Effect of high-fat diet on isometric, concentric and eccentric contractile performance of skeletal muscle isolated from female CD-1 mice.

Despite evidence inferring muscle and contractile mode-specific effects of high-fat diet (HFD), no study has yet considered the impact of HFD directly on eccentric muscle function. The present work uniquely examined the effect of 20-week HFD on the isometric, concentric and eccentric muscle function of isolated mouse soleus (SOL) and extensor digitorum longus (EDL) muscles. CD-1 female mice were randomly split into a control (n = 16) or HFD (n = 17) group and for 20 weeks consumed standard lab chow or HFD. Following this period, SOL and EDL muscles were isolated and assessments of maximal isometric force and concentric work loop (WL) power were performed. Each muscle was then subjected to either multiple concentric or eccentric WL activations. Post-fatigue recovery, as an indicator of incurred damage, was measured via assessment of concentric WL power. In the EDL, absolute concentric power and concentric power normalised to muscle mass were reduced in the HFD group (P < 0.038). HFD resulted in faster concentric fatigue and reduced eccentric activity-induced muscle damage (P < 0.05). For the SOL, maximal isometric force was increased, and maximal eccentric power normalised to muscle mass and concentric fatigue were reduced in the HFD group (P < 0.05). HFD effects on eccentric muscle function are muscle-specific and have little relationship with changes in isometric or concentric function. HFD has the potential to negatively affect the intrinsic concentric and eccentric power-producing capacity of skeletal muscle, but a lack of a within-muscle uniform response indicates disparate mechanisms of action which require further investigation.

Force-Time Characteristics of Repeated Bouts of Depth Jumps and the Effects of Compression Garments.

No studies have reported ground reaction force (GRF) profiles of the repeated depth jump (DJ) protocols commonly used to study exercise-induced muscle damage. Furthermore, while compression garments (CG) may accelerate recovery from exercise-induced muscle damage, any effects on the repeated bout effect are unknown. Therefore, we investigated the GRF profiles of 2 repeated bouts of damage-inducing DJs and the effects of wearing CG for recovery. Nonresistance-trained males randomly received CG (n = 9) or placebo (n = 8) for 72 hours recovery, following 20 × 20 m sprints and 10 × 10 DJs from 0.6 m. Exercise was repeated after 14 days. Using a 3-way (set × bout × group) design, changes in GRF were assessed with analysis of variance and statistical parametric mapping. Jump height, reactive strength, peak, and mean propulsive forces declined between sets (P < .001). Vertical stiffness, contact time, force at zero velocity, and propulsive duration increased (P < .05). According to statistical parametric mapping, braking (17%-25% of the movement) and propulsive forces (58%-81%) declined (P < .05). During the repeated bout, peak propulsive force and duration increased (P < .05), while mean propulsive force (P < .05) and GRF from 59% to 73% declined (P < .001). A repeated bout of DJs differed in propulsive GRF, without changes to the eccentric phase, or effects from CG.

Serum concentration impacts myosin heavy chain expression but not cellular respiration in human LHCN-M2 myoblasts undergoing differentiation.

NEW FINDINGS: What is the central question of this study? Does the concentration of human serum affect skeletal muscle differentiation and cellular respiration of LHCN-M2 myoblasts? What is the main finding and its importance? The concentration of serum used to differentiate LHCN-M2 skeletal muscle cells impacts the coverage of myosin heavy chain, a marker of terminally differentiated myotubes. Normalisation of mitochondrial function data to total protein negates the differences observed in absolute values, which differ as a result of increased protein content when differentiation occurs with increasing concentration of serum. ABSTRACT: The human LHCN-M2 myoblast cell line has the potential to be used to investigate skeletal muscle development and metabolism. Experiments were performed to determine how different concentrations of human serum affect myogenic differentiation and mitochondrial function of LHCN-M2 cells. LHCN-M2 myoblasts were differentiated in serum-free medium, 0.5% or 2% human serum for 5 and 10 days. Myotube formation was assessed by immunofluorescence staining of myosin heavy chain (MHC) and molecularly by mRNA expression of Myogenic differentiation 1 (MYOD1) and Myoregulatory factor 5 (MYF5). Following differentiation, mitochondrial function was assessed to establish the impact of serum concentration on mitochondrial function. Time in differentiation increased mRNA expression of MYOD1 (day 5, 6.58 ± 1.33-fold; and day 10, 4.28 ± 1.71-fold) (P = 0.012), while suppressing the expression of MYF5 (day 5, 0.21 ± 0.11-fold; and day 10, 0.06 ± 0.03-fold) (P = 0.001), regardless of the serum concentration. Higher serum concentrations increased MHC area (serum free, 11.92 ± 0.85%; 0.5%, 23.10 ± 5.82%; 2%, 43.94 ± 8.92%) (P = 0.001). Absolute basal respiration approached significance (P = 0.06) with significant differences in baseline oxygen consumption rate (P = 0.025) and proton leak (P = 0.006) when differentiated in 2% human serum, but these were not different between conditions when normalised to total protein. Our findings show that increasing concentrations of serum of LHCN-M2 skeletal muscle cells into multinucleated myotubes, but this does not affect relative mitochondrial function.

The effect of medical grade compression garments on the repeated-bout effect in non-resistance-trained men.

NEW FINDINGS: What is the central question of this study? What are the effects of compression garments on recovery from unaccustomed damaging exercise and subsequent protective adaptations? What is the main finding and its importance? Compression did not influence recovery, but was associated with blunted protective adaptations for isokinetic performance, which were completely absent at high velocities. Based on these findings, the use of compression garments for recovery would not be recommended following unaccustomed exercise, particularly if the maintenance of high-velocity performance following exercise-induced muscle damage is desirable. ABSTRACT: Whilst compression garments (CG) may enhance recovery from exercise-induced muscle damage (EIMD), many recovery strategies can attenuate adaptative responses. Therefore, the effects of CG on recovery from EIMD, and the rapid protective adaptations known as the repeated bout effect (RBE) were investigated. Thirty-four non-resistance-trained males (18-45 years) randomly received class II medical-grade CG or placebo for 72 h following eccentrically-focused lower-body exercise, in a double-blind, randomised controlled trial. Indices of EIMD were assessed at baseline, 0, 24, 48 and 72 h post-exercise, before exercise and testing were repeated after 14 days. Results were analysed using a three-way (time × condition × bout) linear mixed-effects model. Exercise impaired isometric and isokinetic strength, with soreness and thigh circumference elevated for 72 h (P < 0.001). Compression did not enhance recovery (P > 0.05), despite small to moderate effect sizes (ES, reported alongside 90% confidence intervals) for isokinetic strength (ES from 0.2 [-0.41, 0.82] to 0.65 [0.03, 1.28]). All variables recovered faster after the repeated bout (P < 0.005). However, RBE for peak isokinetic force was impaired in CG at 60° s-1 (group × bout interaction: χ2  = 4.24, P = 0.0395; ES = -0.56 [-1.18, 0.07]) and completely absent at 120° s-1 (χ2  = 16.2, P < 0.001, ES = -0.96 [-1.61, -0.32]) and 180° s-1 (χ2  = 10.4, P = 0.001, ES = -0.72 [-1.35, -0.09]). Compression blunted RBE at higher isokinetic velocities without improving recovery in non-resistance-trained males, potentially contraindicating their use following unaccustomed exercise in this population.

Extreme occupational heat exposure is associated with elevated haematological and inflammatory markers in Fire Service Instructors.

NEW FINDINGS: What is the central question of this study? Fire service instructors are frequently exposed to live fire scenarios, representing the most extreme chronic occupational heat exposure. These individuals report a series of unique health issues. We sought to identify whether the number of exposures completed was associated with inflammatory and immunological markers and symptoms of ill health. What is the main finding and its importance? Fire service instructors exhibit greater levels of inflammatory markers in comparison to firefighters. The number of exposures to fire is positively related to the prevalence of ill health and inflammation. Implementation of a proposed limit of nine exposures per month might be appropriate to minimize health issues. ABSTRACT: Fire Service Instructors (FSIs) experience ∼10 times more fire exposures than firefighters (FFs), and the increased physiological stress from this potentially puts them at risk of ill health and future cardiac events. The aim of the study was to establish whether FSIs exhibit elevated biomarkers associated with cardiac event risk, identify whether FSIs experience systemic inflammation linked to the frequency of fire exposure and evaluate a proposed exposure limit of nine exposures per month. Blood samples were collected from 110 Fire Service personnel (mean ± SD, age,44 ± 7 years; height, 178.1 ± 7.1 cm; and body mass, 84.3 ± 12.0 kg; FSIs n = 53 and FFs n = 57) for biomarker analysis. Work history details were collected from all participants. Participants with biomarker concentrations above healthy reference ranges were classified as being 'at risk'. The neutrophil-to-lymphocyte ratio, platelet count, cardiac troponin T, interleukin (IL)-6, IL-1ß, C-reactive protein and immunoglobulin G were greater in FSIs than in FFs (P < 0.05). Multiple regression analysis revealed that 18.8% of IL-6, 24.9% of IL-1ß, 29.2% of C-reactive protein and 10.9% of immunoglobulin G variance could be explained by the number of exposures to heat per month. Odds ratios revealed that those FSIs above the nine per month exposure limit were six to 12 times more likely to be classified as 'at risk' and were 16 times more likely to experience symptoms of ill health. Increased cytokine levels suggest that FSIs experience systemic inflammation, which is related to symptoms of ill health. We propose that an exposure limit could reduce the prevalence of these biomarker risk factors and ill health.

A double-blind, placebo-controlled, randomised-designed GABA tea study in children diagnosed with autism spectrum conditions: a feasibility study clinical trial registration: ISRCTN 72571312.

Objective: The research has shown an association with sensorimotor integration and symptomology of Autism Spectrum Conditions (ASC). Specific areas of the brain that are involved in sensorimotor integration, such as the cerebellum and basal ganglia, are pathologically different in individuals with ASC in comparison to typically developing (TD) peers. These brain regions contain GABAergic inhibitory neurons that release an inhibitory neurotransmitter, γ-Aminobutyric acid (GABA). Brain GABA levels are decreased in ASC. This study explored the effect of introducing a non-invasive GABA substitute, in the form of GABA Oolong tea, on sensorimotor skills, ASC profiles, anxieties and sleep of children with ASC. Methods: Nine children took part: (5 male, 4 female). Each child participated in three tea conditions: high GABA, high L-Theanine (a compound that increases GABA), placebo with low GABA. A double-blind, repeated measures design was employed. Measures were taken after each tea condition. Sensory and ASC profiles were scored using parental questionnaires. Motor skills were assessed using a gold standard coordination assessment. Sleep was monitored using an actiwatch and anxiety measured through cortisol assays. Subjective views were sought from parents on 'best' tea. Results: The results showed significant improvement in manual dexterity and some large individual improvements in balance, sensory responsivity, DSM-5 criteria and cortisol levels with GABA tea. Improvements were also seen in the L-Theanine condition although they were more sporadic. Conclusions: These results suggest that sensorimotor abilities, anxiety levels and DSM-5 symptomology of children with ASC can benefit from the administration of GABA in the form of Oolong tea.

Acute Hormonal Response to Kettlebell Swing Exercise Differs Depending on Load, Even When Total Work Is Normalized.

ABSTRACT: Raymond, LM, Renshaw, D, and Duncan, MJ. Acute hormonal response to kettlebell swing exercise differs depending on load, even when total work is normalized. J Strength Cond Res 35(4): 997-1005, 2021-This study examined the acute hormonal response to kettlebell (KB) swing exercise using 2 loads, but when total work was equalized. Ten strength-trained males (25 ± 6 years) completed 2 KB swing trials, with an 8- and 16-kg KB, respectively, in a counterbalanced order. Each protocol lasted 12 minutes comprising 30-second KB swings followed by 30-second rest. Swing cadence was manipulated in each trial to ensure that total weight lifted was the same across conditions. Heart rate (HR) and ratings of perceived exertion (RPE), using the Borg RPE scale 6-20, were taken at the end of each 30-second exercise period. Saliva samples (min 0.5 ml) were taken 15 minutes before, immediately after, and 15 and 30 minutes after each condition from which cortisol (C) and testosterone (T) were determined. Results indicated a significant main effect for load for C (p = 0.007) and T (p = 0.05) where higher values for both C and T were evident for the 16-kg load. There was also a significant main effect for time for T (p = 0.001), where T values were all significantly higher post-exercise compared with pre-exercise. For HR, there were significant main effects for load (p = 0.004) and time (p = 0.001) with higher HR seen in 16-kg load and significant increases in HR evident with increasing repetition, irrespective of condition (all p < 0.05). Rating of perceived exertion values increased with repetition for the 8-kg and 16-kg loads, but the increase was more marked for the 16-kg load compared with the 8-kg load (p = 0.002). The present findings suggest that KB swing exercise produces an acute increase in hormones involved in muscle adaptation, but that KB load influences this response, even when total work completed is the same.

Integrin-associated ILK and PINCH1 protein content are reduced in skeletal muscle of maintenance haemodialysis patients.

KEY POINTS: Patients with renal failure undergoing maintenance haemodialysis are associated with insulin resistance and protein metabolism dysfunction. Novel research suggests that disruption to the transmembrane protein linkage between the cytoskeleton and the extracellular matrix in skeletal muscle may contribute to reduced amino acid metabolism and insulin resistance in haemodialysis. ILK, PINCH1 and pFAKTyr397 were significantly decreased in haemodialysis compared to controls, whereas Rac1 and Akt2 showed no different between groups. Rac1 deletion in the Rac1 knockout model did not alter the expression of integrin-associated proteins. Phenylalanine kinetics were reduced in the haemodialysis group at 30 and 60 min post meal ingestion compared to controls; both groups showed similar levels of insulin sensitivity and ß-cell function. Key proteins in the integrin-cytoskeleton linkage are reduced in haemodialysis patients, suggesting for the first time that integrin-associated proteins dysfunction may contribute to reduced phenylalanine flux without affecting insulin resistance in haemodialysis patients. ABSTRACT: Muscle atrophy, insulin resistance and reduced muscle phosphoinositide 3-kinase-Akt signalling are common characteristics of patients undergoing maintenance haemodialysis (MHD). Disruption to the transmembrane protein linkage between the cytoskeleton and the extracellular matrix in skeletal muscle may contribute to reduced amino acid metabolism and insulin resistance in MHD patients. Eight MHD patients (age: 56 ± 5 years: body mass index: 32 ± 2 kg m-2 ) and non-diseased controls (age: 50 ± 2 years: body mass index: 31 ± 1 kg m-2 ) received primed continuous l-[ring-2 H5 ]phenylalanine before consuming a mixed meal. Phenylalanine metabolism was determined using two-compartment modelling. Muscle biopsies were collected prior to the meal and at 300 min postprandially. In a separate experiment, skeletal muscle tissue from muscle-specific Rac1 knockout (Rac1 mKO) was harvested to investigate whether Rac1 depletion disrupted the cytoskeleton-integrin linkage, allowing for cross-model examination of proteins of interest. ILK, PINCH1 and pFAKTyr397 were significantly lower in MHD (P < 0.01). Rac1 and Akt showed no difference between groups for the human trial. Rac1 deletion in the Rac1 mKO model did not alter the expression of integrin-associated proteins. Phenylalanine rates of appearance and disappearance, as well as metabolic clearance rates, were lower in the MHD group at 30 and 60 min post meal ingestion compared to controls (P < 0.05). Both groups showed similar levels of insulin sensitivity and ß-cell function. Key proteins in the integrin-cytoskeleton linkage are reduced in MHD patients, suggesting for the first time that integrin-associated proteins dysfunction may contribute to reduced phenylalanine flux without affecting insulin resistance in haemodialysis patients.

Age-related degeneration of lumbar muscle morphology in healthy younger versus older men.

AIM: The aim of this study was to evaluate age-related changes in lumbar paravertebral muscle (LPM) morphology in healthy younger and older adult men. METHODS: T2-weighted axial MRI of the lumbar spine were obtained for 12 healthy older (67.3 ± 6.0 years) and younger (24.7 ± 3.1 years) men. Normalised muscle volume (NMV) and muscle fat infiltrate (MFI) were determined bilaterally for the psoas (PS), quadratus lumborum (QL), erector spinae (ES) and multifidus (MF). MANOVA was used to compare NMV and MFI between age groups. Follow-up ANOVA compared NMV and MFI for each muscle between age groups, with physical activity (PA) as a covariate. Stepwise regression was used to explore the association between muscle morphology. RESULTS: NMV of the ES and QL were significantly lower in the older group (OG) (p = 0.040 and p < 0.001, respectively). MFI across all muscles was significantly greater in the OG (p < 0.001). PA did not moderate the relationship between aging and muscle degeneration. Non-dominant handgrip strength was associated with NMV (p = 0.003). CONCLUSIONS: Age-related atrophy is muscle specific in the lumbar spine; changes in lumbar musculature is independent of PA, handgrip strength may reflect morphological changes in the postural muscles with age. This study supports establishing effective targeted exercise interventions in the lumbar musculature.

Tiotropium bromide, a long acting muscarinic receptor antagonist triggers intracellular calcium signalling in the heart.

BACKGROUND AND PURPOSE: Tiotropium bromide (TB) is a long acting muscarinic receptor antagonist used to manage chronic obstructive pulmonary disease (COPD). Recent meta-analyses suggest an increased risk of cardiovascular events with TB. Ca2+/calmodulin dependent kinase II (CaMKII) and L-type Ca2+ channels regulate Ca2+ concentrations allowing management of Ca2+ across membranes. Pathological increases in Ca2+ are initially slow and progressive, however once the cytosolic concentration rises >1-3 µM from ~100 nM, calcium overload occurs and can lead to cell death. Ipratropium bromide, a short acting muscarinic receptor antagonist has previously been found to induce Ca2+ mediated eryptosis. The aim of this study was to investigate the role of Ca2+ in Tiotropium bromide mediated cardiotoxicity. EXPERIMENTAL APPROACH: Isolated Sprague-Dawley rat hearts were perfused with TB (10-0.1 nM) ±â€¯KN-93 (400 nM) or nifedipine (1 nM). Hearts were stained to determine infarct size (%) using triphenyltetrazolium chloride (TTC), or snap frozen to determine p-CaMKII (Thr286) expression. Cardiomyocytes were isolated using a modified Langendorff perfusion and enzymatic dissociation before preparation for Fluo 3-AM staining and flow cytometric analysis. KEY RESULTS: TB increased infarct size compared to controls by 6.91-8.41%, with no effect on haemodynamic function. KN-93/nifedipine with TB showed a 5.90/7.38% decrease in infarct size compared to TB alone, the combined use of KN-93 with TB also showed a significant increase in left ventricular developed pressure whilst nifedipine with TB showed a significant decrease in coronary flow. TB showed a 42.73% increase in p-CaMKII (Thr286) versus control, and increased Ca2+ fluorescence by 30.63% in cardiomyocytes. CONCLUSIONS AND IMPLICATIONS: To our knowledge, this is the first pre-clinical study to show that Tiotropium bromide induces Ca2+ signalling via CaMKII and L-type Ca2+ channels to result in cell damage. This has significant clinical impact due to long term use of TB in COPD patients, and warrants assessment of cardiac drug safety.

Predictors of satisfaction and quality of life following post-mastectomy breast reconstruction.

OBJECTIVE: Breast reconstruction is associated with multiple psychological benefits. However, few studies have identified clinical and psychological factors associated with improved satisfaction and quality of life. This study examined factors, which predict satisfaction with breast appearance, outcome satisfaction and quality of life following post-mastectomy breast reconstruction. METHODS: Women who underwent post-mastectomy breast reconstruction between 2010 and 2016 received a postal questionnaire consisting of The BREAST-Q Patient Reported Outcomes Instrument, The European Organisation for Research and Treatment of Cancer QLQ-30 Questionnaire, The Patient and Observer Scar Assessment Scale, and a series of Visual-Analogue Scales. One hundredforty-eight women completed the questionnaire, a 56% response rate. RESULTS: Hierarchical multiple regression analyses revealed psychosocial factors accounted for 75% of the variance in breast satisfaction, 68% for outcome satisfaction, and 46% forquality of life. Psychosocial well-being emerged as a significant predictor of satisfaction with breast appearance (ß = .322) and outcome satisfaction (ß = .406). Deep inferior epigastric perforator flap patients reported greater satisfaction with breast appearance (ß = .120) and outcome satisfaction (ß = .167). CONCLUSIONS: This study extends beyond the limited research by distinguishing between satisfaction with breast appearance and outcome satisfaction. The study provides evidence for the role of psychosocial factors predicting key patient reported outcomes and demonstrates the importance of psychosocial well-being and reconstruction type. The findings also highlight the need for healthcare providers to consider the psychosocial well-being of patients both preoperatively and post operatively and provide preliminary evidence for the use of deep inferior epigastric perforator reconstructions over other types of reconstructive procedures.

Characterization of hepcidin response to holotransferrin in novel recombinant TfR1 HepG2 cells.

Hepcidin is the key regulator of systemic iron homeostasis. The iron-sensing mechanisms and the role of intracellular iron in modulating hepatic hepcidin secretion are unclear. Therefore, we created a novel cell line, recombinant-TfR1 HepG2, expressing iron-response-element-independent TFRC mRNA to promote cellular iron-overload and examined the effect of excess holotransferrin (5g/L) on cell-surface TfR1, iron content, hepcidin secretion and mRNA expressions of TFRC, HAMP, SLC40A1, HFE and TFR2. Results showed that the recombinant cells exceeded levels of cell-surface TfR1 in wild-type cells under basal (2.8-fold; p<0.03) and holotransferrin-supplemented conditions for 24h and 48h (4.4- and 7.5-fold, respectively; p<0.01). Also, these cells showed higher intracellular iron content than wild-type cells under basal (3-fold; p<0.03) and holotransferrin-supplemented conditions (6.6-fold at 4h; p<0.01). However, hepcidin secretion was not higher than wild-type cells. Moreover, holotransferrin treatment to recombinant cells did not elevate HAMP responses compared to untreated or wild-type cells. In conclusion, increased intracellular iron content in recombinant cells did not increase hepcidin responses compared to wild-type cells, resembling hemochromatosis. Furthermore, TFR2 expression altered within 4h of treatment, while HFE expression altered later at 24h and 48h, suggesting that TFR2 may function prior to HFE in HAMP regulation.

Risks associated with endotoxins in feed additives produced by fermentation.

Increasingly, feed additives for livestock, such as amino acids and vitamins, are being produced by Gram-negative bacteria, particularly Escherichia coli. The potential therefore exists for animals, consumers and workers to be exposed to possibly harmful amounts of endotoxin from these products. The aim of this review was to assess the extent of the risk from endotoxins in feed additives and to calculate how such risk can be assessed from the properties of the additive. Livestock are frequently exposed to a relatively high content of endotoxin in the diet: no additional hazard to livestock would be anticipated if the endotoxin concentration of the feed additive falls in the same range as feedstuffs. Consumer exposure will be unaffected by the consumption of food derived from animals receiving endotoxin-containing feed, because the small concentrations of endotoxin absorbed do not accumulate in edible tissues. In contrast, workers processing a dusty additive may be exposed to hazardous amounts of endotoxin even if the endotoxin concentration of the product is low. A calculation method is proposed to compare the potential risk to the worker, based on the dusting potential, the endotoxin concentration and technical guidance of the European Food Safety Authority, with national exposure limits.

Characterisation of hepcidin response to holotransferrin treatment in CHO TRVb-1 cells.

Iron overload coupled with low hepcidin levels are characteristics of hereditary haemochromatosis. To understand the role of transferrin receptor (TFR) and intracellular iron in hepcidin secretion, Chinese hamster ovary transferrin receptor variant (CHO TRVb-1) cells were used that express iron-response-element-depleted human TFRC mRNA (TFRC∆IRE). Results showed that CHO TRVb-1 cells expressed higher basal levels of cell-surface TFR1 than HepG2 cells (2.2-fold; p < 0.01) and following 5 g/L holotransferrin treatment maintained constitutive over-expression at 24h and 48 h, contrasting the HepG2 cells where the receptor levels significantly declined. Despite this, the intracellular iron content was neither higher than HepG2 cells nor increased over time under basal or holotransferrin-treated conditions. Interestingly, hepcidin secretion in CHO TRVb-1 cells exceeded basal levels at all time-points (p < 0.02) and matched levels in HepG2 cells following treatment. While TFRC mRNA expression showed expected elevation (2h, p < 0.03; 4h; p < 0.05), slc40a1 mRNA expression was also elevated (2 h, p < 0.05; 4 h, p < 0.03), unlike the HepG2 cells. In conclusion, the CHO TRVb-1 cells prevented cellular iron-overload by elevating slc40a1 expression, thereby highlighting its significance in the absence of iron-regulated TFRC mRNA. Furthermore, hepcidin response to holotransferrin treatment was similar to HepG2 cells and resembled the human physiological response.

Attenuation of plasma annexin A1 in human obesity.

Obesity-related metabolic disorders are characterized by mild chronic inflammation, leukocyte infiltration, and tissue fibrosis as a result of adipocytokine production from the expanding white adipose tissue. Annexin A1 (AnxA1) is an endogenous glucocorticoid regulated protein, which modulates systemic anti-inflammatory processes and, therefore, may be altered with increasing adiposity in humans. Paradoxically, we found that plasma AnxA1 concentrations inversely correlated with BMI, total percentage body fat, and waist-to-hip ratio in human subjects. Plasma AnxA1 was also inversely correlated with plasma concentrations of the acute-phase protein, C-reactive protein (CRP), and the adipocytokine leptin, suggesting that as systemic inflammation increases, anti-inflammatory AnxA1 is reduced. In addition, AnxA1 gene expression and protein were significantly up-regulated during adipogenesis in a human adipocyte cell line compared to vehicle alone, demonstrating for the first time that AnxA1 is expressed and excreted from human adipocytes. These data demonstrate a failure in the endogenous anti-inflammatory system to respond to increasing systemic inflammation resulting from expanding adipose tissue, a condition strongly linked to the development of type 2 diabetes and cardiovascular disease. These data raise the possibility that a reduction in plasma AnxA1 may contribute to the chronic inflammatory phenotype observed in human obesity.

Design and development of novel mitochondrial targeted nanocarriers, DQAsomes for curcumin inhalation.

Curcumin has potent antioxidant and anti-inflammatory properties but poor absorption following oral administration owing to its low aqueous solubility. Development of novel formulations to improve its in vivo efficacy is therefore challenging. In this study, formulation of curcumin-loaded DQAsomes (vesicles formed from the amphiphile, dequalinium) for pulmonary delivery is presented for the first time. The vesicles demonstrated mean hydrodynamic diameters between 170 and 200 nm, with a ζ potential of approximately +50 mV, high drug loading (up to 61%) and encapsulation efficiency (90%), resulting in enhanced curcumin aqueous solubility. Curcumin encapsulation in DQAsomes in the amorphous state was confirmed by X-ray diffraction and differential scanning calorimetry analysis. The existence of hydrogen bonds and cation-π interaction between curcumin and vesicle building blocks, namely dequalinium molecules, were shown in lyophilized DQAsomes using FT-IR analysis. Encapsulation of curcumin in DQAsomes enhanced the antioxidant activity of curcumin compared to free curcumin. DQAsome dispersion was successfully nebulized with the majority of the delivered dose deposited in the second stage of the twin-stage impinger. The vesicles showed potential for mitochondrial targeting. Curcumin-loaded DQAsomes thus represent a promising inhalation formulation with improved stability characteristics and mitochondrial targeting ability, indicating a novel approach for efficient curcumin delivery for effective treatment of acute lung injury and the rationale for future in vivo studies.

Formulated Palmitoylethanolamide Supplementation Improves Parameters of Cognitive Function and BDNF Levels in Young, Healthy Adults: A Randomised Cross-Over Trial.

BACKGROUND: Palmitoylethanolamide (PEA) is an endocannabinoid-like lipid mediator which is naturally produced in the body and found in certain foods. The aim of this study was to assess the effect of a bioavailable formulated form of PEA (Levagen+®) on serum BDNF levels and parameters of cognitive function in healthy adults. METHODS: A randomised double-blinded placebo-controlled cross-over trial was implemented to measure the effects of a 6-week 700 mg/day course of formulated PEA supplementation versus a placebo. Participants (n = 39) completed pre- and post-assessments of a lab-based cognitive test. Serum samples were collected to measure BDNF concentrations using an immunoassay. RESULTS: A significant increase in serum BDNF levels was found following PEA supplementation compared with the placebo (p = 0. 0057, d = 0.62). The cognition test battery demonstrated improved memory with PEA supplementation through better first success (p = 0.142, d = 0.54) and fewer errors (p = 0.0287; d = -0.47) on the Paired Associates Learning test. CONCLUSION: This was the first study to report a direct beneficial effect of Levagen+® PEA supplementation on memory improvement as well as corresponding increases in circulating neurotrophic marker levels. This suggests that formulated PEA holds promise as an innovative and practical intervention for cognitive health enhancement.

Effects of resistance exercise and whey protein supplementation on cognitive function in older men: secondary analysis of a randomised, double-blind, placebo-controlled trial.

PURPOSE: Ageing is associated with cognitive decline. This study investigated the individual and combined effects of resistance exercise (RE) and whey protein supplementation (PRO) on cognitive function in older men. METHODS: In a pooled-groups analysis, 36 older men (age: 67 ± 4 years) were randomised to either RE (2 x/week; n = 18) or no exercise (NE; n = 18), and either PRO (2 × 25 g/d whey protein isolate; n = 18) or control (CON, 2 × 23.75 g maltodextrin/d; n = 18). A sub-analysis was also conducted between RE + CON (n = 9) and RE + PRO (n = 9). At baseline and 12 weeks, participants completed a battery of neuropsychological tests (CANTAB; Cambridge Cognition, UK) and neurobiological, inflammatory, salivary cortisol and insulin sensitivity biomarkers were quantified. RESULTS: PRO improved executive function z-score (+0.31 ± 0.08) greater than CON (+0.06 ± 0.08, P = 0.03) and there was a trend towards improved global cognitive function (P = 0.053). RE and RE + PRO did not improve any cognitive function domains (p ≥ 0.07). RE decreased tumor necrosis factor-alpha (P = 0.02) and interleukin-6 (P = 0.048) concentrations compared to NE, but changes in biomarkers did not correlate with changes in cognitive domains. Muscle strength (r = 0.34, P = 0.045) and physical function (ρ = 0.35-0.51, P < 0.05) outcomes positively correlated with cognitive function domains at baseline, but only Δskeletal muscle index correlated with Δepisodic memory (r = 0.34, P = 0.046) following the intervention. CONCLUSION: In older men, PRO improved cognitive function, most notably executive functioning. RE did not improve any cognitive function domains but did decrease biomarkers of systemic inflammation. No synergistic effects were observed.

CellShip: An Ambient Temperature Transport and Short-Term Storage Medium for Mammalian Cell Cultures.

Cell culture is a critical platform for numerous research and industrial processes. However, methods for transporting cells are largely limited to cryopreservation, which is logistically challenging, requires the use of potentially cytotoxic cryopreservatives, and can result in poor cell recovery. Development of a transport media that can be used at ambient temperatures would alleviate these issues. In this study, we describe a novel transportation medium for mammalian cells. Five commonly used cell lines, (HEK293, CHO, HepG2, K562, and Jurkat) were successfully shipped and stored for a minimum of 72 hours and up to 96 hours at ambient temperature, after which, cells were recovered into standard culture conditions. Viability (%) and cell numbers, were examined, before, following the transport/storage period and following the recovery period. In all experiments, cell numbers returned to pretransport/storage concentration within 24-48 hours recovery. Imaging data indicated that HepG2 cells were fully adherent and had established typical growth morphology following 48 hours recovery, which was not seen in cells recovered from cryopreservation. Following recovery, Jurkat cells that had been subjected to a 96 hours transport/storage period, demonstrated a 1.93-fold increase compared with the starting cell number with >95% cell viability. We conclude that CellShip® may represent a viable method for the transportation of mammalian cells for multiple downstream applications in the Life Sciences research sector.

Age-Related Differences in Trunk Kinematics and Interplanar Decoupling with the Pelvis during Gait in Healthy Older versus Younger Men.

This study investigated age-related differences in trunk kinematics during walking in healthy men. Secondary aims were to investigate the covarying effects of physical activity (PA) and lumbar paravertebral muscle (LPM) morphology on trunk kinematics, and the effect of age on interplanar coupling between the trunk and pelvis. Three-dimensional (3D) trunk and pelvis motion data were obtained for 12 older (67.3 ± 6.0 years) and 12 younger (24.7 ± 3.1 years) healthy men during walking at a self-selected speed along a 10 m walkway. Phase-specific differences were observed in the coronal and transverse planes, with midstance and swing phases highlighted as instances when trunk and pelvic kinematics differed significantly (p < 0.05) between the younger group and older group. Controlling for age, fewer significant positive correlations were revealed between trunk and pelvic ranges and planes of motion. LPM morphology and PA were not significant covariates of age-related differences in trunk kinematics. Age-related differences in trunk kinematics were most apparent in the coronal and transverse planes. The results further indicate ageing causes an uncoupling of interplanar upper body movements during gait. These findings provide important information for rehabilitation programmes in older adults designed to improve trunk motion, as well as enable identification of higher-risk movement patterns related to falling.