HuminsÄure und PermeabilitÄt

Ohne Zusammenfassung     

Über die Alterung des neurosekretorischen Zwischenhirn-Systems in Zusammenhang mit der Regulation des Wasserhaushaltes

Zusammenfassung Der Alterungsprozeß der neurosekretorischen hypothalamo-neurohypophysären Regulationszentren des Wasserhaushaltes wurde an einem großen Kollektiv weißer Laboratoriumsratten verfolgt. Es konnte nachgewiesen werden, daß die Neurosekret- und damit die Hormonproduktion bis zum hohen Alter erhalten bleiben. Erst sehr alte Tiere zeigen eine mehr oder weniger deutliche Einschränkung der Neurosekretproduktion. Die Befunde werden mit den aus der Literatur bekannten Beobachtungen über die altersbedingten Veränderungen der Nieren in Zusammenhang gebracht. Wie während des Wachstums der Reifezustand der Niere auf allen Entwicklungsstufen der Reife der Hormonproduktionsstätten entspricht und beide wiederum mit den mit zunehmendem Alter sich weiter differenzierenden Erfordernissen des Allgemeinstoffwechsels koordiniert sind, so befinden sich auch im gesunden alternden Organismus neurosekretorisches Regulationszentrum, peripheres Erfolgsorgan (distaler Nierentubulus) und die mit dem Wasserhaushalt in Verbindung stehenden Stoffwechselprozesse dauernd im Zustand einer wohlausgewogenen Harmonie. Ein unkoordinierter Abbau eines Gliedes dieser Kette muß zwangsläufig zu Fehlregulation und Krankheit führen.Durchgeführt mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Das Subfornikalorgan und seine Beziehungen zu dem neurosekretorischen System im Zwischenhirn des Frosches

Zusammenfassung Das Subfornikalorgan von Rana esculenta und Rana temporaria liegt am Zusammenfluß dreier Ventrikel in der Pars ventromedialis oder septalis des Telencephalon und weist einen bei Säugetieren nicht erkennbaren Bauplan in drei Zonen oder Schichten auf. Die innere Zone wird von einem glomerulumartigen Gefäßsinus mit perivaskulärem Raum dargestellt. Große, nur von Gliamembranen getrennte Vakuolen umgeben als mittlere Zone das Gefäß. Diese Schicht ist praktisch zellfrei. Die äußere Schicht wird im ventrikulären Bereich von sehr unterschiedlich gebauten Ependymzellen gebildet. Sie können hochprismatisch bis endothelartig platt sein. Die anderen dem Gehirn zugewandten Seiten der dritten Zone bestehen aus Gliazellen, unter denen drei Zellarten gefunden werden, die keine Ähnlichkeit mit den Parenchymzellen der Säugetiere haben. Im basalen Bereich kommen Zellen vor, deren Cytoplasma sich mit Chromhämatoxylin und Aldehydthionin tingiert und die faserige Fortsätze bilden. Auch im Ependym und zwischen den Vakuolen werden in Einzelfällen Gomori-positive Substanzen gefunden.Durch osmotische Belastung und Hypophysektomie der Tiere wurde versucht, Bahnen zwischen Nucleus praeopticus und Subfornikalorgan darzustellen. Es konnte gezeigt werden, daß zwischen beiden Bezirken des Gehirns eine Verbindung besteht, deren Hauptweg über den Commissurenwulst der Commissura anterior und Commissura pallii anterior zum Subfornikalorgan führt. Unter experimentellen Bedingungen ließen sich auch die im Normalfall nur selten vorkommenden Gomori-positiven Substanzen im Ependym und zwischen den Vakuolen regelmäßiger nachweisen.Der Drei-Schichten-Bau, in dem sich die Flüssigkeitssysteme Blut und Liquor unter Vermittlung eines dritten — dem Vakuoleninhalt — gegenüberstehen, und die Verbindung zum neurosekretorischen System des Zwischenhirns werden für die Funktion des Organs als bedeutsam erachtet.     

Purification and Characterization of Sorbitol Dehydrogenase from Bovine Brain

Sorbitol dehydrogenase (EC 1.1.1.14) was isolated from bovine brain and purified 3,000-fold to apparent homogeneity, as judged by polyacrylamide gel electrophoresis. The purified enzyme had a specific activity of 36 units/mg of protein; a molecular weight of 39,000 for each of the four identical subunits and 155,000 for the intact enzyme were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel exclusion chromatography, respectively. The presence of one Zn2+ per subunit was confirmed by atom absorption spectroscopy; inactivation of the enzyme by metal-chelating agents points to the essential role that Zn2+ plays in the catalytically competent enzyme. The enzyme is also inactivated by thiol-blocking reagents; with respect to inactivation by sodium pyrophosphate, sorbitol dehydrogenase is different from closely related alcohol dehydrogenase.     

Iron overload of the liver by trimethylhexanoylferrocene in rats.

Iron-deficient female Wistar rats were fed a diet, which contained 0.5% trimethylhexanoylferrocene, over a 56-week period. This dietary iron loading resulted in a progressive siderosis and enlargement of the liver with a maximum iron content of 947.0 +/- 148.0 mg (vs. 0.07 +/- 0.04 mg in iron deficiency) and a maximum organ weight of 39.4 +/- 6.6 g (vs. 6.9 +/- 1.4 g in iron-deficient control rats). Up to 43 weeks, whole liver iron rose by increase in iron concentration (max. 28.0 +/- 6.1 mg/g wet weight, w.w.) as well as by enlargement of the organ. Afterwards whole liver iron increased solely by ongoing hepatomegaly. At the commencement of iron loading, stainable iron was almost exclusively stored by hepatocytes equally throughout all areas of the liver lobule. Later, the distribution of iron-loaded hepatocytes became strikingly periportal, and, in addition, Kupffer cells as well as sinus-lining endothelia began to store iron. Animals with a liver iron concentration of more than 10.4 +/- 0.75 mg/g w.w. showed no further increase in ferritin and haemosiderin within hepatocytes. Iron-burdened Kupffer cells/macrophages, however, accumulated permanently, hereby forming intrasinusoidal and portal siderotic nodules and areas. First signs of liver damage such as necrosis of single hepatocytes and mild fibrosis began at a liver iron concentration of 14.7 +/- 1.4 mg/g w.w. With advancement of iron loading, focal necrosis of hepatocytes and iron-burdened macrophages took place, and significant perisinusoidal as well as portal fibrosis developed. Cirrhosis, however, the final stage of impairment in iron overload of the liver in humans, could not be induced in this animal model up to now.     

Protein kinase C activity is rate limiting for shedding of the interleukin-6 receptor.

An analysis of the mechanism of generation of the soluble interleukin-6 receptor (IL-6R) has been performed. The membrane-bound receptor is proteolytically cleaved to release a soluble receptor form which retained its ligand binding capacity. Furthermore, the soluble IL-6R is unique in its ability to induce a biological signal in complex with the ligand interleukin-6 (IL-6) on cells which by themselves do not bind IL-6. Shedding of the IL-6R is strongly activated by PMA and can be inhibited by the protein kinase inhibitor staurosporine. The generation of the IL-6R is not dependent on protein synthesis. The inactive PMA analogue 4-alpha-phorbol-12,13-didecanoate fails to induce shedding of the IL-6R. Transfection of a protein kinase C expression plasmid into IL-6R expressing cells leads to enhanced shedding of the receptor. These experiments clearly show that protein kinase C regulates shedding of the IL-6R.     

A quantitative assay for tyrosine sulfation and tyrosine phosphorylation in peptides.

A method was developed to measure sulfation and phosphorylation of tyrosine in proteins after alkaline hydrolysis, ion-exchange chromatography, reaction with [3H]dinitrofluorobenzene and subsequent thin-layer chromatography. The method allows the detection of 10-20 pmol of modified tyrosine and was applied to determine the content of tyrosine-phosphate and -sulfate in fibrinogens, thyroglobulin, alpha-casein, cytochrome c and glyceraldehyde dehydrogenase.     

An Experimental Test of the Rhizopine Concept in Rhizobium meliloti

In some Rhizobium-legume symbioses, compounds known as rhizopines are synthesized by bacteroids and subsequently catabolized by free-living cells of the producing strain. It has been suggested than rhizopines act as proprietary growth substrates and enhance the competitive ability of the producing strain in its interactions with the diverse microbial community found within the rhizosphere. Wild-type, rhizopine-producing Rhizobium meliloti L5-30 and mutant L5-30 strains deficient for either rhizopine synthesis or catabolism were inoculated onto lucerne host plants in competition experiments. These experiments demonstrated that no apparent advantage resulted from the ability to synthesize a rhizopine, whereas the ability to catabolize rhizopine provided a clear advantage when an organism was in competition with a strain without this ability. The results suggest that when an organism is in competition with a catabolism-deficient mutant, the ability to catabolize rhizopine results in enhanced rates of nodulation. The results of the experiments were not consistent with the hypothesis that the sole role of rhizopines is to act as proprietary growth substrates for the free-living population of the producing strain.     

Line-scanning microphotolysis for diffraction-limited measurements of lateral diffusion.

Fluorescence microphotolysis was combined with confocal laser-scanning microscopy to yield a method, herein referred to as line-scanning microphotolysis (LINESCAMP), for the measurement of molecular transport at a lateral resolution of approximately 0.34 microns and a temporal resolution of approximately 0.5 ms. A confocal microscope was operated in the line scan mode, while the laser beam power could be switched during scanning between low monitoring and high photolysing levels in less then a microsecond. The number and location of line segments to be photolysed could be freely determined. The length of the photolysed segments could be also chosen and was only limited by diffraction. Together with instrumentation a new, completely general, theoretical framework for the evaluation of diffusion measurements was developed. Based on the numerical simulation of diffusion processes employing a modified Crank-Nicholson scheme, the theory could be applied to any photobleaching geometry and profile as the initial condition and took into account the convolution with the microscope point spread function. With small diffraction-limited areas, the method yielded accurate values for diffusion coefficients in the range between approximately 10(-4) and 1 micron2 s-1. A first application of the method to the diffusion of a fluorescently labeled tracer inside the cell nucleus showed the potential of the method for the study of complex biological systems.