Submaximal exercise intensities do not provoke variations in plasma magnesium concentration in well-trained euhydrated endurance athletes with no magnesium deficiency.

The purpose of this study was to assess the effect of exercise intensity during an incremental exercise test on plasma Mg concentration in well-trained euhydrated athletes. Twenty-seven well-trained endurance athletes carried out a cycloergometer test: after a warm-up of 10 min at 2.0 W·kg(-1), the workload increased by 0.5 W·kg(-1) every 10 min until exhaustion. Oxygen uptake (VO(2)), blood lactate concentration ([La(-)](b)), catecholamines, and plasma Mg were measured at rest, at the end of each stage and at 3, 5 and 7 minutes post-exercise. Urine specific gravity (U(SG)) was analyzed before and after the test, and subjects drank water ad libitum. Fat oxidation rate (FAT(oxr)), carbohydrate oxidation rate (CHO(oxr)), energy expenditure from fat (EE(FAT)), energy expenditure from carbohydrate (EE(CHO)), and total EE (EE(TOTAL)) were estimated using stoichiometric equations. Plasma Mg concentration at each relative exercise intensity (W·kg(-1)) were compared by means of repeated-measures ANOVA. Pearson's correlations were performed to assess the relationship between variables. The significance level was set at p<0.05. No significant differences were found in U(SG) between before and after the test (1.014±0.004 vs 1.014±0.004 g·cm(-3)). Nor were significant differences found in plasma Mg as a function of the different exercise intensities. Further, no significant correlations were detected between Mg and metabolic variables. In conclusion, acute exercise at a range of submaximal intensities in euhydrated well-trained endurance athletes does not affect plasma Mg concentration, suggesting that the plasma volume plays an important role in Mg homeostasis during exercise.

Plasma trace elements levels are not altered by submaximal exercise intensities in well-trained endurance euhydrated athletes.

The purpose of this study was to assess the effect of relative exercise intensity on various plasma trace elements in euhydrated endurance athletes. Twenty-seven well-trained endurance athletes performed a cycloergometer test: after a warm-up of 10 min at 2.0 W kg⁻¹, workload increased by 0.5 W kg⁻¹ every 10 min until exhaustion. Oxygen uptake, blood lactate concentration ([La⁻](b)), and plasma ions (Zn, Se, Mn and Co) were measured at rest, at the end of each stage, and 3, 5 and 7 min post-exercise. Urine specific gravity (U(SG)) was measured before and after the test, and subjects drank water ad libitum. Fat oxidation (FAT(OXR)), carbohydrate oxidation (CHO(OXR)), energy expenditure from fat (EE(FAT)), from carbohydrates (EE(CHO)) and total EE (EE(T)) were estimated using stoichiometric equations. A repeated measure (ANOVA) was used to compare plasma ion levels at each exercise intensity level. The significance level was set at P<0.05. No significant differences were found in U(SG) between, before, and after the test (1.014±0.004 vs. 1.014±0.004 g cm⁻³) or in any plasma ion level as a function of intensity. There were weak significant correlations of Zn (r=0.332, P<0.001) and Se (r=0.242, P<0.01) with [La⁻](b), but no relationships were established between [La⁻](b), VO2, FAT(OXR), CHO(OXR), EE(FAT), EE(CHO), or EE(T) and plasma ion levels. Acute exercise at different submaximal intensities in euhydrated well-trained endurance athletes does not provoke a change in plasma trace element levels, suggesting that plasma volume plays an important role in the homeostasis of these elements during exercise.

Structural properties of the human respiratory syncytial virus P protein: evidence for an elongated homotetrameric molecule that is the smallest orthologue within the family of paramyxovirus polymerase cofactors.

The oligomeric state and the hydrodynamic properties of human respiratory syncytial virus (HRSV) phosphoprotein (P), a known cofactor of the viral RNA-dependent RNA polymerase (L), and a trypsin-resistant fragment (X) that includes its oligomerization domain were analyzed by sedimentation equilibrium and velocity using analytical ultracentrifugation. The results obtained demonstrate that both P and fragment X are homotetrameric with elongated shapes, consistent with electron micrographs of the purified P protein in which thin rod-like molecules of approximately 12.5 +/- 1.0 nm in length were observed. A new chymotrypsin resistant fragment (Y*) included in fragment X has been identified and purified by gel filtration chromatography. Fragment Y* may represent a minimal version of the P oligomerization domain. Thermal denaturation curves based on circular dichroism data of P protein showed a complex behavior. In contrast, melting data generated for fragments X and particularly fragment Y* showed more homogeneous transitions indicative of simpler structures. A three-dimensional model of X and Y* fragments was built based on the atomic structure of the P oligomerization domain of the related Sendai virus, which is in good agreement with the experimental data. This model will be an useful tool to make rational mutations and test the role of specific amino acids in the oligomerization and functional properties of the HRSV P protein.

Structural analysis of the human respiratory syncytial virus phosphoprotein: characterization of an alpha-helical domain involved in oligomerization.

Human respiratory syncytial virus (HRSV) phosphoprotein (P), an essential cofactor of the viral polymerase, is much shorter (241 aa) than and has no sequence similarity to P of other paramyxoviruses. Nevertheless, bioinformatic analysis of HRSV P sequence revealed a modular organization, reminiscent of other paramyxovirus Ps, with a central structured domain (aa 100-200), flanked by two intrinsically disordered regions (1-99 and 201-241). To test the predicted structure experimentally, HRSV P was purified from cell extracts infected with recombinant vaccinia virus or HRSV. The estimated molecular mass of P by gel filtration (approximately 500 kDa) greatly exceeded the theoretical mass of a homotetramer, proposed as the oligomeric form of native P. Nevertheless, the profile of cross-linked products obtained with purified P resembled that reported by others with P purified from bacteria or mammalian cells. Thus, the shape of HRSV P probably influences its elution from the gel filtration column, as reported for other paramyxovirus Ps. Digestion of purified HRSV P with different proteases identified a trypsin-resistant fragment (X) that reacted with a previously characterized monoclonal antibody (021/2P). N-terminal sequencing and mass spectrometry analysis placed the X fragment boundaries (Glu-104 and Arg-163) within the predicted structured domain of P. Cross-linking and circular dichroism analyses indicated that fragment X was oligomeric, with a high alpha-helical content, properties resembling those of the multimerization domain of Sendai and rinderpest virus P. These results denote structural features shared by HRSV and other paramyxovirus Ps and should assist in elucidation of the HRSV P structure.