Nationwide surveillance of West Nile virus targeting mosquitoes and dead birds from April 2004 through March 2007 in Japan.

We conducted nationwide West Nile virus (WNV) surveillance targeting mosquitoes and dead birds to reveal whether the virus and its potential vectors are present in Japan. A total of 12 766 mosquitoes and 230 dead birds were collected in April 2004-March 2005 (the 2004-2005 period), 10 755 mosquitoes and 267 dead birds in April 2005-March 2006 (the 2005-2006 period), and 8624 mosquitoes and 245 dead birds in April 2006-March 2007 (the 2006-2007 period). The species of most of the mosquitoes collected over the 3 years were Culex tritaeniorhynchus (47.82%) and Anopheles sinensis (28.49%), and other species included Aedes albopictus (6.75%), the Culex pipiens group (Cx. pipiens pallens and Cx. pipiens molestus: 5.37%), Aedes vexans nipponii (2.54%), Armigeres subalbatus (1.08%), and Aedes japonicus (0.95%). As for the dead birds, most were Passeriformes (456 specimens), which included several crow species, and the other orders included Anseriformes, Columbiformes and Ciconiiformes (78, 66 and 36 specimens, respectively). All the specimens tested negative for WNV RNA by reverse-transcriptase polymerase chain reaction (RT-PCR) in the 2004-2005 period and by real-time RT-PCR in the 2005-2006 and the 2006-2007 periods, respectively. Our surveillance provided no evidence for WNV in Japan as of the end of the surveillance period, but on the other hand, it revealed that several species of potential WNV vectors are distributed widely in Japan, which suggests that WNV in principle could be transmitted by the potential vectors if introduced. Thus, it is essential to take continued precautions against WNV introduction.

Distribution of marine birnavirus in cultured marine fish species from Kagawa Prefecture, Japan.

To determine the distribution of marine birnavirus (MABV) in cultured populations of different marine fish species, 1291 pooled tissue samples from 2672 fish belonging to 22 species and one hybrid were collected from Kagawa Prefecture, Japan, during 1999-2001. Using cell-culture MABV was isolated from three species: yellowtail, Seriola quinqueradiata Temminck & Schlegel (positive number/sample number, 10/419), amberjack, S. dumerili (Risso) (4/72), and Japanese flounder, Paralichthys olivaceus (Temminck & Schlegel) (41/481). Using PCR on MABV-negative samples, the MABV genome was detected in the same three species [yellowtail (9/409), amberjack (4/68) and Japanese flounder (93/440)] and two additional species, spotted halibut, Verasper variegatus (Temminck & Schlegel) (5/11), and goldstriped amberjack, S. lalandi Valenciennes (1/5). These MABV-positive species can be taxonomically divided into two groups: the genus Seriola and flatfish. In Japanese flounder, MABV was detected during all seasons, and the infection rate was correlated with water temperature. Aquaculture sites with MABV-positive fish were evenly distributed over the surveyed area, suggesting that MABV is widely distributed at aquaculture sites in Kagawa Prefecture. The nucleotide sequence at the variable region, the VP2/NS junction, revealed that the 39th base mutation occurs host-specifically for flatfish. Flatfish are suspected to be the main reservoir of MABV and might be responsible for establishing the infection cycle in aquaculture environments.

Early phase of intestinal mantle cell lymphoma: a report of two cases associated with advanced colonic adenocarcinoma.

Intestinal mantle cell lymphoma characteristically produces multiple polyps, a finding reported as multiple lymphomatous polyposis. The early stages of intestinal mantle cell lymphoma before polyp formation and the pattern of initial lymph node invasion, however, have not been described. We recently encountered two cases of intestinal mantle cell lymphoma in their early development found incidentally associated with advanced colonic adenocarcinoma. We present herein the clinical, histopathological, immunohistochemical, and molecular genetic features of these two cases. In one case, a single polypoid mass was found with invasion limited to mucosa and submucosa of the terminal ileum and without lymph node compromise. In the second case, there were multiple mucosal aggregates of neoplastic cells without formation of polyps. Regional lymph nodes in the latter case showed either partial or complete involvement by lymphoma. In both cases, immunohistochemistry (CD20+, CD5+, cyclin D1+, CD10-, and CD23-), and demonstration of clonal immunoglobulin heavy chain and bcl-1 gene rearrangements by PCR analysis confirmed the diagnosis of mantle cell lymphoma.

Results of breast-conserving therapy for ductal carcinoma in situ: the Kyoto University experiences.

BACKGROUND: The purpose of this study was to evaluate the results of breast-conserving therapy (BCT), defined as the combination of breast-conserving surgery with axillary dissection and definitive radiation therapy for ductal carcinoma in situ (DCIS). METHODS: Between November 1987 and March 1998, 33 patients with DCIS undergoing BCT at our hospital were examined. The mean age was 48. All patients underwent quadrantectomy or wide excision as well as axillary dissection. Radiation therapy consisted of 50 Gy to the ipsilateral whole breast. Boost irradiation of 10 Gy was given to 15 patients with close or positive margins. Nearly all patients received adjuvant chemotherapy with 5-fluorouracil or its derivatives and adjuvant endocrine therapy with tamoxifen for 2 years. RESULTS: The minimum and median follow-up periods were 32 and 80 months, respectively. All patients but one were followed. Only one patient had a non-invasive local recurrence, 23 months after her operation. This patient was salvaged with simple mastectomy. Her prognostic index score was 8. The five-year local control rate was 97%. No serious acute or late complications were noted. CONCLUSION: The results of this retrospective study substantiate favorable data and appear to confirm the efficacy and reasonable local recurrence rate of BCT for the treatment of DCIS.

MAIL, a novel nuclear I kappa B protein that potentiates LPS-induced IL-6 production.

We have identified and characterized a novel member of the ankyrin-repeat family named 'molecule possessing ankyrin-repeats induced by lipopolysaccharide' (MAIL). The C-terminal portion of MAIL shared high sequence homology with the I kappa B family. Intraperitoneal injection of lipopolysaccharide (LPS) into mice rapidly (<0.5 h) induced MAIL mRNA in various tissues, particularly in the spleen, lymph node, and lung. Ectopically expressed MAIL was localized in the nucleus, and remarkably potentiated the LPS-induced mRNA expression and secretion of interleukin (IL)-6 in Swiss 3T3 cells. These findings indicate that MAIL is one of the nuclear I kappa B proteins and an activator of IL-6 production.

Sympathetic activation of hepatic and splenic IL-1beta mRNA expression during oscillation stress in the rat.

Interleukin (IL)-1beta mRNA expression in the liver and spleen was examined after subjection to oscillation stress in the rat. Thirty-minute subjection to oscillation stress increased IL-1beta mRNA expression in the both organs. Prior treatment of rats with gadolinium chloride, which eliminates macrophages, prevented the stress-induced IL-1beta expression. Either adrenalectomy or treatment of guanethidine, a blocker of norepinephrine release in the sympathetic nerve endings, partially attenuated the stress-induced response, but the combined treatment completely blocked it. Injection of beta-adrenergic antagonist (propranolol) also suppressed the stress-induced response. These results suggest that oscillation stress induces IL-1beta mRNA expression in the liver and spleen, probably in Kupffer cells and splenic macrophages, and that stress-induced IL-1beta expression is elicited by catecholamines released from sympathetic nerve terminals and the adrenal gland.

Induction of uncoupling protein (UCP) 2 in primary cultured hepatocytes.

Uncoupling protein 2 (UCP2) mRNA expression and function was examined in rat primary cultured hepatocytes. UCP2 mRNA was not expressed in freshly isolated hepatocytes, but appeared during a 24-144 h primary culture period. Isolated mitochondria from 144 h cultured hepatocytes showed a lower oxygen consumption rate in the presence of succinate and ADP. However, the ratio of the oxygen consumption rate when media contained succinate alone to that with succinate and ADP was increased by 166% versus control mitochondria. Moreover, the mitochondrial potential in the presence of succinate was decreased by 60%, indicating the potential role of UCP2 in hepatocyte mitochondria as an active uncoupler.

Living related liver transplantation: histopathologic analysis of graft dysfunction in 304 patients.

Between June 1990 and August 1997, 304 mainly pediatric patients underwent a total of 311 orthotopic living related liver transplantations (LRLTs) under tacrolimus immunosuppression at Kyoto University Hospital. Congenital biliary atresia was the most common underlying disease. The donor was a parent, and the left lateral segments were used as grafts in most cases. The average number of loci of HLA-A, -B, and -DR mismatches between the donor and the recipient were 2.1. Forty-three transplants were ABO-incompatible. Liver histology at the time of abnormal liver function after transplantation was analyzed. Preservation injury was rare and mild. Acute cellular rejection (ACR) occurred in 36% of transplants during the first 6 months. Average rejection activity index (the Banff schema) was 4.2 and severe rejection was rarely seen. The number of mismatching HLA loci and immunosuppression regimens affected the incidence of ACR. Chronic rejection (CR) occurred in 2% of transplants. Concerning humoral rejection, no hyperacute rejection was seen. However, hepatic artery thrombosis (delayed hyperacute rejection) was seen in an ABO-incompatible transplant. Acute hepatitis, including those related to cytomegalovirus and Epstein-Barr virus, occurred in 17% of transplants. Chronic hepatitis, including hepatitis B and C, developed in 3%. Acute or chronic cholangitis occurred in 16%, and a significantly higher incidence of cholangitis was found in ABO-incompatible transplants. Posttransplantation lymphoproliferative disease developed in 2%. In LRLT, milder preservation injury and less frequent ACR and CR were suggested, probably because of the short cold-ischemia time and the advantages of HLA histocompatibility, respectively.

Ovarian dysgerminoma showing high serum levels and positive immunostaining of placental alkaline phosphatase and neuron-specific enolase associated with elevation of serum prolactin level.

We report a 35-year-old Japanese female patient with ovarian dysgerminoma showing elevated serum levels of placental alkaline phosphatase (PLAP), neuron-specific enolase (NSE) and prolactin (PRL). All elevated tumor markers improved dramatically after the removal of the tumor. Immunohistochemically examined, the tumor was stained positive for PLAP and NSE and negative for PRL. Our present case is the first report of dysgerminoma showing positive immunostaining for PLAP and NSE, and the association of high serum level of PRL followed by decrease accompanied by the tumor debulking.

The origin of SL family mice.

The origin of SL family mice was studied by analyzing 100 microsatellite loci, the major histocompatibility complex, the Mx gene, murine leukemia provirus, and mammary tumor provirus. From the genetic profile of family members and their history, we assumed the existence of a proto-SL mouse, an ancestor of all SL family members. Many alleles were contributed to the proto-SL by the ancestors related to strains A2G and CF#1, and/or some wild mice. Among four existing family members, SL/Am and SL/Ni mice were almost identical and presumably closest to the proto-SL. The SL/Kh mouse was derived from a cross of the proto-SL and AKR mice, because SL/Kh mice inherited a considerable number of genes from AKR mice, the most outstanding of which were those of the provirus Emv-11 and Thy-1.1. The SL/QDj mice seemed to be a recombinant inbred strain between SL/Am and SL/Kh mice, because their alleles at all 100 microsatellite loci were shared by SL/Am or SL/Kh strains or both. All four SL family members shared the major histocompatibility complex haplotype q.

Highly sensitive and simple liquid chromatographic determination in plasma of B6 vitamers, especially pyridoxal 5'-phosphate.

A simple and sensitive high-performance liquid chromatographic method for measuring the major vitamers of vitamin B6, i.e. pyridoxal and pyridoxal 5'-phosphate (PLP), and 4-pyridoxic acid (4-PA) in plasma was developed. The vitamers and 4-PA from plasma were extracted with 0.8 mol/l perchloric acid. The separation by HPLC is accomplished using an ODS reversed-phase column and a mobile phase of 0.1 mol/l potassium dihydrogen phosphate containing 0.1 mol/l sodium perchlorate, 0.5 g/l sodium bisulfite adjusted to pH 3, at a flow-rate of 1.0 ml/min. The vitamers and 4-PA were eluted within 13 min and their concentration is determined with a fluorometric detector (excitation, 300 nm; emission, 400 nm). In this method, PLP in plasma can be determined with high sensitivity using derivatization with sodium bisulfite in the mobile phase.

The SMXA: a new set of recombinant inbred strain of mice consisting of 26 substrains and their genetic profile.

A new set of recombinant inbred (RI) strain SMXA consisting of 26 substrains was established between SM/J and A/J. The history of the SMXA RI strains and their genetic profile covering 158 genetic marker loci are reported. From the strain distribution pattern among SMXA RI strains, the chromosomal location of salivary and tear protein genes Spe1-r, Spe1-s, Spe2, and Tpe1 were newly determined.

K-ras and p53 alterations in genomic DNA and transcripts of human pancreatic adenocarcinoma cell lines.

We analyzed 15 human pancreatic adenocarcinoma cell lines for alterations of the K-ras and the p53 genes and their transcripts. In 11 cell lines (73.3%), point mutations of the K-ras gene were found at codon 12 in exon 1. In 9 cell lines one allele was mutated and the other was wild type, and both the alleles were expressed into mRNA. In one cell line both alleles of codon 12 were mutated to TGT and GTT, respectively, but only TGT was transcribed into mRNA. Alterations in mRNA of the p53 gene were detected in 10 cell lines (66.7%). Analysis of the genomic sequence of the p53 gene revealed that the alterations consisted of 6 cases of base pair substitutions and 1 case of 1-bp deletion in evolutionarily conserved exons 5 to 8, 2 cases of splicing mutations in exon 4, and 1 case of novel deletion from exons 2 to 9. In 14 cell lines (93.3%), alterations were identified in the K-ras or p53 gene. Of these, 4 cell lines harbored K-ras mutations without p53 alteration, whereas 3 cell lines exhibited p53 alterations without K-ras mutation. Thus, it is suggested that activation of the K-ras gene and inactivation of the p53 gene are strongly and cooperatively associated with pancreatic carcinogenesis.