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Hjalte C R Sass, Rehannah Borup, Mikkel Alanin, Finn Cilius Nielsen, Per Cayé-Thomasen
The objective of this study was to determine global gene expression in relation to Vestibular schwannomas (VS) growth rate and to identify signal transduction pathways and functional molecular networks associated with growth. Repeated magnetic resonance imaging (MRI) prior to surgery determined tumor growth rate. Following tissue sampling during surgery, mRNA was extracted from 16 sporadic VS. Double stranded cDNA was synthesized from the mRNA and used as template for in vitro transcription reaction to synthesize biotin-labeled antisense cRNA, which was hybridized to Affymetrix HG-U133A arrays and analyzed by dChip software...
October 17, 2016: Journal of Neuro-oncology
Sylvia J Bedford-Guaus, François Chauvigné, Eva Mejía-Ramírez, Mercè Martí, Antoni Ventura-Rubio, Ángel Raya, Joan Cerdà, Anna Veiga
While vitrification has become the method of choice for preservation of human oocytes and embryos, cryopreservation of complex tissues and of large yolk-containing cells, remains largely unsuccessful. One critical step in such instances is appropriate permeation while avoiding potentially toxic concentrations of cryoprotectants. Permeation of water and small non-charged solutes, such as those used as cryoprotectants, occurs largely through membrane channel proteins termed aquaporins (AQPs). Substitution of a Thr by an Ala residue in the pore-forming motif of the zebrafish (Dario rerio) Aqp3b paralog resulted in a mutant (DrAqp3b-T85A) that when expressed in Xenopus or porcine oocytes increased their permeability to ethylene glycol at pH 7...
October 5, 2016: Zygote: the Biology of Gametes and Early Embryos
Nicole C Robb, Aartjan J W Te Velthuis, Ralph Wieneke, Robert Tampé, Thorben Cordes, Ervin Fodor, Achillefs N Kapanidis
Influenza viruses have a segmented viral RNA (vRNA) genome, which is replicated by the viral RNA-dependent RNA polymerase (RNAP). Replication initiates on the vRNA 3' terminus, producing a complementary RNA (cRNA) intermediate, which serves as a template for the synthesis of new vRNA. RNAP structures show the 3' terminus of the vRNA template in a pre-initiation state, bound on the surface of the RNAP rather than in the active site; no information is available on 3' cRNA binding. Here, we have used single-molecule Förster resonance energy transfer (smFRET) to probe the viral RNA conformations that occur during RNAP binding and initial replication...
September 30, 2016: Nucleic Acids Research
James Kirui, Arindam Mondal, Andrew Mehle
: The influenza A virus polymerase plays an essential role in the virus lifecycle, directing synthesis of viral mRNAs and genomes. It is a trimeric complex composed of subunits PA, PB1, and PB2 and associates with viral RNAs and nucleoprotein (NP) to form higher order ribonucleoprotein (RNP) complexes. The polymerase is regulated temporally over the course of infection to ensure coordinated expression of viral genes as well as replication of the viral genome. Various host factors and processes have been implicated in regulation of the IAV polymerase function, including post-translational modifications, however the mechanisms are not fully understood...
September 28, 2016: Journal of Virology
Katy E French, Alexis B Guzman, Augustin C Rubio, John C Frenzel, Thomas W Feeley
BACKGROUND: With the movement towards bundled payments, stakeholders should know the true cost of the care they deliver. Time-driven activity-based costing (TDABC) can be used to estimate costs for each episode of care. In this analysis, TDABC is used to both estimate the costs of anesthesia care and identify the primary drivers of those costs of 11 common oncologic outpatient surgical procedures. METHODS: Personnel cost were calculated by determining the hourly cost of each provider and the associated process time of the 11 surgical procedures...
September 2016: Healthcare
R Al-Kattan, M Retzepi, E Calciolari, N Donos
OBJECTIVES: To investigate the gene expression and molecular pathways implicated in the regulation of the osseous healing process following guided bone regeneration (GBR). MATERIAL AND METHODS: Six 6-month-old Wistar male rats were used. Standardized 5-mm critical size defects were created in the parietal bones of each animal and treated with an extracranial and intracranial ePTFE membrane, according to the GBR principle. Three animals were randomly sacrificed after 7 and 15 days of healing...
September 11, 2016: Clinical Oral Implants Research
Erika Moreno, Consuelo Plata, Alejandro Rodríguez-Gama, Eduardo R Argaiz, Norma Vázquez, Karla Leyva-Ríos, León Islas, Christopher Cutler, Diana Pacheco-Alvarez, Adriana Mercado, Raquel Cariño, María Castañeda-Bueno, Gerardo Gamba
The thiazide-sensitive Na-Cl cotransporter NCC is the major pathway for salt reabsorption in the mammalian distal convoluted tubule. NCC plays a key role in the regulation of blood pressure. Its inhibition with thiazides constitutes primary baseline therapy for arterial hypertension. However, the thiazide-binding site in NCC is unknown. Mammals have only one gene encoding for NCC. The eel, however, contains a duplicate gene. NCCα is an ortholog of mammalian NCC and is expressed in the kidney. NCCβ is present in the apical membrane of the rectum...
September 1, 2016: Journal of Biological Chemistry
Myriam Fezai, Musaab Ahmed, Zohreh Hosseinzadeh, Florian Lang
BACKGROUND/AIMS: The pleotropic functions of the large conductance Ca2+-activated K+ channels (maxi K+ channel or BK channels) include regulation of neuronal excitation and cell volume. Kinases participating in those functions include the glycogen synthase kinase GSK3 ß which is under negative control of protein kinase B (PKB/Akt). GSK3ß is inhibited by the antidepressant Lithium. The present study thus explored whether GSK3ß modifies the activity of BK channels. METHODS: cRNA encoding the Ca2+ insensitive BK channel mutant BKM513I+Δ899-903 was injected into Xenopus laevis oocytes without or with additional injection of cRNA encoding wild-type GSK3ß, inactive K85RGSK3ß, or wild-type GSK3ß with wild-type PKB...
2016: Cellular Physiology and Biochemistry
Adriana López-Barradas, Tania González-Cid, Norma Vazquez, Marisol Gavi-Maza, Adriana Reyes-Camacho, Laura Alejandra Velazquez-Villegas, Victoria Ramirez, Kambiz Zandi-Nejad, David B Mount, Nimbe Torres, Armando R Tovar, Michael F Romero, Gerardo Gamba, Consuelo Plata
SMCTs move several important fuel molecules that are involved in lipid, carbohydrate, and amino acid metabolism, but their regulation has been poorly studied. Insulin controls the translocation of several solutes that are involved in energetic cellular metabolism, including glucose. We studied the effect of insulin on the function of human SMCT1 expressed in Xenopus oocytes. The addition of insulin reduced α-keto-isocaproate (KIC)-dependent (22)Na(+) uptake by 29%. Consistent with this result, the co-injection of SMCT1 with SGK1 cRNA decreased the KIC-dependent (22)Na(+) uptake by 34%...
August 3, 2016: American Journal of Physiology. Cell Physiology
Sun Li, Xiao-Lin Liu, Xie-Lai Zhou, Su-Jun Jiang, Hong Yuan
The toxic metal lead is a widespread environmental polutant that can adversely affect human health. However, the underlying mechanisms of lead-induced toxicity are still largely unknown. The mechanism of lead toxicity was presumed to involve cross reaction between Pb(2+) and Ca(2+) with calmodulin dependent systems. The aim of the present study was thus to identify differential expression of calmodulin-related genes in the spleen of lead-exposed mice. We performed microarray analysis to identify differentially expressed genes...
December 2015: Interdisciplinary Toxicology
Rajendra Boggavarapu, Stephan Hirschi, Daniel Harder, Marcel Meury, Zöhre Ucurum, Marc J Bergeron, Dimitrios Fotiadis
This protocol describes the isolation of recombinant human and mammalian membrane proteins expressed in Xenopus laevis frog oocytes for structural studies. The cDNA-derived cRNA of the desired genes is injected into several hundreds of oocytes, which are incubated for several days to allow protein expression. Recombinant proteins are then purified via affinity chromatography. The novelty of this method comes from the design of a plasmid that produces multi-tagged proteins and, most importantly, the development of a protocol for efficiently discarding lipids, phospholipids, and lipoproteins from the oocyte egg yolk, which represent the major contaminants in protein purifications...
2016: Methods in Molecular Biology
Pu Gao, Hui Yang, Kanagalaghatta R Rajashankar, Zhiwei Huang, Dinshaw J Patel
CRISPR-Cas9 and CRISPR-Cpf1 systems have been successfully harnessed for genome editing. In the CRISPR-Cas9 system, the preordered A-form RNA seed sequence and preformed protein PAM-interacting cleft are essential for Cas9 to form a DNA recognition-competent structure. Whether the CRISPR-Cpf1 system employs a similar mechanism for target DNA recognition remains unclear. Here, we have determined the crystal structure of Acidaminococcus sp. Cpf1 (AsCpf1) in complex with crRNA and target DNA. Structural comparison between the AsCpf1-crRNA-DNA ternary complex and the recently reported Lachnospiraceae bacterium Cpf1 (LbCpf1)-crRNA binary complex identifies a unique mechanism employed by Cpf1 for target recognition...
August 2016: Cell Research
Ziyu Liu, Fang Wang, Lijuan Yuan, Xiaoxiao Zhang, Qikang Ying, Lan Yu, Liang Zhang, Linfeng Cheng, Fanglin Zhang, Jianguo Lu, Xing'an Wu
Hemorrhagic fever with renal syndrome (HFRS) is a severe, viral zoonotic disease which occurs worldwide, particularly in Asia and Europe. In China, the Hantaan virus (HTNV) and the Seoul virus (SEOV) are known to be the most prevalent causative agents of HFRS. Since no protective vaccines or effective treatments are available for human use, accurate and reliable diagnostic methods are essential for disease surveillance. In the present study, the viral loads in cell culture supernatant, infected mice blood and clinical serum samples were quantified using the SYBR‑Green I-based reverse transcription-quantitiative polymerase chain reaction (RT-qPCR) assay, which targeted the S gene sequence of the HTNV and SEOV genomes...
September 2016: International Journal of Molecular Medicine
Emmanuel Olushola Shobowale, Benjamin Adegunle, Ken Onyedibe
BACKGROUND: Hand hygiene has been described as the cornerstone and starting point in all infection control programs, with the hands of healthcare staff being the drivers and promoters of infection in critically ill patients. The objectives of this study were to access healthcare workers compliance with the World Health Organization (WHO) prescribed five moments of hand hygiene as it relates to patient care and to determine the various strata of healthcare workers who are in default of such prescribed practices...
May 2016: Nigerian Medical Journal: Journal of the Nigeria Medical Association
Ruth D E Fritsch-Stork, Sandra C Silva-Cardoso, Marjan J A Groot Koerkamp, Jasper C A Broen, Floris F P Lafeber, Johannes W J Bijlsma Bijlsma
OBJECTIVES: Glucocorticoids (GC) remain a cornerstone of rheumatoid arthritis (RA) therapy, although a third of patients do not respond adequately. In order to find potential predictors for clinical response, the gene expression profile of CD4+T-cells as important players in the pathogenesis of RA was analysed before pulse therapy with 1000 mg methylprednisolone. METHODS: Patients were treated with 3x1000 mg methylprednisolone in 5 days; hereafter response was determined by the European League Against Rheumatism (EULAR) response criteria...
July 2016: Clinical and Experimental Rheumatology
Nisa Wongsamitkul, Roland Baur, Erwin Sigel
GABAA receptors are pentameric ligand-gated channels mediating inhibitory neurotransmission in the CNS. α4βxδ GABAA receptors are extrasynaptic receptors important for tonic inhibition. The functional properties and subunit arrangement of these receptors are controversial. We predefined subunit arrangement by using subunit concatenation. α4, β2, and δ subunits were concatenated to dimeric, trimeric, and, in some cases, pentameric subunits. We constructed in total nine different receptor pentamers in at least two different ways and expressed them in Xenopus oocytes...
August 26, 2016: Journal of Biological Chemistry
Lauren R Snoots, Brenda A Wands
Certified Registered Nurse Anesthetists (CRNAs) provide high-quality patient care to ensure patient safety. Strict vigilance and prompt response is required of the CRNA to make critical decisions. Distractions during anesthesia delivery may threaten patient safety. Personal electronic devices (PEDs) have become an integral tool used by 90% of adults. Adaptation of PEDs has permitted their integration into current anesthesia practice. Although technologic advancements have improved accessibility to resources and communication, they also serve as a source of distraction...
April 2016: AANA Journal
Joanna Laskowska, Joanna Lewandowska-Bieniek, Joanna Szczepanek, Jan Styczyński, Andrzej Tretyn
BACKGROUND: A major problem in the treatment of leukemia is the development of drug resistance to chemotherapeutic agents. METHODS: To determine the ex vivo drug resistance profile to anthracyclines, an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT) cytotoxicity assay was performed on mononuclear cells obtained from 155 patients with acute lymphoblastic leukemia (ALL) or acute myeloblastic leukemia (AML). Gene expression profiles (for 51 patients with ALL and 16 with AML) were prepared on the basis of cRNA hybridization to oligonucleotide arrays of the human genome (Affymetrix)...
August 2016: Journal of Gene Medicine
Melanie Abongwa, Samuel K Buxton, Elise Courtot, Claude L Charvet, Cédric Neveu, Ciaran J McCoy, Saurabh Verma, Alan P Robertson, Richard J Martin
BACKGROUND AND PURPOSE: Control of nematode parasite infections relies largely on anthelmintic drugs, several of which act on nicotinic ACh receptors (nAChRs), and there are concerns about the development of resistance. There is an urgent need for development of new compounds to overcome resistance and novel anthelmintic drug targets. We describe the functional expression and pharmacological characterization of a homomeric nAChR, ACR-16, from a nematode parasite. EXPERIMENTAL APPROACH: Using RT-PCR, molecular cloning and two-electrode voltage clamp electrophysiology, we localized acr-16 mRNA in Ascaris suum (Asu) and then cloned and expressed acr-16 cRNA in Xenopus oocytes...
August 2016: British Journal of Pharmacology
Lisa L Hall, Jeanne B Lawrence
Beginning with the precedent of XIST RNA as a 'chromosomal RNA' (cRNA), there is growing interest in the possibility that a diversity of non-coding RNAs may function in chromatin. We review findings which lead us to suggest that RNA is essentially a widespread component of interphase chromosomes. Further, RNA likely contributes to architecture and regulation, with repeat-rich 'junk' RNA in euchromatin (ecRNA) promoting a more open chromatin state. Thousands of low-abundance nuclear RNAs have been reported, however it remains a challenge to determine which of these may function in chromatin...
April 2016: Current Opinion in Genetics & Development
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