Heterogeneity of Somatostatin-Like Immunoreactivities Extracted from Rat Small Intestine

The small intestines of 5 rats were successfully divided into the epithelial cells and the muscle layer, and substances with somatostatinlike immunoreactivity (SLI) were extracted to investigate the heterogeneity of rat intestinal SLI. Separation of SLI by gel chromatography revealed that there were large differences in the relative amounts of 3.5K SLI and 1.6K SLI between the epithelial cells and the muscle layer. The ratio of 3.5K SLI / (3.5K SLI + 1.6K SLI) was 67.3±4.0% (Mean±SD, 54.2 - 81.0%) in the epithelial cells and 14.2±2.6% (Mean±SD, 2.8 - 18.8%) in the muscle layer. By High Performance Liquid Chromatography (HPLC), 3.5K SLI and 1.6K SLI were shown to correspond to somatostatin-28 (SS-28) and somatostatin-14 (SS-14), respectively. Thus it was demonstrated that SS-28 is predominantly distributed in the epithelial cells, and the SS-14 is mainly located in the muscle layer of rat small intestine

Carrier PNA for shRNA delivery into cells

A peptide nucleic acid (PNA)-cell-penetrating peptide (CPP) conjugate (carrier PNA) was used as 'bridgebuilder' to connect a CPP with an shRNA. The carrier PNA successfully formed a hybrid with an shRNA bearing complementary dangling bases and the shRNA was introduced into cells by the carrier PNA, and RNAi was induced by the shRNA

Antisense effect of pyrrolidine-based oxy-peptide nucleic acids in Escherichia coli

To investigate the antisense effect of a pyrrolidine-based oxy-peptide nucleic acid (POPNA), we carried out the LacZ reporter assay using a 12-mer trans-l-POPNA conjugated with a cell-penetrating peptide (antisense reagent). The antisense effect of the conjugated POPNA (inhibition of LacZ activity) was comparable to that shown by a Nielsen-type peptide nucleic acid. Furthermore, the conjugated POPNA could switch the LacZ activity over a wide range of ambient temperatures

A Radioimmunoassay for Rat Serum Corticosterone

A reliable radioimmunoassay for rat serum corticosterone has been developed. 25 μ1 of diluted serum (1:100) was assayed with a specific antiserum raised against corticosterone-21-hemisuccinate conjugated to bovine serum albumin. The within-assay and between-assay coefficients of variation were 7.1% and 13.9%, respectively. The mean serum corticosterone concentration was 65.3±6.0 ng/ml (n=10). The corticosterone level increased to 208.4 ±23.7 ng/ml after ACTH administration, and was suppressed to the limit of assay sensitivity after dexamethasone administration

Steroidogenesis in Isolated Adrenal Cells of Rat

A reliable and reproducible system for the isolation of rat adrenal cells was developed, using 0.25% trypsin for cell dispersion. The suspending cell in Krebs-Ringer bicarbonate buffer containing 0.2% glucose and 0.5% bovine serum albumin was incubated for 120 minutes at 37℃ under 95% 02 and 5% C02. Corticosterone production induced by synthetic 1-24ACTH showed a dose-related increase in decapsular cells. The precision of the inter-experiment of corticosterone production was 5.0% (average coefficients of variation)

Analysis of Human Insulin Analogues in Vitro, Using Gel Chromatographic Method

The incubation medium and incubated human pancreas were gel chromatographed on the Bio-Gel P-30 column after extraction with acid ethanol. The extracted immunoreactive insulin (IRI) was successfully separated into two peaks at the position of 6000 molecular weight region. These two peaks corresponded to those which were detected in human serum. These findings suggest that the two groups of insulin are directly secreted from human pancreatic tissue. But the incorporated [3H] leucine peak into acid-ethanol extractable protein did not elute out at the same position as each insulin peak. Therefore, the measurement of [3H] leucine incorporation into acid-ethanol extractable protein is not a good indicator to evaluate insulin biosynthesis

New bleeding model of additives in a polypropylene film under atmospheric pressure

Many additives are commercially used to add more favorable qualities to films. The bleeding process by which the additive in a film comes to the surface is considered. A new bleeding model of additives in a polypropylene film under atmospheric pressure was investigated. Solubility and diffusion are found to be important for explaining this bleeding process. The solubilities and diffusion coefficients of higher fatty acid amides such as erucamide (13-cis-docosenamide) and behenamide (docosanamide) were determined between 40 and 70°C and the difference between the solubilities and the diffusion coefficients was discussed. The experimental results are explained more precisely by assuming two transport processes between the crystalline regions and the amorphous ones. © 2007 Wiley Periodicals, Inc

New bleeding model of additives in a polypropylene film under atmospheric pressure II

金沢大学大学院自然科学研究科出光興産㈱　先進技術研究所　解析技術センター 第二解析技術室Many additives are commercially used to add more favorable qualities to films. The bleeding process by which the additive in a film comes to the surface is considered. A new bleeding model of additives in a polypropylene film under atmospheric pressure was investigated. Solubility and diffusion are found to be important for explaining this bleeding process. It was found that the experimental results were explained more precisely by assuming a twostep transport process between the crystalline regions and the amorphous ones. The solubilities and diffusion coefficients of UV-stabilizers such as 2-(2H-benzotriazol-2-yl)-4(l,l,3,3-tetramethylbutyl)phenol and 2-(2H-benzotriazol-2yl)-4-methylphenol were determined at 4O°C. The difference between the saturation solubilities and the diffusion coefficients of UV-stabilizers was discussed by comparing with the results of molecular dynamics (MD) simulation. © 2007 Wiley Periodicals, Inc