Aus Protoplasten von Candidu lipolytiea, Stamm 7 -3, wurden nach osmotisohem Schock dnrch Differentialzentrifugation bei 1500, 8000 und 16 000 g subzellulare Partikeln gewonnen. D i e Fraktionen katalysierten mit NADH als Cosubstrat den enzymatischen Abbau von n-Hexadecan-l-14C. Bei Substratkonzentrationen von 0,46 bis 1,05 pmol/ml betrugen der Abbau 20 bis 75,8% und die Umsatzrate 3,5 bis 17,9 ymolmg Pr0tein-l-h1. Die Reaktion war bereits nach 10 bis 15 min beendet. Die lipidloslichen Oxydationsprodukte wurden siinlenchrornatographisch vom Restalkan abgetrennt. In der wiilrigen Phase des Inkubationsansatzes konnten hydrophile Reektionsprodukta naohgewiesen werden. AuBerdem d e n die von den subzelluliiren Pastikeln absorbierten radioaktiv markierten Substanzen gemessen.
Various changes in the skin at menopause are based on "so-called dry skin". However, there is no evidence to explain the mechanism. Recently we found decreased hydration, impaired permeability barrier recovery and weakened integrity of the stratum corneum (SC) in an ovariectomized (OVX) mice model of climacterium. These changes in OVX-mice were restored by hormone replacement treatment (HRT) with 17beta-estradiol. The present study was designed to determine whether the dysfunction of the SC in OVX-mice was associated with changes of epidermal differentiation and corneodesmosome. Function of the SC was evaluated in HR mice at 6 weeks after ovariectomy or sham operation with or without HRT. Immediate after functional analysis, skin was taken for further analysis. Thickness of epidermis and dermis were comparable between OVX-and control-mice on HE-section, although those in HRT mice were thicker than others. Immunohistochemistry showed decreased expression of desmoglein-1, KLK7, loricrin, involucrin in OVX-mice compared with those in control-and HRT-mice. Real-time PCR showed increased levels of mRNA of desmoglein-1, KLK7, and SPINK5 in mice with HRT compared with other groups of mice, and those of KLK5 and corneodesmosin were comparable. In conclusion, reduced levels of hydration and impaired permeability barrier function of the SC in OVX-mice might be caused by impaired epidermal differentiation, and weakened integrity of the SC could be related to decreased levels of desmoglein-1, which might be associated with degradation of corneodesmosome. Thus, dysfunction of the SC found in OVX-mice model of climacterium should be a cause of symptom and a target for treatment at menopause.
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