Determination of endogenous serum proteins in normal and oedematous brain tissue of cat by rocket and crossed immunoelectrophoresis

Summary A quantitative method for determination of endogenous serum proteins has been established and tested in an experimental model of peritumoural brain oedema in cats. Rocket and crossed immunoelectrophoresis were applied for determination of total serum proteins, albumin, IgG and haemoglobin in blood and brain homogenates. Modifications such as the use of non-ionic detergents and of antisera with different specificity were established for each antigen under investigation. The content of total serum proteins, albumin and IgG was substantially higher in tumour and peritumoural brain tissue than in the non-oedematous brain. The measurement of haemoglobin allowed the calculation of blood volume and, in consequence, the differentiation between intra- and extravascular serum proteins. The results are in line with earlier measurements obtained by different analytical methods and demonstrate that the present technique provides a reliable approach for the quantitative assessment of serum protein extravasation.     

Pancuronium binding by serum proteins

Pancuronium has not been suspected of binding to serum proteins. Experimental evidence is presented of strong binding to gamma globulin, and of moderate binding to albumin. No evidence could be found of binding to alpha of beta globulin fractions. Less than 13% of a dose in the clinical range would be unbound and active.     

A serum biochemical profile of normal pregnancy

Serum sodium, potassium, chloride, magnesium, calcium, phosphate, urea, urate, creatinine, carbon dioxide, total alkaline phosphatase, total protein and albumin levels during pregnancy were studied in groups of subjects at less than 14 weeks' gestation, at 14-20 weeks, at 20-28 weeks and at 36-40 weeks. These were compared with levels in a nonpregnant control group matched for age and sex. Serum potassium, sodium and chloride levels remained within the normal range, while calcium, phosphate, total protein and albumin levels decreased progressively after the first trimester. Magnesium levels decreased progressively from early pregnancy to term, and urea, creatinine and carbon dioxide levels all decreased markedly throughout pregnancy. Urate levels were markedly decreased in early pregnancy but returned to normal at term, and the alkaline phosphatase values demonstrated the expected marked increase to term. The fact that all these values were not equally affected suggests that haemodilution cannot be held responsible for all serum changes during pregnancy. Individual results are discussed and a biochemical profile is suggested.     

Abnormalities of serum proteins following thermal injury

Suppressive factors were found to be present in serum and considered to be one of the causes of the suppression of body defences following thermal injury. The purpose of this study was to find out any abnormality of the postburn serum proteins by the comparison of burned and normal sera using immunoprecipitation in gel. The results of SDS-polyacrylamide gel electrophoresis, crossed immunoelectrophoresis, tandem crossed immunoelectrophoresis, and fused rocket immunoelectrophoresis showed that differing precipitation bands or peaks existed between burned and normal sera, and abnormal constituents with relatively low molecular weight were present in the burned serum.     

血清CEA阴性结直肠癌患者血清差异蛋白质的初步筛选

目的分析血清CEA阴性结直肠癌患者、血清CEA阳性结直肠癌患者与正常人血清中差异表达蛋白,筛选潜在的与血清CEA阴性结直肠癌患者诊断及预后密切相关的血清蛋白质。方法血清CEA阴性结直肠癌患者、血清CEA阳性结直肠癌患者与正常人空腹血清标本各11例分别等量混合成3组样本,利用双向凝胶电泳（2-DE）及基质辅助激光解吸电离飞行时间质谱（MALDI_TOF-MS／MS）技术筛选及鉴定差异表达的蛋白点,并对其部分蛋白进行生物学分析。结果筛选出3组间表达量差异2倍以上差异蛋白点共13个,进行MALDI-TOF-MS／MS分析,鉴定出：①血清CEA阴性结直肠癌患者与正常人血清比较,表达下调的蛋白有纤维胶凝蛋白2前体,纤维胶凝蛋白3,载脂蛋白L1,间a胰蛋白酶抑制剂轻链H4,转甲状腺素蛋白,表达上调的蛋白为免疫球蛋白lambda轻链。②血清CEA阳性结直肠癌患者与正常人血清比较,表达下调的蛋白有纤维胶凝蛋白2前体,纤维胶凝蛋白3,载脂蛋白L1,间a胰蛋白酶抑制剂轻链H4,转甲状腺素蛋白,表达上调的蛋白为：载脂蛋白E,结合珠蛋白。③血清CEA阴性结直肠癌患者与血清CEA阳性结直肠癌患者的1个差异点,经鉴定为免疫球蛋白lambda轻链。结论血清CEA阴性结直肠癌患者与血清CEA阳性结直肠癌患者、正常人血清的蛋白质表达谱表现出一定差异,这些差异蛋白有可能成为血清CEA阴性结直肠癌患者诊断与判定预后的血清标志物。     

正常人血清前列腺特异性抗原水平的研究

目的：评价年龄与血清前列腺特异性抗原（PSA）水平的关系。方法：采用免疫放射分析（IRMA）测定226例正常男性各年龄组血清PSA水平。结果：30～39岁组血清PSA为（0．88±O．47）ng／ml，40～49岁组为（0．87±0．34）ng／ml，50～59岁组为（0．99±0．57）ng／ml，60～69岁组为（1．12±0．78）ng／ml，＞70岁组为（1．12±0．83）ng／ml，各组间血清PSA浓度无显著性差异（P＞0．05）。结论：当前列腺体积在正常范围内时，血清PSA水平与年龄无直接关系。     

Alterations in serum proteins following thermal injuries

Serum suppressive factors were proved to be present and considered to be one of the causes to bring about the suppression of body defence following thermal injury. As a preliminary step to ascertain the presence of immunosuppressive substances, we tried to find out any abnormality in the postburn serum proteins by comparison of burned and normal sera by means of immuno-precipitation-in-gel. Results of SDS-polyacrylamide gel electrophoresis, crossed immunoelectrophoresis, tandem crossed immunoelectrophoresis, and fused rocket immunoelectrophoresis showed that differences of precipitation bands or peaks existed between burned and normal sera, and abnormal constituents with relatively low molecular weight were present in the burned serum.     

Noncollagenous proteins in normal and pathological human bone

The concentrations of alpha 2HS-glycoprotein, albumin, and sialic acid were measured in the bone of 28 normal individuals and 6 patients with osteogenesis imperfecta, 3 patients with Paget's disease, and 4 patients with either renal osteodystrophy, osteoporosis, osteomalacia, or osteopetrosis. The concentration of alpha 2HS-glycoprotein in neonatal bone was 3 X higher than in bone from children and 7 X higher than in adult bone. The concentrations of albumin and sialic acid in neonatal bone were 1.5 X higher that in bone from children and twice the levels in adult bone. The concentrations of alpha 2HS-glycoprotein, albumin and sialic acid were above the normal mean values in the patients with osteogenesis imperfecta, and 4 patients had significantly raised levels of one or more of these proteins. The concentrations of these noncollagenous proteins were also significantly raised in Paget's disease, and the alpha 2HS-glycoprotein was significantly raised in renal osteodystrophy. The lowest level of alpha 2HS-glycoprotein was in osteopetrotic bone and the lowest levels of albumin and sialic acid were in osteoporotic bone. The results of this study suggest that the concentrations of alpha 2HS-glycoprotein, albumin, and sialic acid in bone are related to the rate of bone turnover.     

Noncollagenous proteins in normal and pathological human bone

Summary The concentrations of α₂HS-glycoprotein, albumin, and sialic acid were measured in the bone of 28 normal individuals and 6 patients with osteogenesis imperfecta, 3 patients with Paget's disease, and 4 patients with either renal osteodystrophy, osteoporosis, osteomalacia, or osteopetrosis. The concentration of α₂HS-glycoprotein in neonatal bone was 3 × higher than in bone from children and 7 × higher than in adult bone. The concentrations of albumin and sialic acid in neonatal bone were 1.5 × higher that in bone from children and twice the levels in adult bone. The concentrations of α₂HS-glycoprotein, albumin and sialic acid were above the normal mean values in the patients with osteogenesis imperfecta, and 4 patients had significantly raised levels of one or more of these proteins. The concentrations of these noncollagenous proteins were also significantly raised in Paget's disease, and the α₂HS-glycoprotein was significantly raised in renal osteodystrophy. The lowest level of α₂HS-glycoprotein was in osteopetrotic bone and the lowest levels of albumin and sialic acid were in osteoporotic bone. The results of this study suggest that the concentrations of α₂HS-glycoprotein, albumin, and sialic acid in bone are related to the rate of bone turnover.     

Nephrotoxicity of vancomycin in patients with normal serum creatinine

Sir, The reported rate of nephrotoxicity of vancomycin (VCM) hasbeen 7–16%. It can reach 35% with concurrent aminoglycosidesand is associated with serum concentration >40 µg/ml[     

Trauma serum suppresses superoxide production by normal neutrophils

The effect of trauma serum on superoxide production by normal neutrophils was studied in 47 serum samples from 18 patients with multiple trauma. Ten patients became septic and eight patients remained nonseptic. Incubation in trauma serum significantly suppressed superoxide production by normal neutrophils compared with incubation in normal serum: 3.6 +/- 1.44 vs 4.04 +/- 1.64 nmole of superoxide produced by 10(6) neutrophils (mean +/- SD). There was no difference in the suppressive effect between septic and nonseptic trauma serum samples. The chemiluminescence response of normal neutrophils was likewise suppressed following incubation in trauma serum compared with incubation in normal serum. The chemiluminescence response correlated with superoxide reduction of cytochrome C. In addition, the chemiluminescence response was significantly less in septic-trauma serum than in nonseptic-trauma serum. Suppressive serum was found to inhibit the neutrophil-membrane depolarization response to latex particles, as measured by flow cytometry. We conclude that trauma serum suppresses superoxide production by normal neutrophils, and that such suppression can be detected reliably using the clinically applicable technique of chemiluminescence. A normal chemiluminescence response excludes serum-mediated suppression of neutrophil superoxide production. In addition, chemiluminescence may be of value in detecting altered resistance to sepsis following injury, while superoxide determinations do not seem to be helpful in this regard. The mechanism of action of the suppressor may involve reversible inhibition of membrane depolarization necessary for the production of bactericidal oxygen species.     

血清蛋白质组学研究中高丰度蛋白的去除

目的 建立血清蛋白质组学研究中高丰度蛋白的去除方法并观察其去除效果.方法 应用Agilent公司去除人14种高丰度蛋白的多重亲和排除系统(Human 14 Multiple Affinity RemovalSystem),去除血清中自蛋白、IgG和抗胰蛋白酶等14种高丰度蛋白,并用SDS-PAGE评价去除的效果.结果 建立血清中高丰度蛋白的去除方法,可去除白蛋白等14种高丰度蛋白.SDS-PAGE图谱中去除高丰度蛋白后条带更清晰,部分低丰度蛋白得以显现.结论 成功建立血清中高丰度蛋白的去除方法,可为下游的蛋白质组学技术奠定基础.

Abstract：

Objective To establish the method for removing high-abundance proteins in serum proteomics research and evaluate the removal efficiency of the high-abundance proteins. Methods Human 14 Multiple Affinity Removal System of Agilent Technologies was applied to eliminate the high-abundance proteins in serum, such as albumin, IgG and α-l-antitrypsin, etc. The efficiency of removal was evaluated by 1-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Results The method for removing serum high-abundance proteins was established. The protein lanes of SDS-PAGE map were clearer and some low-abundance proteins were visualized after depletion. Conclusion An effective method of removing high-abundance proteins in serum is established, which provids basis for further strategies in proteomics research.