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J Kabutomori, O Beloto-Silva, R R Geyer, R Musa-Aziz
Xenopus laevis oocytes are a valuable tool for investigating the function of membrane proteins. However, regulations around the world, specifically in Brazil, render the import of Xenopus laevis frogs impractical, and, in some cases, impossible. Here, as an alternative, we evaluate the usefulness of the North American aquatic bullfrog Lithobates catesebeianus , which is commercially available in Brazil, for the heterologous expression of aquaporin (AQP) proteins. We have developed a method that combines a brief collagenase treatment and mechanical defolliculation for isolating individual oocytes from Lithobates ovaries...
March 12, 2018: Biology Open
Molly H B Amador, M Danielle McDonald
The serotonin transporter (SERT) functions in the uptake of the neurotransmitter serotonin (5-HT) from the extracellular milieu and is the molecular target of the selective serotonin reuptake inhibitors (SSRIs), a common group of antidepressants. The current study comprehensively assesses the sequence, tissue distribution, transport kinetics, and physiological function of a teleost SERT. The 2,022-bp toadfish SERT sequence encodes a protein of 673 amino acids, which shows 83% similarity to zebrafish SERT and groups with SERT of other teleosts in phylogenetic analysis...
February 27, 2018: Journal of Experimental Biology
Evan J Myers, Aniko Marshall, Mark D Parker
Mutations in the sodium bicarbonate cotransporter NBCe1 (SLC4A4) cause proximal renal tubular acidosis (pRTA). We recently described a novel pRTA mutation p.Gln913Arg (Q913R), inherited in compound heterozygous form with p.Arg510His (R510H). Q913R causes intracellular retention of NBCe1 and a 'gain of function' Cl - leak. To learn more about the importance of glutamine at position 913, we substituted a variety of alternative amino-acid residues (Cys, Glu, Lys, Leu, Ser) at position 913. Studying cRNA-injected Xenopus oocytes by voltage clamp, we find that most de novo mutants exhibit close-to-normal NBCe1 activity; only Q913K expresses a Cl - leak...
February 15, 2018: Scientific Reports
Sunil More, Xiaoyun Yang, Zhengyu Zhu, Gayan Bamunuarachchi, Yujie Guo, Chaoqun Huang, Keith Bailey, Jordan P Metcalf, Lin Liu
Wnt/β-catenin signaling is an essential pathway in cell cycle control. Dysregulation of the Wnt/β-catenin signaling pathway during viral infection has been reported. In this study, we examined the effect of modulating Wnt/β-catenin signaling during influenza virus infection. The activation of the Wnt/β-catenin pathway by Wnt3a increased influenza virus mRNA and virus production in in vitro in mouse lung epithelial E10 cells and mRNA expresson of influenza virus genes in vivo in the lungs of mice infected with influenza virus A/Puerto Rico/8/34...
2018: PloS One
Yi-Hsuan Wu, Ying-Hsuan Lee, Hung-Yu Shih, Shih-Hsiang Chen, Yi-Chuan Cheng, Daniel Tsun-Yee Chiu
Glucose-6-phosphate dehydrogenase (G6PD) is a housekeeping enzyme involved in the pentose phosphate shunt for producing nicotinamide adenine dinucleotide phosphate (NADPH). Severe G6PD deficiency leads to embryonic lethality, but the underlying mechanism is unclear. In the current study, the effects of G6PD on epithelial-mesenchymal transition (EMT), especially during embryonic development, were investigated. The knockdown of G6PD induced morphological changes, accompanied by the suppression of epithelial markers, E-cadherin and β-catenin, in A549 and MDCK cells...
January 9, 2018: Cell Death & Disease
Yukiko Nakamura, Yusuke Ishida, Makoto Kondo, Shoichi Shimada
The serotonin (5-hydroxytryptamine) type 3 (5-HT3) receptor is an important target in the control of emesis, and 5-HT3 receptor antagonists are effective against the early phase chemotherapy evoked vomiting. We recently reported that the anticancer drugs irinotecan and topotecan directly modulate the 5-HT-mediated 5-HT3 receptor current in vitro. In addition, the drug response depends on the 5-HT3 subunit composition. Here, we explored the effects of 35 anticancer drugs on the 5-HT3 receptor current. We microinjected Xenopus laevis oocytes with human 5-HT3A cRNA or a combination of human 5-HT3A and human 5-HT3B cRNA, and performed two-electrode voltage clamp recordings of 5-HT3A and 5-HT3AB receptor currents in the presence of each of the 35 drugs...
December 22, 2017: European Journal of Pharmacology
Wataru Shibata, Soichiro Sue, Sachiko Tsumura, Yasuaki Ishii, Takeshi Sato, Eri Kameta, Makoto Sugimori, Hiroaki Yamada, Hiroaki Kaneko, Tomohiko Sasaki, Tomohiro Ishii, Toshihide Tamura, Masaaki Kondo, Shin Maeda
BACKGROUND: Although Helicobacter-induced gastric inflammation is the major predisposing factor for gastric carcinogenesis, the precise mechanism by which chronic gastritis causes gastric cancer remains unclear. Intestinal and spasmolytic polypeptide-expressing metaplasia (SPEM) is considered as precancerous lesions, changes in epithelial tissue stem/progenitor cells after chronic inflammation has not been clarified yet. In this study, we utilized three-dimensional gastric epithelial cell culture systems that could form organoids, mimicking gastric epithelial layer, and characterized the changes in epithelial cells after chronic Helicobacter felis infection...
December 6, 2017: BMC Gastroenterology
Na Yu, Yang Liu, Xuan Wang, Jian Li, Haibo Bao, Zewen Liu
Nicotinic acetylcholine receptor (nAChR) subunits are encoded by a large multigene family and generate a large number of pentameric receptors with various properties. At present, nematode species, such as Caenorhabditis elegans, have the largest number of nAChR subunits. In this study, two nAChR subunits (Bxy-Unc-38 and Bxy-Unc-29) were cloned from Bursaphelenchus xylophilus, a fatal nematode pest on pine trees causing pine wilt disease. When Bxy-Unc-38 and Bxy-Unc-29 were co-expressed in Xenopus oocytes, constructed functional nAChRs showed agonist responses to acetylcholine and imidacloprid, a neonicotinoid insecticide...
November 2017: Pesticide Biochemistry and Physiology
Makhala M Khammy, Sukhan Kim, Bo H Bentzen, Soojung Lee, Inyeoung Choi, Christian Aalkjaer, Thomas A Jepps
BACKGROUND AND PURPOSE: Kv7.4 and Kv7.5 channels are regulators of vascular tone. 4-aminopyridine (4-AP) is considered a broad inhibitor of voltage-gated potassium (KV ) channels, with little inhibitory effect on Kv7 family members at millimolar concentrations. However, the effect of 4-AP on Kv7 channels has not been systematically studied. The aim of this study was to investigate the pharmacological activity of 4-AP on Kv7.4 and Kv7.5 channels and characterise the effect of 4-AP on rat resistance arteries...
November 20, 2017: British Journal of Pharmacology
Betty Hoac, Delia Susan-Resiga, Rachid Essalmani, Edwige Marcinkiweicz, Nabil G Seidah, Marc D McKee
Seven proprotein convertases cleave the basic amino acid consensus sequence K/R-Xn -K/R↓ (where n=0, 2, 4 or 6 variable amino acids) to activate precursor proteins. Despite similarities in substrate specificity, basic amino acid-specific proprotein convertases have a distinct tissue distribution allowing for enzymatic actions on tissue-resident substrates. Proprotein convertase 5/6 (PC5/6) has two splice variants - soluble PC5/6A and membrane-bound PC5/6B - and is expressed during mouse development in many tissues including bone and tooth, but little is known about the substrates for PC5/6 therein...
February 2018: Bone
Judith Oymans, Aartjan J W Te Velthuis
The influenza A virus genome consists of eight segments of single-stranded RNA. These segments are replicated and transcribed by a viral RNA-dependent RNA polymerase (RdRp) that is made up of the influenza virus proteins PB1, PB2 and PA. To copy the viral RNA (vRNA) genome segments and the complementary RNA (cRNA) segments, the replicative intermediate of viral replication, the RdRp must use two promoters and two different de novo initiation mechanisms. On the vRNA promoter, the RdRp initiates on the 3' terminus, while on the cRNA promoter the RdRp initiates internally and subsequently realigns the nascent vRNA product to ensure that the template is copied in full...
November 8, 2017: Journal of Virology
Simona Jarc, Miloš Miler, Stanka Šebela, Nina Zupančič
In the caves, the formation of cave minerals is a consequence of a variety of chemical reactions, some of them also due to human activity. There are many caves in Slovenia, but sulphate minerals are not very often reported and analysed. In this study, the presence of sulphate minerals is detected by SEM/EDS analysis of speleothems from Črna Jama, a cave near Kočevje (southern Slovenia). The cave is characterised by its dark, almost black colour on cave walls, floor and speleothems. Anthropogenic influence in the cave is still visible, including the remains of a fireplace, some inscriptions on the walls and wooden containers...
December 2017: Environmental Science and Pollution Research International
Laurie J Goldsmith, Ashra Kolhatkar, Dominic Popowich, Anne M Holbrook, Steven G Morgan, Michael R Law
Many patients report skipping doses, splitting pills, or not filling prescriptions due to out-of-pocket costs-a phenomenon known as cost-related non-adherence (CRNA). This study investigated CRNA from the patient's perspective, and, to our knowledge, is the first study to undertake a qualitative investigation of CRNA specifically. We report the results from 35 semi-structured interviews conducted in 2014-15 with adults in four Canadian cities across two provinces. We used framework analysis to develop a CRNA typology to characterize major factors in patients' CRNA decisions...
October 9, 2017: Social Science & Medicine
Kazuhiro Ishibashi, Eiko Matsumoto-Yokoyama, Masayuki Ishikawa
Tomato spotted wilt virus (TSWV) is a negative-strand RNA virus of the order Bunyavirales, family Tospoviridae, genus Orthotospovirus. TSWV infects a broad range of plant species, causing serious economic losses. Despite its agronomic importance, molecular biological understanding of TSWV has been limited, partly due to the lack of a reverse genetics system, which would enable genetic manipulation of the virus. Here, we report that RNA synthesis by TSWV RNA polymerase occurs in the yeast Saccharomyces cerevisiae using a segment of the TSWV genome, S RNA expressed from cloned cDNA, as a template...
October 4, 2017: Scientific Reports
J H Kim, K J Gellatly, B Lueke, M Kohler, R Nauen, E Murenzi, K S Yoon, J M Clark
We previously observed that ivermectin-induced detoxification genes, including ATP binding cassette transporter C4 (PhABCC4) and cytochrome P450 6CJ1 (CYP6CJ1) were identified from body lice following a brief exposure to a sublethal dose of ivermectin using a non-invasive induction assay. In this current study, the functional properties of PhABCC4 and CYP6CJ1 were investigated after expression in either X. laevis oocytes or using a baculovirus expression system, respectively. Efflux of [(3) H]-9-(2-phosphonomethoxyethyl) adenine ([(3) H]-PMEA), a known ABCC4 substrate in humans, was detected from PhABCC4 cRNA-injected oocytes by liquid scintillation spectrophotometric analysis and PhABCC4 expression in oocytes was confirmed using ABC transporter inhibitors...
September 27, 2017: Insect Molecular Biology
Mary Scott-Herring, Sarabdeep Singh
Little evidence exists to support the effects of a preceptor-mentor program for orienting newly hired certified registered nurse anesthetists (CRNAs). The literature suggests that participation in a preceptorship-mentorship program will increase the satisfaction, confidence, and comfort of both preceptors and newly hired orientees. The purpose of this project was to determine the effects of these outcomes. This program was developed based on best evidence and implemented as a quality improvement project. Three sessions were held to educate preceptors (N = 12) in the following areas: communication skills, providing constructive feedback, and assessing learner's styles and needs...
October 1, 2017: Journal of Continuing Education in Nursing
Beatriz Quiñones, Bertram G Lee, Todd J Martinsky, Jaszemyn C Yambao, Paul K Haje, Mark Schena
Human noroviruses (NoV) are the leading cause of human gastroenteritis in populations of all ages and are linked to most of the foodborne outbreaks worldwide. Hepatitis A virus (HAV) is another important foodborne enteric virus and is considered the most common agent causing acute liver disease worldwide. In the present study, a focused, low-density DNA microarray was developed and validated for the simultaneous identification of foodborne-associated genotypes of NoV and HAV. By employing a novel algorithm, capture probes were designed to target variable genomic regions commonly used for typing these foodborne viruses...
September 20, 2017: Sensors
Lesley J J Soril, Ted Adams, Madeleine Phipps-Taylor, Ulrika Winblad, Fiona Clement
OBJECTIVE: To compare cost-related non-adherence (CRNA), serious problems paying medical bills and average annual out-of-pocket cost over time in five countries. METHODS: Repeated cross-sectional analysis of the Commonwealth Fund International Health Policy survey from 2004 to 2014. Responses were compared between Canada, the UK, Australia, New Zealand and the US. RESULTS: Compared to the UK, respondents in Canada, Australia and New Zealand were two to three times and respondents in the US were eight times more likely to experience CRNA; these odds remained stable over time...
August 2017: Healthcare Policy, Politiques de Santé
Wanyin Tao, Tianyu Gan, Mingzhe Guo, Yongfen Xu, Jin Zhong
Ebola virus (EBOV) causes severe hemorrhagic fever in humans and other primates with a high case fatality rate. No approved drug or vaccine of EBOV is available, which necessitates better understanding of the virus life cycle. Studies on EBOV have been hampered because experimentations involving live virus are restricted to biosafety level 4 (BSL-4) laboratories. EBOV minigenome system has provided researchers with the opportunity to study EBOV under BSL-2 conditions. Here, we developed a novel EBOV minigenome replicon which, to our knowledge, is the first EBOV cell culture system that can stably replicate and transcribe EBOV minigenome...
September 6, 2017: Journal of Virology
Donald Boyd, Lusine Poghosyan
BACKGROUND AND PURPOSE: No tool exists measuring certified registered nurse anesthetist (CRNA) organizational climate. The study's purpose is to adapt a validated tool to measure CRNA organizational climate. METHODS: Content validity of the Certified Registered Nurse Anesthetist Organizational Climate Questionnaire (CRNA-OCQ) was established. Pilot testing was conducted to determine internal reliability consistency of the subscales. RESULTS: Experts rated the tool as content valid...
August 1, 2017: Journal of Nursing Measurement
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