• Klíčová slova
• INDUSTRIAL MEDICINE *, WAXES *,
• MeSH
• lidé MeSH
• pracovní lékařství * MeSH
• vosky * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• vosky * MeSH

Lithium salts of organic aromatic acids (lithium benzoate, lithium salicylate, lithium vanillate, lithium 2,5-dimethoxybenzoate, lithium 2,5-dihydroxyterephthalate, lithium α-cyano-4-hydroxycinnamate and lithium sinapate) were synthesized and tested as potential matrices for the matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry analysis of hydrocarbons and wax esters. The analytes were desorbed using nitrogen laser (337.1 nm) and ionized via the attachment of a lithium cation, yielding [M + Li](+) adducts. The sample preparation and the experimental conditions were optimized for each matrix using stearyl behenate and n-triacontane standards. The performance of the new matrices in terms of signal intensity and reproducibility, the mass range occupied by matrix ions and the laser power threshold were studied and compared with a previously recommended lithium 2,5-dihydroxybenzoate matrix (LiDHB) (Cvačka and Svatoš, Rapid Commun. Mass Spectrom. 2003, 17, 2203). Several of the new matrices performed better than LiDHB. Lithium vanillate offered a 2-3 times and 7-9 times higher signal for wax esters and hydrocarbons, respectively. Also, the signal reproducibility improved substantially, making this matrix a suitable candidate for imaging applications. In addition, the diffuse reflectance spectra and solubility of the synthesized compounds were investigated and discussed with respect to the compound's ability to serve as MALDI matrices. The applicability of selected matrices was tested on natural samples of wax esters and hydrocarbons.

• Klíčová slova
• MALDI matrix, cuticular hydrocarbons, lipids, lithium attachment, waxes,
• MeSH
• lithium chemie   MeSH
• Sarcophagidae chemie   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• uhlovodíky analýza   chemie   MeSH
• vosky analýza   chemie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• lithium MeSH
• uhlovodíky MeSH
• vosky MeSH

Wax esters (WEs), esters of long-chain fatty acids and long-chain alcohols, were analysed by Ag-HPLC/APCI-MS/MS. Two ChromSpher Lipids columns connected in series (a total length of 50cm) and hexane-2-propanol-acetonitrile mobile phases were used to achieve good separation of the molecular species. The chromatographic behaviour of WEs was studied under optimised conditions: retention increased with the number of double bonds and with the temperature (15-35°C); retention times were affected by the double-bond position, trans isomers eluted earlier than cis isomers, and the WEs were partially separated depending on the aliphatic-chain length. The WEs provided simple APCI spectra with [M+H](+) ions, the MS/MS spectra showed fragments, which allowed their identification. The method was applied for an analysis of the WE mixtures from jojoba oil and human hair and the results were compared with analogous data from an optimised RP-HPLC system.

• Klíčová slova
• Human hair, Jojoba, Long-chain esters, Mass spectrometry, Silver-ion liquid chromatography, Wax esters,
• MeSH
• estery analýza   MeSH
• lidé MeSH
• stříbro chemie   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• vosky analýza   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• estery MeSH
• jojoba wax MeSH Prohlížeč
• stříbro MeSH
• vosky MeSH

We suggest a new technique for estimating the relative drawdown of CO2 concentration (c) in the intercellular air space (IAS) across hypostomatous leaves (expressed as the ratio cd/cb, where the indexes d and b denote the adaxial and abaxial edges, respectively, of IAS), based on the carbon isotope composition (δ13C) of leaf cuticular membranes (CMs), cuticular waxes (WXs) or epicuticular waxes (EWXs) isolated from opposite leaf sides. The relative drawdown in the intracellular liquid phase (i.e., the ratio cc/cbd, where cc and cbd stand for mean CO2 concentrations in chloroplasts and in the IAS), the fraction of intercellular resistance in the total mesophyll resistance (rIAS/rm), leaf thickness, and leaf mass per area (LMA) were also assessed. We show in a conceptual model that the upper (adaxial) side of a hypostomatous leaf should be enriched in 13C compared to the lower (abaxial) side. CM, WX, and/or EWX isolated from 40 hypostomatous C3 species were 13C depleted relative to bulk leaf tissue by 2.01-2.85‰. The difference in δ13C between the abaxial and adaxial leaf sides (δ13CAB - 13CAD, Δb-d), ranged from - 2.22 to + 0.71‰ (- 0.09 ± 0.54‰, mean ± SD) in CM and from - 7.95 to 0.89‰ (- 1.17 ± 1.40‰) in WX. In contrast, two tested amphistomatous species showed no significant Δb-d difference in WX. Δb-d correlated negatively with LMA and leaf thickness of hypostomatous leaves, which indicates that the mesophyll air space imposes a non-negligible resistance to CO2 diffusion. δ13C of EWX and 30-C aldehyde in WX reveal a stronger CO2 drawdown than bulk WX or CM. Mean values of cd/cb and cc/cbd were 0.90 ± 0.12 and 0.66 ± 0.11, respectively, across 14 investigated species in which wax was isolated and analyzed. The diffusion resistance of IAS contributed 20 ± 14% to total mesophyll resistance and reflects species-specific and environmentally-induced differences in leaf functional anatomy.

• Klíčová slova
• CO2 diffusion, Carbon isotope discrimination, Cuticle, Leaf mesophyll, Leaf traits, Mesophyll conductance, Waxes,
• MeSH
• biologické modely MeSH
• extracelulární prostor metabolismus   MeSH
• izotopy uhlíku metabolismus   MeSH
• listy rostlin anatomie a histologie   růst a vývoj   metabolismus   MeSH
• mezofylové buňky metabolismus   MeSH
• nadmořská výška MeSH
• oxid uhličitý metabolismus   MeSH
• vosky metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• izotopy uhlíku MeSH
• oxid uhličitý MeSH
• vosky MeSH

Higher wax esters within the range of C24 to C44 (205 standards) were analyzed by means of gas chromatography and Kováts indexes (I) and reduced Kováts indexes (RKI) were calculated. The dependences of these retention data on number of carbon atoms and on number and position of double bonds in acid and in alcohol moieties of esters were plotted.

• MeSH
• chromatografie plynová metody   MeSH
• estery analýza   chemie   MeSH
• mastné kyseliny chemie   MeSH
• vosky analýza   chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• estery MeSH
• mastné kyseliny MeSH
• vosky MeSH

The interpretation of the electron ionization mass spectra of straight-chain and methyl-branched saturated and unsaturated wax esters (WEs) is discussed in this study based on the spectra of 154 standards. The most important fragments indicative of the structure of the acid and alcohol chains are identified and summarized for WEs with various number of double bonds in the chains. Briefly, most WEs provide acylium ions allowing structural characterization of the acid part, whereas the alcohol part gives corresponding alkyl radical cations. The elemental composition of selected important fragments is established from a high-resolution accurate mass analysis. The ion abundances are discussed with respect to the length and unsaturation of the aliphatic chains. The interpretation of the spectra of branched or unsaturated WEs requires the recognition of small but important peaks that are difficult to discern among the other fragments. We demonstrate that such fragments are easily detected in differential mass spectra. This approach requires spectra of WE standards (e.g., straight-chain analogs in the case of branched WEs) recorded under the same experimental conditions. The WEs mass spectral database provided in the supplemental data can be used as a reference for the analysis of the GC/EI-MS data.

• MeSH
• estery chemie   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• nenasycené mastné kyseliny chemie   MeSH
• plynová chromatografie s hmotnostně spektrometrickou detekcí metody   MeSH
• referenční standardy MeSH
• vosky chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• estery MeSH
• nenasycené mastné kyseliny MeSH
• vosky MeSH

BACKGROUND AND AIMS: The benefits and costs of amphistomy (AS) vs. hypostomy (HS) are not fully understood. Here, we quantify benefits of access of CO2 through stomata on the upper (adaxial) leaf surface, using 13C abundance in the adaxial and abaxial epicuticular wax. Additionally, a relationship between the distribution of stomata and epicuticular wax on the opposite leaf sides is studied. METHODS: We suggest that the 13C content of long-chain aliphatic compounds of cuticular wax records the leaf internal CO2 concentration in chloroplasts adjacent to the adaxial and abaxial epidermes. This unique property stems from: (1) wax synthesis being located exclusively in epidermal cells; and (2) ongoing wax renewal over the whole leaf lifespan. Compound-specific and bulk wax 13C abundance (δ) was related to amphistomy level (ASL; as a fraction of adaxial in all stomata) of four AS and five HS species grown under various levels of irradiance. The isotopic polarity of epicuticular wax, i.e. the difference in abaxial and adaxial δ (δab - δad), was used to calculate the leaf dorsiventral CO2 gradient. Leaf-side-specific epicuticular wax deposition (amphiwaxy level) was estimated and related to ASL. KEY RESULTS: In HS species, the CO2 concentration in the adaxial epidermis was lower than in the abaxial one, independently of light conditions. In AS leaves grown in high-light and low-light conditions, the isotopic polarity and CO2 gradient varied in parallel with ASL. The AS leaves grown in high-light conditions increased ASL compared with low light, and δab - δad approached near-zero values. Changes in ASL occurred concomitantly with changes in amphiwaxy level. CONCLUSIONS: Leaf wax isotopic polarity is a newly identified leaf trait, distinguishing between hypo- and amphistomatous species and indicating that increased ASL in sun-exposed AS leaves reduces the CO2 gradient across the leaf mesophyll. Stomata and epicuticular wax deposition follow similar leaf-side patterning.

• Klíčová slova
• Brassica oleracea, Capsicum annuum, Amphistomy, abaxial, adaxial, carbon isotope, cuticle, epicuticular wax, leaf internal CO2 concentration, light, photosynthesis, stomata,
• MeSH
• epidermis rostlin * metabolismus   MeSH
• fotosyntéza MeSH
• izotopy uhlíku * analýza   MeSH
• listy rostlin * metabolismus   MeSH
• oxid uhličitý * metabolismus   MeSH
• průduchy rostlin * fyziologie   MeSH
• vosky * metabolismus   chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• Carbon-13 MeSH Prohlížeč
• izotopy uhlíku * MeSH
• oxid uhličitý * MeSH
• vosky * MeSH

Signals of different nature are transduced in cells through signal transduction pathways, where mitogen-activated protein kinases (MAPKs) play an important role as signaling molecules. Views into intracellular localization of MAPKs are critical for the understanding of their spatial and temporal functions, like activation-based relocation, compartmentation, or interactions with local substrates. Localization of MAPKs in cells is thus very useful cell biological approach, extending complex mode of cell signaling characterization in plants. Here, we present a method for subcellular immunofluorescence localization of MAPKs using protein- or phospho-specific antibodies, performed on sectioned fixed plant samples. It is based on embedding of samples in the Steedman's wax, a low-melting point polyester wax embedding medium, which maintains high antigenicity of studied proteins. In addition, exposure of dewaxed sections to antibodies allows for their efficient penetration. Altogether, it makes this simple method a good tool in the efficient subcellular localization of diverse proteins, including plant MAPKs.

• MeSH
• Arabidopsis cytologie   účinky léků   enzymologie   MeSH
• fluorescenční mikroskopie MeSH
• fluorescenční protilátková technika metody   MeSH
• Medicago sativa cytologie   účinky léků   enzymologie   MeSH
• mitogenem aktivované proteinkinasy analýza   metabolismus   MeSH
• polyestery * MeSH
• transport proteinů MeSH
• vosky * MeSH
• zalévání tkání metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• mitogenem aktivované proteinkinasy MeSH
• polyestery * MeSH
• vosky * MeSH

The plant cuticle is an important plant-atmosphere boundary, the synthesis and maintenance of which represents a significant metabolic cost. Only limited information regarding cuticle dynamics is available. We determined the composition and dynamics of Clusia rosea cuticular waxes and matrix using 13 CO2 labelling, compound-specific and bulk isotope ratio mass spectrometry. Collodion was used for wax collection; gas exchange techniques to test for any collodion effects on living leaves. Cutin matrix (MX) area density did not vary between young and mature leaves and between leaf sides. Only young leaves incorporated new carbon into their MX. Collodion-based sampling discriminated between epicuticular (EW) and intracuticular wax (IW) effectively. Epicuticular differed in composition from IW. The newly synthetised wax was deposited in IW first and later in EW. Both young and mature leaves synthetised IW and EW. The faster dynamics in young leaves were due to lower wax coverage, not a faster synthesis rate. Longer-chain alkanes were deposited preferentially on the abaxial, stomatous leaf side, producing differences between leaf sides in wax composition. We introduce a new, sensitive isotope labelling method and demonstrate that cuticular wax is renewed during leaf ontogeny of C. rosea. We discuss the ecophysiological significance of the new insights.

• Klíčová slova
• 13C labelling, cutin, development, gas exchange, photosynthesis, plant cuticle, wax regeneration,
• MeSH
• Clusia * metabolismus   MeSH
• epidermis rostlin metabolismus   MeSH
• kolodium analýza   metabolismus   MeSH
• listy rostlin fyziologie   MeSH
• oxid uhličitý * metabolismus   MeSH
• vosky metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kolodium MeSH
• oxid uhličitý * MeSH
• vosky MeSH

The objective of this work was to create a fast and sensitive method of detecting Paenibacillus larvae from beehive debris based on PCR that does not require long-lasting cultivation steps. Various methods of extracting spores from beehive debris were compared: the original method of extraction of spores into toluene, and alternative spore extraction methods into Tween 80, into water, into isopropanol and into 95% ethanol. Isolation of DNA from various spore extractions was evaluated too. Best results were provided by isolation of DNA using the QIAamp DNA Mini Kit, without heat treatment. DNA of spores was detected by PCR from 0.25 g of beeswax debris, with the detected titer of 10(5) in 1g according to the cultivation tests.

• MeSH
• DNA bakterií genetika   MeSH
• kolaps kolonií mikrobiologie   MeSH
• Paenibacillus genetika   MeSH
• polymerázová řetězová reakce metody   MeSH
• spory bakteriální genetika   MeSH
• včely mikrobiologie   MeSH
• vosky MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA bakterií MeSH
• vosky MeSH