Aroma profile and lactic acid bacteria characteristic of traditional Slovak cheese "May bryndza".

This paper describes the results of the characterization of a traditional Slovak cheese called "May bryndza" with regard to the profiles of volatile organic compounds and lactic acid bacteria. Samples of "May bryndza" cheese produced solely from unpasteurized ewe's milk were collected from 4 different Slovak farms, and samples of the cheese produced from a mixture of 2 types of milk (raw ewe's and pasteurized cow's milk) were collected from 3 different Slovak industrial dairies. There were 15 compounds detected and identified by the electronic nose. The impact of the kind of milk and the kind of dairy on the aroma profile of the product was not confirmed by PCA. The compounds with the highest relative contents in samples were acetoin (2.59%-24.55%), acetic acid (6.69%-13.39%), methoxy-phenyl-oxime (4.49%-8.52%), butanoic acid (1.89%-5.67%), and 2,3-butanediol (0.98%-4.08%), which were determined with gas chromatography. A total of 1533 isolates of LAB were obtained from the "May bryndza" cheese samples. Four families, five genera, and 19 species were identified with mass spectrometry, and isolated bacteria, both from the farm and industry dairies were the most frequently found to belong to Lactococcus lactis subsp. lactis.

Optimization of an Extraction Process to Obtain a Food-Grade Sulforaphane-Rich Extract from Broccoli (Brassica oleracea var. italica).

Sulforaphane (SFN) is a powerful health-promoting compound found in broccoli in the form of its inactive precursor, glucoraphanin (GFN). SFN formation occurs through the enzymatic hydrolysis of glucoraphanin by myrosinase under specific chemical conditions. Its incorporation in food formulations has been hindered by the thermal instability of SFN and low concentration in Brassicaceae. Then, extracting SFN from broccoli at a temperature below 40 °C appears as an option to recover and stabilize SFN, aiming at delivering it as a nutraceutical. We studied an eco-friendly extraction process to obtain an SFN-rich extract from broccoli. The effect of the broccoli mass/solvent ratio, ethanol concentration in the extractant solution, and extraction time on the recovery of SFN, GFN, phenolic compounds, and antioxidant activity were studied through a Box-Behnken design. The regression models explained more than 70% of the variability in the responses, adequately representing the system. The experimental factors differently affected the bioactive compound recovery and antioxidant activity of the extracts. The extraction conditions that allowed the highest recovery of bioactive compounds and antioxidant activity were identified and experimentally validated. The results may provide the basis for the design of a process to produce a sulforaphane-rich food supplement or nutraceutical by using a GRAS extractant.

The effect of processing and cooking on glucoraphanin and sulforaphane in brassica vegetables.

Brassica vegetables are widely consumed mostly after processing and cooking. These processing and cooking methods not only can affect the taste, texture, flavor and nutrients of these vegetables, but also influence the levels of some important bioactive compounds, such as glucosinolates (GLSs). Glucoraphanin (GLR) is the most abundant GLSs and its hydrolyzed component, sulforaphane (SLR), is the most powerful anti-cancer compound in brassica vegetables. In this review, we find out that varied treatments impact the retention of GLR and the formation of SLR differently. Be specific, 1) freezing can avoid the losses of GLR while short-time microwaving, short-time steaming and fermentation promote the biotransformation from GLR to SLR; 2) Boiling and blanching cause the largest losses of GLR and SLR, while freezing significantly protect their losses.; 3) Stir-frying varies the levels of GLR and SLR in different cooking conditions.

A novel high-performance liquid chromatography with diode array detector method for the simultaneous quantification of the enzyme-reactivating oximes obidoxime, pralidoxime, and HI-6 in human plasma.

Oximes such as pralidoxime (2-PAM), obidoxime (Obi), and HI-6 are the only currently available therapeutic agents to reactivate inhibited acetylcholinesterase (AChE) in case of intoxications with organophosphorus (OP) compounds. However, each oxime has characteristic agent-dependent reactivating efficacy, and therefore the combined administration of complementary oximes might be a promising approach to improve therapy. Accordingly, a new high-performance liquid chromatography method with diode-array detection (HPLC-DAD) was developed and validated allowing for simultaneous or single quantification of 2-PAM, Obi, and HI-6 in human plasma. Plasma was precipitated using 5% w/v aqueous zinc sulfate solution and subsequently acetonitrile yielding high recoveries of 94.2%-101.0%. An Atlantis T3 column (150 × 2.1mm I.D., 3 µm) was used for chromatographic separation with a total run time of 15 min. Quantification was possible without interferences within a linear range from 0.12 to 120 µg/mL for all oximes. Excellent intra-day (accuracy 91.7%-98.6%, precision 0.5%-4.4%) and inter-day characteristics (accuracy 89.4%-97.4%, precision 0.4%-2.2%) as well as good ruggedness were found. Oximes in processed samples were stable for at least 12 h in the autosampler at 15°C as well as in human plasma for at least four freeze-thaw cycles. Finally, the method was applied to plasma samples of a clinical case of pesticide poisoning.

Development of a relative quantification method for infrared matrix-assisted laser desorption electrospray ionization mass spectrometry imaging of Arabidopsis seedlings.

RATIONALE: Mass spectrometry imaging of young seedlings is an invaluable tool in understanding how mutations affect metabolite accumulation in plant development. However, due to numerous biological considerations, established methods for the relative quantification of analytes using infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry imaging are not viable options. In this study, we report a method for the quantification of auxin-related compounds using stable-isotope-labelled (SIL) indole-3-acetic acid (IAA) doped into agarose substrate. METHODS: Wild-type Arabidopsis thaliana seedlings, sur2 and wei8 tar2 loss-of-function mutants, and YUC1 gain-of-function line were grown for 3 days in the dark in standard growth medium. SIL-IAA was doped into a 1% low-melting-point agarose gel and seedlings were gently laid on top for IR-MALDESI imaging with Orbitrap mass spectrometry analysis. Relative quantification was performed post-acquisition by normalization of auxin-related compounds to SIL-IAA in the agarose. Amounts of auxin-related compounds were compared between genotypes to distinguish the effects of the mutations on the accumulation of indolic metabolites of interest. RESULTS: IAA added to agarose was found to remain stable, with repeatability and abundance features of IAA comparable with those of other compounds used in other methods for relative quantification in IR-MALDESI analyses. Indole-3-acetaldoxime was increased in sur2 mutants compared with wild-type and other mutants. Other auxin-related metabolites were either below the limits of quantification or successfully quantified but showing little difference among mutants. CONCLUSIONS: Agarose was shown to be an appropriate sampling surface for IR-MALDESI mass spectrometry imaging of Arabidopsis seedlings. SIL-IAA doping of agarose was demonstrated as a viable technique for relative quantification of metabolites in live seedlings or tissues with similar biological considerations.

Minor constituents of essential oils and aromatic extracts. Oximes derived from natural flavor and fragrance raw materials - Sensory evaluation, spectral and gas chromatographic characteristics.

A small library of 57 low molecular weight oximes was prepared from fragrant aldehydes and ketones, and their olfactory profiles were determined. The most substantive and interesting in terms of the sensory impressions were (+)-isomenthone oxime (fresh, musty, green) and cyclocitral oxime (earthy with patchouli, moss and leather notes). The linear retention indices (LRI) were determined for DB-1, DB-5 and DB-WAX columns, and E/Z isomers of 22 out of 57 compounds were resolved on the DB-5 column. Attempts were made to resolve enantiomers of the chosen oximes on chiral GC columns. The best results were obtained by using a Cyclosil B column, on which the enantiomers of camphor, menthone, piperitone and carvone oximes were fully resolved. NMR and MS spectra were acquired to characterize the synthesized library. Gas chromatography-olfactometry was used to assess odoriferous properties of both isomers of oximes. In most cases both isomers possessed similar profile and intensity.

Pharmacokinetics of K117 and K127, two novel antidote candidates to treat Tabun poisoning.

AIMS: K117 and K127 are bis-pyridinium aldoximes but K117 is a bis-pyridinium bis-aldoxime while K127 has only one single aldoxime in addition to its amide substituent. Is there any difference in pharmacokinetics in these compounds that otherwise have the same chemical structure? Both K117 and K127 are developed as antidotes in acetylcholinesterase and butyrylcholinesterase poisoning in terrorist attacks or intoxication with other organophosphorous compounds. Their distributions have been scouted in the bodies of rats. MAIN METHODS: White male Wistar rats were intramuscularly injected. The animals were sacrificed, tissue samples were homogenized, and either K117 or K127 concentrations were determined using reversed-phase high-performance liquid chromatography. KEY FINDINGS: Both K117 and K127 were present in all tissues that were analyzed including blood (serum), the brains, cerebrospinal fluid, the eyes, livers, kidneys, lungs and testes. Their pharmacokinetics and body distributions are similar. SIGNIFICANCE: Either K117 or K127 meets the essential requirements for antidotes. Dose dependence and kinetics of their distribution were compared to that of other pyridinium aldoximes.

99MTc-Hexamethylpropyleneamine Oxime Imaging for Early Detection of Acute Lung Injury in Rats Exposed to Hyperoxia or Lipopolysaccharide Treatment.

Tc-Hexamethylpropyleneamine oxime (HMPAO) is a clinical single-photon emission computed tomography biomarker of tissue oxidoreductive state. Our objective was to investigate whether HMPAO lung uptake can serve as a preclinical marker of lung injury in two well-established rat models of human acute lung injury (ALI).Rats were exposed to >95% O2 (hyperoxia) or treated with intratracheal lipopolysaccharide (LPS), with first endpoints obtained 24 h later. HMPAO was administered intravenously before and after treatment with the glutathione-depleting agent diethyl maleate (DEM), scintigraphy images were acquired, and HMPAO lung uptake was quantified from the images. We also measured breathing rates, heart rates, oxygen saturation, bronchoalveolar lavage (BAL) cell counts and protein, lung homogenate glutathione (GSH) content, and pulmonary vascular endothelial filtration coefficient (Kf).For hyperoxia rats, HMPAO lung uptake increased after 24 h (134%) and 48 h (172%) of exposure. For LPS-treated rats, HMPAO lung uptake increased (188%) 24 h after injury and fell with resolution of injury. DEM reduced HMPAO uptake in hyperoxia and LPS rats by a greater fraction than in normoxia rats. Both hyperoxia exposure (18%) and LPS treatment (26%) increased lung homogenate GSH content, which correlated strongly with HMPAO uptake. Neither of the treatments had an effect on Kf at 24 h. LPS-treated rats appeared healthy but exhibited mild tachypnea, BAL, and histological evidence of inflammation, and increased wet and dry lung weights. These results suggest the potential utility of HMPAO as a tool for detecting ALI at a phase likely to exhibit minimal clinical evidence of injury.

Micellar electrokinetic chromatographic method for the dabrafenib determination in biological samples.

Two different micellar electrokinetic chromatographic methods to determine dabrafenib in urine and serum, both using borate buffer (pH 9.2, 20 mM) and SDS as separation electrolyte, are developed and validated. The analyses were carried out in a fused-silica capillary of 75 µm of internal diameter and total length of 47 and 37 cm for urine and serum determination, respectively. The detection of the target compound was performed at 227 nm in urine samples and at 251 nm in serum samples. The linearity range was from 1 to 21 mg/L of dabrafenib in urine and from 2 to 40 mg/L in serum. In all cases, inter- and intraday RSDs were <4%. Sample preparation of serum samples consists of an only step of 1:1 dilution with water before its injection in the electrophoretic system. These simple, sensitive, accurate, and cost-effective methods can be used in routine clinical practice to monitor dabrafenib concentrations in urine and serum of metastatic melanoma skin cancer patients.

Development and validation of a FIA/UV-vis method for pK(a) determination of oxime based acetylcholinesterase reactivators.

Acetylcholinesterase reactivators (oximes) are compounds used for antidotal treatment in case of organophosphorus poisoning. The dissociation constants (pK(a1)) of ten standard or promising acetylcholinesterase reactivators were determined by ultraviolet absorption spectrometry. Two methods of spectra measurement (UV-vis spectrometry, FIA/UV-vis) were applied and compared. The soft and hard models for calculation of pK(a1) values were performed. The pK(a1) values were recommended in the range 7.00-8.35, where at least 10% of oximate anion is available for organophosphate reactivation. All tested oximes were found to have pK(a1) in this range. The FIA/UV-vis method provided rapid sample throughput, low sample consumption, high sensitivity and precision compared to standard UV-vis method. The hard calculation model was proposed as more accurate for pK(a1) calculation.

[Determination of Butroxydim in Agricultural Products by LC-MS].

An analytical method for the determination of butroxydim in agricultural products by LC-MS was developed. Butroxydim was extracted with acetonitrile and an aliquot of the crude extract was cleaned up on an octadecyl silanized silica gel (C18) cartridge column (1,000 mg), followed by a salting-out step to remove water. Before purification on a silica gel (SI) cartridge column (690 mg), polar matrices were precipitated by adding ethyl acetate, n-hexane and anhydrous sodium sulfate successively. This process effectively removed caffeine and catechins and improved recovery when analyzing residual butroxydim in tea leaves. Recovery and repeatability were good; the relative standard deviations were less than 5% for all 12 tested agricultural products (brown rice, soybean, potato, spinach, cabbage, apple, orange, grapefruit, lemon, tomato, peas with pods, and tea). Average recoveries for 11 agricultural products, except for lemon, were 74-92%.

[Indoor air guide values for 2-butanone oxime. Communication from the Ad-hoc Working Group on Indoor Guide Values of the Indoor Air Hygiene Commission and the States' Supreme Health Authorities]. / Richtwerte für Butanonoxim in der Innenraumluft : Mitteilung der Ad-hoc-Arbeitsgruppe Innenraumrichtwerte der Kommission Innenraumlufthygiene und der Obersten Landesgesundheitsbehörden.

The German Working Group on Indoor Guidelines of the Indoor Air Hygiene Committee and of the Supreme State Health Authorities is issuing indoor air guide values to protect public health. No reliable human studies are available for health evaluation of 2-butanone oxime in indoor air. In a well documented chronic inhalation animal study with rats and mice assessed as reliable, degenerative changes in the olfactory epithelium were observed, which led to a dose related increased incidence and severity, especially in mice. Using a benchmark approach the Working Group assessed a BMD10 of 13.8 mg 2-butanone oxime/m(3) for continuous exposure for the endpoint degeneration of the olfactory epithelium. For interspecies differences a reduced factor of 1 was applied due to the same susceptibility of rodents than human for this endpoint. By applying a factor of 10 for interindividual variability, and a factor of 2 to account for the higher respiratory rate of children compared to adults, a health hazard guide value (RW II) of 0.06 mg 2-butanone oxime/m(3) indoor air is obtained. A precautionary guide value of 0.02 mg 2-butanone oxime/m(3) indoor air is recommended.

Continuous water infusion enhances atmospheric pressure chemical ionization of methyl chloroformate derivatives in gas chromatography coupled to time-of-flight mass spectrometry-based metabolomics.

The effects of continuous water infusion on efficiency and repeatability of atmospheric pressure chemical ionization of both methyl chloroformate (MCF) and methoxime-trimethylsilyl (MO-TMS) derivatives of metabolites were evaluated using gas chromatography-time-of-flight mass spectrometry. Water infusion at a flow-rate of 0.4 mL/h yielded not only an average 16.6-fold increase in intensity of the quasimolecular ion for 20 MCF-derivatized metabolite standards through suppression of in-source fragmentation but also the most repeatable peak area integrals. The impact of water infusion was the greatest for dicarboxylic acids and the least for (hetero-) aromatic compounds. Water infusion also improved the ability to detect reliably fold changes as small as 1.33-fold for the same 20 MCF-derivatized metabolite standards spiked into a human serum extract. On the other hand, MO-TMS derivatives were not significantly affected by water infusion, neither in their fragmentation patterns nor with regard to the detection of differentially regulated compounds. As a proof of principle, we applied MCF derivatization and GC-APCI-TOFMS to the detection of changes in abundance of metabolites in pancreatic cancer cells upon treatment with 17-DMAG. Water infusion increased not only the number of metabolites identified via their quasimolecular ion but also the reproducibility of peak areas, thereby almost doubling the number of significantly regulated metabolites (false discovery rate < 0.05) to a total of 23.

The cost of nickel allergy: a global investigation of coin composition and nickel and cobalt release.

BACKGROUND: Nickel is widely used in coins; nickel may cause contact allergy and allergic contact dermatitis in those who handle them. OBJECTIVES: To investigate alloy use, coin composition and nickel and cobalt release for a worldwide selection of currently circulating coins. MATERIALS AND METHODS: Eight hundred and fifty coins of 361 different denominations or issues from 52 countries were collected and analysed with X-ray fluorescence spectrometry and nickel and cobalt spot tests. RESULTS: Copper-nickel was the most frequently identified coin alloy, being observed in 100 denominations (28%), followed by aluminium-bronze (62, 17%). In total, 239 denominations released nickel (28%). Coins from Bolivia, Brazil and Costa Rica did not release nickel. Fewer than one-third of the denominations or issues from China, India, the euro area and Indonesia released nickel. In the United States, the Russian Federation, Japan, and Mexico, one-third or more of the denominations released nickel. CONCLUSIONS: This worldwide selection of circulating coins covered countries with 75% of the world population, and shows that the majority of the world population lives in countries where coins release nickel. Pertinently, ∼ 40% of circulating coin denominations do not release nickel.

Test-retest stability of cerebral mGluR5 quantification using [¹¹C]ABP688 and positron emission tomography in rats.

This study evaluates the reproducibility of the quantification of metabotropic glutamate receptor type 5 (mGluR5) densities in rats using the PET radiotracer [¹¹C]ABP688 and pharmacokinetic models that are based on an input function, which is derived from a reference tissue. Seven rats underwent dynamic PET scans (60 min) after bolus injection of [¹¹C]ABP688. Kinetic analyses included: binding potential (BP(ND) ) determined by calculating (a) the simplified reference tissue model (SRTM) and (b) its two-steps simplified version (SRTM2); (c) multilinear reference tissue model (MRTM) and (d) its 2-parameter version (MRTM2); (e) noninvasive graphical analysis (NIGA). Parametric images were generated representing BP(ND) by the MRTM2 model. BP(ND) determinations were reproducible with low to acceptable variability ranging from 5 to 10% and reproducibility scores (intraclass correlation coefficient) between 0.51 and 0.88. The pharmacokinetic model that showed lowest overall variability was the SRTM. In contrast, the use of the NIGA was associated with significantly lower reproducibility scores. Comparison of parametric images revealed no significant bias between test and retest measurements and is therefore suitable to compare groups at voxel levels. In conclusion, our results suggest that noninvasive quantification of [¹¹C]ABP688 imaging is reproducible and reliable for PET studies of the cerebral mGluR5 in rats.

Oxime-based carbonates as useful reagents for both N-protection and peptide coupling.

We have demonstrated that oxime-based mixed carbonates are very effective reagents for both N-protection and peptide coupling.

Synthesis and screening of aromatase inhibitory activity of substituted C19 steroidal 17-oxime analogs.

The synthesis and aromatase inhibitory activity of androst-4-en-, androst-5-en-, 1ß,2ß-epoxy- and/or androsta-4,6-dien-, 4ß,5ß-epoxyandrostane-, and 4-substituted androst-4-en-17-oxime derivatives are described. Inhibition activity of synthesized compounds was assessed using aromatase enzyme and [1ß-3H]androstenedione as substrate. Most of the compounds displayed similar to or more aromatase inhibitory activity than formestane (74.2%). 4-Chloro-3ß-hydroxy-4-androsten-17-one oxime (14, 93.8%) showed the highest activity, while 4-azido-3ß-hydroxy-4-androsten-17-one oxime (17, 32.8%) showed the lowest inhibitory activity for aromatase.

Z-formamidoximes in molecular folding and macrocycles.

The formamidoxime configurational Z isomer coupled with the pyridylbiscarboxamide conformational codon were used to fold planar, curved structures. When embedded into macrocycles, this folded motif promotes dimerization through π-π stacking and hydrogen-bonding and the formation of tubules akin to molecular channels in the solid state.

Antigen heterologous enzyme linked immunosorbent assay for the measurement of DHEA in serum.

Homologous and heterologous combinations of enzyme conjugate and antibody in steroid enzyme immunoassay (EIA) influences unlabeled steroid recognition by antibody that affects sensitivity of the assay. To develop dehydroepiandrosterone (DHEA) antigen heterologous enzyme linked immunosorbent assay (ELISA), antibodies were generated against DHEA-3-hemisuccinate-bovine serum albumin (DHEA-3-HS-BSA), DHEA-7-carboxymethyloxime-bovine serum albumin (DHEA-7-CMO-BSA), and DHEA-17-carboxymethyloxime-bovine serum albumin (DHEA-17-CMO-BSA). Five horseradish peroxidase (HRP) enzyme conjugates were prepared using five testosterone derivatives [testosterone-3-CMO (T-3-CMO), testosterone-17-HS (T-17-HS), testosterone-17-glucuronoside (T-17-G), testosterone-19-carboxymethylether (T-19-CME), and testosterone-11-HS (T-11-HS)]. Fifteen antigen heterologous combinations of antibody and enzyme conjugates were evaluated in the standard binding assay; only two combinations showed binding. The use of antigen heterologous combination (different antigen in label than the immunogen) resulted in development of a simple, direct, and convenient assay as it permits the direct addition of the serum sample into the assay and it requires only 1.5 h to complete.