RESUMO
Although it is appreciated that bacterial chemotaxis systems rely on coupling, also called scaffold, proteins to both connect input receptors with output kinases and build interkinase connections that allow signal amplification, it is not yet clear why many systems use more than one coupling protein. We examined the distinct functions for multiple coupling proteins in the bacterial chemotaxis system of Helicobacter pylori, which requires two nonredundant coupling proteins for chemotaxis: CheW and CheV1, a hybrid of a CheW and a phosphorylatable receiver domain. We report that CheV1 and CheW have largely redundant abilities to interact with chemoreceptors and the CheA kinase, and both similarly activated CheA's kinase activity. We discovered, however, that they are not redundant for formation of the higher order chemoreceptor arrays that are known to form via CheA-CheW interactions. In support of this possibility, we found that CheW and CheV1 interact with each other and with CheA independent of the chemoreceptors. Therefore, it seems that some microbes have modified array formation to require CheW and CheV1. Our data suggest that multiple coupling proteins may be used to provide flexibility in the chemoreceptor array formation.
Assuntos
Quimiotaxia/fisiologia , Transdução de Sinais , Sequência de Aminoácidos , Proteínas de Bactérias/metabolismo , Proteínas de Transporte/metabolismo , Proteínas Quimiotáticas Aceptoras de Metil/química , Proteínas Quimiotáticas Aceptoras de Metil/metabolismo , Fosforilação , Ligação ProteicaRESUMO
The epithelial layer of the gastrointestinal tract contains invaginations, called glands or crypts, which are colonized by symbiotic and pathogenic microorganisms and may function as designated niches for certain species. Factors that control gland colonization are poorly understood, but bacterial chemotaxis aids occupation of these sites. We report here that a Helicobacter pylori cytoplasmic chemoreceptor, TlpD, is required for gland colonization in the stomach. tlpD mutants demonstrate gland colonization defects characterized by a reduction in the percentage of glands colonized but not in the number of bacteria per gland. Consistent with TlpD's reported role in reactive oxygen species (ROS) avoidance, tlpD mutants showed hallmarks of exposure to high ROS. To assess the role of host-generated ROS in TlpD-dependent gland colonization, we utilized mice that lack either the ability to generate epithelial hydrogen peroxide or immune cell superoxide. tlpD gland colonization defects were rescued to wild-type H. pylori levels in both of these mutants. These results suggest that multiple types of innate immune-generated ROS production limit gland colonization and that bacteria have evolved specific mechanisms to sense and direct their motility in response to this signal and thus spread throughout tissue.
Assuntos
Quimiotaxia/fisiologia , Mucosa Gástrica/microbiologia , Helicobacter pylori/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Portador Sadio , Feminino , Regulação da Expressão Gênica , Genes Bacterianos , Humanos , Masculino , Camundongos , Camundongos Knockout , MutaçãoRESUMO
The current study examined the effect that game design modification, goal type, and player numbers on the running performance and physiological demands of small-sided hurling games (SSG). Forty-eight hurling players (age, 25.5 ± 3.2 years; height, 178.9 ± 3.2 cm; body mass, 78.5 ± 4.5 kg) performed 4 types of SSG (possession [P], normal play [NP], regular goals [RG] and small goals [SG]) in 4-a-side, 5-a-side, and 6-a-side formats. Heart rate (Polar Electro Oy) and global positioning system technology (VX Sport, 4-Hz, Lower Hutt) were used to analyze the physical and physiological differences between SSG. Total distance (m), high-speed running distance (m) (≥17 km·h), very-high speed running distance (≥22 km·h) (m), peak and mean velocity (km·h) were analyzed as an indicator of the physical demands of play. The 4-a-side SSG independent of game design and goal type resulted in a significantly higher relative exercise intensity compared with 5-a-side (mean change: 6 ± 2%; p = 0.001; d = 1.9 ± 0.2; large) and 6-a-side SSG independent of game design or goal type (mean change: 12 ± 2%; p = 0.001; d = 2.9 ± 0.8; very large). The 4-a-side SG (619 ± 106-m [419-735-m]) resulted in the highest distance when compared with all PP (mean change: 141 ± 9 m; p = 0.05; d = 1.9 ± 0.3; moderate) and RG (mean change: 119 ± 39 m; p = 0.004; d = 2.1 ± 0.8; large). Similar trends were observed for 5-a-side and 6-a-side games with SG resulting in increased total running performance. In conclusion, the current observations reveal that 4-a-side NP, SG, and RG have the highest physiological demands with 4-a-side SG having increased running performance in contrast to other game design and goal-type games. Furthermore, independent of game design and goal type, 4-a-side SSG show increased relative intensity compared with 5-a-side and 6-a-side SSG.
Assuntos
Desempenho Atlético/fisiologia , Esportes/fisiologia , Adulto , Atletas , Sistemas de Informação Geográfica , Frequência Cardíaca/fisiologia , Humanos , Masculino , Condicionamento Físico Humano , Aptidão Física/fisiologia , Corrida/fisiologiaRESUMO
The current study examined how the impact of pitch dimensions influences physiological and running performance during 4-minute small-sided games (SSGs). Twenty-four (n = 24) hurling players were monitored with global positioning system and heart rate monitors during the in-season training period. Total distance (in meters), high-speed running distance (in meters) (≥17 km·h), very high-speed running distance (≥22 km·h) (in meters), total accelerations (n), acceleration distance (in meters), and peak and mean velocity (in kilometers per hour) were calculated. Additionally, SSGs rate of perceived exertion (RPESSG; AU), % maximum heart rate, and individualized training impulse (iTRIMP; AU) were collected. The current results show that the manipulation of SSGs pitch size has an impact on the running performance and physiological responses. The data showed that SSGs played on large pitches (SSG80 × 20 m) had greater running demands than medium (SSG60 × 20 m) or small (SSG40 × 20 m) pitches, with significantly more distance covered in all movement categories. Total distance covered at high speed was 354 ± 111 m on a large pitch, 254 ± 72 m on a medium pitch, and 198 ± 62 m on a small pitch. Large pitch dimensions resulted in greater physiological and perceptual demands on players (higher %HRmax, iTRIMP [AU], and RPESSG [AU]) compared with medium and small pitches. The current data help applied practitioners to understand further how modifying different aspects of SSGs can alter the running and physiological responses of players. Moreover, applied practitioners now have consistent information to design and optimize their training time in mixing the physical, technical, and tactical elements within specific SSGs pitch dimensions.
Assuntos
Atletas , Desempenho Atlético/fisiologia , Esportes/fisiologia , Aceleração , Adulto , Sistemas de Informação Geográfica , Frequência Cardíaca/fisiologia , Humanos , Masculino , Movimento , Força Muscular , Esforço Físico/fisiologia , Corrida/fisiologia , Adulto JovemRESUMO
UNLABELLED: Cytoplasmic chemoreceptors are widespread among prokaryotes but are far less understood than transmembrane chemoreceptors, despite being implicated in many processes. One such cytoplasmic chemoreceptor is Helicobacter pylori TlpD, which is required for stomach colonization and drives a chemotaxis response to cellular energy levels. Neither the signals sensed by TlpD nor its molecular mechanisms of action are known. We report here that TlpD functions independently of the other chemoreceptors. When TlpD is the sole chemoreceptor, it is able to localize to the pole and recruits CheW, CheA, and at least two CheV proteins to this location. It loses the normal membrane association that appears to be driven by interactions with other chemoreceptors and with CheW, CheV1, and CheA. These results suggest that TlpD can form an autonomous signaling unit. We further determined that TlpD mediates a repellent chemotaxis response to conditions that promote oxidative stress, including being in the presence of iron, hydrogen peroxide, paraquat, and metronidazole. Last, we found that all tested H. pylori strains express TlpD, whereas other chemoreceptors were present to various degrees. Our data suggest a model in which TlpD coordinates a signaling complex that responds to oxidative stress and may allow H. pylori to avoid areas of the stomach with high concentrations of reactive oxygen species. IMPORTANCE: Helicobacter pylori senses its environment with proteins called chemoreceptors. Chemoreceptors integrate this sensory information to affect flagellum-based motility in a process called chemotaxis. Chemotaxis is employed during infection and presumably aids H. pylori in encountering and colonizing preferred niches. A cytoplasmic chemoreceptor named TlpD is particularly important in this process, and we report here that this chemoreceptor is able to operate independently of other chemoreceptors to organize a chemotaxis signaling complex and mediate a repellent response to oxidative stress conditions. H. pylori encounters and must cope with oxidative stress during infection due to oxygen and reactive oxygen species produced by host cells. TlpD's repellent response may allow the bacteria to escape niches experiencing inflammation and elevated reactive oxygen species (ROS) production.
Assuntos
Proteínas de Bactérias/metabolismo , Quimiotaxia/fisiologia , Regulação Bacteriana da Expressão Gênica/fisiologia , Helicobacter pylori/fisiologia , Proteínas de Bactérias/genética , Estresse Oxidativo , Transdução de SinaisRESUMO
Malone, S, Solan, B, Collins, KD, and Doran, DA. Positional match running performance in elite Gaelic football. J Strength Cond Res 30(8): 2292-2298, 2016-There is currently limited information available on match running performance in Gaelic football. The objective of the current study was to report on the match running profile of elite male Gaelic football and assess positional running performance. In this observational study, 50 elite male Gaelic football players wore 4-Hz global positioning systems units (VX Sports) across 30 competitive games with a total of 215 full game data sets collected. Activity was classed according to total distance, high-speed distance (≥17 km·h), sprint distance (≥22 km·h), mean velocity (km·h), peak velocity (km·h), and number of accelerations. The average match distance was 8,160 ± 1,482 m, reflective of a relative distance of 116 ± 21 m·min, with 1,731 ± 659 m covered at high speed, which is reflective of a relative high-speed distance of 25 ± 9 m·min. The observed sprint distance was 445 ± 169 m distributed across 44 sprint actions. The peak velocity was 30.3 ± 1.8 km·h with a mean velocity of 6.5 ± 1.2 km·h. Players completed 184 ± 40 accelerations, which represent 2.6 ± 0.5 accelerations per minute. There were significant differences between positional groups for both total running distance, high-speed running distance, and sprint distance, with midfielders covering more total and high-speed running distance, compared with other positions (p < 0.001). There was a reduction in high-speed and sprint distance between the first and second half (p < 0.001). Reductions in running performance were position dependent with the middle 3 positions experiencing the highest decrement in performance. The current study is the first to communicate a detailed description of match running performance during competitive elite Gaelic football match play.
Assuntos
Desempenho Atlético/fisiologia , Futebol Americano/fisiologia , Sistemas de Informação Geográfica , Corrida/fisiologia , Aceleração , Humanos , Masculino , Adulto JovemRESUMO
PURPOSE: To examine the association between combined session rating of perceived exertion (RPE) workload measures and injury risk in elite Gaelic footballers. METHODS: Thirty-seven elite Gaelic footballers (mean ± SD age 24.2 ± 2.9 y) from 1 elite squad were involved in a single-season study. Weekly workload (session RPE multiplied by duration) and all time-loss injuries (including subsequent-wk injuries) were recorded during the period. Rolling weekly sums and wk-to-wk changes in workload were measured, enabling the calculation of the acute:chronic workload ratio by dividing acute workload (ie, 1-weekly workload) by chronic workload (ie, rolling-average 4-weekly workload). Workload measures were then modeled against data for all injuries sustained using a logistic-regression model. Odds ratios (ORs) were reported against a reference group. RESULTS: High 1-weekly workloads (≥2770 arbitrary units [AU], OR = 1.63-6.75) were associated with significantly higher risk of injury than in a low-training-load reference group (<1250 AU). When exposed to spikes in workload (acute:chronic workload ratio >1.5), players with 1 y experience had a higher risk of injury (OR = 2.22) and players with 2-3 (OR = 0.20) and 4-6 y (OR = 0.24) of experience had a lower risk of injury. Players with poorer aerobic fitness (estimated from a 1-km time trial) had a higher injury risk than those with higher aerobic fitness (OR = 1.50-2.50). An acute:chronic workload ratio of (≥2.0) demonstrated the greatest risk of injury. CONCLUSIONS: These findings highlight an increased risk of injury for elite Gaelic football players with high (>2.0) acute:chronic workload ratios and high weekly workloads. A high aerobic capacity and playing experience appears to offer injury protection against rapid changes in workload and high acute:chronic workload ratios. Moderate workloads, coupled with moderate to high changes in the acute:chronic workload ratio, appear to be protective for Gaelic football players.
Assuntos
Condicionamento Físico Humano , Esforço Físico/fisiologia , Aptidão Física/fisiologia , Futebol/lesões , Comportamento Competitivo/fisiologia , Humanos , Masculino , Percepção , Estudos Prospectivos , Fatores de Risco , Estações do Ano , Adulto JovemRESUMO
OBJECTIVES: To examine the association between combined sRPE measures and injury risk in elite professional soccer. DESIGN: Observational cohort study. METHODS: Forty-eight professional soccer players (mean±SD age of 25.3±3.1 yr) from two elite European teams were involved within a one season study. Players completed a test of intermittent-aerobic capacity (Yo-YoIR1) to assess player's injury risk in relation to intermittent aerobic capacity. Weekly workload measures and time loss injuries were recorded during the entire period. Rolling weekly sums and week-to-week changes in workload were measured, allowing for the calculation of the acute:chronic workload ratio, which was calculated by dividing the acute (1-weekly) and chronic (4-weekly) workloads. All derived workload measures were modelled against injury data using logistic regression. Odds ratios (OR) were reported against a reference group. RESULTS: Players who exerted pre-season 1-weekly loads of ≥1500 to ≤2120AU were at significantly higher risk of injury compared to the reference group of ≤1500AU (OR=1.95, p=0.006). Players with increased intermittent-aerobic capacity were better able to tolerate increased 1-weekly absolute changes in training load than players with lower fitness levels (OR=4.52, p=0.011). Players who exerted in-season acute:chronic workload ratios of >1.00 to <1.25 (OR=0.68, p=0.006) were at significantly lower risk of injury compared to the reference group (≤0.85). CONCLUSIONS: These findings demonstrate that an acute:chronic workload of between 1.00 and 1.25 is protective for professional soccer players. A higher intermittent-aerobic capacity appears to offer greater injury protection when players are exposed to rapid changes in workload in elite soccer players. Moderate workloads, coupled with moderate-low to moderate-high acute:chronic workload ratios, appear to be protective for professional soccer players.
Assuntos
Traumatismos em Atletas/etiologia , Esforço Físico , Futebol/lesões , Carga de Trabalho , Adulto , Traumatismos em Atletas/prevenção & controle , Humanos , Modelos Logísticos , Razão de Chances , Estudos Prospectivos , Fatores de Proteção , Fatores de Risco , Futebol/fisiologia , Fatores de TempoRESUMO
SUMMARY: Chemoreceptors sense environmental signals and drive chemotactic responses in Bacteria and Archaea. There are two main classes of chemoreceptors: integral inner membrane and soluble cytoplasmic proteins. The latter were identified more recently than integral membrane chemoreceptors and have been studied much less thoroughly. These cytoplasmic chemoreceptors are the subject of this review. Our analysis determined that 14% of bacterial and 43% of archaeal chemoreceptors are cytoplasmic, based on currently sequenced genomes. Cytoplasmic chemoreceptors appear to share the same key structural features as integral membrane chemoreceptors, including the formations of homodimers, trimers of dimers, and 12-nm hexagonal arrays within the cell. Cytoplasmic chemoreceptors exhibit varied subcellular locations, with some localizing to the poles and others appearing both cytoplasmic and polar. Some cytoplasmic chemoreceptors adopt more exotic locations, including the formations of exclusively internal clusters or moving dynamic clusters that coalesce at points of contact with other cells. Cytoplasmic chemoreceptors presumably sense signals within the cytoplasm and bear diverse signal input domains that are mostly N terminal to the domain that defines chemoreceptors, the so-called MA domain. Similar to the case for transmembrane receptors, our analysis suggests that the most common signal input domain is the PAS (Per-Arnt-Sim) domain, but a variety of other N-terminal domains exist. It is also common, however, for cytoplasmic chemoreceptors to have C-terminal domains that may function for signal input. The most common of these is the recently identified chemoreceptor zinc binding (CZB) domain, found in 8% of all cytoplasmic chemoreceptors. The widespread nature and diverse signal input domains suggest that these chemoreceptors can monitor a variety of cytoplasmically based signals, most of which remain to be determined.