Temporal dynamic changes in synthesis of chondroitin sulfate isomers in canine articular chondrocyte culture

To investigate temporal dynamic changes in the synthesis of chondroitin 6-sulfate (CS6) and chondroitin 4-sulfate (CS4) in vitro, normal articular cartilage of femoral heads was harvested from three dogs. Chondrocytes were isolated and cultured in alginate microspheres for 21 days. On days 7, 14 and 21, DNA content was quantified by fluorometric assay using Hoechst 33258. On days 14 and 21, proteoglycans were extracted, and the amounts of CS6 and CS4 were quantified after chondroitinase ABC digestion using capillary electrophoresis. The DNA content and amounts of CS6 and CS4 increased during the culture period. The amounts of CS6 and CS4 divided by DNA content revealed that the synthesis of CS6 was more up-regulated than CS4.     

Sex and age differences in meat composition of Yeso sika deer (Cervus nippon yesoensis) reared for a short period after capture in the wild

Yeso sika deer captured in winter around Lake Akan in Hokkaido were reared for 8–10 months at Tokyo University of Agriculture in Abashiri. Six 1‐year‐old females and males and six 2‐year‐old or older (adult) females and males were slaughtered and their carcasses were processed. The chemical composition, mineral contents and fatty acid composition of the loin were measured. No marked influence of gender or age was noted in the chemical composition of loin. In the mineral contents, significant differences were noted. The potassium and sulfur contents were lower and the sodium content was higher in adult deer meat (P < 0.05, respectively) and the potassium content was higher in male deer meat (P < 0.05). Arsenic, cadmium or lead were not detected. In the unsaturated fatty acid, a significant interaction was detected (P < 0.05), and it was high in 1‐year‐old female deer meat and low in 1‐year‐old male deer meat. Significant gender or age differences were noted only in the mineral contents in the loin of deer reared for a short period after capture.     

Gene expression profiling in embryonic chicken ovary during asymmetric development

The reproductive system in female birds arises as bilateral asymmetrical anlagen, excluding the birds of prey. Earlier, histological and messenger RNA (mRNA) expression profile studies of several genes related to gonadal sex differentiation in chicken embryos tried to elucidate the query of this asymmetry in a scattered manner. To understand the matter precisely, we have focused on mRNA expression of a cohort of genes (FSHR, CYP19A1, caspase 3, caspase 8) in second half of the embryonic days (E10–E18). The established role of leptin in development of the embryo and its expression in the embryonic ovary also drove us to check leptin receptor (LEPR) expression in the ovary. Increased expression of FSHR and CYP19A1 in the left ovary compared with that in the right ovary was identified (P < 0.05), promoting preferential left ovarian development and functionality. Significant high expression (P < 0.05) of the apoptotic genes in the right ovary were also involved here. Leptin probably has no direct influence on ovarian asymmetry as no significant variation in gonadal mRNA expression of LEPR was observed within the same experimental days. We propose that asymmetric expression of this cohort of genes (FSHR, CYP19A1, caspase 3, caspase 8) leads to the development of dimorphic gonads during embryogenesis.     

One year investigation of the prevalence and diversity of clostridial spores in raw milk from the Tokachi area of Hokkaido

We investigated the seasonal prevalence and diversity of clostridial spores in raw milk from the Tokachi area of Hokkaido. Samples of raw milk were collected quarterly from May 2013 through February 2014. The mean clostridial spore count for the raw milk from 336 milk tankers was 27.6 CFU/100 ml. The clostridial species isolated most frequently from raw milk samples was Clostridium tyrobutyricum. The dominant species was C. tyrobutyricum regardless of the season. The percentage of samples with low spore counts (<10 CFU/100 ml) was highest (60.9%) during winter (February) and lowest (34.5%) in autumn (November). In comparison, the percentage of samples with high spore counts (>100 CFU/100 ml) was highest (5.7%) in autumn (November) and lowest (1.1%) during spring (May).     

The Afferent Venous Vessels to the Liver and the Intrahepatic Portal Distribution in the Fowl

The afferent veins to the liver of 325 adult fowls belonging to ten breeds were investigated by celluloid and neoprene latex casts, and venography. The author elucidates macroscopically the basic pattern of the hepatic portal vein. The right and left portal veins ramify the main branches to the liver, therefore, the intrahepatic distribution is divided into the two portal territories varying in relative size, but this change relates to the development of the medial branch from the right portal vein and the presence of the hepatic fissure.     

Ultrasonic treatment to improve the quality of recycled pulp fiber

The effect of ultrasound on the quality of recycled fibers was investigated. Ultrasound was applied to recycled pulp fiber suspension before ink removal by conventional flotation. The ultrasonic treatment induced an increase in the sedimentation volume of the fiber, which implies that the flexibility and bulkiness of the fiber increase. The water retention value of the ultrasonically treated fiber recovered from the loss caused by the recycling. These facts are due to a beating effect of the treatment. The paper sheets prepared from the ultrasonically treated fiber suspension showed higher sheet density, tensile strength, and brightness than that from an untreated fiber suspension. The process, which consists of ultrasonic treatment for 1 min following flotation deinking, requires about 1.4 times as much energy as the conventional flotation deinking process, but it induced 20% improvement in brightness. The results indicate that the ultrasonic treatment is effective in improving recycled fiber quality.This report was presented in part at the 48th Annual Meeting of the Japan Wood Research Society, Shizuoka, April 1998; and at the 5th Annual Meeting of the Japan Cellulose Society, Kyoto, July 1998     

Effects of elevated atmospheric CO2 concentration on the nutrient uptake characteristics of Japanese larch (Larix kaempferi)

We evaluated the response of Japanese larch (Larix kaempferi Sieb. & Zucc.) to elevated atmospheric CO(2) concentration ([CO(2)]) (689 +/- 75 ppm in 2002 and 697 +/- 90 ppm in 2003) over 2 years in a field experiment with open-top chambers. Root activity was assessed as nitrogen, phosphorus and potassium uptake rates estimated from successive measurements of absorbed amounts. Dry matter production of whole plants was unaffected by elevated [CO(2)] in the first year of treatment, but increased significantly in response to elevated [CO(2)] in the second year. In contrast, elevated [CO(2)] increased the root to shoot ratio and fine root dry mass in the first year, but not in the second year. Elevated [CO(2)] had no effect on tissue N, P and K concentrations. Uptake rates of N, P and K correlated with whole-plant relative growth rates, but were unaffected by growth [CO(2)], as was ectomycorrhizal colonization, a factor assumed to be important for nutrient uptake in trees. We conclude that improved growth of Larix kaempferi in response to elevated [CO(2)] is accompanied by increased root biomass, but not by increased root activity.     

Characterization of an Unknown Region Linked to the Glycoside Hydrolase Family 17 β-1,3-Glucanase of Vibrio vulnificus Reveals a Novel Glucan-Binding Domain

The glycoside hydrolase family 17 β-1,3-glucanase of Vibrio vulnificus (VvGH17) has two unknown regions in the N- and C-termini. Here, we characterized these domains by preparing mutant enzymes. VvGH17 demonstrated hydrolytic activity of β-(1→3)-glucan, mainly producing laminaribiose, but not of β-(1→3)/β-(1→4)-glucan. The C-terminal-truncated mutants (ΔC466 and ΔC441) showed decreased activity, approximately one-third of that of the WT, and ΔC415 lost almost all activity. An analysis using affinity gel containing laminarin or barley β-glucan revealed a shift in the mobility of the ΔC466, ΔC441, and ΔC415 mutants compared to the WT. Tryptophan residues showed a strong affinity for carbohydrates. Three of four point-mutations of the tryptophan in the C-terminus (W472A, W499A, and W542A) showed a reduction in binding ability to laminarin and barley β-glucan. The C-terminus was predicted to have a β-sandwich structure, and three tryptophan residues (Trp472, Trp499, and Trp542) constituted a putative substrate-binding cave. Linker and substrate-binding functions were assigned to the C-terminus. The N-terminal-truncated mutants also showed decreased activity. The WT formed a trimer, while the N-terminal truncations formed monomers, indicating that the N-terminus contributed to the multimeric form of VvGH17. The results of this study are useful for understanding the structure and the function of GH17 β-1,3-glucanases.     

Non-steroidal inhibitors of Drosophila melanogaster steroidogenic glutathione S-transferase Noppera-bo

Insect growth regulators (IGRs) can be developed by elucidating the molecular mechanisms of insect-specific biological events. Because insect molting, and metamorphosis are controlled by ecdysteroids, their biosynthetic pathways can serve as targets for IGR development. The glutathione S-transferase Noppera-bo (Nobo), which is conserved in dipteran and lepidopteran species, plays an essential role in ecdysteroid biosynthesis. Our previous study using 17β-estradiol as a molecular probe revealed that Asp113 of Drosophila melanogaster Nobo (DmNobo) is essential for its biological function. However, to develop IGRs with a greater Nobo inhibitory activity than 17β-estradiol, further structural information is warranted. Here, we report five novel non-steroidal DmNobo inhibitors. Analysis of crystal structures of complexes revealed that DmNobo binds these inhibitors in an Asp113-independent manner. Among amino acid residues at the substrate-recognition site, conformation of conserved Phe39 was dynamically altered upon inhibitor binding. Therefore, these inhibitors can serve as seed compounds for IGR development.     

Genetic parameters for mastitis incidence and its indicators based on somatic cell score for Holsteins in Hokkaido,Japan

The objectives of this study were to estimate the heritability of mastitis incidence and genetic correlations between the mastitis and the somatic cell score (SCS) statistics, and to compare the practicability between different models. We used test‐day records with the mastitis incidence and SCS collected from Holstein cows calving from 1988 to 2015 in Hokkaido, Japan. As indicators of mastitis, the average SCS (avSCS), the standard deviation of SCS (sdSCS), and the maximum SCS (maxSCS) were calculated using test‐day records up to the first 305 days in milk within a lactation. We compared a four‐trait repeatability animal model (MTRP) with a four‐trait multiple‐lactation animal model (MTML). The heritability for mastitis was equal to or lower than 0.05 in all the models. Genetic correlations between lactations with MTML within the same trait were positive and close to 1. With MTRP, the estimated genetic correlations of the mastitis incidence with avSCS, sdSCS, and maxSCS were 0.66, 0.79, and 0.82, respectively. A joint evaluation with SCS statistics is expected to give an extra reliability for mastitis because of high and positive genetic correlations among the traits.