Qualitative and Quantitative Profiling of Fructose Degradation Products Revealed the Formation of Thirteen Reactive Carbonyl Compounds and Higher Reactivity Compared to Glucose.

Fructose occurs in foods and as a metabolite in vivo. It can be degraded, leading to the formation of reactive carbonyl compounds, which may influence food properties and have an impact on health. The present study performed an in-depth qualitative and quantitative profiling of fructose degradation products. Thus, the α-dicarbonyl compounds 3-deoxyglucosone, glucosone, methylglyoxal, glyoxal, hydroxypyruvaldehyde, threosone, 3-deoxythreosone, and 1-desoxypentosone and the monocarbonyl compounds formaldehyde, acetaldehyde, glycolaldehyde, glyceraldehyde, and dihydroxyacetone were detected in fructose solutions incubated at 37 °C. Quantitative profiling after 7 days revealed 4.6-271.6-fold higher yields of all degradation products from fructose compared to glucose. Except for 3-deoxyglucosone, the product formation appeared to be metal dependent, indicating oxidative pathways. CaCl2 and MgCl2 partially reduced fructose degradation. Due to its high reactivity compared to glucose, particularly toward metal-catalyzed pathways, fructose may be a strong contributor to sugar degradation and Maillard reaction in foods and in vivo.

Dried Vegetables as Potential Clean-Label Phosphate Substitutes in Cooked Sausage Meat.

While phosphates are key additives in sausage production, their use conflicts with consumer preferences for "natural" foods. In this study, we investigated the potential of using vegetables as "clean-label" phosphate substitutes and their effects on water holding capacity, consumer acceptance, color, softness, and tenderness. Six freeze-dried vegetables with a pH above 6.0 were added to sausage meat on a laboratory scale. Adding 1.6% freeze-dried Brussels sprouts or Red Kuri squash resulted in a similar weight gain (7.0%) as the positive control of 0.6% commercial phosphate additive. Higher vegetable concentrations (2.2-4.0%) caused a significant increase in weight (p ≤ 0.05, 10.4-18.4% weight gain). Similar stress was needed to compress sausages containing 1.6/4.0% Brussels sprouts (14.2/11.2 kPa) and the positive control (13.2 kPa). Indentation tests also led to similar softness results for the sausages prepared with 1.6/4.0% Brussels sprouts (15.5 kPa/16.6 kPa) and the positive control (16.5 kPa). A force of 1.25 N was needed to shear the positive control, while 1.60 N/1.30 N was needed for the samples (1.6/4% Brussels sprouts). In summary, the present study indicates that freeze-dried vegetables have the potential to effectively replace phosphate in meat products.

Degradation and de novo formation of nine major glucose degradation products during storage of peritoneal dialysis fluids.

Reactive glucose degradation products (GDPs) are formed during heat sterilization of glucose-containing peritoneal dialysis fluids (PDFs) and may induce adverse clinical effects. Long periods of storage and/or transport of PDFs before use may lead to de novo formation or degradation of GDPs. Therefore, the present study quantified the GDP profiles of single- and double-chamber PDFs during storage. Glucosone, 3-deoxyglucosone (3-DG), 3-deoxygalactosone (3-DGal), 3,4-dideoxyglucosone-3-ene (3,4-DGE), glyoxal, methylglyoxal (MGO), acetaldehyde, formaldehyde, and 5-hydroxymethylfurfural (5-HMF) were quantified by two validated UHPLC-DAD methods after derivatization with o-phenylenediamine (dicarbonyls) or 2,4-dinitrophenylhydrazine (monocarbonyls). The PDFs were stored at 50 °C for 0, 1, 2, 4, 13, and 26 weeks. The total GDP concentration of single-chamber PDFs did not change considerably during storage (496.6 ± 16.0 µM, 0 weeks; 519.1 ± 13.1 µM, 26 weeks), but individual GDPs were affected differently. 3-DG (- 82.6 µM) and 3-DGal (- 71.3 µM) were degraded, whereas 5-HMF (+ 161.7 µM), glyoxal (+ 32.2 µM), and formaldehyde (+ 12.4 µM) accumulated between 0 and 26 weeks. Acetaldehyde, glucosone, MGO, and 3,4-DGE showed time-dependent formation and degradation. The GDP concentrations in double-chamber fluids were generally lower and differently affected by storage. In conclusion, the changes of GDP concentrations during storage should be considered for the evaluation of clinical effects of PDFs.

Chemistry and clinical relevance of carbohydrate degradation in drugs.

Carbohydrate degradation products are formed during heat sterilization in drugs containing (poly-)glucose as osmotic agents. Given this situation, peritoneal dialysis fluids (PDFs) and infusion fluids are of particular clinical relevance, because these drugs deliver process contaminants either over a longer period or directly into the circulation of patients who are critically ill. For the development of suitable mitigation strategies, it is important to understand the reaction mechanisms of carbohydrate degradation during sterilization and how the resulting products interact with physiological targets at the molecular level. Furthermore, reliable, comprehensive, and highly sensitive quantification methods are required for product control and toxicological evaluation.

GABA(A) receptor modulation by terpenoids from Sideritis extracts.

SCOPE: GABAA receptors are modulated by Sideritis extracts. The aim of this study was to identify single substances from Sideritis extracts responsible for GABAA receptor modulation. METHODS AND RESULTS: Single volatile substances identified by GC have been tested in two expression systems, Xenopus oocytes and human embryonic kidney cells. Some of these substances, especially carvacrol, were highly potent on GABAA receptors composed of α1ß2 and α1ß2γ2 subunits. All effects measured were independent from the presence of the γ2 subunit. As Sideritis extracts contain a high amount of terpenes, 13 terpenes with similar structure elements were tested in the same way. Following a prescreening on α1ß2 GABAA receptors, a high-throughput method was used for identification of the most effective terpenoid substances on GABA-affinity of α1ß2γ2 receptors expressed in transfected cell lines. Isopulegol, pinocarveol, verbenol, and myrtenol were the most potent modifiers of GABAA receptor function. CONCLUSION: Comparing the chemical structures, the action of terpenes on GABAA receptors is most probably due to the presence of hydroxyl groups and a bicyclic character of the substances tested. We propose an allosteric modulation independent from the γ2 subunit and similar to the action of alcohols and anesthetics.

Endogenous mitochondrial oxidative stress in MnSOD-deficient mouse embryonic fibroblasts promotes mitochondrial DNA glycation.

The accumulation of somatic mutations in mitochondrial DNA (mtDNA) induced by reactive oxygen species (ROS) is regarded as a major contributor to aging and age-related degenerative diseases. ROS have also been shown to facilitate the formation of certain advanced glycation end-products (AGEs) in proteins and DNA and N(2)-carboxyethyl-2'-deoxyguanosine (CEdG) has been identified as a major DNA-bound AGE. Therefore, the influence of mitochondrial ROS on the glycation of mtDNA was investigated in primary embryonic fibroblasts derived from mutant mice (Sod2(-/+)) deficient in the mitochondrial antioxidant enzyme manganese superoxide dismutase. In Sod2(-/+) fibroblasts vs wild-type fibroblasts, the CEdG content of mtDNA was increased from 1.90 ± 1.39 to 17.14 ± 6.60 pg/µg DNA (p<0.001). On the other hand, the CEdG content of nuclear DNA did not differ between Sod2(+/+) and Sod2(-/+) cells. Similarly, cytosolic proteins did not show any difference in advanced glycation end-products or protein carbonyl contents between Sod2(+/+) and Sod2(-/+). Taken together, the data suggest that mitochondrial oxidative stress specifically promotes glycation of mtDNA and does not affect nuclear DNA or cytosolic proteins. Because DNA glycation can change DNA integrity and gene functions, glycation of mtDNA may play an important role in the decline of mitochondrial functions.

Identification of H2O2 as a major antimicrobial component in coffee.

Coffee shows distinct antimicrobial activity against several bacterial genera. The present study investigated molecular mechanisms and active ingredients mediating the antimicrobial effect of coffee. Depending on concentration, roasted, but not raw coffee brew inhibited the growth of Escherichia coli and Listeria innocua. Several coffee ingredients with known antibacterial properties were tested for their contribution to the observed effect. In natural concentration, caffeine, ferulic acid and a mixture of all test compounds showed very weak, but significant activity, whereas trigonelline, 5-(hydroxymethyl)furfural, chlorogenic acid, nicotinic acid, caffeic acid, and methylglyoxal were not active. Antimicrobial activity, however, was completely abolished by addition of catalase indicating that H(2)O(2) is a major antimicrobial coffee component. In accordance with this assumption, bacterial counts during 16 h of incubation were inversely related to the H(2)O(2) concentration in the incubation solution. Pure H(2)O(2) showed slightly weaker activity. The H(2)O(2) dependent antimicrobial activity of coffee could be mimicked by a reaction mixture of d-ribose and l-lysine (30 min 120 °C) indicating that H(2)O(2) is generated in the coffee brew by Maillard reaction products. Identification of H(2)O(2) as major antimicrobial coffee component is important to evaluate the application of coffee or coffee extracts as natural preservatives.

Immunochemical detection of tissue from the central nervous system via myelin proteolipid protein: adaptation for food inspection and development of recombinant bivalent Fab mini-antibodies.

Bovine spongiform encephalopathy (BSE) is transmitted by the ingestion of central nervous system (CNS) tissue of infected animals. Food inspection must, therefore, test for the presence of CNS tissue in meat and meat products. A Western blot assay for the specific CNS tissue marker myelin proteolipid protein (PLP) was optimized with considerably reduced analysis time, solvent consumption, and detection limit (0.001% CNS tissue in minced beef). Further, a PLP-specific recombinant bivalent fragment antigen binding mini-antibody (anti-PLP Fab) was obtained from a commercial phage display library. Western blot analysis with the anti-PLP Fab selectively detected CNS tissue in minced beef with a detection limit of 0.025%. Model experiments for meat processing revealed that assay sensitivity decreased with increasing temperature and prolonged heating time. A market survey with 687 sausage samples was performed using PLP-Western blot and enzyme-linked immunosorbent assay (ELISA) for glial fibrillary acidic protein (GFAP). Five samples were tested clearly positive by both assay systems, whereas in an additional six samples, CNS tissue was detected only by GFAP ELISA and in two samples only by PLP-Western blot.

Development and validation of an enzyme-linked immunosorbent assay (ELISA) for quantification of lysozyme in cheese.

A competitive enzyme-linked immunosorbent assay (ELISA) was developed to quantify the amount of the preservative and potential allergen lysozyme in cheese using a commercially available monoclonal antibody against hen egg white lysozyme. The limit of detection for lysozyme in a cheese matrix amounted to 2.73 ng/mL, and the working range comprises 3.125-800 ng/mL. Intra- and interassay coefficients of variation were lower than 12%. Neither cross-reactivity with alpha-lactalbumin and human lysozyme nor unspecific interference with matrix components was observed. The recovery of lysozyme-spiked cheese ranged from 87.4 to 93.6% at four concentrations (50, 100, 200, and 400 mg/kg). The ELISA method was also compared to a high-performance liquid chromatography (HPLC) method, confirming the reliability and accuracy of the ELISA. A total of 21 commercially available cheese samples produced with and without lysozyme were analyzed with ELISA as well as HPLC. Both methods showed good agreement with a correlation index of R2=0.990.