Semi-quantitative immunohistochemical studies on Thy-1 antigen expressed by thymic myoid cells

Thy-1 antigen expression in the rat thymic myoid cell line R615B2 (Thy-1(+] and R613Ad (Thy-1(-] was studied with a semi-quantitative immunohistochemical assay without disrupting the cell cultures. With this assay, the quantity of Thy-1 antigen on R615B2 cells was detected separately in the cytoplasm and on the cell surface by the use of appropriate fixatives such as chilled ethanol and methanol + 0.3% H2O2. Extracellular Thy-1 antigen was also found in the culture supernatant of R615B2 cells. More than half of extracellular Thy-1 antigen remained in the supernatant even after 100,000 X g centrifugation. No form of Thy-1 antigen was detected at significant levels in R613Ad cells.     

Comparative studies on soluble protein profiles and isozyme patterns of seven Trichinella isolates

Soluble protein profiles and isozyme patterns of eight enzymes were compared for extracts of muscle stage larvae of the seven Trichinella isolates, using isoelectric focusing in polyacrylamide gel. Soluble protein profiles and isozyme patterns of four enzymes: malic enzyme, glucose-phosphate isomerase, phosphoglucomutase, superoxide dismutase of them were clearly divided into four types. T. pseudospiralis from a racoon and the Polar strain from a polar bear formed type 1 and type 2. The Iwasaki strain from a Japanese black bear and the Yamagata strain from a racoon dog, both from Japan, were type 3. Type 4 consisted of three remaining strains, the Polish strain from a wild pig, the USA strain from a pig and the Thai strain from a human case, which have similar infectivities to pigs. The Thai strain varied a bit electrophoretically from other members of type 4. Zymograms of adenylate kinase and malate dehydrogenase were similar in types 2 and 3. The 6-phosphogluconate dehydrogenase zymogram of type 3, similar to that of type 4, was different from that of type 2. It is assumed from the data that type 3 (Japanese strain) was genetically intermediate to types 2 and 4. T. pseudospiralis and the Polar strain had a common main isozyme of 6-phosphogluconate dehydrogenase. The zymogram of lactate dehydrogenase was common except for T. pseudospiralis.     

Characterization of adenocarcinoma of the lung in a familial adenomatous polyposis patient

The incidence of several extracolonic tumors, such as duodenal carcinoma, is higher in familial adenomatous polyposis (FAP) patients than in the general population, but there is little information about lung carcinoma in FAP. A 43-year-old woman presented with a lung tumor 17 years after total colectomy for FAP. Pathohistological analysis of the lung tumor demonstrated mixed adenocarcinoma consisting of a papillary adenocarcinoma component and a bronchioloalveolar carcinoma component. Sequencing analysis indicated a germline APC mutation from TCA to TGA (stop) at codon 1110, but no pathogenic germline MYH mutations. The other APC allele in the lung carcinoma was not inactivated by somatic mutations, promoter methylation, or chromosomal deletion. No somatic mutations in any of the coding regions of the p53 gene or in the mutation hot spot regions of the K-ras or EGFR genes were detected in the carcinoma. Amplification, however, of three chromosome regions, 5p, 8q, and 12q14-12q21, was identified in the carcinoma on genome-wide high-resolution single-nucleotide polymorphism (SNP) microarray. The present results suggest that the chromosomal copy number alterations detected on SNP microarray were involved in the carcinogenesis of the adenocarcinoma of the lung in the present FAP patient.     

A cryptic microdeletion including MBD5 occurring within the breakpoint of a reciprocal translocation between chromosomes 2 and 5 in a patient with developmental delay and obesity

The 2q23.1 deletion syndrome has been recently recognized as a neurodevelopmental disorder associated with intellectual disability, epilepsy, and autism spectrum disorder. Recently, methyl‐CpG‐binding domain 5 gene (MBD5), located in the 2q23.1 region, has been considered as a single causative gene of this syndrome. We report on a female patient with a de novo reciprocal translocation between chromosomes 2 and 5. Chromosomal microarray testing revealed a cryptic 896 kb deletion that included MBD5. Although clinical manifestations of this patient are compatible with those of patients with 2q23.1 deletion syndrome, a focal pachygyria revealed by brain magnetic resonance imaging has never been observed in the previously reported cases. Obesity caused by hyperphagia was observed in our patient and 28% of the previously reported patients with the 2q23.1 deletion syndrome. For better medical management, appropriate dietary guidance against hyperphagia should be given to the patients' family. © 2013 Wiley Periodicals, Inc.     

Partial response of angiosarcoma of the scalp to sorafenib: association with decreased expression of vascular endothelial growth factors and their receptors

The prognosis of angiosarcoma remains very poor, even with combined, multimodal therapy. We report a case with partial response of angiosarcoma of the scalp to sorafenib, which is a new oral, molecular, targeted, multiple-kinase inhibitor. In addition, we confirmed, using immunohistochemistry, that sorafenib suppressed the expression of vascular endothelial growth factors and their receptors on the angiosarcoma tumour cells, and decreased cell numbers by inhibiting cellular proliferation.     

Therapeutic Effects of Dipolar Aprotic Substances on Niemann-Pick Cells

Dimethylsulfoxide (Me₂SO) increased acid sphingomyelinase activity in cultured skin fibroblasts (Sakuragawa N et al., Biochem Biophy Res Comm 1985; 126: 756–762). Various drugs were examined to find related compounds showing the same phenomenon as Me₂ SO, including other dipolar aprotic substances, various inhibitors of DNA and RNA synthesis, phorbol esters, vitamin E and its derivatives and other miscellaneous agents. Several dipolar aprotic substances, such as Me₂SO, dimethylacetamide, tetramethylurea and hexa-methylene-bisacetamide, increased the acid and Mg-independent neutral sphingomyelinase activity in human cultured skin fibroblasts. These dipolar aprotic substances also increased acid and Mg-independent neutral sphingomyelinase activities in Niemann-Pick cells, type C, to 300 to 700% of those of untreated controls, repairing their enzyme deficiency. Other lysosomal hydrolases increased to a slightly lesser extent than sphingomyelinase. Dipolar aprotic substances could be used as therapeutic drugs for Niemann-Pick disease, type C.     

A practical enzyme immunoassay for anti-acetylcholine receptor antibodies in Myasthenia Gravis

A practical enzyme immunoassay (EIA) has been developed for the measurement of anti-acetylcholine receptor (AChR) antibodies in sera from patients with myasthenia gravis (MG). This system is based on the double antibody technique, using denervated rat muscle AChR labeled with horseradish peroxidase-linked α-bungarotoxin (HRP-αBGT). This method has the following advantages compared to conventional radioimmunoassay (RIA): (1) HRP-αBGT is more stable than [¹²⁵I]αBGT and can be used for at least one year without any loss of the binding activity to AChR and enzymatic activity, (2) the procedure avoids the use of radioactive isotopes, and (3) the equipment for our EIA is more economical than that for RIA.