DNA-damaging properties and cytotoxicity in human fibroblasts of tetrachlorohydroquinone, a pentachlorophenol metabolite

The DNA-damaging potential of pentachlorophenol (PCP) and its metabolite tetrachlorohydroquinone (TCH) was investigated. TCH was found to bind covalently to calf-thymus DNA and to cause single-strand breaks in PM2 DNA. No DNA-damaging effects were observed for PCP. Exposure of human fibroblasts to PCP and TCH showed that TCH is more toxic, when colony-forming ability after exposure to the agent is used as a measure of toxicity. In the evaluation of the mutagenic and carcinogenic potential of PCP the metabolite TCH should be taken into consideration.     

Functional implications of oligomerization of simian virus 40 large T antigen during lytic virus infection.

The formation of oligomers of simian virus 40 (SV40) large T antigen in SV40-infected and -transformed monkey cells was analyzed by sucrose density gradient centrifugation. The overall distribution of total T antigen during lytic infection showed mainly low-molecular-weight forms (monomers and dimers) in the early phase (10 h postinfection) and an increase in the number of oligomers in the late phase of the lytic cycle (36 h postinfection), indicating an accumulation of these final products. In contrast, studying the conversion of newly synthesized T antigen into oligomers by appropriate pulse-chase radiolabeling of infected cells revealed that this processing decelerates considerably during the late phase of infection. This mechanism can be reaccelerated by blocking DNA replication with aphidicolin. Since none of these results could be obtained by using synchronized SV40-transformed monkey cells (COS-1), these observations are compatible with the idea that the process of T antigen oligomerization may be involved in viral, but not in cellular, DNA synthesis.     

Ultrastructural localization of the calcium-binding protein parvalbumin in neurons of the song system of the zebra finch,Poephila guttata

Summary The distribution of parvalbumin (PV) within neurons of the vocal motor nucleus hyperstriatum ventralepars caudalis (HVc) was investigated in the forebrain of adult male zebra finches by means of light and electron microscopy using the indirect immunoperoxidase technique. Parvalbumin-reaction product was located in the amorphous material of perikarya, dendrites and nuclei, and associated to microtubuli, postsynaptic densities and intracellular membranes; it was found in some axons and Gray type-2 boutons, but rarely in type-1 boutons and never in the Golgi apparatus. These observations suggest that parvalbumin may regulate calcium-dependent processes at the postsynaptic membrane and in the cytosol. Furthermore, the partial association of parvalbumin to microtubuli points to an involvement in calcium-dependent tubular functions. Calcium currents and microtubular assembly or transport may be relevant for the known functions of HVc in song learning.     

Morphometrisch-ultrastrukturelle Untersuchungen am Nucleus supraopticus der Ratte nach Dehydratation

Zusammenfassung Der Nucleus supraopticus der Ratte, die einer Dehydratation ausgesetzt war, wurde ultrastrukturell-morphometrisch analysiert. Dabei zeigte sich, daß die relativen Volumenanteile der einzelnen Zellkompartimente während der fünftägigen Durstperiode eine auffallende Konstanz aufweisen. Hingegen läßt sich eine absolute Zunahme der Einzelzellvolumina und somit auch der an der Synthese und Sekretion der Neurohormone beteiligten Zellkompartimente feststellen. Die vorliegenden Befunde sprechen für einen beschleunigten Abtransport des neurosekretorischen Materials bei gesteigerter Synthese. Auf eine optimale Standardisierung der Perfusionsmethode bei Untersuchungen am neurosekretorischen Zwischenhirnsystem wird hingewiesen.

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Morphometric-ultrastructural investigations on the supraoptic nucleus of dehydrated rats

Summary The supraoptic nucleus of the dehydrated rat has been analysed by electron microscopy and morphometry. With that it appears, that the relative volumes of the different cell compartments are striking constant. Otherwise one can see an absolute increase of the cell volume together with the cell compartments which take part at the synthesis and secretion of the neurohormones. These results are expression of an accelerated move of the neurosecretory material during increased synthesis. The importance of an optimal standardization of the perfusion-method in investigations of the neurosecretory system is demonstrated.

Wesentliche Teile der vorliegenden Arbeit werden von H. F. Reinhardt der Medizinischen Fakultät der Universität Basel als Dissertation vorgelegt.     

Restoration of membrane incorporation of an Escherichia coli outer membrane protein (OmpA) defective in membrane insertion

The mechanism of sorting, to the outer membrane, of the 325-residue Escherichia coli protein OmpA has been investigated. It is thought to traverse the membrane eight times in antiparallel beta-strands, forming an amphiphilic beta-barrel which encompasses residues 1 to about 170; the COOH-terminal moiety is periplasmic. A mutant, carrying the substitutions Leu164----Pro and Val166----Asp within the last beta-strand (residues 160-170), has been described which was unable to assemble in the membrane (Klose, M., MacIntyre, S., Schwarz, H., and Henning, U. (1988) J. Biol. Chem. 263, 13297-13302). Linkers were inserted between the codons for residues 164 and 165 of the mutant protein. Of 13 different genes recovered, five encoded proteins which had regained the ability to assemble in the membrane. The properties of the mutant proteins, together with a structure prediction method, indicate the following rules for the final beta-strand to be compatible with, or possibly initiate, membrane insertion: (i) it must be amphiphilic or hydrophobic while its primary structure as such is fairly unimportant, (ii) it must extend over at least 9 residues, and (iii) it must not contain a proline residue around its center. One of the genes recovered coded for OmpA up to residue 164 and then followed by 10 linker-encoded residues. This 174-residue polypeptide was assembled in the membrane but did not, in contrast to all other proteins, expose sites sensitive to trypsin at the inner face of the membrane. This behavior agrees perfectly well with the OmpA model.