New Role for Cdc14 Phosphatase: Localization to Basal Bodies in the Oomycete Phytophthora and Its Evolutionary Coinheritance with Eukaryotic Flagella

Cdc14 protein phosphatases are well known for regulating the eukaryotic cell cycle, particularly during mitosis. Here we reveal a distinctly new role for Cdc14 based on studies of the microbial eukaryote Phytophthora infestans, the Irish potato famine agent. While Cdc14 is transcribed constitutively in yeast and animal cells, the P. infestans ortholog is expressed exclusively in spore stages of the life cycle and not in vegetative hyphae where the bulk of mitosis takes place. PiCdc14 expression is first detected in nuclei at sporulation, and during zoospore formation the protein accumulates at the basal body, which is the site from which flagella develop. The association of PiCdc14 with basal bodies was supported by co-localization studies with the DIP13 basal body protein and flagellar β-tubulin, and by demonstrating the enrichment of PiCdc14 in purified flagella-basal body complexes. Overexpressing PiCdc14 did not cause defects in growth or mitosis in hyphae, but interfered with cytoplasmic partitioning during zoosporogenesis. This cytokinetic defect might relate to its ability to bind microtubules, which was shown using an in vitro cosedimentation assay. The use of gene silencing to reveal the precise function of PiCdc14 in flagella is not possible since we showed previously that silencing prevents the formation of the precursor stage, sporangia. Nevertheless, the association of Cdc14 with flagella and basal bodies is consistent with their phylogenetic distribution in eukaryotes, as species that lack the ability to produce flagella generally also lack Cdc14. An ancestral role of Cdc14 in the flagellar stage of eukaryotes is thereby proposed

Physiological Responses and Physical Performance during Football in the Heat

PURPOSE: To examine the impact of hot ambient conditions on physical performance and physiological responses during football match-play. METHODS: Two experimental games were completed in temperate (∼ 21°C; CON) and hot ambient conditions (∼ 43°C; HOT). Physical performance was assessed by match analysis in 17 male elite players during the games and a repeated sprint test was conducted after the two game trials. Core and muscle temperature were measured and blood samples were obtained, before and after the games. RESULTS: Muscle and core temperatures were ∼ 1°C higher (P<0.05) in HOT (40.3 ± 0.1 and 39.5 ± 0.1°C, respectively) compared to CON (39.2 ± 0.1 and 38.3 ± 0.1°C). Average heart rate, plasma lactate concentration, body weight loss as well as post-game sprint performance were similar between the two conditions. Total game distance declined (P<0.05) by 7% and high intensity running (>14 km ⋅ h(-1)) by 26% in HOT compared to CON), but peak sprint speed was 4% higher (P<0.05) in HOT than in CON, while there were no differences in the quantity or length of sprints (>24 km ⋅ h(-1)) between CON and HOT. In HOT, success rates for passes and crosses were 8 and 9% higher (P<0.05), respectively, compared to CON. Delta increase in core temperature and absolute core temperature in HOT were correlated to total game distance in the heat (r = 0.85 and r = 0.53, respectively; P<0.05), whereas, total and high intensity distance deficit between CON and HOT were not correlated to absolute or delta changes in muscle or core temperature. CONCLUSION: Total game distance and especially high intensity running were lower during a football game in the heat, but these changes were not directly related to the absolute or relative changes in core or muscle temperature. However, peak sprinting speed and execution of successful passes and crosses were improved in the HOT condition

Associations of exercise-induced hormone profiles and gains in strength and hypertrophy in a large cohort after weight training

The purpose of this study was to investigate associations between acute exercise-induced hormone responses and adaptations to high intensity resistance training in a large cohort (n = 56) of young men. Acute post-exercise serum growth hormone (GH), free testosterone (fT), insulin-like growth factor (IGF-1) and cortisol responses were determined following an acute intense leg resistance exercise routine at the midpoint of a 12-week resistance exercise training study. Acute hormonal responses were correlated with gains in lean body mass (LBM), muscle fibre cross-sectional area (CSA) and leg press strength. There were no significant correlations between the exercise-induced elevations (area under the curve—AUC) of GH, fT and IGF-1 and gains in LBM or leg press strength. Significant correlations were found for cortisol, usually assumed to be a hormone indicative of catabolic drive, AUC with change in LBM (r = 0.29, P < 0.05) and type II fibre CSA (r = 0.35, P < 0.01) as well as GH AUC and gain in fibre area (type I: r = 0.36, P = 0.006; type II: r = 0.28, P = 0.04, but not lean mass). No correlations with strength were observed. We report that the acute exercise-induced systemic hormonal responses of cortisol and GH are weakly correlated with resistance training-induced changes in fibre CSA and LBM (cortisol only), but not with changes in strength

Analysis of Dehydration and Strength in Elite Badminton Players

Background: The negative effects of dehydration on aerobic activities are well established. However, it is unknown how dehydration affects intermittent sports performance. The purpose of this study was to identify the level of dehydration in elite badminton players and its relation to muscle strength and power production. Methodology: Seventy matches from the National Spanish badminton championship were analyzed (46 men?s singles and 24 women?s singles). Before and after each match, jump height and power production were determined during a countermovement jump on a force platform. Participants? body weight and a urine sample were also obtained before and after each match. The amount of liquid that the players drank during the match was also calculated by weighing their individual drinking bottles. Results and Discussion: Sweat rate during the game was 1.1460.46 l/h in men and 1.0260.64 l/h in women. The players rehydrated at a rate of 1.1060.55 l/h and 1.0160.44 l/h in the male and female groups respectively. Thus, the dehydration attained during the game was only 0.3760.50% in men and 0.3260.83% in women. No differences were found in any of the parameters analyzed during the vertical jump (men: from 31.8265.29 to 32.9064.49 W/kg; p.0.05, women: from 26.3664.73 to 27.2564.44 W/kg; p.0.05). Post-exercise urine samples revealed proteinuria (60.9% of cases in men and 66.7% in women), leukocyturia (men = 43.5% and women = 50.0%) and erythrocyturia (men = 50.0% and women = 21.7%). Conclusions: Despite a moderate sweat rate, badminton players adequately hydrated during a game and thus the dehydration attained was low. The badminton match did not cause muscle fatigue but it significantly increased the prevalence of proteinuria, leukocyturia and erythrocyturia

Examination of the efficacy of acute L-alanyl-L-glutamine ingestion during hydration stress in endurance exercise

<p>Abstract</p> <p>Background</p> <p>The effect of acute L-alanyl-L-glutamine (AG; Sustamine™) ingestion on performance changes and markers of fluid regulation, immune, inflammatory, oxidative stress, and recovery was examined in response to exhaustive endurance exercise, during and in the absence of dehydration.</p> <p>Methods</p> <p>Ten physically active males (20.8 ± 0.6 y; 176.8 ± 7.2 cm; 77.4 ± 10.5 kg; 12.3 ± 4.6% body fat) volunteered to participate in this study. During the first visit (T1) subjects reported to the laboratory in a euhydrated state to provide a baseline (BL) blood draw and perform a maximal exercise test. In the four subsequent randomly ordered trials, subjects dehydrated to -2.5% of their baseline body mass. For T2, subjects achieved their goal weight and were not rehydrated. During T3 - T5, subjects reached their goal weight and then rehydrated to 1.5% of their baseline body mass by drinking either water (T3) or two different doses (T4 and T5) of the AG supplement (0.05 g·kg^-1and 0.2 g·kg^-1, respectively). Subjects then exercised at a workload that elicited 75% of their VO₂max on a cycle ergometer. During T2 - T5 blood draws occurred once goal body mass was achieved (DHY), immediately prior to the exercise stress (RHY), and immediately following the exercise protocol (IP). Resting 24 hour (24P) blood samples were also obtained. Blood samples were analyzed for glutamine, potassium, sodium, aldosterone, arginine vasopressin (AVP), C-reactive protein (CRP), interleukin-6 (IL-6), malondialdehyde (MDA), testosterone, cortisol, ACTH, growth hormone and creatine kinase. Statistical evaluation of performance, hormonal and biochemical changes was accomplished using a repeated measures analysis of variance.</p> <p>Results</p> <p>Glutamine concentrations for T5 were significantly higher at RHY and IP than T2 - T4. When examining performance changes (difference between T2 - T5 and T1), significantly greater times to exhaustion occurred during T4 (130.2 ± 340.2 sec) and T5 (157.4 ± 263.1 sec) compared to T2 (455.6 ± 245.0 sec). Plasma sodium concentrations were greater (p < 0.05) at RHY and IP for T2 than all other trials. Aldosterone concentrations at RHY and IP were significantly lower than that at BL and DHY. AVP was significantly elevated at DHY, RHY and IP compared to BL measures. No significant differences were observed between trials in CRP, IL-6, MDA, or in any of the other hormonal or biochemical measures.</p> <p>Conclusion</p> <p>Results demonstrate that AG supplementation provided a significant ergogenic benefit by increasing time to exhaustion during a mild hydration stress. This ergogenic effect was likely mediated by an enhanced fluid and electrolyte uptake.</p

The Transcriptome of Compatible and Incompatible Interactions of Potato (Solanum tuberosum) with Phytophthora infestans Revealed by DeepSAGE Analysis

Late blight, caused by the oomycete Phytophthora infestans, is the most important disease of potato (Solanum tuberosum). Understanding the molecular basis of resistance and susceptibility to late blight is therefore highly relevant for developing resistant cultivars, either by marker-assissted selection or by transgenic approaches. Specific P. infestans races having the Avr1 effector gene trigger a hypersensitive resistance response in potato plants carrying the R1 resistance gene (incompatible interaction) and cause disease in plants lacking R1 (compatible interaction). The transcriptomes of the compatible and incompatible interaction were captured by DeepSAGE analysis of 44 biological samples comprising five genotypes, differing only by the presence or absence of the R1 transgene, three infection time points and three biological replicates. 30.859 unique 21 base pair sequence tags were obtained, one third of which did not match any known potato transcript sequence. Two third of the tags were expressed at low frequency (<10 tag counts/million). 20.470 unitags matched to approximately twelve thousand potato transcribed genes. Tag frequencies were compared between compatible and incompatible interactions over the infection time course and between compatible and incompatible genotypes. Transcriptional changes were more numerous in compatible than in incompatible interactions. In contrast to incompatible interactions, transcriptional changes in the compatible interaction were observed predominantly for multigene families encoding defense response genes and genes functional in photosynthesis and CO2 fixation. Numerous transcriptional differences were also observed between near isogenic genotypes prior to infection with P. infestans. Our DeepSAGE transcriptome analysis uncovered novel candidate genes for plant host pathogen interactions, examples of which are discussed with respect to possible function

Analyses of genome architecture and gene expression reveal novel candidate virulence factors in the secretome of Phytophthora infestans

<p>Abstract</p> <p>Background</p> <p><it>Phytophthora infestans </it>is the most devastating pathogen of potato and a model organism for the oomycetes. It exhibits high evolutionary potential and rapidly adapts to host plants. The <it>P. infestans </it>genome experienced a repeat-driven expansion relative to the genomes of <it>Phytophthora sojae </it>and <it>Phytophthora ramorum </it>and shows a discontinuous distribution of gene density. Effector genes, such as members of the RXLR and Crinkler (CRN) families, localize to expanded, repeat-rich and gene-sparse regions of the genome. This distinct genomic environment is thought to contribute to genome plasticity and host adaptation.</p> <p>Results</p> <p>We used <it>in silico </it>approaches to predict and describe the repertoire of <it>P. infestans </it>secreted proteins (the secretome). We defined the "plastic secretome" as a subset of the genome that (i) encodes predicted secreted proteins, (ii) is excluded from genome segments orthologous to the <it>P. sojae </it>and <it>P. ramorum </it>genomes and (iii) is encoded by genes residing in gene sparse regions of <it>P. infestans </it>genome. Although including only ~3% <it>of P. infestans </it>genes, the plastic secretome contains ~62% of known effector genes and shows >2 fold enrichment in genes induced <it>in planta</it>. We highlight 19 plastic secretome genes induced <it>in planta </it>but distinct from previously described effectors. This list includes a trypsin-like serine protease, secreted oxidoreductases, small cysteine-rich proteins and repeat containing proteins that we propose to be novel candidate virulence factors.</p> <p>Conclusions</p> <p>This work revealed a remarkably diverse plastic secretome. It illustrates the value of combining genome architecture with comparative genomics to identify novel candidate virulence factors from pathogen genomes.</p

EFFECTS OF INCREASING AMBIENT HUMIDITY DURING LOW-INTENSITY EXERCISE IN THE HEAT ON FEMALES

Nicole E. Moyen1,2,3, Toby Mündel2, Andrea M. Du Bois1, Anthony B. Ciccone1, and Daniel A. Judelson1, FACSM; 1Exercise Physiology Laboratory, Department of Kinesiology, California State University, Fullerton; 2 School of Sport and Exercise, Massey University, Palmerston North, New Zealand; 3Human Performance Laboratory, Department of Health, Human Performance, and Recreation, University of Arkansas Females increasingly occupy vocations that require working prolonged periods in hot-humid climates, however, research examining females under these conditions remains unclear. PURPOSE: To assess how increasing relative humidity (RH) influences females’ thermoregulation and perception during low-intensity exercise in the heat. METHODS: Ten healthy females (age=23±2 y, VO2max=46±6 ml×kg-1×min-1) walked 90 min at 35%VO2max in 35˚C at 55% RH (55RH), 70% RH (70RH), and 85% RH (85RH); all trials were completed during follicular phase. Investigators obtained 1) rectal temperature (Tre), mean-weighted skin temperature (Tsk), heart rate (HR), ambient temperature (Ta), and RH every five min, 2) pre and post nude body masses, and 3) respiratory measures every 30 min; partitional calorimetric variables were calculated from these measures. RESULTS: Significant RH*time interactions existed for all calorimetry variables (n=9; all p\u3c0.0005) except heat production (p=0.139), convective respiratory heat loss (p=0.137), and required rate of evaporative cooling (Ereq; p=0.197). Each RH increase significantly decreased evaporative heat loss (E); all other calorimetry variables only significantly differed after 70RH. Combined, dry (convective+radiative) and respiratory heat losses only compensated for \u3c30% of decreases in E. Tre, Tsk, and HR presented significant RH*time interactions (p\u3c0.0005). 85RH Tre significantly exceeded 55RH and 70RH after 35 min. 70RH Tsk tended to exceed 55RH (p=0.057), and 85RH Tsk significantly exceeded 55RH and 70RH after 30 and 40 min, respectively. After 10 min, 85RH HR significantly exceeded 55RH and 70RH; after 50 min, 70RH HR significantly exceeded 55RH. 70RH sweat rate tended to exceed 55RH (p=0.063), and 85RH significantly exceeded both trials (55RH=0.6±0.2; 70RH=0.7±0.2; 85RH=0.9±0.3 L×h-1; p=0.001). CONCLUSION: The primary findings of this study were that females exercising at low intensities in 35°C only experienced significant thermoregulatory strain after RH exceeded 70%. As heat production and Ereq remained constant across trials, heat storage increased with each 15% rise in RH because dry and respiratory heat losses minimally offset decreased E. Hotter Tre,Tsk, and resultantly higher sweat rate, reflected heat storage increases as E decreased in each trial. Overall, at Ta ≤ 35 °C, we found females working/exercising for prolonged periods at low-intensities will remain at safe core temperatures up to 70% RH

Influence of diuretic-induced dehydration on competitive sprint and power performance

Purpose: To examine sprint and power performance after diuretic-induced dehydration. Methods: After six sprint practice sessions, nine male former sprinters (mean ± SD; age, 21 ± 2 yr; body mass (BM), 80.0 ± 5.2 kg; height, 1.78 ± 0.08 m; body fat, 14 ± 4%) participated in a 50-m race, a 200-m race, a 400-m race, and a vertical jump on an indoor synthetic track, once when dehydrated (40-mg furosemide; DD) and once with no diuretic (CON) using a counter-balanced crossover design. Plasma volume change (%ΔPV), heart rate (HR), blood pressure, rectal temperature, serum electrolytes, plasma lactate, plasma glucose, rating of perceived exertion, thirst, and thermal sensations were measured before and after each race. Results: Sprint times (DD vs CON) for the 50 m (6.72 ± 0.28 vs 6.73 ± 0.29 s), 200 m (25.95 ± 1.20 vs 26.21 ± 1.42 s), and 400 m (59.01 ± 4.26 vs 58.68 ± 3.68 s) were similar for both conditions, as was vertical jump height (0.67 ± 0.10 vs 0.66 ± 0.11 m). This occurred despite losing 2.2 ± 0.4% BM and 7.3 ± 6.7%ΔPV (50/200 m) or 2.5 ± 0.4% BM and 7.1 ± 2.7% %ΔPV (VJ/400 m) in response to DD. Conclusions: Diuretic-induced dehydration was not detrimental to sprint and power performance. Metabolic, thermoregulatory, and cardiovascular variables were not significantly altered by DD. Furthermore, the theoretical benefit of dehydration on performance (i.e., BM reduction) was not supported in this subject cohort

The effects of progressive dehydration on strength and power: is there a dose response?

This study examined the effect of exercise- and heat-induced dehydration on strength, jump capacity and neuromuscular function. Twelve recreationally active males completed six resistance exercise bouts (baseline and after each 5 exposure sessions) in an increasing state of hypohydration obtained by repeated heat exposure and exercise sessions (5 periods of 20 min jogging at up to ~80% age predicted heart rate maximum at 48.5 +/- 0.48 degrees C, relative humidity 50 +/- 4%). Relative to starting values, body mass decreased 1.0 +/- 0.5, 1.9 +/- 0.7, 2.6 +/- 0.8, 3.3 +/- 0.9 and 3.9 +/- 1.0% after exposure 1, 2, 3, 4 and 5, respectively. However, plasma volume remained constant. No significant differences existed amongst trials in vertical jump height, electromyography data or isokinetic leg extension at a rate of 120 degrees s(-1). Isometric leg extensions were significantly reduced (P < 0.05) after the first (1% body mass loss) and subsequent exposures in comparison to baseline. Isokinetic leg extensions at a rate of 30 degrees s(-1) were significantly reduced after the third (2.6% body mass loss) and subsequent exposures compared with baseline. No dose response was identified in any of the tested variables yet a threshold was observed in isometric and isokinetic strength at 30 degrees s(-1). In conclusion, dehydration caused by jogging in the heat had no effect on vertical jumping or isokinetic leg extensions at a rate of 120 degrees s(-1). Alternatively, exercise-induced dehydration was detrimental to isometric and isokinetic leg extensions at a rate of 30 degrees s(-1), suggesting the force-velocity relationship in hypohydration merits further research