(1) Background: Training load monitoring has become a relevant research-practice gap to control training and match demands in team sports. However, there are no systematic reviews about accumulated training and match load in football. (2) Methods: Following the preferred reporting item for systematic reviews and meta-analyses (PRISMA), a systematic search of relevant English-language articles was performed from earliest record to March 2020. The search included descriptors relevant to football, training load, and periodization. (3) Results: The literature search returned 7972 articles (WoS = 1204; Pub-Med = 869, SCOPUS = 5083, and SportDiscus = 816). After screening, 36 full-text articles met the inclusion criteria and were reviewed. Eleven of the included articles analyzed weekly training load distribution; fourteen, the weekly training load and match load distribution; and eleven were about internal and external load relationships during training. The reviewed articles were based on short-telemetry systems (n = 12), global positioning tracking systems (n = 25), local position measurement systems (n = 3), and multiple-camera systems (n = 3). External load measures were quantified with distance and covered distance in different speed zones (n = 27), acceleration and deceleration (n = 13) thresholds, accelerometer metrics (n = 11), metabolic power output (n = 4), and ratios/scores (n = 6). Additionally, the internal load measures were reported with perceived exertion (n = 16); heart-rate-based measures were reported in twelve studies (n = 12). (4) Conclusions: The weekly microcycle presented a high loading variation and a limited variation across a competitive season. The magnitude of loading variation seems to be influenced by the type of week, player’s starting status, playing positions, age group, training mode and contextual variables. The literature has focused mainly on professional men; future research should be on the youth and female accumulated training/match load monitoring.

Cryptosporidiosis causes life-threatening diarrhea in children under the age of 5 years and prolonged diarrhea in immunodeficient people, especially AIDS patients. The standard of care, nitazoxanide, is modestly effective in children and ineffective in immunocompromised individuals. In addition to the need for new drugs, better knowledge of drug properties that drive in vivo efficacy is needed to facilitate drug development. We report the identification of a piperazine-based lead compound for Cryptosporidium drug development, MMV665917, and a new pharmacodynamic method used for its characterization. The identification of MMV665917 from the Medicines for Malaria Venture Malaria Box was followed by dose-response studies, in vitro toxicity studies, and structure-activity relationship studies using commercial analogues. The potency of this compound against Cryptosporidium parvum Iowa and field isolates was comparable to that against Cryptosporidium hominis. Furthermore, unlike nitazoxanide, clofazimine, and paromomycin, MMV665917 appeared to be curative in a NOD SCID gamma mouse model of chronic cryptosporidiosis. MMV665917 was also efficacious in a gamma interferon knockout mouse model of acute cryptosporidiosis. To determine if efficacy in this mouse model of chronic infection might relate to whether compounds are parasiticidal or parasitistatic for C. parvum, we developed a novel in vitro parasite persistence assay. This assay suggested that MMV665917 was parasiticidal, unlike nitazoxanide, clofazimine, and paromomycin. The assay also enabled determination of the concentration of the compound required to maximize the rate of parasite elimination. This time-kill assay can be used to prioritize early-stage Cryptosporidium drug leads and may aid in planning in vivo efficacy experiments. Collectively, these results identify MMV665917 as a promising lead and establish a new method for characterizing potential anticryptosporidial agents.

Cryptosporidiosis is a leading cause of life-threatening diarrhea in children, and the only currently approved drug is ineffective in malnourished children and immunocompromised people. Large-scale phenotypic screens are ongoing to identify anticryptosporidial compounds, but optimal approaches to prioritize inhibitors and establish a mechanistically diverse drug development pipeline are unknown. Here, we present a panel of medium-throughput mode of action assays that enable testing of compounds in several stages of the Cryptosporidium life cycle. Phenotypic profiles are given for thirty-nine anticryptosporidials. Using a clustering algorithm, the compounds sort by phenotypic profile into distinct groups of inhibitors that are either chemical analogs (i.e. same molecular mechanism of action (MMOA)) or known to have similar MMOA. Furthermore, compounds belonging to multiple phenotypic clusters are efficacious in a chronic mouse model of cryptosporidiosis. This suite of phenotypic assays should ensure a drug development pipeline with diverse MMOA without the need to identify underlying mechanisms.

Monitoring the training load in football is an important strategy to improve athletic performance and an effective training periodization. The aim of this study was two-fold: (1) to quantify the weekly training load and recovery status variations performed by under-15, under-17 and under-19 sub-elite young football players; and (2) to analyze the influence of age, training day, weekly microcycle, training and playing position on the training load and recovery status. Twenty under-15, twenty under-17 and twenty under-19 players were monitored over a 2-week period during the first month of the 2019–2020 competitive season. Global positioning system technology (GPS) was used to collect external training loads: total distance covered, average speed, maximal running speed, relative high-speed running distance, high metabolic load distance, sprinting distance, dynamic stress load, accelerations and decelerations. Internal training load was monitored using ratings of perceived exertion (RPE) and session rating of perceived exertion (sRPE). Recovery status was obtained using the total quality recovery (TQR) scale. The results show an age-related influence for external training load (p ≤ 0.001; d = 0.29–0.86; moderate to strong effect), internal training load (p ≤ 0.001, d = 0.12–0.69; minimum to strong effect) and recovery status (p ≤ 0.001, d = 0.59; strong effect). The external training load presented differences between training days (p < 0.05, d = 0.26–0.95; moderate to strong effect). The playing position had a minimum effect on the weekly training load (p < 0.05; d = 0.06–0.18). The weekly microcycle had a moderate effect in the TD (p < 0.05, d = 0.39), RPE (p < 0.05; d = 0.35) and sRPE (p < 0.05, d = 0.35). Interaction effects were found between the four factors analyzed for deceleration (F = 2.819, p = 0.017) and between inter-day, inter-week and age for total covered distance (F = 8.342, p = 0.008). This study provided specific insights about sub-elite youth football training load and recovery status to monitor training environments and load variations. Future research should include a longer monitoring period to assess training load and recovery variations across different season phases.

Entamoeba histolytica is an intestinal ameba that causes dysentery and liver abscesses. Cytotoxicity and phagocytosis of host cells characterize invasive E. histolytica infection. Prior to phagocytosis of host cells, E. histolytica induces apoptotic host cell death, using a mechanism that requires contact via an amebic galactosespecific lectin. However, lectin inhibition only partially blocks phagocytosis of already dead cells, implicating at least one additional receptor in phagocytosis. To identify receptors for engulfment of apoptotic cells, monoclonal antibodies against E. histolytica membrane antigens were screened for inhibition of phagocytosis. Of 43 antibodies screened, one blocked lectin-independent uptake of apoptotic cells, with >90% inhibition at a dose of 20 g/ml (P < 0.0003 versus control). The same antibody also inhibited adherence to apoptotic lymphocytes and, to a lesser extent, adherence to and killing of viable lymphocytes. The antigen recognized by the inhibitory antibody was purified by affinity chromatography and identified by liquid chromatography-mass spectrometry as the serine-rich E. histolytica protein (SREHP). Consistent with this, the inhibitory antibody bound to recombinant SREHP present in bacterial lysates on immunoblots. The SREHP is an abundant immunogenic surface protein of unclear function. The results of this unbiased antibody screen strongly implicate the SREHP as a participant in E. histolytica phagocytosis and suggest that it may play an important role in adherence to apoptotic cells.

ABSTRACTInvasive amebiasis due toEntamoeba histolyticainfection is an important cause of morbidity in developing countries. TheE. histolyticagenome contains two homologues to the metalloprotease leishmanolysin gene,Entamoeba histolytica MSP-1(EhMSP-1) andEhMSP-2, while the commensal amebaEntamoeba disparhas lostEhMSP-1. In this study, we sought to characterizeE. histolyticametallosurface protease 1 (EhMSP-1). Using immunoprecipitation and a model substrate, we found that EhMSP-1 was a functional metalloprotease. Confocal microscopy and flow cytometry revealed that EhMSP-1 localized to the cell surface and revealed the existence of distinct, nonclonal trophozoite populations with high and low EhMSP-1 surface abundance that became synchronized following serum starvation. Phenotypic assays were performed after silencingEhMSP-1. Adherence of EhMSP-1-deficient trophozoites to tissue culture cell monolayers was more than five times greater than that of control amebas, but surface staining of several antigens, including the galactose adherence lectin, was unchanged.EhMSP-1silencing similarly increased adherence to both viable and apoptotic Jurkat lymphocytes. Tissue culture cell monolayer destruction was reduced byEhMSP-1silencing, although it was blocked almost completely by inhibiting cysteine proteases. Consistent with a primary defect in regulation of amebic adherence,EhMSP-1silencing also resulted in reduced mobility on tissue culture cell monolayers and in increased phagocytosis. In conclusion, EhMSP-1 was shown to be a surface metalloprotease involved in regulation of amebic adherence, with additional effects on cell motility, cell monolayer destruction, and phagocytosis.

ABSTRACTPhagocytosis of host cells is characteristic of tissue invasion by the intestinal amebaEntamoeba histolytica, which causes amebic dysentery and liver abscesses.Entamoeba histolyticainduces host cell apoptosis and uses ligands, including C1q, on apoptotic cells to engulf them. Two mass spectrometry analyses identified calreticulin in amebic phagosome preparations, and, in addition to its function as an endoplasmic reticulum chaperone, calreticulin is believed to be the macrophage receptor for C1q. The purpose of this study was to determine if calreticulin functions as anE. histolyticaC1q receptor during phagocytosis of host cells. Calreticulin was localized to the surface ofE. histolyticaduring interaction with both Jurkat lymphocytes and erythrocytes and was present in over 75% of phagocytic cups during amebic erythrophagocytosis. Presence of calreticulin on the cell surface was further demonstrated using a method that selectively biotinylated cell surface proteins and by flow cytometry using trophozoites overexpressing epitope-tagged calreticulin. Regulated overexpression of calreticulin increasedE. histolytica's ability to phagocytose apoptotic lymphocytes and calcium ionophore-treated erythrocytes but had no effect on amebic adherence to or destruction of cell monolayers or surface expression of the GalNAc lectin and serine-richE. histolyticaprotein (SREHP) receptors. Finally,E. histolyticacalreticulin bound specifically to apoptotic lymphocytes and to human C1q. Collectively, these data implicate cell surface calreticulin as a receptor for C1q duringE. histolyticaphagocytosis of host cells.

Entamoeba histolytica, the cause of amebiasis, is believed to have no continuous endoplasmic reticulum (ER), with ER functions occurring in vesicles. Here, using an ER-targeted green fluorescent protein fusion protein and fluorescence loss in photobleaching, we have unambiguously demonstrated the presence of a continuous ER compartment in living E. histolytica trophozoites.