Effect of activation methods for bovine oocytes after intracytoplasmic injection

A principal nuclear transfer procedure is to inject a donor cell into the perivitelline space in an enucleated oocyte and then electric fusion is performed (cell fusion method). The effects of activation methods in reconstructed oocytes for the serum-starved somatic cell cloning procedure were investigated in this study by means of intracytoplasmic injection (i.c.i.). Bovine oocytes were enucleated at 18-22 h for in vitro maturation, and subsequently the nucleus of cumulus cell collected from Japanese Black Bulls (JBCC) after 5-7 days of starved culture was injected into the recipient cytoplast with a piezo-micromanipulator. At 1 h after i.c.i., reconstructed oocytes were stimulated with ethanol (ET) or calcium ionophore (CaI) as the first activation treatment, followed by cycloheximide (CHX) or 6-dimethylaminopurin (DMAP) treatment as the second activation. In the experiment on the first activation method, the proportion of reconstructed oocytes developing to the blastocyst stage was significantly (p<0.01) higher in the ET activation method than that with CaI (10.5% and 4.7%, respectively). And the experiment on the second activation method after ET treatment showed similar proportions of blastocyst development in both CHX and DMAP treatments (5.9% and 2.8%, respectively). The present results indicated that combined activation treatment with ET and CHX was efficient for reconstructed bovine oocytes by i.c.i.     

Clinical response of ovarian cysts in dairy cows after PRID treatment

We investigated the therapeutic effects of a progesterone releasing intravaginal device (PRID) on cystic ovarian disease (COD) and reproduction performance of cows. The possible influence of PRID on metabolic and/or health status was also examined. A total of 40 Holstein-Friesian cattle, with ovarian cystic structures, > or =2.5 cm in diameter, persisting for more than 7-14 days, without a corpus luteum (CL) were used for the study. PRID or placebos were inserted into the vagina for 12 days. Five animals lost the intravaginal device before removal and one was culled. Based on plasma progesterone concentration on the day of treatment, 20 (17 PRID and 3 placebos) of the remaining 34 cows had follicular cysts (progesterone < or =1 ng/ml) and 14 (10 PRID and 4 placebos) had luteal cysts (progesterone >1 ng/m l). Fourteen (82%) of the PRID-treated follicular cystic cows responded with formation of a CL within 14 days after treatment, and an overall conception rate of 53.8%. Likewise, 70% of the treated luteal cystic cows responded with CL formation and 71.4% conception rate. No significant differences were observed in hematocrit (Ht), white blood cell count and serum levels of glucose, blood urea nitrogen, aspartate aminotransferase, and alanine aminotransferase, between the day of PRID insertion and removal, in animals with follicular and luteal cysts. PRID treatment resulted in ovulation 2-4 days later and formation of a CL in cows that recovered.     

Percutaneous ultrasound-guided over-the-wire catheterization of the portal and hepatic vessels in cattle

A new method for catheterization of the portal and hepatic veins in cattle by means of the over-the-wire system was investigated to maintain more reliable long-term patency of catheters. Four cattle were used to evaluate the success rate, patency and safety of the procedure. The catheters, coated by urokinase were patent as long as they were in situ. In addition, the introducer was useful to prevent the catheter from being broken. No complications developed during the10 days after the procedure. Two cows were then euthanized. Post mortem findings were minimal. The results of the study reported here are promising, the benefits are significant and there is no apparent disadvantage to its use.     

Comparison of macrophage scavenger receptor-A knockout mice with wild type ones in the immune response against repeated infestation with Haemaphysalis longicornis

Using macrophage scavenger receptor-A knockout (SRKO) mice, we examined the role of macrophage class A scavenger receptors (MRS-A) on the immune response and acquisition of host resistance against repeated infestation with Haemaphysalis longicornis. Except for one batch of nymphs that infested one of the SRKO (SR-/-) mice and showed no appreciable reduction in body weight, all the other groups of nymphs manifested significant decrease in body weight. Both SR-/- and wild type (SR+/+) mice showed a sustained increase in anti-tick antibody titers, but SR+/+ mice showed significantly higher titers. The IFN-gamma assayed in SR-/- mouse immune sera was substantially less compared with that in SR+/+ mice. Immune sera from SR-/- and SR+/+ mice recognized the 51 and 44 kDa, and 44 kDa proteins, respectively, of the salivary gland antigen. The difference in the level of anti-tick resistance manifested by both groups of mice may be influenced by less efficient trapping and processing of tick antigens by macrophages in mice lacking for the macrophage scavenger receptors, and consequently affected the cascade of Th1 and Th2 responses. We have thus obtained valuable data that strongly infer the role of MSR-A in enhancing host defense against repeated infestation with H. longicornis.     

Assessment of wood surface roughness: comparison of tactile roughness and three-dimensional parameters derived using a robust Gaussian regression filter

Japanese oak and Japanese beech were sanded by hand with abrasive papers of varying grit number. Two three-dimensional parameters selected to characterize their surface roughness – one parameter for the distribution of roughness-profile peaks and the other for the relative area of the roughness-profile peaks above the threshold height – were compared against tactile roughness. The parameters were obtained from roughness profiles as determined by a robust Gaussian regression filter (RGRF) using seven cutoffs. The RGRF filtering process was adjusted specifically for the evaluation of wood surface roughness. Except for a cutoff wavelength of 0.25mm, the RGRF lent itself well to the determination of roughness profiles. No distortion of roughness profiles occurred around deep valleys, and there was a good correlation between the parameters and tactile roughness.Part of this study was presented at the 2002 Kansai Branch Office lecture meeting of the Japan Society for Precision Engineering, Kyotanabe, August 2002     

Surface deterioration of wood-flour polypropylene composites by weathering trials

The market for wood-fiber plastic composites (WPCs) is expanding rapidly in many countries including Japan, where WPCs are mainly used for exterior products. In such applications, WPCs undergo undesirable color change, chalking, and shrinkage and swelling, and accordingly there is a need to better understand the mechanisms responsible for the weathering of WPC and develop methods of improving their weathering resistance. In this study, weatherability of WPC was assessed by natural and accelerated weathering trials. Discoloration (whitening) of WPC during exposure was caused by degradation of both wood and plastic. Darker color pigments as additives improved the color stability of WPC; however, chalking on the surfaces still occurred. The color stability of WPC was improved by application of exterior coatings. Preweathering of WPC (before coatings were applied) increased the absorption of coatings by the WPC and had a positive effect on the color stability and prevented chalking of the composites. Parts of this study were presented at the IUFRO XXII World Congress Meeting, Brisbane, August 2005 and the IAWPS 2005, Yokohama, November 2005     

Transverse swelling behavior of hinoki (Chamaecyparis obtusa) revealed by the replica method

Transverse swelling and its anisotropy in hinoki (Chamaecyparis obtusa) in several kinds of organic liquids and in water were investigated by means the replica method. There was more cross-sectional swelling of cell walls and cell wall thickness in earlywood than in latewood. Marked swelling toward cell lumens was observed in wood swollen in liquids that had higher swelling potentials than water. This suggests that the swelling of cell walls in these liquids is much greater than the external swelling. Feret's diameters of the cell lumens were reduced by swelling in all the observed cases except in the tangential direction of earlywood, suggesting that cell walls swell to a much less extent in width than in thickness. Deformation of cell shapes caused by the tensile force from the latewood were observed in the earlywood and in the transitional region from earlywood to latewood. When swollen in water, transverse swelling anisotropy caused only by the swelling in cell wall thickness were calculated to be 1.2 for the whole region over an annual ring and 1.4 for the earlywood. These values could not account for the external swelling anisotropy of 2.1. Considering obvious deformations of cell shapes in the earlywood and in the transitional region, we conclude that the interaction between earlywood and latewood is one of the prime factors contributing to the transverse swelling anisotropy of coniferous wood.Part of this report was presented at the 48th Annual Meeting of the Japan Wood Research Society, Nagoya, April 1998     

The effects of within-species and between-species variation in wood density on the photodegradation depth profiles of sugi (Cryptomeria japonica) and hinoki (Chamaecyparis obtusa)

Low density wood is more rapidly eroded than denser wood when exposed to the weather, possibly because it is more susceptible to photodegradation. Fourier transform infrared microscopy was used to examine: (1) the depth of photodegradation in earlywood and latewood of sugi (Japanese cedar) and earlywood of hinoki (Japanese cypress) exposed for up to 1500 h to artificial sunlight emitted by a xenon lamp (375 W/m² within the 300 to 700 nm spectral range); and (2) the relationship between the density of wood tissues and depth of photodegradation. The depth of photodegradation varied between species (sugi and hinoki) as well as within a growth ring (sugi earlywood and latewood), and there was an inversely proportional relationship between depth of photodegradation and wood density. These findings may explain why low density earlywood is more rapidly eroded than latewood during weathering, and more generally, why there is an inverse relationship between the density of wood species and their rate of erosion during artificial and natural weathering. Part of this work was presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004     

Pseudodynamic tests and earthquake response analysis of timber structures II: two-level conventional wooden structures with plywood sheathed shear walls

Pseudodynamic (PSD) tests were conducted on two-level timber structures with plywood-sheathed shear walls, which each had an opening of different configuration, to study the effects of the mechanical properties of the first and second levels on the earthquake response of the structure. The specimens had two-level conventional post and beam frames that were 3 m wide, 3 m deep, and 6 m high with plywood sheathings nailed on one face of the structure. The first and second levels had different opening configurations of window, door, or slit. Lateral forces were applied at the top of the first and second levels, calculating step by step the next displacement based on the North-South (NS) components of the 1940 El Centro earthquake. The test results were compared with those of the time-history earthquake response analysis using the lumped mass model and hysteresis model presented in the companion article (part I). The experimental and simulated results showed that the simulation by means of the lumped mass time-history earthquake response analysis predicted quite well the response of the first level, but tended to underestimate the response of the second level, and that the PSD tests of an individual wall system with the mass supported by that particular wall generally show a conservative estimate of the response.