Pituitary-adrenocortical system in patients with Shy-Drager syndrome

We examined ACTH responses to the three main mechanisms of ACTH secretion, i.e., stress, negative feedback and circadian rhythm, in six patients with Shy-Drager syndrome and in six control subjects to determine whether or not injury to the central autonomic nervous system provokes some disturbances in ACTH secretion. The patients showed a poor cortisol response to the stress of insulin induced hypoglycemia along with a normal ACTH and urinary 17-OHCS response to metyrapone and a normal cortisol circadian rhythm. The discrepancy between the above-mentioned functional tests of ACTH secretion is rare to our knowledge. These findings suggested the existence of a glucoreceptor defect in such patients, or the possibility that the stress of insulin induced hypoglycemia stimulates ACTH secretion by way of the autonomic nervous system.     

Pyruvate:NADP+ oxidoreductase from Euglena gracilis: limited proteolysis of the enzyme with trypsin

Pyruvate:NADP+ oxidoreductase from Euglena gracilis, a homodimeric protein with a molecular weight of 309 kDa, is an iron-sulfur flavoenzyme that contains thiamin pyrophosphate (TPP). The functional structure of the enzyme was studied by a limited proteolysis experiment using trypsin. The evidence obtained shows that the enzyme consists of two functional domains, one of which contains an iron-sulfur cluster, which can be isolated as a homodimeric fragment of approximately 220 kDa by proteolysis. The other domain that contains FAD is released as a monomeric fragment of approximately 55 kDa. The pyruvate dehydrogenase reaction is still catalyzed by the large fragment when NADP+ is substituted by methyl viologen, while the small fragment retains a diaphorase-like electron-transfer activity from NADPH to MV. It is thus shown that pyruvate is oxidized in a CoA-dependent reaction to form CO2 and acetyl-CoA in the iron-sulfur domain, and that the two electrons formed are transferred to the FAD domain in which NADP+ is reduced. TPP is considered to be associated in the iron-sulfur domain. The NH2-terminal sequences of the enzyme and its proteolytic fragments reveal that the iron-sulfur domain occurs in the NH2-terminal side of the enzyme. For elucidation of the O2 instability of the enzyme, limited proteolysis was attempted in air. The tryptic fragment derived from the iron-sulfur domain, similar to the native enzyme, appears to be inactivated by direct contact with O2. In contrast, the FAD domain, when separated from the other domain, is quite stable in air, although the diaphorase activity decays when the native enzyme is exposed to O2.     

Fluorescent probe and permeability to cells of isopoly (S-carboxymethyl-L-cysteine) derivative of nucleic acid bases.

Isopoly(S-carboxymethyl-L-cysteine) derivatives of nucleic acid bases were found to form stable complex with oligo-DNA in vitro. Fluorescent probed isopoly(S-carboxymethyl cysteine) derivatives of nucleic acid bases were prepared as antisense oligomers. The transfection of the oligomer into cells was carried out by HVJ-liposome method. Fluorescence was observed from the cells treated with HVJ-liposome including fluorescent probed oligomers.     

Occurrence and characterization of fructose 6-phosphate, 2-kinase and fructose 2,6-bisphosphatase in Euglena gracilis

1. Fructose 6-phosphate, 2-kinase and fructose 2,6-bisphosphatase occurred in Euglena gracilis SM-ZK, and is located in cytosol. 2. Fructose 6-phosphate, 2-kinase and fructose 2,6-bisphosphatase were partially purified, and both enzyme activities were not separated during the partial purification. 3. The pH optimum for fructose 6-phosphate, 2-kinase activity was 7.0. The saturation curve of the enzyme activity for ATP concentration was hyperbolic, and the Km value for the substrate was 0.88 mM. On the other hand, the saturation curve of the enzyme activity for fructose 6-phosphate concentration was sigmoidal, and the K0.5 value for the substrate was 70 microM. 4. The pH optimum for fructose 2,6-bisphosphatase activity was 6.5. The saturation curve for fructose 2,6-bisphosphate concentration was sigmoidal, and the K0.5 value for the substrate was 1.29 microM. Fructose 2,6-bisphosphate showed a substrate inhibition at high concentration over 5 microM, and the enzyme activity was completely inhibited by 20 microM of fructose 2,6-bisphosphate.     

Purification and some properties of short chain-length specific trans-2-enoyl-CoA reductase in mitochondria of Euglena gracilis

Short chain-length specific trans-2-enoyl-CoA reductase (reductase I), which contributed to mitochondrial fatty acid synthesis, was purified about 200-fold from crude extract of mitochondria in Euglena gracilis. It had a molecular weight of 39,000, and consisted of two dissimilar subunits with molecular weights of 15,000 and 25,000. The enzyme utilized crotonyl-CoA as the most active substrate and showed negative cooperativity in the reaction with the substrate. NADH was the sole electron donor. Some divalent cations were inhibitory to the enzyme when incubated with the enzyme prior to the start of the reaction. The reductase apparently contained loosely bound FAD.     

Identification of Leptospira andamana isolated from the spinal fluid of a fatal case of leptospirosis in S?o Paulo, 1963.

The IAL-S. P. strain was isolated from the spinal fluid of a patient male, aged 35, black, a sewer worker with fever, myalgia, jaundice, vomiting and meningitis symptoms with a 5-day incubation period after the lower half of the body had been submerged for 2 hours in sewers when unblocking a drain. Leptospires were isolated by direct inoculation of the spinal fluid taken on the 9th day of the illness into the Fletcher's media and into guinea pigs by the intraperitoneal route. The patient gave a positive agglutination test for L. andamana with cross-reaction with L. sejroe. The strain was identified as L. andamana by the cross-agglutination-lysis test and the cross-absorption test. The IAL-S. P. strain is undoubtedly not saprophytic but parasitic and pathogenic for man and animals, however its biological properties resist to the oligodynamic action of Cu and Hg and the 8-azoguanine action as in the case of the Patoc 1 strain. I could be recommended to reconsider whether the strain belongs to L. interrogans, L. biflexa or to another group because the grounds for L. andamana being saprophytic were denied by this report.