Movement demands in Australian rules football as indicators of muscle damage

The purpose of this study was to determine if there is an association between variables that describe movements in an Australian Rules football (ARF) game with muscle damage. Fourteen elite junior ARF players were monitored with a global positioning system (GPS) during a match, and muscle damage was estimated by determining creatine kinase (CK) 24 hours postmatch. The players were median split based on CK levels, into a high and low CK group, and the groups were compared with independent t-tests. The primary finding was that the group that experienced greater muscle damage (high CK group) generally covered significantly (p < 0.05) greater distances. This was the case for running speeds between 4 and 7 m·s(-1) and, with the exception of high acceleration, all intensities of acceleration and deceleration. The high, as compared with the low, CK group also produced a significantly greater (42%) "player load." All of these significant differences were accompanied by large effect sizes. Group-specific Pearson (r) correlations between CK level and GPS variables suggest that a certain volume of movement is required before the elicitation of a positive relationship beyond trivial or small. Correlations between CK and running speeds >4 m·s(-1) and moderate-high acceleration and deceleration were negative in the low CK (lesser volumes) group. With the exception of low-intensity acceleration/deceleration, the same relationships were positive and generally of a moderate-to-large magnitude in the high CK (greater volumes) group. It may be that a certain volume of movement is required for that movement to be strongly associated with CK levels. It was concluded that selected GPS variables obtained from ARF games can be used as indicators of muscle damage, and this information may be used to individualize recovery strategies after games.     

Significance of Ubiquitin Carboxy-Terminal Hydrolase L1 Elevations in Athletes after Sub-Concussive Head Hits

The impact of sub-concussive head hits (sub-CHIs) has been recently investigated in American football players, a population at risk for varying degrees of post-traumatic sequelae. Results show how sub-CHIs in athletes translate in serum as the appearance of reporters of blood-brain barrier disruption (BBBD), how the number and severity of sub-CHIs correlate with elevations of putative markers of brain injury is unknown. Serum brain injury markers such as UCH-L1 depend on BBBD. We investigated the effects of sub-CHIs in collegiate football players on markers of BBBD, markers of cerebrospinal fluid leakage (serum beta 2-transferrin) and markers of brain damage. Emergency room patients admitted for a clinically-diagnosed mild traumatic brain injury (mTBI) were used as positive controls. Healthy volunteers were used as negative controls. Specifically this study was designed to determine the use of UCH-L1 as an aid in the diagnosis of sub-concussive head injury in athletes. The extent and intensity of head impacts and serum values of S100B, UCH-L1, and beta-2 transferrin were measured pre- and post-game from 15 college football players who did not experience a concussion after a game. S100B was elevated in players experiencing the most sub-CHIs; UCH-L1 levels were also elevated but did not correlate with S100B or sub-CHIs. Beta-2 transferrin levels remained unchanged. No correlation between UCH-L1 levels and mTBI were measured in patients. Low levels of S100B were able to rule out mTBI and high S100B levels correlated with TBI severity. UCH-L1 did not display any interpretable change in football players or in individuals with mild TBI. The significance of UCH-L1 changes in sub-concussions or mTBI needs to be further elucidated.     

Circannual rhythm of plasmatic vitamin D levels and the association with markers of psychophysical stress in a cohort of Italian professional soccer players

Adequate plasmatic Vitamin D levels are crucial to maintain calcium homeostasis and bone metabolism both in the general population and in athletes. Correct dietary supply and a regular sun exposure are fundamental for allowing the desired and effective fitness level. Past studies highlighted a scenario of Vitamin D insufficiency among professional soccer players in several countries, especially in North Europe, whilst a real deficiency in athletes is rare. The typical seasonal fluctuations of Vitamin D are wrongly described transversally in athletes belonging to teams that play at different latitudes and a chronobiologic approach studying the Vitamin D circannual rhythm in soccer players has not been described yet. Therefore, we studied plasma vitamin D, cortisol, testosterone, and creatin kinase (CK) concentrations in three different Italian professional teams training at the same latitude during a period of two consecutive competitive seasons (2013 and 2014). In this retrospective observational study, 167 professional soccer players were recruited (mean age at sampling 25.1 ± 4.7 years) and a total of 667 blood drawings were carried out to determine plasma 25(OH)D, serum cortisol, serum testosterone and CK levels. Testosterone to cortisol ratio (TC) was calculated based as a surrogate marker of overtraining and psychophysical stress and each athlete was drawn until a maximum of 5 times per season. Data extracted by a subgroup of players that underwent at least 4 sample drawings along a year (N = 45) were processed with the single and population mean cosinor tests to evaluate the presence of circannual rhythms: the amplitude (A), acrophase (Φ) and the MESOR (M) are described. In total, 55 players (32.9%) had an insufficient level of 25(OH)D during the seasons and other 15 athletes (9.0%) showed, at least once, a deficiency status of Vitamin D. The rhythmometric analyses applied to the data of Vitamin D revealed the presence of a significant circannual rhythm (p < 0.001) with the acrophase that occurred in August; the rhythms of Vitamin D levels were not different neither among the three soccer teams nor between competitive seasons. Cortisol, testosterone and TC showed significant circannual rhythms (p < 0.001): cortisol registered an acrophase during winter (February) while testosterone and TC registered their peaks in the summer months (July). On the contrary, CK did not display any seasonal fluctuations. In addition, we observed weak but significant correlations between 25(OH)D versus testosterone (r = 0.29 and p < 0.001), cortisol (r = ?0.27 and p < 0.001) and TC (r = 0.37 and p < 0.001). No correlation was detected between Vitamin D and CK. In conclusion, the correct chronobiologic approach in the study of annual variations of Vitamin D, cortisol and testosterone could be decisive in the development of more specific supplementation and injury prevention strategies by athletic trainers and physicians.     

Inflammatory response and neutrophil functions in players after a futsal match

Inflammatory response and neutrophil functions in players after a futsal match. J Strength Cond Res 26(9): 2507-2514, 2012-Futsal players suffer injuries resulting from muscle fatigue and contact or collision among players. Muscle lesions can be detected by measuring muscle lesion markers such as creatine kinase (CK) and lactate dehydrogenase (LDH) in plasma. After an initial lesion, there is an increase in the plasma levels of C-reactive protein (CRP) and proinflammatory cytokines. These mediators may activate neutrophils and contribute to tissue damage and increase susceptibility to invasive microorganisms. In this study, we investigated the effect of a futsal match on muscle lesion markers, cytokines, and CRP in elite players. The basal and stimulated neutrophil responsiveness after a match was also evaluated based on measurements of neutrophil necrosis, apoptosis, phagocytic capacity, reactive oxygen species (ROS) production, and cytokines (tumor necrosis factor-alpha [TNF-α], interleukin [IL]-8, IL-1β, IL-10, and IL-1ra) production. Blood samples were taken from 16 players (26.4 ± 3.2 years, 70.2 ± 6.9 kg, 59.7 ± 5.1 ml·kg·min, sports experience of 4.4 ± 0.9 years) before and immediately after a match. Exercise increased the serum activities of CK (2.5-fold) and LDH (1.3-fold). Playing futsal also increased the serum concentrations of IL-6 (1.6-fold) and CRP (1.6-fold). The TNF-α, IL-1β, IL-8, IL-1ra, and IL-10 serum levels were not modified in the conditions studied. The futsal match induced neutrophil apoptosis, as indicated by phosphatidylserine externalization (6.0-fold). The exercise induced priming of neutrophils by increasing ROS (1.3-fold), TNF-α (5.8-fold), and IL-1β (4.8-fold) released in nonstimulated cells. However, in the stimulated condition, the exercise decreased neutrophil function, diminishing the release of ROS by phorbol myristate acetate-stimulated neutrophils (1.5-fold), and the phagocytic capacity (1.6-fold). We concluded that playing futsal induces inflammation, primes and activates neutrophils, and reduces the efficiency of neutrophil phagocytosis immediately after a match.     

Effects of handling stress on plasma enzymes in harp seals, Phoca groenlandica.

Three harp seal pups, Phoca groenlandica, were captured on the ice of the Gulf of St. Lawrence, and subjected to 3 h of transportation and handling stress. The activities of creatine kinase (CK), aspartate aminotransferase (AspAT), aldolase, alanine aminotransferase, gamma glutamyl transpeptidase, and leucine aminopeptidase were determined in serial blood samples collected for 4 d following the stress episode. Marked elevation of plasma CK activity was observed 3 h after capture. Values returned to normal in 12 h in two seals, and by 24 h in the third. Slight elevations in AspAT were also noted; the remaining enzymes were unaffected. Plasma CK is recommended as a sensitive indicator of handling stress in seals.     

Plyometric training effects on physical fitness and muscle damage in high school baseball players

[Purpose]The objective of this study was to investigate the effects of plyometric training on physical fitness and muscle damage in high school baseball players.[Methods]The participants in the study included 21 high school baseball players who are healthy and well-training. The participants were randomly allocated to the plyometric training (n=11) and control (n=10) groups. The plyometric training was applied 3 times a week for a total of 8 weeks and the control group took part in only regular baseball skills training without plyometric training. For physical fitness, measures included maximal strength (left and right hand-grip strength), muscle endurance (sit-up), agility (side-step), power (standing long jump), and balance (left and right Rhomberg test). For muscle damage, creatine kinase (CK) and lactate dehydrogenase (LDH) levels were measured.[Results]The results showed a significant interaction effect between time and groups in left hand-grip strength (P = 0.022), side-step (P = 0.004), and standing long jump (P < 0.001) after the 8-week plyometric training, with greater improvement in the plyometric training group than the control group. On the other hand, there was no significant interaction effect between time and groups in right hand-grip strength, situp, left and right Rhomberg test, CK level, and LDH level (P > 0.05).[Conclusion]In conclusion, 8-week plyometric training had a positive effect on improving physical fitness, such as maximal strength, agility, and power, in high school baseball players without causing additional muscle damage.     

Effects of High-Intensity Training and Resumed Training on Macroelement and Microelement of Elite Basketball Athletes

The purpose of this study was to assess the effects of high-intensity training and resumed training in hot and humid environment on plasma macro- and microelements levels of elite Han Chinese basketball players. Ten well-trained elite basketball athletes?? plasma macroelements (chlorin, sodium, potassium, and calcium), creatine kinase (CK), and creatine kinase-MB (CK-MB) were measured before and after a 2-h high-intensity training, and microelements (zinc, copper, iron, and selenium) were determined before and after a 1-week high-intensity training and after a 1-week resumed training. The blood CK and CK-MB levels of the elite basketball athletes were significantly increased (P?<?0.05) after high-intensity basketball training. The macroelements (chlorin, sodium, and calcium) levels of blood increased significantly except potassium after high-intensity basketball training. No significant differences (P?>?0.05) were found in zinc and copper levels; nevertheless, the levels of plasma selenium and plasma iron were significantly lower (P?<?0.05) after a 1-week high-intensity training. After a 1-week resumed training, except zinc, all of microelements measured had a trend toward original levels. These results implicated that high-intensity training would provoke the change of macroelements which would lead to electrolyte disturbance. In addition, the present study suggested that a 1-week high-intensity training would have an impact on microelement levels, especially for selenium and iron.     

Deletion of delta-opioid receptor in mice alters skin differentiation and delays wound healing

In addition to their well-known antinociceptive action, opioids can modulate non-neuronal functions, such as immune activity and physiology of different cell types. Several findings suggest that the delta-opioid receptor (DOR) and its endogenous ligands (enkephalins) are important players in cell differentiation and proliferation. Here we show the expression of DOR in mouse skin and human skin cultured fibroblasts and keratinocytes using RT-PCR. In DOR knock-out (KO) mice, a phenotype of thinner epidermis and higher expression of cell differentiation marker cytokeratin 10 (CK 10) were observed compared with wild type (WT). Using a burn wound model, significant wound healing delay (about 2 days) and severe epidermal hypertrophy were shown at the wound margin of DOR KO mice. This wound healing delay was further investigated by immunohistochemistry using markers for proliferation, differentiation, re-epithelialization, and dermal repair (CK 6, CK 10, and collagen IV). The levels of all these markers were increased in wounds of KO mice compared with WT. During the wound healing, the epidermal thickness in KO mice augments faster and exceeds that of the WT by day 3. These results suggest an essential role of DOR in skin differentiation, proliferation, and migration, factors that are important for wound healing.     

Effects of whole-body cryotherapy on serum mediators of inflammation and serum muscle enzymes in athletes

Whole-body cryotherapy (WBC) covers a wide range of therapeutic applications and consists of briefly exposing the body to extremely cold air. In sports medicine, WBC is used to improve recovery from muscle injury; however, empirical studies on its application to this area are lacking. To fill this gap, we compared changes in immunological parameters (C3, IgA, IgM, IgG, C-reactive protein, PGE2), cytokines (IL-2, IL-8, IL-10), adhesion molecules (sICAM-1), and muscle enzymes (creatine kinase [CK], lactate dehydrogenase [LAD]) before and after WBC in 10 top-level Italian National team rugby players. The subjects underwent five sessions on alternate days once daily for 1 week. During the study period, the training workload was the same as that of the previous weeks. Compared to baseline values, immunological parameters remained unchanged, while CK and LAD levels significantly decreased after treatment. No alterations in immunological function were observed but there is a decrease in pro-inflammatory cytokine/chemokine and an increase in anti-inflammatory cytokine.As measured by changes in serum CK and LAD concentrations, and cytokines pathway, short-term cold air exposure was found to improve recovery from exercise-induced muscle injury and/or damage associated with intense physical training.     

Sexually stimulated testosterone release in male mice (Mus musculus): roles of genotype and sexual arousal

In virtually every mammalian species examined, some males exhibit reflexive testosterone release upon encountering a novel female (or female-related stimulus). At the same time, not every individual male (or every published study) provides evidence for reflexive testosterone release. Four experiments using house mice (Mus musculus) examined the hypothesis that both the male's genotype and his degree of sexual arousal (as indexed by ultrasonic mating calls) are related to such variability. In Experiment 1, CF-1 males exhibited reflexive testosterone elevations 30 min after encountering female urine. CK males, on the other hand, did not exhibit testosterone elevations 20, 30, 50, 60, or 80 min after encountering female urine (Experiments 1 and 2) suggesting this strain incapable of reflexive release. In Experiment 3, we measured both mating calls and reflexive testosterone release in response to female urine in CF-1 and CK males. Most males of both strains called vigorously to female urine but not to water. But, only CF-1 males exhibited significant testosterone elevations to female urine. In Experiment 4, DBA/2J males called vigorously to females followed by testosterone elevations 30 min later. The first 3 experiments support the hypothesis that male genotype is an important variable underlying mammalian reflexive testosterone release. Statistically significant correlations between mating calls in the first minute after stimulus exposure and testosterone elevations 30 min later (Experiments 3 and 4) support the hypothesis that, in capable males, reflexive testosterone release is related to the male's initial sexual arousal.     

Biochemical and endocrine responses to impact and collision during elite Rugby League match play

The purpose of this study was to investigate the relationship between the prematch and short-term postmatch biochemical and endocrine responses to the intensity, number, and distribution of impacts associated with collisions during elite Rugby League match play. Seventeen elite male Rugby League players each provided blood and saliva samples 24 hours prematch, 30 minutes prematch, 30 minutes postmatch, and then at 24-hour intervals for a period of 5 days postmatch to determine plasma creatine kinase concentration ([CK]) and salivary cortisol concentration ([sCort]). The intensity, number, and distribution of impact forces experienced by players during match play were recorded using portable global positioning systems (GPSs). The change in the dependent variables at each sample collection time was compared to 24 hours prematch and 30-minute prematch measures. The [CK] and [sCort] increased significantly (p < 0.05) during match play. Significant correlations (p < 0.05) were observed between the number of hit-ups and peak [CK] 24 hours postmatch, 48 hours postmatch, and 72 hours postmatch (p < 0.05). The number of impacts recorded in zone 5 (8.1-10.0G) and zone 6 (>10.1G) during match play was significantly correlated (p < 0.05) to [CK] 30 minutes postmatch, 24 hours post, 48 hours post, and 72 hours postmatch. The GPS was able to provide data on the intensity, number, and distribution of impacts resulting from collisions during match play. Elite Rugby League match play resulted in significant skeletal muscle damage and was highly dependent on the number of heavy collisions >8.1G. [CK] remained elevated 120 hours postmatch identifying that at least 5 days modified activity is required to achieve full recovery after elite Rugby League match play.     

MUSCLE DAMAGE AFTER A TENNIS MATCH IN YOUNG PLAYERS

The present study investigated changes in indirect markers of muscle damage following a simulated tennis match play using nationally ranked young (17.6 ± 1.4 years) male tennis players. Ten young athletes played a 3-hour simulated match play on outdoor red clay courts following the International Tennis Federation rules. Muscle soreness, plasma creatine kinase activity (CK), serum myoglobin concentration (Mb), one repetition maximum (1RM) squat strength, and squat jump (SJ) and counter movement jump (CMJ) heights were assessed before, immediately after, and 24 and 48 h after the simulated match play. All parameters were also evaluated in a non-exercised group (control group). A small increase in the indirect markers of muscle damage (muscle soreness, CK and Mb) was detected at 24-48 hours post-match (p < 0.05). A marked acute decrement in neuromuscular performance (1RM squat strength: -35.2 ± 10.4%, SJ: -7.0 ± 6.0%, CMJ: -10.0 ± 6.3%) was observed immediately post-match (p < 0.05). At 24 h post-match, the 1RM strength and jump heights were not significantly different from the baseline values. However, several players showed a decrease of these measures at 24 h after the match play. The simulated tennis match play induced mild muscle damage in young players. Coaches could monitor changes in the indirect markers of muscle damage to assess athletes’ recovery status during training and competition.     

Interactive effect of an acute bout of resistance exercise and dehydroepiandrosterone administration on glucose tolerance and serum lipids in middle-aged women

The present study determined the interactive effect of an acute bout of resistance exercise and dehydroepiandrosterone (DHEA) administration on glucose tolerance and serum lipids. Twenty middle-aged female subjects performed an acute bout of resistance exercise and were subsequently divided into two groups: placebo (age 40.7 +/- 2.0) and DHEA administered (age 39.0 +/- 2.7). Ten subjects who received DHEA (age 41.5 +/- 4.6) participated in a non-exercise control. DHEA (25 mg twice daily) or placebo was orally supplemented for 48 hours. Before exercise and 48 hours after the last exercise bout (14 hours after the last DHEA intake), an oral glucose tolerance test and an insulin concentration were determined. Levels of fasting serum cholesterol and triglyceride, tumor necrosis factor-alpha (TNF-alpha), creatine kinase (CK) were also measured. The DHEA administration significantly elevated the fasting dehydroepiandrosterone sulfate (DHEA-S) level by approximately 3-fold. Both acute resistance exercise and DHEA administration improved glucose tolerance, but no addictive effect was found. Furthermore, exercise and DHEA administration did not affect serum triglyceride and cholesterol levels, but both lipids were significantly lowered when DHEA was given following exercise. Resistance exercise induced elevations in serum CK and TNFalpha levels, but these increases were attenuated by the DHEA administration. The new finding of this study was that post-exercise DHEA administration decreased serum triglycerides and cholesterol. This effect appeared to be associated with its TNF-alpha lowering action.     

Ocular Delivery of Compacted DNA-Nanoparticles Does Not Elicit Toxicity in the Mouse Retina

Subretinal delivery of polyethylene glycol-substituted lysine peptide (CK30PEG)-compacted DNA nanoparticles results in efficient gene expression in retinal cells. This work evaluates the ocular safety of compacted DNA nanoparticles. CK30PEG-compacted nanoparticles containing an EGFP expression plasmid were subretinally injected in adult mice (1 µl at 0.3, 1.0 and 3.0 µg/µl). Retinas were examined for signs of inflammation at 1, 2, 4 and 7 days post-injection. Neither infiltration of polymorphonuclear neutrophils or lymphocytes was detected in retinas. In addition, elevation of macrophage marker F4/80 or myeloid marker myeloperoxidase was not detected in the injected eyes. The chemokine KC mRNA increased 3–4 fold in eyes injected with either nanoparticles or saline at 1 day post-injection, but returned to control levels at 2 days post-injection. No elevation of KC protein was observed in these mice. The monocyte chemotactic protein-1, increased 3–4 fold at 1 day post-injection for both nanoparticle and saline injected eyes, but also returned to control levels at 2 days. No elevations of tumor necrosis factor alpha mRNA or protein were detected. These investigations show no signs of local inflammatory responses associated with subretinal injection of compacted DNA nanoparticles, indicating that the retina may be a suitable target for clinical nanoparticle-based interventions.     

Altered cytokinin metabolism affects cytokinin, auxin, and abscisic acid contents in leaves and chloroplasts, and chloroplast ultrastructure in transgenic tobacco

Cytokinins (CKs) are involved in the regulation of plant development including plastid differentiation and function. Partial location of CK biosynthetic pathways in plastids suggests the importance of CKs for chloroplast development. The impact of genetically modified CK metabolism on endogenous CK, indole-3-acetic acid, and abscisic acid contents in leaves and isolated intact chloroplasts of Nicotiana tabacum was determined by liquid chromatography/mass spectrometry and two-dimensional high-performance liquid chromatography, and alterations in chloroplast ultrastructure by electron microscopy. Ectopic expression of Sho, a gene encoding a Petunia hybrida isopentenyltransferase, was employed to raise CK levels. The increase in CK levels was lower in chloroplasts than in leaves. CK levels were reduced in leaves of tobacco harbouring a CK oxidase/dehydrogenase gene, AtCKX3. The total CK content also decreased in chloroplasts, but CK phosphate levels were higher than in the wild type. In a transformant overexpressing a maize beta-glucosidase gene, Zm-p60.1, naturally targeted to plastids, a decrease of CK-O-glucosides in chloroplasts was found. In leaves, the changes were not significant. CK-O-glucosides accumulated to very high levels in leaves, but not in chloroplasts, of plants overexpressing a ZOG1 gene, encoding trans-zeatin-O-glucosyltransferase from Phaseolus lunatus. Manipulation of the CK content affected levels of indole-3-acetic and abscisic acid. Chloroplasts of plants constitutively overexpressing Sho displayed ultrastructural alterations including the occasional occurrence of crystalloids and an increased number of plastoglobuli. The other transformants did not exhibit any major differences in chloroplast ultrastructure. The results suggest that plant hormone compartmentation plays an important role in hormone homeostasis and that chloroplasts are rather independent organelles with respect to regulation of CK metabolism.     

Predominance of CK2α over CK2α′ in the mammalian brain

CK2 is a heterotetrameric ubiquitous kinase consisting of two catalytic subunits and two regulatory subunits. The two catalytic subunits, α and α', are highly homologous but differ in their C-terminal regions. It is not known whether CK2α and α' have distinctive substrate specificity, since no α- or α'-specific substrate has been identified. Thus, it is assumed that the two kinase isoforms overlap in their substrate specificity. CK2 protein levels and activity were found to be elevated in the brain when compared to other organs. Here we have studied the protein levels of CK2α and α' isoforms in nine major brain regions. We found that both, CK2α and α', are expressed in all brain regions tested. Whereas CK2α levels do not vary strongly across the regions, CK2α' levels are slightly higher in the cortex and hippocampus than in other regions. Furthermore, we show that CK2α protein levels in the striatum are relatively high when compared to CK2α'. The approximate stoichiometry ratio of CK2α:CK2α' is 8:1. Therefore, one can consider that CK2α levels are predominant in comparison to CK2α' levels throughout the mammalian brain.     

Evaluation of a new radioimmunological assay for the determination of the B subunit of creatine kinase enzyme

We used a new radioimmunological (RIA) kit for the assay of B subunit of creatine kinase enzyme (CK). This RIA system uses a specific antisera against the B subunit as ligant, human CK-BB labelled with 125I as tracer, and purified human CK-BB isoenzyme as standard. The mean (+/- SD) sensitivity obtained was 0.25 +/- 0.16 ng/tube and the between assay variability was about 9-10%. Serum levels of 113 normal subjects was not normally distributed. The 95% of values was found below 5 ng/ml. This new RIA is usefull in clinical practice when serum levels of CK-BB isoenzyme must be determined. This method is quickly and it is characterized by a good degree of precision, but the CK-MB isoenzyme cross-reacts for about 40% in this RIA system. Therefore, for the clinical diagnosis by means of this RIA it is necessary to rule out the concomitant elevations of serum CK-MB values.     

Oxidative modification of creatine kinase BB in Alzheimer's disease brain

Creatine kinase (CK) BB, a member of the CK gene family, is a predominantly cytosolic CK isoform in the brain and plays a key role in regulation of the ATP level in neural cells. CK BB levels are reduced in brain regions affected by neurodegeneration in Alzheimer's disease (AD), Pick's disease, and Lewy body dementia, and this reduction is not a result of decreased mRNA levels. This study demonstrates that posttranslational modification of CK BB plays a role in the decrease of CK activity in AD brain. The specific CK BB activity and protein carbonyl content were determined in brain extracts of six AD and six age-matched control subjects. CK BB activity per microgram of immunoreactive CK BB protein was lower in AD than in control brain extracts, indicating the presence of inactive CK BB molecules. The analysis of specific protein carbonyl levels in CK BB, performed by two-dimensional fingerprinting of oxidatively modified proteins, identified CK BB as one of the targets of protein oxidation in the AD brain. The increase of protein carbonyl content in CK BB provides evidence that oxidative posttranslational modification of CK BB plays a role in the loss of CK BB activity in AD.     

Identification and Expression Analysis of Cytokinin Metabolic Genes in Soybean under Normal and Drought Conditions in Relation to Cytokinin Levels

Cytokinins (CKs) mediate cellular responses to drought stress and targeted control of CK metabolism can be used to develop drought-tolerant plants. Aiming to manipulate CK levels to improve drought tolerance of soybean cultivars through genetic engineering of CK metabolic genes, we surveyed the soybean genome and identified 14 CK biosynthetic (isopentenyltransferase, GmIPT) and 17 CK degradative (CK dehydrogenase, GmCKX) genes. Comparative analyses of GmIPTs and GmCKXs with Arabidopsis counterparts revealed their similar architecture. The average numbers of abiotic stress-inducible cis-elements per promoter were 0.4 and 1.2 for GmIPT and GmCKX genes, respectively, suggesting that upregulation of GmCKXs, thereby reduction of CK levels, maybe the major events under abiotic stresses. Indeed, the expression of 12 GmCKX genes was upregulated by dehydration in R2 roots. Overall, the expressions of soybean CK metabolic genes in various tissues at various stages were highly responsive to drought. CK contents in various organs at the reproductive (R2) stage were also determined under well-watered and drought stress conditions. Although tRNA-type GmIPT genes were highly expressed in soybean, cis-zeatin and its derivatives were found at low concentrations. Moreover, reduction of total CK content in R2 leaves under drought was attributable to the decrease in dihydrozeatin levels, suggesting a role of this molecule in regulating soybean's responses to drought stress. Our systematic analysis of the GmIPT and GmCKX families has provided an insight into CK metabolism in soybean under drought stress and a solid foundation for in-depth characterization and future development of improved drought-tolerant soybean cultivars by manipulation of CK levels via biotechnological approach.     

Does underwater rugby stimulate the over-production of reactive oxygen species?

Antioxidant enzymes in erythrocytes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and antioxidant sex hormone oestradiol in serum and malondialdehyde (MDA) production as a marker of lipid peroxidation in erythrocytes were investigated in male and female underwater rugby (UWR) players. Results showed that except for GSH-Px activity in female players, all antioxidant enzymes increased significantly (p < 0.05) after an UWR game. It was interesting to note that while the concentration of oestradiol in female players did not change, it increased significantly (p < 0.05) in male players. Lipid peroxidation (LPO) levels in female players as well as oestradiol concentration did not change significantly (p > 0.05) whereas LPO levels in male players increased significantly (p < 0.05) compared to pre-exercise values. In conclusion, the results showed that underwater rugby can stimulate over-production of ROS and antioxidant systems and affect oestradiol levels in male players. Because of increased LPO levels observed in male players, complex antioxidant supplementation including co-factors of antioxidant enzymes such as Cu, Zn, Fe, Se and antioxidant vitamins such as vitamin C and E may be recommended to players before the UWR game.