Bovine leukemia virus (BLV) is a type C retrovirus infecting bovine B cells and causing enzootic bovine leukosis. Since it takes long periods to develop the disease, it is believed that BLV and host immune responses are closely related. In this review, the accumulated data showing close relationship between BLV and host immune responses are summarized in 4 sections. First, we discuss the role of cell-mediated immunity in protecting hosts from BLV infection. Second, several reports showing the relationship between the disease progression and the change of cytokine profiles are summarized. In the third section, we have focused on tumor necrosis factor α (TNFα) and its two types of receptors, and the possible involvement of TNF α in the BLV-induced leukemogenesis is discussed. The expression of TNF α has been shown to be regulated by major histocompatibility complex (MHC) haplotype. The resistance to BLV infection is supposed to be established by some innate factors, which are closely related to MHC haplotype. Finally, we propose that a breeding strategy based on the MHC haplotype could be a good approach to control BLV infection. This review includes some recent data from us and other groups.

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Differential maturation of three types of olfactory organs, the olfactory epithelium (OE), the vomeronasal organ (VNO) and the septal olfactory organ of Masera (MO), was examined immunohistochemically in embryonic and newborn rats by the use of anti-protein gene product 9.5 (PGP 9.5) serum. These olfactory organs were derived in common from the olfactory placode as neuroepithelia. In the OE, PGP 9.5-immunopositive olfactory cells first appeared at 13 days of gestation. The OE maturated completely, and showed the same cytological features as in the adult at 20 days of gestation. The MO first appeared as a dense mass of PGP 9.5-immunopositive sensory cells on the most ventrocaudal part of the nasal septum at 15 days of gestation and was evidently isolated from the OE by the decrease of immunopositive cells in the intercalated epithelium between the OE and the MO at 20 days of gestation. However, even at 7 days after birth, the MO did not complete its development and contained sensory cells aggregating in the mass. The VNO was separated from the nasal cavity at 13 days of gestation as a tubular structure of a neuroepithelium including PGP 9.5-immunopositive sensory cells. These cells gradually increased in number in the sensory epithelium of the VNO and extended their dendritic processes to the free surface at 7 days after birth. These findings clarified the differential maturation of these olfactory organs. That is, the OE completes its development before birth, while the MO and VNO after birth.

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The cycle of the seminiferous epithelium in the Java fruit bat, Pteropus vampyrus, and the Japanese lesser horseshoe bat, Rhinolophus cornutus, was investigated by light microscopy and the characteristics of spermiogenesis were compared between these two species. In the Java fruit bat, the cycle of the seminiferous epithelium was divided into 11 stages and developing spermatids were subdivided into 13 steps. While in the Japanese lesser horseshoe bat, the cycle of the seminiferous epithelium was divided into 10 stages and developing spermatids were subdivided into 13 steps. Excepting slight morphological differences, the characteristics of acrosomal formation in both species were almost similar with each other. In the Java fruit bat after stage VII, the acrosome gradually elongated, flattened and finally became scoop-like in shape. In the Japanese lesser horseshoe bat after stage VIII, the acrosome elongated, flattened and then slightly shortened. Before spermiation, the acrosome became long spatula-like in shape. The elongation and flattening of spermatids in these two species were similar to those in insectivores. The finding may reflect the fact that the order Chiroptera is phylogenetically close to the order Insectivora.

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The ICR-derived glomerulonephritis (ICGN) mouse is a novel inbred mouse strain with a hereditary nephrotic syndrome, considered to be a good model of human idiopathic nephrotic syndrome and develops proteinuria, hypoproteinemia and anemia. In the present study, we compared the cell kinetics in the kidneys of ICGN mice with age-matched ICR mice as normal controls. The proliferating cells were visualized by 5-bromo-2'-deoxyuridine labeling, and apoptotic cells were determined by terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end-labeling. Many proliferating epithelial cells of renal tubules, glomerular mesangial cells and tublointerstitial fibroblast-like cells were observed in the kidneys of ICGN mice, but no proliferating cells were seen in the kidneys of ICR mice. Apoptotic cells had round nuclei, and were observed only in the tubulointerstitium in the kidneys of ICGN mice but not in that of controls. The proliferation of renal tubular epithelial cells may represent a compensatory response, and that of mesangial and fibroblast-like cells may play a pathogenic role in nephrotic syndrome. Apoptosis in tubulointerstitial cells with round nuclei may have been erythropoietin-producing cells, and probably caused anemia.

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Naturally infected cases of swine mycobacteriosis were divided into two groups, localized infection (LI) and disseminated infection (DI). Lymphoproliferative response (LPR) was then examined to estimate their immunological states. Both control and LI groups showed strong response to Concanavalin A (Con A) and phytohemagglutinin (PHA) in the LPR, and lymphocytes recovered from the LI responded well to purified protein derived from M. avium (PPD). On the other hand, the DI group showed weak response to both Con A and PHA, despite their strong response to PPD stimulation. These data suggest that the low LPR to Con A and PHA observed in the DI groups was probably not due to the general unresponsiveness of T-cells.

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When female mice are mated, they form a memory to the pheromonal signal of their male partner. The neural mechanisms underlying this memory involve changes at the reciprocal dendrodendritic synapses between glutamatergic mitral cells and γ-aminobutyric acid (GABA)-ergic granule cells in the accessory olfactory bulb (AOB). Blockade of GABA_A receptors in the AOB leads to the formation of an olfactory memory. In an attempt to disrupt memory formation at mating, we used local infusions of the GABA_A receptor agonist muscimol into the AOB during the critical period for memory formation. Muscimol across a wide range of doses (1-1000 pmol) did not prevent memory formation. The resistance of this memory to GABA_A receptor activation may reflect the complexity of synaptic microcircuits in the AOB.

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Although mast cells contribute to host protective immunity against bacterial infections, the exact mechanism of their recruitment at the affected site has been unclear. Recently, we have reported that both mouse and human mast cells are capable of producing matrix metalloproteinase (MMP)-9, a matrix-degrading enzyme necessary for leukocyte transmigration. Here, we demonstrated that bacterial lipopolysaccharide (LPS) enhanced MMP-9 production of mouse bone marrow derived-cultured mast cells. This action of LPS was partially suppressed by the pretreatment of cultured mast cells with a protein kinase C (PKC) inhibitor, indicating the possible involvement of PKC signaling pathways in the production of MMP-9 by LPS. Thus, these suggest the upregulation of mast cell MMP-9 by bacterial components, thereby resulting in their migration at the affected site.

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An anomalous shunt between the bronchoesophageal artery and pulmonary artery was diagnosed in a 1-year- old, 3.5 kg female Miniature Dachshund by selective contrast angiography. A cardiac murmur had been observed in the dog during examination at another hospital. The machinery murmur was auscultated at the left side of the base of the heart. Although thoracic radiography revealed mild cardiomegaly, the characteristic findings of patent ductus arteriosus (PDA), including as aortic arch enlargement and pulmonary artery enlargement were not observed. Echocardiography demonstrated shunting of blood flow presumably from the arterial duct at the pulmonary artery carina. Based on the above findings the case was diagnosed as PDA. Angiocardiography was performed to confirm the diagnosis in preparation for surgical treatment, but later we confirmed that the shunt vessel was not PDA, but apparently a branch of the bronchoesophageal artery. The shunt vessel was branching in a complicated manner and shunted to the pulmonary artery.

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When lung fibroblast cell lines from LEC and WKAH rats were irradiated with ultraviolet B (UVB) and assayed for colony formation, LEC rat cells showed a higher sensitivity than did WKAH rat cells. The LEC rat cells were approximately 1.5-fold more sensitive to UVB radiation than were the WKAH rat cells in terms of D₃₇ values, which are the doses of UVB required to reduce cell survival to 37%. When the rat cells were irradiated with UVB in the presence of 0.5 M dimethyl sulfoxide (DMSO), which efficiently scavenges free radicals such as hydroxyl radicals, no significant difference was observed between the survival curves of either LEC or WKAH rat cells irradiated with UVB in the presence of 0.5 M DMSO and those irradiated with UVB in the absence of DMSO. Therefore, formation of free radicals may not be involved in cell death induced by UVB radiation. Flow cytometry showed that the percentage of apoptotic cells in the LEC rat cell population increased with post-incubation time after UVB radiation. The proportion of apoptotic cells in the UVB-irradiated LEC rat cell population increased as the dose of UVB was increased. In contrast, no significant proportion of apoptotic cells was observed in the UVB-irradiated WKAH rat cell population. These results showed a higher sensitivity in induction of apoptosis by UVB radiation in LEC rat cells than in WKAH rat cells.

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Reproductive organs of stained and mounted whole specimens of different types of Fasciola (F. hepatica, F. gigantica, and parthenogenetic diploid and triploid flukes) were observed to clarify the structure of their reproductive organs. The results are as follows; 1. Basic structure differences could not be identified. 2. The flukes without sperm, or those with an extremely small quantity in the seminal vesicle, are parthenogenetic Fasciola sp. 3. It was newly discovered that the surface of the cirrus is surrounded by many shallow gutters, and that spines form a line in the gutters. 4. The structure of the reproductive organ on the genus Fasciola are shown in detail in the figures.

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The antibody response to the recombinant protein, R32tet32, which contained the repetitive sequence (NANP)_n of Plasmodium falciparum CSP was determined in C57BL/6 mice during the course of nonlethal infection with Plasmodium yoelii 17X. Marked suppression of the IgG antibody response to R32tet32 occurred when mice were immunized at peak parasitemia (on day 16). In vitro antibody responses of spleen cells from acutely infected mice to R32tet32 were similarly suppressed. Stimulation of normal spleen cells cultured for 5 days with 100 ng/ml of R32tet32 gave an optimal IgG antibody response, but spleen cells from infected mice obtained at peak parasitemia failed to respond to a broad range of antigen concentrations. Cocultivation studies employing enriched lymphocyte populations from infected and uninfected C57BL/6 mice indicated that both T and B cells from infected mice were defective in their response to R32tet32. The response to the repetitive region was restored by the addition of recombinant mouse interleukin-2 (IL-2) at a dose of 50 U/ml to cultures of spleen cells from infected mice.

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Reticulocytes, known as late polychromatic erythrocytes, were induced in blood of chickens infected with Leucocytozoon caulleryi by bleeding when the second-generation merozoites were released into the blood from the second-generation schizonts. The second-generation merozoites preferentially invaded into reticulocytes and developed to stage II gametocytes. Enhanced development of stage II gametocytes to mature gametocytes was observed in the reticulocytes in vivo and in vitro in the bleeding group. Nevertheless, invasion of reticulocytes by the second-generation merozoites was not considered to be absolutely necessary for the development of gametocytes.

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Species-specific nested polymerase chain reaction (PCR) was used to detect the presence of possible canine ehrlichial agents (Ehrlichia canis, E. chaffeensis, E. ewingii, E. equi and E. platys) and monocytic ehrlichial agents found in Japan (E. muris and a recently discovered Ehrlichia species detected from Ixodes ovatus) in blood samples from dogs in Yamaguchi and Okinawa Prefecture, Japan. Partial sequence of E. platys was detected from 1 of 67 dogs (1.5%) tested from Yamaguchi Prefecture and 24 out of 87 (27.6%) in the subtropical Okinawa Prefecture. Dogs in Okinawa and Miyako Islands had a higher positive rate (69.2 and 45.0%, respectively) than Ishigaki Island (11.1%). Another dog in Yamaguchi Prefecture had a positive PCR reaction to the Ehrlichia sp. detected from I. ovatus. No other Ehrlichia were found in these samples.

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A case of wildlife trichinosis was found in a red fox (Vulpes vulpes japonica) captured at Rokkasho, Aomori Prefecture on November 27, 1998. Trichinella larvae were obtained from almost all of the muscle tissues except for the masseter. The highest number of larvae per gram of tissue was found in the muscles of the gluteal region and throat. The lowest number was found in the diaphragm and tongue. Trichina cysts within the muscle fibers had groups of fatty cells at the poles, and minimal tissue reaction was observed around the cyst. No calcification was found in the cyst. These morphological findings suggested that the considerable time had elapsed since the invasion. This is the first case of trichinosis in a red fox in Japan.

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Live virus vaccines for human use, 29 monovalent vaccines against measles, mumps, rubella or polio, eight polyvalent vaccines against measles-mumps-rubella and one bacterial polyvalent vaccine against Streptococcus pneumoniae, were tested by reverse transcriptase-nested PCR for the presence of petivirus or pestivirus RNA. Twenty-four samples were selected from European manufacturers, ten were from U.S.A. and four from Japan. Five (13.1%) out of 38 tested samples were positive for pestivirus RNA. Three vaccines (rubella and two measles) were from Europe and two (mumps and rubella) from Japan. The 5'-untranslated genomic region of the contaminant pestivirus RNA were amplified by reverse transcription-PCR and sequenced. Analyses based on primary nucleotide sequence homology and on secondary structures, characteristic to genotypes, revealed that the cDNA sequences belonged to bovine viral diarrhea virus (BVDV). A cDNA sequence, detected from one measles sample, belonged to BVDV-1b genotype. Pestiviral cDNA detected from the Japanese mumps and rubella vaccine samples, belonged to the BVDV genotypes 1a and 1c, respectively. Analysis on two cDNA sequences detected from measles and rubella vaccine samples from Europe showed their appurtenance to a new genotype, BVDV-1d. These findings indicate that contamination by animal pestivirus may occur in biological products for human use.

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To determine the process of formation of apical delta, a histological study on the permanent teeth was carried out in dogs. A litter of 7 clinically healthy beagle dogs and 33 adult dogs (4- to 15- year-old) of 12 breeds with periodontal disease were used for the experiments. Teeth extracted from 6-,7-,8- and 9-month-old beagles were sectioned and stained with HE solution. Tooth roots obtained from adult dogs with periodontal disease were ground. Each tooth was classified into the following root types under a light microscope: Type I (no apical delta = no apical closure), II (few apical delta), IIIA (low apical delta) and IIIB (high apical delta). In the 6-month-old beagles, more than half the tooth roots were classified as type I. In the 7-month-old beagles, type IIIB apical delta was the most predominant and types I, II and IIIA apical delta were occassionally seen. Apical closure and delta were observed in all beagles at 8 months of age histologically. In the 8- and 9-month-old beagles, all root apexes observed were type IIIB. Most of the 314 tooth roots extracted from 33 adult dogs were type IIIB, but a few were type IIIA.

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To establish a prediction table of parturition day the real-time B-mode ultrasonographic examinations were performed in the 8 pregnant Malteses and 10 Yorkshire terriers (total pups, 25 and 38 pups, respectively) from 18 days of gestation until the parturition. Ovulation was designated the first day of gestation (day 0). Extra fetal and fetal structures were measured from all conceptues. The parameters that exhibited the best correlation to parturition were used to compile a prediction table of parturition day. To testify the precision of the prediction table of parturition day, the 15 pregnant Malteses (48 pups) and 13 pregnant Yorkshire terriers (42 pups) with unknown mating time were examined using ultrasonography. Inner chorionic cavity diameter on days 18 to 37 and fetal head diameter on day 38 to parturition that showed the best correlation to gestational age were the most pertinent to the estimation of gestational age and the prediction of parturition day. The two parameters were used to compile a prediction table of parturition with averaged regression equations. In verificational examinations, with the exception of 1 Yorkshire terrier (3.6%) having 1 fetus, 18 of 28 bitches (64.3%) delivered exactly on the date predicted and 9 of 28 bitches (32.1%) delivered within 1 day of the date predicted. Therefore, the prediction table of parturition day seems to be a useful tool of the prediction of parturition day in practice.

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Four doses (equivalent to 4, 2, 1, and 0.5 liter water) of organic extracts from raw, treated and drinking waters sampled from seven different treatment plants in five cities in Korea were challenged to the Ames test using S. typhimurium strains TA98 and TA100 in the presence/absence of S9 mix. The mutagenicity was usually observed from chlorine-treated (28.6%) and drinking (42.9%) waters rather than raw (3.4%) waters. The strain TA98 (33.3%) was more sensitive to detect the mutagenicity of water samples than the strain TA100 (16.7%). However, the absence of S9 mix showed higher mutagenic activity of waters compared to the presence of S9 mix, corresponding to the detection of 42.9% and 7.1%, respectively. These results indicate that the bacterial mutagenicity of treated and drinking waters may be derived from chlorination in water treatment plants but that the mutagenicity in humans may be limited due to enzymatic metabolism.

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Classical swine fever virus (CSFV) strain WB82, isolated from a wild boar in 1982, induced a distinct cytopathic effect (CPE) in primary swine testicle cell culture and in most of the porcine cell lines. This strain of CSFV was found to be composed of two biotypes, cytopathogenic (cp) CSFV, as a minor population, and noncytopathogenic (noncp) CSFV, as a major population. The noncp CSFV (designated strain WB82/E⁺) was obtained by biological cloning, and it showed the exaltation of Newcastle disease virus phenomenon. In Northern blot analysis and RT-PCR assay, CSFV RNA with a subgenomic (sg) length was detected in addition to full-length viral RNA only in the cells in which a CPE had been revealed. These RNAs represent the genomes of typical defective interfering (DI) particles because of the strict dependence on a complementing helper virus and interference with replication of the helper virus. The sg RNA, which exhibits the genomes of the DI particles, lacked the nucleotides of the viral genomic region from N^pro to NS2 (4764 bases). When extracted sg RNA was transfected to the cells infected with the WB82/E⁺ strain, a distinct CPE was observed. Interestingly, the CPE was observed in cells infected with other heterologous noncp CSFV ALD and GPE^- strains by sg RNA transfection. The results suggested that these noncp CSFVs act as helper viruses for the replication of sg RNA (DI particles). It was also shown that the cytopathogenicity of strain WB82 is caused by apoptosis.

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The nucleotide sequence of the matrixprotein (M) gene of the lapinized rinderpest virus (RPV-L) was determined. The full-length cDNA of the RPV-L M gene is composed of 1460 base pairs and is supposed to contain an open reading frame of 1005 nucleotides encoding on M protein of 335 amino acids. The homology of the predicted amino acid among congeneric morbilliviruses such as RPV Kabete `O' strain (wild strain of RPV), RPV RBOK strain (vaccine strain of RPV for cattle), measles virus (MV), and canine distemper virus (CDV), is approximately 94%, 93%, 87% and 77%, respectively. In the present study, all coding regions of the RPV-L strain have been determined.

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