End-stage kidney disease is preferably treated by kidney transplantation. The suboptimal function of the allograft often results in misbalances in kidney-controlled processes and requires long-term monitoring of allograft function and viability. As the kidneys are organs with a very high metabolomic rate, a metabolomics approach is suitable to describe systematic changes in post-transplant patients and has great potential for monitoring allograft function, which has not been described yet. In this study, we used blood plasma samples from 55 patients after primary kidney transplantation identically treated with immunosuppressants with follow-up 50 months in the mean after surgery and evaluated relative levels of basal plasma metabolites detectable by NMR spectroscopy. We were looking for the correlations between circulating metabolites levels and allograft performance and allograft rejection features. Our results imply a quantitative relationship between restricted renal function, insufficient hydroxylation of phenylalanine to tyrosine, lowered renal glutamine utilization, shifted nitrogen balance, and other alterations that are not related exclusively to the metabolism of the kidney. No link between allograft function and energy metabolism can be concluded, as no changes were found for glucose, glycolytic intermediates, and 3-hydroxybutyrate as a ketone body representative. The observed changes are to be seen as a superposition of changes in the comprehensive inter-organ metabolic exchange, when the restricted function of one organ may induce compensatory effects or cause secondary alterations. Particular differences in plasma metabolite levels in patients with acute cellular and antibody-mediated allograft rejection were considered rather to be related to the loss of kidney function than to the molecular mechanism of graft rejection since they largely follow the alterations observed by restricted allograft function. In the end, we showed using a simple mathematical model, multilinear regression, that the basal plasmatic metabolites correlated with allograft function expressed by the level of glomerular filtration rate (with creatinine: p-value = 4.0 × 10-26 and r = 0.94, without creatinine: p-value = 3.2 × 10-22 and r = 0.91) make the noninvasive estimation of the allograft function feasible.
- Publikační typ
- časopisecké články MeSH
Cieľ: Voľné ľahké reťazce kappa (kappa free light chains; k-fLC) v likvore sú perspektívnym novým markerom intratekálnej syntézy imunoglobulínov (Ig) u pacientov s RS, pričom publikované referenčné hodnoty nie sú jednotné. Cieľom našej práce bolo analyzovať diagnostickú hodnotu k-fLC (hladiny v likvore, kvocientu, indexu) a rutinne vyšetrovaných markerov (IgG index, IgGReiber) v diagnostike RS, v predikcii pozitivity oligoklonálnej skladby (oligoclonal bands; OCB) a u OCB negatívnych pacientov. Metódy: Párované vzorky séra a likvoru boli analyzované v súbore 46 pacientov s RS a 63 kontrolných jedincov s nezápalovými a inými zápalovými ochoreniami CNS. Koncentrácia k-fLC bola meraná imunoturbidimetricky (SPA PLUS®, Freelite®). Pre jednotlivé markery a ich cut-off hodnoty, určené s použitím Youdenovho indexu, boli porovnané ROC krivky, diagnostické senzitivity a špecificity. Výsledky: Optimálne cut-off hodnoty boli: 0,76 pre IgG index; 0,89 mg/l pre IgGReiber; 1,08 mg/l pre hladinu k-fLC v likvore; 0,0994 pre k-fLC kvocient; 18,15 pre k-fLC index. K odlíšeniu RS od control boli zistené najvyššie kombinované diagnostické senzitivity/špecificity pre k-fLC index (0,76/0,98), nasledované k-fLC kvocientom (0,76/0,97), hladinou k-fLC v likvore (0,76/0,95), intratekálnou syntézou IgGReiber (0,70/0,91) a IgG indexom (0,65/0,89). Senzitivita a špecificita OCB vyšetrenia bola 0,83 a 1,00. V predikcii OCB pozitivity malo použitie nových k-fLC markerov excelentnú diagnostickú využiteľnosť so senzitivitou/špecificitou: 0,92/0,99 pre k-fLC index; 0,92/0,97 pre k-fLC kvocient; 0,92/0,96 pre hladinu k-fLC v likvore, v porovnaní s 0,76/0,89 pre IgG index a 0,76/0,86 pre IgGReiber. U OCB negatívnych RS pacientov s použitím navrhnutých cut-off bola najvyššia diagnostická senzitivita (0,38) detekovaná pre IgGReiber, nasledovaná senzitivitou 0,13 pre IgG index a prekvapivo 0,00 senzitiviou pre k-fLC markery. Záver: V diagnostike RS a pri predikcii OCB pozitivity sme detekovali pre všetky analyzované k-fLC markery lepšie diagnostické senzitivity a špecificity v porovnaní s rutinne používaným IgG indexom a IgGReiber, tieto však nedosiahli senzitivitu a špecificitu OCB vyšetrenia. Pre dôkaz intratekálnej syntézy Ig u pacientov so suspektnou RS odporúčame paralelné vyšetrenie OCB spolu s použitím k-fLC indexu a kvocientu, a to predovšetkým ak OCB nález je nejednoznačný alebo negatívny.
Aim: Kappa free light chains (-fLC) in cerebrospinal fl uid (CSF) were suggested as promising novel markers of intrathecal immunoglobulin (Ig) synthesis in MS patients, with reported discrepant reference values. We aimed to analyse the diagnostic accuracy of k-fLC (CSF concentration, quotient, index) and routinely used markers (IgG index, IgGReiber) in MS diagnostics, in prediction of oligoclonal bands (OCB) positivity and in OCB negative patients. Methods: Serum/CSF paired samples were analysed from 46 MS patients and 63 controls with non-inflammatory and non-MS inflammatory diseases of CNS. The concentrations of k-fLC were measured immunoturbidimetrically (SPA PLUS®, Freelite®). The ROC curves, diagnostic sensitivities and specificities were analysed with respect to the cut-offs, which were determined using Youden’s index. Results: The optimal cut-off values were defined as: 0.76 for IgG index; 0.89 mg/L for IgGReiber; 1.08 mg/L for CSF k-fLC level; 0.0994 for k-fLC quotient; 18.15 for k-fLC index. To distinguish MS patients from controls, the highest combined sensitivities/specificities were observed for k-fLC index (0.76/0.98), followed by k-fLC quotient (0.76/0.97), CSF k-fLC level (0.76/0.95), IgGReiber (0.70/0.91) and IgG index (0.65/0.89). The OCB detection showed the sensitivity 0.83 and the specificity 1.00. In prediction of OCB positivity, the novel k-fLC markers showed excellent diagnostic accuracy with sensitivities/specificities: 0.92/0.99 for k-fLC index, 0.92/0.97 for k-fLC quotient, and 0.92/0.96 for CSF k-fLC level, compared to 0.76/0.89 for IgG index and 0.76/0.86 for IgGReiber. Applying our cut-offs in OCB negative MS patients, the highest diagnostic sensitivity (0.38) was found for IgGReiber, followed by 0.13 for IgG index, and surprisingly by 0.00 for all k-fLC markers. Conclusion: In MS diagnostics and in prediction of OCB positivity, all novel k-fLC markers showed better sensitivities and specificities than routinely used IgG index and IgGReiber, but did not reach those of OCBs. To proove the intrathecal Ig synthesis in patients with suspected MS, we recommend to use k-fLC index and quotient along with OCB, especially if OCB findings are equivocal or negative.
Cieľ: Včasné rozpoznanie sclerosis multiplex (SM) pomáha začať liečbu pacientov skôr, a tak oddialiť progresiu ochorenia. Urobili sme analýzu metabolitov cerebro-spinálneho likvoru (cerebrospinal fluid; CSF), s cieľom zistiť prediktory včas, a tak oddialiť SM. Metódy: Do štúdie bolo zaradených 56 jedincov s podozrením na SM, pred začatím akejkoľvek liečby. Z nich bolo 28 diagnostikovaných ako definitívna SM, u 17 pacientov sme zistili klinicky izolovaný syndróm (clinically isolated syndrome; CIS) podľa McDonaldových kritérií z roku 2010, v 11 prípadoch sa jednalo o iné demyelinizačné ochorenie CNS (DEM). Kontrolnú skupinu (CON) tvorili 29 jedinci, ktorí nemali dokázané žiadne ochorenie CNS. Na meranie metabolitov CSF bola použitá protonová nukleárna magnetická rezonančná spektroskopia. Výsledky: Glutamín, ktorý koreloval s Expanded Disability Status Scale (EDSS), bol jediným metabolitom, ktorý dokázal odlíšiť CIS, SM, DEM a CON. Valín, leucín, isoleucín, znížené u CIS a SM v porovnaní s CON, sa neodlišovali od DEM. Hladiny citrátu v CSF špecifikovali SM a CIS oproti DEM, ale nepomohli v rozlíšení CIS a SM. Citrát ukazoval signifikantné korelácie s vekom, dľžkou trvania ochorenia a EDSS u SM pacientov. Acetát, aceton, pyruvát, formát, histidin v CSF neboli signifikantnými prediktormi SM alebo CIS, hoci korelovali s niektorými vybranými premennými. Záver: Táto práca ukazuje prediktívnu úlohu glutamínu v CSF v stanovení diagnózy SM od jej včasných štádií, vypichujúc tak dôležitú úlohu glutamát/glutamínového cyklu v patogenéze SM. Ďalší potenciálny prediktor SM bol citrát. Ďalšie metabolity neboli identifikované ako senzitívne CSF markery SM.
Aim: Early recognition of multiple sclerosis (MS) allows patients to begin treatment earlier and delay disease progression. We performed an analysis of cerebrospinal fluid (CSF) metabolites to find early predictors of MS. Methods: We included 56 participants with suspected MS before any treatment. Out of those, 28 patients were diagnosed with definite MS, 17 with clinically isolated syndrome (CIS) according to McDonald 2010 criteria, and 11 with other demyelinating diseases (DEM) of the CNS. The control group (CON) included 29 participants without any confirmed CNS disease. Proton nuclear magnetic resonance spectroscopy was used to measure CSF metabolites. Results: Glutamine, correlating with Expanded Disability Status Scale (EDSS), was the only metabolite capable to distinguish between CIS and MS, DEM, and CON. Valine, leucine, isoleucine, decreased in CIS and MS when compared with CON, did not differ from DEM. Citrate CSF levels specified MS and CIS against DEM but did not help to distinguish between CIS and MS. Citrate showed significant correlations with age, disease duration, and EDSS in MS patients. Acetate, acetone, pyruvate, formate and histidine CSF levels were not significant predictors of MS or CIS, although they correlated with selective variables. Conclusion: This work shows the predictive role of CSF glutamine in diagnosing MS since its early stages, pinpointing an important role of the glutamate/glutamine cycle in MS pathogenesis. Another potential predictor of MS was citrate. Other metabolites were not identified as sensitive CSF markers of MS.
- MeSH
- demyelinizační nemoci diagnostické zobrazování MeSH
- klinická studie jako téma MeSH
- lidé MeSH
- magnetická rezonanční spektroskopie metody MeSH
- metabolomika MeSH
- mozkomíšní mok diagnostické zobrazování MeSH
- roztroušená skleróza * diagnóza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Oxidative stress and decline in cellular redox regulation have been hypothesized to play a key role in cardiovascular aging; however, data on antioxidant and redox regulating systems in the aging heart are controversial. The aim of the present study was to examine the effect of aging on critical antioxidant enzymes and two major redox-regulatory systems glutathione (GSH) and thioredoxin (Trx) system in hearts from adult (6-month-old), old (15-month-old), and senescent (26-month-old) rats. Aging was associated with a non-uniform array of changes, including decline in contents of reduced GSH and total mercaptans in the senescent heart. The activities of Mn-superoxide dismutase (SOD2), glutathione peroxidase (GPx), glutathione reductase (GR), and thioredoxin reductase (TrxR) exhibited an age-related decline, whereas catalase was unchanged and Cu,Zn-superoxide dismutase (SOD1) displayed only slight decrease in old heart and was unchanged in the senescent heart. GR, Trx, and peroxiredoxin levels were significantly reduced in old and/or senescent hearts, indicating a diminished expression of these proteins. In contrast, SOD2 level was unchanged in the old heart and was slightly elevated in the senescent heart. Decline in GPx activity was accompanied by a loss of GPx level only in old rats, the level in senescent heart was unchanged. These results indicate age-related posttranslational protein modification of SOD2 and GPx. In summary, our data suggest that changes are more pronounced in senescent than in old rat hearts and support the view that aging is associated with disturbed redox balance that could alter cellular signaling and regulation.
- MeSH
- antioxidancia analýza metabolismus MeSH
- glutathionperoxidasa analýza metabolismus MeSH
- krysa rodu rattus MeSH
- myokard chemie enzymologie MeSH
- oxidace-redukce MeSH
- potkani Wistar MeSH
- stárnutí metabolismus MeSH
- superoxiddismutasa metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
Increased level of homocysteine (hHcy) in plasma is an accompanying phenomenon of many diseases, including a brain stroke. This study determines whether hyperhomocysteinemia (which is a risk factor of brain ischemia) itself or in combination with ischemic preconditioning affects the ischemia-induced neurodegenerative changes, generation of reactive oxygen species (ROS), lipoperoxidation, protein oxidation, and activity of antioxidant enzymes in the rat brain cortex. The hHcy was induced by subcutaneous administration of homocysteine (0.45 μmol/g body weight) twice a day in 8 h intervals for 14 days. Rats were preconditioned by 5 min ischemia. Two days later, 15 min of global forebrain ischemia was induced by four vessel's occlusion. The study demonstrates that in the cerebral cortex, hHcy alone induces progressive neuronal cell death and morphological changes. Neuronal damage was associated with the pro-oxidative effect of hHcy, which leads to increased ROS formation, peroxidation of lipids and oxidative alterations of cortical proteins. Ischemic reperfusion injury activates degeneration processes and de-regulates redox balance which is aggravated under hHcy conditions and leads to the augmented lipoperoxidation and protein oxidation. If combined with hHcy, ischemic preconditioning could preserve the neuronal tissue from lethal ischemic effect and initiates suppression of lipoperoxidation, protein oxidation, and alterations of redox enzymes with the most significant effect observed after prolonged reperfusion. Increased prevalence of hyperhomocysteinemia in the Western population and crucial role of elevated Hcy level in the pathogenesis of neuronal disorders makes this amino acid as an interesting target for future research. Understanding the multiple etiological mechanisms and recognition of the co-morbid risk factors that lead to the ischemic/reperfusion injury and ischemic tolerance is therefore important for developing therapeutic strategies in human brain stroke associated with the elevated level of Hcy.
- MeSH
- hyperhomocysteinemie komplikace enzymologie patologie MeSH
- krysa rodu rattus MeSH
- oxidace-redukce MeSH
- oxidační stres fyziologie MeSH
- peroxidace lipidů fyziologie MeSH
- potkani Wistar MeSH
- přivykání k ischémii trendy MeSH
- reaktivní formy kyslíku metabolismus MeSH
- reperfuzní poškození enzymologie patologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Major depressive disorder (MDD) is a main public health concern worldwide. Despite extensive investigations, the exact mechanisms responsible for MDD have not been identified. Epidermal growth factor (EGF) and insulin growth factor binding protein-3 (IGFBP-3) are involved in brain function. Tumour suppressor protein p53 is widely involved in neuronal death in response to different forms of acute insults and neurological disorders. The present study focuses on the possible associations of the single-nucleotide polymorphisms (SNP) of EGF A61G (rs4444903), IGFBP-3 C32G (rs2854746) and TP53 G72C (rs1042522) genes with MDD risk in the Slovak population. METHODS: The present case-control association study was carried out in 111 confirmed MDD patients and 207 healthy subjects. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism methods. RESULTS: Logistic regression analysis showed no association between SNPs of selected genes and MDD risk in the Slovak population. However, the stratification of individuals by gender revealed that males carrying IGFBP-3 G alleles (G32G or GG) had marginally increased risk for developing MDD as compared to CC homozygous males (p=0.09). In women, inverse association was observed between SNP rs1042522 and MDD risk (p=0.04 for recessive model). CONCLUSION: Our results suggest the protective effect of minor allele 72C of TP53 gene towards MDD. The disruption of mechanisms involved in cell survival and death regulation may be involved in pathophysiology of MDD.
- MeSH
- alely MeSH
- depresivní porucha unipolární genetika MeSH
- dospělí MeSH
- epidermální růstový faktor genetika MeSH
- genetická predispozice k nemoci MeSH
- genotyp MeSH
- geny p53 genetika MeSH
- IGFBP-3 genetika MeSH
- jednonukleotidový polymorfismus * MeSH
- lidé středního věku MeSH
- lidé MeSH
- polymerázová řetězová reakce MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- studie případů a kontrol MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Slovenská republika epidemiologie MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
Several neurodegenerative conditions, such as Alzheimer's disease and Parkinson's disease, or vascular dementia and cognitive impairment, are associated with mild hyperhomocysteinemia. Hyperhomocysteinemia is defined as an increase of the homocysteine (Hcy) level beyond 10 microM. Although the adverse effect of Hcy on neurons is well documented, knowledge about the impact of this amino acid on glial cells is missing. Therefore, with the aim to evaluate the neurotoxic properties of Hcy on glial cells, we used a glioblastoma cell line as a study model. The viability of cells was assayed biochemically and cytologically. At a concentration around 50 microM in the culture medium D,L-Hcy induced cell death. It is noteworthy that Hcy induces cell death of human glial cells at concentrations encountered during mild hyperhomocysteinemia. Therefore, we propose that Hcy-induced impairment of neuronal functions along with damage of glial cells may contribute to the etiopathogenesis of neurodegenerative diseases associated with hyperhomocysteinemia.
- MeSH
- buněčná smrt účinky léků fyziologie MeSH
- homocystein farmakologie MeSH
- kultivované buňky MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- neuroglie účinky léků fyziologie MeSH
- neurony účinky léků fyziologie MeSH
- viabilita buněk účinky léků fyziologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
