Altered glomerular permeability as a result of focal detachment of the visceral epithelium.
Altered glomerular permeability as a result of focal detachment of the visceral epithelium.
复制标题
内脏上皮局部脱离导致肾小球通透性改变。
DOI：
10.1038/ki.1982.63
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发表时间：
1982
影响因子：
19.6
通讯作者：
Rosenzweig,LJ
中科院分区：
文献类型：
--
作者：
Kanwar,YS;Rosenzweig,LJ
方法使用体重为150 - 200 g的雄性Sprague-Dawley大鼠（Charles River Breeding Laboratories，North威尔明顿，马萨诸塞州）。材料购买如下：色谱纯化的神经氨酸酶（产气荚膜梭菌）和大豆胰蛋白酶抑制剂（SBTI），来自Worthington Bio-chemical Corporation，Freehold，新泽西;嘌呤霉素氨基糖苷（PAN）（6-二甲基氨基-9 [3 ′氨基-3 ′脱氧核糖基嘌呤]），购自Sigma Chemical Company，St. Louis，Missou-ri;来自Calbiochem，SanDiego，加州的马脾铁蛋白，两次结晶（无镉）;和来自ReheisChemicalCompany，Phoenix，Arizona的牛血清白蛋白（BSA）。采用Rinehart和Abui-Haj [9]的方法制备胶体铁（CI）。神经氨酸酶灌注液的制备。在0.1M氯化醋酸钠缓冲液（pH 5.4）的载体中制备浓度为0.5U/ml（40 μ g/ml）和1.0U/ml（80 μ g/ml）的神经氨酸酶灌注液，分别含有0.5 μ g/ml和1.0 μ g/ml SBTI。包括SBTI以消除任何污染的胰蛋白酶活性。灌注液在临使用前和收到酶后几天内制备，因为酶即使储存很短时间也会导致其酶活性显著和可变的降低。
Methods Male Sprague-Dawley rats weighing 150 to 200 g were used (Charles River Breeding Laboratories, North Wilmington, Mas-sachusetts). Materials were purchased as follows: chromatographically purified neuraminidase (Clostridium perfringens) and soybean trypsin inhibitor (SBTI) from Worthington Bio-chemical Corporation, Freehold, New Jersey; puromycin aminonucleoside (PAN)(6-dimethylamino-9 [3'amino-3'deoxyribosyl purine]) from Sigma Chemical Company, St. Louis, Missou-ri; horse spleen ferritin, two times crystallized (cadmium free) from Calbiochem, San Diego, California; and bovine serum albumin (BSA) from Reheis Chemical Company, Phoenix, Arizona. Colloidal iron (CI) was prepared by the method of Rinehart and Abui-Haj [9].Preparation of neuraminidase perfusates. Neuraminidase perfusates, at concentrations of 0.5 U/mi (40 pgIml) and 1.0 U/mi (80 g/m1) were prepared in a vehicle of 0.1 M sodium chloride acetate buffer, pH 5.4, containing 0.5 pg/mi and 1.0 g/ml of SBTI, respectively. SBTI was included to abolish any contaminating tryptic activity. The perfusates were prepared immediately prior to use and within a few days after the enzyme was received since storage of the enzyme for even a short period of time led to a marked and variable reduction in its enzymatic activity.