BACKGROUND: Recent years have witnessed a considerable increase in clinical trials of new investigational agents for Fabry disease (FD). Several trials investigating different agents are currently in progress; however, lack of standardisation results in challenges to interpretation and comparison. To facilitate the standardisation of investigational programs, we have developed a common framework for future clinical trials in FD. METHODS AND FINDINGS: A broad consensus regarding clinical outcomes and ways to measure them was obtained via the Delphi methodology. 35 FD clinical experts from 4 continents, representing 3389 FD patients, participated in 3 rounds of Delphi procedure. The aim was to reach a consensus regarding clinical trial design, best treatment comparator, clinical outcomes, measurement of those clinical outcomes and inclusion and exclusion criteria. Consensus results of this initiative included: the selection of the adaptative clinical trial as the ideal study design and agalsidase beta as ideal comparator treatment due to its longstanding use in FD. Renal and cardiac outcomes, such as glomerular filtration rate, proteinuria and left ventricular mass index, were prioritised, whereas neurological outcomes including cerebrovascular and white matter lesions were dismissed as a primary or secondary outcome measure. Besides, there was a consensus regarding the importance of patient-related outcomes such as general quality of life, pain, and gastrointestinal symptoms. Also, unity about lysoGb3 and Gb3 tissue deposits as useful surrogate markers of the disease was obtained. The group recognised that cardiac T1 mapping still has potential but requires further development before its widespread introduction in clinical trials. Finally, patients with end-stage renal disease or renal transplant should be excluded unless a particular group for them is created inside the clinical trial. CONCLUSION: This consensus will help to shape the future of clinical trials in FD. We note that the FDA has, coincidentally, recently published draft guidelines on clinical trials in FD and welcome this contribution.
- MeSH
- alfa-galaktosidasa genetika MeSH
- delfská metoda MeSH
- dospělí MeSH
- enzymová substituční terapie * MeSH
- Fabryho nemoc farmakoterapie genetika metabolismus patologie MeSH
- globosidy terapeutické užití MeSH
- glykolipidy terapeutické užití MeSH
- izoenzymy genetika MeSH
- klinické zkoušky jako téma * MeSH
- konsensus MeSH
- kvalita života MeSH
- ledviny účinky léků metabolismus patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- sfingolipidy terapeutické užití MeSH
- trihexosylceramidy terapeutické užití MeSH
- výsledek terapie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Trypanosoma brucei is an important human pathogen. In this study, we have focused on the characterization of FtsH protease, ATP-dependent membrane-bound mitochondrial enzyme important for regulation of protein abundance. We have determined localization and orientation of all six putative T.brucei FtsH homologs in the inner mitochondrial membrane by in silico analyses, by immunofluorescence, and with protease assay. The evolutionary origin of these homologs has been tested by comparative phylogenetic analysis. Surprisingly, some kinetoplastid FtsH proteins display inverted orientation in the mitochondrial membrane compared to related proteins of other examined eukaryotes. Moreover, our data strongly suggest that during evolution the orientation of FtsH protease in T. brucei varied due to both loss and acquisition of the transmembrane domain.
- MeSH
- Arabidopsis klasifikace enzymologie genetika MeSH
- Euglena gracilis klasifikace enzymologie genetika MeSH
- Euglena longa klasifikace enzymologie genetika MeSH
- exprese genu MeSH
- fylogeneze MeSH
- izoenzymy chemie genetika metabolismus MeSH
- konzervovaná sekvence MeSH
- Leishmania major klasifikace enzymologie genetika MeSH
- lidé MeSH
- mitochondriální membrány chemie enzymologie MeSH
- mitochondriální proteiny chemie genetika metabolismus MeSH
- mitochondrie enzymologie genetika MeSH
- molekulární evoluce * MeSH
- myši MeSH
- proteasy chemie genetika metabolismus MeSH
- proteinové domény MeSH
- protozoální proteiny chemie genetika metabolismus MeSH
- Saccharomyces cerevisiae klasifikace enzymologie genetika MeSH
- Trypanosoma brucei brucei klasifikace enzymologie genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Aberrant levels of histone modifications lead to chromatin malfunctioning and consequently to various developmental defects and human diseases. Therefore, the proteins bearing the ability to modify histones have been extensively studied and the molecular mechanisms of their action are now fairly well understood. However, little attention has been paid to naturally occurring alternative isoforms of chromatin modifying proteins and to their biological roles. In this review, we focus on mammalian KDM2A and KDM2B, the only two lysine demethylases whose genes have been described to produce also an alternative isoform lacking the N-terminal demethylase domain. These short KDM2A/B-SF isoforms arise through alternative promoter usage and seem to play important roles in development and disease. We hypothesise about the biological significance of these alternative isoforms, which might represent a more common evolutionarily conserved regulatory mechanism.
- MeSH
- izoenzymy nedostatek genetika metabolismus MeSH
- Jumonjiho doména s histondemethylasami nedostatek genetika metabolismus MeSH
- lidé MeSH
- nádory enzymologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Ten Enterobacteriaceae isolates collected in a Czech hospital carried blaKPC-positive plasmids of different sizes (∼30, ∼45, and ∼80 kb). Sequencing revealed three types of plasmids (A to C) with the Tn4401a transposon. Type A plasmids comprised an IncR backbone and a KPC-2-encoding multidrug resistance (MDR) region. Type B plasmids were derivatives of type A plasmids carrying an IncN3-like segment, while type C plasmids were IncP6 plasmids sharing the same KPC-2-encoding MDR region with type A and B plasmids.
- MeSH
- antibakteriální látky terapeutické užití MeSH
- beta-laktamasy genetika metabolismus MeSH
- Citrobacter freundii účinky léků enzymologie genetika izolace a purifikace MeSH
- enterobakteriální infekce farmakoterapie epidemiologie mikrobiologie MeSH
- Escherichia coli účinky léků enzymologie genetika izolace a purifikace MeSH
- exprese genu MeSH
- izoenzymy genetika metabolismus MeSH
- karbapenemy terapeutické užití MeSH
- Klebsiella pneumoniae účinky léků enzymologie genetika izolace a purifikace MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- mnohočetná bakteriální léková rezistence genetika MeSH
- Morganella morganii účinky léků enzymologie genetika izolace a purifikace MeSH
- nemocnice MeSH
- otevřené čtecí rámce MeSH
- plazmidy chemie klasifikace metabolismus MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- transpozibilní elementy DNA MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Lipoxygenases (LOX) constitute a heterogeneous family of lipid peroxidizing enzymes which catalyze the stereospecific insertion of molecular oxygen into polyunsaturated fatty acids. The primary products of LOX reaction are hydroperoxy fatty acids, which are rapidly reduced to the corresponding hydroxy derivatives. In mammalian organisms, there are 4 main LOX isoforms, 5-LOX, 8-LOX, 12-LOX, 15-LOX, constituting hydroperoxides with positional and stereo specificity. In the subsequent metabolic processes they provide significant cell signaling molecules and secondary messengers taking part in the development of inflammatory, tumorous and other diseases. Recent studies clarified the role of lipoxygenases in various diseases. The study of the structure and the active site of enzyme, preparation of recombinant forms with the targeted gene mutations have shown possibilities for better understanding the lipoxygenase role in various pathological processes. Searching for selective inhibitors of LOX isoenzymes is an active area of investigation. They can represent an important research tool or become a promising targeted therapy of a wide range of human diseases.
- MeSH
- izoenzymy genetika klasifikace MeSH
- kyselina arachidonová metabolismus MeSH
- lidé MeSH
- lipoxygenasy * farmakologie chemie klasifikace MeSH
- molekulární konformace MeSH
- mutace MeSH
- nenasycené mastné kyseliny metabolismus MeSH
- savci MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
The objective of this study was to perform molecular surveillance for assessing the spread of carbapenemase-producing Pseudomonas aeruginosa in Czech hospitals. One hundred thirty-six carbapenemase-producing isolates were recovered from 22 hospitals located throughout the country. Sequence type 357 (ST357) dominated (n = 120) among carbapenemase producers. One hundred seventeen isolates produced IMP-type (IMP-7 [n = 116] and IMP-1 [n = 1]) metallo-β-lactamases (MβLs), 15 produced the VIM-2 MβL, and the remaining isolates expressed the GES-5 enzyme. The blaIMP-like genes were located in three main integron types, with In-p110-like being the most prevalent (n = 115). The two other IMP-encoding integrons (In1392 and In1393) have not been described previously. blaVIM-2-carrying integrons included In59-like, In56, and a novel element (In1391). blaGES-5 was carried by In717. Sequencing data showed that In-p110-like was associated with a Tn4380-like transposon inserted in genomic island LESGI-3 in the P. aeruginosa chromosome. The other integrons were also integrated into the P. aeruginosa chromosome. These findings indicated the clonal spread of ST357 P. aeruginosa, carrying the IMP-7-encoding integron In-p110, in Czech hospitals. Additionally, the sporadic emergence of P. aeruginosa producing different carbapenemase types, associated with divergent or novel integrons, punctuated the ongoing evolution of these bacteria.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální chromozomy chemie MeSH
- beta-laktamasy genetika metabolismus MeSH
- epidemiologické monitorování MeSH
- exprese genu MeSH
- genomové ostrovy MeSH
- genotyp MeSH
- incidence MeSH
- integrony MeSH
- izoenzymy genetika metabolismus MeSH
- karbapenemy farmakologie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- mnohočetná bakteriální léková rezistence genetika MeSH
- multilokusová sekvenční typizace MeSH
- nemocnice MeSH
- pseudomonádové infekce epidemiologie mikrobiologie MeSH
- Pseudomonas aeruginosa účinky léků enzymologie genetika izolace a purifikace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- Geografické názvy
- Česká republika epidemiologie MeSH
The prevalence of obesity is rapidly increasing across the world. Physiologic alterations associated with obesity are known to alter enzyme expression and/or activities. As drug-metabolizing and antioxidant enzymes serve as defense system against potentially toxic compounds, their modulation might have serious consequences. In this work, we studied selected antioxidant and drug-metabolizing enzymes (DME) in monosodium glutamate-mouse model of obesity. Specific activities, protein, and mRNA expressions of these enzymes in liver as well as in small intestine were compared in obese male mice and in their lean counterparts. Furthermore, expression of the NF-E2-related factor 2 (Nrf2) and its relation to obesity were tested. Obtained results showed that obesity affects expression and/or activities of some DME and antioxidant enzymes. In obese mice, upregulation of UDP-glucuronosyltransferases 1A (UGT1A), NAD(P)H:quinone oxidoreductase 1 (NQO1), nuclear transcription factor Nrf2, and downregulation of some isoforms of glutathione S-transferases (GST) were observed. Most of these changes were tissue and/or isoform specific. NQO1 seems to be regulated transcriptionally via Nrf2, but other enzymes might be regulated post-transcriptionally and/or post-translationally. Enhanced expression of Nrf2 in livers of obese mice is expected to play a role in protective adaptation. In contrast, elevated activities of NQO1 and UGT1A may cause alterations in drug pharmacokinetics in obese individuals. Moreover, decreased capacity of GST in obese animals indicates potentially reduced antioxidant defense and weaker chemoprotection.
- MeSH
- faktor 2 související s NF-E2 genetika metabolismus MeSH
- glukuronosyltransferasa genetika metabolismus MeSH
- glutamát sodný MeSH
- glutathiontransferasa genetika metabolismus MeSH
- inbrední kmeny myší MeSH
- izoenzymy genetika metabolismus MeSH
- játra enzymologie metabolismus MeSH
- messenger RNA metabolismus MeSH
- modely nemocí na zvířatech * MeSH
- NAD(P)H dehydrogenasa (chinon) genetika metabolismus MeSH
- novorozená zvířata MeSH
- obezita enzymologie metabolismus MeSH
- orgánová specificita MeSH
- regulace genové exprese enzymů * MeSH
- střevní sliznice enzymologie metabolismus MeSH
- tenké střevo enzymologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The virulence of the Candida pathogens is enhanced by the production of secreted aspartic proteases, which therefore represent possible targets for drug design. Here, the crystal structure of the secreted aspartic protease Sapp2p from Candida parapsilosis was determined. Sapp2p was isolated from its natural source and crystallized in complex with pepstatin A, a classical aspartic protease inhibitor. The atomic resolution of 0.83 Å allowed the protonation states of the active-site residues to be inferred. A detailed comparison of the structure of Sapp2p with the structure of Sapp1p, the most abundant C. parapsilosis secreted aspartic protease, was performed. The analysis, which included advanced quantum-chemical interaction-energy calculations, uncovered molecular details that allowed the experimentally observed equipotent inhibition of both isoenzymes by pepstatin A to be rationalized.
- MeSH
- aspartátové proteasy chemie genetika izolace a purifikace metabolismus MeSH
- Candida chemie enzymologie genetika MeSH
- exprese genu MeSH
- fungální proteiny chemie genetika izolace a purifikace metabolismus MeSH
- inhibitory proteas chemie MeSH
- izoenzymy chemie genetika izolace a purifikace metabolismus MeSH
- katalytická doména MeSH
- kinetika MeSH
- krystalografie rentgenová MeSH
- kvantová teorie MeSH
- molekulární modely MeSH
- molekulární sekvence - údaje MeSH
- pepstatiny chemie MeSH
- sekundární struktura proteinů MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- strukturní homologie proteinů MeSH
- substrátová specifita MeSH
- termodynamika MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Aldehyde dehydrogenases (ALDHs) are responsible for oxidation of biogenic aldehyde intermediates as well as for cell detoxification of aldehydes generated during lipid peroxidation. So far, 13 ALDH families have been described in plants. In the present study, we provide a detailed biochemical characterization of plant ALDH2 and ALDH7 families by analysing maize and pea ALDH7 (ZmALDH7 and PsALDH7) and four maize cytosolic ALDH(cALDH)2 isoforms RF2C, RF2D, RF2E and RF2F [the first maize ALDH2 was discovered as a fertility restorer (RF2A)]. We report the crystal structures of ZmALDH7, RF2C and RF2F at high resolution. The ZmALDH7 structure shows that the three conserved residues Glu(120), Arg(300) and Thr(302) in the ALDH7 family are located in the substrate-binding site and are specific to this family. Our kinetic analysis demonstrates that α-aminoadipic semialdehyde, a lysine catabolism intermediate, is the preferred substrate for plant ALDH7. In contrast, aromatic aldehydes including benzaldehyde, anisaldehyde, cinnamaldehyde, coniferaldehyde and sinapaldehyde are the best substrates for cALDH2. In line with these results, the crystal structures of RF2C and RF2F reveal that their substrate-binding sites are similar and are formed by an aromatic cluster mainly composed of phenylalanine residues and several nonpolar residues. Gene expression studies indicate that the RF2C gene, which is strongly expressed in all organs, appears essential, suggesting that the crucial role of the enzyme would certainly be linked to the cell wall formation using aldehydes from phenylpropanoid pathway as substrates. Finally, plant ALDH7 may significantly contribute to osmoprotection because it oxidizes several aminoaldehydes leading to products known as osmolytes.
- MeSH
- aldehyddehydrogenasa chemie genetika metabolismus MeSH
- fylogeneze MeSH
- hrách setý enzymologie genetika MeSH
- izoenzymy chemie genetika metabolismus MeSH
- katalytická doména genetika MeSH
- kinetika MeSH
- krystalografie rentgenová MeSH
- kukuřice setá enzymologie genetika MeSH
- modely genetické MeSH
- molekulární modely MeSH
- molekulární sekvence - údaje MeSH
- NAD metabolismus MeSH
- rostlinné proteiny chemie genetika metabolismus MeSH
- rostliny enzymologie genetika MeSH
- sekvence aminokyselin MeSH
- stanovení celkové genové exprese MeSH
- substrátová specifita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
VIM-39, a VIM-1-like metallo-β-lactamase variant (VIM-1 Thr33Ala His224Leu) was identified in a clinical isolate of Klebsiella pneumoniae belonging to sequence type 147. VIM-39 hydrolyzed ampicillin, cephalothin, and imipenem more efficiently than did VIM-1 and VIM-26 (a VIM-1 variant with the His224Leu substitution) because of higher turnover rates.
- MeSH
- ampicilin metabolismus farmakologie MeSH
- antibakteriální látky metabolismus farmakologie MeSH
- beta-laktamasy genetika metabolismus MeSH
- beta-laktamová rezistence genetika MeSH
- biotransformace MeSH
- cefalothin metabolismus farmakologie MeSH
- exprese genu MeSH
- hydrolýza MeSH
- imipenem metabolismus farmakologie MeSH
- infekce bakteriemi rodu Klebsiella farmakoterapie mikrobiologie MeSH
- izoenzymy genetika metabolismus MeSH
- kinetika MeSH
- Klebsiella pneumoniae účinky léků enzymologie genetika izolace a purifikace MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- molekulární sekvence - údaje MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- techniky typizace bakterií MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Řecko MeSH
