A False Start in the Race Against Doping in Sport: Concerns With Cycling’s Biological Passport

Professional cycling has suffered from a number of doping scandals. The sport’s governing bodies have responded by implementing an aggressive new antidoping program known as the biological passport. Cycling’s biological passport marks a departure from traditional antidoping efforts, which have focused on directly detecting prohibited substances in a cyclist’s system. Instead, the biological passport tracks biological variables in a cyclist’s blood and urine over time, monitoring for fluctuations that are thought to indirectly reveal the effects of doping. Although this method of indirect detection is promising, it also raises serious legal and scientific concerns. Since its introduction, the cycling community has debated the reliability of indirect biological-passport evidence and the clarity, consistency, and transparency of its use in proving doping violations. Such uncertainty undermines the legitimacy of finding cyclists guilty of doping based on this indirect evidence alone. Antidoping authorities should address these important concerns before continuing to pursue doping sanctions against cyclists solely on the basis of their biological passports

Establishment of a Novel Fluorescence-Based Method to Evaluate Chaperone-Mediated Autophagy in a Single Neuron

Background: Chaperone-mediated autophagy (CMA) is a selective autophagy-lysosome protein degradation pathway. The role of CMA in normal neuronal functions and in neural disease pathogenesis remains unclear, in part because there is no available method to monitor CMA activity at the single-cell level. Methodology/Principal Findings: We sought to establish a single-cell monitoring method by visualizing translocation of CMA substrates from the cytosol to lysosomes using the HaloTag (HT) system. GAPDH, a CMA substrate, was fused to HT (GAPDH-HT); this protein accumulated in the lysosomes of HeLa cells and cultured cerebellar Purkinje cells (PCs) after labeling with fluorescent dye-conjugated HT ligand. Lysosomal accumulation was enhanced by treatments that activate CMA and prevented by siRNA-mediated knockdown of LAMP2A, a lysosomal receptor for CMA, and by treatments that inactivate CMA. These results suggest that lysosomal accumulation of GAPDH-HT reflects CMA activity. Using this method, we revealed that mutant cPKC, which causes spinocerebellar ataxia type 14, decreased CMA activity in cultured PCs. Conclusion/Significance: In the present study, we established a novel fluorescent-based method to evaluate CMA activity in a single neuron. This novel method should be useful and valuable for evaluating the role of CMA in various neurona

Tissue Invasion by Entamoeba histolytica: Evidence of Genetic Selection and/or DNA Reorganization Events in Organ Tropism

Entamoeba histolytica infection may have various clinical manifestations. Nine out of ten E. histolytica infections remain asymptomatic, while the remainder become invasive and cause disease. The most common form of invasive infection is amebic diarrhea and colitis, whereas the most common extra-intestinal disease is amebic liver abscess. The underlying reasons for the different outcomes are unclear, but a recent study has shown that the parasite genotype is a contributor. To investigate this link further we have examined the genotypes of E. histolytica in stool- and liver abscess-derived samples from the same patients. Analysis of all 18 paired samples (16 from Bangladesh, one from the United States of America, and one from Italy) revealed that the intestinal and liver abscess amebae are genetically distinct. The results suggest either that E. histolytica subpopulations in the same infection show varying organ tropism, or that a DNA reorganization event takes place prior to or during metastasis from intestine to liver

Modeling the extrinsic incubation of Dirofilaria immitis in South America based on monthly and continuous climatic data

The spread and recrudescence of dirofilariosis across several regions, either caused by Dirofilaria immitis or D. repens, responds to many factors. Building upon the temperature model by Slocombe et al. (1989), a number of studies have been performed to generate predictive models for dirofilariosis. These studies have demonstrated the length of transmission periods and the number of Dirofilaria generations supported throughout several regions of the world (either at large or at small-scale). The usual procedure have proved to be extremely time consuming, as it appears impractical when assessing potential transmission at large scale, such as at a country or large-scale level. Due to the increasing need to suggest standardized surveillance protocols and apply adequate preventive measures at national and regional levels, a model for calculation of Dirofilaria HDUs based on monthly data was adapted for large-scale regional use. The models proposed are based on both point data (meteorological stations) and interpolated climate data layers (WorldClim). Three different models (daily and monthly models based on point data, and monthly model based on continuous data) were developed and compared statistically. When compared with the results from the classical daily model, the monthly models proposed accurately predicted the locations were extrinsic incubation was possible. These models proved to be adequate for the regional analysis of the extrinsic incubation of D. immitis and, hence, the relative risk of transmission in South America. Further, these models confirm that favorable temperatures for heartworm transmission in South America are present in most of the countries. D. immitis extrinsic incubation follows a seasonal pattern in Argentina, Chile, Uruguay, eastern Paraguay and southeastern Brazil; while in northern half of South America (less than 25° S) transmission may occur year-round. Moreover, high risk areas suitable for dirofilariasis transmission are not geographically constant throughout the year. The validation procedures indicate that the predicted HDU and HG maps are good predictors of dirofilariosis potential distribution, but estimating dirofilariosis prevalences based on them might not be completely accurate. The resulting distribution and seasonal maps would be useful for heartworm prevention by chemoprophylaxis in different regions known to be endemic for canine dirofilariasis. The information here provided can be an important tool in veterinary public health, as well as a guide for future research

Analysis of climate and extrinsic incubation of Dirofilaria immitis in southern South America.

Dirofilariosis, caused by Dirofilaria immitis and D. repens, is spreading in several geographic regions. The deve - lopment of infective larvae in the mosquito vector (extrinsic incubation) needs an accumulated total of 130 degree-days above the 14 ºC threshold, normally expressed as heartworm development units (HDUs). Based on this information, temperature- based models have been developed and applied to evaluate the distribution and spread of Dirofilaria infections in various countries and continents. Despite the confirmed presence of D. immitis in most South American countries, the available information about its epidemiology remains scarce. We analysed the temporal and spatial extrinsic incubation of this parasite in Argentina, Chile and Uruguay, taking into account daily temperatures from 49 meteorological stations during a 30-year period (1982-2012). The theoretically possible number of D. immitis generations was calculated based on the number of meteorological stations that reached the 130-HDUs threshold. The resulting information was spatially interpolated using the inverse weighted distance (IWD) model to produce thematic maps. The model shows that 41 of the meteorological stations reach the threshold needed and that D. immitis transmission is markedly seasonal with a peak in late spring (December), stable during summer (January to March) and declining in the autumn (April and May). Suitable temperatures exist in Uruguay and most of Argentina, whereas D. immitis transmission in Chile is only possible in the north and in the central inlands. The results suggest that the climatic impact on D. immitis transmission must have been minimal in the countries investigated since the annual meteorological records did not change much during the 30-year period analysed

Detection of Dirofilaria immitis in mid-western arid Argentina.

Dirofilariosis, caused by Dirofilaria immitis and D. repens, is (re-) emerging worldwide. Dogs are the main reservoirs, while human infection has recently become an important focus of interest and attention. In Argentina, canine D. immitis infection has been described in eastern and northern subtropical and temperate humid regions, but never reported in mid-western arid regions so far. In this research note we report for the first time the occurrence of autochthonous human and canine D. immitis infection in the region

Chaperone-mediated autophagy: roles in disease and aging

This review focuses on chaperone-mediated autophagy (CMA), one of the proteolytic systems that contributes to degradation of intracellular proteins in lysosomes. CMA substrate proteins are selectively targeted to lysosomes and translocated into the lysosomal lumen through the coordinated action of chaperones located at both sides of the membrane and a dedicated protein translocation complex. The selectivity of CMA permits timed degradation of specific proteins with regulatory purposes supporting a modulatory role for CMA in enzymatic metabolic processes and subsets of the cellular transcriptional program. In addition, CMA contributes to cellular quality control through the removal of damaged or malfunctioning proteins. Here, we describe recent advances in the understanding of the molecular dynamics, regulation and physiology of CMA, and discuss the evidence in support of the contribution of CMA dysfunction to severe human disorders such as neurodegeneration and cancer