Effect of estradiol‐17β during in vitro growth culture on the growth,maturation, cumulus expansion and development of porcine oocytes from early antral follicles

Growing porcine oocytes from early antral follicles can acquire meiotic and developmental competence under suitable culture conditions, but at lower rates compared to full‐grown oocytes. We postulated that estradiol‐17β (E₂) supported the acquisition of meiotic and developmental competence as well as cumulus‐expansion ability during growth culture. Growing oocytes from early antral follicles (1.2 to 1.5 mm in diameter) were grown in vitro for 5 days in a medium containing 0, 10^?7, 10^?6, 10^?5 or 10^?4 mol/L E₂; after in vitro maturation, 35, 58, 47, 74 and 49% of oocytes matured to metaphase II, 25, 79, 77, 90 and 97% acquired cumulus‐expansion ability, and 23, 54, 63, 89 and 64% were fully surrounded by cumulus cells, respectively. Following maturation, electro‐stimulation was applied to the oocytes grown with 10^?5 mol/L E₂. After 6 days of culture, in vitro‐grown oocytes developed to the blastocyst stage at a rate similar to that for full‐grown oocytes (31% and 40%, respectively). Therefore, we suggest that the use of E₂ during growth culture improves the meiotic and developmental competence of oocytes, cumulus‐expansion ability, and cumulus cell attachment to the oocytes.     

New lactic acid bacterial strains from traditional Mongolian fermented milk products have altered adhesion to porcine gastric mucin depending on the carbon source

Attachment of lactic acid bacteria to the mucosal surface of the gastrointestinal tract is a major property of probiotics. Here, we examined the ability of 21 lactic acid bacterial strains isolated from traditional fermented milk products in Mongolia to adhere to porcine gastric mucin in vitro. Higher attachment was observed with Lactobacillus delbrueckii subsp. bulgaricus strains 6‐8 and 8‐1 than with Lactobacillus rhamnosus GG (positive control). Lactococcus lactis subsp. cremoris strain 7‐1 adhered to mucin as effectively as did strain GG. Heat inactivation decreased the adhesive ability of strains 6‐8 and 8‐1 but did not affect strain 7‐1. The adhesion of strains 6‐8, 7‐1 and 8‐1 was significantly inhibited when the cells were pretreated with periodate and trypsin, indicating that proteinaceous and carbohydrate‐like cell surface compounds are involved in the adhesion of these strains. The adhesion of strain 7‐1 was affected by the type of carbohydrate present in the growth medium, being higher with fructose than with lactose, galactose or xylose as the carbon source. The sugar content of 7‐1 cells grown on various carbohydrates was negatively correlated with its adhesive ability. We provide new probiotic candidate strains and new information regarding carbohydrate preference that influences lactic acid bacterial adhesion to mucin.     

A comparative study of zinc protoporphyrin IX‐forming properties of animal by‐products as sources for improving the color of meat products

The objective of this study was to obtain fundamental data for improving the color of meat products by using animal by‐products. We investigated zinc protoporphyrin IX (ZnPP)‐forming properties of various internal organs from pigs and chickens. ZnPP was formed in the liver, heart and kidney, whereas the porcine spleen and bile, which are involved in the metabolism of heme, did not have ZnPP‐forming properties. The optimum pH values were different among the internal organs and the ZnPP‐forming properties of porcine organs were better than those of chicken organs. The porcine liver showed the greatest ZnPP‐forming properties among all of the internal organs investigated in this study. The optimum pH value for ZnPP formation in the liver was lower than that of skeletal muscle. Oxygen did not inhibit the formation of ZnPP in the liver, unlike in skeletal muscle. Animal by‐products such as the liver have good ability for the formation of ZnPP and might be useful for improving the color of meat products.     

Molecular characteristics of porcine Fas-associated death domain (FADD) and procaspase-8

To reveal the intracellular signal transduction molecules involved in granulosa cell apoptosis in porcine ovarian follicles, we cloned the porcine Fas-associated death domain (FADD), an adaptor protein for the cell death receptor, and procaspase-8, an initiator caspase. Porcine FADD (pFADD) was 636 bp (211 amino acids: aa) long and showed 74.0 and 65.4% homology with human and murine FADD, respectively. Porcine procaspase-8 (pprocaspase-8) was 1,431 bp (476 aa) long and 70.6 and 63.4% homologous with human and murine procaspase-8, respectively. To confirm the apoptosis-inducing abilities, we constructed pFADD and pprocaspase-8 cDNA expression vectors with enhanced green fluorescence protein (EGFP) and then transfected them into human uterine cervix tumor (HeLa-K), human granulosa cell-derived (KGN), murine granulosa-derived tumor (KK1), and porcine granulosa cell-derived (JC410) cells. When pFADD and pprocaspase-8 were overexpressed, cell death was induced in these transfected cells. However when caspase-inhibitor p35 was cotransfected, cell death was inhibited. The pFADD and pprocaspase-8 genes are well conserved, as are the physiological functions of their products.     

Native and introduced Far Eastern daces,genus Tribolodon,in the Tama River system,Japan, inferred from mitochondrial DNA analysis

Fisheries Science - To investigate the species composition and genetic characteristics of Far Eastern daces, genus Tribolodon, in the Tama River, Japan, partial DNA sequences of the mitochondrial...     

Successful treatment of feline hyperadrenocorticism with pituitary macroadenoma using radiation therapy: a case study

A 10-year-old castrated male cat showing behavioral (irritation, prowling, and tumbling) and cutaneous abnormalities such as dermal fragility was diagnosed as hyperadrenocorticism with pituitary macroadenoma, concurrent with insulin dependent diabetes mellitus. Pituitary enlargement (18.0 mm) was observed during magnetic resonance imaging. High endogenous adrenocorticotropic hormone levels (>2,500 pg/ml) were also observed. Although trilostane treatment (5–10 mg/head, daily) was commenced, the clinical signs did not disappear. Insulin and trilostane treatment were discontinued on day 86 after first day of radiation therapy (4 Gy/12 fractions). After radiation therapy, a decreased pituitary tumor size (10.7 mm) was observed on day 301; neurological and dermatological signs exhibited remission. Radiation therapy is the treatment of choice for feline hyperadrenocorticism with pituitary macroadenoma with neurological signs.     

Effects of exogenous methyl jasmonate and light condition on grape berry coloration and endogenous abscisic acid content

Suitable postharvest treatment methods were investigated to improve the color of grape berries. Culture solutions containing jasmonic acid (JA), methyl jasmonate (MeJA), and prohydrojasmon (PDJ) enhanced the skin coloration of grape berries (‘Pione’) harvested at the initial stage of coloration. MeJA vapor treatment under sealed conditions increased anthocyanin accumulation in grape berries (‘AkiQueen’ and ‘Pione’) harvested at the early stage of skin coloration. Furthermore, promoting skin coloration by MeJA vapor treatment was as effective in mature clusters as it was in detached berries. These effects were confirmed in light conditions but not in constant darkness. Our results showed that postharvest MeJA vapor treatment improved skin coloration in grapes. In addition, postharvest treatment with MeJA was found to have no effect on the endogenous abscisic acid content of grape berry skins. Therefore, we suggest that MeJA vapor treatment can be a useful and labor-saving method for the horticultural industry.     

Overexpression of the Barley Nicotianamine Synthase Gene HvNAS1 Increases Iron and Zinc Concentrations in Rice Grains

In humans, iron (Fe) and zinc (Zn) deficiencies result in major worldwide health problems. Transgenic technologies to produce Fe- and Zn-biofortified rice varieties offer a promising potential solution. Nicotianamine, the precursor of phytosiderophores, chelates Fe²⁺ and Zn²⁺ and plays an important role in transporting these metals to both vegetative and reproductive organs within the plant. Our objective was to increase Fe and Zn contents in rice grains by overexpressing the barley nicotianamine synthase gene HvNAS1. HvNAS1-overexpressing transgenic rice showed increased HvNAS1 expression and subsequent increases in endogenous nicotianamine and phytosiderophore content in shoots, roots, and seeds. Fe and Zn concentrations in polished T₁ seeds from transgenic plants increased more than three and twofold, respectively; Fe and Zn concentrations also increased in both polished and brown T₂ seeds. These results suggest that the overproduction of nicotianamine enhances the translocation of Fe and Zn into rice grains.     

Genetic identification and morphology of naturally spawned eggs of the Japanese eel Anguilla japonica collected in the western North Pacific

Eggs of the Japanese eel Anguilla japonica collected in the western North Pacific were identified by onboard species-specific polymerase chain reaction (PCR) and DNA nucleotide sequencing after the cruise. Fish eggs of various species were collected by large plankton net tows at 12 stations along the southern part of the West Mariana Ridge on 19–25 May 2009. A total of 43 fish eggs were distinguished morphologically as possibly being of A. japonica. Thirty-one of those were analyzed by PCR, which included 15 eggs collected at 12°50–55′N, 141°15–20′E (in 5 tows) that showed positive results. The 16S ribosomal RNA (rRNA) gene sequences of eggs determined after the cruise indicated that 31 A. japonica eggs had been collected. The remaining eggs were of mesopelagic eel species (Serrivomeridae and Derichthyidae), or unidentified species. The morphological characteristics of the A. japonica eggs were consistent with those of artificially spawned eggs, except they had a slightly larger diameter. The egg diameter range did not overlap with those of mesopelagic eels of the Serrivomeridae, which often spawn in the same area as A. japonica. These results suggest that egg diameter and embryo shape can be used to morphologically identify naturally spawned A. japonica eggs.