquitin proteolysis; however, to our knowledge, ERG1 has not been reported in human skeletal muscle. Methods and Results: Here, using immunohistochemistry, we detect ERG1 immunofluorescence in human Rectus abdominis skeletal muscle sarcolemma. Further, using single point brightness data, we report detection of ERG1 immunofluorescence at low levels in the Rectus abdominis muscle sarcolemma of young adult humans and show that it trends toward greater levels (10.6%) in healthy aged adults. Interestingly, we detect ERG1 immunofluorescence at a statistically greater level (53.6%; p

PurposeThe determinants of physical fitness in children have been given limited attention. In particular, the relative role of chronological age, sex, anthropometry, and body composition in physical fitness of children has been barely investigated. This cross-sectional study investigated determinants of physical fitness using a set of predictive variables including, in addition to chronological age and sex, a large panel of anthropometric measurements as well as body composition. The study was carried out in a convenience sample of children aged 6–12 participating in a summer camp.MethodsOne-hundred-ninety-three children (128 males) fulfilled all requirements and entered analysis. Health-related physical fitness components (speed, muscular power and balance) were explored by means of field tests, namely the 30-m dash test for running speed, the standing long jump and the seated chest pass test for lower limbs and upper body muscular power, respectively, and the flamingo balance test for static balance. Determinants of physical fitness were investigated by regression analysis using chronological age, sex, anthropometry, and body composition in a hierarchical approach. To minimize the expected effect of collinearity in predictor variables, an original statistical approach using Random Forests analysis was adopted.ResultsAge predicted 45.2%, 43.6%, 35.6% and 25.6%; and sex 9.5%, 10.7%, 6.3% and 2.0% of variance in the 30-m dash, seated chest pass, standing long jump, and flamingo balance test, respectively. Anthropometry and body composition explained a limited or no percentage of variance. The adjustedR2(root mean square error) was 0.61 (0.31 s), 0.45 (0.32 m), 0.58 (0.15 m) and 0.41 (0.75 logs) for the 30-m dash, seated chest pass, standing long jump, and flamingo balance test, respectively making these models useful when physical fitness tests are not feasible.ConclusionsWe highlighted the respective role of chronological age, sex, anthropometry, and body composition in physical fitness of children in the wide age range 6–12 years. Data confirm and expand on previous literature by showing with a strictly conservative statistical approach that chronological age is a main determinant of physical fitness of both boys and girls, sex playing a limited role. The role of anthropometry was even less important, and no role was found for body composition. These findings should be considered when planning/implementing motor development or physical education programs.

Muscular Dystrophies are severe genetic diseases due to mutations in structural genes, characterized by progressive muscle wasting that compromises patients’ mobility and respiratory functions. Literature underlined oxidative stress and inflammation as key drivers of these pathologies. Interestingly among different myofiber classes, type I fibers display a milder dystrophic phenotype showing increased oxidative metabolism. This work shows the benefits of a cyanidin-enriched diet, that promotes muscle fiber-type switch and reduced inflammation in dystrophic alpha-sarcoglyan (Sgca) null mice having, as a net outcome, morphological and functional rescue. Notably, this benefit is achieved also when the diet is administered in dystrophic animals when the signs of the disease are seriously evident. Our work provides compelling evidence that a cyanidin-rich diet strongly delays the progression of muscular dystrophies, paving the way for a combinatorial approach where nutritional-based reduction of muscle inflammation and oxidative stress facilitate the successful perspectives of definitive treatments.

To discover patterns in data is about using Data Science to reveal the hidden mechanisms governing the analyzed phenomena. Data Science is able to uncover lots of different patterns such as distributions, associations, similarities, interactions, classifications and trends, that can be described, measured and modelled. There are only patterns, patterns on top of patterns, and patterns that affect other patterns. This chapter discusses some examples of patterns that can be revealed and many statistical tools that can be exploited to analyze data. It is concerned with the identification of the factors determining high-pressure game situations and affecting the scoring probability. An important issue for the analysis of games is to determine the frequency of occurrence of some events with respect to some other concurrent variable. A broad set of extremely powerful tools to discover hidden structures and patterns in data comes from machine learning techniques that make a large number of algorithms available for both supervised and unsupervised learning.

Data Mining algorithms try to address the issue as scientifically as possible, defining methods and rules aimed to assign units of observation into classes, which are not defined a priori, and are supposed to somehow reflect the structure of the entities that the data represent. Homogeneity within each cluster and the degree of separation among clusters are measured by referring to a distance metric or a dissimilarity measure. Cluster Analysis is a classification technique aiming at dividing individual cases into groups such that the cases in a cluster are very similar to one another and very different from the cases in other clusters. The ordering of pairs of units is sensitive to the selected measure of distance or dissimilarity, so an important step in Cluster Analysis is the choice of the most appropriate measure, which in practice should be based on the type of data at hand: for numerical variables while for categorical variables dissimilarity measures should be preferred.

Although we have shown that catecholamines suppress the activity of the Ubiquitin-Proteasome System (UPS) and atrophy-related genes expression through a cAMP-dependent manner in skeletal muscle from rodents, the underlying mechanisms remain unclear. Here, we report that a single injection of norepinephrine (NE; 1 mg kg-1 ; s.c) attenuated the fasting-induced up-regulation of FoxO-target genes in tibialis anterior (TA) muscles by the stimulation of PKA/CREB and Akt/FoxO1 signaling pathways. In addition, muscle-specific activation of PKA by the overexpression of PKA catalytic subunit (PKAcat) suppressed FoxO reporter activity induced by (1) a wild-type; (2) a non-phosphorylatable; (3) a non-phosphorylatable and non-acetylatable forms of FoxO1 and FoxO3; (4) downregulation of FoxO protein content, and probably by (5) PGC-1α up-regulation. Consistently, the overexpression of the PKAcat inhibitor (PKI) up-regulated FoxO activity and the content of Atrogin-1 and MuRF1, as well as induced muscle fiber atrophy, the latter effect being prevented by the overexpression of a dominant negative (d. n.) form of FoxO (d.n.FoxO). The sustained overexpression of PKAcat induced fiber-type transition toward a smaller, slower, and more oxidative phenotype and improved muscle resistance to fatigue. Taken together, our data provide the first evidence that endogenous PKA activity is required to restrain the basal activity of FoxO and physiologically important to maintain skeletal muscle mass.

© The Author(s) 2020. Open Access: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/., A unique property of skeletal muscle is its ability to adapt its mass to changes in activity. Inactivity, as in disuse or aging, causes atrophy, the loss of muscle mass and strength, leading to physical incapacity and poor quality of life. Here, through a combination of transcriptomics and transgenesis, we identify sestrins, a family of stress-inducible metabolic regulators, as protective factors against muscle wasting. Sestrin expression decreases during inactivity and its genetic deficiency exacerbates muscle wasting; conversely, sestrin overexpression suffices to prevent atrophy. This protection occurs through mTORC1 inhibition, which upregulates autophagy, and AKT activation, which in turn inhibits FoxO-regulated ubiquitin-proteasome-mediated proteolysis. This study reveals sestrin as a central integrator of anabolic and degradative pathways preventing muscle wasting. Since sestrin also protected muscles against aging-induced atrophy, our findings have implications for sarcopenia., The authors acknowledge funding from the Spanish Ministry of Science, Innovation and Universities, Spain (grants SAF2015-67369-R, RTI2018-096068-B-I00, and SAF 2015-70270-REDT, a María de Maeztu Unit of Excellence award to UPF [MDM-2014-0370], and the UPF-CNIC collaboration agreement, ERC-2016-AdG-741966, La Caixa-HEALTH (HR17-00040), MDA, UPGRADE-H2020-825825, AFM and DPP-E. The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the Ministerio de Ciencia, Innovación y Universidades (MCNU) and the Pro CNIC Foundation, and is a Severo Ochoa Center of Excellence (SEV-2015-0505). Work was also supported by the Ellison Medical Foundation (AG-SS-2440-10 to M.K. and AG-NS-0932-12 to J.H.L.), NIH (R01DK114131, R01DK111465, and R01DK102850 to J.H.L.), CARIPARO and H2020-MSCA-RISE-2014 (to M.S.), and the Russian Science Foundation (Grant 17-14-01420 to A.V.B.). J.S. acknowledges funding from a Juan de La Cierva Postdoctoral Fellowship.

Polyglutamine (polyQ) expansions in the androgen receptor (AR) gene cause spinal and bulbar muscular atrophy (SBMA), a neuromuscular disease characterized by lower motor neuron (MN) loss and skeletal muscle atrophy, with an unknown mechanism. We generated new mouse models of SBMA for constitutive and inducible expression of mutant AR and performed biochemical, histological and functional analyses of phenotype. We show that polyQ-expanded AR causes motor dysfunction, premature death, IIb-to-IIa/IIx fiber-type change, glycolytic-to-oxidative fiber-type switching, upregulation of atrogenes and autophagy genes and mitochondrial dysfunction in skeletal muscle, together with signs of muscle denervation at late stage of disease. PolyQ expansions in the AR resulted in nuclear enrichment. Within the nucleus, mutant AR formed 2% sodium dodecyl sulfate (SDS)-resistant aggregates and inclusion bodies in myofibers, but not spinal cord and brainstem, in a process exacerbated by age and sex. Finally, we found that two-week induction of expression of polyQ-expanded AR in adult mice was sufficient to cause premature death, body weight loss and muscle atrophy, but not aggregation, metabolic alterations, motor coordination and fiber-type switch, indicating that expression of the disease protein in the adulthood is sufficient to recapitulate several, but not all SBMA manifestations in mice. These results imply that chronic expression of polyQ-expanded AR, i.e. during development and prepuberty, is key to induce the full SBMA muscle pathology observed in patients. Our data support a model whereby chronic expression of polyQ-expanded AR triggers muscle atrophy through toxic (neomorphic) gain of function mechanisms distinct from normal (hypermorphic) gain of function mechanisms.

Muscle regeneration is sustained by infiltrating macrophages and the consequent activation of satellite cells1–4. Macrophages and satellite cells communicate in different ways1–5, but their metabolic interplay has not been investigated. Here we show, in a mouse model, that muscle injuries and ageing are characterized by intra-tissue restrictions of glutamine. Low levels of glutamine endow macrophages with the metabolic ability to secrete glutamine via enhanced glutamine synthetase (GS) activity, at the expense of glutamine oxidation mediated by glutamate dehydrogenase 1 (GLUD1). Glud1-knockout macrophages display constitutively high GS activity, which prevents glutamine shortages. The uptake of macrophage-derived glutamine by satellite cells through the glutamine transporter SLC1A5 activates mTOR and promotes the proliferation and differentiation of satellite cells. Consequently, macrophage-specific deletion or pharmacological inhibition of GLUD1 improves muscle regeneration and functional recovery in response to acute injury, ischaemia or ageing. Conversely, SLC1A5 blockade in satellite cells or GS inactivation in macrophages negatively affects satellite cell functions and muscle regeneration. These results highlight the metabolic crosstalk between satellite cells and macrophages, in which macrophage-derived glutamine sustains the functions of satellite cells. Thus, the targeting of GLUD1 may offer therapeutic opportunities for the regeneration of injured or aged muscles. Mouse models of muscle injuries and ageing characterized by low levels of intra-tissue glutamine are ameliorated by macrophage-specific deletion or systemic pharmacological inhibition of glutamate dehydrogenase 1, which results in constitutively high activity of glutamine synthetase.

To date there is no anthropometric equation specific to athletes with unilateral lower limb amputation to estimate the percentage of fat mass (%FM). This study investigated the accuracy of a set of anthropometric equations validated on able-bodied populations to predict the %FM assessed by-means of dual-energy x-ray absorptiometry (DXA) in athletes with unilateral lower limb amputation. Furthermore, a predictive anthropometric equation specific to athletes with unilateral lower limb amputation was developed from skinfold thickness measurements using DXA as the reference method for the estimation of the %FM. Twenty-nine white male athletes with unilateral lower limb amputation underwent a DXA scan and an anthropometric assessment on the same day. The %FM, calculated through several existing anthropometric equations validated upon able-bodied populations, was compared with the DXA-measured %FM (%FM_DXA). Accuracy and agreement between the two methods was computed with two-tailed paired-sample t-test, concordance correlation coefficient, reduced major axis regression and Bland-Altman analysis. A stepwise multiple regression analysis with the %FM_DXA as the dependent variable and age and nine skinfold thicknesses as potential predictors was carried out and validated using a repeated 10-fold cross-validation. A linear regression analysis with the sum of nine skinfolds as the independent variable was also carried out and validated using a repeated 10-fold cross-validation. The results showed that the anthropometric equations validated on able-bodied populations are inaccurate in the estimation of %FM_DXA with an average bias ranging from 0.51 to −13.70%. Proportional bias was also found revealing that most of the anthropometric equations considered, tended to underestimate/overestimate the %FM_DXA as body fat increased. Regression analysis produced two statistically significant models (P < 0.001 for both) which were able to predict more than 93% of total variance of %FM_DXA from the values of four skinfold measurements (i.e., thigh, abdominal, subscapular and axillary skinfold measurements) or from the sum of 9 skinfolds. Repeated cross-validation analysis highlighted a good predictive performance of the proposed equations. The predictive equations proposed in this study represent a useful tool for clinicians, nutritionists, and physical conditioners to evaluate the physical and nutritional status of athletes with unilateral lower limb amputation directly in the field.

Background A microscopic analysis of tissue is the gold standard for cancer detection. Hematoxylin-eosin (HE) for the reporting of prostate biopsy (PB) is conventionally based on fixation, processing, acquisition of glass slides, and analysis with an analog microscope by a local pathologist. Digitalization and real-time remote access to images could enhance the reporting process, and form the basis of artificial intelligence and machine learning. Fluorescence confocal microscopy (FCM), a novel optical technology, enables immediate digital image acquisition in an almost HE-like resolution without requiring conventional processing. Objective The aim of this study is to assess the diagnostic ability of FCM for prostate cancer (PCa) identification and grading from PB. Design, setting, and participants This is a prospective, comparative study evaluating FCM and HE for prostate tissue interpretation. PBs were performed (March to June 2019) at a single coordinating unit on consecutive patients with clinical and laboratory indications for assessment. FCM digital images (n = 427) were acquired immediately from PBs (from 54 patients) and stored; corresponding glass slides (n = 427) undergoing the conventional HE processing were digitalized and stored as well. A panel of four international pathologists with diverse background participated in the study and was asked to evaluate all images. The pathologists had no FCM expertise and were blinded to clinical data, HE interpretation, and each other’s evaluation. All images, FCM and corresponding HE, were assessed for the presence or absence of cancer tissue and cancer grading, when appropriate. Reporting was gathered via a dedicated web platform. Outcome measurements and statistical analysis The primary endpoint is to evaluate the ability of FCM to identify cancer tissue in PB cores (per-slice analysis). FCM outcomes are interpreted by agreement level with HE (K value). Additionally, either FCM or HE outcomes are assessed with interobserver agreement for cancer detection (presence vs absence of cancer) and for the discrimination between International Society of Urologic Pathologists (ISUP) grade = 1 and ISUP grade > 1 (secondary endpoint). Results and limitations Overall, 854 images were evaluated from each pathologist. PCa detection of FCM was almost perfectly aligned with HE final reports (95.1% of correct diagnosis with FCM, κ = 0.84). Inter-rater agreement between pathologists was almost perfect for both HE and FCM for PCa detection (0.98 for HE, κ = 0.95; 0.95 for FCM, κ = 0.86); for cancer grade attribution, only a moderate agreement was reached for both HE and FCM (HE, κ = 0.47; FCM, κ = 0.49). Conclusions FCM provides a microscopic, immediate, and seemingly reliable diagnosis for PCa. The real-time acquisition of digital images—without requiring conventional processing—offers opportunities for immediate sharing and reporting. FCM is a promising tool for improvements in cancer diagnostic pathways. Patient summary Fluorescence confocal microscopy may provide an immediate, microscopic, and apparently reliable diagnosis of prostate cancer on prostate biopsy, overcoming the standard turnaround time of conventional processing and interpretation.

"Several research studies were focused to understand how grapevine cultivars respond to environment; nevertheless, the biological mechanisms tuning this phenomenon need to be further deepened. Particularly, the molecular processes underlying the interplay between clones of the same cultivar and environment were poorly investigated. To address this issue, we analyzed the transcriptome of berries from three ""Nebbiolo"" clones grown in different vineyards, during two ripening seasons. RNA-sequencing data were implemented with analyses of candidate genes, secondary metabolites, and agronomical parameters. This multidisciplinary approach helped to dissect the complexity of clone × environment interactions, by identifying the molecular responses controlled by genotype, vineyard, phenological phase, or a combination of these factors. Transcripts associated to sugar signalling, anthocyanin biosynthesis, and transport were differently modulated among clones, according to changes in berry agronomical features. Conversely, genes involved in defense response, such as stilbene synthase genes, were significantly affected by vineyard, consistently with stilbenoid accumulation. Thus, besides at the cultivar level, clone-specific molecular responses also contribute to shape the agronomic features of grapes in different environments. This reveals a further level of complexity in the regulation of genotype × environment interactions that has to be considered for orienting viticultural practices aimed at enhancing the quality of grape productions."

OBJECTIVE To establish the relationship between vitamin D serum levels, pulmonary function, asthma control, and passive smoking exposure in children with asthma. METHODS We studied the relationship between 25-hydroxy cholecalciferol (25[OH]D) concentrations and baseline spirometry and levels of asthma control, and the effect of parental tobacco smoke exposure in 152 white children (84 boys [55.3%]) with a mean age ± standard deviation of 9.9 ± 2.0 years (range 5-15 years) in a cross-sectional study carried out during the winter and early spring. RESULTS Only 9.9% of our children had a sufficient serum 25(OH)D level (at least 30-40 ng/mL). A significant positive correlation was found between the force vital capacity % predicted, forced expiratory volume in the first second of expiration % predicted, and serum 25(OH)D level (r = 0.36, p < 0.001 for both). The subjects with controlled asthma had higher serum levels of 25(OH)D than children with partially controlled or noncontrolled asthma, both according to Global Initiative for Asthma parameters and the Test for the control of asthma in childhood (p = 0.011). Children with both nonsmoking parents presented significantly higher serum levels of 25(OH)D than children with both smoking parents (median, 20.5 ng/mL [interquartile range {IQR}, 16.6-24.0 ng/mL] versus median, 14.5 ng/mL [IQR, 11.1-19.1 ng/mL], respectively; p < 0.001), with intermediate values for children exposed to single maternal (median, 20.3 ng/mL [IQR, 13.0-23.2 ng/mL]) or to paternal smoking (median, 17.8 ng/mL [IQR, 14.7-22.1 ng/mL]). CONCLUSION Our results indicated that hypovitaminosis D was frequent in children with asthma who lived in a Mediterranean country. In these children, lower levels of vitamin D were associated with reduced asthma control and passive smoking exposure.

Local recurrences after breast conserving treatment are mainly close to the original tumor site, and as such shorter fractionation strategies focused on and nearest mammary gland, i.e. accelerated partial breast irradiation (APBI), have been developed. Stereotactic APBI has been attempted, although there is little experience using CyberKnife (CK) for early breast cancer. This pilot study was designed to assess the feasibility of CK-APBI on 20 evaluable patients of 29 eligible, followed for 2 years. The primary endpoint was acute/sub-acute toxicity; secondary endpoints were late toxicity and the cosmetic result. Mean pathological tumor size was 10.5 mm (±4.3, range 3–18), 8 of these patients were classified as LumA-like, 11 as LumB-like, and 1 as LumB-HER2-enriched. Using CK-APBI with Iris, the treatment time was approximately 60 min (range~ 35 to ~ 120). All patients received 30 Gy in five fractions delivered to the PTV. The median number of beams was 180 (IQR 107–213; range:56–325) with a median PTV isodose prescription of 86.0% (IQR 85.0–88.5; range:82–94). The median PTV was 88.1 cm3 (IQR 63.8–108.6; range:32.3–238.8). The median breast V100 and V50 was 0.6 (IQR 0.1–1.5; range:0–13) and 18.6 (IQR 13.1–21.7; range:7.5–37), respectively. The median PTV minimum dose was 26.2 Gy (IQR 24.7–27.6; range 22.3–29.3). Mild side effects were recorded during the period of observation. Cosmetic evaluations were performed by three observers from the start of radiotherapy up to 2 years. Patients’ evaluation progressively increase from 60% to 85% of excellent rating; this trend was similar to that of external observer. These preliminary results showed the safe feasibility of CK-APBI in early breast cancer, with mild acute and late toxicity and very good cosmetic results. The present study is registered at Clinicaltrial.gov ( NCT02896322 ). Retrospectively egistered August 4, 2016.

Mechanical ventilation (MV) results in the rapid development of ventilator-induced diaphragm dysfunction (VIDD). While the mechanisms responsible for VIDD are not fully understood, recent data reveal that prolonged MV activates autophagy in the diaphragm, which may occur as a result of increased cellular reactive oxygen species (ROS) production. Therefore, we tested the hypothesis that (1) accelerated autophagy is a key contributor to VIDD; and that (2) oxidative stress is required to increase the expression of autophagy genes in the diaphragm. Our findings reveal that targeted inhibition of autophagy in the rat diaphragm prevented MV-induced muscle atrophy and contractile dysfunction. Attenuation of VIDD in these animals occurred as a result of increased diaphragm concentration of the antioxidant catalase and reduced mitochondrial ROS emission, which corresponded to reductions in the activity of calpain and caspase-3. To determine if increased ROS production is required for the upregulation of autophagy biomarkers in the diaphragm, rats that were administered the mitochondrial-targeted peptide SS-31 during MV. Results from this study demonstrated that mitochondrial ROS production in the diaphragm during MV is required for the increased expression of key autophagy genes (i.e. LC3, Atg7, Atg12, Beclin1 and p62), as well as for increased activity of cathepsin L. Together, these data reveal that autophagy is required for VIDD, and that autophagy inhibition reduces MV-induced diaphragm ROS production and prevents a positive feedback loop whereby increased autophagy is stimulated by oxidative stress, resulting in further increases in ROS and autophagy.

In this paper, we analyze the shooting performance of basketball players by examining the factors that may generate high-pressure game situations. Using play-by-play data from the Italian “Serie A2” Championship 2015/2016 to build the model, we validate the main results using data from the Olympic Basketball Tournament “Rio 2016” to determine whether the relationships we identified can be confirmed using data from players at a very different professional level. After a preliminary exploratory analysis, we (1) develop a multivariate model based on the Classification and Regression Tree algorithm in order to investigate how selected high-pressure situations, jointly considered, affect scoring probability and then propose new shooting performance measures; (2) investigate players’ personal reactions to selected high-pressure game situations by introducing additional new measures, improving the indices currently used to measure shooting performance. The results are interesting and easy to interpret with the aid of some insightful graphical representations. Our approach can be exploited by both scouts and coaches to understand important player characteristics and, ultimately, to measure and enhance a team’s performance.

Introduction Coagulation plays a crucial role in coronary artery disease (CAD) contributing to both atherosclerotic plaque development and acute thrombotic complications, like myocardial infarction (MI). Coagulation biomarkers have been linked with ischemic heart disease, but results are still controversial. Materials and methods D-dimer and thrombin generation, two “overall” coagulation assays, were evaluated in 775 subjects with or without angiographically-proven CAD (170 CAD-free and 605 CAD, 355 of whom with history of previous MI). Subjects taking anticoagulant drugs or with any acute illness were excluded. D-dimer plasma concentration was determined by an immuno-turbidimetric assay. Thrombin generation was assessed as the ability of plasma to generate thrombin triggered by the addition of tissue factor ex-vivo by means of a chromogenic method. Results Both D-dimer and thrombin generation parameters were associated with several traditional cardiovascular risk factors. Lag-time, time-to-peak, peak height, and Endogenous Thrombin Potential (ETP), as well as D-dimer levels, were higher in CAD patients than in CAD-free subjects. After adjustment for all the traditional risk factors, only ETP levels remained significantly associated with CAD (the highest versus the lowest tertile: OR 2.61 with 95%CI 1.14–5.99), but without improvement of C-statistic. The association of D-dimer vanished after adjustment for inflammatory markers. No difference of either D-dimer or thrombin generation parameters was found between CAD patients with or without previous MI history. Conclusions Our results suggest that an increased plasma thrombin potential is characteristic in patients with clinically stable CAD, irrespective of previous MI history and independent of traditional cardiovascular risk factors.

Grapevine (Vitis vinifera) berry development involves a succession of physiological and biochemical changes reflecting the transcriptional modulation of thousands of genes. Although recent studies have investigated the dynamic transcriptome during berry development, most have focused on a single grapevine variety, so there is a lack of comparative data representing different cultivars. Here, we report, to our knowledge, the first genome-wide transcriptional analysis of 120 RNA samples corresponding to 10 Italian grapevine varieties collected at four growth stages. The 10 varieties, representing five red-skinned and five white-skinned berries, were all cultivated in the same experimental vineyard to reduce environmental variability. The comparison of transcriptional changes during berry formation and ripening allowed us to determine the transcriptomic traits common to all varieties, thus defining the core transcriptome of berry development, as well as the transcriptional dynamics underlying differences between red and white berry varieties. A greater variation among the red cultivars than between red and white cultivars at the transcriptome level was revealed, suggesting that anthocyanin accumulation during berry maturation has a direct impact on the transcriptomic regulation of multiple biological processes. The expression of genes related to phenylpropanoid/flavonoid biosynthesis clearly distinguished the behavior of red and white berry genotypes during ripening but also reflected the differential accumulation of anthocyanins in the red berries, indicating some form of cross talk between the activation of stilbene biosynthesis and the accumulation of anthocyanins in ripening berries.