Phosphorylation of inositol 1,4,5-triphosphate receptor 1 during in vitro maturation of porcine oocytes

During fertilization in mammalian species, a sperm-induced intracellular Ca²⁺ signal ([Ca²⁺]_i) mediates both exit of meiosis and oocyte activation. Recently, we demonstrated in mouse oocytes that the phosphorylation levels of inositol 1,4,5 trisphosphate receptor type1 (IP₃R1), the channel responsible for Ca²⁺ release and oscillations during fertilization, changed during maturation and fertilization. Therefore, we examined the expression and phosphorylation of IP₃R1 during in vitro maturation of pig oocytes. Here, our present study shows that expression of IP₃R1 protein did not change during maturation, although the phosphorylation status of the receptor, specifically at an MPM-2 epitope, did. We found that while at the beginning of maturation IP₃R1 lacked MPM-2 immunoreactivity, it became MPM-2 reactive by 24 h and reached maximal reactivity by 36 h. Interestingly, the acquisition of MPM-2 reactivity coincided with the activation of p34^cdc2 kinase and mitogen-activated protein kinase (MAPK), which are involved in meiotic progression. Following completion of maturation, inactivation of MAPK by U0126 did not affect IP₃R1 phosphorylation, although inactivation of p34^cdc2 kinase by roscovitine dramatically reduced IP₃R1 phosphorylation. Neither inhibitor affected total expression of IP₃R1. Altogether, our results show that IP₃R1 undergoes dynamic phosphorylation during maturation and this might underlie the generation of oscillations at fertilization.     

Relationship between coastal water properties and adult return of chum salmon (Oncorhynchus keta) along the Sanriku coast,Japan

We investigated the relationship between adult (age‐4) return rates for chum salmon (Oncorhynchus keta) originating from the Iwate Prefecture, Japan, and coastal environmental conditions during their early ocean life in coastal residency. We analyzed distributions of water properties via intensive hydrographic observations using a conductivity–temperature–depth profiler. Both the return rates and water properties vary strongly over interannual time scales. We found that the time when the return rate decreased drastically corresponded well to the time when the frequency of warm waters in the coastal residency increased.     

Metabolic discrimination of mango juice from various cultivars by band-selective NMR spectroscopy

NMR-based metabolic analysis of foods has been widely applied in food science. In this study, we performed discrimination of five different mango cultivars, Awin, Carabao, Keitt, Kent, and Nam Dok Mai, using metabolic analysis with band-selective excitation NMR spectra. A combination of unsupervised principal component analysis (PCA) with low-field region (1)H NMR spectra obtained by band-selective excitation provided a good discriminant model of the five mango cultivars. Using F(2)-selective 2D NMR spectra, we also identified various minor components in the mango juice. Signal assignment of the minor components facilitated the interpretation of the loading plot, and it was found that arginine, histidine, phenylalanine, glutamine, shikimic acid, and trigonelline were important for classification of the five mango cultivars.     

Neutropenia associated with osteomyelitis due to Hepatozoon canis infection in a dog

A 4-year-old, intact male Shiba dog was referred to Yamaguchi University Animal Medical Center, Yamaguchi, Japan, for the following complaints: anorexia, lethargy, intermittent fever, gingival bleeding and abdominal purpura. The dog presented with persistent neutropenia. Histopathological examination of a bone marrow sample revealed round to oval structures that resembled Hepatozoon micromerozoites and formed a "wheel-spoke" pattern. Furthermore, mature neutrophils were observed around these structures. PCR and sequencing using bone marrow aspirate confirmed Hepatozoon canis (H. canis) infection. These findings suggest that the neutropenia observed in this case was associated with osteomyelitis due to H. canis infection. This is the first report of neutropenia associated with H. canis infection. H. canis infection can be included in the differential diagnosis in canine cases of neutropenia in areas where the disease is endemic.     

Characterization of monoclonal antibodies against canine P-selectin glycoprotein ligand-1 (PSGL-1)

Thirteen different monoclonal antibodies against canine P-selectin glycoprotein ligand-1 (cPSGL-1) were obtained by immunization of rats with cells of a canine lymphoma cell line (Ema). O-sialoglycoprotein endopeptidase treatment of Ema cells showed that all of these antibodies recognized O-glycosylated peptides of canine PSGL-1. Experiments using deletion or point mutants of cPSGL-1 indicated that these antibodies could be categorized into several groups based on their cPSGL-1 recognition characteristics. These anti-cPSGL-1 monoclonal antibodies will be useful for analysis of the canine P-selectin and PSGL-1 system.     

Molecular detection of Anaplasma phagocytophilum in cattle and Ixodes persulcatus ticks

The tick-borne pathogen, Anaplasma phagocytophilum (A. phagocytophilum), the causative agent of human granulocytic anaplasmosis (HGA), is increasingly becoming a public health concern as an aetiological agent for emerging infectious disease. We found A. phagocytophilum infection in a pooled sample of field-collected Ixodes persulcatus (I. persulcatus) ticks from one district in Hokkaido, Japan. Thus, to further investigate the prevalence in field-collected ticks, we used PCR assays targeting the A. phagocytophilum gene encoding 44 kDa major outer membrane protein (p44) for screening of I. persulcatus ticks and samples from cattle from pastures. Out of the 281 I. persulcatus ticks, 20 (7.1%) were found to harbor A. phagocytophilum DNA. The infection rate for A. phagocytophilum in cattle was 3.4% (42/1251). In future studies, it will be necessary to investigate effects of the infection in order to understand its pathogenesis of A. phagocytophilum in domestic animals.     

Increased total nitrogen content of poultry manure by decreasing water content through composting processes

Microbial mineralization of urea and uric acid in poultry litter can lead to loss of nitrogen (N) content and its value as a fertilizer. To minimize the loss of N in the composting processes, controlling the water content in litters is a key to reduce the mineralization processes of N compounds. The N content of litter may be influenced by diets, hen age and the type of poultry houses used. The objectives of the present study were i) to determine the relationship between the water content and the decomposition rate of uric acid in poultry litter and ii) to investigate the effect of hen age and crude protein (CP) percentages in diets on the N content of poultry litter. A layer feeding trial was conducted in two poultry farms with windowless and open-floor houses. An incubation study of poultry litter was performed under different levels of water content. Our study found that the diet CP percentage (16.5–18%) and the growth stage of laying hens did not have a significant effect on the amount of total N (52–56?g?kg^?1) and uric acid-N (26–31g?kg^?1) in fresh litters. At the 7^th day of litter incubation study, the concentration of uric acid-N was 22 g kg^?1 in litters with a water content of 35%, whereas it further decreased to less than 1.3 g kg^?1 in litters with a water content of 55% and higher levels. The decomposition rate of uric acid-N in litter was 0.3–3.1g?kg^?1?day^?1 in the windowless house and 3.1–7.1g?kg^?1?day^?1 in the open-floor house. Decomposition of uric acid in litters was positively correlated to the litter moisture content that is controlled to be lower in windowless houses (40–50%) than in open-floor houses (55–80%) during the composting period. Our study suggests that the use of windowless houses for layer chicken production is effective for producing poultry manure with a high N content.     

Two forms of cytochrome P450 cDNA from 3-methylcholanthrene-treated European eel Anguilla anguilla

ABSTRACT:     The cytochrome P450 (CYP) represents a large group of microsomal monooxygenases that catalyze drugs as well as a host of lethal environmental contaminants such as dioxins, leading to either detoxification and excretion from the animal or generation of carcinogenic intermediates. In the present study two forms of cDNA were cloned (Eu MC1 and Eu MC2) for European eel CYP1A genes by polymerase chain reaction (PCR) techniques. The cDNA of Eu MC1 was 3368 bp long coding 521 amino acid residues, and that of Eu MC2 was 2464 bp long coding 517 amino acid residues. Identities of deduced amino acid sequences between Eu MC1 and Japanese eel CYP1A1 and that between Eu MC2 and the second form of Japanese eel CYP1A were 98% and 97%, respectively, showing decisively that Eu MC1 and Eu MC2 are orthologous to Japanese eel CYP1A1 and the second form of CYP1A, respectively. A striking difference between the two eel species was that the Eu MC1 peptide was two amino acid residues longer than that of the Japanese eel CYP1A1. Existence of two loci of CYP1A in Japanese and European eels may suggest that the two forms of CYP1A exist widely among the eel species, because the divergence between the two eel species has been shown to be close to the basal divergence among eels. The identities in CYP1A may help to estimate genetic distance between European and Japanese eels.     

Oncolytic reovirus synergizes with chemotherapeutic agents to promote cell death in canine mammary gland tumor

The oncolytic effects of reovirus in various cancers have been proven in many clinical trials in human medicine. Oncolytic virotherapy using reovirus for canine cancers is being developed in our laboratory. The objective of this study was to examine the synergistic anti-cancer effects of a combination of reovirus and low doses of various chemotherapeutic agents on mammary gland tumors (MGTs) in dogs. The first part of this study demonstrated the efficacy of reovirus in canine MGTs in vitro and in vivo. Reovirus alone exerted significant cell death by means of caspase-dependent apoptosis in canine MGT cell lines. A single injection of reovirus impeded growth of canine MGT tumors in xenografted mice, but was insufficient to induce complete tumor regression. The second part of this study highlighted the anti-tumor effects of reovirus in combination with low doses of paclitaxel, carboplatin, gemcitabine, or toceranib. Enhanced synergistic activity was observed in the MGT cell line treated concomitantly with reovirus and in all the chemotherapeutic agents except toceranib. In addition, combining reovirus with paclitaxel or gemcitabine at half dosage of half maximal inhibitory concentration (IC₅₀) enhanced cytotoxicity by activating caspase 3. Our data suggest that the combination of reovirus and low dose chemotherapeutic agents provides an attractive option in canine cancer therapy.     

Estimation of the Gentan probability using the price of logs in private plantation forests

The Gentan probability is defined as the probability that a newly planted forest stand will be harvested at a certain age. In this study, the Gentan probability was estimated using the price of logs, in accordance with the notion that the probability is influenced by economic factors. In order to estimate the Gentan probability, we adopted a probability density function based on the forest owners’ decision-making process, which consisted of a decision criterion, a decision criterion function and a criterion threshold. The price of logs was employed for the criterion, and the decision criterion function was defined as the value of the forest stand based on the price of logs. Moreover, the mechanism of changes in the threshold was formulated based on the random walk concept. The parameters of the suggested Gentan probability were calibrated to fit the harvesting tendencies observed in the past. Finally, the Gentan probability distributions estimated using the prices of logs over the past 20 years were compared with those estimated using felling age means and variances. The two distributions were closely correlated during most years. The proposed Gentan probability model takes advantage of short-term yield predictions such as an analysis of forest owners’ reactions to the current price changes.