Molecular cloning and sequence analysis of the porcine precursor of endothelin-2

The amino acid sequences of two of the three endothelin (ET) family peptides, ET-1 and ET-3, are identical among mammals, whereas for the other family member, ET-2 or vasoactive intestinal contractor (VIC), the mouse and rat sequences differ from the human counterpart ET-2 by one amino acid residue. To examine more deeply the structural diversity among ET-2/VIC orthologs (EDN2), we screened porcine ET-2/VIC-like cDNAs using the 5' rapid amplification of cDNA ends (RACE) method with degenerate primers based on ET-2/VIC mature peptides. Sequence analysis of the cDNAs showed that ET-2 is present in pig. The full-length cDNA sequence, produced by combining 5' RACE and 3' RACE products, revealed the porcine precursor protein of ET-2 (PPET-2). Porcine PPET-2, made up of 214 amino acids, includes a 26-residue putative signal sequence, big ET-2, mature ET-2, and ET-2-like peptide. The percent sequence identity of porcine PPET-2 with human PPET-2, and rat or mouse precursor protein of VIC runs between approximately 70% and 74%. ET-2, although expressed in intestine, has no anti-microbial activity.     

Preparation and clinical application of indomethacin gel for medical treatment of stomatitis

The preparation and clinical applications of indomethacin (IM) gel were investigated in the treatment of stomatitis resulting from chemotherapy and radiotherapy for cancer. IM gel was prepared by adding various water-soluble polymers [hydroxypropyl cellulose (HPC), etc.] to IM aqueous solution. The release rate of IM from IM gels was found to decrease with increasing polymer concentration and viscosity and to follow a first-order reaction rate equation. The release rate of IM from the IM gel with HPC was decreased gradually with increasing polymer concentration and to be easily controllable compared with gels with other polymers. The time before pain relief occurred after application of the IM gel was slightly shorter and the duration of pain relief was longer compared with the IM aqueous solution. It was confirmed that IM gel is useful in the treatment of stomatitis.     

Japanese Society of Medical Oncology Clinical Guidelines: Molecular Testing for Colorectal Cancer Treatment,Third Edition

The Japanese Society of Medical Oncology (JSMO) previously published 2 editions of the clinical guidelines: “Japanese guidelines for testing of KRAS gene mutation in colorectal cancer” in 2008 and “Japanese Society of Medical Oncology Clinical Guidelines: RAS (KRAS/NRAS) mutation testing in colorectal cancer patients” in 2014. These guidelines have contributed to the proper use of KRAS and RAS mutation testing, respectively. Recently, clinical utility, particularly for colorectal cancer (CRC) patients with BRAF V600E mutation or DNA mismatch‐repair (MMR) deficiency, has been established. Therefore, the guideline members decided these genetic alterations should also be involved. The aim of this revision is to properly carry out testing for BRAF V600E mutation and MMR deficiency in addition to RAS mutation. The revised guidelines include the basic requirements for testing for these genetic alterations based on recent scientific evidence. Furthermore, because clinical utility of comprehensive genetic testing using next‐generation sequencing and somatic gene testing of analyzing circulating tumor DNA has increasingly evolved with recent advancements in testing technology, we noted the current situation and prospects for these testing technologies and their clinical implementation in the revised guidelines.     

A case of primary small intestinal cancer accompanied by virchow lymph node metastasis undergoing TS-1 treatment

A 72-year-old female was admitted to our hospital with the complaint of left neck lymph node swelling. Abdominal computed tomography (CT) revealed wall thickening of the small intestine and multiple lymph node metastases. Barium meal study of the small intestine showed circular stenosis. The patient was operated on under a diagnosis of tumor of the small intestine and left neck lymph node swelling. Needle biopsy of the left neck lymph node and partial resection of the small intestine was done without regional lymph node dissection because of Virchow lymph node metastasis. On the resected material a 5 x 4 cm type 2 tumor was identified. Pathological findings included poorly-differentiated adenocarcinoma, si (bladder), n 4, P 0, ly 3, v 3, H 0, M(-), Stage IV. The patient received the chemotherapy with TS-1. TS-1(80 mg/body/day) orally administered for 4 weeks followed by a drug-free 2-week period as one course. CT revealed that the metastatic lesion had shrunk markedly after the second course. A complete response (CR) was observed after one year. There were no drug side effects. At present, 3 years and 9 months after the operation, cervical and abdominal CT reveals no evidence of enlargement of the cervical and intraperitoneal lymph nodes.     

Graft-versus-ATLL effect induced by abrupt discontinuation of immunosuppression following allogeneic bone marrow transplantation

A 46-year-old man was admitted to our hospital with swelling of a neck lymph node in June, 2002, and was diagnosed with adult T-cell leukemia/lymphoma (ATLL). As ATLL cells were detected in the peripheral blood after two courses of multi-agent chemotherapy (LSG 15), the treatment was changed to biweekly CHOP therapy. After two courses, hematological remission was achieved. Allogeneic bone marrow transplant (allo-BMT) from HTLV- negative and HLA-matched sibling donor was performed (conditioned with cyclophosphamide 60 mg/kg x 2 and total body irradiation 12 Gy). Cyclosporine A (CsA) and short-term methotrexate (MTX) were used for graft-versus-host disease prevention. Though the HTLV- provirus DNA (Southern blot) disappeared, HTLV-I provirus DNA (real-time PCR) T-cell receptor ygammachain gene rearrangement DNA (Southern blot) were detected in bone marrow after allo-BMT. MRD disappeared after the withdrawal of CsA. After the allo-BMT transplant, a graft-versus-ATLL (GVATLL) effect may be induced by abrupt discontinuation of immunosuppression.     

Immunohistological diagnosis of plasma cell myeloma based on cytoplasmic kappa/lambda ratio of CD138-positive plasma cells

Abstract For differentiating reactive plasmacytosis from clonal plasma cell neoplasms such as plasma cell myeloma (PCM), it is important to determine the expression of the cytoplasmic light chain accurately. Through retrospective analysis, we studied the cytoplasmic kappa/lambda ratio of CD138-positive plasma cells in bone marrow from 50 patients with PCM and 50 controls by immunohistological analysis. The percentage of cytoplasmic light chain immunoreactive cells out of the total plasma cell population was shown by a novel quantitative image analysis approach using an Aperio ScanScope CS and the Membrane v9 algorithm. PCM cells were distinguished from normal plasma cells by cut-off levels between 0.35 and 5.5, a sensitivity of 100% and a specificity of 98.0%. Detection of the cytoplasmic kappa/lambda ratio of CD138-positive plasma cells could be a useful tool for simple, efficient and accurate diagnosis of PCM.     

Angiographic evaluation of hepatic arterial damage after transarterial chemoembolization for hepatocellular carcinoma

PURPOSE: The aim of this study was to assess the incidence, degree, and predictors of hepatic arterial damage (HAD) after transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: A total of 33 patients with unresectable HCC underwent TACE alone using a mixture of iodized oil, epirubicin, and gelatin sponge. A follow-up angiogram was available for 76 of 109 sessions, and HAD was evaluated at each subsegment of the hepatic artery using a three-grade scale (1, no or slight wall irregularity; 2, overt stenosis; 3, occlusion). Grades 2 and 3 were considered to indicate significant HAD. The predictors of HAD were analyzed by multivariate analysis. RESULTS: A total of 161 hepatic arteries were embolized from the lobar (n = 43), segmental (n = 40), subsegmental (n = 72), or more distal (n = 6) level. The follow-up period between the initial and last sessions ranged from 70 to 1505 days (median 497 days). Significant HAD occurred in 37 of 231 subsegmental hepatic arteries (16%) and in 16 of 33 patients (48%). The accumulated dose of epirubicin per artery (P = 0.001) and Child-Pugh score (P < 0.001) were significant predictors. CONCLUSION: TACE is more likely to induce HAD in cirrhotic patients with impaired liver function and when a high dose of the chemotherapeutic agent was used.