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Biological Procedures Online

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https://www.readbyqxmd.com/read/30479579/anti-inflammatory-activity-of-tocotrienols-in-age-related-pathologies-a-saspected-involvement-of-cellular-senescence
#1
REVIEW
Marco Malavolta, Elisa Pierpaoli, Robertina Giacconi, Andrea Basso, Maurizio Cardelli, Francesco Piacenza, Mauro Provinciali
Tocotrienols (T3) have been shown to represent a very important part of the vitamin E family since they have opened new opportunities to prevent or treat a multitude of age-related chronic diseases. The beneficial effects of T3 include the amelioration of lipid profile, the promotion of Nrf2 mediated cytoprotective activity and the suppression of inflammation. All these effects may be the consequence of the ability of T3 to target multiple pathways. We here propose that these effects may be the result of a single target of T3, namely senescent cells...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/30410426/crispr-cas9-knockout-strategies-to-ablate-ccat1-lncrna-gene-in-cancer-cells
#2
Khadijeh Zare, Milad Shademan, Mohammad M Ghahramani Seno, Hesam Dehghani
Background: With the increasing discovery of long noncoding RNAs (lncRNAs), the application of functional techniques that could have very specific, efficient, and robust effects and readouts is necessary. Here, we have applied and analyzed three gene knockout (KO) strategies to ablate the CCAT1 gene in different colorectal adenocarcinoma cell lines. We refer to these strategies as "CRISPR excision", "CRISPR HDR", and "CRISPR du-HITI". Results: In order to obstruct the transcription of lncRNA or to alter its structure, in these strategies either a significant segment of the gene is removed, or a transcription termination signal is inserted in the target gene...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/30356830/three-dimensional-quantification-of-spheroid-degradation-dependent-invasion-and-invadopodia-formation
#3
Cameron Goertzen, Denise Eymael, Marco Magalhaes
Invadopodia are actin-rich, proteolytic structures that enable cancer cell to invade into the surrounding tissues. Several in vitro invasion assays have been used in the literature ranging from directional quantitative assays to complex three-dimensional (3D) analyses. One of the main limitations of these assays is the lack of quantifiable degradation-dependent invasion in a three-dimensional (3D) environment that mimics the tumor microenvironment. In this article, we describe a new invasion and degradation assay based on the currently available tumor spheroid model that allows long-term high-resolution imaging of the tumor, precise quantification, and visualization of matrix degradation and multichannel immunocytochemistry...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/30337841/modified-sureselect-qxt-target-enrichment-protocol-for-illumina-multiplexed-sequencing-of-ffpe-samples
#4
J M Rosa-Rosa, T Caniego-Casas, S Leskela, G Muñoz, F Del Castillo, P Garrido, J Palacios
Background: Personalised medicine is nowadays a major objective in oncology. Molecular characterization of tumours through NGS offers the possibility to find possible therapeutic targets in a time- and cost-effective way. However, the low quality and complexity of FFPE DNA samples bring a series of disadvantages for massive parallel sequencing techniques compared to high-quality DNA samples (from blood cells, cell cultures, etc.). Results: We performed several experiments to understand the behaviour of FFPE DNA samples during the construction of SureSelectQXT libraries...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/30275774/a-new-elisa-to-overcome-the-pitfalls-in-quantification-of-recombinant-human-monoclonal-anti-hbs-gc1102-by-commercial-immunoassays
#5
Yong Won Shin, Dong-Hyung Cho, Gi Won Song, Se-Ho Kim
Several methods for the quantification of human anti-HBs, an antibody to hepatitis B surface antigen (HBsAg), have been developed based on enzyme reaction, chemiluminescence, fluorescence, and radioactivity for application to human serum or plasma. Commercial anti-HBs immunoassay kits use a sandwich method in which a bridge is formed by the anti-HBs between a HBsAg immobilized solid matrix and the labeled HBsAg. However, this direct sandwich enzyme-linked immunosorbent assay (ELISA) is insufficient to accurately evaluate the activity of the human monoclonal anti-HBs, GC1102...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/30214383/microrna-regulation-in-estrogen-receptor-positive-breast-cancer-and-endocrine-therapy
#6
REVIEW
Erin W Howard, Xiaohe Yang
As de novo and acquired resistance to standard first line endocrine therapies is a growing clinical challenge for estrogen receptor-positive (ER+ ) breast cancer patients, understanding the mechanisms of resistance is critical to develop novel therapeutic strategies to prevent therapeutic resistance and improve patient outcomes. The widespread post-transcriptional regulatory role that microRNAs (miRNAs) can have on various oncogenic pathways has been well-documented. In particular, several miRNAs are reported to suppress ERα expression via direct binding with the 3' UTR of ESR1 mRNA, which can confer resistance to estrogen/ERα-targeted therapies...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/30093840/targeting-of-her3-with-functional-cooperative-mirnas-enhances-therapeutic-activity-in-her2-overexpressing-breast-cancer-cells
#7
Hui Lyu, Jingcao Huang, Zhimin He, Bolin Liu
Background: The HER3 receptor functions as a major cause of drug resistance in cancer treatment. It is believed that therapeutic targeting of HER3 is required to improve patient outcomes. It is not clear whether a novel strategy with two functional cooperative miRNAs would effectively inhibit erbB3 expression and potentiate the anti-proliferative/anti-survival effects of a HER2-targeted therapy (trastuzumab) and chemotherapy (paclitaxel) on HER2-overexpressing breast cancer cells. Results: Combination of miR-125a and miR-205, as compared to either miRNA alone, potently inhibited expression of HER3 in HER2-overexpressing breast cancer BT474 cells...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/30078998/real-time-vital-mineralization-detection-and-quantification-during-in-vitro-osteoblast-differentiation
#8
Anastassia Serguienko, Meng Yu Wang, Ola Myklebost
Background: In vitro studies of osteoblasts traditionally use Alizarin Red as a golden standard for the detection and quantification of mineralization, which is a marker of osteoblast differentiation. However, this method presents a number of drawbacks, including the need to fix cells, which prevents additional measurements. Years ago, Calcein Green was proposed as an alternative to Alizarin Red, with the advantage to be directly detectable in live cells. However, the protocol was still time-consuming, and it never managed to replace Alizarin Red...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29988526/optimization-of-immunofluorescent-detection-of-bone-marrow-disseminated-tumor-cells
#9
Haley D Axelrod, Kenneth J Pienta, Kenneth C Valkenburg
Background: Cancer metastasis is the primary cause of cancer-related deaths and remains incurable. Current clinical methods for predicting metastatic recurrence are not sensitive enough to detect individual cancer cells in the body; therefore, current efforts are directed toward liquid biopsy-based assays to capture circulating and disseminated tumor cells (CTCs and DTCs) in the blood and bone marrow, respectively. The most promising strategy is fluorescence-based immunostaining using cancer cell-specific markers...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29988477/icariin-a-potential-compound-for-the-recovery-of-tibial-dyschondroplasia-affected-chicken-via-up-regulating-bmp-2-expression
#10
Mujahid Iqbal, Hui Zhang, Khalid Mehmood, Aoyun Li, Xiong Jiang, Yaping Wang, Jialu Zhang, Muhammad Kashif Iqbal, Mujeeb Ur Rehman, Wangyuan Yao, Shijin Yang, Jiakui Li
Background: Tibial dyschondroplasia (TD) is a skeletal disease of fast growing chicken and other avian species. It is characterized by an avascular and non-mineralized growth plate, which leads to a deformed tibial bone and lameness. Unfortunately, this disease is not only responsible for causing huge economic losses but also raises animal welfare concerns. Icariin is a flavonoid, which is isolated from Epimedium pubescens herb, and it has been used to cure different diseases including bone fractures and osteoporosis...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29881330/method-for-improving-the-quality-of-genomic-dna-obtained-from-minute-quantities-of-tissue-and-blood-samples-using-chelex-100-resin
#11
Utkarsha A Singh, Mukta Kumari, Soumya Iyengar
Background: Although genomic DNA isolation using the Chelex 100 resin is rapid and inexpensive, the DNA obtained by this method has a low concentration in solution and contains suspended impurities. The presence of debris in the DNA solution may result in degradation of DNA on long term storage and inhibition of the polymerase chain reaction. In order to remove impurities and concentrate the DNA in solution, we have introduced modifications in the existing DNA isolation protocol using Chelex-100...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29881329/isolation-and-semi-quantitative-pcr-of-na-h-antiporter-sos1-and-nhx-genes-under-salinity-stress-in-kochia-scoparia
#12
Leila Fahmideh, Ziba Fooladvand
Background: Kochia scoparia is a dicotyledonous annual herb and belongs to the Amaranthaceae family. Genetic diversity and resistance to drought stress of this plant has made it widely scattered in different regions which contains highly genetic diversity and great potential as fodder and can grow on salty, drought affected areas. Since the soil salinity has become widely spread, environmental concern has sparked so many debates. An important limiting factor in agricultural production worldwide is the sensitivity of most of the crop to salinity caused by high concentration of salts soil...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29743821/blockade-of-itga2-induces-apoptosis-and-inhibits-cell-migration-in-gastric-cancer
#13
Yu-Chang Chuang, Hsin-Yi Wu, Yu-Ling Lin, Shey-Cherng Tzou, Cheng-Hsun Chuang, Ting-Yan Jian, Pin-Rong Chen, Yuan-Ching Chang, Chi-Hsin Lin, Tse-Hung Huang, Chao-Ching Wang, Yi-Lin Chan, Kuang-Wen Liao
Background: Gastric cancer is currently the fourth leading cause of cancer-related death worldwide. Gastric cancer is often diagnosed at advanced stages and the outcome of the treatment is often poor. Therefore, identifying new therapeutic targets for this cancer is urgently needed. Integrin alpha 2 (ITGA2) subunit and the beta 1 subunit form a heterodimer for a transmembrane receptor for extracellular matrix, is an important molecule involved in tumor cell proliferation, survival and migration...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29743820/use-of-rna-immunoprecipitation-method-for-determining-sinorhizobium-meliloti-rna-hfq-protein-associations-in-vivo
#14
Mengsheng Gao, Anne Benge, Julia M Mesa, Regina Javier, Feng-Xia Liu
Background: Soil bacterium Sinorhizobium meliloti ( S. meliloti ) forms an endosymbiotic partnership with Medicago truncatula ( M. truncatula ) roots which results in root nodules. The bacteria live within root nodules where they function to fix atmospheric N2 and supply the host plant with reduced nitrogen. The bacterial RNA-binding protein Hfq (Hfq) is an important regulator for the effectiveness of the nitrogen fixation. RNA immunoprecipitation (RIP) method is a powerful method for detecting the association of Hfq protein with specific RNA in cultured bacteria, yet a RIP method for bacteria living in root nodules remains to be described...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29618954/treatment-of-advanced-non-small-cell-lung-cancer-with-qi-nourishing-essence-replenishing-chinese-herbal-medicine-combined-with-chemotherapy
#15
Yabin Gong, Zhenye Xu, Changjuan Jin, Haibin Deng, Zhongqi Wang, Weidong Zhou, Ming Zhang, Xiaozhen Zhao, Lifang Wang
Background: To evaluate the effect of qi-nourishing essence-replenishing Chinese herbal medicine combined with chemotherapy in survival of advanced non-small-cell lung cancer(NSCLC) patients with essence and qi deficiency. Methods: A prospective multi-centered randomized controlled study was conducted, and 266 advanced NSCLC patients were enrolled. 126 patients in control group received Vinorelbine plus cisplatin(NP) chemotherapy combined with symptom-oriented Chinese herbs medication(without qi-nourishing essence-replenishing herbs);140 patients in experimental group received NP chemotherapy combined with qi-nourishing essence-replenishing Chinese herbal medication(Kangliu Zengxiao Decoction and modified Feiyanning Decoction, during and after chemotherapy respectively)...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29618953/dna-area-and-netosis-analysis-dana-a-high-throughput-method-to-quantify-neutrophil-extracellular-traps-in-fluorescent-microscope-images
#16
Ryan Rebernick, Lauren Fahmy, Christopher Glover, Mandar Bawadekar, Daeun Shim, Caitlyn L Holmes, Nicole Rademacher, Hemanth Potluri, Christie M Bartels, Miriam A Shelef
Background: Neutrophil extracellular traps (NETs), extracellular structures composed of decondensed chromatin and antimicrobial molecules, are released in a process called NETosis. NETs, which are part of normal host defense, have also been implicated in multiple human diseases. Unfortunately, methods for quantifying NETs have limitations which constrain the study of NETs in disease. Establishing optimal methods for NET quantification holds the potential to further elucidate the role of NETs in normal and pathologic processes...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29507535/high-sensitivity-and-high-resolution-in-situ-hybridization-of-coding-and-long-non-coding-rnas-in-vertebrate-ovaries-and-testes
#17
Natsumi Takei, Takuma Nakamura, Shohei Kawamura, Yuki Takada, Yui Satoh, Atsushi P Kimura, Tomoya Kotani
Background: Subcellular localization of coding and non-coding RNAs has emerged as major regulatory mechanisms of gene expression in various cell types and many organisms. However, techniques that enable detection of the subcellular distribution of these RNAs with high sensitivity and high resolution remain limited, particularly in vertebrate adult tissues and organs. In this study, we examined the expression and localization of mRNAs encoding Pou5f1/Oct4, Mos, Cyclin B1 and Deleted in Azoospermia-like (Dazl) in zebrafish and mouse ovaries by combining tyramide signal amplification (TSA)-based in situ hybridization with paraffin sections which can preserve cell morphology of tissues and organs at subcellular levels...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29507534/silhouette-scores-for-arbitrary-defined-groups-in-gene-expression-data-and-insights-into-differential-expression-results
#18
Shitao Zhao, Jianqiang Sun, Kentaro Shimizu, Koji Kadota
Background: Hierarchical Sample clustering (HSC) is widely performed to examine associations within expression data obtained from microarrays and RNA sequencing (RNA-seq). Researchers have investigated the HSC results with several possible criteria for grouping (e.g., sex, age, and disease types). However, the evaluation of arbitrary defined groups still counts in subjective visual inspection. Results: To objectively evaluate the degree of separation between groups of interest in the HSC dendrogram, we propose to use Silhouette scores...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29434528/antitumor-efficacy-of-human-monocyte-derived-dendritic-cells-comparing-effects-of-two-monocyte-isolation-methods
#19
Graça S Marques, Zélia Silva, Paula A Videira
Background: Dendritic cells (DCs), which can be used as anti-cancer vaccines, are generally obtained in vitro from isolated CD14+ monocytes (MoDCs). This generates high cell numbers and allows instructing DCs to guarantee effective antitumor responses. However, the impact of the monocyte isolation step in the antitumor effectiveness of the generated MoDCs is still unknown. Here, we compared the most used immunomagnetic technologies for monocyte isolation: magnetic activated cell sorting (MACS) from Miltenyi Biotec and EasySep from STEM CELL...
2018: Biological Procedures Online
https://www.readbyqxmd.com/read/29434527/visualisation-of-multiple-tight-junctional-complexes-in-human-airway-epithelial-cells
#20
Alysia G Buckley, Kevin Looi, Thomas Iosifidis, Kak-Ming Ling, Erika N Sutanto, Kelly M Martinovich, Elizabeth Kicic-Starcevich, Luke W Garratt, Nicole C Shaw, Francis J Lannigan, Alexander N Larcombe, Graeme Zosky, Darryl A Knight, Paul J Rigby, Anthony Kicic, Stephen M Stick
Background: Apically located tight junctions in airway epithelium perform a fundamental role in controlling macromolecule migration through paracellular spaces. Alterations in their expression may lead to disruptions in barrier integrity, which subsequently facilitates entry of potential bacterial and other pathogens into the host. Furthermore, there is emerging evidence that the barrier integrity of the airway in certain airway inflammatory diseases may be altered. However, there is little consensus on the way this is assessed and measured and the type of cells used to achieve this...
2018: Biological Procedures Online
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