Isolation and characterization of alpha 1-acid glycoprotein from horses, and its evaluation as an acute-phase reactive protein in horses.

Equine alpha 1-acid glycoprotein (alpha 1AG) was isolated from equine serum by successive ammonium precipitation, anion- and cation-exchange chromatographies, and gel filtration. Purified equine alpha 1AG had a molecular weight of 46,000 +/- 1,000, and contained 31.4% carbohydrate. Gel isoelectric focusing revealed an isoelectric point range of 2.8 to 3.7. With immunoelectrophoresis, it was found that alpha 1AG migrated to the alpha 1-globulin region. Single radial immunodiffusion was used for quantitative measurement of alpha 1AG in equine serum. In clinically normal foals, serum alpha 1AG was undetectable (less than or equal to 20 micrograms/ml) in less than or equal to 7-day-old foals, but was detected by 14 days. The alpha 1AG concentration (mean +/- SD) increased to reach mean adult values of 99.23 +/- 26.90 micrograms/ml by 1 year of age. The alpha 1AG concentration in pregnant mares decreased at 2 to 3 months before parturition, then gradually increased until 1 day after parturition, when a brief decrease was observed. The concentration increased again at 2 weeks after foaling, then a decrease was observed, after which the alpha 1AG concentration increased again by 2 to 4 months after parturition. The concentration of serum alpha 1AG quickly rose to peak values 2 to 3 days after castration and jejunojejunostomy in adult horses, returning to baseline values by 14 to 28 days after surgery. The alpha 1AG was concluded to be an acute-phase reactive protein in horses.     

Zoonotic risk of Toxocara canis infection through consumption of pig or poultry viscera

The potential zoonotic risk of Toxocara canis infections from consumption of swine or poultry viscera containing larvae was assessed using a pig model. Two groups of six pigs were fed either fresh swine viscera (group FS) or poultry viscera (FP) containing around 3500 Toxocara larvae. Another two groups of six pigs were fed swine viscera (PS) or poultry viscera (PP) preserved at 4 degrees C for 1 week. All pigs were necropsied 14 days after the exposure. Liver white spots were counted and T. canis specific IgG antibodies were measured by ELISA. Larval burdens were assessed in the mesenteric lymph nodes, liver, lungs, brain, tongue, and eyes. All recipient pigs exhibited several white spots on the liver surface and detectable antibody levels. Larvae were recovered predominantly from the lungs, but also from the mesenteric lymph nodes and the liver, a few larvae were found in the brain and tongue of the pigs. Two larvae were found in the eyes of two pigs in group FS. Mean percentages of total larval recoveries in groups FS, FP, PS, and PP were 75.3, 63.6, 42.6, and 18.8%, respectively. Significantly higher numbers of larvae were recovered from pigs given swine viscera than pigs given poultry viscera. The preservation at 4 degrees C for 1 week caused a significant reduction in the larval infectivity overall, nevertheless, the recoveries remained substantial. The fact that larvae migrating in swine or poultry organs and tissues have high infectivity in pigs even after preservation at 4 degrees C for 1 week, suggests that human infection with T. canis might easily occur following consumption of raw or undercooked dishes, either fresh or refrigerated, prepared from swine or poultry organs and tissues harbouring T. canis larvae.     

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.     

Sporadic cases and an outbreak of leptospirosis probably associated with recreational activities in rivers in the northern part of Okinawa Main Island

In the summer of 2003, sporadic cases and an outbreak of human leptospirosis probably related to recreation in rivers occurred in the northern part of Okinawa Main Island. Sixteen of 22 suspected cases were definitely diagnosed as leptospirosis by serological test or isolation. The infective leptospiral serovar in 14 cases was presumed to be Hebdomadis. Transmission was thought to occur by exposure to river water that was contaminated by the urine of infected animals. The findings indicate that recreation in rivers in this area is a significant risk factor for infection with leptospires.     

Morphometry on lancet flukes found in Japanese sika deer (Cervus nippon centralis) captured in Iwate Prefecture, Japan

Thirty-six flukes were collected from the livers of wild deer (Cervus nippon centralis) captured in Iwate Prefecture, Japan, and were served for morphometry. The length and/or the width of the body, suckers, testes, ovary, vitelline glands, cirrus pouch and eggs in the uterus of the flukes were measured. The distance between anterior end of the body and position of the maximal body-width or upper end of the testes were also determined. A remarked morphological characteristic was that the right and left testes did not lie tandem but lined bilaterally. Also the position of the maximal body-width did not always locate in the posterior part of the body of the fluke. The property was in accordance with those for Dicrocoelium chinensis.     

Design of hammerhead ribozymes that cleave murine Sry mRNA in vitro and in vivo

As the first step in investigating the possiblity of applying ribozyme technology to artificial control of the sex ratios at birth in farm animals, where the demand for females exceeds that for males, we designed a hammerhead ribozyme (HHRz) and 2 tRNA(val)-hammerhead ribozyme complexes (tRNARz3 and tRNARz4), and examined their effects upon murine Sry mRNA in vitro and in cells. We demonstrated that HHRz and tRNARz3 could effectively cleave the target Sry mRNA in vitro. For the purpose of experiments in vivo, HHRz was cloned into the highly efficient pUC-CAGGS mammalian expression vector (pCAG/HHRz), and the tRNA ribozyme complexes were cloned into the pol III promoter-driven pPUR-KE vector (pPUR/tRNARz3 and pPUR/tRNARz4); the ribozyme vectors were co-transfected with the target vector (pCAG/Sry). A suppressive action (up to approx. 60%) was confirmed for pCAG/HHRz and pPUR/tRNARz3 upon the transiently expressed exogenously introduced Sry in M15 cultured cells.     

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.