Effects of situational variables on the physical activity profiles of elite soccer players in different score line states.

The aim of this study were to investigate the effects of playing position, pitch location, team ability and opposition ability on the physical activity profiles of English premier league soccer players in difference score line states. A validated automatic tracking system (Venatrack Ltd.) was used to track players in real time (at 25 Hz) for total distance covered, high speed running distance and sprint distance. This is the first study to include every team from an entire season in the English premier league, resulting in 376 games, 570 players and 35 000 rows of data from the 2011-12 season being analyzed using multi-level modelling. Multi-level regression revealed an inverted "u" shaped association between total distance covered and goal difference (GD), with greater distances covered when GD was zero and reduced distances when GD was either positive or negative. A similar "u" shaped association was found with high speed distance covered at home. In addition distance covered (both at home and away) were predicted by playing position. All activity profiles (with the exception of sprint distance at home) were predicted by pitch location and time scored. Lastly, distance away from home and high speed running at home were predicted by opposition ability. Score line appears to effect player activity profiles across a number of situational factors and thus should be considered by managers when preparing and selecting teams to maximize performance. The current study also highlighted the need for more sensitive score line definitions in which to consider score line effects.

Influence of lung volume on the interaction between cardiac output and cerebrovascular regulation during extreme apnoea.

NEW FINDINGS: What is the central question of this study? Does the reduction in cardiac output observed during extreme voluntary apnoea, secondary to high lung volume, result in a reduction in cerebral blood flow, perfusion pressure and oxygen delivery in a group of elite free divers? What is the main finding and its importance? High lung volumes reduce cardiac output and ventricular filling during extreme apnoea, but changes in cerebral blood flow are observed only transiently during the early stages of apnoea. This reveals that whilst cardiac output is important in regulating cerebral haemodynamics, the role of mean arterial pressure in restoring cerebral perfusion pressure is of greater significance to the regulation of cerebral blood flow. We investigated the role of lung volume-induced changes in cardiac output (Q̇) on cerebrovascular regulation during prolonged apnoea. Fifteen elite apnoea divers (one female; 185 ± 7 cm, 82 ± 12 kg, 29 ± 7 years old) attended the laboratory on two separate occasions and completed maximal breath-holds at total lung capacity (TLC) and functional residual capacity (FRC) to elicit disparate cardiovascular responses. Mean arterial pressure (MAP), internal jugular venous pressure and arterial blood gases were measured via cannulation. Global cerebral blood flow was quantified by ultrasound and cardiac output was quantified by via photoplethysmography. At FRC, stroke volume and Q̇ did not change from baseline (P > 0.05). In contrast, during the TLC trial stroke volume and Q̇ were decreased until 80 and 40% of apnoea, respectively (P < 0.05). During the TLC trial, global cerebral blood flow was significantly lower at 20%, but subsequently increased so that cerebral oxygen delivery was comparable to that during the FRC trial. Internal jugular venous pressure was significantly higher throughout the TLC trial in comparison to FRC. The MAP increased progressively in both trials but to a greater extent at TLC, resulting in a comparable cerebral perfusion pressure between trials by the end of apnoea. In summary, although lung volume has a profound effect on Q̇ during prolonged breath-holding, these changes do not translate to the cerebrovasculature owing to the greater sensitivity of cerebral blood flow to arterial blood gases and MAP; regulatory mechanisms that facilitate the maintenance of cerebral oxygen delivery.

Hydrogenosomes of laboratory-induced metronidazole-resistant Trichomonas vaginalis lines are downsized while those from clinically metronidazole-resistant isolates are not.

Trichomonas vaginalis is the most common sexually transmitted protozoan in the world and its resistance to metronidazole is increasing. The purpose of this study was to demonstrate that clinical metronidazole resistance in T. vaginalis does not occur via the same mechanism as laboratory-induced metronidazole resistance--that is, via hydrogenosome down sizing. Ultrathin sections of this parasite were examined using transmission electron microscopy and the size and area of the cell and hydrogenosomes were compared between drug-resistant laboratory lines and clinically resistant isolates. Clinical metronidazole-resistant T. vaginalis had similar-sized hydrogenosomes as a metronidazole-sensitive isolate. Inducing metronidazole resistance in both of these isolates caused down sizing of hydrogenosomes. Inducing toyocamycin resistance did not cause any ultrastructural changes to the cell or to the hydrogenosome. No correlation between hydrogenosome number and the drug-resistant status of T. vaginalis isolates and lines was observed. This report demonstrates that clinical metronidazole resistance is not associated with down-sized hydrogenosomes, thus indicating that an alternative resistance mechanism is used by T. vaginalis.

Investigation of the regulation of transcriptional changes in Ancylostoma caninum larvae following serum activation, with a focus on the insulin-like signalling pathway.

The exit from dauer in the free-living nematode Caenorhabditis elegans is under the control of a single amphidial neuron (ASJ) of the insulin-like signalling pathway. Mutations of this pathway have the ability to suppress entry into the dauer stage. It has been postulated that insulin-like signalling plays a significant role in the response to serum stimulation in vitro of the third-stage larvae (L3s) of the canine hookworm Ancylostoma caninum. To test for the possible involvement of the insulin-like signalling cascade in the response to serum stimulation, the effects of two signalling stimulants (8-bromo cGMP and arecoline) and four inhibitors, namely 4,7-phenanthroline, phosphoinositide-3 kinase (PI3K), Akt inhibitor IV and rapamycin on feeding and on levels of selected activation-associated mRNAs in serum-stimulated L3s were explored. L3s of A. caninum were pre-incubated with or without the appropriate inhibitor/agonist. Following serum-stimulation, the feeding activity was assessed. The transcription levels of a number of activation-associated mRNAs linked to particular expressed sequence tags (ESTs) were investigated by reverse transcription, real-time PCR (rtPCR). The treatment of worms with 4,7-phenanthroline completely suppressed feeding and significantly reduced the differential levels of most activation-associated mRNAs, whereas the treatment with cGMP resulted in the resumption of feeding in almost 85% of the L3s and yielded a specific transcriptional profile consistent with that following serum stimulation. The treatment of L3s with arecoline resulted in the resumption of feeding in approximately 85% of L3s, but did not result in a transcriptomic profile consistent with activation. A complete reduction in feeding was recorded in the presence of the PI3K inhibitor LY294002 (1mM) and resulted in a pronounced dampening of differential transcription in response to serum stimulation for the molecules examined. Akt inhibitor IV resulted in a approximately 70% reduction in feeding but had almost no effect on the level of any of the activation-associated mRNAs studied. Rapamycin was shown to have a weak effect on feeding, and several of the mRNAs studied exhibited greater than expected transcription following treatment. The complexities of activation-associated transcription could not be addressed using the current approach. A larger number of mRNAs needs to be investigated in order to predict or identify regulatory mechanisms proposed to function in the insulin-like signalling pathway in A. caninum.

Effects of playing position, pitch location, opposition ability and team ability on the technical performance of elite soccer players in different score line states.

The purpose of this study was to investigate the effects of playing position, pitch location, team ability and opposition ability on technical performance variables (pass, cross, corner, free kick accuracy) of English Premier League Soccer players in difference score line states. A validated automatic tracking system (Venatrack) was used to code player actions in real time for passing accuracy, cross accuracy, corner accuracy and free kick accuracy. In total 376 of the 380 games played during the 2011-12 English premier League season were recorded, resulting in activity profiles of 570 players and over 35'000 rows of data. These data were analysed using multi-level modelling. Multi-level regression revealed a "u" shaped association between passing accuracy and goal difference (GD) with greater accuracy occurring at extremes of GD e.g., when the score was either positive or negative. The same pattern was seen for corner accuracy away from home e.g., corner accuracy was lowest when the score was close with the lowest accuracy at extremes of GD. Although free kicks were not associated with GD, team ability, playing position and pitch location were found to predict accuracy. No temporal variables were found to predict cross accuracy. A number of score line effects were present across the temporal factors which should be considered by coaches and managers when preparing and selecting teams in order to maximise performance. The current study highlighted the need for more sensitive score line definitions in which to consider score line effects.

Time-motion analysis of first-class cricket fielding.

This study aimed to quantify player movements during first-class cricket fielding. Using real-time computerised time-motion analysis the entire on-field activities of 27 in-fielders were observed for 10-over periods; 9 during each of the morning, afternoon and evening sessions of first-class cricket. In addition 8 first-class cricketers performed 15m speed trials between timing gates to provide velocity multipliers for distance estimation. Overall, players changed movement every 6.4+/-1.1s (mean+/-S.D.) and fielded the ball 0.5+/-0.4 times per over. Stationary and walking activity represented 94.2+/-2.4% of match time. High-intensity (HI) activity represented 1.6+/-0.8% of match time with mean burst and recovery durations of 1.3+/-0.3 and 99.8+/-94.5s, respectively. Repeated HI bouts (at least 3 bursts with less than 21s mean recovery) occurred 1.2 times per 10-over period. Fielders covered an estimated 15.5km per day. In conclusion, first-class fielding entails less HI activity than other team sports such as soccer and hockey. However, fielders are required to cover large distances in a day, but over 77% of these distances are covered by walking.

Effective speed and agility conditioning methodology for random intermittent dynamic type sports.

Different coaching methods are often used to improve performance. This study compared the effectiveness of 2 methodologies for speed and agility conditioning for random, intermittent, and dynamic activity sports (e.g., soccer, tennis, hockey, basketball, rugby, and netball) and the necessity for specialized coaching equipment. Two groups were delivered either a programmed method (PC) or a random method (RC) of conditioning with a third group receiving no conditioning (NC). PC participants used the speed, agility, quickness (SAQ) conditioning method, and RC participants played supervised small-sided soccer games. PC was also subdivided into 2 groups where participants either used specialized SAQ equipment or no equipment. A total of 46 (25 males and 21 females) untrained participants received (mean +/- SD) 12.2 +/- 2.1 hours of physical conditioning over 6 weeks between a battery of speed and agility parameter field tests. Two-way analysis of variance results indicated that both conditioning groups showed a significant decrease in body mass and body mass index, although PC achieved significantly greater improvements on acceleration, deceleration, leg power, dynamic balance, and the overall summation of % increases when compared to RC and NC (p < 0.05). PC in the form of SAQ exercises appears to be a superior method for improving speed and agility parameters; however, this study found that specialized SAQ equipment was not a requirement to observe significant improvements. Further research is required to establish whether these benefits transfer to sport-specific tasks as well as to the underlying mechanisms resulting in improved performance.

Physical Demands of Different Positions in FA Premier League Soccer.

The purpose of this study was to evaluate the physical demands of English Football Association (FA) Premier League soccer of three different positional classifications (defender, midfielder and striker). Computerised time-motion video-analysis using the Bloomfield Movement Classification was undertaken on the purposeful movement (PM) performed by 55 players. Recognition of PM had a good inter-tester reliability strength of agreement (κ= 0.7277). Players spent 40.6 ± 10.0% of the match performing PM. Position had a significant influence on %PM time spent sprinting, running, shuffling, skipping and standing still (p < 0.05). However, position had no significant influence on the %PM time spent performing movement at low, medium, high or very high intensities (p > 0.05). Players spent 48.7 ± 9.2% of PM time moving in a directly forward direction, 20.6 ± 6.8% not moving in any direction and the remainder of PM time moving backward, lateral, diagonal and arced directions. The players performed the equivalent of 726 ± 203 turns during the match; 609 ± 193 of these being of 0° to 90° to the left or right. Players were involved in the equivalent of 111 ± 77 on the ball movement activities per match with no significant differences between the positions for total involvement in on the ball activity (p > 0.05). This study has provided an indication of the different physical demands of different playing positions in FA Premier League match-play through assessment of movements performed by players. Key pointsPlayers spent ~40% of the match performing Pur-poseful Movement (PM).Position had a significant influence on %PM time spent performing each motion class except walking and jogging. Players performed >700 turns in PM, most of these being of 0°-90°.Strikers performed most high to very high intensity activity and most contact situations.Defenders also spent a significantly greater %PM time moving backwards than the other two posi-tions.Different positions could benefit from more specific conditioning programs.

Haemoprotozoa: Making biological sense of molecular phylogenies.

A range of protistan parasites occur in the blood of vertebrates and are transmitted by haematophagous invertebrate vectors. Some 48 genera are recognized in bood primarily on the basis of parasite morphology and host specificity; including extracellular kinetoplastids (trypanosomatids) and intracellular apicomplexa (haemogregarines, haemococcidia, haemosporidia and piroplasms). Gene sequences are available for a growing number of species and molecular phylogenies often link parasite and host or vector evolution. This review endeavours to reconcile molecular clades with biological characters. Four major trypanosomatid clades have been associated with site of development in the vector: salivarian or stercorarian for Trypanosoma, and supra- or peri-pylorian for Leishmania. Four haemogregarine clades have been associated with acarine vectors (Hepatozoon A and B, Karyolysus, Hemolivia) and another two with leeches (Dactylosoma, Haemogregarina sensu stricto). Two haemococcidian clades (Lankesterella, Schellackia) using leeches and mosquitoes (as paratenic hosts!) were paraphyletic with monoxenous enteric coccidia. Two major haemosporidian clades have been associated with mosquito vectors (Plasmodium from mammals, Plasmodium from birds and lizards), two with midges (Hepatocystis from bats, Parahaemoproteus from birds) and two with louse-flies and black-flies (Haemoproteus and Leucocytozoon from birds). Three major piroplasm clades were recognized: one associated with transovarian transmission in ticks (Babesia sensu stricto); one with pre-erythrocytic schizogony in vertebrates (Theileria/Cytauxzoon); and one with neither (Babesia sensu lato). Broad comparative studies with allied groups suggest that trypanosomatids and haemogregarines evolved first in aquatic and then terrestrial environments, as evidenced by extant lineages in invertebrates and their radiation in vertebrates. In contrast, haemosporidia and haemococcidia are thought to have evolved first in vertebrates from proto-coccidia and then incorporated invertebrate vectors. Piroplasms are thought to have evolved in ticks and diversified into mammals. More molecular studies are required on more parasite taxa to refine current thought, but ultimately transmission studies are mandated to determine the vectors for many haemoprotozoa.

Ultrastructural and Molecular Characterisation of an Heterosporis-Like Microsporidian in Australian Sea Snakes (Hydrophiinae).

Four sea snakes (two Hydrophis major, one Hydrophis platurus, one Hydrophis elegans) were found washed ashore on different beaches in the Sunshine Coast region and Fraser Island in Queensland, Australia between 2007-2013. Each snake had multiple granulomas and locally extensive regions of pallor evident in the hypaxial and intercostal musculature along the body. Lesions in two individuals were also associated with vertebral and rib fractures. Histological examination revealed granulomas scattered throughout skeletal muscle, subcutaneous adipose tissue and fractured bone. These were composed of dense aggregates of microsporidian spores surrounded by a mantle of macrophages. Sequences (ssrRNA) were obtained from lesions in three sea snakes and all revealed 99% similarity with Heterosporis anguillarum from the Japanese eel (Anguillarum japonica). However, ultrastructural characteristics of the organism were not consistent with those of previous descriptions. Electron microscopic examination of skeletal muscle revealed large cysts (not xenomas) bound by walls of fibrillar material (Heterosporis-like sporophorocyst walls were not detected). The cysts contained numerous mature microsporidian spores arranged in small clusters, sometimes apparently within sporophorous vesicles. The microspores were monomorphic, oval and measured 2.5-3.0 µm by 1.6-1.8 µm. They contained isofilar polar filaments with 11 (infrequently 9-12) coils arranged in two ranks. This is the first published report of a microsporidian infection in hydrophiid sea snakes. This discovery shows microsporidia with molecular affinities to Heterosporis anguillarum but ultrastructural characters most consistent with the genus Pleistophora (but no hitherto described species). Further studies are required to determine whether the microsporidian presented here belongs to the genus Heterosporis, or to a polymorphic species group as suggested by the recognition of a robust Pleistophora/Heterosporis clade by molecular studies. The gross and histological pathology associated with these infections are described.

A retrospective study of Babesia macropus associated with morbidity and mortality in eastern grey kangaroos (Macropus giganteus) and agile wallabies (Macropus agilis).

This is a retrospective study of 38 cases of infection by Babesia macropus, associated with a syndrome of anaemia and debility in hand-reared or free-ranging juvenile eastern grey kangaroos (Macropus giganteus) from coastal New South Wales and south-eastern Queensland between 1995 and 2013. Infection with B. macropus is recorded for the first time in agile wallabies (Macropus agilis) from far north Queensland. Animals in which B. macropus infection was considered to be the primary cause of morbidity had marked anaemia, lethargy and neurological signs, and often died. In these cases, parasitised erythrocytes were few or undetectable in peripheral blood samples but were sequestered in large numbers within small vessels of visceral organs, particularly in the kidney and brain, associated with distinctive clusters of extraerythrocytic organisms. Initial identification of this piroplasm in peripheral blood smears and in tissue impression smears and histological sections was confirmed using transmission electron microscopy and molecular analysis. Samples of kidney, brain or blood were tested using PCR and DNA sequencing of the 18S ribosomal RNA and heat shock protein 70 gene using primers specific for piroplasms. The piroplasm detected in these samples had 100% sequence identity in the 18S rRNA region with the recently described Babesia macropus in two eastern grey kangaroos from New South Wales and Queensland, and a high degree of similarity to an unnamed Babesia sp. recently detected in three woylies (Bettongia penicillata ogilbyi) in Western Australia.

Drug resistance in the sexually transmitted protozoan Trichomonas vaginalis.

Trichomoniasis is the most common, sexually transmitted infection. It is caused by the flagellated protozoan parasite Trichomonas vaginalis. Symptoms include vaginitis and infections have been associated with preterm delivery, low birth weight and increased infant mortality, as well as predisposing to HIV/AIDS and cervical cancer. Trichomoniasis has the highest prevalence and incidence of any sexually transmitted infection. The 5-nitroimidazole drugs, of which metronidazole is the most prescribed, are the only approved, effective drugs to treat trichomoniasis. Resistance against metronidazole is frequently reported and cross-resistance among the family of 5-nitroimidazole drugs is common, leaving no alternative for treatment, with some cases remaining unresolved. The mechanism of metronidazole resistance in T. vaginalis from treatment failures is not well understood, unlike resistance which is developed in the laboratory under increasing metronidazole pressure. In the latter situation, hydrogenosomal function which is involved in activation of the prodrug, metronidazole, is down-regulated. Reversion to sensitivity is incomplete after removal of drug pressure in the highly resistant parasites while clinically resistant strains, so far analysed, maintain their resistance levels in the absence of drug pressure. Although anaerobic resistance has been regarded as a laboratory induced phenomenon, it clearly has been demonstrated in clinical isolates. Pursuit of both approaches will allow dissection of the underlying mechanisms. Many alternative drugs and treatments have been tested in vivo in cases of refractory trichomoniasis, as well as in vitro with some successes including the broad spectrum anti-parasitic drug nitazoxanide. Drug resistance incidence in T. vaginalis appears to be on the increase and improved surveillance of treatment failures is urged.

Phylogeny and biogeography of the "Australian" trichostomes (Ciliophora: Litostomata).

The phylogenetic relationships of members of the ciliate class Litostomatea were determined by a molecular phylogeny using the small subunit of the ribosomal RNA (ssu-rRNA) gene and a morphological phylogeny based on ultrastructural analyses of the group. Molecular analyses consistently supported the monophyly of Trichostomatia, Entodiniomorphida and the "Australian" trichostomes but provided limited support for a monophyletic Vestibuliferida and Haptoria. The results of the morphological analyses depended on the way in which the dataset was treated: "unordered" and "ordered" recovered a monophyletic Trichostomatia, Haptoria and the "Australian" trichostomes but challenged the monophyly of Entodinimorphida and Vestibuliferida; "dollo" recovered a monophyletic Trichostomatia and Entodiniomorphida but at the cost of a greatly longer tree than either "unordered" or "ordered" datasets. The monophyly of each "Australian" trichostome family was supported in all analyses and by both approaches. These results suggest that the trichostome ciliates may have become associated with mammals in Gondwana with the "Australian" trichostome ciliates entering Australia with primitive herbivorous marsupials. Subsequent diversification of the "Australian" families was probably a result of dietary specialization and oral and cortical synapomorphies define each family. We decline at this time to erect a formal taxon name for the "Australian" trichostomes due to the instability of other superfamilial taxa within the Litosomatea and concerns about the stability of tree topology until a better taxon sample of litostome ciliates is available.

A pore-forming haemolysin from the hookworm, Ancylostoma caninum.

Hookworms feed on blood, but the mechanism by which they lyse ingested erythrocytes is unknown. Here we show that Ancylostoma caninum, the common dog hookworm, expresses a detergent soluble, haemolytic factor. Activity was identified in both adult and larval stages, was heat-stable and unaffected by the addition of protease inhibitors, metal ions, chelators and reducing agents. Trypsin ablated lysis indicating that the haemolysin is a protein. A closely migrating doublet of hookworm proteins with apparent molecular weights of 60-65 kDa bound to the erythrocyte membrane after lysis of cells using both unlabeled and biotinylated detergent-solubilised hookworm extracts. In addition, separation of detergent-soluble parasite extracts using strong cation-exchange chromatography, resulted in purification of 60-65 kDa proteins with trypsin-sensitive haemolytic activity. Erythrocytes lysed with particulate, buffer-insoluble worm extracts were observed using scanning electron microscopy and appeared as red cell ghosts with approximately 100 nm diameter pores formed in the cell membranes. Red blood cell ghosts remained visible indicating that lysis was likely caused by pore formation and followed by osmotic disruption of the cell.

Transcriptional changes in the hookworm, Ancylostoma caninum, during the transition from a free-living to a parasitic larva.

BACKGROUND: Third-stage larvae (L3) of the canine hookworm, Ancylostoma caninum, undergo arrested development preceding transmission to a host. Many of the mRNAs up-regulated at this stage are likely to encode proteins that facilitate the transition from a free-living to a parasitic larva. The initial phase of mammalian host invasion by A. caninum L3 (herein termed "activation") can be mimicked in vitro by culturing L3 in serum-containing medium. METHODOLOGY/PRINCIPAL FINDINGS: The mRNAs differentially transcribed between activated and non-activated L3 were identified by suppression subtractive hybridisation (SSH). The analysis of these mRNAs on a custom oligonucleotide microarray printed with the SSH expressed sequence tags (ESTs) and publicly available A. caninum ESTs (non-subtracted) yielded 602 differentially expressed mRNAs, of which the most highly represented sequences encoded members of the pathogenesis-related protein (PRP) superfamily and proteases. Comparison of these A. caninum mRNAs with those of Caenorhabditis elegans larvae exiting from developmental (dauer) arrest demonstrated unexpectedly large differences in gene ontology profiles. C. elegans dauer exiting L3 up-regulated expression of mostly intracellular molecules involved in growth and development. Such mRNAs are virtually absent from activated hookworm larvae, and instead are over-represented by mRNAs encoding extracellular proteins with putative roles in host-parasite interactions. CONCLUSIONS/SIGNIFICANCE: Although this should not invalidate C. elegans dauer exit as a model for hookworm activation, it highlights the limitations of this free-living nematode as a model organism for the transition of nematode larvae from a free-living to a parasitic state.

Season of birth distribution of elite tennis players.

In many competitive sports, the use of a cut-off date for junior competition has been associated with a skewed birth date distribution in junior and senior players. The International Tennis Federation uses a junior competition year that commences on 1 January. The purpose of the current investigation was to describe the birth date distribution of 448 elite senior tennis players and 476 elite junior tennis players. There was a significant season of birth bias among elite senior players (P < 0.001), with 58.9% being born in the first 6 months of the year. There was also a significant season of birth bias among elite junior players (P < 0.001), with 59.5% being born in the first 6 months of the year. This pattern was observed in both male and female players as well as in players from different regions. The results provide evidence that it is the cut-off date for the junior competition year that is responsible for the skewed birth date distribution in tennis rather than regional or climatic factors.