The 3-mercaptopyruvate sulfurtransferase (MPST) is a protein persulfidase, occurring mainly in mitochondria. Although function of this protein in cancer cells has been already studied, no clear outcome can be postulated up to now. Therefore, we focused on the determination of function of MPST in colon (HCT116 cells)/colorectal (DLD1 cells) cancers. In silico analysis revealed that in gastrointestinal cancers, MPST together with its binding partners can be either of a high risk or might have a protective effect. Silencing of MPST gene resulted in decreased ATP, while acetyl-CoA levels were elevated. Increased apoptosis was detected in cells with silenced MPST gene, which was accompanied by decrease in mitochondrial membrane potential, but no changes in IP3 receptor's protein. Mitochondria underwent activation of fission and elevated DRP1 expression after MPST silencing. Proliferation and migration of DLD1 and HCT116 cells were markedly affected, showing the importance of MPST protein in colon/colorectal cancer development.
- MeSH
- apoptóza MeSH
- dynaminy metabolismus genetika MeSH
- HCT116 buňky MeSH
- kolorektální nádory * metabolismus patologie enzymologie genetika MeSH
- lidé MeSH
- membránový potenciál mitochondrií MeSH
- mitochondrie metabolismus MeSH
- nádorové buněčné linie MeSH
- nádory tračníku metabolismus patologie genetika enzymologie MeSH
- pohyb buněk MeSH
- proliferace buněk MeSH
- sulfurtransferasy * metabolismus genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Ťažká kombinovaná imunodeficiencia (Severe combined immunodeficiency – SCID) je život ohrozujúce ochorenie, ktoré je charakterizované poruchou bunkovej a humorálnej imunity. Je jednou z najzávažnejších ochorení v skupine primárnych imunodeficiencií. V dôsledku závažnej infekcie môže viesť k úmrtiu už v skorom dojčenskom období. Cieľ štúdie: Cieľom práce bolo analyzovanie jednotlivých charakteristík v skupine pacientov so SCID na Slovensku a vyhodnotenie výsledkov liečby transplantáciou krvotvorných buniek za účelom zmapovania celkového stavu diagnostiky, terapie a prognózy pacientov s diagnózou SCID. Metódy: Retrospektívne bol hodnotený súbor pacientov s ťažkou kombinovanou imunodeficienciou. Za obdobie 15 rokov (2003–2017) bolo celkovo liečených v Národnom ústave detských chorôb (NÚDCH) 16 detí s diagnózou SCID. Z toho 11 podstúpilo alogénnu transplantáciu krvotvorných buniek (TKB) na Transplantačnej jednotke kostnej drene NÚDCH v Bratislave. Výsledky: Pravdepodobnosť 2-ročného celkového prežívania bola v celom súbore pacientov SCID 68,8 %. V súbore 16 detí bol genetický defekt potvrdený u 13 pacientov s najčastejšou mutáciou v géne IL2RG. Medián veku stanovania diagnózy SCID bol 122 dní, s čím je asociovaný aj vyšší výskyt infekčných komplikácií. U 73 % pacientov bola potvrdená infekcia pred TKB. V súbore sme zaznamenali 4 úmrtia v dôsledku infekcie pred TKB. Analýza suchej kvapky krvi, získaná eluáciou skríningovej karty, potvrdila retrospektívne nízke hodnoty TREC u 100 % pacientov. Pravdepodobnosť 2-ročného celkového prežívania v skupine pacientov SCID po TKB bola 90,9 %. Záver: Včasné stanovenie diagnózy SCID, pred objavením sa infekčných komplikácií, zvyšuje šance pripraviť pacienta bezpečne k transplantácii krvotvorných buniek. Diagnostika TREC v našom súbore potvrdila výbornú senzitivitu a špecificitu metodiky a jej implementácia do novorodeneckého skríningu podľa vzoru mnohých krajín vo svete by bola prínosom pre zlepšenie morbidity a mortality pacientov s diagnózou SCID. Do jej zavedenia je nevyhnutné zefektívnenie skorej diagnostiky na základe charakteristických klinických príznakov a následné centralizovanie pacientov do centier so skúsenosťami s liečbou SCID pacientov.
Severe combined immunodeficiency (SCID) is a life-threatening condition characterized by impaired cellular and humoral immune responses. SCID represents one of the most severe forms of primary immunodeficiency (PID) disorders. As a result of severe infection can lead to early infant death. Objective: The aim of the study was to analyze the individual characteristics in the group of patients with SCID in Slovakia and to evaluate the results of treatment with hematopoietic stem cell transplantation in order to map the overall state of diagnosis, therapy and prognosis of patients diagnosed with SCID. Methods: In clinical part we have retrospectively evaluated the group of pediatric patients with severe combined immunodeficiency. In the period of 15 years (2003–2017), a total of 16 patients with SCID were treated in National Institute of Children´s Diseases. Of the 16 patients 11 underwent hematopoetic stem cell transplantation. Results: 2-year overall survival rate of children with SCID was 68.8%. Genetic defect was confirmed in 13 children with IL2RG as the most common mutation. The median age at the diagnosis was 122 days which was associated with higher incidence of infection before HSCT. Infection before HSCT was confirmed in 73% of patients. Due to infectious complications four patients died before HSCT. 100% patients had absence of TREC episomes which was eluated by analysis of dry blood spots. 2-year overall survival rate of children after HSCT was 90.9%. Conclusion: Early diagnosis of SCID, before the onset of severe infections, increases the chances of getting the patients safely to HSCT. The TREC diagnosis in our sample confirmed the excellent sensitivity and specificity of the method and its implementation in neonatal screening, following the example of many countries around the world, would be beneficial in improving the morbidity and mortality of patients diagnosed with SCID. Until its introduction it is necessary to streamline early diagnosis based on characteristic clinical symptoms and the subsequent centralization of patients into centers with experience in the treatment of SCID patients.
The presence of SARS-CoV-2 RNA in wastewater was first reported in March 2020. Over the subsequent months, the potential for wastewater surveillance to contribute to COVID-19 mitigation programmes has been the focus of intense national and international research activities, gaining the attention of policy makers and the public. As a new application of an established methodology, focused collaboration between public health practitioners and wastewater researchers is essential to developing a common understanding on how, when and where the outputs of this non-invasive community-level approach can deliver actionable outcomes for public health authorities. Within this context, the NORMAN SCORE "SARS-CoV-2 in sewage" database provides a platform for rapid, open access data sharing, validated by the uploading of 276 data sets from nine countries to-date. Through offering direct access to underpinning meta-data sets (and describing its use in data interpretation), the NORMAN SCORE database is a resource for the development of recommendations on minimum data requirements for wastewater pathogen surveillance. It is also a tool to engage public health practitioners in discussions on use of the approach, providing an opportunity to build mutual understanding of the demand and supply for data and facilitate the translation of this promising research application into public health practice.
- MeSH
- COVID-19 * MeSH
- lidé MeSH
- odpadní voda MeSH
- RNA virová MeSH
- SARS-CoV-2 * MeSH
- veřejné zdravotnictví MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Publikační typ
- abstrakt z konference MeSH
BACKGROUND: The Roma are a European ethnic minority threatened by several recessive diseases. Variants in MANBA cause a rare lysosomal storage disorder named beta-mannosidosis whose clinical manifestation includes deafness and mental retardation. Since 1986, only 23 patients with beta-mannosidosis and biallelic MANBA variants have been described worldwide. RESULTS: We now report on further 10 beta-mannosidosis patients of Roma origin from eight families in the Czech and Slovak Republics with hearing loss, mental retardation and homozygous pathogenic variants in MANBA. MANBA variant c.2158-2A>G screening among 345 anonymized normal hearing controls from Roma populations revealed a carrier/heterozygote frequency of 3.77%. This is about 925 times higher than the frequency of this variant in the gnomAD public database and classifies the c.2158-2A>G variant as a prevalent, ethnic-specific variant causing hearing loss and mental retardation in a homozygous state. The frequency of heterozygotes/carriers is similar to another pathogenic variant c.71G>A (p.W24*) in GJB2, regarded as the most frequent variant causing deafness in Roma populations. CONLCUSION: Beta-mannosidosis, due to a homozygous c.2158-2A>G MANBA variant, is an important and previously unknown cause of hearing loss and mental retardation among Central European Roma.
- MeSH
- beta-mannosidóza * MeSH
- etnicita MeSH
- hluchota * genetika MeSH
- lidé MeSH
- menšiny MeSH
- nedoslýchavost * genetika MeSH
- Romové * genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Slovenská republika MeSH
- Publikační typ
- abstrakt z konference MeSH
BACKGROUND: Knowledge about the expression and thus a role of enzymes that produce endogenous H2S - cystathionine-β-synthase, cystathionine γ-lyase and mercaptopyruvate sulfurtransferase - in renal tumors is still controversial. In this study we aimed to determine the expression of these enzymes relatively to the expression in unaffected part of kidney from the same patient and to found relation of these changes to apoptosis. To evaluate patient's samples, microarray and immunohistochemistry was used. METHODS: To determine the physiological importance, we used RCC4 stable cell line derived from clear cell renal cell carcinoma, where apoptosis induction by a mixture of five chemotherapeutics with/without silencing of H2S-producing enzymes was detected. Immunofluorescence was used to determine each enzyme in the cells. RESULTS: In clear cell renal cell carcinomas, expression of H2S-producing enzymes was mostly decreased compared to a part of kidney that was distal from the tumor. To evaluate a potential role of H2S-producing enzymes in the apoptosis induction, we used RCC4 stable cell line. We have found that silencing of cystathionine-β-synthase and cystathionine γ-lyase prevented induction of apoptosis. Immunofluorescence staining clearly showed that these enzymes were upregulated during apoptosis in RCC4 cells. CONCLUSION: Based on these results we concluded that in clear cell renal cell carcinoma, reduced expression of the H2S-producing enzymes, mainly cystathionine γ-lyase, might contribute to a resistance to the induction of apoptosis. Increased production of the endogenous H2S, or donation from the external sources might be of a therapeutic importance in these tumors.
- MeSH
- apoptóza * MeSH
- cystathionin-beta-synthasa genetika metabolismus MeSH
- cystathionin-gama-lyasa genetika metabolismus MeSH
- dospělí MeSH
- karcinom z renálních buněk patologie chirurgie MeSH
- ledviny metabolismus patologie chirurgie MeSH
- lidé středního věku MeSH
- lidé MeSH
- malá interferující RNA metabolismus MeSH
- nádorové buněčné linie MeSH
- nádory ledvin patologie chirurgie MeSH
- nefrektomie MeSH
- RNA interference MeSH
- senioři MeSH
- sulfan metabolismus MeSH
- upregulace MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: In the present study we aimed: 1) To establish the prevalence and clinical impact of DFNB49 mutations in deaf Roma from 2 Central European countries (Slovakia and Hungary), and 2) to analyze a possible common origin of the c.1331+2T>C mutation among Roma and Pakistani mutation carriers identified in the present and previous studies. METHODS: We sequenced 6 exons of the MARVELD2 gene in a group of 143 unrelated hearing impaired Slovak Roma patients. Simultaneously, we used RFLP to detect the c.1331+2T>C mutation in 85 Hungarian deaf Roma patients, control groups of 702 normal hearing Romanies from both countries and 375 hearing impaired Slovak Caucasians. We analyzed the haplotype using 21 SNPs spanning a 5.34Mb around the mutation c.1331+2T>C. RESULTS: One pathogenic mutation (c.1331+2T>C) was identified in 12 homozygous hearing impaired Roma patients. Allele frequency of this mutation was higher in Hungarian (10%) than in Slovak (3.85%) Roma patients. The identified common haplotype in Roma patients was defined by 18 SNP markers (3.89 Mb). Fourteen common SNPs were also shared among Pakistani and Roma homozygotes. Biallelic mutation carriers suffered from prelingual bilateral moderate to profound sensorineural hearing loss. CONCLUSIONS: We demonstrate different frequencies of the c.1331+2T>C mutation in hearing impaired Romanies from 3 Central European countries. In addition, our results provide support for the hypothesis of a possible common ancestor of the Slovak, Hungarian and Czech Roma as well as Pakistani deaf patients. Testing for the c.1331+2T>C mutation may be recommended in GJB2 negative Roma cases with early-onset sensorineural hearing loss.
- MeSH
- alely MeSH
- efekt zakladatele MeSH
- exony genetika MeSH
- frekvence genu MeSH
- genotyp MeSH
- haplotypy genetika MeSH
- jednonukleotidový polymorfismus * MeSH
- kojenec MeSH
- lidé MeSH
- MARVELD2 protein genetika MeSH
- mutace * MeSH
- nedoslýchavost vrozené etnologie genetika MeSH
- prevalence MeSH
- Romové genetika MeSH
- sekvenční homologie nukleových kyselin MeSH
- věk při počátku nemoci MeSH
- Check Tag
- kojenec MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- srovnávací studie MeSH
- Geografické názvy
- Česká republika MeSH
- Maďarsko MeSH
- Pákistán MeSH
- Slovenská republika MeSH
OBJECTIVE: Individuals with decreased thiopurine methyltransferase (TPMT) activity are at risk of adverse effects of thiopurine administration whereas its increased activity may inactivate drugs faster. We evaluated genotype-phenotype correlations in patients with suspected hematological malignancies and inflammatory bowel disease from our region based on findings of nonlinear TPMT enzyme kinetics previously unreported. PATIENTS AND METHODS: The study group comprised 267 individuals. They were screened for the most common variants of low TPMT activity. TPMT activity was measured in erythrocytes using the HPLC rate-blanked method. RESULTS: Thirty-three patients (12.4%) were heterozygous (26 were TPMT*1/*3A, 5 TPMT*1/*2, 2 TPMT *1/*3C) and 1 was a compound heterozygote (*2/*3A). Normal and low normal TPMT activities substantially overlapped in wild-type and heterozygous individuals, whereas high activities were found in 29 wild-type genotyped patients. Extreme and life-threatening toxicity was observed in the compound heterozygote patient. CONCLUSION: Activity measurement performed at diagnosis provides clinicians with information on immediate pharmacokinetic-related adverse events and/or hypermetabolism, and genotyping may indicate the rate of pharmacodynamic thioguanine nucleotide accumulation due to slower overall thiopurine metabolism.
- MeSH
- asparaginasa terapeutické užití MeSH
- cyklofosfamid terapeutické užití MeSH
- cytarabin terapeutické užití MeSH
- daunomycin terapeutické užití MeSH
- erytrocytární membrána enzymologie MeSH
- genetické asociační studie MeSH
- idiopatické střevní záněty krev MeSH
- lidé MeSH
- lymfoblastická leukemie-lymfom z prekurzorových T-buněk farmakoterapie genetika MeSH
- merkaptopurin terapeutické užití MeSH
- methotrexát terapeutické užití MeSH
- methyltransferasy nedostatek genetika metabolismus MeSH
- mladiství MeSH
- polymerázová řetězová reakce MeSH
- prednison terapeutické užití MeSH
- protokoly protinádorové kombinované chemoterapie terapeutické užití MeSH
- vinkristin terapeutické užití MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Check Tag
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Slovenská republika MeSH
