Ibaraki Virus,an Agent of Epizootic Disease of Cattle Resembling Bluetongue

Replication of Ibaraki virus was not inhibited by 5-iodo-2′-deoxyuridine, indicating that the virus is an RNA virus. The virus was resistant to ether, chloroform and deoxycholate, sensitive to trypsin, very labile at acidic pH but stable at pH 6.4 or higher, and was resistant to repeated freezing and thawing. The virus was readily inactivated at 56 C or higher, was fairly stable at 37 C, and very stable at 4 C, while it rapidly lost infectivity when stored frozen at —20 C. The virus was readily sedimented by centrifugation at 40 000Xg for 60 min. It readily passed through membrane filters of 200 mμ pore size, passed through 100 μfilters but only with some titer loss and did not through 50 mμ filters. In these tests, the bluetongue virus used as a control behaved in the same manner as Ibaraki virus. These findings provide additional evidence for the similarity of Ibaraki virus to bluetongue virus which had been previously demonstrated on the basis of seasonal incidence, symptomatology and pathology of the diseases caused by these viruses and the behavior of the viruses in cell cultures, embryonated eggs and laboratory animals. The present study, however, provided no evidence for any serological relation between these two viruses. More Information is needed to reach a final decision on the classification of Ibaraki virus, particularly regarding the morphology of the virion, the doublestrandedness of the viral RNA and other basic features.     

Bovine Diarrhea Virus

Five strains of bovine diarrhea virus were isolated from Japanese cattle using bovine tissue cultures. These are the first isolations of this virus from Japanese cattle to be reported. Of importance is the finding that the new isolates, which are non-cytopathogenic, induce an exaltation of Newcastle disease virus in bovine testicular cell culture. This finding has provided a laboratory tool whereby the assay of the virus and its neutralizing antibody can readily be performed.     

Separation of Cl-Containing Chlorophylls by Column Chromatography

Using Toyopearl and cyclohexane: cyclohexanol solvent, fourCl-containing Chls were separated from ³⁶Cl-labeled cells ofthe blue-green, Plectonema boryanum. In normally grown cells,all four Cl-containing chlorophylls amounted to less than 1/2,000of the total Chi and about 1/50 of P700, values much lower thanpreviously reportedcontents of Chi RC I, and varied from algato alga. The level of Cl-containing Chi was markedly enhancedwhen the cells were poisoned with methyl viologen. These resultssuggests that these Cl-containing Chls are not related to thereaction center of PS I. (Received June 23, 1987; Accepted September 17, 1987)     

Purification and properties of haloalkane dehalogenase from Corynebacterium sp. strain m15-3

A haloalkane dehalogenase was purified to electrophoretic homogeneity from cell extracts of a 1-chlorobutane-utilizing strain, m15-3, which was identified as a Corynebacterium sp. The enzyme hydrolyzed C2 to C12 mono- and dihalogenated alkanes, some haloalcohols, and haloacids. The Km value of the enzyme for 1-chlorobutane was 0.18 mM. Its molecular weight was estimated to be 36,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 33,000 by gel filtration. The isoelectric point was pH 4.5. The optimum pH for enzyme activity was found to be 9.4, and the optimum temperature was 30 to 35 degrees C. The enzyme was stable for 1 h at temperatures ranging from 4 to 30 degrees C but was progressively less stable at 40 and 50 degrees C.     

Quantitative immunogold localization of Na, K-ATPase along rat nephron.

Ultrastructural localization of Na, K-ATPase alpha-subunit along rat nephron segments was investigated quantitatively by immunogold electron microscopy on LR-White ultrathin sections using affinity-purified antibody against alpha-subunit of the enzyme. Ultrathin sections were incubated with the antibody at a saturation level and the number of gold particles bound per micron of the plasma membrane (particle density) of the tubular epithelial cells from the proximal tubule to the collecting duct was determined. In all the tubular epithelial cells, gold particles were located exclusively on the basolateral surface, and no significant binding of gold particles to the apical surface was observed. Distribution of gold particles on the basolateral membranes was quite heterogeneous; lateral membranes and infolded basal membranes were highly labeled, whereas the basal membranes which are in direct contact with the basal lamina were scarcely labeled. The average particle density on the basal surface was highest in the distal straight tubule cells (11.4 units), very high in the distal convoluted tubule cells (9.8 units), intermediate in the proximal tubule cells (3.3 units), in the connecting tubule cells (4.3 units), and in the principal cells of the collecting duct (5.6-3.8 units), low in the thin limb of Henle's loop (1.0 unit), and at the control level in the intercalated cells in the connecting and collecting duct. The relative number of gold particles/mm nephron segment and the relative number of gold particles in the various nephron segments were calculated using quantitative morphological data. The estimated distribution profile of the former was in good agreement with the Na, K-ATPase activity profile in rat nephron, which was determined biochemically with a microenzymatic method.     

The combined effect of lymphokine activated killer cell and radiation therapy on rat brain tumorin vitro

Thein vitro effect of a combined treatment with lymphokine activated killer (LAK) cell and radiation therapy on rat brain tumor was examined using⁵¹Cr release assay. The tumor cell-line used in this experiment was 9L rat brain tumor derived from a Fischer 344 rat. LAK cells were obtained by culturing rat lymphocytes with recombinant human interleukin 2 for at least 3 days. The cytotoxic activity of the LAK cells was examined by⁵¹Cr release assay. Irradiation was done by exposing the microtiter plate in which the¹⁵Cr labeled 9L cells and LAK cells were cultured to a¹³⁷Cs gamma cell unit. Without irradiation, there was 18% cytotoxicity in the 1:100 tumor-to-LAK cell ratio specimen after 24 hrs cocultivation. However, if 5 Gy of irradiation was given, followed by 12 hrs incubation, the cytotoxicity was enhanced significantly at the same cell ratio (30%). This enhancement effect was the most prominent when the cell ratio was 1:100 and the irradiation dose was 5 Gy. To generate the enhancement effect, an incubation time of over 8 hrs both before and after irradiation was required. The supernatant of the LAK cells showed 19.8% and 11.4% cytotoxicity with and without irradiation, respectively. This result indicates the participation of a cytotoxic factor released from LAK cells.This work is supported in part by grant from Univeristy of Tsukuba Project Research.     

Immigration of the brown planthopper, Nilaparvata lugens, exhibiting various responses to density in relation to wing morphism

Considerable inherent variations in the relation between macropterous and brachypterous wing forms, and nymphal density were found in field populations of the brown planthopper, Nilaparvata lugens Stål (Homoptera: Delphacidae), collected from various locations in Japan. When compared under uniform laboratory rearing conditions, most of the female populations exhibited higher ratios of macropters with increasing nymphal density, but some showed extremely high proportion of brachypters and the others were highly macropterous, over broad ranges of density. These results indicate the possibility that the planthoppers in Japan, which are known not to persist in winter, are derived from different migration sources.About ten generations of successive selection for brachyptery from a population showing usual density-dependent wing morphism generated populations similar to highly brachypterous ones mentioned above. Genetic analysis of the inheritance of wing morphism revealed that brachyptery in the females was controlled by a single pair of dominant alleles. However, in the males wing forms did not segregate so clearly in the crossing experiments. This suggests that wing morphism in N. lugens in under sex-limited inheritance.

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Einwanderung von Nilaparvata lugens mit unterscheidlicher Reaktion auf Populationsdischte bei der Flügelausbildung

Zusammenfassung In Feldpopulationen von Nilaparvata lugens Stål., welche in verschiedenen Regionen Japans gesammelt wurden, bestand in der Beziehung zwischen makropteren bzw. brachypteren Flügelformen und der Larvendichte eine beträchtliche Variation. Unter einheitlichen Zuchtbedingungen im Laboratorium stieg der Makropterenanteil bei den meisten Weibchenpopulationen mit steigender Temperatur; bei einigen Populationen hingegen war entweder der Brachypterenanteil oder der Makropterenanteil extrem hoch und zwar über weite Dichtebereiche. Dies deutet auf die Möglichkeit hin, dass die Zikade in Japan, wo sie bekanntlich nicht überwintert, jeweils aus verschiedenen Quellen einwandert.Wenn eine Population mit der üblichen dichteabhängigen Flügelausbildung 10 Generationen lang auf Brachypterie selektioniert wurde, entstanden Populationen, die den erwähnten hochbrachypteren Populationen aus dem Feld glichen. Die genetische Analyse der Vererbung der Brachypterie ergab, dass bei Weibchen ein einzelnes dominantes Allel verantwortlich ist. Bei Männchen dagegen trennten sich bei Kreuzungsexperimenten die Flügelformen nicht so klar. Dies deuted auf Unterschiede zwischen den Geschlechtern bei der Vererbung der Flügelformen.

     

Level of translatable messenger RNA coding for argininosuccinate synthetase in the liver of the patients with quantitative-type citrullinemia

Summary The translation activity of mRNA coding for argininosuccinate synthetase in total RNA extracted from the liver of three patients with quantitative-type citrullinemia was determined using a cell-free translation system. In two patients, the hepatic content of the enzyme was about 20% of the control value, whereas translatable mRNA level for the enzyme was similar to or slightly lower than those of control livers. In the third patient, the enzyme content was about 50% of the control value, and mRNA activity for the enzyme was low normal. These results indicate that at least in the first two patients, the decrease in the enzyme protein is due either to increased degradation of the enzyme or to decreased translation in the patient's liver.