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https://www.readbyqxmd.com/read/27210246/selection-of-reference-genes-for-gene-expression-studies-related-to-lung-injury-in-a-preterm-lamb-model
#1
Prue M Pereira-Fantini, Anushi E Rajapaksa, Regina Oakley, David G Tingay
Preterm newborns often require invasive support, however even brief periods of supported ventilation applied inappropriately to the lung can cause injury. Real-time quantitative reverse transcriptase-PCR (qPCR) has been extensively employed in studies of ventilation-induced lung injury with the reference gene 18S ribosomal RNA (18S RNA) most commonly employed as the internal control reference gene. Whilst the results of these studies depend on the stability of the reference gene employed, the use of 18S RNA has not been validated...
2016: Scientific Reports
https://www.readbyqxmd.com/read/26968891/comprehensive-selection-of-reference-genes-for-expression-studies-in-meniscus-injury-using-quantitative-real-time-pcr
#2
Mariana Ferreira Leal, Gustavo Gonçalves Arliani, Diego Costa Astur, Carlos Eduardo Franciozi, Pedro Debieux, Carlos Vicente Andreoli, Marília Cardoso Smith, Alberto de Castro Pochini, Benno Ejnisman, Moises Cohen
The meniscus plays critical roles in the knee function. Meniscal tears can lead to knee osteoarthritis. Gene expression analysis may be a useful tool for understanding meniscus tears, and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) has become an effective method for such studies. However, this technique requires the use of suitable reference genes for data normalization. We evaluated the suitability of six reference genes (18S, ACTB, B2M, GAPDH, HPRT1 and TBP) using meniscus samples of (1) 19 patients with isolated meniscal tears, (2) 20 patients with meniscal tears and combined anterior cruciate ligament injury (ACL), and (3) 11 controls without meniscal tears...
June 10, 2016: Gene
https://www.readbyqxmd.com/read/26904058/selection-of-reference-genes-for-normalization-of-microrna-expression-by-rt-qpcr-in-sugarcane-buds-under-cold-stress
#3
Yuting Yang, Xu Zhang, Yun Chen, Jinlong Guo, Hui Ling, Shiwu Gao, Yachun Su, Youxiong Que, Liping Xu
Sugarcane, accounting for 80% of world's sugar, originates in the tropics but is cultivated mainly in the subtropics. Therefore, chilling injury frequently occurs and results in serious losses. Recent studies in various plant species have established microRNAs as key elements in the post-transcriptional regulation of response to biotic and abiotic stresses including cold stress. Though, its accuracy is largely influenced by the use of reference gene for normalization, quantitative PCR is undoubtedly a popular method used for identification of microRNAs...
2016: Frontiers in Plant Science
https://www.readbyqxmd.com/read/26192306/identification-of-suitable-reference-genes-for-investigating-gene-expression-in-anterior-cruciate-ligament-injury-by-using-reverse-transcription-quantitative-pcr
#4
Mariana Ferreira Leal, Diego Costa Astur, Pedro Debieux, Gustavo Gonçalves Arliani, Carlos Eduardo Silveira Franciozi, Leonor Casilla Loyola, Carlos Vicente Andreoli, Marília Cardoso Smith, Alberto de Castro Pochini, Benno Ejnisman, Moises Cohen
The anterior cruciate ligament (ACL) is one of the most frequently injured structures during high-impact sporting activities. Gene expression analysis may be a useful tool for understanding ACL tears and healing failure. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) has emerged as an effective method for such studies. However, this technique requires the use of suitable reference genes for data normalization. Here, we evaluated the suitability of six reference genes (18S, ACTB, B2M, GAPDH, HPRT1, and TBP) by using ACL samples of 39 individuals with ACL tears (20 with isolated ACL tears and 19 with ACL tear and combined meniscal injury) and of 13 controls...
2015: PloS One
https://www.readbyqxmd.com/read/26105877/pp018-does-altitude-affect-the-placental-renin-angiotensin-system-ras-in-pre-eclampsia-pe
#5
Lesia Kurlak, Hiten Mistry, Tereza Cindrova-Davies, Martha Tissot van Patot, Graham Burton, Fiona Broughton Pipkin
INTRODUCTION: We have previously shown the activation of placental RAS in high altitude normotensive (NT) pregnancies [1], presumably related to hypoxia during placental development. PE incidence is increased at high altitude; hypoxia-reoxygenation injury contributes to the elevated oxidative stress seen in this condition. Binding of Angiotensin II to its type I receptor (AT1R) increases generation of reactive oxygen species and we have shown an increase in AT1R in PE [2]. Therefore we investigated whether the RAS is further augmented in PE at high altitudes...
April 2013: Pregnancy Hypertension
https://www.readbyqxmd.com/read/25936423/selection-of-suitable-reference-genes-for-reverse-transcription-quantitative-polymerase-chain-reaction-analysis-of-neuronal-cells-differentiated-from-bone-mesenchymal-stem-cells
#6
Yu-Xi He, Yan Zhang, Qiwei Yang, Chenguang Wang, Guanfang Su
Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is a technique widely used for the quantification of mRNA transcription, It is well recognized that the reference genes used in RT-qPCR require appropriate validation to ensure that gene expression is unaffected by experimental conditions. The differentiation of bone mesenchymal stem cells (BMSCs) into neurons is important in the treatment of nerve injury. In gene expression analysis of the differentiation of BMSCs into neuronal cells by, the commonly used reference genes for RNA analysis are often selected without any preliminary evaluation of their suitability...
August 2015: Molecular Medicine Reports
https://www.readbyqxmd.com/read/25768100/identification-of-suitable-reference-genes-for-gene-expression-studies-in-tendons-from-patients-with-rotator-cuff-tear
#7
Mariana Ferreira Leal, Paulo Santoro Belangero, Eduardo Antônio Figueiredo, Carina Cohen, Leonor Casilla Loyola, Carlos Vicente Andreoli, Marília Cardoso Smith, Alberto de Castro Pochini, Benno Ejnisman, Moises Cohen
Rotator cuff tear is one of the most common causes of shoulder dysfunction. Gene expression analysis may be a useful tool for understanding tendon tears and the failure of cuff healing, and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) has become an effective method for such studies. However, this technique requires the use of suitable reference genes for data normalization. Here, we evaluate the suitability of six reference genes (18S, ACTB, B2M, GAPDH, HPRT1 and TBP) using samples from the rotator cuff tendons of 28 individuals with tendon tears (3 tendons regions) and 8 controls (2 tendon regions); for the tear patients, we evaluated ruptured and non-ruptured tendon samples...
2015: PloS One
https://www.readbyqxmd.com/read/25187167/validation-of-four-reference-genes-for-quantitative-mrna-expression-studies-in-a-rat-model-of-inflammatory-injury
#8
Roxanne Y Walder, Anne-Sophie Wattiez, Stephanie R White, Blanca Marquez de Prado, Marta V Hamity, Donna L Hammond
BACKGROUND: Real-time quantitative PCR (qPCR) is a technique frequently used to measure changes in mRNA expression. To ensure validity of experimental findings, it is important to normalize the qPCR data to reference genes that are stable and unaffected by the experimental treatment to correct for variability among samples. Unlike in some models of neuropathic pain, reference genes for models of inflammatory injury have not been validated. This study examined four candidate reference genes in an effort to identify and validate optimal genes for normalization of transcriptional changes occurring in the dorsal horn of the spinal cord and the rostral ventromedial medulla (RVM) following intraplantar injection of complete Freund's adjuvant (CFA)...
2014: Molecular Pain
https://www.readbyqxmd.com/read/25144298/identification-and-validation-of-suitable-housekeeping-genes-for-normalizing-quantitative-real-time-pcr-assays-in-injured-peripheral-nerves
#9
Giovanna Gambarotta, Giulia Ronchi, Olivier Friard, Pantaleo Galletta, Isabelle Perroteau, Stefano Geuna
Injury to the peripheral nerve induces dramatic changes in terms of cellular composition that are reflected on RNA quality and quantity, making messenger RNA expression analysis very complex. Several commonly used housekeeping genes are regulated following peripheral nerve injury and are thus not suitable for quantitative real-time PCR normalization; moreover, the presence of pseudogenes in some of them impairs their use. To deal with this problem, we have developed a new method to identify new stable housekeeping genes based on publicly available microarray data on normal and injured nerves...
2014: PloS One
https://www.readbyqxmd.com/read/25122470/identification-of-suitable-reference-genes-for-gene-expression-studies-of-shoulder-instability
#10
Mariana Ferreira Leal, Paulo Santoro Belangero, Carina Cohen, Eduardo Antônio Figueiredo, Leonor Casilla Loyola, Alberto Castro Pochini, Marília Cardoso Smith, Carlos Vicente Andreoli, Sintia Iole Belangero, Benno Ejnisman, Moises Cohen
Shoulder instability is a common shoulder injury, and patients present with plastic deformation of the glenohumeral capsule. Gene expression analysis may be a useful tool for increasing the general understanding of capsule deformation, and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) has become an effective method for such studies. Although RT-qPCR is highly sensitive and specific, it requires the use of suitable reference genes for data normalization to guarantee meaningful and reproducible results...
2014: PloS One
https://www.readbyqxmd.com/read/23983612/exploring-valid-reference-genes-for-quantitative-real-time-pcr-analysis-in-plutella-xylostella-lepidoptera-plutellidae
#11
Wei Fu, Wen Xie, Zhuo Zhang, Shaoli Wang, Qingjun Wu, Yong Liu, Xiaomao Zhou, Xuguo Zhou, Youjun Zhang
Quantitative real-time PCR (qRT-PCR), a primary tool in gene expression analysis, requires an appropriate normalization strategy to control for variation among samples. The best option is to compare the mRNA level of a target gene with that of reference gene(s) whose expression level is stable across various experimental conditions. In this study, expression profiles of eight candidate reference genes from the diamondback moth, Plutella xylostella, were evaluated under diverse experimental conditions. RefFinder, a web-based analysis tool, integrates four major computational programs including geNorm, Normfinder, BestKeeper, and the comparative ΔCt method to comprehensively rank the tested candidate genes...
2013: International Journal of Biological Sciences
https://www.readbyqxmd.com/read/23841944/reverse-transcription-quantitative-real-time-polymerase-chain-reaction-reference-genes-in-the-spared-nerve-injury-model-of-neuropathic-pain-validation-and-literature-search
#12
Nicolas Piller, Isabelle Decosterd, Marc R Suter
BACKGROUND: The reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is a widely used, highly sensitive laboratory technique to rapidly and easily detect, identify and quantify gene expression. Reliable RT-qPCR data necessitates accurate normalization with validated control genes (reference genes) whose expression is constant in all studied conditions. This stability has to be demonstrated.We performed a literature search for studies using quantitative or semi-quantitative PCR in the rat spared nerve injury (SNI) model of neuropathic pain to verify whether any reference genes had previously been validated...
2013: BMC Research Notes
https://www.readbyqxmd.com/read/23702882/molecular-pathology-of-brain-edema-after-severe-burns-in-forensic-autopsy-cases-with-special-regard-to-the-importance-of-reference-gene-selection
#13
COMPARATIVE STUDY
Qi Wang, Takaki Ishikawa, Tomomi Michiue, Bao-Li Zhu, Da-Wei Guan, Hitoshi Maeda
Brain edema is believed to be linked to high mortality incidence after severe burns. The present study investigated the molecular pathology of brain damage and responses involving brain edema in forensic autopsy cases of fire fatality (n = 55) compared with sudden cardiac death (n = 11), mechanical asphyxia (n = 13), and non-brain injury cases (n = 22). Postmortem mRNA and immunohistochemical expressions of aquaporins (AQPs), claudin5 (CLDN5), and matrix metalloproteinases (MMPs) were examined...
September 2013: International Journal of Legal Medicine
https://www.readbyqxmd.com/read/23558424/identification-of-endogenous-reference-genes-for-rt-qpcr-analysis-of-plasma-micrornas-levels-in-rats-with-acetaminophen-induced-hepatotoxicity
#14
Yan Wang, Naping Tang, Taotao Hui, Shuyan Wang, Xiancheng Zeng, Hua Li, Jing Ma
Circulating microRNA (miRNA) expression profiles have been reported to be promising biomarkers for drug-induced liver injury in preclinical and clinical practice. Proper normalization is critical for accurate miRNAs expression analysis. Herein, using SYBR green quantitative real-time PCR (RT-qPCR), we evaluated the expression stability of six candidate reference genes including two commonly used small RNAs (U6, 5S) and four miRNAs (let-7a, miR-92a, miR-103 and miR-16) in plasma of rats with acetaminophen-induced hepatotoxicity...
November 2013: Journal of Applied Toxicology: JAT
https://www.readbyqxmd.com/read/23203068/selection-of-suitable-reference-genes-for-normalization-of-quantitative-real-time-pcr-in-cartilage-tissue-injury-and-repair-in-rabbits
#15
Xiao-Xiang Peng, Rong-Lan Zhao, Wei Song, Hai-Rong Chu, Meng Li, Shu-Ya Song, Guang-Zhou Li, Dong-Chun Liang
When studying the altered expression of genes associated with cartilage regeneration by quantitative real-time RT-PCR (RT-qPCR), reference genes with highly stable expression during different stages of chondrocyte developmental are necessary to normalize gene expression accurately. Until now, no reports evaluating expression changes of commonly used reference genes in rabbit articular cartilage have been published. In this study, defects were made in rabbit articular cartilage, with or without insulin-like growth factor 1 (IGF-1) treatment, to create different chondrocyte living environments...
2012: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/23146128/identification-of-stable-normalization-genes-for-quantitative-real-time-pcr-in-porcine-articular-cartilage
#16
Ryan S McCulloch, Melissa S Ashwell, Audrey T O'Nan, Peter L Mente
UNLABELLED: HASH(0x3b54330) BACKGROUND: Expression levels for genes of interest must be normalized with an appropriate reference, or housekeeping gene, to make accurate comparisons of quantitative real-time PCR results. The purpose of this study was to identify the most stable housekeeping genes in porcine articular cartilage subjected to a mechanical injury from a panel of 10 candidate genes. RESULTS: Ten candidate housekeeping genes were evaluated in three different treatment groups of mechanically impacted porcine articular cartilage...
2012: Journal of Animal Science and Biotechnology
https://www.readbyqxmd.com/read/23010907/stability-of-endogenous-reference-genes-in-postmortem-human-brains-for-normalization-of-quantitative-real-time-pcr-data-comprehensive-evaluation-using-genorm-normfinder-and-bestkeeper
#17
Qi Wang, Takaki Ishikawa, Tomomi Michiue, Bao-Li Zhu, Da-Wei Guan, Hitoshi Maeda
In forensic molecular pathology, quantitative real-time polymerase chain reaction (RT-qPCR) provides a rapid and sensitive method to investigate functional changes in the death process. Accurate and reliable relative RT-qPCR requires ideal amplification efficiencies of target and reference genes. However, the amplification efficiency, changing during PCR, may be overestimated by the traditional standard curve method. No single gene meets the criteria of an ideal endogenous reference. Therefore, it is necessary to select suitable reference genes for specific requirements...
November 2012: International Journal of Legal Medicine
https://www.readbyqxmd.com/read/22563491/reference-genes-for-real-time-pcr-quantification-of-micrornas-and-messenger-rnas-in-rat-models-of-hepatotoxicity
#18
María N Lardizábal, Ana L Nocito, Stella M Daniele, Leonardo A Ornella, Javier F Palatnik, Luis M Veggi
Hepatotoxicity is associated with major changes in liver gene expression induced by xenobiotic exposure. Understanding the underlying mechanisms is critical for its clinical diagnosis and treatment. MicroRNAs are key regulators of gene expression that control mRNA stability and translation, during normal development and pathology. The canonical technique to measure gene transcript levels is Real-Time qPCR, which has been successfully modified to determine the levels of microRNAs as well. However, in order to obtain accurate data in a multi-step method like RT-qPCR, the normalization with endogenous, stably expressed reference genes is mandatory...
2012: PloS One
https://www.readbyqxmd.com/read/22377061/selection-of-reference-genes-in-different-myocardial-regions-of-an-in-vivo-ischemia-reperfusion-rat-model-for-normalization-of-antioxidant-gene-expression
#19
Nicoletta Vesentini, Cristina Barsanti, Alessandro Martino, Claudia Kusmic, Andrea Ripoli, AnnaMaria Rossi, Antonio L'Abbate
BACKGROUND: Changes in cardiac gene expression due to myocardial injury are usually assessed in whole heart tissue. However, as the heart is a heterogeneous system, spatial and temporal heterogeneity is expected in gene expression. RESULTS: In an ischemia/reperfusion (I/R) rat model we evaluated gene expression of mitochondrial and cytoplasmatic superoxide dismutase (MnSod, Cu-ZnSod) and thioredoxin reductase (trxr1) upon short (4 h) and long (72 h) reperfusion times in the right ventricle (RV), and in the ischemic/reperfused (IRR) and the remote region (RR) of the left ventricle...
February 29, 2012: BMC Research Notes
https://www.readbyqxmd.com/read/21863315/quantification-of-gene-expression-after-painful-nerve-injury-validation-of-optimal-reference-genes
#20
Madhavi Latha Yadav Bangaru, Frank Park, Andy Hudmon, J Bruce McCallum, Quinn H Hogan
Stably expressed housekeeping genes (HKGs) are necessary for standardization of transcript measurement by quantitative real-time polymerase chain reaction (qRT-PCR). Peripheral nerve injury disrupts expression of numerous genes in sensory neurons, but the stability of conventional HKGs has not been tested in this context. We examined the stability of candidate HKGs during nerve injury, including the commonly used 18S ribosomal RNA, β-tubulin I and β-tubulin III, actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and hypoxanthine phosphoribosyltransferase 1 (HPRT1), and mitogen-activated protein kinase 6 (MAPK6)...
March 2012: Journal of Molecular Neuroscience: MN
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