Feline Ubiquitin Fusion Protein Genes

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.     

Effect of relationships among clinical mastitis incidence,reproductive performance,and culling rate on the lifetime of dairy cows at Hiroshima University Farm

Farm managers' decision to cull dairy cows is based on the cows' milk production, history of disorder(s), and reproductive performance, each of which affects dairy cows' lifetime (herd life and productive lifespan). We investigated the relationships among the incidence of clinical mastitis (CM), the reproductive performance, and the culling rate. We also assessed the effects of these relationships on the lifetimes of dairy cows, using the records made before and after the introduction of an automatic milking system (AMS) at Hiroshima University Farm. Milk yield, CM incidence density, and culling rate of dairy cows increased after the AMS introduction. The CM incidence was associated with an elongation of the calving interval in cows with the same parity. CM in the 1st parity might have caused the reductions of the cows' lifetime and their parity at culling. A higher age at first calving (AFC) was associated with an increase in culling rate but did not lead to a significant decrease in lifetime. Investigations of the factors mediating CM in the 1st parity or AFC with CM incidence or culling rate in the later stages might contribute to the control of lifetime of dairy cows.     

Census and effective population sizes of white‐spotted charr (Salvelinus leucomaenis) in a fragmented landscape

Several habitat models have been proposed to predict population size for stream fishes and to guide habitat assessment and monitoring techniques. However, most models do not incorporate the potential advantage of molecular genetic markers. We conducted a field survey and microsatellite DNA analyses to quantify the relationships among genetic diversity, census/effective population size and habitat variables in fragmented populations of white‐spotted charr (Salvelinus leucomaenis). The census population size significantly increased with the stream length, the number of pools and a pool‐riffle sequence index, a proxy for channel‐unit habitat type complexity within reaches. Population density was correlated with the pool‐riffle sequence index only. Genetic diversity and effective population size were not correlated with the habitat variables or census population size. There was a lack of isolation‐by‐distance population structure in the studied populations. Our results suggest that stream length and the number of pools within reaches associated with habitat complexity are the habitat variables that explain the majority of variation in population size of white‐spotted charr. Our findings provide further evidence that census population size per se is a poor indicator of the inclusive genetic diversity within populations in a fragmented landscape.     

Therapeutic effect of anti-TNF-alpha antibody and levofloxacin (LVFX) in a mouse model of enterohemorrhagic Escherichia coli O157 infection

The ability of an anti-TNF-alpha antibody to confer protection against enterohaemorrhagic Escherichia coli (EHEC) O157 was investigated in germfree IQI mice. The use of an antibiotic levofloxacin (LVFX) alone or with the antibody was also studied. Protection included an increase in survival rate. Treatment with the anti-TNF-alpha antibody inhibited the histological signs associated with EHEC infection but did not prevent the colonization of EHEC or production of Shiga toxin (Stx). No clinical signs were observed and EHEC was completely eliminated in the mouse model receiving both anti-TNF-alpha antibody and LVFX. Anti-TNF-alpha antibody suppressed inflammatory cytokine response in the mouse kidney and brain by EHEC infection.     

Appearance of race Pfs:5 of spinach downy mildew fungus, Peronospora farinosa f. sp. spinaciae, in Japan

The races for the causal agent of spinach downy mildew Peronospora farinosa f. sp. spinaciae were identified by inoculation of race-differential cultivars. One isolate was identified as Pfs:5s and the others belonged to a new race. This is the first report of race Pfs:5 and another new race in Japan.     

Change in flower morphology of Torenia fournieri Lind. induced by forchlorfenuron application

We induced various flower morphologies in torenia (Torenia fournieri Lind.) by the application of forchlorfenuron (CPPU). Those morphologies were the combination of four basic morphological changes, the development of serrate petals, incised petals, a paracorolla, and an increased number of floral organs. These morphological changes occurred systematically depending on the floral stage at the time of CPPU application. Serrate petals were induced when CPPU was applied during the stages of corolla development, whereas application at younger stages induced petal incision. The serrate petal margin resulted from preferential proliferation of cells around the vascular bundles, whereas petal incision likely resulted from the lateral outgrowths of petal. A paracorolla was induced at the adaxial petal face when CPPU was applied between the sepal development stage and early corolla development. The paracorolla appears to have arisen from the lateral outgrowths of the stamen. The numbers of stamens, petals, and sepals increased when CPPU was applied at and before the differentiation of sex organs and the corolla. Enlargement of the floral meristem probably caused this increase. Application of N⁶-benzylaminopurine and zeatin did not induce these morphological changes.     

Isolation and characterization of lectins from the AG-D group of binucleate <Emphasis Type="Italic">Rhizoctonia</Emphasis> species

Isolates from 18 anastomosis groups (AGs) of binucleate Rhizoctonia were screened for lectin activity. Eight AGs (AG-B, AG-D, AG-F, AG-G, AG-H, AG-I, AG-R and AG-U) had low to moderate lectin activities. Among these, members of AG-D and AG-I had the highest activity. Partially purified lectins from AG-D preferentially agglutinated human blood type A to type B and O. Mucin and galactose were the most potent inhibitors among the tested carbohydrates. The molecular masses of these lectins ranged from 12.7 kDa for the monomer to 62 kDa for the pentamer type. Proline, alanine, glutamic acid, aspartic acid, leucine, threonine, serine and tyrosine were the major amino acid components of these lectins. Lectins from AG-D were stable at 4–50°C and from pH 6.0 to 10.0. When assayed with isoelectric focusing, these lectins gave bands at pI 9.30. Specificity of lectins from AG-D to galactose and its derivatives suggest a possible recognition role in this fungal species.     

Multiplication of attenuated and virulent porcine parvoviruses in colostrum-deprived, neonatal pigs

Colostrum-deprived, neonatal, 2 days old pigs were inoculated with the attenuated HT-/SK or the virulent 90HS strain of porcine parvovirus (PPV) by the oral or subcutaneous route and sacrificed 2, 4 or 6 days after inoculation. Then, comparison was made on viral multiplication in pigs between the two strains. Pigs inoculated with the HT-/SK strain showed no detectable viremia or HI antibody responses against PPV within 6 days after inoculation. Only in pigs inoculated by the subcutaneous route, a small amount of virus was recovered from the spleen, liver, or mesenteric lymph nodes. These viruses were distinguished from the parental virulent 90HS strain, as examined for rct maker in vitro. When pigs were inoculated with the virulent 90HS strain, viremia appeared in all of them 1 day after inoculation and continued for up to the sacrificed day. Moreover, a considerable amount of virus was also detected from all tissues, including brain, lung, liver, spleen, pancreas, small intestine, and lymph node tissues, in all pigs tested. HI antibodies were first detected 6 days after inoculation.     

Spatial pattern and regeneration dynamics in a temperate Abies–Tsuga forest in southwestern Japan

This article reports the regeneration dynamics of a temperate Abies–Tsuga forest in Kirishima Yaku National Park, southwestern Japan, and examines the influence of species coexistence mediated by gap disturbances on biomass production. All trees taller than 2 m in a 1-ha plot were monitored over four growing seasons. Three growth-form groups occupied different vertical layers. Evergreen conifers and deciduous broad-leaved trees tended to be spatially segregated from evergreen broad-leaved trees, which formed thickets in the understorey. The regeneration of understorey evergreen broad-leaved trees was affected by canopy gaps. The recruitment of conifers and deciduous broad-leaved species was not observed during the four growing seasons. This suggests that regeneration is sporadic and the present environmental conditions are not favorable for these canopy species. The mortality and unsuccessful recruitment of conifers and deciduous trees appeared to cause fluctuations in the productivity of the stand. However, an abundance of canopy gaps accelerates the regrowth of shorter species, and the fluctuation of productivity resulting from the population dynamics of canopy species would be partly mitigated by the regeneration of evergreen understorey species. The horizontal and vertical heterogeneity of the temperate mixed forest was a result of the patch structures of the three growth-form groups. The different regeneration patterns among the three groups, which were driven by interactions of species-specific regeneration niches and disturbance regimes, might be an important factor in maintaining the aboveground productivity in a transitional mixed forest between warm-temperate and cool-temperate zones.     

Age trends in the genetic parameters of wood density and the relationship with growth rates in hybrid larch (Larix gmelinii var. japonica × L. kaempferi) F1

Age trends in variance components and heritability of overall wood density, earlywood and latewood density, and latewood proportion were investigated in 29-year-old trees of 19 full-sib families of hybrid larch (Larix gmelinii var. japonica × Larix kaempferi) F₁. The age–age correlation and optimum selection age for these traits were also estimated and genetic and phenotypic correlations between wood density and radial growth rate were calculated for each growth ring. Intraring wood density data were obtained using X-ray densitometry. The coefficient of additive genetic variance was stable over all ages, whereas the coefficient of environmental variances gradually decreased with increasing age, resulting in increases in heritability estimates with age for overall density. The latewood proportion had the highest heritability estimates at all ages, ranging from 0.44 to 0.66. Overall density and its various components at 28 years of age showed strong genetic correlations with their respective traits at all younger ages. Optimum selection ages for the wood density traits ranged from 8 to 14 years, at which point maximum gain efficiencies per year were obtained. There were negative correlations between wood density and radial growth rate at early ages, although these relationships tended to be weaker with increasing age. These results suggest that selection at a young age is effective for wood density, but particular care must be taken in selecting trees with an improved radial growth rate because rapid growth will result in a low-density wood product, especially in the early growth period.