Receptor binding and immunogenic properties of the receptor binding domain of influenza D virus hemagglutinin-esterase-fusion protein expressed from Escherichia coli.

The hemagglutinin-esterase-fusion (HEF) protein binds 9-O-acetylated sialic acids-containing glycans on the cell surface and drives influenza D virus (IDV) entry. The HEF is a primary determinant of the exceptional thermal and acid stability observed in IDV infection biology. Here, we expressed and purified the receptor binding domain (RBD) of the IDV HEF protein in Escherichia coli and characterized its receptor binding and antigenic properties. The data from these experiments indicate that (i) the RBD can bind with specificity to turkey red blood cells (RBC), and its binding can be specifically inhibited by IDV antibody; (ii) the RBD efficiently binds to the cell surface of MDCK cells expressing the receptor of IDV; and (iii) anti-RBD antibodies are capable of blocking RBD attachment to MDCK cells as well as of inhibiting the virus from agglutinating RBCs. These observations support the utility of this RBD in future receptor and entry studies of IDV.

The Characteristics of Endurance Events with a Variable Pacing Profile-Time to Embrace the Concept of "Intermittent Endurance Events"?

A variable pacing profile is common in different endurance events. In these races, several factors, such as changes in elevation or race dynamics, lead participants to perform numerous surges in intensity. These surges are so frequent that certain events, such as cross-country (XC) skiing, mountain biking (MTB), triathlon, and road cycling, have been termed "intermittent endurance events". The characteristics of these surges vary depending on the sport: MTB and triathlon require athletes to perform numerous short (<10 s) bouts; XC skiing require periods of short- and moderate-(30 s to 2 min) duration efforts, while road cycling is comprised of a mix of short-, moderate-, and long-duration (>2 min) bouts. These bouts occur at intensities above the maximal metabolic steady state (MMSS), with many efforts performed at intensities above the athletes' maximal aerobic power or speed (MAP/MAS) (i.e., supramaximal intensities). Given the factors that influence the requirement to perform surges in these events, athletes must be prepared to always engage in a race with a highly stochastic pace. The aim of this review is to characterize the variable pacing profile seen in endurance events and to discuss how the performance of multiple maximal and supramaximal surges in intensity can affect how athletes fatigue during a race and influence training strategies that can lead to success in these races.

Real-World Analysis of the Clinical and Economic Impact of the 21-Gene Recurrence Score (RS) in Invasive Lobular Early-Stage Breast Carcinoma in Ireland.

Background: This study, using real-world data, assesses the impact of RS testing on treatment pathways and the associated economic consequences of such testing. This paper pertains to lobular breast cancer. Methods: A retrospective, observational study was undertaken between 2011 and 2019 on a cross-section of hormone receptor-positive (HR+), HER2-negative, lymph node-negative, early-stage breast cancer patients. All patients had ILC and had RS testing in Ireland. The patient population is representative of the national population. Patients were classified as low (RS ≤ 25) or high (RS > 25) risk. Patients aged ≤50 were stratified as low (RS 0-15), intermediate (RS 16-25), or high risk (RS > 25). Results: A total of 168 patients were included, most of whom had grade 2 (G2) tumors (n = 154, 92%). Overall, 155 patients (92.3%) had low RS (≤25), 12 (7.1%) had high RS (>25), and 1 (0.6%) had unknown RS status. In 29 (17.5%) patients aged ≤50 at diagnosis, RS was ≤15 in 16 (55%), 16-20 in 6 (21%), 21-25 in 5 (17%), >25 in 1 (3.5%), and unknown in 1 (3.5%). Post RS testing, 126 patients (78%) had a change in chemotherapy recommendation; all to hormone therapy. In total, only 35 patients (22%) received chemotherapy. RS testing achieved a 75% reduction in chemotherapy use, resulting in savings of €921,543.84 in treatment costs, and net savings of €387,283.84. Conclusions: The use of this test resulted in a 75% reduction in chemotherapy and a significant cost savings in our publicly funded health system.

Influence of an Exercise-Specific Face Mask on Physiological and Perceptual Responses to Graded Exercise.

The impact of exercise-specific face masks (ESFMs) in aerobically fit individuals on physiological, perceptual, respiratory, and performance responses remains unclear. How ESFMs mitigate exercise-induced bronchoconstriction (EIB) is also unknown. Thus, this study aimed to determine how an ESFM altered within-exercise physiological, perceptual, respiratory, and performance responses to graded treadmill exercise. Twenty-four individuals (11 females) completed a discontinuous graded exercise test on a treadmill under two conditions (ESFM and unmasked). Physiological, respiratory function, and perceptual measures were assessed. Performance was determined by time to exhaustion. Statistical analyses included linear mixed-effects modeling, repeated measures analysis of variance, and pairwise comparisons using an alpha value of 0.05. ESFM use significantly impaired performance (median = -150.5 s) and decreased arterial oxygen saturation at maximal intensity (mean = -3.7%). Perceptions of air hunger and work of breathing were elevated across submaximal and maximal intensities. Perceived exertion and breathing discomfort were significantly elevated submaximally but not maximally. Spirometry measures were not significantly different at termination but were significantly improved at submaximal intensities in participants with and without EIB. ESFM use in fit individuals increased perceptual discomfort, impaired performance, and augmented arterial desaturation. Respiratory function improvements were observed but were accompanied by adverse perceptual sensations. Despite this, performance impairments may limit the real-world utility of ESFMs for athletes.

Repeated Short-Term Bouts of Hyperoxia Improve Aerobic Performance in Acute Hypoxia.

ABSTRACT: Faulhaber, M, Schneider, S, Rausch, LK, Dünnwald, T, Menz, V, Gatterer, H, Kennedy, MD, and Schobersberger, W. Repeated short-term bouts of hyperoxia improve aerobic performance in acute hypoxia. J Strength Cond Res 37(10): 2016-2022, 2023-This study aimed to test the effects of repeated short-term bouts of hyperoxia on maximal 5-minute cycling performance under acute hypoxic conditions (3,200 m). Seventeen healthy and recreationally trained individuals (7 women and 10 men) participated in this randomized placebo-controlled cross-over trial. The procedures included a maximal cycle ergometer test and 3 maximal 5-minute cycling time trials (TTs). TT1 took place in normoxia and served for habituation and reference. TT2 and TT3 were conducted in normobaric hypoxia (15.0% inspiratory fraction of oxygen). During TT2 and TT3, the subjects were breathing through a face mask during five 15-second periods. The face mask was connected through a nonrebreathing T valve to a 300-L bag filled with 100% oxygen (intermittent hyperoxia) or ambient hypoxic air (placebo). The main outcome was the mean power output during the TT. Statistical significance level was set at p < 0.05. The mean power output was higher in the intermittent hyperoxia compared with the placebo condition (255.5 ± 49.6 W vs. 247.4 ± 48.2 W, p = 0.001). Blood lactate concentration and ratings of perceived exertion were significantly lower by about 9.7 and 7.3%, respectively, in the intermittent hyperoxia compared with the placebo condition, whereas heart rate values were unchanged. IH application increased arterial oxygen saturation (82.9 ± 2.6% to 92.4 ± 3.3%, p < 0.001). Repeated 15-second bouts of hyperoxia, applied during high-intensity exercise in hypoxia, are sufficient to increase power output. Future studies should focus on potential dose-response effects and the involved mechanisms.

Association of personal and equipment-related factors on ACL injury risk in alpine skiers with cautious or risk-taking behaviour: A case-control study.

Background: In recreational alpine skiing ACL injury risk depends on the interaction of individual characteristics and behaviours as well as on equipment-related factors. Aim: to evaluate if and to what extent personal characteristics and equipment-related parameters are associated with ACL injury risk in cautious and risk-taking recreational alpine skiers. Methods: A retrospective questionnaire-based, case-control study of ACL-injured and uninjured in a cohort of cautious as well as risk-taking recreational skiers was conducted. Participants self-reported their demographics, skiing skill level, and risk-taking behaviour. Ski length, side-cut radius, widths of the tip, waist, and tail were recorded from each participant's skis. Standing heights at the front and rear components of the ski binding were measured with a digital sliding caliper, and a standing height ratio between the front and rear components was calculated. Ski boot sole abrasion at the toe and heel pieces was also measured with the digital sliding caliper. Results: In total, 1068 recreational skiers (50.8% females) with a mean age of 37.8 ± 12.3 years participated, of whom 193 (22.0%) sustained an ACL injury, and 330 (30.9%) participants reported a risk-taking behaviour. Results of the multiple logistic regression analyses revealed that a higher age, a lower skill level, a higher standing height ratio, and greater ski boot sole abrasion at the toe as well as heel pieces were independently associated with an increased ACL injury risk in both the cautious and the risk-taking group. Among cautious skiers, a longer ski length was an additional significant risk factor for sustaining an ACL injury. In conclusion, the same personal and equipment related characteristics contribute to an increase in the ACL injury risk regardless of risk-taking behaviour, with the only difference that longer skis represent an additional risk factor in cautious skiers.

Statistical considerations and database limitations in NMR-based metabolic profiling studies.

INTRODUCTION: Interpretation and analysis of NMR-based metabolic profiling studies is limited by substantially incomplete commercial and academic databases. Statistical significance tests, including p-values, VIP scores, AUC values and FC values, can be largely inconsistent. Data normalization prior to statistical analysis can cause erroneous outcomes. OBJECTIVES: The objectives were (1) to quantitatively assess consistency among p-values, VIP scores, AUC values and FC values in representative NMR-based metabolic profiling datasets, (2) to assess how data normalization can impact statistical significance outcomes, (3) to determine resonance peak assignment completion potential using commonly used databases and (4) to analyze intersection and uniqueness of metabolite space in these databases. METHODS: P-values, VIP scores, AUC values and FC values, and their dependence on data normalization, were determined in orthotopic mouse model of pancreatic cancer and two human pancreatic cancer cell lines. Completeness of resonance assignments were evaluated using Chenomx, the human metabolite database (HMDB) and the COLMAR database. The intersection and uniqueness of the databases was quantified. RESULTS: P-values and AUC values were strongly correlated compared to VIP or FC values. Distributions of statistically significant bins depended strongly on whether or not datasets were normalized. 40-45% of peaks had either no or ambiguous database matches. 9-22% of metabolites were unique to each database. CONCLUSIONS: Lack of consistency in statistical analyses of metabolomics data can lead to misleading or inconsistent interpretation. Data normalization can have large effects on statistical analysis and should be justified. About 40% of peak assignments remain ambiguous or impossible with current databases. 1D and 2D databases should be made consistent to maximize metabolite assignment confidence and validation.

Blind assessment of monomeric AlphaFold2 protein structure models with experimental NMR data.

Recent advances in molecular modeling of protein structures are changing the field of structural biology. AlphaFold-2 (AF2), an AI system developed by DeepMind, Inc., utilizes attention-based deep learning to predict models of protein structures with high accuracy relative to structures determined by X-ray crystallography and cryo-electron microscopy (cryoEM). Comparing AF2 models to structures determined using solution NMR data, both high similarities and distinct differences have been observed. Since AF2 was trained on X-ray crystal and cryoEM structures, we assessed how accurately AF2 can model small, monomeric, solution protein NMR structures which (i) were not used in the AF2 training data set, and (ii) did not have homologous structures in the Protein Data Bank at the time of AF2 training. We identified nine open-source protein NMR data sets for such "blind" targets, including chemical shift, raw NMR FID data, NOESY peak lists, and (for 1 case) 15N-1H residual dipolar coupling data. For these nine small (70-108 residues) monomeric proteins, we generated AF2 prediction models and assessed how well these models fit to these experimental NMR data, using several well-established NMR structure validation tools. In most of these cases, the AF2 models fit the NMR data nearly as well, or sometimes better than, the corresponding NMR structure models previously deposited in the Protein Data Bank. These results provide benchmark NMR data for assessing new NMR data analysis and protein structure prediction methods. They also document the potential for using AF2 as a guiding tool in protein NMR data analysis, and more generally for hypothesis generation in structural biology research.

Creatine supplementation and endurance performance: surges and sprints to win the race.

Creatine supplementation is an effective ergogenic aid to augment resistance training and improve intense, short duration, intermittent performance. The effects on endurance performance are less known. The purpose of this brief narrative review is to discuss the potential mechanisms of how creatine can affect endurance performance, defined as large muscle mass activities that are cyclical in nature and are >~3 min in duration, and to highlight specific nuances within the literature. Mechanistically, creatine supplementation elevates skeletal muscle phosphocreatine (PCr) stores facilitating a greater capacity to rapidly resynthesize ATP and buffer hydrogen ion accumulation. When co-ingested with carbohydrates, creatine enhances glycogen resynthesis and content, an important fuel to support high-intensity aerobic exercise. In addition, creatine lowers inflammation and oxidative stress and has the potential to increase mitochondrial biogenesis. In contrast, creatine supplementation increases body mass, which may offset the potential positive effects, particularly in weight-bearing activities. Overall, creatine supplementation increases time to exhaustion during high-intensity endurance activities, likely due to increasing anaerobic work capacity. In terms of time trial performances, results are mixed; however, creatine supplementation appears to be more effective at improving performances that require multiple surges in intensity and/or during end spurts, which are often key race-defining moments. Given creatines ability to enhance anaerobic work capacity and performance through repeated surges in intensity, creatine supplementation may be beneficial for sports, such as cross-country skiing, mountain biking, cycling, triathlon, and for short-duration events where end-spurts are critical for performance, such as rowing, kayaking, and track cycling.

Do Female University Varsity Athletes Have a Greater Risk of Injury Within a Competitive Varsity Season?

Previous varsity sport injury research has analyzed how acute and chronic injury severity, type, and location differs between sport and sexes, with limited research in time to injury. Canadian university varsity sport injury research is especially sparse and mostly retrospective. Thus, we aimed to understand injury differences in male and female competitive university athletes competing in the same sport. Athletes who competed on the basketball, volleyball, soccer, ice hockey, football (male), rugby (female), and wrestling teams were eligible for the study. There were 182 male and 113 female athletes who provided informed consent to be prospectively followed over a season. Injury date, type, location, chronicity, and events missed due to injury were recorded on a weekly basis. Overall, the percentage of male (68.7%) and female (68.1%) athletes injured was not different. No overall sex differences (variables collapsed) were observed in injury chronicity, location, type, events lost, mean number of injuries, or time to injury. Within sport differences existed for mean number of injuries, injury location, type of injury, and events missed. Mean time to injury in female basketball (28 days) and volleyball athletes (14 days) was significantly shorter compared to male basketball (67 days) and volleyball (65 days). Time to a concussion was significantly shorter in females overall compared to males. These results indicate that Canadian female university age athletes are not inherently more susceptible to injury, but female athletes within certain sports may have increased injury risk which could shorten time to injury (basketball, volleyball) and increase the number of events missed due to injury (hockey).

Effect of incline versus block heel-raise exercise on foot muscle strength and vertical jump performance - an 11-week randomized resistance training study.

Strengthening the toe flexors and ankle plantar flexors may improve vertical jump performance. One exercise that may be effective for concurrently strengthening these muscles is heel-raises performed on an incline. The purpose of this study was to investigate the effects of incline versus conventional (block) heel-raise exercise on hallux and II-V digit flexor strength, vertical jump performance, and ankle plantar flexor strength. Thirty-three female volleyball players were randomly allocated to perform incline (n = 17) or block (n = 16) heel-raise exercise for 11-weeks. Participants' toe flexor strength, countermovement jump, approach jump, and ankle plantar flexor strength were assessed before, after 7 weeks, and after 11 weeks of exercise. There were no significant time-by-group interactions for any variable (p > 0.05). However, both groups improved their hallux flexor strength (Δ0.27 ± 0.50 N·kg-1; p < 0.05), and vertical countermovement (Δ1.2 ± 2.3 cm; p < 0.05) and approach (Δ1.9 ± 2.6 cm; p < 0.05) jump height from pre- to post-test. No changes were observed in the ankle plantar flexor or II-V digit flexor strength (n > 0.05). Both incline and conventional heel-raises improve toe flexor strength. Practitioners seeking to improve individuals' foot function may consider incorporating incline or block heel-raises.

Safety and efficacy of autologous cell vaccines in solid tumors: a systematic review and meta-analysis of randomized control trials.

We conducted a systematic review and meta-analysis of randomized control trials to formally assess the safety and efficacy of autologous whole cell vaccines as immunotherapies for solid tumors. Our primary safety outcome was number, and grade of adverse events. Our primary efficacy outcome was clinical responses. Secondary outcomes included survival metrics and correlative immune assays. We searched MEDLINE, Embase, and the Cochrane Central Register of Controlled Trials for studies published between 1946 and August 2020 using any autologous whole cell product in the treatment of any solid tumor. The Cochrane Randomized Controlled Trial risk of bias tool was used to assess risk of bias. Eighteen manuscripts were identified with a total of 714 patients enrolled in control and 808 in vaccine arms. In 698 patients receiving at least one dose of vaccine, treatment was well tolerated with a total of 5 grade III or higher adverse events. Clinical response was reported in a minority (n = 2, 14%) of studies. Autologous cell vaccines were associated with improved overall (HR 1.28, 95% CI 1.01-1.63) and disease-free survival (HR 1.33, 95% CI 1.05-1.67) over thirteen and ten trials respectively. Where reported, immune assays correlated well with clinical outcomes. Our results suggest that autologous whole cell vaccination is safe and efficacious in increasing survival in patients undergoing treatment for solid tumors.Registration: PROSPERO CRD42019140187.

Blind Assessment of Monomeric AlphaFold2 Protein Structure Models with Experimental NMR Data.

Recent advances in molecular modeling of protein structures are changing the field of structural biology. AlphaFold-2 (AF2), an AI system developed by DeepMind, Inc., utilizes attention-based deep learning to predict models of protein structures with high accuracy relative to structures determined by X-ray crystallography and cryo-electron microscopy (cryoEM). Comparing AF2 models to structures determined using solution NMR data, both high similarities and distinct differences have been observed. Since AF2 was trained on X-ray crystal and cryoEM structures, we assessed how accurately AF2 can model small, monomeric, solution protein NMR structures which (i) were not used in the AF2 training data set, and (ii) did not have homologous structures in the Protein Data Bank at the time of AF2 training. We identified nine open source protein NMR data sets for such "blind" targets, including chemical shift, raw NMR FID data, NOESY peak lists, and (for 1 case) 15 N- 1 H residual dipolar coupling data. For these nine small (70 - 108 residues) monomeric proteins, we generated AF2 prediction models and assessed how well these models fit to these experimental NMR data, using several well-established NMR structure validation tools. In most of these cases, the AF2 models fit the NMR data nearly as well, or sometimes better than, the corresponding NMR structure models previously deposited in the Protein Data Bank. These results provide benchmark NMR data for assessing new NMR data analysis and protein structure prediction methods. They also document the potential for using AF2 as a guiding tool in protein NMR data analysis, and more generally for hypothesis generation in structural biology research. Highlights: AF2 models assessed against NMR data for 9 monomeric proteins not used in training.AF2 models fit NMR data almost as well as the experimentally-determined structures. RPF-DP, PSVS , and PDBStat software provide structure quality and RDC assessment. RPF-DP analysis using AF2 models suggests multiple conformational states.

Preventing Surgery-Induced NK Cell Dysfunction Using Anti-TGF-ß Immunotherapeutics.

Natural Killer (NK) cell cytotoxicity and interferon-gamma (IFNγ) production are profoundly suppressed postoperatively. This dysfunction is associated with increased morbidity and cancer recurrence. NK activity depends on the integration of activating and inhibitory signals, which may be modulated by transforming growth factor-beta (TGF-ß). We hypothesized that impaired postoperative NK cell IFNγ production is due to altered signaling pathways caused by postoperative TGF-ß. NK cell receptor expression, downstream phosphorylated targets, and IFNγ production were assessed using peripheral blood mononuclear cells (PBMCs) from patients undergoing cancer surgery. Healthy NK cells were incubated in the presence of healthy/baseline/postoperative day (POD) 1 plasma and in the presence/absence of a TGF-ß-blocking monoclonal antibody (mAb) or the small molecule inhibitor (smi) SB525334. Single-cell RNA sequencing (scRNA-seq) was performed on PBMCs from six patients with colorectal cancer having surgery at baseline/on POD1. Intracellular IFNγ, activating receptors (CD132, CD212, NKG2D, DNAM-1), and downstream target (STAT5, STAT4, p38 MAPK, S6) phosphorylation were significantly reduced on POD1. Furthermore, this dysfunction was phenocopied in healthy NK cells through incubation with rTGF-ß1 or POD1 plasma and was prevented by the addition of anti-TGF-ß immunotherapeutics (anti-TGF-ß mAb or TGF-ßR smi). Targeted gene analysis revealed significant decreases in S6 and FKBP12, an increase in Shp-2, and a reduction in NK metabolism-associated transcripts on POD1. pSmad2/3 was increased and pS6 was reduced in response to rTGF-ß1 on POD1, changes that were prevented by anti-TGF-ß immunotherapeutics. Together, these results suggest that both canonical and mTOR pathways downstream of TGF-ß mediate phenotypic changes that result in postoperative NK cell dysfunction.

The Adequacy of Prenatal Care in Rural Kansas Related to Distance Traveled.

Introduction: Prenatal care is essential for optimizing the health of a woman and her baby. Multiple factors have created barriers in the access to prenatal care in rural Kansas. Over 120 rural hospitals in the United States have closed since 2010, 5 in Kansas. Seventy-seven of the 105 Kansas counties do not have maternity care services. This study investigated differences in prenatal care received by women in rural Kansas counties related to distance traveled. Differences in timing of initiation of care, number of visits, and services received were compared between two cohorts: those who drove < 19 miles and those who drove ≥ 20 miles for prenatal care. Methods: A survey was distributed to women who had delivered a child in the last three years in rural Kansas at participating clinics. Measures of adequacy of prenatal care were determined with questions regarding timing of first prenatal visit, number of prenatal visits, and services received at visits. An index was created using these variables and compared between the two cohorts using two-tailed t-tests for continuous data and chi square analysis for categorical data. Results: Women who traveled ≥ 20 miles for prenatal care received statistically significant less services, and had less prenatal care visits in the second trimester and overall in their pregnancy compared to women who traveled < 19 miles for prenatal care. Rurality did not impact adequacy of prenatal care. Conclusions: Women traveling ≥ 20 miles to receive prenatal care had significantly fewer prenatal visits during their second trimester and overall in pregnancy and self-reported less prenatal care services. These results indicated the importance of lessening barriers to prenatal care in rural Kansas, such as transportation and financial barriers.

Birth Outcomes Related to Distance in Rural and Frontier Kansas.

Introduction: Women from rural communities must travel greater distances to secure obstetrical care. This study sought the extent to which distance traveled by mothers for obstetrical services affects birth outcomes in rural and frontier counties of Kansas. Methods: Medical students invited women over the age of 18 to participate in a recall survey regarding their children under three years old. Participants were a sample of convenience, and the length of data collection was a month. A bivariate analysis was performed on the responses gathered regarding obstetrical measures as a function of self-reported distance traveled to the hospital of delivery. Results: Eighty-five women completed the survey, but only 76 satisfied all eligibility requirements. No statistical difference in birth outcomes were found between women who travel more than or less than 20 miles. However, when correlating data to that of the Kansas Hospital Association and the Kansas Department of Health and Environment, counties without birth facilities had a higher percentage of very low birth weights (< 1,500 grams) and more babies born at full-term when compared to counties that offer birth facilities. Babies born to mothers who reside in counties with obstetrical services were born at an earlier gestational age than those without birth facilities. Lastly, babies born into a family with income less than $50,000 weighed less and had a shorter gestational age than those from a more affluent household. Conclusions: The results revealed counterintuitive findings that deserve to be explored further by a study with greater statistical power.

A Translational Randomized Trial of Perioperative Arginine Immunonutrition on Natural Killer Cell Function in Colorectal Cancer Surgery Patients.

BACKGROUND: Surgery results in severe impairment of natural killer (NK) cell cytotoxicity (NKC) and activity (NKA, cytokine secretion), and a dramatic drop in arginine levels. Postoperative immunosuppression is associated with increased complications and recurrence. Perioperative arginine is reported to reduce postoperative complications. Because arginine modulates NK cell function, this study aimed to determine whether perioperative consumption of arginine-enriched supplements (AES) can improve NK cell function in colorectal cancer (CRC) surgery patients. METHODS: This study randomized 24 CRC patients to receive the AES or isocaloric/isonitrogenous control supplement three times a day for five days before and after surgery. The AES contained 4.2 g of arginine per dose (12.6 g/day). The primary objective was to determine whether AES improved NKC by 50 % compared with the control group after surgery. RESULTS: On surgery day (SD) 1, NKC was significantly reduced postoperatively in the control group by 50 % (interquartile range [IQR], 36-55 %; p = 0.02) but not in the AES group (25 % reduction; IQR, 28-75 %; p = 0.3). Furthermore, AES had no benefit in terms of NKA or NK cell number. Compliance was much greater preoperatively (>91 %) than postoperatively (<46 %). However, despite excellent preoperative compliance, arginine was rapidly cleared from the blood within 4 h after consumption and therefore, did not prevent the postoperative drop in arginine. CONCLUSIONS: Oral consumption of arginine immunonutrition resulted in a modest improvement in NKC after surgery but was unable to prevent postoperative arginine depletion or the suppression of NKA (ClinicalTrials.gov NCT02987296).

Inhaled seralutinib exhibits potent efficacy in models of pulmonary arterial hypertension.

BACKGROUND: Signalling through platelet-derived growth factor receptor (PDGFR), colony-stimulating factor 1 receptor (CSF1R) and mast/stem cell growth factor receptor kit (c-KIT) plays a critical role in pulmonary arterial hypertension (PAH). We examined the preclinical efficacy of inhaled seralutinib, a unique small-molecule PDGFR/CSF1R/c-KIT kinase inhibitor in clinical development for PAH, in comparison to a proof-of-concept kinase inhibitor, imatinib. METHODS: Seralutinib and imatinib potency and selectivity were compared. Inhaled seralutinib pharmacokinetics/pharmacodynamics were studied in healthy rats. Efficacy was evaluated in two rat models of PAH: SU5416/Hypoxia (SU5416/H) and monocrotaline pneumonectomy (MCTPN). Effects on inflammatory/cytokine signalling were examined. PDGFR, CSF1R and c-KIT immunohistochemistry in rat and human PAH lung samples and microRNA (miRNA) analysis in the SU5416/H model were performed. RESULTS: Seralutinib potently inhibited PDGFRα/ß, CSF1R and c-KIT. Inhaled seralutinib demonstrated dose-dependent inhibition of lung PDGFR and c-KIT signalling and increased bone morphogenetic protein receptor type 2 (BMPR2). Seralutinib improved cardiopulmonary haemodynamic parameters and reduced small pulmonary artery muscularisation and right ventricle hypertrophy in both models. In the SU5416/H model, seralutinib improved cardiopulmonary haemodynamic parameters, restored lung BMPR2 protein levels and decreased N-terminal pro-brain natriuretic peptide (NT-proBNP), more than imatinib. Quantitative immunohistochemistry in human lung PAH samples demonstrated increased PDGFR, CSF1R and c-KIT. miRNA analysis revealed candidates that could mediate seralutinib effects on BMPR2. CONCLUSIONS: Inhaled seralutinib was an effective treatment of severe PAH in two animal models, with improved cardiopulmonary haemodynamic parameters, a reduction in NT-proBNP, reverse remodelling of pulmonary vascular pathology and improvement in inflammatory biomarkers. Seralutinib showed greater efficacy compared to imatinib in a preclinical study.

Perioperative arginine prevents metastases by accelerating natural killer cell recovery after surgery.

Profound natural killer (NK) cell suppression after cancer surgery is a main driver of metastases and recurrence, for which there is no clinically approved intervention available. Surgical stress is known to cause systemic postoperative changes that negatively modulate NK cell function including the expansion of surgery-induced myeloid-derived suppressor cells (Sx-MDSCs) and a marked reduction in arginine bioavailability. In this study, we determine that Sx-MDSCs regulate systemic arginine levels in the postoperative period and that restoring arginine imbalance after surgery by dietary intake alone was sufficient to significantly reduce surgery-induced metastases in our preclinical murine models. Importantly, the effects of perioperative arginine were dependent upon NK cells. Although perioperative arginine did not prevent immediate NK cell immunoparalysis after surgery, it did accelerate their return to preoperative cytotoxicity, interferon gamma secretion, and activating receptor expression. Finally, in a cohort of patients with colorectal cancer, postoperative arginine levels were shown to correlate with their Sx-MDSC levels. Therefore, this study lends further support for the use of perioperative arginine supplementation by improving NK cell recovery after surgery.