5-Formyl-2'-deoxyuridine: cytostatic and antiviral properties and possible modes of action.

5-Formyl-2'-deoxyuridine (fdUrd) was prepared by a new method starting from thymidine and investigated for its influence both on proliferation of cultured mammalian cells and virus replication in vitro. The compound was found to have strong cytostatic and antiviral properties: 50% inhibition of proliferation of BHK 21/C13 cells or Ehrlich ascites tumour cells (EAT) was obtained at 4 - 10(-6) and 6 - 10(-6) M, respectively, while the treatment of pseudorabies virus with the same concentration resulted in about 1.5 log reduction of virus yield. A concentration of 1 - 10(-4) M inhibited cell proliferation by 80 to 100% while the virus yield was reduced by more than 3 orders of magnitude. All inhibitions can be prevented by thymidine.--DNA synthesis of EAT cells in vitro, as estimated by incorporation of [32P]-phosphate or low concentrations of [3H]-thymidine, was inhibited. Further biochemical experiments have provided indirect evidence that the compound is phosphorylated by thymidine and thymidylate phosphorylating enzymes. An inhibition of cell free DNA synthesis was found to be depending on a given period of preincubation with the compound (supposed to be needed for the formation of fdUrd 5'-triphosphate). This suggests that the 5'-triphosphate of fdUrd is an inhibitor of DNA polymerases and--by analogy with experiments with 5-formyluridine-5'-triphosphate and RNA polymerases [14]--may be used as an affinity label for this group of enzymes. It is concluded that the described cytostatic and antiviral effects of fdUrd are due to an intracellular "lethal" synthesis of the relevant phosphates which inhibit thymidylate synthetase (as had been found earlier to occur with the chemically prepared nucleotide in cell free extracts [1, 2]) and DNA synthesizing enzymes.     

Characterization of surface glycoproteins of mouse lymphoid cells

We have labeled exposed surface glycoproteins of mouse lymphoid cells by the galactose oxidase-tritated sodium borohydride technique. The labeled glyco-proteins were separated by polyacrylamide slab gel electrophoresis and visualized by autoradiography (fluorography). The major thymocyte surface proteins have molecular weights of 170,000 and 125,000. Thymocytes from TL antigen-positive mouse strains showed an additional band with a molecular weight of 27,000. Highly purified T lymphocytes contain two major surface glycoproteins with molecular weights of 180,000 and 125,000. Purified B lymphocytes have one major surface glycoprotein with a molecular weight of 210,000. When T lymphocytes are stimulated in vitro by concanavalin A or phytohemag-glutinin, the major proteins characteristic of T cells are relatively weakly labeled, but new components of lower molecular weights appear on the cell surface. A similar change is seen in B lymphocytes stimulated by Escherichia coli lipopolysaccharide. T lymphoblasts isolated from mixed lymphocyte cultures show a slightly different surface glycoprotein pattern. A polypeptide with a molecular weight of 57,000, which was labeled without enzymatic treatment by tritiated sodium borohydride alone, is strongly labeled in proliferating cells.     

Tumor-associated blocking factors: isolation from sera of tumor-bearing mice.

An isolation and partial purifications of tumor-associated blocking factors from the sera of tumor-bearing mice is described. Columns for affinity chromatography were prepared by coupling syngeneic tumor-immune antibodies to Sepharose 4B. Passage of serum through such immunoadsorbent columns removed all blocking activity from tumor-bearers' sera; subsequent elution of the absorbent with 3 M NaSCN recovered the activity. The blocking material was further purified on Sephadex G-200. The data provide evidence for the presence of antigen in tumor-associated blocking factors and are compatible with the hypothesis that blocking factors often consist of antigen and antibodies in the form of immune complexes.     

The effect of splenectomy, produced chemically by means of ethylpalmitate, on the formation of heteroagglutinins in the rat]

The formation of heteroagglutinins against human O-erythrocytes was pursued in Wistar rats splenectomized "chemically" by means of intravenous injections of ethyl palmitite. Contrary to some data in literature the initial inhibition in the formation of antibodies was replaced by a significant titre increase ranging from the 8th to the 14th day following immunization. The tween 20 contained in the emulsion of ethyl palmitic did not influence the formation of heteroagglutinins. The examinations show that a possible use of palmitic ethyl for immunosuppression is questionable.     

Bone marrow ribonucleic acid polymerase. Effect of testosterone on nucleotide incorporation into nuclear RNA.

The incorporation of 3H-UTP into RNA by isolated rat bone marrow nuclei is stimulated by testosterone. This effect is hormone and tissue specific. Using alpha-amanitine and different ionic strength conditions it was found that testosterone enhances preferentially RNA polymerase I activity. The sedimentation pattern of RNA isolated from bone marrow nuclei shows that the synthesis of RNA species within the 14-30 S range is mainly stimulated by the hormone.     

Changes in fibrinolysis components during short-term adaptation to high altitude]

In studying hemocoagulation in dogs under conditions of Frunze (760 m above the sea level) and Tuya-Ashu (3200 m above the sea level) it was shown that in the "emergency" phase of adaptation (the first three days) there was seen activation of fibrinolysin and profibrinolysin with depression of antifibrinolysins and inhibitors of profibrinolysin activators. The concentration of plasma fibrinogen at that period decreased by 100 mg%, which could promote an increase in the vascular permeability and improvement of oxygen approach to the tissues. Later, along with elevation of fibrinolysin and profibrinolysin activators there was a marked increase in the level of fibrinolysis inhibitors. Correlation of all the fibrinolysis components was established at a new level.     

Correlations between human portal and peripheral venous insulin and proinsulin concentrations under glucose infusion]

In 8 female patients carbohydrate tolerance was proved by means of glucose infusion test 3 days after cholecystectomy. Parameters analyzed in portal and peripheral vein blood are compared with that of 47 healthy persons. All patients demonstrate a pathological carbohydrate tolerance after cholecystectomy, further characterized by an increased lipolysis, a paradoxical rise of HGH, a diminished insulin secretion during the early and increased IRI output in the second phase. There is a significant positive correlation between portal and peripheral vein IRI concentration despite the rising portalperipheral venous IRI difference with raised portal venous IRI concentration. Corresponding differences for proinsulin concentrations can be established in the early phase only. Relations existing between blood glucose and IRI are shown by multiple regression analysis. They suggest that the altitude of IRI concentration is determined by previous blood glucose concentration.     

The mechanism of liver splanchnomegaly produced by a transplantable pituitary mammotropic tumor in rats.

The mechanism of liver splanchnomegaly developed in rats bearing a transplantable pituitary mammotropic tumor which secretes large amounts of ACTH and prolactin has been studied. The results indicate that in the first phase of tumor growth liver enlargement could be due to hypertrophy, and later mainly to hyperplasia which overcomes hypertrophy. The adrenal glands were found to play an essential role in the process of liver splanchnomegaly because adrenalectomy prevented the disproportional growth of the liver. Evidence is presented showing that glucocorticoids are the dominant hormones responsible for the development of liver splanchnomegaly.     

Electronmicroscopy studies on the exocrine pancreas of Wistar-Rats following treatment with Streptozotocin.

In Wistar rats the intraveneous injection of streptozotocin (65 mg/kg body weight) caused a permanent hyperglycemia. After 5 days there were lesions in the exocrine parenchyme of the pancreas and its nerve fibers. Pathological changes were found in cytoplasm, cell membrane, nucleus and all other cell organelles, too. The zymogen granules remaining after extensive degranulation may disintegrate in two different ways: 1. Shrinking of the granules and formation of a hale between granule membrane and core, the electronic density of which is decreased; indistinct demonstrability of the granule membrane and finally its decomposition. 2. Shrinking of the granules, decrease of the electron density and either homogeneous or mainly peripheral arrangement of the disintegrated material of the granules; irregular shape of the granules and splitting of their membranes.