Predominance of a cell population less sensitive to carcinogenesis in neoplastic cells of 3-methylcholanthrene-induced tumors in mouse aggregation chimeras.

Tumors were induced by i.m. injections of 3-methylcholanthrene (0.5 mg) in 100% experimental aggregation chimeras derived from two mouse strains C57Bl/6J (hereafter called B6) and A/J, dimorphic for the enzyme glucosephosphate isomerase (Gpi-1a or Gpi-1b) and differing in coat color, aryl hydrocarbon hydroxylase inducibility, and cytotoxic activities of natural killer cells and macrophages. In the majority of host tissues and organs, such as coat, lung, spleen, and skeletal muscle, the B6 phenotype was predominant. Likewise, the nonneoplastic intratumoral host cells of both induced primary tumors and parental tumor transplants in chimeras were of B6 origin. In contrast, neoplastic cells in 70% of the tumors originated exclusively from the less aryl hydrocarbon hydroxylase-inducible and less immune competent A/J strain. The A/J origin was verified by subsequent cell culturing of the tumors. Only 30% of the tumors contained neoplastic cells of both A/J and B6 phenotype. A further reduction of mixed tumors was achieved with lower doses (0.1 and 0.06 mg) of the carcinogen. The dominance of the A/J phenotype of the tumors contrasted not only with aryl hydrocarbon hydroxylase inducibility and host cell composition but also with tumor pathogenesis (at 0.5 mg 3-methylcholanthrene) in the parental strain. Whereas tumor incidence was 100% in the B6 strain, it was only 65% in A/J mice, with tumors also developing later. As the in vivo and in vitro growth rates of parental strain-derived tumors were comparable, a cell selection caused by different growth rates favoring the A/J phenotype appeared unlikely.

Nuclear columns, Kinetics of RNA synthesis and release in isolated rat liver nuclei.

By continuous perfusion of columns containing isolated immobilized rat liver nuclei with media containing labeled RNA precursors, the in vitro synthesis and release of RNA was studied. The combined reaction of synthesis and release could be adjusted to proceed at a constant rate. The reaction rate responded to variation of termperature, ionic conditions, nucleoside triphosphate concentration and to the addition of RNA polymerase inhibitors. During 60 min perfusion approximately equal amounts of radioactive low molecular weight RNA and of ribonucleoproteins were released. Pulse-chase experiments showed that the low molecular weight RNA was synthesized throughout the perfusion and released immediately after formation. The ribonucleoproteins were primarly labeled during the first period of perfusion and were gradually released. Synthesis of RNA contained in the ribonucleoproteins was inhibited by low alpha-amanitin concentrations, indicating that it was catalyzed by RNA polymerase II. The in vitro labeled ribonucleoproteins exhibited properties of the stable nuclear particles which can be extracted from isolated nuclei after rapid in vivo labeling of RNA. They had a buoyant density of 1.41--1.43 in CsCl, were partially unstable in 1% deoxycholate, but stable in 0.1% deoxycholate, in 100 mM NaCl and in 10 mM EDTA. Due to the dilution by the perfusion medium, the ribonucleoproteins sedimented with a peak at 22--27 S, and not at 30--45 S. The RNA synthesized in the immobilized nuclei was not degraded during the perfusion. Less than 20% was gradually released, whereby the 20--30 S peak zone was reduced. While the properties of the in vitro labeled ribonucleoproteins and of rapidly in vivo labeled ribonucleoproteins were the same, the kinetics of their release differed.

Proteome analysis of nuclear matrix proteins during apoptotic chromatin condensation.

The nuclear matrix (NM) is considered a proteinaceous scaffold spatially organizing the interphase nucleus, the integrity of which is affected during apoptosis. Caspase-mediated degradation of NM proteins, such as nuclear lamins, precedes apoptotic chromatin condensation (ACC). Nevertheless, other NM proteins remain unaffected, which most likely maintain a remaining nuclear structure devoid of chromatin. We, therefore, screened various types of apoptotic cells for changes of the nuclear matrix proteome during the process of apoptotic ACC. Expectedly, we observed fundamental alterations of known chromatin-associated proteins, comprising both degradation and translocation to the cytosol. Importantly, a consistent set of abundant NM proteins, some (e.g. hNMP 200) of which displaying structural features, remained unaffected during apoptosis and might therefore represent constituents of an elementary scaffold. In addition, proteins involved in DNA replication and DNA repair were found accumulated in the NM fraction before cells became irreversibly committed to ACC, a time point characterized in detail by inhibitor studies with orthovanadate. In general, protein alterations of a consistent set of NM proteins (67 of which were identified), were reproducibly detectable in Fas-induced Jurkat cells, in UV-light treated U937 cells and also in staurosporine-treated HeLa cells. Our data indicate that substantial alterations of proteins linking chromatin to an elementary nuclear protein scaffold might play an intriguing role for the process of ACC.

A novel spliced transcript of human CLAPS2 encoding a protein alternative to clathrin adaptor protein AP17.

Transcripts of genes encoding proteins of clathrin complexes have been reported to undergo tissue-specific alternative splicing. AP17, encoded by human CLAPS2 cDNA, is the small chain of the major clathrin adaptor complex AP-2 associated with mammalian plasma membranes. In this study, two cDNAs were isolated from a cDNA library of human blood cells. Whereas one cDNA encoded AP17, the other cDNA encoded a putative novel protein variant, termed AP17Delta. Both coding regions were completely sequenced. Consisting of 142aa residues, the predicted protein AP17Delta of 12kDa lacks 38aa residues of AP17. Using specific primers for RT-PCR, mRNAs for AP17Delta and AP17 were found in leukocytes and cultured leukemia cells. The finding of a putative intron in a human EST cDNA clone suggests that mRNAs for AP17 and AP17Delta are formed by alternative splicing. In addition, the identity of human and rat AP17 amino acid sequences is demonstrated.

Elevated plasma levels of crosslinked fibrinogen gamma-chain dimer indicate cancer-related fibrin deposition and fibrinolysis.

Cancer-related fibrin deposition and fibrinolysis were investigated by two-dimensional gel electrophoresis of human solid tumor and effusion specimen in addition to plasma samples. Fibrinogen gamma-chain dimer indicating fibrin deposition and plasmin-generated fibrinogen beta-chain fragments were identified in various solid tumor types by amino acid sequencing, mass spectrometry analysis and Western blotting. In tumor-associated effusions, these techniques allowed to observe plasmin-generated fragments of fibrinogen alpha, beta and gamma-chains in addition to elevated levels of acute-phase proteins. Similar observations were made in case of inflammation-associated effusions. No fibrin degradation product was observed in plasma samples, however, high amounts of fibrinogen gamma-chain dimer crosslinked by transglutaminase were detected in plasma from tumor patients, but not in plasma from controls and patients suffering acute infections and/or inflammations. This finding demonstrated that high transglutaminase activity may be associated with cancer. The presented data indicate that the amount of crosslinked fibrinogen gamma-chain dimer in plasma may correlate with tumor-associated fibrin deposition. The tumor-biological relevance of this potential marker protein is discussed.

Autoantibodies in chronic GVHD: high prevalence of antinucleolar antibodies.

Sera from 32 bone marrow allograft recipients were screened for the presence of autoantibodies 4-61 months post-graft. Sera from 12 of 19 patients with extensive chronic graft-versus-host disease (c-GVHD) stained the nucleolar region strongly in immunofluorescence, indicating the presence of specific antinucleolar antibodies. In contrast, none of three patients with limited and none of 10 patients without c-GVHD had antinucleolar antibodies. Antibodies reacting with nuclear constituents other than nucleoli were found in five of the 12 antinucleolar positive patients. The appearance of antinucleolar antibodies coincided with early clinical symptoms of c-GVHD. We conclude that the appearance of antinucleolar antibodies after bone marrow transplantation is specific for patients with extensive c-GVHD. Furthermore, the development of extensive c-GVHD is paralleled by the emergence of these antinucleolar antibodies.

Continuum saltation model for sand dunes.

We derive a phenomenological continuum saltation model for aeolian sand transport that can serve as an efficient tool for geomorphological applications. The coupled differential equations for the average density and velocity of sand in the saltation layer reproduce both the known equilibrium relations for the sand flux and the time evolution of the sand flux as predicted by microscopic saltation models. The three phenomenological parameters of the model are a reference height for the grain-air interaction, an effective restitution coefficient for the grain-bed interaction, and a multiplication factor characterizing the chain reaction caused by the impacts leading to a typical time or length scale of the saturation transients. We determine the values of these parameters by comparing our model with wind tunnel measurements. Our main interest are out of equilibrium situations where saturation transients are important, for instance at phase boundaries (ground/sand) or under unsteady wind conditions. We point out that saturation transients are indispensable for a proper description of sand flux over structured terrain, by applying the model to the windward side of an isolated dune, thereby resolving recently reported discrepancies between field measurements and theoretical predictions.

Coenzyme Q10, a cutaneous antioxidant and energizer.

The processes of aging and photoaging are associated with an increase in cellular oxidation. This may be in part due to a decline in the levels of the endogenous cellular antioxidant coenzyme Q10 (ubiquinone, CoQ10). Therefore, we have investigated whether topical application of CoQ10 has the beneficial effect of preventing photoaging. We were able to demonstrate that CoQ10 penetrated into the viable layers of the epidermis and reduce the level of oxidation measured by weak photon emission. Furthermore, a reduction in wrinkle depth following CoQ10 application was also shown. CoQ10 was determined to be effective against UVA mediated oxidative stress in human keratinocytes in terms of thiol depletion, activation of specific phosphotyrosine kinases and prevention of oxidative DNA damage. CoQ10 was also able to significantly suppress the expression of collagenase in human dermal fibroblasts following UVA irradiation. These results indicate that CoQ10 has the efficacy to prevent many of the detrimental effects of photoaging.

[Relation between yoga and psychotherapy]. / Zum Verhältnis von Yoga und Psychotherapie.

Western psychotherapy and Yoga overlap insofar as both systems are based upon religious and mythological facts, and--on the level of psychotherapeutical praxis--on the technics of hypnosis, auto- and heterosuggestions and/or meditation. It is 50 years ago that the west considered the psychotherapeutical effects of theayoga-systems, first of all of Hatha- Yoga. Even today a theoretical foundation by means of proper comparison of the two structures is missing. In present-day India Yoga fulfills psychohygienical functions without being a psychotherapy in our sense. There are various techniques of magic in use, which replace the experimental psychological-psychotherapeutical methods in the West. The acceptance of meditation could only be successful if the metaphysical and sociocultural context would be integrated at the same time. The traditional function of Yoga guarantees its continuity even in the modern industrialized society of India. Neither the theoretical nor the pratical fundaments allow a direct transfer at present.

Studies on ribonucleic acid metabolism using nuclear columns. Release of rapidly labeled RNA from rat liver nuclei.

A method is described to study the effect of successively changing incubation conditions on the release of rapidly labeled RNA from isolated nuclei. Nuclear columns containing immobilized rat liver nuclei isolated after in vivo application of labeled orotic acid are perfused with different non-radioactive media. Within the course of one perfusion, the rate of RNA release can be repeatedly altered by variation of temperature, acidity and concentrations of nucleoside triphosphates, complexing agents, sodium chloride and manganese chloride. RNA release can be started and stopped, indicating that the reaction does not result from damage to nuclei. During 60 min perfusion the same product, labeled ribonucleoprotein (sigma = 1.43 g/cm3 in CsCl), is released. High release rates depend on the ratio of nucleoside triphosphate to divalent cation concentration, not on the concentration of the agents per se. Ribonucleoside and deoxyribonucleoside triphosphates exert the same effect as ATP. The SH reagents iodoacetamide and iodoacetate only slightly affect the ATP-induced reaction. In contrast, p-chloromercuribenzoate, after an initial stimulation, causes inhibition of RNA release.

[Modern cosmetic agents and evidence of their function]. / Moderne kosmetische Wirkstoffe und der Nachweis ihrer Funktion.

Fluorometers modified by quartz light conductors allow the recording of the skin-intrinsic fluorescence. Signals assigned to the aromatic amino acids permit a prognosis concerning the sensitivity of untreated skin to light and the sunscreen factor to be expected after the application of UV filters. We discuss the microdistribution and skin penetration of UV filters and describe the utilization of the image analysis technique both for the count and size determination of fluorescent sebaceous glands and for the quantification of the effects of antiperspirants. Based both on the selective increase of efficacy of deodorant soaps, analytically controlled, and on the differences of the deodorizing effects found in various test groups, we discuss the possibilities of improving deodorants.

Nuclear matrix proteins specific for subtypes of human hematopoietic cells.

Nuclear matrices were prepared from isolated subtypes of human hematopoietic cells and from cultured leukemia cells. The nuclear matrix proteins were analyzed by high-resolution two-dimensional gel electrophoresis and computer-assisted image analysis. While more than 200 protein spots were shared among the cells, about 50 distinct spots were found characteristic for individual cells or groups of related cells. This allowed to differentiate between hematopoietic cells and nonhematopoietic cells, lymphocytes and myeloid cells, monocytes, neutrophils, and promyelocytic leukemia cells. B and T lymphocytes could not be differentiated. Myeloid cells with their polymorph nuclei were characterized by the presence of 13 and by the absence of seven distinct spots, as well as by low concentrations of nuclear lamins and of heterogeneous nuclear ribonucleoproteins. Neutrophils with multilobular nuclei displayed six additional spots, while lacking 18 nuclear matrix protein spots. The nuclear matrix of proliferating cells showed three distinct spots in addition to proliferating cell nuclear antigen, increased concentrations of numatrin (B23), and heterogeneous nuclear ribonucleoproteins. The described cell-specific nuclear matrix proteins may represent new markers for hematopoietic cells.