Kinase Suppressor of RAS 1 (KSR1) Maintains the Transformed Phenotype of BRAFV600E Mutant Human Melanoma Cells.

Kinase Suppressor of RAS 1 (KSR1) is a scaffolding protein for the RAS-RAF-MEK-ERK pathway, which is one of the most frequently altered pathways in human cancers. Previous results have shown that KSR1 has a critical role in mutant RAS-mediated transformation. Here, we examined the role of KSR1 in mutant BRAF transformation. We used CRISPR/Cas9 to knock out KSR1 in a BRAFV600E-transformed melanoma cell line. KSR1 loss produced a complex phenotype characterised by impaired proliferation, cell cycle defects, decreased transformation, decreased invasive migration, increased cellular senescence, and increased apoptosis. To decipher this phenotype, we used a combination of proteomic ERK substrate profiling, global protein expression profiling, and biochemical validation assays. The results suggest that KSR1 directs ERK to phosphorylate substrates that have a critical role in ensuring cell survival. The results further indicate that KSR1 loss induces the activation of p38 Mitogen-Activated Protein Kinase (MAPK) and subsequent cell cycle aberrations and senescence. In summary, KSR1 function plays a key role in oncogenic BRAF transformation.

The novel low molecular weight MYC antagonist MYCMI-6 inhibits proliferation and induces apoptosis in breast cancer cells.

Background The MYC oncogene is one of the most frequently altered driver genes in cancer. MYC is thus a potential target for cancer treatment as well as a biomarker for the disease. However, as a target for treatment, MYC has traditionally been regarded as "undruggable" or difficult to target. We set out to evaluate the efficacy of a novel MYC inhibitor known as MYCMI-6, which acts by preventing MYC from interacting with its cognate partner MAX. Methods MYCMI-6 response was assessed in a panel of breast cancer cell lines using MTT assays and flow cytometry. MYC gene amplification, mRNA and protein expression was analysed using the TCGA and METABRIC databases. Results MYCMI-6 inhibited cell growth in breast cancer cell lines with IC50 values varying form 0.3 µM to >10 µM. Consistent with its ability to decrease cell growth, MYCMI-6 was found to induce apoptosis in two cell lines in which growth was inhibited but not in two cell lines that were resistant to growth inhibition. Across all breast cancers, MYC was found to be amplified in 15.3% of cases in the TCGA database and 26% in the METABRIC database. Following classification of the breast cancers by their molecular subtypes, MYC was most frequently amplified and exhibited highest expression at both mRNA and protein level in the basal subtype. Conclusions Based on these findings, we conclude that for patients with breast cancer, anti-MYC therapy is likely to be most efficacious in patients with the basal subtype.

Long Covid brain fog: a neuroinflammation phenomenon?

Neuroinflammation is a process triggered by an attack on the immune system. Activation of microglia in response to an immune system challenge can lead to a significant impact on cognitive processes, such as learning, memory and emotional regulation. Long Covid is an ongoing problem, affecting an estimated 1.3 million people within the UK alone, and one of its more significant, and as yet unexplained, symptoms is brain fog. Here, we discuss the potential role of neuroinflammation in Long Covid cognitive difficulties. Inflammatory cytokines have been found to play a significant role in reductions in LTP and LTD, a reduction in neurogenesis, and in dendritic sprouting. The potential behavioural consequences of such impacts are discussed. It is hoped that this article will allow for greater examination of the effects of inflammatory factors on brain function, most particularly in terms of their role in chronic conditions.

An Examination of the Experiences of Practitioners Delivering Sport Psychology Services within English Premier League Soccer Academies.

Sport psychology has become increasingly recognized and accepted within professional sports, including soccer. To date, there is a lack of research that examines the provision of sport psychology within elite soccer, particularly from the experience of applied practitioners working within the field. The current study adopted a qualitative, inductive approach, to examine the experiences of practitioners responsible for sport psychology delivery within elite soccer academies in England. Seven participants (four females; three males), working within academies in the English Premier League, took part in semi-structured interviews about their experience of delivering sport psychology services within elite soccer academies. Results demonstrated that the provision of sport psychology is continually evolving, yet there are a number of factors that appear to inhibit the full integration of the discipline into academy soccer. Six key themes were identified: The breadth of sport psychology provision; what is sport psychology; the stigma surrounding sport psychology services; psychological literacy; the elite youth soccer environment; and the delivery of sport psychology under the Elite Player Performance Plan. Participants identified a lack of psychological literacy among coaches and academy staff, as well as a low level of guidance regarding the provision of psychology within the England Football Association's guiding document-the Elite Player Performance Plan-leading to considerable variation in the nature of the sport psychology provision. Future research would do well to also sample from a range of staff working within English soccer academies, in order to assess their perception of the level of provision and understanding of psychology.

Gender-Based Violence Against Trans* Individuals: A Netnography of Mary Gregory's Experience in Powerlifting.

In the context of sport, a growing body of research has reported the prevalence of violence against athletes, including sexual, physical, and psychological violence and neglect, experienced by both women and men in sport. Preliminary research has reported that gender-diverse individuals, specifically transgender athletes, may have a greater vulnerability to experiences of violence in sport, but this remains an under-researched population. In addition to limited research specifically on violence experienced by transgender athletes in sport, there is also only emerging research on virtual violence against athletes, with previous research on virtual violence in sporting spaces highlighting how online spaces are sites that can foster widespread hostility and violence. This study builds on previous research by examining discourses of virtual violence faced by transgender powerlifter, Mary Gregory, following her expulsion from the 100% Raw Powerlifting Federation. This research used a netnographic approach-an online ethnographic case study design. Data were collected from online news sources, as well as social media platforms, including Instagram, Twitter, and YouTube and were analyzed using reflexive thematic analysis. The data provided an insight into the cyberculture of powerlifting, and the negotiation of space, or lack thereof, for Mary Gregory within this physical culture. Five themes of were generated, including invalidation of gender identity, dehumanization, infliction of derogatory and crude language, accusations of cheating, and being compared to cisgender athletes without nuance. The study highlights the presence of significant vitriol across virtual platforms directed at Mary Gregory and the underlying presence of negative gender-based violence again trans* (GBV-T*) discourse. This case provides examples of virtual gender-based violence and transphobia in sport, a lack of readiness to accept trans* athletes, and concerns for the safety of trans* athletes in sporting spaces.

N-Linked glycosylation profiles of therapeutic induced senescent (TIS) triple negative breast cancer cells (TNBC) and their extracellular vesicle (EV) progeny.

Triple negative breast cancer (TNBC) has poor clinical outcomes and limited treatment options. Chemotherapy, while killing some cancer cells, can result in therapeutic-induced-senescent (TIS) cells. Senescent cells release significantly more extracellular vesicles (EVs) than non-senescent cells. Recently, N- and O-linked glycosylation alterations have been associated with senescence. We aimed to profile the N-linked glycans of whole cells, membrane, cytoplasm and EVs harvested from TIS TNBC cells and to compare these to results from non-senescent cells. TIS was induced in the Cal51 TNBC cells using the chemotherapeutic agent paclitaxel (PTX). Ultra-performance liquid chromatography (UPLC) analysis of exoglycosidase digested N-linked glycans was carried out on TIS compared to non-treated control cells. LC-Mass spectrometry (MS) analysis of the N-linked glycans and lectin blotting of samples was carried out to confirm the UPLC results. Significant differences were found in the N-glycan profile of the Cal51 membrane, cytoplasm and EV progeny of TIS compared to non-senescent cells. Protein mass spectrometry showed that the TIS cells contain different glycan modifying enzymes. The lectin, calnexin demonstrated a lower kDa size (∼58 kDa) in TIS compared to control cells (∼90 kDa) while Galectin 3 demonstrated potential proteolytic cleavage with 32 kDa and ∼22 kDa bands evident in TIS compared to non-senescent control cells with a major 32 kDa band only. TIS CAL51 cells also demonstrated a reduced adhesion to collagen I compared to control non-senescent cells. This study has shown that therapeutic-induced-senescent TNBC cells and their EV progeny, display differential N-glycan moieties compared to non-senescent Cal51 cells and their resultant EV progeny. For the future, N-glycan moieties on cancer senescent cells and their EV progeny hold potential for (i) the monitoring of treatment response as a liquid biopsy, and (ii) cancer senescent cell targeting with lectin therapies.

Targeting histone deacetylase 3 (HDAC3) in the bone marrow microenvironment inhibits multiple myeloma proliferation by modulating exosomes and IL-6 trans-signaling.

Multiple myeloma (MM) is an incurable cancer that derives pro-survival/proliferative signals from the bone marrow (BM) niche. Novel agents targeting not only cancer cells, but also the BM-niche have shown the greatest activity in MM. Histone deacetylases (HDACs) are therapeutic targets in MM and we previously showed that HDAC3 inhibition decreases MM proliferation both alone and in co-culture with bone marrow stromal cells (BMSC). In this study, we investigate the effects of HDAC3 targeting in BMSCs. Using both BMSC lines as well as patient-derived BMSCs, we show that HDAC3 expression in BMSCs can be induced by co-culture with MM cells. Knock-out (KO), knock-down (KD), and pharmacologic inhibition of HDAC3 in BMSCs results in decreased MM cell proliferation; including in autologous cultures of patient MM cells with BMSCs. We identified both quantitative and qualitative changes in exosomes and exosomal miRNA, as well as inhibition of IL-6 trans-signaling, as molecular mechanisms mediating anti-MM activity. Furthermore, we show that HDAC3-KD in BM endothelial cells decreases neoangiogenesis, consistent with a broad effect of HDAC3 targeting in the BM-niche. Our results therefore support the clinical development of HDAC3 inhibitors based not only on their direct anti-MM effects, but also their modulation of the BM microenvironment.

Validating a Self-Report Measure of Student Athletes' Perceived Stress Reactivity: Associations With Heart-Rate Variability and Stress Appraisals.

Student athletes experience multiple stressors relating to both their sporting and academic commitments. Individual differences play a significant role in how well student athletes cope with the demands they face. When assessing individual differences in stress reactivity, there are a lack of valid alternatives to costly and time-consuming lab-based physiological methods (e.g., cortisol sampling, cardiac variables). This paper aims to further validate a self-report measure of adolescent athletes' individual differences in perceived stress reactivity, by comparing to a psycho-physiological measure of emotion regulation (heart-rate variability) assessed during a socially evaluated cold pressor test. 30 student athletes and 31 student non-athletes completed a measure of perceived stress reactivity and took part in the socially evaluated cold pressor test while their heart-rate variability was assessed, along with their self-reported appraisals of stress, pain, and unpleasantness experienced during the procedure. Controlling for gender and athleticism, individual differences in perceived stress reactivity showed no associations with tonic or phasic levels of heart-rate variability. However, perceived stress reactivity was associated with levels of self-reported stress, pain, and unpleasantness experienced during the socially evaluated cold pressor test. These findings therefore suggest that perceived stress reactivity is associated with cognitive responses to stress (i.e., stress appraisals). However, further research is needed to confirm its relationship with physiological measures and responses. This further adds to the understanding of perceived stress reactivity, and validity of the perceived stress reactivity scale for adolescent athletes.

A Path Analysis of Adolescent Athletes' Perceived Stress Reactivity, Competition Appraisals, Emotions, Coping, and Performance Satisfaction.

This study examined a path analysis of adolescent athletes' individual differences in perceived stress reactivity, competition appraisals, emotions, coping, and performance satisfaction. The study aimed to extend an analysis by Nicholls et al. (2012) and further validate the use of the Perceived Stress Reactivity Scale for Adolescent Athletes (PSRS-AA). Adolescent athletes (N = 229, M age = 18.55, SD = 2.40) completed the PSRS-AA followed by a measure of competition appraisals less than 1 h before a competitive event. Within an hour after the competitive event, participants completed a retrospective assessment of emotions, coping strategies, and subjective performance. A path analysis revealed that perceived stress reactivity had direct and indirect effects on the appraisal of higher stressor intensity, lower perceived control, higher perceived threat, negative emotions, and maladaptive coping. Increased threat, positive and negative emotions, and maladaptive coping were associated with performance satisfaction. However, task-orientated coping was not associated with performance satisfaction. The present study enhances and refines the validity of the PSRS-AA for assessing adolescent athletes' perceived stress reactivity. Further strengths and weaknesses of the present study are discussed, along with recommendations for practitioners aiming to support adolescent athletes with high levels of stress reactivity.

An Exploration of the Mechanism of Action of an Equine-Assisted Intervention.

Though long alluded to, there is now an accumulation of evidence of the vital contribution that emotion makes to learning. Within this broad advance in understanding is a growing body of research emphasising the embodied nature of this emotion-based learning. The study presented here is a pilot study using a mixed-method approach (combining both physiological and experiential methodologies) to give a picture of the "emotional landscape" of people's learning through the intervention under study. This has allowed researchers to examine mediating pathways that may underlie any effects of an equine-assisted intervention. This study specifically focuses on examining the role of emotion. The intervention under study was used with young people with chronic mental health and behavioural problems for whom talk-based interventions were not working. Nine healthy participants aged 18-24 undertook the equine intervention, with an initial group having emotion-related psycho-physiological changes (skin conductance responses) measured while viewing their experience on video, and a further two participants experiencing a development of the methodology as their physiological responses were captured in real time during the intervention. The sessions were analysed by a group of five cross-disciplinary researchers to determine when significant learning episodes occurred, and the findings were that this learning was associated with powerful skin conductance responses. The qualitative element of the research entailed the participants watching themselves on video undertaking the equine intervention. They were asked to stop the video and share any changes in emotion at any point while watching. All participants experienced a positive temporal change in mood as the intervention progressed. All results supported the findings that emotional arousal occurred in relation to the participants asking the horse to perform a task. This paper will offer two novel contributions: (1) description of a new methodology for investigating the mechanism of action occurring in this type of intervention and (2) findings from the exploration of the intervention via psycho-physiological and experiential mechanisms.

Targeting Proteotoxic Stress in Cancer: A Review of the Role that Protein Quality Control Pathways Play in Oncogenesis.

Despite significant advances in cancer diagnostics and therapeutics the majority of cancer unfortunately remains incurable, which has led to continued research to better understand its exceptionally diverse biology. As a result of genomic instability, cancer cells typically have elevated proteotoxic stress. Recent appreciation of this functional link between the two secondary hallmarks of cancer: aneuploidy (oxidative stress) and proteotoxic stress, has therefore led to the development of new anticancer therapies targeting this emerging "Achilles heel" of malignancy. This review highlights the importance of managing proteotoxic stress for cancer cell survival and provides an overview of the integral role proteostasis pathways play in the maintenance of protein homeostasis. We further review the efforts undertaken to exploit proteotoxic stress in multiple myeloma (as an example of a hematologic malignancy) and triple negative breast cancer (as an example of a solid tumor), and give examples of: (1) FDA-approved therapies in routine clinical use; and (2) promising therapies currently in clinical trials. Finally, we provide new insights gleaned from the use of emerging technologies to disrupt the protein secretory pathway and repurpose E3 ligases to achieve targeted protein degradation.

Coping related variables, cardiac vagal activity and working memory performance under pressure.

The aim of this study was to assess the predictive role of coping related variables (trait emotional intelligence and reinvestment, challenge and threat appraisals and cardiac vagal activity) on cardiac vagal activity and working memory under low pressure (LP) and high pressure (HP) conditions. Participants (n = 49) completed trait questionnaires, the Decision Specific Reinvestment Scale, the Movement Specific Reinvestment Scale and Trait Emotional Intelligence Questionnaire. They realized the automated span task, which tests working memory, under counterbalanced LP and HP conditions. Cardiac vagal activity measurements were taken at rest, task and post task for 5 min, along with self-reported ratings of stress. Upon completion of the task, self-report measures of motivation, stress appraisal, attention and perceived pressure were completed. Current findings suggest cardiac vagal activity at rest can predict cardiac vagal activity under pressure, decision reinvestment influences cardiac vagal activity in cognitive tasks under LP and working memory performance is predicted by task cardiac vagal activity in HP only. These results show the importance of combining both subjective and objective psychophysiological variables in performance prediction and strengthen the need for this approach to be adopted across samples.

Resident bacteria in breast cancer tissue: pathogenic agents or harmless commensals?

Breast cancer is the second most common cancer in women. Recent evidence identifies a unique microbiome in breast tissue; a site previously thought to be sterile. The identification that this microbiome varies considerably from healthy subjects to cancer patients has prompted investigations into the role of specific bacterial species in oncogenesis. Indeed, certain bacteria have been shown to aid cancer development in vitro by promoting genomic instability, invasion, and chemotherapy resistance. However, the in vivo role of the breast microbiome in cancer appears to be more complex, involving numerous interactions between its constituent species and host cells. As such, reduced abundances of species which exert a protective effect against oncogenesis have come into focus and there is an emerging consensus that states of microbial dysbiosis, in which the normal balance of bacterial species is altered, can contribute to the development of cancer. This review summarizes the findings to date from the available literature pertaining to the microbiome in breast cancer and outlines areas worthy of further investigation.

The contribution of coping related variables and cardiac vagal activity on the performance of a dart throwing task under pressure.

The aims of this study were 1) to assess the predictive role of coping related variables (CRV) on cardiac vagal activity (derived from heart rate variability), and 2) to investigate the influence of CRV (including cardiac vagal activity) on a dart throwing task under low pressure (LP) and high pressure (HP) conditions. Participants (n=51) completed trait CRV questionnaires: Decision Specific Reinvestment Scale, Movement Specific Reinvestment Scale and Trait Emotional Intelligence Questionnaire. They competed in a dart throwing task under LP and HP conditions. Cardiac vagal activity measurements were taken at resting, task and during recovery for 5min. Self-reported ratings of stress were recorded at three time points via a visual analogue scale. Upon completion of the task, self-report measures of motivation, stress appraisal, attention, perceived pressure and dart throwing experience were completed. Results indicated that resting cardiac vagal activity had no predictors. Task cardiac vagal activity was predicted by resting cardiac vagal activity in both pressure conditions with the addition of a trait CRV in HP. Post task cardiac vagal activity was predicted by resting cardiac vagal activity in both conditions with the addition of a trait CRV in HP. Cardiac vagal reactivity (difference from resting to task) was predicted by a trait CRV in HP conditions. Cardiac vagal recovery (difference from task to post task) was predicted by a state CRV only in LP. Dart throwing task performance was predicted by a combination of both CRV and cardiac vagal activity. The current research suggests that coping related variables and cardiac vagal activity influence dart throwing task performance differently dependent on pressure condition.

Keratan sulfate epitopes exhibit a conserved distribution during joint development that remains undisclosed on the basis of glycosaminoglycan charge density.

Changes in glycosaminoglycan (GAG) content and distribution are vital for joint development. However, their precise character has not been established. We have used immunohistochemistry (IHC) and "critical electrolyte" Alcian blue staining to assess such changes in developing chick and rabbit joints. IHC showed chondroitin sulfate labeling in chick epiphyseal cartilage but not in interzones. In contrast, prominent labeling for keratan sulfate (KS) was restricted to chick cartilage-interzone interfaces. In rabbit knees, KS labeling was also prominent at presumptive cavity borders, but weak in interzone and cartilage. Selective pre-digestion produced appropriate loss of label and undersulfated KS was undetectable. Quantification of Alcian blue staining by scanning and integrating microdensitometry showed prominent hyaluronan-like (HA-like) interzone staining, with chondroitin sulfate and weaker KS staining restricted to epiphyseal cartilage. Hyaluronidase decreased HA-like staining in the interzone. Surprisingly, keratanases also reduced HA-like but not sulfated GAG (sGAG-like) staining in the interzone. Chondroitinase ABC had little effect on HA-like staining but decreased sGAG staining in all regions. Rabbit joints also showed HA-like but not KS staining in the interzone and strong chondroitin sulfate-like staining in epiphyseal cartilage. Our findings show restricted KS distribution in the region close to the presumptive joint cavity of developing chick and rabbit joints. Alcian blue staining does not detect this moiety. Therefore, it appears that although histochemistry allows relatively insensitive quantitative assessment of GAGs, IHC increases these detection limits. This is particularly evident for KS, which exhibits immunolabeling patterns in joints from different species that is consistent with a conserved functional role in chondrogenesis.

Microbial biotransformation of aryl sulfanylpentafluorides.

We report, for the first time, the biotransformation of potential pollutants bearing the pentafluorosulfanyl (SF5-) functional group in a fungus and bacteria. Cunninghamella elegans transformed p-methoxy phenyl SF5 via demethylation; Pseudomonas knackmussii and P. pseudoalcaligenes KF707 transformed amino-, hydroxyamino- and diamino- substituted phenyl SF5, forming the N-acetylated derivatives as the main product. Cell-free extract of Streptomyces griseus transformed 4-amino-3-hydroxy-phenyl SF5 to the N-acetylated derivative in the presence of acetyl CoA, confirming that an N-acetyltransferase is responsible for the bacterial biotransformations. Approximately 25% of drugs and 30% of agrochemicals contain fluorine, and the trifluoromethyl group is a prominent feature of many of these since it improves lipophilicity and stability. The pentafluorosulfanyl substituent is seen as an improvement on the trifluoromethyl group and research efforts are underway to develop synthetic methods to incorporate this moiety into biologically active compounds. It is important to determine the potential environmental impact of these compounds, including the potential biotransformation reactions that may occur when they are exposed to microorganisms.

Affirmation through disability: one athlete's personal journey to the London Paralympic Games.

AIMS: This article explores the personal narrative of a British Paralympic wheelchair tennis player who experienced a spinal cord injury (SCI) following a motorcycle accident in 2001 that left her paralysed from the waist down. The study responds to the call by Swain and French, among others, for alternative accounts of disability that demonstrate how life following impairment need not be empty and meaningless, but can actually reflect a positive, if different, social identity. METHODS: This study draws upon life history data to investigate the journey of one athlete who has managed to achieve international sporting success following a life-changing accident. A pseudonym has not been used for this study as the athlete wanted to be named in the research account and for her story to be shared. RESULTS: A chronological approach was adopted to map the pre- and post-accident recovery process. The account examines life before the trauma, the impact of the accident, the process of rehabilitation and the journey to athletic accomplishment. CONCLUSIONS: Negative views of disability can be challenged if disability is viewed in the context of positive life narratives. The story of one Paralympian demonstrates how an 'ordinary' person has made the most of an extraordinary situation and become a world-class athlete. This paper demonstrates that in contrast to typical discourse in disability studies, becoming disabled or living with a disability need not be a tragedy but may on the contrary enhance life and lead to positive affirmation.