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PubMed1997

Tyrosine kinase inhibitor-sensitive contractile action of ethanol in gastric smooth muscle: comparison with the action of epidermal growth factor.

We have evaluated the signal transduction pathways whereby, in comparison with epidermal growth factor-urogastrone, ethanol causes a rapid contractile response in guinea pig gastric longitudinal muscle. As for epidermal growth factor (EGF), the ethanol-induced contraction required extracellular calcium, was sensitive to the tyrosine kinase inhibitors genistein and tyrphostin 47 (AG213), and was blocked by both the cyclo-oxygenase inhibitor, indomethacin, and the diacylglycerol lipase inhibitor, U57908. The 50% effective concentration (EC50) for the contractile action of ethanol (approximately 140 mM) was lower than that for propanol and methanol and was not affected by the aldehyde dehydrogenase inhibitor, 4-methyl pyrazole. The actions of ethanol were distinct from those of EGF in that EGF-induced contractions were sensitive to the kinase C inhibitor GF109203X, and the EGF receptor kinase inhibitor PD153035, whereas ethanol-induced contractions were refractory to these inhibitors. Further, EGF-induced contractions were attenuated by the voltage-sensitive calcium channel antagonist, nifedipine, whereas the ethanol-induced contractile response was resistant to nifedipine but blocked by the "receptor-operated" calcium channel antagonist SKF96365. We conclude that ethanol without metabolism via alcohol dehydrogenase causes a contractile response in gastric longitudinal muscle tissue via a tyrosine kinase inhibitor-sensitive signal pathway that is parallel in many respects but yet is distinct from that activated by EGF.

1-Propanol↗
PubMed1997

Use of a fluorescent analog of CDP-DAG in human skin fibroblasts: characterization of metabolism, distribution, and application to studies of phosphatidylinositol turnover.

We studied the uptake metabolism, and distribution of a fluorescent analog of CDP-diacylglycerol [cytidine diphosphate-1, 2-oleoyl, (N-(4-nitrobenzo-2-oxa-1,3-diazole) aminocaproyl) diacylglycerol; CDP-NBD-DAG]. When cells were incubated with CDP-NBD-DAG for 60 min at 11 degrees C and washed, the fluorescent lipid was localized to the plasma membrane. However, upon warming to 37 degrees C, the fluorescent lipid redistributed into various intracellular membranes and was metabolized primarily to fluorescent analogs of DAG and phosphatidylcholine (PC), although small amounts of fluorescent phosphatidic acid and phosphatidylinositol (PI) were also formed. The incorporation of 32Pi into some of the fluorescent lipids was also determined in order to assess their turnover. Stimulation of cells with platelet-derived growth factor enhanced the synthesis of fluorescent PI relative to unstimulated cells by approximately 68%, while the synthesis of fluorescent PC was unaffected. In addition, the incorporation of 32Pi into fluorescent PI was enhanced. Stimulation of cells with interleukin-1 beta enhanced the synthesis of both fluorescent PI (approximately 88%) and PC (approximately 250%) compared to non-stimulated cells, but with less incorporation of 32Pi into fluorescent PI. Finally, incubation of CDP-NBD-DAG-treated cells with inhibitors of phosphatidic acid phosphohydrolase and DAG kinase resulted in a dramatic increase in the amount of fluorescent PI formed (approximately 64% of all the CDP-NBD-DAG metabolites). We conclude that CDP-NBD-DAG can be used for the de novo synthesis of fluorescent PI, and in combination with 32P labeling, provides a convenient method for studying PI turnover.

4-Chloro-7-nitrobenzofurazan↗
PubMed1996

[Correlative study on the storage time and quality of rhizoma Dryopteris crassirhizomae].

The contents of dryocrassin in Rhizoma Dryopteris Crassirhizomae were determined by TLC-scanning in a period of two years of storage. Phloroglucinol content was calculated by area normalization method. The results show that in contrast with the fresh crude drugs of Rhizoma Dryopteris Crassirhizomae, the contents of dryocrassin in the crude drugs of one year and two years of storage have reduced by 22% and 27% respectively, and phloroglucinol contents have reduced by 2% and 8% respectively.

Benzylidene Compounds↗
PubMed1997

Contractile action of ethanol in guinea pig gastric smooth muscle: inhibition by tyrosine kinase inhibitors and comparison with the contractile action of epidermal growth factor-urogastrone.

We observed a contractile action of ethanol (20-500 mM) and other alcohols (methanol and propanol, but not butanol) in guinea pig gastric longitudinal (LM) and circular (CM) smooth muscle preparations. The potency order for the alcohols in the LM preparation was: ethanol = propanol > methanol; and in the CM preparation, propanol > ethanol > methanol. Like epidermal growth factor-urogastrone (EGF), the contractile actions of ethanol in the LM and CM preparations required extracellular calcium and were blocked by the tyrosine kinase inhibitors, genistein and tyrphostin-47 (AG213). The tyrosine phosphatase inhibitor, pervanadate, potentiated the contractile action of ethanol in the LM preparation. Ethanol-induced contractions in both preparations were not affected by 4-methyl pyrazole, an inhibitor of alcohol dehydrogenase, and were unaffected by tetrodotoxin, atropine, prazosine or yohimbine. In the LM preparation, like EGF, the contractile action of ethanol was blocked by the cyclooxygenase inhibitor, indomethacin, and the diacylglycerol lipase inhibitor, U57,908; in the CM preparation, contractions caused by ethanol and EGF were still observed in the presence of these two inhibitors. Contractions caused by ethanol and EGF in the LM preparation were not affected by the epoxygenase inhibitor, ketoconazole; the lipoxygenase inhibitor, nordihydroguaiaretic acid; or the phospholipase A2 inhibitor, mepacrine. In contrast, in the LM preparation, EGF-induced contractions were attentuated by the EGF receptor-kinase inhibitor, PD153035; the MAP-kinase-kinase (MEK) inhibitor, PD98059; the kinase C inhibitor, GF109203X; and the phosphatidylinositol 3'-kinase inhibitors, Wortmannin and LY294002; whereas ethanol-induced contractions were unaffected by these inhibitors. Both ethanol and EGF caused small increases in the phosphotyrosyl protein content of the gastric tissue. We conclude that ethanol causes its contractile effects in the distinct gastric LM and CM preparations independent of nerve-released agonists and via a tyrosine kinase inhibitor-sensitive signal pathway that is in many respects similar to, but distinct from the one activated by EGF.

Animals↗
PubMed1997

Allergic contact dermatitis from a wooden necklace.

A 36-year-old female kitchenworker twice developed eczematous lesions corresponding exactly to the area around her neck where she had worn a wooden necklace. Contact dermatitis lasted longer than 1 week. The necklace consisted of 42 brown wooden beads and 63 other wooden parts, 0.5 to 3 cm diameter. Most parts could be identified as Cocobolo wood, Brazilian and East Indian rosewood, and teak. Patch tests with the pure constituents gave +3-reactions to three dalbergions and obtusaquinone, which are known to be the sensitizers of Cocobolo and the above-mentioned rosewoods. Because of these test results, the identification of the species by eye examination could be corroborated. Further detailed questioning revealed that the patient had played a recorder, probably made from Cocobolo (Dalbergia retusa), when a child, to which she unknowingly became allergic.

Adult↗
PubMed1997

[Late onset type I tyrosinemia].

The authors present a case of tyrosinemia type 1, 3 years old girl at the time of diagnosis. The presenting symptoms were 3 times colic, obstipation, acute encephalopathy, hypertension, hyponatremia, according to the porphyric crisis. Her kidney function tests gave normal results during illness, only once an increased calcium turnover was observed. She has no singe of rachitis. Cirrhosis of the liver was proved by biopsy because of progressively rising gammaGT and alfa-fetoprotein levels. A new ensime-blocker (NTBC) treatment was started in an international collaboration. The authors compare the history of this case to that of others published in the literature. They summarize the pathomechanism of the disease.

Amino Acid Metabolism, Inborn Errors↗
PubMed1998

Neurite outgrowth stimulated by neural cell adhesion molecules requires growth-associated protein-43 (GAP-43) function and is associated with GAP-43 phosphorylation in growth cones.

The mechanisms whereby cell adhesion molecules (CAMs) promote axonal growth and synaptic plasticity are poorly understood. Here we show that the neurite outgrowth stimulated by NCAM-mediated fibroblast growth factor (FGF) receptor activation in cerebellar granule cells is associated with increased GAP-43 phosphorylation on serine-41. In contrast, neither NCAM nor FGF was able to stimulate neurite outgrowth in similar neurons from mice in which the GAP-43 gene had been deleted by homologous recombination. Integrin-mediated neurite outgrowth was unaffected by GAP-43 deletion. Both neurite outgrowth and rapid phosphorylation of GAP-43 in isolated growth cones required the first three Ig domains of a NCAM-Fc chimera and were stimulated maximally at 5 micrograms/ml (approximately 50 nM). Likewise, GAP-43 phosphorylation in isolated growth cones also was stimulated by an L1-Fc chimera. Both neurite outgrowth and NCAM-stimulated GAP-43 phosphorylation were inhibited by antibodies to the FGF receptor and a diacylglycerol lipase inhibitor (RHC80267) that blocks the production of arachidonic acid in response to activation of the FGF receptor. Direct activation of the FGF receptor and the arachidonic acid cascade with either basic FGF or melittin also resulted in increased GAP-43 phosphorylation. These data suggest that the stimulation of neurite outgrowth by NCAM requires GAP-43 function and that GAP-43 phosphorylation in isolated growth cones occurs via an FGF receptor-dependent increase in arachidonic acid.

3T3 Cells↗
PubMed1998

Vibrational circular dichroism: an incisive tool for stereochemical applications.

Vibrational circular dichroism (VCD) is the extension of circular dichroism (CD) from electronic to vibrational transitions in molecules. In this review, we present the basic concepts of VCD and focus on its growing strength for stereochemical applications. There are three principal areas of application of VCD. These are the determination of optical purity, the determination of absolute configuration and the determination of conformational properties of chiral molecules in solution. Until recently, the application of VCD was restricted to the use of instruments that had been custom assembled in academic research laboratories. This situation has now changed. It is possible to purchase a dedicated Fourier transform infrared (FTIR) VCD spectrometer complete with user friendly software. Combined with rapid advances in the ab initio theoretical prediction of infrared (IR) and VCD spectra, the availability of such technology opens the way for the widespread use of VCD spectroscopy for research and industrial applications.

Camphor↗