Value of the extracellular water ratio for assessment of cirrhotic patients with and without ascites.

AIMS: Ascites, which often complicates liver cirrhosis, is reported to be a factor that worsens the outcome. The aims of this study were to quantify body water compartment changes in cirrhotic patients, with and without ascites, and to elucidate the value of body water analysis for predicting the development of ascites. METHODS: A total of 109 cirrhotic patients, with and without ascites, and 65 controls were studied. Intra- and extracellular water (ECW) in the whole body and in the arm, leg and trunk were measured using the recently developed 8-electrodes multiple-frequency bioelectrical impedance analyzer. Furthermore, patients without ascites were followed to an episode of ascites or death. RESULTS: Patients with liver cirrhosis had significantly higher ECW ratios than controls. ECW ratios were increased in cirrhotic patients with moderate and severe disease. The ECW ratio of the trunk showed highly significant changes in cirrhotic patients with ascites. The ECW ratio correlated with age, serum albumin, and prothrombin time. A relative expansion of ECW and low albumin were predictive of further episodes of ascites (log-rank 6.94, P < 0.01). In multivariate analysis, the ECW ratio was independently associated with the development of ascites. CONCLUSION: Liver cirrhosis was characterized by a redistribution of body water. The ECW ratio is a reliable tool for quantification of redistribution of body water and can predict the development of ascites.

Impaired regulation of serum hepcidin during phlebotomy in patients with chronic hepatitis C.

AIM: This study was conducted to determine the clinical relevance of hepcidin, a recently identified key iron regulatory hormone, in patients with chronic hepatitis C virus (C-HCV). METHODS: Serum hepcidin levels were measured in 9 C-HCV patients by surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS), and compared to those of healthy controls. Sequential changes of hepcidin were also investigated during phlebotomy. RESULTS: Serum hepcidin and ferritin were significantly higher in C-HCV than in controls (P = 0.0002), these two variables were strongly related to each other (r = 0.658; P < 0.01), and phlebotomy significantly decreased serum hepcidin in C-HCV (P = 0.0007); all these results recollect the hepcidin response to iron signal. Hepcidin/ferritin ratio, an index of the appropriateness of hepcidin expression relative to iron overload, was significantly lower in C-HCV than in controls (0.33 +/- 0.41 vs. 0.73 +/- 0.36, P = 0.0068). This relative impairment of hepcidin expression was not reversible after phlebotomy (P = NS). CONCLUSIONS: Although the hepcidin expression responds to iron conditions in C-HCV, this response is relatively limited. This relative impairment of hepcidin expression may be relevant to disease progression, and thus correction of its regulation may be beneficial for these iron-overloaded C-HCV patients.

Iron overload is associated with hepatic oxidative damage to DNA in nonalcoholic steatohepatitis.

Several lines of evidence have suggested that oxidative stress plays an important role for the pathogenesis of nonalcoholic steatohepatitis (NASH). Therefore, by using immunohistochemical staining of liver biopsy samples, we measured hepatic 7,8-dihydro-8-oxo-2' deoxyguanosine (8-oxodG), a DNA base-modified product generated by hydroxyl radicals, of 38 NASH patients and compared with 24 simple steatosis and 10 healthy subjects. Relation of hepatic 8-oxodG with clinical, biochemical, and histologic variables and changes after iron reduction therapy (phlebotomy plus iron-restricted diet) were also examined. Hepatic 8-oxodG levels were significantly higher in NASH compared with simple steatosis (17.5 versus 2.0 8-oxodG-positive cells/10(5) microm(2); P < 0.0001). 8-oxodG was significantly related to iron overload condition, glucose-insulin metabolic abnormality, and severities of hepatic steatosis in NASH patients. Logistic regression analysis also showed that hepatic iron deposit and insulin resistance were independent variables associated with elevated hepatic 8-oxodG. After the iron reduction therapy, hepatic 8-oxodG levels were significantly decreased (from 20.7 to 13.8 positive cells/10(5) microm(2); P < 0.01) with concomitant reductions of serum transaminase levels in NASH patients. In conclusion, iron overload may play an important role in the pathogenesis of NASH by generating oxidative DNA damage and iron reduction therapy may reduce hepatocellular carcinoma incidence in patients with NASH.

Patients with chronic hepatitis C achieving a sustained virological response to peginterferon and ribavirin therapy recover from impaired hepcidin secretion.

BACKGROUND/AIMS: The aim of this study is to determine the clinical relevance of hepatic producing iron regulatory hormone-hepcidin, on iron overload in patients with chronic hepatitis C (CHC). METHODS: Serum hepcidin was measured in 73 CHC patients by surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS), and compared to those of healthy controls and anemia of inflammation patients, and analyzed their relationship to hepatic hepcidin mRNA expression levels and clinical, hematological, and histological findings. The sequential changes of hepcidin were investigated in 27 CHC patients treated with a 48 week-course of pegylated-interferon (PEG-IFN) plus ribavirin therapy. RESULTS: Serum hepcidin was positively correlated with hepatic hepcidin mRNA levels, serum ferritin and the degree of hepatic iron deposition in CHC. Serum hepcidin-to-ferritin ratios were significantly lower in HCV positive patients than in HCV negative controls in both hyper- and normal-ferritinemic conditions. This relative impairment of hepcidin production was fully reversible after successful HCV eradication by PEG-IFN plus ribavirin, concomitantly with the improvement of the iron overload condition. CONCLUSIONS: The impairment of hepatic hepcidin production occurring with chronic HCV infection may enhance iron toxicity and lead to disease progression, and modulation or supplementation of hepcidin may be beneficial for these conditions in CHC.

Hepcidin expression in the liver: relatively low level in patients with chronic hepatitis C.

Patients with chronic hepatitis C frequently have serum and hepatic iron overload, but the mechanism is unknown. Recently identified hepcidin, exclusively synthesized in the liver, is thought to be a key regulator for iron homeostasis and is induced by infection and inflammation. This study was conducted to determine the hepatic hepcidin expression levels in patients with various liver diseases. We investigated hepcidin mRNA levels of liver samples by real-time detection-polymerase chain reaction; 56 were hepatitis C virus (HCV) positive, 34 were hepatitis B virus (HBV) positive, and 42 were negative for HCV and HBV (3 cases of auto-immune hepatitis, 7 alcoholic liver disease, 13 primary biliary cirrhosis, 9 nonalcoholic fatty liver disease, and 10 normal liver). We analyzed the relation of hepcidin to clinical, hematological, histological, and etiological findings. Hepcidin expression levels were strongly correlated with serum ferritin (P < 0.0001) and the degree of iron deposit in liver tissues (P < 0.0001). Hepcidin was also correlated with hematological parameters (vs. hemoglobin, P = 0.0073; vs. serum iron, P = 0.0012; vs. transferrin saturation, P < 0.0001) and transaminase levels (P = 0.0013). The hepcidin-to-ferritin ratio was significantly lower in HCV(+) patients than in HBV(+) patients (P = 0.0129) or control subjects (P = 0.0080). In conclusion, hepcidin expression levels in chronic liver diseases were strongly correlated with either the serum ferritin concentration or degree of iron deposits in the liver. When adjusted by either serum ferritin values or hepatic iron scores, hepcidin indices were significantly lower in HCV(+) patients than in HBV(+) patients, suggesting that hepcidin may play a pivotal role in the pathogenesis of iron overload in patients with chronic hepatitis C.

Upregulation of transferrin receptor 2 and ferroportin 1 mRNA in the liver of patients with chronic hepatitis C.

BACKGROUND: Iron accumulation has been reported to be associated with progression of liver injury. The mechanism of iron accumulation in the liver is not known. In the present study, hepatic messenger RNA (mRNA) expression of transferrin receptor (TfR)1, TfR2, and ferroportin (FP)1 was measured in patients with chronic hepatitis (CH). METHODS: Eleven patients with CH-B and 43 patients with CH-C were enrolled. All patients underwent liver biopsy. Hepatic expression of TfR1, TfR2 and FP1 mRNA was analyzed using a real-time polymerase chain reaction. Total hepatic iron score (THIS) was evaluated by Prussian blue staining. RESULTS: Serum ferritin concentration is significantly higher in CH-C than in CH-B. Values of THIS of >/=5 were observed only in CH-C patients (44% of CH-C patients). The expression level of TfR2 mRNA was 10-26-fold higher than the TfR1 mRNA expression level. The TfR2 and FP1 mRNA expression was significantly higher in CH-C than in CH-B patients. Hepatic expression of TfR2 and FP1 mRNA was well correlated with THIS. CONCLUSIONS: Hepatic iron accumulation is more severe in patients with CH-C. Upregulation of hepatic iron transporters may contribute to the hepatic iron accumulation in CH-C.

Efficacy of long-term dietary restriction of total calories, fat, iron, and protein in patients with chronic hepatitis C virus.

OBJECTIVES: A diet restrictive in total calories, fat, iron, and protein intake reduces serum alanine aminotransferase levels in patients with long-term hepatitis C virus infection. However, whether long-term dietary therapy causes adverse effects such as malnutrition and anemia due to a shortage of energy intake is not clear. We evaluated the balance of energy intake and changes in physical and hematologic indices of nutrition after a long-term dietary therapy. METHODS: Twenty-two patients with long-term hepatitis C virus infection that did not respond to or who were able or unwilling to take interferon therapy were enrolled in this study. Our prescriptions included 7 mg/d or less of iron, 30 kcal. kg(-1). d(-1) of energy, 1.1 to 1.2 g. kg(-1). d(-1) of protein, and a fat energy fraction of 20%. Patients were followed for 24 mo. RESULTS: Mean body fat percentage was 24.6% at entry and was significantly reduced after the diet prescription. Mean serum ferritin decreased significantly from 376 ng/mL at entry to 141 ng/mL after 24 mo. Mean serum alanine aminotransferase levels decreased significantly from 66 to 49 IU/L. Mean levels of hemoglobin, serum albumin, and cholinesterase did not change significantly during the follow-up period. CONCLUSIONS: These results suggest that restriction of energy, fat, iron, and protein intakes is safely tolerated, so its long-term use should be recommended to patients with long-term infection with hepatitis C virus.

Different hepatitis C virus dynamics of free-virions and immune-complexes after initiation of interferon-alpha in patients with chronic hepatitis C.

BACKGROUND/AIMS: To elucidate the mechanisms of action of interferon (IFN) against hepatitis C virus (HCV), we studied the serum HCV dynamics of free-virions (FV) and immune-complexes (IC) in patients treated with IFN. METHODS: FV and IC were separated by immunoprecipitation using anti-human immunoglobulin and quantified serially using real-time detection-polymerase chain reaction. RESULTS: Initially [1st phase (0-24 h)], the FV decreased more rapidly compared to IC [exponential decay slope (EDS)=1.78+/-0.42 vs. 0.99+/-0.31 log10/day, P<0.001; half-life=5.65+/-2.02 vs. 12.5+/-2.83 h, P<0.0001], but at the 2nd phase (1-14 days), half-life of FV was significantly longer than that of IC (101+/-117 vs. 14.2+/-1.08 h, P<0.005). Regarding response to IFN, the decline slope was not significantly different at the 1st phase, but at the 2nd phase, the FV-HCV RNA decreased more slowly in non-responders than in sustained responders to IFN (EDS=0.05+/-0.02 vs. 0.34+/-0.19 log10/day, P<0.005; half-life=186+/-112 vs. 15.3+/-1.85 h, P<0.005). CONCLUSIONS: The presence of escape mutants from the neutralizing antibodies may be involved in resistance to IFN. Analyzes of FV- and IC-HCV dynamics are useful for predicting the IFN efficacy and understanding the mechanism of IFN action in chronic hepatitis patients.

Nonimmune complexed HCV RNA titer in serum as a predictor of interferon response in patients with chronic hepatitis C.

OBJECTIVE: Hepatitis C virus (HCV) has been reported to exist in the circulation of patients in various forms such as free virions, immune complexes, and nucleocapsids. To clarify the clinical significance of serum HCV titers according to the forms of virus particles, we evaluated the immune complexed (IC) and nonimmune complexed (NIC) HCV RNA titers in 77 chronic hepatitis patients treated with interferon (IFN). METHODS: IC and NIC forms in pretreatment serum were separated by immunoprecipitation using antihuman immunoglobulin antibody, and quantified by reverse transcription polymerase chain reaction. RESULTS: Serum titers of NIC HCV RNA were correlated with those of whole serum HCV RNA (r = 0.96, p < 0.01) and IC HCV RNA (r = 0.98, p < 0.01), but they were not with the aminotransferase levels, gamma-globulin concentration, and grading or staging of liver histology. Nonresponders to IFN had significantly high NIC HCV RNA titers compared with sustained responders (10(4.93 +/- 0.81) copies/ml vs 10(4.06 +/- 0.69) copies/ml, p < 0.01). It is noteworthy that the relative amount of NIC HCV RNA to whole serum HCV RNA was also significantly higher in nonresponders than in sustained responders (0.66 +/- 0.10 vs 0.50 +/- 0.11, p < 0.0001). Multivariate analysis showed that low NIC HCV RNA titer (p < 0.01) and genotype 2 (p = 0.02) were independent variables contributing to sustained response to IFN, but the whole serum HCV RNA titer was not. CONCLUSIONS: Pretreatment NIC HCV RNA titer is a more reliable predictive marker than genotype or whole serum HCV RNA of a sustained response to IFN monotherapy. This finding suggests that humoral immunity may affect the response to IFN.

The educational experiments of school health promotion for the youth in Japan: analysis of the 'sport test' over the past 34 years.

The objective of this study was to analyze independently the results of a 'sport test' that had been conducted in Japanese schools for 34 years without interruption, and to examine how physical education in Japanese schools affects health promotion in youths and what problems remain to be solved. The source of materials for the present study were the Annual Reports on the Survey of Physical Fitness and Athletic Ability for the period 1965-1998, published by the Ministry of Education, Science, Sports and Culture. Means and standard deviations of total scores from physical fitness and athletic ability tests in 11-year-olds (6th graders in elementary school), 14-year-olds (3rd graders in junior high school) and 17-year-olds (3rd graders in senior high school) were analyzed. The results of this study suggest that the trends of changes in mean score and coefficient of variation (CV) for physical fitness and athletic ability in Japanese youths are associated with the outcome of physical training through physical education in school, as well as the effect of the media--mainly television and television games. (1) The increase in mean scores and the decrease in CV observed from 1964 to the first half of the 1970s were probably due to the positive influence of the Guideline for Teaching. This guideline emphasized a systematic approach to athletic techniques, together with active practice of physical fitness training in schools, under the directives of the Boards of Education in response to the social 'Physical Fitness Campaign' policy at that time. (2) The leveling off of mean scores and CVs observed from the early half of the 1970s to the later half of the 1980s might be a result of the invasion of television into Japanese family life. (3) The decrease in mean scores and increase in CV observed since the later half of the 1980s were assumed to be caused by the negative influence of a Guideline for Teaching characterized by the key words 'Physical Education for More Pleasure' and a steep increase in the number of television games. (4) In terms of methodology, analyzing not only the mean scores but also the CV probably provided a more accurate evaluation of the outcome of health promotion in youths. In conclusion, strategies for health promotion in youths, especially for proper nurturing of physical fitness and athletic ability, should include not only delivery of physical fitness training in schools, but also continuous monitoring of multiple indicators, and ensuring proper 'Learning of the Body'. The latter should include providing learning opportunities centering on the 'body' (not the 'body' for the sake of labour or military manpower, but one's own 'body' per se); guiding young people to recognize the states of their own body and physical fitness, and to understand how they can be improved. To be effective in implementation, the need to increase the number of professional teachers in physical education should be examined. All possible opportunities should also be taken to inform youths of the negative effects of television and television games, and to encourage them to spend less time on these and more time being physically active from the moment they arise.

A study on bone stiffness and related factors in healthy secondary school girls / 体力科学

The purpose of this study was to examine the bone stiffness of healthy girls during their growth period and factors affecting on such stiffness. The subjects were 302 secondary school girls in Tokyo. Parameters examined included the body characteristics (standing height, body weight, bone stiffness measured by ultrasound, muscle thickness measured with the ultrasound B-mode system), extension power of the lower limb (containing the knee and hip joints), and a questionnaire about the daily intake of milk, kinds of meals and number of years from menophania. These examinations were carried out in June 1997.<BR>Results were summarized as follows : 1) From the observation of bone stiffness in secondary school girls, it appears that bone stiffness increases during the junior high school period. In other words, the bone stiffness of the subjects had almost reached on adult level by high school. 2) A significant positive correlation was recognized between chronological age and bone stiffness (r=0.365, p<0.05) . A positive correlation also existed between the years from menophania and bone stiffness (r=0.477, p<0.05) . These coefficients showed that the years from menophania correlate with bone stiffness more closely in comparison with chronological age. 3) In the period when short comparatively years from menophania, body characteristics, which were the index of maturity, correlated to bone stiffness. However, muscle thickness/power, which was related to exercise habit, became the major parameter correlating with bone stiffness. These results suggest that factors affecting bone stiffness should differ according to the growth period. 4) Comparison of different athletic clubs showed that the bone stiffness of volleyball players was higher than that of control. These data suggest that physical education class, which was given 3 times a week, is not enough for total body development in both quality and quantity. In other words, physical education should be better matched with total physical development, including the growth of healthy bones.