Aberrant development of mammary glands, but precocious expression of beta-casein in transgenic females ubiquitously expressing whey acidic protein transgene

It has been suggested that whey acidic protein (WAP) may function as a protease inhibitor. However, the actual function of WAP remains obscure. We investigated the histological development of the mammary glands of transgenic mice ubiquitously expressing WAP and CAG/WAP transgene. Ubiquitous expression of WAP induced aberrant development of the lobular alveoli of the mammary glands: mammary alveoli that were either aberrantly large or small in size increased in number in the developing mammary glands of these transgenic females during pregnancy and lactation. The expression of beta-casein was precociously induced in the mammary glands of the transgenic females during early pregnancy and accompanying this was a histological observation that abnormally developed lobular alveoli filled with milk proteins appeared in the mammary glands of transgenic females during early pregnancy. However, during lactation, the development of mammary glands was impaired in transgenic females. To investigate the possible paracrine action of WAP associated with mammary gland aberration, we transplanted the mammary tissue of CAG/EGFP transgenic females into the fat pad of virgin CAG/WAP transgenic females and initiated pregnancy by mating. The development of mammary tissue transplanted to the recipient was histologically examined on day 3 of lactation. The results revealed that the development of grafted mammary tissues was impaired in a manner similar to that of the mammary glands of CAG/WAP transgenic females, indicating that the inhibitory effect of WAP acts via a paracrine mechanism. In vitro experiments using HC11 cells with forced expression of exogenous WAP demonstrated the inhibitory function of WAP on proliferation of mammary epithelial cells.     

Role of actin microfilaments in canine distemper virus replication in vero cells

Several studies have indicated that viruses require a specific cytoskeletal structure for replication in host cells. In this study, we examined the role of actin fiber in the replication of canine distemper virus (CDV), belonging to the Morbillivirus genus of the family Paramyxoviridae. For this purpose, we used two actin depolymerizing agents, cytochalasin-D (C-D) and mycalolide-B (ML-B). In Vero cells, C-D disrupted actin fibers distributed in the cytosol, but peripheral actin fibers remained intact. On the other hand, ML-B completely disrupted the actin fibers distributed in both areas. Treatment of Vero cells with C-D or ML-B inhibited the replication of CDV. Double staining of CDV-infected Vero cells with antibody to N-protein and rhodamine-phalloidin revealed the presence of N-protein in mid-cytoplasm. However, the N-protein was specifically localized at the submembrane region in the presence of C-D, whereas it was clustered in the presence of ML-B. Viral mRNA levels of N- and H-proteins were rather increased by treatment with C-D or ML-B. The treatment with ML-B strongly inhibited N-protein expression, whereas C-D only slightly inhibited N-protein expression. These results suggest that actin microfilaments distributed in the cytoplasm and on the membrane region in host cells may have a different role in the process of CDV replication.     

The W‐ and Z‐linked EE0.6 sequences used for molecular sexing of captive Japanese crested ibis on Sado Island

The Japanese crested ibis Nipponia nippon is a critically threatened bird. Accurate sexing is necessary to perform effective management of captive breeding toward a national project for a tentative release of the Japanese crested ibis on Sado Island. A PCR‐based sexing method targeting a 0.6 kb EcoRI fragment (EE0.6) sequence on W chromosome with AWS03 and USP3 primers has been developed for the Japanese crested ibis. However, the primers were selected from the EE0.6 sequences from bird species other than the Japanese crested ibis. In this study, we determined the W‐ and Z‐linked EE0.6 sequences in the Japanese crested ibis, and clarified Japanese crested ibis sequence mismatch in the binding sites of the primers. Further, we found no polymorphism in the primer binding sites among five founder birds for the Sado captive Japanese crested ibis population. These findings validated the PCR‐based sexing method with the AWS03 and USP3 as accurate molecular sexing methods of captive Japanese crested ibis on the Sado Island. Additionally, we designed a primer set for a novel PCR‐based sexing, based on the EE0.6 sequences obtained in this study. This novel sexing method may be useful for future ecological research following the release of Japanese crested ibis on Sado Island. This is the first report to show the EE0.6 sequences in Japanese crested ibis.     

Antioxidant, anti-hyaluronidase and antifungal activities of Melaleuca leucadendron Linn. leaf oils

The present study examined the chemical composition, in vitro antioxidant, anti-hyaluronidase and antifungal activities of essential oils of Melaleuca leucadendron Linn. from Gundih-Central Java, Indonesia in different plant ages of 5, 10 and 15 years old. The Chemical composition of essential oils were analyzed by GC/MS. Twenty-six components were identified, of which 1,8-cineole (49.22–55.04 %), α-terpineol (8.79–10.70 %), d-limonene (5.58–6.39 %), and β-caryophyllene (5.03–7.64 %) were the main compounds in these oils. The antioxidant assay and anti-hyaluronidase assay showed that M. leucadendron leaf oils possess mild antioxidant activity with IC₅₀ between 7.21 and 9.23 mg/ml and anti-hyaluronidase activity with IC₅₀ between 1.94 and 3.03 mg/ml. The antifungal assay showed the effectiveness of these essential oils against Fomitopsis palustris (IC₅₀ 0.12–3.16 mg/ml), Trametes versicolor (IC₅₀ 0.01–0.06 mg/ml), Cladosporium cladosporioides (IC₅₀ 0.03–0.49 mg/ml), and Chaetomium globosum (IC₅₀ 0.06–0.15 mg/ml).     

Development of an air-injection press for preventing blowout of particleboard V: effects of board density and thickness on property of board manufactured from high-moisture particles

Particleboards with thickness of 10 mm and densities of 0.6, 0.7 and 0.8 g/cm³ were manufactured from high-moisture particles using urea–formaldehyde resin and the effectiveness of air injection was examined. The temperature in the 0.6 and 0.7 g/cm³ boards was lower with air injection than without during the initial to middle stages of pressing, while the temperature in the 0.8 g/cm³ board remained lower with air injection than without throughout the entire pressing process. Air injection reduced the pressing time required to manufacture the 0.6 and 0.7 g/cm³ boards and also increased the internal bond strength of boards of all densities. In the 0.6 and 0.7 g/cm³ boards, air injection reduced the modulus of rupture (MOR), while in the 0.8 g/cm³ boards, the MOR was similar between those manufactured by injecting and not injecting air. Air injection was also found to be effective for boards of high densities. The effectiveness of the air injection on thick boards was investigated by manufacturing 20-mm-thick boards of 0.7 g/cm³. Without air injection, it was not possible to manufacture the 20-mm-thick boards, even by extended hot pressing, but air injection allowed the boards to be manufactured by pressing for 16 min. Air injection was also shown to be effective for manufacturing thick boards.     

Performance of particleboard manufactured using air-injection press II: effects of board density and thickness on the performance of board manufactured from low-moisture particles

Particleboards of different densities (0.6, 0.7 and 0.8 g/cm³) and thicknesses (10 and 20 mm) were manufactured from low-moisture particles using an air-injection press. The effects of the air injection on preventing blowout of the boards of different densities and thicknesses were investigated by artificially creating blowout-prone conditions using metal frames. The effects of the air-injection pressure on the board performance were also investigated. 10-mm-thick boards of 0.8 g/cm³ pressed at 170 °C blew out when air was not injected, but were successfully manufactured by injecting air. 10-mm-thick boards at 150 °C showed constant internal bond (IB), regardless of density, but at 170 °C, IB was higher in boards of higher densities. This was likely due to accelerated hardening of the urea–formaldehyde resin at 170 than 150 °C. At both pressing temperatures, low air-injection pressure did not cause blowout and a reduction in board performance. Air injection also prevented the blowout of thick boards of 20 mm and enabled successful manufacture, showing its effectiveness. The IB of the 20-mm-thick board manufactured using the air-injection press exceeded that of 20-mm-thick board manufactured using an ordinary hot press.     

Dimensional stability and strength properties of particleboard produced by a closed-press system

The purpose of this study was to reveal the effects of various levels of mat-moisture content (m.m.c.) and the closed-press system for making single- or three-layer particleboard on the density profile, thickness swelling, specific moduli of elasticity (MOE) and rupture (MOR) and internal bond strength. Internal gas pressure was measured in an enclosed frame; and the larger the m.m.c., the higher the internal gas pressure became. When rising water vapor (steam) struck particles, it plasticized them and cured the adhesive, resulting in improved interparticle contact. The vertical density gradient in the three-layer board was larger than that in the single-layer board. As for thickness swelling by cold-water soaking, the single-layer boards were less affected than the three-layer boards and showed good dimensional stability with increased m.m.c. The open-system boards swelled more than the closed-system boards. The closed-system single-layer board made at high m.m.c. returned nearly to the prime thickness by air-drying after cold-water soaking. Specific MOE and MOR were larger at 15% or 10% m.m.c. than those at other m.m.c. Considerable reductions of specific MOR and MOE of the closed-system three-layer board were observed at 20% or 25% m.m.c.Part of this report was presented at the 45th annual meeting of the Japan Wood Research Society, Tokyo, April 1995 and at the 48th annual meeting of the Japan Wood Research Society, Shizuoka, April 1998     

Cytological and genetical studies on male sterility inCryptomeria japonica D. Don

Genetic male sterility is a useful trait in plant breeding, especially in angiosperm crops such as corn, onion and carrot. We found a male sterile sugi (Cryptomeria japonica D. Don) tree in Toyama, Japan. Pollen of sugi is one of the major causes of pollinosis in Japan. We carried out this research in an attempt to make clear the characteristics and inheritance of this male sterility. Microsporogenesis of the male sterile tree proceeded meiosis, however, the microspores collapsed after they were separated from pollen tetrads in locules, resulting in complete male sterility. Most likely, ethylene evolution was responsible for male sterility expression. Full seed setting in the male sterile tree indicated normal macrosporogenesis. Seeds obtained from crossing between male sterile and normal lines showed relatively high level of germination and their seedlings grew vigorously. The somatic chromosome numbers of 241 germinated seeds, derived from the male sterile tree, were mostly 22, euploid. These results indicated that male sterile tree was different from other similar previously reported trees with low pollen fertility, resulting from triploid or trisomics. Probably, male sterility in sugi is either nuclear genetic male sterility or cytoplasmic male sterility. The study was partially supported by Program for Promotion of Basic Research Actives for Innovative Biosciences.     

Proton Budgets of Forest Ecosystems on Volcanogenous Regosols in Hokkaido,Northern Japan

The proton budgets of deciduous and coniferous forest ecosystems on volcanogenous regosols in Hokkaido, northern Japan, were studied by measuring the biogeochemical fluxes (atmospheric deposition, canopy leaching, vegetation uptake and leaching from soil) at each site during a three year period. The proton budgets were developed for individual compartments of the ecosystem: vegetation canopy, organic and mineral soil layers. At both sites, atmospheric S deposition was the dominant proton source in the vegetation canopy. In organic horizons, dissociation of weak acids (bicarbonate and/or organic acids) and vegetation uptake of base cations were the dominant proton sources, and the net mineralization of base cations was the dominant proton sink. Atmospheric acid deposition was almost neutralized in the forest canopy and organic horizon. At both sites, weathering and/or ion exchange of base cations and protonation of weak acids (mainly bicarbonate) were the dominant proton sinks in the mineral soil. In both organic and mineral soil, internal proton sources (mainly vegetation uptake of base cations and dissociation of weak acids) exceeded external proton sources, indicating that acid deposition was not the main driving force of soil acidification in the studied forest ecosystems.