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Hung-Fu Liao, Chu-Fan Mo, Shinn-Chih Wu, Dai-Han Cheng, Chih-Yun Yu, Kai-Wei Chang, Tzu-Hao Kao, Chia-Wei Lu, Marina Pinskaya, Antonin Morillon, Shih-Shun Lin, Winston T K Cheng, Déborah Bourc'his, Timothy Bestor, Li-Ying Sung, Shau-Ping Lin
Nuclear transfer (NT) is a technique used to investigate the development and reprogramming potential of a single cell. DNA methyltransferase-3-like, which has been characterized as a repressive transcriptional regulator, is expressed in naturally fertilized egg and morula/blastocyst at pre-implantation stages. In this study, we demonstrate that the use of Dnmt3l-knockout (Dnmt3l-KO) donor cells in combination with Trichostatin A treatment improved the developmental efficiency and quality of the cloned embryos...
October 2015: Reproduction: the Official Journal of the Society for the Study of Fertility
Haitao Li, Feng Zhu, Lisa A Boardman, Lei Wang, Naomi Oi, Kangdong Liu, Xiang Li, Yang Fu, Paul J Limburg, Ann M Bode, Zigang Dong
BACKGROUND: Aspirin intake reduces the risk of colorectal cancer (CRC), but the molecular underpinnings remain elusive. Epidermal growth factor receptor (EGFR), which is overexpressed in about 80% of CRC cases, is implicated in the etiology of CRC. Here, we investigated whether aspirin can prevent CRC by normalizing EGFR expression. METHODS: Immunohistochemistry staining was performed on paraffin-embedded tissue sections from normal colon mucosa, adenomatous polyps from FAP patients who were classified as regular aspirin users or nonusers...
May 1, 2015: EBioMedicine
Andrew J O'Brien, Linda A Villani, Lindsay A Broadfield, Vanessa P Houde, Sandra Galic, Giovanni Blandino, Bruce E Kemp, Theodoros Tsakiridis, Paola Muti, Gregory R Steinberg
Aspirin, the pro-drug of salicylate, is associated with reduced incidence of death from cancers of the colon, lung and prostate and is commonly prescribed in combination with metformin in individuals with type 2 diabetes. Salicylate activates the AMP-activated protein kinase (AMPK) by binding at the A-769662 drug binding site on the AMPK β1-subunit, a mechanism that is distinct from metformin which disrupts the adenylate charge of the cell. A hallmark of many cancers is high rates of fatty acid synthesis and AMPK inhibits this pathway through phosphorylation of acetyl-CoA carboxylase (ACC)...
July 15, 2015: Biochemical Journal
Sarah McKenna, Clyde J Wright
Multiple mediators of septic shock are regulated by the transcription factor nuclear factor κB (NFκB). However, complete NFκB inhibition can exacerbate disease, necessitating evaluation of targeted strategies to attenuate the pro-inflammatory response. Here, we demonstrate that in murine macrophages, low-dose NFκB inhibitors specifically attenuates lipopolysaccharide (LPS)-induced IκBβ degradation and the expression of a select subset of target genes (encoding IL1β, IL6, IL12β). Gain- and loss-of-function experiments demonstrate the necessary and sufficient role of inhibitor of NFκB family member IκBβ (also known as NFKBIB) in the expression of these genes...
June 1, 2015: Journal of Cell Science
Wen-ping Zhou, Hui Zhang, Yu-xia Zhao, Gang-qiong Liu, Jin-ying Zhang
OBJECTIVE: Myocyte enhancer factor-2A (MEF 2A) has been shown to be involved in atherosclerotic lesion development, but its role in preexisting lesions is still unclear. In the present study we aim to assess the role of MEF 2A in the progression of pre-existing atherosclerosis. METHODS: Eighty apolipoprotein E-deficient mice (APOE KO) were randomly allocated to control, scramble and MEF 2A RNA interference (RNAi) groups, and constrictive collars were used to induce plaque formation...
2015: PloS One
Scott T Chancey, Sonia Agrawal, Max R Schroeder, Monica M Farley, Hervé Tettelin, David S Stephens
Macrolide resistance in Streptococcus pneumoniae emerged in the U.S. and globally during the early 1990's. The RNA methylase encoded by erm(B) and the macrolide efflux genes mef(E) and mel were identified as the resistance determining factors. These genes are disseminated in the pneumococcus on mobile, often chimeric elements consisting of multiple smaller elements. To better understand the variety of elements encoding macrolide resistance and how they have evolved in the pre- and post-conjugate vaccine eras, the genomes of 121 invasive and ten carriage isolates from Atlanta from 1994 to 2011 were analyzed for mobile elements involved in the dissemination of macrolide resistance...
2015: Frontiers in Microbiology
Sara Pensa, Marco Demaria, Lidia Avalle, Isaia Barbieri, Annalisa Camporeale, Valeria Poli
UNLABELLED: Abstract Background: The pro-oncogenic transcription factor STAT3 is constitutively active in tumours of many different origins, which often become addicted to its activity. STAT3 is believed to contribute to the initial survival of pre-cancerous cells as well as to hyper-proliferation and, later, metastasis. MATERIALS AND METHODS: To evaluate the contribution of enhanced STAT3 activation in a controlled model system, we generated knock-in mice in which a mutant constitutively active Stat3C allele replaces the endogenous wild-type allele and analysed its contribution to breast tumorigenesis...
June 2012: Hormone Molecular Biology and Clinical Investigation
Himanshu Sharma, Michelle A Digman, Natasha Felsinger, Enrico Gratton, Michelle Khine
We introduce a manufacturable and scalable method for creating tunable wrinkled ferromagnetic-metallic structures to enhance fluorescence signals. Thin layers of nickel (Ni) and gold (Au) were deposited onto a pre-stressed thermoplastic (shrink wrap film) polymer. Heating briefly forced the metal films to buckle when the thermoplastic retracted, resulting in multi-scale composite 'wrinkles'. This is the first demonstration of leveraging the plasmons in such hybrid nanostructures by metal enhanced fluorescence (MEF) in the near-infrared wavelengths...
2014: Optical Materials Express
Heather L Armata, Sally Chamberland, Lauren Watts, Hwi Jin Ko, Yongjin Lee, Dae Young Jung, Jason K Kim, Hayla K Sluss
Diabetes is a growing health care issue, and prediabetes has been established as a risk factor for type 2 diabetes. Prediabetes is characterized by deregulated glucose control, and elucidating pathways which govern this process is critical. We have identified the wild-type (WT) p53-inducible phosphatase (WIP1) phosphatase as a regulator of glucose homeostasis. Initial characterization of insulin signaling in WIP1 knockout (WIP1(KO)) murine embryo fibroblasts demonstrated reduced insulin-mediated Ak mouse transforming activation...
January 2015: Molecular Endocrinology
Eugenio Barone, Sebastien Mosser, Patrick C Fraering
Rapid remodeling of the actin cytoskeleton in the pre- and/or post-synaptic compartments is responsible for the regulation of neuronal plasticity,which is an important process for learning and memory. Cofilin1 plays an essential role in these processes and a dysregulation of its activity was associated with the cognitive decline observed during normal aging and Alzheimer's disease (AD). To understand the mechanism(s) regulating Cofilin1 activity we evaluated changes occurring with regard to Cofilin1 and its up-stream regulators Lim kinase-1 (LIMK1) and Slingshot phosphatase-1 (SSH1) in (i) human AD brain, (ii) 1-, 4-, and 10-months old APP/PS1 mice, (iii) wildtype 3-, 8-, 12-, 18- and 26-months old mice, as well as in cellular models including (iv) mouse primary cortical neurons (PCNs, cultured for 5, 10, 15 and 20 days in vitro) and (v) mouse embryonic fibroblasts (MEF)...
December 2014: Biochimica et Biophysica Acta
Anatoliy Dragan, Chris D Geddes
We present a potentially highly sensitive and selective bio-assay for the potential detection of any five different DNA sequences from one sample in one well. The assay is based on a DNA "rapid catch and signal" (DNA-RCS) technology developed for the detection of different DNA sequences from a sample well area. Our signal amplification utilizes the metal-enhanced fluorescence (MEF) of dyes attached to the probe-DNAs, which hybridizes with the pre-formed mixture of anchor-DNA scaffolds on silver island films (SiFs)...
November 2014: Journal of Fluorescence
Soham Chanda, Cheen Euong Ang, Jonathan Davila, ChangHui Pak, Moritz Mall, Qian Yi Lee, Henrik Ahlenius, Seung Woo Jung, Thomas C Südhof, Marius Wernig
Direct conversion of nonneural cells to functional neurons holds great promise for neurological disease modeling and regenerative medicine. We previously reported rapid reprogramming of mouse embryonic fibroblasts (MEFs) into mature induced neuronal (iN) cells by forced expression of three transcription factors: ASCL1, MYT1L, and BRN2. Here, we show that ASCL1 alone is sufficient to generate functional iN cells from mouse and human fibroblasts and embryonic stem cells, indicating that ASCL1 is the key driver of iN cell reprogramming in different cell contexts and that the role of MYT1L and BRN2 is primarily to enhance the neuronal maturation process...
August 12, 2014: Stem Cell Reports
Sang-Bing Ong, Andrew R Hall, Rachel K Dongworth, Siavash Kalkhoran, Aswin Pyakurel, Luca Scorrano, Derek J Hausenloy
The mechanism through which the protein kinase Akt (also called PKB), protects the heart against acute ischaemia-reperfusion injury (IRI) is not clear. Here, we investigate whether Akt mediates its cardioprotective effect by modulating mitochondrial morphology. Transfection of HL-1 cardiac cells with constitutively active Akt (caAkt) changed mitochondrial morphology as evidenced by an increase in the proportion of cells displaying predominantly elongated mitochondria (73 ± 5.0 % caAkt vs 49 ± 5.8 % control: N=80 cells/group; p< 0...
March 2015: Thrombosis and Haemostasis
Jelle Praet, Eva Santermans, Jasmijn Daans, Debbie Le Blon, Chloé Hoornaert, Herman Goossens, Niel Hens, Annemie Van der Linden, Zwi Berneman, Peter Ponsaerts
While multiple rodent preclinical studies, and to a lesser extent human clinical trials, claim the feasibility, safety, and potential clinical benefit of cell grafting in the central nervous system (CNS), currently only little convincing knowledge exists regarding the actual fate of the grafted cells and their effect on the surrounding environment (or vice versa). Our preceding studies already indicated that only a minor fraction of the initially grafted cell population survives the grafting process, while the surviving cell population becomes invaded by highly activated microglia/macrophages and surrounded by reactive astrogliosis...
2015: Cell Transplantation
Hong-Lin He, Ying-En Lee, Yow-Ling Shiue, Sung-Wei Lee, Li-Ching Lin, Tzu-Ju Chen, Ting-Feng Wu, Chung-Hsi Hsing, Hsuan-Ying Huang, Jaw-Yuan Wang, Chien-Feng Li
BACKGROUND AND OBJECTIVES: Neoadjuvant concurrent chemoradiotherapy (CCRT) followed by surgery is the standard treatment for locally advanced rectal cancer. Through data mining from published transcriptomic database, we identified Regenerating Gene Type IV (REG4) as the most significantly associated gene with resistance to CCRT. This study examined the prognostic impact of REG4 expression in patients with rectal cancer receiving neoadjuvant CCRT. METHODS: REG4 immunohistochemistry was retrospectively assessed for pre-treatment biopsy specimens from 172 rectal cancer patients who received neoadjuvant CCRT followed by surgery without initial distant metastasis...
December 2014: Journal of Surgical Oncology
Annalisa Camporeale, Marco Demaria, Emmanuelle Monteleone, Carlotta Giorgi, Mariusz R Wieckowski, Paolo Pinton, Valeria Poli
STAT3 mediates cytokine and growth factor receptor signalling, becoming transcriptionally active upon tyrosine 705 phosphorylation (Y-P). Constitutively Y-P STAT3 is observed in many tumors that become addicted to its activity, and STAT3 transcriptional activation is required for tumor transformation downstream of several oncogenes. We have recently demonstrated that constitutively active STAT3 drives a metabolic switch towards aerobic glycolysis through the transcriptional induction of Hif-1α and the down-regulation of mitochondrial activity, in both MEF cells expressing constitutively active STAT3 (Stat3C/C) and STAT3-addicted tumor cells...
2014: Cancers
Martine Blouin, Mathieu Lalumière, Dany H Gagnon, Félix Chénier, Rachid Aissaoui
Eighteen manual wheelchair users (MWUs) with spinal cord injury participated in a training session on a new manual wheelchair simulator with haptic biofeedback (HB). The training aimed to modify participants' mechanical effective force (MEF) along the push phase to achieve a target MEF pattern slightly more effective than their pre-training pattern. More HB was provided if the participants' achieved MEF pattern deviated from the target. Otherwise, less HB was provided. The deviation between the participants' achieved MEF and the target, as well as the mean achieved MEF, were computed before, during and after the training session...
January 2015: IEEE Transactions on Neural Systems and Rehabilitation Engineering
Khalid Amine Timani, Ying Liu, Attaya Suvannasankha, Johnny J He
Tip110 is a nuclear protein and has been shown to function in tumor antigenicity, regulation of gene transcription, pre-mRNA splicing, stem cell proliferation and differentiation, and embryonic development. To characterize the in vivo functions of Tip110, a transgene cassette expressing human Tip110 protein (hTip110) was used to generate hTip110 transgenic (Tg) mice. Unexpectedly, only Tip110 mRNA but not Tip110 protein was expressed in Tg MEF and tissues. Treatment of Tg MEF with proteasome inhibitors led to detection of hTip110 protein, which prompted us to investigate the regulatory mechanisms of Tip110 degradation in mouse cells...
September 2014: International Journal of Biochemistry & Cell Biology
Hiu Yeung Lau, Pondy M Ramanujulu, Dianyan Guo, Tianming Yang, Melissa Wirawan, Patrick J Casey, Mei-Lin Go, Mei Wang
Inhibitors of isoprenylcysteine carboxylmethyltransferase (Icmt) are promising anti-cancer agents, as modification by Icmt is an essential component of the protein prenylation pathway for a group of proteins that includes Ras GTPases. Cysmethynil, a prototypical indole-based inhibitor of Icmt, effectively inhibits tumor cell growth. However, the physical properties of cysmethynil, such as its low aqueous solubility, make it a poor candidate for clinical development. A novel amino-derivative of cysmethynil with superior physical properties and marked improvement in efficacy, termed compound 8...
September 2014: Cancer Biology & Therapy
Ming-Jen Sheu, Chien-Feng Li, Ching-Yih Lin, Sung-Wei Lee, Li-Ching Lin, Tzu-Ju Chen, Li-Jung Ma
Neoadjuvant concurrent chemoradiation therapy (CCRT) is an increasingly common therapeutic strategy for rectal cancer. Clinically, it remains a major challenge to predict therapeutic response and patient outcomes after CCRT. Annexin I (ANXA1), encoded by ANXA1, is a Ca(2+)/phospholipid-binding protein that mediates actin dynamics and cellular proliferation, as well as suggesting tumor aggressiveness and predicting therapeutic response in certain malignancies. However, expression of ANXA1 has never been reported in rectal cancer receiving CCRT...
August 2014: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
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