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functional screening

Huiting Zhao, Yali Du, Pengfei Gao, Shujie Wang, Jianfang Pan, Yusuo Jiang
Chemosensory genes play a central role in sensing chemical signals and guiding insect behavior. The Chinese honeybee, Apis cerana cerana, is one of the most important insect species in China in terms of resource production, and providing high-quality products for human consumption, and also serves as an important pollinator. Communication and foraging behavior of worker bees is likely linked to a complex chemosensory system. Here, we used transcriptome sequencing on adult A. c. cerana workers of different ages to identify the major chemosensory gene families and the differentially expressed genes(DEGs), and to investigate their expression profiles...
2016: PloS One
Cheng-Wei Wu, Andrew Deonarine, Aaron Przybysz, Kevin Strange, Keith P Choe
SKN-1/Nrf are the primary antioxidant/detoxification response transcription factors in animals and they promote health and longevity in many contexts. SKN-1/Nrf are activated by a remarkably broad-range of natural and synthetic compounds and physiological conditions. Defining the signaling mechanisms that regulate SKN-1/Nrf activation provides insights into how cells coordinate responses to stress. Nrf2 in mammals is regulated in part by the redox sensor repressor protein named Keap1. In C. elegans, the p38 MAPK cascade in the intestine activates SKN-1 during oxidative stress by promoting its nuclear accumulation...
October 2016: PLoS Genetics
Yuchen Gao, Xin Xiong, Spencer Wong, Emeric J Charles, Wendell A Lim, Lei S Qi
The ability to dynamically manipulate the transcriptome is important for studying how gene networks direct cellular functions and how network perturbations cause disease. Nuclease-dead CRISPR-dCas9 transcriptional regulators, while offering an approach for controlling individual gene expression, remain incapable of dynamically coordinating complex transcriptional events. Here, we describe a flexible dCas9-based platform for chemical-inducible complex gene regulation. From a screen of chemical- and light-inducible dimerization systems, we identified two potent chemical inducers that mediate efficient gene activation and repression in mammalian cells...
October 24, 2016: Nature Methods
Chip P Rowan, Michael C Riddell, Norman Gledhill, Veronica K Jamnik
PURPOSE: Prediabetes is linked to several modifiable risk factors, in particular, physical activity participation. The optimal prescription for physical activity remains uncertain. This pilot study aimed to investigate the effectiveness of continuous moderate intensity (CON) versus high intensity interval (HIIT) aerobic training in persons with prediabetes. Outcome measures included hemoglobin A1c (A1C), body composition, musculoskeletal and aerobic fitness. METHODS: Participants (n= 35) were recruited and screened using a questionnaire plus capillary blood point-of-care A1C analysis...
October 21, 2016: Medicine and Science in Sports and Exercise
Dai Horiuchi, Roman Camarda, Alicia Y Zhou, Christina Yau, Olga Momcilovic, Sanjeev Balakrishnan, Alexandra N Corella, Henok Eyob, Kai Kessenbrock, Devon A Lawson, Lindsey A Marsh, Brittany N Anderton, Julia Rohrberg, Ratika Kunder, Alexey V Bazarov, Paul Yaswen, Michael T McManus, Hope S Rugo, Zena Werb, Andrei Goga
Triple-negative breast cancer (TNBC), in which cells lack expression of the estrogen receptor (ER), the progesterone receptor (PR) and the ERBB2 (also known as HER2) receptor, is the breast cancer subtype with the poorest outcome. No targeted therapy is available against this subtype of cancer owing to a lack of validated molecular targets. We previously reported that signaling involving MYC-an essential, pleiotropic transcription factor that regulates the expression of hundreds of genes-is disproportionally higher in triple-negative (TN) tumors than in receptor-positive (RP) tumors...
October 24, 2016: Nature Medicine
M Vouri, D R Croucher, S P Kennedy, Q An, G J Pilkington, S Hafizi
Acquired resistance to conventional and targeted therapies is becoming a major hindrance in cancer management. It is increasingly clear that cancer cells are able to evolve and rewire canonical signalling pathways to their advantage, thus evading cell death and promoting cell invasion. The Axl receptor tyrosine kinase (RTK) has been shown to modulate acquired resistance to EGFR-targeted therapies in both breast and lung cancers. Glioblastoma multiforme (GBM) is a highly infiltrative and invasive form of brain tumour with little response to therapy...
October 24, 2016: Oncogenesis
Ken-Ichiro Kamei, Yoshie Koyama, Yumie Tokunaga, Yasumasa Mashimo, Momoko Yoshioka, Christopher Fockenberg, Rowland Mosbergen, Othmar Korn, Christine Wells, Yong Chen
Human pluripotent stem cells hold great promise for applications in drug discovery and regenerative medicine. Microfluidic technology is a promising approach for creating artificial microenvironments; however, although a proper 3D microenvironment is required to achieve robust control of cellular phenotypes, most current microfluidic devices provide only 2D cell culture and do not allow tuning of physical and chemical environmental cues simultaneously. Here, the authors report a 3D cellular microenvironment plate (3D-CEP), which consists of a microfluidic device filled with thermoresponsive poly(N-isopropylacrylamide)-β-poly(ethylene glycol) hydrogel (HG), which enables systematic tuning of both chemical and physical environmental cues as well as in situ cell monitoring...
October 24, 2016: Advanced Healthcare Materials
N Garg, D Bakhshinyan, C Venugopal, S Mahendram, D A Rosa, T Vijayakumar, B Manoranjan, R Hallett, N McFarlane, K H Delaney, J M Kwiecien, C C Arpin, P-S Lai, R F Gómez-Biagi, A M Ali, E D de Araujo, O A Ajani, J A Hassell, P T Gunning, S K Singh
Medulloblastoma (MB), the most common malignant paediatric brain tumor, is currently treated using a combination of surgery, craniospinal radiotherapy and chemotherapy. Owing to MB stem cells (MBSCs), a subset of MB patients remains untreatable despite standard therapy. CD133 is used to identify MBSCs although its functional role in tumorigenesis has yet to be determined. In this work, we showed enrichment of CD133 in Group 3 MB is associated with increased rate of metastasis and poor clinical outcome. The signal transducers and activators of transcription-3 (STAT3) pathway are selectively activated in CD133(+) MBSCs and promote tumorigenesis through regulation of c-MYC, a key genetic driver of Group 3 MB...
October 24, 2016: Oncogene
Kyung Suk Lee, Lucy E Lee, Erel Levine
Small animals such as the roundworm C. elegans are excellent models for studying bacterial infection and host response, as well as for genetic and chemical screens. A key methodology is the killing assay, in which the number of surviving animals is tracked as a function of the time post infection. This is a labor-intensive procedure, prone to human error and subjective choices, and often involves undesired perturbation to the animals and their environment. In addition, the survival of animals is just one aspect of a multi-dimensional complex biological process...
October 24, 2016: Scientific Reports
Liis Andresen, Vallo Varik, Yuzuru Tozawa, Steffi Jimmy, Stina Lindberg, Tanel Tenson, Vasili Hauryliuk
The stringent response is a central adaptation mechanism that allows bacteria to adjust their growth and metabolism according to environmental conditions. The functionality of the stringent response is crucial for bacterial virulence, survival during host invasion as well as antibiotic resistance and tolerance. Therefore, specific inhibitors of the stringent response hold great promise as molecular tools for disarming and pacifying bacterial pathogens. By taking advantage of the valine amino acid auxotrophy of the Bacillus subtilis stringent response-deficient strain, we have set up a High Throughput Screening assay for the identification of stringent response inhibitors...
October 24, 2016: Scientific Reports
Fang Chen, Weifeng Zhang, Junli Zhao, Peiyan Yang, Rui Ma, Haibin Xia
Objective To prepare Rev-erbβ knockout HEK293 cells using clustered regularly interspaced short palindromic repeats/Cas 9 nuclease (CRISPR/Cas9) gene editing technology. Methods The knock-in or knockout of Rev-erbβ gene could be realized by single-guide RNA (sgRNA)-mediated Cas9 cutting of target DNA, and followed by DNA homologous recombination or non-homologous end joining-mediated DNA repair. Firstly, four sgRNAs were designed for Rev-erbβ gene. The sgRNA1 and sgRNA2 with the higher activity were respectively used to construct pCMV-hCas9-U6-Rev-erbβ sgRNA1 and pCMV-hCas9-U6-Rev-erbβ sgRNA2...
November 2016: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
Patricia Yuste-Checa, Sandra Brasil, Alejandra Gámez, Jarl Underhaug, Lourdes R Desviat, Magdalena Ugarte, Celia Pérez-Cerdá, Aurora Martinez, Belén Pérez
The congenital disorder of glycosylation (CDG) due to phosphomannomutase 2 deficiency (PMM2-CDG), the most common N-glycosylation disorder, is a multisystem disease for which no effective treatment is available. The recent functional characterization of disease-causing mutations described in patients with PMM2-CDG led to the idea of a therapeutic strategy involving pharmacological chaperones (PC) to rescue PMM2 loss-of-function mutations. The present work describes the high-throughput screening, by differential scanning fluorimetry, of 10,000 low molecular weight compounds from a commercial library, to search for possible PCs for the enzyme PMM2...
October 24, 2016: Human Mutation
Ivan V Smirnov, Andrey V Golovin, Spyros D Chatziefthimiou, Anastasiya V Stepanova, Yingjie Peng, Olga I Zolotareva, Alexey A Belogurov, Inna N Kurkova, Natalie A Ponomarenko, Matthias Wilmanns, G Michael Blackburn, Alexander G Gabibov, Richard A Lerner
In vitro selection of antibodies from large repertoires of immunoglobulin (Ig) combining sites using combinatorial libraries is a powerful tool, with great potential for generating in vivo scavengers for toxins. However, addition of a maturation function is necessary to enable these selected antibodies to more closely mimic the full mammalian immune response. We approached this goal using quantum mechanics/molecular mechanics (QM/MM) calculations to achieve maturation in silico. We preselected A17, an Ig template, from a naïve library for its ability to disarm a toxic pesticide related to organophosphorus nerve agents...
October 2016: Science Advances
Mahnaz Abbasi, Mohammadali Zohal, Banafsheh Atapour, Zohreh Yazdi
Introduction. Chronic obstructive pulmonary disease (COPD) is a major cause of morbidity and mortality worldwide. Proper diagnosis of osteoporosis as a systemic adverse effect of COPD is of significant importance. The present study aimed at evaluating the prevalence of osteoporosis and its risk factors in men suffering from COPD in Qazvin (2014). Methods. This descriptive-analytical study was conducted on 90 patients with COPD using random sampling. Anthropometric data and results from physical examination were collected...
2016: International Journal of Chronic Diseases
François Vromman, Stéphanie Perrinet, Lena Gehre, Agathe Subtil
Chlamydiae are Gram negative bacteria that develop exclusively inside eukaryotic host cells, within a membrane-bounded compartment. Members of the family Chlamydiaceae, such as Chlamydia trachomatis, are pathogenic species infecting vertebrates. They have a very reduced genome and exploit the capacities of their host for their own development, mainly through the secretion of proteins tailored to interfere with eukaryotic processes, called effector proteins. All Chlamydiaceae possess genes coding for four to five effectors that share a domain of unknown function (DUF582)...
2016: Frontiers in Cellular and Infection Microbiology
Joan A Vaccaro, Fatma G Huffman
Background. Approximately 17% of children aged 6-11 years were classified as obese in the United States. Obesity adversely affects physical functioning and leads to reduced quality of life. Heart function for overweight and obese children has not been reported. Methods. Data for this study were from NHANES National Youth Fitness Survey (NNYFS) conducted in conjunction with the National Health and Nutrition Examination Survey (NHANES) in 2012. This study used data from children aged 6-12 (N = 732) that had the cardiorespiratory endurance measure, body mass index for age and sex, and dietary data (N = 682)...
2016: Journal of Obesity
Kiyotaka Yanagihara, Yoshiharu Kinugasa, Kinya Shirota, Yoshiaki Inoue, Hiroshige Ishii, Fumiyo Tsunoda, Masaaki Iwata, Shinobu Sugihara, Shinya Takeda, Masayuki Hirai, Asao Mimura, Masahiko Kato, Kazuhiro Yamamoto
AIMS: Inadequate self-care management is a leading cause of re-hospitalization in patients with heart failure (HF). Psychological factors such as some ego functions interfere with self-care behaviour modification, leading to poor outcomes in patients with several chronic diseases. However, characteristics of ego states in patients with repeated hospitalization for HF remain undefined. METHODS AND RESULTS: The present study enrolled 40 HF outpatients with previous history of HF hospitalization and receiving self-care management...
March 2016: ESC Heart Failure
Beatriz Pérez Montoro, Nabil Benomar, Leyre Lavilla Lerma, Sonia Castillo Gutiérrez, Antonio Gálvez, Hikmate Abriouel
A collection of 31 Lactobacillus pentosus strains isolated from naturally fermented Aloreña green table olives were screened in depth in the present study for their probiotic potential. Several strains could be considered promising probiotic candidates since they showed good growth capacity and survival under simulated gastro-intestinal conditions (acidic pH of 1.5, up to 4% of bile salts and 5 mM of nitrate), good ability to auto-aggregate which may facilitate their adhesion to host cells as multiple aggregates and the subsequent displacement of pathogens...
2016: Frontiers in Microbiology
Elisete P Rodrigues, Cleiton de Paula Soares, Patrícia G Galvão, Eddie L Imada, Jean L Simões-Araújo, Luc F M Rouws, André L M de Oliveira, Márcia S Vidal, José I Baldani
Gluconacetobacter diazotrophicus is a beneficial nitrogen-fixing endophyte found in association with sugarcane plants and other important crops. Beneficial effects of G. diazotrophicus on sugarcane growth and productivity have been attributed to biological nitrogen fixation process and production of phytohormones especially indole-3-acetic acid (IAA); however, information about the biosynthesis and function of IAA in G. diazotrophicus is still scarce. Therefore, the aim of this work was to identify genes and pathways involved in IAA biosynthesis in this bacterium...
2016: Frontiers in Microbiology
Katie M de Almondes, Mônica V Costa, Leandro F Malloy-Diniz, Breno S Diniz
Aim: In this manuscript, we report data on the association between executive functions screened by Frontal Assessment Battery, Five Digit Test and Digit Span with self-reported depressive symptoms and sleep complaints in non-demented older adults. Methods: A total sample of 95 non-demented older adults performed Geriatric Depression Scale short version, Frontal Assessment Battery, Five Digit Test, Digit Span, and clinical interview. We split participants in groups stratified by age according to: young-old (60-69 years of age), old-old (70-79 years), and oldest-old (>80 years) and compared these three groups on the sociodemographic characteristics and executive functions performance...
2016: Frontiers in Psychology
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