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https://www.readbyqxmd.com/read/29140979/ginsenoside-rg3-inhibits-angiogenesis-in-a-rat-model-of-endometriosis-through-the-vegfr-2-mediated-pi3k-akt-mtor-signaling-pathway
#1
Yang Cao, Qing Ye, Mengfei Zhuang, Shuwu Xie, Ruihua Zhong, Jingang Cui, Jieyun Zhou, Yan Zhu, Tingting Zhang, Lin Cao
OBJECTIVE: This study aimed to investigate the link between the inhibitory effect of ginsenoside Rg3 on the ectopic endometrium growth and the VEGFR-2-mediated PI3K/Akt/mTOR signaling pathway, a mechanism known to inhibit angiogenesis and induce ectopic endometrial cell apoptosis. MATERIALS AND METHODS: A model of endometriosis was established by allotransplantation in rats. The rats were randomly divided into 5 groups: the ginsenoside Rg3 low-dose group (group A,5mg/kgBW/d of ginsenoside Rg3), the ginsenoside Rg3 high-dose group (group B, 10mg/kgBW/d of ginsenoside Rg3), the gestrinone group (group C, 0...
2017: PloS One
https://www.readbyqxmd.com/read/29069529/improved-free-energy-landscape-quantification-illustrated-with-a-computationally-designed-protein-ligand-interaction
#2
William J Van Patten, Robert Walder, Ayush Adhikari, Stephen R Okoniewski, Rashmi Ravichandran, Christine E Tinberg, David Baker, Thomas T Perkins
Quantifying the energy landscape underlying protein-ligand interactions leads to an enhanced understanding of molecular recognition. A powerful yet accessible single-molecule technique is atomic force microscopy (AFM)-based force spectroscopy, which generally yields the zero-force dissociation rate constant (koff) and the distance to the transition state (Δx‡). Here, we introduce an enhanced AFM assay and apply it to probe the computationally designed protein DIG10.3 binding to its target ligand, digoxigenin...
October 25, 2017: Chemphyschem: a European Journal of Chemical Physics and Physical Chemistry
https://www.readbyqxmd.com/read/29027177/spike-in-normalization-of-chip-data-using-dna-dig-antibody-complex
#3
Andrea B Eberle
Chromatin immunoprecipitation (ChIP) is a widely used method to determine the occupancy of specific proteins within the genome, helping to unravel the function and activity of specific genomic regions. In ChIP experiments, normalization of the obtained data by a suitable internal reference is crucial. However, particularly when comparing differently treated samples, such a reference is difficult to identify. Here, a simple method to improve the accuracy and reliability of ChIP experiments by the help of an external reference is described...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28944875/asymmetric-dimethylarginine-downregulates-sarco-endoplasmic-reticulum-calcium%C3%A2-atpase-3-and-induces-endoplasmic-reticulum-stress-in-human-umbilical-vein-endothelial-cells
#4
Weikang Guo, Zongli Diao, Wenhu Liu
Cardiovascular disease is the leading cause of mortality in patients with chronic kidney disease. Endothelial cell injury and apoptosis may promote atherosclerosis and cardiovascular disease. The present study investigated the potential mechanisms of asymmetric dimethylarginine (ADMA)‑induced apoptosis in human umbilical vein endothelial cells (HUVECs). It was demonstrated that ADMA decreased B‑cell lymphoma‑2 expression and increased cleaved‑caspase‑3 expression. Furthermore, terminal deoxynucleotidyl transferase (TdT)‑mediated‑digoxigenin‑11‑dUTP nick end labeling results indicated that ADMA induced apoptosis in HUVECs...
November 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28912690/ring-finger-protein-38-is-a-neuronal-protein-in-the-brain-of-nile-tilapia-oreochromis-niloticus
#5
Kai Lin Cham, Tomoko Soga, Ishwar S Parhar
Really interesting new gene (RING) finger protein is a type of zinc-binding motif found in a large family of functionally distinct proteins. RING finger proteins are involved in diverse cellular processes including apoptosis, DNA repair, cell cycle, signal transduction, tumour suppressor, vesicular transport, and peroxisomal biogenesis. RING finger protein 38 (RNF38) is a member of the family whose functions remain unknown. To gain insight into the putative effects of RNF38 in the central nervous system, we localised its expression...
2017: Frontiers in Neuroanatomy
https://www.readbyqxmd.com/read/28882513/enhanced-detection-of-rickettsia-species-in-ixodes-pacificus-using-highly-sensitive-fluorescence-in-situ-hybridization-coupled-with-tyramide-signal-amplification
#6
Ghazaleh Bagheri, Jeremy D Lehner, Jianmin Zhong
Ixodes pacificus is a host of many bacteria including Rickettsia species phylotypes G021 and G022. As part of the overall goal of understanding interactions of phylotypes with their tick host, this study focused on molecular detection of rickettsiae in ovarian and midgut tissue of I. pacificus by fluorescent in situ hybridization (FISH), PCR, and ultrastructural analysis. Of three embedding media (Technovit 8100, Unicryl, and paraffin) tested for generating thin sections, tissues embedded in paraffin resulted in the visualization of bacteria with low autofluorescence in FISH...
October 2017: Ticks and Tick-borne Diseases
https://www.readbyqxmd.com/read/28866013/contrasting-cellular-damage-after-blue-iris-and-femto-lasik-in-cat-cornea
#7
Kaitlin T Wozniak, Noah Elkins, Daniel R Brooks, Daniel E Savage, Scott MacRae, Jonathan D Ellis, Wayne H Knox, Krystel R Huxlin
Blue-intra-tissue refractive index shaping (Blue-IRIS) is a new approach to laser refractive correction of optical aberrations in the eye, which alters the refractive index of the cornea rather than changing its shape. Before it can be implemented in humans, it is critical to establish whether and to what extent, Blue-IRIS damages the cornea. Here, we contrasted the impact of -1.5 D cylinder refractive corrections inscribed using either Blue-IRIS or femtosecond laser in-situ keratomileusis (femto-LASIK) on corneal cell viability...
August 31, 2017: Experimental Eye Research
https://www.readbyqxmd.com/read/28810735/accurate-electrochemistry-analysis-of-circulating-methylated-dna-from-clinical-plasma-based-on-paired-end-tagging-and-amplifications
#8
Feng Chen, Xuyao Wang, Xiaowen Cao, Yongxi Zhao
Circulating methylated DNA has been a new kind of cancer biomarker, yet its small fraction of trace total DNA from clinical samples impairs the accurate analysis. Though fluorescence methods based on quantitative methylation specific PCR (qMSP) have been adopted routinely, yet alternative electrochemistry assay of such DNA from clinical samples remains a great challenge. Herein, we report accurate electrochemistry analysis of circulating methylated DNA from clinical plasma samples based on a paired-end tagging and amplifications strategy...
September 11, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28754459/first-description-in-europe-of-the-emergence-of-enterococcus-faecium-st117-carrying-both-vana-and-vanb-genes-isolated-in-greece
#9
C C Papagiannitsis, E Malli, Z Florou, M Medvecky, S Sarrou, J Hrabak, E Petinaki
OBJECTIVES: An Enterococcus faecium isolate (Efa-125) carrying both the vanA and vanB genes was recovered from a patient with bacteraemia treated in a Greek hospital. Since this is the first description in Europe of E. faecium carrying both vanA and vanB genes, the isolate was further studied. METHODS: Susceptibility to several antibiotics was determined using the VITEK(®)2 automated system. The isolate was typed by multilocus sequence typing (MLST). To define the genetic units of the vanA and vanB genes, the plasmid content of Efa-125 was analysed by pulsed-field gel electrophoresis (PFGE) of total DNA digested with S1 nuclease followed by hybridisation with digoxigenin-labelled vanA and vanB probes...
July 25, 2017: Journal of Global Antimicrobial Resistance
https://www.readbyqxmd.com/read/28748483/expression-of-microrna-146a-and-microrna-155-in-placental-villi-in-early-and-late-onset-preeclampsia
#10
N V Nizyaeva, G V Kulikova, M N Nagovitsyna, N E Kan, K N Prozorovskaya, A I Shchegolev, G T Sukhikh
We studied the expression of microRNA-146a and microRNA-155 in placental villi from 18 women (26-39 weeks of gestation) of reproductive age with early- or late-onset preeclampsia. The reference group consisted of women with physiological pregnancy and full-term gestation and with preterm birth after caesarian section on gestation week 26-31. MicroRNA-146a and microRNA-155 were detected by in situ hybridization with digoxigenin on paraffin sections. It was found that the expression of microRNA-146a in both syncytiotrophoblast of the intermediate villi and syncytial knots was lower at late-onset preeclampsia than at physiologic pregnancy of full-term period (p=0...
July 27, 2017: Bulletin of Experimental Biology and Medicine
https://www.readbyqxmd.com/read/28748477/chromogenic-in-situ-hybridization-methods-for-microrna-biomarker-monitoring-of-drug-safety-and-efficacy
#11
Barbara R Gould, Tina Damgaard, Boye Schnack Nielsen
Disease research and treatment development have turned to the impact and utility of microRNA. The dynamic and highly specific expression of these molecular regulators can be used to predict and monitor disease progression as well as therapeutic treatment efficacy and safety, thus aiding decisions in patient care. In situ hybridization (ISH) of biopsy material has become a routine clinical pathology procedure for monitoring gene structure, expression, and sample characterization. For ribonucleic acid (RNA), determining cell source and level of expression of these biomarkers gives insight into the cellular function and physiopathology...
2017: Methods in Molecular Biology
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
#12
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...
August 1, 2017: Bioorganic & Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/28608344/nonradioactive-detection-of-small-rnas-using-digoxigenin-labeled-probes
#13
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
#14
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...
October 2017: Archives of Oral Biology
https://www.readbyqxmd.com/read/28573617/nonradioactive-northern-blot-analysis-to-detect-ebola-virus-minigenomic-mrna
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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