Interaction between diterpene icetexanes and old yellow enzymes of Leishmania braziliensis and Trypanosoma cruzi.

Old Yellow Enzymes (OYEs) are flavin-dependent redox enzymes that promote the asymmetric reduction of activated alkenes. Due to the high importance of flavoenzymes in the metabolism of organisms, the interaction between OYEs from the parasites Trypanosoma cruzi and Leishmania braziliensis and three diterpene icetexanes (brussonol and two analogs), were evaluated in the present study, and differences in the binding mechanism and inhibition capacity of these molecules were examined. Although the aforementioned compounds showed poor and negligible activities against T. cruzi and L. braziliensis cells, respectively, the experiments with the purified enzymes indicated that the interaction occurs by divergent mechanisms. Overall, the ligands' inhibitory effect depends on their accessibility to the N5 position of the flavin's isoalloxazine ring. The results also indicated that the OYEs found in both parasites share structural similarities and showed affinities for the diterpene icetexanes in the same range. Nevertheless, the interaction between OYEs and ligands is directed by enthalpy and/or entropy in distinct ways. In conclusion, the binding site of both OYEs exhibits remarkable plasticity, and a large range of different molecules, including that can be substrates and inhibitors, can bind this site. This plasticity should be considered in drug design using OYE as a target.

Impact of Physicochemical Modifications in Casein Promoted by UV-C on the Peptide Profile of Gastric Digestion and the Transepithelial Transport of Peptides.

Caseins are the main proteins in milk, and their structure and spatial conformation are responsible for their slow digestion rate. The release of bioactive and ß-casomorphin peptides from casein digestion may induce allergic responses during consumption. Spectroscopic techniques were used to observe the structural changes in casein conformation induced by Ultraviolet light irradiation (UV-C). Raman spectroscopy results showed more pronounced peaks at 618 and 640 cm-1 for phenylalanine and tyrosine moieties of the photolyzed micellar casein, respectively, suggesting changes in the micelle structure. The decrease in the intensity of Raman signals for tryptophan and tyrosine corroborates to the UV-C-induced modifications of the micelle structure. Particle size distribution showed a decrease in the average micelle size after 15 min of UV-C exposure, while low-temperature, long-time (LTLT) pasteurization led to the formation of large aggregates, as observed by atomic force microscopy. UV-C did not impact the formation or transport of peptides, as observed by using the Caco-2 cell as a model for peptide absorption. However, the absence of the opioid peptide SRYPSY from κ-casein and only 20% of the concentration of opioid peptide RYLGY were noted. This work demonstrated that UV-C can be utilized to induce the physicochemical modification of dairy products, promoting a higher digestion rate and reducing allergenicity.

Organ-on-a-Chip for Drug Screening: A Bright Future for Sustainability? A Critical Review.

Globalization has raised concerns about spreading diseases and emphasized the need for quick and efficient methods for drug screening. Established drug efficacy and toxicity approaches have proven obsolete, with a high failure rate in clinical trials. Organ-on-a-chip has emerged as an essential alternative to outdated techniques, precisely simulating important characteristics of organs and predicting drug pharmacokinetics more ethically and efficiently. Although promising, most organ-on-a-chip devices are still manufactured using principles and materials from the micromachining industry. The abusive use of plastic for traditional drug screening methods and device production should be considered when substituting technologies so that the compensation for the generation of plastic waste can be projected. This critical review outlines recent advances for organ-on-a-chip in the industry and estimates the possibility of scaling up its production. Moreover, it analyzes trends in organ-on-a-chip publications and provides suggestions for a more sustainable future for organ-on-a-chip research and production.

UV-C light promotes the reductive cleavage of disulfide bonds in ß-Lactoglobulin and improves in vitro gastric digestion.

ß-Lactoglobulin (ß-Lg) is the main protein in whey and is known for its allergenicity and resistance to the digestion of pepsin and trypsin. The UV-C photoinduced cleavage of disulfide bonds in ß-Lactoglobulin, as promoted by excitation of tryptophan residues (Trp), is shown to induce changes in the protein's secondary structure, significantly reducing the protein's resistance to pepsin digestion. The UV-C light-induced changes in the protein secondary structure are marked by an increase in the contribution of ß-sheet and α-helix structures with a concomitantly smaller contribution of the ß-turn structural motif. The photoinduced cleavage of disulfide bonds in ß-Lg has an apparent quantum yield of ф = 0.0015 ± 0.0003 and was shown by transient absorption laser flash photolysis to arise by two different pathways: a) the reduction of the disulfide bond Cys66Cys160 occurs by direct electron transfer from the triplet-excited 3Trp to the disulfide bond due to the existence of a CysCys/Trp triad (Cys66Cys160/Trp61) and b) the reduction of the buried Cys106Cys119 disulfide bond involves a reaction with a solvated electron originated by the photoejection of electrons from the triplet-excited 3Trp decay. The in vitro gastric digestion index for UV-C-treated ß-Lg is revealed to have increased significantly by 36 ± 4 % and 9 ± 2 % under simulated elderly and young adult digestive conditions, respectively. When compared to the native protein, the peptide mass fingerprint profile of digested UV-C-treated ß-Lg shows a higher content and variety of peptides, including the production of some exclusive bioactive peptides such as PMHIRL and EKFDKALKALPMH.

Unraveling Hepatic Metabolomic Profiles and Morphological Outcomes in a Hybrid Model of NASH in Different Mouse Strains.

Nonalcoholic fatty liver disease (NAFLD) encompasses nonalcoholic steatohepatitis (NASH) and affects 25% of the global population. Although a plethora of experimental models for studying NASH have been proposed, still scarce findings regarding the hepatic metabolomic/molecular profile. In the present study, we sought to unravel the hepatic metabolomic profile of mice subjected to a hybrid model of NASH, by combining a Western diet and carbon tetrachloride administration, for 8 weeks, in male C57BL/6J and BALB/c mice. In both mouse strains, the main traits of NASH-metabolic (glucose intolerance profile), morphologic (extensive microvesicular steatosis and fibrosis, lobular inflammation, and adipose tissue-related inflammation/hypertrophy), and molecular (impaired Nrf2/NF-κB pathway dynamics and altered metabolomic profile)-were observed. The hepatic metabolomic profile revealed that the hybrid protocol impaired, in both strains, the abundance of branched chain-aromatic amino acids, carboxylic acids, and glycosyl compounds, that might be linked to the Nrf2 pathway activation. Moreover, we observed a strain-dependent hepatic metabolomic signature, in which the tricarboxylic acid metabolites and pyruvate metabolism were dissimilarly modulated in C57BL/6J and BALB/c mice. Thus, we provide evidence that the strain-dependent hepatic metabolomic profile might be linked to the distinct underlying mechanisms of NASH, also prospecting potential mechanistic insights into the corresponding disease.

Are isothiocyanates and polyphenols from Brassicaceae vegetables emerging as preventive/therapeutic strategies for NAFLD? The landscape of recent preclinical findings.

Nonalcoholic fatty liver disease (NAFLD) is a lipid impairment-related chronic metabolic disease that affects almost 25% of the worldwide population and has become the leading cause of liver transplantation in the United States of America (USA). NAFLD may progress from simple hepatic steatosis (HS) to nonalcoholic steatohepatitis (NASH), which occurs simultaneously in an inflammatory and fibrotic microenvironment and affects approximately 5% of the global population. Recently, NASH has been suggested to be a relevant driver in progressive liver cirrhosis and a population-attributable factor in hepatocellular carcinoma patients. Moreover, predictions show that NAFLD-related annual health costs in the USA have reached ∼$100 bi., but effective therapies are still scarce. Thus, new preventative strategies for this hepatic disease urgently need to be developed. The Brassicaceae vegetable family includes almost 350 genera and 3500 species and these are one of the main types of vegetables harvested and produced worldwide. These vegetables are well-known sources of glucobrassicin-derivative molecules, such as isothiocyanates and phenolic compounds, which have shown antioxidant and antilipogenic effects in preclinical NAFLD data. In this review, we gathered prominent evidence of the in vivo and in vitro effects of these vegetable-derived nutraceutical compounds on the gut-liver-adipose axis, which is a well-known regulator of NAFLD and may represent a new strategy for disease control.

1H qNMR-Based Metabolomics Discrimination of Covid-19 Severity.

The coronavirus disease 2019 (Covid-19), which caused respiratory problems in many patients worldwide, led to more than 5 million deaths by the end of 2021. Experienced symptoms vary from mild to severe illness. Understanding the infection severity to reach a better prognosis could be useful to the clinics, and one study area to fulfill one piece of this biological puzzle is metabolomics. The metabolite profile and/or levels being monitored can help predict phenotype properties. Therefore, this study evaluated plasma metabolomes of 110 individual samples, 57 from control patients and 53 from recent positive cases of Covid-19 (IgM 98% reagent), representing mild to severe symptoms, before any clinical intervention. Polar metabolites from plasma samples were analyzed by quantitative 1H NMR. Glycerol, 3-aminoisobutyrate, formate, and glucuronate levels showed alterations in Covid-19 patients compared to those in the control group (Tukey's HSD p-value cutoff = 0.05), affecting the lactate, phenylalanine, tyrosine, and tryptophan biosynthesis and d-glutamine, d-glutamate, and glycerolipid metabolisms. These metabolic alterations show that SARS-CoV-2 infection led to disturbance in the energetic system, supporting the viral replication and corroborating with the severe clinical conditions of patients. Six polar metabolites (glycerol, acetate, 3-aminoisobutyrate, formate, glucuronate, and lactate) were revealed by PLS-DA and predicted by ROC curves and ANOVA to be potential prognostic metabolite panels for Covid-19 and considered clinically relevant for predicting infection severity due to their straight roles in the lipid and energy metabolism. Thus, metabolomics from samples of Covid-19 patients is a powerful tool for a better understanding of the disease mechanism of action and metabolic consequences of the infection in the human body and may corroborate allowing clinicians to intervene quickly according to the needs of Covid-19 patients.

Effects of dietary inclusion of yerba mate (Ilex paraguariensis) extract on lamb muscle metabolomics and physicochemical properties in meat.

This study aimed to evaluate the effect of dietary yerba mate (Ilex paraguariensis) extract (YME) on muscle metabolomics and physicochemical properties of lamb meat. Thirty-six uncastrated male lambs (90 d old) were fed experimental diets, which treatments consisted of 0%, 1%, 2%, and 4% inclusion of YME. Animals were fed for 50 d before slaughter. Muscle and meat samples were collected for metabolomics and meat quality analysis, respectively. The experiment was carried out in a randomized block design and analyzed using orthogonal contrasts. There was a quadratic effect of YME inclusion in tenderness (P < 0.05) and a positive linear effect on meat lightness (P < 0.05). No qualitative changes (P > 0.05) on individual metabolites were observed; however, changes in the quantitative metabolic profile were observed, showing that animals fed 1% and 2% of YME have a greater concentration of desirable endogenous muscle antioxidants, with direct impact on metabolic pathways related to beta-alanine metabolism and glutathione metabolism. Therefore, YME dietary supplementation up to 2% of the diet to lambs had little to no effects on the majority of meat quality traits evaluated; moreover, 4% of YME inclusion negatively affected feed intake and meat quality traits.

Light-activated generation of nitric oxide (NO) and sulfite anion radicals (SO3˙-) from a ruthenium(ii) nitrosylsulphito complex.

This manuscript describes the preparation of a new Ru(ii) nitrosylsulphito complex, trans-[Ru(NH3)4(isn)(N(O)SO3)]+ (complex 1), its spectroscopic and structural characterization, photochemistry, and thermal reactivity. Complex 1 was obtained by the reaction of sulfite ions (SO32-) with the nitrosyl complex trans-[Ru(NH3)4(isn)(NO)]3+ (complex 2) in aqueous solution resulting in the formation of the N-bonded nitrosylsulphito (N(O)SO3) ligand. To the best of our knowledge, only four nitrosylsulphito metal complexes have been described so far (J. Chem. Soc., Dalton Trans., 1983, 2465-2472), and there is no information about the photochemistry of such complexes. Complex 1 was characterized by spectroscopic means (UV-Vis, EPR, FT-IR, 1H- and 15N-NMR), elemental analysis and single-crystal X-ray diffraction. The X-ray structure of the precursor complex 2 is also discussed in the manuscript and is used as a reference for comparisons with the structure of 1. Complex 1 is water-soluble and kinetically stable at pH 7.4, with a first-order rate constant of 3.1 × 10-5 s-1 for isn labilization at 298 K (t1/2∼ 373 min). Under acidic conditions (1.0 M trifluoroacetic acid), 1 is stoichiometrically converted into the precursor complex 2. The reaction of hydroxide ions (OH-) with 1 and with 2 yields the Ru(ii) nitro complex trans-[Ru(NH3)4(isn)(NO2)]+ with second-order rate constants of 2.1 and 10.5 M-1 s-1 (at 288 K), respectively, showing the nucleophilic attack of OH- at the nitrosyl in 2 (Ru-NO) and at the nitrosylsulphito in 1 (Ru-N(O)SO3). The pKa value of the -SO3 moiety of the N(O)SO3 ligand in 1 was determined to be 5.08 ± 0.06 (at 298 K). The unprecedented photochemistry of a nitrosylsulphito complex is investigated in detail with 1. The proposed mechanism is based on experimental (UV-Vis, EPR, NMR and Transient Absorption Laser Flash Photolysis) and theoretical data (DFT) and involves photorelease of the N(O)SO3- ligand followed by formation of nitric oxide (NO˙) and sulfite radicals (SO3˙-, sulfur trioxide anion radical).

Mate extract as feed additive for improvement of beef quality.

Mate (Ilex paraguariensis A.St.-Hil.) is generally recognized as safe (GRAS status) and has a high content of alkaloids, saponins, and phenolic acids. Addition of mate extract to broilers feed has been shown to increase the oxidative stability of chicken meat, however, its effect on beef quality from animals supplemented with mate extract has not been investigated so far. Addition of extract of mate to a standard maize/soy feed at a level of 0.5, 1.0 or 1.5% w/w to the diet of feedlot for cattle resulted in increased levels of inosine monophosphate, creatine and carnosine in the fresh meat. The content of total conjugated linoleic acid increased in the meat as mate extract concentration was increased in the feed. The tendency to radical formation in meat slurries as quantified by EPR spin-trapping decreased as increasing mate extract addition to feed, especially after storage of the meat, indicating higher oxidative stability. Mate supplementation in the diet did not affect animal performance and carcass characteristics, but meat from these animals was more tender and consequently more accepted by consumers. Mate extract is shown to be a promising additive to feedlot diets for cattle to improve the oxidative stability, nutritive value and sensory quality of beef.

Photoinduced Charge Shifts and Electron Transfer in Viologen-Tetraphenylborate Complexes: Push-Pull Character of the Exciplex.

Viologen-tetraarylborate ion-pair complexes were prepared and investigated by steady-state and time-resolved spectroscopic techniques such as fluorescence and femtosecond transient absorption. The results highlight a charge transfer transition that leads to changes in the viologen structure in the excited singlet state. Femtosecond transient absorption reveals the formation of excited-state absorption and stimulated emission bands assigned to the planar (kobs < 1012 s-1) and twisted (kobs ∼ 1010 s-1) structures between two pyridinium groups in the viologen ion. An efficient photoinduced electron transfer from the tetraphenylborate anionic moiety to the viologen dication was observed less than 1 µs after excitation. This is a consequence of the push-pull character of the electron donor twisted viologen structure, which helps formation of the borate triplet state. The borate triplet state is deactivated further via a second electron transfer process, generating viologen cation radical (V•+).

New insight into the photophysics and reactivity of trigonal and tetrahedral arylboron compounds.

The photophysics and reactivity of two tetraphenylborate salts and triphenylborane have been studied using ultrafast transient absorption, steady-state fluorescence, electron paramagnetic resonance with spin trapping, and DFT calculations. The singlet excited state of tetraarylborates exhibit extended π-orbital coupling between two adjacent aryl groups. The maximum fluorescence band, as well as the transient absorption bands centered at 560 nm (τ = 1.05 ns) and 680 nm (τ = 4.35 ns) are influenced by solvent viscosity and polarity, indicative of a twisted intramolecular charge transfer (TICT) state. Orbital contour plots of the HOMO and LUMO orbitals of the tetraarylboron compounds support the existence of electron delocalization between two aryl groups in the LUMO. This TICT-state and aryl-aryl electron extension is not observed for the trigonal arylboron compound, in which excited π-orbital coupling only occurs between the boron atom and one aryl group, which restricts the twist motion of the aryl-boron bond. The excited triplet state is deactivated primarily through aryl-boron bond cleavage, yielding aryl and diphenylboryl radicals. In the presence of oxygen, this photochemistry results in phenoxyl and diphenylboroxyl radicals, as confirmed by EPR spectroscopy of spin trapped radical adducts. The TICT transition and radical generation is not expected for BoDIPY molecules where the rotational vibration of the B-aryl bond is rigid, restricting changes in the geometric structure. In this sense, this work contributes to the development of new BoDIPY derivatives where the TICT transition may be observed for aryl ligands with free rotational vibrations in the BoDIPY structure.

Limited proteolysis of myoglobin opens channel in ferrochelatase-globin complex for iron to zinc transmetallation.

Recombinant ferrochelatase (BsFECH) from Bacillus subtilis expressed in Escherichia coli BL21(DE3) was found by UV-visible spectroscopy to bind the model substrate tetraphenylporphyrin-sulfonate, TPPS, with Ka=3.8 10(5)mol/L in aqueous phosphate buffer pH 5.7 at 30°C, and to interact with metmyoglobin with Ka=1.07±0.13 10(5)mol/L at 30°C. The iron/zinc exchange in myoglobin occurring during maturation of Parma hams seems to depend on such substrate binding to BsFECH and was facilitated by limited pepsin proteolysis of myoglobin to open a reaction channel for metal exchange still with BsFECH associated to globin. BsFECH increased rate of zinc insertion in TPPS significantly and showed saturation kinetics with an apparent binding constant of Zn(II) to the [enzyme-TPPS] complex of 1.3 10(4)mol/L and a first-order rate constant of 6.6 10(-1)s(-1) for dissociation of the tertiary complex, a similar pattern was found for zinc/iron transmetallation in myoglobin.

Isomerization of Cholecalciferol through Energy Transfer as a Protective Mechanism Against Flavin-Sensitized Photooxidation.

Cholecalciferol, vitamin D3, was found to isomerize to 5,6-trans-vitamin-D3 with a quantum yield of 0.15 ± 0.01 in air-saturated 7/3 tert-butyl alcohol/water (v/v) at 25 °C, increasing to 0.32 ± 0.02 in the absence of oxygen, through quenching of triplet excited state flavin mononucleotide, FMN, rather than becoming oxidized. The quenching was found by laser flash photolysis to have a rate constant of 1.4 × 10(8) L mol(-1) s(-1) in 7/3 tert-butyl alcohol/water (v/v) at 25 °C, assigned to energy transfer from (3)FMN* to form a reactive vit.D3 diradical. vit.D3 forms a 1/1 precomplex with FMN by hydrophobic stacking with ΔH° = -36 ± 7 kJ mol(-1) and ΔS° = -4 ± 3 J mol(-1) K(-1), as shown by single photon counting fluorescence spectroscopy and steady-state fluorescence spectroscopy. Both ground-state precomplex formation and excited-state energy transfer seem important for vit.D3 protection against flavin-sensitized photooxidation of nutrients in food and biological systems.

Kinetics and thermodynamics of 1-hydroxyethyl radical reaction with unsaturated lipids and prenylflavonoids.

Hydroxyalkyl radicals have been reported to induce lipid oxidation as the key aspect of the pathogenesis of alcoholic fatty liver disease and are responsible for the alkylation and cleavage of DNA during the metabolism of a wide range of genotoxic compounds. However, relevant kinetic data for the oxidation of unsaturated lipids by 1-hydroxyethyl radical (HER) has not been reported. In this study, the rate constants for the reaction of unsaturated fatty acid methyl esters and sterols with HER have been determined using a competitive kinetic approach employing the spin-trap α-(4-pyridyl-1-oxide)-N-tert-butylnitrone (4-POBN) as the competitive substrate. Polyunsaturated fatty acid methyl ester is shown to react with HER with an apparent second-order rate constant ranging from (3.7 ± 0.1) × 10(6) L mol(-1) s(-1) for methyl linoleate to (2.7 ± 0.2) × 10(7) L mol(-1) s(-1) for methyl docosahexanoate at 25.0 ± 0.2 °C in ethanol. The apparent second-order rate constant for polyunsaturated fatty acid methyl ester oxidation by HER is dependent on the number of bisallylic hydrogen atoms rather than on the bond dissociation energy value for the weakest C-H bond as determined by ab initio density functional theory calculations. Sterols displayed higher reactivity compared to unsaturated fatty acid methyl esters with apparent second-order rate constants of (2.7 ± 0.1) × 10(6) and (5.2 ± 0.1) × 10(7) L mol(-1) s(-1) at 25.0 ± 0.2 °C in ethanol for cholesterol and ergosterol, respectively. Similar experiments with prenylflavonoids as potential herbal chemopreventive agents for preventing alcoholic liver diseases yield apparent second-order rate constants close to the diffusion control with kapp values of (1.5 ± 0.2) and (3.6 ± 0.1) × 10(9) L mol(-1)s(-1) for 6-prenylnarigerin and xanthohumol at 25.0 ± 0.2 °C in ethanol solution, respectively. Polyunsaturated lipids were revealed to be highly reactive oxidizable substrates toward HER-induced oxidation in biological systems leading to damage of membranes and sensitive structures.

Oxidation of carbon monoxide by perferrylmyoglobin.

Perferrylmyoglobin is found to oxidize CO in aerobic aqueous solution to CO2. Tryptophan hydroperoxide in the presence of tetra(4-sulfonatophenyl)-porphyrinate-iron(III) or simple iron(II)/(III) salts shows similar reactivity against CO. The oxidation of CO is for tryptophan hydroperoxide concluded to depend on the formation of alkoxyl radicals by reductive cleavage by iron(II) or on the formation of peroxyl radicals by oxidative cleavage by iron(III). During oxidation of CO, the tryptophan peroxyl radical was depleted with a rate constant of 0.26 ± 0.01 s(-1) for CO-saturated aqueous solution of pH 7.4 at 25 °C without concomitant reduction of the iron(IV) center. Carbon monoxide is as a natural metabolite accordingly capable of scavenging tryptophan radicals in myoglobin activated by peroxides with a second-order rate constant of (3.3 ± 0.6) × 10(2) L mol(-1) s(-1), a reaction that might be of importance in cellular membranes of the intestine for protection of tissue against radical damage during meat digestion.

Riboflavin photosensitized oxidation of myoglobin.

The reaction of the fresh meat pigment oxymyoglobin, MbFe(II)O2, and its oxidized form metmyoglobin, MbFe(III), with triplet-state riboflavin involves the pigment protein, which is oxidatively cleaved or dimerized as shown by SDS-PAGE and Western blotting. The overall rate constant for oxidation of MbFe(II)O2 by ³Rib is (3.0 ± 0.5) × 109 L·mol⁻¹·s⁻¹ and (3.1 ± 0.4) × 109 L·mol⁻¹·s⁻¹ for MbFe(III) in phosphate buffer of pH 7.4 at 25 °C as determined by laser flash photolysis. The high rates are rationalized by ground state hydrophobic interactions as detected as static quenching of fluorescence from singlet-excited state riboflavin by myoglobins using time-resolved fluorescence spectroscopy and a Stern-Volmer approach. Binding of riboflavin to MbFe(III) has K(a) = (1.2 ± 0.2) × 104 mol·L⁻¹ with ΔH° = -112 ± 22 kJ·mol⁻¹ and ΔS° = -296 ± 75 J·mol⁻¹·K⁻¹. For meat, riboflavin is concluded to be a photosensitizer for protein oxidation but not for discoloration.

Beer thiol-containing compounds and redox stability: kinetic study of 1-hydroxyethyl radical scavenging ability.

The 1-hydroxyethyl radical is a central intermediate in oxidative reactions occurring in beer. The reactivity of thiol-containing compounds toward 1-hydroxyethyl radical was evaluated in beer model solutions using a competitive kinetic approach, employing the spin-trap 4-POBN as a probe and by using electron paramagnetic resonance to detect the generated 1-hydroxyethyl/4-POBN spin adduct. Thiol-containing compounds were highly reactive toward the 1-hydroxyethyl radical with apparent second-order rate constants close to the diffusion limit in water and ranging from 0.5 × 109 L mol⁻¹ s⁻¹ for the His-Cys-Lys-Phe-Trp-Trp peptide to 6.1 × 109 L mol⁻¹ s⁻¹ for the reduced lipid transfer protein 1 (LTP1) isolated from beer. The reactions gave rise to a moderate kinetic isotope effect (k(H)/k(D) = 2.3) suggesting that reduction of the 1-hydroxyethyl radical by thiol-containing compounds takes place by hydrogen atom abstraction from the RSH group rather than electron transfer. The content of reduced thiols in different beers was determined using a previously established method based on ThioGlo-1 as the thiol derivatization reagent and detection of the derivatized thiols by reverse-phase liquid chromatography coupled to a fluorescence detector. The total level of thiol in beer (oxidized and reduced) was determined after a reduction step employing 3,3',3″-phosphanetriyltripropanoic acid (TCEP) as the disulfide reductant. A good correlation among total protein and total thiol content in different beers was observed. The results suggest a similar ratio between reduced thiols and disulfides in all of the tested beers, which indicates a similar redox state.

Photooxidation of other B-vitamins as sensitized by riboflavin.

Pyridoxal phosphate (PLP) was found to deactivate triplet-excited riboflavin (Rib) in aqueous solution with a deactivation constant of 3.0 ± 0.1 × 10(8) L mol(-1) s(-1) at 25 °C. Likewise, PLP was found to quench the fluorescence emission of (1)Rib* with (1)kq = 1.0 ± 0.1 × 10(11) L mol(-1) s(-1) as determined by steady state fluorescence. The rather high quenching constant suggests the formation of a ground state complex, which was further confirmed by time-resolved fluorescence measurements to yield a (1)Rib* deactivation constant of 3.4 ± 0.4 × 10(10) L mol(-1) s(-1). Triplet quenching is assigned as one-electron transfer rather than hydrogen-atom transfer from PLP to (3)Rib*, as the reaction quantum yield, Φ = 0.82, is hardly influenced by solvent change from water to D2O, Φ = 0.78. Neither biotin nor niacin deactivates the singlet- or triplet-excited riboflavin as it is expected from their higher oxidation potentials E > 2 V vs NHE.

Reduction of ferrylmyoglobin by hydrogen sulfide. Kinetics in relation to meat greening.

The hypervalent meat pigment ferrylmyoglobin, MbFe(IV)═O, characteristic for oxidatively stressed meat and known to initiate protein cross-linking, was found to be reduced by hydrogen sulfide to yield sulfmyoglobin. Horse heart myoglobin, void of cysteine, was used to avoid possible interference from protein thiols. For aqueous solution, the reactions were found to be second-order, and an apparent acid catalysis could be quantitatively accounted for in terms of a fast reaction between protonated ferrylmyoglobin, MbFe(IV)═O,H(+), and hydrogen sulfide, H2S (k2 = (2.5 ± 0.1) × 10(6) L mol(-1) s(-1) for 25.0 °C, ionic strengh 0.067, dominating for pH < 4), and a slow reaction between MbFe(IV)═O and HS(-) (k2 = (1.0 ± 0.7) × 10(4) L mol(-1) s(-1) for 25.0 °C, ionic strengh 0.067, dominating for pH > 7). For meat pH, a reaction via the transition state {MbFe(IV)═O···H···HS}([symbol: see text]) contributed significantly, and this reaction appeared almost independent of temperature with an apparent energy of activation of 2.1 ± 0.7 kJ mol(-1) at pH 7.4, as a result of compensation among activation energies and temperature influence on pKa values explaining low temperature greening of meat.