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荧光法测定外阴阴道念珠菌病中酸性蛋白酶活性

Fluorometric determination of acid proteinase activity in vulvovaginal candidosis.

作者信息

Kiliç N, Kuştimur S, Arslan S, Aldemir H

机构信息

Department of Biochemistry, Gazi University, Faculty of Medicine, Ankara, Turkey.

出版信息

Mycoses. 1996 Sep-Oct;39(9-10):347-51. doi: 10.1111/j.1439-0507.1996.tb00151.x.

Abstract

Vulvovaginal candidosis is one of the most frequent disorders in obstetrics and gynaecology. Candida albicans is commonly considered to be the true vaginopathic agent. The secreted acid proteinase might be especially relevant in the pathogenesis of vulvovaginal candidosis. A fluorometric determination of acid proteinase activity of clinical C. albicans isolates was carried out during the present work using fluorescamine. L-Leucyl-L-alanine was included as an internal standard and the results were expressed as nmoles of leucylalanine equivalents h-1 per 2 x 10(4) cells. The 13 isolates were taken from non-diabetic, non-pregnant women aged 22-35 years with vulvovaginal candidosis. Candida albicans ATCC 44858 was used as a control. The proteinase activity in culture supernatants was detectable starting from the mid- to late- exponential phase of growth, peaked between 30 and 46 h, and then declined. The control had an activity of 2.72 nmol h-1 per 2 x 10(4) cells, whereas eight of the samples had a lower activity (1.05 nmol h-1 per 2 x 10(4) cells on average) and five of the samples had a higher activity (6.53 nmol h-1 per 2 x 10(4) cells on average). The fluorometric determination of acid proteinase activity was found to be more reproducible and sensitive than the previously used spectrophotometric determinations.

摘要

外阴阴道念珠菌病是妇产科最常见的疾病之一。白色念珠菌通常被认为是真正的阴道病原体。分泌的酸性蛋白酶可能在外阴阴道念珠菌病的发病机制中尤为重要。在本研究中,使用荧光胺对临床分离的白色念珠菌的酸性蛋白酶活性进行了荧光测定。以L-亮氨酰-L-丙氨酸作为内标,结果以每2×10⁴个细胞每小时亮氨酰丙氨酸当量的纳摩尔数表示。这13株分离株取自年龄在22至35岁、患有外阴阴道念珠菌病的非糖尿病、非妊娠女性。白色念珠菌ATCC 44858用作对照。培养上清液中的蛋白酶活性从生长的指数中期到后期开始可检测到,在30至46小时之间达到峰值,然后下降。对照的活性为每2×10⁴个细胞2.72 nmol h⁻¹,而八个样品的活性较低(平均每2×10⁴个细胞1.05 nmol h⁻¹),五个样品的活性较高(平均每2×10⁴个细胞6.53 nmol h⁻¹)。发现酸性蛋白酶活性的荧光测定比以前使用的分光光度法测定更具可重复性和敏感性。

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