Search

SPARC is a transiently expressed extracellular matrix-binding protein that alters cell shape and regulates endothelial cell proliferation in vitro. In this study, we show that SPARC mRNA and protein are synthesized by endothelial cells during angiogenesis in vivo. SPARC and peptides derived from a cationic region of the protein (amino acids 113-130) stimulated the formation of endothelial cords in vitro; moreover, these peptides stimulated angiogenesis in vivo. Mapping of the active domain demonstrated that the sequence KGHK was responsible for most of the angiogenic activity; substitution of the His residue decreased the effect. We found that proteolysis of SPARC provided a source of KGHK, GHK, and longer peptides that contained these sequences. Although the Cu(2+)-GHK complex had been identified as a mitogen/morphogen in normal human plasma, we found KGHK and longer peptides to be potent stimulators of angiogenesis. SPARC113-130 and KGHK were shown to bind Cu2+ with high affinity; however, previous incubation with Cu2+ was not required for the stimulatory activity. Since a peptide from a second cationic region of SPARC (SPARC54-73) also bound Cu2+ but had no effect on angiogenesis, the angiogenic activity appeared to be sequence specific and independent of bound Cu2+. Thus, specific degradation of SPARC, a matrix-associated protein expressed by endothelial cells during vascular remodeling, releases a bioactive peptide or peptides, containing the sequence (K)GHK, that could regulate angiogenesis in vivo.

Hypoxia-inducible factors (HIFs) are broadly expressed in human cancers, and HIF1α and HIF2α were previously suspected to promote tumour progression through largely overlapping functions. However, this relatively simple model has now been challenged in light of recent data from various approaches that reveal unique and sometimes opposing activities of these HIFα isoforms in both normal physiology and disease. These effects are mediated in part through the regulation of unique target genes, as well as through direct and indirect interactions with important oncoproteins and tumour suppressors, including MYC and p53. As HIF inhibitors are currently undergoing clinical evaluation as cancer therapeutics, a more thorough understanding of the unique roles performed by HIF1α and HIF2α in human neoplasia is warranted. [ABSTRACT FROM AUTHOR]

Hypoxia-inducible factor (HIF)-1alpha is activated on exposure to bacterial pathogens and regulates the innate immune functions of phagocytes. We show here that the HIF-1alpha agonist mimosine can boost the capacity of human phagocytes and whole blood to kill the leading pathogen Staphylococcus aureus in a dose-dependent fashion and reduce the lesion size in a murine model of S. aureus skin infection. This provides the first proof of principle for a novel approach to the treatment of bacterial infection by pharmacologically augmenting the host phagocytic function. Copyright © 2008 Infectious Diseases Society of America [ABSTRACT FROM AUTHOR]

Background: Concussions are a prevalent health concern, affecting millions of individuals in the United States yearly. Concussion symptomology overlaps with other conditions making diagnosis and management particularly challenging, particularly among individuals with ADHD. ADHD is a neurodevelopmental disorder that affects a significant portion of the adult population., Objective: To enhance our understanding of symptom resolution and identify factors that could influence concussion care management, the purpose of the current study was to examine the recovery trajectory of adults diagnosed with both ADHD and concussion compared to the trajectory of adults who have experienced a concussion but do not have an ADHD diagnosis., Methods: The study included a retrospective chart review of patients treated for concussion at a concussion specialty clinic., Results: The findings of the study revealed no significant differences in history of concussion or recovery time between patients with and without ADHD. It was observed that patients with ADHD had a higher prevalence of anxiety, depression, and other mental health conditions., Conclusions: The findings of the current study underscore the need for additional research to guide the care of patients diagnosed with both ADHD and concussion., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Brain microvascular dysfunction is an important feature of Alzheimer's disease (AD). To better understand the brain microvascular molecular signatures of AD, we processed and analyzed isolated human brain microvessels by data-independent acquisition liquid chromatography with tandem mass spectrometry (DIA LC-MS/MS) to generate a quantitative dataset at the peptide and protein level. Brain microvessels were isolated from parietal cortex grey matter using protocols that preserve viability for downstream functional studies. Our cohort included 23 subjects with clinical and neuropathologic concordance for Alzheimer's disease, and 21 age-matched controls. In our analysis, we identified 168 proteins whose abundance was significantly increased, and no proteins that were significantly decreased in AD. The most highly increased proteins included amyloid beta, tau, midkine, SPARC related modular calcium binding 1 (SMOC1), and fatty acid binding protein 7 (FABP7). Additionally, Gene Ontology (GO) enrichment analysis identified the enrichment of increased proteins involved in cellular detoxification and antioxidative responses. A systematic evaluation of protein functions using the UniProt database identified groupings into common functional themes including the regulation of cellular proliferation, cellular differentiation and survival, inflammation, extracellular matrix, cell stress responses, metabolism, coagulation and heme breakdown, protein degradation, cytoskeleton, subcellular trafficking, cell motility, and cell signaling. This suggests that AD brain microvessels exist in a stressed state of increased energy demand, and mount a compensatory response to ongoing oxidative and cellular damage that is associated with AD. We also used public RNAseq databases to identify cell-type enriched genes that were detected at the protein level and found no changes in abundance of these proteins between control and AD groups, indicating that changes in cellular composition of the isolated microvessels were minimal between AD and no-AD groups. Using public data, we additionally found that under half of the proteins that were significantly increased in AD microvessels had concordant changes in brain microvascular mRNA, implying substantial discordance between gene and protein levels. Together, our results offer novel insights into the molecular underpinnings of brain microvascular dysfunction in AD., (© 2024. The Author(s).)

A thorough evaluation of the quality, reproducibility, and variability of bottom-up proteomics data is necessary at every stage of a workflow, from planning to analysis. We share vignettes applying adaptable quality control (QC) measures to assess sample preparation, system function, and quantitative analysis. System suitability samples are repeatedly measured longitudinally with targeted methods, and we share examples where they are used on three instrument platforms to identify severe system failures and track function over months to years. Internal QCs incorporated at the protein and peptide levels allow our team to assess sample preparation issues and to differentiate system failures from sample-specific issues. External QC samples prepared alongside our experimental samples are used to verify the consistency and quantitative potential of our results during batch correction and normalization before assessing biological phenotypes. We combine these controls with rapid analysis (Skyline), longitudinal QC metrics (AutoQC), and server-based data deposition (PanoramaWeb). We propose that this integrated approach to QC is a useful starting point for groups to facilitate rapid quality control assessment to ensure that valuable instrument time is used to collect the best quality data possible. Data are available on Panorama Public and ProteomeXchange under the identifier PXD051318.

Objective: To identify (1) university educators' perceptions of academic adjustments (AA), and (2) if teaching experience correlated with AA perceptions following concussion. Participants: Two hundred twenty educators. Methods: University educators were invited to complete a survey containing four subsections; this manuscript focuses on AA following concussion. Objective 1 was descriptive; we conducted spearman's rho correlations between years of teaching experience and AA perceptions to address objective 2. Results: Educators were moderately familiar with AA but were not confident in their knowledge about AA following concussion. Participants who provided AA following concussion most often allowed excused absences and extra time for exams/assignments. There were no significant relationships between teaching experience and perceptions of AA. Conclusions: University educators largely feel unprepared to provide or recommend AA following concussion but had favorable AA perceptions following concussion. Standardized policies or referral sites within the university system may be warranted to improve post-concussion AA.

Sex has a major effect on the metabolome. However, we do not yet understand the degree to which these quantitative sex differences in metabolism are associated with anatomical dimorphism and modulated by sex-specific tissues. In the fruit fly, Drosophila melanogaster , knocking out the doublesex ( dsx ) gene gives rise to adults with intermediate sex characteristics. Here we sought to determine the degree to which this key node in sexual development leads to sex differences in the fly metabolome. We measured 91 metabolites across head, thorax and abdomen in Drosophila , comparing the differences between distinctly sex-dimorphic flies with those of reduced sexual dimorphism: dsx null flies. Notably, in the reduced dimorphism flies, we observed a sex difference in only 1 of 91 metabolites, kynurenate, whereas 51% of metabolites (46/91) were significantly different between wildtype XX and XY flies in at least one tissue, suggesting that dsx plays a major role in sex differences in fly metabolism. Kynurenate was consistently higher in XX flies in both the presence and absence of functioning dsx . We observed tissue-specific consequences of knocking out dsx . Metabolites affected by sex were significantly enriched in branched chain amino acid metabolism and the mTOR pathway. This highlights the importance of considering variation in genes that cause anatomical sexual dimorphism when analyzing sex differences in metabolic profiles and interpreting their biological significance.

Several metabolites have been shown to have independent and at times unexpected biological effects outside of their metabolic pathways. These include succinate, lactate, fumarate, and 2-hydroxyglutarate. 2-Hydroxybutyrate (2HB) is a byproduct of endogenous cysteine synthesis, produced during periods of cellular stress. 2HB rises acutely after exercise; it also rises during infection and is also chronically increased in a number of metabolic disorders. We show here that 2HB inhibits branched-chain aminotransferase enzymes, which in turn triggers a SIRT4-dependent shift in the compartmental abundance of protein ADP-ribosylation. The 2HB-induced decrease in nuclear protein ADP-ribosylation leads to a C/EBPβ-mediated transcriptional response in the branched-chain amino acid degradation pathway. This response to 2HB exposure leads to an improved oxidative capacity in vitro. We found that repeated injection with 2HB can replicate the improvement to oxidative capacity that occurs following exercise training. Together, we show that 2-HB regulates fundamental aspects of skeletal muscle metabolism., Competing Interests: BW, ML, EM, PC, VE, CB, CA, SS, EM, PK, JR, HR, JL, RJ No competing interests declared, (© 2023, Wadsworth et al.)

T cells must adapt to variations in tissue microenvironments; these adaptations include the degree of oxygen availability. The hypoxia-inducible factor (HIF) transcription factors control much of this adaptation, and thus regulate many aspects of T cell activation and function. The HIFs are in turn regulated by oxygen-dependent hydroxylases: both the prolyl hydroxylases (PHDs) which interact with the VHL tumour suppressor and control HIF turnover, and the asparaginyl hydroxylase known as the Factor inhibiting HIF (FIH), which modulates HIF transcriptional activity. To determine the role of this latter factor in T cell function, we generated T cell-specific FIH knockout mice. We found that FIH regulates T cell fate and function in a HIF-dependent manner and show that the effects of FIH activity occur predominantly at physiological oxygen concentrations. T cell-specific loss of FIH boosts T cell cytotoxicity, augments T cell expansion in vivo , and improves anti-tumour immunotherapy in mice. Specifically inhibiting FIH in T cells may therefore represent a promising strategy for cancer immunotherapy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Bargiela, Cunha, Veliça, Krause, Brice, Barbieri, Gojkovic, Foskolou, Rundqvist and Johnson.)

We show that a recently-derived model for the propagation of nonlinear waves in the atmosphere admits undular bores as travelling-wave solutions. These solutions represent waves consisting of a damped oscillation behind a front that is preceded by a uniform breeze-type flow. The generation of such wave profiles requires a jump in the heat source across the leading front of the wave, a feature that is consistent with observations., Competing Interests: Conflict of interestWe declare we have no competing interests., (© The Author(s) 2023.)

T cell function and fate can be influenced by several metabolites: in some cases, acting through enzymatic inhibition of α-ketoglutarate-dependent dioxygenases, in others, through post-translational modification of lysines in important targets. We show here that glutarate, a product of amino acid catabolism, has the capacity to do both, and has potent effects on T cell function and differentiation. We found that glutarate exerts those effects both through α-ketoglutarate-dependent dioxygenase inhibition, and through direct regulation of T cell metabolism via glutarylation of the pyruvate dehydrogenase E2 subunit. Administration of diethyl glutarate, a cell-permeable form of glutarate, alters CD8 + T cell differentiation and increases cytotoxicity against target cells. In vivo administration of the compound is correlated with increased levels of both peripheral and intratumoural cytotoxic CD8 + T cells. These results demonstrate that glutarate is an important regulator of T cell metabolism and differentiation with a potential role in the improvement of T cell immunotherapy., (© 2023. The Author(s).)

2-Hydroxyglutarate (2HG) is a byproduct of the tricarboxylic acid (TCA) cycle and is readily detected in the tissues of healthy individuals. 2HG is found in two enantiomeric forms: S-2HG and R-2HG. Here, we investigate the differential roles of these two enantiomers in cluster of differentiation (CD)8 + T cell biology, where we find they have highly divergent effects on proliferation, differentiation, and T cell function. We show here an analysis of structural determinants that likely underlie these differential effects on specific α-ketoglutarate (αKG)-dependent enzymes. Treatment of CD8 + T cells with exogenous S-2HG, but not R-2HG, increased CD8 + T cell fitness in vivo and enhanced anti-tumor activity. These data show that S-2HG and R-2HG should be considered as two distinct and important actors in the regulation of T cell function., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

This study aimed to examine the effect of football shoes with different collar types on ankle and knee kinematic and kinetics features during 45° and 135° side-step cutting tasks. Fifteen healthy college football players volunteered for the study. Each participant was instructed to perform side-step cutting tasks with high, low, and no collar football shoes. The kinematic and ground reaction force data were measured using a Vicon motion capture system and a Kistler force plate, respectively. Two-way MANOVAs with repeated measures were used to examine the effect of shoe collar type and task conditions. There were no interaction effects. The high collar football shoe showed decreased ankle range of motion in the sagittal plane ( p  = 0.010) and peak ankle external rotation moment ( p  = 0.009) compared to the no collar football shoe. The high ( p  = 0.025) and low ( p  = 0.029) collar football shoes presented greater peak ankle external rotation angles than the no collar football shoe. These results imply that football shoes with high collars made of high intensity knitted fabric could be used to restrict ankle joint movement, with potential implications for decreasing the risk of ankle sprain injuries in football players.

Introduction: Wearable accelerometry devices quantify on-field frequency and severity of head impacts to further improve sport safety. Commonly employed post-data collection cleaning techniques may affect these outcomes., Objective: Our purpose was to compare game impact rates and magnitudes between three different cleaning levels (Level-1: impacts recorded within start and end times, Level-2: impacts during pauses/breaks removed, Level-3: video verified) for male youth tackle football., Methods: Participants ( n  = 23, age = 10.9 ± 0.3 yrs, height = 150.0 ± 8.3 cm, mass = 41.6 ± 8.4 kg) wore Triax SIM-G sensors throughout Fall 2019. Impact rates, ratios (IRRs), and 95% confidence intervals (95%CI) were used to compare levels. Random-effects general linear models were used to compare peak linear acceleration (PLA;g) and angular velocity (PAV;rads/s)., Results: Level-1 resulted in higher impact rates (4.57; 95%CI = 4.14-5.05) compared to Level-2 (3.09; 95%CI = 2.80-3.42; IRR = 1.48; 95%CI = 1.34-1.63) and Level-3 datasets (2.56; 95%CI = 2.30-2.85; IRR = 1.78; 95%CI = 1.60-1.98). Level-2 had higher impact rates compared to Level-3 (1.21; 95%CI = 1.08-1.35). Level-1 resulted in higher PAV than Level-2 and Level-3 ( p  < 0.001) datasets. PLA did not differ across datasets ( p  = 0.296)., Conclusions: Head impact data should be filtered of pauses/breaks, and does not substantially differ outcome estimates compared to time-intensive video verification.

Oxygenation levels are a determinative factor in T cell function. Here, we describe how oxygen tensions sensed by mouse and human T cells at the moment of activation act to persistently modulate both differentiation and function. We found that in a protocol of CAR-T cell generation, 24 hr of low oxygen levels during initial CD8 + T cell priming is sufficient to enhance antitumour cytotoxicity in a preclinical model. This is the case even when CAR-T cells are subsequently cultured under high oxygen tensions prior to adoptive transfer. Increased hypoxia-inducible transcription factor (HIF) expression was able to alter T cell fate in a similar manner to exposure to low oxygen tensions; however, only a controlled or temporary increase in HIF signalling was able to consistently improve cytotoxic function of T cells. These data show that oxygenation levels during and immediately after T cell activation play an essential role in regulating T cell function., Competing Interests: PC, EM, LK, RH, DB, BW, LB, CB, IF, IB, PV, RJ No competing interests declared, (© 2023, Cunha et al.)