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https://www.readbyqxmd.com/read/28220435/rapid-construction-of-multiplexed-crispr-cas9-systems-for-plant-genome-editing
#1
Levi Lowder, Aimee Malzahn, Yiping Qi
Multiplex CRISPR-Cas9 nuclease mediated genome editing is an efficient method for disrupting gene function in plants. Use of CRISPR-Cas9 has escalated rapidly in recent years and is expected to become routine practice in molecular biology and related fields of research. Due to the relatively novel and widespread adoption of this technology, first-time users may not have regular access to experienced guidance or technical support from peers or mentors. Here, we offer guidance and technical support in the form of a detailed and tested protocol for simultaneous targeting of three separate loci on the TRANSPARENT TESTA 4 (TT4) gene in Arabidopsis thaliana using multiplex CRISPR-Cas9...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28219684/development-of-magnetic-nanoparticle-based-calorimetric-assay-for-the-detection-of-bovine-mastitis-in-cow-milk
#2
Raja Chinnappan, Sana Al Attas, Gerrit Koop, Tine van Werven, Wendy E Kaman, Floris J Bikker, Mohammed Zourob
Mastitis in dairy cattle is an inflammatory reaction of the udder tissue. Mastitis increases plasmin levels, leading to an increased proteolysis of milk proteins such as casein, resulting in a significant decrease in milk quality and related dairy products. Due to its key-role in mastitis, we used plasmin proteolytic activity as a biomarker for the detection of mastitis in bovine mastitic milk. Inspired by earlier studies on protease activity using mastitic milk samples, we developed a simple colorimetric assay to distinguish mastitic milk from milk derived from healthy animals...
February 17, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28218837/rapid-and-efficient-genome-editing-in-staphylococcus-aureus-by-using-an-engineered-crispr-cas9-system
#3
Weizhong Chen, Yifei Zhang, Won-Sik Yeo, Taeok Bae, Quanjiang Ji
Staphylococcus aureus, a major human pathogen, has been the cause of serious infectious diseases with a high mortality rate. Although genetics is a key means to study S. aureus physiology, such as drug resistance and pathogenesis, genetic manipulation in S. aureus is always time consuming and labor intensive. Here, we report a CRISPR/Cas9 system (pCasSA) for rapid and efficient genome editing, including gene deletion, insertion and single-base substitution mutation in S. aureus. The designed pCasSA system is amenable to assembly of spacers and repair arms by Golden Gate assembly and Gibson assembly, respectively, enabling rapid construction of the plasmids for editing...
February 20, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28217858/golden-gate-assembly-system-dedicated-to-complex-pathway-manipulation-in-yarrowia-lipolytica
#4
Ewelina Celińska, Rodrigo Ledesma-Amaro, Macarena Larroude, Tristan Rossignol, Cyrille Pauthenier, Jean-Marc Nicaud
In this study, we have adopted Golden Gate modular cloning strategy to develop a robust and versatile DNA assembly platform for the nonconventional yeast Yarrowia lipolytica. To this end, a broad set of destination vectors and interchangeable building blocks have been constructed. The DNA modules were assembled on a scaffold of predesigned 4 nt overhangs covering three transcription units (each bearing promoter, gene and terminator), selection marker gene and genomic integration targeting sequences, constituting altogether thirteen elements...
February 19, 2017: Microbial Biotechnology
https://www.readbyqxmd.com/read/28217534/assessment-of-facial-golden-proportions-among-central-indian-population
#5
Rathore Saurabh, Bolya Piyush, Bhatt Sourabh, Ojha Preeti, Rutvik Trivedi, Pradeep Vishnoi
AIM: This study aimed to identify and establish the facial and smile proportions in young adults and to compare the results with ideal or divine proportions, compare the proportions of males and females included in our study population and compare them with those established for Caucasian and Japanese populations. MATERIALS AND METHODS: Two hundred participants (164 females, 36 males) with Angle's class I malocclusion (M.O). and well-balanced faces were selected and photographed in the frontal repose position...
December 2016: Journal of International Society of Preventive & Community Dentistry
https://www.readbyqxmd.com/read/28216927/intraprostatic-ozone-therapy-a-minimally-invasive-approach-in-benign-prostatic-hyperplasia
#6
Shabbir Hussain, Deepti B Sharma, Fanindra S Solanki, Ajay Pathak, Dhananjay Sharma
INTRODUCTION: Transurethral resection of prostate (TURP) remains the golden standard therapy since decades. There are various minimally invasive therapies (MITs) for the treatment of benign prostatic hyperplasia (BPH). Still, there is a need for therapy with lesser side effects and better outcome. We had studied the effect of intraprostatic ozone injection (IPOI) as an MIT for patients with BPH who have failed trial without catheter (TWOC). MATERIALS AND METHODS: Thirty elderly patients with BPH with a prostate size of 30 g or more were enrolled for the study...
January 2017: Urology Annals
https://www.readbyqxmd.com/read/28216038/de-novo-transcriptome-of-the-muga-silkworm-antheraea-assamensis-helfer
#7
Hasnahana Chetia, Debajyoti Kabiraj, Deepika Singh, Ponnala Vimal Mosahari, Suradip Das, Pragya Sharma, Kartik Neog, Swagata Sharma, P Jayaprakash, Utpal Bora
Antheraea assamensis (Lepidoptera: Saturniidae), is a semi-domesticated silkworm known to be endemic to Assam and the adjoining hilly areas of Northeast India. It is the only producer of a unique, commercially important variety of golden silk called "muga silk". Herein, we report the de novo transcriptome of A. assamensis reared on Machilus bombycina leaves for the first time. Short reads generated by high throughput sequencing of cDNA libraries from multiple tissues, viz. alimentary canal, silk gland and residual body were assembled into transcripts via a de novo assembly pipeline followed by functional annotation and classification...
February 16, 2017: Gene
https://www.readbyqxmd.com/read/28215683/effects-of-hormonal-stimulation-on-the-concentration-and-quality-of-excreted-spermatozoa-in-the-critically-endangered-panamanian-golden-frog-atelopus-zeteki
#8
Gina Della Togna, Vance L Trudeau, Brian Gratwicke, Matthew Evans, Lauren Augustine, Han Chia, Edward J Bronikowski, James B Murphy, Pierre Comizzoli
Knowledge of basic gamete biology is critical to better protect and propagate endangered amphibian species and also to develop reproductive technologies combined with germplasm cryopreservation. The objectives of the study were to test different hormonal stimulations and then characterize the quantity and quality of Panamanian golden frog (Atelopus zeteki) spermatozoa. Following intraperitoneal injection of the gonadotropin-releasing hormone agonist (des-Gly(10), D-Ala(6), Pro-NHEt(9-)-GnRH 1, 2 or 4 μg/g of body weight), human chorionic gonadotropin (hCG; 5 or 10 IU/gbw), or Amphiplex™ (0...
March 15, 2017: Theriogenology
https://www.readbyqxmd.com/read/28204911/selection-of-transformation-efficient-barley-genotypes-based-on-tfa-transformation-amenability-haplotype-and-higher-resolution-mapping-of-the-tfa-loci
#9
Hiroshi Hisano, Brigid Meints, Matthew J Moscou, Luis Cistue, Begoña Echávarri, Kazuhiro Sato, Patrick M Hayes
The genetic substitution of transformation amenability alleles from 'Golden Promise' can facilitate the development of transformation-efficient lines from recalcitrant barley cultivars. Barley (Hordeum vulgare) cv. 'Golden Promise' is one of the most useful and well-studied cultivars for genetic manipulation. In a previous report, we identified several transformation amenability (TFA) loci responsible for Agrobacterium-mediated transformation using the F2 generation of immature embryos, derived from 'Haruna Nijo' × 'Golden Promise,' as explants...
February 15, 2017: Plant Cell Reports
https://www.readbyqxmd.com/read/28201891/electron-transport-through-a-spin-crossover-junction-perspectives-from-a-wavefunction-based-approach
#10
Sergi Vela, Martin Verot, Emmanuel Fromager, Vincent Robert
The present paper reports the application of a computational framework, based on the quantum master equation, the Fermi's golden Rule, and conventional wavefunction-based methods, to describe electron transport through a spin crossover molecular junction (Fe(bapbpy) (NCS)2, 1, bapbpy = N-(6-(6-(Pyridin-2-ylamino)pyridin-2-yl)pyridin-2-yl)-pyridin-2-amine). This scheme is an alternative to the standard approaches based on the relative position and nature of the frontier orbitals, as it evaluates the junction's Green's function by means of accurate state energies and wavefunctions...
February 14, 2017: Journal of Chemical Physics
https://www.readbyqxmd.com/read/28199338/development-and-application-of-an-edna-method-to-detect-the-critically-endangered-trinidad-golden-tree-frog-phytotriades-auratus-in-bromeliad-phytotelmata
#11
Sarah Brozio, Chloe Manson, Eleanor Gourevitch, Thomas J Burns, Mark S Greener, J Roger Downie, Paul A Hoskisson
The use of environmental DNA (eDNA) to monitor rare and elusive species has great potential for conservation biology. Traditional surveying methods can be time-consuming, labour-intensive, subject to error or can be invasive and potentially damaging to habitat. The Trinidad golden treefrog (Phytotriades auratus) is one such species that would benefit from such an approach. This species inhabits the giant bromeliad (Glomeropitcairnia erectiflora) on two peaks on the Caribbean island of Trinidad. Traditional survey methods for this species have required the destruction of the giant bromeliad, which is the only known habitat of this frog...
2017: PloS One
https://www.readbyqxmd.com/read/28196365/the-golden-age-of-probiotics-a-systematic-review-and-meta-analysis-of-randomized-and-observational-studies-in-preterm-infants
#12
Elda Dermyshi, Yizhong Wang, Chongbin Yan, Wenchao Hong, Gang Qiu, Xiaohui Gong, Ting Zhang
BACKGROUND: Over the last few years, probiotics have been one of the most studied interventions in neonatal medicine. OBJECTIVES: The aim of this work was to analyse all studies (randomized controlled trials, RCTs, and observational studies) assessing the use of probiotics in very low birth weight (VLBW) preterm infants. SEARCH METHODS: A systematic literature search was conducted using PubMed, Embase, Cochrane Library, and Web of Science...
February 15, 2017: Neonatology
https://www.readbyqxmd.com/read/28195343/galectin-9-diverse-roles-in-hepatic-immune-homeostasis-and-inflammation
#13
REVIEW
Lucy Golden-Mason, Hugo R Rosen
Glycan-binding proteins which include galectins are involved at all stages of immunity and inflammation, ranging from initiation through resolution. Galectin-9 (Gal-9) is highly expressed in the liver and has a wide variety of biological functions in innate and adaptive immunity that are instrumental in the maintenance of hepatic homeostasis. In the setting of viral hepatitis, increased expression of Gal-9 drives the expansion of regulatory T cells and contraction of effector T cells, thereby favoring viral persistence...
February 13, 2017: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
https://www.readbyqxmd.com/read/28195150/molecular-dietary-analysis-of-two-sympatric-felids-in-the-mountains-of-southwest-china-biodiversity-hotspot-and-conservation-implications
#14
Mengyin Xiong, Dajun Wang, Hongliang Bu, Xinning Shao, Dan Zhang, Sheng Li, Rongjiang Wang, Meng Yao
Dietary information is lacking in most of small to mid-sized carnivores due to their elusive predatory behaviour and versatile feeding habits. The leopard cat (LPC; Prionailurus bengalensis) and the Asiatic golden cat (AGC; Catopuma temminckii) are two important yet increasingly endangered carnivore species in the temperate mountain forest ecosystem in Southwest China, a global biodiversity hotspot and a significant reservoir of China's endemic species. We investigated the vertebrate prey of the two sympatric felids using faecal DNA and a next-generation sequencing (NGS)/metabarcoding approach...
February 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28194481/accumulation-of-metals-in-liver-tissues-of-sympatric-golden-jackal-canis-aureus-and-red-fox-vulpes-vulpes-in-the-southern-part-of-romania
#15
Attila Farkas, András Bidló, Bernadett Bolodár-Varga, Ferenc Jánoska
Several previous study results have already demonstrated that golden jackal and red fox may serve as biological indicators of trace elements and heavy metal concentrations in the various regions they inhabit. The aim of this study was to evaluate accumulation patterns of targeted elements (Al, Ca, Cr, Cu, Fe, Mg, Mn, Ni and Pb) in liver samples of red foxes and golden jackals collected during the same period in the southern part of Romania. The accumulation patterns of trace elements in the livers of sympatric golden jackal and red fox were practically the same...
February 13, 2017: Bulletin of Environmental Contamination and Toxicology
https://www.readbyqxmd.com/read/28193216/a-high-throughput-expression-screening-platform-to-optimize-the-production-of-antimicrobial-peptides
#16
Christine Schreiber, Hagen Müller, Oliver Birrenbach, Moritz Klein, Doreen Heerd, Tobias Weidner, Denise Salzig, Peter Czermak
BACKGROUND: Antimicrobial peptides (AMPs) are promising candidates for the development of novel antibiotics, but it is difficult to produce sufficient quantities for preclinical and clinical studies due to their toxicity towards microbial expression hosts. To avoid laborious trial-and-error testing for the identification of suitable expression constructs, we have developed a small-scale expression screening platform based on a combinatorial plasmid library. RESULTS: The combinatorial library is based on the Golden Gate cloning system...
February 13, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28188424/fine-tuning-of-the-flavonoid-and-monolignol-pathways-during-apple-early-fruit-development
#17
Paolo Baldi, Mirko Moser, Matteo Brilli, Urska Vrhovsek, Massimo Pindo, Azeddine Si-Ammour
A coordinated regulation of different branches of the flavonoid pathway was highlighted that may contribute to elucidate the role of this important class of compounds during the early stages of apple fruit development. Apple (Malus × domestica Borkh.) is an economically important fruit appreciated for its organoleptic characteristics and its benefits for human health. The first stages after fruit set represent a very important and still poorly characterized developmental process. To enable the profiling of genes involved in apple early fruit development, we combined the suppression subtractive hybridization (SSH) protocol to next-generation sequencing...
February 11, 2017: Planta
https://www.readbyqxmd.com/read/28187938/occurrence-of-veterinary-pharmaceuticals-in-golden-eagle-nestlings-unnoticed-scavenging-on-livestock-carcasses-and-other-potential-exposure-routes
#18
Guillermo Blanco, Alexandra Junza, Dolores Barrón
Wildlife exposure to pharmaceuticals can occur through contaminated water, and through the excreta and carcasses of medicated livestock, with potential for bioaccumulation and transfer through food webs. We evaluated whether nestling exposure to pharmaceuticals can occur from food delivered to nests in the golden eagle (Aquila chrysaetos), a top predator and facultative scavenger. Despite the fact that diet analysis suggests an apparently low dependence on livestock carcasses reduced to two piglets remains (1...
February 7, 2017: Science of the Total Environment
https://www.readbyqxmd.com/read/28187760/case-report-subacute-tetraplegia-in-an-immunocompromised-patient
#19
Daniel Zeller, Anke Heidemeier, Götz Ulrich Grigoleit, Wolfgang Müllges
BACKGROUND: Clinical reasoning in Neurology is based on general associations which help to deduce the site of the lesion. However, even "golden principles" may occasionally be deceptive. Here, we describe the case of subacute flaccid tetraparesis due to motor cortical lesions. To our knowledge, this is the first report to include an impressive illustration of nearly symmetric motor cortical involvement of encephalitis on brain MRI. CASE PRESENTATION: A 51 year old immunocompromized man developed a high-grade pure motor flaccid tetraparesis over few days...
February 10, 2017: BMC Neurology
https://www.readbyqxmd.com/read/28187551/systematics-of-a-small-i-gehyra-i-squamata-gekkonidae-from-the-einasleigh-uplands-queensland-description-of-a-new-range-restricted-species
#20
Gayleen Bourke, Renae C Pratt, Eric Vanderduys, Craig Moritz
The tropical savannah landscapes of Australia's north, though previously overshadowed by the biodiverse rainforests of the Wet Tropics, are themselves now attracting interest for their biological significance and uniqueness. The Einasleigh Uplands region of north-east Queensland is home to a diverse group of mammals and reptiles and was previously recognised for its importance in shaping speciation in birds. Here we add a small saxicoline gecko to a growing list of recently described reptiles that are endemic to this region...
February 9, 2017: Zootaxa
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