Top PDF Gene expression and functional annotation of the human ciliary body epithelia.

Gene expression and functional annotation of the human ciliary body epithelia.

Gene expression and functional annotation of the human ciliary body epithelia.

Human Donor Eyes, Tissue Processing and Cell Sampling We selected seven eyes following previously published strict protocols [24]. We now selected adult donors as young possible, to avoid possible undiagnosed POAG [25]. Table S1 outlines the characteristics of the seven donor eyes. The medical histories revealed no prolonged disease state or medication use that could influence RNA expression or quality. Furthermore, the medical histories displayed neither glaucoma or other eye diseases nor any malignancies. Inspection of the eyes with light microscope revealed no abnormalities. Globes were enucleated 10–21 hours post mortem and were frozen within 24 hours total post mortem time. After removal of the cornea, the donor eyes were transferred to our department and visually inspected. Subsequently, they were snap frozen in isopentane in liquid nitrogen and stored at minus 80 uC. Next, twenty 20 m m cryosections of the CB were cut and mounted on slides covered with 1.0 mm PEN-membrane (PALM Carl Zeiss, MicroImaging GmbH, Munich, Germany). The slides were dehydrated and stained with Cresyl Violet (1 gram of Kresyl Violett (Merck, Frankfurt, Germany, art.5235), added to 1 L of 100% ethanol). Cresyl Violet staining was necessary to distinguish NPE. Next, the NPE and PE were separately isolated with a laser microdissection system (PALM Carl Zeiss, Microlaser Technologies AG, Germany) and captured in adhesive caps (AdhesiveCap 500 opaque, Carl Zeiss, Germany). Figure S1 shows the various steps.
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Derivation of human differential photoreceptor-like cells from the iris by defined combinations of CRX, RX and NEUROD.

Derivation of human differential photoreceptor-like cells from the iris by defined combinations of CRX, RX and NEUROD.

It has been hypothesized that retinal stem cells can be found in the ciliary body [14], postnatal retina, and the iris [15]. Pure populations of IPE cells isolated from rat and chicken irises were shown to demonstrate ‘‘stemness’’ [16]. A portion of purely isolated IPE cells of rodents, especially nestin-positive IPE cells, differentiated into multiple neuronal cell types, pan-neural marker- expressing cell types and retina-specific cell types without Figure 3. Induction of opsin genes in human iris-derived cells, ciliary epithelial cells and retina-derived cells by the retroviral infection of all the 6 genes and genes for RX , CRX and NEUROD . (A) RT-PCR analysis for genes of MAP2, rhodopsin, blue opsin and G3PDH in two kinds of iris cells after gene transfer of all the six genes. All six genes were infected into two kinds of iris cells: IPE and stromal cells derived from the peripheral iris, and purely isolated IPE cells. In both cell types, rhodopsin and blue opsin genes were up-regulated. ‘‘w/o’’: cultured iris-derived cells without gene transfer as a negative control. (B) Expression of the rhodopsin and blue opsin genes started two weeks after infection. ‘‘IS’’, ‘‘IPE’’ and ‘‘Iris (central)’’ indicate ‘‘iris-stromal cells’’, ‘‘iris pigment epithelial cells’’, and ‘‘central iris cells’’, respectively. ‘‘IS (GFP)’’ is ‘‘iris-stromal cells infected with the GFP gene’’. In all kinds of iris cells, transduction of the three genes, that are RX, CRX and NEUROD, enhanced expression of rhodopsin, blue opsin and green/red opsin. (C) RT-PCR analysis for genes for genes of neural crest-related markers in two kinds of iris-derived cells: iris stromal cells and iris pigmented epithelial cells. (D) Phase-contrast photomicrograph of ciliary epithelial cells from pars plana (left) and pars plicata (right). (E) RT- PCR analysis for genes of rhodopsin, blue opsin, green/red opsin and G3PDH in human ciliary epithelium (pars plicata and pars plana) after gene transfer of either all the six genes (SIX3, PAX6, RX, CRX, NEUROD, NRL) or three genes (CRX, RX and NEUROD). (F) RT-PCR analysis for the MAP2, rhodopsin, blue opsin, GFAP and G3PDH genes in retina-derived cells after transfer of all six genes. ‘‘w/o’’: Retina-derived cells without gene transfer 21 days after the start of cultivation. Genes for MAP2, rhodopsin, and blue opsin started to be expressed after the gene transfer.
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Functional annotation of rheumatoid arthritis and osteoarthritis associated genes by integrative genome-wide gene expression profiling analysis.

Functional annotation of rheumatoid arthritis and osteoarthritis associated genes by integrative genome-wide gene expression profiling analysis.

Rheumatoid arthritis (RA) is a common chronic systemic autoimmune disease that mainly affects the flexible joints. It is characterized by the inflammation of articular synovial. The lasting recurrent inflammation of synovial can lead to the deformation and destruction of cartilage and bones, which could result in disability of the patients [1,2]. RA mainly occurs in the 30,70 years old people and is more frequent in females than males. More than 1% of the world’s population may be affected by RA [3,4]. This disease brings great physiological and psychological burden to patients. However, the biological causes for RA remain largely unknown. Although infectious agents including viruses, bacteria and fungi have long been suspected, none has been comprehensively proved [5,6]. Previous researches have also investigated the potential associations between RA and environ- mental factors, such as smoking, vitamin D deficiency, etc [7,8]. It is now generally believed that the pathogenesis of RA is closely related to genetic factors. Certain genes such as the human leukocyte antigen (HLA). HLA-DR4 and DW4 antigen, were identified in more than 90% of the patients. These pathological factors are referred to as the RA-shared epitope [3,9].
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Mammary fat of breast cancer: gene expression profiling and functional characterization.

Mammary fat of breast cancer: gene expression profiling and functional characterization.

Mammary fat tissues were fixed in 10% formalin, processed and paraffin-embedded. Multiple sections (5 mm) were prepared and stained with hematoxylin and eosin for general morphological observation. For immunocytochemical staining, sections of fat tissues were incubated with anti-UCP1 antibodies (1:1000, Abcam, MA, USA) for 30 min at room temperature. Signals were detected using a biotinylated goat anti-rabbit secondary antibody (Vector Laboratories, CA, USA) in combination with the ABC kit (Vector Laboratories, CA, USA) and DAB substrate (Vector Laboratories, CA, USA). Samples were visualized using a Nikon Eclipse 80i upright microscope (Nikon, Tokyo, Japan). The intensity of the staining signal was measured and documented using Image-Pro Plus 6.0 image analysis software (Media Cybernetics, Inc. Silver Spring, MD, USA). The mean densitometry of the digital image was designated as representative UCP1 staining intensity (indicat- ing relative UCP1 expression). The signal density of tissue areas from five randomly selected images were counted in a blinded manner and subjected to statistical analysis.
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Influence of ERF3A/GSPT1 gene expression on susceptibility to carcinogenesis

Influence of ERF3A/GSPT1 gene expression on susceptibility to carcinogenesis

Despite gastric cancer incidence and mortality have decreased over the past decades, it is still the second most frequent cause of cancer-related death in the world (Crew & Neugut, 2006). Multiple genetic and epigenetic alterations in cell cycle regulators, cell adhesion molecules, DNA repair genes and telomerases, as well as genetic instability at microsatellite loci are the most common events implicated in the multistep process of gastric carcinogenesis (Tahara, 2004). However, gastric cancer is essentially a heterogeneous disease, and the events that underlie the malignant transformation of the gastric mucosa during the multistep process of gastric cancer pathogenesis remain uncertain (Zheng et al., 2006). Several histological classification for gastric cancer have been proposed, being the Lauren’s (1965) classification the most usual - two major forms of gastric tumors are distinguished according to their morphological and clinicopathological classifications: intestinal (well-differentiated) and diffuse (undifferentiated) type tumors. They differ in genetic susceptibility, pathologic profile, clinical presentation, and prognosis (Crew & Neugut, 2006; Cervantes et al., 2007). The intestinal type carcinoma is frequently preceded my multifocal atrophic gastritis and is more common in elderly man; the tumors show gland formation and are usually well demarcated. For the diffuse type tumors the precursor lesions are usually not identified and the prognosis is less favorable, they show no cohesiveness of tumors cells infiltrating the stroma and occurs preferably in women under 50 years old (Werner et al., 2001). Molecular pathology supports this classification by showing differences in the genetics pathways in the origin of both kinds of tumors (Werner et al., 2001; Tahara, 2004; Cervantes et al., 2007). Nevertheless, the prognosis for patients with advanced gastric cancer remains poor, much because gastric cancer is often diagnosed at an advanced stage. Therefore, it is still a challenge to detect stage-specific genetic abnormalities that may result in early diagnosis and aid in selecting therapy.
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Cloning and Functional Assessment of the Recombinant Human Hepcidin-25 in the Baculovirus Expression System

Cloning and Functional Assessment of the Recombinant Human Hepcidin-25 in the Baculovirus Expression System

The designed construct for the coding of human hepcidin-25 contained His-tag, TEV recognition site and coding sequence of human hepcidin immediately following ATG codon. The N- terminal His-tag was introduced to enable nickel- based purification. Recognition site for TEV was introduced so as to enable the removal of His-tag from the purified hepcidin-25. This strategy also allowed for recombinant hepcidin production with a minimum number of extra amino acids at its N- terminus. Hepcidin coding sequence has only one neutral extra amino acid (glycine). Because of its neutral nature, the added glycine is not considered to significantly affect the peptide folding and efficiency (Ladurner and Fersht 1997).
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The Impact of E-Commerce Securi ty, and National Environment  on Consumer adoption of Intern et Banking in Malaysia and  Singapore

The Impact of E-Commerce Securi ty, and National Environment on Consumer adoption of Intern et Banking in Malaysia and Singapore

Relative advantage is defined as the extent to which a person views an innovation as offering an advantage over previous ways of performing the same task (Roger, 1983; Agarwal & Prasad, 1997). Because Internet banking services allow customers to access their banking account from any location 24 hours a day and 7 days a week, it provides an enormous advantage and convenience to users (Tan & Teo, 2000). It also gives customers greater control over managing their finances, as they are able to check their accounts easily. Besides, a customer’s Internet experience, his or her banking needs can affect his adoption. As there are more financial products and services, it is expected that individuals with many financial accounts and who subscribe to many banking services will be more inclined to adopt Internet banking. Tan and Teo (2000) has reported that potential adopters of Internet banking services are likely to own multiple banking accounts and subscribe to various banking services. Rogers argues that potential adapters, who are allowed to experiment with an innovation will feel more comfortable with the innovation and are more likely to adopt it. Thus, if customers have the opportunity to try the innovation, certain fears of the unknown may be minimized. Government policy could also aid or hinder Internet diffusion (Mbarika, 2002). This is consistent with the national systems of innovation theory that posits that government policies may encourage or mandate technology development and adoption (King et. al., 1994; Wolcott et. al., 2001). Tan and Teo (2000) suggest that the greater the extent of government support for Internet commerce, the more likely Internet banking will be adopted, thus, confirming Goh’s (1995) suggestion that governments can play an interventionist and leading role in the diffusion of innovation. Potential users in turn would view new applications such as Internet banking services more favorably and hence be more like to use them. Thus, the second alternative hypothesis is:
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COL18A1 is highly expressed during human adipocyte differentiation

COL18A1 is highly expressed during human adipocyte differentiation

Collagen XVIII can generate two fragments, NC11-728 containing a frizzled motif which possibly acts in Wnt sig- naling and Endostatin, which is cleaved from the NC1 and is a potent inhibitor of angiogenesis. Collagen XVIII and Wnt signaling have recently been associated with adipogenic differentiation and obesity in some animal models, but not in humans. In the present report, we have shown that COL18A1 expression increases during human adipogenic dif- ferentiation. We also tested if polymorphisms in the Frizzled (c.1136C>T; Thr379Met) and Endostatin (c.4349G>A; Asp1437Asn) regions contribute towards susceptibility to obesity in patients with type 2 diabetes (113 obese, BMI ≥30; 232 non-obese, BMI < 30) of European ancestry. No evidence of association was observed between the allele c.4349G>A and obesity, but we observed a significantly higher frequency of homozygotes c.1136TT in obese (19.5%) than in non-obese individuals (10.9%) [P = 0.02; OR = 2.0 (95%CI: 1.07-3.73)], suggesting that the allele c.1136T is associated to obesity in a recessive model. This genotype, after controlling for cholesterol, LDL cholesterol, and triglycerides, was independently associated with obesity (P = 0.048), and increases the chance of obesity in 2.8 times. Therefore, our data suggest the involvement of collagen XVIII in human adipogenesis and susceptibility to obesity. Key words: COL18A1, obesity, adipogenesis, endostatin, frizzled.
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A gene wiki for community annotation of gene function.

A gene wiki for community annotation of gene function.

Participation in Gene Annotation To assess the effect of these gene stubs on Wikipedia, we first surveyed the edit logs for each of these pages that track contributions from the community (Figure 2A). For the 650 gene pages that were previously existing and amended in this effort, we found that the edit rates were roughly equal when comparing activity both before and after our automated efforts. Among the 7,500 new gene stubs created, edit rates were on average 10-fold less than for the pre-existing pages. However, due to the substantial difference in size of these groups, approximately 50% of all edits to gene pages were made on the newly created pages. These results demonstrate that in terms of the absolute number of edits, this effort roughly doubled the amount of mammalian gene annotation activity in Wikipedia. Given the relatively short period of time for which these entries have been available and the fact that this effort has not been previously announced in publication, we expect the rate of editing activity to continue to grow.
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Determinants and Consequences of   the Website Perceived Value

Determinants and Consequences of the Website Perceived Value

rich in value. Experiential value perceptions are based upon interactions involving either direct usage or distanced appreciation of goods and services. These interactions provide the basis for relativistic preferences held by the individual involved (Holbrook and Coffman, 1995). These preferences may be intrinsic_when the experience helps the shopper to realize some of his purposes_ or extrinsic when the experience is enjoyed for its own sake (Babin and Darden, 1995; Batra and Ahtola, 1991). Besides, they may be experienced as the consequence of active (the consumer assumes the role of co- producer) or reactive (that occurs when the individual simply apprehends, appreciate, or responds to an object) interactions with the product/service. The following experiential value matrix developed by Malhotra et al (2001) represents an integration of the intrinsic/extrinsic and active/reactive dimensions.
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Artificial induction of Sox21 regulates sensory cell formation in the embryonic chicken inner ear.

Artificial induction of Sox21 regulates sensory cell formation in the embryonic chicken inner ear.

Notch signalling throughout the developing chicken inner ear, and can be used to monitor reductions in endogenous levels of Notch activity (E. Chrysostomou, J. Gale and N. Daudet, in press). We focused our analysis on vestibular patches in which induction of Sox21 had clear effects on hair cell differentiation. We co-electroporated either the pTRE-FP635 or pTRE-FP635- Sox21 (these inducible constructs included the red fluorescent protein Turbo-FP635 instead of eGFP to identify induced cells) with the pT2K-Hes5::nd2eGFP reporter at E2, let the embryos develop to E10 stages, then dissected and cultured vestibular patches from three independent samples in the presence of Dox. After 12 hours of treatment, we found endogenous Notch activity (defined as Hes5::nd2eEGFP levels .500 above background in a 12-bits image) in 68.1% of control FP635-induced cells (n = 94/138), versus only 49% of FP635-Sox21 induced cells (n = 51/104; Fig. 6A,B). This difference was even greater after 48 hours: Notch activity was observed in an average of 64.5% of FP635 induced cells (n = 71/110), but in only 21.2% of FP635- Sox21 cells (n = 23/108; Fig. 6B). In some cases, a striking checkerboard-like pattern of FP635-Sox21 induced cells sur- rounded by strongly nd2EGFP-positive cells was observed (Fig. 6A, lower panels). We next quantified Hes5::nd2EGFP fluorescence levels of individual progenitor/support cells after 12 hours of Dox treatment. In control samples, there was no statistically significant difference in levels of Hes5::nd2EGFP fluorescence of FP635-induced versus non-induced cells (Mann Whitney U test; p = 0.000; total n nuclei = 180 transfected/432 untransfected in n = 3 samples). However, there was a significant decrease (Mann-Whitney U test = 11232; p = 0.000) in the levels of nd2EGFP expression in FP635-Sox21 transfected cells (n = 178; n = 3 samples) when compared to FP635-transfected cells (n = 180; n = 3 samples) (Fig. 6C). On the other hand, expression of the Notch ligands Delta1 and Serrate1 did not appear to differ between control and Sox21-induced cells at this time point (Fig. 7A–F), suggesting that the reduction in
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Transcript annotation in FANTOM3: mouse gene catalog based on physical cDNAs.

Transcript annotation in FANTOM3: mouse gene catalog based on physical cDNAs.

N. Maeda, M. Itoh, K. Shibata, J. Kawai, P. Carninci, and Y. Hayashizaki are in the Genome Science Laboratory, Discovery Research Institute, RIKEN Wako Institute, Wako, Japan. T. Kasukawa, R. Oyama, J. Gough, M. Frith, M. Kanamori-Katayama, S. Katayama, T. Kawashima, C. Kai, J. Kawai, P. Carninci, and Y. Hayashizaki are in the Genome Exploration Research Group (Genome Network Project Core Group), RIKEN Genomic Sciences Center, RIKEN Yokohama Institute, Yokohama, Japan. T. Kasukawa is in the Broadband Communication Service Business Unit, Network Service Solution Business Group, NTT Software Corporation, Yokohama, Japan. M. Frith, R. N. Aturaliya, and R. D. Teasdale are at the Australian Research Council Center in Bioinformatics, Institute for Molecular Bioscience, University of Queensland, St Lucia, Queensland, Australia. P. G. Engstro¨m and B. Lenhard are in the Computational Biology Unit, Bergen Center for Computational Science, University of Bergen, Bergen, Norway, and the Programme for Genomics and Bioinformatics, Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden. S. Batalov and C. F. Fletcher are at the Genomics Institute of the Novartis Research Foundation, San Diego, California, United States of America. K. W. Beisel is in the Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska, United States of America. C. J. Bult, M. Furuno, D. Hill, and Y. Zhu are in the Mouse Genome Informatics Consortium, The Jackson Laboratory, Bar Harbor, Maine, United States of America. A. R. R. Forrest, T. Ravasi, and D. A. Hume are at the Australian Research Council Special Research Centre for Functional and Applied Genomics, Institute for Molecular Bioscience, University of Queensland, Brisbane, Queensland, Australia. M. Katoh is in the Genetics and Cell Biology Section, National Cancer Center Research Institute, Tokyo, Japan. J. Quackenbush is at the Institute for Genomic Research, Rockville, Maryland, United States of America. B. Z. Ring is at Applied Genomics, Sunnyvale, California, United States of America. K. Sugiura is at The Jackson Laboratory, Bar Harbor, Maine, United States of America. Y. Takenaka is in the Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, Osaka, Japan. C. A. Wells is in the School of Biomolecular and Biomedical Science, Eskitis Institute for Cell and Molecular Therapies, Griffith University, Nathan, Queensland, Australia. Y. Hayashizaki is at Yokohama City University, Yokohama, Japan, and in the Graduate School of Comprehensive Human Science, University of Tsukuba, Tsukuba, Japan. * To whom correspondence should be addressed. E-mail: rgscerg@gsc.riken.jp ¤a Current address: Functional Genomics Subunit, Center for Developmental Biology, RIKEN, Kobe, Japan
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Buying Behavior Of Organic Vegetables Product The Effects Of Perceptions Of Quality And Risk

Buying Behavior Of Organic Vegetables Product The Effects Of Perceptions Of Quality And Risk

quality of alternatives with regard to price within a category (Jin & Suh, 2005). Organic vegetable products have advantages and technologies related of environmental friendly. Perceived quality is not the actual quality of the brands or products. Rather, it is the consumers’ judgment about an entity’s or a service’s overall excellence or superiority (Aaker, 1991). Sometimes is directly related to the reputation of the firm that manufactures the product (Davis et al. 2003), and viewed as the degree and direction of discrepancy between consumers’ perceptions and expectations (Chen & Chang, 2005). Perceived quality and perception of quality had closer theoretical, perception defined is the mental process that persons go through in selecting, organizing and interpreting information into meaningful patterns (Truong & Yap, 2010:532). It can be interpreted that perception of quality is overall judgment of superior quality of organic products as result from selecting, organizing and interpreting form the alternative product. Measurement of customer perception of quality on organic products is divide on several things, included guarantee (origin, brand, label, variety), organoleptic characteristic (firmness, color, flavor, aroma), and external factors (damage, size, price) (Carrasco et al., 2012:1422). In other side on organic product it measured with environmental concern, environmental consideration, environmental performance, environmental image, and environmental reputation (Chen & Chang, 2013:71).
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Gene expression during the generation and activation of mouse neutrophils: implication of novel functional and regulatory pathways.

Gene expression during the generation and activation of mouse neutrophils: implication of novel functional and regulatory pathways.

As part of the Immunological Genome Project (ImmGen), gene expression was determined in unstimulated (circulating) mouse neutrophils and three populations of neutrophils activated in vivo, with comparison among these populations and to other leukocytes. Activation conditions included serum-transfer arthritis (mediated by immune complexes), thioglycollate-induced peritonitis, and uric acid-induced peritonitis. Neutrophils expressed fewer genes than any other leukocyte population studied in ImmGen, and down-regulation of genes related to translation was particularly striking. However, genes with expression relatively specific to neutrophils were also identified, particularly three genes of unknown function: Stfa2l1, Mrgpr2a and Mrgpr2b. Comparison of genes up-regulated in activated neutrophils led to several novel findings: increased expression of genes related to synthesis and use of glutathione and of genes related to uptake and metabolism of modified lipoproteins, particularly in neutrophils elicited by thioglycollate; increased expression of genes for transcription factors in the Nr4a family, only in neutrophils elicited by serum-transfer arthritis; and increased expression of genes important in synthesis of prostaglandins and response to leukotrienes, particularly in neutrophils elicited by uric acid. Up-regulation of genes related to apoptosis, response to microbial products, NFkB family members and their regulators, and MHC class II expression was also seen, in agreement with previous studies. A regulatory model developed from the ImmGen data was used to infer regulatory genes involved in the changes in gene expression during neutrophil activation. Among 64, mostly novel, regulatory genes predicted to influence these changes in gene expression, Irf5 was shown to be important for optimal secretion of IL-10, IP-10, MIP-1a, MIP-1b, and TNF-a by mouse neutrophils in vitro after stimulation through TLR9. This data-set and its analysis using the ImmGen regulatory model provide a basis for additional hypothesis-based research on the importance of changes in gene expression in neutrophils in different conditions.
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Clinics  vol.67 número2

Clinics vol.67 número2

This study evaluated the effects of c-tocotrienol in the modulation of cellular aging in HDFs. Our results demon- strated clear changes in cellular morphology, decreased cellular proliferation, and increased senescence-associated b-galactosidase activity in senescent HDFs. Cellular and organism morphological changes are typical features of a Figure 3 - Senescence-associated b-galactosidase (SA b-gal) staining in young (a), pre-senescent (b), and senescent HDFs (c). Positive blue stains of SA b-gal appeared in senescent HDFs as indicated by arrows. Micrographs are shown at 100X magnification (A). Quantitative analysis of positive b-galactosidase-stained cells in HDFs during cellular aging. The percentage of cells positive for SA-b-gal staining was significantly increased in senescent cells. The incubation of senescent cells with c-tocotrienol (70 mM for 24 h) did not produce significant changes in the percentage of SA-b-gal-positive cells. a Denotes p,0.05 compared to untreated young HDFs, b
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The CCAAT/enhancer-binding protein beta-2 isoform (CEBPβ-2) upregulates galectin-7 expression in human breast cancer cells.

The CCAAT/enhancer-binding protein beta-2 isoform (CEBPβ-2) upregulates galectin-7 expression in human breast cancer cells.

receptors [26–28]. An increase of C/EBPb mRNA also correlates with metastatic breast cancer and high tumor grade, making it an interesting biomarker for subsets of tumors with a poor prognosis [28–30]. Moreover, our preliminary results have shown that transfection of C/EBPb-2 in MCF-7 cells increases their motility (Figure S1). Similar results were obtained when we increased galectin-7 expression (Figure S2), consistent with our previous findings that high levels of galectin-7 increase metastasis of breast cancer cells to the bone and the lung [9]. In fact, our results may have implications in other types of cancer where gal-7 is expressed, most notably in transformed keratinocytes. Both gal-7 and C/ EBPb-2 have been shown to be involved in the differentiation of keratinocytes [31]. Whether suppression of C/EBPb-2 will necessarily reduces gal-7 is currently unknown but likely since previous studies showing that retinoic acid, which suppresses the expression of C/EBPb-2 target genes in keratinocytes [32–33], also inhibits gal-7 expression [34]. Interestingly, C/EBPb-2 has been shown to be expressed at high levels in most mammary tumors and is the most frequent isoform found in human breast cancer cell lines [35]. Its expression in MCF10A cells has also been shown to induce in vitro epithelial to mesenchymal transition associated with increased invasive properties [17]. However, while C/EBPb-2 is generally considered the most transcriptionally active of all three C/EBPb isoforms, we cannot completely rule out the implication of other isoforms in regulating gal-7 expression. The C/EBPb-3 isoform, for instance, is expressed at high levels in some breast carcinoma, most notably in basal-like breast cancer [36– 37]. Future studies with isoform-specific antibodies will thus be needed to correlate the expression of gal-7 with specific C/EBPb isoforms in breast cancer tissues. It is important to note, however, that other transcription factors can also upregulate gal-7 and may thus compensate for suppression of C/EBPb-2. This is particularly true for breast cancer cells which often express a mutant form of p53, which is capable of inducing gal-7 [38]. Future investigations will thus be needed to determine whether direct suppression of galectin-7 expression or via specific targeting C/EBPb are valuable alternatives to inhibit breast cancer progression.
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Efeito da suplementação da vitamina E (a-tocoferol) ou D3 na expressão gênica de TLR4 e adipocinas. Estudos in vivo e in vitro

Efeito da suplementação da vitamina E (a-tocoferol) ou D3 na expressão gênica de TLR4 e adipocinas. Estudos in vivo e in vitro

Production of IL-6 increases with adiposity, and circu- lating IL-6 concentrations are highly correlated with percentage of body fat and insulin resistance [33]. Vita- min E supplementation in patients with diabetes decreases the levels of proinflammatory cytokines, such as IL-1, TNF-a, IL-6, and reactive C protein, in serum and stimulated monocytes [34,35]. In addition, high intake of foods rich in vitamin E demonstrated an inverse correlation with levels of C reactive protein and IL-6 [36]. Elevated TNF-a concentrations cause stress oxidation in monocytes which, conversely, increases low density lipoprotein (LDL) oxidation [37]. Oxidative modification of lipoproteins is an essential component of the atherogenic process [34]. Manning et al. (2004) showed that vitamin E supplementation decreased plasma peroxide concentration in obese individuals [26]. In the present study, we demonstrated that a vitamin D3- and vitamin E-supplemented diet decreases IL-6 levels in adipose tissue and in 3T3-L1 adipocytes. Taken together, these results demonstrate that vitamin E and vitamin D3 could have a direct effect on adipocytes.
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Expression of the human glucokinase gene: important roles of the 5' flanking and intron 1 sequences.

Expression of the human glucokinase gene: important roles of the 5' flanking and intron 1 sequences.

Since only 4 kb of liver-specific exon 5 9 flanking sequence and intron 1 sequences are conserved among mammals, and DNase I hypersensitive sites have been mapped to both of these regions [7,21,22] we tested the ability of these sequences to regulate luciferase reporter gene expression in two human liver cell lines. In addition to testing the reporter genes in the well-established HepG2 cell line that has been used in a number of experiments studying the expression of the glucokinase gene promoter [13,20,32–34], we also tested the constructs in a second human liver cell line, the normal liver cell line L-02 [27]. To examine whether the L-02 cells would be suitable for studying the regulation of glucokinase gene expression we first determined whether L-02 and HepG2 cells express similar profiles of metabolic genes. Expression of GCK, GCKR, components of the insulin signaling pathway (INSR, IRS1, and IRS2), other metabolic enzymes (IPK, PCK1, PYGL, GYS2, and FASN), as well as selected DNA binding proteins involved in the regulation of metabolism (ChREBP, SREBF, NHF4A, PPARG, and FOXO1) were assessed by RT-PCR for cells grown in culture in the presence or absence of insulin or fetal calf serum (Fig. S3). As is typically seen for hepatic cell lines [5], significant expression of the glucokinase gene was not detected in either L-02 or HepG2 cells. Comparing the gene expression profiles of L-02 to HepG2 showed that both cell lines usually express similar levels for most of the genes examined, although slightly higher levels of GCKR and lower levels of SREBF were detected in L-02 (Fig. S3). The increased level of GCRK expression observed by RT-PCR in L-02 cells was confirmed by quantitative real-time RT-PCR (results not shown). RT-PCR experiments showed that there was little change in the expression of the genes due to the presence or absence of insulin or serum for either HepG2 or L-02 cells, except for GCKR in HepG2 (confirmed by quantitative real time RT-PCR, results not shown). It appears that L-02 should be an appropriate cell model system for understanding liver-specific expression of the human glucokinase gene.
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The influence of the arc plasma treatment on the structure and microhardness C120U carbon tool steel

The influence of the arc plasma treatment on the structure and microhardness C120U carbon tool steel

After arc plasma electric treatment on the cross section of the surface layer was distinguished two areas with the different microstructure were observed in respect to the parent material. The first area – remelted zone (RZ) and second area – heat affected zone (HAZ). Optical microscopic images of a single track are presented on Fig. 2. The influence of current intensity of the electric arc plasma on the dimensions of RZ and HAZ are shown in Table 2. With increased current intensity (for fixed speed rate of source = 0,2 m/min) the depth of the RZ increased too.
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Influence of tungsten and titanium on the structure of chromium cast iron

Influence of tungsten and titanium on the structure of chromium cast iron

Studies have proved that structure of the chromium cast iron greatly depends on the additionally introduced elements such as titanium and tungsten. Titanium is a carbide-forming element, but in contrast to other elements of this type it does not form complex carbides in the chromium cast iron, but only a TiC carbide, which is formed at high temperature in the liquid metal. Tungsten is also a carbide-forming (and pearlite-forming) element but, like titanium, is rarely used in the manufacture of chromium cast iron. High melting points of tungsten and titanium may cause difficulties in the metallurgical process of chromium cast iron manufacture. Tungsten effect on the mechanical properties is similar to that of molybdenum, although it is weaker. Tungsten increases the hardenability of cast iron. Currently, the use of tungsten can be justified because of its price slightly lower than that of molybdenum.
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