Immunolocalization of <Emphasis Type="Italic">Pepper mild mottle virus</Emphasis> in developing seeds and seedlings of <Emphasis Type="Italic">Capsicum annuum</Emphasis>

The location of Pepper mild mottle virus (PMMoV) within seeds as they developed on inoculated seedlings of pepper (Capsicum annuum) was followed over time by detecting the viral coat protein using immunofluorescence microscopy. Seedlings were inoculated with PMMoV when the flower buds on the first and second branching nodes were in bloom. Fluorescence indicating the presence of PMMoV was first observed around immature seeds and placentas in the ovaries on the fourth branching node at 20 days post-anthesis (20 DPA), which corresponded to 39 days post-inoculation (39 DPI). The area with fluorescence gradually expanded from the placenta into the integument and the parenchyma, and finally reached the tip of the immature seeds by 34 DPA (53 DPI). The embryo or endosperm beyond the endothelium never fluoresced during the experiment [i.e., ending at 81 DPA (102 DPI)]. For visualizing viral routes of invasion from seeds into new seedlings, PMMoV-infected C. annuum seeds that were heterozygous for the L ³ tobamovirus-resistance gene were sown in soil at 30°C. After ~2 weeks, the cotyledon developed virally induced necrosis. These findings shed light on the infection cycle of PMMoV through vertical transmission in C. annuum.     

Comparison of Iron Availability in Leaves of Barley and Rice

Iron (Fe) is an essential trace element in all eukaryotes. In higher plants, Fe deficiency causes interveinal chlorosis in young leaves. However, in barley and rice, both of which are "Strategy II" plants, the degree and the pattern of Fe-deficiency symptoms differ. In the present study, barley and rice plants were grown in the same container, i.e., by "coculturing," to compensate for the amount of mugineic acids in rice in the nutrient solution. We examined the differential availability of Fe for distribution and retranslocation in shoots between barley and rice without considering the difference in the iron acquisition ability, which is affected by the differential mugineic acid secretion between barley and rice. Although the Fe concentration of young barley leaves had decreased under the coculture conditions, the SPAD value was similar to that in monocultured barley. In contrast, although there was an increase in the Fe concentration of the young leaves of cocultured rice, the SPAD value decreased, as in the case of monocultured rice. Rice accumulated Fe in old leaves, whereas in barley Fe was efficiently distributed to young leaves. Therefore, the SPAD value of the second leaf in rice remained constantly high. The Fe concentration of the second leaf in barley decreased under Fe-deficient coculture conditions, the SPAD value decreased and the senescence of the second leaf become accelerated. ⁵⁹Fe pulse-labeling experiments suggested that in barley Fe was more efficiently retranslocated from old leaves to young leaves than that in rice. As a result, the level of Fe present in the fraction with a molecular weight lower than the 10,000/water-soluble Fe ratio was higher in the old leaves of barley than in the old leaves of rice under Fe-deficient conditions. Based on the results obtained, we suggest that the distribution and retranslocation characteristics of internal Fe in barley may be well adapted to Fe deficiency.     

Kinetics of bacterial death by heated dolomite powder slurry

The kinetics of the bactericidal action of dolomite powders heated at 600-1000 °C against Escherichia coli and Staphylococcus aureus were investigated. Dolomite powder heated to at least 700 °C exhibited bactericidal action, and the process of bacterial death in the heated dolomite powder slurries followed first-order reaction kinetics. The value of the death rate constant (k) increased with dolomite powder concentration, and the dilution coefficient (n), which indicates the dependence of k on the reagent concentration, was measured. The n values of the powder heated at 700 °C and at temperatures >900 °C were almost identical to those of MgO and CaO, respectively. This suggests that the first emergence of bactericidal action at 700 °C corresponds to generation of MgO while that at temperatures >900 °C is due to generation of CaO. The slurry temperature significantly affects the bactericidal action. The slope of the Arrhenius plot of k for E. coli and S. aureus grown at 37 °C exhibited a discontinuous point at approximately 22 °C, where a change in the value of activation energy for bacterial death occurred. This temperature corresponds to that of the phase transition of cell membrane lipids.     

Effect of cellooligosaccharide or synbiotic feeding on growth performance,fecal condition and hormone concentrations in Holstein calves

We investigated the effect of cellooligosaccharide (CE) or a combination of dextran and Lactobacillus casei ssp. casei strain JCM1134^T (synbiotic; SB) feeding on growth performance, fecal condition and hormone concentrations in Holstein calves. Fifty‐two female Holstein calves were randomly assigned to three treatment groups: CE feeding group (n = 16), SB feeding group (n = 18), and control group (n = 18). Body weight at 90 days of age, as well as daily body weight gain (DG) and feed efficiency after weaning to 90 days of age were greater (P < 0.05) in the CE feeding group than in the control group. The total fecal score tended to be lower (P < 0.1) in the SB feeding group than in the control group. Plasma insulin concentration was higher (P < 0.05) in the CE feeding group than in the control group at 90 days of age. Our results indicate that CE feeding improved DG and feed efficiency in calves. On the other hand, there was less effect on growth performance and fecal Escherichia coli counts in calves fed SB.     

Regulation of secondary follicle growth by theca cells and insulin-like growth factor 1

Ovaries contain follicles at various stages of development, including primordial, primary, secondary, antral and Graafian follicles. Although the growth of these follicles is controlled to maintain regular ovulation, the mechanism through which this occurs remains unclear. In our study, we found that the growth rate of cultured secondary follicles separated from mice ovaries differed between follicles. After 4 days of culture, the size of some secondary follicles was markedly increased, while that of others had either slightly increased, remained unchanged or shrunk. We compared the expression levels of growth factors between these secondary follicles and found that the growth rate of cultured secondary follicles correlated with the expression level of insulin-like growth factor 1 (Igf1) mRNA. Igf1 mRNA expression level in secondary follicles containing theca cells was higher than that in secondary follicles without theca cells, and the granulosa cell proliferation around follicles containing theca cells was increased. Furthermore, an IGF1 inhibitor also inhibited the granulosa cell proliferation, and administration of IGF1 to secondary follicles without growth promoted granulosa cell proliferation. These results indicated that the theca cells of secondary follicles induced the expression of IGF1 and promoted the follicle growth.     

The formation process of button ulcers in pigs experimentally infected with a subgenotype 2.1 isolate of classical swine fever virus

We evaluated the role of classical swine fever virus (CSFV) in the formation of button ulcers in the mucosa of the gastrointestinal tract. Histopathological and immunohistochemical analyses of pigs experimentally infected with a subgenotype 2.1 isolate of CSFV, which was isolated in Japan in 2019, revealed follicular necrosis in the submucosal mucosa-associated lymphoid tissue and herniation of crypts as factors that contribute to the development of button ulcers during CSFV infection. These findings indicate that CSFV induces follicular necrosis and is one of the causative agents of button ulcers in pigs.     

Large-scale survey of frequency of forest walking and related factors in a Japanese population inhabiting a large city, and comparison of an urban area and a rural area

The frequency of forest walking among the general population is a major topic of study in forest science. The objectives of this study were threefold: to assess the frequency of forest walking among Japanese residents of Nagoya, a large city; to evaluate differences among frequency of forest walking by participants from a large city, an urban area (Shizuoka; 4,666 participants), and a rural area (Yakumo Town, Hokkaido; 397 participants) in previous studies; and to examine factors related to frequency of forest walking. The survey, by self-administered questionnaire, in the major city of Nagoya was conducted between June 2008 and May 2010. In all, 5,158 participants (M/F, 1,466/3,692; mean age ± SD [range], 52.5 ± 10.3 [35–69] years) were included in the analysis. The proportions of frequency of forest walking ≥ once/month and ≥ once/year were 10.9 % (M/F, 15.1/9.3 %) and 46.1 % (51.0/44.1 %), respectively. In multiple logistic regression analysis, significant differences were noted among study sites for the adjusted odds ratio for frequency of forest walking. Overall, the order of highest to lowest frequency of forest walking was Shizuoka > Nagoya > Yakumo. Factors related to frequency of forest walking were common among the three study sites. Higher frequency of forest walking was associated with male sex and older age; the most relevant factor related to frequency of forest walking was its enjoyment level. Further studies will be required to clarify why these factors are related to frequency of forest walking.     

Mediator-type biosensor for real-time wireless monitoring of blood glucose concentrations in fish

We developed a wireless mediator-type biosensor system to rapidly monitor blood glucose concentrations in free-swimming fish. Glucose oxidase was used to determine the glucose concentration, but fluctuations in the oxygen concentrations in the samples affected the measurements. In this work, we used a mediator, a synthetic electron acceptor, as a substitute in the oxidation–reduction reaction, thus ensuring that changes in the concentration of oxygen do not affect the measurements of glucose concentration performed by the sensor. We especially focused on the use of ferrocene as a mediator. The resulting enzyme sensor utilized a Pt–Ir wire (diameter 0.178 mm) as the working electrode and Ag/AgCl paste as the reference electrode. Chitosan was used to fix ferrocene and glucose oxidase to the working electrode. The characteristics of the sensor were tested, and it was found to be capable of measuring glucose independent of the oxygen concentration of the sample. Its reproducibility was also tested, and the results showed that the sensor was suitable for performing measurements in vivo. The sensor was then inserted into to eyeball interstitial sclera fluid in order to wirelessly monitor the glucose concentration in free-swimming fish. Measurements were taken for a total of 48 h. During these measurements, artificial stress was applied to the fish. The glucose concentrations of fish rise with their stress levels. The output current from the sensor gradually increased during the application of stress, which hinted that the stress was monitored by this system.     

Nickel-catalyzed carbonization of wood for coproduction of functional carbon and fluid fuels II: improved fuel quality of oil fraction and increased heating value of gas fraction

The yields and properties of oil and gas fractions coproduced during carbonization of larch wood loaded with Ni 2%, Ni 2%+Ca 1%, and Ni 4% and without catalyst (None) at 700°–900°C were examined to clarify the catalytic effect in terms of conversion into fluid fuels. The net calorific value of oil occurred mainly below 500°C and increased in the order None < Ni 2% < Ni 4% < Ni 2%+Ca 1%, while the yield decreased in this order. The same order held for the production of gases enriched with hydrogen at 500°–700°C. Even above 800°C, markedly promoted evolution of hydrogen took place for all catalyst systems. These observations confirmed the effectiveness of nickel-catalyzed carbonization at 900°C, particularly Ni 2%+Ca 1%, for both upgrading of oil and gaseous fractions, although the quality of oil was not satisfactory. The catalysis of nickel with and without calcium is discussed on the basis of the modified Broid-Shafizadeh scheme, and the scheme was altered to adapt to the high temperature region where oil was no longer produced. Part of this study was presented at the 14th Annual Meeting of the Japan Institute of Energy, Suita, August 2005, and at the 18th Symposium, Session D, of the Materials Research Society of Japan, Tokyo, December 2007