Histological and scanning electron microscope observations on the developing retina of the cuttlefish (Sepia officinalis Linnaeus, 1758).

In this work we present a detailed study of the major events during retinal histogenesis of the cuttlefish Sepia officinalis from early embryos to newly hatched animals and juveniles. For this purpose, we carried out morphometric and histological analyses using light and scanning electron microscopy. From St19, the first embryonic stage analysed, to St23/24 the embryonic retina is composed of a pseudostratified epithelium showing abundant mitotic figures in the more internal surface. At St24 the first photoreceptor nuclei appear in the presumptive inner segment layer, while an incipient layer of apical processes of the future rhabdomeric layer become visible at St25. From this stage onwards, both the rhabdomeric layer and the inner segment layer increase in size until postnatal ages. In contrast, the width of the supporting cell layer progressively decreases from St25/26 until postnatal ages. S. officinalis embryos hatched in a morphologically advanced state, showing a differentiated retina even in the last stages of the embryonic period. However, features of immaturity are still observable in the retinal tissue during the first postnatal weeks of life, such as the existence of mitotic figures in the apical region of the supporting cell layer and migrating nuclei of differentiating photoreceptors crossing the basal membrane to reach their final location in the inner segment layer. Therefore, postnatal retinal neurogenesis is present in juvenile specimens of S. officinalis.

Retinal Development in a Precocial Bird Species, the Quail (Coturnix coturnix, Linnaeus 1758).

The quail (Coturnix coturnix, Linnaeus 1758), a notable model used in developmental biology, is a precocial bird species in which the processes of retinal cell differentiation and retinal histogenesis have been poorly studied. The purpose of the present research is to examine the retinogenesis in this bird species immunohistochemically and compare the results with those from previous studies in precocial and altricial birds. We found that the first PCNA-negative nuclei are detected at Stage (St) 21 in the vitreal region of the neuroblastic layer, coinciding topographically with the first αTubAc-/Tuj1-/Isl1-immunoreactive differentiating ganglion cells. At St28, the first Prox1-immunoreactive nuclei can be distinguished in the vitreal side of the neuroblastic layer (NbL), but also the first visinin-immunoreactive photoreceptors in the scleral surface. The inner plexiform layer (IPL) emerges at St32, and the outer plexiform layer (OPL) becomes visible at St35-the stage in which the first GS-immunoreactive Müller cells are distinguishable. Newly hatched animals show a well-developed stratified retina in which the PCNA-and pHisH3-immunoreactivies are absent. Therefore, retinal cell differentiation in the quail progresses in the stereotyped order conserved among vertebrates, in which ganglion cells initially appear and are followed by amacrine cells, horizontal cells, and photoreceptors. Müller glia are one of the last cell types to be born. Plexiform layers emerge following a vitreal-to-scleral gradient. Finally, our results suggest that there are no significant differences in the timing of different events involved in retinal maturation between the quail and the chicken, but the same events are delayed in an altricial bird species.

Effects of a Multicomponent Programme for Improving Physical and Psychological Health in Victims of Intimate Partner Violence: Study Protocol for a Randomised Control Trial.

Intimate partner violence (IPV) is a public health problem that psychologically and physically affects the women who suffer from it. In this article, we propose an intervention protocol based on therapeutic tourism through adventure physical activities and psychological therapy that could improve the psychological and physical health of women participants. We propose a randomized study where participants will be separated into two groups, control and experimental, and we will perform measurements including self-concept, self-image, depression, and perceived stress, combining these with physiological measurements of stress level by determining stress-related hormones (Cortisol and DHEA), as well as the overall cost-effectiveness of the program. All data collected at the end of the protocol will be statistically analysed. If the final data are positive and it is feasible, this protocol could be proposed as a measure for the treatment of the sequelae of victims of gender violence.

Comparison of Glycosylated Haemoglobin, Blood Pressure, and Anthropometric Measurements Depending on Gender and Bodyweight State in Adolescents.

Background/Objective: The greatest anthropometric and physiological changes occur during adolescence. Assessment of growth patterns is necessary to prevent future health risks. Aims: To describe the values of glycosylated haemoglobin (HbA1c), systolic (SBP) and diastolic (DBP) blood pressure, triceps skinfold, and abdominal circumference according to gender and age (between 12 and 17 years), as well as explore the differences between body weight conditions. Methods: A descriptive cross-sectional study was carried out, including 4130 adolescents between 12 and 17 years old. SBP and DBP, HbA1c, triceps skinfold, and abdominal circumference were evaluated. Results: Significant differences were observed between males and females for HbA1c (p < 0.001), SBP (p < 0.001), triceps curl (p < 0.001), and abdominal circumference (p < 0.001), independently of the age group. Regardless of gender and age groups, significant differences were observed between overweight/obese and normal-weight adolescents in SBP (p < 0.001), DBP (p < 0.001 to 0.009), triceps skinfold (p < 0.001), and abdominal perimeter (p < 0.001). Conclusions: Our findings revealed higher SBP, DBP, triceps skinfold, and abdominal circumference in overweight/obese adolescents compared to normal-weight adolescents in both genders.

Effects of Active Breaks on Physical Literacy: A Cross-Sectional Pilot Study in a Region of Spain.

Several studies have shown that active breaks (AB) lead to improvements in physical fitness, daily steps taken and participants' health. However, there are no studies that have evaluated how they affect physical literacy (PL). Aims: Therefore, this study examined the effects of a 4-week recreational AB program based on games whose main objective was to improve motivation and motor skills' improvement in PL in schoolchildren. Method: A quasi-experimental pilot study was conducted with PL assessments before and after a 4-week recreational AB program. Results: Fifty-seven schoolchildren (10.28 ± 0.43 years) participated in the study, 29 in the control group and 28 in the experimental group. An improvement in PL was found between the experimental and control groups after the intervention (p = 0.017). Moreover, the experimental group also improved (p < 0.001) PL after the intervention. More specifically, within the domain of PL, improvements were found after the intervention in the experimental group in the domains of physical competence (p < 0.001), motivation and confidence (p < 0.001) and knowledge (p < 0.001) but not in the domain of daily activity (p = 0.051). Conclusion: The application of an AB program based on playful games, during four weeks, produced an increase in scores in the general PL level, as well as in the domains of physical competence, motivation, and knowledge and understanding in schoolchildren.

Descriptive Study about Bodyweight Status of Extremadura Adolescents. Are We Applying the Best Indicator as the Reference Parameter?

Background: There is no agreed criteria that establishes childhood obesity thresholds based on BMI, which may be used to assess adolescent overweight/obesity. This tool has been determined at the most practical and least costly in classifying bodyweight status in adolescents. However, it is an indicator of bodyweight and not adiposity. Aims: To assess bodyweight status of Extremadura adolescents by sex and age using international, national, and regional reference criteria and comparing the different diagnoses criteria. Methods: A descriptive cross-sectional study was conducted with 4130 adolescents (12-17 years). Bodyweight and height were assessed. Results: Pairwise comparisons indicates that the World Health Organization (WHO) classified 542 individuals in a different category compared to Faustino Obergozo (FO), and 1028 individuals with respect to the Extremadura adolescents' percentiles (EX). Moreover, FO classified 684 adolescents in a different category than EX. Despite the concordance in diagnostic criteria (by Cohen's kappa test) reported between the WHO, FO, and EX for all bodyweight categories in both sexes and all age ranges, significant differences were found (assessed by Cochran Q test and McNemar test as post-hoc) between the WHO and FO for all bodyweight proportion except in the thinness category in girls (15-17 years) and boys (12-14 years). Meaningful differences were also obtained comparing WHO and EX for each bodyweight category in all ages and sexes. Comparisons between FO and EX revealed significant differences for all bodyweight categories in all participants except for overweight in girls (12-14 years) and boys (15-17 years) and normal weight and obesity in girls (15-17 years). Conclusions: the WHO, FO, and EX criteria present different outcomes estimating overweight and/or obesity prevalence in adolescents aged between 12 and 17 years. The change from Extremadura criteria to the WHO reference will result in more adolescents being diagnosed as overweight or obese.

The Role of Exercise to Reduce the Impact of Diabetes in the Seminal Quality: A Systematic Review.

BackgroundandObjectives: One of the most relevant consequences of diabetes mellitus is the temporal or complete infertility which can happen in young individuals. Therefore, the current systematic review aimed to investigate the effects of exercise to reduce the impact of Type 2 Diabetes Mellitus (T2DM) in seminal quality and related parameters. MaterialsandMethods: A systematic search was conducted in Pubmed and Web of Science databases following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Guidelines (PRISMA). The inclusion criteria were: (1) the study included at least one experimental and one comparison group, (2) the sample of the study was comprised of humans or animals with diabetes mellitus, (3) an intervention based on physical exercise was conducted, and (4) the study reported variables related to the seminal quality. Results: A total of 115 articles were identified. However, only six accomplished the inclusion and exclusion criteria. This systematic review includes a sample size of 260 participants (180 rats and 80 humans). Intervention ranged from 6 to 14 weeks, with 3-6 days per week. All interventions performed endurance training (50-70% VO2max or maximum heart rate). Physical exercise increased sperm count, motility, and morphology, as well as improved testosterone, Luteinizing Hormone (LH), and Follicle Stimulating Hormone (FSH) levels. Moreover, physical exercise intervention reduced the percentages of sperms with negative Tubular Differentiation Index (TDI) and Spermiogenesis Index (SPI), DNA fragmentation, and also ameliorated the diabetes-induced apoptosis and improved sperm apoptosis index. Conclusions: Physical exercise could ameliorate diabetic pathological effects on sperm quality and related parameters that cause infertility or subfertility conditions. However, further homogeneous studies are needed to confirm these findings.

Stage-specific metabolomic changes in equine oviductal fluid: New insights into the equine fertilization environment.

A repeatable protocol for equine in vitro fertilization (IVF) has remained elusive. This is likely, in part, due to suboptimal composition of capacitation or IVF media that are currently in use. Hence, we aimed to analyse the metabolome of equine oviductal fluid (OF) at the pre- (PRE) and immediate post-ovulatory (PST) stages using proton magnetic resonance spectroscopy (1H NMR). Oviductal fluid from eight PRE and six PST mares were used to prepare a total of five samples per group. A total of 18 metabolites were identified. The five metabolites with the highest concentrations in the OF samples were lactate, myoinositol, creatine, alanine and carnitine. Only fumarate and glycine showed significant differences in their concentrations between PRE and PST OF samples, with higher concentrations in the PST samples. In a preliminary study, stallion spermatozoa (n = 3 ejaculates) were incubated with different concentrations of PST OF from one mare (0, 0.0625, 0.125, 0.25, 0.5 or 1%; v:v). After 4 h of sperm incubation, protein tyrosine phosphorylation (PY) by western blotting, sperm motility, and acrosomal status were evaluated. An increase of PY was observed in sperm from two stallions when treated with 0.0625% and 0.125% of OF; however no change in PY was noted in the other stallion. There were no effects of OF on spermatozoa motility or acrosome status. These results provide the first information on the metabolomics of equine OF at different stages of the estrus cycle, and present the possibility that OF may affect PY in stallion spermatozoa.

AMPK Function in Mammalian Spermatozoa.

AMP-activated protein kinase AMPK regulates cellular energy by controlling metabolism through the inhibition of anabolic pathways and the simultaneous stimulation of catabolic pathways. Given its central regulator role in cell metabolism, AMPK activity and its regulation have been the focus of relevant investigations, although only a few studies have focused on the AMPK function in the control of spermatozoa's ability to fertilize. This review summarizes the known cellular roles of AMPK that have been identified in mammalian spermatozoa. The involvement of AMPK activity is described in terms of the main physiological functions of mature spermatozoa, particularly in the regulation of suitable sperm motility adapted to the fluctuating extracellular medium, maintenance of the integrity of sperm membranes, and the mitochondrial membrane potential. In addition, the intracellular signaling pathways leading to AMPK activation in mammalian spermatozoa are reviewed. We also discuss the role of AMPK in assisted reproduction techniques, particularly during semen cryopreservation and preservation (at 17 °C). Finally, we reinforce the idea of AMPK as a key signaling kinase in spermatozoa that acts as an essential linker/bridge between metabolism energy and sperm's ability to fertilize.

Calmodulin inhibitors increase the affinity of Merocyanine 540 for boar sperm membrane under non-capacitating conditions.

We aimed to test whether the calmodulin (CaM) inhibitors, calmidazolium (CZ) and N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7), can be used to assess lipid disorder by flow cytometry using Merocyanine 540 (M540). Boar spermatozoa were incubated in non-capacitating conditions for 10 min at room temperature with 1 µM CZ, 200 µM W-7, or 1 mM 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br-cAMP). Then, sperm were 1) directly evaluated, 2) centrifuged and washed prior to evaluation, or 3) diluted with PBS prior to evaluation. Direct evaluation showed an increase in high M540 fluorescence in spermatozoa treated with the inhibitors (4.7 ± 1.8 [control] vs. 70.4 ± 4.0 [CZ] and 71.4 ± 4.2 [W-7], mean % ± SD, P < 0.001); washing decreased the percentage of sperm showing high M540 fluorescence for W-7 (4.8 ± 2.2, mean % ± SD) but not for CZ (69.4 ± 3.9, mean % ± SD, P < 0.001), and dilution showed an increase in high M540 fluorescence for both CZ and W-7; 8-Br-cAMP did not induce a rise in sperm showing high M540 fluorescence. Therefore, special care must be taken when M540 is used in spermatozoa with CaM inhibitors.

AMP-activated kinase in human spermatozoa: identification, intracellular localization, and key function in the regulation of sperm motility.

AMP-activated kinase (AMPK), a protein that regulates energy balance and metabolism, has recently been identified in boar spermatozoa where regulates key functional sperm processes essential for fertilization. This work's aims are AMPK identification, intracellular localization, and their role in human spermatozoa function. Semen was obtained from healthy human donors. Sperm AMPK and phospho-Thr172-AMPK were analyzed by Western blotting and indirect immunofluorescence. High- and low-quality sperm populations were separated by a 40%-80% density gradient. Human spermatozoa motility was evaluated by an Integrated Semen Analysis System (ISAS) in the presence or absence of the AMPK inhibitor compound C (CC). AMPK is localized along the human spermatozoa, at the entire acrosome, midpiece and tail with variable intensity, whereas its active form, phospho-Thr172-AMPK, shows a prominent staining at the acrosome and sperm tail with a weaker staining in the midpiece and the postacrosomal region. Interestingly, spermatozoa bearing an excess residual cytoplasm show strong AMPK staining in this subcellular compartment. Both AMPK and phospho-Thr172-AMPK human spermatozoa contents exhibit important individual variations. Moreover, active AMPK is predominant in the high motility sperm population, where shows a stronger intensity compared with the low motility sperm population. Inhibition of AMPK activity in human spermatozoa by CC treatment leads to a significant reduction in any sperm motility parameter analyzed: percent of motile sperm, sperm velocities, progressivity, and other motility coefficients. This work identifies and points out AMPK as a new molecular mechanism involved in human spermatozoa motility. Further AMPK implications in the clinical efficiency of assisted reproduction and in other reproductive areas need to be studied.