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Validation of Reference Genes for mRNA Quantification in Adjuvant Arthritis

Open Journal of Rheumatology and Autoimmune Diseases 2012年第3期2卷 64-72页

作者： Muhammad Ayaz Alam Qureshi Aisha Siddiqah Ahmed Jian Li andré stark Per Eriksson Mahmood Ahmed Department of Clinical Neurosciences Karolinska Institutet Stockholm Sweden Department of Clinical Sciences Danderyd Hospital Karolinska Institutet Stockholm Sweden Department of Medicine Karolinska University Hospital Karolinska Institutet Stockholm Sweden Department of Molecular Medicine and Surgery Karolinska University Hospital Karolinska Institutet Stockholm Sweden Department of Neurobiology Care Sciences and Society Karolinska Institutet Stockholm Sweden

Real time quantitative PCR (RT-qPCR) requires a method to normalize the expression of target genes against an en- dogenous reference gene. It is known that commonly used housekeeping genes (HKGs) vary tremendously in inflam- matory conditions;however information about the stability and expression of HKGs in chronic inflammatory joint dis- ease such as rheumatoid arthritis (RA) is scarce. The expressional stability of 10 commonly used HKGs was analyzed in the neuronal (spinal cord, dorsal root ganglia) and in the musculoskeletal tissues (tendon, muscle, epiphysis, capsule, periosteum and ankle joint) using RT-qPCR in the rat model of RA. In individual tissues, suitable HKGs were selected by | △Ct| (│Ct control-Ct arthritis│) and further analyzed by using software programs;geNorm and normfinder. We found hypoxanthine-guanine phosphoribosyl tranferase (HPRT) as the most stable gene except ankle joint while glyce-raldehyde-3-phosphate dehydrogenase (GAPDH) was found as the least stable gene in musculoskeletal tissues. In in-flamed ankle joint where no reference gene was found to be stably expressed, an inflammatory cell marker CD3 was used to normalize peptidylprolyl isomerase B (PPIB), the most homogenous HKG identified among the 10 HKGs. The normalized PPIB was then used to analyze the gene expression of neurokinin 1 (NK1), receptor of substance P, a potent pro-inflammatory mediator. We observed a 3.5 fold increase (p = 0.009) in NK1 expression in inflamed ankle joint compared to control. Our results indicate that reference genes stability should be evaluated before using them as refer- ence during inflammatory conditions. In tissues with intense inflammatory cell infiltration, an inflammatory cell marker should be used to normalize the selected reference gene to avoid erroneous results.

关键词： Quantitative PCR Adjuvant Arthritis Housekeeping Gene

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Variability of platelet aggregation in patients with clopidogrel treatment and hip fracture: A retrospective case-control study on 112 patients

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World Journal of Orthopedics 2015年第5期6卷 439-445页

作者： Anna Clareus Inga Fredriksson Hkan Wallén Max Gordon andré stark Olof Skldenberg Division of Orthopaedics Department of Clinical Sciences at Danderyd Hospital Karolinska Institutet Division of Anesthesiology Department of Clinical Sciences at Danderyd Hospital Karolinska Institutet Division of Cardiology Department of Clinical Sciences at Danderyd Hospital Karolinska Institute

AIM: To identify the rate of non-responders to clopidogrel treatment in hip fracture patients and study how non-responders differ from ***: In a retrospective case-control study we included 28 cases of acute proximal femoral fracture with clopidogrel treatment 2011 to 2013. Eighty-four controls from the same time period were included. Data collected included response to clopidogrel measured with multiple electrode aggregometry(MEA), intraoperative bleeding, erythrocyte transfusion, time to surgery and the incidence of adverse events up to 3 mo after surgery. RESULTS: Eight(29%) of the 28 cases were nonresponders. The median intraoperative bleeding was 300 mL(range, 0-1500), and was lower for non-responders(50 m L) but did not reach statistical significance. Erythrocyte transfusions did not differ between responders, non-responders and controls. Forty-five(40%) of 112 patients had adverse events postoperatively but the rate did not differ between patients with and without clopidogrel ***: Almost one-third of patients withclopidogrel treatment and an acute proximal femoral fracture are non-responders to antiplatelet therapy and can be operated without delay.

关键词： Proximal femoral fracture Clopidogrel Variability Bleeding Adverse events

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Validation of Reference Genes for mRNA Quantification in Adjuvant Arthritis

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Open Journal of Rheumatology and Autoimmune Diseases（风湿病与自身免疫疾病期刊） 2012年第3期

作者： Muhammad Ayaz Alam Qureshi Aisha Siddiqah Ahmed Jian Li andré stark Per Eriksson Mahmood Ahmed

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Expression of the Genes OsNRT1.1, OsNRT2.1, OsNRT2.2, and Kinetics of Nitrate Uptake in Genetically Contrasting Rice Varieties

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American Journal of Plant Sciences 2015年第2期6卷 306-314页

作者： Osmário J. L. Araújo Milena S. Pinto Marcus V. L. Sperandio Leandro A. Santos Elvia M. L. M. stark Manlio S. Fernandes andré Marques dos Santos Sonia Regina de Souza Plant Nutrition Laboratory Department of Soil Universidade Federal Rural do Rio de Janeiro Seropédica Brazil Plant Biochemistry Laboratory Department of Chemistry Universidade Federal Rural do Rio de Janeiro Seropédica Brazil

Four genetically contrasting rice varieties (IAC-47, Bico Ganga, Arroz de Revenda and Manteiga) according to Random Amplified Polymorphic DNA (RAPD) analysis were assessed regarding expression of the genes OsNRT1.1, OsNRT2.1 and OsNRT2.2 and the nitrate uptake kinetics parameters (Km and Vmax). Up to 250-fold increases in the induction of gene expression after nitrate resupply were observed for the high-affinity transporter (OsNRT2.1 and OsNRT2.2). However, no significant variations in Vmax among the varieties were obtained. The lower value of Km of the IAC-47 cultivar in relation to the Arroz de Revenda variety suggests a greater role of high-affinity transporter genes. These results indicate that closer attention should be paid to the expression levels of these genes in selecting varieties aiming to enhance nitrogen uptake efficiency.

关键词： RAPD Nitrogen Nitrogen Use Efficiency Oryza sativa L.

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