Development of the gateway recycling cloning system for multiple linking of expression cassettes in a defined order, and direction on gateway compatible binary vectors.

We developed the Gateway recycling cloning system, which allows multiple linking of expression cassettes by multiple rounds of the Gateway LR reaction. Employing this system, the recycling donor vector pRED419 was subjected to the first LR reaction with an attR1-attR2 type destination vector. Then conversion vector pCON was subjected to an LR reaction to restore the attR1-attR2 site on the destination vector for the next cloning cycle.

Role of leukotriene B4 in accelerated hyperlipidaemic renal injury.

Aim: Glomerular infiltration of macrophages is a characteristic alteration of renal pathology in hyperlipidaemic renal injury. Leukotriene B4 (LTB4) is a bioactive eicosanoid and macrophage and has two key enzymes for LTB4 synthesis, 5-lipoxygenase and leukotriene A4 (LTA4) hydrolase. The purpose of this study was to evaluate the role of LTB4 in accelerated hyperlipidaemic renal injury.

Development of Gateway binary vectors, R4L1pGWBs, for promoter analysis in higher plants.

We developed a new series of Gateway binary vectors for plant transformation, R4L1pGWBs, which allow easy construction of promoter:reporter clones. R4L1pGWBs contain a recombination attR4-attL1-reporter cassette, and thus an attL4-promoter-attR1 entry clone was efficiently incorporated by the Gateway LR reaction, resulting in the generation of an attB4-promoter-attB1-reporter construct. The reporters employed in R4L1pGWBs were beta-glucuronidase (GUS), luciferase (LUC), enhanced yellow fluorescent protein (EYFP), enhanced cyan fluorescent protein (ECFP), G3 green fluorescent protein (G3GFP), G3GFP-GUS, and tag red fluorescent protein (TagRFP).

Functional polymorphism of the myeloperoxidase gene in hypertensive nephrosclerosis dialysis patients.

Myeloperoxidase (MPO) may play an important role not only in host defense reactions but also in local inflammations, especially in atherosclerotic diseases such as hypertensive nephrosclerosis (HN). Paradoxically, MPO-deficient mice have been reported to show increased atherosclerosis compared with wild mice, although higher MPO levels are thought to exacerbate atherosclerotic disease. To clarify the genetic role of MPO in HN, we examined the function and distribution of the -463G/A polymorphism located in the promoter region of the MPO gene with ex vivo flow cytometry analysis and a study in end-stage renal disease patients, respectively.

Double filtration plasmapheresis can decrease factor XIII Activity.

Factor XIII (FXIII) produces cross-linkages among fibrin molecules within fibrin clots. Its deficiency is related with bleeding diathesis or retardation of wound healing. We report the possibility that intense double filtration plasmapheresis (DFPP) is associated with decreased FXIII activity.

Successful treatment by double filtrate plasmapheresis in a pregnant woman with the rare P blood group and a history of multiple early miscarriages.

Individuals of P type, a rare blood group, have anti-PP(1)P(k) antibody in their serum, which causes spontaneous abortion in the early stages. We report a patient of p type suffering from multiple spontaneous abortions. We also review previously reported cases from published work.

Attenuation of folic acid-induced renal inflammatory injury in platelet-activating factor receptor-deficient mice.

Platelet-activating factor (PAF), a potent lipid mediator with various biological activities, plays an important role in inflammation by recruiting leukocytes. In this study we used platelet-activating factor receptor (PAFR)-deficient mice to elucidate the role of PAF in inflammatory renal injury induced by folic acid administration. PAFR-deficient mice showed significant amelioration of renal dysfunction and pathological findings such as acute tubular damage with neutrophil infiltration, lipid peroxidation observed with antibody to 4-hydroxy-2-hexenal (day 2), and interstitial fibrosis with macrophage infiltration associated with expression of monocyte chemoattractant protein-1 and tumor necrosis factor-alpha in the kidney (day 14).

Functional polymorphisms in the vascular endothelial growth factor gene are associated with development of end-stage renal disease in males.

This study elucidates the genetic role of vascular endothelial growth factor (VEGF) as a predisposing factor for progression of chronic kidney disease. Single-nucleotide polymorphisms were genotyped and haplotype structures were determined in the 3' untranslated region (UTR) of VEGF gene, and the distribution of each haplotype in male patients with ESRD (n=101) and healthy male control subjects (n=189) was examined. The 936C/T and 1451C/T polymorphisms in the 3' UTR were in nearly absolute linkage disequilibrium, and haplotype analysis demonstrated that they were the primary responsible single-nucleotide polymorphisms.

Pulse total-hemoglobinometer provides accurate noninvasive monitoring.

Objective: Rapid noninvasive measurement of total hemoglobin would be extremely useful for various clinical situations. This study determined the clinical accuracy and utility for a pulse total-hemoglobinometer using four wavelengths: 660 nm (reduced hemoglobin), 805 nm (isosbestic point), 940 nm (oxygenated hemoglobin), and 1300 nm (water density).

Design: Clinical trial.

Haplotype analysis of NAD(P)H oxidase p22 phox polymorphisms in end-stage renal disease.

NAD(P)H oxidase is one of the most important sources of reactive oxygen species and has been demonstrated to be upregulated by angiotensin II in the kidney. Given the effect of angiotensin-converting enzyme inhibitors on the progression of both diabetic and non-diabetic renal disease, we hypothesized that the polymorphisms of NAD(P)H oxidase are associated with development of end-stage renal disease (ESRD). We examined five polymorphisms in the CYBA gene encoding the p22 phox component of NAD(P)H oxidase, including 242C/T and 640A/G polymorphisms in 467 ESRD patients and 490 healthy individuals.

Radical scavenger edaravone developed for clinical use ameliorates ischemia/reperfusion injury in rat kidney.

Background: Edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) is a potent scavenger of free radicals and has the antioxidant ability to inhibit lipid peroxidation. Its protective effect on brain ischemia has been shown. This study aimed to elucidate its possible therapeutic effects on renal oxidative stress in a rat ischemia/reperfusion model.

High-throughput single nucleotide polymorphism typing by fluorescent single-strand conformation polymorphism analysis with capillary electrophoresis.

In this study, we performed high-throughput and precise single nucleotide polymorphism (SNP) typing by fluorescent capillary electrophoresis single-strand conformation polymorphism (CE-SSCP) analysis. A system composed of a multicapillary DNA analyzer, a newly developed sieving matrix, four different colors of fluorescent labels, and a multiplex polymerase chain reaction (PCR) enabled low-cost and highly reliable SNP typing. Moreover, this system enabled the estimation of SNP allele frequencies using pooled DNA samples, which should be beneficial for large-scale association studies.

Effects of probucol on renal function and urinary protein excretion in spontaneously hypercholesterolemic rats fed a normal or high cholesterol diet.

Background/aim: Spontaneously hypercholesterolemic (SHC) rats develop hypercholesterolemia and focal glomerular sclerosis, and have been thought to be a model of lipid-induced glomerular injury. However, recent studies suggest that the hypercholesterolemia might be due to secondary mechanisms by massive proteinuria. The purpose of the present study was to determine in SHC rats the effects of a high cholesterol diet on serum lipid profiles and renal function/histology, and to examine whether or not the model of lipid-induced renal injury could be developed in a short period of the time.

Serum protein acrolein adducts: utility in detecting oxidant stress in hemodialysis patients and reversal using a vitamin E-bonded hemodialyzer.

Accumulating evidence indicates that protein modification by acrolein is one of the major hallmarks of atherosclerosis. The purpose of the present study was to evaluate the serum acrolein-modified protein adduct (Acr) level in end-stage renal disease (ESRD), and to elucidate the efficacy of vitamin E-bonded hemodialyzer in reducing Acr in a crossover trial. A significant increase in Acr was found in ESRD patients compared with healthy controls (p <.

Association of eNOS Glu298Asp polymorphism with end-stage renal disease.

Nitric oxide (NO) derived from endothelial cells is profoundly related to the maintenance of physiological vascular tone. Impairment of endothelial NO generation brought about by gene polymorphism is considered the major deterioration factor for progressive renal disease, including diabetic nephropathy. The present study aimed to elucidate the Glu298Asp polymorphism of endothelial NO synthase (eNOS) in patients with end-stage renal disease (ESRD) and its role as a predisposing factor for cardiovascular complications.

Quantifying porphobilinogen deaminase mRNA in microdissected nephron segments by a modified RT-PCR.

Background: Quantifying mRNA levels by reverse transcription-polymerase chain reaction (RT-PCR), although widely exercised, is still difficult.

Methods: A modified quantitative RT-PCR in which genomic DNA was used as standard was developed. The quantity of mRNA was expressed as the ratio of the PCR product from cDNA and that from genomic DNA (CG ratio).