Activation and penetration in vitro of pig oocytes treated with calcium ionophore

The present study examined the effects of pre-treatment of pig oocytes with different concentrations (0-50 microM) of calcium ionophore A23187 (CaA) on their activation, development and penetration in vitro. Although untreated oocytes were not activated and did not cleave in culture, high proportions of treated oocytes did so and 20% of oocytes developed to the blastocyst stage when treated with 6.25 microM CaA for 2 min. However, these proportions were reduced in a concentration-dependent manner. When inseminated in vitro with 1 x 10(6) spermatozoa/ml, the penetration rate of oocytes treated with 6.25 microM CaA was similar to that of untreated oocytes. However, fewer oocytes treated with 12.5 and 50 microM CaA were penetrated than untreated oocytes. On the other hand, the proportion of monospermy of oocytes treated with 6.25 microM CaA was higher than the values in oocytes not treated or treated with 50 microM CaA. The time required for zona dissolution of oocytes treated with 6.25 and 12.5 microM CaA was not different from that in untreated oocytes, but oocytes treated with 50 microM CaA required a longer time than untreated oocytes, indicating that zona solubility by protease does not reflect penetrability of oocytes in vitro. When oocytes were inseminated with different concentrations (1-10 x 10(6) cells/ml) of spermatozoa, the highest penetration rate was observed at 1 x 10(6) cells/ml in untreated oocytes and a similar result was obtained in oocytes treated with 6.25 microM CaA. There was no difference in the rate of monospermy in untreated oocytes among different concentrations of spermatozoa, but in treated oocytes, higher proportions of monospermy were observed at 0.5-5 x 10(6) than 10 x 10(6) cells/ml. At 1 x 10(6) cells/ml, the proportion of monospermy was higher in treated than untreated oocytes. These results suggest that pre-treatment of pig oocytes with 6.25 microM CaA, an appropriate concentration, inhibits polyspermic penetration in vitro when insemination occurs with spermatozoa at a concentration of 1 x 10(6) cells/ml.     

Physical, chemical, and biological properties of s-allylcysteine, an amino acid derived from garlic.

Physical, chemical, and biological properties of S-allylcysteine (SAC) were investigated. SAC showed stable properties under tested conditions, and its acute/subacute toxicity was very minor in mice and rats (LD(50) value >54.7 mM/kg po; >20 mM/kg ip). The pharmacokinetics of SAC was investigated after oral administration of garlic supplement containing SAC to human volunteers. SAC from garlic consumption was rapidly absorbed from the gastrointestinal tract, however, the half-life and excretion time were more than 10 h and 30 h, respectively.     

Cloning of estrogen receptor-β cDNA of common Japanese conger Conger myriaster and expression of the gene during oocyte development

ABSTRACT:   Estrogen receptor (ER) cDNA was cloned from the liver of common Japanese conger Conger myriaster . Common Japanese conger ER cDNA contains 2543 nucleotides including an open reading frame that encodes 596 amino acids, with a molecular weight of 66.1 kDa. The deduced amino-acid sequence of the common Japanese conger ER shares 85% identity with that of eel Anguilla japonica ER-β, and 39–46% and 53–59% identity with the deduced sequences of ER-α and ER-β of other species, respectively. In a phylogenetic analysis, the common Japanese conger ER was found to cluster with the ER-β of other species. The common Japanese conger ER-β also has six domains (A–F), which are conserved in the nuclear receptor superfamily. The amino-acid sequences in the C and E domains were highly conserved between common Japanese conger ER-β and other ER. However, the other domains showed low homology. In the female common Japanese conger, ER-β mRNA was highly expressed in the liver and pituitary gland. The expression levels of the ER-β gene increased from the oil droplet stage to the primary yolk globule stage and were maintained until the migratory nucleus stage.     

Oral antibiotics enhance antibody responses to keyhole limpet hemocyanin in orally but not muscularly immunized chickens

Recent studies have emphasized the crucial role of gut microbiota in triggering and modulating immune response. We aimed to determine whether the modification of gut microbiota by oral co‐administration of two antibiotics, ampicillin and neomycin, would lead to changes in the antibody response to antigens in chickens. Neonatal chickens were given or not given ampicillin and neomycin (0.25 and 0.5 g/L, respectively) in drinking water. At 2 weeks of age, the chicks were muscularly or orally immunized with antigenic keyhole limpet hemocyanin (KLH), and then serum anti‐KLH antibody levels were examined by ELISA. In orally immunized chicks, oral antibiotics treatment enhanced antibody responses (IgM, IgA, IgY) by 2–3‐fold compared with the antibiotics‐free control, while the antibiotics did not enhance antibody responses in the muscularly immunized chicks. Concomitant with their enhancement of antibody responses, the oral antibiotics also lowered the Lactobacillus species in feces. Low doses of antibiotics (10‐fold and 100‐fold lower than the initial trial), which failed to change the fecal Lactobacillus population, did not modify any antibody responses when chicks were orally immunized with KLH. In conclusion, oral antibiotics treatment enhanced the antibody response to orally exposed antigens in chickens. This enhancement of antibody response was associated with a modification of the fecal Lactobacillus content, suggesting a possible link between gut microbiota and antibody response in chickens.     

Domestication and cereal feeding developed domestic pig‐type intestinal microbiota in animals of suidae

Intestinal microbiota are characterized by host‐specific microorganisms, which have been selected through host‐microbe interactions under phylogenetic evolution and transition of feeding behavior by the host. Although many studies have focused on disease‐related intestinal microbiota, the origin and evolution of host‐specific intestinal microbiota have not been well elucidated. Pig is the ideal mammal model to reveal the origin and evolution of host‐specific intestinal microbiota because their direct wild ancestor and close phylogenetic neighbors are available for comparison. The pig has been recognized as a Lactobacillus‐type animal. We analyzed the intestinal microbiota of various animals in Suidae: domestic pigs, wild boars and Red river hogs to survey the origin and evolution of Lactobacillus‐dominated intestinal microbiota by metagenomic approach and following quantitative PCR confirmation. The metagenomic datasets were separated in two clusters; the wild animal cluster being characterized by a high abundance of Bifidobacterium, whereas the domesticated (or captured) animal cluster by Lactobacillus. In addition, Enterobacteriaceae were harbored as the major family only in domestic Sus scrofa. We conclude that domestication may have induced a larger Enterobacteriaceae population in pigs, and the introduction of modern feeding system further caused the development of Lactobacillus‐dominated intestinal microbiota, with genetic and geographical factors possibly having a minor impact.     

A canine case with cystic meningioma showing miraculous reduction of the cystic lesion

A 12-year-old spayed female Labrador retriever was presented with forebrain signs. Brain MRI revealed a huge cystic lesion with the thickened falx in the frontal region. The brain parenchyma surrounding the lesion showed significant signs of a mass effect and also increased intracranial pressure. However, the dog suddenly became lucid after about two weeks, and an MRI scan one month after the initial study revealed a dramatically shrunken cystic lesion. The dog survived for over a year until it was euthanized for other reasons, and the brain lesion was diagnosed as a cystic meningioma histologically. To the authors’ knowledge, this is the first report that described the reduction of the cystic lesion of a cystic meningioma in dogs.     

A preliminary investigation on microstructural characteristics of interfacial zone between cement and exploded wood fiber strand by using SEM-EDS

The hydration behavior and strength performance of cement mixed with exploded wood fiber strand (WFS) obtained by the water-vapor explosion process have been studied previously. In the current study, the microstructural characteristics of cement–exploded WFS interfacial zone were examined using scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDS). The Ca/Si ratios at the interfacial zones and the elemental compositions of hydration products deposited in the tracheid lumen were investigated. In addition, the morphological differences and compositional variations of hydration products that developed on the wood surfaces were examined. The results revealed that the Ca/Si ratios at the interfacial zones were strongly influenced by the mixture compositions, and that the elemental compositions of the hydration products that filled the tracheid lumen were significantly different from those of the cement paste in the mixtures. Differences in morphology and composition of hydration products at the wood surfaces were also observed to correspond to the different mixture compositions. These characteristics are considered to be directly related to the bond property, and thus, to the mechanical performance of WCM.Part of this report was presented at the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, March 2003     

Horizontal distribution of stolons of a temperate lianaWisteria floribunda DC. and its ecological significance

The spatial distribution of stolons ofWisteria floribunda DC. was investigated, and the ecological significance of clonal growth of this temperate liana is discussed. The study plot (0.15 ha) was located in a secondary deciduous broad-leaved forest dominated byQuercus serrata in central Japan. The tree canopy was almost closed with a mean openness of 4.2%.W. floribunda was dominant among lianas accounting for 86% of their total basal area.W. floribunda stolons of three individuals branched numerous times and intersected among individuals, and showed a large horizontal “web-like” extension on the ground. The total length of stolons was 66.0, 260.2, and 310.6 m, respectively, for each individual. A mean of 25 ramets were established on stolons per individual, and one-third of the ramets obtained a host tree. These results suggested that the expansion of stolons was effective in obtaining host trees forW. floribunda individuals. On the other hand, clonal growth ofW. floribunda did not always contribute to vertical growth of ramets in the study forest, since few ramets reached the forest canopy. Lack of sub-canopy trees in this forest appeared to constrain the vertical growth ofW. floribunda ramets, since sub-canopy trees are thought to bridge lianas from smaller trees to canopy trees.     

Development of room-temperature curing aqueous emulsion-type acrylic adhesive II: effect of monomer composition on the final adhesive strength

In a series of studies, development of a new aqueous emulsion-type adhesive for use with wood or paper, which does not release formaldehyde or volatile organic solvents, was attempted. For the purpose of increasing both initial and final adhesive strengths, we selected a system consisting of acrylic monomers copolymerized with functional monomers such as diacetone acrylamide and some cross-linking agents such as dihydrazide or polymeric methylene diphenyl diisocyanate (pMDI). It was found that dihydrazide cross-linker was effective for both the initial and final adhesive strengths, suggesting contribution of hydrazone bonds to the bonding strengths. Cross-linking with pMDI might be slow and was not important in the initial bonding. However, its cross-linking effect might develop gradually, because the adhesive showed good final strengths using pMDI as a cross-linker.