The taeniid tapeworm Echinococcus granulosus is the causative agent of echinococcal disease, a major zoonosis with worldwide distribution. Several efforts to establish an in vitro model of E. granulosus have been undertaken; however, many of them have been designed for Echinococcus multilocularis. In the present study, we have described and characterized a stable cell line obtained from E. granulosus bovine protoscoleces maintained 3 yr in vitro. Growth characterization, morphology by light, fluorescent and electronic microscopy, and karyotyping were carried out. Cell culture origin was confirmed by immunofluorescent detection of AgB4 antigen and by PCR for the mitochondrial cytochrome c-oxidase subunit 1 (DCO1) gene. Cells seeded in agarose biphasic culture resembled a cystic structure, similar to the one formed in secondary hosts. This cell line could be a useful tool to research equinococcal behavior, allowing additional physiological and pharmacological studies, such as the effect of growth factors, nutrients, and antiparasitic drugs on cell viability and growth and on cyst formation.
Bisphosphonates have been proposed as pharmacological agents against parasite and cancer cell growth. The effect of these compounds on helminthic cell viability and acellular compartment morphology, however, has not yet been studied. The effects of different types of bisphosphonates, namely etidronate (EHDP), pamidronate (APD), alendronate (ABP), ibandronate (IB) and olpadronate (OPD), and their interaction with amiloride, 1,25-dihydroxycholecalciferol (D3) and proline were evaluated on a cell line derived from bovine Echinococcus granulousus protoscoleces (EGPE) that forms cystic colonies in agarose. The EGPE cell line allowed testing the effect of bisphosphonates alone and in association with other compounds that could modulate calcium apposition/deposition, and were useful in measuring the impact of these compounds on cell growth, cystic colony formation and calcium storage. Decreased cell growth and cystic colony formation were found with EHDP, IB and OPD, and increased calcium storage with EHDP only. Calcium storage in EGPE cells appeared to be sensitive to the effect of amiloride, D3 and proline. Proline decreased calcium storage and increased colony formation. Changes in calcium storage may be associated with degenerative changes of the cysts, as shown in the in vitro colony model and linked to an adenosine triphosphate (ATP) decrease. In conclusion, bisphosphonates could be suitable tempering drugs to treat cestode infections.
Introduction Coronavirus disease 2019 (COVID-19) is a highly pathogenic coronavirus characterized by by systemic inflammatory response with endothelial damage and a dysregulated coagulation system. Despite most patients survive the acute setting of COVID-19, their long-term clinical sequelae are highly unclear. We have sought to identify the impact of post-COVID-19 syndrome on mid-term follow-up and gain some additional insights about the potential explanation for persistence of dyspnea. Methods This is a 3-month prospective cohort study of previously hospitalised COVID-19 patients recruited from a single Spanish center, a small outpatient group without prior hospitalisation was also evaluated. Patients underwent serial testing with cardio-pulmonary exercise test (CPET), transthoracic echocardiogram, pulmonary lung test, six-minute walking test, serum biomarker analysis and quality of life questionaries. They were classify according to the presence of persistent dyspnea. Primary study outcome was predicted peak oxygen consumption (V02) according to CPET and predicted carbon monoxide diffusion capacity. Results Our study included 41 (58.6%) patients with dyspnea and 29 (41.4%) asymptomatic. Symptomatic patients had a higher proportion of females (73.2% vs. 51.7%), but comparable age and prevalence of cardiovascular risk factors. We did not observe differences among the assessed variables in transthoracic echocardiogram and pulmonary function test. Patients who referred dyspnea had smaller predicted peak O2 consumption (77.8 [64–92.5] vs. 99 [88–105]: p<0.001), total distance in the 6-minute walking test (535 [467–600] vs. 611 [550–650] meters; p=0.001), and quality of life (KCCQ-23 60.1±18.6 vs. 82.8±11.3; p<0.001). Additionally, abnormalities in CPET were suggestive of a ventilation/perfusion misthmach or hyperventilatory syndrome characterized by impaired ventilatory efficiency with a greater VE/VCO2 slope (32 [28.1–37.4] vs. 29.4 [26.9–31.4]; p=0.022) and low PETCO2 (34 [32–39] vs. 38 [36–40]; p=0.025). Interpretation In this study >50% of COVID-19 survivors present a symptomatic functional impairment irrespective of age or prior hospitalization. Compared to asymptomatic patients, among those who referred dyspnea our findings suggest potential ventilatory inefficiency. FUNDunding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Gerencia Regional de Salud de Castilla y Leόn; Grant from the Spanish Society of Cardiology Tabla de resultados
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