Alcohol consumption appears to protect against non-alcoholic fatty liver disease.

BACKGROUND: Moderate alcohol consumption may have certain beneficial effects against non-alcoholic fatty liver disease, which is associated with metabolic syndrome. AIM: To determine the association between drinking pattern and fatty liver in Japanese men and women. METHODS: A cross-sectional study was performed with health checkup data including information concerning alcohol consumption and ultrasonographic assessment of fatty liver. RESULTS: We analysed 4957 men and 2155 women without reported liver diseases (median age, 49 years). In men, 40% of nondrinkers and 28% of drinkers had fatty liver. Alcohol consumption was inversely associated with fatty liver (adjusted odds ratio, 0.54; 95% confidence interval, 0.46-0.63). The prevalence of fatty liver in each category of drinking frequency was 38% (1-3 days/week), 29% (4-6 days/week), and 24% (daily drinking); there was a significant inverse correlation between drinking frequency and the prevalence of fatty liver (P < 0.001). In women, 16% of nondrinkers and 10% of drinkers had fatty liver. Drinking less than 20 g on 1-3 days/week was associated with low prevalence of fatty liver (adjusted odds ratio, 0.47; 95% confidence interval, 0.23-0.96). CONCLUSIONS: Alcohol consumption appears to protect against non-alcoholic fatty liver disease.

Expression of alpha-fetoprotein and albumin genes in human hepatocellular carcinomas: limitations in the application of the genes for targeting human hepatocellular carcinoma in gene therapy.

For an approach of gene therapy for hepatocellular carcinoma (HCC), transcriptional regulatory sequence (TRS) of either alpha-fetoprotein (AFP) or albumin has been used for targeting cancer cells. To examine the feasibility of using TRSs of these genes for possible gene therapy of HCCs, the cellular distribution of AFP and albumin gene transcripts was studied in 25 cases of surgically removed human HCCs. AFP gene expression was observed in HCC nodules of 13 cases. The expression in HCC was heterogeneous, and the distribution of the transcripts was mostly sparse and spotty. The higher the serum AFP levels, the larger population of the AFP-expressing HCC cells tended to reflect. In noncancerous liver, a slight AFP expression was found by Northern blot analysis, but the transcripts were not detected in the liver sections. In contrast, albumin expression was found in all HCCs as well as in noncancerous hepatocytes. In HCC, the transcripts for albumin were distributed in cancer cells, and the expression varied with nodules. There were more albumin-expressing cancer cells than the AFP-expressing cells. Albumin expression was retained even in poorly differentiated HCC, although the intensity of the signal was not as strong as in more-differentiated HCCs. Metastatic HCC nodules revealed transcripts for both AFP and albumin genes, and those were clearly recognized in the lung tissue. These results suggest that, for gene therapy for HCCs, neither AFP nor albumin are ideal options for targeting HCC cells. AFP-TRS may be used as a transcriptional regulator in selected cases in which AFP gene expression is observed in the cancer nodules. The serum AFP level appears to be an important indicator in selecting cases. Albumin-TRS in conjunction with retroviral vector might be used in limited cases such as HCCs with no AFP expression. However, careful consideration must be taken, because albumin is constitutively expressed in normal hepatocytes, and AFP-expressing nonmalignant progenitor cells possibly exist.

Proliferating cell nuclear antigen and grade of malignancy in small hepatocellular carcinoma--evaluation in US-guided specimens.

BACKGROUND/AIMS: We investigated the value of the proliferating cell nuclear antigen labeling index ratio (PCNA-LI ratio: PCNA-LI of cancer/PCNA-LI of surrounding non-cancerous liver tissue) to clarify the prognosis of the patients bearing small hepatocellular carcinomas (HCC) less than 20 mm in diameter and treated by percutaneous ethanol injection therapy (PEIT). MATERIAL AND METHODS: Twenty eight HCC patients who had received PEIT were divided into 3 groups. The non-recurrence (NR) group in which no new lesions were observed for at least 18 months after PEIT (13 patients), the early recurrence group (ER) in which lesions recurred within one year (6 patients), and the late recurrence group (LR) in which lesions recurred more than one year after PEIT (9 patients). Immunohistochemical staining of PCNA was done using biopsied specimens. RESULTS: The PCNA-LI ratio in 37 well differentiated, 13 moderately differentiated, 11 poorly differentiated HCC were 1.86 +/- 0.55, 3.33 +/- 0.51, and 4.75 +/- 0.81 (mean +/- SD), respectively. The ratio in ER group (4.57 +/- 0.57) was significantly higher than that in LR (2.04 +/- 0.61) and NR group (1.87 +/- 0.62) and the PCNA-LI ratio tended to correlate with the periods until the development of recurrent lesions in cases of ER group. CONCLUSIONS: These results indicate the PCNA-LI ratio, in conjunction with the histological grade, is a useful marker for evaluating the grade of malignancy, and for predicting the period until recurrence after treatment of small HCC.

Cellular distribution of 92-kd type IV collagenase/gelatinase B in human hepatocellular carcinoma.

To examine the possible involvement of gelatinase B in human hepatocellular carcinoma (HCC), cellular localization of transcripts and protein of gelatinase B were studied by using in situ hybridization and immunohistochemistry. Transcripts for gelatinase B were observed in tumor cells in 22 cases of 27 HCCs and also in dysplastic nodules. However, there was no significant difference in the expression among histological grades of HCC. The expression was mostly homogeneous, but the intensity varied with the nodules. Of 13 cases with capsular invasion, 12 expressed gelatinase B, whereas 10 of 14 without capsular invasion did (P < 0.05). Gelatinase B transcripts were commonly observed in the sinusoidal cells of the hepatic lobules, in mesenchymal cells both in fibrous capsules and around the necrosis, and also in some undefined cells of the portal tracts of noncancerous liver. Localization of gelatinase B protein was mostly similar to but sometimes different from that of the transcripts in cancer nodules. In conclusion, the expression of gelatinase B appears to be an important characteristic of malignant transformation of hepatocytes. The findings suggest that gelatinase B synthesized by cancer cells plays an important role in the growth and invasion of HCC by degrading surrounding extracellular matrices.

Usefulness of indocyanine green injection during ultrasound-guided liver biopsy for the diagnosis of small hepatocellular carcinoma.

To diagnose hepatocellular carcinoma (HCC) functionally and immediately, we examined the usefulness of indocyanine green (ICG) injection during ultrasound-guided liver biopsy. Liver specimens were obtained after intravenous ICG injection by ultrasound-guided biopsy from 251 space-occupying lesions (SOL) in 136 patients. The tissues were immediately examined for ICG uptake using an infrared Vidicon camera and were also subjected to histopathological examinations. Of the 112 ICG-negative biopsy specimens, 105 were histologically diagnosed as HCC, 6 as dysplastic nodules (DN) and 1 as a regenerative nodule (RN). Of the 139 ICG-positive specimens, 18 were diagnosed as HCC, 1 as DN and 120 as RN. The sensitivity of the absence of ICG uptake (SEAIU), the specificity of the absence of ICG uptake (SPAIU), and the positive predictive value of the absence of ICG uptake (PPAIU) for the diagnosis of HCC were 85.3%, 94.5% and 93.8%, respectively. Of the 251 SOLs, 184 were less than 2 cm. SEAIU, SPAIU and PPAIU for the diagnosis of these small HCC were 85.3%, 94.5% and 91.4%, respectively. These results support the reliability of ICG injection during ultrasound-guided liver biopsy to diagnose even small HCC.

Cellular distribution of transcripts for tissue inhibitor of metalloproteinases 1 and 2 in human hepatocellular carcinomas.

The cellular distribution of tissue inhibitor of metalloproteinases (TIMP)-1, and TIMP-2 was studied by using in situ hybridization in surgically removed human hepatocellular carcinomas (HCCs) and cholangiocellular carcinomas (CCCs). The purpose of this study was to characterize the protein involvement of TIMPs in the development of HCCs and CCCs. All HCCs and CCCs expressed TIMPs. The distribution of transcripts for TIMPs in the tumors was mostly homogeneous. Expression of TIMP in cancer cells was more intense than that in the surrounding noncancerous liver (either, cirrhosis, chronic hepatitis, or normal), and expression of TIMP-1 was stronger than that of TIMP-2. Expression of TIMPs varied among HCC nodules, but there was no obvious association between the expression level of TIMPs and differentiation stages or invasiveness of the HCCs. Transcripts for TIMPs were clearly demonstrated in the metastatic HCC nodules in the lung. Expression of TIMP-1 CCC was strong, and small nodules of CCC were recognized in the liver. Immunohistochemical study for TIMP-1 revealed a consistent staining of the TIMP protein with the transcripts. In the peritumoral histologically normal liver, which was not infected with either hepatitis B or C virus, expression of TIMP-1 was found in various cell types, but that of TIMP-2 was weak. Expression of TIMP-1 in hepatocytes revealed clear zonal distribution. These results suggest that TIMPs may act on modulating the matrix/tumor interaction and may play an important role in growth and invasion of HCCs and CCCs. Expression of TIMP-1 can be a marker of HCC metastasis to the lung, and also that of the extent of CCC invasion. Furthermore, the consistent expression of TIMPs in many cell types of the noncancerous liver suggests some unknown functional role that must be clarified.

Copper-induced hypercholesterolemia of golden hamsters: enhanced synthesis of cholesterol in the liver.

Effects of a subtoxic dose of copper on cholesterol metabolism were studied in male golden hamsters. Intraperitoneal injections of cupric acetate increased serum levels of cholesterol and phospholipids without liver damage. This lipidemia was associated with increased cholesterol of the liver. The participation of hemolysis was denied by peripheral red blood cell tests. Hepatic microsomal 3-hydroxy-3-methyl-glutaryl-CoA reductase was also elevated by copper administration. Biliary secretion of cholesterol increased but that of bile acids remained unchanged, suggesting no impaired degradation of cholesterol. We conclude that hepatic synthesis of cholesterol is enhanced by a subtoxic dose of copper, resulting in hypercholesterolemia.

Analyses on lipid-rich organelles of the rat liver after bile duct ligation.

A fraction of lipid-rich organelles of rat livers was analyzed to study the effect of bile duct ligation on lipid metabolism. The lipid fraction of the control rats consisted mainly of Golgi-derived dense bodies. Three days after the ligation, myelin-like figures and increased phospholipids and cholesterol were characteristic of the fraction. Two weeks ligation resulted in proliferation of lysosomal electron dense bodies. As their volume density expanded, phospholipids and cholesterol increased. Electrophoresis indicated a reduced rate of VLDL assembly after ligation. The Golgi-dominant phase is converted to the lysosomal phase in the heavy subfraction after the bile duct ligation. This subcellular change could be a consequence of secondary lipidosis: hepatocyte lysosomes have been exhausted in sorting accumulated lipids into secretory lipoproteins.

A copper-sulfur complex in the liver of a patient with Wilson's disease.

An asymptomatic 16-year-old boy was found to have Wilson's disease without Kayser-Fleischer rings. Liver biopsy showed chronic active hepatitis with 1025 micrograms copper/g dry weight. After 19 months of d-penicillamine therapy, the liver histology became almost normal and the copper content decreased to 238 micrograms/g dry weight. The liver specimens obtained before and after treatment were studied by X-ray probe microanalysis. After treatment, both copper and sulfur decreased in hepatocellular lysosomes. The estimated molar ratio of the decreased copper to the decreased sulfur was 32/100. These figures suggest that lysosomal copper exists in the form of metallothionein.

An association of mercury with selenium in inorganic mercury intoxication.

Energy dispersive X-ray analysis was performed on the renal tubular cells of two patients with inorganic mercury intoxication. Some lysosomes of these cells consisted of unusual matrices of aggregated electron-dense grains which were positive for mercury, selenium and sulphur. Though maps of the specific X-rays of both mercury and selenium coincided exactly with these lysosomes, the molecular ratio of selenium to mercury ranged between zero and 2.9. It is unlikely that the trace element of selenium and exogenous inorganic mercury are deposited in the lysosomes independent of each other, but rather their coexistence in the characteristic lysosomes strongly suggests a compound formed by binding mercury to the SeH residues of selenoprotein.

Amosulalol, a combined alpha and beta adrenoceptor antagonist: kinetics after intravenous and oral doses.

Amosulalol kinetic studies were conducted in seven subjects who received 0.16 mg/kg iv and in 18 subjects who received 12.5, 25, 50, 100, or 150 mg by mouth. Plasma levels of amosulalol after intravenous dosing declined biphasically and fitted a two-compartment model. Kinetics were as follows: coefficients A = 0.85 +/- 0.09 microgram/ml and B = 0.22 +/- 0.01 microgram/ml; rate constants alpha = 2.78 +/- 0.24 hr-1 and beta = 0.25 +/- 0.01 hr-1; elimination rate constants, k12 = 1.36 +/- 0.17 hr-1, k21 = 0.78 +/- 0.06 hr-1, and kel = 0.88 +/- 0.06 hr-1; terminal phase volume of distribution = 0.75 +/- 0.06 l/kg; clearance = 8.09 +/- 0.54 l/hr; AUC = 1.22 +/- 0.09 microgram . hr/ml; and t1/2 alpha = 0.26 +/- 0.02 hr and t1/2 beta = 2.8 +/- 0.1 hr. After single oral doses, amosulalol peak plasma levels were generally reached within 2 to 4 hr. Maximum plasma concentrations and AUC increased in a dose-dependent manner, whereas t1/2 were about 5 hr (range 4.4 to 5.7 hr) at each dose. Systemic availability of amosulalol was about 100% as determined by the ratio of AUC after oral and intravenous dosing. These results suggest that amosulalol is well absorbed and is little affected by first-pass metabolism.

Changes in the effects of clonidine on left atrium and hindlimb vasculature of rats in various thyroid states. A study of the responsiveness of alpha 2-adrenoceptors in the cardiovascular system.

Postsynaptic alpha 2-adrenoceptors have been reported to exist in various tissues, including vascular smooth muscle. In order to investigate the possibility of their mediating a positive inotropic change and clarify the influence of the thyroid on their responsiveness, we examined the effects of clonidine, a known alpha 2-agonist, on the isolated left atria and femoral vascular beds of rats which were made hypo-, hyper- or euthyroid. Clonidine caused a dose-dependent positive inotropic change in the hypothyroid rat atrium, which was thought to be due to its alpha 1-stimulating action because of the antagonistic effect exerted by either phentolamine (10(-6) M) or prazosin (10(-7) M), but not by yohimbine (10(-7) M) or cimetidine (10(-5) M). In the hyperthyroid rat atrium, clonidine exerted a negative inotropic effect at high concentrations, which was thought to be due to its direct action on the cardiac muscle. Clonidine did not cause any inotropic change in the euthyroid rat atrium. Thus, an inotropic change mediated by the postsynaptic alpha 2-adrenoceptors could not be demonstrated in the rat heart. In the experiment involving hindlimb perfusion, clonidine caused vasoconstriction which was antagonized by yohimbine (10 micrograms/min). This effect was significantly augmented in the hypothyroid rats but not changed in the hyperthyroid ones. The vasoconstrictive effect of phenylephrine was found to be reduced in both hypo- and hyperthyroid rats. These results suggest that, in the peripheral vascular system, thyroid function also influences the postsynaptic alpha 2-adrenoceptors, but not in the same way as it affects the alpha 1-adrenoceptors.

Induction of opposite response of hindlimb vasculature to adrenergic agents in hypothyroid rats.

In order to investigate whether hypothyroidism induces some functional change in the vascular smooth muscle, the responsiveness of the femoral vascular bed of hypothyroid rats to vasoactive agents was compared with that of euthyroid rats. The vascular beds were perfused by using the constant flow method. Ad libitum feeding of 0.15% 6-propylthiouracil to young male Wistar rats (120 g) over 3 months leads to hypothyroidism. Hypothyroidism did not induce any changes in the Ach- or papaverine-induced vasodilation and 5-HT-induced vasoconstriction. In contrast to euthyroidism, hypothyroidism induces a marked augmentation of the clonidine-induced vasoconstriction, while the phenylephrine-induced vasoconstriction and isoprenaline-induced vasodilation were clearly attenuated. The response to phenylephrine in euthyroidism was predominantly inhibited by prazosin and was slightly modified by yohimbine. While the attenuated response to phenylephrine in hypothyroidism was modified by these blockers in a converse manner, the response to clonidine in each rat group was not modified by prazosin, but markedly inhibited by yohimbine. From these results the following conclusions were obtained: both alpha-1 and alpha-2 adrenoceptors in the femoral vascular beds of eu- and hypothyroid rats responded to phenylephrine; phenylephrine stimulated predominantly alpha-1 adrenoceptors in euthyroidism; hypothyroidism caused an attenuation of the responsiveness mediated through alpha-1 adrenoceptors and an augmentation of that of alpha-2 receptors; hypothyroidism caused an opposite response, mediated through alpha- and beta-receptors, without influencing muscarinic and serotoninergic responses; augmentation of the alpha-2 response and attenuation of alpha-1 and beta-2 responses caused by hypothyroidism may be attributed to functional changes at receptor sites.