Fatigue strength of steel pipe-base plate connections

The comparative fatigue resistance of two types of steel light poles is investigated, both experimentally and analytically. The fatigue tests were performed on test specimens designed and proportioned by the California Department of Transportation. The constant amplitude fatigue behaviour wa was obtained at several levels of stress range. Exposed fatigue crack surfaces were studied theascertain the nature of the crack initiation and propagation. Theoretical fatigue life estimates were also made using an existing fracture mechanics model, thought to simulate the geometric condition represented by the wclded pipe-base plate connection. The fatigue resistance of the two series of specimens was much lower than originally anticipated. The fatigue resistance was found to be comparable to either category E or E', depending on the weld contact angle.     

Fusion protein of interleukin-6 and interleukin-6 receptor without a polypeptide linker

FP6, a novel recombinant fusion protein of interleukin-6 (IL-6) and IL-6 receptor (IL-6R), was prepared in the methylotrophic yeast Pichia pastoris. This protein was a potent activator of a cell surface transducing glycoprotein, gp130 and is a potential therapeutical reagent in the hemopoietic field. A linker is generally thought to be required for two fused molecules to retain their proper structures although it should preferably be removed to reduce possible antigenicity. It was found that the C-terminal residue of IL-6R could be directly linked to the N-terminal residue of IL-6 without decreasing the ability of IL-6 to bind gp130 and send the IL-6 signal. It was also found that the peptide bond between Lys-37 and Asp-38 of IL-6 was prone to proteolytic cleavage and that the immunoglobulin (Ig)-like region of IL-6R underwent extensive and heterogeneous glycosylation when expressed in P. pastoris. Based on these findings, we designed FP6 without the Ig-like region, in which the C-terminal residue of Ala-333 of IL-6R was directly linked to Asp-38 of IL-6 by a peptide bond. Purified FP6 had both an in vitro effect on hemopoietic progenitors to generate various colonies and an in vivo effect on megakaryocyte progenitors to increase platelet counts. Four purified FP6s were obtained, which had the same molecular mass and different isoelectric points without any detectable modification in the course of purification. The difference in isoelectric points was shown to be due to microheterogeneity of the carbohydrate chains. Each FP6 had the same specific activity in the cell growth assay with or without endoglycosidase digestion. Homogeneous FP6 with respect to isoelectric point as well as molecular mass merits more detailed characterization and evaluation for possible clinical application.     

Extractability of antioxidants from legume seed flour after cooking and in vitro gastrointestinal digestion in comparison with methanolic extraction of the unprocessed flour

Antioxidant activities were studied in methanolic and water extracts of nonprocessed, cooked and in vitro enzymatically digested seed flour, as well as in total protein hydrolysates and small peptide fractions (<3 and <10 kDa) of three pea and five grass pea cultivars. The antioxidative properties were determined by three spectrophotometric methods: 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical scavenging assay, Folin‐Ciocalteu (FC) reducing capacity assay and H₂O₂ scavenging. We also applied one luminometric assay for hydroxyl radical scavenging. The study showed that cooking and enzymatic digestion strongly enhanced the release of phenolic compounds in methanolic extracts of four analysed cultivars. Scavenging activity against DPPH radical, hydroxyl radical and hydrogen peroxide was increased in majority of analysed flour specimens subjected to processing. Our findings indicate that, besides the phenolic compounds, the small peptide fraction, especially the MW <3 kDa, in methanolic and aqueous extracts of cooked and digested seed flour significantly contribute to free radical and hydrogen peroxide scavenging activity in all investigated cultivars. Our data strongly suggest that simple cooking treatment and in vitro digestion of seed flour applied prior to extraction with methanol could improve antioxidative activity of obtained extracts.     

Effects of onion extract containing concentrated cysteine sulfoxides on sleep quality: a randomized,double-blind,placebo-controlled,crossover study

Food Science and Biotechnology - The objective of this randomized, double-blind, placebo-controlled, crossover study was to examine the effects of onion extract containing concentrated cysteine...     

MicroRNA (miRNA) cloning analysis reveals sex differences in miRNA expression profiles between adult mouse testis and ovary

MicroRNAs (miRNAs) are endogenous non-coding small RNAs that can regulate the expression of complementary mRNA targets. Identifying tissue-specific miRNAs is the first step toward understanding the biological functions of miRNAs, which include the regulation of tissue differentiation and the maintenance of tissue identity. In this study, we performed small RNA library sequencing in adult mouse testis and ovary to reveal their characteristic organ- and gender-specific profiles and to elucidate the characteristics of the miRNAs expressed in the reproductive system. We obtained 10,852 and 11 744 small RNA clones from mouse testis and ovary respectively (greater than 10,000 clones per organ), which included 6630 (159 genes) and 10,192 (154 genes) known miRNAs. A high level of efficiency of miRNA library sequencing was achieved: 61% (6630 miRNA clones/10,852 small RNA clones) and 87% (10,192/11,744) for adult mouse testis and ovary respectively. We obtained characteristic miRNA signatures in testis and ovary; 55 miRNAs were detected highly, exclusively, or predominantly in adult mouse testis and ovary, and discovered two novel miRNAs. Male-biased expression of miRNAs occurred on the X-chromosome. Our data provide important information on sex differences in miRNA expression that should facilitate studies of the reproductive organ-specific roles of miRNAs.     

Progesterone, but not 17beta-estradiol, up-regulates erythropoietin (EPO) production in human amniotic epithelial cells

Human amniotic epithelial (HAE) cells have great potential for successful use in cell therapy, since they do not cause acute rejection upon allotransplantation. However, to date, HAE cells have not well been studied. We previously reported that HAE cells produce erythropoietin (EPO), which is known to be a regulator of hematopoiesis, and that the induction mechanism of HAE cells is unknown, although EPO production from HAE cells is not increased by hypoxia which induces several cell types to produce EPO. In this study, we determined whether female sex hormones, including progesterone and 17beta-estradiol, affect the EPO production of HAE cells. Bioactive measurement of EPO activity in the culture supernatants of HAE-SV40 cells, which were immortalized by transfection with a simian virus 40 large T antigen, revealed that EPO bioactivity was significantly increased by treatment with progesterone, but not 17beta-estradiol. Treatment of HAE-SV40 cells with progesterone transiently increased the EPO mRNA level by fivefold, while there was no change in response to 17beta-estradiol. Furthermore, the progesterone receptor (PR)-B was detected in both HAE cells and HAE-SV40 cells by Western blotting. These results suggest that EPO synthesis in HAE-SV40 cells is stimulated by progesterone, but not by 17beta-estradiol, and thus it is highly likely that the EPO synthesis of HAE cells is also regulated by progesterone.     

Mutational analysis and localization of the inositol transporters of Saccharomyces cerevisiae

Saccharomyces cerevisiae possesses two inositol transport proteins, Itr1p and Itr2p, encoded by the ITR1 and ITR2 genes, respectively. Itr1p and Itr2p are high and low affinity transporters, respectively. Eight out of nine cysteine residues in Itr1p, which are common in two transporters, were converted to serine residues by site directed mutagenesis. All mutant genes suppressed the growth defect caused by itr1 disruption, indicating that cysteine residues are not essential for its function. Chimeric genes that express Itr1p and Itr2p fused to the green fluorescent protein (GFP) under the control of the ADH1 promoter were constructed. Both genes were functional. Fluorescence microscopy analysis indicated that both GFP-fused Itr1p and Itr2p are localized to the plasma membrane. A multi-copy plasmid that expresses GFP-fused Itr1p under the control of the original ITR1 promoter was constructed. Under inositol-free culture conditions, GFP-fused Itr1p appeared and was localized to the plasma membrane. When the cells were cultured in the presence of inositol, GFP-fused Itr1p gradually disappeared from the plasma membrane, the fluorescence being redistributed within the cell. Prolonged culture of the cells also caused the relocalization of transporter proteins. These results clearly indicate that the cellular relocalization of transport proteins is responsible for a reduction of inositol transport activity, which is caused by the presence of inositol in the medium or culturing of cells to the stationary phase.     

Degradation behavior of mechanically fastened joints of glass-reinforced crosslinked polyester immersed in hot water

This study deals with the degradation of mechanically-fastened GFRP joints immersed in hot water (80°C). The material used was randomly oriented E-glass fiber continuous strand mat with a crosslinked polyester. Three kinds of joint geometries were adopted; thickness was 3 mm, hole diameter was 6 mm, the distance from hole center to top-edge was 18 mm (3e), and specimen widths were 18 (3w), 30 (5w), and 42 mm (7w). Failure modes of original dry specimens were a function of joint geometry, The dominant failure mode of 3w3e joints was net-tension, whereas 5w3e and 7w3e joints displayed bearing failure. As degradation progressed, the dominant failure mode gradually shifted from net-tension to bearing failure. Strength reduction was marked in 5w3e and 7w3e joints, in which the dominant failure mode was bearing. Joint strength and failure mode were predicted from the combination of a macroscopic failure criterion and characteristic curves obtained from tensile testing of rectangular specimens with holes, bearing tests, and finite element analysis. Predictions agreed with experiment.