The impact of different fixation procedures on staining of macromolecules in a microtiter system

Summary The effects of formaldehyde, glutaraldehyde, methanol, Clarke's fixative and microwave irradiation on the quantitative staining of proteins (Naphthol Yellow S) and nucleic acids (Ethyl Green—Pyronin) in a cell culture system have been investigated. Overall, glutaraldehyde rapidly yielded the highest and most consistent levels of staining when compared to all other chemical fixatives. Although microwave irradiation was found to be uneven, 4 min exposure to 700W was found to give higher levels of protein staining than those achieveable with glutaraldehyde. Time-dependent processes were observed with all procedures. In addition, dissociations in the trends of protein and nucleic acid staining were observed. It is suggested that these results domonstrate fixation events that have not previously been resolved from the effects of reagent penetration into tissue blocks.     

Transport of an Mr approximately 300,000 Plasmodium falciparum protein (Pf EMP 2) from the intraerythrocytic asexual parasite to the cytoplasmic face of the host cell membrane

The profound changes in the morphology, antigenicity, and functional properties of the host erythrocyte membrane induced by intraerythrocytic parasites of the human malaria Plasmodium falciparum are poorly understood at the molecular level. We have used mouse mAbs to identify a very large malarial protein (Mr approximately 300,000) that is exported from the parasite and deposited on the cytoplasmic face of the erythrocyte membrane. This protein is denoted P. falciparum erythrocyte membrane protein 2 (Pf EMP 2). The mAbs did not react with the surface of intact infected erythrocytes, nor was Pf EMP 2 accessible to exogenous proteases or lactoperoxidase-catalyzed radioiodination of intact cells. The mAbs also had no effect on in vitro cytoadherence of infected cells to the C32 amelanotic melanoma cell line. These properties distinguish Pf EMP 2 from Pf EMP 1, the cell surface malarial protein of similar size that is associated with the cytoadherent property of P. falciparum-infected erythrocytes. The mAbs did not react with Pf EMP 1. In one strain of parasite there was a significant difference in relative mobility of the 125I-surface-labeled Pf EMP 1 and the biosynthetically labeled Pf EMP 2, further distinguishing these proteins. By cryo-thin-section immunoelectron microscopy we identified organelles involved in the transit of Pf EMP through the erythrocyte cytoplasm to the internal face of the erythrocyte membrane where the protein is associated with electron-dense material under knobs. These results show that the intraerythrocytic malaria parasite has evolved a novel system for transporting malarial proteins beyond its own plasma membrane, through a vacuolar membrane and the host erythrocyte cytoplasm to the erythrocyte membrane, where they become membrane bound and presumably alter the properties of this membrane to the parasite's advantage.     

猫颈动脉区压力和化学感受性刺激对巨细胞旁外侧核单位放电的影响

在31只氯醛糖和氨基甲酸乙酯麻醉的猫,观察了选择性激活颈动脉压力和化学感受器对巨细胞旁外侧核(PGL)单位放电的影响。136个PGL自发放电单位中,有84个在激活颈动脉压力感受器(BA)(新福林,1—2μg/kg,iv)和/或激活颈动脉化学感受器(CA)(nicotine 5—20μg,溶于0.25—0.5ml生理盐水中,注入甲状腺动脉)时,放电频率有变化。在这些有反应的单位中,16个仅对CA起反应(11个兴奋、5个抑制);54个以各种组合方式对CA和BA都起反应,其中以CA引起兴奋反应而BA引起抑制反应的占比例最大;14个仅对BA起反应(7个兴奋,7个抑制)。在定位分布上,那些只对CA起反应的单位多位于PGL的腹侧部份;仅对BA起反应的单位则位于对CA起反应单位的较背侧;对BA和CA均起反应的单位介于上述两者之间或在较深区域。这些结果表明,颈动脉区压力和化学感受器活动传入到PGL,并会聚在其中一些神经元上。 在PGL内全部有反应的单位中,68个对激活颈动脉压力感受器起反应,其中兴奋的29个,抑制的39个(P>0.05);70个对激活颈动脉化学感受器起反应,其中48个兴奋,22个抑制(P<0.005)。这些结果提示,BA对PGL神经元引起兴奋和抑制两种效应,而CA则诱发兴奋为主的反应。     

A new device to measure the structural properties of the femur-anterior cruciate ligament-tibia complex

Previous studies of biomechanical properties of femur-anterior cruciate ligament-tibia complex (FATC) utilized a wide variety of testing methodologies, particularly with respect to ligament orientation relative to loading direction. A new device was designed and built to test the anterior-posterior displacement of the intact porcine knee at 30 and 90 deg of flexion, as well as the tensile properties of the FATC at any loading direction and flexion angle. Tensile tests were performed with the knees at 30 and 90 deg of flexion with the loading direction along either the axis of the tibia (tibial axis) or the axis of the anterior cruciate ligament (ligament axis). The results showed that the stiffness, ultimate load and energy absorbed were all significantly increased when the FATC was tested along the ligament axis. This study demonstrates the importance of alignment in the evaluation of the biomechanical characteristics of the femur-ACL-tibia complex.     

心房钠尿因子对麻醉家兔局部血流的影响

在42只麻醉家兔,观察了静脉注射心房肽Ⅱ(AtriopeptinⅡ,APⅡ)对局部血流量以及动脉内注射 AP Ⅱ 对局部血管阻力的影响。结果如下:(1)静脉注射 APⅡ(30μg/kg)5min后,平均动脉压(MAP)降低11.0±1.5mmHg(n=8,M±SE,下同),与溶剂对照组相比有明显差异(P相似文献