Sendai virus (SeV), a pneumotropic virus of rodents, has an accessory protein, V, and the V protein has been shown to interact with MDA5, inhibiting IRF3 activation and interferon-β production. In the present study, interaction of the V protein with various IRF3-activating proteins including MDA5 was investigated in a co-immunoprecipitation assay. We also investigated interaction of mutant V proteins from SeVs of low pathogenicity with MDA5. The V protein interacted with at least retinoic acid inducible gene I, inhibitor of κB kinase epsilon and IRF3 other than MDA5. However, only MDA5 interacted with the V protein dependently on the C-terminal V unique (Vu) region, inhibiting IRF3 reporter activation. The Vu region has been shown to be important for viral pathogenicity. We thus focused on interaction of the V protein with MDA5. Point mutations in the Vu region destabilized the V protein or abolished the interaction with MDA5 when the V protein was stable. The V-R 320 G protein was highly stable and interacted with MDA5, but did not inhibit activation of IRF3 induced by MDA5. Viral pathogenicity of SeV is related to the inhibitory effect of the V protein on MDA5, but is not always related to the binding of V protein with MDA5. Key words MDA5, Sendai virus-V protein.SeV, which belongs to the genus Respirovirus in the family Paramyxoviridae, is a respiratory tract pathogen of rodents. It is an enveloped virus with a single-stranded, negative-sense RNA genome of approximately 15.4 kilobases. The SeV genome comprises six genes encoding structural proteins, including N (nucleocapsid), P (phospho-), M (matrix), F (fusion), HN (hemagglutininneuraminidase), and L (large) proteins (1).The P gene, unlike the other genes, encodes not a single protein but multiple proteins. The colinear transcript encodes the P protein as well as C' , C, Y 1 and Y 2 proteins; the latter four proteins are translated in a shifted frame by alternative translational starts and a common stop codon. The P gene also directs synthesis of an additional mRNA for the V protein by inserting a pseudotemplated G residue at the specific editing site (2, 3). Consequently, P and V proteins share the same 317 residues at the amino terminus (P/V common region), while the two proteins have unique carboxyl termini. The V protein contains a 67-residue unique carboxyl terminus (Vu region), which is 760

In the pathogenesis of intestinal ischemia-reperfusion injury, the measurement of lipid peroxides needs to be established. Sprague-Dawley rat intestines were assessed after 30 minutes of occlusion of the superior mesenteric artery followed by reperfusion at 30, 60, 120, 180, 360 minutes. Grade of the mucosal injury, accumulation of the activated neutrophils and ICAM-1 expression were transiently increased after reperfusion. Two measuring methods of mucosal lipid peroxides using thiobarbituric acid reacting substance (TBARS) and phosphatidylcholine hydroperoxide (PCOOH) were compared. PCOOH level was significantly increased after reperfusion, while the mucosal TBARS level showed no significant change. In conclusion, lipid peroxidation could be detected with high specificity and sensitivity by measuring the mucosal phosphatidylcholine hydroperoxide level.

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Background: By definition, ductal carcinoma in situ (DCIS) does not metastasize to the lymph nodes. However, since the introduction of molecular whole-node analysis using the one-step nucleic acid amplification (OSNA) assay for sentinel node (SN) biopsies, the number of DCIS patients with SN metastasis has increased. The clinical management of node-positive DCIS remains controversial because these patients can be treated as different stages based on the pathogenesis: e.g. occult invasive cancer with true nodal metastasis (T1N1) or true DCIS with iatrogenic dissemination of benign or tumor cells into lymph node (TisN0). In this retrospective cohort study, we aimed to elucidate the pathogenesis of nodal metastasis in DCIS and the clinical management of node-positive DCIS. Patients and Methods: Subjects comprised of 427 patients with a routine postoperative diagnosis of DCIS who underwent SN biopsy using the OSNA assay between 2009 and 2012. The cut-off values of the OSNA assay for negative/positive results and micro/macrometastasis were defined at 250 and 5,000 copies/μL of cytokeratin 19 mRNA, respectively. In the SN-positive patients, all paraffin blocks containing the primary tumor were step-sectioned with 0.5-mm intervals until the tissue was exhausted, and all microscopic slides were examined for detecting occult invasions. Afterwards, the patients were classified into three cohorts based on the SN status and occult invasion: (1) no SN metastasis (TisN0), (2) SN metastasis without occult invasion (TisN1), and (3) SN metastasis with occult invasion (T1N1). Tumor characteristics including risk factors of occult invasions (e.g. large size, comedo-type), prognosis, and SN and non-SN status were compared among the three cohorts. The median follow-up time was 73.6 months. Results: Of the 427 patients, 408 (95.6%) were SN-negative and 19 (4.4%) were SN-positive. By examining a total of 1,421 step-sectioned slides, 9 of the 19 SN-positive patients had occult invasions in the primary tumors. Overall, 408 (95.6%), 10 (2.3%), and 9 (2.1%) were classified into the TisN0, TisN1, and T1N1 cohorts, respectively. Either of adjuvant endocrine therapy or chemotherapy was given much more in the TisN1 and T1N1 cohorts than in the TisN0 cohort (80.0% and 88.9% vs. 5.4%).Other tumor characteristics were similar among the three cohorts. Although one patients had distant recurrence in the TisN0 cohort, none had locoregional or distant recurrences in the TisN1 and T1N1 cohorts. Regarding the lymph node status in the TisN1 and T1N1 cohorts, median tumor burdens in the SN are 590 and 310 copies/μL, and 2 (20.0%) and 2 (22.2%) patients had additional non-SN metastasis in the axillary dissection materials, respectively. Conclusions: Tumor characteristics and prognosis were similar among the three cohorts albeit the TisN1 and T1N1 cohorts tended to received adjuvant systemic therapy. Moreover, the SN and non-SN status were similar between the TisN1 and T1N1 cohorts. Therefore, pathogenesis of nodal metastasis in DCIS cannot uniformly be explained, and tumors with different stages may be mixed in the node-positive DCIS. Thus, considering the favorable prognosis of node-positive DCIS, the clinical management should be determined on a case-by-case basis. Citation Format: Yonekura R, Osako T, Iwase T, Ogiya A, Ueno T, Ohno S, Akiyama F. Prognostic impact and possible pathogenesis of lymph node metastasis in ductal carcinoma in situof the breast [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P5-18-11.