Purification and chemical modification of porcine bone morphogenetic protein

Implantation of porcine bone morphogenetic protein (pBMP) in the muscle induces differentiation of mesenchymal-type cells and results in endochondral bone formation. pBMP was isolated from porcine demineralized bone matrix and purified by hydroxyapatite chromatography, Sephadex G75 gel filtration, preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), preparative isoelectric focusing (IEF), and chromatofocusing fast protein liquid chromatography (FPLC). Porcine BMP has an MW of 26 K and a range of pI from 4.65 to 4.73 determined by SDS-PAGE and IEF, respectively. Reconstitution with the citrate buffer supernatant fraction enables as little as 50 micrograms of the soluble pBMP fractions to induce osteogenesis in an in vivo assay. Chemical modification studies indicate that the osteoinductive potential of the pBMP molecule depends on tyrosine, carboxyl groups, and disulfide bonds and can be increased by modification of sulfhydryl groups. Modification of arginine and tryptophan has no effect on bioactivity. By pepsin-limited proteolysis, fragments of pBMP with an MW of 6-14 K show definite, although reduced, BMP activity.     

Inhaled natural human interferon alpha induces bronchospastic reactions in asthmatics.

There is a relationship between viral infection and aggravation of bronchial asthma symptoms. Viruses belong to the strongest and fast acting inducers of interferon (IFN), which, in turn, may enhance histamine release from effector cells. We have administered IFN intrabronchially to 49 patients suffering from bronchial asthma. The impairment of lung ventilation with significant decrease of forced expiratory volume in one second (FEV1) have been observed in majority of the patients who inhaled crude or purified IFN preparations.     

战士体内VitB_1、VitB_2储备水平检测

选择南方热区某特种部队战士５２人，分两组分别口服５ｍｇＶｉｔＢ１和ＶｉｔＢ２，收集４ｈ尿液，用荧光法进行负荷试验。结果：ＶｉｔＢ１组，所测２９人中，１５人缺乏，９人不足，２人正常，３人充裕；ＶｉｔＢ２组，所测２３人中，１１人缺乏，１２人不足，反映该部队全年约５％的发病是由于ＶｉｔＢ１、ＶｉｔＢ２缺乏不足所致。其原因为膳食摄入不够     

白细胞介素—2新的功能位点及其中枢镇痛作用

白细胞介素－２（ＩＬ－２）不仅是重要的免疫调节因子，而且还具有重要的中枢调节作用。本实验以钾离子透入引起大鼠甩尾反应为指标，发现侧脑室注射ＩＬ—２能显著提高动物痛阈，并能被纳洛酮所阻断，表示ＩＬ－２的中枢镇痛作用可能与阿片受体有关。利用基因定位突变技术获得的无免疫活性ＩＬ－２实查体仍具有中枢镇痛作用，表明ＩＬ—２分子上发挥镇痛和免疫调节作用的功能位点是相互独立的。纳洛酮能够阻断ＩＬ—２的中枢镇痛作用，而不能影响ＩＬ—２增殖ＣＴＬＬ－２细胞的作用，提示ＩＬ－２发挥镇痛和免疫调节作用可能通过不同的受体途径。ＩＬ－２分子中第４５位Ｔｙｒ残基突变为Ｖａｌ后，虽仍保留了免疫活性，但丧失了镇痛功能，表示４５位Ｔｙｒ残基是ＩＬ—２发挥中枢镇痛功能的关键残基之一。我们推测ＩＬ—２的镇痛功能位点可能在ＩＬ—２分子中第４５位Ｔｙｒ残基附近区域。     

纤维组织粘合剂的实验研究

本文应用“冷沉淀”法制备纤维组织粘合剂。对纤维组织粘合剂中主要成份和含量进行了测定,并测定了主要理化性质,建立了动物实验模型,观察了实验兔对纤维粘合剂的反应。     

Immunochemical and biochemical characterization of purified canine interferon-gamma. Production of a monoclonal antibody, affinity purification, and its effect on mixed lymphocyte culture and mixed lymphocyte kidney culture reactions.

We have advanced the hypothesis that the primary autolymphoproliferative response of dog T cells in mixed lymphocyte kidney cultures (MLKC) results from their recognition of tissue-specific (kidney-associated) antigen(s) presented in conjunction with class II MHC antigens. Lymphocyte culture-derived supernatants had been found previously to upregulate class II antigen expression on kidney cells and enhance T cell activation. In the present study we have isolated and characterized dog IFN-gamma, a class II-inducing substance that is secreted in the culture supernatant of activated T lymphocytes. Dog IFN-gamma was induced with A-23187 and PMA and purified stepwise using controlled-pore glass, Mono Q anion exchange chromatography, and Superose 6-gel filtration on FPLC. The purification resulted in two molecules of 42 Kd and 31 Kd molecular weights. An IgG1 monoclonal antibody was engendered to these molecules. With this mAb reagent, in immunochemical experiments, we have developed a sensitive ELISA and a method for purifying dog IFN-gamma by affinity chromatography. Species specificity studies indicated that purified dog IFN-gamma reacted with a polyclonal rabbit antihuman IFN-gamma, but not with a mAb to human IFN-gamma. However, the antidog IFN-gamma mAb that was generated also reacted with recombinant human IFN-gamma. In in vitro biological studies, the purified IFN-gamma (two mol. wt. species) upregulated the expression of canine class II MHC molecules on dog tubular epithelial cells and the dog kidney epithelial cell line (MDCK). The antidog IFN-gamma mAb blocked T cell proliferative response to kidney cell and, by inference, the interaction between endogenously released IFN-gamma in vitro with its cell surface receptor, thus inhibiting the induced upregulation of class II. Interestingly, although antidog IFN-gamma markedly blocked the MLKC (10 micrograms mAb/well), there was no effect on the allogeneic MLC. This observation indicates that the cytokine IFN-gamma may be a uniquely key substance amplifying the immune response of T cells to tissue-associated antigens on surrogate antigen-presenting cells that require induced upregulation of class II MHC antigen expression (MLKC), in contrast to reactions in which these antigens are already constitutively expressed on the antigen-presenting cells (mixed lymphocyte culture).     

Application of a 3D volume 19FMR imaging protocol for mapping oxygen tension (pO2) in perfluorocarbons at low field

A limited flip angle gradient-echo 3D volume acquisition imaging protocol for mapping partial pressure of oxygen (pO₂) in perfluorocarbon compounds (PFCs) at low field (0.14 T) is presented. The pO₂ measurement method is based on the paramagnetic effect of dissolved molecular oxygen (O₂) which reduces the PFC ¹⁹F T_1? Specific objectives related to imaging of PFCs through use of the protocol include improved image signal-to-noise characteristics and elimination of ¹⁹F chemical shift artifacts. A parametric Wiener deconvolution filtering algorithm is used for suppression of ¹⁹F chemical shift artifacts. Application of the protocol is illustrated in a series of calculated pO₂ maps of a gas equilibrated, multi-chamber phantom containing perfluorotributylamine (FC-43). The utility of the protocol is demonstrated in vivo through images of a commercially available perfluorocarbon based blood substitute emulsion containing FC-43 sequestered in the liver and spleen of a rat.     

Assessment of carbohydrate tolerance in pregnancy

A review is given of the various methods of assessing carbohydrate tolerance in pregnancy. Oral glucose tolerance screening and diagnostic tests have been in use for more than 25 years. They are easily administered, relatively inexpensive, and present reasonable sensitivity; therefore, they continue to be used quite extensively. However, lack of reproducibility of the results and side effects such as nausea, vomiting, and headache have led to the use of alternate methods including glucose polymer (Polycose) and standard breakfast meals. These methods have been reported to present satisfactory results in clinical practice. Glycosylated hemoglobin (HbA1c) and fructosamine assays are also alternate forms of testing carbohydrate metabolism HbA1c measurement have been proven insensitive as a screening test for gestational diabetes, while their use as an index of overall glucose control remains valuable. The role of fructosamine in the assessment of carbohydrate intolerance remains controversial with conflicting claims made by various investigators regarding its sensitivity in detecting gestational diabetes and its response to alterations in glycemic control. In this review, the relative advantages and disadvantages of each glucose tolerance test are discussed and recommendations are given regarding their utility in pregnancy.     

Teriparatide increases bone formation in modeling and remodeling osteons and enhances IGF-II immunoreactivity in postmenopausal women with osteoporosis.

Transiliac bone biopsies were obtained from 55 women treated with teriparatide or placebo for 12-24 months. We report direct evidence that modeling bone formation at quiescent surfaces was present only in teriparatide-treated patients and bone formation at remodeling sites was higher with teriparatide than placebo. INTRODUCTION: Recombinant teriparatide [human PTH(1-34)], a bone formation agent for the treatment of osteoporosis when given once daily subcutaneously, increases biochemical markers of bone turnover and activation frequency in histomorphometry studies. MATERIALS AND METHODS: We studied the mechanisms underlying this bone-forming action of teriparatide at the basic multicellular unit by the appearance of cement lines, a method used to directly classify surfaces as modeling or remodeling osteons, and by the immunolocalization of IGF-I and IGF-II. Transiliac bone biopsies were obtained from 55 postmenopausal women treated with teriparatide 20 or 40 microg or placebo for 12-24 months (median, 19.8 months) in the Fracture Prevention Trial. RESULTS: A dose-dependent relationship was observed in modeling and mixed remodeling/modeling trabecular hemiosteons. Trabecular and endosteal hemiosteon mean wall thicknesses were significantly higher in both teriparatide groups than in placebo. There was a dose-dependent relationship in IGF-II immunoreactive staining at all bone envelopes studied. The greater local IGF-II presence after treatment with teriparatide may play a key role in stimulating bone formation. CONCLUSIONS: Direct evidence is presented that 12-24 months of teriparatide treatment induced modeling bone formation at quiescent surfaces and resulted in greater bone formation at remodeling sites, relative to placebo.     

Innervation of the trapezius muscle: is cervical contribution important to its function?

OBJECTIVE: To study the effect of the cervical plexus and the accessory nerve to the function of the trapezius muscle. STUDY DESIGN AND SETTING: Eighteen adult male Sprague-Dawley rats were randomly divided into three groups. The neurotomies were performed in the left sides and the right sides served as within-subject controls. In group A, the accessory nerve was transected. The C2-5 were transected in group B, and both of the accessory nerve and C2-5 were cut in group C. The electrophysiologic, myophysiologic, and histologic changes of the muscles were measured. RESULTS: There were significant differences (P < 0.05) between the three groups in the recovery rates of the transverse area of the muscles. The CMAP recorded from the experimental sides in group B were similar to the control sides. The values of the maximum tension of the tetanus contraction between the two sides showed no differences either (P > 0.05). CONCLUSION: The accessory nerve supplies the most important motor input to the trapezius. Motor innervations of the cervical plexus are not very significant.