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https://www.readbyqxmd.com/read/28193316/polydatin-improves-the-developmental-competence-of-bovine-embryos-in-vitro-via-induction-of-sirtuin-1-sirt1
#1
Imran Khan, Sung Woo Kim, Kyung-Lim Lee, Seok-Hwan Song, Ayman Mesalam, M M R Chowdhury, Zia Uddin, Ki Hun Park, Il-Keun Kong
The aim of the present study was to investigate the beneficial effect of polydatin (PD), the glycoside form of resveratrol, on embryo development in vitro. Oocytes were aspirated from ovaries of Korean Hanwoo cows and cultured until Day 8 in a humidified atmosphere of 5% CO2 in air at 38.5°C. Protein and gene expression levels were determined through confocal microscopy and reverse transcription-polymerase chain reaction respectively, whereas the number of total and apoptotic cells in Day 8 blastocysts was determined using Hoechst 33342 staining and terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling...
February 14, 2017: Reproduction, Fertility, and Development
https://www.readbyqxmd.com/read/28174075/molecular-detection-of-genotype-ii-grass-carp-reovirus-based-on-nucleic-acid-sequence-based-amplification-combined-with-enzyme-linked-immunosorbent-assay-nasba-elisa
#2
Weiwei Zeng, Wei Yao, Yingying Wang, Yingying Li, Sven M Bermann, Yan Ren, Cunbin Shi, Xinjian Song, Qiwen Huang, Shuchen Zheng, Qing Wang
Grass carp reovirus (GCRV) is the causative agent of the grass carp hemorrhagic disease that has resulted in severe economic losses in the grass carp (Ctenopharyngodon idella) farming industry in China. Early diagnosis and vaccine administration are important priorities for GCRV control. In this study, a nucleic acid sequence-based amplification with enzyme-linked immunosorbent assay (NASBA-ELISA) was developed for to detect genotype II GCRV (GCRV- II). Primers specifically targeting viral RNA genome segment 6 were utilized for amplification in an isothermal digoxigenin-labeling NASBA process, resulting in DIG-labeled RNA amplicons...
February 4, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28133587/non-radioactive-trf-assay-modifications-to-improve-telomeric-dna-detection-efficiency-in-plants
#3
Liliia R Nigmatullina, Margarita R Sharipova, Eugene V Shakirov
The length of telomeric DNA is often considered a cellular biomarker of aging and general health status. Several telomere length measuring assays have been developed, of which the most common is the Telomere Restriction Fragment (TRF) analysis, which typically involves the use of radioactively labeled oligonucleotide probes. While highly effective, this method potentially poses substantial health concerns and generates radioactive waste. Digoxigenin (DIG) alternatives to radioactive probes have been developed and used successfully in a number of assays...
December 2016: BioNanoScience
https://www.readbyqxmd.com/read/28043537/usefulness-of-is6110-based-restriction-fragment-length-polymorphism-analysis-in-fingerprinting-of-mycobacterium-tuberculosis-isolates-in-north-india
#4
Jitendra Prasad Mathuria, Shampa Anupurba
OBJECTIVE/BACKGROUND: World Health Organization estimates that approximately one-third of the global community is infected with Mycobatcerium tuberculosis (MTB). Various molecular epidemiology methods were developed and found very promising for assessing the genetic diversity among MTB complex strain. The two major tools restriction fragment length polymorphism (RFLP) and spoligotyping were commonly used in various studies. Some Indian studies raise the question about the utility of IS6110-RFLP in India due to the presence of zero and low copy number of IS6110 element in MTB isolates...
December 2016: International Journal of Mycobacteriology
https://www.readbyqxmd.com/read/28007594/the-p20r-mutation-of-%C3%AE-b-crystallin-diminishes-its-anti-apoptotic-activity-in-human-lens-epithelial-cells
#5
Peiran Zhu, Weiwei Li, Mengxia Ni, Cui Zhang, Shuaimei Liu, Qiuyue Wu, Weijun Jiang, Jing Zhang, Mingchao Zhang, Xiaojun Li, Yingxia Cui, Chunyan Xue, Xinyi Xia
αB-crystallin acts as an anti-apoptosis protein in human lens epithelial (HLE) cells. We recently identified a missense mutation in αB-crystallin that changes proline 20 to an arginine (P20R) in a Chinese family with autosomal dominant congenital posterior polar cataract. The impact of the P20R mutation on the anti-apoptosis function remains unclear. To explore the anti-apoptotic activity of αB-crystallin wild type (αB-wt) and its P20R mutant under oxidative stress, HLE cells were transfected with αB-wt and αB-P20R constructs and expression was measured by western blotting...
January 29, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28003732/regulation-of-photoreceptor-gene-transcription-via-a-highly-conserved-transcriptional-regulatory-element-by-vsx-gene-products
#6
Yi Pan, Daniel F Comiskey, Lisa E Kelly, Dawn S Chandler, Heithem M El-Hodiri
PURPOSE: The photoreceptor conserved element-1 (PCE-1) sequence is found in the transcriptional regulatory regions of many genes expressed in photoreceptors. The retinal homeobox (Rx or Rax) gene product functions by binding to PCE-1 sites. However, other transcriptional regulators have also been reported to bind to PCE-1. One of these, vsx2, is expressed in retinal progenitor and bipolar cells. The purpose of this study is to identify Xenopus laevis vsx gene products and characterize vsx gene product expression and function with respect to the PCE-1 site...
2016: Molecular Vision
https://www.readbyqxmd.com/read/27929598/sclerotic-effect-of-oxytetracycline-on-the-submandibular-gland-an-experimental-model
#7
Oğuz Güçlü, Asli Muratli, Deniz Arik, Kazım Tekin, Halil Erdogan, Fevzi Sefa Dereköy
Oxytetracycline has been suggested as an alternate therapy for chronic recurrent sialadenitis and sialorrhea. We conducted an experimental study to investigate the sclerotic effect of this drug on the submandibular gland by histopathologic methods. Our subjects were 20 New Zealand white rabbits, which were divided into two groups of 10. The right submandibular gland of the rabbits in the active-treatment group was injected with 0.3 ml of oxytetracycline (100 mg/ml), and that of the controls was injected with saline...
December 2016: Ear, Nose, & Throat Journal
https://www.readbyqxmd.com/read/27825888/sdr-elisa-ultrasensitive-and-high-throughput-nucleic-acid-detection-based-on-antibody-like-dna-nanostructure
#8
Junlin Wen, Junhua Chen, Li Zhuang, Shungui Zhou
An ultrasensitive and high-throughput nucleic acid detection system, termed as strand displacement reaction-enzyme linked immunosorbent assay (SDR-ELISA), has been developed on the basis of antibody-like DNA nanostructures. Three digoxigenin or biotin modified hairpin probes are utilized to construct antibody-like DNA nanostructures that feature affinity toward streptavidin and anti-digoxigenin antibody via isothermal target-triggered SDR amplification. These antibody-like nanostructures have been employed to conjugate horseradish-peroxidase-labeled anti-digoxigenin antibody with streptavidin that is immobilized on microliter plate wells for enzyme-linked colorimetric assay...
April 15, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27717773/development-and-evaluation-of-molecular-methods-for-detection-of-cryptosporidium-spp-in-human-clinical-samples
#9
Shehla Khalil, Bijay Ranjan Mirdha, Jaishree Paul, Ashutosh Panda, Govind Makharia, Rama Chaudhry, Shinjini Bhatnagar
Cryptosporidiosis is predominantly a gastrointestinal disease of humans and other animals, caused by various species of protozoan parasites representing the genus Cryptosporidium. Detection of Cryptosporidium spp. in human clinical samples is central to the prevention, surveillance and control of cryptosporidiosis, particularly given that there is presently no broadly applicable treatment regimen for this disease. A non-radioactive, genus specific DNA dot blot hybridization assay was developed using Digoxigenin (DIG) labelled probes to detect Cryptosporidium DNA in human clinical samples...
November 2016: Experimental Parasitology
https://www.readbyqxmd.com/read/27714704/development-and-evaluation-of-enzyme-linked-immunosorbent-assay-of-nucleic-acid-sequence-based-amplification-for-diagnosis-of-invasive-aspergillosis
#10
Li Du, Yun Xia, Yunyan He, Qingquan Pu, Ruoyi Hua, Wenyao Wu
Invasive aspergillosis (IA) is a life-threatening infection in immunocompromised patients, rapid and sensitive detection of Aspergillus from clinical samples has been a major challenge in the early diagnosis of IA. An enzyme-linked immunosorbent assay of nucleic acid sequence-based amplification (NASBA-ELISA) was developed to fulfil the need for the efficient diagnosis of these infections. The primers targeting 18S rRNA were selected for the amplification of Aspergillus RNA by the isothermal digoxigenin (DIG)-labeling NASBA process...
December 2016: AMB Express
https://www.readbyqxmd.com/read/27679415/proliferation-and-apoptosis-processes-in-the-seasonal-testicular-development-of-the-wild-daurian-ground-squirrel-citellus-dauricus-brandt-1844
#11
Yingying Han, Jinqi Zhan, Ying Xu, Fengwei Zhang, Zhengrong Yuan, Qiang Weng
The aim of the present study was to elucidate the regulatory role of cell proliferation and apoptosis in testicular development of wild Daurian ground squirrels during the breeding season (April), the non-breeding season (June) and before hibernation (September). Gross mass and hormonal analysis showed that the testis:body mass ratio and plasma testosterone concentration fluctuated seasonally, with a peak in April and lowest values in June. Similarly, spermatogenesis was fully developed in April but suppressed in June and September...
September 28, 2016: Reproduction, Fertility, and Development
https://www.readbyqxmd.com/read/27600327/localization-of-rem2-in-the-central-nervous-system-of-the-adult-rainbow-trout-oncorhynchus-mykiss
#12
Anna G Downs, Katie R Scholles, David M Hollis
Rem2 is member of the RGK (Rem, Rad, and Gem/Kir) subfamily of the Ras superfamily of GTP binding proteins known to influence Ca(2+) entry into the cell. In addition, Rem2, which is found at high levels in the vertebrate brain, is also implicated in cell proliferation and synapse formation. Though the specific, regional localization of Rem2 in the adult mammalian central nervous system has been well-described, such information is lacking in other vertebrates. Rem2 is involved in neuronal processes where the capacities between adults of different vertebrate classes vary...
December 2016: Journal of Chemical Neuroanatomy
https://www.readbyqxmd.com/read/27599431/comparing-nucleic-acid-lateral-flow-and-electrochemical-genosensing-for-the-simultaneous-detection-of-foodborne-pathogens
#13
A Ben Aissa, J J Jara, R M Sebastián, A Vallribera, S Campoy, M I Pividori
Due to the increasing need of rapid tests for application in low resource settings, WHO summarized their ideal features under the acronym ASSURED (Affordable, Sensitive, Specific, User-friendly, Rapid and Robust, Equipment-free, Delivered to those who need it). In this work, two different platforms for the rapid and simultaneous testing of the foodborne pathogens E. coli O157:H7 and Salmonella enterica, in detail a nucleic acid lateral flow and an electrochemical magneto-genosensor are presented and compared in terms of their analytical performance...
February 15, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27598139/liquid-hybridization-and-solid-phase-detection-a-highly-sensitive-and-accurate-strategy-for-microrna-detection-in-plants-and-animals
#14
Fosheng Li, Lanju Mei, Cheng Zhan, Qiang Mao, Min Yao, Shenghua Wang, Lin Tang, Fang Chen
MicroRNAs (miRNAs) play important roles in nearly every aspect of biology, including physiological, biochemical, developmental and pathological processes. Therefore, a highly sensitive and accurate method of detection of miRNAs has great potential in research on theory and application, such as the clinical approach to medicine, animal and plant production, as well as stress response. Here, we report a strategic method to detect miRNAs from multicellular organisms, which mainly includes liquid hybridization and solid phase detection (LHSPD); it has been verified in various species and is much more sensitive than traditional biotin-labeled Northern blots...
September 1, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27587586/genome-wide-identification-and-characterisation-of-human-dna-replication-origins-by-initiation-site-sequencing-ini-seq
#15
Alexander R Langley, Stefan Gräf, James C Smith, Torsten Krude
Next-generation sequencing has enabled the genome-wide identification of human DNA replication origins. However, different approaches to mapping replication origins, namely (i) sequencing isolated small nascent DNA strands (SNS-seq); (ii) sequencing replication bubbles (bubble-seq) and (iii) sequencing Okazaki fragments (OK-seq), show only limited concordance. To address this controversy, we describe here an independent high-resolution origin mapping technique that we call initiation site sequencing (ini-seq)...
December 1, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27427471/spred-2-expression-is-associated-with-neural-repair-of-injured-adult-zebrafish-brain
#16
Fei Tieng Lim, Satoshi Ogawa, Ishwar S Parhar
Sprouty-related protein-2 (Spred-2) is a negative regulator of extracellular signal-regulated kinases (ERK) pathway, which is important for cell proliferation, neuronal differentiation, plasticity and survival. Nevertheless, its general molecular characteristics such as gene expression patterns and potential role in neural repair in the brain remain unknown. Thus, this study aimed to characterize the expression of spred-2 in the zebrafish brain. Digoxigenin-in situ hybridization showed spred-2 mRNA-expressing cells were mainly seen in the proliferative zones such as the olfactory bulb, telencephalon, optic tectum, cerebellum, and the dorsal and ventral hypothalamus, and most of which were neuronal cells...
July 14, 2016: Journal of Chemical Neuroanatomy
https://www.readbyqxmd.com/read/27369073/ca-2-calmodulin-dependent-protein-kinase-phosphatase-camkp-ppm1f-interacts-with-neurofilament-l-and-inhibits-its%C3%A2-filament-association
#17
Hana Ozaki, Tsuyoshi Katoh, Ryoko Nakagawa, Yasuhiro Ishihara, Noriyuki Sueyoshi, Isamu Kameshita, Takanobu Taniguchi, Tetsuo Hirano, Takeshi Yamazaki, Atsuhiko Ishida
Ca(2+)/calmodulin-dependent protein kinase phosphatase (CaMKP/PPM1F) is a Ser/Thr phosphatase that belongs to the PPM family. Growing evidence suggests that PPM phosphatases including CaMKP act as a complex with other proteins to regulate cellular functions. In this study, using the two-dimensional far-western blotting technique with digoxigenin-labeled CaMKP as a probe, in conjunction with peptide mass fingerprinting analysis, we identified neurofilament L (NFL) as a CaMKP-binding protein in a Triton-insoluble fraction of rat brain...
September 2, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27341462/clinicopathological-analysis-of-mirna-expression-in-breast-cancer-tissues-by-using-mirna-in-situ-hybridization
#18
Yogin Patel, Ji S Lee, Hexin Chen
In this article, we describe a detailed protocol for miRNA detection in breast cancer tissue using in situ hybridization with a digoxigenin-labelled LNA (Locked Nucleic Acid) probe. The probe was recognized by anti-DIG alkaline phosphatase antibodies and later developed using alkaline phosphatase substrate producing fluorescence signals. Here we utilized miRNA in situ hybridization (MISH) technique to analyze expression of miR-489 in tissues from breast cancer patients. This technique can detect the localization of miRNA of interest in individual tissue samples...
2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27286808/generation-of-digoxigenin-incorporated-probes-to-enhance-dna-detection-sensitivity
#19
Tsung-Po Lai, Woodring E Wright, Jerry W Shay
Telomere length in humans has been correlated with cancer and age-related diseases. The standard method to measure telomere length relies on Southern blot analysis with radioactively or non-radioactively labeled probes containing several telomeric DNA repeats. However, this approach requires relatively large amounts of genomic DNA, making it difficult to measure telomere length when a limited amount of sample is available. Here, we describe a non-radioactive labeling method that uses 3' fill-in combined with lambda exonuclease digestion to incorporate one or more digoxigenin (DIG) molecules into bridged nucleic acid (BNA)-containing oligonucleotides (ONTs)...
2016: BioTechniques
https://www.readbyqxmd.com/read/27269788/an-elisa-for-quantifying-quail-igy-and-characterizing-maternal-igy-transfer-to-egg-yolk-in-several-quail-strains
#20
Atsushi Murai, Misako Kakiuchi, Takahito Hamano, Misato Kobayashi, Masaoki Tsudzuki, Mikiharu Nakano, Yoichi Matsuda, Fumihiko Horio
In avian species, maternal blood immunoglobulin Y (IgY) is transferred to the egg yolks of maturing oocytes, but the mechanism underlying this transfer is unknown. To gain insight into the mechanism of maternal IgY transfer in quail, we established an enzyme-linked immunosorbent assay (ELISA) for the quantitation of quail IgY. We characterized strain differences in blood and egg yolk IgY concentrations and exogenously injected IgY-Fc uptakes into egg yolks. A specific rabbit polyclonal antibody to quail IgY was raised for the ELISA...
July 2016: Veterinary Immunology and Immunopathology
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