Molecular cloning, gene expression analysis, and recombinant protein expression of novel silk proteins from larvae of a retreat-maker caddisfly, Stenopsyche marmorata

Retreat-maker larvae of Stenopsyche marmorata, one of the major caddisfly species in Japan, produce silk threads and adhesives to build food capture nets and protective nests in water. Research on these underwater adhesive silk proteins potentially leads to the development of new functional biofiber materials. Recently, we identified four major S. marmorata silk proteins (Smsps), Smsp-1, Smsp-2, Smsp-3, and Smsp-4 from silk glands of S. marmorata larvae. In this study, we cloned full-length cDNAs of Smsp-2, Smsp-3, and Smsp-4 from the cDNA library of the S. marmorata silk glands to reveal the primary sequences of Smsps. Homology search results of the deduced amino acid sequences indicate that Smsp-2 and Smsp-4 are novel proteins. The Smsp-2 sequence [167 amino acids (aa)] has an array of GYD-rich repeat motifs and two (SX)(4)E motifs. The Smsp-4 sequence (132 aa) contains a number of GW-rich repeat motifs and three (SX)(4)E motifs. The Smsp-3 sequence (248 aa) exhibits high homology with fibroin light chain of other caddisflies. Gene expression analysis of Smsps by real-time PCR suggested that the gene expression of Smsp-1 and Smsp-3 was relatively stable throughout the year, whereas that of Smsp-2 and Smsp-4 varied seasonally. Furthermore, Smsps recombinant protein expression was successfully performed in Escherichia coli. The study provides new molecular insights into caddisfly aquatic silk and its potential for future applications. (C) 2015 Elsevier Inc. All rights reserved.ArticleBIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS. 464(3):814-819 (2015)journal articl

Anti-asialoglycoprotein receptor autoantibodies, detected by a capture-immunoassay, are associated with autoimmune liver diseases.

In autoimmune chronic active hepatitis (AIH) and primary biliary cirrhosis (PBC), various autoantibodies including anti-asialoglycoprotein receptor (ASGPR) antibodies have been found in patients' sera. We have previously developed a mouse monoclonal antibody against rat and human ASGPR. In this study, we developed a capture enzyme-linked immunosorbent assay (ELISA) for detection of anti-ASGPR antibodies using this monoclonal antibody and investigated the occurrence of anti-ASGPR antibodies in the sera of patients with various liver diseases. Serum samples were obtained from 123 patients with various liver diseases, including 21 patients with AIH and 40 patients with PBC. In this capture ELISA, the target antigen in the crude rat liver membrane extracts was captured on the ELISA wells by the ASGPR-specific mouse monoclonal antibody. Thus, the cumbersome process of antigen purification was rendered unnecessary. Using this capture ELISA, we detected the anti-ASGPR antibody in 67% of the patients with AIH, in 100% of the patients with PBC, and in 57% of the patients with acute hepatitis type A. However, the anti-ASGPR antibody was rarely detected in patients with other liver diseases such as primary sclerosing cholangitis and obstructive jaundice. Our findings suggest that this capture ELISA would be useful for the detection of anti-ASGPR antibodies in autoimmune liver diseases.</p

Ultrasound guided radio-frequency albation in human hepatocellular carcinoma

Hapatocellular carcinoma (HCC) is one of the most prevalent malignancies in Japan. Percutaneous ethanol injection therapy (PEIT) of microwave coagulation therapy (MCT) have been used to treat small HCC. These two methods are effective methods to treat HCC; however, they have weak points. A period of admission is long to treat patients with PEIT because it requires several sessions to kill HCC completely. The risk of complications of MCT is high because the needle is thick, although it can coagulate HCC completely in one session. Radio frequecy ablation (RFA) is a new method to treat HCC. RFA compensates these weak points and is thought to become a main modality to treat HCC. In this paper, we demonstrated two patients with HCC who were treated with RFA. The effect of the treatment was examined with helical computed tomography and contrast harmonic power doppler, and the results were satisfactory. We also mentioned the merits and demerits of RAF in Discussion

Ultrasound guided radio-frequency albation in human hepatocellular carcinoma

Hapatocellular carcinoma (HCC) is one of the most prevalent malignancies in Japan. Percutaneous ethanol injection therapy (PEIT) of microwave coagulation therapy (MCT) have been used to treat small HCC. These two methods are effective methods to treat HCC; however, they have weak points. A period of admission is long to treat patients with PEIT because it requires several sessions to kill HCC completely. The risk of complications of MCT is high because the needle is thick, although it can coagulate HCC completely in one session. Radio frequecy ablation (RFA) is a new method to treat HCC. RFA compensates these weak points and is thought to become a main modality to treat HCC. In this paper, we demonstrated two patients with HCC who were treated with RFA. The effect of the treatment was examined with helical computed tomography and contrast harmonic power doppler, and the results were satisfactory. We also mentioned the merits and demerits of RAF in Discussion