Terminal differentiation and senescence share several common properties, including irreversible cessation of growth and changes in gene expression profiles. To identify molecules that converge in both processes, an overlapping pathway screening was employed that identified old-35, which is human polynucleotide phosphorylase (hPNPase old-35 ), a 3,5-exoribonuclease. We previously demonstrated that hPNPase old-35 is a type I interferon-inducible gene that is also induced in senescent fibroblasts. In vitro RNA degradation assays confirmed its exoribonuclease properties, and overexpression of hPNPase There are two contrasting endpoints in the life of a replicating cell. One involves the normal physiological processes of differentiation or senescence. The other is the pathological process of neoplastic transformation characterized by uncontrolled proliferation and de-differentiation. Treatment of HO-1 metastatic human melanoma cells with fibroblast interferon (IFN-) 1 and the protein kinase C activator mezerein (MEZ) induces irreversible growth arrest and terminal differentiation characterized by changes in cell morphology, increase in melanin synthesis, modifications in gene expression, and alterations in surface antigen expression (1-5). Replicative or cellular senescence, a process leading to irreversible arrest of cell division, was first described in cultures of human fibroblasts that lost the ability to divide upon continuous subcultures (6). Replicative senescence can result from telomere shortening linked with a DNA end-replication problem, overexpression of certain oncogenes, or tumor suppressor genes, or it can be stress-induced premature senescence after exposure to a variety of oxidative stresses or DNA damaging agents (for a review, see Ref. 7).Terminal differentiation and cellular senescence share several common traits including irreversible growth arrest and changes in gene expression profiles. To understand the molecular and biochemical basis of the complex physiological changes associated with these phenomena, an overlapping pathway screen was used to identify genes displaying coordinated expression as a consequence of both processes (8). A temporally spaced terminally differentiated human melanoma subtracted cDNA library was screened with cDNAs derived from senescent progeroid fibroblast cells. This led to the identification of old-35, which is human polynucleotide phosphorylase (hPNPase old-35 ), a 3Ј,5Ј exoribonuclease involved in RNA degradation (8). hPNPase old-35 is a highly evolutionary conserved gene in plants, prokaryotes and eukaryotes having similar domain structure and functional properties in all species. In vitro assays confirmed that hPNPase old-35 is involved in RNA degradation. Analysis of the expression profile of hPNPase old-35 revealed that it is predominantly a type I interferoninducible gene, and its expression is also induced in senescent fibroblasts in comparison with young fibroblasts. These findings indicate that hPNPase old-35 might play an essential role in
Human melanoma cells growth-arrest irreversibly and terminally differentiate on treatment with a combination of fibroblast interferon and the protein kinase C activator mezerein. This experimental protocol also results in a loss of tumorigenic potential and profound changes in gene expression. Various cloning and cDNA microarray strategies are being used to determine the complete spectrum of gene expression changes underlying these alterations in human melanoma cells. An efficient approach, Rapid Subtraction Hybridization (RaSH), has been developed that is permitting the identification of genes of potential relevance to cancer growth control and terminal cell differentiation. RaSH cDNA libraries are prepared from double-stranded cDNAs that are enzymatically digested into small fragments, ligated to adapters, and PCR amplified followed by incubation of tester and driver PCR fragments. This subtraction hybridization scheme is technically simple and results in the identification of a high proportion of differentially expressed sequences, including known genes and those not described in current DNA databases. The RaSH approach represents an efficient methodology for identifying and cloning genes displaying differential expression that associate with and potentially regulate complex biological processes.cDNA cloning ͉ reverse Northern blotting ͉ melanoma differentiation associated genes ͉ Northern blotting
Cu wire is drastically replacing Au wire due to surge of Au price. However, Cu wire package has poorer humidity reliability than Au wire package. Although Pd coated Cu wire package could show better humidity reliability than Cu wire, it is still worse than Au. Enough information regarding failure mechanism was not available. For failure analysis, x-section has been widely used to identify the Cu/Al IMC after failure. However, the x-section is the results of corrosion reaction and doesn't show the IMC status before corrosion. Therefore, the failure mechanism could not be estimated precisely. We used chemical model simulation to predict what kinds of IMC could be created after wire bonding, then which IMC could be corroded more easily during HAST. The Desorption energy was used to estimate reactivity between specified Cu/Al IMC and chlorine ion. The simulation suggested that the formation of Cu rich and Cu poor Cu/Al IMC and the Cu rich IMC was estimated to be corroded by chlorine ion. These chemical model simulations are the effective way to have fundamental understanding of the mechanism of Cu/Al IMC corrosion. Furthermore, chemical model simulation for Pd coated Cu wire was done to explore the effect of Pd existence and distribution of Pd in Cu/Al IMC. Dispersed Pd contributed to create new IMC of Cu/Al/Pd instead of easily corroded Cu rich Cu/Al IMC. Cu and Al diffusion and also Cl ion diffusion were inhibited by Pd at surface. Even Cl ion catching effect by Pd is also discussed.To improve humidity reliability performance with Cu wire, we developed new ion trapper using chemical model simulation technique. Developed molding compounds with the ion trapper showed significant improvement at bias HAST with Cu wire, which was even better than conventional Cu wire compatible molding compounds. IntroductionOne of the concerns to use Cu wire has been failure during humidity reliability test such as PCT, unbias HAST and bias HAST. [1,2] Failure mode of humidity reliability test was open with observed corroded layer and crack. We also confirmed that open failure at positive pad at bias HAST.[3] Conventional Br contained molding compound showed worse HAST results. However, even green molding compound showed HAST failure. We explored the factors of humidity reliability failure for Cu wire packages. As a result, extracted chlorine ion from molding compounds turned to be major factor while pH of extracted water turned to be minor factor through bias HAST. We also examined impact of wire bonding strength, bias and reliability condition.
The centipede Scolopendra subpinipes mutilans is a medicinally important arthropod species. However, its transcriptome is not currently available and transcriptome analysis would be useful in providing insight into a molecular level approach. Hence, we performed de novo RNA sequencing of S. subpinipes mutilans using next-generation sequencing. We generated a novel peptide (scolopendrasin II) based on a SVM algorithm, and biochemically evaluated the in vitro antimicrobial activity of scolopendrasin II against various microbes. Scolopendrasin II showed antibacterial activities against gram-positive and -negative bacterial strains, including the yeast Candida albicans and antibiotic-resistant gram-negative bacteria, as determined by a radial diffusion assay and colony count assay without hemolytic activity. In addition, we confirmed that scolopendrasin II bound to the surface of bacteria through a specific interaction with lipoteichoic acid and a lipopolysaccharide, which was one of the bacterial cell-wall components. In conclusion, our results suggest that scolopendrasin II may be useful for developing peptide antibiotics.
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