Recombinant OC43 SARS-CoV-2 spike replacement virus: An improved BSL-2 proxy virus for SARS-CoV-2 neutralization assays.

We generated a replication-competent OC43 human seasonal coronavirus (CoV) expressing the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike in place of the native spike (rOC43-CoV2 S). This virus is highly attenuated relative to OC43 and SARS-CoV-2 in cultured cells and animals and is classified as a biosafety level 2 (BSL-2) agent by the NIH biosafety committee. Neutralization of rOC43-CoV2 S and SARS-CoV-2 by S-specific monoclonal antibodies and human sera is highly correlated, unlike recombinant vesicular stomatitis virus-CoV2 S. Single-dose immunization with rOC43-CoV2 S generates high levels of neutralizing antibodies against SARS-CoV-2 and fully protects human ACE2 transgenic mice from SARS-CoV-2 lethal challenge, despite nondetectable replication in respiratory and nonrespiratory organs. rOC43-CoV2 S induces S-specific serum and airway mucosal immunoglobulin A and IgG responses in rhesus macaques. rOC43-CoV2 S has enormous value as a BSL-2 agent to measure S-specific antibodies in the context of a bona fide CoV and is a candidate live attenuated SARS-CoV-2 mucosal vaccine that preferentially replicates in the upper airway.

Preliminary Application of Infrared Thermography to Monitoring of Skin Temperature Asymmetries in Professional Padel Players.

The aim of the present study was to evaluate skin temperature (Tsk) asymmetries, using infrared thermography, in professional padel players before (PRE), after (POST) and 10 min after training (POST10), and their relationship with perceptual variables and training characteristics. Thermal images were taken of 10 players before, after and 10 min after a standardized technical training. After training, Tsk of the dominant side was higher than before training in the anterior forearm (30.8 ± 0.4 °C vs. 29.1 ± 1.2 °C, p < 0.01; ES = 1.9), anterior shoulder (31.6 ± 0.6 °C vs. 30.9 ± 0.6 °C, p < 0.05; ES = 1.0) posterior arm (29.5 ± 1.0 °C vs. 28.3 ± 1.2 °C, p < 0.05; ES = 1.0), and posterior forearm (30.8 ± 0.9 °C vs. 29.3 ± 1.6 °C, p < 0.05; ES = 1.1). Likewise, these differences were significant POST10 in the anterior arm, anterior forearm, anterior shoulder, posterior arm and posterior forearm. Comparing the different moments of measurement (PRE, POST and POST10), the temperature was higher POST10 in all the regions analyzed except for the shoulder, abdominals, and lower back. Also, correlations were found between fatigue variation and temperature variation between limbs (Tsk dominance), and no correlation was found except between age and posterior thigh (|r| = 0.69; p < 0.05), and between the racket mass and anterior knee (|r| = 0.81; p < 0.01). In conclusion, infrared thermography allows monitoring of skin asymmetries between limbs in professional padel players, but these asymmetries were not related to overall fatigue variation, overall pain variation, years of experience and training hours.

Early poststroke clinically significant fatigue predicts functional independence: a prospective longitudinal study.

Background: Poststroke fatigue is a prevalent issue among stroke survivors, significantly impeding functional recovery and diminishing their quality of life. Aim: This prospective cohort study aims to investigate the association between poststroke fatigue and the extent of functional recovery in survivors of ischemic and hemorrhagic strokes. Additionally, it seeks to delineate the temporal progression of poststroke fatigue in these two stroke subtypes. Methods: We assessed a cohort of 79 patients recovering from acute ischemic or hemorrhagic strokes. Poststroke fatigue was quantified using the Fatigue Severity Scale (FSS) and the Numeric Rating Scale (NRSfatigue). Patients' condition was evaluated using the National Institute of Health Stroke Scale (NIHSS), and functional independence levels were determined using the Barthel Index for Activities of Daily Living (BIADL) and the Modified Rankin Scale (MRS). Depressive mood and pain were measured using the Beck Depression Inventory (BDI) and the Numeric Rating Scale for pain (NRSpain), respectively. Results: Our primary findings indicate that the early manifestation of clinically significant fatigue (CSF) is predictive of a poorer trajectory in functional independence levels during recovery. Furthermore, we observed differing patterns of fatigue progression between ischemic and hemorrhagic strokes. Fatigue tends to ameliorate over time in hemorrhagic stroke cases, paralleling functional recovery, while it remains stable over time in ischemic stroke cases. Conclusion: Our results underscore the detrimental impact of early poststroke fatigue on long-term outcomes. Furthermore, they highlight the imperative of managing poststroke fatigue, particularly during the subacute phase of stroke recovery.

Analysis of Differences in Running Demands between Official Matches and Transition Games of Young Professional Soccer Players according to the Playing Position.

The aim of this study was to compare the running demands of transition games (TGs) and official matches, analysing their requirements according to the performance of each position. An observational design was used to examine the activity of 20 soccer players during official matches and TGs. GPS technology was used to monitor the total distance covered (DC), distance at speeds between 14-17.9 km•h-1, 18-21 km•h-1, and above 21 km•h-1, peak speed, accelerations and decelerations above 2.5 m•s-2, and Player Load for both activities. All players were assigned to groups: centre-backs (CBs), fullbacks (FBs), defensive midfielders (DMFs), offensive midfielders (OMFs), wide midfielders (WMFs) and strikers (Ss). TGs showed greater total DC, DC 14-17.9 km•h-1, DC 18-21 km•h-1, DC >21 km•h-1, accelerations and decelerations >2.5 m•s-2, and Player Load (p < 0.01). CBs, FBs and Ss showed more DC, DC 14-17.9 km•h-1, DC 18-21 km•h-1, DC >21 km•h-1, accelerations and decelerations >2.5 m•s-2 and Player Load in TGs (p < 0.01). In the midfielder positions, transition game players showed greater DC 18-21 km•h-1, DC >21 km•h-1, accelerations and decelerations >2.5 m•s-2 than in matches (p < 0.05). DMFs showed higher total DC (p < 0.05) and WMFs greater DC and DC 14-17.9 km•h-1 (p < 0.01) in these drills. During transition games CBs showed greater DC 14-17.9 km•h-1 than FBs, and greater DC than Ss (p < 0.05). FBs performed more decelerations >2.5 m•s-2 than DMFs and OMFs (p < 0.05). TGs produced a homogenized load in soccer players, independent of their position, which exceeded the external load of official matches.

Vitamin D opposes multilineage cell differentiation induced by Notch inhibition and BMP4 pathway activation in human colon organoids.

Understanding the mechanisms involved in colonic epithelial differentiation is key to unraveling the alterations causing inflammatory conditions and cancer. Organoid cultures provide an unique tool to address these questions but studies are scarce. We report a differentiation system toward enterocytes and goblet cells, the two major colonic epithelial cell lineages, using colon organoids generated from healthy tissue of colorectal cancer patients. Culture of these organoids in medium lacking stemness agents resulted in a modest ultrastructural differentiation phenotype with low-level expression of enterocyte (KLF4, KRT20, CA1, FABP2) and goblet cell (TFF2, TFF3, AGR2) lineage markers. BMP pathway activation through depletion of Noggin and addition of BMP4 resulted in enterocyte-biased differentiation. Contrarily, blockade of the Notch pathway using the γ-secretase inhibitor dibenzazepine (DBZ) favored goblet cell differentiation. Combination treatment with BMP4 and DBZ caused a balanced strong induction of both lineages. In contrast, colon tumor organoids responded poorly to BMP4 showing only weak signals of cell differentiation, and were unresponsive to DBZ. We also investigated the effects of 1α,25-dihydroxyvitamin D3 (calcitriol) on differentiation. Calcitriol attenuated the effects of BMP4 and DBZ on colon normal organoids, with reduced expression of differentiation genes and phenotype. Consistently, in normal organoids, calcitriol inhibited early signaling by BMP4 as assessed by reduction of the level of phospho-SMAD1/5/8. Our results show that BMP and Notch signaling play key roles in human colon stem cell differentiation to the enterocytic and goblet cell lineages and that calcitriol modulates these processes favoring stemness features.

Influence of the Number of Players on the Load of Soccer Players During Transition Games.

The aims of this study were to determine the effect of different compositions in transition games (TGs) on the load of soccer players and to evaluate their performance in physical tests. Using a GPS system, 18 players were monitored during: 3vs2, 2vs1 and 1vs1. Distance covered (DC), DC 18-20.9 km·h-1, 21-23.9 km·h-1,>24 km·h-1, peak speed, accelerations (Acc) and decelerations (Dec)>1.0 m·s-2 and>2.5 m·s-2 and rate of perceived exertion (RPE) were recorded. Before and after each TG, countermovement-jump (CMJ), 15- (S15) and 30 m (S30) speed tests were assessed. TG3vs2 showed greater DC and Dec>1.0 m·s-2 than TG2vs1, and DC, DC 18.0-23.9 km·h-1, Acc>1.0 m·s-2 and Dec>2.5 m·s-2 than TG1vs1 (p<0.01). TG2vs1 achieved higher DC, DC 18.0-23.9 km·h-1, and Acc>2.5 m·s-2 (p<0.01) but lower peak speed (p=0.02) and RPE (p=0.02) than TG1vs1. Post-intervention, TG1vs1 showed lower CMJ and higher S15 (p=0.02), while TG3vs2, showed improvements in CMJ (p<0.01). The three tasks showed large variations for DC>24 km·h-1, Acc>1.0 m·s-2, Dec>1.0 m·s-2 and Dec>2.5 m·s-2. The load of TGs is sensitive to their player composition.

Human Purkinje cells outperform mouse Purkinje cells in dendritic complexity and computational capacity.

Purkinje cells in the cerebellum are among the largest neurons in the brain and have been extensively investigated in rodents. However, their morphological and physiological properties remain poorly understood in humans. In this study, we utilized high-resolution morphological reconstructions and unique electrophysiological recordings of human Purkinje cells ex vivo to generate computational models and estimate computational capacity. An inter-species comparison showed that human Purkinje cell had similar fractal structures but were larger than those of mouse Purkinje cells. Consequently, given a similar spine density (2/µm), human Purkinje cell hosted approximately 7.5 times more dendritic spines than those of mice. Moreover, human Purkinje cells had a higher dendritic complexity than mouse Purkinje cells and usually emitted 2-3 main dendritic trunks instead of one. Intrinsic electro-responsiveness was similar between the two species, but model simulations revealed that the dendrites could process ~6.5 times (n = 51 vs. n = 8) more input patterns in human Purkinje cells than in mouse Purkinje cells. Thus, while human Purkinje cells maintained spike discharge properties similar to those of rodents during evolution, they developed more complex dendrites, enhancing computational capacity.

Relative Match Load in Young Professional Soccer Players during Soccer-7 and Soccer-11.

The aim of this study was to analyze the differences in internal and external load during Soccer-7 and Soccer-11, comparing positional requirements and neuromuscular fatigue in both modalities. Twenty-four young soccer players were monitored in Soccer-7 and Soccer-11 matches using global positioning systems. Total distance covered (TD), distance covered at high speed (HSR), distance covered at very high speed (VHSR), peak speed, accelerations (Acc) and decelerations (Dec) and rate of perceived exertion (RPE) were recorded differentiating between central backs (CB), midfielders (MF), external players (EX) and forwards (FW). Neuromuscular fatigue were assessed using a jump test. During Soccer-11, players showed significantly higher TD, HSR and VHSR, with low Acc and greater RPE compared with Soccer-7. During Soccer-11, all positions recorded significantly greater TD, distance at HSR and at VHSR than Soccer-7. In terms of playing position, CB, MF and FW achieved significantly higher Peak Speed during Soccer-1, but there was no difference for EX. During Soccer-7 all positions performed significantly higher numbers of Acc. Although the Soccer-7 modality is considered an optimal format for the development of young soccer players, there is a significant difference in match running activity for all playing positions with respect to the Soccer-11 format.

Observing the primary steps of ion solvation in helium droplets.

Solvation is a ubiquitous phenomenon in the natural sciences. At the macroscopic level, it is well understood through thermodynamics and chemical reaction kinetics1,2. At the atomic level, the primary steps of solvation are the attraction and binding of individual molecules or atoms of a solvent to molecules or ions of a solute1. These steps have, however, never been observed in real time. Here we instantly create a single sodium ion at the surface of a liquid helium nanodroplet3,4, and measure the number of solvent atoms that successively attach to the ion as a function of time. We found that the binding dynamics of the first five helium atoms is well described by a Poissonian process with a binding rate of 2.0 atoms per picosecond. This rate is consistent with time-dependent density-functional-theory simulations of the solvation process. Furthermore, our measurements enable an estimate of the energy removed from the region around the sodium ion as a function of time, revealing that half of the total solvation energy is dissipated after four picoseconds. Our experimental method opens possibilities for benchmarking theoretical models of ion solvation and for time-resolved measurements of cation-molecule complex formation.

Different pitch configurations constrain the external and internal loads of young professional soccer players during transition games.

The aims of this study were to compare the influence of transition game (TG) size on the external and internal loads of young professional soccer players and to describe the high-speed profile of these drills in response to pitch dimensions. Eighteen young professional soccer players (age: 16.1 ± 0.3 years; height: 178.3 ± 5.4 cm; weight: 70.1 ± 6.2 kg) performed a 3vs2 TG on pitches measuring 40 × 30 m (TG30), 40 × 50 m (TG50) and 40 × 70 m (TG70) m. Distance covered (DC); accelerations-decelerations above 1.0 m · s-2 and 2.5 m · s-2; rate of perceived exertion (RPE); maximal heart rate and time above 90%; DC at 18.0 to 21.0 km · h-1 (DC 18-20.9 km · h-1); DC at 21.0 to 23.9 km · h-1 (DC 21-23.9 km · h-1); DC above 24.0 km · h-1 (DC > 24 km · h-1); and peak speed and sprint profile (duration, distance and maximal speed) were measured. TG30 achieved lower DC, DC above 18 km · h-1, peak speed, sprint distance and RPE than TG50 and TG70 (p < 0.01 and p < 0.05) and lower sprint duration and maximal speed sprint than TG70 (p < 0.01). TG30 and TG50 achieved higher Acc > 1.0 and > 2.5 m · s-2 respectively than TG70 (p < 0.05). TG70 showed greater DC above 21 km · h-1, peak speed, sprint distance and maximal speed sprint than TG50 (p < 0.01). Soccer coaches should use larger TGs to overload variables related to high speed and sprint demands during training and smaller TG formats to stimulate the accelerations of the soccer players.

Direct conversion of amino acids to oxetanol bioisosteres via photoredox catalysis.

Carboxylic acids are an important structural feature in many drugs, but are associated with a number of unfavorable pharmacological properties. To address this problem, carboxylic acids can be replaced with bioisosteric mimics that interact similarly with biological targets but avoid these liabilities. Recently, 3-oxetanols have been identified as useful carboxylic acid bioisosteres that maintain similar hydrogen-bonding capacity while decreasing acidity and increasing lipophilicity. However, the installation of 3-oxetanols generally requires multistep de novo synthesis, presenting an obstacle to investigation of these promising bioisosteres. Herein, we report a new synthetic approach involving direct conversion of carboxylic acids to 3-oxetanols using a photoredox-catalyzed decarboxylative addition to 3-oxetanone. Two versions of the transformation have been developed, in the presence or absence of CrCl3 and TMSCl cocatalysts. The reactions are effective for a variety of N-aryl α-amino acids and have excellent functional group tolerance. The Cr-free conditions generally provide higher yields and avoid the use of chromium reagents. Further, the Cr-free conditions were extended to a series of N,N-dialkyl α-amino acid substrates. Mechanistic studies suggest that the Cr-mediated reaction proceeds predominantly via in situ formation of an alkyl-Cr intermediate while the Cr-free reaction proceeds largely via radical addition to a Brønsted acid-activated ketone. Chain propagation processes provide quantum yields of 5 and 10, respectively.

Analysis of Connectome Graphs Based on Boundary Scale.

The purpose of this work is to advance in the computational study of connectome graphs from a topological point of view. Specifically, starting from a sequence of hypergraphs associated to a brain graph (obtained using the Boundary Scale model, BS2), we analyze the resulting scale-space representation using classical topological features, such as Betti numbers and average node and edge degrees. In this way, the topological information that can be extracted from the original graph is substantially enriched, thus providing an insightful description of the graph from a clinical perspective. To assess the qualitative and quantitative topological information gain of the BS2 model, we carried out an empirical analysis of neuroimaging data using a dataset that contains the connectomes of 96 healthy subjects, 52 women and 44 men, generated from MRI scans in the Human Connectome Project. The results obtained shed light on the differences between these two classes of subjects in terms of neural connectivity.

Clinico-Pathological Features, Outcomes and Impacts of COVID-19 Pandemic on Patients with Early-Onset Colorectal Cancer: A Single-Institution Experience.

BACKGROUND: The rising incidence of colorectal cancer (CRC) among young patients is alarming. We aim to characterize the clinico-pathological features and outcomes of patients with early-onset CRC (EOCRC), as well as the impacts of COVID-19 pandemic. METHODS: We included all patients with pathologically confirmed diagnoses of CRC at Hospital Universitario La Paz from October 2016 to December 2021. The EOCRC cut-off age was 50 years old. RESULTS: A total of 1475 patients diagnosed with CRC were included, eighty (5.4%) of whom had EOCRC. Significant differences were found between EOCRC and later-onset patients regarding T, N stage and metastatic presentation at diagnosis; perineural invasion; tumor budding; high-grade tumors; and signet ring cell histology, with all issues having higher prevalence in the early-onset group. More EOCRC patients had the RAS/ BRAF wild type. Chemotherapy was administered more frequently to patients with EOCRC. In the metastatic setting, the EOCRC group presented a significantly longer median OS. Regarding the COVID-19 pandemic, more patients with COVID-19 were diagnosed with metastatic disease (61%) in the year after the lockdown (14 March 2020) than in the pre-pandemic EOCRC group (29%). CONCLUSIONS: EOCRC is diagnosed at a more advanced stage and with worse survival features in localized patients. More patients with EOCRC were diagnosed with metastatic disease in the year after the COVID-19 pandemic lockdown. The long-term consequences of COVID-19 are yet to be determined.

Vitamin D induces SIRT1 activation through K610 deacetylation in colon cancer.

Posttranslational modifications of epigenetic modifiers provide a flexible and timely mechanism for rapid adaptations to the dynamic environment of cancer cells. SIRT1 is an NAD+-dependent epigenetic modifier whose activity is classically associated with healthy aging and longevity, but its function in cancer is not well understood. Here, we reveal that 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3, calcitriol), the active metabolite of vitamin D (VD), promotes SIRT1 activation through auto-deacetylation in human colon carcinoma cells, and identify lysine 610 as an essential driver of SIRT1 activity. Remarkably, our data show that the post-translational control of SIRT1 activity mediates the antiproliferative action of 1,25(OH)2D3. This effect is reproduced by the SIRT1 activator SRT1720, suggesting that SIRT1 activators may offer new therapeutic possibilities for colon cancer patients who are VD deficient or unresponsive. Moreover, this might be extrapolated to inflammation and other VD deficiency-associated and highly prevalent diseases in which SIRT1 plays a prominent role.

An amino acid transporter subunit as an antibody-drug conjugate target in colorectal cancer.

BACKGROUND: Advanced colorectal cancer (CRC) is difficult to treat. For that reason, the development of novel therapeutics is necessary. Here we describe a potentially actionable plasma membrane target, the amino acid transporter protein subunit CD98hc. METHODS: Western blot and immunohistochemical analyses of CD98hc protein expression were carried out on paired normal and tumoral tissues from patients with CRC. Immunofluorescence and western studies were used to characterize the action of a DM1-based CD98hc-directed antibody-drug conjugate (ADC). MTT and Annexin V studies were performed to evaluate the effect of the anti-CD98hc-ADC on cell proliferation and apoptosis. CRISPR/Cas9 and shRNA were used to explore the specificity of the ADC. In vitro analyses of the antitumoral activity of the anti-CD98hc-ADC on 3D patient-derived normal as well as tumoral organoids were also carried out. Xenografted CRC cells and a PDX were used to analyze the antitumoral properties of the anti-CD98hc-ADC. RESULTS: Genomic as well proteomic analyses of paired normal and tumoral samples showed that CD98hc expression was significantly higher in tumoral tissues as compared to levels of CD98hc present in the normal colonic tissue. In human CRC cell lines, an ADC that recognized the CD98hc ectodomain, reached the lysosomes and exerted potent antitumoral activity. The specificity of the CD98hc-directed ADC was demonstrated using CRC cells in which CD98hc was decreased by shRNA or deleted using CRISPR/Cas9. Studies in patient-derived organoids verified the antitumoral action of the anti-CD98hc-ADC, which largely spared normal tissue-derived colon organoids. In vivo studies using xenografted CRC cells or patient-derived xenografts confirmed the antitumoral activity of the anti-CD98hc-ADC. CONCLUSIONS: The studies herewith reported indicate that CD98hc may represent a novel ADC target that, upon well-designed clinical trials, could be used to increase the therapeutic armamentarium against CRC.