Wilms' tumor is associated with mutations of WT1, a zinc-finger transcription factor that is essential for the development of the metanephric kidney and the urogenital system. High levels of WT1 ...expression also have been detected in myeloid leukemia cells, suggesting that WT1 may be important in other neoplasms as well. To seek a role of high level expression of WT1 in the differential arrest characteristic of myeloid leukemia, WT1 or its zinc-finger domain alone was stably expressed in human promyeloid leukemia (HL-60) cells and the ability of 12-O-tetradecanoyl-phorbol-13-acetate (TPA) to induce macrophage differentiation was examined. HL-60 cell differentiation was completely arrested in TPA treated cells that expressed WT1 or its zinc-finger domain alone whereas TPA fully induced macrophage differentiation in control HL-60 cells, indicating that high level expression of WT1 is capable of differentiation arrest of myeloid cells and that its effect may be mediated through its zinc-finger domain. To determine if the zinc-finger domain of WT1 directly influences transcription, it was brought to promoter DNA as a fusion protein with the Gal4 DNA binding domain. The fusion protein failed to regulate transcription of a reporter gene but when the zinc-finger domain of WT1 was brought to DNA with a promoter containing two upstream WT1-binding sites, reporter gene expression was activated approximately threefold, suggesting that WT1 interferes with myeloid differentiation through the ability of its zinc-finger domain to compete with other transcription factors for common promoter elements.
MicroRNAs (miRNAs) can post-transcriptionally regulate gene expression and have been shown to be critical regulators to the fine-tuning of epithelial immune responses. However, the role of miRNAs in ...bovine responses to E. coli and S. aureus, two mastitis causing pathogens, is not well understood.
The global expression of miRNAs in bovine mammary epithelial cells (MAC-T cells) challenged with and without heat-inactivated Staphylococcus aureus (S. aureus) or Escherichia coli (E. coli) bacteria at 0, 6, 12, 24, and 48 hr was profiled using RNA-Seq. A total of 231 known bovine miRNAs were identified with more than 10 counts per million in at least one of 13 libraries and 5 miRNAs including bta-miR-21-5p, miR-27b, miR-22-3p, miR-184 and let-7f represented more than 50% of the abundance. One hundred and thirteen novel miRNAs were also identified and more than one third of them belong to the bta-miR-2284 family. Seventeen miRNAs were significantly (P < 0.05) differentially regulated by the presence of pathogens. E. coli initiated an earlier regulation of miRNAs (6 miRNAs differentially regulated within the first 6 hrs post challenge as compared to 1 miRNA for S. aureus) while S. aureus presented a delayed response. Five differentially expressed miRNAs (bta-miR-184, miR-24-3p, miR-148, miR-486 and let-7a-5p) were unique to E. coli while four (bta-miR-2339, miR-499, miR-23a and miR-99b) were unique to S. aureus. In addition, our study revealed a temporal differential regulation of five miRNAs (bta-miR-193a-3p, miR-423-5p, miR-30b-5p, miR-29c and miR-un116) in unchallenged cells. Target gene predictions of pathogen differentially expressed miRNAs indicate a significant enrichment in gene ontology functional categories in development/cellular processes, biological regulation as well as cell growth and death. Furthermore, target genes were significantly enriched in several KEGG pathways including immune system, signal transduction, cellular process, nervous system, development and human diseases.
Using next-generation sequencing, our study identified a pathogen directed differential regulation of miRNAs in MAC-T cells with roles in immunity and development. Our study provides a further confirmation of the involvement of mammary epithelia cells in contributing to the immune response to infecting pathogens and suggests the potential of miRNAs to serve as biomarkers for diagnosis and development of control measures.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
An abstract of an article by Guan et al on determining the effect of maternal nutrition management during mid-gestation on the skeletal muscle transcriptome in progeny using 3 dietary treatments is ...presented. Results indicate the skeletal muscle transcriptome and associated metabolic functions prior to slaughter are affected by mid-gestation maternal plane of nutrition and may be regulated by epigenetic factors.
We retrospectively evaluated 18fluoro-2-deoxyglucose positron emission tomography (FDG-PET) scans in 172 patients with lymphoma and correlated results with pathologic diagnosis using the World Health ...Organization (WHO) classification system. In total, FDG-PET detected disease in at least one site in 161 patients (94%) and failed to detect disease in 11 patients (6%). The most frequent lymphoma diagnoses were diffuse large B-cell lymphoma (LBCL; n = 51), Hodgkin lymphoma (HL; n = 47), follicular lymphoma (FL; n = 42), marginal zone lymphoma (MZL; n = 12), mantle cell lymphoma (MCL; n = 7), and peripheral T-cell lymphoma (PTCL; n = 5). FDG-PET detected disease in 100% of patients with LBCL and MCL and in 98% of patients with HL and FL. In contrast, FDG-PET detected disease in only 67% of MZL and 40% of PTCL. Comparison with bone marrow biopsies showed that FDG-PET was not reliable for detection of bone marrow involvement in any lymphoma subtype.
A rat brain cDNA library was screened by using a mixture of oligonucleotides whose sequences were deduced from the amino acid sequence of a human placental protein-tyrosine-phosphatase (PTPase; EC ...3.1.3.48) reported by Charbonneau et al. Charbonneau, H., Tonks, N. K., Walsh, K. A. \& Fischer, E. H. (1988) Proc. Natl. Acad. Sci. USA 85, 7182-7186. The isolated clones encode a PTPase of 432 amino acids having a mass of 49,679 daltons and showing 97% sequence identity to the corresponding 321 residues of the placental enzyme. The coding sequence of the PTPase was placed behind a bacteriophage T7 promoter and the protein was expressed in Escherichia coli. The recombinant protein had a molecular weight of 50,000 by SDS/PAGE analysis and showed an absolute specificity for phosphotyrosine-containing substrates. Northern analysis documented that there were two sizes of RNA, 4.3 and 2.0 kilobases, which encode the PTPase. Both transcripts were present in a number of tissues, and the smaller RNA appears to arise by the use of an alternative polyadenylylation signal. The PTPase was also localized by in situ hybridization in the rat central nervous system. A diffuse pattern of hybridization signal is seen in a number of brain areas, with the hippocampus showing the highest levels of mRNA. Sequences located at the C terminus of the rat brain PTPase contain possible sites for phosphorylation as well as signals which could serve for membrane attachment.
By performing studies using the kinetic Monte Carlo method, we have simulated the energetic growth process in pulsed laser deposition. In our model, we focus on the influence of particles' kinetic ...energies on island aggregation and obtain the different results in the cases of the average kinetic energy
E
k=0, 5 and 10 eV. The results indicate that in the early stage of film growth, the nucleation density still obeys the general scaling function that is different from the ordinary power law form. Moreover, we analyze the simulation results and find a similar effect of incident particles' kinetic energies and substrate temperature on film deposition, which is in agreement with other reported experimental and theoretical results.
Abstract
Volatile fatty acids (VFAs), the end products of rumen fermentation, are the principal energy source for ruminants which potentially affect carcass yield and fat deposition in beef cattle. ...Rumen microbiota affects lipid metabolism in ruminants which might contribute to the variation in fat deposition. In this study, rumen fluid samples were collected from 204 beef steers raised under feedlot in Roy Berg Kinsella Research Ranch, University of Alberta. Individual VFA concentrations were measured using gas chromatography, and the bacterial and archaeal abundances were estimated by measuring total copy numbers of their respective 16S rRNA genes using qPCR. Animal phenotypic measures including body weight (BW), dry matter intake (DMI), average daily gain (ADG), hot carcass weight (HCW), age at slaughter (age), ribeye area (CREA), adipose fat thickness (AFAT), lean meat yield (LMY) and marbling score (MS) were collected. The relationships among microbiota, VFAs and carcass traits were evaluated using stepwise, backward linear regression with a significant level of 0.05. The HCW was positively correlated with BW and ADG and was negatively correlated with propionate. Negative correlations were identified between age and BW, RFI, acetate, propionate, and butyrate. CREA had positive correlations with BW and DMI, and negative correlations with acetate. AFAT was positively correlated with DMI, BW and propionate while, acetate and butyrate tentatively positively correlated (P < 0.1) with AFAT. The LMY was negatively correlated with BW, DMI, acetate, propionate and butyrate. Marbling score was positively correlated with DMI, acetate, propionate and butyrate. The relationship between rumen microbial abundance and the above carcass and meat quality-related traits were not detected in this study. Our preliminary results reveal that rumen VFAs may contribute to carcass and meat quality traits in beef cattle, suggesting the potential application to improving carcass and meat quality in beef steers through manipulating rumen VFA.
PDF
