catechol
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Soubor 65 pacientů (22 žen, 43 mužů) s Parkinsonovou nemocí (PN) v pokročilém stadiu byl v období srpen I999 - leden 2001 léčen novým inhibitorem katechol-0-metyl-transferázy (COMT inhibitorem), entakaponem. Přípravek Comtan® (entakapon) byl podán pacientům s PN ke stávající L-DOPA a jiné antiparkinsonské terapii k léčbě fluktuací klinickému stavu. Průměrný věk souboru v době zahájení léčby entakaponem byl 66,6± 8,4 roku, průměrná doba trvání onemocnění 9,9±5,3 roku, průměrný věk v době prvních klinických příznaků onemocnění 56,7±10,3 roku, průměrný stupeň onemocnění podle Hoehnové a Yahra před zahájením léčby entakaponem byl 3,1±0,7, průměrná doba trvání fluktuací klinického stavu 5,4±3,7 roku, průměrná doba podávání L-DOPA 8,3±4,6 roku. Před zahájením terapie entakaponem bylo u každého pacienta stanoveno skóre UPDRSIII (motorické vyšetření), skóre UPDRS TV (komplikace terapie), průměrná denní doba bdělého dne strávená v off-stavu hybnosti (stavu špatné hybnosti) a on-stavu hybnosti (stavu dobré hybnosti). Dále byl proveden odběr krevního vzorku k laboratorní analýze jatemích transamináz. Průměrná denní dávka L-DOPA před zahájením terapie byla 1091+474 mg. Nejkratší doba sledování k hodnocení účinku terapie entakaponem byla 4 měsíce, nejdelší 17 měsíců (průměrná doba sledování souboru byla 12,1±3,9 měsíců). 12 pacientů (18,5 %), u nichž byla zahájena léčba entakaponem, mělo v anamnéze předchozí nesnášenlivost nebo komplikace při terapii dopaminergními agonisty, u dalších 10 pacientů (15,4 %) byly přítomny psychické komplikace onemocnění (kognitivní deficit, halucinace), které limitovaly nasazení jiné antiparkinsonské terapie kromě L-DOPA. Při následujících kontrolách po zahájení léčby entakaponem byly k hodnoceni účinku terapie použity škály UPDRS III (průměrný pokles o 27,8 %), UPDRS IV (průměrný pokles o 31,9 %), byl hodnocen čas strávený v off-stavu hybnosti (snížení o 2,3±1,4 hodiny), dále čas strávený v on-stavu hybnosti (zvýšení o 1,7±1,2 hodiny) - vše statisticky signifikantní na hladině významnosti p < 0,0001 oproti výchozím hodnotám před zahájením terapie entakaponem. Průměrná dávka entakaponu byla 812±197 mg; celková denní dávka L-DOPA 933±380 mg (pokles o 14,5 %, statistický signifikantní na hladině významnosti p < 0,05). Vedlejší účinky (dyspepsie, průjmy, ortostatická hypotenze, halucinace) se objevily celkem u 5 pacientů (7,7 %). Dyskineze byly zA^azněny u 11 pacientů (16,9 %), odezněly po redukci celkové denní dávky L-DOPA. Entakapon byl vysazen u 7 pacientů (10,8 %), většinou pro nedostatečný účinek (3 pacienti), non compliance (2 pacienti) nebo vedlejší účinky (2 pacienti). 2 pacienti během sledovaného období zemřeli na jiná interkurentní onemocnění (srdeční selhání, cévní mozková příhoda). U žádného z pacientů nebyla pozorována alterace jatemích transamináz ve srovnání s hodnotami před zahájením léčby entakaponem. Entakapon je účinný a bezpečný lék, vhodný k terapii fluktuací klinického stavu u pacientů v pokročilém stadiu Parkinsonovy nemoci. Entakapon signifikantně prodlužuje čas bdělého dne strávený v on-stavu hybnosti, snižuje procento času v off-stavu hybnosti. Entakapon nepůsobí alteraci jatemích transamináz. Lze jej s úspěchem užít i u pacientů s psychickými komplikacemi onemocnění nebo předchozí anamnézou nesnášenlivosti léčby dopaminergními agonisty.
A group of 65 patients (22 women. 43 men) with Parkinson's disease (PD) in the advanced stage was treated between August 1999 and January 2001 by a new catechol-0-methyl-transferase inhibitor (COMT inhibitor), entacapone. The preparation Comtan®(entacapone) was administered to patients with PD in addition to L-DOPA and other antiparkinsonian therapy to treat fluctuations of the clinical status. The mean age of the group at the time when entacapone therapy was started was 66.6±8.4 years. the mean duration of the disease 9.9±5.3 years, the mean age at the time of the first clinical symptoms was 56.7+10.3 years, the mean grade of the disease according to Hoehn and Yahr before initiation of entacapone treatment was 3.1 ±0.7, the mean period of fluctuations of the clinical status was 5.4±3.7 years, the mean period of L-DOPA administration 8.3±4.6 years. Before entacapone treatment was started in every patients the UPDRS III score (motor examination), the UPRDSIV score (complications of treatment), the mean daily period of the wakeful day spent in the off-state (state of poor mobility) and on-state of mobility (state of adequate mobility) were assessed. Furthermore a blood sample was taken for laboratory analysis of liver transaminases. The mean daily dose of L-DOPA before the beginning of therapy was 1091+474 mg. The shortest follow up period for evaluation of the effect of entacapone therapy was 4 months, the longest 17 months (mean period of follow up of the group was 12.1±3.9 months). Twelve patients (18.5 %) where entacapone treatment was started had a history of previous intolerance or complications during treatment with dopaminergic agonists, in another 10 patients (15.4 %) mental complications were present (cognitive deficiency, hallucinations), which limited the administration of other antiparkinsonian therapy except L-DOPA. During subsequent check-up examinations after initiation of entacapone treatment, for evaluation of the effects of therapy scales UPDRS III (mean decline by 27.8 %), UPDRS IV (mean decline by 31.9 %) were used and the time spent in the off-state of mobility (reduction by 2.3±1.4 hours, the time spent in the on-state of mobility (increase by 1.7+1.2 hours) were assessed - all statistically significant at the level of p < 0.0001 as compared with baseline values before the initiation of entacapone treatment. The mean entacapone dose was 812+197 mg. The total daily dose of L-DOPA 933±380 mg (decline by 14.5 % statistically significant at the level of p < 0.05). Side-effects (dyspepsia, diarrhoea, orthostatic hypotension, hallucinations) were recorded in 5 patients (7.7 %). Dyskinesias became more marked in 11 patients (16.9 %), they receded after reduction of the total daily L-DOPA dose. Entacapone was discontinued in 7 patients (10.8 %), usually because of an inadequate effect (3 patients), non-compliance (2 patients) or side-effects y V lents) Two patients died during the follow up period from other intercurrent diseases (heart failure, cerebrovascular attack). In none of the patients alteration of liver transaminases was observed as compared with values before entacapone treatment. Entacapone is an effective and safe preparation suitable for treatment of fluctuations of the clinical status of patients with the advanced stage of Parkinson's disease. Entacapone prolongs significantly the time of the wakeful day spent in the on-state of mobility, it reduces the percentage of time in the off-state of mobility. Entacapone does not cause a change of liver transaminases. It can be successfully used also in patients with mental complications of the disease or those with a previous history of intolerance of dopaminergic agonists.
The purpose of the present study was to purify and characterize the catechol 1,2-dioxygenase (EC 1.13.11.1; catechol-oxygen 1,2-oxidoreductase; C12O) enzyme from the local isolate of Pseudomonas putida. This enzyme catalyzes the initial reaction in the ortho-pathway for phenol degradation in various gram-negative bacteria, including the genus of Pseudomonas. Pseudomonads are commonly used in the biodegradation of xenobiotics due to their versatility in degrading a wide range of chemical compounds. Eighty-nine soil samples were taken from the contaminated soil of the Midland Refineries Company (MRC) of Al-Daura refinery area at Baghdad from April to August 2021. The samples were grown in a mineral salt medium containing 250 mg per L of phenol to test their ability to biodegrade phenol. The pH was adjusted to 8.0 at 30 °C using a shaking incubator for 24-48 h. A number of 62 (69.6%) isolates of the total number were able to degrade phenol efficiently. The findings of the VITEK system and the housekeeping gene 16S rDNA confirmed that out of the positive isolates for phenol degradation, 36 from 62 (58.06%) were identified as Pseudomonas spp. isolates. Those isolates were distributed as P. aeruginosa 30 (83.3%) and P. putida 6 (16.6%). The enzyme production capabilities of the isolates were evaluated, and the highest activity was 2.39 U per mg for the isolate No. 15 which it was identified as P. putida. The previous isolate was selected for enzyme production, purification, and characterization. The enzyme was purified using ion exchange and gel filtration chromatography, with a combined yield of 36.12% and purification fold of 15.42 folds. Using a gel filtration column, the enzyme's molar mass was calculated to be 69 kDa after purification. The purified enzyme was stable at 35 °C and a pH of 6.0.
- MeSH
- bakteriální proteiny metabolismus genetika chemie izolace a purifikace MeSH
- biodegradace * MeSH
- fenol * metabolismus MeSH
- fylogeneze MeSH
- katechol-1,2-dioxygenasa * metabolismus genetika MeSH
- koncentrace vodíkových iontů MeSH
- Pseudomonas putida * enzymologie genetika metabolismus MeSH
- půdní mikrobiologie * MeSH
- RNA ribozomální 16S genetika MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
OBJECTIVES: Candida tropicalis yeast is a microorganism that possesses high tolerance for phenol and shows strong phenol degrading activity. This yeast is capable of utilizing phenol as the sole carbon and energy source. While the enzyme participating on the first step of phenol biodegradation, NADPH-dependent phenol hydroxylase, has already been characterized, information on the enzyme participating in the second step of its degradation, catechol 1,2-dioxygenase, is scarce. The development of the procedure suitable for catechol 1,2-dioxygenase isolation and partial characterization of this enzyme are the aims of this study. METHODS: Combination of chromatography on DEAE-Sepharose and gel-permeation chromatography on Sephadex G-100 was used for isolation of cytosolic catechol 1,2-dioxygenase from C. tropicalis yeast. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel chromatography on Sephadex G-100 were used to evaluate the molecular mass of the enzyme. The enzyme activity was followed by HPLC (catechol consumption and/or cis,cis-muconic acid formation). RESULTS: Using the isolation procedure consisting of chromatography and re-chromatography on a column of DEAE-Sepharose and gel filtration on Sephadex G-100, catechol 1,2-dioxygenase was purified from C. tropicalis cytosol to homogeneity. Catechol 1,2-dioxygenase was found to be a homodimer with a subunit molecular mass of 30000 +/- 5000. The enzyme oxidized catechol producing cis,cis-muconic acid. The optimal temperature and pH were 30 degrees C and 7.7, respectively. CONCLUSIONS: The data are the first report showing the isolation of eukaryotic catechol 1,2-dioxygenase from C. tropicalis to homogeneity and its partial characterization.
- MeSH
- Candida tropicalis enzymologie metabolismus MeSH
- chromatografie MeSH
- cytosol chemie enzymologie metabolismus MeSH
- dextrany MeSH
- dimerizace MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- fenol chemie MeSH
- fungální proteiny chemie izolace a purifikace metabolismus MeSH
- gelová chromatografie MeSH
- katechol-1,2-dioxygenasa chemie izolace a purifikace metabolismus MeSH
- katecholy chemie metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- kyselina sorbová analogy a deriváty chemie metabolismus MeSH
- oxidace-redukce MeSH
- teplota MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Východisko: Jedinci závislí na metamfetaminu mají jedenáctinásobně vyšší riziko rozvoje psychotických příznaků oproti běžné populaci. Dopamin, jeden z klíčových neurotransmitérů v etiopatogenezi psychózy, je metabolizován katechol-O-methyltransferázou (COMT). Polymorfismus Val158Met genu pro COMT mění aktivitu tohoto enzymu až čtyřnásobně, a tím ovlivňuje hladinu dopaminu v mozku. Cílem studie bylo zjistit, zda jedinci závislí na metamfetaminu s výskytem psychotických příznaků se liší od osob závislých na metamfetaminu bez psychotických příznaků pokud jde o genotyp nebo výskyt jednotlivých alel polymorfismu COMT. Metody: Do studie byli zavzati pacienti závislí na metamfetaminu, kteří s výzkumem souhlasili. Psychotické příznaky v rámci Dg. F15.50 až F15.53 byly diagnostikovány pomocí klasifikace MKN-10. Genotyp COMT byl určen metodou PCR (polymerase chain reaction). Výsledky: Do studie vstoupilo celkem 83 osob (ženy N = 25) ve věkovém rozmezí 18 – 38 let (medián 22 let) závislých na metamfetaminu. Psychotické příznaky se vyskytly u 18 pacientů (ženy N = 8).Nebylo prokázáno, že výskyt psychotických příznaků u osob závislých na metamfetaminu by statisticky významně souvisel s genotypem COMT nebo jednotlivými alelami (P = N.S.; Fisherův přesný test). Závěr: Výsledek studie mohl být ovlivněn hodnocením celého souboru bez rozlišení pohlaví, skrytými etnickými faktory nebo interakcí vyšetřovaného polymorfismu s jinými geny či faktory prostředí.
Background: The risk of psychotic symptoms in methamphetamine dependent individuals is eleven times higher as compared to the general population. Dopamine, which is one of the key neurotransmitters in etiopathogenesis of psychosis, is metabolised by catechol-O-methyltransferase (COMT). The COMT gene Val158Met polymorphism changes activity of the COMT enzyme up to fourfold and thus influences the dopamine activity in the brain. The aim of the study was to ascertain whether subjects dependent on methamphetamine with psychotic symptoms significantly differ from methamphetamine dependent individuals without psychosis in the Val158Met COMT genotype or allele frequency. Methods: Methamphetamine dependent inpatients who had signed the informed consent were enrolled into the study.We assessed psychotic symptoms according to the ICD-10 classification of mental disorders (Dgs F15.50 to F15.53). The COMT genotype was detected by the PCR (polymerase chain reaction). Results: Eighty-three patients (25 women) at the age of 18–38 years (median 22 years) dependent on methamphetamine entered the study. We found psychotic symptoms in 18 subjects (8 women). No significant association between psychosis in methamphetamine dependence and the Val158Met COMT genotype or allele frequency was proved (P = N.S.; Fisher´s Exact Test). Conclusions: Our results may have been influenced by the joint assessment of males and females in one sample, hidden ethnic stratification or interaction of the Val158Met COMT gene polymorphism with other genes and environmental factors.
- MeSH
- dopamin metabolismus MeSH
- dospělí MeSH
- financování organizované MeSH
- hodnocení rizik MeSH
- interpretace statistických dat MeSH
- katechol-O-methyltransferasa genetika MeSH
- lidé MeSH
- methamfetamin škodlivé účinky toxicita MeSH
- polymorfismus genetický genetika účinky léků MeSH
- poruchy spojené s užíváním psychoaktivních látek genetika komplikace MeSH
- psychotické poruchy etiologie genetika MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
OBJECTIVES: About 25,000 serious methamphetamine abusers live in the Czech Republic among the total population of 10 million. Dependence on methamphetamine is markedly related to the brain neurotransmitter dopamine, metabolised by catechol-O-methyltransferase enzyme. The main aim of the study was to ascertain whether the Val158Met catechol-O-methyltransferase gene polymorphism is associated with methamphetamine dependence in this Central European country. Methods: One hundred and twenty-three subjects dependent on methamphetamine (women N=44), parents of sixty-seven dependent individuals, and four hundred healthy controls (women N=250) were involved into the study. We performed a population-based as well as family-based genetic association studies. Results: We did not find any significant association between the Val158Met catechol-O-methyltransferase gene polymorphism and methamphetamine dependence using the population-based or family-based design (p=0.41-0.66; Chi-Square Test or UNPHASED program, Version 3.1.4, respectively). We found a trend toward a statistically significant difference between the Val allele carriers and Met/Met homozygotes in the frequence of psychotic symptoms induced by methamphetamine (more frequent in Val carriers; p=0.062; Chi-Square Test). Conclusion: Further research involving haplotype analysis and other dopamine-related genetic polymorphisms in large populations is needed. More attention should also be paid to possible role of the Val158Met catechol-O-methyl-transferase gene polymorphism in individual clinical subtypes of dependence on methamphetamine involving e.g. psychotic features or violence.
- MeSH
- bodová mutace MeSH
- dospělí MeSH
- frekvence genu MeSH
- genetická predispozice k nemoci genetika MeSH
- haplotypy MeSH
- katechol-O-methyltransferasa genetika MeSH
- lidé MeSH
- methamfetamin škodlivé účinky MeSH
- mladiství MeSH
- mladý dospělý MeSH
- polymorfismus genetický MeSH
- poruchy spojené s užíváním amfetaminu genetika MeSH
- psychotické poruchy genetika MeSH
- stimulanty centrálního nervového systému škodlivé účinky MeSH
- zdraví rodiny MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Alzheimer's disease (AD) is one of the most significant neurodegenerative disorders and its symptoms mostly appear in aged people. Catechol-o-methyltransferase (COMT) is one of the known target enzymes responsible for AD. With the use of 23 known inhibitors of COMT, a query has been generated and validated by screening against the database of 1500 decoys to obtain the GH score and enrichment value. The crucial features of the known inhibitors were evaluated by the online ZINC Pharmer to identify new leads from a ZINC database. Five hundred hits were retrieved from ZINC Pharmer and by ADMET (absorption, distribution, metabolism, excretion, and toxicity) filtering by using FAF-Drug-3 and 36 molecules were considered for molecular docking. From the COMT inhibitors, opicapone, fenoldopam, and quercetin were selected, while ZINC63625100_413 ZINC39411941_412, ZINC63234426_254, ZINC63637968_451, and ZINC64019452_303 were chosen for the molecular dynamics simulation analysis having high binding affinity and structural recognition. This study identified the potential COMT inhibitors through pharmacophore-based inhibitor screening leading to a more complete understanding of molecular-level interactions.
- MeSH
- Alzheimerova nemoc farmakoterapie enzymologie patofyziologie MeSH
- exprese genu MeSH
- farmaceutické databáze MeSH
- inhibitory katechol-O-methyltransferasy chemie farmakologie MeSH
- interakční proteinové domény a motivy MeSH
- katechol-O-methyltransferasa chemie MeSH
- kinetika MeSH
- konformace proteinů, alfa-helix MeSH
- konformace proteinů, beta-řetězec MeSH
- lidé MeSH
- ligandy MeSH
- nootropní látky chemie farmakologie MeSH
- rychlé screeningové testy * MeSH
- simulace molekulární dynamiky MeSH
- simulace molekulového dockingu MeSH
- substrátová specifita MeSH
- terciární struktura proteinů MeSH
- termodynamika MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Acta dermato-venerologica ; vol. 44, suppl. 55
16 s. ; 26 cm
OBJECTIVE: Comamonas testosteroni Pb50 is a microorganism that possesses high tolerance for phenol and shows strong phenol degrading activity. This bacterial strain is capable of utilizing phenol as the sole carbon and energy source. Although examples are known in which the C. testosteroni utilizes phenol for growth or metabolism, much less information are known on the nature of the phenol-oxidizing enzymes in this microorganism. Therefore, the occurrence and cellular location of phenol hydroxylase (EC 1.14.13.7), the enzyme participating in the first step of phenol degradation, catalyzing its hydroxylation to catechol in a bacterial Comamonas testosteroni Pb50 strain grown in the presence of phenol as a sole carbon and energy source are the aims of this study. METHODS: Combination of fractionation with polyethylene glycol 6000 and gel permeation chromatography on columns of Sepharose 4B and Sephacryl S-300 was used for isolation of phenol hydroxylase detectable in the medium in which C. testosteroni was cultivated. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel chromatography on Sephacryl S-300 were used to evaluate the molecular mass of the enzyme. The enzyme activity was followed by HPLC (phenol consumption and/or catechol formation). RESULTS: Whereas low activity of phenol hydroxylase was detected in cytosol isolated from C. testosteroni, more than 16-fold higher activity of this enzyme was found in the medium in which C. testosteroni was cultivated. The presence of phenol hydroxylase extracellular activity suggests that this microorganism may secrete the enzyme into the extracellular medium. Using the procedure consisting of fractionation with polyethylene glycol 6000 and gel permeation chromatography on columns of Sepharose 4B and Sephacryl S-300, the enzyme was isolated from the medium to homogeneity. The formation of catechol mediated by purified phenol hydroxylase is strictly dependent on the presence of NADPH, which indicates that this enzyme is the NADPH-dependent phenol hydroxylase. The enzyme is a homotetramer having a molecular mass of 240 000, consisting of four subunits having a molecular mass of 60 000. The optimum pH of the enzyme for the phenol oxidation is pH 7.6. CONCLUSION: The results are the first report showing isolation and partial characterization of extracellular NADPH-dependent phenol hydroxylase of a bacterial C. testosteroni Pb50 strain capable of oxidizing phenol to catechol. The data demonstrate the progress in resolving the enzymes responsible for the first step of phenol degradation by bacteria.
- MeSH
- časové faktory MeSH
- Comamonas testosteroni enzymologie genetika MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- extracelulární prostor enzymologie genetika MeSH
- fenol metabolismus MeSH
- katalýza MeSH
- katecholy metabolismus MeSH
- klonování DNA MeSH
- koncentrace vodíkových iontů MeSH
- NADP metabolismus MeSH
- oxidace-redukce MeSH
- oxygenasy se smíšenou funkcí genetika izolace a purifikace metabolismus MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
