Involvement of iron-evoked oxidative stress in smoking-related endothelial dysfunction in healthy young men.

BACKGROUND: Oxidative stress and smoking contribute to endothelial dysfunction. Iron might also play a role in oxidative stress generation and endothelial dysfunction. However, the involvement of iron in smoking-induced endothelial dysfunction in healthy smokers remains unclear. Therefore, we examined here whether (1) intravenous iron infusion impaired endothelial function evaluated by flow-mediated vasodilatation (FMD) in non-smokers, and (2) deferoxamine, a potent iron chelator, ameliorated endothelial dysfunction in healthy smokers. METHODS: Eight healthy young male non-smokers (23 ± 4 years old) received intravenous injection of saccharated ferric oxide (0.7 mg/kg body weight), while 10 age-matched healthy male smokers received deferoxamine mesylate (8.3 mg/kg body weight). At baseline, 5 and 20 minutes after treatment with iron or deferoxamine, biochemical variables were measured, including serum iron and marondialdehyde (MDA), a marker of lipid oxidation, and endothelial function was simultaneously evaluated by FMD. RESULTS: Compared with non-smokers, FMD was significantly lower in smokers. Iron and MDA levels were significantly increased, whereas FMD was impaired by iron infusion in non-smokers. Conversely, deferoxamine treatment significantly decreased iron and MDA levels and restored the decreased FMD in smokers. Baseline serum iron and MDA levels in all 18 subjects (non-smokers and smokers) were correlated with each other. There was a significant inverse correlation between the changes in MDA values and FMD from baseline in 18 men. Endothelium-independent vasodilation by glyceryl trinitrate was unaltered by either treatment. CONCLUSIONS: Our present study suggests that iron-evoked oxidative stress might play a role in endothelial dysfunction in healthy smokers.

In vivo identification of sentinel lymph nodes using MRI and size-controlled and monodispersed magnetite nanoparticles.

PURPOSE: To develop a sentinel lymph node (SN) identification method using accurately synthesized magnetic nanoparticles (MNPs), as an enhanced specific SN tracer in combination with magnetic resonance imaging (MRI) in intact rodent and SN metastasis models. MATERIALS AND METHODS: Three sizes of MNPs were originally synthesized. We developed an experimental rat SN model, with brachial lymph nodes (Br) as the SN and the axillary lymph node (Ax) as the second lymph node, and injection of MNPs via the front paw. SN detectability was evaluated in vivo using T1 -weighted MR images after injection of the synthesized MNPs, and the amount of iron in the Br and in the Ax was assessed using inductively coupled plasma optical emission spectrometry. RESULTS: The highest ratios of the amount of iron in the Br versus the Ax were 3.1 and 3.3, using 20-nm MNPs after 2- and 24-hour injections. The appropriate dose and particle diameter for MRI detection was optimized, and the SN was optimally distinguished in the normal and metastatic rat models using MRI after a 0.4 mg/kg 20-nm MNP injection. CONCLUSION: We developed and optimized a useful SN identification method using MRI in rodent models.

Risk factors for postoperative recurrence in patients with pathologically T1 colorectal cancer.

BACKGROUND: The evolution of diagnostic procedures has resulted in an increase in early detection of pathologically T1 (pT1) colorectal cancer (CRC). However, the risk factors affecting long-term outcomes of patients with pT1 CRCs have been unclear. The aim of the present study was to identify risk factors for postoperative recurrence and overall survival in patients with pT1 CRC. METHODS: Between January 1990 and January 2003, a total of 284 patients with pT1 CRC underwent radical surgery in the authors' institution. The impact of clinicopathological factors on postoperative recurrence and overall survival was estimated by univariate and multivariate analysis. RESULTS: The median follow-up period was 55 months (interquartile range: 47.1 months). Postoperative recurrence occurred in 8 (2.8%) patients. The overall 5-year and 10-year disease-free survival rates were 98.4 and 92.7%. Multivariate analysis showed the presence of lymphatic invasion only was an independent risk factor for postoperative recurrence in pT1 CRC patients (hazard ratio: 11.622; P = 0.003). The 5-year and 10-year disease-free survival rates of the patients in N-ly- group, the N-ly + group, and the N+ group were 99.5%/98.2% and 96.3%/75.2%, and 93.3%/93.3%, respectively. Additionally, 4 of the 8 recurrences were found more than 5 years after the operation. CONCLUSIONS: Lymphatic invasion was an independent risk factor for recurrence in pT1 CRC patients.

Mesangiolytic glomerulopathy after radiotherapy and chemotherapy of gastric lymphoma.

Mesangiolytic glomerulopathy is an uncommon complication of irradiation and chemotherapy of THP-COP [pirarubicin, cyclophosphamide (CPA), vincristin (VCR), predonisolone (PSL)] and CHOP (CPA, Doxorubicin, VCR, PSL). We report a case of 63-year-old man 7 months status post radiation, and 10 months post chemotherapy for gastric lymphoma. The patient showed proteinuria and mild renal insufficiency. Renal biopsy revealed marked mesangiolysis in the glomeruli without any immune depositions. After the administration of angiotensin II receptor blocker, the patient's renal function remained stable for over two years. Mesangiolysis was thought to be a characteristic glomerular lesion in this patient treated with both chemoagents and radiation.

Elevated circulating oxidized LDL levels in Japanese subjects with the metabolic syndrome.

In the present study, we examined the relationship between circulating oxidized low-density lipoprotein (LDL) and the metabolic syndrome in Japanese patients. Subjects who had no histories of coronary or peripheral artery disease and were taking no medications (n=119; age 57+/-10 years; male/female, 90:29) underwent a complete history and physical examination, determination of blood chemistries and oxidized LDL levels. In stepwise regression analysis, triglycerides (p=0.0001) and HDL-cholesterol (p=0.0493, inversely) were independently correlated to oxidized LDL levels. Furthermore, a significant association (p<0.0001) was found between circulating oxidized LDL levels and the accumulation of the number of the components of the metabolic syndrome. Oxidized LDL levels were one of the independent determinants of intima-media thickness of the common carotid artery, a surrogate marker of atherosclerosis. The present study reveals that circulating oxidized LDL levels are strongly associated with the metabolic syndrome. Our results suggest that elevation of oxidized LDL may be a possible molecular link between accelerated atherosclerosis and the metabolic syndrome in Japanese subjects.

Carbonic-adsorbent AST-120 reduces overload of indoxyl sulfate and the plasma level of TGF-beta1 in patients with chronic renal failure.

BACKGROUND: We previously reported a significant increase in plasma TGF-beta1 in patients with chronic renal failure (CRF). Progression of CRF may be caused by persistent renal production of TGF-beta1. In CRF rat models, an oral carbonic absorbent (AST-120) reduces the expression of the TGF-beta1 gene in the kidney, and delays the progression of CRF, in part by alleviating the overload of indoxyl sulfate. The aim of this study was to evaluate the effect of AST-120 on plasma levels of indoxyl sulfate and TGF-beta1 in CRF patients. METHODS: Ten CRF patients (aged 59.3 +/- 9.5 years, 5 men, serum creatinine 4.37 +/- 1.72 mg/dl) were enrolled in this study. All patients maintained a regular dietary therapy and the same medication throughout the study. AST-120 was added at a dose of 6 g/day. Parameters including the slope of the reciprocal of the serum creatinine-time plot, plasma indoxyl sulfate level, and plasma and urinary levels of TGF-beta1 were compared before and after the treatment with AST-120. The mean observation periods before and after the treatment were 9.7 +/- 2.8 and 6.5 +/- 2.9 months, respectively. RESULTS: Administration of AST-120 significantly reduced the plasma levels of indoxyl sulfate (1.42 +/- 1.50 vs. 1.26 +/- 1.40 mg/dl, P < 0.05) and TGF-beta1 (17.9 +/- 7.2 vs. 10.6 +/- 4.7 ng/ml, P < 0.05) and improved the slope of the reciprocal of serum creatinine (-0.061 +/- 0.041 vs. -0.032 +/- 0.055 dl/mg/year, P < 0.05). CONCLUSIONS: These results support the notion that indoxyl sulfate and TGF-beta1 may be involved in the progression of CRF, and that the oral adsorbent AST-120 may suppress the progression, at least in part, by reducing overproduction of TGF-beta1.

[Case of traditional herbal medicine-induced aristolochic acid nephropathy developing to end-stage renal failure].

A 48-year-old male was referred to our university hospital for severe azotemia with muscle cramp. He had been taking Chinese herbs as a traditional medicine to reduce hyperuricemia for about 9 months. Urinalysis showed trace proteinuria and hematuria without any casts. Renal glucosuria was also observed. In addition to azotemia, hyperchloremic metabolic acidosis and severe anemia were revealed. Hemodialysis was conducted and his general condition improved. A renal biopsy specimen revealed severe interstitial fibrosis and tubular atrophy with cellular degeneration. No remarkable glomerular changes were observed except for wrinkling of the basement membrane in a few glomeruli. Aristolochic acid was detected in the Chinese herbs, leading to the diagnosis of aristolochic acid nephropathy (AAN). His renal dysfunction was considered to be irreversible and he underwent maintenance hemodialysis. In Japan, AAN or Chinese herbs nephropathy decreased after an outbreak from 1995 to 2000. The public should be warned again that Chinese herbs, which are not permitted by the Japanese government, may contain aristolochic acid.

Comparison of histological findings and parathyroid scintigraphy in hemodialysis patients with secondary hyperparathyroid glands.

To determine the usefulness of parathyroid scintigraphy in histological estimation for secondary hyperparathyroidism (2HPT) using Tc-99m sestamibi or Tc-99m tetrofosmin. Tc-99m sestamibi (MIBI) and Tc-99m tetrofosmin (Tetro) parathyroid imaging following double-phase study, magnetic resonance imaging (MRI), and ultrasound were performed on 14 patients with 2HPT. All patients underwent parathyroidectomy. The uptake of two tracers in parathyroid areas was compared with the histopathologic findings. Forty-nine parathyroid glands were surgically explored and histologically proven to be hyperplastic. Of these, 42 were diagnosed with nodular type (N-type) hyperplasia, and 7 with diffuse type (D-type) hyperplasia. MIBI and Tetro parathyroid imagings detected 34 and 35 parathyroid glands, respectively. The sensitivity of MIBI was determined to be 76.2% (32/42) for N-type, and 28.6% (2/7) for D-type. The sensitivity of Tetro was determined to be 78.6% (33/42) for N-type and 28.6% (2/7) for D-type. The sensitivity of both MIBI and Tetro was significantly higher for N-type than for D-type, 76.2% (32/42) vs. 28.6% (2/7) in MIBI, P = 0.022; 78.6% (33/42) vs. 28.6% (2/7) in Tetro, P = 0.015. The sensitivity of MRI was determined to be 76.2% (32/42) for N-type and 42.9% (3/7) for D-type, and the sensitivity of ultrasound was 71.4% (30/42) for N-type and 71.4% (5/7) for D-type. There was no significant difference in the sensitivity of MRI or ultrasound between N-type and D-type. The uptake ratios of MIBI and Tetro were also greater for N-type than for D-type. The detectability of both MIBI and Tetro was greater for N-type than for D-type. Tc-99m MIBI or Tc-99m Tetro parathyroid scintigraphy therefore may be used clinically to distinguish N-type from D-type parathyroid gland hyperplasia.

Usefulness of bone uptake ratio of bone scintigraphy in hemodialysis patients.

OBJECTIVE: It is important to estimate the bone metabolism in patients with renal osteodystrophy. The methods of estimation must be noninvasive, accurate, and able to measure repeatedly. METHODS: The regions of interest on bone scintigraphy were drawn over the radius in 22 hemodialysis patients (10 males, 12 females). The bone/soft tissue ratio (B/ST ratio) was calculated for all patients. The bone soft tissue ratio of both skull (S) and radius (R) was obtained from the resultant count ratios. We investigated the correlation between intact parathyroid hormone (PTH), alkaline phosphatase (ALP) and the uptake ratios S and R. RESULTS: Intact PTH had a significantly linear correlation with R (r = 0.745, p < 0.0001) and S (r = 0.702, p = 0.0001). ALP also had a significantly linear correlation with R (r = 0.537, p = 0.009) and S (r = 0.772, p < 0.0001). CONCLUSION: The measurement of the bone soft tissue ratio of radius on bone scintigraphy was crucial for estimating renal osteodystrophy.

Polymorphonuclear leukocytes may impair endothelial function: results of crossover randomized study of lipid-lowering therapies.

OBJECTIVES: To examine whether polymorphonuclear leukocytes (PMNs) in hypercholesterolemia (HC) are activated to generate large amount of superoxide in vivo and hence impair endothelial function and, if so, whether statins, which possess anti-inflammatory properties, may restore PMN-mediated endothelial dysfunction. METHODS AND RESULTS: At baseline, subjects with HC showed impaired endothelial function (P<0.001), estimated by flow-mediated vasodilation of the brachial artery, and increased susceptibility of low-density lipoprotein (LDL) to oxidation (P<0.0001) compared with control subjects. PMNs obtained from HC produced greater amount of superoxide (P<0.0001), showed higher adhesiveness to cultured endothelial cells (HUVECs) (P<0.0001), and impaired endothelial nitric oxide synthase (eNOS) Ser1177 phosphorylation of HUVECs compared with controls (P<0.001). Crossover administration of fluvastatin or colestimide for 3 months lowered LDL to the same levels (P<0.001 for both). Endothelial function was restored (P<0.0001). LDL oxidation (P<0.0001) and superoxide release from PMNs (P<0.0001) were diminished only in fluvastatin but not in colestimide arm. Fluvastatin attenuated PMN adhesion to HUVECs (P<0.0001) and restored eNOS Ser1177 phosphorylation of HUVECs (P<0.001). CONCLUSIONS: Statins may improve endothelial function at least in part by inactivating neutrophils independently of LDL reduction. Our results raise a novel concept that polymorphonuclear leukocytes may attack endothelia and play a pivotal role in the pathogenesis of atherosclerosis.

AGEs activate mesangial TGF-beta-Smad signaling via an angiotensin II type I receptor interaction.

BACKGROUND: The renin-angiotensin system (RAS) and the accumulation of advanced glycation end products (AGEs) have been implicated in the pathogenesis of diabetic nephropathy. Whether there is a functional interaction between the RAS and AGEs in diabetic nephropathy is not known. In this study, we investigated whether AGEs could activate autocrine angiotensin II (Ang II) signaling and subsequently induce transforming growth factor-beta (TGF-beta)-Smad signaling in cultured rat mesangial cells. METHODS: The intracellular formation of reactive oxygen species (ROS) was detected using the fluorescent probe CM-H2DCFDA. Ang II was measured by radioimmunoassay. TGF-beta released into media was quantitatively analyzed in an enzyme-linked immunosorbent assay (ELISA). Smad2, p27(Kip1) (p27), fibronectin, and receptor for AGEs (RAGE) protein expression were determined by Western blot analysis. TGF-beta-inducible promoter activity was analyzed by a luciferase assay. DNA synthesis was evaluated by 5-bomo-2'-deoxyuridine (BrdU) incorporation and de novo protein synthesis was determined by [3H]leucine incorporation. RESULTS: AGEs increased intracellular ROS generation in mesangial cells, and this effect was significantly inhibited by an antiserum against RAGE. AGEs also were found to stimulate Ang II production in a time- and dose-dependent manner, which was completely prevented by an antioxidant, N-acetylcysteine (NAC). AGE-induced TGF-beta overproduction was completely blocked by candesartan, an Ang II type 1 receptor (AT1R) antagonist. Both candesartan and neutralizing antibody against TGF-beta completely prevented AGEs-induced Smad2 phosphorylation and TGF-beta-inducible promoter activity. Furthermore, AGEs were found to inhibit DNA synthesis and to stimulate de novo protein synthesis and fibronectin production in association with up-regulation of p27. All of these phenomena were completely prevented by candesartan or a polyclonal antibody against TGF-beta. CONCLUSION: The present study suggests that AGE-RAGE-mediated ROS generation activates TGF-beta-Smad signaling and subsequently induces mesangial cell hypertrophy and fibronectin synthesis by autocrine production of Ang II. This pathway may provide an important link between metabolic and haemodynamic factors in promoting the development and progression of diabetic nephropathy.