Movement Demands of Elite Under-20s and Senior International Rugby Union Players

This study compared the movement demands of elite international Under-20 age grade (U20s) and senior international rugby union players during competitive tournament match play. Forty elite professional players from an U20 and 27 elite professional senior players from international performance squads were monitored using 10Hz global positioning systems (GPS) during 15 (U20s) and 8 (senior) international tournament matches during the 2014 and 2015 seasons. Data on distances, velocities, accelerations, decelerations, high metabolic load (HML) distance and efforts, and number of sprints were derived. Data files from players who played over 60 min (n = 258) were separated firstly into Forwards and Backs, and more specifically into six positional groups; FR–Front Row (prop & hooker), SR–Second Row, BR–Back Row (Flankers & No.8), HB–Half Backs (scrum half & outside half), MF–Midfield (centres), B3 –Back Three (wings & full back) for match analysis. Linear mixed models revealed significant differences between U20 and senior teams in both the forwards and backs. In the forwards the seniors covered greater HML distance (736.4 ± 280.3 vs 701.3 ± 198.7m, p = 0.01) and severe decelerations (2.38 ± 2.2 vs 2.28 ± 1.65, p = 0.05) compared to the U20s, but performed less relative HSR (3.1 ± 1.6 vs 3.2 ± 1.5, p < 0.01), moderate (19.4 ± 10.5 vs 23.6 ± 10.5, p = 0.01) and high accelerations (2.2 ± 1.9 vs 4.3 ± 2.7, p < 0.01) and sprint•min^-1 (0.11 ± 0.06 vs 0.11 ± 0.05, p < 0.01). Senior backs covered a greater relative distance (73.3 ± 8.1 vs 69.1 ± 7.6 m•min^-1, p < 0.01), greater High Metabolic Load (HML) distance (1138.0 ± 233.5 vs 1060.4 ± 218.1m, p < 0.01), HML efforts (112.7 ± 22.2 vs 98.8 ± 21.7, p < 0.01) and heavy decelerations (9.9 ± 4.3 vs 9.5 ± 4.4, p = 0.04) than the U20s backs. However, the U20s backs performed more relative HSR (7.3 ± 2.1 vs 7.2 ± 2.1, p <0.01) and sprint•min^-1 (0.26 ± 0.07 vs 0.25 ± 0.07, p < 0.01). Further investigation highlighted differences between the 6 positional groups of the teams. The positional groups that differed the most on the variables measured were the FR and MF groups, with the U20s FR having higher outputs on HSR, moderate & high accelerations, moderate, high & severe decelerations, HML distance, HML efforts, and sprints•min^-1. For the MF group the senior players produced greater values for relative distance covered, HSR, moderate decelerations, HML distance and sprint•min^-1. The BR position group was most similar with the only differences seen on heavy accelerations (U20s higher) and moderate decelerations (seniors higher). Findings demonstrate that U20s internationals appear to be an adequate ‘stepping stone’ for preparing players for movement characteristics found senior International rugby, however, the current study highlight for the first time that certain positional groups may require more time to be able to match the movement demands required at a higher playing level than others. Conditioning staff must also bear in mind that the U20s players whilst maintaining or improving match movement capabilities may require to gain substantial mass in some positions to match their senior counterparts.     

Human in vitro allogeneic responses. Demonstration of three pathways of T helper cell activation

In our study, we have measured in vitro proliferation and IL-2 production by human PBL to characterize the interactions between Th cells and accessory cells (AC) involved in responses to either conventional Ag or alloantigens. IL-2 production and proliferative responses to conventional Ag, such as influenza or tetanus, are exclusively dependent on the presence of CD4+ T cells and AC. In contrast, IL-2 and proliferative responses to alloantigen can be mediated by either CD4+ or CD8+ T cells. CD4+ T cells respond to alloantigen using either autologous AC (self-restricted), or allogeneic AC (allo-restricted), whereas CD8+ T cells respond to alloantigen using allogeneic AC only. The understanding of Th cell-AC interactions involved in in vitro allogeneic responses will be important for delineating the Th cell-AC interactions involved in transplantation immunity as well as in clinical disorders characterized by T cell dysfunction such as human immunodeficiency virus infection and systemic lupus erythematosus.     

Interleukin 2 receptor dysfunction in mice undergoing a graft-vs-host reaction

Mice of the (C57BL/10 X B10.A)F1 combination were given a single i.v. inoculation of 3 to 4 X 10(7) B10.A spleen cells, which induces a graft-vs-host (GVH)-associated immune deficiency in F1 mice. Between 1 and 4 wk later, spleen cells from the F1 mice were tested for the expression of IL 2 receptors by flow microfluorometry, using the 7D4 rat monoclonal antibody directed against an epitope murine IL 2 receptor. A reduction in intensity of spleen cell staining with 7D4 was detected as early as 8 days after parental cell inoculation, and no IL 2 receptors were detected by 28 days after initiation of GVH. Furthermore, the loss of IL 2 receptors was correlated with abrogation of proliferative responses to concanavalin A and lipopolysaccharide, of IL 2 production, and of cytotoxic T lymphocyte responses. These observations may be relevant for our understanding of GVH reactions, of immune disorders associated with GVH, and possibly of primary and acquired immunodeficiencies in general.     

Experimental allergic orchitis in mice

Inbred strains of mice were studied for their susceptibility to the induction of experimental allergic orchitis after sensitization with mouse testicular homogenate in complete Freund's adjuvant accompanied by injections of extract from Bordetella pertussis. Susceptibility to autoimmune orchitis was found to be linked to the major histocompatibility complex in BALB/c and C57BL/10 mice and mapped to genes encoded within the H-2D ^dregion. In five of six groups of bidirectional (susceptible × resistant) F₁ hybrids, H-2D ^d-linked susceptibility was inherited as a dominant autosomal trait. However, in (BALB/cByJ × DBA/2J)F₁ and (DBA/2J × BALB/cByJ)F₁ hybrids, dominant autosomal resistance to the induction of autoimmune orchitis was observed. Backcross analysis between the resistant F₁ hybrid and the susceptible BALB/cByJ parent suggests that a single independently segregating DBA/2J locus is capable of negating H-2D ^d-linked susceptibility, and controls resistance to the induction of autoimmune orchitis.Abbreviations used in this paper BP extract Bordetella pertussis extract - CFA complete Freund's adjuvant - EAO experimental allergic orchitis - Ir immune response - MHC major histocompatibility complex - MLH mouse liver homogenate - MTH mouse testis homogenate - PI pathology index     

Phorbol ester plus calcium ionophore induces release of arachidonic acid from membrane phospholipids of a human B cell line

Binding of LA350, a lymphoblastoid human B cell line, by phorbol myristate acetate (PMA) plus a calcium ionophore, either ionomycin or A23187, produced unique alterations in the release of arachidonic acid (AA) from cellular phospholipids. After equilibrium labeling of cells with radioactive fatty acids, [14C]AA demonstrated a selective enhanced release from the cells in response to the binding of PMA plus calcium ionophore as compared to the release of [14C]stearic acid (STE), [3H]oleic acid (OLE) and [3H]palmitic acid (PAL). The major phospholipid sources of the released [14C]AA were shown to be phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol. The participation of protein kinase C (PKC) in the enhanced synergistic release of [14C]AA was demonstrated by the inhibition of the release by the PKC inhibitor, staurosporine. Approximately 2-6% of the labeled AA liberated was converted to 5-hydroxyeicosatetraenoic acid by an endogenous 5-lipoxygenase. Therefore during cell activation the B cell is capable of liberating AA via a PKC-dependent mechanism, implicating AA and/or its metabolites in signal transduction.     

An orally available,brain-penetrant CAMKK2 inhibitor reduces food intake in rodent model

Hypothalamic CAMKK2 represents a potential mechanism for chemically affecting satiety and promoting weight loss in clinically obese patients. Single-digit nanomolar inhibitors of CAMKK2 were identified in three related ATP-competitive series. Limited optimization of kinase selectivity, solubility, and pharmacokinetic properties were undertaken on all three series, as SAR was often transferrable. Ultimately, a 2,4-diaryl 7-azaindole was optimized to afford a tool molecule that potently inhibits AMPK phosphorylation in a hypothalamus-derived cell line, is orally bioavailable, and crosses the blood–brain barrier. When dosed orally in rodents, compound 4?t limited ghrelin-induced food intake.     

Physiological methods to study biofilm disinfection

This report reviews the development of a rapidin situ approach to study the physiological responses of bacteria within biofilms to disinfectants. One method utilized direct viable counts (DVC) to assess the disinfection efficacy when thin biofilms were exposed to chlorine or monochloramine. Results obtained using the DVC method were one log higher than plate count (PC) estimates of the surviving population after disinfection. Other methods incorporated the use of fluorogenic stains, a cryotomy technique to yield thin (5-m) sections of biofilm communities and examination by fluorescence microscopy. The fluorogenic stains used in this approach included 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), which indicates cellular electron transport activity and Rhodamine 123, which responds specifically to proton motive force. The use of these stains allowed the microscopic discrimination of physiologically active bacteria as well as heterogeneities of active cells within thicker biofilms. The results of experiments using these techniques with pure culture and binary population biofilms on stainless steel coupons indicated biocidal activity of chlorine-based disinfectants occurred initially at the bulk-fluid interface of the communities and progressed toward the substratum. This approach provided a unique opportunity to describe the spatial response of bacteria within biofilms to antimicrobial agents and address mechanisms explaining their comparative resistance to disinfection in a way that has not been possible using traditional approaches. Results obtained using this alternative approach were also consistently higher than PC data following disinfection. These observations suggest that traditional methods involving biofilm removal and bacterial enumeration by colony formation overestimate biocide efficacy. Hence the alternative approach described here more accurately indicates the ability of bacteria surviving disinfection to recover and grow as well as demonstrate spatial heterogeneities in cellular physiological activities within biofilms.