IPES
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OBJECTIVE: In order to introduce a complex system of monitoring and evaluation of the heath-related quality of life (HRQoL) of children and adolescents with epilepsy (CWE) and their families in the Czech Republic, we aimed to validate the Czech versions of two appropriate instruments - the Impact of Pediatric Epilepsy Scale (IPES) and the Quality of Life in Epilepsy Inventory for Adolescents (QOLIE-AD-48). METHODS: The verification of the 11-item IPES was carried out in the group of parents of 248 CWE aged 2-18 years. One hundred and thirty-five CWE from the given group aged 11-18 years then completed the 48-item QOLIE-AD-48. Internal consistency, test-retest reliability (with a three-month interval) and the factorial structure of the Czech versions were determined and compared with the original instruments. RESULTS: We found that the Czech version of the IPES exhibited very good psychometric properties including high internal consistency (Cronbach's alpha, α, of 0.93), high test-retest reliability (intraclass correlation coefficient, ICC, of 0.76) and the same 3-factor structure as the original instrument. The superiority of this 3-factor solution over the alternate 2-factor model proposed for some language versions of the IPES was determined using confirmatory factor analysis. We found 8 items in the Czech version of the QOLIE-AD-48 belonging to original Attitudes towards epilepsy and Social support subscales that do not fit well with the Czech version due to their low correlation with the total score and insufficient test-retest reliability and should be omitted. For the remaining 40 items, we have determined high internal consistency (α = 0.95) and test-retest reliability (ICC = 0.82). Confirmatory factor analysis revealed that the 6-factor solution derived from the original instrument (without two removed subscales) was appropriate for the Czech version. The individual subscales exhibited high internal consistency with α = 0.61-0.91. The external validation of both instruments was confirmed based on a significant correlation between test results and physicians' reports of the characteristics of the child's epilepsy. CONCLUSIONS: The Czech versions of both instruments studied are reliable and valid, and can be used in the next research focusing on the effect of different treatment approaches on the HRQoL of CWE and their families.
- MeSH
- dítě MeSH
- doxorubicin analogy a deriváty MeSH
- epilepsie * diagnóza MeSH
- jazyk (prostředek komunikace) MeSH
- kvalita života * MeSH
- lidé MeSH
- mladiství MeSH
- předškolní dítě MeSH
- průzkumy a dotazníky MeSH
- psychometrie MeSH
- reprodukovatelnost výsledků MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- předškolní dítě MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Alexander disease (AxD) is a rare and severe neurodegenerative disorder caused by mutations in glial fibrillary acidic protein (GFAP). While the exact disease mechanism remains unknown, previous studies suggest that mutant GFAP influences many cellular processes, including cytoskeleton stability, mechanosensing, metabolism, and proteasome function. While most studies have primarily focused on GFAP-expressing astrocytes, GFAP is also expressed by radial glia and neural progenitor cells, prompting questions about the impact of GFAP mutations on central nervous system (CNS) development. In this study, we observed impaired differentiation of astrocytes and neurons in co-cultures of astrocytes and neurons, as well as in neural organoids, both generated from AxD patient-derived induced pluripotent stem (iPS) cells with a GFAPR239C mutation. Leveraging single-cell RNA sequencing (scRNA-seq), we identified distinct cell populations and transcriptomic differences between the mutant GFAP cultures and a corrected isogenic control. These findings were supported by results obtained with immunocytochemistry and proteomics. In co-cultures, the GFAPR239C mutation resulted in an increased abundance of immature cells, while in unguided neural organoids and cortical organoids, we observed altered lineage commitment and reduced abundance of astrocytes. Gene expression analysis revealed increased stress susceptibility, cytoskeletal abnormalities, and altered extracellular matrix and cell-cell communication patterns in the AxD cultures, which also exhibited higher cell death after stress. Overall, our results point to altered cell differentiation in AxD patient-derived iPS-cell models, opening new avenues for AxD research.
- MeSH
- Alexanderova nemoc * genetika patologie metabolismus MeSH
- astrocyty * metabolismus patologie MeSH
- buněčná diferenciace * fyziologie MeSH
- gliový fibrilární kyselý protein * metabolismus genetika MeSH
- indukované pluripotentní kmenové buňky * metabolismus MeSH
- kokultivační techniky MeSH
- kultivované buňky MeSH
- lidé MeSH
- mutace MeSH
- nervové kmenové buňky metabolismus MeSH
- neurony metabolismus patologie MeSH
- organoidy metabolismus patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Spinal cord injury (SCI), is a devastating condition leading to the loss of locomotor and sensory function below the injured segment. Despite some progress in acute SCI treatment using stem cells and biomaterials, chronic SCI remains to be addressed. We have assessed the use of laminin-coated hydrogel with dual porosity, seeded with induced pluripotent stem cell-derived neural progenitors (iPSC-NPs), in a rat model of chronic SCI. iPSC-NPs cultured for 3 weeks in hydrogel in vitro were positive for nestin, glial fibrillary acidic protein (GFAP) and microtubule-associated protein 2 (MAP2). These cell-polymer constructs were implanted into a balloon compression lesion, 5 weeks after lesion induction. Animals were behaviorally tested, and spinal cord tissue was immunohistochemically analyzed 28 weeks after SCI. The implanted iPSC-NPs survived in the scaffold for the entire experimental period. Host axons, astrocytes and blood vessels grew into the implant and an increased sprouting of host TH+ fibers was observed in the lesion vicinity. The implantation of iPSC-NP-LHM cell-polymer construct into the chronic SCI led to the integration of material into the injured spinal cord, reduced cavitation and supported the iPSC-NPs survival, but did not result in a statistically significant improvement of locomotor recovery.
- MeSH
- buněčná diferenciace MeSH
- chronická nemoc MeSH
- hydrogely MeSH
- indukované pluripotentní kmenové buňky metabolismus MeSH
- krysa rodu rattus MeSH
- nervové kmenové buňky transplantace MeSH
- poranění míchy terapie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tick-borne encephalitis virus (TBEV) targets the central nervous system (CNS), leading to potentially severe neurological complications. The neurovascular unit plays a fundamental role in the CNS and in the neuroinvasion of TBEV. However, the role of human brain pericytes, a key component of the neurovascular unit, during TBEV infection has not yet been elucidated. In this study, TBEV infection of the primary human brain perivascular pericytes was investigated with highly virulent Hypr strain and mildly virulent Neudoerfl strain. We used Luminex assay to measure cytokines/chemokines and growth factors. Both viral strains showed comparable replication kinetics, peaking at 3 days post infection (dpi). Intracellular viral RNA copies peaked at 6 dpi for Hypr and 3 dpi for Neudoerfl cultures. According to immunofluorescence staining, only small proportion of pericytes were infected (3% for Hypr and 2% for Neudoerfl), and no cytopathic effect was observed in the infected cells. In cell culture supernatants, IL-6 production was detected at 3 dpi, together with slight increases in IL-15 and IL-4, but IP-10, RANTES and MCP-1 were the main chemokines released after TBEV infection. These chemokines play key roles in both immune defense and immunopathology during TBE. This study suggests that pericytes are an important source of these signaling molecules during TBEV infection in the brain.
- MeSH
- chemokin CCL5 * metabolismus MeSH
- chemokin CXCL10 * metabolismus MeSH
- cytokiny metabolismus MeSH
- klíšťová encefalitida * virologie metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- mozek * virologie metabolismus patologie MeSH
- pericyty * virologie metabolismus MeSH
- replikace viru MeSH
- viry klíšťové encefalitidy * fyziologie patogenita MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
OBJECTIVES: This study aimed to determine the occurrence of intra-amniotic inflammatory changes associated with chronic inflammation in the placenta, marked by elevated levels of interferon gamma-induced protein 10 (IP-10) (≥2200 pg/mL) in the amniotic fluid of women with preterm prelabor rupture of membranes (PPROM). Specifically, the study investigated whether these intra-amniotic inflammatory changes were more common in women with microbial invasion of amniotic cavity (MIAC) and intra-amniotic inflammation (IAI), as indicated by increased amniotic fluid interleukin (IL)-6 concentration (≥3000 pg/mL). STUDY DESIGN: A cohort of 114 women with singleton pregnancies complicated by PPROM between 24+0 and 36+6 weeks of gestation were included. Amniotic fluid samples were obtained via amniocentesis upon admission. MIAC diagnosis involved aerobic and anaerobic cultures, as well as polymerase chain reaction (PCR) analysis of the amniotic fluid. Immunoassay tests and enzyme-linked immunosorbent assay (ELISA) were used to determine IL-6 and IP-10 concentrations, respectively. RESULTS: Among the participants, 19.3 % and 15.8 % had MIAC and IAI, respectively. The occurrence of intra-amniotic inflammatory changes associated with chronic inflammation in the placenta was similar between women with and without MIAC (25 % vs. 40.9 %, p = 0.136, adjusted p = 0.213). The rate of intra-amniotic inflammatory changes associated with chronic inflammation in the placenta was significantly higher in women with IAI compared to those without, after adjusting for gestational age at sampling (55.6 % vs. 22.9 %, p = 0.005, adjusted p = 0.011). CONCLUSION: This study revealed comparable rates of intra-amniotic inflammatory changes associated with chronic inflammation in the placenta in women with and without MIAC, but a higher prevalence of intra-amniotic inflammatory changes associated with chronic inflammation in the placenta in women with IAI. These findings suggest involvement of chronic inflammation even in women with PPROM with acute intra-amniotic inflammation.
- MeSH
- chemokin CXCL10 metabolismus MeSH
- chorioamnionitida * diagnóza MeSH
- gestační stáří MeSH
- interferon gama MeSH
- lidé MeSH
- novorozenec MeSH
- placenta metabolismus MeSH
- plodová voda metabolismus MeSH
- předčasný odtok plodové vody * diagnóza MeSH
- těhotenství MeSH
- zánět komplikace MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Human induced pluripotent stem cell line was generated from commercially available primary human prostate fibroblasts HPrF derived from a fetus, aged 18-24 weeks of gestation. The fibroblast cell line was reprogrammed with Yamanaka factors (OCT4, SOX2, c-MYC, KLF4) using CytoTuneTM-iPS 2.0 Sendai Reprogramming Kit. Pluripotency of the derived transgene-free iPS cell line was confirmed both in vitro by detecting the expression of factors of pluripotency on a single-cell level, and in vivo using teratoma formation assay. This iPS cell line will be a useful tool for studying both normal prostate development and prostate cancer disease.
- MeSH
- fibroblasty * cytologie metabolismus MeSH
- indukované pluripotentní kmenové buňky * cytologie metabolismus MeSH
- lidé MeSH
- plod * cytologie embryologie MeSH
- přeprogramování buněk MeSH
- prostata * cytologie embryologie MeSH
- techniky buněčného přeprogramování * MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A human induced pluripotent stem cell line was generated from cancer-associated fibroblasts of a 68-years old patient with diagnosed prostate adenocarcinoma (PCa). The fibroblast cell line was reprogrammed with Epi5TM Episomal iPSC Reprogramming Kit. Pluripotency of the derived transgene-free iPS cell line was confirmed both in vitro by detecting expression of factors of pluripotency on a single-cell level, and also in vivo using teratoma formation assay. This new iPS cell line may be used for differentiation into different prostate-specific cell types in differentiation studies.
- MeSH
- fibroblasty asociované s nádorem metabolismus MeSH
- fibroblasty metabolismus MeSH
- indukované pluripotentní kmenové buňky metabolismus MeSH
- lidé MeSH
- nádory prostaty genetika MeSH
- senioři MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
A new microsporidium is reported from the small spruce bark beetle, Ips amitinus: Microsporidium sp. with uninucleate oval spores measuring 3.5 × 2.5 μm; infecting cells of the midgut epithelium, midgut muscles, the fat body, the Malpighian tubules, and the gonads of adult beetles collected in Austria. Seven other pathogens were found in beetles collected from Austria, the Czech Republic, and Finland. Six of them were already known from I. amitinus. Nosema cf. typographi is recorded for the first time in the overwintering generation of I. amitinus from the Czech Republic.
- MeSH
- mikrosporidia neklasifikovaná klasifikace genetika izolace a purifikace MeSH
- nosatcovití mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Finsko MeSH
- Rakousko MeSH
Induced pluripotent stem (iPS) cells are derived from differentiated cells by different reprogramming techniques, by introducing specific transcription factors responsible for pluripotency. Induced pluripotent stem cells can serve as an excellent source for differentiated neural stem/progenitor cells (NSCs/NPs). Several methods and protocols are utilized to create a robust number of NSCs/NPs without jeopardizing the safety issues required for in vivo applications. A variety of disease-specific iPS cells have been used to study nervous system diseases. In this chapter, we will focus on some of the derivation and differentiation approaches and the application of iPS-NPs in the treatment of spinal cord injury and stroke.
- MeSH
- buněčná diferenciace * MeSH
- cévní mozková příhoda patologie terapie MeSH
- indukované pluripotentní kmenové buňky cytologie MeSH
- lidé MeSH
- modely neurologické * MeSH
- nervové kmenové buňky cytologie MeSH
- poranění míchy patologie terapie MeSH
- přeprogramování buněk MeSH
- transkripční faktory metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH