Magnetic resonance-based visualization of thermal ablative margins around hepatic tumors by means of systemic ferucarbotran administration before radiofrequency ablation: animal study to reveal the connection between excess iron deposition and T2*-weighted hypointensity in ablative margins.

OBJECTIVE: The objective of this study was to demonstrate experimentally that radiofrequency ablation (RFA) of ferucarbotran-accumulated healthy liver tissues causes excess iron deposition in the ablated liver tissues on postablation days and produces sustained T2*-weighted low signals indicative of ablative margins surrounding hepatic tumors. MATERIALS AND METHODS: We conducted 3 experiments using 30 rats. In experiment 1, we administered either ferucarbotran (n = 6) or saline (n = 4), acquired T2*-weighted images (T2*-WIs) of the liver by using a 3-T magnetic resonance scanner, and subsequently performed RFA of healthy liver lobes. We acquired follow-up T2*-WIs up to day 7 and histologically analyzed the liver specimens. In another 4 rats, we performed sham operation, instead of RFA, in ferucarbotran-accumulated liver lobes, followed by the same image acquisition and histological analysis. In experiment 2, we administered 59Fe-labeled ferucarbotran, subsequently performed either RFA (n = 4) or sham operation (n = 4) in the liver, and acquired autoradiograms of the liver specimens on day 7. In experiment 3, we conducted RFA treatment for 8 rats bearing orthotopic hepatic tumors after ferucarbotran administration and monitored tumor growth by using serial T2*-WIs. RESULTS: On days 4 and 7 of the experiment 1, T2*-WIs of 6 rats with systemic ferucarbotran administration and subsequent hepatic RFA showed low-signal regions indicative of ablated liver tissues, whereas high-signal areas were seen in 4 saline-administered rats. Neither high nor low signal areas were detected in 4 sham-operated rats. Histologically, larger amounts of iron were observed in the RFA-induced necrotic liver tissues in the ferucarbotran-administered rats than in the saline-administered-rats. The 59Fe autoradiography of the rats in experiment 2 revealed accumulation of ferucarbotran-derived iron in necrotic liver tissues. Among 6 hepatic tumors grown in 6 rats of the experiment 3, a total of 4 tumors were stable in size, but the other 2 increased markedly on day 7. Retrospectively, T2*-WIs showed the former tumor sites surrounded completely by low-signal areas on day 4. CONCLUSIONS: The RFA of ferucarbotran-accumulated healthy liver tissues in the rats caused excess iron deposition in the ablated liver tissues and produced sustained T2*-weighted hypointense regions. Similar hypointense regions surrounding hepatic tumors were indicative of ablative margins.

Poor mental health associated with job dissatisfaction among school teachers in Japan.

School teaching is regarded as a stressful occupation. The present study aimed to compare the likelihood of having minor psychiatric disorders (MPD) among school teachers with that among civil servants, and to investigate what factors were specifically associated with MPD in teachers. We conducted a questionnaire-based survey of 403 teachers employed at state schools and 611 civil servants as a comparison group in a medium-sized city in Japan. The response rate was 59.6% for teachers and 62.0% for civil servants. Mental health was assessed using the 28-item General Health Questionnaire (GHQ-28), according to which those with a score of six or higher were considered to have MPD. Logistic regression analysis was used to identify the factors associated with MPD. Although the proportion of subjects with MPD among teachers was greater than that among civil servants, the difference in the proportion was not statistically significant in the multiple logistic regression analysis adjusted for potential confounders. In a separate analysis of the teachers, reduced job satisfaction and shorter time spent of leisure were significantly associated with an increased likelihood of having MPD. In the group of civil servants, longer working hours, reduced life satisfaction, a history of sick leave, and physical illness were associated with an increased likelihood of having MPD. When this analysis was conducted separately for male and female teachers, job dissatisfaction alone was associated with MPD only in female teachers. Poor mental health of Japanese school teachers, female teachers in particular, was found to be associated with job dissatisfaction.

Primary aldosteronism combined with preclinical Cushing's syndrome in an elderly patient.

Pre-clinical Cushing's syndrome (pre-CS) is sometimes seen with adrenal cortical tumors. An 80-year-old woman had severe hypertension and hypokalemia, the typical clinical features of primary aldosteronism, but detailed hormonal examinations revealed the accompanying pre-CS. After adrenalectomy by laparoscopy, her blood pressure was remarkably reduced and the hypokalemia also recovered. The tumor consisted of mostly light clear cells and scattered dark compact cells resembling islands. Abundant expression of cytochrome P450 aldosterone synthase in the clear cells and cytochrome P450 11beta-hydroxylase in the dark cells was detected by immunohistochemical studies, which confirmed that clear cells can produce aldosterone and compact cells can produce cortisol.

[Survey of patients with intractable diseases in Kyoto city].

OBJECTIVE: This study was performed to assess changes over a 5-year period in the number of patients with intractable diseases living in Kyoto city who received public financial aid for treatment. Variation in the quality of their lives was also investigated. METHODS: Questionnaires were mailed to all patients with intractable diseases who lived in Kyoto city and who applied for financial aid for treatment in 1996 and in 2001. RESULTS: 1. The total number of patients increased 1.4-fold over the 5 years (from 4,097 to 5,891). 2. The number of patients who required medical treatments increased. Especially a considerable number of patients required treatments for secondary conditions ascribable to a long-term bed-confined state or prolonged treatment for primary diseases. 3. The number of patients who required care to support daily life or hospital visits increased 2-fold. 4. It was demonstrated that 52.8% of patients felt their lives had improved after the introduction of long-term care insurance system. CONCLUSIONS: The number of patients with intractable diseases appears to be increasing and their clinical courses are becoming chronic and more severe. This situation can be expected to persist in the near future, so that further consideration of measures to provide medical care and welfare is necessary.

Insulin-mediated regulation of the endothelial renin-angiotensin system and vascular cell growth.

OBJECTIVE: Insulin has a growth-stimulating effect for vascular tissue. At the tissue level, the vascular renin-angiotensin system (RAS) may be involved in the progression of atherosclerosis or vascular hypertrophy. We previously reported that the vascular RAS activity is activated in vascular smooth muscle cells (SMC) by insulin stimulation. However, the effect of insulin on the RAS in endothelial cells (EC) is not fully understood. METHODS: Cultured human EC were incubated with or without insulin. After incubation for 48 h, cellular angiotensinogen and renin mRNA expression and levels in the cells were quantified by slot-blot hybridization and radioimmunoassay. Angiotensin I converting enzyme (ACE) activity in EC homogenates was measured by modified Cushman and Cheung method. EC growth and SMC with or without EC using co-culture were assessed by 3H-thymidine uptake for evaluation of their growth. RESULTS: All doses of insulin (10, 100, 1000 microU/ml) decreased angiotensinogen and renin mRNA expression (angiotensinogen: 19.3%, P < 0.05; 25.4%, P < 0.01; 26.2%, P < 0.01, renin: 12.9%, P < 0.05; 21.3%, P < 0.01; 14.3%, P < 0.05, respectively). Both cellular angiotensinogen and renin level were also reduced by high levels of insulin. Neither 10 nor 100 microU/ml insulin increased cellular angiotensin converting enzyme (ACE) activity (2.17 to 3.48-folds, P = 0.077, 0.125, respectively) significantly, but 1000 microU/ml insulin strongly up-regulated ACE activity by 16.67-folds (P = 0.001) in cultured EC. For the co-culture with EC and SMC, 100 microU/ml insulin was not able to induce SMC but 1000 microU/ml insulin accelerated SMC growth in the co-culture. In contrast insulin that was over 100 microU/ml induced SMC growth in the sole culture of SMC. CONCLUSION: Either low or high levels of insulin suppressed angiotensinogen and renin expression, however, high doses of insulin stimulated ACE activity in cultured human aortic EC. This may indicate that insulin regulates vascular cell growth and endothelial function via bifunctional modification of the vascular angiotensin generation.

Role of endothelin-1 induced by insulin in the regulation of vascular cell growth.

Insulin is not only a growth factor for vascular cells, but also an inducer of other vasoactive substances such as endothelin-1 (ET-1) in vascular cells. The aim of the present study was to assess the role of endothelial cells (EC) in insulin mediated vascular smooth muscle cell (VSMC) proliferation. Cultured human aortic EC and VSMC were separately incubated. EC were stimulated with insulin (0 to 1000 microU/mL) for 24 h, in the presence or absence of anti-insulin-growth factor-1 (anti-IGF-1) receptor antibody (alphaIR(3)) or a nonselective ET-1 receptor antagonist (TAK044). Cell proliferation was measured by determining (3)H-thymidine uptake. Although 10 microU/mL insulin did not affect ET-1 production in the EC culture medium, a higher concentration of insulin stimulated it. Production of ET-1 in EC was activated by insulin via the IGF-1 receptor, inasmuch as alphaIR(3) blocked insulin mediated upregulation of ET-1. There was no significant difference in (3)H-thymidine incorporation in the presence of insulin (up to 1000 microU/mL) or TAK044. Culture medium from EC stimulated with insulin enhanced VSMC proliferation, which was almost totally suppressed by TAK044. Insulin induced VSMC growth dose dependently when VSMC were cultured alone. In contrast, insulin at concentrations of 100 microU/mL or lower failed to stimulate growth of co-cultured VSMC, but only at 330 microU/mL or higher concentrations stimulated VSMC growth in this system. Of interest, VSMC proliferation was greatest when L-NAME was added and co-cultured with EC. In summary, a severely hyperinsulinemic state may regulate VSMC and EC proliferation via activation of vasoactive substances such as ET-1 and nitric oxide induced by insulin.

Upregulation of cAMP is a new functional signal pathway of Klotho in endothelial cells.

We measured angiotensin I-converting enzyme (ACE) activity in a human endothelial cell to characterize the intracellular signal pathways of Klotho. COS-1 cells transfected with naked mouse membrane-form klotho plasmid DNA (pCAGGS-klotho) translated proper Klotho protein. This translated Klotho protein was secreted into the culture medium. Furthermore, ACE activity in human umbilical vein endothelial cells (HUVEC) was upregulated when HUVEC were co-cultured with COS-1 cells that were pre-transfected with pCAGGS-klotho. The conditioned medium from COS-1 cells pre-transfected with pCAGGS-klotho also dose-dependently upregulated ACE in HUVEC. In addition, the conditioned medium induced time- and dose-dependent enhancement of cAMP production in HUVEC. Rp-cAMP, an inhibitor of cAMP-dependent protein kinase A (PKA), inhibited the upregulation of ACE by Klotho protein. Our results suggest that mouse membrane-form Klotho protein acts as a humoral factor to increase ACE activity in HUVEC via a cAMP-PKA-dependent pathway. These findings may provide a new insight into the mechanism of Klotho protein.

Hypertension attenuates the efficacy of hypoglycemic therapy for preserving coronary flow reserve in patients with type 2 diabetes.

Impairment of coronary flow reserve (CFR) in patients with type 2 diabetes has been generally demonstrated; however, there have been few studies investigating CFR in cases of relatively well-controlled diabetes, in distinction to the influence of hypertension. The purpose of the present study was to evaluate the influence of diabetes and hypertension upon CFR in relatively well-controlled patients. This study included 12 healthy controls (C group) and 57 patients with type 2 diabetes (DM) and/or essential hypertension who were divided into three groups as follows: patients with DM (DM group; n = 24), patients with essential hypertension (HT group; n = 15), and patients with both DM and essential hypertension (DM+HT group; n = 18). We excluded patients with evidence of coronary artery disease and/or left ventricular hypertrophy. We performed transthoracic Doppler recording of diastolic coronary flow velocity (CFV) in the left anterior descending coronary artery at rest and after maximal vasodilation by adenosine infusion (140 microg/kg/min for 3 min) CFR was defined as the ratio of hyperemic to averaged basal peak CFV. The CFR (2.92 +/- 0.46) of the DM group was not decreased compared to that of the C group (2.96 +/- 0.58), although the CFR of the HT (2.33 +/- 0.25) and DM+HT (2.35 +/- 0.25) groups were significantly reduced. Left ventricular mass index, relative wall thickness, and diastolic function were worse in the HT and DM+HT groups than in the C and DM groups. Subjects with concentric left ventricular remodeling had a lower CFR than those with normal left ventricular geometry. In conclusion, adequate hyperglycemic control prevented the progression of coronary microcirculatory disturbance, but concomitant hypertension attenuated the effect.

The renin-angiotensin and adrenergic nervous system in cardiac hypertrophy in fructose-fed rats.

BACKGROUND: Hyperinsulinemia and insulin resistance are associated with left ventricular hypertrophy (LVH) and cardiovascular complications in hypertensive subjects. The aim of this study was to explore the mechanisms for LVH including activation of the renin-angiotensin system system (RAS) and the sympathetic nervous system and their activation by insulin using a rat model of hyperinsulinemia and insulin resistance. METHODS: Male Sprague-Dawley rats were fed a high-fructose or control diet. The fructose-fed rats (FFR) were divided into four subgroups that were administrated either vehicle or the following antihypertensive drugs (n = 6-8) for 4 weeks: 1) olmesartan, an angiotensin II type 1 (AT1) receptor antagonist; 2) bunazosin, an alpha1-receptor blocker; and 3) hydralazine, a direct vasodilator. RESULTS: Fructose feeding induced significant increases in mean systolic blood pressure (BP) levels at 4 weeks (control, 117 v fructose, 131 mm Hg), left ventricular weight, and the sum of the insulin level in response to a glucose tolerance test (2 g/kg). Fructose feeding also increased urinary excretion of epinephrine and norepinephrine, the density of cardiac alpha1-adrenergic receptors, and the content of angiotensin II in the left ventricle. All antihypertensive drugs decreased systolic BP, but only the AT1 receptor antagonist attenuated the development of LVH in FFR. The AT1 receptor antagonist did not affect glucose-mediated insulin responses, but did suppress urinary catecholamine excretion and cardiac alpha1-adrenergic receptor density. CONCLUSIONS: Left ventricular hypertrophy in FFR may be less dependent on systemic elevations of BP and more dependent on the RAS and the sympathetic nervous system. Use of an AT1 receptor antagonist might be the most beneficial way to prevent progression of LVH through direct effects on tissue RAS and the sympathetic nervous system in FFR. As these changes occur in a rat model with hyperinsulinemia, insulin may have a role in promoting LVH by activating the local RAS and sympathetic nervous system activity.