The effect of growth hormone administration on the regulation of mitochondrial apoptosis in-vivo

The purpose of this study was to determine whether recombinant human growth hormone (rhGH) would show any significant effects on the expression of apoptosis regulating proteins in peripheral blood mononuclear cells (PBMCs). Additionally, the potential for post-transcriptional regulation of gene expression by miRNA was assessed in two cellular compartments, the cytosol and the mitochondria. Ten male subjects were subcutaneously injected with either rhGH (1 mg) or saline (0.9%) for seven consecutive days in a double-blinded fashion. Blood sampling was undertaken prior to treatment administration and over a period of three weeks following treatment cessation. Bcl-2 and Bak gene and protein expression levels were measured in PBMCs, while attention was also directed to the expression of miR-181a and miR-125b, known translational inhibitors of Bcl-2 and Bak respectively. Results showed that rhGH significantly decreased Bak protein concentrations compared to placebo samples for up to 8 days post treatment. While cytosolic miRNA expression was not found to be significantly affected by rhGH, measurement of the expression of miR-125b in mitochondrial fractions showed a significant down-regulation eight days post-rhGH administration. These findings suggest that rhGH induces short-term anti-apoptotic effects which may be partially mediated through a novel pathway that alters the concentration of mitochondrially-associated miRNAs

Recombinant human growth hormone and insulin-like growth factor-1 do not affect mitochondrial derived highly reactive oxygen species production in peripheral blood mononuclear cells under conditions of substrate saturation in-vitro

BACKGROUND: The purpose of this study was to investigate the mitochondrial effects exerted by physiological and supra-physiological concentrations of recombinant human growth hormone (rhGH) and recombinant insulin-like growth factor-1 (rIGF-1) under conditions of substrate saturation in peripheral blood mononuclear cells (PBMCs). METHODS: PBMCs from healthy male subjects were treated with either rhGH, at concentrations of 0.5, 5 and 50 μg/L, or rIGF-1 at concentrations of 100, 300 and 500 μg/L for 4 h. Mitochondrial membrane potential (Δψ(m)) and mitochondrial levels of highly reactive oxygen species (hROS) were subsequently analysed. This analysis was performed by flow cytometry in digitonin permeabilized cells, following treatment with saturating concentrations of various respiratory substrate combinations and the use of specific electron transport chain (ETC.) complex inhibitors, enabling control over both the sites of electron entry into the ETC. at complexes I and II and the entry of electrons from reduced carriers involved in β-oxidation at the level of ubiquinol. RESULTS: Neither rhGH nor rIGF-1 exerted any significant effect on Δψ(m) or the rate of hROS production in either lymphocyte or monocyte sub-populations under any of the respiratory conditions analysed. CONCLUSION: That neither hormone was capable of attenuating levels of oxidative stress mediated via either complex I linked respiration or lipid-derived respiration could have serious health implications for the use of rhGH in healthy individuals, which is frequently associated with significant increases in the bioavailability of free fatty acids (FFA). Such elevated supplies of lipid-derived substrates to the mitochondria could lead to oxidative damage which would negatively impact mitochondrial function

Erythrocytes in multiple sclerosis: forgotten contributors to the pathophysiology?

Multiple sclerosis (MS) is an autoimmune disease characterised by lymphocytic infiltration of the central nervous system and subsequent destruction of myelin and axons. On the background of a genetic predisposition to autoimmunity, environmental triggers are assumed to initiate the disease. The majority of MS research has focused on the pathological involvement of lymphocytes and other immune cells, yet a paucity of attention has been given to erythrocytes, which may play an important role in MS pathology. The following review briefly summarises how erythrocytes may contribute to MS pathology through impaired antioxidant capacity and altered haemorheological features. The effect of disease-modifying therapies on erythrocytes is also reviewed. It may be important to further investigate erythrocytes in MS, as this could broaden the understanding of the pathological mechanisms of the disease, as well as potentially lead to the discovery of novel and innovative targets for future therapies

Decrease of DHEA-S concentration succeeding a micro-dose thumb exertion: Mood-state determinants reflect stress-biomarker responses

Background: The present study examined the effect of a micro-dose of resistance-exercise on serum DHEA-S, IL-6 and mood-state determinants. Potential relationships between mood and the biomarkers were also studied with the aim of directing research on non-invasive exercise-monitoring methods. Methods: 30 male participants (20 weightlifting-trained; 10 untrained) were separated into 3 groups of 10: weightlifting experimental (WLEXP); untrained experimental (UTEXP); weightlifting placebo (WLPLA). WLEXP and UTEXP performed four 60-s isometric thumb exertions separated by 60-s rest intervals in a single-blinded placebo-controlled study. Participants were assessed over a 60-min post-intervention recovery period for changes in serum DHEA-S and IL-6, and mood-state determinants (vigour, tension, fatigue). Results: DHEA-S changed in UTEXP only; a decrease from 20- to 60-min post-exercise (Δ36.9 %, p < 0.01). DHEA-S remained below baseline at the final time-point (Δ35.3 %, p = 0.012). Tension decreased immediately post-exercise in WLEXP (Δ86.7 %, p = 0.022), whereas UTEXP showed a delayed decrease which continued up to 60-min post-intervention (Δ100 %, p < 0.01). Relative to fatigue scores recorded immediately post-exercise, WLEXP decreased within the first 10-min post-intervention (Δ22.2 %, p < 0.01) whereas UTEXP showed a delayed decrease evident at 20-min post-intervention (Δ25 %, p < 0.01). Serum IL-6 and vigour scores remained unchanged across groups (p > 0.05) and WLPLA did not change for any measured variable (p > 0.05). Conclusions: The authors conclude that a micro-dose of resistance-exercise can reduce serological DHEA-S concentration within 60-min of exercise cessation. Additionally, mood-state assessment in untrained individuals can be considered for non-invasively indicating exercise-induced concentration changes in the stress biomarker, DHEA-S, providing prospects for the development of safer, more sophisticated exercise-monitoring practice.Griffith Health, School of Allied Health SciencesFull Tex

Investigation of the NOTCH3 and TNFSF7 Genes on C19p13 as Candidates for Migraine

To investigate the migraine locus around the C19p13 region through analysis of the NOTCH3 gene (C19p13.2-p13.1), previously shown to be a gene involved in CADASIL and the TNFSF7 gene (C19p13), homologous to the ligands of TNF-alpha and TNF-beta, genes that have previously been associated with migraine. The NOTCH3 gene was analysed by sequencing all exons with known CADASIL mutations in a typical (non-familial hemiplegic) migraine family (MF1) that has previously been shown to be linked to C19p13. The TNFSF7 gene was investigated through SNP association analysis using a matched case-control migraine population. NOTCH3 gene sequencing results for affected members of MF1 proved to be negative for all known sequence variants giving rise to mutations for CADASIL. TNFSF7 gene chi-square results showed non-significant P values across all populations tested against controls, except for the MO subgroup which displayed a possible association with the TNFSF7 SNP (genotype, allele analysis P = 0.036, P = 0.017 respectively). Our results suggest that common migraine is not caused by any known CADASIL mutations in the NOTCH3 gene of interest. However, the TNFSF7 gene displayed signs of involvement in a MO affected population and indicates that further independent studies of this marker are warranted