Nucleoside diphosphate kinases (NDPK) are key enzymes involved in the intracellular nucleotide maintenance in all living organisms, especially in trypanosomatids which are unable to synthesise purines de novo. Four putative NDPK isoforms were identified in the Trypanosoma cruzi Chagas, 1909 genome but only two of them were characterised so far. In this work, we studied a novel isoform from T. cruzi called TcNDPK3. This enzyme presents an atypical N-terminal extension similar to the DM10 domains. In T. cruzi, DM10 sequences targeted other NDPK isoform (TcNDPK2) to the cytoskeleton, but TcNDPK3 was localised in glycosomes despite lacking a typical peroxisomal targeting signal. In addition, TcNDPK3 was found only in the bloodstream trypomastigotes where glycolytic enzymes are very abundant. However, TcNDPK3 mRNA was also detected at lower levels in amastigotes suggesting regulation at protein and mRNA level. Finally, 33 TcNDPK3 gene orthologs were identified in the available kinetoplastid genomes. The characterisation of new glycosomal enzymes provides novel targets for drug development to use in therapies of trypanosomatid associated diseases., María de los Milagros Cámara, León Bouvier, Chantal Reigada, Fabio A. Digirolamo, Melisa Sayé, Claudio A. Pereira., and Obsahuje bibliografii
Cíle studie: 1. Zhodnocení buněčností likvoru (CSF, z angl. cerebrospinal fluid = mozkomíšní mok), koncentrací glukózy v CSF, hodnot glukózového kvocientu (Q glu. ), koncentrací laktátu v CSF a hodnot koeficientu energetické bilance (KEB) coby ukazatelů intenzity zánětlivého procesu v centrálním nervovém systému (CNS) ve skupinách pacientů bez postižení CNS, pacientů s mírnými serózními záněty CNS neinfekční etiologie, pacientů se serózními záněty CNS infekční etiologie a pa- cientů s purulentními záněty CNS s extracelulárními bakteriemi v patogenezi. 2. Porovnání informačního potenciálu parametrů energetického metabolismu glukózy v likvorovém kompartmentu v soubo - ru vyšetřených pacientů, tzn. koncentrací glukózy v CSF, hodnot Q glu. , koncentrací laktátu v CSF a hodnot KEB. Typ studie: Retrospektivní studie. Materiál a metody: Vyšetřili jsme 133 vzorků CSF pacientů bez postižení CNS, 227 vzorků CSF pacientů s mírnými serózními záněty CNS s intrathekální syntézou imunoglobulinů neinfekční etiologie, 208 vzorků CSF pacientů se serózními záněty CNS infekční příčiny a 140 vzorků CSF pacientů s purulentními záněty CNS s extracelulárními bakteriemi v patogenezi. Oblastí našeho zájmu byly buněčnost CSF, koncentrace glukózy v CSF, hodnoty Q glu. , koncentrace laktátu v CSF a hodnoty KEB. Pro statistické hodnocení vybraných parametrů jsme použili D’Agostinův omnibus test, Kruskalův-Wallisův test s následnou post hoc analýzou Dunnovou metodou, Spearmanův korelační koeficient a multinomiální logistickou regresní analýzu. Výsledky: V likvorovém kompartmentu pacientů bez postižení CNS jsme nezjistili žádné změny na úrovni buněčností CSF ani změny energetické. V likvorovém kompartmentu pacientů s mírnými serózními záněty CNS neinfekční etiologie byly pouze v některých případech patrny zvýšené buněčnosti CSF a jisté odchylky hodnot Q glu. , koncentrací laktátu v CSF a hod - not KEB od normálu. V likvorovém kompartmentu pacientů se serózními záněty CNS infekční etiologie byly pozorovány projevy nápadně zvýšené intenzity zánětlivého procesu v likvorovém kompartmentu především na úrovni buněčností CSF, koncentrací laktátu v CSF a hodnot KEB. Vysoká intenzita purulentního zánětlivého procesu bakteriální etiologie v CNS se výrazně projevila na úrovni všech sledovaných parametrů. Nejtěsnější vztah jsme v našem souboru pacientů zjistili mezi buněčnostmi CSF a hodnotami KEB ( ρ = -0,770), následoval vztah mezi buněčnostmi CSF a koncentracemi laktátu v CSF ( ρ = 0,734), hodnotami Q glu. ( ρ = -0,676) a koncentracemi glukózy v CSF ( ρ = -0,544). Využitelnost uvedených parametrů k predikci intenzity zánětlivého procesu v CNS jsme ověřili pomocí multinomiální logistické regresní analýzy. Nejvyšší pre - dikční potenciál měly buněčnost CSF se 71,9 % a KEB se 71,6 % správně zařazených pacientů. Následovaly koncentrace laktátu v CSF s 64,7 %, Q glu. s 58,8 % a koncentrace glukózy v CSF s 54,7 % správně klasifikovaných pacientů. Závěr: Získané výsledky podpořily vhodnost využití úrovní buněčností CSF, koncentrací glukózy v CSF, hodnot Q glu. , kon - centrací laktátu v CSF a hodnot KEB k diagnostice postižení a monitorování intenzity zánětů v CNS. Navíc umožnily vytvořit pořadí energetických parametrů v likvorovém kompartmentu pacientů našeho souboru podle úrovně jejich informačního potenciálu. Nejpříznivějších výsledků pro hodnocení intenzity zánětlivého procesu v CNS tak bylo dosaženo v případě hod - not KEB, následovaly je koncentrace laktátu v CSF a hodnoty Q glu. a nejméně příznivých výsledků jsme dosáhli v případě koncentrací glukózy v CSF., Objective: 1. Evaluate the numbers of cells in the cerebrospinal fluid (CSF), glucose concentrations in the CSF, values of the glucose quotient (Q glu. ), lactate concentrations in the CSF and values of the coefficient of energy balance (KEB) as indicators of intensity of the inflammatory process in the CSF in groups of patients without CNS impairment, with slight serous inflam - mation of non-infectious aetiology in the CNS, with serous inflammation of infectious aetiology in the CNS and of patients with purulent inflammation in the CNS with extracellular bacteria in pathogenesis. 2. Compare the information potential of the used parameters of the glucose energy metabolism in the CSF compartments in our group of the investigated patients, i.e. concentrations of glucose in the CSF, values of the Q glu. , concentrations of lactate in the CSF and values of the KEB. Design: Retrospective study. Material and Methods: We examined 133 CSF specimens in patients without CNS impairment, 227 CSF specimens in pa - tients with slight serous inflammation with intrathecal synthesis of immunoglobulins of non-infectious aetiology in the CNS, 208 CSF specimens in patients with serous inflammation of infectious aetiology in the CNS and 140 CSF specimens in patients with purulent inflammation in the CNS with extracellular bacteria in pathogenesis. The objects of our interest were numbers of cells in the CSF, concentrations of glucose in the CSF, values of the Q glu. , concentrations of lactate in the CSF and values of the KEB. The D’Agostino Omnibus test, the Kruskal-Wallis test with follow-up post hoc analysis using the Dunn’s method, the Spearman correlation coefficient and the multinomial logistic regression analysis were used for statis - tical analysis of the examined parameters. Results: We did not find any changes in the numbers of cells in the CSF and in energy ratios in the CSF compartment of patients without CNS impairment. We found raised numbers of cells in the CSF and slight alterations of the glucose quo - tients, lactate concentrations in the CSF and the values of the KEB only in some patients with slight serous inflammations of non-infectious aetiology in the CNS. We observed manifestations of conspicuously increased intensity of inflammation in the numbers of cells in the CSF, lactate concentrations in the CSF and the values of the KEB in patients with serous inflam - mations of infectious aetiology in the CNS. Very high intensity of purulent inflammation in the CNS of bacterial aetiology was well apparent in all the evaluated parameters. Concerning the relationship, either direct or indirect, between the number of cells in the CSF and the other parameters, we found the highest correlation between the number of cells in the CSF and the values of the KEB ( ρ = -0.770), followed by the lactate concentrations in the CSF ( ρ = 0.734), the Q glu. ( ρ = -0.676) and the glucose concentrations in the CSF ( ρ = -0.544). We verified the applicability of the parameters mentioned above for prediction of the intensity of inflammation in the CNS via multinomial logistic regression analysis. The number of cells and the KEB, with 71.9 % and 71.6 % respectively, has the highest prediction potential of the correctly classified patients. They were followed by the lactate concentration in the CSF with 64.7 %, the Q glu. with 58.8 % and the glucose concentration with 54.7 % of the correctly classified patients. Conclusion: Our study supports the applicability of the numbers of cells in the CSF, the glucose concentrations in the CSF, the values of the Q glu. , the lactate concentrations in the CSF and the values of the KEB for diagnosing CSF impairment and for monitoring the intensity of inflammation in the CNS. Further, the results enabled determination of the information potential of the energy parameters. The values of the KEB were most suitable for evaluation of the intensity of inflammation in the CNS. Less suitable results were achieved in case of the lactate concentrations in the CSF. Even worse results were observed in case of the values of Q glu. and the least suitable results were observed in case of the glucose concentrations in the CSF., Kelbich P., Hejčl A., Procházka J., Selke Krulichová I., Peruthová J., Hanuljaková E., Špička J., and Literatura
Type I diabetes mellitus (DM1) is a complex disease with adverse effects on organs and tissues despite compensation by insulin treatment. The goal of our study was to study how kidney diseases change 31P MR parameters of muscle metabolism in DM1 patients with respect to gender. 51 DM1 patients (19 m/14 f without and 13 m/5 f with nephropathy) and 26 (14 m/12 f) healthy volunteers were examined using 31P magnetic resonance spectroscopy at 3T tomograph at rest, and during and after a calf muscle exercise. The exercise consisted of a six-minute plantar flexion using a pedal ergometer followed by a six-minute recovery. It is reflected by reduced relative β-ATP and increased Pi and phosphodiester signals to phosphocreatine (PCr) at rest and prolongation of the PCr recovery time after the exercise. Measurement on healthy volunteers indicated differences between males and females in pH at the rest and after the exercise only. These differences between patients groups were not significant. We have proven that nephropathy affects the metabolism in diabetic patients and our results confirm significant difference between patients with and without nephropathy. Gender differences in pH were observed only between male and female healthy volunteers., P. Sedivy, M. Dezortova, M. Drobny, Z. Vlasakova, V. Herynek, M. Hajek., and Obsahuje bibliografii
Huntington’s disease (HD) is a demential, neurodegenerative inheritable disease affecting middle-aged patients. HD is characterized by uncontrolled choreiform movements, psychiatric symptoms and cognitive decline. Histopathological changes in HD brains reveal a considerable damage to basal ganglia, particularly affecting middle-sized spiny neurons from the caudate-putamen region. Neurochemical changes are specifically oriented to deplete GABAergic and cholinergic systems, while molecular alterations include an increased expression of CAG trinucleotide at exon 1 from the huntingtin (htt) gene, as well as aggregation of mutant htt. Although several hypotheses regarding the mechanisms by which neurotoxicity is triggered in HD brains have been suggested on the basis of experimental evidence, so far it remains not clear which of them are predominant or whether they are complementary. Recent experimental evidence through transgenic mice models reveal an interesting inter action between expanded CAG triplets, mutant htt, and the increase in toxic metabolites from the kynurenine pathway. Further evidence supports the assumption that different toxic mechanisms (i.e. excitotoxicity, energy metabolism impairment, inflammatory events, oxidative stress, etc.) are confluent and depend on each other. In this review we will briefly summarize some of those findings and propose a final integrative hypothesis for HD., V. Pérez-de la Cruz, A. Santamaría., and Obsahuje bibliografii a bibliografické odkazy
Cíl studie: Výše termického efektu (TE) parenterální výživy (PV) podané těžce nemocným není známá. Cílem naší práce bylo změřit TE a dynamiku oxidace energetických substrátů PV v této populaci. Typ studie: Prospektivní. Název a sídlo pracoviště: Klinika gerontologická a metabolická, Fakultní nemocnice Hradec Králové. Materiál a metody: Do studie bylo zařazeno 21 pacientů interní JIP (APACHE II skóre 15,1 ± 7,9). Po 12hodinovém lačnění (infuze krystaloidů) jim byla po dobu 24 hodin podávána standardní PV (aminokyseliny – 1,2; tuk LCT/MCT – 0,5; glukóza – 4 mg/kg/min). Ve srovnání s běžnou praxí se jednalo o výživu relativně bohatou na sacharidy s nízkou dávkou tuku. Energetický výdej (EV) a oxidace energetických substrátů byla měřena metodou indirektní kalorimetrie před zahájením PV a následně po 2, 5, 12 a 24 hodinách jejího podávání. Byl posouzen vliv tíže onemocnění (APACHE II skóre) na sledované parametry. Výsledky: TE parenterální výživy dosáhl maxima 6,6 ± 9,5 % (p = 0,004) po 5 hodinách od zahájení infuze PV, poté postupně klesal k výchozí hodnotě. Velikost TE nebyla ovlivněna tíží základního onemocnění. Oxidace tuku byla při PV suprimována z počátečních 0,88 ± 0,60 na minimum 0,02 ± 1,0 mg/kg/min (p < 0,001) ve 12. hodině. Míra suprese pozitivně korelovala s tíží onemocnění (R = 0,56; p = 0,018). Oxidace sacharidů rostla během parenterální výživy z úvodních 1,19 ± 1,67 na maximum 3,46 ± 2,14 mg/kg/min (p < 0,001) ve 12. hodině. Míra zvýšení oxidace sacharidů při PV pozitivně korelovala s tíží onemocnění (R = 0,52; p = 0,03). Závěr: Termický efekt PV podané těžce nemocným je přechodný jev a téměř vymizí po 12 hodinách nutriční léčby. Není ovlivněn tíží stavu. Míra suprese oxidace tuků i nárůst oxidace sacharidů během PV jsou přímo úměrné tíží základního onemocnění., Objective: The thermic effect (TE) of parenteral nutrition (PV) is not known in severely ill patients. The aim of our study was to measure TE and the dynamics of substrates oxidation during PV in this population. Design: Prospective. Settings: Department of Gerontology and Metabolic Care, University Hospital in Hradec Králové. Material and methods: After a fastening period of 12 hours (infusion of saline) we administered to ICU patients (n = 21) total PV of the following composition: amino acids 1.2 mg, fat 0.5 mg, glucose 4.0 mg per minute per kilogram of ideal body weight. Prior to PV administration and subsequently 2, 5, 12, and 24 hour later energy expenditure (EV) and oxidation of energy substrates were measured by indirect calorimetry. The correlation between assessed parameters and APACHE II score was searched. Results: TE reached its maximum 5 hours after the onset of PV (6.6 ± 9.5% (p = 0.004)) and then it gradually declined. It was not infl uenced by disease severity. Lipid oxidation was suppressed from the fasting level of 0.88 ± 0.60 to the minimum of 0.02 ± 1.0 mg/kg/min (p < 0.001), whereas glucose oxidation increased from 1.19 ± 1.67 to the maximum of 3.46 ± 2.14 mg/kg/min (p < 0.001) in the 12th hour. The rate of suppression of lipid oxidation by PV was directly correlated with APACHE II score (R = 0.56; p = 0.018) Conclusions: The TE of PV administered to medical ICU patients was a transient phenomenon. Its average magnitude was 6.6%. The rate of suppression of lipid oxidation as well as increase in carbohydrates oxidation during PV were directly correlated to the severity of underlying illness., Bajnárek J., Maňák J., Vyroubal P., Hyšpler R., Sobotka L., and Lit.: 31
Empagliflozin je zástupcem nové skupiny antidiabetik, které způsobují glykosurii blokováním zpětné resorpce glukózy v proximálním tubulu ledviny. Ztráta 50–100 g glukózy/24 hod močí vede ke snížení glykemie na lačno, a především pak glykemie postprandiální, ztrátě energie 200–400 kcal/den a snížení krevního tlaku. Účinnost léčby neklesá v čase, protože není závislá na vlastní produkci inzulinu. Práce je zaměřena na hodnocení bezpečnosti moderní léčby empagliflozinem, který se v nejbližší době objeví v portfoliu antidiabetik v ČR. Rozborem studií, které byly provedeny za účelem hodnocení bezpečnosti a účinnosti léčby empagliflozinem se ukazuje, že se jedná o velmi účinný, ale zároveň bezpečný a dobře snášený lék. Při léčbě je popisován vyšší výskyt genitálních infekcí, zvláště u žen, které mají většinou lehký průběh. Lék nezpůsobuje hypoglykemie. V kombinaci se sulfonylureou však může k hypoglykemiím docházet. Empagliflozin u osob okolo 60 let věku nevyvolává klinicky významnou dehydrataci ani hypotenzi, nicméně určité opatrnosti s léčbou empagliflozinem je třeba u velmi starých, křehkých pacientů. Velkou výhodou empagliflozinu jsou klinicky nevýznamné interakce s léky a velmi jednoduché dávkování 1 tableta/den per os. Závěrem lze konstatovat, že je empagliflozin velmi účinné perorální antidiabetikum s možnosti širokého použití prakticky ve všech stadiích diabetu 2. typu v monoterapii nebo kombinaci s jakoukoliv zavedenou medikací. Léčba je spojena s poklesem hmotnosti a poklesem krevního tlaku. Lék je účinný a bezpečný až do eGF 0,75 ml/s, při nižší hodnotě je třeba léčbu empagliflozinem ukončit. Výskyt vedlejších účinků je minimální s výjimkou zvýšeného výskytu genitálních infekcí především u žen a zvýšeného rizika hypoglykemie, pokud je empagliflozin kombinován se sulfonylureou., Empagliflozin is agent of new antidiabetic drugs that cause glycosuria blocking the glucose reuptake in the proximal tubule. The loss of 50-100 g of glucose / 24 hours in the urine results in a reduction of fasting glucose, especially post-prandial glucose, the energy expenditure of 200-400 kcal / day and blood pressure lowering. Treatment efficacy does not decrease over time, as it is not dependent on its own insulin production. The work evaluates the safety of modern treatment with empagliflozin which will soon appear in the portfolio of antidiabetic agents in the Czech Republic. The conducted studies with a special focus on empagliflozin treatment have shown high efficacy, safety and good tolerability of drug. It has been described a higher incidence of genital infections with non-severe course, especially in women. The drug does not cause hypoglycaemia. In combination with sulfonylurea hypoglycaemia may occur. Empagliflozin does not cause clinically significant dehydration or hypotension in patients about 60 years of age, but some caution in empagliflozin treatment should be in elderly and fragile patients. The big convenience of empagliflozin is its clinically non-significant interactions with other drugs and simple dosage of 1 tablet / day orally. In conclusion, empagliflozin is highly effective oral antidiabetic agent with a potential of wide application in all stages of type 2 diabetes in monotherapy or combined with other medication. The treatment is associated with weight loss and blood pressure lowering. The drug is effective and safe until eGFR 45 ml / s, in lower values the treatment should be discontinued. The occurrence of side effects is rare, except increased incidence of genital infections especially in women and increased risk of hypoglycaemia when empagliflozin is combined with sulfonylurea., and Zdeněk Rušavý
a1_The purpose of the present study was to compare the ontogenetic development of the activity of myocardial energy-supplying enzymes in two mammalian species, differing significantly in their level of maturation at birth. The animals were investigated during the late prenatal period and 2, 7, 14, 21, 25, 30, 63, 120 and 730 days after birth in the rat and 2, 21, 84 and 175 days in the guinea-pig. The following enzymes were assayed in the right and left ventricular myocardium: lactate dehydrogenase (LDH, lactate uptake and/or formation), triose phosphate dehydrogenase (TPDH, carbohydrate metabolism), glycerol phosphate dehydrogenase (GPDH, glycerol-P shuttle)), hexokinase (HK, glucose phosphorylation), malate dehydrogenase (MDH, tricarboxylic cycle), citrate synthase (CS, tricarboxylic cycle) and hydroxyacyl-CoA dehydrogenase (HOADH, fatty acid breakdown). The rat heart, highly immature at birth, exhibits three different developmental patterns of energy-supplying enzymes, identical in both ventricles: (i) two mitochondrial enzymes of aerobic metabolism (CS, HOADH) and GPDH have a relatively low activity at the end of prenatal life; thereafter their activity steadily increases, approaching the adult levels between the 3rd and 4th postnatal weeks. A significant decrease was observed between the 4th and 24th months. (ii) MDH and LDH: prenatal values were significantly higher as compared with the 2nd postnatal day; after this period the activities increased up to adulthood (4 months) and decreased during senescence. (iii) The activities of HK and TPDH are characterized by only moderate changes during development. HK differs from all other enzymes by the highest prenatal values, which exceed even adult values. In contradiction to the rat heart, the developmental differences in more mature guinea-pig heart were significantly less pronounced., a2_The only ontogenetic differences observed were the lower activities of enzymes connected with aerobic metabolism at the end of the prenatal period. Our results point to possible differences in the development of adaptive metabolic pathways in animals with different levels of maturation at birth., A. Bass, M. Stejskalová, A. Stieglerová, B. Ošťádal, M. Šamánek., and Obsahuje bibliografii
The activities of cytochrome c oxidase and FoF1-ATPase as well as the content of cytochromes cc1, aa3, and b were investigated in free brain mitochondria in the course of postnatal development and aging. The results show an increase of Vmax of both enzymes during postnatal development (between day 5 and 30). During the following phase ending at the age of 6 months, a decrease of FoF1-ATPase and cytochrome c oxidase activity occurs. From 6 to 12 months of age the activity of these enzymes did not change. The KM for both enzymes remained unchanged during the whole period observed. The content of cytochromes increased from the low values found in young rats, reached the highest values at around one month, and decreased till the age of 3 months. Later, their content in brain mitochondria did not markedly change. Our results suggest that the metabolic maturation of brain mitochondria differs in several aspects from the same process in other tissues, mainly in the time course. This is probably due to the unique role of neural tissue in the organism., M. Kalous, H. Rauchová, Z. Drahota., and Obsahuje bibliografii
This study investigated the effects of riboflavin on energy metabolism in hypoxic mice. Kunming mice were fed diets containing riboflavin at doses of 6, 12, 24 and 48 mg/kg, respectively for 2 weeks before exposure to a simulated altitude of 6000 m for 8 h. Changes of riboflavin status and energy metabolism were assessed biochemically. Simultaneously, a 1H nuclear magnetic resonance (NMR) based metabolomic technique was used to track the changes of plasma metabolic profiling. It was found that the content of hepatic riboflavin was decreased and erythrocyte glutathione activation coefficient was elevated significantly under hypoxic condition. Meanwhile, increased plasma pyruvate, lactate, β-hydroxybutyrate and urea, as well as decreased plasma carnitine were observed. Riboflavin supplementation improved riboflavin status remarkably in hypoxic mice and decreased plasma levels of pyruvate, free fatty acids and β-hydroxybutyrate significantly. Plasma carnitine was increased in response to riboflavin supplementation. Results obtained from 1H NMR analysis were basically in line with the data from biochemical assays and remarkable changes in plasma taurine, choline and some other metabolites were also indicated. It was concluded that riboflavin requirement was increased under acute hypoxic condition and riboflavin supplementation was effective in improving energy metabolism in hypoxic mice., Y. P. Wang, J. Y. Wei, J. J. Yang, W. N. Gao, J. Q. Wu, C. J. Guo., and Obsahuje bibliografii