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https://www.readbyqxmd.com/read/28633895/synthesis-of-meon-neoglycosides-of-digoxigenin-with-6-deoxy-and-2-6-dideoxy-d-glucose-derivatives-and-their-anticancer-activity
#1
Dong-Dong Wang, Xiao-San Li, Yu-Zhou Bao, Jie Liu, Xiao-Kun Zhang, Xin-Sheng Yao, Xue-Long Sun, Jin-Shan Tang
Cardiac glycosides show anticancer activities and their deoxy-sugar chains are vital for their anticancer effects. In order to study the structure-activity relationship (SAR) of cardiac glycosides toward cancers and get more potent anticancer agents, a series of MeON-neoglycosides of digoxigenin was synthesized and evaluated. First, ten 6-deoxy- and 2,6-dideoxy-d-glucopyranosyl donors were synthesized starting from methyl α-d-glucopyranoside and 2-deoxy-d-glucose. Meanwhile, the digoxigenin was obtained by acidic hydrolysis of commercially available digoxin as glycosyl acceptor...
June 3, 2017: Bioorganic & Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/28608344/nonradioactive-detection-of-small-rnas-using-digoxigenin-labeled-probes
#2
Ariel H Tomassi, Delfina Gagliardi, Damian A Cambiagno, Pablo A Manavella
Small RNAs have been traditionally detected and quantified using small RNA blots, a modified Northern blot technique. The small RNAs are size-fractionated from the rest of the cellular RNA molecules by polyacrylamide gel electrophoresis and transferred by blotting onto a positively charged membrane. A radiolabeled probe was then traditionally used to detect a specific small RNA in the cellular pool. Small RNA blotting is a relatively simple, inexpensive approach to visualize small RNAs without artifacts. However, the radioactive labeling of the probe is sometimes an impediment, especially due to the requirement of specialized facilities...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28595095/differential-expression-between-dsp-only-and-dsp-pp523-transcripts-in-rat-molar-teeth
#3
Ya-Qin Zhu, Ryan M Song, Helena H Ritchie
OBJECTIVE: To compare the expression patterns of two multiple transcripts derived from DSP-PP gene during tooth development. One is DSP-only transcript (i.e. does not encode PP) and the other is DSP-PP523 transcript, a main DSP-PP transcript. DESIGN: Unique antisense and sense riboprobes were generated from DSP-only and DSPPP523 cDNAs for in situ studies to examine DSP-only and DSP-PP523 transcript expression in developing molars. Paraffin-embedded sections (5-7μ m) from embryonic 20day, postnatal 2, 3 and 6days were deparaffined and hydrated...
May 31, 2017: Archives of Oral Biology
https://www.readbyqxmd.com/read/28573617/nonradioactive-northern-blot-analysis-to-detect-ebola-virus-minigenomic-mrna
#4
Kristina Brauburger, Tessa Cressey, Elke Mühlberger
In this chapter, we describe the detection of Ebola virus minigenomic mRNA using a nonradioactive Northern hybridization. This protocol comprises all steps beginning with the synthesis of a digoxigenin-labeled riboprobe, harvest of transcribed mRNA from cells transfected with the Ebola virus minigenome system, separation of mRNA species by denaturing RNA gel electrophoresis, transfer of the mRNA to nylon membranes by vacuum blotting, and finally the detection of minigenome-specific mRNA through hybridization with a labeled riboprobe directed against the reporter gene...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28442025/protective-effects-of-polydatin-on-experimental-testicular-torsion-and-detorsion-injury-in-rats
#5
Huilian Qiao, He Ma, Wanjun Cao, Hao Chen, Jinhua Wei, Zhen Li
Oxidative stress plays a critical role in the process of testicular torsion and detorsion (T/D). The purpose of the present study was to investigate the protective effect of polydatin (PD) on testicular T/D injury. Rats were randomly divided into three groups, a sham group, a group subjected to 2h torsion followed by 24h detorsion and a group subjected to T/D and injected i.p. with 20mgkg-1 PD 30min before detorsion. Unilateral orchiectomy was performed after 24h of reperfusion. Half the testes were prepared for histological examination by haematoxylin-eosin staining and the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) technique...
April 26, 2017: Reproduction, Fertility, and Development
https://www.readbyqxmd.com/read/28372729/development-of-a-thermostabilised-triplex-lamp-assay-with-dry-reagent-four-target-lateral-flow-dipstick-for-detection-of-entamoeba-histolytica-and-non-pathogenic-entamoeba-spp
#6
Phiaw Chong Foo, Yean Yean Chan, Maizan Mohamed, Weng Kin Wong, A B Nurul Najian, Boon Huat Lim
This study highlighted the development of a four target nitrocellulose-based nucleic acid lateral flow immunoassay biosensor in a dry-reagent strip format for interpretation of double-labelled double-stranded amplicons from thermostabilised triplex loop-mediated isothermal amplification assay. The DNA biosensor contained two test lines which captured biotin and texas red labelled amplicons; a LAMP internal amplification control line that captured digoxigenin labelled amplicon; and a chromatography control line that validated the functionality of the conjugated gold nanoparticles and membrane...
May 8, 2017: Analytica Chimica Acta
https://www.readbyqxmd.com/read/28363012/fluorescence-image-analyzer-flima-software-for-quantitative-analysis-of-fluorescence-in-situ-hybridization
#7
H C M Silva, M M C Martins-Júnior, L B Ribeiro, D A Matoso
The Fluorescence Image Analyzer (FLIMA) software was developed for the quantitative analysis of images generated by fluorescence in situ hybridization (FISH). Currently, the images of FISH are examined without a coefficient that enables a comparison between them. Through GD Graphics Library, the FLIMA software calculates the amount of pixels on image and recognizes each present color. The coefficient generated by the algorithm shows the percentage of marks (probes) hybridized on the chromosomes. This software can be used for any type of image generated by a fluorescence microscope and is able to quantify digoxigenin probes exhibiting a red color, biotin probes exhibiting a green color, and double-FISH probes (digoxigenin and biotin used together), where the white color is displayed...
March 30, 2017: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/28349827/staurosporine-suppresses-survival-of-hepg2-cancer-cells-through-omi-htra2-mediated-inhibition-of-pi3k-akt-signaling-pathway
#8
Youming Ding, Bin Wang, Xiaoyan Chen, Yu Zhou, Jianhui Ge
Staurosporine, which is an inhibitor of a broad spectrum of protein kinases, has shown cytotoxicity on several human cancer cells. However, the underlying mechanism is not well understood. In this study, we examined whether and how this compound has an inhibitory action on phosphatidylinositol 3-kinase (PI3K)/Akt pathway in vitro using HepG2 human hepatocellular carcinoma cell line. Cell viability and apoptosis were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay, respectively...
March 2017: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
https://www.readbyqxmd.com/read/28334717/a-tandemly-arranged-pattern-of-two-5s-rdna-arrays-in-amolops-mantzorum-anura-ranidae
#9
Ting Liu, Menghuan Song, Yun Xia, Xiaomao Zeng
In an attempt to extend the knowledge of the 5S rDNA organization in anurans, the 5S rDNA sequences of Amolops mantzorum were isolated, characterized, and mapped by FISH. Two forms of 5S rDNA, type I (209 bp) and type II (about 870 bp), were found in specimens investigated from various populations. Both of them contained a 118-bp coding sequence, readily differentiated by their non-transcribed spacer (NTS) sizes and compositions. Four probes (the 5S rDNA coding sequences, the type I NTS, the type II NTS, and the entire type II 5S rDNA sequences) were respectively labeled with TAMRA or digoxigenin to hybridize with mitotic chromosomes for samples of all localities...
2017: Cytogenetic and Genome Research
https://www.readbyqxmd.com/read/28261779/does-protocatechuic-acid-a-natural-antioxidant-reduce-renal-ischemia-reperfusion-injury-in-rats
#10
Melih Yüksel, Murat Yıldar, Murat Başbuğ, Faruk Çavdar, Öztekin Çıkman, Hasan Akşit, Figen Aslan, Dilek Akşit
BACKGROUND: Protocatechuic acid (PCA), which has antioxidant property, is a simple phenolic compound commonly found in many plants, vegetables, and fruits, notably in green tea and almonds. Present study was an investigation of the effects of PCA on rat kidney with ischemia/reperfusion (IR) injury. METHODS: Sprague-Dawley rats were randomly divided into 4 groups: (1) Sham, (2) Renal IR, (3) Renal IR+Vehicle, and (4) Renal IR+PCA. Renal reperfusion injury was induced by clamping renal pedicle for 45 minutes after right nephrectomy was performed, followed by reperfusion for 3 hours...
January 2017: Ulusal Travma Ve Acil Cerrahi Dergisi, Turkish Journal of Trauma & Emergency Surgery: TJTES
https://www.readbyqxmd.com/read/28257848/molecular-characterization-of-pacific-white-shrimp-litopenaeus-vannamei-sodium-bicarbonate-cotransporter-nbc-and-its-role-in-response-to-ph-stress
#11
Yi-Ming Cai, Ting Chen, Chun-Hua Ren, Wen Huang, Xiao Jiang, Yan Gao, Da Huo, Chao-Qun Hu
The sodium bicarbonate cotransporter (NBC) is an integral membrane ion transporter that can transport HCO3(-) (or a related species, such as CO3(2-)) across the plasma membrane. Previous researches revealed that NBC might play an important role in the regulation of intracellular pH in vertebrates. In the present study, an NBC cDNA was identified from Pacific white shrimp (Litopenaeus vannamei) and designated as Lv-NBC. The full-length Lv-NBC cDNA is 4479 bp in size, containing a 5'-untranslated region (UTR) of 59 bp, a 3'-UTR of 835 bp and an open reading frame (ORF) of 3585 bp that encodes a protein of 1194 amino acids with a deduced molecular weight of 134...
May 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28245157/developing-stage-dependent-retinal-toxicity-induced-by-l-glutamate-in-neonatal-rats
#12
Hikaru Mitori, Takeshi Izawa, Mitsuru Kuwamura, Masahiro Matsumoto, Jyoji Yamate
The neurotransmitter glutamate causes excitotoxicity in the human retina. In neonatal rats, the degree of glutamate-induced retinal damage depends on age at administration. To elucidate the sensitivity to glutamate on various developing stage of retina, we investigated glutamate-induced retinal damage and glutamate target cells on each postnatal day (PND). Newborn rats received a single subcutaneous administration of l-glutamate on PNDs 1 to 14. Retinal cell apoptosis characterized as pyknotic and terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick end labeling-positive nuclei was analyzed at 6 hr after treatment, and sequential morphological features of retina were evaluated on PND 21...
December 2016: Toxicologic Pathology
https://www.readbyqxmd.com/read/28193316/polydatin-improves-the-developmental-competence-of-bovine-embryos-in-vitro-via-induction-of-sirtuin-1-sirt1
#13
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
#14
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...
May 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28133587/non-radioactive-trf-assay-modifications-to-improve-telomeric-dna-detection-efficiency-in-plants
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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