Evaluation of distribution patterns for copper and zinc in metallothionein and superoxide dismutase in chronic liver diseases and hepatocellular carcinoma using high-performance liquid chromatography (HPLC).

It has been reported that the copper (Cu) content of hepatocytes increases in chronic liver diseases and small hepatocellular carcinoma (HCC). In cells, Cu exists mainly as Cu-metallothionein (MT) or Cu, zinc (Zn)-superoxide dismutase (SOD). In this study, we investigated the biochemical state of Cu in the hepatocytes of patients with HCC using high-performance liquid chromatography (HPLC). The subjects of present study were 23 patients with HCC who underwent liver resection. The cancerous tissue and non-cancerous hepatic parenchyma with chronic disease were analyzed. In addition, as a normal control, hepatic tissue was collected at autopsy from 13 patients with no liver disease. Each sample was diluted with buffer, chilled, homogenized, and centrifuged. The supernatant was fractionated using HPLC. The metal contents of each fraction were measured using a desktop-type inductively coupled plasma (ICP) emission spectrochemical analyzer. HPLC analysis showed that MT existed mainly as Zn-MT in the normal hepatic tissue. The case of Cu,Zn-MT was significantly greater than Zn-MT in the non-cancerous, but diseased hepatic parenchyma than in the normal hepatic tissue (p<0.01). In comparison with non-cancerous hepatic parenchyma, the Cu-MT in the cancerous section was significantly greater than the Cu,Zn-MT (p<0.01). The Cu content for MT was significantly higher in small HCC (<40 mm) (p<0.01), and the absence of Cu or Zn in the MT fraction was significantly more frequent in the large HCC (>or=40 mm) (p<0.01). The Cu and Zn content for SOD in the samples showed no significant difference. Increase in the Cu content in the cancerous hepatic tissue were, thought to be reflecting changes in the distribution of Cu in the MT fraction of hepatic tissues.

Metal contents of liver parenchyma after percutaneous ethanol injection or radiofrequency ablation in patients with hepatocellular carcinoma before and after trientine hydrochloride therapy.

We administered trientine hydrochloride, a drug used in the treatment of Wilson's disease, to patients with hepatocellular carcinoma after radical treatment with percutaneous ethanol injection or radiofrequency ablation, and examined its effect on the reduction of liver-tissue copper content. We enrolled 24 patients with 3 or fewer primary lesions of Child class A or B hepatocellular carcinoma with diameters of 3 cm or less who had undergone radical treatment with percutaneous ethanol injection or radiofrequency ablation. Trientine hydrochloride was orally administered in a single daily dose of 250 mg to 12 patients before a meal (at fasting, group 1) or at a total daily dosage of 750 mg, divided into 3 doses, to 12 patients (group 2). This study was a randomized between-groups comparative study of 12 weeks' duration. We used the particle-induced x-ray-emission method to determine liver-tissue mineral content. Urine copper and serum mineral levels were also measured, and transaminase levels were examined. Liver-tissue copper content decreased significantly, to 160.1 microg/g dry weight, after treatment, compared with the pretreatment level of 306.8 microg/g dry weight (P <.05). We detected no significant difference in iron or zinc content before and after treatment. The copper content was significantly reduced after treatment in both groups (P <.05). The urine copper level was significantly increased after 1 week of treatment but decreased thereafter. Serum copper levels were significantly reduced after treatment (P <.01). We detected no significant difference in transaminase level before and after treatment. Iron-deficiency anemia in 1 patient after 12 weeks' treatment was the only adverse reaction, and it was improved by the administration of an iron product. We noted no other overt adverse reactions. In patients with hepatocellular carcinoma, trientine hydrochloride therapy may significantly reduce copper content in liver tissue.

Radioprotective effect of chitosan in sub-lethally X-ray irradiated mice.

The radioprotective effect of chitosan was studied in mice following whole-body X-ray irradiation. C3H/He mice were exposed to 7 Gy, and their survival rates were examined. The survival rates of chitosan-diet mice were about 20% higher than those of mice on a standard diet, and the rates dropped sharply to a plateau at day 10 after X-ray irradiation. The chitosan-diet mice had an increased weight ratio of spleen to body within the experimental period. The leukocyte, thrombocyte, and erythrocyte counts as well as the hematocrit and hemoglobin levels were recovered significantly and more rapidly in the chitosan-diet mice than the standard-diet mice at day 14 after irradiation. The scavenging abilities of chitosan were evaluated by the ESR spin-trapping method. These observations suggested that chitosan led to hematopoetic activation and leukocytogenesis in mice after sub-lethal dose irradiation, and that the biological response might be caused by radical trapping or scavenging.

Metal contents in the liver of patients with chronic liver disease caused by hepatitis C virus. Reference to hepatocellular carcinoma.

BACKGROUND/AIMS: The aim of this study was to investigate the relationship between trace metals and the prevalence of hepatocellular carcinoma (HCC) in patients with chronic hepatitis or hepatic cirrhosis caused by hepatitis C virus (HCV). METHODS: We measured the contents of copper, iron, and zinc in HCC tissue (n = 112), dysplastic nodules (n = 7), and liver parenchyma in patients with (n = 112) and without (n = 12; 7 with grade F3 fibrosis, 5 with grade F4 fibrosis) HCC. Metals were quantified in thin-needle biopsy specimens using the particle-induced X-ray emission method (PIXE). RESULTS: Copper level in liver parenchyma was higher in patients with HCC than in those without HCC (p < 0.01), while there was no such difference in hepatic iron. In patients with grade F4 fibrosis, copper content in the liver parenchyma was higher in the presence of HCC than in its absence (p < 0.05). Multiple regression analysis showed that the only factor significantly associated with the coexistence of HCC in HCV-positive patients with chronic liver disease was the copper level in the liver parenchyma. CONCLUSIONS: Hepatic copper overload may contribute to the development of HCC in HCV-positive patients with chronic hepatitis or cirrhosis.

Profile of trace element concentrations in the feto-placental unit in relation to fetal growth.

BACKGROUND: During pregnancy, trace elements are indispensable for life maintenance not only for the mother but also for the fetus. The purpose of this study was to examine whether fetal growth is associated with altered levels of trace elements in maternal blood, fetal blood, and placenta tissue. METHODS: Twenty-one pairs of healthy mothers and their newborns with intrauterine growth restriction delivered after 34 weeks of gestation were recruited for the study. In addition, 30 pairs of healthy mothers and their appropriate for gestational age newborns were included as controls. Maternal venous blood, umbilical cord venous and arterial blood, and placenta tissue were collected immediately after delivery. Six essential elements, magnesium, manganese, iron, copper, zinc, and selenium, and four other elements, rubidium, strontium, cadmium, and cesium, in those samples were determined by inductively coupled plasma mass spectrometry or inductively coupled plasma atomic emission spectrometry. RESULTS: Compared with appropriate for gestational age cases, intrauterine growth restriction cases showed higher magnesium, copper, and selenium concentrations in umbilical cord arterial sera, and higher magnesium and selenium concentrations in placenta tissue, but no significant differences appeared for the elements measured in maternal and umbilical cord venous sera. The umbilical cord venous vs. maternal sera concentration ratio was elevated for copper, and the umbilical cord arterial vs. umbilical cord venous sera concentration ratios were elevated for copper and zinc, but there were no differences in placenta tissue vs. maternal sera concentration ratios in intrauterine growth restriction cases. CONCLUSIONS: Among the trace elements evaluated, magnesium, copper, zinc, and selenium showed elevated concentrations in umbilical cord arterial blood, or elevated umbilical cord arterial vs. umbilical cord venous blood concentration ratios in intrauterine growth restriction cases. Reduced consumption efficiency of these four essential trace elements may be closely associated with retarded fetal development.

Reconstruction of two victims' posturing based on the induced radioactivities in their bones in the criticality accident in Tokai-Mura, Japan.

On 30 September 1999, three workers were severely exposed to neutrons and gamma rays in a criticality accident that occurred at a uranium conversion facility in Tokai-mura, Ibaraki Prefecture, Japan. Radiochemical analyses of 32P and 45Ca induced by neutrons in bone matrix were carried out after the deaths of two of the victims. It was found that more than several million becquerels of both nuclides had been produced in their body skeletons. Results showed non-homogeneous distributions of neutron fluence in the bodies, from which it could be deduced how both workers were positioned relative to the fission source during exposure, i.e., at the moment of the first nuclear excursion. For the victim who died first, the activities in the central part of his body were more than those of his extremities. Also, in the central part of his body, the right side showed more activities than the left side. As for the second man, the activities indicated rather uniform exposure to neutrons to the whole body although the geometrical distribution of the activity varied enough to assume his orientation. Such information on the geometrical distribution of neutron-induced radioactivities in the skeleton can be used to reconstruct the posturing of the victims, which is necessary to estimate their apparent absorbed doses.

Determination of 32P in urine for early estimation of the neutron exposure level for three victims of the JCO criticality accident.

In the criticality accident which occurred on 30 September 1999 at a uranium conversion facility in Tokai-mura, Japan, three workers were severely exposed to neutron and gamma-ray irradiation. Preliminary estimations of doses from blood properties and 24Na concentration in blood were 16-20, 6-10 and 1-4 gamma-ray gray-equivalent (gammaGyEq) respectively for the three workers. For apparent dose estimation, neutron-induced radionuclides in biological materials such as blood, hair and urine were measured. Accordingly, we detected 32p in urine samples. The concentration ratios of 32P in the urine for the three workers showed a similar tendency to those of 24Na in blood. This result indicated that the radioactivity of 32P in urine could be used to estimate the neutron exposure level.

Small hepatocellular carcinoma: assessment with T1-weighted spin-echo magnetic resonance imaging with and without fat suppression.

OBJECTIVE: We examined the detectability of small hepatocellular carcinomas (HCCs) and the factors that affect hyperintensity of small HCC on T1-weighted images (T1W) by using T1-weighted fat-suppressed images (T1FS). METHODS: Thirty-nine HCCs (29 patients) measuring 30 mm or less were enrolled. The mean size of HCCs was 21.0+/-4.9 mm. Spin-echo T1W, T2-weighted images (T2W), and T1FS were obtained using a 1.5 T system. We evaluated the detectability in each sequence by receiver-operating-characteristic (ROC) analysis and the tumor-to-hepatic parenchyma contrast-to-noise ratio (CNR), the variance in the detectability among all interpreters with each sequence, and the presence or absence of improvement in the detectability by interpreting T1FS in addition to conventional T1W plus T2W. The contents of fat, copper, and iron in histologically diagnosed HCCs showing hyperintensity on both T1W and T1FS were measured. For determination of heavy metals, we used a particle induced X-ray emission analytical instrument. RESULTS: ROC analyses revealed that T1FS were superior to T1W and T2W in detecting small HCCs (0.900+/-0.017 for T1FS, 0.859+/-0.019 for T1W, and 0.745+/-0.030 for T2W). The detectability by interpreting T1FS in addition to conventional T1W plus T2W was improved (0.931+/-0.013 for the conventional images and 0.973+/-0.008 for the conventional images plus T1FS, P<0.001). The detected lesions on T1FS demonstrated favorable CNR values. The copper content in the cancer and the ratio of the copper content in the cancer to that in the non-cancerous tissue were 275.4+/-219.0 microg/g dry weight, 6.9+/-5.5 in HCCs showing hyperintensity on both T1W and T1FS. Both were significantly higher (P<0.05). CONCLUSION: T1FS showed excellent sensitivity and specificity in detecting small HCCS irrespective of the experience of interpreters. The use of T1FS suggested the involvement of copper might be one of the factors in hyperintensity of HCCs on T1W.