New Plasmid-Mediated Phosphorylation of Gentamicin C in Staphylococcus aureus

A clinical isolate of Staphylococcus aureus resistant to gentamicin, kanamycin and 3′,4′-dideoxykanamycin B contained two enzymes capable of inactivating gentamicin, i.e., an aminoglycoside 2″-phosphotransferase and aminoglycoside acetyltransferase.     

Pre-activation strategy for oligodeoxyribonucleotide synthesis using triaryloxydichlorophosphoranes in the phosphotriester method.

Triaryloxydichlorophosphoranes were tested as condensing agents for oligodeoxyribonucleotide synthesis in the phosphotriester method. Tris(2,4,6-tribromophenoxy)dichlorophosphorane (BDCP) was found to be a relatively stable crystalline material which could be used as a chemical reagent. A notable feature of the BDCP-promoted condensation reaction was studied by 31P-NMR. A small amount of BDCP compared to the conventional condensing agent was effective for the generation of active nucleotide intermediates and BDCP itself was quantitatively converted into an inert material, tris(2,4,6-tribromophenyl)phosphate (2). Thus, BDCP enabled us to separate the activation step from the condensation process in the phosphotriester method. This preactivation method was applied to the solid-phase synthesis.     

Mutagenesis of human granulocyte colony stimulating factor

To define the structure-function relationship, we have made a number of mutants of human granulocyte colony-stimulating factor (hG-CSF) by in vitro mutagenesis. The results indicate that most of the mutations located in the internal and C-terminal regions of the molecule abolished the activity, whereas the mutants without N-terminal 4, 5, 7, or 11 amino acids retained the activity. N-terminal amino acids were also altered by cassette mutagenesis using a synthetic oligonucleotide mixture. Among them, KW2228, in which Thr-1, Leu-3, Gly-4, Pro-5 and Cys-17 were respectively substituted with Ala, Thr, Tyr, Arg and Ser, showed more potent granulopoietic activity than that of intact hG-CSF both in vitro and in vivo.     

Clinical studies on cell-mediated immunity in patients with renal cell carcinoma: interleukin-2 and interferon-γ production of lymphocytes

Summary The authors examined interleukin-2 (IL-2) production and interferon (IFN) production of peripheral blood mononuclear cells in 28 patients with renal cell carcinoma and 17 control subjects. The peripheral blood was obtained prior to the initiation of therapeutic procedures. The patients were divided into two groups according to tumor size, 5 cm and >5 cm. The production of IL-2 and IFN was measured by immunoradiometric assay. As a result, in the patients with tumors >5 cm, IL-2 and IFN production was impaired. However, in the patients with tumors 5 cm, IFN production was enhanced, though IL-2 production was not significantly different from that of the control subjects. There was no significant correlation between IL-2 production and IFN production.     

The mechanism of low susceptibility to stress in gastric lesions of spontaneously hypertensive rats.

The mechanism of low susceptibility to stress in gastric lesion formation in spontaneously hypertensive rats (SHR) was investigated focusing on the sympathetic and parasympathetic nervous systems. In the gastric tissues of SHR, norepinephrine (NE) and dopamine (DA) contents were higher, while acetylcholine content and choline acetyltransferase activity were lower than those of Wistar-Kyoto rats (WKY). Water-immersion restraint induced gastric lesions frequently in WKY (ulcer indices : 52 +/- 7mm2) but less frequently in SHR (ulcer indices : 3 +/- 1mm2). Although NE content decreased in both SHR and WKY as a result of water-immersion restraint, it remained higher in SHR than in WKY. ACh content decreased by the procedure in WKY but not in SHR. DA content was increased by the procedure in all gastric regions of SHR. The gastric lesions induced in SHR were aggravated by pretreatment with 6-hydroxydopamine, an agent for chemical sympathectomy, following decreases of NE and DA contents. These results indicate that the relative sympathetic hyperfunction, parasympathetic hypofunction and dopaminergic mechanism in the stomach contribute to the prevention of gastric lesion formation in SHR.     

Nalidixic acid-resistant mutations of the gyrB gene of Escherichia coli

Summary DNA fragments of 3.4 kb containing the gyrB gene were cloned from Escherichia coli KL-16 and from spontaneous nalidixic acid-resistant mutants. The mutations (nal-24 and nal-31) had been determined to be in the gyrB gene by transduction analysis. Nucleotide sequence analysis of the cloned DNA fragments revealed that nal-24 was a G to A transition at the first base of the 426th codon of the gyrB gene, resulting in an amino acid change from aspartic acid to asparagine, and nal-31 was an A to G transition at the first base of the 447th codon, resulting in an amino acid change from lysine to glutamic acid. This indicates that mutations in the gyrB gene are responsible for nalidixic acid resistance.     

Fucosylation of serum alpha-fetoprotein in patients with primary hepatocellular carcinoma

alpha-Fetoprotein specimens were prepared from the sera of four patients with hepatocellular carcinoma. The lentil lectin-reactive and lectin-nonreactive variants of this glycoprotein were also prepared from the serum of one of the four patients by affinity chromatography with immobilized lectin. The correlation between the carbohydrate structure of these compounds and their reactivity in crossed immuno-affinoelectrophoresis with lentil lectin was studied by chemical analysis and affinity chromatography of the glycopeptides with lectin columns. It was found that the lentil lectin-reactive variant contained a carbohydrate chain of the fucosylated biantennary complex type. These data together with previous findings indicate that most of the patients with hepatocellular carcinoma have an elevated serum concentration of fucosylated alpha-fetoprotein.