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Objective: To describe the epidemiology of quadriceps muscle strain injury (QMSI) in elite Australian Football League (AFL) players, explore recovery milestones and determine whether recovery is impacted by factors such as injury type (index vs. re-injury), the primary muscle injured and the mechanism of injury., Measures: All QMSI data reported to the Soft Tissue Injury Registry of the AFL from the 2014 to 2020 seasons were evaluated. Player demographic data, circumstances of injury, MRI reports and recovery outcomes following injury were extracted. Descriptive statistics and frequency distributions are presented. Recovery outcomes for injury type, primary muscle injured and the mechanism of injury were compared using univariate analyses., Results: There were 164 QMSIs from 122 players reported (134 index; 30 re-injuries). Almost all (91.3%) QMSIs involved the rectus femoris. Half (48.4%) of the QMSIs occurred during kicking and most commonly affected the dominant kicking leg (72%). The majority occurred at training (64.6%). All re-injuries involved the rectus femoris, most occurred from kicking (63.0%) and within 6 months of the preceding injury (70%). The mean return to play (RTP) time was 25.4 days (95%CI = 22.6-28.2) and rectus femoris injuries took around 14 days longer to RTP than vastii injuries (p = 0.001). QMSIs with a kicking mechanism took the longest to RTP of all injury mechanisms., Conclusion: In AFL players, QMSIs occur mostly in the dominant leg from a kicking mechanism. Rectus femoris injuries are more prevalent and result in longer RTP time frames. Re-injuries exclusively involved the rectus femoris, primarily from kicking., (© 2023 The Authors. Scandinavian Journal of Medicine & Science In Sports published by John Wiley & Sons Ltd.)

Plant diversity can increase productivity. One mechanism behind this biodiversity effect is facilitation, which is when one species increases the performance of another species. Plants with extrafloral nectaries (EFNs) establish defense mutualisms with ants. However, whether EFN plants facilitate defense of neighboring non-EFN plants is unknown. Synthesizing data on ants, herbivores, leaf damage, and defense traits from a forest biodiversity experiment, we show that trees growing adjacent to EFN trees had higher ant biomass and species richness and lower caterpillar biomass than conspecific controls without EFN-bearing neighbors. Concurrently, the composition of defense traits in non-EFN trees changed. Thus, when non-EFN trees benefit from lower herbivore loads as a result of ants spilling over from EFN tree neighbors, this may allow relatively reduced resource allocation to defense in the former, potentially explaining the higher growth of those trees. Via this mutualist-mediated facilitation, promoting EFN trees in tropical reforestation could foster carbon capture and multiple other ecosystem functions., (© 2023 The Ecological Society of America.)

Purpose: To evaluate the safety and efficacy of matrix-associated autologous chondrocyte implantation (ACI) using spheroids in comparison to arthroscopic microfracture for the treatment of symptomatic cartilage defects of the knee., Methods: In a prospective multicenter-controlled trial, patients aged between 18 and 50 years, with single symptomatic focal cartilage defects between 1 and 4 cm 2 (mean 2.6 ± 0.8, median 2.75, range 1.44-5.00) in the knee were randomized to treatment with ACI with spheroids (n = 52) or microfracture (n = 50). Primary clinical outcome was assessed by the Knee Injury and Osteoarthritis Outcome Score (KOOS). Analyses were performed in a defined hierarchical manner where outcomes of ACI were first compared to baseline values followed by a comparison to the microfracture group with repeated-measures ANCOVA with a non-inferiority approach. Subgroup analyses were performed to investigate the influence of age and defect size on the overall KOOS. Secondary clinical outcomes were the magnetic resonance observation of cartilage repair tissue (MOCART), modified Lysholm score and International Knee Documentation Committee (IKDC) examination form. Safety data focused on adverse events. Here the 5 years results are presented at which there were 33 observed cases in the ACI group and 30 in the microfracture group., Results: The overall KOOS and its five subscores were significantly improved compared to baseline for both the ACI and microfracture group. Non-inferiority of ACI to microfracture was confirmed for the overall KOOS and the subscores, while for the subscores activities of daily living, quality of life and sports and recreation of the threshold for superiority was passed. In the ACI group, a notably more rapid initial improvement of the KOOS was found at three months for the older age group compared to the younger age group and the microfracture group. No other differences were found based on age or defect size. In addition, clinical improvement was found for the MOCART, modified Lysholm and IKDC examination form both the ACI and microfracture group. No safety concern related to either treatment was observed., Conclusion: This study confirms the safety and efficacy of matrix-associated ACI with spheroids at a mid to long-term follow-up. Non-inferiority of ACI to microfracture was confirmed for the overall KOOS and all subscores, while superiority was reached for the subscores activities of daily living, quality of life and sports and recreation in the ACI group. This underlines the importance of ACI for the young and active patients., Level of Evidence: I., (© 2022. The Author(s) under exclusive licence to European Society of Sports Traumatology, Knee Surgery, Arthroscopy (ESSKA).)

Nitric oxide (NO) is a signaling molecule produced by NO synthases (NOS1-3) to control processes such as neurotransmission, vascular permeability, and immune function. Although myeloid cell-derived NO has been shown to suppress T-cell responses, the role of NO synthesis in T cells themselves is not well understood. Here, we showed that significant amounts of NO were synthesized in human and murine CD8+ T cells following activation. Tumor growth was significantly accelerated in a T cell-specific, Nos2-null mouse model. Genetic deletion of Nos2 expression in murine T cells altered effector differentiation, reduced tumor infiltration, and inhibited recall responses and adoptive cell transfer function. These data show that endogenous NO production plays a critical role in T cell-mediated tumor immunity., (©2022 The Authors; Published by the American Association for Cancer Research.)

High tibial osteotomy (HTO) represents a sensible treatment option for patients with moderate unicondylar osteoarthritis of the knee and extraarticular malalignment. The possibility of a continuously variable correction setting and a surgical approach low in complications has meant that the medial opening osteotomy has prevailed over the past decades. The objective of the present study was to determine whether anteromedially positioned small plates are nevertheless forgiving under biomechanically unfavourable conditions (overcorrection and lateral hinge fracture). In this study, a simulated HTO was performed on composite tibiae with a 10-mm wedge and fixed-angle anteromedial osteosynthesis with a small implant. Force was applied axially in a neutral mechanical axis, a slight and a marked overcorrection into valgus, with and without a lateral hinge fracture in each case. At the same time, a physiological gait with a dual-peak force profile and a peak load of 2.4 kN was simulated. Interfragmentary motion and rigidity were determined. The rigidity of the osteosynthesis increased over the cycles investigated. A slight overcorrection into valgus led to the lowest interfragmentary motion, compared with pronounced valgisation and neutral alignment. A lateral hinge fracture led to a significant decrease in rigidity and increase in interfragmentary motion. However, in no case was the limit of 1 mm interfragmentary motion critical for osteotomy healing exceeded. The degree of correction of the leg axis, and the presence of a lateral hinge fracture, have an influence on rigidity and interfragmentary motion. From a mechanically neutral axis ranging up to pronounced overcorrection, the implant investigated offers sufficient stability to allow healing of the osteotomy, even if a lateral hinge fracture is present.

Background: Many different sedation concepts for magnetic resonance imaging have been described for prematurely and term-born infants, ranging from "no sedation" to general anesthesia. Dexmedetomidine is an alpha-2 receptor agonist that is frequently used to sedate older children, because the anesthesiologist can easily adjust sedation depth, the patient maintains spontaneous breathing, and awakens rapidly afterwards., Aims: The present study evaluates whether dexmedetomidine could safely be used as the sole sedative for prematurely and term-born infants less than 60 weeks postconceptional age undergoing diagnostic procedures., Methods: We performed a retrospective monocentric analysis of n = 39 prematurely and term-born infants (<60 weeks postconceptional age or a body weight <5 kg) who were sedated with dexmedetomidine for an MRI at a German university hospital from August 2016 to November 2018., Results: Successful imaging was achieved in all cases. The median initial bolus of dexmedetomidine administered over 10 min was 1.39 μg kg -1 body weight (range 0.34-3.64 μg kg -1 ), followed with a continuous infusion at a median rate of 1.00 μg kg -1  h -1 (range 0.5-3.5 μg kg -1  h -1 ); however, 3 patients (7%) needed some additional sedation (ketamine or propofol). All patients, including 10 infants who had previously required respiratory support, underwent the procedure without any relevant desaturation or apnea. Bradycardia was observed in up to 15 out of 39 cases (38.5%), but only four (10.3% in total and 26.7% of bradycardia) required atropine., Conclusions: These results indicate that dexmedetomidine can be safely used for procedural sedation in the high-risk cohort of prematurely and term-born infants less than 60 weeks postconceptional age. Apnea during procedural sedation and subsequent stay in the recovery room is avoided, but bradycardia remains a relevant risk that may require treatment., (© 2022 The Authors. Pediatric Anesthesia published by John Wiley & Sons Ltd.)

Purpose of Review: The purpose of this overview is to analyze existing reviews on psychosocial interventions for children of parents with cancer and synthesize implications for further practice, interventions, and research in this field. The aim of this overview is therefore to generate systematic knowledge about what can be classed as evidence-based knowledge in this field., Recent Findings: The literature search in MEDLINE, CINAHL, PsycINFO, PSYNDEX, and PsycARTICLES resulted in three systematic reviews, which were evaluated by the AMSTAR-2-tool for quality assessment and the PRISMA-checklist for reporting. Results were analyzed through narrative synthesis due to the heterogeneity of the studies. The three systematic reviews were evaluated by the AMSTAR-2-tool for quality assessment and the PRISMA-checklist for reporting. AMSTAR-2 revealed critically low quality for all three reviews but taking into account the study situation of this scientific context, a more optimistic quality assessment can be suggested. The PRISMA checklist revealed good results. Positive evidence was found for the effect of psychosocial interventions concerning depressive symptoms, children's behavior, communication within the family, and quality of life. A comparison of the interventions is not possible due to the high degree of heterogeneity of the studies., Summary: Following the principles of evidence-based medicine, this overview, together with clinical-practical expertise and the needs of those affected, could contribute to evidence-based care and stimulate future guidelines in this important field. The valuable engagement with questions around evidence-based practice invites professionals and researchers to enter into a common discourse to ultimately contribute to an improvement of the life situation of children of parents with cancer., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)

New borylation methodologies have been reported recently, wherein diboron(4) compounds apparently participate in free radical couplings via the homolytic cleavage of the B-B bond. We report herein that bis-NHC adducts of the type (NHC) 2 ·B 2 (OR) 4 , which are thermally unstable and undergo intramolecular ring expansion reactions (RER), are sources of boryl radicals of the type NHC-BR 2 ˙, exemplified by Me 2 Im Me ·Bneop˙ 1a (Me 2 Im Me = 1,3,4,5-tetramethyl-imidazolin-2-ylidene, neop = neopentylglycolato), which are formed by homolytic B-B bond cleavage. Attempts to apply the boryl moiety 1a in a metal-free borylation reaction by suppressing the RER failed. However, based on these findings, a protocol was developed using Me 2 Im Me ·B 2 pin 2 3 for the transition metal- and additive-free boryl transfer to substituted aryl iodides and bromides giving aryl boronate esters in good yields. Analysis of the side products and further studies concerning the reaction mechanism revealed that radicals are likely involved. An aryl radical was trapped by TEMPO, an EPR resonance, which was suggestive of a boron-based radical, was detected in situ , and running the reaction in styrene led to the formation of polystyrene. The isolation of a boronium cation side product, [(Me 2 Im Me ) 2 ·Bpin] + I - 7, demonstrated the fate of the second boryl moiety of B 2 pin 2 . Interestingly, Me 2 Im Me NHC reacts with aryl iodides and bromides generating radicals. A mechanism for the boryl radical transfer from Me 2 Im Me ·B 2 pin 2 3 to aryl iodides and bromides is proposed based on these experimental observations., Competing Interests: There are no conflicts of interest to declare., (This journal is © The Royal Society of Chemistry.)

Objective: To identify risk factors for quadriceps muscle strain injury in sport., Design: Risk factor systematic review., Literature Search: A systematic search was conducted in the MEDLINECINAHL, Embase, AMED, AUSPORT, SPORTDiscus, PEDro, and Cochrane Library databases (from inception to September 2021)., Study Selection Criteria: Studies reporting prospective data to evaluate risk factors related to index and/or recurrent quadriceps muscle strain injury., Data Synthesis: A risk-of-bias assessment (using a modified Quality in Prognosis Studies tool) was performed, and we used best-evidence synthesis to qualitatively synthesize the data to quantify relationships between risk factors and quadriceps muscle injury., Results: Sixteen studies were included, capturing 2408 quadriceps injuries in 11 719 athletes. Meta-analyses were not performed due to clinical heterogeneity. The dominant kicking leg (over 3154 individuals, 1055 injuries), a previous history of quadriceps muscle injury (6208 individuals, 975 injuries), and a recent history of hamstring strain (4087 individuals, 581 injuries) were intrinsic factors associated with quadriceps injury. Extrinsic factors relating to the preseason period and competitive match play increased quadriceps injury risk; participating at higher levels of competition decreased quadriceps injury risk. Age, weight, and flexibility (intrinsic factors) had no association with quadriceps injury., Conclusion: Previous quadriceps injury, recent hamstring injury, the dominant kicking leg, and competitive match play were the strongest risk factors for future quadriceps muscle injury in sport. J Orthop Sports Phys Ther 2022;52(6):389-400. doi:10.2519/jospt.2022.10870 .

Objective: Matrix-associated autologous chondrocyte implantation (ACI) and microfracture (MF) are well-established treatments for cartilage defects of the knee. However, high-level evidence comparing microfracture and spheroid technology ACI is limited., Design: Prospective, phase III clinical trial with patients randomized to ACI ( N = 52) or MF ( N = 50). Level of evidence: 1, randomized controlled trial. Both procedures followed standard protocols. For ACI 10 to 70 spheroids/cm 2 were administered. Primary outcome measure was the Knee Injury and Osteoarthritis Outcome Score (KOOS). This report presents results for 36 months after treatment., Results: Both ACI and MF showed significant improvement over the entire 3-year observation period. For the overall KOOS, noninferiority of ACI (the intended primary goal of the study) was formally confirmed; additionally, for the subscores "Activities of Daily Living" and "Sport and Recreation," superiority of ACI over MF was shown at descriptive level. Occurrence of adverse events were not different between both treatments (ACI 77%; MF 74%). Four patients in the MF group required reoperation which was defined as treatment failure. No treatment failure was reported for the ACI group., Conclusions: Patients treated with matrix-associated ACI with spheroid technology showed substantial improvement in various clinical outcomes after 36 months. The advantages of ACI compared with microfracture was underlined by demonstrating noninferiority, in overall KOOS and superiority in the KOOS subscores "Activities of Daily Living" and "Sport and Recreation." In the present study, subgroups comparing different age groups and defect sizes showed comparable clinical outcomes.

Objective: To determine the reliability of range-of-motion (ROM) measurements and describe physiologic differences in ROM or habituation effects during gait analysis of healthy dogs walking on a treadmill., Animals: 11 orthopedically normal dogs., Procedures: ROM of appendicular joints was determined for each dog while walking on a treadmill on 3 consecutive examination days and once again 6 weeks later. Significant differences in ROM between examination days were determined and quantified. As a measure of reproducibility, the coefficient of variation for repeated measurements was calculated, as were the minimum differences necessary to distinguish between physiologic variation and true change in ROM., Results: Mean ROM of the shoulder, elbow, and carpal joints varied among examination days between 29.9° and 33.1°, 49.4° and 52.8°, and 7.7° and 88.1°, respectively. Mean associated minimum differences were 12.0°, 14.1°, and 35.6°. Mean ROM of the hip, knee, and tarsal joints varied between 32.9° and 35.8°, 33.7° and 36.8°, and 31.7° and 33.5°, respectively. Mean associated minimum differences were 16.2°, 14.0°, and 9.2°. Only ROM of the elbow joint was reproducible to a small degree. Few systematic effects were detected., Conclusions and Clinical Relevance: Measurement of ROM in healthy dogs walking on a treadmill was shown to be diagnostically unreliable owing to high variation among examination days. However, random physiologic fluctuations could be distinguished from systematic effects, demonstrating the importance of reliably applicable threshold values for follow-up treadmill examinations. The applicability of the minimum differences determined here to orthopedically diseased dogs remains to be determined.

Axonal degeneration (AxD) is a pathological hallmark of many neurodegenerative diseases. Deciphering the morphological patterns of AxD will help to understand the underlying mechanisms and develop effective therapies. Here, we evaluated the progression of AxD in cortical neurons using a novel microfluidic device together with a deep learning tool that we developed for the enhanced-throughput analysis of AxD on microscopic images. The trained convolutional neural network (CNN) sensitively and specifically segmented the features of AxD including axons, axonal swellings, and axonal fragments. Its performance exceeded that of the human evaluators. In an in vitro model of AxD in hemorrhagic stroke induced by the hemolysis product hemin, we detected a time-dependent degeneration of axons leading to a decrease in axon area, while axonal swelling and fragment areas increased. Axonal swellings preceded axon fragmentation, suggesting that swellings may be reliable predictors of AxD. Using a recurrent neural network (RNN), we identified four morphological patterns of AxD (granular, retraction, swelling, and transport degeneration). These findings indicate a morphological heterogeneity of AxD in hemorrhagic stroke. Our EntireAxon platform enables the systematic analysis of axons and AxD in time-lapse microscopy and unravels a so-far unknown intricacy in which AxD can occur in a disease context.

Preterm infants frequently suffer from pulmonary complications due to a physiological and structural lung immaturity resulting in significant morbidity and mortality. Novel in vitro and in vivo models are required to study the underlying mechanisms of late lung maturation and to facilitate the development of new therapeutic strategies. Organoids recapitulate essential aspects of structural organization and possibly organ function, and can be used to model developmental and disease processes. We aimed at generating fetal lung organoids (LOs) and to functionally characterize this in vitro model in comparison to primary lung epithelial cells and lung explants ex vivo . LOs were generated with alveolar and endothelial cells from fetal rat lung tissue, using a Matrigel-gradient and air-liquid-interface culture conditions. Immunocytochemical analysis showed that the LOs consisted of polarized epithelial cell adhesion molecule (EpCAM)-positive cells with the apical membrane compartment facing the organoid lumen. Expression of the alveolar type 2 cell marker, RT2-70, and the Club cell marker, CC-10, were observed. Na + transporter and surfactant protein mRNA expression were detected in the LOs. First time patch clamp analyses demonstrated the presence of several ion channels with specific electrophysiological properties, comparable to vital lung slices. Furthermore, the responsiveness of LOs to glucocorticoids was demonstrated. Finally, maturation of LOs induced by mesenchymal stem cells confirmed the convenience of the model to test and establish novel therapeutic strategies. The results showed that fetal LOs replicate key biological lung functions essential for lung maturation and therefore constitute a suitable in vitro model system to study lung development and related diseases., 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 Laube, Pietsch, Pannicke, Thome and Fabian.)

Global vegetation and land-surface models embody interdisciplinary scientific understanding of the behaviour of plants and ecosystems, and are indispensable to project the impacts of environmental change on vegetation and the interactions between vegetation and climate. However, systematic errors and persistently large differences among carbon and water cycle projections by different models highlight the limitations of current process formulations. In this review, focusing on core plant functions in the terrestrial carbon and water cycles, we show how unifying hypotheses derived from eco-evolutionary optimality (EEO) principles can provide novel, parameter-sparse representations of plant and vegetation processes. We present case studies that demonstrate how EEO generates parsimonious representations of core, leaf-level processes that are individually testable and supported by evidence. EEO approaches to photosynthesis and primary production, dark respiration and stomatal behaviour are ripe for implementation in global models. EEO approaches to other important traits, including the leaf economics spectrum and applications of EEO at the community level are active research areas. Independently tested modules emerging from EEO studies could profitably be integrated into modelling frameworks that account for the multiple time scales on which plants and plant communities adjust to environmental change., (© 2021 The Authors New Phytologist © 2021 New Phytologist Foundation.)

Planarians are able to stand long periods of starvation by maintaining adult stem cell pools and regenerative capacity. The molecular pathways that are needed for the maintenance of regeneration during starvation are not known. Here, we show that down-regulation of chaperonin TRiC/CCT subunits abrogates the regeneration capacity of planarians during starvation, but TRiC/CCT subunits are dispensable for regeneration in fed planarians. Under starvation, they are required to maintain mitotic fidelity and for blastema formation. We show that TRiC subunits modulate the unfolded protein response (UPR) and are required to maintain ATP levels in starved planarians. Regenerative defects in starved CCT-depleted planarians can be rescued by either chemical induction of mild endoplasmic reticulum stress, which leads to induction of the UPR, or by the supplementation of fatty acids. Together, these results indicate that CCT-dependent UPR induction promotes regeneration of planarians under food restriction., (© 2021 The Authors. Published under the terms of the CC BY 4.0 license.)

Background: Nowadays, osteotomy near the knee joint for axial deformities has become an indispensable surgical procedure for joint preservation in the orthopaedic-surgical spectrum. The exact localization and analysis of the deformity are crucial to restoring physiological loading conditions of the entire leg by means of a suitable osteotomy., Procedures: Thus, above all, the medial "open-wedge" osteotomy has established itself as a standard procedure for the treatment of gonarthrosis caused by varus malalignment. Furthermore, the varus closed-wedge osteotomy of the distal femur also shows very good long-term results. Basically, osteotomies close to the knee are causal corrections in which biological and mechanical aspects must be taken into account. Thus, the correct indication, attention to risk factors, and surgical technique determine the long-term success of the procedure. The doctrine of femoral valgus deformity and the resulting lateral arthrosis, on the other hand, must be reconsidered according to the latest knowledge.

Lipid metabolism influences stem cell maintenance and differentiation but genetic factors that control these processes remain to be delineated. Here, we identify Tnfaip2 as an inhibitor of reprogramming of mouse fibroblasts into induced pluripotent stem cells. Tnfaip2 knockout impairs differentiation of embryonic stem cells (ESCs), and knockdown of the planarian para-ortholog, Smed-exoc3, abrogates in vivo tissue homeostasis and regeneration-processes that are driven by somatic stem cells. When stimulated to differentiate, Tnfaip2-deficient ESCs fail to induce synthesis of cellular triacylglycerol (TAG) and lipid droplets (LD) coinciding with reduced expression of vimentin (Vim)-a known inducer of LD formation. Smed-exoc3 depletion also causes a strong reduction of TAGs in planarians. The study shows that Tnfaip2 acts epistatically with and upstream of Vim in impairing cellular reprogramming. Supplementing palmitic acid (PA) and palmitoyl-L-carnitine (the mobilized form of PA) restores the differentiation capacity of Tnfaip2-deficient ESCs and organ maintenance in Smed-exoc3-depleted planarians. Together, these results identify a novel role of Tnfaip2 and exoc3 in controlling lipid metabolism, which is essential for ESC differentiation and planarian organ maintenance., (© 2020 The Authors Published under the terms of the CC BY 4.0 license.)

Objective: To investigate whether an actual improvement in gait could be differentiated from physiologic differences or habituation effects during gait analysis of dogs., Animals: 11 healthy dogs., Procedures: On 4 examination days, kinetic parameters were measured while dogs were walking on a treadmill. Differences in mean parameter values and habituation effects (ie, effect sizes) were quantified and compared among examination days. Coefficients of variation for repeated measurements were calculated to determine measurement reproducibility, and minimum differences were calculated to distinguish between physiologic fluctuation and an actual change in gait pattern., Results: Among the 4 examination days, mean absolute differences in peak vertical force and vertical impulse (VI) varied from 1.5% to 5.3% of body weight (BW) and 0.9% to 1.8% of BW·s, respectively. Mean absolute differences in the percentage of stance-phase duration (%SPD) and relative stride length (RSL) varied from 0.9% to 3.2% and 1.7% to 3.0%, respectively. Reproducibility of parameter measurements was good. Values for %SPD had the lowest amount of dispersion and largest effect size, suggesting a habituation effect for this parameter. Calculated minimum differences among the days for peak vertical force, VI, %SPD, and RSL did not exceed 9.9% of BW, 3.3% of BW·s, 5.8 percentage points, and 5.2 percentage points, respectively., Conclusions and Clinical Relevance: The %SPD of healthy dogs walking on a treadmill was the most sensitive and diagnostically reliable of the measured kinetic parameters, in contrast to VI and RSL. Findings suggested that actual changes can be distinguished from random physiologic fluctuations during gait analysis of dogs.

Cancer immunotherapy is advancing rapidly and gene-modified T cells expressing chimeric antigen receptors (CARs) show particular promise. A challenge of CAR-T cell therapy is that the ex vivo-generated CAR-T cells become exhausted during expansion in culture, and do not persist when transferred back to patients. It has become clear that naive and memory CD8 T cells perform better than the total CD8 T-cell populations in CAR-T immunotherapy because of better expansion, antitumor activity, and persistence, which are necessary features for therapeutic success and prevention of disease relapse. However, memory CAR-T cells are rarely used in the clinic due to generation challenges. We previously reported that mouse CD8 T cells cultured with the S enantiomer of the immunometabolite 2-hydroxyglutarate (S-2HG) exhibit enhanced antitumor activity. Here, we show that clinical-grade human donor CAR-T cells can be generated from naive precursors after culture with S-2HG. S-2HG-treated CAR-T cells establish long-term memory cells in vivo and show superior antitumor responses when compared with CAR-T cells generated with standard clinical protocols. This study provides the basis for a phase 1 clinical trial evaluating the activity of S-2HG-treated CD19-CAR-T cells in patients with B-cell malignancies., (© 2020 by The American Society of Hematology.)

Background: While effective interventions have been developed to support families where a parent has a mental illness in Adult Mental Health Services, embedding and sustaining them is challenging resulting in families not having access to support. This study developed an explanatory model of influencers that had enabled sustainability of the Let's Talk intervention in one service., Methods: A participatory case study was used to build an explanatory model of sustainability at the service using theoretical frameworks. Qualitative and quantitative data was collected about practitioner's practice and the organisation's implementation process and capacity to support practice. A local research group worked with the researcher using a transforming data approach through description, analysis and interpretation., Results: Influencers were grouped into four major categories: (1) External social, political and financial context, (2) Resources, (3) Prior organisational capacity and (4) Sustainability Factors. The last category, Sustainability factors, was divided into three subcategories: (4.1) Practitioner ( 4.2) Organisation and ( 4.3) Parent - Client . These categories form part of an explanatory model for the key influencers of continued practitioner practice and organisational capacity to support practice., Conclusions and Implications for Practice: In this case study, the pre-existing organisational context along with practitioner, organisation and parent-client factors operated together to influence sustainability. The results suggest that sustainability is more likely to be supported by both linking Let's Talk to existing organisational identity, capacity, structures and relationships and by supporting mutual adaptations to improve the fit. Additionally, by understanding that setbacks are common and ongoing adjustments are needed, implementers are able to have realistic expectations of sustainability., Competing Interests: Competing interestsThe authors declare they have no conflict of interest., (© The Author(s) 2020.)

Fluid-solid systems play a major role in a wide variety of industries, from pharmaceutical and consumer goods to chemical plants and energy generation. Along with this variety of fields comes a diversity in apparatuses and applications, most prominently fluidized and spouted beds, granulators and mixers, pneumatic conveying, drying, agglomeration, coating, and combustion. The most promising approach for modeling the flow in these systems is the CFD-DEM method, coupling computational fluid dynamics (CFD) for the fluid phase and the discrete element method (DEM) for the particles. This article reviews the progress in modeling particle-fluid flows with the CFD-DEM method. A brief overview of the basic method as well as methodical extensions of it are given. Recent applications of this simulation approach to separation and classification units, fluidized beds for both particle formation and energy conversion, comminution units, filtration, and bioreactors are reviewed. Future trends are identified and discussed regarding their viability.

Introduction: This study aimed to 1) identify the impact of external load variables on changes in wellness and 2) identify the impact of age, training/playing history, strength levels, and preseason loads on changes in wellness in elite Australian footballers., Methods: Data were collected from one team (45 athletes) during the 2017 season. Self-reported wellness was collected daily (4, best score possible; 28, worst score possible). External load/session availability variables were calculated using global positioning systems and session availability data from every training session and match. Additional variables included demographic data, preseason external loads, and strength/power measures. Linear mixed models were built and compared using root mean square error (RMSE) to determine the impact of variables on wellness., Results: The external load variables explained wellness to a large degree (RMSE = 1.55, 95% confidence intervals = 1.52 to 1.57). Modeling athlete ID as a random effect appeared to have the largest impact on wellness, improving the RMSE by 1.06 points. Aside from athlete ID, the variable that had the largest (albeit negligible) impact on wellness was sprint distance covered across preseason. Every additional 2.1 km covered across preseason worsened athletes' in-season wellness scores by 1.2 points (95% confidence intervals = 0.0-2.3)., Conclusions: The isolated impact of the individual variables on wellness was negligible. However, after accounting for the individual athlete variability, the external load variables examined collectively were able to explain wellness to a large extent. These results validate the sensitivity of wellness to monitor individual athletes' responses to the external loads imposed on them.

Plants and vegetation play a critical-but largely unpredictable-role in global environmental changes due to the multitude of contributing processes at widely different spatial and temporal scales. In this Perspective, we explore approaches to master this complexity and improve our ability to predict vegetation dynamics by explicitly taking account of principles that constrain plant and ecosystem behaviour: natural selection, self-organization and entropy maximization. These ideas are increasingly being used in vegetation models, but we argue that their full potential has yet to be realized. We demonstrate the power of natural selection-based optimality principles to predict photosynthetic and carbon allocation responses to multiple environmental drivers, as well as how individual plasticity leads to the predictable self-organization of forest canopies. We show how models of natural selection acting on a few key traits can generate realistic plant communities and how entropy maximization can identify the most probable outcomes of community dynamics in space- and time-varying environments. Finally, we present a roadmap indicating how these principles could be combined in a new generation of models with stronger theoretical foundations and an improved capacity to predict complex vegetation responses to environmental change.

Gut integrity impairment leading to increased intestinal permeability (IP) is hypothesized to be a trigger of critically illness. Approximately 15-20% of human ischemic stroke (IS) victims require intensive care, including patients with impaired level of consciousness or a high risk for developing life-threatening cerebral edema. Local and systemic inflammatory reactions are a major component of the IS pathophysiology and can significantly aggravate brain tissue damage. Intracerebral inflammatory processes following IS have been well studied. Until now, less is known about systemic inflammatory responses and IS consequences apart from a frequently observed post-IS immunosuppression. Here, we provide a hypothesis of a crosstalk between systemic acute phase response (APR), IP and potential secondary brain damage during acute and subacute IS stages supported by preliminary experimental data. Alterations of the acute phase proteins (APPs) C-reactive protein and lipopolysaccharide-binding protein and serum level changes of antibodies directed against Escherichia coli -cell extract antigen (IgA-, IgM-, and IgG-anti- E. coli ) were investigated at 1, 2, and 7 days following IS in ten male sheep. We found an increase of both APPs as well as a decrease of all anti- E. coli antibodies within 48 h following IS. This may indicate an early systemic APR and increased IP, and underlines the importance of the increasingly recognized gut-brain axis and of intestinal antigen release for systemic immune responses in acute and subacute stroke stages., (Copyright © 2020 Ferrara, Zeisig, Pietsch, Rütten, Dreyer, Pieper, Schatzl, McLeod, Barthel, Boltze, Schrödl and Nitzsche.)

The metabolic response to endotoxemia closely mimics those seen in sepsis. Here, we show that the urinary excretion of the metabolite 2-hydroxyglutarate (2HG) is dramatically suppressed following lipopolysaccharide (LPS) administration in vivo , and in human septic patients. We further show that enhanced activation of the enzymes responsible for 2-HG degradation, D- and L-2-HGDH, underlie this effect. To determine the role of supplementation with 2HG, we carried out co-administration of LPS and 2HG. This co-administration in mice modulates a number of aspects of physiological responses to LPS, and in particular, protects against LPS-induced hypothermia. Our results identify a novel role for 2HG in endotoxemia pathophysiology, and suggest that this metabolite may be a critical diagnostic and therapeutic target for sepsis., (Copyright © 2020 Fitzpatrick, Lambden, Macias, Puthucheary, Pietsch, Mendil, McPhail and Johnson.)

We present herein investigations concerning the reactivity of B2pin2 and FBpin (pin = pinacolato) with respect to fluoride ions and NHCs (NHC = N-Heterocyclic Carbene) in order to understand better the role of added fluoride in our recently reported Ni-catalyzed C-F borylation process, to confirm the nature of the NHC-adducts of FBpin, and to explain the formation of the species detected as side products of the defluoroborylation reaction. We report the calculated gas phase fluoride ion affinities (FIA) of relevant boron species and demonstrate that the presence of one equivalent of added F- in solution will bind and activate B2pin2 (FIA = +247 kJ mol-1), but it will also bind even more effectively to FBpin (FIA = +287 kJ mol-1) formed in the reaction. We prepared the NHC-adducts of FBpin by two different methods and isolated and characterized several examples of the type FBpin·NHC (NHC = iPr2Im, 1; Me2Im, 2; MeiPrIm, 3; nPr2Im, 4; Mes2Im 5). The salts [pinBF2][NMe4] 6, as well as the significantly more soluble analogue [pinBF2][NnBu4] 7, were also isolated and characterized, providing confirmation of the presence of the pinBF2 anion in various stoichiometric and catalytic borylation reactions.

Intracerebral hemorrhage (ICH) is an important stroke subtype, but preclinical research is limited by a lack of translational animal models. Large animal models are useful to comparatively investigate key pathophysiological parameters in human ICH. To (i) establish an acute model of moderate ICH in adult sheep and (ii) an advanced neuroimage processing pipeline for automatic brain tissue and hemorrhagic lesion determination; 14 adult sheep were assigned for stereotactically induced ICH into cerebral white matter under physiological monitoring. Six hours after ICH neuroimaging using 1.5T MRI including structural as well as perfusion and diffusion, weighted imaging was performed before scarification and subsequent neuropathological investigation including immunohistological staining. Controlled, stereotactic application of autologous blood caused a space-occupying intracerebral hematoma of moderate severity, predominantly affecting white matter at 5 h post-injection. Neuroimage post-processing including lesion probability maps enabled automatic quantification of structural alterations including perilesional diffusion and perfusion restrictions. Neuropathological and immunohistological investigation confirmed perilesional vacuolation, axonal damage, and perivascular blood as seen after human ICH. The model and imaging platform reflects key aspects of human ICH and enables future translational research on hematoma expansion/evacuation, white matter changes, hematoma evacuation, and other aspects.

Forest biomass is an essential indicator for monitoring the Earth's ecosystems and climate. It is a critical input to greenhouse gas accounting, estimation of carbon losses and forest degradation, assessment of renewable energy potential, and for developing climate change mitigation policies such as REDD+, among others. Wall-to-wall mapping of aboveground biomass (AGB) is now possible with satellite remote sensing (RS). However, RS methods require extant, up-to-date, reliable, representative and comparable in situ data for calibration and validation. Here, we present the Forest Observation System (FOS) initiative, an international cooperation to establish and maintain a global in situ forest biomass database. AGB and canopy height estimates with their associated uncertainties are derived at a 0.25 ha scale from field measurements made in permanent research plots across the world's forests. All plot estimates are geolocated and have a size that allows for direct comparison with many RS measurements. The FOS offers the potential to improve the accuracy of RS-based biomass products while developing new synergies between the RS and ground-based ecosystem research communities.

Background: Autologous chondrocyte implantation (ACI) and microfracture are established treatments for large, full-thickness cartilage defects, but there is still a need to expand the clinical and health economic knowledge of these procedures., Purpose: To confirm the noninferiority of ACI compared with microfracture., Study Design: Randomized controlled trial; Level of evidence, 2., Methods: Patients were randomized to be treated with matrix-associated ACI using spheroid technology (n = 52) or microfracture (n = 50). Both procedures followed standard methods. Patients were assessed by the Knee injury and Osteoarthritis Outcome Score (KOOS), MOCART (magnetic resonance observation of cartilage repair tissue) scoring system, Bern score, modified Lysholm score, International Cartilage Repair Society (ICRS) rating (histological and immunochemical scoring after rebiopsy 24 months after implantation), and International Knee Documentation Committee (IKDC) examination form. The main assessments were conducted 24 months after study treatment., Results: In the primary intention-to-treat analysis, the overall KOOS score for both ACI and microfracture yielded a statistically significant improvement relative to baseline. According to the between-group analysis, ACI passed the test of noninferiority compared with microfracture; thus, the primary goal of the study was achieved. The KOOS subscores yielded the same qualitative results as the overall KOOS score (ie, for each of these, noninferiority was demonstrated), and in 1 case (Activities of Daily Living subscore), the threshold for superiority was passed. The subgroup analyses did not yield any clear evidence of an association between treatment effect and any of the categories investigated (age, diagnosis, defect localization, sex). A histological analysis of biopsies from 16 patients (ACI: n = 9; microfracture: n = 7) suggested a better quality of repair in the patients treated with ACI., Conclusion: The efficacy of both ACI and microfracture was demonstrated with respect to both functional outcomes and morphological repair. The primary analysis confirmed the statistical hypothesis of the noninferiority of ACI, even for relatively small cartilage defects (1-4 cm 2 ) treated in this study, the indication for which microfracture is generally accepted as the standard of care. ACI showed significant superiority in the KOOS subscores of Activities of Daily Living at 24 months and Knee-related Quality of Life at 12 months., Registration: NCT01222559 (ClinicalTrials.gov identifier)., Competing Interests: One or more of the authors has declared the following potential conflict of interest or source of funding: The present study was funded by CO.DON. P.N. has received consulting fees from CO.DON and has received educational support from Aesculap, Arthrex, Geistlich, Moximed, Stryker, and Medi. W.Z. has received consulting fees from CO.DON. S.F. has received consulting fees from Arthrex and Bauerfeind. AOSSM checks author disclosures against the Open Payments Database (OPD). AOSSM has not conducted an independent investigation on the OPD and disclaims any liability or responsibility relating thereto.

Prior injury is a commonly identified risk factor for subsequent injury. However, a binary approach to classifying prior injury (i.e., yes/no) is commonly implemented and may constrain scientific findings, as it is possible that variations in the amount of time lost due to an injury will impact subsequent injury risk to differing degrees. Accordingly, this study investigated whether session availability, a surrogate marker of prior injury, influenced the risk of subsequent non-contact lower limb injury in Australian footballers. Data were collected from 62 male elite Australian footballers throughout the 2015, 2016, and 2017 Australian Football League seasons. Each athlete's participation status (i.e., full or missed/modified) and any injuries that occurred during training sessions/matches were recorded. As the focus of the current study was prior injury, any training sessions/matches that were missed due to reasons other than an injury (e.g., load management, illness and personal reasons) were removed from the data prior to all analyses. For every Monday during the in-season periods, session availability (%) in the prior 7, 14, 21, 28, 35, 42, 49, 56, 63, 70, 77, and 84 days was determined as the number of training sessions/matches fully completed (injury free) relative to the number of training sessions/matches possible in each window. Each variable was modeled using logistic regression to determine its impact on subsequent injury risk. Throughout the study period, 173 non-contact lower limb injuries that resulted in at least one missed/modified training session or match during the in-season periods occurred. Greater availability in the prior 7 days increased injury probabilities by up to 4.4%. The impact of session availability on subsequent injury risk diminished with expanding windows (i.e., availability in the prior 14 days through to the prior 84 days). Lesser availability in the prior 84 days increased injury probabilities by up to 14.1%, only when coupled with greater availability in the prior 7 days. Session availability may provide an informative marker of the impact of prior injury on subsequent injury risk and can be used by coaches and clinicians to guide the progression of training, particularly for athletes that are returning from long periods of injury.

This study investigates mental rotation performance of adolescent female dancers and soccer players in object-based and egocentric mental rotation tasks using human body stimuli. 60 young females, 30 soccer players, and 30 dancers (not twosome), completed a chronometric mental rotation task with object-based and egocentric transformation of male and female figures, which were displayed either in front or back view. During their sport-specific activity soccer-players and dancers very often have to adapt their movements to the movement of a partner or opponent, soccer-players especially in front view positions. While for soccer-players reaction time (RT) often is crucial for sporting success, dancers mainly focus on the accuracy of their movements. Therefore, we expect significantly faster RTs for soccer players for front view stimuli but no differences between soccer players and dancers for back view stimuli. The main result was that soccer-players showed a significantly shorter RT than dancers for stimuli presented in front view in object based and egocentric transformation. There was no such difference, when the stimuli were presented in the back view. Contrary to literature we didn't find significantly higher RTs and error rates for stimuli presented in front view compared to back view in general but only for egocentric transformations. The results of this study show that specific sports affect individual aspects of mental rotation performance.

This study investigates the mental rotation performance of right- and left-handers on a chronometric mental rotation task. Hand preference as well as different types of hand performance were measured. Previous studies have reported slightly better mental rotation performance in right-handers compared with left-handers. However, different forms of hand performance were not considered in these studies. In this research, 61 participants completed a chronometric mental rotation task with cube figures. Handedness was measured using the Edinburgh Handedness Inventory, finger tapping, grip strength and the Grooved Pegboard Test. Our results showed that right-handers do not generally show faster reaction times than left-handers on mental rotation tasks, when handedness is measured using a typical handedness questionnaire. However, using a handedness measure of visuo-motor control and manipulation skill, we found that individuals who solved the task faster with their right hand significantly outperformed participants who solved the same test faster with their left hand., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Introduction: The use of Kinesio tape (KT) to improve proprioception is a matter of considerable debate. In comparison, the rupture of the anterior cruciate ligament is a sufficiently well-investigated injury with a proven compromise of proprioception. The objective of the present study was to assess a supportive effect on proprioception after KT application, taking the anterior cruciate ligament (ACL) rupture as an example., Materials and Methods: Forty-eight patients who had suffered an ACL rupture, confirmed clinically and by magnetic resonance imaging, and who were treated conservatively or were awaiting surgery were included in this study. In all patients, a gait analysis was performed on the affected leg before and after KT application. In addition, the IKDC score, the Lysholm score, stability using the Rolimeter, and the angle reproduction test were determined., Results: Thirty-nine men and nine women who had had an ACL rupture for at least 3 weeks were included in the study. Significant improvements were achieved on the affected knee joint for the gait analysis parameters touchdown and unrolling, cadence, stability and stance phase as well as an extension of the hip joint. The Lysholm score improved from 79.3 to 85.8 (p < 0.001) and the IKDC score from 60.2 to 71.3 points (p < 0.001). Significant improvements were achieved in the Rolimeter and angle reproduction test., Conclusions: The use of KT has a positive effect on proprioception in patients with an anterior cruciate ligament rupture. Therefore, the application may improve gait pattern as well as the subjective function of the affected knee joint.

Animals require an immediate response to oxygen availability to allow rapid shifts between oxidative and glycolytic metabolism. These metabolic shifts are highly regulated by the HIF transcription factor. The factor inhibiting HIF (FIH) is an asparaginyl hydroxylase that controls HIF transcriptional activity in an oxygen-dependent manner. We show here that FIH loss increases oxidative metabolism, while also increasing glycolytic capacity, and that this gives rise to an increase in oxygen consumption. We further show that the loss of FIH acts to accelerate the cellular metabolic response to hypoxia. Skeletal muscle expresses 50-fold higher levels of FIH than other tissues: we analyzed skeletal muscle FIH mutants and found a decreased metabolic efficiency, correlated with an increased oxidative rate and an increased rate of hypoxic response. We find that FIH, through its regulation of oxidation, acts in concert with the PHD/vHL pathway to accelerate HIF-mediated metabolic responses to hypoxia., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)

This study investigates the influence of specific soccer training with the non-dominant leg on mental rotation performance of 20 adolescent soccer players between 10 and 11 years of age. While the experimental group performed soccer specific tasks only with the non-dominant foot once a week for 10 weeks, the control group absolved the same exercises with the dominant foot for the same period of time. Both groups performed a mental rotation task and shot, dribbling and ball control tests before and after the 10 week intervention. The most relevant result was that the experimental group showed a significantly larger increase in mental rotation ability than the control group.

We analysed the ligand-based activation mechanism of the prostate-specific G-protein coupled receptor (PSGR), which is an olfactory receptor that mediates cellular growth in prostate cancer cells. Furthermore, it is an olfactory receptor with a known chemically near identic antagonist/agonist pair, α- and β-ionone. Using a combined theoretical and experimental approach, we propose that this receptor is activated by a ligand-induced rearrangement of a protein-internal hydrogen bond network. Surprisingly, this rearrangement is not induced by interaction of the ligand with the network, but by dynamic van der Waals contacts of the ligand with the involved amino acid side chains, altering their conformations and intraprotein connectivity. Ligand recognition in this GPCR is therefore highly stereo selective, but seemingly lacks any ligand recognition via polar contacts. A putative olfactory receptor-based drug design scheme will have to take this unique mode of protein/ligand action into account.

In this detailed study we report on the structures of the widely employed diboron(4) compounds bis(pinacolato)diboron (B 2 pin 2 ) and bis(neopentyl glycolato)diboron (B 2 neop 2 ), as well as bis(ethylene glycolato)diboron (B 2 eg 2 ) and tetrakis(dimethylamino)diboron (B 2 (NMe 2 ) 4 ), and their reactivity, along with that of bis(catecholato)diboron (B 2 cat 2 ) with backbone saturated and backbone unsaturared N-heterocyclic carbenes (NHCs) of different steric demand. Depending on the nature of the diboron(4) compound and the NHC used, Lewis-acid/Lewis-base adducts or NHC ring-expansion products stemming from B-B and C-N bond activation have been observed. The corresponding NHC adducts and NHC ring-expanded products were isolated and characterised via solid-state and solution NMR spectroscopy and X-ray diffraction. In general, we observed B-B bond and C-N bond activation at low temperature for B 2 eg 2 , at room temperature for B 2 neop 2 and at higher temperature for B 2 cat 2 . The reactivity strongly depends on steric effects of the NHCs and the diboron(4) compounds, as well as on the corresponding Lewis-basicity and Lewis-acidity. Our results provide profound insight into the chemistry of these diboron(4) reagents with the nowadays ubiquitous NHCs, the stability of the corresponding NHC adducts and on B-B bond activation using Lewis-bases in general. We demonstrate that B-B bond activation may be triggered even at temperatures as low as -40 °C to -30 °C without any metal species involved. For example, the reactions of B 2 eg 2 with sterically less demanding NHCs such as Me 2 Im Me and iPr 2 Im in solution led to the corresponding ring-expanded products at low temperatures. Furthermore, boronium [L 2 B(OR) 2 ] + and borenium [LB(OR) 2 ] + cations have been observed from the reaction of the bis-borate B 2 eg 3 with the NHCs iPr 2 Im and Me 2 Im Me , which led to the conclusion that the activation of bis-borates with NHCs (or Lewis-bases in general) might be a facile and simple route to access such species.

We report the reactions of catecholborane (HBcat; 1) with unsaturated and saturated NHCs as well as CAAC(Me) . Mono-NHC adducts of the type HBcat⋅NHC (NHC=nPr2 Im, iPr2 Im, iPr2 Im(Me) , and Dipp2 Im) were obtained by stoichiometric reactions of HBcat with the unsaturated NHCs. The reaction of CAAC(Me) with HBcat yielded the B-H activated product CAAC(Me) (H)Bcat via insertion of the carbine-carbon atom into the B-H bond. The saturated NHC Dipp2 SIm reacted in a 2:2 ratio yielding an NHC ring-expanded product at room temperature forming a six-membered -B-C=N-C=C-N- ring via C-N bond cleavage and further migration of the hydrides from two HBcat molecules to the former carbene-carbon atom., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Biodiversity loss is mainly driven by human activity. While concern grows over the fate of hot spots of biodiversity, contemporary species losses still prevail in industrialized nations. Therefore, strategies were formulated to halt or reverse the loss, driven by evidence for its value for ecosystem services. Maintenance of the latter through conservation depends on correctly identified species. To this aim, the German Federal Ministry of Education and Research is funding the GBOL project, a consortium of natural history collections, botanic gardens, and universities working on a barcode reference database for the country's fauna and flora. Several noticeable findings could be useful for future campaigns: (i) validating taxon lists to serve as a taxonomic backbone is time-consuming, but without alternative; (ii) offering financial incentives to taxonomic experts, often citizen scientists, is indispensable; (iii) completion of the libraries for widespread species enables analyses of environmental samples, but the process may not hold pace with technological advancements; (iv) discoveries of new species are among the best stories for the media; (v) a commitment to common data standards and repositories is needed, as well as transboundary cooperation between nations; (vi) after validation, all data should be published online via the BOLD to make them searchable for external users and to allow cross-checking with data from other countries.

N-Heterocyclic carbenes (NHCs) are widely used ligands and reagents in modern inorganic synthesis as well as in homogeneous catalysis and organocatalysis. However, NHCs are not always innocent bystanders. In the last few years, more and more examples were reported of reactions of NHCs with main-group elements which resulted in modification of the NHC. Many of these reactions lead to ring expansion and the formation of six-membered heterocyclic rings involving insertion of the heteroatom into the C-N bond and migration of hydrides, phenyl groups or boron-containing fragments. Furthermore, a few related NHC rearrangements were observed some decades ago. In this Perspective, we summarise the history of NHC ring expansion reactions from the 1960s till the present.

The killifish Nothobranchius furzeri is the shortest-lived vertebrate that can be bred in the laboratory. Its rapid growth, early sexual maturation, fast aging, and arrested embryonic development (diapause) make it an attractive model organism in biomedical research. Here, we report a draft sequence of its genome that allowed us to uncover an intra-species Y chromosome polymorphism representing-in real time-different stages of sex chromosome formation that display features of early mammalian XY evolution "in action." Our data suggest that gdf6Y, encoding a TGF-β family growth factor, is the master sex-determining gene in N. furzeri. Moreover, we observed genomic clustering of aging-related genes, identified genes under positive selection, and revealed significant similarities of gene expression profiles between diapause and aging, particularly for genes controlling cell cycle and translation. The annotated genome sequence is provided as an online resource (http://www.nothobranchius.info/NFINgb)., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Acute kidney injury in mammals, which is caused by cardiovascular diseases or the administration of antibiotics with nephrotoxic side-effects is a life-threatening disease, since loss of nephrons is irreversible in mammals. In contrast, fish are able to generate new nephrons even in adulthood and thus provide a good model to study renal tubular regeneration., Results: Here, we investigated the early response after gentamicin-induced renal injury, using the short-lived killifish Nothobranchius furzeri. A set of microRNAs was differentially expressed after renal damage, among them miR-21, which was up-regulated. A locked nucleic acid-modified antimiR-21 efficiently knocked down miR-21 activity and caused a lag in the proliferative response, enhanced apoptosis and an overall delay in regeneration. Transcriptome profiling identified apoptosis as a process that was significantly affected upon antimiR-21 administration. Together with functional data this suggests that miR-21 acts as a pro-proliferative and anti-apoptotic factor in the context of kidney regeneration in fish. Possible downstream candidate genes that mediate its effect on proliferation and apoptosis include igfbp3 and fosl1, among other genes., Conclusion: In summary, our findings extend the role of miR-21 in the kidney. For the first time we show its functional involvement in regeneration indicating that fast proliferation and reduced apoptosis are important for efficient renal tubular regeneration.

Superhalogens belong to a class of molecules that not only mimic the chemistry of halogen atoms but also possess electron affinities that are much larger than that of chlorine, the element with the highest electron affinity in the periodic table. Using BO2 as an example and the synergy between density functional theory-based calculations and photoelectron spectroscopy experiments we demonstrate another unusual property of superhalogens. Unlike halogens, whose ability to accept an electron falls upon dimerization, B2O4, the dimer of BO2, has an electron affinity larger than that of the BO2 building block. This ability of (BO2)2 and subsequent, higher oligomers (BO2)n (n = 3 and 4), to retain their superhalogen characteristics can be traced to the enhanced bonding interactions between oxygen and boron atoms and due to the delocalization of the charge of the extra-electron over the terminal oxygen atoms. These results open the door to the design and synthesis of a new class of metal-free highly negative ions with potential for novel applications.

Mutations in Wilms' tumor 1 (WT1) cause a wide spectrum of renal manifestations, eventually leading to end-stage kidney failure. Insufficient understanding of WT1's molecular functions in kidney development has hampered efficient therapeutic applications for WT1-associated diseases. Recently, the generation and characterization of mouse models and application of multiple state-of-the-art approaches have significantly expanded our understanding of the molecular mechanisms of how WT1 mutations lead to kidney failure. Here, we discuss the WT1 binding consensus and illustrate the major roles of WT1 in different cell populations in kidney biology. WT1 controls metanephric mesenchyme (MM) self-renewal and proliferation mainly by regulating FGF and BMP-pSMAD signaling pathways as well as Sall1 and Pax2, encoding key transcription factors; WT1 drives MM differentiation and mesenchyme-epithelial transition by targeting Fgf8 and Wnt4; WT1 defines podocyte identity by activation of other podocyte-specific transcription factors, including Mafb, Lmx1b, FoxC2, and Tcf21. These factors potentially cooperate with WT1 regulating the expression of components and regulators of the cytoskeleton for establishing podocyte polarity, slit diaphragm structure, and focal adhesion to the glomerular basement membrane. Understanding of WT1's function in kidney biology including WT1-regulated pathways will give insights that will eventually help therapeutic applications.

The Wilms' tumor suppressor gene 1 (WT1) encodes a zinc finger transcription factor. Mutation of WT1 in humans leads to Wilms' tumor, a pediatric kidney tumor, or other kidney diseases, such as Denys-Drash and Frasier syndromes. We showed previously that inactivation of WT1 in podocytes of adult mice results in proteinuria, foot process effacement, and glomerulosclerosis. However, the WT1-dependent transcriptional network regulating podocyte development and maintenance in vivo remains unknown. Here, we performed chromatin immunoprecipitation followed by high-throughput sequencing with glomeruli from wild-type mice. Additionally, we performed a cDNA microarray screen on an inducible podocyte-specific WT1 knockout mouse model. By integration of cistromic and transcriptomic analyses, we identified the WT1 targetome in mature podocytes. To further analyze the function and targets of WT1 in podocyte maturation, we used an Nphs2-Cre model, in which WT1 is deleted during podocyte differentiation. These mice display anuria and kidney hemorrhage and die within 24 hours after birth. To address the evolutionary conservation of WT1 targets, we performed functional assays using zebrafish as a model and identified Nphs2, Mafb, and Magi2 as novel WT1 target genes required for podocyte development. Our data also show that both Mafb and Magi2 are required for normal development of the embryonic zebrafish kidney. Collectively, our work provides insights into the transcriptional networks controlled by WT1 and identifies novel WT1 target genes that mediate the function of WT1 in podocyte differentiation and maintenance., (Copyright © 2015 by the American Society of Nephrology.)

We report the isolation and detailed structural characterization, by solid-state and solution NMR spectroscopy, of the neutral mono- and bis-NHC adducts of bis(catecholato)diboron (B2 cat2 ). The bis-NHC adduct undergoes thermally induced rearrangement, forming a six-membered -B-C=N-C=C-N-heterocyclic ring via C-N bond cleavage and ring expansion of the NHC, whereas the mono-NHC adduct is stable. Bis(neopentylglycolato)diboron (B2 neop2 ) is much more reactive than B2 cat2 giving a ring expanded product at room temperature, demonstrating that ring expansion of NHCs can be a very facile process with significant implications for their use in catalysis., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Lewis base adducts of tetra-alkoxy diboron compounds, in particular bis(pinacolato)diboron (B2 pin2 ), have been proposed as the active source of nucleophilic boryl species in metal-free borylation reactions. We report the isolation and detailed structural characterization (by solid-state and solution NMR spectroscopy and X-ray crystallography) of a series of anionic adducts of B2 pin2 with hard Lewis bases, such as alkoxides and fluoride. The study was extended to alternative Lewis bases, such as acetate, and other diboron reagents. The B(sp(2) )-B(sp(3) ) adducts exhibit two distinct boron environments in the solid-state and solution NMR spectra, except for [(4-tBuC6 H4 O)B2 pin2 ](-) , which shows rapid site exchange in solution. DFT calculations were performed to analyze the stability of the adducts with respect to dissociation. Stoichiometric reaction of the isolated adducts with two representative series of organic electrophiles-namely, aryl halides and diazonium salts-demonstrate the relative reactivities of the anionic diboron compounds as nucleophilic boryl anion sources., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The Wilms tumor suppressor gene Wt1 encodes a zinc finger transcription factor that is essential for development of multiple organs including kidneys, gonads, spleen and heart. In mammals Wt1 comprises 10 exons with two characteristic splicing events: inclusion or skipping of exon 5 and alternative usage of two splice donor sites between exons 9 and 10. Most fish including zebrafish and medaka possess two wt1 paralogs, wt1a and wt1b, both lacking exon 5. Here we have characterized wt1 in guppy, platyfish and the short-lived African killifish Nothobranchius furzeri. All fish except zebrafish show alternative splicing of exon 4 of wt1a but not of wt1b with the wt1a(-exon 4) isoform being the predominant splice variant. With regard to function, Wt1a(+exon 4) showed less dimerization but stimulated transcription more effectively than the Wt1a(-exon 4) isoform. A specific knockdown of wt1a exon 4 in zebrafish was associated with anomalies in kidney development demonstrating a physiological function for Wt1a exon 4. Interestingly, alternative splicing of exon 4 seems to be an early evolutionary event as it is observed in the single wt1 gene of the sturgeon, a species that has not gone through teleost-specific genome duplication., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Objectives: To identify a molecular profile for schizophrenia using post-mortem pituitaries from schizophrenia and control subjects., Methods: Molecular profiling analysis of pituitaries from schizophrenia (n = 14) and control (n = 15) subjects was carried out using a combination of liquid chromatography tandem mass spectrometry (LC-MS(E)), multiplex analyte profiling (MAP), two-dimensional difference gel electrophoresis (2D-DIGE) and Western blot analysis., Results: This led to identification of differentially expressed molecules in schizophrenia patients including hypothalamic-pituitary-adrenal axis-associated constituents such as cortisol, pro-adrenocorticotropic hormone, arginine vasopressin precursor, agouti-related protein, growth hormone, prolactin and secretagogin, as well as molecules associated with lipid transport and metabolism such as apolipoproteins A1, A2, C3 and H. Altered levels of secretagogin in serum from a cohort of living first onset schizophrenia patients were also detected, suggesting disease association and illustrating the potential for translating some components of this molecular profile to serum-based assays., Conclusions: Future studies on the molecules identified here may lead to new insights into schizophrenia pathophysiology and pave the way for translation of novel diagnostics for use in a clinical setting.

Previous studies have found that some first onset schizophrenia patients show signs of impaired insulin signaling. Also, epidemiological studies have shown that periods of suboptimal nutrition including protein deficiencies during pregnancy can lead to increased incidence of metabolic conditions and psychiatric disorders in the offspring. For these reasons, we have carried out a molecular profiling analysis of blood serum and brain tissues from adult offspring produced by the maternal low protein (LP) rat model. The results showed similar changes to those seen in schizophrenia. Multiplex immunoassay profiling identified changes in the levels of insulin, adiponectin, and leptin along with alterations in inflammatory and vascular system-related proteins such as osteopontin, macrophage colony-stimulating factor 1, and vascular cell adhesion molecule 1. LC-MS(E) proteomic profiling showed that glutamatergic pathways were altered in frontal cortex, while signaling pathways and cytoskeletal proteins involved in hormonal secretion and synaptic remodeling were altered in the hypothalamus. Taken together, these studies indicate that the LP rat model recapitulates several pathophysiological attributes seen in schizophrenia patients. We propose that the LP model may have utility for drug discovery efforts, especially to identify compounds that modulate the metabolic and glutamatergic systems., (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)