Molecular Mechanisms of Hypoxic Responses via Unique Roles of Ras1, Cdc24 and Ptp3 in a Human Fungal Pathogen Cryptococcus neoformans

Cryptococcus neoformans encounters a low oxygen environment when it enters the human host. Here, we show that the conserved Ras1 (a small GTPase) and Cdc24 (the guanine nucleotide exchange factor for Cdc42) play an essential role in cryptococcal growth in hypoxia. Suppressor studies indicate that PTP3 functions epistatically downstream of both RAS1 and CDC24 in regulating hypoxic growth. Ptp3 shares sequence similarity to the family of phosphotyrosine-specific protein phosphatases and the ptp3Δ strain failed to grow in 1% O₂. We demonstrate that RAS1, CDC24 and PTP3 function in parallel to regulate thermal tolerance but RAS1 and CDC24 function linearly in regulating hypoxic growth while CDC24 and PTP3 reside in compensatory pathways. The ras1Δ and cdc24Δ strains ceased to grow at 1% O₂ and became enlarged but viable single cells. Actin polarization in these cells, however, was normal for up to eight hours after transferring to hypoxic conditions. Double deletions of the genes encoding Rho GTPase Cdc42 and Cdc420, but not of the genes encoding Rac1 and Rac2, caused a slight growth retardation in hypoxia. Furthermore, growth in hypoxia was not affected by the deletion of several central genes functioning in the pathways of cAMP, Hog1, or the two-component like phosphorylation system that are critical in the cryptococcal response to osmotic and genotoxic stresses. Interestingly, although deletion of HOG1 rescued the hypoxic growth defect of ras1Δ, cdc24Δ, and ptp3Δ, Hog1 was not hyperphosphorylated in these three mutants in hypoxic conditions. RNA sequencing analysis indicated that RAS1, CDC24 and PTP3 acted upon the expression of genes involved in ergosterol biosynthesis, chromosome organization, RNA processing and protein translation. Moreover, growth of the wild-type strain under low oxygen conditions was affected by sub-inhibitory concentrations of the compounds that inhibit these biological processes, demonstrating the importance of these biological processes in the cryptococcal hypoxia response.     

Emergence of Super Cooperation of Prisoner’s Dilemma Games on Scale-Free Networks

Recently, the authors proposed a quantum prisoner’s dilemma game based on the spatial game of Nowak and May, and showed that the game can be played classically. By using this idea, we proposed three generalized prisoner’s dilemma (GPD, for short) games based on the weak Prisoner’s dilemma game, the full prisoner’s dilemma game and the normalized Prisoner’s dilemma game, written by GPD_W, GPD_F and GPD_N respectively. Our games consist of two players, each of which has three strategies: cooperator (C), defector (D) and super cooperator (denoted by Q), and have a parameter γ to measure the entangled relationship between the two players. We found that our generalised prisoner’s dilemma games have new Nash equilibrium principles, that entanglement is the principle of emergence and convergence (i.e., guaranteed emergence) of super cooperation in evolutions of our generalised prisoner’s dilemma games on scale-free networks, that entanglement provides a threshold for a phase transition of super cooperation in evolutions of our generalised prisoner’s dilemma games on scale-free networks, that the role of heterogeneity of the scale-free networks in cooperations and super cooperations is very limited, and that well-defined structures of scale-free networks allow coexistence of cooperators and super cooperators in the evolutions of the weak version of our generalised prisoner’s dilemma games.     

Fitness improvements of young soccer players after high volume or small sided games interventions

The main goal was to determine anaerobic and aerobic improvement of young soccer players after six-week high volume (HVT) or small sided games (SSG) training intervention. One hundred and one highly trained youth soccer players (16.2 ± 1.3 years) were divided into SSG (n = 51) and HVT groups (n = 50) and according to age into an under sixteen subgroup (U16), under seventeen subgroup (U17), and under nineteen subgroup (U19). The performance was assessed by Yo-Yo intermittent test, Repeated sprint ability test (RSA), and K-test before and after both training interventions. For U16 the SSG group recorded significant improvements in the K-test (0.64 ± 0.56 s; p = .04) and RSA (0.15 ± 0.43 s; p = .01). For U19 the SSG group recorded the same improvements, in the K-test (0.43 ± 0.57 s; p = .007), RSA (0.21 ± 0.22 s; p = .048), and Yo-Yo test (127.25 ± 17.87; p = .049). HVT improved aerobic performance when the Yo-Yo test was significantly better after intervention at U17 (199.00 ± 111.83 m; p = .030), U19 (88.40 ± 66.38 m; p = .049). In total, the HVT group spent 621 min (56.45 ± 5.01 min) of aerobic training and the Small sided game group spent 291 min (26.45 ± 8.61 min) of small sided games focused on aerobic performance. This study showed that both SSG and HVT training interventions were effective for aerobic improvement for the U19 category, but not for younger players. SSG was identified to be more appropriate to fitness development of soccer players.     

Cognitive Functions in Elite and Sub-Elite Youth Soccer Players Aged 13 to 17 Years

Soccer players are required to anticipate and react continuously in a changing, relatively unpredictable situation in the field. Cognitive functions might be important to be successful in soccer. The current study investigated the relationship between cognitive functions and performance level in elite and sub-elite youth soccer players aged 13–17 years. A total of 47 elite youth soccer players (mean age 15.5 years, SD = 0.9) and 41 sub-elite youth soccer players (mean age 15.2 years, SD = 1.2) performed tasks for “higher-level” cognitive functions measuring working memory (i.e., Visual Memory Span), inhibitory control (i.e., Stop-Signal Task), cognitive flexibility (i.e., Trail Making Test), and metacognition (i.e., Delis-Kaplan Executive Function System Design Fluency Test). “Lower-level” cognitive processes, i.e., reaction time and visuo-perceptual abilities, were also measured with the previous tasks. ANOVA’s showed that elite players outscored sub-elite players at the “higher-level” cognitive tasks only, especially on metacognition (p < .05). Using stepwise discriminant analysis, 62.5% of subjects was correctly assigned to one of the groups based on their metacognition, inhibitory control and cognitive flexibility performance. Controlling for training hours and academic level, MANCOVA’s showed differences in favor of the elite youth soccer players on inhibitory control (p = .001), and cognitive flexibility (p = .042), but not on metacognition (p = .27). No differences were found concerning working memory nor the “lower-level” cognitive processes (p > .05). In conclusion, elite youth soccer players have better inhibitory control, cognitive flexibility, and especially metacognition than their sub-elite counterparts. However, when training hours are taken into account, differences between elite and sub-elite youth soccer players remain apparent on inhibitory control and cognitive flexibility in contrast to metacognition. This highlights the need for longitudinal studies to further investigate the importance of “higher-level” cognitive functions for talent identification, talent development and performance in soccer.     

Short-term effects of on-field combined core strength and small-sided games training on physical performance in young soccer players

This study aimed to compare the effects of 6-weeks combined core strength and small-sided games training (SSG_core) vs. small-sided games (SSG) training on the physical performance of young soccer players. Thirty-eight amateur soccer players (age: 16.50 ± 0.51 years) were randomly assigned to either a SSG_core (n = 20) or a SSG group (n = 18). The SSG_core group performed upper and lower body core strength exercises combined with SSG including 2-, 3- and 4-a-sided soccer games third a week. The SSG group performed only the SSG periodization. Baseline and after the 6-week training period the Yo-Yo Intermittent Recovery Test level 1 (YYIRTL-1), 5–20-m sprint test, countermovement jump (CMJ), squat jump (SJ), triple-hop distance (THD), zigzag agility with ball (ZAWB) and without ball (ZAWOB), three corner run test (TCRT) and Y-balance test. The SSG_core group demonstrated meaningful improvements in 20 m sprint time (SSG_core: -9.1%, d = 1.42; SSG: -4.4%, d = 0.76), CMJ (SSG_core: 11.4%, d = 2.67; SSG: -7.7%, d = 1.43), SJ (SSG_core: 12.0%, d = 2.14; SSG: 5.7%, d = 1.28), THD (SSG_core: 5.0%, d = 1.39; SSG: 2.7%, d = 0.52) and TCRT (SSG_core: -3.7%, d = 0.69; SSG: -1.9%, d = 0.38). Furthermore, the SSG_core group demonstrated meaningfully higher improvement responses in both leg balance score (d = ranging from 2.11 to 2.75) compared with SSG group. These results suggest that the inclusion of core strength training to a SSG periodization is greatly effective to improve speed and strength-based conditioning in young soccer players.     

Comparison of the physiological responses to different small-sided games in elite young soccer players

The purpose of this study was to compare the blood lactate (La-), heart rate (HR) and percentage of maximum HR (%HRmax) responses among the small-sided games (SSGs) in elite young soccer players. Sixteen players (average age 15.7 6 0.4 years; height 176.8 6 4.6 cm; body mass 65.5 6 5.6 kg; VO2max 53.1 6 5.9 ml · kg(-1) · min(-1); HRmax 195.9 6 7.4 b · min(-1)) volunteered to perform the YoYo intermittent recovery test and 6 bouts of soccer drills including 1-a-side, 2-a-side, 3-a-side, and 4-a-side games without a goalkeeper in random order at 2-day intervals. The differences in La-, HR and%HRmax either among the SSGs or among the bouts were identified using 4 x 6 (games x exercise bouts) 2-way analysis of variance with repeated measures. Significant differences were found on La-, HR, and %HRmax among the bouts (p ≤ 0.05). The 3-a-side and 4-a-side games were significantly higher than 1-a-side and 2-a-side games on HR and %HRmax (p ≤ 0.05), whereas the 1-a-side game significantly resulted in higher La- responses compared to other SSGs. This study demonstrated that physiological responses during the 1-a-side and 2-a-side games were different compared to 3-a-side and 4-a-side games. Therefore, it can be concluded that a decreased number of players results in increased intensity during SSGs including 6 bouts. These results suggest that coaches should pay attention on choosing the SSG type and the number of bouts to improve desired physical conditioning of elite young soccer players in soccer training.     

Chronic Intermittent Hypoxia Is Independently Associated with Reduced Postoperative Opioid Consumption in Bariatric Patients Suffering from Sleep-Disordered Breathing

BackgroundEvidence suggests that recurrent nocturnal hypoxemia may affect pain response and/or the sensitivity to opioid analgesia. We tested the hypothesis that nocturnal hypoxemia, quantified by sleep time spent at an arterial saturation (SaO₂) < 90% and minimum nocturnal SaO₂ on polysomnography, are associated with decreased pain and reduced opioid consumption during the initial 72 postoperative hours in patients having laparoscopic bariatric surgery.MethodsWith Institutional Review Board approval, we examined the records of all patients who underwent laparoscopic bariatric surgery between 2004 and 2010 and had an available nocturnal polysomnography study. We assessed the relationships between the time-weighted average of pain score and total opioid consumption during the initial 72 postoperative hours, and: (a) the percentage of total sleep time spent at SaO₂ < 90%, (b) the minimum nocturnal SaO₂, and (c) the number of apnea/hypopnea episodes per hour of sleep. We used multivariable regression models to adjust for both clinical and sleep-related confounders.ResultsTwo hundred eighteen patients were included in the analysis. Percentage of total sleep time spent at SaO₂ < 90% was inversely associated with total postoperative opioid consumption; a 5-%- absolute increase in the former would relatively decrease median opioid consumption by 16% (98.75% CI: 2% to 28%, P = 0.006). However, the percentage of total sleep time spent at SaO₂ < 90% was not associated with pain. The minimum nocturnal SaO₂ was associated neither with total postoperative opioid consumption nor with pain. In addition, neither pain nor total opioid consumption was significantly associated with the number of apnea/hypopnea episodes per hour of sleep.ConclusionsPreoperative nocturnal intermittent hypoxia may enhance sensitivity to opioids.     

Hypoxia induces oxidative stress in tissues of a goby, the rotan Perccottus glenii

The effects of hypoxia (0.4 mg O₂/L) for 2, 6 or 10 h and subsequent normoxic recovery on the levels of lipid peroxides, thiobarbituric acid reactive substances, protein carbonyls (CP), free thiols, and the activities of six antioxidant and associated enzymes were measured in the brain, liver, and skeletal muscle of the rotan Perccottus glenii. Hypoxia increased CP content in the brain (5.0–7.4-fold), liver (2.2–3.3-fold) and muscle (3.2–61-fold) relative to controls and the levels remained elevated during recovery. Lipid peroxide content rose within 2 h of hypoxia in all tissues examined with the most marked increase (8.7-fold) in the liver, but decreased again during longer hypoxic exposure except in the muscle. Levels of low-molecular mass thiols were transiently lowered after 2 h hypoxia in all tissues, but were higher compared with controls after longer hypoxic exposure and recovery. Hypoxia decreased protein thiol content in the liver and muscle that return to control levels during recovery. Experimental conditions affected enzyme activities in a different manner. Superoxide dismutase activity rose two-fold in the liver of hypoxic fish, and a similar tendency was seen in muscle glutathione-S-transferase. Activities of other enzymes were decreased or unchanged during hypoxia and elevated in some cases during normoxic recovery. Taken together, these data show that hypoxia resulted in the development of oxidative stress and a compensatory changes of antioxidant enzymes in the tissues.     

Physiological responses of small-sided vs. regular games in youth volleyball players

The aim of the present study was to evaluate the effects of two different formats of small-sided games (SSGs; 2 vs. 2 and 3 vs. 3) on physiological parameters in youth volleyball players. Twelve youth volleyball players (17.2 ± 7.44 years, 1.89 ± 0.6 m, 72.83 ± 8.57 kg) completed three different games formats (regular game (RG): 6 vs. 6, SSG3: 3 vs. 3 and SSG2: 2 vs. 2). associated with two pitch dimensions (i.e., 2 vs. 2 and 3 vs. 3 on 18 × 4.5 m; 6 vs. 6 on 18 × 9 m). Each player performed 4 × 5 min SSG with a recovery period of 1 min between bouts. All players were members of the same youth team and played in a professional league. They had at least 6 years of volleyball training and no current injuries. This study was carried out during the competitive period. Heart rate (HR), blood lactate ([La⁻]), and rating of perceived exertion (RPE) were measured. Compared to RG, physiological (i.e., HR and [La⁻]) and RPE responses were significantly higher during SSG3 and SSG2 (all p < 0.05; ƞ_p² = 0.77, ƞ_p² = 0.65, ƞ_p² = 0.30, respectively). Moreover, HR and RPE were significantly higher in SSG2 compared to SSG3. In contrast, no significant differences were observed in [La⁻] between SSG2 and SSG3. These results suggest that the number of players influences the exercise intensity in small-sided volleyball games in youth players. Therefore, coaches could benefit from incorporating SSGs to manipulate the exercise intensity in youth volleyball players.     

Neuromuscular fatigue induced by a 90-minute soccer game modeling

This study aimed to quantify neuromuscular fatigue induced by a soccer game. Eight amateur soccer players (age 20.4 ± 1.3 years, mass 70.4 ± 6.9 kg, and height 174.9 ± 5.2 cm) reproduced a 90-minute soccer game modeling composed of two 45-minute periods separated by a 15-minute rest. Torque of quadriceps and hamstring muscle groups associated with electromyography, sprint speed, and vertical jump height was assessed before, at halftime, and immediately after the modeling. Most physical qualities decreased throughout the game with greater decays at match end than at halftime. Contrarily to quadriceps muscles, hamstring torque impairments were not accompanied by electromyographic activity reductions. Squat jump height was reduced at halftime and game end without any change for countermovement jumps. The sprint speed decrease was associated with stride frequency impairments without any change in amplitude and contact time. We concluded on torque production capacity and specific performance impairments during and after soccer games. Neuromuscular fatigue appeared primarily centrally mediated as attested by the reduced quadriceps muscle activity.     

The effect of graded hypoxia on the embryonic chick heart

Embryonic ventricular function in the chick was measured in response to graded levels of hypoxia. Myocardial contractility, as measured by cinephotoanalysis and expressed as shortening fraction, was significantly depressed after 1 hour of moderate hypoxia (6% O2) and after 5 hours of milder (16% O2 and 11% O2) levels of hypoxia (P less than .05). Microscopy confirmed associated myocyte damage with cell death noted after 5 hours of moderate hypoxic stress. Heart rate change was not related to the severity of hypoxia. The greatest level of tachycardia was noted with conditions of mildest hypoxia (16% O2). The data confirm that cardiac contractility, as measured by shortening fraction, is depressed on exposure to hypoxia, with impairment of function related to the severity of the hypoxic conditions.     

Lethal freeze-dehydration injury of dogwood stem tissue does not change the activation energy of water permeability

The Arrhenius activation energy for water permeability, (ΔE_a,H2O) through stem cortical tissue of red osier dogwood (Cornus sericea L.) was determined after treatments which cause membrane rupture as well as after a lethal slow freeze and subsequent slow rewarming. The latter value was higher than the former, but was indistinguishable from the ΔE_a,H2O found for healthy tissue. It was concluded that membrane permeability to water is not altered during the first 24 to 48 hours after exposure of nonacclimated red osier dogwood to lethal freeze dehydration injury.     

Prooxidant/Antioxidant Balance in Hypoxia: A Cross-Over Study on Normobaric vs. Hypobaric “Live High-Train Low”

“Live High-Train Low” (LHTL) training can alter oxidative status of athletes. This study compared prooxidant/antioxidant balance responses following two LHTL protocols of the same duration and at the same living altitude of 2250 m in either normobaric (NH) or hypobaric (HH) hypoxia. Twenty-four well-trained triathletes underwent the following two 18-day LHTL protocols in a cross-over and randomized manner: Living altitude (P_IO₂ = 111.9 ± 0.6 vs. 111.6 ± 0.6 mmHg in NH and HH, respectively); training “natural” altitude (~1000–1100 m) and training loads were precisely matched between both LHTL protocols. Plasma levels of oxidative stress [advanced oxidation protein products (AOPP) and nitrotyrosine] and antioxidant markers [ferric-reducing antioxidant power (FRAP), superoxide dismutase (SOD) and catalase], NO metabolism end-products (NOx) and uric acid (UA) were determined before (Pre) and after (Post) the LHTL. Cumulative hypoxic exposure was lower during the NH (229 ± 6 hrs.) compared to the HH (310 ± 4 hrs.; P<0.01) protocol. Following the LHTL, the concentration of AOPP decreased (-27%; P<0.01) and nitrotyrosine increased (+67%; P<0.05) in HH only. FRAP was decreased (-27%; P<0.05) after the NH while was SOD and UA were only increased following the HH (SOD: +54%; P<0.01 and UA: +15%; P<0.01). Catalase activity was increased in the NH only (+20%; P<0.05). These data suggest that 18-days of LHTL performed in either NH or HH differentially affect oxidative status of athletes. Higher oxidative stress levels following the HH LHTL might be explained by the higher overall hypoxic dose and different physiological responses between the NH and HH.     

Influence of the arachidonic acid cascade on the in vitro hepatic response to hypoxia

Studies were undertaken to determine the effect of arachidonic acid, the precursor of bisenoic prostanoic acid derivatives, on the response of the isolated, perfused rabbit liver to hypoxia. Two and one half hours of severe hypoxia resulted in significant increases in hepatic vascular perfusion pressure, tissue wet weight, and the rates of cellular loss of lactic dehydrogenase, malic dehydrogenase, and acid phosphatase into the perfusing medium. Hypoxia also increased the rate of hepatic PGF_2α production by 25% after 2 hours (p<0.05, hypoxia vs sham). The addition of arachidonic acid (0.1 μg/g/min for 150 minutes) to the perfusion medium of hypoxic livers significantly attenuated the changes in perfusion pressure, tissue wet weight, and loss of cellular enzymes. Arachidonic acid administration increased the rate of PGF_2α production by 100% (p<0.05, sham vs hypoxia + arachidonic acid) within 30 min after hypoxia and maintained this rate for the duration of the study. These results demonstrate that hypoxia mediated prostaglandin F_2α synthesis in the rabbit liver can occur in the absence of neural and blood borne components and that significant activation of the arachidonic acid cascade via the administration of exogenous arachidonic acid has a salutary effect on hepatic hemodynamics and cellular integrity during hypoxia.     

Number of Players and Relative Pitch Area per Player: Comparing Their Influence on Heart Rate and Physical Demands in Under-12 and Under-13 Football Players

The aim of the present study is to analyse the influence of different large-sided games (LSGs) on the physical and physiological variables in under-12s (U12) and -13s (U13) soccer players. The effects of the combination of different number of players per team, 7, 9, and 11 (P7, P9, and P11, respectively) with three relative pitch areas, 100, 200, and 300 m² (A100, A200, and A300, respectively), were analysed in this study. The variables analysed were: 1) global indicator such as total distance (TD); work:rest ratio (W:R); player-load (PL) and maximal speed (V_max); 2) heart rate (HR) mean and time spent in different intensity zones of HR (<75%, 75–84%, 84–90% and >90%), and; 3) five absolute (<8, 8–13, 13–16 and >16 Km h^-1) and three relative speed categories (<40%, 40–60% and >60% V_max). The results support the theory that a change in format (player number and pitch dimensions) affects no similarly in the two players categories. Although it can seem that U13 players are more demanded in this kind of LSG, when the work load is assessed from a relative point of view, great pitch dimensions and/or high number of player per team are involved in the training task to the U12 players. The results of this study could alert to the coaches to avoid some types of LSGs for the U12 players such as: P11 played in A100, A200 or A300, P9 played in A200 or A300 and P7 played in A300 due to that U13>U12 in several physical and physiological variables (W:R, time spent in 84–90%HR_max, distance in 8–13 and 13–16 Km h^-1 and time spent in 40–60%V_max). These results may help youth soccer coaches to plan the progressive introduction of LSGs so that task demands are adapted to the physiological and physical development of participants.     

Effects of Small-Sided Games vs. Interval Training in Aerobic Fitness and Physical Enjoyment in Young Elite Soccer Players

The purpose of this study was to compare the effects of Small-Sided Games (SSG) vs. Interval Training (IT) in soccer training on aerobic fitness and physical enjoyment in youth elite soccer players during the last 8 weeks of the season. Seventeen U-16 male soccer players (age = 15.5 ± 0.6 years, and 8.5 years of experience) of a Spanish First Division club academy were randomized to 2 different groups for 6 weeks: SSG group (n = 9) and IT group (n = 8). In addition to the usual technical and tactical sessions and competitive games, the SSG group performed 11 sessions with different SSGs, whereas the IT group performed the same number of sessions of IT. Players were tested before and after the 6-week training intervention with a continuous maximal multistage running field test and the counter movement jump test (CMJ). At the end of the study, players answered the physical activity enjoyment scale (PACES). During the study, heart rate (HR) and session perceived effort (sRPE) were assessed. SSGs were as effective as IT in maintaining the aerobic fitness in elite young soccer players during the last weeks of the season. Players in the SSG group declared a greater physical enjoyment than IT (P = 0.006; ES = 1.86 ± 1.07). Coaches could use SSG training during the last weeks of the season as an option without fear of losing aerobic fitness while promoting high physical enjoyment.     

DIFFERENT ENDURANCE CHARACTERISTICS OF FEMALE AND MALE GERMAN SOCCER PLAYERS

The aims of the present study were to assess gender differences regarding lactate threshold and intermittent shuttle run performance in female and male soccer players as well as to investigate the relationships between both endurance characteristics in both genders. Fourteen female (1^st division) and thirteen male (4^th division) soccer players completed an incremental test (IT) to determine running velocities at 2 and 4 mmol · l⁻¹ blood lactate (v2 and v4) and maximum velocity (v_max) as well as an interval shuttle run test (ISRT) to determine running distance. Based on v2 and v4 and their percentages in relation to v_max, three intensity zones were calculated: a low lactate zone (<v2), a lactate accommodation zone (v2 to v4), and a lactate accumulation zone (>v4). Female soccer players have a lower v4 (8.2%), v_max (11.3%) and ISRT distance (31.6%). No gender difference was found in v2. In contrast to males, ISRT distance correlates with v_max as well as with v2 and v4 in female soccer players. The intensity zones <v2 and >v4 differ between genders. The present study revealed that gender differences increase when the running performance is intermittent including change of directions. In both genders, different relationships between lactate threshold and intermittent shuttle run performance exist. During incremental testing, the running performances of female and male players reflect different distributions of aerobic and anaerobic metabolic pathways. The revealed gender differences should be considered for soccer endurance training.