Phenotypic and functional properties of feline dedifferentiated fat cells and adipose-derived stem cells

It has been reported that mature adipocyte-derived dedifferentiated fat (DFAT) cells show multilineage differentiation potential similar to that observed in mesenchymal stem cells. Since DFAT cells can be prepared from a small quantity of adipose tissue, they could facilitate cell-based therapies in small companion animals such as cats. The present study examined whether multipotent DFAT cells can be generated from feline adipose tissue, and the properties of DFAT cells were compared with those of adipose-derived stem cells (ASCs). DFAT cells and ASCs were prepared from the floating mature adipocyte fraction and the stromal vascular fraction, respectively, of collagenase-digested feline omental adipose tissue. Both cell types were evaluated for growth kinetics, colony-forming unit fibroblast (CFU-F) frequency, immunophenotypic properties, and multilineage differentiation potential.DFAT cells and ASCs could be generated from approximately 1 g of adipose tissue and were grown and subcultured on laminin-coated dishes. The frequency of CFU-Fs in DFAT cells (35.8%) was significantly higher than that in ASCs (20.8%) at passage 1 (P1). DFAT cells and ASCs displayed similar immunophenotypes (CD44⁺, CD90⁺, CD105⁺, CD14^?, CD34^? and CD45^?). Alpha-smooth muscle actin-positive cells were readily detected in ASCs (15.2 ± 7.2%) but were rare in DFAT cells (2.2 ± 3.2%) at P1. Both cell types exhibited adipogenic, osteogenic, chondrogenic, and smooth muscle cell differentiation potential in vitro. In conclusion, feline DFAT cells exhibited similar properties to ASCs but displayed higher CFU-F frequency and greater homogeneity. DFAT cells, like ASCs, may be an attractive source for cell-based therapies in cats.     

Allelic frequencies and association with carcass traits of six genes in local subpopulations of Japanese Black cattle

Marker‐assisted selection (MAS) is expected to accelerate the genetic improvement of Japanese Black cattle. However, verification of the effects of the genes for MAS in different subpopulations is required prior to the application of MAS. In this study, we investigated the allelic frequencies and genotypic effects for carcass traits of six genes, which can be used in MAS, in eight local subpopulations. These genes are SCD, FASN and SREBP1, which are associated with the fatty acid composition of meat, and NCAPG, MC1R and F11, which are associated with carcass weight, coat color and blood coagulation abnormality, respectively. The frequencies of desirable alleles of SCD and FASN were relatively high and that of NCAPG was relatively low, and NCAPG was significantly associated with several carcass traits, including carcass weight. The proportions of genotypic variance explained by NCAPG to phenotypic variance were 4.83 for carcass weight. We thus confirmed that NCAPG is a useful marker for selection of carcass traits in these subpopulations. In addition, we found that the desirable alleles of six genes showed no negative effects on carcass traits. Therefore, selection using these genes to improve target traits should not have negative impacts on carcass traits.     

Colonization of fleshy-fruited plants beneath perch plant species that bear fleshy fruit

We evaluated the contribution of seed dispersal by birds to the colonization of fleshy-fruited plants beneath plants that bear fleshy fruits and were used as perches by birds. Specifically, we investigated the composition of seeds and saplings sampled beneath nine fleshy-fruited perch plant species in a Pinus thunbergii stand on a coastal sand dune in central Japan. Both the numbers and the species numbers of bird-dispersed (pulpless) seeds differed significantly among the perch plant species. Nevertheless, the quantitative compositions of bird-dispersed seeds were similar among the perch plant species because of the significant positive interspecific correlations of species rank order in numbers of bird-dispersed seeds per species. Similar relationships were also found for saplings. For each perch plant species, the numbers of fallen conspecific seeds were significantly higher than or not different from those of heterospecific seeds. Furthermore, the numbers of pulpy seeds that fell directly from the perch plant were significantly higher than those of bird-dispersed conspecific seeds for seven of nine perch plant species. However, the densities of conspecific saplings were significantly lower than those of heterospecific saplings. Most pulpy seeds, therefore, probably do not germinate beneath their mother plants, and conspecific seeds probably accumulate in seed banks or are consumed by herbivores.     

The perivitelline space-forming capacity of mouse oocytes is associated with meiotic competence

Although mouse oocytes progressively acquire meiotic competence during their growth in the ovaries, only half of full-grown oocytes can accomplish meiosis. Two types of full-grown oocytes have been reported on the basis of their chromatin configuration, the surrounded-nucleolus (SN) type and the non-surrounded-nucleolus (NSN) type. Therefore, full-grown oocytes collected from the ovaries of adult animals comprise a heterogeneous population; some oocytes are meiotically incompetent (NSN-type), and some are competent (SN-type). In the present study, we found that full-grown oocytes could be classified into two groups using the criterion of formation of the perivitelline space (PVS) after culture with 3-isobutyl-1-methylxanthine (IBMX) for 1 h. In oocytes with a PVS, actin-filled processes within zona pellucidae originating from cumulus cells were reduced, while they were rich in oocytes without a PVS, suggesting that a reduction in these processes contributes to PVS formation. PVS formation was highly correlated with meiotic competence and SN-type configuration. The results of this study demonstrate that PVS formation is a useful criterion for easily distinguishing between SN- and NSN-type oocytes, without injury to the cells.     

Physiochemical factors affecting the salt tolerance of Echinochloa crus-galli Beauv. var. formosensis Ohwi

The effects of NaCl-induced stress on physiochemical factors such as inorganic cations, proline, malondialdehyde (MDA) and polyamines were investigated in the gramineous weed, Echinochloa crus-galli Beauv. var. formosensis Ohwi ( E. crus-galli ) and rice ( Oryza sativa L. cv. Nipponbare). Growth inhibition at the 2nd leaf stage under salt stress was more severe in rice than in E. crus-galli . Water content in the 2nd leaves was also more severely decreased in rice, indicating that E. crus-galli was more salt-tolerant. After NaCl treatment, Na⁺ accumulated in the 2nd leaves of both plant species but not in their roots. Proline accumulation in the 2nd leaves was significantly higher in salt stressed E. crus-galli than in rice, suggesting the significance of proline production in the salt tolerance of this weed. Content of MDA of the rice increased more greatly with NaCl treatment than that in E. crus-galli . NaCl treatment affected polyamine metabolism of both plant species, but the response of each plant to salt stress was somewhat different, especially in the leaves. Leaf putrescine and spermidine contents were high in non-stressed plants in salt-sensitive rice, although rather lower in E. crus-galli in response to NaCl concentrations. These results indicate that an increase in proline and changes in polyamines relates to the salt tolerance of E. crus-galli .     

Factors affecting changes in a landscape structure dominated by both primary and coppice forests in theFagus crenata forest region of central Japan

In order to clarify the effects of both the social and natural environment on the changes in landscape structure we analyzed landscape elements using aerial photographs taken in different years in the primary and coppice forests in theFagus crenata forest region of central Japan. The present landscape diversity calculated by Shannon's diversity index (H′) appeared to be higher than that observed in photographs from the earlier year. The site with no residential areas and dominated by primary forests had a lower level of landscape diversity than the site which included some residential areas and coppice forests. These results show that the intensity of human activities contributes to changes in landscape diversity. The landscape diversity on private land was higher than that in the national forest, and the changes in landscape structure in the national forest were different from those seen on private land. This suggests that land ownership significantly affects the changes in landscape structure. Topographic factors also affect the changes in landscape structure. Consequently, not only the natural environment (topography), but also the social environment (intensity of human activity and land ownership) strongly influence the changes in landscape structure.     

The assessment of genetic diversity within and among the eight subpopulations of Japanese Black cattle using 52 microsatellite markers

Japanese Black cattle are at risk for genetic homogeneity due to intensive use of a few sires. Therefore, assessment of the actual genetic diversity of this breed is important for future breeding plans. In the present study, we investigated the genetic diversity within and among eight subpopulations of Japanese Black cattle using 52 microsatellite markers. The parameters for genetic diversity of Japanese Black cattle were comparable to those of other cattle breeds, suggesting that the relatively high genetic diversity of the breed. However, upon comparison among the eight subpopulations, the Hyogo subpopulation showed markedly low genetic diversity. The results of the pairwise F_ST values, phylogenetic network and structure analysis indicated that the Hyogo population has remarkably high level of genetic differentiation from other populations, while Yamagata, Niigata, Hiroshima and Kagawa populations have low levels of genetic differentiation. Furthermore, multidimensional scaling plots indicated that individuals in some subpopulations were separated from individuals in the other subpopulations. We conclude that while the overall genetic diversity of Japanese Black cattle is still maintained at a relatively high level, that of a particular subpopulation is significantly reduced, and therefore the effective population size of the breed needs to be controlled by correct mating strategies.     

Relationship between safening activity of dymron and fenclorim to pretilachlor and glutathione S-transferase activity in rice

Dymron [1‐(α,α‐dimethybenzyl)‐3‐(p‐tolyl)urea] and fenclorim (4,6‐dichloro‐2‐phenylpyrimidine) were found to exhibit a safening activity on the growth of rice (Oryza sativa L.) seedlings against pretilachlor [2‐chloro‐2′,6′diethyl‐N‐(2‐propoxyethyl)acetanilide] injury. By pretilachlor treatment at 10^–6 and 10^–5 mol L^–1, the elongation of the third leaves of rice seedlings was reduced by approximately 20 and 40%, and that of the fourth leaves was reduced by approximately 40 and 80%, respectively. Upon the treatment of dymron at 3 × 10^–6 and 10^–5 mol L^–1 in combination with pretilachlor, the growth inhibition was half alleviated in the third leaves, and the length of the fourth leaves was almost recovered from 10^–6 mol L^‐1 pretilachlor injury, and was 20–25% recovered from 10^–5 mol L^–1 pretilachlor injury. Upon the treatment of fenclorim at 3 × 10^–6 and 10^–5 mol L^–1 in combination with pretilachlor, the growth inhibition of rice seedlings was almost alleviated in both the third and the fourth leaves. This result indicated that dymron and fenclorim showed almost the same safening effect on the fourth leaf growth against 10^–6 mol L^‐1 pretilachlor injury, although fenclorim showed higher effects at higher concentrations of pretilachlor. Glutathione S‐transferase (GST) activities in rice seedlings were investigated after being treated with a herbicide and safener. By pretilachlor treatment at 10^–6 and 10^–5 mol L^–1, the GST activity was approximately 32 and 72% increased in roots, respectively, and a little increased (7–13%) in shoots of two‐leaf‐stage rice seedlings. By dymron treatment at 3 × 10^–6?10^–5 mol L^–1, the GST activity was 2–30% increased in roots, but was not increased in shoots. By their combination treatment, the GST activity was almost the same or less than that by treatment with pretilachlor alone. In contrast, by fenclorim treatment alone, the GST activity was 43–52 and 33–45% increased in roots and shoots of rice seedlings, respectively. By the combination treatment of pretilachlor and fenclorim, the GST activity was increased 73–126% in shoots and 101–139% in roots, and was much more increased in both shoots and roots compared with treatment of pretilachlor or fenclorim alone. It was found that dymron showed less effect in increasing the GST activity than fenclorim. It is also suggested that dymron did not increase the GST activity in shoots but did increase it slightly in roots, and showed almost no effect on GST increase by pretilachlor in shoots, or rather reduced the increase in roots. From the above results, fenclorim and dymron may have different mechanisms of safening effects on the protection of rice seedlings against pretilachlor injury.