CD4−CD8− T cells control intracellular bacterial infections both in vitro and in vivo

Memory T cells, including the well-known CD4+ and CD8+ T cells, are central components of the acquired immune system and are the basis for successful vaccination. After infection, CD4+ and CD8+ T cells expand into effector cells, and then differentiate into long-lived memory cells. We show that a rare population of CD4−CD8−CD3+αβ+γδ−NK1.1− T cells has similar functions. These cells potently and specifically inhibit the growth of the intracellular bacteria Mycobacterium tuberculosis (M. tb.) or Francisella tularensis Live Vaccine Strain (LVS) in macrophages in vitro, promote survival of mice infected with these organisms in vivo, and adoptively transfer immunity to F. tularensis LVS. Furthermore, these cells expand in the spleens of mice infected with M. tb. or F. tularensis LVS, and then acquire a memory cell phenotype. Thus, CD4−CD8− T cells have a role in the control of intracellular infection and may contribute to successful vaccination

Heart Rate and Energy Expenditure Concurrent Validity of Identical Garmin Wrist Watches During Moderately Heavy Resistance Training

Consistent with previous years, ACSM has found that wearable technology and resistance training (RT) are two of the top 5 fitness trends in 2023. Our lab recently found that wrist-worn devices, such as Garmin Instinct, are neither valid nor reliable at measuring average or maximal heart rate (HR) or estimating energy expenditure (EE) following light intensity circuit RT. We postulated that the errors may have been due to the device’s algorithms assuming higher intensity during RT. PURPOSE: The purpose of this study was to determine the concurrent validity of identical Garmin Instinct wrist-watches to record valid measures of average and maximal HR as well as estimated EE following moderately heavy RT. METHODS: Twenty-one adult participants completed this study (n=10 female, n=11 male). Two Garmin Instinct wrist-watches were evaluated, along with the Polar H10 chest strap and Cosmed K5 portable metabolic unit as the criterion devices for average/maximal HR and EE, respectively. Participants completed 8 supersets of the reverse lunge and shoulder press exercises using dumbbells at a light (4 sets) and moderately heavy (4 sets) intensity with 1 superset of 6 repetitions per exercise (12 repetitions per superset) and 1 min rest between supersets. Data were analyzed for validity (Mean Absolute Percent Error [MAPE] and Lin’s Concordance Coefficient [CCC]), with predetermined thresholds of MAPE\u3c10% and CCC\u3e0.70. A one-way repeated measures ANOVA with Sidak post-hoc test was used to determine differences (p\u3c0.05). RESULTS: The identical Garmin Instinct devices were not considered valid for average HR (MAPE range: 36.5-81.6%; CCC range: 0.07-0.18), maximal HR (MAPE range: 18.6-18.8%; CCC range: 0.15-0.31), or estimated EE (MAPE range: 14.0-16.4%; CCC range: 0.08-0.32) compared to the criterion references. The devices were significantly different than each other for average HR (p=0.005), maximal HR (p\u3c0.001), and estimated EE (p\u3c0.0001). CONCLUSION: The wearable wrist-worn devices tested herein should not be utilized for accurate measurements of HR or EE during RT, and there are even differences between identical devices. People who RT while using these devices should do so with caution if wishing to utilize them for physiological measures

Rating of Perceived Exertion, Average Heart Rate, and Energy Expenditure Following Indoor and Outdoor Moderately Heavy Superset Resistance Training

Our lab recently found that light intensity circuit resistance training outdoors had a significantly lower perception of effort (RPE) compared to indoor resistance training, despite no physiological differences in heart rate and energy expenditure. However, no study has examined other intensities or set schemes in differing environmental settings. PURPOSE: To determine how indoor or outdoor environments effect rating of perceived exertion (RPE) following light and moderately heavy intensity superset resistance training in recreationally resistance trained adults. METHODS: Twenty-three adult participants completed this study (n=10 female, n=13 male; age: 26.1±8.8 yrs; height: 172.2±9.5 cm; mass: 73.4±18.7 kg; RT experience: 5.3±4.8 yrs). Participants wore devices to measure heart rate (Polar H10 chest strap) and energy expenditure (Cosmed K5 Portable Metabolic Cart). Randomly in indoor and outdoor settings, participants completed 4 supersets of the reverse lunge and shoulder press exercises using dumbbells at a light (2 sets) and moderately heavy (2 sets) intensity with 1 superset of 6 repetitions per exercise (12 repetitions per superset) and 1 min rest between supersets. The OMNI Rating of Perceived Exertion Scale for Resistance Exercise 0-10 RPE scale was used following each superset. A paired T-test was used to determine differences between environmental setting (pRESULTS: No significant differences were observed between indoor and outdoor environments for average heart rate (129.4±17.2 and 127.8±23.3 bpm, p=0.67), energy expenditure (30.6±11.5 and 28.3±9.9 kcals; p=0.06), as well as RPE during light intensity (2.9±0.9 and 2.9±0.8 arbitrary units/AU’s, p=0.70) and moderately heavy intensity (6.5±1.7 and 6.3±1.5 AU’s, p=0.27) supersets. CONCLUSION: In recreationally resistance trained adults, light intensity and moderately heavy intensity superset resistance training in indoor or outdoor settings does not alter heart rate, energy expenditure, or perceived effort

Distinct physiological and behavioural functions for parental alleles of imprinted Grb10

Imprinted genes, defined by their preferential expression of a single parental allele, represent a subset of the mammalian genome and often have key roles in embryonic development1, but also postnatal functions including energy homeostasis2 and behaviour3, 4. When the two parental alleles are unequally represented within a social group (when there is sex bias in dispersal and/or variance in reproductive success)5, 6, imprinted genes may evolve to modulate social behaviour, although so far no such instance is known. Predominantly expressed from the maternal allele during embryogenesis, Grb10 encodes an intracellular adaptor protein that can interact with several receptor tyrosine kinases and downstream signalling molecules7. Here we demonstrate that within the brain Grb10 is expressed from the paternal allele from fetal life into adulthood and that ablation of this expression engenders increased social dominance specifically among other aspects of social behaviour, a finding supported by the observed increase in allogrooming by paternal Grb10-deficient animals. Grb10 is, therefore, the first example of an imprinted gene that regulates social behaviour. It is also currently alone in exhibiting imprinted expression from each of the parental alleles in a tissue-specific manner, as loss of the peripherally expressed maternal allele leads to significant fetal and placental overgrowth. Thus Grb10 is, so far, a unique imprinted gene, able to influence distinct physiological processes, fetal growth and adult behaviour, owing to actions of the two parental alleles in different tissues

Efficacy and pharmacokinetic/pharmacodynamic evaluation of the Aurora kinase A inhibitor MLN8237 against preclinical models of pediatric cancer

To gain a greater understanding of the potential of the Aurora kinase A inhibitor MLN8237 in the treatment of pediatric malignancies. The activity of MLN8237 was evaluated against 28 neuroblastoma and Ewing sarcoma cell lines, and its in vivo efficacy was studied over a range of doses against 12 pediatric tumor xenograft models. Pharmacokinetic, pharmacodynamic, and genomic studies were undertaken. In vitro neuroblastoma cell lines were generally more sensitive to MLN8237 than Ewing sarcoma lines. MLN8237 demonstrated significant activity in vivo against solid tumor models at the maximum tolerated dose (MTD); however, only 2 of 6 neuroblastoma models had objective responses at 0.25MTD. In contrast, MLN8237 induced objective responses at its MTD and at 0.5MTD in three ALL models and in two out of three at 0.25MTD. Pharmacokinetic studies at 0.5MTD demonstrated a T (max) of 0.5 h, C (max) of 24.8 mu M, AUC((0-24)) of 60.3 mu M h, and 12 h trough level of 1.2 mu M. Mitotic indices increased 6-12 h after MLN8237 administration. AURKA copy number variation was frequent in xenografts, and expression was highly correlated with copy number. Objective responses were more frequent in tumors with decreased AURKA copy number (5/8) compared to those with increased gene copy number (2/14). This report confirms the significant activity against both solid tumor and ALL xenografts at the MTD, with a steep dose response. These data support clinical development of MLN8237 in childhood cancer. Because of the steep dose-response relationship, such studies should target achieving trough levels of 1 mu M or higher for sustained periods of treatment

Ghrelin Indirectly Activates Hypophysiotropic CRF Neurons in Rodents

Ghrelin is a stomach-derived hormone that regulates food intake and neuroendocrine function by acting on its receptor, GHSR (Growth Hormone Secretagogue Receptor). Recent evidence indicates that a key function of ghrelin is to signal stress to the brain. It has been suggested that one of the potential stress-related ghrelin targets is the CRF (Corticotropin-Releasing Factor)-producing neurons of the hypothalamic paraventricular nucleus, which secrete the CRF neuropeptide into the median eminence and activate the hypothalamic-pituitary-adrenal axis. However, the neural circuits that mediate the ghrelin-induced activation of this neuroendocrine axis are mostly uncharacterized. In the current study, we characterized in vivo the mechanism by which ghrelin activates the hypophysiotropic CRF neurons in mice. We found that peripheral or intra-cerebro-ventricular administration of ghrelin strongly activates c-fos – a marker of cellular activation – in CRF-producing neurons. Also, ghrelin activates CRF gene expression in the paraventricular nucleus of the hypothalamus and the hypothalamic-pituitary-adrenal axis at peripheral level. Ghrelin administration directly into the paraventricular nucleus of the hypothalamus also induces c-fos within the CRF-producing neurons and the hypothalamic-pituitary-adrenal axis, without any significant effect on the food intake. Interestingly, dual-label immunohistochemical analysis and ghrelin binding studies failed to show GHSR expression in CRF neurons. Thus, we conclude that ghrelin activates hypophysiotropic CRF neurons, albeit indirectly

Whole-genome sequencing reveals host factors underlying critical COVID-19

Critical COVID-19 is caused by immune-mediated inflammatory lung injury. Host genetic variation influences the development of illness requiring critical care1 or hospitalization2–4 after infection with SARS-CoV-2. The GenOMICC (Genetics of Mortality in Critical Care) study enables the comparison of genomes from individuals who are critically ill with those of population controls to find underlying disease mechanisms. Here we use whole-genome sequencing in 7,491 critically ill individuals compared with 48,400 controls to discover and replicate 23 independent variants that significantly predispose to critical COVID-19. We identify 16 new independent associations, including variants within genes that are involved in interferon signalling (IL10RB and PLSCR1), leucocyte differentiation (BCL11A) and blood-type antigen secretor status (FUT2). Using transcriptome-wide association and colocalization to infer the effect of gene expression on disease severity, we find evidence that implicates multiple genes—including reduced expression of a membrane flippase (ATP11A), and increased expression of a mucin (MUC1)—in critical disease. Mendelian randomization provides evidence in support of causal roles for myeloid cell adhesion molecules (SELE, ICAM5 and CD209) and the coagulation factor F8, all of which are potentially druggable targets. Our results are broadly consistent with a multi-component model of COVID-19 pathophysiology, in which at least two distinct mechanisms can predispose to life-threatening disease: failure to control viral replication; or an enhanced tendency towards pulmonary inflammation and intravascular coagulation. We show that comparison between cases of critical illness and population controls is highly efficient for the detection of therapeutically relevant mechanisms of disease

Genetic mechanisms of critical illness in COVID-19.

Host-mediated lung inflammation is present1, and drives mortality2, in the critical illness caused by coronavirus disease 2019 (COVID-19). Host genetic variants associated with critical illness may identify mechanistic targets for therapeutic development3. Here we report the results of the GenOMICC (Genetics Of Mortality In Critical Care) genome-wide association study in 2,244 critically ill patients with COVID-19 from 208 UK intensive care units. We have identified and replicated the following new genome-wide significant associations: on chromosome 12q24.13 (rs10735079, P = 1.65 × 10-8) in a gene cluster that encodes antiviral restriction enzyme activators (OAS1, OAS2 and OAS3); on chromosome 19p13.2 (rs74956615, P = 2.3 × 10-8) near the gene that encodes tyrosine kinase 2 (TYK2); on chromosome 19p13.3 (rs2109069, P = 3.98 ×  10-12) within the gene that encodes dipeptidyl peptidase 9 (DPP9); and on chromosome 21q22.1 (rs2236757, P = 4.99 × 10-8) in the interferon receptor gene IFNAR2. We identified potential targets for repurposing of licensed medications: using Mendelian randomization, we found evidence that low expression of IFNAR2, or high expression of TYK2, are associated with life-threatening disease; and transcriptome-wide association in lung tissue revealed that high expression of the monocyte-macrophage chemotactic receptor CCR2 is associated with severe COVID-19. Our results identify robust genetic signals relating to key host antiviral defence mechanisms and mediators of inflammatory organ damage in COVID-19. Both mechanisms may be amenable to targeted treatment with existing drugs. However, large-scale randomized clinical trials will be essential before any change to clinical practice

Metabolic and Functional Genomic Studies Identify Deoxythymidylate Kinase as a Target in LKB1-Mutant Lung Cancer

The LKB1/STK11 tumor suppressor encodes a serine/threonine kinase which coordinates cell growth, polarity, motility, and metabolism. In non-small cell lung cancer, LKB1 is somatically inactivated in 25-30% of cases, often concurrently with activating KRAS mutation. Here, we employed an integrative approach to define novel therapeutic targets in KRAS-driven LKB1 mutant lung cancers. High-throughput RNAi screens in lung cancer cell lines from genetically engineered mouse models driven by activated KRAS with or without coincident Lkb1 deletion led to the identification of Dtymk, encoding deoxythymidylate kinase which catalyzes dTTP biosynthesis, as synthetically lethal with Lkb1 deficiency in mouse and human lung cancer lines. Global metabolite profiling demonstrated that Lkb1-null cells had striking decreases in multiple nucleotide metabolites as compared to the Lkb1-wt cells. Thus, LKB1 mutant lung cancers have deficits in nucleotide metabolism conferring hypersensitivity to DTYMK inhibition, suggesting that DTYMK is a potential therapeutic target in this aggressive subset of tumors