Reliable and specific detection of Acanthamoeba spp. in dishcloths using quantitative real-time PCR assay.

Acanthamoeba spp., are ubiquitous protist which belongs to Free-Living Amoeba (FLA) group, is considered as causal agent of side-threatening keratitis or fatal encephalitis among other human infections. Besides, this parasite has been reported as host for other microorganisms important to human health such as Campylobacter spp. or Vibrio spp. among others. This role of Acanthamoeba as pathogen and environmental phagocyte has increased the reports confirming its presence in human related environments, acting as a water quality indicator. Considering the tide relationship between water and kitchen environments, and the high prevalence of Acanthamoeba in water sources, the present study aims to establish a quick and accurate protocol based on DNA extraction and a real time qPCR assay to detect Acanthamoeba spp. in dishcloths. The procedure has been validated by processing 17 used dishcloths. Our findings demonstrated the high sensitivity of the qPCR assay used which was capable of detecting up to one Acanthamoeba from an in vitro contaminated dishcloth. The protocol accurately detected 64.7% of positive samples for Acanthamoeba spp, (in 4 samples DNA concentrations corresponded to 1-102 amoebae). Our findings demonstrate the importance of FLA surveillance by efficient and sensitive methods since one amoeba is capable of colonizing human related food environments such as kitchens sinks and could be a potential source of infection.

Telomere length dynamics over 10-years and related outcomes in patients with COPD.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) has been proposed as a disease of accelerated aging. Several cross-sectional studies have related a shorter telomere length (TL), a marker of biological aging, with COPD outcomes. Whether accelerated telomere shortening over time relates to worse outcomes in COPD patients, is not known. METHODS: Relative telomere length (T/S) was determined by qPCR in DNA samples from peripheral blood in 263 patients at baseline and up to 10 years post enrolment. Yearly clinical and lung function data of 134 patients with at least two-time measures of T/S over this time were included in the analysis. RESULTS: At baseline, T/S inversely correlated with age (r = - 0.236; p < 0.001), but there was no relationship between T/S and clinical and lung function variables (p > 0.05). Over 10 years of observation, there was a median shortening of TL of 183 bp/year for COPD patients. After adjusting for age, gender, active smoking and mean T/S, patients that shortened their telomeres the most over time, had worse gas exchange, more lung hyperinflation and extrapulmonary affection during the follow-up, (PaO2 p < 0.0001; KCO p = 0.042; IC/TLC p < 0.0001; 6MWD p = 0.004 and BODE index p = 0.009). Patients in the lowest tertile of T/S through the follow-up period had an increased risk of death [HR = 5.48, (1.23-24.42) p = 0.026]. CONCLUSIONS: This prospective study shows an association between accelerated telomere shortening and progressive worsening of pulmonary gas exchange, lung hyperinflation and extrapulmonary affection in COPD patients. Moreover, persistently shorter telomeres over this observation time increase the risk for all-cause mortality.

C-reactive protein levels and clinically important predictive outcomes in stable COPD patients.

The aim of this study was to determine the relationship between C-reactive protein (CRP) levels and factors known to predict outcome in stable chronic obstructive pulmonary disease (COPD) patients. The following were studied in 130 stable COPD patients: spirometry, lung volume, arterial oxygen tension (P(a,O2)), dyspnoea, 6-min walk distance (6MWD), body mass index, fat-free mass index, BODE (body mass index, obstruction, dyspnoea and exercise capacity), health-related quality of life, smoking status, the presence of cardiovascular risk factors or disease, corticosteroid use and number of exacerbations in the previous year. CRP levels were measured in these patients and in 65 controls. Using univariate and multivariate analyses, any possible association with the predictors of outcomes was evaluated. CRP levels were higher in COPD patients than in controls (4.1 versus 1.8 mg.L(-1), respectively). Correlation was found with the following variables: forced expiratory volume in one second (FEV1; -0.23), FEV1 % (-0.20), forced vital capacity (FVC; -0.24), FVC % (-0.24), Global Initiative for Chronic Obstructive Lung Disease stage (0.17), BODE (0.17), inspiratory capacity/total lung capacity (-0.20), P(a,O2) (-0.40) and 6MWD (-0.30). Using multivariate analysis, P(a,O2) and 6MWD manifested the strongest negative association with CRP levels. C-reactive protein levels in stable chronic obstructive pulmonary disease patients are best correlated with arterial oxygen tension and 6-min walk distance. This should be considered when C-reactive protein levels are measured in stable chronic obstructive pulmonary disease patients.

[Tegumentary leishmaniasis in area with epidemic levels of transmission, Salta, Argentina, 1998]. / Leishmaniosis tegumentaria en un área con niveles epidémicos de transmisión, Salta, Argentina, 1998.

The incidence of tegumentary leishmaniasis increased in October 1997, in an area close to Oran city, province of Salta. The leishmaniasis' epidemiology and the entomology of this outbreak were studied during June 1998 at Paraje Las Carmelitas and Río Blanco, places with high reported incidence. The prevalence of infection (Montenegro Skin Test reactive) was 171/1000 inhabitants in Paraje Las Carmelitas and 790/1000 in Río Blanco, while the rates of leishmaniasis' incidence (active ulcers) were 72/1000 and 790/1000. The prevalence differences among sex and age groups were not significant. The clinical and epidemiologic patterns were consistent with those reported in the area for Leishmania (Viannia) braziliensis. Lutzomyia intermedia was the prevalent species (96%) among 2577 Phlebotominae collected, Lu. shannoni, Lu. cortelezzii y Lu. migonei were also present. The results about distribution of prevalence by age, and spatial distribution of Phlebotominae supported deforestation-residual primary forest transmission hypotheses. Therefore, in Salta the epidemic outbreaks generated by extensive deforestation could be sustained and amplified because the human settlements are located close to the remaining primary forest "spots". Furthermore, in this scenario the risk of human-vector contact increases by the subsistence or recreational activities performed in secondary transitional forests, and the presence of domestic animals around the houses. The results are discussed in the frame of possible control strategies.

Leishmaniosis tegumentaria en un área con niveles epidémicos de transmisión, Salta, Argentina, 1998. / [Tegumentary leishmaniasis in area with epidemic levels of transmission, Salta, Argentina, 1998]

The incidence of tegumentary leishmaniasis increased in October 1997, in an area close to Oran city, province of Salta. The leishmaniasis epidemiology and the entomology of this outbreak were studied during June 1998 at Paraje Las Carmelitas and Río Blanco, places with high reported incidence. The prevalence of infection (Montenegro Skin Test reactive) was 171/1000 inhabitants in Paraje Las Carmelitas and 790/1000 in Río Blanco, while the rates of leishmaniasis incidence (active ulcers) were 72/1000 and 790/1000. The prevalence differences among sex and age groups were not significant. The clinical and epidemiologic patterns were consistent with those reported in the area for Leishmania (Viannia) braziliensis. Lutzomyia intermedia was the prevalent species (96

) among 2577 Phlebotominae collected, Lu. shannoni, Lu. cortelezzii y Lu. migonei were also present. The results about distribution of prevalence by age, and spatial distribution of Phlebotominae supported deforestation-residual primary forest transmission hypotheses. Therefore, in Salta the epidemic outbreaks generated by extensive deforestation could be sustained and amplified because the human settlements are located close to the remaining primary forest [quot ]spots[quot ]. Furthermore, in this scenario the risk of human-vector contact increases by the subsistence or recreational activities performed in secondary transitional forests, and the presence of domestic animals around the houses. The results are discussed in the frame of possible control strategies.