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Biomedical subjects

W Schwab

Publications and source records attributed to W Schwab.

At least 37 records · Page 2Linked to original sources

3-Hydroxy-4,5-dimethyl-2(5H)-furanone (Sotolon) causing an off-flavor: elucidation of its formation pathways during storage of citrus soft drinks.

Gas chromatography/olfactometry (GCO) and gas chromatography-mass spectrometry (HRGC-MS) revealed 3-hydroxy-4, 5-dimethyl-2(5H)-furanone (sotolon) to be responsible for the "burnt" and "spicy" off-flavor observed in citrus soft drinks during storage. Among the ingredients of citrus soft drinks, ethanol and ascorbic acid were identified as the essential precursors of sotolon. Two formation pathways were postulated by studies using (2)H (D)- and (13)C-labeled ethanol and ascorbic acid; i.e., sotolon is formed from two molecules of ethanol and carbons 2 and 3 of ascorbic acid (pathway 1), or it is generated from one molecule of ethanol and carbons 3-6 of ascorbic acid (pathway 2).

Ascorbic Acid↗

Novel 1,3-dioxanes from apple juice and cider.

Extracts obtained by XAD solid-phase extraction of apple juice and cider were separated by liquid chromatography on silica gel. Several new 1,3-dioxanes including the known 2-methyl-4-pentyl-1,3-dioxane and 2-methyl-4-[2'(Z)-pentenyl]-1,3-dioxane, were identified in the nonpolar fractions by GC/MS analysis and confirmed by chemical synthesis. The enantioselective synthesis of the stereoisomers of the 1,3-dioxanes was performed using (R)- and (R,S)-octane-1,3-diol and (R)- and (R,S)-5(Z)-octene-1,3-diol as starting material. Comparison with the isolated products indicated that the natural products consisted of a mixture of (2S,4R) and (2R,4R) stereoisomers in the ratio of approximately 10:1, except for 1,3-dioxanes generated from acetone and 2-butanone. It is assumed that the 1, 3-dioxanes are chemically formed in the apples and cider from the natural apple ingredients (R)-octane-1,3-diol, (R)-5(Z)-octene-1, 3-diol, (3R,7R)- and (3R,7S)-octane-1,3,7-triol, and the appropriate aldehydes and ketones, which are produced either by the apples or by yeast during fermentation of the apple juice.

Beverages↗

Isolation, identification, and enantioselective synthesis of octane-1,3,7-triol: determination of its absolute configuration

Extracts obtained by solid-phase extraction from apples were separated by multilayer countercurrent chromatography. In the most polar fractions, the novel octane-1,3,7-triol was identified by 1H and 13C NMR as well as LC-MS and by comparison with the synthesized racemic reference compound. Resolution of the enantiomers was achieved after acetylation of the triol followed by GC separation. The enantioselective synthesis of the stereoisomers of octane-1,3, 7-triol was performed using the building blocks (R)- and (R, S)-butane-1,3-diol and (S)- and (R,S)-butane-1,2,4-triol. Comparison with the isolated products indicated that the natural compound consisted of a mixture of (3R,7S)- and (3R,7R)-octane-1,3,7-triol in a ratio of 2:3. Since the C3 chiral center is enantiomerically pure, the triol might be biogenetically related to the known antimicrobial (R)-(+)-octane-1,3-diol, the major volatile compound of some apple cultivars.

Journal Article↗

Ultrastructural identification of caveolae and immunocytochemical as well as biochemical detection of caveolin in chondrocytes.

Using fluorescence immunocytochemistry, transmission electron microscopy and Western blotting, we have shown that caveolae and caveolin are abundant on chondrocytes of different cartilaginous structures of newborn and adult rat knee joints. Caveolin was detected in chondrocytes of the outer layer of articular cartilage, in the fibrocartilage of the menisci, and in fibrocartilage-like cells at tendon and ligament insertions. Electron microscopical studies revealed caveolae-like invaginations along the plasmalemmal membrane of articular chondrocytes and fibrocartilage cells. Immunoblot analysis demonstrated caveolin in detergent-insoluble and soluble complexes isolated from cultured rat chondrocytes.

Age Factors↗

Identification and characterization of potential new therapeutic targets in inflammatory and autoimmune diseases.

The isoxazole derivative Leflunomide (HWA 486) is a novel immunoregulatory and anti-inflammatory drug. Affinity chromatography was used to purify and identify Leflunomide binding proteins, which might play a role as potential cellular targets in the molecular mode of action. The Leflunomide derivative A 0273 was covalently coupled to a Fractogel(R) matrix. This column was used to separate a cytosolic protein extract of the macrophage cell line RAW 264.7 by several selected and specific gradient elution steps. Proteins that were specifically eluted through the active metabolite of Leflunomide, A 1726, were identified by subsequent protein sequence analysis. This allowed us to specify 10 cytosolic proteins, which bind with high affinity to this matrix. Three of them, glyceraldehyde 3-phosphate dehydrogenase, pyruvate kinase and phosphoglycerate mutase belong to the second part of the glycolytic pathway. The binding specificity of these protein/drug interactions was further evaluated using BIAcore(R) analysis. Kd values of glyceraldehyde 3-phosphate dehydrogenase, pyruvate kinase and lactic dehydrogenase were similar to the Kd value of a known Leflunomide target protein, dihydroorotate dehydrogenase. In order to elucidate the features as well as the overall relevance of these results, cytosolic fractions of three additional cell lines MOLT-4, A20.2J, HeLa were compared using the same chromatographic protocol. The elution profiles as well as subsequent Western blot analyses confirmed the data obtained previously for the macrophage cell line RAW 264.7.

Amino Acid Sequence↗

Immunohistochemical distribution of connexin 43 in the cartilage of rats and mice.

Using fluorescence immunohistochemistry, the distribution of connexin 43 was examined in hyaline cartilage and in the perichondrium of mouse and rat knee joints. In addition, rat chondrocytes were shown to be coupled in dye transfer studies with Lucifer Yellow. Connexin 43 was detected between chondrocytes in the outer layer of knee joint cartilage, between chondrocytes of the growth plate and between fibrocartilage-like cells at tendon and ligament insertions and in the tendons and ligaments proper. However, in the hyaline cartilage of the hind limbs of mature rats, the degree of connexin 43 immunoreactivity was diminished. These data suggest a possible involvement of connexins in cartilage development.

Aging↗

The impact of true partnership between a university Level I trauma center and a community Level II trauma center on patient transfer practices.

OBJECTIVE: To examine the effect of a clinical and administrative partnership with an academic urban Level I trauma center on the patient transfer practices at a suburban/rural Level II center. METHODS: Data for 2 years before affiliation (PRE) abstracted from inpatient charts and the trauma registry were compared with that for 2 years after (POST). The following data were collected: number of, reason for, and destination and demographics of transfers. Chi(2) test and t test analyses were used; p < 0.05 defined significance; data are mean +/- SEM. RESULTS: Transfer rate increased from 4% PRE to 6.9% (p = 0.001) POST with no significant difference in age, Glasgow Coma Scale score, Injury Severity Score, or Revised Trauma Score. Repatriation occurred in 12.8% POST (none PRE). The current Level I facility accepted 1.8% of all transfers PRE and 36.4% POST (p = 0.0001). PRE/POST rates by reason are as follows: pediatric, 14.6%/9.0% (p = 0.04); intensive care unit, 0.4%/1.7% (p = 0.13); complex orthopedic, 100%/0% (p = 0.005); vascular, 50%/0% (p = 0.008); spinal cord injury, 100%/100%; and ophthalmologic, 0%/100% (p = 0.005). CONCLUSIONS: In this experience of Level I/II partnership (1) transfer patterns were altered, (2) select patient cohort transfers decreased (pediatric, complex orthopedic, vascular), whereas others increased (aortic work-up), and (3) repatriation rates were low.

Hospitals, Community↗

Innervation pattern of different cartilaginous tissues in the rat.

BACKGROUND: The innervation of skeletal tissues by sensory nerves is poorly understood - especially of nerve fibres which reach into the bony and cartilaginous tissue. METHODS: Samples of rat cartilaginous tissues from different locations (knee joint, vertebral column, temporomandibular joint) were fixed by perfusion and decalcified. The distribution of protein gene product (PGP) 9.5-, calcitonin gene-related peptide (CGRP)- and tachykinin (TK)-immunoreactive axons was analysed using fluorescence immunohistochemistry. RESULTS: Nerve fibres were detected in the outer regions of the hyaline cartilage of the knee joint, in the hyaline cartilage of the vertebral body, in the fibrocartilage of the intervertebral disc and menisci, and in the articular disc of the temporomandibular joint. Predominantly, they were found to be CGRP-immunoreactive. CONCLUSION: The neuropeptidergic innervation of the hyaline cartilage in different locations and the presence of nerve fibres in the fibrocartilage might indicate that in addition to the classical neuronal afferent and efferent pathway these fibres may also mediate trophic actions like tissue adaptation and repair.

Animals↗

Microtopography of the autonomic nerves in the rat knee: a fluorescence microscopic study.

BACKGROUND: The autonomic innervation of the joint is involved in different functions, such as sensory inputs, modulation of the function of immune cells, and trophic actions. To have a basis for further studies of the arthritic knee joint we have investigated the topographical distribution of different neuropeptides in knees of newborn and adult rats and in adult rats after arthritis induction. METHODS: The distribution of the neuropeptides calcitonin gene-related peptide (CGRP), neurokinin A (NKA), substance P (SP), and neuropeptide Y (NPY) was analyzed using fluorescence immunohistochemistry. Samples were investigated after fixation by perfusion and decalcification by a special method which allows studies in bone tissue. Vascular structures were analyzed by scanning electron microscopy (SEM) of vascular resin casts. RESULTS: In all tissues of the joint (synovial membrane, vessels, fibrous structures, bone, and cartilagineous tissues) CGRP and NKA are the most frequent neuropeptides. They are localized in free or perivascular fibers predominantly around arteries and arterioles. The NPY-ergic perivascular fibers even enter the vessel wall. Generally, SP-ergic fibers occur rarely. Free NKA- and CGRP-ergic nerve fibers spread out in the synovial lining layer reaching the synovial cavity and the outer layers of the articular and metaphyseal cartilage. In the cartilagineous tissue these nerves contact the chondrocytes. The density of NKA- and CGRP-immunoreactive fibers is lower in newborn rats than in adult rats. Six hours after arthritis induction SP-, NKA-, and CGRP-immunoreactivity is enhanced especially in perivascular fibers. The related vessels are dilated substantially. CONCLUSIONS: The distribution pattern of the autonomic nerves found in this study might reflect the functions of these nerves: control of the microcirculation, sensory and even trophic functions. The new finding of CGRP- and NKA-ergic fibers in the outer layer of the cartilage can also have implications for the pathogenesis of rheumatoid arthritis.

Animals↗

Identification of 2,5-dimethyl-4-hydroxy-3[2H]-furanone beta-D-glucuronide as the major metabolite of a strawberry flavour constituent in humans.

2,5-Dimethyl-4-hydroxy-3[2H]furanone (Furaneol, DMHF) [3658-77-3], an important flavour constituent of strawberry fruit, was administered to four male and two female volunteers using fresh strawberries as a natural DMHF source. The amount excreted was determined by measuring urinary levels of DMHF and DMHF glucuronide. DMHF glucuronide was synthesized and the structure elucidated by mens of 1H, 13C and two dimensional nuclear magnetic resonance, as well as mass spectral data. Identification and quantification of DMHF glucuronide in human urine were achieved after solid phase extraction on XAD-2 using reverse-phase reverse-phase HPLC with either on-line UV/VIS or electrospray tandem mass spectrometry detection. Male and female volunteers excreted 59-69% and 81-94%, respectively, of the DMHF dose (total of free and glycosidically bound DMHF in strawberries) as DMHF glucuronide in urine within 24 hr. The amount of DMHF excretion was independent of the dose size and the ratio of free to glycosidically bound forms of DMHF in strawberry fruit. DMHF, DMHF glucoside and its 6'-O-malonyl derivative, naturally occurring in strawberries, were not detected in human urine.

Adult↗

Absolute configuration of octanol derivatives in apple fruits.

In extracts obtained by liquid-liquid extraction and enzymatic hydrolysis from five apple cultivars (Renao; Bedan; Peau de Chien; Noel des Champs; Red Delicious), chiral evaluation of free and glycosidically-bound octane-1,3-diol and 5(Z)-octene-1,3-diol, as well as ethyl 3-hydroxyoctanoate and ethyl 5(Z)-3-hydroxyoctenoate, was performed by multidimensional gas chromatography (MDGC), combining a polar achiral column (DB-Wax) with a chiral main column (2,3-di-O-acetyl-6-O-tert. butyldimethylsilyl-beta-cyclodextrin/OV 1701). Comparison of retention times of synthesized optically-enriched reference compounds with isolated diols and hydroxyesters, revealed the (R)-configuration for the free diols in cvs. Renao, Bedan, Peau de Chien and Noel des Champs and the (R)-configuration for the bound diols in cvs Bedan, Peau de Chien and Noel des Champs, exhibiting enantiomeric excesses (ees) greater than 99%. (R)-hydroxyesters (ee > 99%) were detected in cvs. Noel des Champs and Red Delicious.

Fruit↗

2,5-Dimethyl-4-hydroxy-3[2H]-furanone 6'O-malonyl-beta-D-glucopyranoside in strawberry fruits.

2,5-Dimethyl-4-hydroxy-3[2H]-furanone 6'-O-malonyl-beta-D-glucopyranoside was isolated from a glycosidic extract of strawberry fruit (Fragaria x ananassa, cv. Senga Sengana) by means of countercurrent chromatography and reverse-phase HPLC. Identification was achieved by comparison of chromatographic and 1H, 13C and 2D-NMR, as well as mass spectral data with those of the synthesized reference compound. In ripe strawberry fruit, a 1:1 ratio of the malonylated glucoside to its deacylated glucoconjugate was determined by on-line liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry.

Chromatography, High Pressure Liquid↗

Nuclear fragmentation of high-energy heavy-ion beams in water.

As a part of the physical-technical program of the heavy-ion therapy project at GSI we have investigated the nuclear fragmentation of high-energy ion beams delivered by the heavy-ion synchrotron SIS, using water as a tissue-equivalent target. For a direct comparison of fragmentation properties, beams of 10B, 12C, 14N, and 16O were produced simultaneously as secondary beams from a primary 18O beam and separated in flight by magnetic beam analysis. The Z-distributions of beam fragments produced in the water target were measured via energy loss in a large ionisation chamber and a scintillator telescope. From these data we obtained both total and partial charge-changing cross sections. In addition we have performed Bragg measurements using two parallel-plate ionization chambers and a water target of variable length. The detailed shape of the measured Bragg curves and the measured cross sections are in good agreement with model calculations based on semi-empirical formulae.

Boron↗