Virulence of infectious pancreatic necrosis virus is associated with the larger RNA segment (RNA segment A)

Abstract. Infectious pancreatic necrosis virus (IPNV) contains two segments of double- stranded RNA (the larger RNA segment, A; and a smaller RNA segment, B). The IPNV- Buhl isolate belongs to the VR299 serotype, and eel virus European (EVE) is serologically the same as the Ab serotype. IPNV-Buhl is virulent to rainbow trout fry, Oncorhynchus mykiss (Walbaum), whereas EVE is avirulent. To determine the genetic basis for the difference of the virulence, intertypic reassortants were made between IPNV-Buhl and EVE. The reassortants with the genotypes of P/E (segment A of IPNV-Buhl and segment B of EVE) and E/P (segment A of EVE and segment B of IPNV-Buhl) were studied for their pathogenicity to rainbow trout fry. The P/E reassortant was highly virulent, similar to the IPNV-Buhl parent (P/P), while the E/P reassortant was avirulent, as was the EVE parent (E/E). A high level of viral replication was observed in almost all organs of the fry infected with the P/E reassortant and the IPNV-Buhl parent. In contrast, viral replication was restricted to the epidermal cells of the fry infected with the E/P reassortant and the EVE parent. The results demonstrate that the larger RNA segment (RNA segment A) of IPNV is related to cell tropism in vivo and is responsible for virulence in rainbow trout fry.     

Coincidental detection of genomes of porcine parvoviruses and porcine circovirus type 2 infecting pigs in Japan

The infection status of 15 viruses in 120 pigs aged about 6 months was investigated based on tonsil specimens collected from a slaughterhouse. Only 5 species of porcine parvoviruses and porcine circovirus type 2 (PCV2) were detected at high frequencies; 67% for porcine parvovirus (PPV) (PPV-Kr or -NADL2 as the new abbreviation), 58% for PPV2 (CnP-PARV4), 39% for PPV3 (P-PARV4), 33% for PPV4 (PPV4), 55% for PBo-likeV (PBoV7) and 80% for PCV2. A phylogenetic analysis of PPV3 suggested that Japanese PPV3s showed a slight variation, and possibly, there were farms harboring homogeneous or heterogeneous PPV3s. Statistical analyses indicated that the detection of PCV2 was significantly coincidental with each detection of PPV, PPV2 and PPV3, and PPV and PPV4 were also coincidentally detected. The concurrent infection with PCV2 and porcine parvoviruses in the subclinically infected pigs may resemble the infection status of pigs with the clinical manifestations of porcine circovirus associated disease which occurs in 3–5 months old pigs and is thought to be primarily caused by the PCV2 infection.     

Potential role of hCG in apoptosis of human luteinized granulosa cells

The corpus luteum (CL) forms after ovulation and acts as a temporary endocrine gland that produces progesterone (P4), a hormone that is essential for implantation and maintenance of pregnancy in mammals. In pregnant women, human chorionic gonadotropin (hCG) secreted by the conceptus prevents luteolysis. hCG also increases the survival of cultured human luteinized granulosa cells (hLGCs). To clarify the maintenance mechanism of the human CL, we investigated the effects of hCG and P4 receptor antagonists, onapristone (OP) and RU486, on the viability of hLGCs. With the patients’ consent, hLGCs were isolated from follicular aspirates for in vitro fertilization. The cells were cultured with hCG (0.1, 1, 10, 100 IU/ml), OP (10, 25, 50, 100 μM), RU486 (100 μM), P4 (1, 10, 25, 50 μM) or some combination of the four for 24 h. Cell viability was significantly increased by hCG (100 IU/ml) and significantly decreased by OP (100 μM) compared with the control. Cells treated with hCG and OP together were significantly less viable than the control and OP-treated cells. The combined treatment also significantly increased CASP3 activity and cleaved CASP3 protein expression. Furthermore, P4 addition reversed the reduction in cell viability caused by the combination of hCG and OP treatment. The overall findings suggest that hCG cooperates with P4 to increase survival of hLGCs and to induce apoptosis when P4 action supported by hCG is attenuated in the human CL.     

Detection of novel kobu-like viruses in Japanese black cattle in Japan

During surveillance for bovine diarrhea of unknown causes in Japanese black cattle in Kagoshima Prefecture, Japan, we found two types of novel kobu-like viruses in fecal samples of calves. Sequence analyses revealed that they had L protein and 2A protein with H-box/NC sequence motif, which are present in kobuviruses. Phylogenetic analysis revealed that they were related to kobuviruses; however, they clustered apart from other kobuviruses. In the prevalence study of two types of novel kobu-like viruses, 16.9% and 10.4% prevalence of these viruses were observed in the feces of diarrheal calves in this area.     

The luteotrophic function of galectin-1 by binding to the glycans on vascular endothelial growth factor receptor-2 in bovine luteal cells

The corpus luteum (CL) is a temporary endocrine gland producing a large amount of progesterone, which is essential for the establishment and maintenance of pregnancy. Galectin-1 is a β-galactose-binding protein that can modify functions of membrane glycoproteins and is expressed in the CL of mice and women. However, the physiological role of galectin-1 in the CL is unclear. In the present study, we investigated the expression and localization of galectin-1 in the bovine CL and the effect of galectin-1 on cultured luteal steroidogenic cells (LSCs) with special reference to its binding to the glycans on vascular endothelial growth factor receptor-2 (VEGFR-2). Galectin-1 protein was highly expressed at the mid and late luteal stages in the membrane fraction of bovine CL tissue and was localized to the surface of LSCs in a carbohydrate-dependent manner. Galectin-1 increased the viability in cultured LSCs. However, the viability of LSCs was decreased by addition of β-lactose, a competitive carbohydrate inhibitor of galectin-1 binding activity. VEGFR-2 protein, like galectin-1, is also highly expressed in the mid CL, and it was modified by multi-antennary glycans, which can be recognized by galectin-1. An overlay assay using biotinylated galectin-1 revealed that galectin-1 directly binds to asparagine-linked glycans (N-glycans) on VEGFR-2. Enhancement of LSC viability by galectin-1 was suppressed by a selective inhibitor of VEGFR-2. The overall findings suggest that galectin-1 plays a role as a survival factor in the bovine CL, possibly by binding to N-glycans on VEGFR-2.     

Mediastinal basaloid carcinoma arising from thymic cysts in two dogs

Mediastinal masses in dogs were diagnosed as basaloid carcinoma associated with multiple thymic cysts (MTCs). The masses were composed of MTCs and proliferating intracystic neoplastic basaloid cells, which immunohistochemically diffusely expressed p63 and cytokeratin 19. A gradual transition from the basal cell layers lining the cysts walls to the neoplastic cells was seen, and it was indicated that the neoplastic cells had originated from the basal cell layers of the cysts. To the best of our knowledge, this is the first report of basaloid carcinoma occurring in the mediastinal cavity in dogs. Although these tumors were demonstrated to be rare origins, basaloid carcinoma should be included in the differential diagnoses for canine mediastinal tumors.     

Sperm whales associated with Gulf Stream features off the north-eastern USA shelf

Marine mammal surveys were conducted during sum Mer 1990 and 1991 in shelf-edge and off-shelf waters between Cape Hatteras and Georges Bank. Sperm whales were the most frequently sighted large whale in both years, constituting 27.9% and 12.7% of total sightings in 1990 and 1991, respectively. Sighting rates of sperm whales that were within a 9.3 km buffer (5 n.m.) of the Gulf Stream north wall were not significantly different (x²= 2.86, P > 0.05) from those within the Gulf Stream during 1990. In 1991, however, significantly more (x²= 51.0, P < 0.05) sightings were associated with shelf-edge areas where warm core rings were located.     

Allele‐specific PCR typing and sequencing of the mitochondrial D‐loop region in four layer breeds

This study aimed to investigate the ability of single nucleotide polymorphism (SNP) haplotypes in chicken mtDNA for presumption of the origins of chicken meat. We typed five SNPs of the D‐loop region in mtDNA by allele‐specific PCR (AS‐PCR) in 556 hens, that is 233 White Leghorn (WL), 50 Dekalb‐TX35 (D‐TX), 140 Barred Plymouth Rock (BPR) and 133 Rhode Island Red (RIR) kept in the National Institute of Livestock and Grassland Science (NILGS, Tsukuba, Japan). Five haplotypes were observed among those chickens by AS‐PCR. WL, D‐TX, BPR and RIR displayed three, two, one and four SNP haplotypes, respectively. By a combination of the haplotypes by AS‐PCR and the breeds, these chickens were classified into 10 groups. After the D‐loop was sequenced in two chickens from every group (20 individuals), 15 SNP sites (including one insertion) and eight sequence haplotypes were observed. In conclusion, haplotype variation was observed in and among the layer breeds of the NILGS. This study demonstrates that SNP haplotypes in mtDNA should be appropriate for the presumption of the origins of chicken meat.     

Free beta-hydroxy-gamma-aminobutyric acid in brain

By using the methods of paper chromatography and high-potential paper electrophoresis and by comparing the pattern of prepared extracts with that of synthesized beta-hydroxy-gamma-aminobutyric acid, we found that free beta-hydroxy-gamma-aminobutyric acid exists in the brains of mice, rabbits, cattle, and human beings.     

Regression effect of carp, Cyprinus carpio L., peripheral blood lymphocytes on CHV-induced carp papilloma

Abstract. The effect of peripheral blood lymphocytes (PBL) of carp, Cyprinus carpio L., was investigated on the regression of papilloma which had been induced in carp experimentally infected with Herpesvirus cyprini (CHV). The regression of the papilloma was delayed in carp injected with anti-carp PBL serum. In addition, the spontaneous cytotoxic activity of PBL derived from normal carp was suppressed by injection with anti-carp PBL serum. These results suggest that the spontaneous cytotoxic activity of carp PBL plays an important role in the regression of carp papilloma.