Die Intravitale Karyorrhexis Der Exokrinen Pankreaszelle im Elektronenmikroskopischen Bild

Zusammenfassung Die intravitale Karyorrhexig wird am exokrinen Pankreas der Maus nach Osmium- und Formalinfixierung licht- und elektronenmikroskopisch untersucht. Im Verlaufe der Karyorrhexis kommt es zu einer Trennung des Karyoplasmas in 'Chromatin und 'Karyolymphe (Interchromatin-Substanz). In fortgeschrittenen Stadien ist das Bild solcher Zellkerne — im Gegensatz zu normalen Intermitosekernen — nach beiden Fixierungsmethoden fast identisch. Das Chromatin ist in dichten, membranständigen Kappen konzentriert. Der übrige Kernraum wird — abgesehen vom Nukleolus — von der hellen Karyolymphe ausgefüllt. In ihrem Bereich treten die sog. Interchromatingranula (200–250 Å), einzeln oder in dichten Haufen, meist deutlich hervor. Der Nukleolus, dessen Feinstruktur nicht auffallend verändert ist, liegt entweder frei oder den Chromatinkappen unmittelbar an. Die Kernhülle ist bei fortgeschrittener Karyoplasmaentmischung unversehrt oder über kürzere oder längere Strecken geschwunden. Im Endstadium der Karyorrhexis sind mehrere, von Kernmembranfragmenten unvollständig begrenzte Chromatinbrocken wahllos im Zelleib verstreut. Die cytoplasmatischen Strukturen können auch bei ausgeprägten Kernveränderungen noch weitgehend normal aussehen.Die karyorrhektischen Kernveränderungen werden als Folge einer 'Entmischung des Karyoplasmas gedeutet, für die eine Dehydratation der DNS-tragenden Chromosomenstrukturen verantwortlich gemacht wird. Auch das Sichtbarwerden und das Verklumpen der 'Interchromatingranula wird auf eine Dehydratation zurückgeführt.

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Summary The intravital caryorrhexis in the exocrine pancreas of the mouse was studied by means of light- and electron microscopy after fixation with osmium and formaldehyde. During caryorrhexis the caryoplasm segregates into chromatin and caryolymphe (interchromatinic substance). In contrast to normal intermitotic nuclei both fixations result in a very similar picture of the typical caryorrhectic nuclei. The chromatin is concentrated in dense caps associated with the nuclear envelope. Except those dense caps and apart from the nucleolus the whole nucleus is occupied by the light caryolymphe. In this area the so-called interchromatinic granules (200–250 Å) become clearly visible, isolated or in dense clumps. The nucleolus, the fine structure of which has not remarkably changed, can be found either isolated or in touch with the chromatin caps. During advanced segregation of the caryoplasm the nuclear envelope was found to be either unimpaired or had disappeared for variable lenghths. In the final stage of caryorrhexis several chromatin clumps — partly confined by fragments of the nuclear envelope — are distributed irregularly within the cell. Even in the case of pronounced nuclear changes the cytoplasmic structures may appear almost unaffected.It is assumed that the nuclear changes during caryorrhexis are due to a segregation of the caryoplasm, caused by dehydration of the DNA-carrying chromosome fibrils. Also dehydration is made responsible for the appearance and clumping of the interchromatinic granules.

     

Determination of amino sugars and amino acids in glycoconjugates using precolumn derivatization with o-phthalaldehyde.

This report examines the RP-HPLC separation of o-phthalaldehyde derivatives of amino acids, amino sugars, and amino sugar alcohols using either 2-mercaptoethanol or 3-mercaptopropionic acid. A method with pmol sensitivity for the analysis of N-acetylamino sugars of glycoconjugates was elaborated. Upon hydrolysis, amino sugars are reduced with borohydride. Automated precolumn derivatization and chromatographic conditions for the resulting hexosaminitols are the same as those used for the analysis of amino acids. The method has been tested with as little as 2 micrograms of bovine fetuin, with a glycopeptide from bromelain and with an oligosaccharide after periodate oxidation.     

Chemical structure of the lipopolysaccharide of Haemophilus influenzae strain I-69 Rd-/b+. Description of a novel deep-rough chemotype

The chemical structure of the lipopolysaccharide of a deep-rough mutant (strain I-69 Rd-/b+) of Haemophilus influenzae was investigated. The hydrophilic backbone of lipid A was shown to consist of a beta-(1',6)-linked D-glucosamine disaccharide with phosphate groups at C-1 of the reducing D-glucosamine and at C-4' of the non-reducing one. Four molecules of (R)-3-hydroxytetradecanoic acid were found directly linked to the lipid A backbone, two by amide and two by ester linkage (positions 2,2' and 3,3', respectively). Laser-desorption mass spectrometry showed that both 3-hydroxytetradecanoic acids linked to the non-reducing glucosamine carry tetradecanoic acid at their 3-hydroxyl group, so that altogether six molecules of fatty acid are present in lipid A. The lipopolysaccharide was the first described to contain only one sugar unit linked to lipid A. This, sugar in accordance with a previous report [Zamze et al. (1987) Biochem. J. 245, 583-587], was shown to be a dOclA phosphate. The phosphate group was found at position 4, but the analytical procedures employed (permethylation and methanolysis followed by gas-liquid chromatography/mass spectrometry) also revealed dOclA 5-phosphate. Since a cyclic 4,5-phosphate could be ruled out by 31P-NMR, we conclude that, in this lipopolysaccharide, a mixture of dOclA 4- and 5-phosphate is present. By methylation analysis of the dephosphorylated, deacylated and reduced lipopolysaccharide the attachment site of the dOclA was assigned to position C-6' of the non-reducing glucosamine of lipid A. The anomeric linkages present in the lipopolysaccharide were assessed by 1H-NMR and 13C-NMR of deacylated lipopolysaccharide. The saccharide backbone of this Haemophilus influenzae lipopolysaccharide possesses the following structure: (Formula; see text)     

Single proline substitutions in predicted alpha-helices of murine granulocyte-macrophage colony-stimulating factor result in a loss in bioactivity and altered glycosylation

Contributions of alpha-helices to biological activity in murine granulocyte-macrophage colony-stimulating factor were analyzed using site-directed mutagenesis and protein expression in COS-1 cells. A series of single proline substitutions were made for residues within the four predicted alpha-helices as a means of disrupting local helical secondary structure. Mutations in three of the four helices resulted in marked reductions in bioactivity. Five mutants E21P, L56P, E60P, L63P, and L107P showed 10(2)-10(4)-fold reduction in bioactivity as well as hyperglycosylation. The same Pro substitutions made on non-N-glycosylated molecules had a similar loss in bioactivity implying that a Pro-induced structural change and not hyperglycosylation was responsible for the major decrease in bioactivity. Additional amino acid substitutions at these residues which conserved charge or hydrophobicity, or replaced the original residue with an Ala, verified that conformational changes in the protein structure were specifically due to steric constraints imposed by the Pro residue rather than loss of important side chain functions.     

Predictability of Extreme Events in Social Media

It is part of our daily social-media experience that seemingly ordinary items (videos, news, publications, etc.) unexpectedly gain an enormous amount of attention. Here we investigate how unexpected these extreme events are. We propose a method that, given some information on the items, quantifies the predictability of events, i.e., the potential of identifying in advance the most successful items. Applying this method to different data, ranging from views in YouTube videos to posts in Usenet discussion groups, we invariantly find that the predictability increases for the most extreme events. This indicates that, despite the inherently stochastic collective dynamics of users, efficient prediction is possible for the most successful items.     

Changes in 20S proteasome activity during ageing of the LOU rat

Muscular functions decline and muscle mass decreases during ageing. In the rat, there is a 27% decrease in muscle protein between 18 and 34 months of age. We examined age-related changes in the proteasome-dependent proteolytic pathway in rats at 4, 18, 24, 29 and 34 months of age. The three best characterised activities of the proteasome (chymotrypsin-like, trypsin-like and peptidylglutamyl peptide hydrolase) increased to 29 months and then decreased in the senescent animal. These variations in activity were accompanied by an identical change in the quantity of 20S proteasome measured by Western blot, whereas the S4 subunit of the 19S regulator and the quantity of ubiquitin-linked proteins remained constant. mRNA of subunits C3, C5, C9, and S4 increased in the senescent animal, but ubiquitin mRNA levels were unchanged. These findings suggest that the 20S proteasome may be partly responsible for the muscular atrophy observed during ageing in the rat.     

T cell alloresponses against HLA-DQ and -DR products involve multiple epitopes on the CD4 molecule. Distinct mechanisms contribute to the inhibition of HLA class II-dependent and -independent T cell responses by antibodies to CD4.

The involvement of the human CD4 molecule in T cell alloresponses to transfected HLA-DQ and -DR Ag was investigated with antibodies to defined CD4 epitopes. Anti-CD4 reagents inhibited T cell proliferation in a dose-dependent manner, and affected responses to HLA-DQ and -DR products equally well. As previously observed for conventional alloresponses, saturating concentrations of CD4 antibodies were required (and sufficient) for substantial blocking of T cell responses to transfected HLA-DQ and -DR products, and antibodies to a wide range of CD4 epitopes were inhibitory. In contrast to these results on class II-dependent T cell proliferation, MHC-independent T cell activation (via CD3 antibodies) was largely resistant to inhibition with the same dose range of CD4 mAb (provided that CD3 and CD4 reagents could not compete for the same class of FcR). This observation validates our conclusion that despite the substantial sequence differences between HLA-DQ and -DR heterodimers, the participation of CD4 epitopes in T cell responses to these molecules is conserved.     

Body size and fatness of free-living baboons reflect food availability and activity levels

We used morphometric techniques and isotope-labeled water to investigate the influence of abundant, accessible food and resultant low activity levels on body size and fatness in free-living adolescent and adult baboons as compared to animals in the same population that experienced more typical, wild-feeding conditions. Females that had access to abundant food from a nearby garbage dump averaged 16.7 kg body mass, 50% more than their wild-feeding counterparts in adjacent home ranges. Little of the difference was due to lean mass: the animals with an accessible abundance of food averaged 23.2% body fat in contrast to 1.9% for the wild-feeding animals. Significant differences between feeding conditions were found for all measured skinfolds and for upper arm circumference but not for linear measurements. Differences between feeding conditions were less for males than for females, perhaps reflecting persistent effects of nutritional conditions during the first eight years of life before dispersal from the group of birth. The difference in fatness between feeding conditions was similar to the difference between humans with frank obesity and those that are considered lean, but in both cases the percentages of body fat in the baboons were considerably less than those observed in humans. In levels of fatness, the relatively sedentary animals resembled their counterparts in group-housed captive conditions. © 1993 Wiley-Liss, Inc.