Sport,doping and male fertility

It is universally accepted that lifestyle interventions are the first step towards a good overall, reproductive and sexual health. Cessation of unhealthy habits, such as tobacco, alcohol and drug use, poor nutrition and sedentary behavior, is suggested in order to preserve/improve fertility in humans. However, the possible risks of physical exercise per se or sports on male fertility are less known. Being “fit” does not only improve the sense of well-being, but also has beneficial effects on general health: in fact physical exercise is by all means a low-cost, high-efficacy method for preventing or treating several conditions, ranging from purely physical (diabetes and obesity) to psychological (depression and anxiety), highly influencing male reproduction. If male sexual and reproductive health could be positively affected by a proper physical activity, inadequate bouts of strength – both excessive intensity and duration of exercise training – are more likely to have detrimental effects. In addition, the illicit use of prohibited drugs (i.e. doping) has reached pandemic proportions, and their actions, unfortunately very often underestimated by both amateur and professional athletes, are known to disrupt at different levels and throughout various mechanisms the male hypothalamic-pituitary-gonadal axis, resulting in hypogonadism and infertility.     

An algorithm to analyse the hydrolysis pathway of peptides and proteins by sequence analyses of unfractionated digestion mixtures

We have designed and implemented on a personal computer a programfor identifying and quantifying the fragments present in a peptidemixture obtained by hydrolysing a polypeptide of known sequenceusing digesting agents. The qualitative data utilized by themain algorithm consist of the target sequence of the intactmolecule and the amino acid residues identified at each stepof the automatic sequence analysis of the unfractionated digestionmixture. In this way, the sequence of each fragment presentin the mixture is quickly reconstructed. Furthermore, if thequantitative data of the amino acid residues identified at eachstep of the sequence analysis are utilized, the program willcorrelate the sequence of each fragment to its amount. We furnishan example of the application intended for the rapid identificationand characterization of the extracellular proteinases producedby a basidiomycete fungus, utilizing the bovine insulin ß3-chainas target substrate. A variety of uses for the method are discussed.     

The characterization of macroH2A beyond vertebrates supports an ancestral origin and conserved role for histone variants in chromatin

Histone variants play a critical role in chromatin structure and epigenetic regulation. These “deviant” proteins have been historically considered as the evolutionary descendants of ancestral canonical histones, helping specialize the nucleosome structure during eukaryotic evolution. Such view is now challenged by 2 major observations: first, canonical histones present extremely unique features not shared with any other genes; second, histone variants are widespread across many eukaryotic groups. The present work further supports the ancestral nature of histone variants by providing the first in vivo characterization of a functional macroH2A histone (a variant long defined as a specific refinement of vertebrate chromatin) in a non-vertebrate organism (the mussel Mytilus) revealing its recruitment into heterochromatic fractions of actively proliferating tissues. Combined with in silico analyses of genomic data, these results provide evidence for the widespread presence of macroH2A in metazoan animals, as well as in the holozoan Capsaspora, supporting an evolutionary origin for this histone variant lineage before the radiation of Filozoans (including Filasterea, Choanoflagellata and Metazoa). Overall, the results presented in this work help configure a new evolutionary scenario in which histone variants, rather than modern “deviants” of canonical histones, would constitute ancient components of eukaryotic chromatin.     

Amino acid sequence around a reactive cysteine of yeast alcohol dehydrogenase

The reaction of yeast alcohol dehydrogenase with iodoacetate produces a loss of 90% of the enzymatic activity when 2.2–2.4 equivalents of carboxymethyl groups are incorporated. After CNBr cleavage of the ¹⁴C-carboxymethylated enzyme the more radioactive fragment was maleylated and digested with trypsin. A tryptic peptide, containing about 70% of initial radioactivity, was purified and sequenced. The cysteine found to be more reactive with iodoacetate in our experimental conditions is different from that found in previous works. It also appears to differ from the cysteine residues found to be reactive towards other thiol reagents.     

Influence of sire and sire breed (Gyr versus Holstein) on establishment of pregnancy and embryonic loss in lactating Holstein cows during summer heat stress

Heat stress has negative effects on pregnancy rates of lactating dairy cattle. There are genetic differences in tolerance to heat stress; Bos taurus indicus (B. t. indicus) cattle and embryos are more thermotolerant than Bos taurus taurus (B. t. taurus). In the present study, the effects of sire and sire breed on conception and embryonic/fetal loss rates of lactating Holstein cows during the Brazilian summer were determined. In Experiment 1, cows (n=302) were AI after estrus detection or at a fixed-time with semen from one Gyr (B. t. indicus) or one Holstein sire (B. t. taurus). Pregnancy was diagnosed 80 days after AI. In Experiment 2, cows (n=811) were AI with semen from three Gyr and two Holstein sires. Pregnancy was diagnosed at 30-40 and at 60-80 days after AI. Cows diagnosed pregnant at the first examination but non-pregnant at the second were considered as having lost their embryo or fetus. Data were analyzed by logistic regression. The model considered the effect of sire within breed, sire breed, days postpartum, period of lactation, and AI type (AI after estrus versus fixed-time). There was no effect of the AI type, days postpartum or milk production on conception or embryonic loss rates. The use of Gyr bulls increased pregnancy rate when compared to Holstein bulls [9.1% (60/657) versus 5.0% (23/456), respectively, P=0.008; data from Experiments 1 and 2 combined]. Additionally, in Experiment 2, cows inseminated using semen from sire #4 (Gyr) had lower embryonic loss (10%) when compared with other B. t. indicus (35.3% and 40%) or B. t. taurus sires (18.2% and 38.5%, P=0.03). In conclusion, the use of B. t. indicus sires may result in higher conception rates in lactating Holstein cows during summer heat stress. Moreover, sire can affect embryonic loss and selection of bulls according to this criterion may result in higher parturition rates in lactating Holstein cows.     

Desarrollo de un bioadsorbente laminar con Phanerochaete chrysosporium hipertolerante al cadmio,al níquel y al plomo para el tratamiento de aguas

BackgroundThe use of basidiomycetes for metal removal is an alternative to traditional methods. In this, the biomass acts as a natural ionic exchanger removing metals from solution.ObjectiveTo develop a laminar biosorbent using a basidiomycete fungus resistant to high Cd, Ni and Pb concentrations.MethodsThe tolerance of Trametes versicolor, Pleurotus ostreatus and Phanerochaete chrysosporium was evaluated using increasing concentrations of the heavy metal salts, cadmium sulphate, lead acetate and nickel chloride. A biosorbent system was developed based on polyethylene sheets with a fungal biomass. It was evaluated in bubble columns using synthetic wastewater with the 3 metal salts at a rate of 300 mg/l. Finally, in a complementary experiment using shake flasks, the effect of a higher amount of biomass related to the metal removal efficiency was evaluated.ResultsP. chrysosporium strain was more tolerant to C₄H₆O₄Pb (10,000 mg/l), Cl₂Ni (300 mg/l) and CdSO₄·8H₂O (1,500 mg/l). In a reactor, under non-ligninolytic conditions, the fungus removed 69% of the chemical oxygen demand and produced enzymes such as LiP (0.01 U/l) and MnP (0.6 U/l.). An accumulation of metals in the wall was observed. By increasing the biomass to 1.6 (w/v), the metal biosorption was favored in the mixture (57% Pb, 74% Cd, and 98% Ni) and separately (95% Pb, 60% Cd, and 56% Ni). Competition between Ni and Pb by ligands of the wall was observed.ConclusionA novel laminar system based on P. chrysosporium viable biomass was developed. It has a large surface area and tolerance to high concentrations of Cd, Ni and Pb. It seems to be an alternative for the removal of metals from water.     

Schooltime subjective sleepiness and performance in Italian primary school children

Despite its clinical importance, the issue of the diurnal time course of sleepiness and performance in children remains largely unexplored. The objective of this study is to draw a profile of daytime subjective sleepiness and performance, at simple and complex tasks, in a cohort of Italian primary school children.To this aim, a sample of 316 children (age range: 5–11 years; mean 8.2 ± 1.5) was recruited and sub-divided into three groups, according to age: Group 1 (5–7 years; N = 127), Group 2 (8–9 years; N = 108), Group 3 (10–11 years; N = 81). Subjective sleepiness and simple performance were evaluated, respectively, through the Pictorial Sleepiness Scale and the Simple Reaction Time Task. Executive functions were addressed by means of the “Go/No-Go Task.” Measurements were made in the children’s class three times a day, one day a week over a 3-week period in order to reliably reflect the habitual time course of sleepiness and performance, within the following time intervals: a) 8:30 am–10:30 am; b) 11 am–1 pm; c) 2 pm–4 pm.For the global sample, a significant increase of subjective sleepiness was found at the end of school day (2–4 pm), although at relatively low levels. No significant differences were observed in reaction times across the day, whereas a significant worsening was detected in performance at complex task already since mid-morning. Significant correlations were found between subjective sleepiness and complex performance at all points.Slight age-related differences were found in the time courses of subjective sleepiness: in fact, a significant overday reduction of vigilance levels, from mid-morning onwards, was observed in children aged 5–9 years, but not in the older children (10–11 years). However, the daily time course of both simple and complex performances did not differ among children of the three age groups. Our results show changes in vigilance and cognitive functions across a typical school day in childhood, as well as age-related differences in sleepiness profile, that we suggest to thoroughly consider when conceiving chronopsychological interventions in the school context.     

Less Frequent and Less Severe Flu-Like Syndrome in Interferon Beta-1a Treated Multiple Sclerosis Patients with at Least One Allele Bearing the G>C Polymorphism at Position -174 of the IL-6 Promoter Gene

One of the most common adverse event of interferon beta (IFNβ) therapy for multiple sclerosis is flu-like syndrome (FLS), which has been reportedly related to increased levels of cytokines such as interleukin 6 (IL-6) and tumor necrosis factor-alpha (TNF-α). Average cytokine levels can be affected by single nucleotide polymorphism in the gene promoter regions. To investigate whether IL-6 -174 G>C and TNF-α -376 G>A polymorphisms could be correlated to the incidence of FLS, and whether an anti-inflammatory/antipyretic therapy may influence FLS development, a prospective observational study was performed in 190 treatment naïve, multiple sclerosis patients who started IM IFNβ-1a 30mcg once weekly. The identification of IL-6 -174 G>C and TNF-α -376 G>A polymorphisms was achieved by performing an amplification-refractory mutation system. Serum IL-6 levels were measured using enzyme-linked immunosorbent assay in blood samples taken before therapy and then after the first and last IFNβ-1a injection of the follow-up. FLS-related symptoms were recorded by patients once per week during the first 12 weeks of therapy into a self-reported diary. We found that patients carrying at least one copy of the C allele at position -174 in the promoter of IL-6 gene produced lower levels of IL-6 and were less prone to develop FLS, which was also less severe. On the contrary, the polymorphism of TNF-α had no effect on FLS. Patients taking the first dose of anti-inflammatory/antipyretic therapy in the peri-injection period (within 1 hour) experienced a reduced FLS severity. In conclusion, the study of IL-6 -174 G>C polymorphism would allow the identification of patients lacking the C nucleotide on both alleles who are at risk of a more severe FLS, and may be addressed to a timely and stronger anti-inflammatory/antipyretic therapy for a more effective FLS prevention.     

The Forgotten Role of Central Volume in Low Frequency Oscillations of Heart Rate Variability

The hypothesis that central volume plays a key role in the source of low frequency (LF) oscillations of heart rate variability (HRV) was tested in a population of end stage renal disease patients undergoing conventional hemodialysis (HD) treatment, and thus subject to large fluid shifts and sympathetic activation. Fluid overload (FO) in 58 chronic HD patients was assessed by whole body bioimpedance measurements before the midweek HD session. Heart Rate Variability (HRV) was measured using 24-hour Holter electrocardiogram recordings starting before the same HD treatment. Time domain and frequency domain analyses were performed on HRV signals. Patients were retrospectively classified in three groups according to tertiles of FO normalized to the extracellular water (FO/ECW%). These groups were also compared after stratification by diabetes mellitus. Patients with the low to medium hydration status before the treatment (i.e. 1st and 2nd FO/ECW% tertiles) showed a significant increase in LF power during last 30 min of HD compared to dialysis begin, while no significant change in LF power was seen in the third group (i.e. those with high pre-treatment hydration values). In conclusion, several mechanisms can generate LF oscillations in the cardiovascular system, including baroreflex feedback loops and central oscillators. However, the current results emphasize the role played by the central volume in determining the power of LF oscillations.