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Biomedical subjects

C Kirchner

Publications and source records attributed to C Kirchner.

At least 37 records · Page 2Linked to original sources

Purification of uteroglobin using monospecific antibodies coupled to divinylsulphone-activated agarose.

As a model for the isolation of a labile or trace protein, the purification of uteroglobin (UGL) by immunoaffinity chromatography is described. Antibody was isolated from sheep antiserum by immunoprecipitation, and coupled to divinylsulphone-activated agarose (Mini Leak). For the immunoabsorption stage rabbit uterine mucosal scrapings were defatted and incubated directly with the immunosorbent. After washing and desorption, the UGL preparation contained relatively few high molecular weight impurities and these were removed by gel chromatography. Purification was monitored at each step by two-dimensional SDS polyacrylamide gel electrophoresis and immunoelectrophoresis. Furthermore, affinity-purified UGL was tritiated with N-succinimidyl[2,3-3H]propionate and assayed by fluorography. In order to determine absolute UGL concentrations a competitive ELISA was developed.

Animals↗

Active immunization of rabbits against progesterone: increase in hormone levels, and changes in metabolic clearance rates and in genital tract tissues.

11 alpha-Hemisuccinyl progesterone was coupled to rabbit serum albumin and injected into intact male rabbits, and into intact ovariectomized female rabbits, for a period of 52 weeks. The ovariectomized females and the intact males showed better immune responses than the intact females. Specificity of the antisera was tested against both progesterone and the carrier protein, and against 19 other selected steroids. Antibody titres and serum levels of progesterone in all groups, and testosterones in the males, showed characteristic changes with a first maximum 12-18 weeks after the beginning of immunization, a decrease despite booster injections and a second increase after 35-40 weeks. Hormone levels were 5-12 times higher by the end of immunization than before. High antibody titres were correlated with decreased metabolic clearance rates for progesterone, reflecting the binding of the steroid to the antibody. Increased production rates of progesterone in the immunized females and of testosterone in the immunized males showed that the antibody-bound hormone was not available for feedback control. Absence of primordial follicles, hyperplasia of the Leydig cells, decreased spermatogenesis and involution of the seminal vesicle epithelium were interpreted as direct or indirect effects of the removal of free progesterone.

Animals↗

Uteroglobin as progesterone-binding protein in the preimplantation uterine epithelium of the rabbit: histochemical studies.

[3H] progesterone was injected into the uterine lumen of rabbits toward the end of preimplantation period (162 h post coitum). Light-microscopic autoradiography showed accumulation of label in single cell groups of the uterine epithelium. Fluorographs of thin layer chromatograms of steroid extracts indicated the metabolization of progesterone in the uterine tissue. Incubation of uterine sections with fluorescein isothiocyanate-conjugated progesterone-rabbit serum albumin revealed binding sites for this reagent: 162 h post coitum, staining was also localized in single cell groups of the uterine epithelium. Pretreatment with a monospecific antiserum showed uteroglobin to be the binding protein.

Animals↗

Uteroglobin as a progesterone-binding protein in the preimplantation uterine epithelium of the rabbit: biochemical studies.

Progesterone binding was studied in the uterus of rabbits at two different hormonal stages; either after oestrogen priming or a short time before implantation of the blastocyst (162 h post coitum). Uterine cytosols were incubated with [3H]progesterone, or the labelled hormone was injected into the uterine lumen 1 h before killing the animals. Gel filtration, ion exchange chromatography, sucrose gradient centrifugation, isoelectric focusing and saturation analysis indicate that during the period prior to implantation, the uterine progesterone receptor disappears and progesterone binding is performed by uteroglobin. These findings support the hypothesis that the physiological role of uteroglobin in the reproductive process is connected with its hormone-binding ability.

Animals↗

Glycoproteins in rabbit uterus during implantation. Differential localization visualized using 3H-N-acetyl-glucosamine labelling and FITC-conjugated lectins.

The synthesis of glycoproteins in rabbit uterine epithelium during the late preimplantation period was studied using tritiated N-acetylglucosamine. In vivo labelling was achieved by the intra-uterine implantation of agar gel columns containing the precursor. Autoradiography showed the radioactivity to be predominantly localized in the apical cell surfaces, with single cells exhibiting an accumulation of silver grains in their supranuclear cytoplasm. After gel electrophoresis of uterine flushings, activity was mainly found in the beta-glycoprotein fraction. Fluorescein isothiocyanate (FITC)-conjugated wheat-germ agglutinin reacted with the apical cytoplasm and surfaces of the endometrial cells. However, FITC-conjugated concanavalin A exhibited a different binding pattern, reacting first with the basal cytoplasm, and later with the apical cytoplasm. After concanavalin-A staining, single cells exhibited positive vesicles in their lateral and apical parts. These cells may be released into the uterine lumen until 210 h post column. Neither of the lectins reacted with ciliated cells. Concanavalin A showed an affinity for the beta-glycoprotein fraction of the uterine secretion. The results indicate that, although all endometrial cells contain glycoproteins, only a few of these seem to be involved in the synthesis of secretory products.

Acetylglucosamine↗

Further characterization of the beta-glycoprotein of the rabbit uterus.

beta-Glycoprotein was isolated from preimplantation uterine secretions of the rabbit by gel- and ion-exchange chromatography. Two fractions, called DF1 and DF2, were analyzed by isoelectric focusing (IEF) and sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in combination with Western blotting and immunoelectrophoresis. DF1 displayed 21 bands with isoelectric points of pH 5.2-7.6, and DF2 15 bands of pH 4.2-5.7. SDS-PAGE yielded up to 14 bands with major components at molecular weights of 63,000 and 135,000 respectively. Two-dimensional gel electrophoresis of DF2 in combination with Western blotting revealed five groups of proteins of equal molecular weights but with different isoelectric points, indicating immunological identities. Glycosidase activities in uterine secretions before and after implantation were studied and compared with those of the blastocyst fluids. alpha-L-Fucosidase co-eluted with DF1, and beta-N-acetylglycosaminidase was distributed in DF1 and DF2. Both enzymes were localized on isoelectric focusing gels, and N-acetylglucosaminidase was also demonstrated in an immunoprecipitate of DF1.

Acetylglucosaminidase↗

[Changes in the metabolic parameters in patients with acute myocardial infarct in the course of the rehabilitation phase II].

In a prospective study 32 patients with condition after acute myocardial infarction in the rehabilitation phase II were observed concerning the metabolic parameters, beta-lipoproteins, cholesterol, triglycerides, glucose, uric acid and lactate. At the beginning and at the end of the sports therapy the metabolic parameters were tested in rest as well as after exertion. The investigation led to the following results: 1. Increase of the physical efficiency from 134.0 to 142.0 Watt (non-significant); 2. the size of the biochemical parameters (beta-lipoproteins, cholesterol, uric acid, glucose) remained unchanged in the process of rehabilitation. 3. During the exertions themselves (hour of sports therapy, bicycle ergometer) in each case a non-significant increase of the beta-lipoproteins, the cholesterol and the values of uric acid appeared.

Adult↗

Ultrastructural localization of uteroglobin immunoreactivity in rabbit lung and endometrium, and rat ventral prostate.

Recent biochemical studies have demonstrated amino acid sequence homologies between uteroglobin from rabbit endometrium and prostatic binding protein from rat ventral prostate. We have studied the ultrastructural distribution of uteroglobin-immunoreactive material in rabbit lung and endometrium and rat ventral prostate using an uteroglobin antibody raised in guinea pigs. Secretory granules of bronchiolar Clara cells, endometrial non-ciliated cells and rat prostate secretory cells gave a positive immunoreaction when this antibody was used. The results indicate a close relationship of immunoreactive epitopes of proteins present in those secretory cells. The functional properties of these proteins (glycoproteins, steroid binding, androgen-dependent secretion) suggest a close functional relationship, for instance a surface action such as coating, capping, masking or lubrication.

Androgen-Binding Protein↗

Purification and immunohistology of a glycoprotein secreted from the rabbit uterus before implantation.

Two antigens of the beta-glycoprotein fraction from rabbit uterine secretion of the seventh day post coitum were purified firstly by gel filtration on Sephadex G 150, then either by ion exchange chromatography on DEAE Sephacel or chromatofocusing on PBE 94. By the use of a specific antiserum, raised in female sheep, two antigens with alpha 2- and beta 2-mobility in agar gel electrophoresis could be demonstrated. Immunohistochemical staining of the uterine epithelium at the seventh and eighth day post coitum showed the antigens to be localized in a ciliated cell type of conspicuous shape, which is supposed to be the site of synthesis. Stain accumulated mainly in the apical part of the cell, but there were also small deposits around the nucleus.

Animals↗

Progesterone dependent uptake of uteroglobin by rabbit endometrium.

Unlabeled or 3H-labeled UGL, isolated from rabbit lungs, was injected intraluminally into the uteri of ovariectomized low dose progesterone substituted and unsubstituted animals as well as into mated animals on the fifth day of pregnancy. It was offered to the endometrium, in this last case, in competition with endogenously synthesized UGL. 3H-labeled IgG served as a control. The animals were killed 24 h after the experiment. Immunohistograms and autoradiograms showed that UGL was taken up by the whole endometrium of the ovariectomized animals but only when progesterone was present. In the preimplantation uterus, uptake was limited to clusters of smaller cells accumulated in the lumen adjacent to the epithelium, whereas IgG remained within the uterine lumen, forming a dense layer on the endometrial surface.

Animals↗

Influence of vitamin A deficiency on the excretion of uromucoid and other substances in the urine of rats.

Male Fü-albino rats were weaned at the age of four weeks and maintained on a vitamin A-deficient diet. When they were 14-18 and 21-26 weeks old, the concentration of uromucoid, calcium and other substances possibly important for the pathogenesis of urinary calculi were determined. Reduced uromucoid excretion with hypercalciuria and reduced phosphate levels were observed. Subsequent examination of the kidneys did not demonstrate the presence of nephrocalcinosis or lithiasis. The relation between vitamin A, the synthesis of uromucoid and AMPS and calcium metabolism in the renal tubules is discussed.

Albuminuria↗

Immunohistochemical localization of secretory proteins in the endometrial epithelium of the rabbit.

Proteins of uterine fluid and lung homogenates of the rabbit were separated by gel and ion exchange chromatography. Purified protein fractions were used for immunisation and antiserum production. By means of several absorptions, six monospecific antisera against uteroglobin and five other proteins were obtained. Using immunohistochemistry, four of them could be localised in the uterine epithelium from oestrus and the first and the seventh day post coitum, and also in the blastocyst. The present study indicates the involvement of different endometrial cells in the synthesis and release of the various proteins of uterine secretion.

Animals↗

Uteroglobin-like antigen in the pulmonary epithelium and secretion of the lung.

The immunological identity of a uteroglobin-like protein, occurring in respiratory tract secretions and tissue, with uteroglobin from rabbit endometrial secretion is demonstrated. A uteroglobin-like antigen has been localized in bronchial epithelial cells and in bronchioles by immunofluorescence. This secretory protein is, in contrast to the authentic uteroglobin, hormone-independent, as far as estrogens and progesterone are concerned. The possible significance of comparative studies on uteroglobin and the uteroglobin-like antigen is discussed, taking into account cytological, endocrinological, and molecularbiological aspects.

Animals↗

Influence of seminal plasma proteins on motility of rabbit spermatozoa.

Washed rabbit spermatozoa of freshly collected ejaculates were incubated in different fractions of seminal plasma proteins and examined for motility. Incubation media were protein fractions obtained by gel and ion-exchange chromatography. Motility was best maintained in the fractions which contained albumin, and rabbit serum albumin was more effective than bovine or human serum albumin. Concentrations of less than 4 mg rabbit serum albumin/ml resulted in markedly decreased motility.

Animals↗

A student-run course in interprofessional relations.

The topic of working relationships among health professionals is one which has long been neglected in professional school curricula. During the winter of 1976 students from four health professions--medicine, nursing, occupational therapy, and physical therapy--initiated a course designed to improve their knowledge of one another's training and skills, with the larger aim of preparing themselves to work in health care teams. The nine weekly meetings are described in terms of their content and students' reactions. In the discussion the underlying themes of legal responsibilities and of sex-roles as factors in health care team functioning are identified. Suggestions are made about format and techniques that seem to be useful for courses of this type.

Education, Medical, Undergraduate↗