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

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

The effect of intense interval exercise on selected immunological, haematological and endocrinological parameters in trained male subjects

Exercise has an established ability to alter human physiological systems. However to this time much of the research in this area, and particularly that in relation to the human immune system, has focused on exercise that has required predominantly aerobic metabolism. Little research has been carried out into the physiological effects of intense interval exercise, commonly used as a training regime by athletes in sports such as track & field, ball sports and swimming. This study investigated the effects of intense interval running on selected physiological parameters, with special reference to the immune system and the function of neutrophils. Trained male subjects performed intense interval running protocols both in the laboratory and on a 400 metre running track. The protocols required significant anaerobic energy production as indicated by the elevated post-exercise whole blood lactate concentrations of 7.6 mmol.L-1 and 14.0 to 16.0 mmol.L -1 relating to the laboratory and "track" running protocols, respectively. Intense interval exercise induced a series of biochemical and morphological changes consistent with the multi-faceted activation of neutrophils. Such activation in vivo may have important implications for both tissue damage induced during, and immunocompetence immediately after, intense interval exercise. A single session of intense interval exercise produced significant perturbation of other physiological systems known to be capable of .altering immune function. This was manifested as a significant elevation in the plasma concentrations of several hormones, minor fluctuations in plasma cytokines concentrations and iron status parameters as well as a characteristic pattern of leucocyte mobilisation. After exercise neutrophils displayed phenotypic changes characteristic of activation in vivo. The plasma concentration of elastase (derived from the primary granules of neutrophils) was elevated significantly at 1 hr post-exercise (p<O.OS), while the expression of complement receptor 3 (:::R3;CD11b/CD18), a ts2-integrin stored intracellularly in association with the secondary granules, was increased significantly after exercise (p<0.01). These changes imply that degranulation of neutrophils in vivo was induced by intense interval exercise. Neutrophil expression of LECAM-1, a member of the selectin "family" of cell adhesion molecules, was significantly decreased after exercise (p<0.01). Neutrophils isolated after exercise showed a reduced ability to generate ROS when stimulated in vitro with phorbol myristate acetate (PMA) and opsonized zymosan (OZ), as assessed by both reduction of ferricytochrome c and by luminol-enhanced chemiluminescence, respectively (both p<O.OS). A key component of the neutrophil's NADPH-oxidase system, p47-phox, translocated to the plasma membrane during exercise. The reduced ability of neutrophils isolated after intense interval exercise to generate ROS appears to constitute a post-exercise "refractory" period and be secondary to both assembly and activation of the NADPH-oxidase system during exercise. Increased plasma concentrations of immunosuppressive hormones cortisol and epinephrine during and after intense interval exercise may accentuate the post-exercise refractory period induced after this form of exercise. Our findings indicate that intense interval exercise induced a multi-faceted activation of neutrophils in vivo and imply that both the increased susceptibility of athletes to infection during and after periods of intense competition and training, and tissue damage induced by exercise may be due to activation of neutrophils in vivo during exercise

γδ T cell response to prolonged heavy endurance exercise

The focus of this study was to assess exercise-induced alterations in circulating γδ T cell subpopulations and memory phenotypes after a prolonged heavy-intensity exercise bout. Ten highly-trained endurance cyclists (mean ± SEM: age 24.0 ± 1.3 years; height 1.81 ± 0.02 m; body mass 73.3 ± 1.8 kg; peak oxygen uptake 60.7 ± 1.5 mL.kg-1.min-1) performed 2 h of cycling exercise at 90% of the second ventilatory threshold. Blood samples were collected before exercise, immediately post-exercise, 1 h, 2 h, 4 h, and 6 h post-exercise. Flow cytometry was used to examine γδ T cell subsets, memory phenotypes and receptor expression. A significant decrease in cell concentration was observed in total γδ T cells and the δ2 subset from pre-exercise to 1 h, 2 h, and 4 h post-exercise. Further analysis of the δ2 subset revealed a significant decrease from pre-exercise to 1 h, 2 h, and 4 h post-exercise in naive δ2 cells, and a significant decrease from pre-exercise to 1 h and 2 h post-exercise in central memory δ2 cells. A significant decrease was observed in γδ T cells expressing CD11ahigh, CD62Lhigh and CD94+ from pre-exercise to 1 h, 2 h, and 4 h post-exercise. Furthermore, a significant decrease was observed from pre-exercise to 1 h post-exercise in CD62Llow and CD94- γδ T cells. These results suggest an exercise-stress-induced redistribution of γδ T cells from the circulation with greater propensity for antigen stimulation, tissue and lymph node homing potential for a duration of 4 h after the cessation of exercise

Atmospheric velocity fields in tepid main sequence stars

The line profiles of the stars with v sin i below a few km/s can reveal direct signatures of local velocity fields (e.g. convection) in stellar atmospheres. This effect is well established in cool main sequence stars, and has been detected and studied in three A stars. This paper reports observations of main sequence B, A and F stars with two goals: (1) to identify additional stars having sufficiently low values of v sin i to search for spectral line profile signatures of local velocity fields, and (2) to explore how the signatures of the local velocity fields in the atmosphere depend on stellar parameters such as effective temperature T_eff and peculiarity type. For stars having T_eff below about 10000 K, we always detect local atmospheric velocity fields indirectly through a non-zero microturbulence parameter, but not for hotter stars. Among the A and F stars in our sample having the sharpest lines, direct tracers of atmospheric velocity fields are found in six new stars. The velocity field signatures identified include asymmetric excess line wing absorption, deeper in the blue line wing than in the red; line profiles of strong lines that are poorly fit by computed profiles; and strong lines that are broader than they should be for the v sin i values deduced from weak lines. These effects are found in both normal and Am stars, but seem stronger in Am stars. These data still have not been satisfactorily explained by models of atmospheric convection, including numerical simulations.Comment: Acepted for publication by Astronomy and Astrophysic

Chemical composition of A and F dwarfs members of the Pleiades open cluster

Abundances of 18 chemical elements have been derived for 16 A (normal and chemically peculiar CP) and 5 F dwarfs members of the Pleiades open cluster in order to set constraints on evolutionary models. The abundances, rotational velocities and microturbulent velocities were derived by iteratively adjusting synthetic spectra to observations at high resolution (R~42000 and R~75000) and high signal-to-noise (S/N) ratios. The abundances obtained do not exhibit any clear correlation with the effective temperature nor the projected rotational velocity. Interestingly, A stars exhibit larger star-to-star variations in C, Sc, Ti, V, Cr, Mn, Sr, Y, Zr and Ba than F stars. F stars exhibit solar abundances for almost all the elements. In A stars, the abundances of Si, Ti and Cr are found to be correlated with that of Fe, the [X/Fe] ratios being solar for these three elements. The derived abundances have been compared to the predictions of published evolutionary models at the age of Pleiades (100 Myr). For the F stars, the predicted slight underabundances of light elements and overabundances of Cr, Fe and Ni are indeed confirmed by our findings. For A stars, the predicted overabundances in iron peak elements are confirmed in a few stars only. The large scatter of the abundances in A stars, already found in the Hyades, Coma Berenices and the UMa group and in field stars appears to be a characteristic property of dwarf A stars. The occurence of hydrodynamical processes competing with radiative diffusion in the radiative zones of the A dwarfs might account for the found scatter in abundances.Comment: 7 pages, 3 figures, accepted in A&

Late stages of the evolution of A-type stars on the main sequence: comparison between observed chemical abundances and diffusion models for 8 Am stars of the Praesepe cluster

Aims. We aim to provide observational constraints on diffusion models that predict peculiar chemical abundances in the atmospheres of Am stars. We also intend to check if chemical peculiarities and slow rotation can be explained by the presence of a weak magnetic field. Methods. We have obtained high resolution, high signal-to-noise ratio spectra of eight previously-classified Am stars, two normal A-type stars and one Blue Straggler, considered to be members of the Praesepe cluster. For all of these stars we have determined fundamental parameters and photospheric abundances for a large number of chemical elements, with a higher precision than was ever obtained before for this cluster. For seven of these stars we also obtained spectra in circular polarization and applied the LSD technique to constrain the longitudinal magnetic field. Results. No magnetic field was detected in any of the analysed stars. HD 73666, a Blue Straggler previously considered as an Ap (Si) star, turns out to have the abundances of a normal A-type star. Am classification is not confirmed for HD 72942. For HD 73709 we have also calculated synthetic Delta-a photometry that is in good agreement with the observations. There is a generally good agreement between abundance predictions of diffusion models and values that we have obtained for the remaining Am stars. However, the observed Na and S abundances deviate from the predictions by 0.6 dex and >0.25 dex respectively. Li appears to be overabundant in three stars of our sample.Comment: Accepted for publication on A&

Spectral disentangling of the triple system DG Leo: orbits and chemical composition

DG Leo is a spectroscopic triple system composed by 3 stars of late-A spectral type, one of which was suggested to be a delta Scuti star. Seven nights of observations at high spectral and high time resolution at the Observatoire de Haute-Provence with the ELODIE spectrograph were used to obtain the component spectra by applying a Fourier transform spectral disentangling technique. Comparing these with synthetic spectra, the stellar fundamental parameters (effective temperature, surface gravity, projected rotation velocity and chemical composition) are derived. The inner binary consists of two Am components, at least one of which is not yet rotating synchronously at the orbital period though the orbit is a circular one. The distant third component is confirmed to be a delta Scuti star with normal chemical composition.Comment: MNRAS (Accepted October 7, 2004

Epidemiology of astrovirus infection in children

Human astrovirus (HAstV) is a major cause of acute diarrhea among children, resulting in outbreaks of diarrhea and occasionally hospitalization. Improved surveillance and application of sensitive molecular diagnostics have further defined the impact of HAstV infections in children. These studies have shown that HAstV infections are clinically milder (diarrhea, vomiting, fever) than infections with other enteric agents. Among the 8 serotypes of HAstV identified, serotype 1 is the predominant strain worldwide. In addition to serotype 1, the detection rate of HAstV types 2 to 8 has increased by using newly developed assays. HAstV is less common compared with other major gastroenteritis viruses, including norovirus and rotavirus; however, it is a potentially important viral etiological agent with a significant role in acute gastroenteritis. A better understanding of the molecular epidemiology and characteristics of HAstV strains may be valuable to develop specific prevention strategies