A rapid method for the purification of supercoiled PM2 DNA by affinity chromatography on H1 histone covalently coupled to agarose.

A simple and rapid method is described for the purification of supercoiled PM2 DNA by affinity chromatography on columns of H1 histone covalently coupled to agarose. The method does not require the use of intercalating agents or ultracentrifugation procedures. Under the conditions most appropriate for purification, elution is carried out in a single step with buffered 0.7 M NaCl after the sample has been loaded onto the column in buffered 0.2 M NaCl. The DNA eluted at the higher salt concentration consists of supercoiled closed circular DNA at greater than 90% purity independently of the ratio of supercoiled to nicked circular DNA in the input mixture.     

Tubulin polymerisation in the presence of GMP-PCP

Microtubules induced by the binding of GTP or of a non-hydrolysable analog of GTP onto the exchangeable site of tubulin appear very similar according to electron microscopy and polymerisation kinetics criteria. However, we show here that the exchangeable sites or “E” sites of the tubulin subunits remain available for nucleotide exchange inside the GMP-PCP-microtubules contrary to the “E” sites inside the GDP-microtubules. Moreover, under specific conditions, GMP-PCP induces the polymerisation of tubulin into a bidimensional, pseudocrystallin structure. Such a “crystallisation” is inhibited by GTP and GDP.     

The portal vein and its main affluents. An anatomic and morphometric study

There are reported the results of certain measurements made on 158 human cadavers of both sexes. A series of main parameters are given, such as the distance between the superior mesenteric vein and on the one hand its confluence with other vessels and on the other hand the pancreas and the horizontal part of the duodenum: the distance between the superior mesenteric vein together with the portal vein, and the inferior caval vein; the diameters of the portal trunk and the confluence angle between the roots of the portal vein. The results reflect the possibility to perform a troncular portocaval anastomosis in at least 90 per cent of the cases. The authors' attention was, however, directed towards the anatomy of the superior mesenteric vein, as this vessel is preferred by some surgeons in the achievement of the portocaval anastomosis.     

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).