Effects of formamidoxime on macromolecular synthesis in regenerating liver and hepatomas

Formamidoxime caused an inhibition of [³H]thymidine incorporation into DNA in regenerating liver and transplanted hepatomas of different growth rates when administered by i.p. injection to rats. A dose level of formamidoxime (500 mg/kg body weight) which caused at least a 75% inhibition of DNA synthesis in these tissues had little or no effect on the incorporation of [³H]orotate into total RNA. After administration of formamidoxime there was no significant effect on amino acid nitrogen concentration in the tissues. The incorporation of ³H-labeled amino acids into acid-soluble material, cytoplasmic proteins and acid-insoluble nuclear proteins were either unaffected or showed only small changes after treatment of rats with the drug. In regenerating rat liver and Morris hepatomas 7787 and 7777, formamidoxime caused an inhibition of incorporation of ³H-labeled amino acids into both lysine-rich and arginine-rich histones. In the host livers of rats bearing the transplanted hepatomas, histone synthesis was less affected. The data indicated that formamidoxime causes inhibitory effects which are similar in nature and extent to those previously shown for the structurally related compound, hydroxyurea, in the regenerating rat liver and demonstrated that these effects can also be observed in liver tumors.     

Extracellular action potentials recorded from the interior of the giant esophageal cell of Ascaris

Exploration of the cytoplasm of the giant esophageal cell of Ascaris with a recording microelectrode shows the existence of shallow spaces where the microelectrode tip becomes extracellular in spite of being in the interior of the cell. When the microelectrode penetrates into these spaces from the cytoplasm, the resting potential shifts to a different level or entirely disappears. At the same time the large intracellular spikes are replaced by small transients similar to extracellularly recorded action potentials. It is concluded that such spaces are in communication with the external solution, and separated from the cytoplasm by an electrically active membrane; i.e., able to generate action potentials. Measurement of the potential differences between the interior of the spaces and the external solution shows that although some are not polarized, many spaces have a resting potential of the same polarity as that of the cytoplasm. It is suggested that although they are of larger size these spaces may be equivalent to the tubular systems which in other muscle cells are known to be involved in the spread of excitation into the cytoplasm.     

Purification and properties of coproporphyrinogenase

1. Coproporphyrinogenase has been prepared from rat-liver mitochondria and its properties have been studied. The isoelectric point was found to be around pH5.0 and the molecular weight to be 80000+/-8000. The pH optimum of the enzymic reaction was 7.4 and the apparent K(m) was of the order 0.03mm. The enzyme was destroyed by boiling and irreversible inactivation occurred below pH3.5. It could be stored at -10 degrees without loss of activity. The enzyme acts specifically on coproporphyrinogen III and does not form protoporphyrinogen from trans-2,4-diacrylicdeuteroporphyrin or its porphyrinogen. It was unaffected by prolonged dialysis and no cofactor requirement could be demonstrated. 2. Column chromatography of a partially purified enzyme preparation on Sephadex G-200 was found to be an improved method of purification, which gave a coproporphyrinogenase 58-fold purified. The purified enzyme was studied electrophoretically but no evidence was obtained to suggest that more than one enzyme was involved in the reaction. 3. The action was studied of various compounds added to the system. The presence of monothiol groups in the enzyme system was indicated, whereas vicinal dithiol groups were not involved in the reaction. Metal-chelating agents did not inhibit the reaction and no requirement for the presence of any essential metal has been found. All attempts to demonstrate the presence of a prosthetic group, in particular flavines, failed. Neither pyridoxal phosphate nor ATP was involved in the reaction, nor was a mitochondrial electron-transport chain required for the activity of the enzyme. Some circumstantial evidence was obtained to suggest that cis-2,4-diacrylicdeuteroporphyrin is an intermediate in the reaction.     

Acetylcholine sensitivity of the spine-test articular capsule of the sea urchin Eucidaris tribuloides

1. The changes in the consistence of the spine-test articular capsule, or ligament, of the primary spines of Eucidaris tribuloides induced by acetylcholine (ACh) have been studied. Two complementary techniques were used: (a) "forced-vibration", which detects variations in the stiffness of the ligament along a single diametral plane; and (b) "forced-rotation" which records the spatial distribution of those changes. 2. ACh (1 microM to 1 mM) caused a rapid increase in the resistive force opposed by the ligament to passive stretching. Similar effects were elicited by several monoquaternary, N-substituted derivatives of trimethylammonium. 3. The opposite effect, i.e. softening, was induced by decamethonium, dimethylphenylpiperazine, and 2-ketoamyltrimethylammonium. 4. The involvement in these effects of ACh-binding groups with pharmacological properties similar to those of the "anionic sites" of nicotinic ACh receptors is suggested.     

Detection of Petunia Asteroid Mosaic, Carnation Ringspot and Tobacco Necrosis Viruses in Ditches and Drainage Canals in a Grapevine-Growing Area in West Germany

A number of virus isolates were obtained from ditches and drainage canals in the Palatinate grapevine-growing area (‘Rheinpfalz’) in West Germany. By means of the immunoelectron microscopical decoration and the agar gel double diffusion tests these isolates were identified as petunia asteroid mosaic virus, carnation ringspot virus and tobacco necrosis virus strain D, respectively.     

Quantitative phenotypical expression of three mutant genes in barley and the basis for defining an ideotype for Mediterranean environments

Summary Three mutants induced in the two-rowed barley variety Beka and their three binary recombinants have been used in an attempt to define an ideotype suitable for Mediterranean agroclimatic conditions. Physiological methods (classical plant growth analysis) together with the study of genotype x environment interaction for grain yield were used to characterize the genotypes. That characterization brought out the huge phenotypical variation produced by only three mutant genes, suggesting that single Mendelian genes may alone explain the quantitative variation, including grain yield, without the necessity of using the polygenic concept. The genotype best adapted to the environments studied is later in heading and has shorter straw and denser spikes than Beka; it also has higher inverse of leaf area rate and grain: leaf area ratio, a lower rate of leaf senescence, and a shorter grain filling period than the original variety.     

Chronic instrumentation and longterm investigation in the fetal and maternal baboon: tether system, conditioning procedures and surgical techniques

A tether system, conditioning procedures and surgical techniques were designed to maintain chronic catheters and electrodes in the pregnant baboon and her fetus. The tether system was comprised of a lightweight metal backpack containing catheters and electrodes, couplers, pressure transducers and electrical cabling. The backpack was held snugly in place by shoulder and body straps. A flexible metal tether connected the pack to a ball bearing assembly mounted on the top of the animal's home cage. Attached to the assembly were two infusion pumps, fluid reservoir and slip ring electrical connector. The entire system rotated freely with the movements of the animal; thus, the instrumentation and connectors were secure while access was maintained for continuous physiologic recording and intravascular infusion or intermittent blood sampling with minimal physical restraint. Animals were conditioned to accept the system prior to pregnancy and animals who demonstrated tolerance were bred. An initial group of 10 pregnant animals were sham tethered during pregnancy at 102 +/- 7 days with term gestation estimated at 180 days. Surgical procedures were done at 136 +/- 4 days with placement of catheters in the maternal femoral artery and vein, fetal carotid artery jugular vein and trachea, amniotic fluid cavity, and electrodes for fetal electrocardiogram and electroencephalogram. The mean fetal survival time was 9.3 (range 0 to 29) days. The major complications which led to early delivery were placental abruption and rupture of amniotic membranes. With ultrasonic localization of the placenta and determination of fetal position before surgery, these complications may be avoided.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of linoleate,palmitate and acetate metabolism in rat ventral prostate

Rat ventral prostate incorporated (1-¹⁴C)acetate, (1-¹⁴C)palmitate and (1-¹⁴C)linoleate into different phospholipids in a time-dependent process. The rate of incorporation into total phospholipids was higher with linoleate (10.0 nmol/g) than with either palmitate (5.8 nmol/g) or acetate (4.7 nmol/g). Predominant labelling with all the radioactive substrates assayed was found in choline glycerophospholipids (PC). The radioactive profiles for linoleate in the other ventral prostate phospholipids differed from those obtained with palmitate and acetate. Specifically linoleate was incorporated into inositol glycerophospholipids plus lysoethanolamine glycerophospholipids (PI+LPE) and not into sphingomyelin (SM), while palmitate and acetate incorporated into SM but not into PI+LPE. Acetate showed the highest oxidation to CO₂ whereas no differences were observed in the radioactivity incorporated into CO₂ from a saturated (palmitate) or an essential unsaturated fatty acid (linoleate). These studies also show zinc-dependence by the acetate to CO₂ oxidation.Abbreviations PL total phospholipids - PC choline glycerophospholipids - PE ethanolamine glycerophospholipids - PI+LPE inositol glycerophospholipids plus lysoethanolamine glycerophospholipids - PS serine glycerophospholipids - SM sphingomyelin