nDNA
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INTRODUCTION: We evaluated the levels of cell-free nuclear DNA (nDNA) and cell-free mitochondrial DNA (mtDNA) in the amniotic fluid supernatant from pregnancies complicated by preterm prelabor rupture of membranes (PPROM) based on evidence of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI). MATERIAL AND METHODS: A total of 155 women with PPROM were included in this study. Amniotic fluid samples were obtained by transabdominal amniocentesis. The levels of cell-free nDNA and mtDNA in the amniotic fluid supernatant were assessed and quantified by real-time polymerase chain reaction. RESULTS: The levels of cell-free nDNA and mtDNA were higher in women with MIAC and IAI than in women without these conditions (nDNA: with MIAC: median 3.9 × 104 genome equivalent [GE]/mL vs without MIAC: median 1.2 × 104 GE/mL, with IAI: median: 5.3 × 104 GE/mL vs without IAI: median 1.2 × 104 GE/mL; mtDNA: with MIAC: median 9.2 × 105 GE/mL vs without MIAC: median 2.5 × 105 GE/mL, with IAI: median 1.1 × 106 GE/mL vs without IAI: median 2.5 × 105 ; all P values ≤ 0.01). Women with the microbial-associated IAI showed the highest levels of cell-free nDNA and mtDNA. CONCLUSIONS: Cell-free nDNA and mtDNA are constituents of the amniotic fluid supernatant from PPROM pregnancies. Both cell-free nDNA and mtDNA are involved in the intra-amniotic inflammatory response in women with PPROM.
- MeSH
- amniocentéza MeSH
- bakteriální infekce metabolismus MeSH
- Chlamydia trachomatis MeSH
- chorioamnionitida metabolismus MeSH
- dospělí MeSH
- gestační stáří MeSH
- interleukin-6 metabolismus MeSH
- kohortové studie MeSH
- kultivační techniky MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- mitochondriální DNA metabolismus MeSH
- Mycoplasma hominis MeSH
- plodová voda metabolismus mikrobiologie MeSH
- polymerázová řetězová reakce MeSH
- předčasný odtok plodové vody metabolismus MeSH
- retrospektivní studie MeSH
- RNA ribozomální 16S analýza MeSH
- těhotenství MeSH
- Ureaplasma MeSH
- volné cirkulující nukleové kyseliny metabolismus MeSH
- zánět metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Mitochondria are vital organelles with their own DNA (mtDNA). mtDNA is circular and composed of heavy and light chains that are structurally more accessible than nuclear DNA (nDNA). While nDNA is typically diploid, the number of mtDNA copies per cell is higher and varies considerably during development and between tissues. Compared with nDNA, mtDNA is more prone to damage that is positively linked to many diseases, including cancer. Similar to nDNA, mtDNA undergoes repair processes, although these mechanisms are less well understood. In this review, we discuss the various forms of mtDNA damage and repair and their association with cancer initiation and progression. We also propose horizontal mitochondrial transfer as a novel mechanism for replacing damaged mtDNA.
- MeSH
- lidé MeSH
- mitochondriální DNA * genetika MeSH
- mitochondrie * genetika metabolismus MeSH
- nádory * genetika patologie MeSH
- oprava DNA * MeSH
- poškození DNA * genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Huntington's disease (HD) is a progressive neurodegenerative disorder primarily affecting the basal ganglia and is caused by expanded CAG repeats in the huntingtin gene. Except for CAG sizing, mitochondrial and nuclear DNA (mtDNA and nDNA) parameters have not yet proven to be representative biomarkers for disease and future therapy. Here, we identified a general suppression of genes associated with aerobic metabolism in peripheral blood mononuclear cells (PBMCs) from HD patients compared to controls. In HD, the complex II subunit SDHB was lowered although not sufficiently to affect complex II activity. Nevertheless, we found decreased level of factors associated with mitochondrial biogenesis and an associated dampening of the mitochondrial DNA damage frequency in HD, implying an early defect in mitochondrial activity. In contrast to mtDNA, nDNA from HD patients was four-fold more modified than controls and demonstrated that nDNA integrity is severely reduced in HD. Interestingly, the level of nDNA damage correlated inversely with the total functional capacity (TFC) score; an established functional score of HD. Our data show that PBMCs are a promising source to monitor HD progression and highlights nDNA damage and diverging mitochondrial and nuclear genome responses representing early cellular impairments in HD.
- MeSH
- dospělí MeSH
- Huntingtonova nemoc genetika patologie MeSH
- leukocyty mononukleární metabolismus patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mitochondriální DNA analýza genetika MeSH
- mitochondrie metabolismus patologie MeSH
- mladý dospělý MeSH
- nestabilita genomu * MeSH
- poškození DNA * MeSH
- senioři MeSH
- studie případů a kontrol MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
PurposeDiagnosing primary mitochondrial diseases (MDs) is challenging in clinical practice. The mitochondrial disease criteria (MDC) have been developed to quantify the clinical picture and evaluate the probability of an underlying MD and the need for a muscle biopsy. In this new genetic era with next-generation sequencing in routine practice, we aim to validate the diagnostic value of MDC.MethodsWe retrospectively studied MDC in a multicenter cohort of genetically confirmed primary MD patients.ResultsWe studied 136 patients (61 male, 91 nuclear DNA (nDNA) mutations). Forty-five patients (33%) had probable MD and 69 (51%) had definite MD according to the MDC. A muscle biopsy was performed in 63 patients (47%). Patients with nDNA mutations versus mitochondrial DNA mutations were younger (6.4 ± 9.7 versus 19.5 ± 17.3 y) and had higher MDC (7.07 ± 1.12/8 versus 5.69 ± 1.94/8). At a cutoff of 6.5/8, the sensitivity to diagnose patients with nDNA mutations is 72.5% with a positive predictive value of 69.5%. In the nDNA mutation group, whole-exome sequencing could diagnose patients with lower scores (MDC (6.84 ± 1.51/8) compared to Sanger sequencing MDC (7.44 ± 1.13/8, P = 0.025)). Moreover 7/8 patients diagnosed with possible MD by MDC were diagnosed by whole-exome sequencing.ConclusionMDC remain very useful in the clinical diagnosis of MD, in interpreting whole-exome results and deciding on the need for performing muscle biopsy.
- MeSH
- dítě MeSH
- dospělí MeSH
- genetická predispozice k nemoci MeSH
- genetické asociační studie MeSH
- genetické testování metody MeSH
- genom mitochondriální * MeSH
- genomika * metody MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- mitochondriální geny MeSH
- mitochondriální nemoci diagnóza genetika MeSH
- mitochondrie genetika metabolismus MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mutace MeSH
- novorozenec MeSH
- předškolní dítě MeSH
- průběh práce MeSH
- retrospektivní studie MeSH
- senioři MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- novorozenec MeSH
- předškolní dítě MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- práce podpořená grantem MeSH
BACKGROUND: Huntington's disease (HD) is a devastating neurodegenerative disorder caused by CAG triplet expansions in the huntingtin gene. Oxidative stress is linked to HD pathology, although it is not clear whether this is an effect or a mediator of disease. The transgenic (TgHD) minipig expresses the N-terminal part of human-mutated huntingtin and represents a unique model to investigate therapeutic strategies towards HD. A more detailed characterization of this model is needed to fully utilize its potential. METHODS: In this study, we focused on the molecular and cellular features of fibroblasts isolated from TgHD minipigs and the wild-type (WT) siblings at different ages, pre-symptomatic at the age of 24-36 months and with the onset of behavioural symptoms at the age of 48 months. We measured oxidative stress, the expression of oxidative stress-related genes, proliferation capacity along with the expression of cyclin B1 and D1 proteins, cellular permeability, and the integrity of the nuclear DNA (nDNA) and mitochondrial DNA in these cells. RESULTS: TgHD fibroblasts isolated from 48-month-old animals showed increased oxidative stress, which correlated with the overexpression of SOD2 encoding mitochondrial superoxide dismutase 2, and the NEIL3 gene encoding DNA glycosylase involved in replication-associated repair of oxidized DNA. TgHD cells displayed an abnormal proliferation capacity and permeability. We further demonstrated increased nDNA damage in pre-symptomatic TgHD fibroblasts (isolated from animals aged 24-36 months). CONCLUSIONS: Our results unravel phenotypic alterations in primary fibroblasts isolated from the TgHD minipig model at the age of 48 months. Importantly, nDNA damage appears to precede these phenotypic alterations. Our results highlight the impact of fibroblasts from TgHD minipigs in studying the molecular mechanisms of HD pathophysiology that gradually occur with age.
- MeSH
- buněčné dělení MeSH
- fenotyp MeSH
- fibroblasty metabolismus MeSH
- geneticky modifikovaná zvířata MeSH
- lidé MeSH
- miniaturní prasata MeSH
- mitochondriální DNA genetika MeSH
- N-glykosylhydrolasy biosyntéza genetika MeSH
- oxidační stres MeSH
- peroxidace lipidů MeSH
- poškození DNA MeSH
- prasata MeSH
- primární buněčná kultura MeSH
- protein huntingtin genetika metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- regulace genové exprese MeSH
- stárnutí metabolismus MeSH
- superoxiddismutasa biosyntéza genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Naive use of molecular data may lead to ambiguous conclusions, especially within the context of "cryptic" species. Here, we integrated molecular and morphometric data to evaluate phylogenetic relationships in the widespread terrestrial micro-snail genus, Euconulus. We analyzed mitochondrial (16S + COII) and nuclear (ITS1 + ITS2) sequence across 94 populations from Europe, Asia and North America within the nominate species E. alderi, E. fulvus and E. polygyratus, and used the southeastern USA E. chersinus, E. dentatus, and E. trochulus as comparative outgroups. Phylogeny was reconstructed using four different reconstruction methods to identify robust, well-supported topological features. We then performed discriminant analysis on shell measurements between these genetically-identified species-level clades. These analyses provided evidence for a biologically valid North American "cryptic" species within E. alderi. However, while highly supported polyphyletic structure was also observed within E. fulvus, disagreement in placement of individuals between mtDNA and nDNA clades, lack of morphological differences, and presence of potential hybrids imply that these lineages do not rise to the threshold as biologically valid cryptic species, and rather appear to simply represent a complex of geographically structured populations within a single species. These results caution that entering into a cryptic species hypothesis should not be undertaken lightly, and should be optimally supported along multiple lines of evidence. Generally, post-hoc analyses of macro-scale features should be conducted to attempt identification of previously ignored diagnostic traits. If such traits cannot be found, i.e. in the case of potentially "fully cryptic" species, additional criteria should be met to propound a cryptic species hypothesis, including the agreement in tree topology among both mtDNA and nDNA, and little (or no) evidence of hybridization based on a critical analysis of sequence chromatograms. Even when the above conditions are satisfied, it only implies that the cryptic species hypothesis is plausible, but should optimally be subjected to further careful examination.
- MeSH
- analýza hlavních komponent MeSH
- buněčné jádro genetika MeSH
- fylogeneze MeSH
- hlemýždi klasifikace genetika MeSH
- pravděpodobnostní funkce MeSH
- respirační komplex IV klasifikace genetika MeSH
- RNA ribozomální 16S klasifikace genetika MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- sekvenční seřazení MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Amyloid-beta (Aβ) induced mitochondrial dysfunction is one of the major causes of neuronal toxicity in Alzheimer's disease. A number of recent reports suggest involvement of mitochondrial alterations through intracellular accumulation of oligomeric Aβ. These mitochondrial alterations include increased Reactive Oxygen Species (ROS), mt-DNA depletion, decreased oxidative phosphorylation and ATP production, membrane depolarization, reduced number of mitochondria etc. All these defects cumulatively caused neural toxicity and alterations in cellular energy homeostasis. On the other hand, anti-inflammatory drug aspirin is reported to promote both mitochondrial biogenesis and improvement in cellular energy status. METHODS: Taking altogether the mentioned clues, we evaluated protective effect of aspirin, if any on oligomeric Aβ42 induced toxicity and mitochondrial alterations in differentiated neuronal cells. RESULTS: A significant reduction in neuronal viability and increased apoptosis was observed in Aβ42 treated cells, as evident by MTT assay, apoptosis ELISA and immunofluorescence from β-III tubulin antibody staining of neuronal cells. A concomitant decrease was also observed in the intensity of mitotracker red FM staining and mt-DNA to nDNA ratio, suggesting mitochondrial membrane depolarization and/or reduced number of mitochondria along with depletion in mt-DNA. However, simultaneous treatment of 5 μM aspirin to oligomeric Aβ42 treated cells protected them from mitochondrial dysfunction and neurotoxicity. CONCLUSION: We suggest mitochondrial biogenesis, changes in mitochondrial membrane potential and / or inhibition of Aβ42 aggregation by aspirin as possible underlying mechanism(s).
- MeSH
- amyloidní beta-protein toxicita MeSH
- analýza rozptylu MeSH
- apoptóza účinky léků MeSH
- Aspirin farmakologie MeSH
- buněčná diferenciace účinky léků MeSH
- karcinom patologie MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- mitochondriální DNA genetika metabolismus MeSH
- mitochondrie účinky léků patologie MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádorové kmenové buňky účinky léků MeSH
- neurony účinky léků ultrastruktura MeSH
- neuroprotektivní látky farmakologie MeSH
- peptidové fragmenty toxicita MeSH
- tubulin metabolismus MeSH
- variabilita počtu kopií segmentů DNA účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Problematika epidemiologie DN a nediabetických nefropatií /NDN/ u pacientů s diabetes mellitus patří stále mezi diskutovaná témata. V práci jsme analyzovali skupinu diabetiků, u kterých byla provedena renální biopsie /RB/ v sedmi po sobě následujících letech (od 1999 též s diagnózou NDN). Sledované parametry byly následující:c sérový kreatinin /S-cr/, přítomnost hematurie /HU/, proteinurie/24-h /PU/, arteriální hypertenze /AH/, histologická diagnóza (dle SZO) a komplikace po RB. Ve sledovaném období bylo v ČR (10,3 mil. obyvatel) provedeno celkem 3878 RB. Z toho sekundární GN byly zjištěny v 993 případech (25,6 %) a u 102 RB (2,63 %) byla diagnostikována diabetická glomeruloskleróza. Mezi pacienty převládali muži (64 %), průměrný věk byl 55,6 roků (rozsah 18-80). AH byla zaznamenána v 79,8 %, mikro-HU v 58,6 %, PU nízkého stupně (<2,5 g/24-h) v 17 %, nefrotická PU (?3,0g/24-h) v 80 % a průměrná PU činila 8,9 g/24-h. Průměrná hodnota S-cr činila 244 μmol/l (rozsah 54-1002). Normální renální funkce (S-cr<110) byla zjištěna ve 22 % případů, renální insuficience /RI/ se S-cr 111-200 ve 34 %, se S-cr 201-300 ve 22 %, se S-cr 301-400 a S-cr >400 po 11 %. Roční incidence DN byla 1,41 případu/milion obyvatel. Z komplikací po RB byl ve 2 % přítomen klinicky symptomatický subkapsulární hematom. V letech 1999-2000 bylo bioptováno dalších 78 diabetiků, reprezentativní vzorek však byl získán je u 63 s diagnózou NDN. Nejčastěji byla zjištěna IgAN (17,5 %), hypertenzivní nefroskleróza a membranózní nefropatie (obě po 11,1 %) a ANCA+ vaskulitida (9,5 %). Nefrotická PU byla příto-mna u 68,1 %, mikro-HU u 59,7 %, AH u 74 % a RI u 67,9 % s průměrnou hodnotou S-cr 207 μmol/l. Závěr: DN představuje malou, ale klinicky významnou skupinu pacientů indikovaných k RB. Domníváme se, že skutečný výskyt DN a především NDN je podceněn. U pacientů s DN bylo dle očekávání zachyceno vysoké procento nefrotické PU a AH, ale byl zaznamenán též vysoký výskyt mikro-HU. Od r. 1999 cca 50 % bioptovaných diabetiků mělo NDN, nejčastěj-ší byla IgAN a překvapivě vysoké procento mělo renální vaskulitidu.
- MeSH
- biopsie statistika a číselné údaje využití MeSH
- diabetické nefropatie dějiny diagnóza MeSH
- finanční podpora výzkumu jako téma MeSH
- IgA nefropatie diagnóza MeSH
- lidé MeSH
- registrace statistika a číselné údaje MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- kongresy MeSH
- Geografické názvy
- Česká republika MeSH
We examined the global phylogeography of gypsy moth (Lymantria dispar L.) using molecular data based on mitochondrial and nuclear genes. Populations from all biogeographic regions of the native and introduced range of L. dispar, were sampled to fully document intraspecific and subspecies variation, identify potential cryptic species, and to clarify the relationships among major phylogeographic lineages. We recovered three major mtDNA lineages of L. dispar: Transcaucasia; East Asia + Japan; and Europe + Central Asia. The circumscription of these lineages is only partially consistent with the current taxonomic concept (i.e., L. dispar dispar; L. dispar asiatica; L. dispar japonica), with the following important discrepancies: (1) north-central Asian populations, including topotypical populations of L. dispar asiatica, may be more closely related to European rather than Asian segregates, which would require the synonymization of the taxon asiatica and establishment of a new name; (2) the Japanese populations (L. d. japonica) are not distinct from east Asian populations; (3) the presence of a distinct, unnamed mitogenomic lineage endemic to the Trancaucasus region. We demonstrated that the population from Transcaucasia contains the highest mitochondrial haplotype diversity among L. dispar, potentially indicative of an ancestral area for the entire dispar-group. Our study corroborates the endemic Hokkaido, Japan taxon Lymantria umbrosa (Butler) as the sister group to all other L. dispar populations, but the applicability of the names umbrosa versus hokkaidoensis Goldschmidt needs to be re-evaluated. The ancestral area analysis suggest that Japan was likely colonized via Sakhalin ∼1 Mya, in contrast to previous studies which have suggested colonization of the Japanese archipelago via the Korean Peninsula. Lastly, mitogenomic variation within L. dispar is incongruent with phylogenies based on nuclear DNA, as nDNA gene phylogenies did not recover the three major mtDNA lineages, and also failed to recover L. dispar and L. umbrosa as reciprocally monophyletic.
- MeSH
- Bayesova věta MeSH
- biologická evoluce * MeSH
- buněčné jádro genetika MeSH
- fylogeneze MeSH
- fylogeografie * MeSH
- genom mitochondriální MeSH
- haplotypy genetika MeSH
- mitochondriální DNA genetika MeSH
- můry klasifikace genetika MeSH
- pravděpodobnostní funkce MeSH
- taxonomické DNA čárové kódování MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
... , XINYI MA, LIYING YAN, JIE QIAO -- Introduction 155 -- Oogenesis: An Overview 156 -- Nuclear DNA (nDNA ...
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