Enhanced energy expenditure and fat oxidation in humans with high BMI scores by the ingestion of novel and non-pungent capsaicin analogues (capsinoids).

The biochemical and physiological indices were monitored in 44 subjects after 4-week capsinoids (capsaicin analogues with low pungency) intake. The subjects were randomly assigned to 3 groups: CSNs3 (3 mg/kg of capsinoids), CSNs10 (10 mg/kg of capsinoids) and the control (placebo). Measurements were performed in the morning on overnight-fasted subjects. The oxygen consumption (VO(2)), resting energy expenditure (REE) and fat oxidation increased slightly compared to pre-administration values without any adverse effects, although the increase was not significant. The increase in fat oxidation was positively and significantly correlated with the body mass index (BMI). A meta-analysis was therefore conducted on a subgroup consisting of subjects with BMI >or= 25 (n=28). As a result, not only VO(2) increased significantly (p<0.05) in the CSNs10 group, but also REE in the CSNs10 group and fat oxidation in the CSNs3 and CSNs10 groups tended to increase (p<0.1). Consequently, a capsinoids intake would be able to enhance the energy expenditure and fat burning in humans, particularly those with high BMI.

Preventive and therapeutic effects of angiotensin II type 1 receptor blocker on hepatic fibrosis induced by bile duct ligation in rats.

BACKGROUND: The aim of this study was to examine the preventive and therapeutic effects of an angiotensin II type 1 receptor blocker, candesartan, on cholestasis-induced liver fibrosis. METHODS: Candesartan was administered orally for 21 days immediately after bile duct ligation to evaluate its preventive effect, and for 21 days starting 3 weeks after bile duct ligation to evaluate its therapeutic effect. Fibrosis was assessed by measuring hepatic hydroxyproline (Hyp) content. The activated hepatic stellate cells (HSCs) were assessed by alpha-smooth muscle actin (alpha-SMA) immunostaining. The gene expression of collagen I, transforming growth factor-beta1 (TGF-beta1), and connective tissue growth factor (CTGF) in the liver was examined by real-time reverse transcriptase-polymerase chain reaction. RESULTS: As a preventive effect, candesartan reduced the hepatic Hyp content by 36%, alpha-SMA-positive cells by 65%, hepatic TGF-beta1 content by 35%, and the expression of collagen I by 72%, TGF-beta1 by 67%, and CTGF mRNA by 69%. As a therapeutic effect, candesartan reduced the hepatic Hyp content by 48%, TGF-beta1 content by 54%, and the expression of collagen I by 47%, TGF-beta1 by 43%, and CTGF mRNA by 53%. Significant decreases in lipid peroxidation markers, hepatic thiobarbituric acid-reactive substance, and 4-hydroxy-2-nonenal were observed in candesartan-treated rats. CONCLUSIONS: Candesartan attenuated liver fibrosis via suppression of collagen I and TGF-beta1 expression, HSC activation, and lipid peroxidation protein, showing its preventive and therapeutic effects on cholestasis-induced liver fibrosis.

Involvement of Raf-1/MEK/ERK1/2 signaling pathway in zinc-induced injury in rat renal cortical slices.

Zinc is an essential nutrient that can also be toxic. We have previously reported that zinc-related renal toxicity is due, in part, to free radical generation in the renal epithelial cell line, LLC-PK(1) cells. We have also shown that an MEK1/2 inhibitor, U0126, markedly inhibits zinc-induced renal cell injury. In this study, we investigated the role of an upstream MEK/ERK pathway, Raf-1 kinase pathway, and the transcription factor and ERK substrate Elk-1, in rat renal cortical slices exposed to zinc. Immediately after preparing slices from rat renal cortex, the slices were incubated in medium containing Raf-1 and MEK inhibitors. ERK1/2 and Elk-1 activation were determined by Western blot analysis for phosphorylated ERK (pERK) 1/2 and phosphorylated Elk-1 (pElk-1) in nuclear fractions prepared from slices exposed to zinc. Zinc caused not only increases in 4-hydroxynonenal (4-HNE) modified protein and lipid peroxidation, as an index of oxidant stress, and decreases in PAH accumulation, as that of renal cell injury in the slices. Zinc also induced a rapid increase in ERK/Elk-1 activity accompanied by increased expressions of pERK and pElk-1 in the nuclear fraction. A Raf-1 kinase inhibitor and an MEK1/2 inhibitor U0126 significantly attenuated zinc-induced decreases PAH accumulation in the slices. The Raf-1 kinase inhibitor and U0126 also suppressed ERK1/2 activation in nuclear fractions prepared from slices treated with zinc. The present results suggest that a Raf-1/MEK/ERK1/2 pathway and the ERK substrate Elk-1 are involved in free radical-induced injury in rat renal cortical slices exposed to zinc.

Expression of oxidative stress-related molecules in circulating leukocytes and urine in patients with chronic viral hepatitis.

AIMS: Oxidative stress plays a role in pathogenesis of chronic viral hepatitis. Expression of oxidative stress-related molecules remains to be clarified. METHODS: 4-hydroxy-2-nonenal (4-HNE), 4-hydroxy-2-hexenal (4-HHE), catalase, superoxide dismutase-1 (SOD-1), glutathione peroxidase-1, thioredoxin (TRX) in leukocytes and urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) were examined in 164 persons, including 130 chronic viral hepatitis patients and 34 normal individuals, by Western blot analysis and enzyme-linked immunosorbent assay, respectively. Hepatic expression of these proteins was immunohistochemically examined in 12 patients with chronic viral hepatitis, compared with three persons without liver damage. RESULTS: The 4-HNE/beta-actin ratios in chronic viral hepatitis were significantly higher than those in normal individuals (P<0.01), and were significantly correlated with asparate aminotransferase (AST) and alanine aminotransferase (ALT) (P<0.01, each). The catalase/beta-actin and SOD-1/beta-actin ratios in chronic viral hepatitis were higher than those in normal individuals, and were significantly correlated with 4-HNE/beta-actin ratios (P<0.01, each). Hepatic expression of 4-HNE, 4-HHE, catalase, SOD-1 and TRX in chronic viral hepatitis was higher than that without liver damage. Urinary excretion of 8-OHdG was not changed in chronic viral hepatitis. CONCLUSIONS: The results of the present study suggest that expression of oxidative stress-related molecules in leukocytes is upregulated in relation to serum aminotransferase levels.

Protective effect of serum thymic factor, FTS, on cephaloridine-induced nephrotoxicity in rats.

Serum thymic factor (FTS), a thymic peptide hormone, has been reported to increase superoxide disumutase (SOD) levels in senescence-accelerated mice. In the present study, we examined the effect of FTS on cephaloridine (CER)-induced nephrotoxicity in vivo and in vitro. We previously reported that CER led to extracellular signal-regulated protein kinase (ERK) activation in the rat kidney. So, we also investigated whether FTS has an effect on ERK activation induced by CER. Treatment of male Sprague-Dawley rats with intravenous CER (1.2 g/kg) for 24 h markedly increased BUN and plasma creatinine levels and urinary excretion of glucose and protein, decreased creatinine clearance and also led to marked pathological changes in the proximal tubules, as revealed by electron micrographs. An increase in phosphorylated ERK (pERK) was detected in the nuclear fraction prepared from the rat kidney cortex 24 h after CER injection. Pretreatment of rats with FTS (50 microg/kg, i.v.) attenuated the CER-induced renal dysfunction and pathological damage. FTS also suppressed CER-induced ERK activation in the kidney. In vitro treatment of the established cell line, LLC-PK1 cells, with FTS significantly ameliorated CER-induced cell injury, as measured by lactate dehydrogenase (LDH) leakage. Our results, taken together with our previous report that MEK inhibitors ameliorated CER-induced renal cell injury and ERK activation induced by CER, suggest that FTS participates in protection from CER-induced nephrotoxicity by suppressing ERK activation induced by CER.

Serum thymic factor, FTS, attenuates cisplatin nephrotoxicity by suppressing cisplatin-induced ERK activation.

Serum thymic factor (FTS), a thymic peptide hormone, has been reported to attenuate the bleomycin-induced pulmonary injury and also experimental pancreatitis and diabetes. In the present study, we investigated the effect of FTS on cis-diamminedichloroplatinum II (cisplatin)-induced nephrotoxicity. We have already demonstrated that cephaloridine, a nephrotoxic antibiotic, leads to extracellular signal-regulated protein kinase (ERK) activation in the rat kidney, which probably contributes to cephaloridine-induced renal dysfunction. The aim of this study was to examine the effect of cisplatin on ERK activation in the rat kidney and also the effect of FTS on cisplatin-induced nephrotoxicity in rats. In vitro treatment of LLC-PK1 cells with FTS significantly ameliorated cisplatin-induced cell injury. Treatment of rats with intravenous cisplatin for 3 days markedly induced renal dysfunction and increased platinum contents in the kidney cortex. An increase in pERK was detected in the nuclear fraction prepared from the rat kidney cortex from days 1 to 3 after injection of cisplatin. FTS suppressed cisplatin-induced renal dysfunction and ERK activation in the kidney. FTS did not influence any Pt contents in the kidney after cisplatin administration. FTS has been shown to enhance the in vivo expression of heat shock protein (HSP) 70 in the kidney cortex. The beneficial role of FTS against cisplatin nephrotoxicity may be mediated in part by HSP70, as suggested by its up-regulation in the kidney cortex treated with FTS alone. Our results suggest that FTS participates in protection from cisplatin-induced nephrotoxicity by suppressing ERK activation caused by cisplatin.

Involvement of activation of NADPH oxidase and extracellular signal-regulated kinase (ERK) in renal cell injury induced by zinc.

Zinc is employed as a supplement; however, zinc-related nephropathy is not generally known. In this study, we investigated zinc-induced renal cell injury using a pig kidney-derived cultured renal epithelial cell line, LLC-PK(1), with proximal kidney tubule-like features, and examined the involvement of free radicals and extracellular signal-regulated kinase (ERK) in the cell injury. The LLC-PK(1) cells showed early uptake of zinc (30 microM), and the release of lactate dehydrogenase (LDH), an index of cell injury, was observed 24 hr after uptake. Three hours after zinc exposure, generation of reactive oxygen species (ROS) was increased. An antioxidant, N, N'-diphenyl-p-phenylenediamine (DPPD), inhibited a zinc-related increase in ROS generation and zinc-induced renal cell injury. An NADPH oxidase inhibitor, diphenyleneiodonium (DPI), inhibited a zinc-related increase in ROS generation and cell injury. We investigated translocation from the cytosol fraction of the p67(phox) subunit, which is involved in the activation of NADPH oxidase, to the membrane fraction, and translocation was induced 3 hr after zinc exposure. We examined the involvement of ERK1/2 in the deterioration of zinc-induced renal cell injury, and the association between ERK1/2 and an increase in ROS generation. Six hours after zinc exposure, the activation (phosphorylation) of ERK1/2 was observed. An antioxidant, DPPD, inhibited the zinc-related activation of ERK1/2. An MAPK/ERK kinase (MEK1/2) inhibitor, U0126, almost completely inhibited zinc-related cell injury (the release of LDH), but did not influence ROS generation. These results suggest that early intracellular uptake of zinc by LLC-PK(1) cells causes the activation of NADPH oxidase, and that ROS generation by the activation of the enzyme leads to the deterioration of renal cell injury via the activation of ERK1/2.

Percutaneous sonographically guided radiofrequency ablation with artificial pleural effusion for hepatocellular carcinoma located under the diaphragm.

OBJECTIVE: Sonographically guided radiofrequency ablation is usually one of the most effective treatments for hepatocellular carcinoma. However, the localization of the tumor is a major limiting factor in the use of a sonographically guided procedure. In our experience, sonographic examination with artificial pleural effusion has improved the visualization of hepatocellular carcinomas under the diaphragm. We investigated the safety, benefits, and local efficacy of radiofrequency ablation with artificial pleural effusion. SUBJECTS AND METHODS: Twenty-five lesions in 23 patients were treated using radiofrequency ablation with artificial pleural effusion, for which 5% glucose solution was injected into the pleural cavity. RESULTS: Artificial pleural effusion allowed us to visualize the whole tumor on gray-scale sonography in 22 lesions that were not detectable or were poorly visible and to obtain a safer and easier puncture line in 14 lesions. In 23 (92%) of the 25 lesions, artificial pleural effusion was helpful in performing percutaneous radiofrequency ablation. Complete necrosis after radiofrequency ablation was obtained in 22 (88%) of the 25 lesions. During a mean (+/- SD) follow-up period of 10.6 +/- 6.5 months, local recurrence at the ablation site was diagnosed in only one (4.5%) of the 22 lesions. Mild cough in three patients and mild dyspnea in two patients were observed as adverse effects of artificial pleural effusion, but these effects were temporary. Oxygen saturation of the blood during artificial pleural effusion was slightly decreased. CONCLUSION: Radiofrequency ablation with artificial pleural effusion is a safe and beneficial treatment option that offers excellent local control through visualization of hepato-cellular carcinomas under the diaphragm.

The influence on liver parenchymal function and complications of radiofrequency ablation or the combination with transcatheter arterial embolization for hepatocellular carcinoma.

Objectives: We investigated how radiofrequency ablation (RFA) alone or the combination with transcatheter arterial embolization (TAE-RFA) for hepatocellular carcinoma (HCC) affects the liver parenchymal function of underlying chronic liver disease to find a predictive factor for selecting appropriate candidates for RFA. Methods: In 53 HCC patients (RFA alone, 25; TAE-RFA, 28), liver laboratory tests and development of complications were monitored until 6 months after treatments. Serum albumin deteriorated within 1 month and recovered after 6 months in both groups, whereas the elevation of the Pugh score continued in RFA alone group and that of serum bilirubin continued in TAE-RFA group until 6 months. The elevation of Pugh score 6 months after RFA was significantly more frequent in patients with a high pre-treatment Pugh score (>/=8 points) than in those with a low pre-treatment Pugh score (<8 points). Complications after RFA were observed in 11 of the 53 patients: temporary complications in 5, refractory ascites in 6. Refractory ascites developed in only patients with a high pre-treatment Pugh score. Conclusion: Liver parenchymal functions in patients with a low pre-treatment Pugh score were transiently deteriorated by RFA or TAE-RFA. However, in patients with a high Pugh score, RFA or TAE-RFA induces long-term deterioration of the liver parenchymal functions and causes serious complications. Therefore, patients with a Pugh score >/=8 points would not be good candidates for RFA or TAE-RFA.

Qualitative assessment of tumor vascularity in hepatocellular carcinoma by contrast-enhanced coded ultrasound: comparison with arterial phase of dynamic CT and conventional color/power Doppler ultrasound.

The purpose of this study was to evaluate the detection rate of tumor vessels and vascularity in hepatocellular carcinoma (HCC) by contrast-enhanced coded US using Levovist, and to compare with conventional color/power Doppler US (CDUS) and dynamic CT. Ninety nodules (72 hypo/isoechoic nodules, 18 hyperechoic nodules) in 61 patients were studied. We observed tumor vessels by continuous transmission at the early vascular phase (40 s following administration of Levovist) and vascularity by intermittent transmission (intervals of 2-3 s) at the late vascular phase (40 to approximately 120 s). The detection rate of tumor vessels at the early vascular phase was 97% in hypo/isoechoic nodules and 70% in hyperechoic nodules with high density in dynamic CT being higher than that by CDUS. Tumor vascularity at the late vascular phase in hypo/isoechoic and hyperechoic nodules was hyper-enhancement in 78 and 40%, iso-enhancement in 19 and 40%, and hypo-enhancement in 3 and 0%, respectively. The detection rates of tumor vessels and vascularity in hyperechoic nodules were similar to those by CDUS. The detection rates of tumor vessels and vascularity were not affected by the tumor size in HCC tumors with high density in dynamic CT. Contrast-enhanced US with Levovist was superior to CDUS and equal to dynamic CT to assess tumor vessels in hypo/isoechoic nodules. Although it was equal to CDUS for hyperechoic nodules, this modality is useful in evaluating tumor hemodynamics.

Expression of matrix metalloproiteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in hepatocellular carcinoma tissue, compared with the surrounding non-tumor tissue.

Matrix metalloproteinases (MMPs) have been implicated in the tumor invasion and growth through the degradation of extracellular matrix. In this study, we selected 46 hepatocellular carcinoma (HCC) cases, at random, and we immunohistologically examined the expression of MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, tissue inhibitor of metalloproteinases (TIMP)-1, TIMP-2, in cancerous and non-cancerous areas using avidin-biotin-peroxidase complex method. In all cases, cancer cells, hepatocytes, sinusoidal lining cells, leukocytes, and bile ducts were positive for all the primary antibodies. The expressions of MMPs and TIMPs in most of the HCC tissues were equal or low compared with those in the surrounding non-tumor tissues, although mixed expression pattern were recognized in some HCC tissues. The difference of MMP and TIMP expression was not related with the histological differentiation of HCC and the condition of non-cancerous area. These findings suggested little association of the clinicopathological findings of HCC with the histological expression of MMPs and TIMPs.

Hepatocellular carcinoma with sarcomatous change arising after radiofrequency ablation for well-differentiated hepatocellular carcinoma.

We report a case involving well-differentiated hepatocellular carcinoma (HCC) developing to HCC with sarcomatous changes after radiofrequency ablation (RFA). In a cirrhotic patient with both hepatitis B surface antigen and hepatitis C virus RNA, a well-differentiated HCC with a diameter of 2 cm was detected in segment IV of the liver. A combination of transcatheter arterial embolization and percutaneous ethanol injection (PEI) was performed and, after 8 months, PEI was performed for recurrent tumors. Fifteen months after the first treatment, a recurrent tumor measuring 3.5 cm in diameter was detected in segment IV, which was demonstrated as well-differentiated HCC by tumor biopsy, and treated by RFA. Although the treated lesion was reduced to 2.5 cm in diameter 6 months after RFA, the tumor rapidly enlarged to 6 cm in diameter 2 months later and progressed to lymph node metastasis. Aspiration biopsy showed spindle-shaped sarcomatoid cells with positive staining for both vimentin and keratin. The patient died of HCC progression 10 months after RFA. Autopsy findings showed both sarcomatoid cells and trabecular HCC cells. The sarcomatoid cells had metastasized to the lymph nodes and distant organs. This is the first case illustrating a sarcomatous HCC after RFA. Of interest, RFA may be related to the development of sarcomatous HCC.