Singlet diradicals: from transition states to crystalline compounds

Singlet diradicals are usually not energy minima. As observed by femtosecond spectroscopy, they readily couple to form final sigma bonds. Substituent effects allow lifetimes to increase into the microsecond range. Taking advantage of the properties of hetero-elements, a diradical has been prepared that is indefinitely stable at room temperature. The availability of diradicals that can be handled under standard laboratory conditions will lead to further insight into their chemical and physical properties, raising the likelihood of practical applications, especially in the field of molecular materials such as electrical conductors and ferromagnets.     

Development of thyroid gland and changes in thyroid hormone levels in Leptocephali of Japanese Eel (Anguilla japonica)

Development of the thyroid gland of the Japanese eel (Anguilla japonica) was studied with the use of tank-reared fish. A single thyroid follicle was first found in larvae at 29 days post hatching (dph), total length (TL) 12 mm. Until reaching 25 mm in TL (100 dph), leptocephali had one or two follicles per individual. The inner colloid of the follicles was weakly immuno-positive against the anti-thyroxine (T₄) antibody. The number of thyroid follicles and the immunoreactivity later increased as the larvae grew. Thyroid hormones (TH) T₄ and triiodothyronine (T₃) were not detected in premetamorphic larvae by radioimmunoassays, but became detectable during metamorphosis. The maximum level of T₄ was seen in fish at the end of metamorphosis and in just-metamorphosed juveniles, whereas T₃ reached the highest level during metamorphosis and declined toward the end of metamorphosis. The results indicated that the thyroid gland first became active during metamorphosis in the development of eel.     

Supplemental effect of vitamin A in diet on the reproductive performance and egg quality of the Japanese flounder Paralichthys olivaceus (T & S)

This study investigated supplemental effects of vitamin A in diet on reproduction of Japanese flounder Paralichthys olivaceus (T & S). Broodstock were fed either a non‐supplemented diet (control diet; CD) or a vitamin A‐supplemented diet (supplemental diet; SD) 1 month before and during the spawning season. Both dietary groups began to spawn in mid‐January. At the beginning of spawning, no notable differences were found in spawning or egg quality between treatments. Egg production of the CD broodstock began to decrease from mid‐April and spawning stopped in late May, whereas the SD group continued to spawn until the end of June. Buoyant egg rate of the CD group was lower than that of the SD group throughout the spawning period. Hatching rate was not different between the CD and SD groups. The percentage of normal larvae in the CD group was significantly lower than in the SD group. It is concluded that a low vitamin A content in the diet has some negative effects on reproduction of flounder. The importance of supplementation of vitamin A in broodstock diet of the Japanese flounder is suggested.     

Reducing sea turtle by-catch in pelagic longline fisheries

Reducing by‐catch of sea turtles in pelagic longline fisheries, in concert with activities to reduce other anthropogenic sources of mortality, may contribute to the recovery of marine turtle populations. Here, we review research on strategies to reduce sea turtle by‐catch. Due to the state of management regimes in most longline fisheries, strategies to reduce turtle interactions must not only be effective but also must be commercially viable. Because most research has been initiated only recently, many results are not yet peer‐reviewed, published or readily accessible. Moreover, most experiments have small sample sizes and have been conducted over only a few seasons in a small number of fisheries; many study designs preclude drawing conclusions about the independent effect of single factors on turtle by‐catch and target catch rates; and few studies consider effects on other by‐catch species. In the US North Atlantic longline swordfish fishery, 4.9‐cm wide circle hooks with fish bait significantly reduced sea turtle by‐catch rates and the proportion of hard‐shell turtles that swallowed hooks vs. being hooked in the mouth compared to 4.0‐cm wide J hooks with squid bait without compromising commercial viability for some target species. But these large circle hooks might not be effective or economically viable in other longline fisheries. The effectiveness and commercial viability of a turtle avoidance strategy may be fishery‐specific, depending on the size and species of turtles and target fish and other differences between fleets. Testing of turtle avoidance methods in individual fleets may therefore be necessary. It is a priority to conduct trials in longline fleets that set gear shallow, those overlapping the most threatened turtle populations and fleets overlapping high densities of turtles such as those fishing near breeding colonies. In addition to trials using large 4.9‐cm wide circle hooks in place of smaller J and Japan tuna hooks, other fishing strategies are under assessment. These include: (i) using small circle hooks (≤ 4.6‐cm narrowest width) in place of smaller J and Japan tuna hooks; (ii) setting gear below turtle‐abundant depths; (iii) single hooking fish bait vs. multiple hook threading; (iv) reducing gear soak time and retrieval during daytime; and (v) avoiding by‐catch hotspots through fleet communication programmes and area and seasonal closures.     

Season- and Age-related Reproductive Changes Based on Fecal Androgen Concentrations in Male Koalas,Phascolarctos cinereus

The purposes of the present study were to clarify age- and season- related androgen patterns, and to compare the reproductive physiology between Japanese captive koala populations and Australian populations. To measure fecal androgens, feces were collected from male koalas (4.2 to 13.8 years of age) kept in Japanese zoos. Fecal androgens were extracted with methanol from the lyophilized samples and determined by enzyme immunoassay using 4-androstene-3,17-dione antibody. Fecal androgen concentration in male koalas increased after sexual maturation and remained relatively high until old age. In the survey with the Japanese zoo studbook of koalas, copulation (conception) month showed a pyramid shape with a peak in March to June (60.7%) in koalas born and reared in Japanese zoos and from July to April with the highest concentration in September to January (69.7%) in Australian institutes. Japanese zoo koala populations have a characteristic physiological cycle adapted to Japan''s seasonal changes. The suitable month of year for copulation or conception in Japan is diametrically opposed to that in Australia. Mean fecal androgen concentrations by month in the males born and reared in Japan indicated annual changes with the highest concentration in May and the lowest value in November. Fecal androgen analysis may be a noninvasive alternative tool to monitor circulating testosterone and may be helpful in understanding reproductive activity and physiology in male koalas.     

Proposal of screening method for intestinal mucus adhesive lactobacilli using the enzymatic activity of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH)

Adhesion tests are complex, time‐consuming and expensive, while the most important criterion for a probiotic lactobacilli is the ability to adhere to the human intestine. Thirty lactobacilli isolates from human intestinal tissues were measured for cell surface glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) activity using a microtiter plate screening method. GAPDH activities were detected in 21 out of 30 samples from 12 h cultures and in all samples from 18 h cultures. This suggests GAPDH is universally expressed on the bacterial cell surfaces from many lactobacilli. A statistically significant positive correlation was shown between GAPDH activity and adhesion using the BIACORE adhesion assay (P < 0.01). The new screening method using GAPDH enzymatic activity without an adhesion test may be possible due to the significant positive correlation of GAPDH activity with adhesion of lactobacilli derived from the human intestine.     

Determining the S-genotypes of several sweet cherry cultivars based on PCR-RFLP analysis

Summary

Based on the cDNA sequences encoding sweet cherry self-incompatibility associated ribonucleases (S-RNases), a PCR-based S-allele typing system for sweet cherry cultivars has been recently developed. Using this technique, we determined S-genotypes of the three newly released Japanese cvs Kouka-Nishiki, Beni-Sayaka and Beni-Shuho and one British cv Merton Glory that was classified as a Universal Donor, which is able to be used as a pollen donor for all cultivars in pollen incompatibility groups I to XIII. Furthermore, we also determined the partial sequences of the S-RNase genes of ‘Rainier’ (S¹S⁴)‘ and ‘Sato-Nishiki (S³S⁶)’,which leads to the development of a more reliable S-allele identification method of PCR-RFLP for sweet cherry cultivars. Total DNA isolated from leaves of the four cultivars along with those from ten cultivars with known S-genotypes were PCR amplified with two sets of primers that were designed from DNA sequences encoding the signal peptide (Pru-T2) and two conserved domains (Pru-C2 and Pru-C4R) of sweet cherry S-RNases. By comparing the size of PCR products on agarose gel, the 5-genotypes of ‘Kouka-Nishiki’, ‘Beni-Sayaka’, ‘Beni-Shuho’ and ‘Merton Glory’ were suggested to be S¹S³, S¹S⁶, S⁴S⁶, and S⁴S⁶, respectively. Two of these three S-genotypes (S¹S⁶ and S⁴S⁶) were found for the first time. DNA sequencing of PCR products from S-alleles of ‘Rainier’ and ‘Sato-Nishiki’ revealed that Ban II, Nru I, Apa LI and Ava I sites, respectively, were unique in the S¹-, S³-, S⁴- and S⁶- sequences flanked by Pru-T2 and Pru-C4R primers. RFLP analysis of the PCR products using these enzymes confirmed that S¹-, S³-, S⁴- and S⁶-alleles of the four cultivars contained the respective restriction enzyme recognition sites.     

Correlation between skeletal muscle fiber type and responses of a taste sensing system in various beef samples

The difference of muscle fiber type composition affects several parameters related to meat quality; however, the relationship between muscle fiber types and meat taste is unclear. To elucidate this relationship, we determined the taste of various beef samples using a taste sensor (INSENT SA402B) and analyzed its correlation with different muscle fiber type composition. We used 22 kinds of beef samples and measured nine tastes, including the relative and change of membrane potential caused by adsorption (CPA) values, using six sensors (GL1, CT0, CA0, AAE, C00, and AE1). The taste sensor analysis indicated positive value outputs for the relative C00, AAE, and GL1 values as well as for the CPA value of AAE, which corresponded to bitterness, umami, sweetness, and richness, respectively. We found significant positive correlations of the myosin heavy chain 1 (MyHC1) composition with umami taste, and with richness. This result suggests that high levels of slow MyHC1 can induce strong umami taste and richness in beef. We expect that our results will contribute to the elucidation of the relationship between muscle fiber types and meat palatability.     

Evaluation of the Subchronic Toxicity of Dietary Administered Equisetum arvense in F344 Rats

Equisetum arvense, commonly known as the field horsetail, has potential as a new functional food ingredient. However, little information is available on its side effects, and the general toxicity of Equisetum arvense has yet to be examined in detail. In the present study, we evaluated the influence of administration in diet at doses of 0, 0.3, 1 and 3% for 13 weeks in male and female F344 rats. No toxicity was detected with reference to clinical signs, body weight, urinalysis, hematology and serum biochemistry data and organ weights. Microscopic examination revealed no histopathological lesions associated with treatment. In conclusion, the no-observed-adverse-effect level (NOAEL) for Equisetum arvense was determined to be greater than 3% in both sexes of F344 rat (males and females: >1.79 g/kg BW/day and >1.85 g/kg BW/day, respectively) under the conditions of the present study.