Hypothermic storage of porcine zygotes in serum supplemented with chlorogenic acid

The current study was conducted to investigate the effects of 100% foetal bovine serum (FBS) and 100% porcine follicular fluid (pFF) as a storage medium on the developmental competence of porcine zygotes stored at 25°C for 24 hr. Moreover, we evaluated the additive effects of chlorogenic acid (CGA) in the storage medium. When in vitro‐produced zygotes were stored at 25°C for 24 hr in tubes containing either tissue culture medium (TCM) 199 supplemented with 1 mg/ml bovine serum albumin (BSA), 100% of FBS or 100% of pFF, the rate of blastocyst formation was significantly higher in 100% of FBS than in BSA‐containing TCM 199. When the effects of CGA supplementation in 100% of FBS on the development of zygotes stored at 25°C for 24 hr was evaluated, more zygotes stored with 50 µM CGA developed to blastocysts compared with the other concentrations of CGA. When the formation date and quality of blastocysts derived from zygotes stored in 100% of FBS supplemented with 50 µM CGA were investigated, the highest ratio of blastocysts formation in the storage group appeared 1 day later than in the non‐stored control group. However, a higher proportion of blastocysts with apoptotic nuclei was observed in the stored group as compared to the non‐stored group. In conclusion, 100% of FBS is available for a short storage medium of porcine zygotes. The supplementation of 50 µM CGA into the storage medium improves the rates of blastocyst formation of zygotes after storage, but the quality of embryos from the stored zygotes remains to be improved.     

Expression of anthocyanin biosynthetic genes in Malus sylvestris L. ‘Mutsu’ non-red apples

Summary

The expression of five anthocyanin biosynthetic genes of chalcone synthase (MdCHS). flavanane 3-hydroxylase (MdF3H). dihydroflavonol 4-reducase (pDFR), anthocyanidin synthase (MdANS) and UDP glucose-flavonoid 3-O-glucosyltransferase (pUFGluT) was investigated during fruit development in the skin of non-red ‘Mutsu’ apples (Malus sylvestris (L) Mill. van domestica (Borkh) Mansf.). MdCHS, MdF3H and pDFR appeared in the skin of non-bagged fruit from 81 d after full bloom (DAFB) to 167 DAFB, but MdANS and pUFGluT did not appear. When the fruit was bagged during fruit development and unbagged before the maturation, anthocyanin was produced in the exposed portion but not in the shaded portion. All five genes including MdANS and pUFGluT were detected in the exposed portion at 165 DAFB. In the shaded portion, MdANS was detected, but pUFGluT was not. The skin colour of the shaded portion after unbagging in the bagged fruit was more yellow than that in non-bagged fruit. This result suggests that the yellowing of the skin may relate to the expression of MdANS. All five anthocyanin biosynthetic genes were detected at 20 DAFB despite the absence of anthocyanin. The roles of genes in the anthocyanin biosynthetic pathway may differ from one development stage to the next.     

Blood pressure characteristics of female spontaneously diabetic Torii-Lepr(fa) rats

Blood pressure in female SDT-fa/fa rats was periodically investigated at ages 8, 16, and 24 weeks. Furthermore, an insulin therapy was performed for 5 weeks in the female rats at age 11 weeks, and the change of blood pressure was examined. In addition to obesity, hyperglycemia, hyperinsulinemia, and hyperlipidemia, hyperleptinemia and increased urinary angiotensinogen level were observed during the experimental period. Blood pressure was elevated at ages 8 and 16 weeks, but that at 24 weeks was comparable to that in Sprague-Dawley (SD) rats. Heart rate was decreased from age 8 to 24 weeks. Insulin therapy induced good glycemic control and improvement of hyperlipidemia, but the blood pressure was not reduced. Blood pressure in female SDT-fa/fa rats was elevated temporarily. The blood pressure was not decreased by insulin treatment. SDT-fa/fa rat is a useful model to investigate the relation between diabetes mellitus and hypertension.     

Reversible control of hydrogenation of a single molecule

Low-temperature scanning tunneling microscopy was used to selectively break the N-H bond of a methylaminocarbyne (CNHCH3) molecule on a Pt(111) surface at 4.7 kelvin, leaving the C-H bonds intact, to form an adsorbed methylisocyanide molecule (CNCH3). The methylisocyanide product was identified through comparison of its vibrational spectrum with that of directly adsorbed methylisocyanide as measured with inelastic electron tunneling spectroscopy. The CNHCH3 could be regenerated in situ by exposure to hydrogen at room temperature. The combination of tip-induced dehydrogenation with thermodynamically driven hydrogenation allows a completely reversible chemical cycle to be established at the single-molecule level in this system. By tailoring the pulse conditions, irreversible dissociation entailing cleavage of both the C-H and N-H bonds can also be demonstrated.     

Challenges to controlling rabies in skunk populations using oral rabies vaccination: A review

Controlling rabies in skunk populations is an important public health concern in many parts of the United States due to the potential for skunk rabies outbreaks in urban centres and the possible role for skunks in raccoon rabies variant circulation. Oral rabies vaccination (ORV) programmes have supported wildlife rabies control efforts globally but using ORV to control rabies in skunk populations has proven more challenging than with other target species, like foxes, coyotes and raccoons. A review of published studies found that some ORV constructs are immunogenic in skunks and protect against virulent rabies virus challenges, especially when delivered by direct installation into the oral cavity. However, in field ORV programmes using currently available vaccine‐bait formats and distribution methods targeting other rabies reservoir species, skunks often fail to seroconvert. Field effectiveness of ORV in skunks appears to be limited by poor bait uptake or inadequate ingestion of vaccine rather than from poor vaccine efficacy. Observations of captive skunks revealed vaccine spillage when handling and biting into baits such that modification of bait formats might improve field effectiveness. In addition, a dose–response relationship between bait distribution density and post‐baiting seroconversion among skunks was observed across the limited number of field studies. Additional research is needed to identify opportunities to modify ORV baits and distribution strategies to improve the viability of ORV as a rabies control strategy in skunks.     

Drug resistance mechanism of the fish-pathogenic bacterium Lactococcus garvieae

The minimum inhibitory concentrations (MICs) of 15 chemotherapeutic agents were tested against 146 Lactococcus garvieae strains isolated from 1999 to 2006 in Japan. The agents used included chloramphenicol, ciprofloxacin, erythromycin (EM), enoxacin, fleroxacin, florfenicol, kanamycin, lincomycin (LCM), norfloxacin, oxolinic acid, orbifloxacin, ofloxacin, benzylpenicillin, streptomycin and tetracycline (TC). Of the tested strains, 46 showed high levels of resistance to EM, LCM and TC. Twelve of these strains were detected to be carrying transferable R-plasmids using a conjugation experiment and, using Southern hybridization, were shown to have the same structure as the R-plasmid. The remaining 34 resistant strains had a similar DNA structure to that of the R-plasmid as confirmed by polymerase chain reaction (PCR) using primers designed from sites in the transferable R-plasmid. The EM and TC resistance genes were classified into the ermB and tetS groups using PCR. We also detected gyrA and/or parC mutants that are highly resistant to old and new generation quinolones. This study revealed that transferable R-plasmids encoding EM, LCM and TC are widely distributed and are conserved regardless of the area and/or time of collection.     

Study of sugi (Cryptomeria japonica D. Don) with male-sterility gene using CAPS markers

The frequency of open-pollinated male-sterile seedlings from a male-sterile mother tree in Cryptomeria japonica D. Don was determined in 765 open-pollinated seedlings. Pollen was not formed by 29 out of the 765 seedlings. To find pollen donor candidates for 29 male-sterile seedlings, a male-sterile tree was crossed with five individuals in the neighborhood of the mother tree. Analysis of microspores in male flowers of F₁ seedlings revealed that one of the five individuals had a heterozygous male-sterility gene. Paternity tests were conducted on the 29 male-sterile seedlings using eight kinds of CAPS markers in C. japonica. The pollen parents of 29 male-sterile seedlings were not only a tree in the neighborhood of the mother tree, but also other trees. Since it was found that the pollen parents of four of the male-sterile seedlings were not in the study plot, it was inferred that C. japonica trees that possess the male-sterility gene grow in other places and may be widely distributed.     

Facing the Challenge of Social Forestry in Japan: The Case of Reviving Harmonious Coexistence Between Forest and People in Okayama Prefecture

Social forestry as a development strategy has evolved since the 1970s, especially in the tropics, to address forest degradation and promote local community development amidst the burgeoning population in these areas. As a practice, however, social forestry has been in place since ancient times in many parts of the world, including Japanese forest communities. Forest-people relationships in Japan drastically changed through massive afforestation programs after the energy source change and with the industrialization of the forest sector in 1950s. The majority of the planted forests are underutilized today and forest communities are marginalized due to the decline of forestry operations, depopulation, and changes in people’s values. Some communities address this concern by inviting potential urban migrants who may be interested in settling in rural areas. Using the case of the Nishiawakura Village in Okayama Prefecture, this paper explores the recent challenges confronting social forestry in Japan. It is found that underutilization of forest resources can be a cause of serious environmental degradation and marginalization of forest communities, and that Nishiawakura’s journey to renew forest management in partnership with migrants is a process of revisiting and creating the forest-people relationship. This study advances two related arguments, namely (1) the interaction of the local people and the migrants brings new perspectives to forest management, and (2) in a community facing depopulation and underutilization of forest resources, social forestry can be an effective approach to rediscover traditional forest management in a new form and revitalize forests and local communities.