Outbreaks of type C botulism in waterfowl in Japan

Four outbreaks of botulism in waterfowl were encountered over a five-year period of 1973 to 1977 in Japan. In all the outbreaks toxin was detected from all 12 sera, twenty-three of 24 gizzard contents from diseased or dead birds and one of three maggots. It was neutralized with Clostridium botulinum type C antitoxin serum, regardless of its origin. By using CO2 gas jet method, C. botulinum was isolated from four of 11 gizzards from diseased birds, five of 7 ones from dead birds, one of one maggot and one of one sludge sample, that is, eleven of 20 specimens in total. All 20 strains were identical with C. botulinum type C in biological properties. Most of the isolates showed a toxin titer ranging from 1,000 to 200,000 LD50 for mice. Four of them were identified as type C by mouse neutralization tests with antitoxin sera. The toxic suspensions of a strain 1-15 were administered orally to Chinese spot-billed ducks, which died when more than 200,000 LD50 mouse toxin was administered. Environmental conditions for occurrences of waterfowl botulism were discussed.     

Effects of 2 water-soluble contrast media, meglumine iothalamate (Conray) and meglumine iocarmate (Dimer-X), on the electric activity of identifiable giant neurons of the African giant snail (Achatina fulica Ferussac)]

The influences of two water soluble contrast media, meglumine iothalamate and meglumine iocarmate, on the neuronal excitability and on the neuronal sensitivity to putative transmitters were examined in comparison with those of sucrose using two identifiable giant neurones of Achatina fulica Férussac (the TAN and the PON). A relatively low increase of osmotic pressure of the extracellular fluid, produced by the application of contrast media, reversed the Cl- dependent inhibition caused by a putative transmitter. The same increase of this osmotic pressure, however, did not influence the Cl- independent inhibition and the excitation of the neurone examined. The hyperpolarization of neuromembrane was caused by an increase of osmotic pressure of the extracellular fluid. Its relatively high increase was necessary to make spontaneous spike discharges disappear totally. All effects of the two contrast media, observed in this study, were due to the increase of osmotic pressure of the extracellular fluid ; no specific effect of the contrast media containing the iodine on the indicators used was observed.     

Classification of the responses of a giant neuron from the snail Acatina fulica Férussac, caused by inhibitory sugstances depending on chlorine ions]

GABA, three of its derivatives (l-GABOB, d-GABOB and delta-amino valeric acid), acetycholine (Ach), dopamine (DA) and l-Phe-Tyr all inhibit an identifiable giant neurone (the TAN, tonically autoactive neurone) of Achatina fulica. These effects were examined by microdrop application in two different conditions: in physiological solution and in the absence of chloride ions. The results show that the relatively transient (rapid) inhibitions caused by GABA, by its derivatives and by Ach are dependent on chloride ions; the relatively maintained (long-lasting) inhibitions, caused by DA and l-Phe-Tyr, are independent of chloride ions.     

Redescription of Caprogammarus gurjanovae Kudrjaschov & Vassilenko, 1966 (Crustacea: Amphipoda) from Hokkaido,Japan, with notes on the taxonomic status of Caprogammarus

Caprogammarus gurjanovae Kudrjaschov & Vassilenko, 1966 (Amphipoda: Caprellidea: Caprogammaridae) was redescribed based on the materials newly collected off Kushiro and Akkeshi, Hokkaido, Japan, which represents the southernmost record of this genus. Caprogammarus and Caprella share an identical feature, that of having the head and perionite I partially fused. Thus, Caprogammarus is considered to be a member of the suborder Caprellidea.     

Synthesis of lipopolysaccharide by Escherichia coli cells recovering from sublethal heat stress.

Escherichia coli cells exposed to a sublethal heat treatment at 55 degrees C for 15s synthesized lipopolysaccharide during their recovery period after heat stress. As chloramphenicol at least partly inhibited the synthesis of lipopolysaccharide, it is suggested that its synthesis might be in part due to the lipopolysaccharide-synthesizing enzymes produced de novo. The results obtained coincided with our previous finding that the permeability barrier function was repaired by heat-stressed cells.     

Inhibitory effect of mitoxantrone on activity of protein kinase C and growth of HL60 cells.

Mitoxantrone, a new anthraquinone, showed inhibitory an effect on protein kinase C (PKC) activity. Its IC50 value was 4.4 micrograms/ml (8.5 microM), which is much lower than those of the well-known anthracyclines daunorubicin and doxorubicin, the IC50 values of which are more than 100 micrograms/ml (> 170 microM). Kinetic studies demonstrated that mitoxantrone inhibited PKC in a competitive manner with respect to histone H1, and its Ki value was 6.3 microM (Ki values of daunorubicin and doxorubicin were 0.89 and 0.15 mM, respectively), and in a non-competitive manner with respect to phosphatidylserine and ATP. Inhibition of phosphorylation by mitoxantrone was observed with various substrates including S6 peptide, myelin basic protein and its peptide substrate derived from the amino-terminal region. Their IC50 values were 0.49 microgram/ml (0.95 microM), 1.8 micrograms/ml (3.5 microM), and 0.82 microgram/ml (1.6 microM), respectively. Mitoxantrone did not markedly inhibit the activity of cyclic AMP-dependent protein kinase, casein kinase I or casein kinase II, at concentrations of less than 10 micrograms/ml. On the other hand, brief exposure (5 min) of HL60 cells to mitoxantrone caused the inhibition of cell growth with an IC50 value of 52 ng/ml (0.1 microM). In HL60 cells, most of the PKC activity (about 90%) was detected in the cytosolic fraction. When HL60 cells exposed to 10 micrograms/ml mitoxantrone for 5 min were observed with fluorescence microscopy, the fluorescence elicited from mitoxantrone was detected in the extranuclear area. These results indicated that mitoxantrone is a potent inhibitor of PKC, and this inhibition may be one of the mechanisms of antitumor activity of mitoxantrone.     

Phosphorylation of histone H2A by protein kinase C and identification of the phosphorylation site.

In regenerating rat liver, nuclear protein histone H2A was shown to be phosphorylated on its amino-terminal serine residue [Sung et al. (1971) J. Biol. Chem. 246, 1358-1364], but the protein kinase which phosphorylates this residue has not been identified. To evaluate the possibility that protein kinase C can phosphorylate this residue, calf thymus histone H2A was 32P-labeled by incubation with [gamma-32P]ATP and highly purified protein kinase C from rat brain in the presence of calcium and phospholipid. About 1 mol of 32P was incorporated per mol of histone H2A and the Km and apparent Vmax of the reaction were calculated to be 2.1 microM and 0.35 mumol/min/mg, respectively. So histone H2A seemed to be a good substrate for protein kinase C. Further, the proteolytic phosphopeptides of 32P-labeled histone H2A were isolated by means of a series of column chromatographies and analyzed for their amino acid compositions. Comparison of the data with the known primary structure of histone H2A revealed their amino acid sequence as 1Ser-Gly-Arg. These data suggest that protein kinase C may be a candidate for the protein kinase which phosphorylates the amino-terminal serine residue of histone H2A during the regeneration of rat liver.     

The 72-kDa Microtubule-Associated Protein from Porcine Brain

A microtubule-associated protein (MAP) with a molecular mass of 72-kDa that was purified from porcine brain by using its property of heat stability in a low pH buffer was characterized. Low-angle rotary shadowing revealed that the 72-kDa protein was a rodlike protein approximately 55-75 nm long. The 72-kDa protein bound to microtubules polymerized from phosphocellulose column-purified tubulin (PC-tubulin) with taxol and promoted the polymerization of PC-tubulin in the absence of taxol. Microtubules polymerized by the 72-kDa protein showed a tendency to form bundles of several microtubules. Quick-freeze, deep-etch electron microscopy revealed that the 72-kDa protein formed short crossbridges between microtubules. We performed peptide mapping to analyze the relationship of the 72-kDa protein to other heat-stable MAPs, and the results showed some resemblance of the 72-kDa protein to MAP2. Cross-reactivity with a monoclonal anti-MAP2 antibody further suggested that the 72-kDa protein and MAP2 are immunologically related. To study the relationship between the 72-kDa protein and MAP2C, a smaller molecular form of MAP2 identified in juvenile rat brain, we prepared the 72-kDa protein from rat brain by the same method as that used for porcine brain. The fact that the 72-kDa protein from juvenile rat brain was also stained with our monoclonal anti-MAP2 antibody also suggested that the 72-kDa protein is an MAP2C homologue of the porcine brain.