Abstract:
BACKGROUND:Assessment of mRNA levels has the potential to predict renal outcome. The objective of this study was to optimize several steps in the protocol for obtaining cDNA from routine clinical kidney biopsy material. METHODS:RNA integrity was compared between different methods for extraction of RNA, synthesis of cDNA, and storage of renal tissue. Hereby, RNAlater, an RNA-preserving compound, was tested for implementation in a protocol for renal biopsies that combines routine histology and RNA expression studies. Gel electrophoresis and real-time polymerase chain reactions (PCR) were outcome parameters for assessment of RNA integrity. RESULTS:The Trizol method rendered higher RNA yields from fresh renal tissue than the NP40 method and RNeasy spin columns. RNA yields were not affected when renal tissue was stored at -70 degrees C for up to 2 months in phosphate-buffered saline (PBS). cDNA levels obtained using avian myeloblastosis virus (AMV) reverse transcriptase (RT) were at least twice as high as those obtained with Sensiscript and Superscript RT. RNAlater maintained RNA integrity in whole renal cortex stored at 4 degrees C for up to 3 months. Dissection of small biopsies in RNAlater rendered similar RNA yields in comparison with dissection in PBS, but the yield of glomeruli from the cortices was 50% lower (P < 0.005). Integrity of RNAlater-treated tissue, evaluated by light microscopy and immunofluorescence, was diminished. CONCLUSION:This study shows optimization of several steps in the protocol for extraction and handling of RNA in renal cortical tissue. RNA extraction and cDNA synthesis can be optimized by the use of the Trizol method and AMV RT, respectively. RNAlater is beneficial for preserving RNA integrity in whole renal cortex during storage and processing, but is not suitable for implementation in routine diagnostic histologic stainings combined with RNA expression studies in dissected biopsy material.
journal_name
Kidney Intjournal_title
Kidney internationalauthors
Roos-van Groningen MC,Eikmans M,Baelde HJ,de Heer E,Bruijn JAdoi
10.1111/j.1523-1755.2004.00366.xsubject
Has Abstractpub_date
2004-01-01 00:00:00pages
97-105issue
1eissn
0085-2538issn
1523-1755pii
S0085-2538(15)49680-2journal_volume
65pub_type
杂志文章abstract::Excess fatty acids accompanied by triglyceride accumulation in parenchymal cells of multiple tissues including skeletal and cardiac myocytes, hepatocytes, and pancreatic beta cells results in chronic cellular dysfunction and injury. The process, now termed lipotoxicity, can account for many manifestations of the 'meta...
journal_title:Kidney international
pub_type: 杂志文章,评审
doi:10.1038/sj.ki.5001834
更新日期:2006-11-01 00:00:00
abstract::Crim1 is a cell-surface, transmembrane protein that binds to a variety of cystine knot-containing growth factors, including vascular endothelial growth factor A. In the developing renal glomerulus, Crim1 acts to tether vascular endothelial growth factor A to the podocyte cell surface, thus regulating its release to gl...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.2009.345
更新日期:2009-12-01 00:00:00
abstract::Autosomal dominant polycystic kidney disease (ADPKD) is an inherited, progressive nephropathy accounting for 4-10% of end stage renal disease worldwide. PKD1 and PKD2 are the most common disease loci, but even accounting for other genetic causes, about 7% of families remain unresolved. Typically, these unsolved cases ...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1016/j.kint.2019.08.038
更新日期:2020-02-01 00:00:00
abstract::Diagnosis and treatment of atherosclerotic renal artery stenosis (ARAS) in the elderly is a never-ending challenge for every nephrologist. Kalra et al. report that adjusted hazard ratios for ARAS increased threefold from 1992 to 2004 in 16 million United States Renal Data System participants aged 66 years or older. Ho...
journal_title:Kidney international
pub_type: 信件
doi:10.1038/ki.2009.326
更新日期:2010-01-01 00:00:00
abstract::Medial arterial calcification involves chondrogenic/osteogenic transdifferentiation of vascular smooth muscle cells (VSMCs). Zhao et al. report that phosphate activates the mammalian target of rapamycin (mTOR) cascade in VSMCs, leading to downregulation of Klotho. Furthermore, rapamycin was shown to halt medial calcif...
journal_title:Kidney international
pub_type: 评论,杂志文章
doi:10.1038/ki.2015.223
更新日期:2015-10-01 00:00:00
abstract:BACKGROUND:Among the members of the small leucine-rich proteoglycan family, decorin, biglycan, and fibromodulin have been proposed to be potent modulators of transforming growth factor-beta (TGF-beta) activity, thereby playing an important role in the pathogenesis of fibrotic kidney diseases. Furthermore, decorin expre...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1046/j.1523-1755.2000.00317.x
更新日期:2000-10-01 00:00:00
abstract:BACKGROUND:Variant forms of lysozyme, a ubiquitous bacteriolytic enzyme, are known to lead to hereditary non-neuropathic renal amyloidosis and, so far, three different mutations of the lysozyme gene have been reported. In this study, we report a novel lysozyme variant, Phe57Ile, associated with renal amyloidosis in thr...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1046/j.1523-1755.2003.00904.x
更新日期:2003-05-01 00:00:00
abstract::The essential role of the glycoprotein hormone erythropoietin (Epo) in the control of red blood cell production is well established. Synthesis of Epo is induced in response to low oxygen (hypoxia). In response to stimulation, increases in serum Epo levels are paralleled by changes in the abundance of Epo mRNA. These c...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1997.70
更新日期:1997-02-01 00:00:00
abstract::The renal effects of a single intravenous dose of two different E. coli lipopolysaccharides (LPS 0111:B4 and LPS 0127:B8), at the same dose of 100 micrograms/kg, were evaluated in euvolemic Munich-Wistar (MW) rats by whole kidney clearance techniques and micropuncture studies. Following LPS infusion, a significant dec...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1989.232
更新日期:1989-10-01 00:00:00
abstract::Compelling evidence indicates that vascular endothelial growth factor (VEGF) is a fundamental regulator of normal and abnormal angiogenesis. The loss of a single VEGF allele results in defective vascularization and early embryonic lethality. VEGF plays also a critical role in kidney development, and its inactivation d...
journal_title:Kidney international
pub_type: 杂志文章,评审
doi:10.1046/j.1523-1755.1999.00610.x
更新日期:1999-09-01 00:00:00
abstract::Spleen tyrosine kinase (SYK) is an important component of the intracellular signaling pathway for various immunoreceptors. Inhibition of SYK has shown promise in preclinical models of autoimmune and glomerular disease. However, the description of SYK expression in human renal tissue, which would be desirable ahead of ...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.2015.29
更新日期:2015-07-01 00:00:00
abstract::Major temporal changes in the case mix of patients accepted for renal replacement therapy impede analysis of long-term mortality trends in end-stage renal disease patients. A record linkage study of 2 large electronic hospital inpatient data sets with national mortality data used standardization by age, sex, and comor...
journal_title:Kidney international
pub_type: 评论,杂志文章
doi:10.1016/j.kint.2018.01.002
更新日期:2018-05-01 00:00:00
abstract::The thick ascending limb of Henle's loop ( TAL ) develops a specific and consistent structural lesion during perfusion of the isolated rat kidney that progresses from mitochondrial swelling at 15 min to complete cellular disruption at 90 min. Because it seemed possible that damage localized to this metabolically activ...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1984.9
更新日期:1984-01-01 00:00:00
abstract::Recent linkage analyses of nondiabetic African-American patients with focal segmental glomerulosclerosis (FSGS) have identified MYH9, encoding nonmuscle myosin heavy chain IIA (NMMHC-IIA), as a gene having a critical role in this disease. Abnormalities of the MYH9 locus also underlie rare autosomal dominant diseases s...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.2010.21
更新日期:2010-07-01 00:00:00
abstract::The sympathetic nervous system modulates renal function through its receptors namely beta1 (cardiac output and renin release), alpha1 (systemic and renovascular constriction), and beta2 renovascular dilation. Sympathetic overactivity is commonly seen in chronic kidney disease (CKD) and is an important contributor to i...
journal_title:Kidney international
pub_type: 杂志文章,评审
doi:10.1038/sj.ki.5001835
更新日期:2006-12-01 00:00:00
abstract::Migration of monocytes into the vessel wall contributes to the onset and progression of atherosclerosis. Because monocytes are a heterogeneous population, we determined potential associations between monocyte subsets and cardiovascular events in a prospective cohort of 94 dialysis patients followed for 35 months. The ...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/sj.ki.5002744
更新日期:2008-03-01 00:00:00
abstract::We studied the specific binding of radiolabeled bradykinin ([3H]BK) and vasopressin ([3H]AVP) to membrane preparations of bovine and porcine kidney medulla. [3H]BK reversibly labeled a single site (Kd = 1.06 nM) in bovine kidney medulla independently of [Mg2+]. The number of BK receptors in bovine kidney medulla, Bmax...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1996.353
更新日期:1996-08-01 00:00:00
abstract::Clinical studies have demonstrated a poor correlation between localization tests, which are designed to determine the site of urinary tract infection, and symptoms of upper urinary tract infection. One explanation is that microorganisms may be present in the kidney but not initiate an inflammatory response with associ...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1992.200
更新日期:1992-05-01 00:00:00
abstract::Prostaglandin E2 (PGE2) may be an important negative feedback modulator of endothelin-1 (ET-1)-stimulated mesangial cell proliferation and contraction. Recent studies suggest that ET-1 may induce prolonged mesangial cell PGE2 production, however the mechanism of this effect is unknown. The current study was undertaken...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1995.6
更新日期:1995-01-01 00:00:00
abstract:BACKGROUND:Gene expression profiling of nephropathies may facilitate development of diagnostic strategies for complex renal diseases as well as provide insight into the molecular pathogenesis of kidney diseases. To test molecular based renal disease categorization, differential gene expression profiles were compared be...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1111/j.1523-1755.2004.00499.x
更新日期:2004-03-01 00:00:00
abstract::A controlled trial of cyclosporine in patients diagnosed with progressive membranous nephropathy (MGN) was carried out to determine whether cyclosporine (D) would be more effective than placebo (P) in reducing the rate of deterioration in renal function. Patients (N = 64) with MGN were placed on a restricted protein d...
journal_title:Kidney international
pub_type: 临床试验,杂志文章,随机对照试验
doi:10.1038/ki.1995.161
更新日期:1995-04-01 00:00:00
abstract::Potassium transport in the thick ascending limb is conducted by heterogeneous cells to opposite directions. Effects of glucocorticoids and mineralocorticoids on potassium transport of the rat medullary thick ascending limb (MTAL) were examined by in vitro microperfusion technique by measuring net potassium flux (JK), ...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1995.122
更新日期:1995-03-01 00:00:00
abstract::Enalapril effectively decreases hematocrits in patients with postrenal transplant erythrocytosis (PTE). We studied the effect of enalapril withdrawal on erythropoiesis in 18 patients with PTE who had been treated for 13 +/- 8 months. Hematocrit, reticulocyte count, plasma erythropoietin, and plasma insulin-like growth...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1994.411
更新日期:1994-11-01 00:00:00
abstract::The soluble urokinase receptor (suPAR) promotes proteinuria and induces focal segmental glomerulosclerosis (FSGS)-like lesions in mice. A serum suPAR concentration cutoff of 3000 pg/ml has been proposed as a clinical biomarker for patients with FSGS. Interestingly, several studies in patients with glomerulopathy found...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.2013.505
更新日期:2014-03-01 00:00:00
abstract:BACKGROUND:It is still unclear whether age per se is associated with preservation of renal functional reserve, that is, of the increase in glomerular filtration rate (GFR) induced by appropriate vasodilating stimulus. METHODS:To gain insights into this issue, we evaluated the renal response to a maximal vasodilating s...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1046/j.1523-1755.2001.0590031052.x
更新日期:2001-03-01 00:00:00
abstract::This study examined the consequences of nephrectomy in United States Army personnel who lost a kidney due to trauma during World War II (WWII). Records of 62 servicemen who underwent nephrectomy at an average age of 25 years were obtained. Mortality was compared with that of WWII servicemen of the same age. Medical re...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1993.156
更新日期:1993-05-01 00:00:00
abstract::Recent evidence suggests that glomerular hypertrophy is a key event in the development of focal and segmental glomerulosclerosis and hyalinosis (FSGS) in humans and in many experimental models of FSGS. The initial aim of the present study was to determine if glomerular hypertrophy occurs in a puromycin aminonucleoside...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1996.375
更新日期:1996-09-01 00:00:00
abstract::Carefully age-matched, purebred male beagle dogs that underwent uninephrectomy one month after they were made diabetic with alloxan were used to establish a model of rapidly developing diabetic nephropathy in a large animal. The diabetic animals, all requiring insulin, were divided into two groups: one group with cont...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1982.88
更新日期:1982-05-01 00:00:00
abstract::The release of cytokines and prostaglandins (PG) by peritoneal macrophages (PM luminal diameter of) may influence the cytokine network controlling peritoneal inflammation and in the long-term the function of the peritoneum as a dialysis membrane. In the present study, an evaluation of the long-term effects of peritone...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1996.75
更新日期:1996-02-01 00:00:00
abstract::To determine whether the induction of immune-mediated glomerular injury influences the formation of cyclooxygenase products by glomerular cells, we determined prostaglandin E2 (PGE2) and thromboxane B2 (TXB2) (as the stable metabolite of TXA2) formation in isolated glomeruli of rats with passive Heymann nephritis (PHN...
journal_title:Kidney international
pub_type: 杂志文章
doi:10.1038/ki.1987.118
更新日期:1987-05-01 00:00:00