The metabolic degradation of aldehydes is catalyzed by oxidoreductases from which aldehyde dehydrogenases (EC 1.2.1) comprise nonspecific or substrate-specific enzymes. The latter subset is represented, e.g., by NAD(+)-dependent aminoaldehyde dehydrogenases (AMADHs; EC 1.2.1.19) oxidizing a group of naturally occurring ω-aminoaldehydes including polyamine oxidation products. Recombinant isoenzymes from pea (PsAMADH1 and 2) and tomato (LeAMADH1 and 2) were subjected to kinetic measurements with synthetic aldehydes containing a nitrogenous heterocycle such as pyridinecarbaldehydes and their halogenated derivatives, (pyridinylmethylamino)-aldehydes, pyridinyl propanals and aldehydes derived from purine, 7-deazapurine and pyrimidine to characterize their substrate specificity and significance of the resulting data for in vivo reactions. The enzymatic production of the corresponding carboxylic acids was analyzed by liquid chromatography coupled to electrospray ionization mass spectrometry. Although the studied AMADHs are largely homologous and supposed to have a very similar active site architecture, significant differences were observed. LeAMADH1 displayed the broadest specificity oxidizing almost all compounds followed by PsAMADH2 and 1. In contrast, LeAMADH2 accepted only a few compounds as substrates. Pyridinyl propanals were converted by all isoenzymes, usually better than pyridinecarbaldehydes and aldehydes with fused rings. The K (m) values for the best substrates were in the range of 10(-5)-10(-4) M. Nevertheless, the catalytic efficiency values (V (max)/K (m)) reached only a very small fraction of that with 3-aminopropanal (except for LeAMADH1 activity with two pyridine-derived compounds). Docking experiments using the crystal structure of PsAMADH2 were involved to discuss differences in results with position isomers or alkyl chain homologs.

• MeSH
• aldehydoxidoreduktasy chemie   MeSH
• aldehydy chemie   MeSH
• aminokyselinové motivy MeSH
• heterocyklické sloučeniny chemie   MeSH
• hrách setý enzymologie   MeSH
• katalytická doména MeSH
• kinetika MeSH
• molekulární modely MeSH
• počítačová simulace MeSH
• rekombinantní proteiny chemie   MeSH
• rostlinné proteiny chemie   MeSH
• Solanum lycopersicum enzymologie   MeSH
• substrátová specifita MeSH
• vazba proteinů MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aldehydoxidoreduktasy MeSH
• aldehydy MeSH
• aminobutyraldehyde dehydrogenase MeSH Prohlížeč
• heterocyklické sloučeniny MeSH
• rekombinantní proteiny MeSH
• rostlinné proteiny MeSH

RATIONALE: Peroxidation of lipids in cellular membranes results in the release of volatile organic compounds (VOCs), including saturated aldehydes. The real-time quantification of trace VOCs produced by cancer cells during peroxidative stress presents a new challenge to non-invasive clinical diagnostics, which as described here, we have met with some success. METHODS: A combination of selected ion flow tube mass spectrometry (SIFT-MS), a technique that allows rapid, reliable quantification of VOCs in humid air and liquid headspace, and electrochemistry to generate reactive oxygen species (ROS) in vitro has been used. Thus, VOCs present in the headspace of CALU-1 cancer cell line cultures exposed to ROS have been monitored and quantified in real time using SIFT-MS. RESULTS: The CALU-1 lung cancer cells were cultured in 3D collagen to mimic in vivo tissue. Real-time SIFT-MS analyses focused on the volatile aldehydes: propanal, butanal, pentanal, hexanal, heptanal and malondialdehyde (propanedial), that are expected to be products of cellular membrane peroxidation. All six aldehydes were identified in the culture headspace, each reaching peak concentrations during the time of exposure to ROS and eventually reducing as the reactants were depleted in the culture. Pentanal and hexanal were the most abundant, reaching concentrations of a few hundred parts-per-billion by volume, ppbv, in the culture headspace. CONCLUSIONS: The results of these experiments demonstrate that peroxidation of cancer cells in vitro can be monitored and evaluated by direct real-time analysis of the volatile aldehydes produced. The combination of adopted methodology potentially has value for the study of other types of VOCs that may be produced by cellular damage.

• MeSH
• aldehydy analýza   metabolismus   MeSH
• buněčné kultury metody   MeSH
• elektrochemické techniky MeSH
• hmotnostní spektrometrie metody   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory metabolismus   MeSH
• oxidace-redukce MeSH
• oxidační stres fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldehydy MeSH

The Arabidopsis thaliana aldehyde oxidase 3 (AAO3) catalyzes the oxidation of abscisic aldehyde (ABal) to abscisic acid (ABA). Besides ABal, plants generate other aldehydes that can be toxic above a certain threshold. AAO3 knockout mutants (aao3) exhibited earlier senescence but equivalent relative water content compared with wild-type (WT) during normal growth or upon application of UV-C irradiation. Aldehyde profiling in leaves of 24-day-old plants revealed higher accumulation of acrolein, crotonaldehyde, 3Z-hexenal, hexanal and acetaldehyde in aao3 mutants compared with WT leaves. Similarly, higher levels of acrolein, benzaldehyde, crotonaldehyde, propionaldehyde, trans-2-hexenal and acetaldehyde were accumulated in aao3 mutants upon UV-C irradiation. Aldehydes application to plants hastened profuse senescence symptoms and higher accumulation of aldehydes, such as acrolein, benzaldehyde and 4-hydroxy-2-nonenal, in aao3 mutant leaves as compared with WT. The senescence symptoms included greater decrease in chlorophyll content and increase in transcript expression of the early senescence marker genes, Senescence-Related-Gene1, Stay-Green-Protein2 as well as NAC-LIKE, ACTIVATED-BY AP3/P1. Notably, although aao3 had lower ABA content than WT, members of the ABA-responding genes SnRKs were expressed at similar levels in aao3 and WT. Moreover, the other ABA-deficient mutants [aba2 and 9-cis-poxycarotenoid dioxygenase3-2 (nced3-2), that has functional AAO3] exhibited similar aldehydes accumulation and chlorophyll content like WT under normal growth conditions or UV-C irradiation. These results indicate that the absence of AAO3 oxidation activity and not the lower ABA and its associated function is responsible for the earlier senescence symptoms in aao3 mutant.

• Klíčová slova
• Arabidopsis, abscisic acid, aldehyde oxidase, reactive aldehydes, senescence,
• MeSH
• aldehydoxidasa genetika   metabolismus   MeSH
• aldehydy metabolismus   toxicita   MeSH
• Arabidopsis genetika   fyziologie   MeSH
• chlorofyl metabolismus   MeSH
• kyselina abscisová metabolismus   MeSH
• listy rostlin genetika   fyziologie   MeSH
• oxidace-redukce MeSH
• proteiny huseníčku genetika   metabolismus   MeSH
• regulátory růstu rostlin metabolismus   MeSH
• senescence rostlin MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• AAO3 protein, Arabidopsis MeSH Prohlížeč
• aldehydoxidasa MeSH
• aldehydy MeSH
• chlorofyl MeSH
• kyselina abscisová MeSH
• proteiny huseníčku MeSH
• regulátory růstu rostlin MeSH

Product ion distributions for the reactions of NO+ with 22 aldehydes involved in human physiology have been determined under the prevailing conditions of a selective reagent ionization time of flight mass spectrometry (SRI-TOF-MS) at an E/N in the flow/drift tube reactor of 130 Td. The chosen aldehydes were fourteen alkanals (the C2-C11 n-alkanals, 2-methyl propanal, 2-methyl butanal, 3-methyl butanal, and 2-ethyl hexanal), six alkenals (2-propenal, 2-methyl 2-propenal, 2-butenal, 3-methyl 2-butenal, 2-methyl 2-butenal, and 2-undecenal), benzaldehyde, and furfural. The product ion fragmentations patterns were determined for both dry air and humid air (3.5% absolute humidity) used as the matrix buffer/carrier gas in the drift tube of the SRI-TOF-MS instrument. Hydride ion transfer was seen to be a common ionization mechanism in all these aldehydes, thus generating (M-H)+ ions. Small fractions of the adduct ion, NO+M, were also seen for some of the unsaturated alkenals, in particular 2-undecenal, and heterocyclic furfural for which the major reactive channel was non-dissociative charge transfer generating the M+ parent ion. Almost all of the reactions resulted in partial fragmentation of the aldehyde molecules generating hydrocarbon ions; specifically, the alkanal reactions resulted in multiple product ions, whereas, the alkenals reactions produced only two or three product ions, dissociation of the nascent excited product ion occurring preferentially at the 2-position. The findings of this study are of particular importance for data interpretation in studies of aldehydes reactions employing SRI-TOF-MS in the NO+ mode.

• Klíčová slova
• Aldehydes, Fragmentation patterns, NO+ reactions, PTR-MS, SRI-TOF-MS, VOCs,
• Publikační typ
• časopisecké články MeSH

Due to their enhanced reactivity, metal and metal-oxide nanoscale zero-valent iron (nZVI) nanomaterials have been introduced into remediation practice. To ensure that environmental applications of nanomaterials are safe, their possible toxic effects should be described. However, there is still a lack of suitable toxicity tests that address the specific mode of action of nanoparticles, especially for nZVI. This contribution presents a novel approach for monitoring one of the most discussed adverse effects of nanoparticles, i.e., oxidative stress (OS). We optimized and developed an assay based on headspace-SPME-GC-MS analysis that enables the direct determination of volatile oxidative damage products (aldehydes) of lipids and proteins in microbial cultures after exposure to commercial types of nZVI. The method employs PDMS/DVB SPME fibers and pentafluorobenzyl derivatization, and the protocol was successfully tested using representatives of bacteria, fungi, and algae. Six aldehydes, namely, formaldehyde, acrolein, methional, benzaldehyde, glyoxal, and methylglyoxal, were detected in the cultures, and all of them exhibited dose-dependent sigmoidal responses. The presence of methional, which was detected in all cultures except those including an algal strain, documents that nZVI also caused oxidative damage to proteins in addition to lipids. The most sensitive toward nZVI exposure in terms of aldehyde production was the yeast strain Saccharomyces cerevisiae, which had an EC50 value of 0.08 g/L nZVI. To the best of our knowledge, this paper is the first to document the production of aldehydes resulting from lipids and proteins as a result of OS in microorganisms from different kingdoms after exposure to iron nanoparticles.

• Klíčová slova
• Nanomaterial, Oxidative stress, Remediation, SPME, Toxicity assay, Yeast,
• MeSH
• aldehydy farmakologie   MeSH
• Bacteria účinky léků   MeSH
• kovové nanočástice toxicita   MeSH
• lipidy MeSH
• nanostruktury MeSH
• oxidace-redukce MeSH
• oxidační stres fyziologie   MeSH
• testy toxicity MeSH
• železité sloučeniny MeSH
• železo metabolismus   toxicita   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldehydy MeSH
• ferric oxide MeSH Prohlížeč
• lipidy MeSH
• methional MeSH Prohlížeč
• železité sloučeniny MeSH
• železo MeSH

Nitric oxide plays an important role in the pathogenesis of Pseudoidium neolycopersici, the causative agent of tomato powdery mildew. S-nitrosoglutathione reductase, the key enzyme of S-nitrosothiol homeostasis, was investigated during plant development and following infection in three genotypes of Solanum spp. differing in their resistance to P. neolycopersici. Levels and localization of reactive nitrogen species (RNS) including NO, S-nitrosoglutathione (GSNO) and peroxynitrite were studied together with protein nitration and the activity of nitrate reductase (NR). GSNOR expression profiles and enzyme activities were modulated during plant development and important differences among Solanum spp. genotypes were observed, accompanied by modulation of NO, GSNO, peroxynitrite and nitrated proteins levels. GSNOR was down-regulated in infected plants, with exception of resistant S. habrochaites early after inoculation. Modulations of GSNOR activities in response to pathogen infection were found also on the systemic level in leaves above and below the inoculation site. Infection strongly increased NR activity and gene expression in resistant S. habrochaites in contrast to susceptible S. lycopersicum. Obtained data confirm the key role of GSNOR and modulations of RNS during plant development under normal conditions and point to their involvement in molecular mechanisms of tomato responses to biotrophic pathogens on local and systemic levels.

• Klíčová slova
• Nitric oxide, Powdery mildew, Pseudoidium neolycopersici, Reactive nitrogen species, S-nitrosoglutathione reductase, Solanum spp., Tomato,
• MeSH
• aldehydoxidoreduktasy metabolismus   MeSH
• Ascomycota patogenita   MeSH
• genotyp MeSH
• nemoci rostlin * mikrobiologie   MeSH
• reaktivní formy dusíku metabolismus   MeSH
• Solanum lycopersicum enzymologie   mikrobiologie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldehydoxidoreduktasy MeSH
• formaldehyde dehydrogenase, glutathione-independent MeSH Prohlížeč
• reaktivní formy dusíku MeSH

Aza-peptide aldehydes and ketones are a new class of reversible protease inhibitors that are specific for the proteasome and clan CD cysteine proteases. We designed and synthesised aza-Leu derivatives that were specific for the chymotrypsin-like active site of the proteasome, aza-Asp derivatives that were effective inhibitors of caspases-3 and -6, and aza-Asn derivatives that inhibited S. mansoni and I. ricinus legumains. The crystal structure of caspase-3 in complex with our caspase-specific aza-peptide methyl ketone inhibitor with an aza-Asp residue at P1 revealed a covalent linkage between the inhibitor carbonyl carbon and the active site cysteinyl sulphur. Aza-peptide aldehydes and ketones showed no cross-reactivity towards cathepsin B or chymotrypsin. The initial in vitro selectivity of these inhibitors makes them suitable candidates for further development into therapeutic agents to potentially treat multiple myeloma, neurodegenerative diseases, and parasitic infections.

• Klíčová slova
• Proteasome inhibitor, anticancer, antiparasitic, aza-peptide carbonyls, caspase and legumain inhibitors,
• MeSH
• aldehydy chemie   farmakologie   MeSH
• aza sloučeniny chemie   farmakologie   MeSH
• cysteinové endopeptidasy metabolismus   MeSH
• inhibitory proteas chemická syntéza   chemie   farmakologie   MeSH
• ketony chemie   farmakologie   MeSH
• krystalografie rentgenová MeSH
• lidé MeSH
• molekulární modely MeSH
• molekulární struktura MeSH
• peptidy chemie   farmakologie   MeSH
• proteasomový endopeptidasový komplex metabolismus   MeSH
• racionální návrh léčiv * MeSH
• skot MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aldehydy MeSH
• aza sloučeniny MeSH
• cysteinové endopeptidasy MeSH
• inhibitory proteas MeSH
• ketony MeSH
• peptidy MeSH
• proteasomový endopeptidasový komplex MeSH

Iron is necessary for life, but the simultaneous iron-catalyzed formation of reactive oxygen species (ROS) is involved in pathogenesis of many diseases. One of them is diabetes mellitus, a widespread disease with severe long-term complications, including neuropathy, retinopathy, and nephropathy. Much evidence points to methylglyoxal, a potent glycating agent, as the key mediator of diabetic complications. In diabetes, there is also a peculiar dysregulation of iron homeostasis, leading to an expansion of redox-active iron. This in vitro study focuses on the interaction of methylglyoxal with ferritin, which is the main cellular protein for iron storage. Methylglyoxal effectively liberates iron from horse spleen ferritin, as well as synthetic iron cores; in both cases, it is partially mediated by superoxide. The interaction of methylglyoxal with ferritin increases the production of hydrogen peroxide, much above the generation of peroxide by methylglyoxal alone, in an iron-dependent manner. Glycation with methylglyoxal results in structural changes in ferritin. All of these findings can be demonstrated with pathophysiologically relevant (submillimolar) methylglyoxal concentrations. However, the rate of iron release by ascorbate, the ferroxidase activity, or the diameter of gated pores even in intensely glycated ferritin is not altered. In conclusion, although the functional features of ferritin resist alterations due to glycation, the interaction of methylglyoxal with ferritin liberates iron and markedly increases ROS production, both of which could enhance oxidative stress in vivo. Our findings may have implications for the pathogenesis of long-term diabetic complications, as well as for the use of ferritin as a nanocarrier in chemotherapy.

Ferritin with methylglyoxal generates both Fenton reagents, ferrous iron, and hydrogen peroxide.Methylglyoxal produces structural modifications in ferritin that are detectable even in excess of another protein.Even heavily glycated ferritin has been found to resist functional alteration.

• Klíčová slova
• Glycation, diabetes mellitus, labile iron pool, protein cross-links, resistance to oxidative stress,
• MeSH
• ferritiny * metabolismus   MeSH
• koně MeSH
• lidé MeSH
• pyruvaldehyd * metabolismus   MeSH
• reaktivní formy kyslíku * metabolismus   MeSH
• železo metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ferritiny * MeSH
• pyruvaldehyd * MeSH
• reaktivní formy kyslíku * MeSH
• železo MeSH

Lipid peroxidation induced by oxidants leads to the formation of highly reactive metabolites. These can affect various immune functions, including reactive oxygen species (ROS) and nitric oxide (NO) production. The aim of the present study was to investigate the effects of lipid peroxidation products (LPPs) - acrolein, 4-hydroxynonenal, and malondialdehyde - on ROS and NO production in RAW 264.7 macrophages and to compare these effects with the cytotoxic properties of LPPs. Macrophages were stimulated with lipopolysaccharide (0.1 μg/ml) and treated with selected LPPs (concentration range: 0.1-100 μM). ATP test, luminol-enhanced chemiluminescence, Griess reaction, Western blotting analysis, amperometric and total peroxyl radical-trapping antioxidant parameter assay were used for determining the LPPs cytotoxicity, ROS and NO production, inducible nitric oxide synthase expression, NO scavenging, and antioxidant properties of LPPs, respectively. Our study shows that the cytotoxic action of acrolein and 4-hydroxynonenal works in a dose- and time-dependent manner. Further, our results imply that acrolein, 4-hydroxynonenal, and malondialdehyde can inhibit, to a different degree, ROS and NO production in stimulated macrophages, partially independently of their toxic effect. Also, changes in enzymatic pathways (especially NADPH-oxidase and nitric oxide synthase inhibition) and NO scavenging properties are included in the downregulation of reactive species formation.

• MeSH
• akrolein toxicita   MeSH
• aktivace makrofágů MeSH
• aldehydy toxicita   MeSH
• časové faktory MeSH
• down regulace účinky léků   MeSH
• dusitany metabolismus   MeSH
• lipopolysacharidy imunologie   MeSH
• makrofágy imunologie   metabolismus   MeSH
• malondialdehyd toxicita   MeSH
• myši MeSH
• NADPH-oxidasy antagonisté a inhibitory   MeSH
• osmolární koncentrace MeSH
• oxid dusnatý antagonisté a inhibitory   metabolismus   MeSH
• peroxidace lipidů * MeSH
• reaktivní formy kyslíku antagonisté a inhibitory   metabolismus   MeSH
• scavengery volných radikálů farmakologie   MeSH
• synthasa oxidu dusnatého, typ II metabolismus   MeSH
• transformované buněčné linie MeSH
• viabilita buněk účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• 4-hydroxy-2-nonenal MeSH Prohlížeč
• akrolein MeSH
• aldehydy MeSH
• dusitany MeSH
• lipopolysacharidy MeSH
• malondialdehyd MeSH
• NADPH-oxidasy MeSH
• Nos2 protein, mouse MeSH Prohlížeč
• oxid dusnatý MeSH
• reaktivní formy kyslíku MeSH
• scavengery volných radikálů MeSH
• synthasa oxidu dusnatého, typ II MeSH

(3,4-Dihydroxybut-1-ynyl)uracil, -cytosine and -7-deazaadenine 2'-deoxyribonucleoside triphosphates (dNTPs) were prepared by direct aqueous Sonogashira cross-coupling of halogenated dNTPs with dihydroxybut-1-yne and converted to 3,4-dihydroxybutyl dNTPs through catalytic hydrogenation. Sodium periodate oxidative cleavage of dihydroxybutyl-dUTP gave the desired aliphatic aldehyde-linked dUTP, whereas the oxidative cleavage of the corresponding deazaadenine dNTP gave a cyclic aminal. All dihydroxyalkyl or -alkynyl dNTPs and the formylethyl-dUTP were good substrates for DNA polymerases and were used for synthesis of diol- or aldehyde-linked DNA. The aldehyde linked DNA was used for the labelling or bioconjugations through hydrazone formation or reductive aminations.

• Klíčová slova
• DNA, aldehydes, bioconjugations, reactive groups, reductive amination,
• MeSH
• adenin analogy a deriváty   chemická syntéza   chemie   MeSH
• aldehydy chemie   MeSH
• aminace MeSH
• cytosin chemie   MeSH
• deoxyuracilnukleotidy chemická syntéza   chemie   MeSH
• DNA-dependentní DNA-polymerasy chemie   metabolismus   MeSH
• DNA chemie   metabolismus   MeSH
• molekulární struktura MeSH
• nukleotidy MeSH
• uracil chemie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 7-deazaadenine MeSH Prohlížeč
• adenin MeSH
• aldehydy MeSH
• cytosin MeSH
• deoxyuracilnukleotidy MeSH
• deoxyuridine triphosphate MeSH Prohlížeč
• DNA-dependentní DNA-polymerasy MeSH
• DNA MeSH
• nukleotidy MeSH
• uracil MeSH