OBJECTIVES: This study aimed to provide the first evidence of the socioeconomic burden of cystic fibrosis (CF) in Czechia. METHODS: In a cross-sectional questionnaire-based primary data collection conducted from 2020 to 2021 among Czech patients with CF, we collected demographic, clinical, and healthcare resource use data, out-of-pocket and social transfer costs, and questionnaires: Cystic Fibrosis Questionnaire-Revised, Work Productivity and Activity Impairment, EQ-5D, and Zarit Burden Interview. Productivity loss/costs were assessed using the human capital approach with patient patient-assumed life expectancy of 45 years and caregiver retirement age of 64 years and discounted by 3%. RESULTS: A total of 257 patients completed the questionnaires (37% of the Czech CF population). The average age was 17 years; most were females (59%), and the average forced expiratory volume in 1 second was 81.4% (SD 25.4%). A total of 107 patients had caregivers with an average age of 39 years and a significant caregiver time burden (extra 4.6 hours/day). The average Zarit Burden Interview score (25.4) was comparable with advanced cancer, dementia, or Duchenne muscular dystrophy. The proportion of unemployed caregivers was 10× higher than the general population (31% vs 3.2%). Total out-of-pocket family costs related to CF were €278/month, mainly for medicines (€105), foods (€73), and transport (€59); 25% received a disability pension and 18% other social security benefits. The work impairment of employed patients and caregivers was 25% and 15%, respectively, mostly due to presenteeism. Total lifetime productivity costs extrapolated to all Czech patients with CF (n = 687) and their caregivers were €155 181 286 (€225 883/person). CONCLUSIONS: The societal burden imposed on Czech patients with CF and their caregivers is significant. Caregivers seem to be affected by higher disease activity more than patients.
- Klíčová slova
- caregiver burden, cystic fibrosis, productivity costs, quality of life, societal burden,
- Publikační typ
- časopisecké články MeSH
The use of plant-derived natural products for the treatment of tropical parasitic diseases often has ethnopharmacological origins. As such, plants grown in temperate regions remain largely untested for novel anti-parasitic activities. We describe here a screen of the PhytoQuest Phytopure library, a novel source comprising over 600 purified compounds from temperate zone plants, against in vitro culture systems for Plasmodium falciparum, Leishmania mexicana, Trypanosoma evansi and T. brucei. Initial screen revealed 6, 65, 15 and 18 compounds, respectively, that decreased each parasite's growth by at least 50% at 1-2 µM concentration. These initial hits were validated in concentration-response assays against the parasite and the human HepG2 cell line, identifying hits with EC50 < 1 μM and a selectivity index of >10. Two sesquiterpene glycosides were identified against P. falciparum, four sterols against L. mexicana, and five compounds of various scaffolds against T. brucei and T. evansi. An L. mexicana resistant line was generated for the sterol 700022, which was found to have cross-resistance to the anti-leishmanial drug miltefosine as well as to the other leishmanicidal sterols. This study highlights the potential of a temperate plant secondary metabolites as a novel source of natural products against tropical parasitic diseases.
- Klíčová slova
- African trypanosomiasis, Surra, drug discovery, leishmaniasis, malaria, natural products, neglected tropical diseases, temperate zone,
- Publikační typ
- časopisecké články MeSH
Trichobilharzia regenti and T. szidati are schistosomes that infect birds. although T. regenti/T. szidati can only complete their life cycle in specific bird hosts (waterfowl), their larvae-cercariae are able to penetrate, transform and then migrate as schistosomula in nonspecific hosts (e.g., mouse, man). Peptidases are among the key molecules produced by these schistosomes that enable parasite invasion and survival within the host and include cysteine peptidases such as cathepsins B1 and B2. These enzymes are indispensable bio-catalysts in a number of basal biological processes and host-parasite interactions, e.g., tissue invasion/migration, nutrition and immune evasion. Similar biochemical and functional characteristics were observed for cathepsins B1 and B2 in bird schistosomes (T. regenti, T. szidati) and also for their homologs in human schistosomes (Schistosoma mansoni, S. japonicum). Therefore, data obtained in the research of bird schistosomes can also be exploited for the control of human schistosomes such as the search for targets of novel chemotherapeutic drugs and vaccines.
- MeSH
- cerkárie enzymologie MeSH
- kathepsiny chemie genetika metabolismus MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- nemoci ptáků parazitologie MeSH
- ptáci parazitologie MeSH
- Schistosoma enzymologie MeSH
- sekvence aminokyselin MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- kathepsiny MeSH
The neurotropic bird schistosome Trichobilharzia regenti possesses papain-like cysteine peptidases which have also been shown to be crucial enzymes in various developmental stages of the related human parasites Schistosoma spp. In this paper, we present data obtained by real-time polymerase chain reaction on the temporal distribution of transcripts of two cathepsins in different developmental stages of T. regenti: cathepsin B1 originally described from the gut lumen of schistosomula with presumptive role in nutrient digestion and cathepsin B2 originally found in penetration glands of cercariae with probable involvement in invasion of the final host. In spite of their mutual resemblance at the sequence level, the mRNA expression profiles clearly show distinct expression of cathepsins B1 and B2 during the development from eggs to cercariae. In the case of both cathepsins, the highest level of transcription was detected in intravertebrate stages. Putative functions of cathepsins B1 and B2 in schistosome developmental stages are discussed.
- MeSH
- hlemýždi MeSH
- izoenzymy genetika metabolismus MeSH
- kachny MeSH
- kathepsin B genetika metabolismus MeSH
- proteiny červů genetika metabolismus MeSH
- Schistosomatidae enzymologie genetika růst a vývoj MeSH
- stadia vývoje MeSH
- stanovení celkové genové exprese * MeSH
- vývojová regulace genové exprese * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- izoenzymy MeSH
- kathepsin B MeSH
- proteiny červů MeSH
A transcriptional product of a gene encoding cathepsin B-like peptidase in the bird schistosome Trichobilharzia regenti was identified and cloned. The enzyme was named TrCB2 due to its 77% sequence similarity to cathepsin B2 from the important human parasite Schistosoma mansoni. The zymogen was expressed in the methylotropic yeast Pichia pastoris; procathepsin B2 underwent self-processing in yeast media. The peptidolytic activity of the recombinant enzyme was characterised using synthetic fluorogenic peptide substrates at optimal pH 6.0. Functional studies using different specific inhibitors proved the typical cathepsin B-like nature of the enzyme. The S(2) subsite specificity profile of recombinant TrCB2 was obtained. Using monospecific antibodies against the recombinant enzyme, the presence of cathepsin B2 was confirmed in extracts from cercariae (infective stage) and schistosomula (early post-cercarial stage) of T. regenti on Western blots. Also, cross-reactivity was observed between T. regenti and S. mansoni cathepsins B2 in extracts of cercariae, schistosomula or adults. In T. regenti, the antisera localised the enzyme to post-acetabular penetration glands of cercariae implying an important role in the penetration of host skin. The ability of recombinant TrCB2 to degrade skin, serum and nervous tissue proteins was evident. Elastinolytic activity suggests that the enzyme might functionally substitute the histolytic role of the serine class elastase known from S. mansoni and Schistosoma haematobium but not found in Schistosoma japonicum or in bird schistosomes.
- MeSH
- cystein biosyntéza MeSH
- cysteinové proteasy biosyntéza genetika fyziologie MeSH
- hlemýždi MeSH
- kachny MeSH
- kathepsin B biosyntéza MeSH
- králíci MeSH
- krocani MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- Pichia MeSH
- Schistosoma mansoni enzymologie MeSH
- Schistosoma enzymologie MeSH
- sekvence aminokyselin MeSH
- skot MeSH
- substrátová specifita MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- lidé MeSH
- myši MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- cystein MeSH
- cysteinové proteasy MeSH
- kathepsin B MeSH
Trichobilharzia regenti is a neurotropic bird schistosome,causing cercarial dermatitis in humans. In this study, ZAP cDNA expression library from Radix peregra s. lat. hepatopancreases containing intramolluscan stages of T. regenti was constructed and screened using PCR with specific and degenerate primers, designed according to previously described serine and cysteine peptidases of other parasite species. Full-length sequences of cathepsins B1 and L, and two serine peptidases, named RpSP1 and RpSP2, were obtained. The protein-protein BLAST analysis and parallel control reactions with template from hepatopancreases of T. regenti non-infected snails revealed that only cathepsin B1 was of parasite origin. The remaining sequences were derived from the snail intermediate host, which implies that the initial source of parasite mRNA was contaminated by snail tissue. Regardless of this contamination, the cDNA library remains an excellent molecular tool for detection and identification of bioactive molecules in T. regenti cercariae.
- MeSH
- cysteinové endopeptidasy genetika MeSH
- genová knihovna MeSH
- hlemýždi parazitologie MeSH
- kathepsin B genetika MeSH
- kathepsin L MeSH
- kathepsiny genetika MeSH
- molekulární sekvence - údaje MeSH
- polymerázová řetězová reakce MeSH
- proteasy genetika MeSH
- proteiny červů genetika MeSH
- Schistosomatidae enzymologie genetika růst a vývoj MeSH
- sekvenční analýza DNA MeSH
- sekvenční homologie aminokyselin MeSH
- stadia vývoje MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- CTSL protein, human MeSH Prohlížeč
- cysteinové endopeptidasy MeSH
- kathepsin B MeSH
- kathepsin L MeSH
- kathepsiny MeSH
- proteasy MeSH
- proteiny červů MeSH
