Synthesis and biological activity of furanyl anti-juvenile hormonal compounds

Twenty-one synthetic compounds, containing one or more furan rings, were demonstrated to possess anti-juvenile hormone (AJH) activity as evidenced by their induction of premature metamorphosis in the milkweed bug, Oncopeltus fasciatus (Dallas) by contact, topical application or fumigation. The ED₅₀ of the four most active analogs required to induce precocious metamorphosis from 3rd-instar nymphs by residue contact in a Petri dish compared favorably with that of precocene II (6,7-dimethoxy-2,2-dimethyl 2H-chromene) a naturally occurring phytochemical AJH. Precocious metamorphosis was fully reversible by co-treatment with juvenile hormone (JH III) or JH analogs, demonstrating that the observed AJH activity resulted from an induced deficiency of juvenile hormone.     

Development of Expressed Sequence Tag (EST)-based Cleaved Amplified Polymorphic Sequence (CAPS) markers of tea plant and their application to cultivar identification

To develop cleaved amplified polymorphic sequence (CAPS) markers for cultivar identification of the tea leaf, 5 primer pairs designed on the basis of genes that encode proteins related to nitrogen assimilation and 26 primer pairs based on expressed sequence tag (EST) sequences of the root of tea plant were screened. From combinations of primer pair and restriction enzyme that showed polymorphism among tea plants, 16 markers were selected and applied to DNA fingerprinting of Japanese tea cultivars. Sixty-three cultivars, except for a bud sport (Kiraka) and its original cultivar (Yabukita) and a pair that was the progeny of the same crossing parent (Harumoegi and Sakimidori), were distinguished from one another. By combining the 16 markers with previously developed CAPS markers and observing the physical appearance, 67 cultivars were distinguishable. The cultivars involve approximately 95% of total tea cultivating area in Japan; therefore, about 95% of tea leaves produced in Japan can be authenticated by labeling their cultivars.     

The solubilization of myofibrillar proteins of vertebrate skeletal muscle in water

Myofibrillar proteins of vertebrate skeletal muscles are insoluble in solutions of ionic strength that approximate physiological conditions. We established a method to solubilize more than 80% of chicken breast muscle myofibrillar proteins in water for the use of meat as a source of food protein. SDS‐polyacrylamide gel electrophoretic patterns of water‐soluble myofibrillar proteins demonstrated that all identified myofibrillar proteins except connectin/titin were soluble in water. A part of α‐actinin was released from myofibrils by repeated washing with 2.5 mmol/L NaCl and 5 mmol/L L‐histidine solution, and subsequent destruction of connectin/titin in washed myofibrils by ultrasonication resulted in solubilization of a large fraction of chicken breast muscle myofibrillar proteins in water. Myofibrillar proteins of chicken leg, pork loin, beef shoulder loin, and lamb were also solubilized in water using this procedure.     

Confirmation and characterization of murine body weight QTLs, Bwq1 and Bwq2, identified in C57BL/6J x KK-Ay/a F2-Ay/a mice

Body weight quantitative trait loci (QTLs), Bwq1 and Bwq2, identified previously in C57BL/6J x KK-Ay/a F2-Ay/a mice, were further confirmed and characterized. Body weight measurement was done from 21 days after birth (Day 21) through Day 100, at 10-day intervals. Bwq1 was statistically significant only on Days 40, 50, and 60, whereas Bwq2 was statistically significant on and after Day 40. When body weight gain (WG) between two successive weight measurements was evaluated, both Bwq1 and Bwq2 were statistically significant only for WG between Days 30 and 40. The results suggest that variations in body weight among F2-Ay/a individuals in later life have been determined by variations in WG during the period shortly after weaning. The results also suggest that Bwq1 is related to increased body weight in the KK strain, because the effect of Bwq1 on the body weight is observed not only in F2-Ay/a, but also in F2-a/a. On the other hand, it is suggested that Bwq2 is related to enhanced obesity caused by Ay mutation and therefore is a genetic modifier that specifically interacts with the Ay allele, because the effect of Bwq2 is only observed in F2-A y/a. There are two candidate genes, Pparg and Hrh1, which are located near the 95% confidence interval of Bwq2, and which are expressed in the adipose tissue; however, we could not find any nucleotide differences in both cDNAs between KK and C57BL/6J strains.     

Development of an Efficient Method for Genotyping at the gd Locus in NC/Sgn Mice Based on PCR-RFLP Analysis

Growth deficit (gd) is a recessive mutation that occurs spontaneously in the inbred NC/Sgn mouse strain. Because homozygotes (gd/gd) of both sexes are sterile, they must be produced by mating putative heterozygous carriers (+/gd) whose phenotypes are essentially the same as those of wild-type +/+ mice. The objectives of this study were to develop an efficient method that distinguished a gd allele from a wild-type allele and, if possible, to identify nucleotide substitutions responsible for the gd mutation. The location of the gd locus was estimated to be at 58.3 Mbp on chromosome 4, over which Musk is located. An A-to-G base substitution, which resulted in an M826V amino acid exchange, was identified within a tyrosine kinase domain of Musk. This base substitution disrupted a recognition site for NlaIII; this allowed for discriminating the gd allele from the wild-type allele using PCR-RFLP analysis. When 130 (C57BL/6J × NC/Sgn-gd) F(2) mice were genotyped by PCR-RFLP analysis, all 32 growth-retarded F(2) mice were judged to have the gd/gd genotype. Musk mutations are known to cause congenital myasthenia, which is accompanied by growth retardation, postnatal lethality, and development of a hunchback. These were the typical phenotypes of gd/gd mutants. Although we cannot rule out the possibility that the neighboring genes around the Musk locus are related to the gd phenotype, gd could possibly be classified as a mutant allele of Musk.     

Relative shoot height and irradiance and the shoot and leaf properties of Quercus serrata saplings

Effects of relative shoot height and irradiance on shoot and leaf properties of Quercus serrata Thunb. saplings growing in the understory and in gaps were investigated. Photosynthetic photon flux (PPF) at the location of the shoot relative to that in the open (relative PPF; rPPF) and the height of the shoot base relative to tree height (relative height; rHeight) were measured for all current-year shoots of each sapling. Current-year shoot properties (length, leaf area, number of daughter shoots) and mortality, and leaf properties (mass per area (LMA) and nitrogen content per area (N(area))) were examined in relation to rPPF and rHeight. N(area) was used as a proxy for area-based assimilative capacity. Shoot length, leaf area per shoot and number of daughter shoots increased with increasing rHeight, especially in well-lit conditions. Shoot mortality decreased with increasing rHeight and rPPF. Both LMA and N(area) were positively correlated with rPPF, but not rHeight.     

Gastrointestinal motility modulation by stress is associated with reduced smooth muscle contraction through specific transient receptor potential channel

Excessive stress response causes disability in social life. There are many diseases caused by stress, such as gastrointestinal motility disorders, depression, eating disorders, and cardiovascular diseases. Transient receptor potential (TRP) channels underlie non-selective cation currents and are downstream effectors of G protein-coupled receptors. Ca²⁺ influx is important for smooth muscle contraction, which is responsible for gastrointestinal motility. Little is known about the possible involvement of TRP channels in the gastrointestinal motility disorders due to stress. The purpose of this study was to measure the changes in gastrointestinal motility caused by stress and to elucidate the mechanism of these changes. The stress model used the water immersion restraint stress. Gastrointestinal motility, especially the ileum, was recorded responses to electric field stimulation (EFS) by isometric transducer. EFS-induced contraction was significantly reduced in the ileum of stressed mouse. Even under the conditions treated with atropine, EFS-induced contraction was significantly reduced in the ileum of stressed mouse. In addition, carbachol-induced, neurokinin A-induced, and substance P-induced contractions were all significantly reduced in the ileum of stressed mouse. Furthermore, the expression of TRPC3 was decreased in the ileum of stressed mouse. These results suggest that the gastrointestinal motility disorders due to stress is associated with specific non-selective cation channel.     

OSTEORADIONECROSIS AND RADIATION INDUCED BONE TUMORS FOLLOWING ORTHOVOLTAGE RADIATION THERAPY IN DOGS

Osteoradionecrosis and radiation-induced bone tumors are rare complications of radiation therapy. Little information regarding these complications is available in veterinary medicine. We characterized these complications and investigated risk factors in 119 dogs (122 sites) that received definitive orthovoltage radiation therapy to appendicular sites. Long-term survival was expected in all dogs. The complications of interest were osteoradionecrosis and secondary bone tumor, evaluated radiographically, histopathologically, or both. Complication rates were estimated using the Kaplan–Meier product-limit method, and Fisher's exact test or chi-square test was used to compare the complication rate. The median survival time was 1405 days, with median follow-up duration of 657 days. There were 10 radiation-induced bone tumors and five radiation-induced fractures, with two dogs developing both, for an overall complication rate of 11%. The latent period ranged from 1.2 to 6.4 years for osteoradionecrosis and from 2.6 to 8.7 years for radiation-induced bone tumor. Complications were significantly higher in the humerus ( P <0.0001), and in dogs younger than 7 years ( P =0.014). Similar assessment of complications in dogs irradiated with megavoltage photons or electrons are needed.     

Keratometry in normal cats: a cross-sectional study in Japan using an automated handheld keratometer

Keratometry was performed in 73 domestic cats of varied signalment in Japan using an automated handheld keratometer. The mean corneal curvature radius was significantly lower for cats younger than 1 year than for those older than 2 years (8.04 mm vs. 8.80−8.99 mm, P<0.01). The radius was significantly greater in males than in females among the cats older than 11 years (9.22 mm vs. 8.84 mm, P=0.01), while the age distributions of the males and females were similar. Corneal astigmatism did not significantly differ across the gender and age groups. The predictability of the corneal curvature and astigmatism was approximately 41−43% and less than 3%, respectively, as a function of age and bodyweight. The results highlighted some age- and sex-related keratometric variations in domestic cats in Japan.     

Annual changes in the hatching period of the dominant cohort of larval and juvenile ayu Plecoglossus altivelis altivelis in the Shimanto Estuary and adjacent coastal waters during 1986–2001

ABSTRACT:   Larval and juvenile ayu from seven year-groups (1986, 1987, 1992, 1995, 1996, 1999 and 2000) were collected in the Shimanto Estuary and adjacent coastal waters. The present study focuses on the variation among year-groups in the hatching period estimated by age determination using the otolith. The hatching period of the dominant cohorts in the 1986, 1987 and 1992 year-groups occurred from late October to mid-November. A delay of the hatching period of the dominant cohort was observed from the 1995 year-group, which occurred in late November to early December, and the dominant periods in the 1996, 1999 and 2000 year-groups were observed from early to late December. It appears that the delay of the hatching period of dominant cohorts was not due to a delay of spawning, but a high mortality of early hatched larvae. The water temperature in autumn in the coastal waters adjacent to the Shimanto River has tended to rise over the past 20 years and this trend was especially notable in the late 1990s. The delay in the hatching period of the dominant cohort observed from the 1995 year-group was likely to be related to the rise in seawater temperatures in autumn.