OBJECTIVES: The analysis of organic acids in urine is an important part of the diagnosis of inherited metabolic disorders (IMDs), for which gas chromatography coupled with mass spectrometry is still predominantly used. METHODS: Ultra-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for urinary organic acids, acylcarnitines and acylglycines was developed and validated. Sample preparation consists only of dilution and the addition of internal standards. Raw data processing is quick and easy using selective scheduled multiple reaction monitoring mode. A robust standardised value calculation as a data transformation together with advanced automatic visualisation tools are applied for easy evaluation of complex data. RESULTS: The developed method covers 146 biomarkers consisting of organic acids (n=99), acylglycines (n=15) and acylcarnitines (n=32) including all clinically important isomeric compounds present. Linearity with r2>0.98 for 118 analytes, inter-day accuracy between 80 and 120 % and imprecision under 15 % for 120 analytes were achieved. Over 2 years, more than 800 urine samples from children tested for IMDs were analysed. The workflow was evaluated on 93 patient samples and ERNDIM External Quality Assurance samples involving a total of 34 different IMDs. CONCLUSIONS: The established LC-MS/MS workflow offers a comprehensive analysis of a wide range of organic acids, acylcarnitines and acylglycines in urine to perform effective, rapid and sensitive semi-automated diagnosis of more than 80 IMDs.

• MeSH
• chromatografie kapalinová metody   MeSH
• dítě MeSH
• lidé MeSH
• metabolické nemoci * MeSH
• organické látky MeSH
• průběh práce MeSH
• tandemová hmotnostní spektrometrie * metody   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVES: Presymptomatic detection of patients with rare diseases (RD), defined by a population frequency less than 1 : 2,000, is the task of newborn screening (NBS). In the Czech Republic (CZ), currently eighteen RD are screened: phenylketonuria/hyperphenylalaninemia (PKU/HPA), congenital hypothyroidism (CH), congenital adrenal hyperplasia (CAH), cystic fibrosis (CF), medium chain acyl-CoA dehydrogenase deficiency (MCADD), long chain 3-hydroxyacyl-CoA dehydrogenase deficiency (LCHADD), very long chain acyl-CoA dehydrogenase deficiency (VLCADD), carnitine palmitoyl transferase I and II deficiency (CPTID, CPTIID), carnitine-acylcarnitine translocase deficiency (CACTD), maple syrup urine disease (MSUD), glutaric aciduria type I (GA I), isovaleryl-CoA dehydrogenase deficiency (IVA), argininemia (ARG), citrullinemia (CIT), biotinidase deficiency (BTD), cystathionine beta-synthase-deficient homocystinuria (CBSD HCU), and methylenetetrahydrofolate reductase deficiency homocystinuria (MTHFRD HCU). The aim was to analyze the prevalence of RD screened by NBS in CZ. METHODS: We examined the NBS programme in CZ from 1 January 2010 to 31 December 2017, which covered 888,891 neonates. Dried blood spots were primarily analyzed using fluorescence immuno-assay, tandem mass spectrometry and fluorimetry. RESULTS: The overall prevalence of RD among the neonate cohort was 1 : 1,043. Individually, 1 : 2,877 for CH, 1 : 5,521 for PKU/HPA, 1 : 6,536 for CF (1 : 5,887 including false negative patients), 1 : 12,520 for CAH, 1 : 22,222 for MCADD, 1 : 80,808 for LCHADD, 1 : 177,778 for GA I, 1 : 177,778 for IVA, 1 : 222,223 for VLCADD, 1 : 296,297 for MSUD, 1 : 8,638 for BTD, and 1 : 181,396 for CBSD HCU. CONCLUSIONS: The observed prevalence of RD, based on NBS, corresponds to that expected, more precisely it was higher for BTD and lower for MSUD, IVA, CBSD HCU, MCADD and VLCADD. Early detection of rare diseases by means of NBS is an effective secondary prevention tool.

• MeSH
• biologické markery krev   MeSH
• fluorometrie MeSH
• lidé MeSH
• novorozenec MeSH
• novorozenecký screening metody   MeSH
• tandemová hmotnostní spektrometrie MeSH
• vzácné nemoci krev   epidemiologie   MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Neonates with low birthweight (LBW) represent a vulnerable population. This retrospective study analyzed the birth frequency of diseases detected by neonatal screening (NBS) in normal and LBW neonates in the Czech Republic. Between years 2002 and 2016, the number of screened disorders in the Czech Republic gradually increased from two to 13. Prevalence of screened diseases was calculated for cohorts ranging from 777,100 to 1,277,283 neonates stratified by birthweight. Odds ratio of the association of LBW with each disease was calculated and statistical significance was evaluated using the chi-square test or Fisher's exact test, as appropriate. Three diseases were associated with higher risk of prevalence in LBW neonates, namely congenital hypothyroidism (OR 2.50, CI 1.92; 3.25), cystic fibrosis (OR 2.44, CI 1.51; 3.94), and long chain 3-hydroxyacyl-CoA dehydrogenase deficiency (LCHADD) (OR 7.74, CI 2.18; 27.42).Conclusion: Although the underlying mechanisms are not well understood, results can be hypothesized that LBW (respectively prematurity) may lead to the secondary and often transitory hypothyroidism while cystic fibrosis and LCHADD may manifest already prenatally and result into preterm birth and LBW. What is Known: • The percentage of low birthweight (LBW) neonates in the Czech Republic has been increasing. • Previously published studies reported positive association between LBW and congenital hypothyroidism and cystic fibrosis. What is New: • The association between LCHADD and LBW has not yet been described. • LBW can be the first manifestation of cystic fibrosis and LCHADD.

• MeSH
• lidé MeSH
• nemoci novorozenců epidemiologie   MeSH
• novorozenec s nízkou porodní hmotností MeSH
• novorozenec MeSH
• novorozenecký screening normy   MeSH
• porodní hmotnost MeSH
• prevalence MeSH
• retrospektivní studie MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

• Publikační typ
• abstrakt z konference MeSH

AIM: To analyze the prognostic significance of serum and urinary neopterin concentrations in patients with rectal adenocarcinoma treated with (chemo)radiation. PATIENTS AND METHODS: Urinary and serum neopterin and peripheral blood cell count were determined in 49 patients with rectal carcinoma before the start of (chemo)radiation. RESULTS: Neopterin concentrations exhibited a significant inverse correlation with hemoglobin and positive correlation with leukocyte count, platelet count and platelet-to-lymphocyte ratio. Increased serum neopterin concentration was associated with significantly inferior relapse-free survival (RFS) and overall survival. However, a significant association was observed only in 28 patients treated in the neoadjuvant setting. Although increased urinary neopterin was also associated with inferior RFS and overall survival, this was not statistically significant. The neutrophil-to-lymphocyte ratio was also associated with poor prognosis. CONCLUSION: The data presented herein indicate a prognostic significance of serum neopterin concentrations in patients with rectal cancer treated with neoadjuvant chemoradiation.

• MeSH
• chemoradioterapie metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory rekta radioterapie   terapie   MeSH
• neopterin krev   moč   MeSH
• prognóza MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Dosažení patologické kompletní odpovědi po neoadjuvantní chemoterapii je ukazatelem dlouhodobé kontroly onemocnění. V předchozím projektu byla patologická kompletní odpověď pozorována významně častěji u nemocných s nádory expresí HER-2 a bez exprese HER-2 i hormonálních receptorů. Cílem předkládaného projektu je studovat biologickou odpověď na neoadjuvantní chemoterapii. Použity budou standardní režimy chemoterapie používané v jednotlivých centrech a volbu léčby projekt neovlivní. V retrospektivní studii na materiálu nemocných z 2 center budou vyhodnoceny rozdíly tumor infiltrujících lymfocytů u nemocných s různým fenotypem nádoru, bude také studován vliv exprese indoleamin 2,3-dioxygenázy na přítomnost tumor infiltrujících lymfocytů a bude provedeno imunohistochemické zhodnocení fenotypu nádoru. Budou rovněž sledovány změny metabolismu železa a citrulin, ukazatel toxicity léčby.; Pathologic complete response is an indicator of long-term disease control after neoadjuvant chemotherapy in breast carcinoma. In our earlier study, patologic complete response was observed significantly more often in patients with tumors expressing HER-2 or triple negative tumors. The aim of the present study is to study biologic response to neoadjuvant chemotherapy in patients treated with standard regimens that will not be affected by the project. In a retrospective study on material from patients in 2 centers difference in tumor infiltrating lymphocytes in patients with different tumor phenotype as well as the association between indoleamine 2,3-dioxygenase expression and the presence of tumor infiltrating lymphocytes will be evaluated together with tumor phenotype using immunohistochemistry. Alterations of iron metabolism and citrulline, an indicator of intestinal toxicity, will also be assessed.

• MeSH
• alfa-tokoferol MeSH
• citrulin analýza   MeSH
• fenotyp MeSH
• imunohistochemie MeSH
• imunologické faktory MeSH
• indolamin-2,3,-dioxygenasa MeSH
• karcinom prsu in situ MeSH
• kynurenin MeSH
• lymfocyty účinky léků   MeSH
• nádory prsu MeSH
• neoadjuvantní terapie MeSH
• neopterin MeSH
• poruchy metabolismu železa MeSH
• prognóza MeSH
• receptor erbB-2 MeSH
• retrospektivní studie MeSH
• triple-negativní karcinom prsu MeSH
• vitamin A MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• onkologie
• gynekologie a porodnictví
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakt z konference MeSH

Pomocí celoplošného novorozeneckého laboratorního screeningu ze suché kapky krve (SKK) na filtračním papírku jsou od roku 1975 vyhledáváni pacienti s fenylketonurií/hyperfenylalaninemií (PKU/HPA); od roku 1985 s vrozenou hypotyreózou (CH); od roku 2006 s kongenitální adrenální hyperplazií (CAH) a od X/2009 pacienti s cystickou fibrózou (CF) a 9 dalšími dědičnými poruchami metabolismu – leucinózou (MSUD), glutarovou acidurií typu I (GA I), izovalerovou acidurií (IVA), deficitem dehydrogenázy acyl-CoA se středně dlouhým, 3-hydroxyacyl-CoA s dlouhým a acyl-CoA s velmi dlouhým řetězcem (MCADD, LCHADD a VLCADD), deficitem karnitinpalmitoyltransferázy I a II (CPTD I a II) a deficitem karnitinacylkarnitintranslokázy (CACTD). Metodika: V období od 1. ledna 2002 až do 31. prosince 2012 bylo vyšetřeno 1 179 136 novorozenců na PKU//HPA a CH; koncentrace fenylalaninu byla měřena různými metodami a od X/2009 jednotně tandemovou hmotnostní spektrometrií (MS/MS); koncentrace tyreotropinu byla zjišťována pomocí fluoroimunoeseje (FIA). U 752 922 novorozenců byla změřena koncentrace 17-hydroxyprogesteronu pomocí FIA. U 367 114 dětí byl změřen imunoreaktivní trypsin (IRT) pomocí FIA a u 1,05 % novorozenců s nejvyšším IRT bylo vyšetřeno v prvotních SKK 32 a později 50 nejčastějších mutací v genu CFTR s cílem minimalizovat falešnou pozitivitu. U 362 653 novorozenců byly analyzovány metabolity pro záchyt dalších 9 dědičných metabolických poruch pomocí MS/MS. Výsledky: Celkem bylo novorozeneckým screeningem zachyceno 646 pacientů s následně potvrzenou diagnózou. Screeningové prevalence jednotlivých nemocí byly následující CH 1:3562; CAH 1:12 343; CF 1:7060; PKU/HPA 1:7234; MCADD 1:17 269; LCHADD 1:51 808; VLCADD 1:90 663; GA I 1:120 884; MSUD a IVA shodně 1:181 327. Screeningem dosud nebyl zachycen žádný novorozenec s CPTD I, II a CACTD. Efektivita screeningového programu v ČR se zvyšovala, kumulativní četnost záchytů narůstala z 1:2701 v letech 2002–2005 přes 1:2072 v letech 2007–2008 až na 1:1138 po zásadním rozšíření novorozeneckého screeningu (NS) v roce 2009. Četnost falešné pozitivity (FPR) činila 0,540 % , z toho 74 % reprezentuje NS CAH. Závěr: NS je účinný nástroj sekundární prevence vzácných onemocnění s účinnou léčbou. Pro optimalizaci programu se zavádějí druhostupňová vyšetření u malého množství vzorků s cílem snížit falešnou pozitivitu a hledají se možnosti rozšíření spektra screenovaných chorob.

The nationwide newborn screening (NBS) using dried blood spots (DBS) was started in 1975 for phenylketonuria//hyperphenylalaninemia (PKU/HPA); in 1985 for congenital hypothyroidism (CH); in 2006 for congenital adrenal hyperplasia (CAH) and in X/2009 screening for cystic fibrosis (CF) and 9 inherited metabolic diseases – maple syrup urine disease (MSUD), glutaric aciduria type I (GA I), isovaleric aciduria (IVA), medium chain acyl-CoA, long chain 3-hydroxyacyl-CoA and very long chain acyl-CoA dehydrogenase deficiency (MCADD, LCHADD and VLCADD), carnitine palmitoyltransferase I and II deficiency (CPTD I and II) and carnitine-acylcarnitine translocase deficiency (CACTD) was mandated. Methods: From 1st January 2002 through 31st December 2012 a total of 1 179,136 newborns were screened for PKU//HPA and CH; phenylalanine was measured by different methods, since X/2009 solely by tandem mass spectrometry (MS/MS); the levels of thyreotropin were measured by fluoroimmunoassay (FIA). The of 17-hydroxyprogesterone concentrations were determined in 752,922 newborns by FIA. The level of immunoreactive trypsinogen (IRT) was measured in 367,114 newborns by FIA and the CFTR gene (at first 32, later 50 most common mutations) was analysed in original DBS in 1.05% of newborns with the highest IRT levels to minimize false positive results. The spectrum of metabolites for 9 expanded inborn errors of metabolism was analysed in 362,653 newborns by MS/MS. Results: NS detected in total 646 patients with subsequently confirmed diagnosis. Screening prevalence were as follows PKU/HPA 1:7,234; CH 1:3,562; CAH 1:12,343; CF 1:7,060; MCADD 1:17,269; LCHADD 1:51,808; VLCADD 1:90,663; GA I 1:120,884; MSUD and IVA both 1:181,327. No case of CPTD I and CPTDII/CACTD was detected. Efficacy of NBS increased with the expansion of diseases in the programme: detection rate rose from 1:2,701 in 2002–2005 over 1:2,072 in 2007–2008 to 1:1,138 after last expansion of NBS in 2009. False positive rate (FPR) was 0.540%, of which 74% represent results of CAH screening. Conclusion: NBS is an effective approach for presymptomatic detection of serious rare diseases. Further optimization is a subject of current research including the introduction of additional analytical tests (second tiers) to reduce FPR and by further expanding of screened disorders.

• MeSH
• biochemická analýza krve MeSH
• falešně negativní reakce MeSH
• falešně pozitivní reakce MeSH
• fenylketonurie MeSH
• kongenitální hypotyreóza MeSH
• krev MeSH
• lidé MeSH
• novorozenec MeSH
• novorozenecký screening * MeSH
• plošný screening * MeSH
• porodní hmotnost MeSH
• prevalence MeSH
• vrozené vady diagnóza   MeSH
• vzácné nemoci diagnóza   MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• Publikační typ
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH