RESUMO
The surface hydrophilization of mixed plastic waste aggregates (MPAs) was conducted to improve the bond between an MPA and the surrounding cement matrix using two types of coating agents: a silicone amine resin and acrylic binders. The coating agents formed a physical bond with the MPAs, and the results of contact angle measurement also revealed that the surface of MPAs was hydrophilic. The workability of a mortar mix increased by up to 1.47 times with the surface hydrophilization of MPAs. Meanwhile, the compressive and flexural strengths of mortar mixes decreased by 29~43% and 72~86%, respectively, at 28 days with the surface hydrophilization of MPAs. Namely, the surface hydrophilization of MPAs was successively conducted, and the workability of mortar mixes was improved accordingly, but the compressive and flexural strengths of mortar mixes decreased as the physical bond was partially separated from not only the MPA but also the surrounding cement matrix and the surface friction was decreased.
RESUMO
Porcine pancreas development is not well studied at the molecular level despite being a therapeutic resource for diabetic patients. In this study, we investigated expression of lineage markers and performed proteomic analysis. Expression of the early lineage markers Pdx1 and Ptf1a was developmentally conserved between mice and pigs, whereas expression of the islet differentiation marker Pax4 was delayed in porcine compared with murine pancreas development. Proteomic analysis found that expression levels of chymotrypsinogen were downregulated during porcine pancreas development while those of digestive enzymes like lipases, elastase and serine protease were up-regulated. In addition, specific isoforms of protein folding assistants such as protein disulfide isomerase and prefoldin were expressed at specific stages during the maturation of digestive enzymes. Taken together, these results show that development of the porcine pancreas is regulated by a concerted interplay of pancreas lineage marker proteins and other specified proteins, resulting in a functional endocrine and exocrine organ.
Assuntos
Pâncreas/embriologia , Pâncreas/metabolismo , Proteômica , Sus scrofa/embriologia , Sus scrofa/metabolismo , Animais , Biomarcadores/metabolismo , Linhagem da Célula , Eletroforese em Gel Bidimensional , Inibidores Enzimáticos/metabolismo , Precursores Enzimáticos/metabolismo , Pâncreas/enzimologia , Dobramento de Proteína , Proteínas/classificação , Proteínas/metabolismoRESUMO
The ability to generate neural lineages from human embryonic stem cells (hESCs) in a controlled manner would further investigation of human neurogenesis and development of potential cell therapeutic applications to treat neurological diseases; however, generating such neural stem cells (NSCs) remains a challenge. In an attempt to characterize the cellular mechanisms involved in hESC differentiation into neuroprogenitor cells, we performed 2-DE using protein extracts from hESC-derived embryoid bodies (EBs) and neuroectodermal spheres (NESs) bearing neuroprogenitors. Of 47 differentially expressed protein spots, 28 nonredundant spots were shown to be upregulated in the NESs; these protein spots included neurogenesis-related proteins (TAF1, SEPT2, NPH3, and CRABP), as expected. Interestingly, 6 of these 28 protein spots were cytoskeleton-associated proteins (CSAP) such as Fascin-1, Cofilin-1, and Stathmin-1. Western-blot analyses confirmed the increased levels of these proteins in the NESs. Furthermore, immunostaining analysis showed that both Fascin-1 and Stathmin-1 were preferentially expressed in the inner rims of neural rosettes, which are characteristic features of neuroprogenitors in culture. We also confirmed prominent expression of Fascin-1 in (sub-)ventricular zone in E15.5 mouse fetal brain. Our results suggest that, in addition to the induction of those genes involved in neural development, hESC differentiation into the NES is associated with a marked reorganization of the cellular cytoskeleton.
Assuntos
Proteínas do Citoesqueleto/análise , Proteínas do Citoesqueleto/genética , Células-Tronco Embrionárias/citologia , Placa Neural/citologia , Neurogênese , Diferenciação Celular , Células Cultivadas , Proteínas do Citoesqueleto/metabolismo , Eletroforese em Gel Bidimensional , Células-Tronco Embrionárias/metabolismo , Humanos , Placa Neural/metabolismo , Proteoma/análise , Proteoma/genética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas em Tandem , Regulação para CimaRESUMO
The potential medical applications of animal cloning include xenotransplantation, but the complex molecular cascades that control porcine organ development are not fully understood. Still, it has become apparent that organs derived from cloned pigs may be suitable for transplantation into humans. In this study, we examined the pancreas of an adult cloned pig developed through somatic cell nuclear transfer (SCNT) using two-dimensional electrophoresis (2-DE) and Western blotting. Proteomic analysis revealed 69 differentially regulated proteins, including such apoptosis-related species as annexins, lamins, and heat shock proteins, which were unanimously upregulated in the SCNT sample. Among the downregulated proteins in SCNT pancreas were peroxiredoxins and catalase. Western blot results indicate that several antioxidant enzymes and the anti-apoptotic protein were downregulated in SCNT pancreas, whereas several caspases were upregulated. Together, these data suggest that the accumulation of reactive oxygen species (ROS) in the pancreas of an adult cloned pig leads to apoptosis.
Assuntos
Animais Geneticamente Modificados/genética , Clonagem de Organismos/métodos , Pâncreas/metabolismo , Proteoma/genética , Suínos/genética , Animais , Perfilação da Expressão GênicaRESUMO
The attenuation and immunoenhancing effects of rpoS and phoP Salmonella enterica serovar strain Typhi (Salmonella typhi) mutants have not been compared. Here, three S. typhi deletion mutants (phoP, rpoS, and rpoS-phoP double mutant) are constructed and these mutants are characterized with respect to invasiveness, virulence, and protective immune response compared with wild-type Ty2. It was found that phoP and phoP-rpoS deletion mutants are less invasive to HT-29 cells than the wild-type Ty2 and the rpoS single-deleted strain. The LD(50) of immunized mice was higher for phoP than for rpoS mutants, and the highest for the phoP-rpoS double mutant. In addition, all S. typhi mutants showed an increase in the specific serum IgG levels and T-cell-mediated immunity, and showed equal protection abilities against a wild-type Ty2 challenge after two rounds of immunization in BALB/c mice. It is concluded that phoP genes appear to play a more important role than rpoS genes in both cellular invasion and virulence of S. typhi, but not in immunogenicity in mice. Furthermore, the data indicate that the phoP-rpoS double mutant may show promise as a candidate for an attenuated typhoid vaccine.