Relationship between Xanthine Oxidoreductase Redox and Oxidative Stress among Chronic Kidney Disease Patients.

Xanthine oxidase (XO), an isoform of xanthine oxidoreductase (XOR), is thought to increase the cardiovascular burden among chronic kidney disease (CKD) patients via oxidative radical production. Plasma XOR redox, which is characterized by the ratio of XO to total XOR, changes under different oxidative conditions associated with kidney dysfunction. However, the relationship between plasma XOR redox and oxidative stress (OS) is unclear. Thus, we aimed to clarify whether OS is related to XOR redox. We used the redox state of human serum albumin (HSA) as a marker to investigate the status of OS in CKD patients. HSA is composed of human mercaptoalbumin (HMA), which possesses not oxidized cysteine residues, reversibly oxidized human nonmercaptoalbumin-1 (HNA-1), and strongly oxidized human nonmercaptoalbumin-2 (HNA-2). The subjects included 13 nondialysis patients (7 males and 6 females) with varying degrees of kidney function. We found that ƒ(HMA) was negatively (R = -0.692, P = 0.0071) and ƒ(HNA-1) was positively (R = 0.703, P = 0.0058) correlated with plasma XO/XOR. ƒ(HNA-2) showed no correlation with XO/XOR (R = 0.146, P = 0.6412), indicating that plasma XOR redox is not related to the irreversible oxidation of HSA. In conclusion, plasma XOR redox is closely related to HSA redox, particularly reversible oxidation of HSA.

Oral Ferric Citrate Hydrate Associated With Less Oxidative Stress Than Intravenous Saccharated Ferric Oxide.

INTRODUCTION: A recent study suggested that orally dosed ferric citrate hydrate (FC) corrects renal anemia in patients on hemodialysis (HD), suggesting biological differences in effects of iron supplementation using different routes of administration. To address this issue, the present study compared oral FC with i.v. saccharated ferric oxide (FO) in stable HD patients. METHODS: Participants comprised 6 patients administered 3 consecutive protocols in the first HD session of the week in a fasting state: nothing given, as control (C); oral load of FC (480 mg iron), and 5 minutes of i.v. FO (40 mg iron). Iron dynamics in the body and biological impact on redox-inflammation status during the study (6 hours) were examined. RESULTS: Significant increases in serum iron and transferrin saturation were seen with both FC and FO. Regarding total iron-binding capacity as the sum of serum iron and unsaturated iron-binding capacity, no changes were found in FC, whereas significant increases were seen in FO (appearance of non-transferrin-binding iron [NTBI]), despite the lower serum iron levels in FO. Compared with C, increases were seen in serum myeloperoxidase (oxidative marker) with accompanying significant decreases in thioredoxin (antioxidant) in FO, whereas no changes were found in FC. CONCLUSION: Oral FC differs from i.v. FO in areas such as less NTBI generation and less induction of oxidative stress. The result indicates potential clinical benefits of oral FC in terms of iron supplementation for renal anemia in HD patients.

Comparison of albuminuria test and urine test strip in Japanese hypertensive patients: AVA-E study.

BACKGROUND: Albuminuria is thought to reflect generalized endothelial dysfunction. In hypertensive patients, albuminuria is related to the risk for cardiovascular disease (CVD) events. Thus, screening for albuminuria is critical for risk stratification in hypertensive patients. However, the actual state of albuminuria in Japanese patients without diabetes remains unclear due to insurance coverage. METHODS: The CLINITEK microalb creatinine test® is a urine test paper that can assess albumin excretion corrected for urine creatinine levels in only 60 seconds without any special equipment. The semi-quantitative albuminuria test and urine proteinuria test were performed on 8,181 Japanese hypertensive patients, and the clinical significance of the test was evaluated by comparison with the urine test strip method. RESULTS: Albumin creatinine ratio (ACR) < 30 mg/g creatinine, ACR 30 - 299 mg/g creatinine, and ACR ≥ 300 mg/g creatinine on the albuminuria test were present in 70.0%, 25.7%, and 4.3%, respectively, of patients with a negative urine protein test strip result. Furthermore, in patients with a negative urine protein test strip result, ACR ≥ 30 mg/g creatinine was independently associated with previous CVD (odds ratio: 1.25, 95% confidence interval: 1.00 - 1.57, p < 0.05) after adjustment for estimated glomerular filtration rate, age, sex, BMI, smoking, dyslipidemia, diabetes, and blood pressure categories on multivariate logistic regression analysis. CONCLUSIONS: We considered that urine test strip was inadequate test to evaluate albuminuria. Easy and quick albuminuria test on the CLINITEK MICROALB CREATININE TEST might be useful test to risk stratification of hypertensive patients compared to urine test strip.

Renoprotective effects of febuxostat in hyperuricemic patients with chronic kidney disease: a parallel-group, randomized, controlled trial.

BACKGROUND: Hyperuricemia is associated with the onset of chronic kidney disease (CKD) and renal disease progression. Febuxostat, a novel, non-purine, selective xanthine oxidase inhibitor, has been reported to have a stronger effect on hyperuricemia than conventional therapy with allopurinol. However, few data are available regarding the clinical effect of febuxostat in patients with CKD. METHODS: A prospective, randomized, open-label, parallel-group trial was conducted in hyperuricemic patients with stage 3 CKD. Patients were randomly assigned to treatment with febuxostat (n = 21) or to continue conventional therapy (n = 19). Treatment was continued for 12 weeks. The efficacy of febuxostat was determined by monitoring serum uric acid (UA) levels, blood pressures, renal function, and urinary protein levels. In addition, urinary liver-type fatty acid-binding protein (L-FABP), urinary albumin, urinary beta 2 microglobulin (ß2MG), and serum high sensitivity C-reactive protein were measured before and 12 weeks after febuxostat was added to the treatment. RESULTS: Febuxostat resulted in a significantly greater reduction in serum UA (-2.2 mg/dL) than conventional therapy (-0.3 mg/dL, P < 0.001). Serum creatinine and estimated glomerular filtration rate changed little during the study period in each group. However, treatment with febuxostat for 12 weeks reduced the urinary levels of L-FABP, albumin, and ß2MG, whereas the levels of these markers did not change in the control group. CONCLUSION: Febuxostat reduced serum UA levels more effectively than conventional therapy and might have a renoprotective effect in hyperuricemic patients with CKD. Further studies should clarify whether febuxostat prevents the progression of renal disease and improves the prognosis of CKD.

Home blood pressure control after the great East Japan earthquake in patients on chronic hemodialysis.

At 14:46 on 11 March 2011, northeastern Japan was struck by a major earthquake measuring 9.0 on the Richter scale (the Great East Japan Earthquake). Several reports have suggested a transient blood pressure (BP) increase after a major earthquake, but its impact on BP in chronic dialysis patients has not been reported. In a retrospective review of 25 hemodialysis patients who were residents of Koriyama City, changes in the morning home BP after the earthquake were investigated. Home systolic and diastolic BPs were significantly elevated 1 week after the earthquake (158 ± 16 mm Hg vs. 151 ± 13 mm Hg, P < 0.01, for systolic; 81 ± 13 mm Hg vs. 78 ± 11 mm Hg, P = 0.01, for diastolic). Mean home BP 1 week after the earthquake was unchanged from baseline in patients treated with sympatholytics and/or renin-angiotensin system (RAS) inhibitors. BP values returned to baseline by 4 weeks after the earthquake, but percent changes in mean BP were significantly greater even 2 weeks, 4 weeks, and 6 weeks after the earthquake in patients not treated with RAS inhibitors than in those treated with RAS inhibitors (2 weeks 7.0% ± 4.5% vs. 0.2% ± 5.0%, P < 0.01; 4 weeks 4.4% ± 5.9% vs. -1.8% ± 5.3%, P = 0.02; 6 weeks 4.6% ± 4.9% vs. -1.9% ± 3.9%, P < 0.01). On multiple regression analysis, RAS inhibitor use had an independent relationship with percentage increases in mean BP during the 6 weeks after the earthquake. Home BP was significantly increased after a major earthquake in patients on chronic hemodialysis. Prolonged deterioration of BP control after the earthquake was associated with non-use of RAS inhibitors.

Blood pressure elevation in hemodialysis patients after the Great East Japan Earthquake.

A major earthquake measuring 9.0 on the Richter scale struck northeastern Japan at 2:46 pm on 11 March 2011. Several reports have described transient increases in blood pressure after major earthquakes, but the impact of such increases on hemodialysis patients has not been reported. We retrospectively investigated changes in blood pressure and influencing factors in 205 patients (mean age 66.6±13.0 years; male 51.7%; median dialysis vintage 6.0 (2.0-11.0) years) on chronic dialysis at three dialysis centers in the affected area (Fukushima City) for 8 weeks after the earthquake. Pre-dialysis blood pressure was significantly elevated at 1 week after the earthquake compared with baseline (systolic vs. diastolic blood pressure: 153.1±20.2/80.1±13.5 vs. 148.6±20.0/77.5±12.8 mm Hg, P<0.001), similarly post-dialysis blood pressure was elevated for up to 8 weeks. Independent factors influencing changes in blood pressure after the earthquake comprised baseline blood pressure and α-blockers. The earthquake induced a significant elevation in blood pressure among patients on chronic dialysis, and activation of the sympathetic nervous system might at least in part be associated with the mechanism underlying this increase.

Skin autofluorescence is associated with the progression of chronic kidney disease: a prospective observational study.

BACKGROUND: Advanced glycation end product (AGE) accumulation is thought to be a measure of cumulative metabolic stress that has been reported to independently predict cardiovascular disease in diabetes and renal failure. The aim of this study was to evaluate the association between AGE accumulation, measured as skin autofluorescence, and the progression of renal disease in pre-dialysis patients with chronic kidney disease (CKD). METHODS: Skin autofluorescence was measured noninvasively with an autofluorescence reader at baseline in 449 pre-dialysis patients with CKD. The primary end point was defined as a doubling of serum creatinine and/or need for dialysis. RESULTS: Thirty-three patients were lost to follow-up. Forty six patients reached the primary end point during the follow-up period (Median 39 months). Kaplan-Meier analysis showed a significantly higher risk of development of the primary end points in patients with skin autofluorescence levels above the optimal cut-off level of 2.31 arbitrary units, derived by receiver operator curve analysis. Cox regression analysis revealed that skin autofluorescence was an independent predictor of the primary end point, even after adjustment for age, gender, smoking history, diabetes, estimated glomerular filtration rate and proteinuria (adjusted hazard ratio 2.58, P = 0.004). CONCLUSIONS: Tissue accumulation of AGEs, measured as skin autofluorescence, is a strong and independent predictor of progression of CKD. Skin autofluorescence may be useful for risk stratification in this group of patients; further studies should clarify whether AGE accumulation could be one of the therapeutic targets to improve the prognosis of CKD.

Hypertrophic cranial pachymeningitis in MPO-ANCA-related vasculitis: a case report and literature review.

A 75-year-old woman presented with rapidly progressive glomerulonephritis with positive results for anti-myeloperoxidase anti-neutrophil cytoplasmic antibody (MPO-ANCA). Corticosteroid therapy was successfully introduced. However, 7 months later, magnetic resonance imaging revealed marked swelling in the falx cerebri and high density regions were apparent on gallium scintigraphy, leading to diagnosis of hypertrophic cranial pachymeningitis (HCP). Symptoms improved with intensified corticosteroid therapy, but radiological examination 9 months later revealed right nasal sinus inflammation accompanied by osteolytic change. Granulomatosis with polyangiitis (Wegener's) was finally diagnosed. HCP is an important complication in MPO-ANCA-related vasculitis, and needs to be considered during the clinical course.

The clinical applicability of albuminuria testing in Japanese hypertensive patients: the AVA-E study.

OBJECTIVE: Albuminuria is thought to reflect generalized endothelial dysfunction. In hypertensive patients, albuminuria increases the risk of cardiovascular disease (CVD) events. Therefore, screening for albuminuria is critical for stratifying risks in hypertensive patients. However, a limited number of Japanese studies have performed quantitative examinations of albuminuria. The objective of this study was to examine the utility of the CLINITEK MICROALB CREATININE TEST for albuminuria screening. MATERIALS: The CLINITEK MICROALB CREATININE TEST consists of a urine test strip that assesses albumin excretion corrected for the urine creatinine levels in only 60 seconds without the need for any special facilities. The CLINITEK MICROALB CREATININE TEST was performed in 5,647 Japanese hypertensive patients, excluding diabetic patients, and the clinical significance of the test was evaluated. RESULTS: According to the CLINITEK MICROALB CREATININE TEST, the A1 (albumin creatinine ratio: ACR <30 mg/gï½¥creatinine), A2 (ACR 30-299 mg/gï½¥creatinine) and A3 (ACR ≥ 300 mg/gï½¥creatinine) levels of albuminuria were present in 61.2%, 32.5% and 6.3% of the patients surveyed, respectively. The proportions of A2 and A3 patients increased with chronic kidney disease (CKD) stage, blood pressure, age and previous history of CVD. According to a multivariate logistic regression analysis, the A2 and A3 levels of albuminuria were found to be independently associated with a previous history of CVD (odds ratio: 1.36, 95% confidence interval: 1.08-1.72, p<0.01) after adjusting for age, diabetes, blood pressure and estimated glomerular filtration rate (eGFR). CONCLUSION: In hypertensive patients, the A2 and A3 levels of albuminuria on the CLINITEK MICROALB CREATININE TEST are associated with a previous history of CVD, independent of eGFR. Therefore, by reflecting the status of systemic vascular injury, this test may help to perform CVD risk stratification.

Acute changes in home blood pressure after the Great East Japan Earthquake among patients with chronic kidney disease in Fukushima City.

BACKGROUND: Blood pressure (BP) transiently increases after major earthquakes. Although home BP increased in hypertensive patients after the Great East Japan Earthquake (measuring 9.0 on the Richter scale) on March 11th, 2011, such profiles in patients with chronic kidney disease (CKD) have not been investigated. METHODS: This retrospective single-center observational study included 38 patients with CKD at the predialysis stage [male, n=27 (71%); mean age 66.0 years; mean estimated glomerular filtration rate (eGFR) 46.0 mL/min/1.73 m²] who lived in Fukushima City. We compared their morning BP between two and four weeks after the quake with that of a control group of 39 non-CKD patients with hypertension. RESULTS: Systolic BP (SBP) remained elevated for one week after the quake in the CKD and non-CKD groups [before vs. after the quake 133.7±15.6 vs. 136.9±16.9 (p=0.017) and 129.9±7.8 vs. 133.3±9.3 mmHg (p=0.009), respectively]. Increases in SBP in the morning after the quake were statistically significant in the group with but not in that without CKD (7.1 and 3.4 mmHg; p=0.038 and 0.221, respectively). Multivariate analysis revealed that a low eGFR was an independent risk factor for elevated SBP after the quake. CONCLUSIONS: The quake caused acute changes in the home BP and the fact that BP elevation correlated with renal function suggests a possible mechanism of CKD, such as enhanced activity of the sympathetic nervous system in such patients.

Clinical outcomes of Japanese MPO-ANCA-related nephritis: significance of initial renal death for survival.

OBJECTIVE: Anti-myeloperoxidase anti-neutrophil cytoplasmic antibody (MPO-ANCA)- related nephritis constitutes 60% of rapidly progressive glomerulonephritis (RPGN) in Japan. The reported 1-year survival rate is over 80%, however, the long-term prognosis remains unknown. We therefore investigated the prognosis and factors affecting the clinical course of patients. METHODS: We retrospectively investigated 74 patients (female, n=42; median age, 73.0 years) with MPO-ANCA-related nephritis. The patients were admitted to Fukushima Medical University and two affiliated hospitals between 2000 and 2010. RESULTS: Median estimated GFR (eGFR) was 12.1 mL/min/1.73 m2 at admission. The Birmingham Vasculitis Activity Score (BVAS version 3: max 63 points) at diagnosis and at 4 weeks after start of treatment were 15.0 and 5.0, respectively. Twenty-three patients (31%) died during a median observation period of 30.5 months. Sixteen patients (22%) presented with end-stage renal disease (ESRD) at the initial phase, and needed regular dialysis therapy. Multivariate Cox proportional hazards model analysis revealed that renal death at the initial phase was a significant risk factor for all-cause death (Hazard ratio, 5.72; 95% confidence interval, 2.49-13.09; p<0.001). Furthermore, BVAS>6, evaluated 4 weeks after start of treatment, is an independent risk factor for ESRD and patient survival. CONCLUSION: This is the first investigation to demonstrate clinical features focusing on MPO-ANCA-related nephritis. Renal death at the initial phase of treatment is a powerful risk factor for all-cause death in patients with MPO-ANCA-related nephritis. Patients at high risk of death and ESRD could be stratified according to BVAS.

The great East Japan earthquake: blood pressure control in patients with chronic kidney disease.

BACKGROUND: At 1446 on 11 March 2011, northeastern Japan was struck by a major earthquake measuring 9.0 on the Richter scale. There have been several reports of transient blood pressure increases after a major earthquake, but the impact of a major earthquake on blood pressure in chronic kidney disease (CKD) patients has not been fully investigated. METHODS: Changes in clinic blood pressure following the earthquake were investigated in 132 hypertensive patients with stage 3 and 4 CKD who were residents of Fukushima City. RESULTS: Both systolic and diastolic blood pressures were significantly elevated 1-3 weeks after the earthquake compared with before the earthquake (134 ± 19 mm Hg vs. 138 ± 20 mm Hg, P = 0.02 for systolic; 76 ± 13 mm Hg vs. 79 ± 12 mm Hg, P = 0.01 for diastolic), and these values returned to baseline by 5-7 weeks after the earthquake. Multiple logistic regression analysis identified male sex (odds ratio (OR), 0.35; 95% confidence interval (CI), 0.14-0.86; P = 0.02), mean blood pressure at baseline (OR, 0.92; 95% CI, 0.86-0.96; P < 0.01), and sympatholytic medications, such as α-blockers, ß-blockers, or central sympatholytics (OR, 0.23; 95% CI, 0.07-0.76; P = 0.01), as independent factors related to elevation of mean blood pressure 1-3 weeks after the earthquake in CKD patients. CONCLUSIONS: Blood pressure was significantly increased after a major earthquake in hypertensive patients with stage 3 and 4 CKD. During the first 3 weeks after the earthquake, blood pressure control was associated with the use of sympatholytic medications.

Clinical assessment of bortezomib for multiple myeloma in comparison with thalidomide.

PURPOSE: We studied the efficacy and safety of bortezomib (BOR) for treatment of multiple myeloma in comparison with thalidomide (THAL) by reference to adverse events, and searched for laboratory markers that could be used for prognostication of patients. METHODS: Biochemical data of patients receiving BOR and THAL for treatment of multiple myeloma at the Japanese Red Cross Narita Hospital were investigated retrospectively, after obtaining Institutional Review Board approval. Judgment of curative effects complied with the effects criteria of the International Myeloma Working Group (IMWG). RESULTS: BOR showed a higher rate of effectiveness than THAL for refractory multiple myeloma, and its effects were rapid. BOR treatment prolonged the survival time of THAL-resistant patients. The efficacy of BOR was unrelated to patient age, the number of previous therapeutic regimens, or the disease period. After medication with BOR, patients in whom it had been effective tended to show an increase of the serum alkaline phosphatase (ALP) level. Thrombocytopenia (86.2%) and leucopenia (69.0%) were observed at high frequencies, but no previously unreported adverse events or fatalities were associated with BOR therapy. CONCLUSION: It is suggested that BOR has therapeutic efficacy for multiple myeloma as a first-line medical treatment and/or for patients with THAL resistance, and can improve prognosis and survival. Since serum ALP elevation was observed in many patients for whom BOR was effective, this may be a predictor of BOR efficacy.

Skin autofluorescence is associated with renal function and cardiovascular diseases in pre-dialysis chronic kidney disease patients.

BACKGROUND: Tissue accumulation of advanced glycation end-products (AGE) is thought to be a contributing factor to the progression of cardiovascular disease (CVD). Skin autofluorescence, a non-invasive measure of AGE accumulation using autofluorescence of the skin under ultraviolet light, has shown associations with CVD in haemodialysis patients. The present study aimed to evaluate relationships of skin autofluorescence to renal function as well as CVD in pre-dialysis patients with chronic kidney disease (CKD). METHODS: Subjects in this cross-sectional analysis comprised 304 pre-dialysis CKD patients [median age, 62.0 years; median estimated glomerular filtration rate (eGFR), 54.3 mL/min/1.73 m(2); diabetes, n = 81 (26.6%)]. AGE accumulation in skin was assessed by skin autofluorescence using an autofluorescence reader. Relationships between skin autofluorescence, eGFR, CVD history and other parameters were evaluated. RESULTS: Skin autofluorescence correlated negatively with eGFR (r = -0.42, P < 0.01) and increased as CKD stage advanced. Multiple regression analysis revealed significant correlations of skin autofluorescence with age, presence of diabetes, eGFR and CVD history in CKD patients (R(2) = 30%). Age, male gender, smoking history, skin autofluorescence and eGFR were significantly correlated with CVD history, and multiple logistic regression analysis identified age [odds ratio (OR), 1.09; 95% confidence interval (CI), 1.03-1.15; P < 0.01], history of smoking (OR, 6.50; 95%CI, 1.94-21.83; P < 0.01) and skin autofluorescence (OR, 3.74; 95%CI, 1.54-9.24; P < 0.01) as independent factors. CONCLUSIONS: Tissue AGE accumulation measured as skin autofluorescence increased as GFR decreased and was related to CVD history in CKD patients. Non-invasive autofluorescence readers may provide potential markers for clinical risk assessment in pre-dialysis CKD patients.

Effects of piclozotan (SUN N4057), a partial serotonin 1A receptor agonist, on motor complications induced by repeated administration of levodopa in parkinsonian rats.

Serotonin 1A receptor agonists have attracted much interest recently as potential therapeutic agents for levodopa-induced motor complications, such as dyskinesia and motor fluctuations. The effects of piclozotan (SUN N4057) on a rat model of advanced Parkinson's disease were investigated. Parkinsonian rats, unilaterally 6-hydroxydopamine-lesioned rats, were administered levodopa for 8 to 9 weeks. Based on the results of rotational behavior and forelimb hyperkinesia in Week 5, the rats were allocated to three treatment groups (saline and two dosing rates of piclozotan set at 0.018 and 0.036 mg/kg/h). Piclozotan was administered via continuous subcutaneous infusion using an osmotic pump for 3 to 4 weeks. At Week 7 of repeated levodopa dosing, the effects of piclozotan on levodopa-induced behavior were evaluated. In addition, extracellular levels of levodopa-derived dopamine in the striatum were measured using microdialysis in Weeks 8 to 9 after completion of the respective behavioral studies. Chronic treatment with levodopa-induced forelimb hyperkinesia and shortened the duration of rotational behavior. Piclozotan (0.018 and 0.036 mg/kg/h, plasma concentrations 5.3±0.7 and 14.3±2.9 ng/ml) reduced levodopa-induced forelimb hyperkinesia by 55% and 69%, respectively, at 1h relative to the control. Piclozotan (0.036 mg/kg/h) significantly lengthened the duration of rotational behavior by 26% versus the control and attenuated the increase in striatal levodopa-derived extracellular dopamine levels. These findings suggest that piclozotan, a serotonin 1A agonist, can improve motor complications in patients with advanced Parkinson's disease.

Tissue levels of pyrimidine nucleoside phosphorylase activity in human and rodent bladder cancer and normal bladder tissue.

OBJECTIVES: To assess the relationship between the tissue levels of pyrimidine nucleoside phosphorylase (PyNpase) and clinicopathological parameters in human bladder cancer and to investigate the PyNpase levels in rat and mouse urinary bladder initiated by N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN). METHODS: The PyNpase levels in tumor tissue, normal tissue adjacent to the tumor, and normal tissue apart from the tumor were measured in 102 patients. Additionally, the PyNpase levels were measured in rat and mouse urinary bladders treated with BBN. RESULT: The PyNpase levels of tumor tissue significantly correlated to the tumor grade and growth pattern (papillary/non-papillary), while stage, multiplicity, and tumor shape (peduncle/sessile) were not independent factors. The low-risk tumor of primary, single, G1-Ta showed significantly low levels of PyNpase. The PyNpase levels in the tumor tissue were significantly higher than those in the normal tissue. The PyNpase levels in the adjacent normal tissue were significantly higher than those in the distant normal tissue. The PyNpase levels in rat bladder tissue were significantly higher in the BBN-treatment groups than in those in the control group, only during the early carcinogenic stage. The PyNpase levels in mouse bladder tissue were significantly higher in BBN-treatment groups than in those in the control group during the whole experiment period. CONCLUSION: Our results indicated that not only tumor tissue but also normal tissue adjacent to the tumor had a potential of angiogenesis for tumor development, and transurethral resection of the bladder tumor with a wide normal margin seems to be a reasonable strategy for decreasing the risk of recurrence.