OBJECTIVES: Both dimethylarginines are widely bound to chronic kidney disease (CKD). This study was focused to validate published LC-MS/MS method and compared the measured data with an immunoassay. DESIGN AND METHODS: The analysis was performed on a Dionex UltiMate 3000 UHPLC-Standard (Thermo Fisher Scientific, Waltham, Massachusetts, USA) with an amaZon SL ion trap (Bruker, Billerica, Massachusetts, USA). Comparison was evaluated by using Passing Bablok regression and Bland Altman plot. Healthy volunteers (n = 40) were used for validation and as control group to patients group (n = 40) with different stages of CKD. RESULTS: The results in healthy controls determined by the LC-MS/MS (ELISA) method were 0.52 ± 0.0892 with 95 % CI: 0.49-0.55 (0.61 ± 0.1213 with 95 % CI: 0.57-0.64) μmol/L for AD MA and 0.56 ± 0.0810 with 95 % CI: 0.53-0.58 (0.62 ± 0.0752 with 95 % CI: 0.57-0.65) μmol/L for SDMA. In the same way, the patient group values determined by the LC-MS/MS (ELISA) method were 0.82 ± 0.1604 with 95 % CI: 0.75-0.88 (1.06 ± 0.3002 with 95 % CI: 0.94-1.19) μmol/L and 2.14 ± 0.8778 with 95 % CI: 1.47-2.58 (1.65 ± 0.5160 with 95 % CI: 1.40-1.98) μmol/L for ADMA and SDMA, respectively. The correlation between the methods, expressed as the Spearman correlation coefficient (R), was 0.858 (0.8059) for ADMA (p < 0.0001) and 0.895 (0.9607) for SDMA (p < 0.0001). CONCLUSIONS: ADMA levels determined by the immunoassay were almost 30 % overestimated, in contrast to SDMA levels, which were 3 % underestimated. According to our findings, a better correlation could be obtained by simple sample dilution.

• Publikační typ
• časopisecké články MeSH

• Publikační typ
• abstrakt z konference MeSH

The resistance to carbapenems is usually mediated by enzymes hydrolyzing β-lactam ring. Recently, an alternative way of the modification of the antibiotic, a β-lactone formation by OXA-48-like enzymes, in some carbapenems was identified. We focused our study on a deep analysis of OXA-48-like-producing Enterobacterales, especially strains showing poor hydrolytic activity. In this study, well characterized 74 isolates of Enterobacterales resistant to carbapenems were used. Carbapenemase activity was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), liquid chromatography/mass spectrometry (LC-MS), Carba-NP test and modified Carbapenem Inactivation Method (mCIM). As meropenem-derived β-lactone possesses the same molecular weight as native meropenem (MW 383.46 g/mol), β-lactonization cannot be directly detected by MALDI-TOF MS. In the spectra, however, the peaks of m/z = 340.5 and 362.5 representing decarboxylated β-lactone and its sodium adduct were detected in 25 out of 35 OXA-48-like producers. In the rest 10 isolates, decarboxylated hydrolytic product (m/z = 358.5) and its sodium adduct (m/z = 380.5) have been detected. The peak of m/z = 362.5 was detected in 3 strains co-producing OXA-48-like and NDM-1 carbapenemases. The respective signal was identified in no strain producing class A or class B carbapenemase alone showing its specificity for OXA-48-like carbapenemases. Using LC-MS, we were able to identify meropenem-derived β-lactone directly according to the different retention time. All strains with a predominant β-lactone production showed negative results of Carba NP test. In this study, we have demonstrated that the strains producing OXA-48-like carbapenemases showing false-negative results using Carba NP test and MALDI-TOF MS preferentially produced meropenem-derived β-lactone. We also identified β-lactone-specific peak in MALDI-TOF MS spectra and demonstrated the ability of LC-MS to detect meropenem-derived β-lactone.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální proteiny * analýza   MeSH
• beta-laktamasy analýza   MeSH
• Enterobacteriaceae * MeSH
• karbapenemy farmakologie   MeSH
• meropenem farmakologie   MeSH
• mikrobiální testy citlivosti MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: Asthma as a chronic inflammatory disorder has been suggested as a risk factor for endothelial dysfunction (ED), but studies on the association between asthma and cardiovascular disease (CVD) risk are limited. BACKGROUND: We assessed associations of ED with the severity of asthma, eosinophilic inflammation, lung function, and asthma control. METHODS: 52 young asthmatics (median age of 25.22 years) and 45 healthy individuals were included. Demographic, clinical, and laboratory findings were recorded. We evaluated microvascular responsiveness by recording the reactive hyperemia index (RHI) indicating post-occlusive peripheral endothelium-dependent changes in vascular tone using the Itamar Medical EndoPAT2000. VCAM-1, ADMA, high-sensitive CRP (hsCRP), and E-selectin were measured. RESULTS: Asthmatics had considerably lower RHI values (p < 0.001) with a dynamic decreasing trend by asthma severity and higher hsCRP levels (p < 0.001). A substantial increase in hsCRP and E-selectin with asthma severity (p < 0.05) was also observed. We confirmed a higher body mass index (BMI) in asthmatics (p < 0.001), especially in women and in severe asthma. CONCLUSIONS: We demonstrated the progression of CVD in asthmatics and the association of the ongoing deterioration of ED with the inflammatory severity, suggesting that the increased risk of CVD in young asthmatics is dependent on disease severity. The underlying mechanisms of risk factors for CVD and disease control require further study.

• Publikační typ
• časopisecké články MeSH

Exposure to endocrine disruptors such as bisphenols, can lead to and be the explanation for idiopathic infertility. In our study, we assessed the effect of exposure to bisphenol S (BPS) via breast milk on the testicular tissue health of adult male mice. Lactating dams were exposed to BPS through drinking water (0.216 ng g bw/day and 21.6 ng g bw/day) from post-natal day 0-15. Although there was no significant difference in testicular histopathology between the control and experimental groups, we observed an increase in the number of tight and gap junctions in the blood-testis barrier (BTB) of adult mice after lactation BPS exposure. Moreover, there was an increase in oxidative stress markers in adult testicular tissue of mice exposed via breast milk. Our lactation model indicates that breast milk is a route of exposure to an endocrine disruptor that can be responsible for idiopathic male infertility through the damage of the BTB and weakening of oxidative stress resistance in adulthood.

• MeSH
• benzhydrylové sloučeniny toxicita   MeSH
• endokrinní disruptory * toxicita   MeSH
• laktace * MeSH
• myši MeSH
• sulfony toxicita   MeSH
• testis MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION AND OBJECTIVES: Cardiac involvement in systemic sclerosis (SSc) patients affects mortality. Cardiac magnetic resonance (CMR) is capable of detecting structural changes, including diffuse myocardial fibrosis that may develop over time. Our aim was to evaluate myocardial structure and function changes using CMR in patients with SSc without known cardiac disease during a 5-year follow-up and find possible correlations with selected biomarkers. METHODS: A total of 25 patients underwent baseline and follow-up CMR examinations according to a pre-specified protocol. Standard biochemistry, five biomarkers (hsTnI, NT-proBNP, galectin-3, sST2, and GDF-15), and disease-specific functional parameters enabling the classification of disease severity were also measured. RESULTS: After five years, no patient suffered from manifest heart disease. Mean extracellular volume (ECV) and T1 mapping values did not change significantly (p ≥ 0.073). However, individual increases in native T1 time and ECV correlated with increased galectin-3 serum levels (r = 0.56; p = 0.0050, and r = 0.71; p = 0.0001, respectively). The progression of skin involvement assessed using the Rodnan skin score and a decrease in the diffusing capacity of the lungs were associated with increased GDF-15 values (r = 0.63; p = 0.0009, and r = -0.51; p = 0.011, respectively). CONCLUSIONS: During the 5-year follow-up, there was no new onset of heart disease observed in patients with SSc. However, in some patients, CMR detected progression of sub-clinical myocardial fibrosis that significantly correlated with elevated galectin-3 levels. GDF-15 values were found to be associated with disease severity progression.

• Publikační typ
• časopisecké články MeSH

Cíl studie: Cílem studie bylo zavedení metody stanovení betainu a dimethylglycinu metodou HPLC s UV detekcí ve vzorcích lidské plazmy. Dále stanovení těchto analytů u pacientů s metabolickým syndromem a prediabetem a zhodnocení naměřených výsledků vzhledem ke kontrolní skupině.Typ studie: retrospektivní.Název a sídlo pracoviště:Ústav klinické biochemie a hematologie Lékařské fakulty UK a Fakultní nemocnice, Alej Svobody 80, 304 60 Plzeň.Materiál a metody: Metoda HPLC s UV detekcí byla zavedena na kapalinovém chromatografu Thermo Separation Products Spectra SYSTEM (Thermo Fisher Scientific, Waltham, MA, USA). Podmínky separace: složení mobilní fáze 90 % acetonitrilu, 10 % redestilované vody a 22 mmol/L cholinu (isokratická eluce); průtok mobilní fáze 1,5 ml/min; teplota analýzy 33 °C; UV detekce při 254 nm; kolona SUPELCOSILTM LC-SCX o velikosti částic 5 μm. Pro optimalizaci metody stanovení byly využity vzorky plazmy od 30 dárců z transfuzního oddělení Fakultní nemocnice v Plzni. Soubor obsahoval 15 mužů a žen ve věkovém rozmezí od 39 do 61 let. Soubor pacientů s metabolickým syndromem a prediabetem obsahoval 12 mužů a 8 žen ve věkovém rozmezí od 36 do 67 let. Výsledky: Hodnoty preciznosti v sérii vyjádřené variačním koeficientem byly 10,6 % a 11,8 % pro betain a dimethylglycin, v případě mezilehlé preciznosti 7,1 % a 12,3 %. Výtěžnost metody byla 97,8 % pro betain a 97,7 % pro dimethylglycin. Plaz-matické koncentrace betainu a dimethylglycinu u kontrolní skupiny byly 135,8 ± 47,4 μmol/L a 4,7 ± 1,3 μmol/L. U pacientů s metabolickým syndromem a prediabetem byly plazmatické koncentrace 100,9 ± 40,9 μmol/L (statisticky významně nižší než u kontrol, p < 0,005) a 10,3 ± 6,2 μmol/L (statisticky významně vyšší než u kontrol, p < 0,01). Plazmatická koncentrace homocysteinu byla u pacientů s metabolickým syndromem 14,4 ± 5,0 μmol/L (p < 0,001), u kontrolní skupiny 10,0 ± 3,5 μmol/L.Závěr: U pacientů s metabolickým syndromem a prediabetem jsme zaznamenali ve shodě s jinými autory ve srovnání s kontrolní skupinou dárců krve nižší plazmatickou hladinu betainu a vyšší hladinu dimethylglycinu a homocysteinu. Tyto změny by mohly mít vztah k rozvoji nealkoholické steatózy jater i ke zvýšenému kardiovaskulárnímu riziku u osob s metabolickým syndromem HPLC.

Objective: The aim of our study was the implementation of HPLC method with UV detection for betaine and dimethylglycine in human plasma samples. Then quantification of these analytes in patients with metabolic syndrome and prediabetes and compare results between them and the control group. Design: retrospective. Settings: Department of Clinical Biochemistry and Hematology, Faculty of Medicine, Charles University and Faculty Hospital, Alej Svobody 80, 304 60 Pilsen (Czech Republic). Material and methods: We developed the HPLC method on liquid chromatograph Thermo Separation Products Spectra SYSTEM (Thermo Fisher Scientific, Waltham, MA, USA) with these chromatographic conditions: mobile phase consisting of 90 % acetonitrile, 10 % water and 22 mmol/L choline; flow rate 1.5 ml/min; temperature 33 °C; UV detection in the wavelength 254 nm; column SUPELCOSILTM LC-SCX with particle size size 5 μm. For implementation of method we used plasma samples of blood donors from transfusion department Faculty Hospital in Pilsen. The group consisted of 15 males and 15 females in the range of age of 39 to 61 years. The group with metabolic syndrome and prediabetes contained 12 males and 8 females in the range of age 36 to 67 years. Results: Values of precision expressed as coefficient of variation were 10.6 % and 11.8 % for betaine and dimethylglycine. Intermediate precisions were 7.1 % and 12.3 %. Recovery of both analytes were 97.8 % for betaine and 97.7 % for dimethylglycine. Plasmatic concentrations of betaine and dimethylglycine in the control group were 135.8 ± 47.4 μmol/L and 4.7 ± 1.3 μmol/L. Plasmatic concentrations in patients with metabolic syndrome and prediabetes were 100.9 ± 40.9 μmol/L (significantly lower than in control group, p < 0.005) and 10.3 ± 6.2 μmol/L (significantly higher than in control group, p < 0.01). Plasma concentration of homocysteine in patients with metabolic syndrome was 14.4 ± 5.0 μmol/L (p < 0.001), in the control group 10.0 ± 3.5 μmol/L.Conclusion: Patients with metabolic syndrome and prediabetes had lower plasma concentration of betaine and higher plasma level of dimethylglycine and homocysteine. These changes could be in relation with the development of non-alcoholic fatty liver disease and increased cardiovascular risk in these patients.

• Klíčová slova
• dimethylglycin,
• MeSH
• betain * analýza   MeSH
• klinická studie jako téma MeSH
• lidé MeSH
• metabolický syndrom MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• lidé MeSH