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PubMed1985

Purification and characterization of protease-resistant secretory granule proteoglycans containing chondroitin sulfate di-B and heparin-like glycosaminoglycans from rat basophilic leukemia cells.

Proteoglycans were extracted from nuclease-digested sonicates of 10(9) rat basophilic leukemia (RBL-1) cells by the addition of 0.1% Zwittergent 3-12 and 4 M guanidine hydrochloride and were purified by sequential CsCl density gradient ultracentrifugation, DE52 ion exchange chromatography, and Sepharose CL-6B gel filtration chromatography under dissociative conditions. Between 0.3 and 0.8 mg of purified proteoglycan was obtained from approximately 1 g initial dry weight of cells with a purification of 200-800-fold. The purified proteoglycans had a hydrodynamic size range of Mr 100,000-150,000 and were resistant to degradation by a molar excess of trypsin, alpha-chymotrypsin, Pronase, papain, chymopapain, collagenase, and elastase. Amino acid analysis of the peptide core revealed a preponderance of Gly (35.4%), Ser (22.5%), and Ala (9.5%). Approximately 70% of the glycosaminoglycan side chains of RBL-1 proteoglycans were digested by chondroitinase ABC and 27% were hydrolyzed by treatment with nitrous acid. Sephadex G-200 chromatography of glycosaminoglycans liberated from the intact molecule by beta-elimination demonstrated that both the nitrous acid-resistant (chondroitin sulfate) and the chondroitinase ABC-resistant (heparin/heparan sulfate) glycosaminoglycans were of approximately Mr 12,000. Analysis of the chondroitin sulfate disaccharides in different preparations by amino-cyano high performance liquid chromatography revealed that 9-29% were the unusual disulfated disaccharide chondroitin sulfate di-B (IdUA-2-SO4----GalNAc-4-SO4); the remainder were the monosulfated disaccharide GlcUA----GalNAc-4-SO4. Subpopulations of proteoglycans in one preparation were separated by anion exchange high performance liquid chromatography and were found to contain chondroitin sulfate glycosaminoglycans whose disulfated disaccharides ranged from 9-49%. However, no segregation of subpopulations without both chondroitin sulfate di-B and heparin/heparan sulfate glycosaminoglycans was achieved, suggesting that RBL-1 proteoglycans might be hybrids containing both classes of glycosaminoglycans. Sepharose CL-6B chromatography of RBL-1 proteoglycans digested with chondroitinase ABC revealed that less than 7% of the molecules in the digest chromatographed with the hydrodynamic size of undigested proteoglycans, suggesting that at most 7% of the proteoglycans lack chondroitin sulfate glycosaminoglycans.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗
PubMed1993

Positive- and negative-ion mass spectrometry of diphenylmethane antihistaminics and their analogues and rapid clean-up of them from biological samples.

Positive-ion electron impact (PIEI), positive-ion chemical ionization (PICI) and negative-ion chemical ionization (NICI) mass spectra are presented for 15 compounds of diphenylmethane antihistaminics and their analogues, and each fragmentation pathway was analyzed. In the PIEI mode, molecular peaks were very small or missing for most compounds. Peaks at m/z 58, due to a dimethylaminomethyl group liberated, constituted base peaks in five compounds. Peaks at m/z 165 and/or 167, due to diaromatic rings plus a methyl group, appeared in most compounds. In the PICI mode, peaks due to M+H and M+C2H5 appeared in all compounds. Peaks due to diaromatic rings plus a methyl or ethyl group constituted base peaks in five compounds, which had an ether bond in their structures. In the NICI mode, anions at m/z M-H appeared in most compounds. Peaks at m/z 35 were observed for compounds having a chlorine group in their structures. Detection limits for total ion monitoring of these compounds were 20-50 ng on column in the PIEI mode, 100-200 ng in the PICI mode and 500-1000 ng in the NICI mode. A rapid and simple clean-up procedure of these drugs with use of Sep-Pak C18 cartridges is also presented. The drugs could be detected by gas chromatography with DB-1 and DB-17 capillary columns with satisfactory separation from impurities in their underivatized forms. The recovery of the drugs, which had been added to whole blood and urine, was more than 60% except for meclizine.

Benzhydryl Compounds↗
PubMed1981

[Pharmacokinetics and efficacy of skin medications].

Topical therapy of the skin begins with the liberation from a carrier substance and results in a subsequent diffusion of the drug into the horny layer, depending on the physical and chemical properties of the respective agents. By permeating the epidermis the drug may be taken up by the inner compartment, namely the blood. A rediffusion to the skin is possible following systemic application. Although equivalent processes are induced by systemic and topical application, the latter application form limits the permeability including the first pass effect to the skin and its microcirculation. The characteristics are influenced by the type of dermatoses and by the condition of the horny layer as a barrier. The binding of these drugs to the skin's fibers and cells as well as the intensity and degree of permeation in the microcirculation is also of importance. The topical application of antihistamines, corticosteroids, heparin, coffein and nicotinic acid esters are used to describe the relationship between pharmacokinetics and effectiveness in view of clinical-experimental aspects. This pharmacoanalysis is based for the most part on factors which influence the microcirculation, namely the development of erythema, edema and thermography. The pharmacokinetics of topically applied substances are altered by the therapeutic effectivity of these agents and by the resulting changes in the structures of the skin.

Adrenal Cortex Hormones↗
PubMed1984

The relationship of inhaled sulfur dioxide reactivity to ingested metabisulfite sensitivity in patients with asthma.

When ingested in acid solution, the preservative sodium metabisulfite (MB) provokes asthma within minutes of ingestion in a proportion of asthmatic subjects. Freshly prepared acid solutions of MB liberate significant quantities of gaseous SO2. In order to test the hypothesis that asthma provoked by ingestion of acidified solutions of MB was due to supersensitivity to SO2 inhaled during swallowing, 3 groups of 10 subjects were studied. Groups 1 and 2 were asthmatics. Group 1 subjects were reactors and Group 2 were nonreactors to ingested MB. Group 3 subjects were nonasthmatic control subjects. Subjects were challenged, on separate days, with 50 mg of MB in citric acid and SO2 gas, via a steady-state system, at increasing concentrations (0.5, 1.5, 3, 5 ppm). The mean percent fall in peak expiratory flow rates after ingestion of MB for Group 1 was 35 +/- 14, Group 2 was 6 +/- 6, and Group 3 was 5 +/- 3. The mean SO2 provocation concentration (Pc20.SO2) for Group 1 was 1.19 +/- 0.78 ppm, for Group 2 it was 2.25 +/- 1.42 ppm, and for Group 3 it was greater than 5 ppm. The means for Pc20.SO2 of Groups 1 and 2 were not statistically different (p = 0.075), although the possibility of a Type 2 error is recognized. Five MB-sensitive subjects were subsequently challenged with the MB solution by mouthwash and nasogastric tube. Asthma was provoked in all 5 subjects by mouthwash but not by gastric challenge.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1995

[Intraoperative anaphylaxis to latex in pregnancy].

CASE REPORT: A 31-year-old pregnant woman had to undergo emergency abdominal surgery due to acute intestinal obstruction. The patient's preoperative history demonstrated multiple allergies as well as abdominal trauma several years before. The physical examination--including sonography of the two fetuses--showed no pathological signs. Anaesthesia was induced intravenously with the operating table tilted to the left side, using routine precautions pre-oxygenation, and rapid sequence intubation, and was maintained unproblematically. About 20 min after the onset of surgery, hypotension, tachycardia, and a drop in oxygen saturation appeared. Volume substitution and the application of vasoactive drugs failed to stabilise the haemodynamic situation. Elevation of the pregnant uterus and increased left tilzing of the operating table did not lead to improvement. The development of eyelid edema led to the diagnosis of an anaphylactic reaction. The patient was treated successfully with epinephrine, antihistamines, and corticosteroids (prednisolone). The suspicion of latex-related allergy was verified postoperatively by radio-allergen-sorbent test (RAST) and prick and scratch tests. DISCUSSION: The unspecific symptoms primarily led to the diagnosis of a prostacycline (liberated from the intestines)-induced, so-called eventration syndrome or aorto-caval compression syndrome, respectively, caused by the pregnant uterus [2, 7, 16, 17]. The initial therapeutic failure and the eyelid edema led to the correct diagnosis of an allergic reaction. Besides the application of epinephrine, which was indicated in spite of its vasoconstrictive effect on the smooth muscle of the uterus, immediate left-side-down positioning of the operating table, and sufficient volume replacement were decisive for haemodynamic stabilisation and maintenance of an adequate perfusion pressure of the uterus [7, 8, 13, 15]. Because of the non-specificity of RAST screening, the cutaneous tests had great significance in confirming the diagnosis of latex-related allergy [3, 18, 24]. CONCLUSIONS: Due to the obviously increasing number of latex-related allergies, especially in atopic persons and patients with frequent latex exposure, the patient's exact history is highly significant [4, 7, 18-20]. This includes--because of suspected cross-reactions--questions concerning allergic reactions to bananas and chestnuts [1, 16, 24]. If a latex-related allergy is suspected, all latex- or rubber-containing materials have to be consequently avoided. Because of the suspected allergies by inhalation via rubber-containing masks or tracheal tubes, these devices also have to be avoided and replaced, possibly by silicone materials [1, 4, 5, 16]. Premedication with H1- and H2-antagonists (dimetindene and cimetidine) and glucocorticoids (administered 12h before surgery and given twice) is indicated [5, 19, 12, 21]. In cases of latex allergy, the above-mentioned basic therapeutic measures have to be undertaken even in pregnancy, including immediate replacement of all latex-containing materials. The diagnosis of latex allergy should be verified by cutaneous testing [4, 18, 24].

Adult↗