In vitro effect of recombinant human granulocyte colony-stimulating factor on canine neutrophil apoptosis

Apoptosis is essential in eliminating neutrophils (polymorphonuclear leukocytes: PMNs) in animals. The suppression of PMN apoptosis is believed to be beneficial in eradicating pathogens and is implicated in the pathogenesis of human inflammatory diseases. In the present study, canine PMNs were stimulated with recombinant human granulocyte colony-stimulating factor (rhG-CSF) to investigate the in vitro effect on the apoptosis of canine PMNs. Apoptotic cell rates were assessed by flow cytometry in relation to the ability of PMNs to produce reactive oxygen species (ROS). Canine PMN apoptosis was markedly suppressed by rhG-CSF treatment, in association with the retention of the PMN ability to produce ROS. The addition of cycloheximide abolished this suppression by rhG-CSF. Moreover, canine PMNs, which were stimulated by rhG-CSF, expressed high levels of anti-apoptotic mcl-1 gene mRNA, as quantified by real-time polymerase chain reaction method. The results suggest that PMNs, stimulated by G-CSF, could work effectively over a longer period to eliminate pathogens, and that the prolongation of the PMN life-span might occasionally aggravate tissue injuries in dogs. In addition, the suppression of PMN apoptosis seems to be mediated by the induction of anti-apoptotic mcl-1 gene expression.     

Effects of DDAVP administrated subcutaneously in dogs with aspirin-induced platelet dysfunction and hemostatic impairment due to chronic liver diseases

To evaluate the hemostatic effects of desmopressin (DDAVP) in dogs with aspirin-induced platelet dysfunction and hemostatic impairment in chronic liver diseases, 3 microg/kg DDAVP was administrated subcutaneously. In aspirin-induced platelet dysfunction dogs (n=5), prolonged BMBT (buccal mucosal bleeding time) was shortened significantly after DDAVP injection (2.2 +/- 1.2 min, P<0.05). In dogs with chronic liver diseases (n=4), activated partial thromboplastin time (APTT) tended to shorten by 0.9 to 3.0 sec, and prolonged BMBT was shortened in two cases for 4.2 and 1.7 min after DDAVP injection. Therefore, the present results indicated that DDAVP shortened the prolonged BMBT in dogs with aspirin-induced platelet dysfunction and chronic liver disease. DDAVP might be helpful in hemostasis under invasive procedures such as biopsy or surgery for dogs with hemostatic impairment.     

An enzyme-linked immunosorbent assay with the recombinant merozoite protein as antigen for detection of antibodies to Eimeria necatrix

A cDNA library was constructed with Eimeria necatrix merozoite mRNA and immunologically screened by chicken sera against this parasite. One of the positive clones containing an insert of 879 nucleotides, pNP19, showed similarity to part of a published gene expressed in E. tenella merozoite by the homology search system. The inserted DNA was subcloned into baculovirus, and a 35-kD protein was expressed, purified, and used for the antigen in enzyme-linked immunosorbent assay (ELISA). Antibodies from the chickens vaccinated with the E. necatrix attenuated strain, Nn-P125, were detected from 14 days after vaccination by ELISA. The mean absorbance increased rapidly to a peak around 21 days after vaccination; thereafter, it began to decline. Even though some of the vaccinated chickens showed very low levels of antibody response to the recombinant protein 56 days after vaccination, they were protected against challenge with virulent strain of E. necatrix. The mean absorbances in sera from both vaccinated and nonvaccinated chickens highly increased 14 days after challenge. On the other hand, the antibody was not detected in ELISA when chickens were exposed to other Eimeria species such as E. tenella, E. acervulina, and E. maxima. These results demonstrate that this recombinant protein is suitable for detecting the specific antibody in chickens infected with both attenuated and virulent strains of E. necatrix.     

Detection and identification of the Candida species by 25S ribosomal DNA analysis in the urine of candidal cystitis

Candida species in clinical urine samples were identified directly by the newly developed method of PCR analysis on 25S ribosomal DNA (rDNA). Two dogs were referred to the Animal Medical Center, Nihon University School of Veterinary Medicine, Fujisawa, Kanagawa, Japan for the examination of chronic cystitis. Microscopic examination of urine samples from these dogs revealed yeast cells. Urine culture on Sabouraud's dextrose agar at 27 degrees C for 5 days produced white to cream colored colonies. The isolates were identifical to Candida albicans and C. parapsilosis by mycological examination, respectively. The nucleotide sequences of 25S ribosomal DNA from these urine isolates showed 99% similarity to those of a reference strain of Candida albicans or C. parapsilosis. The nucleotide sequences of 25S rDNA obtained directly from urine samples were also identical to C. albicans and C. parapsilosis, respectively. Confirming the results on the isolates cultured from the same urine samples. This PCR analysis method could be available for the direct identification of Candida species in urine samples within 2 days.     

Contribution of active defense responses in the limitation of fungal spread in the sapwood of living sugi (Cryptomeria japonica) tree

The participation of active defense responses in the limitation of fungal spread in the sapwood of living sugi tree was determined. Isolates ofGuignardia cryptomeriae were inoculated to living trees, fresh logs, frozen and thawed logs and autoclaved logs, and the spread of wood discoloration and fungal hyphae was assessed. In autoclaved logs and freeze-treated logs, wood discoloration and reaction zone formation were not observed, and fungal hyphae of both of virulent and avirulent isolates grew rapidly in the wood. In fresh logs, wood discoloration and the formation of a pale reaction zone and transition zone were observed. Virulent isolates spread rapidly, but the spread of avirulent isolates was inhibited. In living trees, wood discoloration was observed and reaction zones were formed at lesion margins. These results indicated that constitutive defense mechanisms had no effect on the spread of both of virulent and avirulent isolates, and that active defense responses were important in the limitation of spread of fungal hyphae, hence in the limitation of wood discoloration. Further, it appeared that avirulent, but not virulent, races ofG. cryptomeriae were easily inhibited even by weak host responses. This study was presented at 105th and 106th Annual Meeting of the Japanese Forestry Society.     

Acoustoelastic effect of wood II: Effect of compressive stress on the velocity of ultrasonic longitudinal waves parallel to the transverse direction of the wood

The changes in the velocity of ultrasonic waves propagating in wood parallel to the direction of applied stress are discussed. The ultrasonic mode was longitudinal waves traveling along the direction of applied stress with the compressive load applied parallel to the transverse direction of the wood. The ultrasonic velocities were measured by the sing-around method. The experimental results indicated the existence of an acoustoelastic phenomenon in the transverse direction of the wood. The percent change in the ultrasonic velocity was given as a function of the applied stress. The change in the velocity depended on the species and structural direction of the wood. That is, in the radial direction of hardwood, the ultrasonic velocity increased with increases in compressive stress at the initial stress level of less than 2MPa; it then gradually decreased with increases in stress. A change in velocity from an increase to a decrease was considered a unique phenomenon for wood. In contrast, in the radial direction of softwood and the tangential direction of hardwood, the ultrasonic velocity decreased with increases in stress from the beginning of loading. This phenomenon is also generally observed in metallic materials. The relations between velocity and stress at the initial stress level and between velocity and strain in the range of large deformation are represented by essentially straight lines. The acoustoelastic constants of wood were obtained from these relations at the initial stress level. The absolute values of the constants in the transverse direction of wood were larger than those for metals and were larger than those for the longitudinal direction of wood reported in our previous paper.This research was presented at the 1st Meeting of the Research Society of the Acoustoelastic Measurements in the Japan Society of Non-Destructive Inspection at Osaka, October 1996 and at the 47th Annual Meeting of Japan Wood Research Society at Kochi, April 1997     

First description and collection records of juvenile slime flounder <Emphasis Type="Italic">Microstomus achne</Emphasis> from the eastern and western coastal waters off Honshu Island,Japan

Morphological characteristics and body pigmentations of juvenile slime flounder Microstomus achne were described from one specimen [27.8 mm total length (TL)] collected off Niigata Prefecture from the Sea of Japan (35 m depth) in May 1993 and from three specimens (30.1, 36.2, and 40.9 mm TL) collected off Fukushima Prefecture on the Pacific coast of northern Japan (15 and 50 m depths) during April–May 2008 and 2009. These age-0 juveniles had a characteristically slenderly oval-shaped body and slightly protruded front nostrils. Sparsely distributed melanophores and xanthophores were clearly visible throughout the ocular side of one specimen, which was judged as a newly settled juvenile, whereas several spots aggregated by small melanophores were found in the others with elongated pectoral fins and small cycloid scales. Melanophores were only slightly detectable on the blind side. Comparisons of the depth distributions of slime flounder juveniles and sympatrically collected age-0 pleuronectids (Pseudopleuronectes herzensteini, Pseudopleuronectes yokohamae, Platichthys bicoloratus, and Dexistes rikuzenius) suggested that slime flounder can settle on broad nursery areas of coastal and offshore pleuronectid flatfishes, probably depending on local oceanic conditions during the protracted pelagic phase.     

Effect of the lateral growth rate on wood properties in fast-growing hardwood species

We investigated the feasibility of using several fast-growing tropical or subtropical hardwood species for timber production by measuring key wood qualities in relationship to the high rates of lateral growth. The trees tested were sampled from even-aged plantations of Acacia mangium, A. auriculiformis, hybrid Acacia (A. mangium × A. auriculiformis), Eucalyptus grandis, E. globulus, and Paraserianthes falcataria (Solomon and Java origin) that had already reached commercial harvesting age. The released strain of the surface growth stress (RS), xylem density (XD), microfibril angle (MFA), and fiber length (FL) were measured at the outermost part of the xylem at breast height in each tree. Results were then compared to the lateral growth rate (radius/age) at breast height, which provides a relative indicator of the amount of tree growth per year. Our findings indicated that RS was constant, regardless of lateral growth rate in each species. Similar results were observed for XD, MFA, and FL, with a few exceptions, suggesting that high growth rates do not intrinsically affect the wood properties of fast-growing tropical or subtropical species that have reached harvesting age. However, special attention must be paid to patterns of xylem maturation when developing plantations of such species.     

Acinic cell carcinoma in an African pygmy hedgehog (Atelerix albiventris)

Abstract: A male African pygmy hedgehog ( Atelerix albiventris ), estimated to be 3 years old, presented with exophthalmos and fixed abduction of the right eye. Radiographic examination revealed a retrobulbar tumor in the right orbital cavity. The mass was surgically resected but recurred 3 months later and the hedgehog died. There was no gross or microscopic evidence of salivary or lacrimal gland involvement of the tumor at surgery or at necropsy. The histopathologic, immunohistochemical, and ultrastructural findings were those of acinic cell carcinoma, the origin of which was unknown. This is the first known case of acinic cell carcinoma in an African hedgehog.