Distribution of the pores of epithelial basement membrane in the rat small intestine

The distribution and diameter of the pores of epithelial basement membrane in the intestinal villi and the lymph nodules of ileal Peyer's patches were investigated in the rat small intestine by scanning electron microscopy after the removal of the overlying epithelial cells with OsO(4) maceration. In the duodenum, jejunum and ileum, the pores were mainly distributed at the upper three fourths of the villi, but were scarce around the top of the villi. The diameter of some of the pores in the upper three fourths of the villi was larger than that of those in the lower portion. The protrusion of lymphocytes and the cytoplasmic processes of macrophages were also seen at the orifices of the pores. In ileal Peyer's patches, in contrast, pores were densely distributed in the lower one third of the follicle-associated epithelium (FAE) where M cells were mainly seen. Furthermore, these pores were larger than those found in the upper two thirds. Lymphocytes or cytoplasmic processes of macrophages were frequently seen in the lower one third of FAE. These results suggest that the pores at the basement membrane correspond to the passage of the immunocompetent cells which are in contact with M cells or villous columnar epithelial cells and that the abundance of pores is a sign of aggressive interaction between the particular epithelial cells and the immunocompetent cells at the upper three fourths of intestinal villi and the lower one third of FAE in the rat small intestine.     

PKA implicated in the phosphorylation of Cx43 induced by stimulation with FSH in rat granulosa cells

Connexin 43 (Cx43)-mediated gap junctional communication in granulosa cells is crucial for germ line development and postnatal folliculogenesis. We previously showed that follicle-stimulating hormone (FSH) promoted phosphorylation of Cx43 in rat primary granulosa cells. We further identified Ser365, Ser368, Ser369, and Ser373 in the carboxy-terminal tail as the major sites of phosphorylation by FSH, and found that the phosphorylation of these residues was essential for channel activity. In this study, we investigated the protein kinase(s) responsible for FSH-induced phosphorylation. H89, a cyclic AMP-dependent protein kinase (PKA) inhibitor, inhibited FSH-induced phosphorylation both in vivo and in vitro, whereas PD98059, a mitogen-activated protein kinase kinase (MEK) inhibitor, had little effect on the phosphorylation level. Ca2+-dependent protein kinase (PKC) appeared to negatively regulate phosphorylation. Phosphopeptide mapping with or without H89 treatment indicated that PKA could be responsible for phosphorylation of the four serine residues. In addition, the purified catalytic subunit of PKA could phosphorylate the recombinant C-terminal region of Cx43, but not the variant in which all four serine residues were substituted with alanine. These results suggest that FSH positively regulates Cx43-mediated channel formation and activity through phosphorylation of specific sites by PKA.     

The oral administration of thermophile-fermented compost extract and its influence on stillbirths and growth rate of pre-weaning piglets

Food produced via fermentation with mesophilic bacteria has been used to confer health benefits. In contrast, mammalian physiological responses to the intake of thermophile-fermented products have not been thoroughly investigated. We examined the effects of administering a compost extract consisting of fermented marine animals with thermophiles, including Bacillaceae, to pregnant sows and piglets. Retrospective studies were performed on two different swine farms (n=330-1050 sows). The rate of stillbirth was markedly lower in all parities of the compost extract-fed group compared to those of the control group (p≦0.001). Additionally, the birth to weaning period of newborns was significantly shorter (p<0.0001), while the ratio of weanlings per liveborn piglets was increased by more than 6.5% in the compost extract-fed group. Thus thermophiles and their products in the compost extract might promote growth and reduce stillbirths of piglets during the birth to weaning period.     

Identification of weakly beta-hemolytic porcine spirochetes by biochemical reactions, PCR-based restriction fragment length polymorphism analysis and species-specific PCR

We examined the usefulness of PCR-based restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR combined with a newly devised rapid biochemical test using microplates for identifying weakly beta-hemolytic intestinal spirochetes (WBHIS) isolated from pigs. WBHIS strains showing atypical biochemical characteristics were decisively identified at the species level by PCR-RFLP and species-specific PCR. Identification of WBHIS at the species level in routine diagnostic work will certainly contribute to clarifying the pathogenicity of WBHIS.     

Effect of growth on wood properties for Japanese larch (Larix kaempferi): Differences of annual ring structure between corewood and outerwood

To investigate the relations between growth and the wood properties of Japanese larch (Larix kaempferi), six sample trees of varied ages and radial growth were felled and the ring width, ring density, percentage of latewood, and some other factors were determined. There were significant differences in ring density and percentage of latewood between sample trees with vigorous growth and those with poor growth. In corewood the ring density decreased with increasing ring width for all sample trees, whereas in outerwood this trend did not appear. Moreover, the latewood width increased with the increment of ring width only in outerwood, whereas there was almost no change in the corewood. The variation in patterns of ring width, ring density, and percentage of latewood in the radial direction and the relation with height was also studied.Part of this report was presented at the 47th annual meeting of the Japan Wood Research Society, Kochi, April 1997     

Dispersal ability determines the genetic effects of habitat fragmentation in three species of aquatic insect

• 1. The dispersal ability of species and the geographic scale of habitat fragmentation both may influence the extent of gene flow between fragments, but their interactions have rarely been tested, particularly among co‐occurring species.
• 2. Population genetic structures of three species of aquatic insect were compared in streams fragmented by reservoirs and in unfragmented streams in north‐eastern Japan, using 52, 37, and 58 RAPD markers. The three species studied included a strong disperser Cincticostella elongatula (Ephemeroptera: Ephemerellidae), an intermediate disperser Stenopsyche marmorata (Trichoptera: Stenopsychidae), and a weak disperser Hydropsyche orientalis (Trichoptera: Hydropsychidae).
• 3. The patterns of genetic isolation by distance (IBD) supported a priori hypotheses of dispersal ability. The strong disperser (C. elongatula) exhibited significant IBD only at the largest spatial scale studied (among drainages, r=0.50, P<0.01). The intermediate disperser (S. marmorata) showed IBD both within (r=0.22, P<0.01) and among (r=0.45, P<0.01) drainages. The weak disperser (H. orientalis) did not exhibit significant IBD at any scale.
• 4. Pairwise genetic differentiation (θ) indicated that neither the weak disperser nor the strong disperser were genetically differentiated above and below reservoirs when compared with reference sites. This was in contrast to previous results for S. marmorata, for which subpopulations were genetically fragmented across larger (>4.1 km), but not smaller (<2.9 km) reservoirs.
• 5. We suggest that intermediate dispersers, i.e. those at equilibrium between migration and genetic drift within drainages, are more likely to be affected by fragmentation than either strong or weak dispersers. Intermediate dispersers could therefore be used as indicator species in studies aimed at detecting the effects of distance between habitat fragments (e.g. reservoir size) for conservation planning. Copyright © 2010 John Wiley & Sons, Ltd.

     

Effects of olive leaf powder supplemented to fish feed on muscle protein of red sea bream

Olive leaf is known to have the high polyphenol content of 6–9% in dry weight. We investigated the effects of olive leaf powder (OLP) supplemented to fish feed on muscle protein of red sea bream (Pagrus major). Fish reared with feed containing 8% OLP for 40 days had 1.4 times higher myofibril content and 2.2 times higher acid-soluble collagen content than fish reared with control feed for the same period. On the other hand, sarcoplasmic protein content and collagenase activity of the muscle were almost the same between the control fish and OLP-diet fish. Microstructure observation of fish muscle showed that OLP-diet fish has more rigid endomysium structure than that of the control-diet fish. Since collagen fiber in endomysium is responsible for the texture of the muscle, feeding OLP to aquaculture fish will lead to a harder muscle texture. The present study suggests that OLP is a useful feed additive to enhance the texture of aquaculture red sea bream muscle through strengthening of the collagen structure in the muscle.     

Novel SNP in 5' flanking region of EDG1 associated with marbling in Japanese Black beef cattle

Marbling, defined by the amount and distribution of intramuscular fat, is an economically important trait of beef cattle in Japan. The endothelial differentiation , sphingolipid G-protein-coupled receptor , 1 ( EDG1 ) gene has been considered as a positional functional candidate for the gene responsible for marbling. We have recently reported that 2 single nucleotide polymorphisms (SNPs), c.-312A > G in the 5' untranslated region (UTR) and c.*446G > A in the 3' UTR in EDG1 were associated with marbling in Japanese Black beef cattle, but this was not functional and a causal mutation for marbling. In the present study, we detected 2 novel SNPs, referred to as g.1475435G > A and g.1471620G > T , in the 5' flanking region of the EDG1 between low-marbled and high-marbled steer groups, which were previously shown to have EDG1 expression differences in musculus longissimus muscle. The g.1475435G > A SNP seemed not to segregate in Japanese Black beef cattle. The g.1471620G > T SNP was associated with the predicted breeding value for beef marbling standard number by the analyses using Japanese Black beef cattle population. Based on these findings, we hypothesized that the g.1471620G > T SNP might have an impact on EDG1 expression and also marbling.     

Tree-ring-width chronology of Larix gmelinii as an indicator of changes in early summer temperature in east-central Kamchatka

We developed a 378-year tree-ring-width chronology based on 110 core samples from 55 individual trees of Larix gmelinii (Rupr.) Rupr. growing in a wide open forest close to the tree line in the Kronotsky National Park. Reflecting strong climatic control over tree growth not only within the study area but also more extensively over central Kamchatka, our chronology was well correlated with those from other larch sites. Response analysis with 10-day mean temperature revealed that the ring width was primarily controlled by the temperature of early summer, i.e., of late May through late June (40 days). While the regression models for a formal reconstruction failed to pass stringent verification tests commonly used in dendroclimatology, the relationship between tree growth and climate was statistically significant and credible. We therefore used our chronology as a proxy of early summer temperature. The chronology shows a cool period from the 1660s until the 1680s, followed by gradual warming until ca. 1800, then by a slight cooling trend extending to ca. 1910, and a warming trend continuing up to the present, with decadal fluctuations throughout the chronology. The warming trend found in our chronology over the twentieth century is generally consistent with the ones commonly appearing in higher latitudes.     

Reproductive performance and expression of imprinted genes in somatic cell cloned boars

To assess the performance of boars derived by somatic cell cloning, we analyzed various aspects of their reproductive characteristics and the expression of two imprinted genes. Cloned boars (cloned Duroc × Jinhua) were analyzed for birth weight, growth rate, age at first ejaculation, semen characteristics and fertility, in comparison with naturally bred control boars of the same strain. The expression of imprinted genes was analyzed using the microsatellite marker SWC9 for the paternally expressed gene insulin‐like growth factor ‐2 (IGF2) and with single nucleotide polymorphisms (SNPs) for the gene maternally expressed 3 (MEG3). The cloned boars had high production of semen and were nearly equal in level of fertility to conventional pigs; they showed similar characteristics as naturally bred boars of the same strains. The expression of IGF2 was partially disturbed, but this disturbed expression was not linked to a change in developmental fate or reproductive performance. These results indicate that use of cloned boars could be highly effective for proliferation of pigs with desirable characteristics, preservation of genetic resources and risk reduction against epidemic diseases, such as foot‐and‐mouth disease, through storage of somatic cells as a precautionary measure for use in regenerating pig populations after a future pandemic.