Assessment of Anabolic Androgenic Steroids Use Among Professional CrossFit® Athletes: Motives, Perception, and Safety.

BACKGROUND: Anabolic androgenic steroids (AAS) are traditionally used for the treatment/control of various diseases; however, they are being used for non-therapeutic and indiscriminate purposes to enhance sports performance and physical appearance. This study aimed to assess the prevalence and associated factors of AAS use among professional CrossFit® competitors. METHODS: We conducted an observational cross-sectional survey in which an anonymous questionnaire was applied to professional CrossFit® athletes. RESULTS: The prevalence of AAS usage was 33.3%. Most users were male (74.2%), aged between 30 and 39 years (51.6%), with completed higher education (83.9%), and had been training for more than 5 years (77.4%); the primary motivation for AAS use was performance enhancement (77.4%). Individuals who were older (p < 0.05) and more experienced in competitions (p < 0.01) are more likely to use AAS. Testosterone was the most employed AAS (71.0%); CrossFit® athletes typically used an average of 2 different AAS. The majority of users had notably sought advice from a physician (74.2%), and AAS were acquired from either drugstores (80.6%) or through illicit channels (29.0%). Moreover, 61.3% of AAS users reported experiencing adverse effects. CONCLUSIONS: Our results demonstrated a higher prevalence of AAS users among professional competitors in CrossFit® compared to the general population; older age and greater experience in official competitions were decisive factors for a greater inclination toward AAS use. A significant percentage of athletes seek drugs through illegal channels. Despite the majority of users experiencing adverse effects, athletes report satisfaction with use, believing that the benefits still outweigh the drawbacks.

Effects of physical training on hypothalamic neuronal activation and expressions of vasopressin and oxytocin in SHR after running until fatigue.

To assess the influence of physical training on neuronal activation and hypothalamic expression of vasopressin and oxytocin in spontaneously hypertensive rats (SHR), untrained and trained normotensive rats and SHR were submitted to running until fatigue while internal body and tail temperatures were recorded. Hypothalamic c-Fos expression was evaluated in thermoregulatory centers such as the median preoptic nucleus (MnPO), medial preoptic nucleus (mPOA), paraventricular nucleus of the hypothalamus (PVN), and supraoptic nucleus (SON). The PVN and the SON were also investigated for vasopressin and oxytocin expressions. Although exercise training improved the workload performed by the animals, it was reduced in SHR and followed by increased internal body temperature due to tail vasodilation deficit. Physical training enhanced c-Fos expression in the MnPO, mPOA, and PVN of both strains, and these responses were attenuated in SHR. Vasopressin immunoreactivity in the PVN was also increased by physical training to a lesser extent in SHR. The already-reduced oxytocin expression in the PVN of SHR was increased in response to physical training. Within the SON, neuronal activation and the expressions of vasopressin and oxytocin were reduced by hypertension and unaffected by physical training. The data indicate that physical training counterbalances in part the negative effect of hypertension on hypothalamic neuronal activation elicited by exercise, as well as on the expression of vasopressin and oxytocin. These hypertension features seem to negatively influence the workload performed by SHR due to the hyperthermia derived from the inability of physical training to improve heat dissipation through skin vasodilation.

Evaluation of Rumen Fermentation and Microbial Adaptation to Three Red Seaweeds Using the Rumen Simulation Technique.

Several red seaweeds have been shown to inhibit enteric CH4 production; however, the adaptation of fermentation parameters to their presence is not well understood. The objective of this study was to examine the effect of three red seaweeds (Asparargopsis taxiformis, Mazzaella japonica, and Palmaria mollis) on in vitro fermentation, CH4 production, and adaptation using the rumen simulation technique (RUSITEC). The experiment was conducted as a completely randomized design with four treatments, duplicated in two identical RUSITEC apparatus equipped with eight fermenter vessels each. The four treatments included the control and the three red seaweeds added to the control diet at 2% diet DM. The experimental period was divided into four phases including a baseline phase (d 0-7; no seaweed included), an adaptation phase (d 8-11; seaweed included in treatment vessels), an intermediate phase (d 12-16), and a stable phase (d 17-21). The degradability of organic matter (p = 0.04) and neutral detergent fibre (p = 0.05) was decreased by A. taxiformis during the adaptation phase, but returned to control levels in the stable phase. A. taxiformis supplementation resulted in a decrease (p < 0.001) in the molar proportions of acetate, propionate, and total volatile fatty acid (VFA) production, with an increase in the molar proportions of butyrate, caproate, and valerate; the other seaweeds had no effect (p > 0.05) on the molar proportions or production of individual VFA. A. taxiformis was the only seaweed to suppress CH4 production (p < 0.001), with the suppressive effect increasing (p < 0.001) across phases. Similarly, A. taxiformis increased (p < 0.001) the production of hydrogen (H2, %, mL/d) across the adaptation, intermediate, and stable phases, with the intermediate and stable phases having greater H2 production than the adaptation phase. In conclusion, M. japonica and P. mollis did not impact rumen fermentation or inhibit CH4 production within the RUSITEC. In contrast, we conclude that A. taxiformis is an effective CH4 inhibitor and its introduction to the ruminal environment requires a period of adaptation; however, the large magnitude of CH4 suppression by A. taxiformis inhibits VFA synthesis, which may restrict the production performance in vivo.

The role of peripheral transient receptor potential vanilloid 1 channels in stress-induced hyperthermia in rats subjected to an anxiogenic environment.

Anxiety resulting from psychogenic stimuli elicit stress-induced hyperthermia in rats, often called "psychogenic fever", which is part of a coordinated response to situations seen as novel or distressing. Brain transient receptor potential vanilloid 1 (TRPV1) channels modulate both thermoregulation and animal behavior; however, the role of peripheral TRPV1 channels in regulating these responses during exposure to an anxiogenic environment has not been determined. Thus, the present study aimed to investigate the involvement of abdominal TRPV1 channels in stress-induced hyperthermia and behavior in rats subjected to an unconditioned anxiety test. Desensitized rats (peripheral desensitization of TRPV1 channels with resiniferatoxin; RTX) and their respective controls were subjected to a 15-min open field (OF) test. The core body temperature (Tcore), tail skin temperature (Tskin), and rats' movements inside the arena were recorded. The OF test induced a similar increase in Tcore in both groups throughout the exposure time; however, at the recovery period, the RTX-treated rats had a slower reduction in Tcore due to lower tail skin heat loss. Tskin decreased significantly in both groups during exposure to OF but, during recovery, the RTX-treated rats showed impaired skin vasodilation. Also, RTX-treated rats entered fewer times and spent less time in the OF center square, suggesting an anxiety-related behavior. Our findings indicate that, under stressful conditions, peripheral TRPV1 channels modulate thermoregulatory and behavioral responses. The TRPV1 desensitization induces a more prolonged hyperthermic response due to lower cutaneous heat dissipation, alongside a more evident anxiety-like behavior in rats subjected to the OF apparatus.

Survey of off-label prescribing in a university paediatric outpatient in Brazil.

Ours is a cross-sectional, descriptive, retrospective study evaluating the extent of off-label prescribing for patients attending a university paediatric outpatient department in Goiás, Brazil. 391 patients were treated in the outpatient, and 668 medicines were prescribed. Of these, 70.4% followed the terms of the marketing authorization; 0.3% were unlicenced, and 11% were off-label. Dose was the main factor in off-label prescribing. Infants (0-2 years) received 37.8% of the off-label prescriptions. Vitamins and drugs for the treatment of respiratory diseases were the most prevalent culprits. Of the total prescriptions, 23 different drugs were defined as off-label. Salbutamol was the most prescribed (41.9%). Owing to practical and legal difficulties in carrying out clinical trials, medicines are inadequately studied in children; cooperation between industry, regulatory authorities, and healthcare professionals is required to improve treatment safety. Our results may help guide clinical researcher on off-label prescripting in future trials.

Early maternal separation alters the activation of stress-responsive brain areas in adulthood.

The expression of c-Fos protein has been extensively used as a marker of neuronal activation in response to stressful stimuli. Early maternal separation (MS) is a model of early life adversity that affects the responsiveness of the brain areas to stressors. Thus, this study examined the impact of early MS on activating stress-responsive areas in the brain of adult rats in response to physical (ether) or psychological (restraint) stressors. Male pups were divided for the MS or non-handled (NH) groups. The MS was carried out daily between the 2nd and 14th day of postnatal life and consisted in removing the dams from the cage for 180 min. The rats were then subjected to experimental protocols of restraint or ether exposure at 10-12 weeks old. The rats were anesthetized 90 min after exposure to the stressors, and their brains were prepared for immunohistochemical analysis of c-Fos immunoreactive (c-Fos-ir) neurons in the hypothalamic paraventricular nucleus (PVN), supraoptic nucleus (SON), medial preoptic area (MPA), medial amygdaloid nucleus (MeA), locus coeruleus (LC), and nucleus of the solitary tract (NST). The MS-group presented 86%, 125%, 73%, 56%, and 137% higher c-Fos-ir neurons in the LC, PVN, SON, MPA, and MeA, respectively, compared to NH-group in response to the restraint stressor. In addition, the MS-group presented 180%, 137%, 170%, and 138% higher c-Fos-ir neurons for the ether exposure in the LC, PVN, MPA, and MeA, respectively. Our results show a greater increase in neuronal activation in the MS group, indicating that early life adversity can induce reprogramming in the brain response to stress in adulthood.

Exercise capacity in different stages of hypertension in spontaneously hypertensive rats.

BACKGROUND: The aim of the present study was to investigate the exercise capacity of hypertensive rats at different stages of development of hypertension and to determine the most suitable index to evaluate the exercise capacity in different strains. METHODS: Male spontaneously hypertensive rat (SHR) and normotensive Wistar rats (NWR) of 5, 8, 12 and 16 weeks were submitted to the exercise capacity test. The exercise running time was measured and the workload was calculated. RESULTS: Normotensive and hypertensive rats when assess the exercise capacity by exercise running time exhibited a reduction in exercise performance over time. Moreover, hypertensive rats showed lower exercise capacity compared to normotensive control when analyzed by workload. CONCLUSIONS: The present results indicate that hypertensive rats exhibit reduced exercise capacity compared to normotensive rats regardless of age assessed. Beside that, in experiments with strains with different body mass the most reliable index to assess exercise capacity is workload.

Effects of Maternal Dietary Cottonseed on the Profile of Minerals in the Testes of the Lamb.

The use of cotton co-products in animal feed is restricted by the presence of gossypol, which is a toxic and highly reactive molecule of complex minerals. In mammals, part of the offspring phenotype is influenced by dam nutrition. The aim of this study was to investigate the effect of ewe diet, with and without cottonseed (gossypol), on the testicular development of lambs from birth to weaning through the assessment of lamb live weight (LW), macro and histological morphology of testes, and mass fraction of chemical elements by neutron activation analysis (NAA) and microprobe X-ray fluorescence spectroscopy, as well as the multielement distribution map in the testes. Eighteen lambs were used with an average LW at birth of 4 ± 1.0 kg. All lambs were offspring of Santa Inês ewes, fed on ration either with or without cottonseed during mating, gestation, and lactation, thus forming two treatments: control group (C) without cottonseed and treatment group with cottonseed (G). The animals were weighed from birth to 60 days of age, at fortnightly intervals. At 60 days of age, the lambs were orchidectomized to collect their testes for macroscopy, histological, neutron activation, and X-ray fluorescence analysis. Besides dry matter (DM), protein, ether extract (EE), calcium (Ca), and potassium (K) were higher in the ewe milk from the C group compared with the G group (P < 0.05). Lambs from the C group showed higher LW from 45 days onwards, as well as higher average daily gain when compared with the G group (P < 0.05). They also presented higher testicular weight, volume, length, width, as well as tubule and lumen diameters compared with lambs from the G treatment (P < 0.05). Multielementary NAA revealed higher mass fractions of rubidium (Rb), selenium (Se), and cesium (Cs) in the testes of lambs from the C group when compared with the G group, while G showed higher zinc (Zn) content (P < 0.05). No differences between treatments were found for element levels and distribution using X-ray fluorescence microanalysis (P > 0.05). In conclusion, the maternal cottonseed diet compromised performance and testes development of the lambs and reduce the content of Se in the testes.

The time-course of thermoregulatory responses during treadmill running is associated with running duration-dependent hypothalamic neuronal activation in rats.

This study evaluated the hypothalamic neuronal activation during exercise and investigated whether this activation is related to heat storage and exercise duration. Rats were subjected to a treadmill running that was interrupted at three different moments: (1) at the early phase, when minimal heat dissipation occurred due to tail vasoconstriction and the tail skin temperature (Tskin) reached its nadir; (2) at the steady-state phase, when both the Tskin and core body temperature (Tcore) plateaued at a high level (~ 20 min); and (3) at fatigue, when Tcore and Tskin were still elevated. c-Fos expression in the medial and ventromedial preoptic areas (mPOA and vmPOA), median preoptic nucleus (MnPO), paraventricular and supraoptic nucleus (PVN and SON), and septohypothalamic nucleus (SHy) was determined. Exercise increased the expression of c-Fos in all brain areas, but with different activation patterns of activation. c-Fos expression in the SHy and vmPOA was similar in all exercising groups, while in the mPOA, MnPO, and PVN, c-Fos expression gradually increased during exercise. Increased c-Fos in the SON was only evident after 20 min of exercise. Neuronal activation in the mPOA, MnPO, PVN, and SON was positively correlated with both exercise duration and heat storage. Our findings indicate that with the exception of SON, the brain areas analyzed are recruited following small changes in Tcore (~ 0.5 °C), while the SON is recruited only when Tcore reaches higher values (greater than 1.0 °C increase). c-Fos expression in the PVN, mPOA, MnPO, and SON is also influenced by exercise duration, which does not occur in the SHy and vmPOA.

Fiber fractions, multielemental and isotopic composition of a tropical C4 grass grown under elevated atmospheric carbon dioxide.

BACKGROUND: Brazil has the largest commercial herd of ruminants with approximately 211 million head, representing 15% of world's beef production, in an area of 170 million hectares of grasslands, mostly cultivated with Brachiaria spp. Although nutrient reduction due to increased atmospheric carbon dioxide (CO2) concentration has already been verified in important crops, studies evaluating its effects on fiber fractions and elemental composition of this grass genus are still scarce. Therefore, a better understanding of the effects of elevated CO2 on forage quality can elucidate the interaction between forage and livestock production and possible adaptations for a climate change scenario. The objective of this study was to evaluate the effects of contrasting atmospheric CO2 concentrations on biomass production, morphological characteristics, fiber fractions, and elemental composition of Brachiaria decumbens (cv. Basilisk). METHODS: A total of 12 octagonal rings with 10 m diameter were distributed in a seven-ha coffee plantation and inside each of them, two plots of 0.25 m2 were seeded with B. decumbens (cv. Basilisk) in a free air carbon dioxide enrichment facility. Six rings were kept under natural conditions (≈390 µmol mol-1 CO2; Control) and other six under pure CO2 flux to achieve a higher concentration (≈550 µmol mol-1 CO2; Elevated CO2). After 30 months under contrasting atmospheric CO2 concentration, grass samples were collected, and then splitted into two portions: in the first, whole forage was kept intact and in the second portion, the leaf, true stem, inflorescence and senescence fractions were manually separated to determine their proportions (%). All samples were then analyzed to determine the fiber fractions (NDF, hemicellulose, ADF, cellulose, and Lignin), carbon (C), nitrogen (N), potassium (K), calcium (Ca), sulfur (S), phosphorus (P), iron (Fe), and manganese (Mn) contents and N isotopic composition. RESULTS: Elevated atmospheric CO2 concentration did not influence biomass productivity, average height, leaf, stem, senescence and inflorescence proportions, and fiber fractions (p > 0.05). Calcium content of the leaf and senescence portion of B. decumbens were reduced under elevated atmospheric CO2 (p < 0.05). Despite no effect on total C and N (p > 0.05), lower C:N ratio was observed in the whole forage grown under elevated CO2 (p < 0.05). The isotopic composition was also affected by elevated CO2, with higher values of δ15N in the leaf and stem portions of B. decumbens (p < 0.05). DISCUSSION: Productivity and fiber fractions of B. decumbens were not influenced by CO2 enrichment. However, elevated CO2 resulted in decreased forage Ca content which could affect livestock production under a climate change scenario.

Obesity, Inflammation, and Exercise Training: Relative Contribution of iNOS and eNOS in the Modulation of Vascular Function in the Mouse Aorta.

BACKGROUND: The understanding of obsesity-related vascular dysfunction remains controversial mainly because of the diseases associated with vascular injury. Exercise training is known to prevent vascular dysfunction. Using an obesity model without comorbidities, we aimed at investigating the underlying mechanism of vascular dysfunction and how exercise interferes with this process. METHODS: High-sugar diet was used to induce obesity in mice. Exercise training was performed 5 days/week. Body weight, energy intake, and adipose tissues were assessed; blood metabolic and hormonal parameters were determined; and serum TNFα was measured. Blood pressure and heart rate were assessed by plethysmography. Changes in aortic isometric tension were recorded on myograph. Western blot was used to analyze protein expression. Nitric oxide (NO) was evaluated using fluorescence microscopy. Antisense oligodeoxynucleotides were used for inducible nitric oxide synthase isoform (iNOS) knockdown. RESULTS: Body weight, fat mass, total cholesterol, low-density lipoprotein cholesterol fraction, insulin, and leptin were higher in the sedentary obese group (SD) than in the sedentary control animals (SS). Exercise training prevented these changes. No difference in glucose tolerance, insulin sensitivity, blood pressure, and heart rate was found. Decreased vascular relaxation and reduced endothelial nitric oxide synthase (eNOS) functioning in the SD group were prevented by exercise. Contractile response to phenylephrine was decreased in the aortas of the wild SD mice, compared with that of the SS group; however, no alteration was noted in the SD iNOS(-/-) animals. The decreased contractility was endothelium-dependent, and was reverted by iNOS inhibition or iNOS silencing. The aortas from the SD group showed increased basal NO production, serum TNFα, TNF receptor-1, and phospho-IκB. Exercise training attenuated iNOS-dependent reduction in contractile response in high-sugar diet-fed animals, decreased iNOS expression, and increased eNOS expression. CONCLUSION: Obesity caused endothelium dysfunction, TNFα, and iNOS pathway up-regulation, decreasing vascular contractility in the obese animals. Exercise training was an effective therapy to control iNOS-dependent NO production and to preserve endothelial function in obese individuals.

Central blockade of nitric oxide transmission impairs exercise-induced neuronal activation in the PVN and reduces physical performance.

The blockade of central nitric oxide (NO) signaling modifies the thermoregulatory and metabolic adjustments that occur during exercise, thereby impairing physical performance. However, the brain areas involved in this response remain unknown. Nitrergic neurons are present in the hypothalamic areas that are activated during exercise and participate in autonomic and neuroendocrine responses, such as, the hypothalamic paraventricular nucleus (PVN) and the supraoptic nucleus (SON). To investigate whether brain NO signaling affects thermoregulation during exercise through the activation of hypothalamic neurons, rats underwent acute submaximal treadmill exercise (18 mmin(-1), 5% inclination) until fatigue received an intracerebroventricular injection of 1.43 µmol Nω-nitro-l-arginine metil ester (L-NAME), a nitric oxide synthase inhibitor, or saline (SAL). Skin tail temperature (Tsk) and internal body temperature (Ti) were continuously recorded and c-Fos expression was determined in the PVN and the SON. L-NAME treatment reduced physical performance by 48%, which was positively correlated with tail vasodilation capacity, which was reduced by 28%, and negatively correlated with heat storage rate (HSR), which was increased by 38%. Physical exercise until fatigue increased the number of c-Fos-immunoreactive (ir) neurons in the PVN and the SON. L-NAME-treatment significantly reduced the exercise-induced c-Fos expression in the PVN, whereas it had no effect in the SON. Interestingly, the number of c-Fos-ir neurons in the PVN was closely correlated with physical performance and inversely associated with HSR. Thus, the inhibition of central NO attenuates neuronal activation induced by exercise in the PVN, impairs the autonomic regulation of heat dissipation, and anticipates the fatigue. Brain NO seems to play a role in exercise performance through the regulation of neuronal activation in the PVN, but not in the SON, although the SON neurons are also activated by running exercise. Moreover, this role in performance mediated by neuronal activation in the PVN can be related with the improvement of thermoregulatory adjustments that occur during exercise.

Treadmill exercise induces neutrophil recruitment into muscle tissue in a reactive oxygen species-dependent manner. An intravital microscopy study.

Intense exercise is a physiological stress capable of inducing the interaction of neutrophils with muscle endothelial cells and their transmigration into tissue. Mechanisms driving this physiological inflammatory response are not known. Here, we investigate whether production of reactive oxygen species is relevant for neutrophil interaction with endothelial cells and recruitment into the quadriceps muscle in mice subjected to the treadmill fatiguing exercise protocol. Mice exercised until fatigue by running for 56.3±6.8 min on an electric treadmill. Skeletal muscle was evaluated by intravital microscopy at different time points after exercise, and then removed to assess local oxidative stress and histopathological analysis. We observed an increase in plasma lactate and creatine kinase (CK) concentrations after exercise. The numbers of monocytes, neutrophils, and lymphocytes in blood increased 12 and 24 hours after the exercise. Numbers of rolling and adherent leukocytes increased 3, 6, 12, and 24 hours post-exercise, as assessed by intravital microscopy. Using LysM-eGFP mice and confocal intravital microscopy technology, we show that the number of transmigrating neutrophils increased 12 hours post-exercise. Mutant gp91phox-/- (non-functional NADPH oxidase) mice and mice treated with apocynin showed diminished neutrophil recruitment. SOD treatment promoted further adhesion and transmigration of leukocytes 12 hours after the exercise. These findings confirm our hypothesis that treadmill exercise increases the recruitment of leukocytes to the postcapillary venules, and NADPH oxidase-induced ROS plays an important role in this process.

[Fungi microbiot of Melipona subnitida Ducke (Hymenoptera: Apidae)]. / Microbiota fúngica de Melipona subnitida Ducke (Hymenoptera: Apidae).

This paper reports on the occurrence of filamentous fungi found on the surface of the bees body from the specie Melipona subnitida Ducke that inhabits rocky places on the semi-arid Northeastern Brazil. Bees with cause of natural death were collected of beehives belonging to the Centro de Multiplicação de Animais Silvestres of the Universidade Federal Rural do Semi-Arido. We found the fungi: Aspergillus sp. 6 (37.5%); Aspergillus niger 2 (12.5%); Penicilium sp. 2 (12.5%); Aspergillus terreus 1 (6.3%); Curvularia sp. 1 (6.35%); Monilia sp. 1 (6.3%); Nigrospora sp. 1 (6.3%); Cladosporium sp. 1 (6.3%); Tricoderma sp. 1 (6.3%).

Microbiota fúngica de Melipona subnitida Ducke (Hymenoptera: Apidae) / Fungi microbiot of Melipona subnitida Ducke (Hymenoptera: Apidae)

O presente trabalho descreve a ocorrência de fungos filamentosos sobre a superfície do corpo de abelhas da espécie Melipona subnitida Ducke, que habita regiões pedregosas do semi-árido nordestino. Abelhas com causa de morte natural foram coletadas de colméias pertencentes ao Centro de Multiplicação de Animais Silvestres da Universidade Federal Rural do Semi-Árido. Foram encontrados os fungos: Aspergillus sp. 6 (37,5 por cento); Aspergillus niger 2 (12,5 por cento); Penicilium sp. 2 (12,5 por cento); Aspergillus terreus 1 (6,3 por cento); Curvularia sp. 1 (6,3 por cento); Monilia sp. 1 (6,3 por cento); Nigrospora sp. 1 (6,3 por cento); Cladosporium sp. 1 (6,3 por cento); Tricoderma sp. 1 (6,3 por cento).

This paper reports on the occurrence of filamentous fungi found on the surface of the beeÆs body from the specie Melipona subnitida Ducke that inhabits rocky places on the semi-arid Northeastern Brazil. Bees with cause of natural death were collected of beehives belonging to the Centro de Multiplicação de Animais Silvestres of the Universidade Federal Rural do Semi-Árido. We found the fungi: Aspergillus sp. 6 (37.5 percent); Aspergillus niger 2 (12.5 percent); Penicilium sp. 2 (12.5 percent); Aspergillus terreus 1 (6.3 percent); Curvularia sp. 1 (6.35 percent); Monilia sp. 1 (6.3 percent); Nigrospora sp. 1 (6.3 percent); Cladosporium sp. 1 (6.3 percent); Tricoderma sp. 1 (6.3 percent).