Preparation of a crystalline tris-(N-methylpyridinium)-derivative of bovine insulin

The reaction between 2-chloro-N-methylpyridinium iodide and bovine insulin at pH 8.5 afforded a trisubstituted derivative as main product. Following purification on DEAE-Sephadex A-25, this derivative crystallized from a citrate buffer in the form of large rhombohedra very similar to native insulin. This observation was confirmed by zero-layer X-ray precession photography. Amino acid analysis indicated substitution at the B1-terminus and on two of the four tyrosine residues. In the mouse convulsion assay the derivative showed no biological activity.     

Stimulation of glycogenolysis by beta adrenergic agonists in skeletal muscle of mice with the phosphorylase kinase deficiency mutation (I strain)

The mechanism by which beta adrenergic agonist stimulate glycogenolysis in intact skeletal muscle was investigated in mice with the phosphorylase kinase deficiency mutation (I strain). Although extracts of I strain diaphragm muscle had only 3.7% of the phosphorylase kinase activity found in extracts of the control strain (C57BL), incubation of I strain hemidiaphragms in Krebs-Ringer bicarbonate buffer with either isoproterenol or epinephrine resulted in a stimulation of the rate of glycogenolysis. In C57BL diaphragms, the EC50 values for isoproterenol and epinephrine were 2 and 14 nM, respectively. With I strain diaphragms, dl-isoproterenol or l-epinephrine stimulated glycogenolysis as a linear function of the log of the drug concentration with no apparent plateau of response up to concentrations of 30 to 40 mugM. For each 10-fold increase in drug concentration, isoproterenol and epinephrine stimulated glycogenolysis in I strain muscles an additional 0.37 to 0.42 mg/g/hr, a slope in the concentration-response relationship of 0.17 and 0.37, respectively, of that measured in C57BL diaphragms at concentrations around the EC50. The highest glycogenolytic response measured in I strain hemidiaphragms (at 40 mugM isoproterenol) was 80% of the maximal catecholamine-stimulated glycogenolysis in C57BL diaphragms. Both 4 nM and 4 mugM isoproterenol, in a concentration-dependent manner, stimulated phosphorylase b to a conversion in I and C57BL diaphragms and increased cyclic adenosine 3':5'-monophosphate (cyclic AMP) concentrations. The glycogenolytic response to 10.1 nM dl-isoproterenol in both I and C57BL diaphragms was blocked by 34 nM l-propranolol but not by 34 nM d-propranolol. The response to 4 mugM isoproterenol was enhanced by the cyclic nucleotide phosphodiesterase inhibitors papaverine (27 mugM) or dl-4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (Ro 20-1724, 3 mugM). From the results of these studies, we conclude: 1) Catecholamines stimulate glycogenolysis in skeletal muscle of I mice, as in C57BL mice, by interacting with the beta adrenergic receptor, thereby increasing tissue cyclic AMP concentrations and stimulating phosphorylase b to a conversion. 2) alternative hypotheses for the mechanism of the catecholamine-stimulated decrease in glycogen concentration in I skeletal muscle-inhibition of glycogen synthesis, hyposia and 5'-AMP stimulation of phosphorylase b activity-have been ruled out. 3) the activity of the mutant phosphorylase kinase, although it is only 3.7% of that in extracts of C57BL muscle, is sufficient to produce phosphorylase b to a conversion and thereby account for the glycogenolytic response of I strain muscle to catecholamines.     

Derepressed alloantigen in transplacentally induced lung tumor coded for by H-2 linked gene

A transplacentally induced lung tumor of strain C3Hf mice grows progressively when transplanted to (C3Hf X A)F1 hybrid mice but not when transplanted to C3Hf recipients. Progressive tumor growth occurs in [(C3Hf X A)F1 X C3Hf] backcross mice inheriting the H-2a haplotype from the F1 parent. Furthermore, radioresistant immunity to the lung tumor can be induced in C3Hf mice by immunization with normal tissue of B10.A and B10.A(2R) but not of B10 or B10.A(5R) strain mice.     

Effects of hypertension on the static mechanical properties and chemical composition of the rat aorta

The contributions of the relative radius, relative wall thickness, incremental strain, incremental elastic modulus, and medial scleroprotein content to the static elastic properties of the rat aortic wall have been examined in three groups of rats. Controls, rats made hypertensive at four weeks of age, and rats whose blood pressure was lowered after 6 weeks hypertension, were studied. The results show evidence of adaptive changes in the aorta of hypertensive animals, and that irreversible alterations in the mechanical properties of the wall may be induced by a brief period of hypertension. A direct relationship between aortic medial scleroprotein content and the elastic properties of the wall is demonstrated.     

Improved microbiological procedure for determining bacitracin in premixes and mixed feeds

An improved microbiological procedure is presented for determining bacitracin in premixes and mixed feeds. Premixes are extracted with acidic 50% aqueous methanol, and mixed feeds are extracted with 50% dimethylformamide after washing with acetone to remove fats and pigments. Extracts are diluted with pH 6.0 phosphate buffer and assayed by using a 2-point assay system with Micrococcus flavus as the test organism. The simpler extraction coupled with the 2-point assay system measured bacitracin levels as low as 10 g/ton. Accuracy and repeatability of the 2-point system were at least equivalent to those for official AOAC methods.     

The use of the liver-spleen-scan for the diagnosis of congenital Intrahepatic cysts of the bile duct (M. Caroli) (authors transl)

After the short review of the pathogenesis, diagnosis and therapy for congenital intrahepatic cysts of the bile-duct (morbus Caroli), reference was made to the diagnostic significance of liver scintigraphy on the basis of two casuistic reports. Liver scintigraphy permits an exact localization of intrahepatic cysts as well as information regarding the course of the hepatopathy connected with morbus Caroli. In addition, 131-J-bromsulfon scintigraphy of the liver is a simple method for controlling the patency of the bile ducts. Our observations tend to indicate that preoperative and postoperative scintigraphic controls of the liver are practical for this disease.     

Characteristic Anthocyanin Pattern from Onions and other Allium spp

Three red onion (Allium cepa) cultivars, top onion (A. cepa var. vivi-parum), A. altaicum and chive (A. schoenoprasum) contained several or all of the following anthocyanins: 3-(6′-malonyl-3′-glucosylglucoside), 3-(3′,6′-dimalonylglucoside), 3-(6′-malonylglucoside), 3-(3′-malonyl-glucoside), 3-(3′-glucosylglucoside) and 3-glucoside of cyanidin. Trace amounts of two pelargonidin derivatives and the 3,5-diglucosides of cyanidin and peonidin were for the first time reported in red onion. Carbon NMR data showed that the sugars of the 3-(6′-malonyl-3′-glu-cosylglucoside), 3-(6′-malonylglucoside) and 3-glucoside of cyanidin were pyranoses. Substitution either by sugar or acid in the sugar 3-position of an anthocyanin has never been reported outside the genus Allium.