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Biomedical subjects

H Haupt

Publications and source records attributed to H Haupt.

At least 109 records · Page 6Linked to original sources

Properties of acute phase proteins of human plasma.

Almost 50 years ago the term acute phase serum was used for the first time by Avery and his colleagues to characterize serum from patients in an acute state with inflammatory diseases and which contained C-reactive protein. In the meantime a variety of proteins was detected in human blood plasma which are subject to the acute phase response. About a fifth of the until today more than hundred isolated and well characterized plasma proteins are to be classified as acute phase proteins. In human plasma they exist in many various quantities. There is no functional relationship among acute phase proteins. They partly belong to the coagulation system or to the complement system or they are transport proteins and inhibitors. There's also no relationship regarding their physico-chemical properties. The majority of the acute phase proteins are glycoproteins of different molecular weight. However, CRP and SAA, which serum concentration can rise to one thousandfold above normal during an acute inflammatory process, are pure polypeptides. The interpretation of the present knowledge about the functional activities of the acute phase proteins has led to the opinion that they probably all play a role during inflammatory processes. It is believed that the acute phase response represents a physiological mechanism which provides the site of inflammation via the tissue fluid with increased concentrations of these proteins. The majority of the acute phase proteins is synthesized in hepatocytes. The stimulation of the synthesis is probably mediated through cytokines (IL1) which are released from macrophages at the site of tissue damage.

Acute-Phase Proteins↗

Biochemical differences between perilymph, cerebrospinal fluid and blood plasma in the guinea pig.

Seven selected biochemical substances were systematically analyzed in perilymph (PL), cerebrospinal fluid (CSF) and blood plasma or serum of anesthetized guinea pigs in order to compare the composition of these fluids with regard to the origin and biochemical nature of PL. Compositional differences between the fluids were expressed by PL/CSF and PL/plasma concentration ratios of the chemical components measured. Contrary to the prevailing notion, the results suggest that the specific biochemical composition of PL corresponds neither to that of CSF nor to that of a simple ultrafiltrate of blood plasma. Furthermore, except for the total protein concentration, no significant differences were found in the composition of PL between the scala tympani and scala vestibuli.

Animals↗

[Effect of air pollution on respiratory tract diseases in small children. Morbidity rate in residential areas with varying degrees of severe pollution].

In a retrospective study of hospitalised patients (children aged between 0 and 4 years) covering four years the dependence of the incidence of croup (infectious laryngitis) and obstructive bronchitis on air pollution was investigated. For this reason the incidence of morbidity in residential areas with various levels of SO2 and sedimenting dust were compared. The results show a definitive connection between a high level of SO2 and dust, respectively and a high rate of morbidity. This connection was particularly marked in areas with simultaneous high levels of both SO2 and dust. The effects of confounding factors, such as infection incidence, meteorological influences, distances from hospital, social status, especially the percentage of foreign guest worker families and population density were investigated and separated from air pollution effects.

Air Pollution↗

The structure of the carbohydrate units of human plasma galactoglycoprotein determined by 500-megahertz 1H NMR spectroscopy.

Human plasma galactoglycoprotein (Mr = 81,000) which was recently identified and characterized (Schmid, K., Mao, S. K. Y., Kimura, A., Hayashi, S., and Binette , J. P. (1980) J. Biol. Chem. 255, 3221-3226) was found to possess an unusually high carbohydrate content (76%) comprised of NeuAc, Gal, Man, Fuc, GalNAc, and GlcNAc. The aim of the present investigation was to elucidate the primary structure of the oligosaccharide units of this protein. For the study of the O-glycosidic oligosaccharide chains, the protein was subjected to beta-elimination and the resulting oligosaccharide preparations were analyzed by 500-MHz 1H NMR spectroscopy. The structure of the predominant glycan, a hexasaccharide: (formula; see text) and that of a tetrasaccharide: NeuAc alpha(2----3)Gal beta(1----3)[NeuAc alpha (2----6)]GalNAc-ol, were determined. The protein possesses approximately 40 hexasaccharides and 3 tetrasaccharides/mol. For the isolation of its N-glycosidic oligosaccharide chains, the protein was exhaustively digested with proteases followed by chromatography of the desialyzed resulting glycopeptide fraction on concanavalin A-Sepharose. 500-MHz 1H NMR spectroscopy of the obtained preparations revealed the presence of 3 diantennary N-glycosidic chains that are extended with a Fuc residue at the Asn-linked GlcNAc.

Carbohydrate Sequence↗

On sources of error in the biochemical study of perilymph (guinea-pig).

Contamination of perilymph with other fluids (cerebrospinal fluid, tissue fluid, blood, endolymph) together with sampling, anaesthesia, surgical intervention or food intake of the animals may considerably affect the analytical result. The numerous possible artefacts seem to be the main reason why varying values are given in the literature for the same chemical component of perilymph. This is also partly true of cerebrospinal fluid and blood. The effect of some sources of error on selected chemical components of perilymph, cerebrospinal fluid and blood is briefly summarized.

Anesthesia↗

[Total lactate dehydrogenase activity of perilymph, plasma and cerebrospinal fluid in unstressed and noise stressed guinea pigs].

The total activity of lactate dehydrogenase (LDH) of perilymph (PL), plasma, and cerebrospinal fluid (CSF) of unexposed and sound-exposed guinea pigs was examined with due consideration of the principal sources of error. To test the purity of PL samples, protein, potassium, and sodium were determined simultaneously (Table 1). The LDH was analysed fluorometrically. It was found that there are considerable differences in the LDH activities of PL, CSF, and plasma (Table 2). The mean activity of PL was three to four times higher than that of CSF and only about half that of plasma. No significant difference was found between the PL in scala tympani and scala vestibuli. The most frequent LDH values of the individual fluids (Fig. 2) were somewhat lower than the mean values calculated. Immediately (less than or equal to 60 min) after exposing the animals to wide-band noise at an intensity of 140 dB SPL for 10 min, the mean PL values of the scala tympani and scala vestibuli were found to be somewhat higher than in unexposed animals (Table 3). Eighteen hours after the exposure, slightly higher activity was only detectable in PL of the scala vestibuli. The differences were not found to be significant. The LDH values of CSF and plasma remained unchanged both less than or equal to 60 min and 18 h after noise exposure.

Animals↗

[Postmortem changes in the perilymphatic lactate and pyruvate concentrations of guinea pigs. (author's transl)].

Lactate and pyruvate of perilymph (PL) were studied 30, 60, and 120 min postmortem. During this period the mean lactate concentration of scala tympani and scala vestibuli increased from 4.8 mmol/l found intravitally to 17.8 and 15.1 mmol/l, respectively, whereas pyruvate decreased from an average of 0.33 to 0.10 mmol/l (fig. 1). These inverse changes of concentration yield postmortem lactate/pyruvate quotients which are more than one order of magnitude higher than the quotients found intravitally (Table 1). In comparative tests of blood samples carried out 30, 60, and 120 min after the sampling (Fig.1), the lactate increase was found to be markedly lower than in postmortem PL. The substantial metabolite changes in PL seem to be caused by glycolytic activity of all cochlear structures that are in direct contact with PL. The decrease of pyruvate level is probably due to a shift of the lactate-pyruvate equilibrium (lactate dehydrogenase system) in PL. The blood vessels in the perilymphatic space can be neglected as postmortem metabolite source of PL.

Animals↗

[On the glucose, pyruvate, and lactate concentration of perilymph, blood, and cerebrospinal fluid of unexposed and sound-exposed guinea pigs under ethyl urethane anesthesia (author's transl)].

Glucose, pyruvate, and lactate of perilymph (PL), blood, and cerebrospinal fluid (CSF) of unexposed and sound-exposed guinea pigs under ethyl urethane anesthesia were examined with due consideration of the principal sources of error. The animals had fasted for 15--20 h before the experiment to stabilize the blood glucose level. The metabolites were determined enzymatically by means of fluorescence measurements. It was found that the glucose levels depend not only on ingestion but also on the duration of anesthesia of the animals before sampling. The mean values of the scala tympani and scala vestibuli PL and CSF did not differ significantly, being about half those of blood or plasma immediately (10--20 min) after introducing anesthesia (Table 2). This concentration difference is in disagreement with the original ultrafiltration hypothesis of PL, suggesting a blood-PL barrier for glucose. The dependence on the duration of anesthesia and on the animals' ingestion before sampling appears to be an important cause of the differences in glucose data published in literature hitherto. No influence of anesthesia on pyruvate and lactate concentrations was observed. Data obtained on unexposed control animals (Tables 3 and 4) confirmed our earlier metabolite findings (Scheibe et al. 1976, 1981). No major changes in glucose, pyruvate, and lactate concentration of PL, blood, and CSF were detectable immediately after 1 h of exposure to wide-band noise at an intensity of 120 dB SPL. The present lactate findings confirmed our earlier exposure experiments (Scheibe et al. 1976), but they did not agree with the information given by Schnieder (1974).

Animals↗

[Lactate and pyruvate concentrations in perilymph, blood, and cerebrospinal fluid of guinea pigs].

Lactate and pyruvate were studied comparatively in perilymph (PL), blood, and cerebrospinal fluid (CSF) of anesthetized guinea pigs. Arterial blood pressure, heart and respiration rate (Fig. 1), and arterial blood-gas state (Table 2) were simultaneously checked in a group of the animals. The metabolites were determined enzymatically by using the fluorometric technique. The studies have shown (Table 1) that both the lactate and the pyruvate concentrations are in PL at a similar rate (about 3:1) higher than in native blood and also higher than in CSF. The metabolite values of blood, especially the lactate values, were lower when blood was taken alone, e.g., more physiological, than in the case when CSF and PL had been sampled before. The lactate/pyruvate ratios of Pl are somewhat higher than the blood ratios. The ratio of CSF was found to be lower. The high metabolite levels in PL suggest an intracochlear origin. A direct perilymphatic lactate origin could not be detected.

Animals↗

Purified human plasma proteins of unknown function.

The development of new analytic and preparative techniques in the field of protein chemistry has essentially extended our knowledge about the variety of human plasma proteins in the last 15 years. In many cases plasma proteins have been particularly determined by immunologic techniques, partially highly purified and physicochemically well characterized, before knowing the biological function. To these proteins belong among others the alpha 1-antitrypsin, Cl-inactivator, alpha 2-macroglobulin, Gc-globulin and the cold-insoluble globulin. Today we know more than 100 proteins being isolated from human plasma and among these are nearly 20, of which the biological function is not yet known. To this group are belonging proteins, which are known since many years, like the alpha 1-acid glycoprotein and the C-reactive proteins as well as proteins, which have only been described in the last years and which partially have an interesting chemical structure, e.g. the histidinerich 3,8S-alpha 2-glycoprotein and the leucine-rich 3,1S-alpha 2-glycoprotein, of which every fifth amino acid is formed by leucine. It is to be hoped that the special chemical structure of some of these human plasma proteins as well as the quantitative immunologic determination in different patient sera will give hints to their biological function.

Alpha-Globulins↗

[Isolation and crystallization of the Clr subunit of the first component of human complement (author's transl)].

Isolation of Clr from Cohn-Fraction I of human plasma is accomplished by a series of steps which include affinity chromatography on IgG-p-azobenzyloxyethylsulfonylethoxy-Sepharose CL-4B, ion exchange chromatography and preparative zone electrophoresis. The protein is easily soluble in 0.1 M Tris/HCl buffer, pH 10.0 and is crystallized readily by means of dialysis against 0.15 M saline, pH 7.0 in the form of hexagonal dipyramides. The structural elucidation using X-ray analysis is now possible.

Chromatography, Affinity↗

Immuno-chemical studies on the alkali-labile carbohydrate chains of human serum glycoproteins.

Human serum glycoproteins can be classified into those containing N-acetyl-D-galactosamine and into those lacking this hexosamine. The N-acetyl-D-galactosamine-containing serum glycoproteins have alkali-labile chains containing this hexosamine linked O-glycosidically to hydroxy amino acids. These alkali-labile chains can be demonstrated in neuraminic acid free serum glycoproteins by gas liquid chromatography and by using precipitating lectins from invertebrates and plants. They are represented by two chains, one containing only N-acetyl-D-galactosamine, the other with D-galactose linked (1--3) beta-glycosidically to this hexosamine forming a disaccharide. Serologically these two chains, which usually occur together on one molecule, can be characterized by their reaction with lectins from Helix pomatia (anti-A like) and from Agaricus bisporus and Arachis hypogaea (anti-TF specificity).

Acetylgalactosamine↗

[Blockage of cochlear aqueduct for examination of perilymph (guinea pig) (author's transl)].

To prevent the perilymph (guinea pig) from contamination with CSF during the sampling the aqueductus cochleae (AC) was blocked by injection of tissue adhesive into the meningeal aperture. The control of an exact blockage of AC was carriedout by examination of perilymph-outflow after opening the cochlea (injection of fluorescein-Na into the CSF-space), analysis of perilymph-protein-concentration, macroscopic and microscopic examination of the temporal bones. In all cochleae we have found the same morphological structures, notwithstanding whether the AC was blocked (for a time from 30 min to 7 weeks) or not: The cochlear aqueduct is filled with a mesh of mesenchymal tissue, which grows more dense towards the cochlear aperture andcontinues into the round window membrane. From scala tympani the AC is always limited by one layer of cells forming a sort of membrane (under light microscope). It seems possible that CSF moves in the inner of the round window membrane between AC and subepithelian space of middle ear mucosa, whereas perilymph of scala tympani is not in direct contact with the flow of CSF. The scala tympanic side of the round window membrane may be a big area for diffusion and there also may be an exchange between CSF and perilymph. The outflow of CSF into the cochlea after experimental opening of the cochlea is an artifact, caused by damage of pressure equilibration between CSF-space and cochlea. 30 min and 5--7 weeks after blockage no morphologicaland electrophysiological alterations from those of the control ears were to be seen. The protein concentration, however, increased significantly 5--7 weeks after blockage from normally about 200 mg/100 ml toalmost the double especially in the scala tympani (see Table 1).

Action Potentials↗