Differentiation of canine bone marrow stromal cells into voltage- and glutamate-responsive neuron-like cells by basic fibroblast growth factor

We investigated the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage- and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons containing recombinant human basic fibroblast growth factor (bFGF; 100 ng/ml). The viability of the bFGF-treated cells was assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial markers. Western blotting and immunocytochemical analysis for the neuronal markers were performed to evaluate the protein expression and localization. The Ca²⁺ mobilization of the cells was evaluated using the Ca²⁺ indicator Fluo3 to monitor Ca²⁺ influx. To investigate the mechanism of bFGF-induced neuronal differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide 3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore, in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase in intracellular Ca²⁺ levels. Each inhibitor significantly attenuated the bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells and may lead to the development of new cell-based treatments for neuronal diseases.     

Blackening of Diospyros genus xylem in connection with boron content

The amount of boron contained in the xylem of Japanese persimmon (Diospyros kaki Thunb.) and ebony (Diospyros ebenum Koen) was determined by inductively coupled plasma (ICP) emission spectrometry and a modified curcumin–acetic acid method. The boron content was compared between the heartwood and sapwood of ebony as well as between a blackened portion (“kurogaki”) and normal portion of Japanese persimmon. The kurogaki contained a higher level of boron than the normal portion of the same individual, although the boron content varied among individuals. Moreover, the boron content of the heartwood of ebony was much higher than that of the sapwood. These results suggest the participation of boron in the blackening of Japanese persimmon. Because both kurogaki and heartwood of ebony are durable to fungal attack, the blackening of Diospyros genus appears to be related to the formation of defensive substances in which boron seems to take part. The convenient curcumin–acetic acid method is an alternative to the ICP method with comparable accuracy.     

Fog and Precipitation Chemistry at Mt. Rokko in Kobe, April 1997-March 1998

Fog water and precipitation were collected and analyzed to study fog and precipitation chemistry. The research was carried out through one year from April 1997 to March 1998 at Mt. Rokko in Kobe. Higher fog occurrence and larger volume of fog water were observed in summer, corresponding to the trend of seasonal variation in precipitation amount. The annual mean pH value of fog water (3.80) was lower by ca. one pH unit than that of precipitation (4.74). The concentration of chemical species in fog water was ca. 7 times that in precipitation. The highest anion and cation concentrations were SO₄ ^2? and NH₄ ⁺ in fog water and Cl^? and Na⁺ in precipitation, although the Cl^?/Na⁺ equivalent ratio in both fog water and precipitation was almost the same value as that in sea water. It is considered that in the longest fog event, NH₄ ⁺ and nss-SO₄ ^2? in fog water mainly scavenged as (NH₄)₂SO₄, mainly derived from (NH₄)₂SO₄ (aerosol) in the atmosphere, NH₃ was scavenged at the growing stage, and SO₂ was also scavenged after the mature stage. NO₃ ^? in this fog event was mainly absorbed as HNO₃.     

Pseudorabies virus infection in hunting dogs in Oita,Japan: Report from a prefecture free from Aujeszky’s disease in domestic pigs

We isolated two pseudorabies virus (PRV) isolates (designated OT-1 and OT-2) from two hunting dogs exhibiting neurological manifestations after eating the flesh of wild boar hunted in Oita prefecture, Kyushu Island, Japan. The isolates corresponded to a previously reported PRV (MY-1 strain) isolated from a hunting dog in neighboring Miyazaki prefecture, and it clustered into genotype II based on the glycoprotein C sequence. Our results suggest that this common PRV strain may have been maintained in wild boars on Kyushu Island even though domestic pigs in this area have attained an Aujeszky’s disease-free status.     

Germination and appressorium formation of Pyricularia oryzae Cavara can be inhibited by reduced concentration of Blasin®Flowable with carbon dioxide microbubbles

We investigated the possibility to reduce the usage of Blasin^®Flowable (BF), a disinfectant inhibiting the germination and appressorium formation of Pyricularia oryzae Cavara conidia, by using carbon dioxide microbubbles (CO₂MB). Germination was significantly inhibited by 10 000-fold diluted BF solution containing CO₂MB generated by the decompression-type generator compared to CO₂ millibubbles (CO₂MMB) and CO₂MB generated by the gas-water circulating-type generator. Appressorium formation in the 10 000-fold diluted BF solution containing both CO₂MBs was less than that in CO₂MMB. Scanning electron microscopy showed wrinkles and dents on the surface of conidia treated with 5 000-fold diluted BF solution containing both CO₂MBs. Via transmission electron microscopy, we observed the expansion of the vacuole and the intracellular space and bloated or absent lipid granules in the conidia treated with BF solution containing both CO₂MBs. Our results show that inhibition of the conidium germination and appressorium formation of P. oryzae Cavara by 10 000-fold diluted BF solution could be achieved by using the decompression-type CO₂MB.     

Thermal tolerance of a thermally selected strain of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> and the pedigrees of its F1 and F2 generations indicated by their critical thermal maxima

A thermally selected strain of rainbow trout has been established by selective breeding since 1966 in Miyazaki, Japan. In the present study, we compared the critical thermal maxima (CTMs), the temperatures at which organisms reach a predefined sublethal endpoint and lose their equilibrium, between a thermally selected and two normal (Donaldson) strains of rainbow trout. The CTM of one normal strain from Nikko (Nikko strain) acclimated to 20 °C (29.7 °C) was significantly lower than those of the thermally selected strain (30.0 °C) and the other Donaldson strain from Aomori (29.9 °C) (P?<?0.05). The F1 generations, F1T and F1N, were produced by crossing thermally selected strain females with Nikko strain males and Nikko strain females with thermally selected strain males, respectively. No significant difference was observed in the CTM between F1T [30.1?±?0.15 °C (n?=?30)] and F1N [30.1?±?0.16 °C (n?=?30)] (P?>?0.05) for fish acclimated to 20 °C, suggesting that the F1 offspring inherited the thermal tolerance trait from one thermally selected strain parent irrespective of whether it was the male or female. F2 offspring of F1T or F1N also showed the thermal tolerance trait. The coefficients of variation for CTM were also compared among all the datasets obtained in the present study and their values for F1 hybrids were lower than those of the parental generation of the Nikko strain (P?<?0.05). In contrast, the coefficients of variation of F2s were the same as those of their parental generation. Furthermore, the thermally selected strain and Nikko strain as a reference family provide a F2 generation for segregating phenotypes, which is required for in-depth genetic analysis of the thermally selected rainbow trout strain.