Dyskeratosis congenita (DC) is a rare inherited bone marrow failure syndrome, caused by genetic mutations that principally affect telomere biology. Approximately 35% of cases remain uncharacterised at the genetic level. To explore the genetic landscape, we conducted genetic studies on a large collection of clinically diagnosed cases of DC as well as cases exhibiting features resembling DC, referred to as 'DC-like' (DCL). This led us to identify several novel pathogenic variants within known genetic loci and in the novel X-linked gene, POLA1. In addition, we have also identified several novel variants in POT1 and ZCCHC8 in multiple cases from different families expanding the allelic series of DC and DCL phenotypes. Functional characterisation of novel POLA1 and POT1 variants, revealed pathogenic effects on protein-protein interactions with primase, CTC1-STN1-TEN1 (CST) and shelterin subunit complexes, that are critical for telomere maintenance. ZCCHC8 variants demonstrated ZCCHC8 deficiency and signs of pervasive transcription, triggering inflammation in patients' blood. In conclusion, our studies expand the current genetic architecture and broaden our understanding of disease mechanisms underlying DC and DCL disorders.
- MeSH
- dítě MeSH
- dyskeratosis congenita * genetika MeSH
- lidé MeSH
- mutace MeSH
- proteiny vázající telomery * genetika metabolismus MeSH
- shelterinový komplex MeSH
- telomery * genetika metabolismus MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Protein phosphatase magnesium-dependent 1 delta (PPM1D) terminates the cell cycle checkpoint by dephosphorylating the tumour suppressor protein p53. By targeting additional substrates at chromatin, PPM1D contributes to the control of DNA damage response and DNA repair. Using proximity biotinylation followed by proteomic analysis, we identified a novel interaction between PPM1D and the shelterin complex that protects telomeric DNA. In addition, confocal microscopy revealed that endogenous PPM1D localises at telomeres. Further, we found that ATR phosphorylated TRF2 at S410 after induction of DNA double strand breaks at telomeres and this modification increased after inhibition or loss of PPM1D. TRF2 phosphorylation stimulated its interaction with TIN2 both in vitro and at telomeres. Conversely, induced expression of PPM1D impaired localisation of TIN2 and TPP1 at telomeres. Finally, recruitment of the DNA repair factor 53BP1 to the telomeric breaks was strongly reduced after inhibition of PPM1D and was rescued by the expression of TRF2-S410A mutant. Our results suggest that TRF2 phosphorylation promotes the association of TIN2 within the shelterin complex and regulates DNA repair at telomeres.
- MeSH
- ektodermální dysplazie 1 anhidrotická diagnóza genetika terapie MeSH
- lidé MeSH
- mezioborová komunikace MeSH
- mladý dospělý MeSH
- Check Tag
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Úvod: V současnosti není k dispozici vhodná strategie k léčbě vulvodynie u žen. Extrakorporeální rázová vlna (Extracorporeal shock wave therapy – ESWT) je rozšířena pro léčbu muskuloskeletárních poruch, svalové spasticity, hypertonu, renální a biliární lithiázy, urologických a v poslední době i andrologických onemocnění. Rozhodli jsme se vyšetřit užití ESWT u vulvodynie u žen. Metody: Prospektivní studie byla prováděna v letech 2015 – 2018. Studie proveditelnosti zahrnovala 30 žen s vulvodynií nejméně 3 měsíce. Pacientky byly léčeny perineálně aplikovanou ESWT 1x týdně (3 000 pulzů vždy po 4 následující týdny). Použité zařízení bylo standardní elektromagnetický unit s fokusovanou rázovou hlavicí. Ošetřovaná oblast byla měněna vždy po 500 pulzech. Šesti okrsky byla ošetřena celá vulva a perineum. Stupeň bolesti byl hodnocen pomocí vizuální analogové škály o 11 stupních (VAS, 0 – 10) před a po léčbě. Cotton Swab test (CST, Goetsch škála 0 – 4) ve stejných termínech. Follow-up byla provedena po 1, 4 a 12 týdnech po ukončení poslední aplikace ESWT. Souhlas Etické komise a písemné informované souhlasy pacientek byly zajištěny. Výsledky: 29 žen dokončilo studii. Ve VAS a CST testování jsme prokázali signifikantní rozdíly mezi výsledky před aplikací a všemi výsledky follow-up (P < 0,001; P < 0,05) v tomto pořadí. Redukce bolesti ve VAS byla 53 % a CST 47 % při posledním testování Závěry: ESWT se zdá významně redukovat vnímání bolesti v naší skupině léčených žen. Výsledky nás povzbuzují k dalšímu zkoumání této moderní techniky. Tato metoda je jednoduše opakovatelná, levná a bez prokazatelných vedlejších účinků.
Introduction: There are no sufficient therapy strategies for vulvodynia in women. Extracorporeal shock wave therapy (ESWT) is widely used for the treatment of musculoskeletal diseases, muscle spasticity, hypertonus, renal and biliary calculi, urological and recently andrological disorders. We examined the effect of ESWT on vulvodynia in women. Methods: A prospective study was conducted between 2015 and 2018 years. The feasibility study involved thirty women with vulvodynia for at least 3 months. The patients received perineally applied ESWT treatment weekly (3 000 pulses each for 4 consecutive weeks). The device used was a standard electromagnetic shock wave unit with a focused shock wave handpiece. The access area was changed after every 500 pulses. Six areas covered the whole vulva and perineum.The grade of pain was evaluated with the visual analogue scale of 11 degrees (VAS, 0-10) before and after the treatment and Cotton Swab test (CST, Goetsch scale 0-4) at the same terms. Follow-ups were done after 1, 4, and 12 weeks having finished the last ESWT. Ethical committee agreement and written consents were provided. Results: 29 women completed the study. In VAS and CST testing we proved significant differences between before and all the follow-ups (P < 0.001; P < 0.05) respectively. The reduction of pain in VAS was 53 %; CST 47 % in the last testing. Conclusions: ESWT seems to reduce significantly the pain perception in our group of treated patients. We are encouraged to explore this technique further. The method is easily repeatable, inexpensive, and free of any side effects.
OBJECTIVE: To analyze the cervical microbiota in women with preterm prelabor rupture of membranes (PPROM) by pyrosequencing and to document associations between cervical microbiota, cervical inflammatory response, microbial invasion of the amniotic cavity (MIAC), histological chorioamnionitis, and intraamniotic infection (IAI). STUDY DESIGN: Sixty-one women with singleton pregnancies complicated by PPROM were included in the study. Specimens of cervical and amniotic fluid were collected on admission. The cervical microbiota was assessed by 16S rRNA gene sequencing by pyrosequencing. Interleukin (IL)-6 concentration in the cervical fluid and amniotic fluid was measured by ELISA and lateral flow immunoassay, respectively. RESULTS: Four bacterial community state types [CST I (Lactobacillus crispatus dominated), CST III (Lactobacillus iners dominated), CST IV-A (non-Lactobacillus bacteria dominated), and CST IV-B (Gardnerella vaginalis and Sneathia sanguinegens dominated)] were observed in the cervical microbiota of women with PPROM. Cervical fluid IL-6 concentrations differed between CSTs (CST I = 145 pg/mL, CST III = 166 pg/mL, CST IV-A = 420 pg/mL, and CST IV-B = 322 pg/mL; p = 0.004). There were also differences in the rates of MIAC, of both MIAC and histological chorioamnionitis, and of IAI among CSTs. No difference in the rate of histological chorioamnionitis was found among CSTs. CONCLUSIONS: The cervical microbiota in PPROM women in this study was characterized by four CSTs. The presence of non-Lactobacillus CSTs was associated with a strong cervical inflammatory response and higher rates of MIAC, both MIAC and histological chorioamnionitis, and IAI representing a PPROM subtype with pronounced inflammation. CST I represents the dominant type of PPROM with a low rate of MIAC, IAI, and the combination of MIAC and histological chorioamnionitis.
- MeSH
- amnion mikrobiologie MeSH
- cervix uteri mikrobiologie patologie MeSH
- chorioamnionitida mikrobiologie MeSH
- demografie MeSH
- dospělí MeSH
- druhová specificita MeSH
- interleukin-6 metabolismus MeSH
- lidé MeSH
- mikrobiota * MeSH
- mladý dospělý MeSH
- novorozenec MeSH
- plodová voda metabolismus MeSH
- předčasná porodní činnost mikrobiologie MeSH
- předčasný odtok plodové vody mikrobiologie MeSH
- těhotenství MeSH
- tělesné tekutiny mikrobiologie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- novorozenec MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
