Production of interferon byTheileria annulata-andT. parva-infected bovine lymphoid cell lines

Theileria annulata andT. parva-infected lymphoblastoid cells were examined for their capacity to produce interferon (IFN). Supernatants of such cells were tested in biological assay for their antiviral activity. OnlyT. parva-infected cells of T-cell origin were capable of producing IFN-gamma. Supernatants of some but not allT. annulata-infected cells showed also antiviral activity, which was greatly reduced after exposure to a pH of 2. Northern-blot analysis of the cells using an IFN-gamma cDNA probe confirmed the results obtained forT. parva-infected cells in a biological assay. No IFN-gamma mRNA was detected inT. annulata-infected cells. The importance of IFN for the pathogenesis of theileriosis is discussed.     

Production of mouse mammary tumor virus upon transfection of a recombinant proviral DNA into cultured cells

We have investigated the intracellular proteins synthesized in rat XC and feline kidney cells transfected with endogenous mouse mammary tumor virus (MMTV) proviral DNA. The endogenous provirus GR40, associated with the Mtv-8 locus, directs the synthesis of gag proteins indistinguishable from those found in MMTV-infected cells. The env precursor Pr73env and the mature gp52 proteins could not be detected in these cells. Instead an env-related protein of 68K is synthesized. In contrast to this endogenous provirus, a cloned exogenous proviral variant directs the synthesis of apparently normal env proteins upon transfection into the same cell lines. These results suggest that the env gene of the endogenous MMTV provirus GR40 is defective. The exogenous proviral variant is not expected to synthesize virus particles since it carries a rearrangement in the gag gene. In order to obtain an MMTV provirus capable of correctly expressing both gag and env functions, we have constructed a hybrid endogenous-exogenous provirus containing the 5' long terminal repeat (LTR)-gag of GR40 and the pol-env-3' LTR of the exogenous provirus. Upon transfection into feline kidney cells, this hybrid provirus directed the synthesis of apparently authentic gag and env proteins. Further, virus particles can be detected in the culture medium of the transfected cells by electron microscopy. Viral proteins obtained from viral particles banded in a sucrose gradient were detected by immunoprecipitation.     

Biochemical Binding and Distribution of Protactinium-233 in the Rat

Summary

Following intravenous injection into male Sprague–Dawley rats ²³³Pa, like other elements, deposits predominantly in the skeleton (ca. 70–80 per cent), but unlike Pu and Am the liver deposition of ²³³Pa is low, about 2–3 per cent between 1 and 7 days. About 99 per cent of the injected ²³³Pa is lost from the plasma compartment in 3 days, a clearance comparable to that of Pu but much slower than that of Np, Am or Cm. On entering the liver cell cytosol ²³³Pa is bound rapidly to an unidentified protein of molecular mass 200 kDa and to a protein of 80 kDa, which is probably transferrin. Within a few hours the metal migrates to bind to a protein of > 400 kDa which has been tentatively identified as ferritin. Some ²³³Pa remains bound to small ligands until virtually all the intracellular ²³³Pa has been deposited in the lysosomes, or to a lesser extent in some other, as yet, unidentified organelles.     

The effect of mercuric chloride on the excretion of two urinary enzymes in the rat

The effect of intravenous injections of HgCl₂ on the renal excretion of alkaline phosphatase (AP) and leucine aminopeptidase (LAP) was investigated in rats. On the first day after Hg enzyme excretion showed a linear rise with the Hg dose from a threshold value of 0.44 mg Hg/kg. On the second day a statistically significant effect was seen already after 0.25 mg Hg/kg. After doses of 0.75 mg/kg or more a decrease of enzyme activity below control values occurred which persisted for more than 4 days.Treatment with 2,3-dimercaptopropansulfonate (DMPS) brought about a normalization of AP excretion. An effect on LAP excretion was observed only with early treatment. The same holds for the effect of DMPS on Hg-induced lethality.The usefulness of a measurement of LAP excretion for estimating the exposure to inorganic mercury is discussed.

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Zusammenfassung Die Wirkung von intravenös verabreichtem HgCl₂ auf die renale Ausscheidung von alkalischer Phosphatase (AP) und Leucinaminopeptidase (LAP) wurde an Ratten untersucht. Am ersten Tag nach Hg-Injektion erfolgte ein Anstieg der Enzymausscheidung, der von einem Schwellenwert von 0.44 mg Hg/kg ausgehend eine lineare Abhängigkeit von der Hg-Dosis aufwies. Am zweiten Tag wurde ein statistisch signifikanter Effekt bereits nach 0.25 mg Hg/kg beobachtet. Nach Dosen von 0.75 mg Hg/kg oder mehr fand ein Abfall der Enzymaktivität unter die Kontrollwerte statt, der mehr als 4 Tage anhielt.Behandlung mit 2,3-Dimercaptopropansulfonat (DMPS) bewirkte eine Normalisierung der AP-Ausscheidung. Eine Wirkung auf die LAP-Exkretion wurde nur bei baldiger Verabreichung von DMPS beobachtet. Das gleiche gilt für den Effekt von DMPS auf die Hg-induzierte Letalität.Die Nützlichkeit einer LAP-Bestimmung im Urin zur Abschätzung einer Hg-Inkorporation wird diskutiert.

     

Radioresistance of rat glioma cell lines cultured as multicellular spheroids. Correlation with electrical cell-to-cell-coupling

Two chemically induced rat glioblastomas, RG2 and F98, were cultured as monolayers and as multicellular spheroids and subjected to Co-gamma-irradiation. In parallel, intercellular communication between cells was determined as electrical coupling between neighbouring cells using micro-electrode techniques. A third glioblastoma with known radiobiological response (9L) was assayed with respect to intercellular communication and included into this analysis. Electrical coupling was low for RG2, intermediate for F98, and high for 9L. Radioresistance of spheroids, as expressed in terms of the mean inactivation dose computed from the survival curves increased in the same direction (RG2: 2.4 Gy; F98: 5.1 Gy; 9L: 6.5 Gy). A comparison of these parameters demonstrates a correlation between solid tumor radioresistance and gap-junctional cell-to-cell communication, at least for the class of glioblastomas analysed in this study.     

CD44 splice variants confer metastatic behavior in rats: homologous sequences are expressed in human tumor cell lines.

One of several splice variants of CD44 expressed in metastasizing cell lines of rat tumors has been shown to confer metastatic potential to the non-metastatic variant of a rat pancreatic carcinoma line (U. Günthert et al., Cell, 65: 13-24, 1991). The variant-specific rat CD44 sequences were used to detect RNA expression in human cell lines: in carcinoma lines from lung, breast and colon; and in keratinocyte lines. By polymerase chain reaction amplification, complementary DNAs encoding human homologues were isolated and sequenced. The largest splice variant has been found in a large cell lung carcinoma line and in keratinocyte cell lines. It carries at least 5 additional domains (exons) encoding a total of 338 amino acids in the membrane-proximal extracellular region of the standard CD44. Various alternative splice products have been detected in other human tumor cell lines. The distribution of CD44 splice variants is consistent with the speculation that they fulfill functions in only a few restricted differentiation pathways and that in tumor cells these pathways have been reactivated.     

Cytotoxic effects of quartz and chrysotile asbestos: in vitro interspecies comparison with alveolar macrophages

Cytotoxic effects of DQ12 quartz and chrysotile asbestos on alveolar macrophages of different animal species were compared in vitro. The type of cell reaction toward the cytotoxic dusts was always the same: a loss of cell viability (trypan blue dye exclusion test) was accompanied by the release of cytoplasmic and lysosomal enzymes. The extent of cellular destruction depended upon the amount of dust applied. In the range of 50-100 micrograms/ml quartz or chrysotile asbestos, species-specific variations were observed in the sensitivity of the cells. At this concentration alveolar macrophages of dogs, monkeys, and human patients were damaged to a greater extent than the cells from rats and cattle. Simultaneous incubation of the cells with quartz and L-alpha-dipalmitoyl lecithin resulted in a reduction of the cytotoxic quartz effect. The extent of the protective effect varied according to the species. In the case of chrysotile asbestos no reduction of the fibers cytotoxicity was observed in the presence of L-alpha-dipalmitoyl lecithin.     

Tierexperimentelle Untersuchungen zur Behandlung der akuten Eisenvergiftung

Zusammenfassung Es wurde der Einfluß von Hexacyanoferrat(II), Desferrioxamin, Diäthylentriaminpentaessigsäure und anderer Polyaminopolycarbonsäuren auf die Letalität von Mäusen nach oraler Vergiftung mit FeSO₄ untersucht. Bei frühzeitiger oraler Applikation zeigt das Hexacyanoferrat(II) eine gesichert höhere Antidotwirksamkeit als Chelatbildner. Zu späteren Zeitpunkten erwies sich die orale Verabreichung aller Antidote als unwirksam, jedoch führt die parenterale Verabreichung von Chelatbildnern zu einer deutlichen Herabsetzung der Letalität. Stärkere Unterschiede in der Wirksamkeit von Desferrioxamin, Diäthylentriaminpentaessigsäure und der anderen untersuchten Chelatbildner liegen dabei nicht vor.