m-OCTOPAMINE, p-OCTOPAMINE AND PHENYLETHANOLAMINE IN RAT BRAIN: A SENSITIVE, SPECIFIC ASSAY AND THE EFFECTS OF SOME DRUGS

Abstract— An enzyme radiochemical assay for p -octopamine, m -octopamine (norphenylephrine) and phenylethanolamine based on the N -methylation of these amines by the enzyme phenylethanolamine N -methyl transferase ( S -adenosyl- l -methionine: phenylethanolamine N -methyl transferase (EC 2.1.1.28) has been developed. [³H]Methyl- S -adenosyl- l -methionine was used as methyl donor. The reaction products are converted to their dansyl derivatives and separated by TLC in three different solvent systems prior to liquid scintillation counting. The method exhibits a sensitivity of less than 10 pg for each amine and is suitable for the measurement of endogenous p -octopamine levels in mammalian brain. The highest levels of p -octopamine were found in the hypothalamus (3.4 ng/g) but despite the sensitivity of the assay, neither phenylethanolamine nor m -octopamine could be detected. After MAO inhibition, however, both of these amines were found to be present. p -Octopamine was increased substantially in all brain regions following the administration of an MAO inhibitor, whereas pretreatment with reserpine produced a significant decrease in the hypothalamus.     

Plasmodium berghei: inhibition by splenectomy of a paralyzing syndrome in infected rats

In recent attempts to isolate the factor causing paralysis in rats infected with a mousepassed KBG 173 strain of Plasmodium berghei Splenectomy was employed. The effects of Splenectomy on the paralyzing syndrome are discussed in the present report. Paralysis was inhibited in rats splenectomized prior to inoculation. Serial bloodpassaging of the strain in the splenectomized host however, apparently enhanced its virulence. Spleen-intact rats used as controls exhibited a marked increase in incidence of paralysis. Rats splenectomized a day before paralysis became evident were paralyzed significantly more frequently than those splenectomized a day earlier, indicating the apparent requirement of an incubation period for the expression of the paralytic effect. The enhanced virulence did not appear to be related to the level of parasitemia of the splenectomized rats used as donors. The spleen appears to provide the optimal conditions required for the elaboration of the paralyzing factor.     

Effect of ethionine on dolichyl phosphate-dependent transglycosylation reactions in rat liver

Dolichyl phosphates of different chain length (C35, C55 , C75 , Dol-mixture of C90 , 95, 100, 105 and C110 ) were tested as lipid acceptors in transglycosylation reactions. In the absence of exogenously added dolichyl phosphates there were no differences in the rate of synthesis in liver of dolichyl phosphate mannose, dolichyl phosphate glucose and dolichyl pyrophosphate N-acetylglucosamine between normal and ethionine-treated animals. Addition of exogenous dolichyl phosphates of different chain length stimulated the synthesis of dolichyl phosphate mannose and dolichyl pyrophosphate N-acetyl-glucosamine 2 to 4 times depending on the chain length of dolichols , both in normal and ethionine-treated animals. In liver of ethionine-treated rats the formation of dolichyl phosphate glucose was not stimulated. Following ethionine treatment the concentration of free and esterified with fatty acids dolichols was increased.     

Degradation of adenine nucleotides in the erythrocytes of patients with chronic renal failure

In chronic renal failure AMP-deaminase operates in the erythrocytes at a much higher velocity than in healthy subjects, with a simultaneous shift from the AMP-adenine-inosine-hypoxanthine pathway (55% and 19%, respectively) to the pathway initiated by AMP-deaminase (45% and 81%, respectively).     

Independent genes coding for three acidic proteins of the large ribosomal subunit from Saccharomyces cerevisiae

The yeast ribosome contains three acidic proteins, L44, L44', and L45, closely related from a structural point of view, that seem to play a functional role similar to that of proteins L7 and L12 in the bacterial ribosome. By screening a cDNA bank in lambda gt11 with specific polyclonal and monoclonal antibodies, recombinant phages expressing each one of the acidic proteins have been cloned. A unique copy of each gene is detected using the phage cDNA inserts as probes in nitrocellulose blots of yeast DNA digested with different restriction enzymes. The inserts were subcloned in the plasmid pUC19, and their physical maps and nucleotide sequences were determined. By using the cDNA inserts as probes in genomic DNA banks, DNA fragments carrying the acidic protein genes have been cloned, characterized, and sequenced. The results conclusively show that the three yeast acidic proteins are coded by independent genes and are not the result of a post-translational modification of the product of a unique gene, as in bacteria. Like most ribosomal protein genes, the gene for protein L44' has an intron and two upstream stimulatory boxes (UASrpg) fitting closely to the consensus sequence. The genes coding for proteins L44 and L45 lack introns and seem also exceptional in other characteristics of their sequences. Proteins L44 and L45 have amino acid sequences with about 80% similarity. Protein L44' is only 63% similar to the other two polypeptides. The three proteins have highly conserved carboxyl termini comprising the last 30 amino acids, and the first 10 amino acids of L44 and L45 are identical. The results cast doubts about the possibility of a similar role for the different acidic ribosomal proteins.     

Crystallins in the differentiation and regeneration processes of the crystalline lens in amphibia

The published and authors' data have been summarized on (1) the spectrum and properties of crystallins in different amphibian species, (2) localization and synthesis of crystallins in different cellular compartments of the adult amphibian lens, (3) dynamics of crystallin formation during embryogenesis and (4) lens regeneration from tissues of the larval and adult amphibian eyes. The necessity of more detailed studies of crystallin synthesis during embryogenesis and lens regeneration using molecular biological and biochemical methods is stressed. The significance of this approach is illustrated by the pioneering data of Soviet scientists on crystallin polypeptides and corresponding mRNAs in development of Rana temporaria obtained with the use of DNA-RNA hybridization and immunoelectroblotting.     

Dynamics of adenylate cyclase desensitization during long-term exposure to the hormone

The dependence of adrenal gland adenylate cyclase desensitization on the dose of in vivo injected ACTH, the time of occurrence and duration of the enzyme refractory period and the dependence of desensitization on the number of ACTH injections were analyzed. The experiments were carried out on guinea pigs injected with prolonged action preparations of ACTH (4 and 6 units) daily for 1-6 days. Intramuscular injections of ACTH caused adenylate cyclase refraction to the repeated action of the hormone. The effect of desensitization was the most conspicuous within the first few hours after hormone injection. The decrease of adenylate cyclase sensitivity and the duration of this effect were found to depend on the ACTH dose as well as on the number of injections. It has been shown for the first time that a single in vivo injection of 0.9% NaCl causes short-term desensitization of adenylate cyclase to the repeated action of much higher doses of ACTH in vitro, presumably due to endogenous ACTH release in response to weak stress exposure. The periodicity of changes in adenylate cyclase sensitivity upon prolonged hormone administration is discussed. Sensitization of the enzyme upon daily short-term exposure to physiological doses of ACTH (administration of 0.9% NaCl for 6 days) was revealed.