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https://www.readbyqxmd.com/read/28109425/beyond-the-natural-proteome-nondegenerate-saturation-mutagenesis-methodologies-and-advantages
#1
M M Ferreira Amaral, L Frigotto, A V Hine
Beyond the natural proteome, high-throughput mutagenesis offers the protein engineer an opportunity to "tweak" the wild-type activity of a protein to create a recombinant protein with required attributes. Of the various approaches available, saturation mutagenesis is one of the core techniques employed by protein engineers, and in recent times, nondegenerate saturation mutagenesis is emerging as the approach of choice. This review compares the current methodologies available for conducting nondegenerate saturation mutagenesis with traditional, degenerate saturation and briefly outlines the options available for screening the resulting libraries, to discover a novel protein with the required activity and/or specificity...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28108582/identification-of-a-novel-protein-kinase-that-affects-the-chronological-lifespan-in-fission-yeast
#2
Tatsuhiro Kurauchi, Aya Hashizume, Yuki Imai, Kanako Hayashi, Satoshi Tsubouchi, Kunio Ihara, Hokuto Ohtsuka, Hirofumi Aiba
Chronological lifespan is defined by how long a cell can survive in a non-dividing state. In yeast, it is measured by viability after entry into the stationary phase. To understand the regulatory mechanisms of chronological lifespan in Schizosaccharomyces pombe, it is necessary to identify and characterize novel factors involved in the regulation of chronological lifespan. To this end, we have screened for a long-lived mutant and identified that novel gene nnk1(+) that encodes an essential protein kinase is the determinant of chronological lifespan...
January 2017: FEMS Microbiology Letters
https://www.readbyqxmd.com/read/28103677/inversion-of-extender-unit-selectivity-in-the-erythromycin-polyketide-synthase-by-acyltransferase-domain-engineering
#3
Irina Koryakina, Christian Kasey, John B McArthur, Andrew N Lowell, Joseph A Chemler, Shasha Li, Douglas A Hansen, David H Sherman, Gavin J Williams
Acyltransferase (AT) domains of polyketide synthases (PKSs) select extender units for incorporation into polyketides and dictate large portions of the structures of clinically relevant natural products. Accordingly, there is significant interest in engineering the substrate specificity of PKS ATs in order to site-selectively manipulate polyketide structure. However, previous attempts to engineer ATs have yielded mutant PKSs with relaxed extender unit specificity, rather than an inversion of selectivity from one substrate to another...
January 20, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28078712/comparative-study-on-fatty-acid-composition-of-olive-olea-europaea-l-with-emphasis-on-phytosterol-contents
#4
Ali Ozkan, Hassan Y Aboul-Enein, Muhittin Kulak, Recep Bindak
The present study was designed to determine the fatty acid composition and phytosterol contents of Turkish native olive cultivars namely (Kilis Yağlık and Nizip Yağlık cv.). In this context, olive fruits from thirty-four locations were sampled and then screened for their components in comparison. Fifteen different fatty acids were found in both olive oils. In the order of abundance, the most important ones were: oleic acid (18:1) > palmitic acid (16:0) > linoleic acid (18:2) > stearic acid (18:0)...
January 12, 2017: Biomedical Chromatography: BMC
https://www.readbyqxmd.com/read/28076344/qualitative-aspects-of-the-process-of-neonatal-hearing-screening-program-in-mexico-evaluated-from-the-parental-perspective
#5
Maria Del Consuelo Martínez Wbaldo, María de Los Ángeles Torres Lagunas
INTRODUCTION: The Universal Newborn Hearing Screening Program in Mexico began in 2010. Its results, published in 2013 by the National Council for the Development and Inclusion of Persons with Disabilities (CONADIS), report low coverage and, currently, there is a dearth of information about its activities. This study describes the process of the program from the epistemological perspective of women whose children participated in the program, evaluating it under the sustenance of the constructivist-respondent model in search of aspects that could help explain its results...
December 12, 2016: Medwave
https://www.readbyqxmd.com/read/28074221/exploring-the-thermostable-properties-of-halohydrin-dehalogenase-from-agrobacterium-radiobacter-ad1-by-a-combinatorial-directed-evolution-strategy
#6
Zhiyun Wu, Wenfeng Deng, Yapei Tong, Qian Liao, Dongmin Xin, Huashun Yu, Juan Feng, Lixia Tang
As a crucial factor for biocatalysts, protein thermostability often arises from a combination of factors that are often difficult to rationalize. In this work, the thermostable nature of halohydrin dehalogenase from Agrobacterium radiobacter AD1 (HheC) was systematically explored using a combinatorial directed evolution approach. For this, a mutagenesis library of HheC mutants was first constructed using error-prone PCR with low mutagenesis frequency. After screening approximately 2000 colonies, six mutants with eight mutation sites were obtained...
January 10, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28072719/association-between-dietary-fat-intake-and-colorectal-adenoma-in-korean-adults-a-cross-sectional-study
#7
Jeehyun Kim, Seung-Won Oh, Young-Sun Kim, Hyuktae Kwon, Hee-Kyung Joh, Ji-Eun Lee, Danbee Park, Jae-Hong Park, Ah-Ryoung Ko, Ye-Ji Kim
The incidence of colorectal cancer is rapidly increasing in South Korea. It is important to clarify the association between colorectal cancer and diet, being one of the main modifiable risk factors, as such studies in the Korean population are lacking.A cross-sectional study was performed using data from participants who had undergone a screening colonoscopy and a nutritional assessment during a routine health check-up from January 2008 to December 2011. Dietary intake data were derived from 1-day food records; colorectal adenoma was histopathologically confirmed by biopsy during colonoscopy...
January 2017: Medicine (Baltimore)
https://www.readbyqxmd.com/read/28064156/high-prevalence-of-baffle-leaks-in-adults-after-atrial-switch-operations-for-transposition-of-the-great-arteries
#8
Gabriella De Pasquale, Francesca Bonassin Tempesta, Bruno Santos Lopes, Daniela Babic, Angela Oxenius, Theresa Seeliger, Christiane Gruner, Felix C Tanner, Patric Biaggi, Christine Attenhofer Jost, Matthias Greutmann
AIMS: To determine the prevalence of baffle leaks in adults after atrial switch operations for transposition of the great arteries, as these may predispose to paradoxical embolic events, particularly in patients with transvenous pacemaker or defibrillator leads. METHODS AND RESULTS: We routinely perform contrast echocardiography with agitated saline in all patients after atrial switch operations. For this study, we analysed patients who had saline contrast echocardiography between 2010 and 2012...
January 6, 2017: European Heart Journal Cardiovascular Imaging
https://www.readbyqxmd.com/read/28060953/what-do-men-want-from-a-health-screening-mobile-app-a-qualitative-study
#9
Chin Hai Teo, Chirk Jenn Ng, Alan White
There is a lack of mobile app which aims to improve health screening uptake developed for men. As part of the study to develop an effective mobile app to increase health screening uptake in men, we conducted a needs assessment to find out what do men want from a health screening mobile app. In-depth interviews and focus group discussions were conducted with 31 men from a banking institution in Kuala Lumpur. The participants were purposely sampled according to their job position, age, ethnicity and screening status...
2017: PloS One
https://www.readbyqxmd.com/read/28044007/modification-of-the-peroxygenative-peroxidative-activity-ratio-in-the-unspecific-peroxygenase-from-agrocybe-aegerita-by-structure-guided-evolution
#10
Diana M Mate, Miguel A Palomino, Patricia Molina-Espeja, Javier Martin-Diaz, Miguel Alcalde
Unspecific peroxygenase (UPO) is a heme-thiolate peroxidase capable of performing with high-selectivity C-H oxyfunctionalizations of great interest in organic synthesis through its peroxygenative activity. However, the convergence of such activity with an unwanted peroxidative activity encumbers practical applications. In this study, we have modified the peroxygenative:peroxidative activity ratio (P:p ratio) of UPO from Agrocybe aegerita by structure-guided evolution. Several flexible loops (Glu1-Pro35, Gly103-Asp131, Ser226-Gly243, Gln254-Thr276 and Ty293-Arg327) were selected on the basis on their B-factors and ΔΔG values...
January 1, 2017: Protein Engineering, Design & Selection: PEDS
https://www.readbyqxmd.com/read/28043739/canadian-cardiovascular-society-canadian-pediatric-cardiology-association-position-statement-on-pulse-oximetry-screening-in-newborns-to-enhance-detection-of-critical-congenital-heart-disease
#11
Kenny K Wong, Anne Fournier, Deborah S Fruitman, Lisa Graves, Derek G Human, Michael Narvey, Jennifer L Russell
Congenital heart disease is the most common congenital malformation and approximately 3 in 1000 newborns have critical congenital heart disease (CCHD). Timely diagnosis affects morbidity, mortality, and disability, and newborn pulse oximetry screening has been studied to enhance detection of CCHD. In this position statement we present an evaluation of the literature for pulse oximetry screening. Current detection strategies including prenatal ultrasound examination and newborn physical examination are limited by low diagnostic sensitivity...
October 26, 2016: Canadian Journal of Cardiology
https://www.readbyqxmd.com/read/28042556/deep-vein-thrombosis-in-patients-with-pulmonary-embolism-prevalance-clinical-significance-and-outcome
#12
Jun Sung Lee, Tong Moon, Tae Hoon Kim, Se Young Kim, Jun Young Choi, Kyung Bok Lee, Yu Jin Kwon, Suk Hee Song, Su Hyun Kim, Hae Ok Kim, Ho Kyeong Hwang, Min Ji Kim, Young Kyoung Lee
PURPOSE: Deep venous thrombosis (DVT) and pulmonary embolism (PE) are considered as similar disease entities representing different clinical manifestations. The objectives of this study were: 1) to determine the prevalence and outcome of DVT in patients with PE; 2) to identify additional risk factors for PE-related unfavorable outcome and 30-day all-cause mortality; and 3) to establish the clinical importance of screening for concomitant DVT. MATERIALS AND METHODS: From January 2013 to December 2015, a total of 141 patients with confirmed PE were evaluated...
December 2016: Vascular Specialist International
https://www.readbyqxmd.com/read/28042234/diagnosis-and-treatment-of-anemia-in-patients-with-inflammatory-bowel-disease
#13
REVIEW
Victoria Mücke, Marcus M Mücke, Tim Raine, Dominik Bettenworth
Anemia represents one of the most frequent complications in inflammatory bowel disease (IBD) and severely impairs the quality of life of affected patients. The etiology of anemia in IBD patients can be multifactorial, often involving a combination of iron deficiency (ID) and anemia of chronic disease (ACD). Although current guidelines recommend screening for and treatment of anemia in IBD patients, current observational data suggest that it still remains underdiagnosed and undertreated. Besides basic laboratory parameters (e...
2017: Annals of Gastroenterology: Quarterly Publication of the Hellenic Society of Gastroenterology
https://www.readbyqxmd.com/read/28040465/toll-like-receptor-2-promiscuity-is-responsible-for-the-immunostimulatory-activity-of-nucleic-acid-nanocarriers
#14
Malvina Pizzuto, Monique Gangloff, Daniel Scherman, Nicholas J Gay, Virginie Escriou, Jean-Marie Ruysschaert, Caroline Lonez
Lipopolyamines (LPAs) are cationic lipids; they interact spontaneously with nucleic acids to form lipoplexes used for gene delivery. The main hurdle to using lipoplexes in gene therapy lies in their immunostimulatory properties, so far attributed to the nucleic acid cargo, while cationic lipids were considered as inert to the immune system. Here we demonstrate for the first time that di-C18 LPAs trigger pro-inflammatory responses through Toll-like receptor 2 (TLR2) activation, and this whether they are bound to nucleic acids or not...
December 28, 2016: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28035629/cost-effectiveness-of-different-population-screening-strategies-for-hereditary-haemochromatosis-in-australia
#15
Barbara de Graaff, Amanda Neil, Lei Si, Kwang Chien Yee, Kristy Sanderson, Lyle Gurrin, Andrew J Palmer
INTRODUCTION: Amongst populations of northern European ancestry, HFE-associated haemochromatosis is a common genetic disorder characterised by iron overload. In the absence of treatment, excess iron is stored in parenchymal tissues, causing morbidity and mortality. Population screening programmes may increase early diagnosis and reduce associated disease. No contemporary health economic evaluation has been published for Australia. The objective of this study was to identify cost-effective screening strategies for haemochromatosis in the Australian setting...
December 29, 2016: Applied Health Economics and Health Policy
https://www.readbyqxmd.com/read/28025326/dysfunction-of-cgmp-signalling-in-photoreceptors-by-a-macular-dystrophy-related-mutation-in-the-calcium-sensor-gcap1
#16
Farina Vocke, Nicole Weisschuh, Valerio Marino, Silvia Malfatti, Samuel G Jacobson, Charlotte M Reiff, Daniele Dell'Orco, Karl-Wilhelm Koch
Macular dystrophy leads to progressive loss of central vision and shows symptoms similar to age-related macular degeneration. Genetic screening of patients diagnosed with macular dystrophy disclosed a novel mutation in the GUCA1A gene, namely a c.526C > T substitution leading to the amino acid substitution p.L176F in the guanylate cyclase-activating protein 1 (GCAP1). The same variant was found in three families showing an autosomal dominant mode of inheritance. For a full functional characterization of the L176F mutant we expressed and purified the mutant protein and measured key parameters of its activating properties, its Ca(2+)/Mg(2+)-binding, and its Ca(2+)-induced conformational changes in comparison to the wildtype protein...
December 26, 2016: Human Molecular Genetics
https://www.readbyqxmd.com/read/28005308/second-generation-engineering-of-thermostable-transketolase-tkgst-for-aliphatic-aldehyde-acceptors-with-improved-and-reversed-stereoselectivity
#17
Chaoqiang Zhou, Thangavelu Saravanan, Marion Lorillière, Dongzhi Wei, Franck Charmantray, Laurence Hecquet, Wolf-Dieter Fessner, Dong Yi
Transketolase from Geobacillus stearothermophilus (TKgst) is a novel thermostable enzyme with notable high activity and stability at elevated temperatures, but accepts non-α-hydroxylated aldehydes only with low efficiency. Here, we report on a protein engineering study by double-site saturation mutagenesis at either Leu191 or Phe435 in combination with Asp470 of TKgst, which are responsible for substrate binding in the active site. Screening of the mutagenesis libraries resulted in several positive variants with up to 7...
December 22, 2016: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28004808/il-13-is-a-therapeutic-target-in-radiation-lung-injury
#18
Su I Chung, Jason A Horton, Thirumalai R Ramalingam, Ayla O White, Eun Joo Chung, Kathryn E Hudak, Bradley T Scroggins, Joseph R Arron, Thomas A Wynn, Deborah E Citrin
Pulmonary fibrosis is a potentially lethal late adverse event of thoracic irradiation. Prior research indicates that unrestrained TGF-β1 and/or type 2 cytokine-driven immune responses promote fibrosis following radiation injury, but the full spectrum of factors governing this pathology remains unclear. Interleukin 13 (IL-13) is a key factor in fibrotic disease associated with helminth infection, but it is unclear whether it plays a similar role in radiation-induced lung fibrosis. Using a mouse model, we tested the hypothesis that IL-13 drives the progression of radiation-induced pulmonary fibrosis...
December 22, 2016: Scientific Reports
https://www.readbyqxmd.com/read/28003128/structural-and-binding-studies-of-a-c-type-galactose-binding-lectin-from-bothrops-jararacussu-snake-venom
#19
Marco A Sartim, Matheus P Pinheiro, Ricardo A P de Pádua, Suely V Sampaio, M Cristina Nonato
BJcuL is a snake venom galactoside-binding lectin (SVgalL) isolated from Bothrops jararacussu and is involved in a wide variety of biological activities including triggering of pro-inflammatory response, disruption of microbial biofilm structure and induction of apoptosis. In the present work, we determined the crystallographic structure of BJcuL, the first holo structure of a SVgalL, and introduced the fluorescence-based thermal stability assay (Thermofluor) as a tool for screening and characterization of the binding mechanism of SVgalL ligands...
December 19, 2016: Toxicon: Official Journal of the International Society on Toxinology
https://www.readbyqxmd.com/read/28000046/semi-rational-engineering-of-leucine-dehydrogenase-for-l-2-aminobutyric-acid-production
#20
Jian-Miao Xu, Feng Cheng, Fang-Tian Fu, Hai-Feng Hu, Yu-Guo Zheng
L-2-aminobutyric acid (L-ABA) as a precursor for the anticonvulsant and the antituberculotic is a key intermediate in the chemical and pharmaceutical industries. Recently, leucine dehydrogenase (LeuDH) with NAD(+) regeneration was developed for L-ABA production on a large scale. Previously, the L-ABA yield was improved by optimizing conversion conditions, including cofactor regeneration and enzyme immobilization but not protein engineering on LeuDH due to lacking an applicable high-throughput screening (HTS) method...
December 20, 2016: Applied Biochemistry and Biotechnology
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