Possible factors affecting the cholesterol nucleation time in human bile: A filtration study

This study examined the hypothesis that filtration through the MW 300 kD cut-off membrane (XM-300) may influence factors affecting the cholesterol nucleation time (the appearance time of cholesterol monohydrate crystal). Differences in biliary lipids, biliary protein, mucous glycoprotein and vesicular composition were examined before and after the XM-300 filtration in control and cholesterol gallstone patients. The nucleation time in the cholesterol gallstone patients was significantly faster than that in the control patients. However, the nucleation time in the gallstone patients was significantly prolonged following the XM-300 filtration resulting in a similar value to that of the control patients. No differences in concentrations of total protein, mucous glycoprotein or lipids composition were observed. The nucleation time did not correlate with the total lipid concentration, the concentrations of biliary mucous glycoprotein or total protein. The prolonged nucleation time of gallstone bile by the XM-300 filtration was primarily ascribed to the removal of vesicles, which was confirmed by gel chromatography. It was concluded that vesicles greater than MW 300 kD are primarily responsible for the rapid nucleation time.     

Clinical application of a sensitive immunoradiometric assay for human growth hormone

A sensitive immunoradiometric assay (IRMA) for human growth hormone (GH) using monoclonal antibodies was evaluated for clinical application. The detection limit of GH in serum was 0.05 ng/ml using 0.1 ml of serum. The specificity and assay precision were satisfactory. GH levels in serum determined by the IRMA were well correlated to those determined by the sensitive enzyme immunoassay or the commercial radioimmunoassay. Most of the basal GH levels in serum from normal adult subjects were detectable; mean values in male and female were 0.21 and 0.65 ng/ml, respectively. Serum GH levels decreased to less than 1 (0.5) ng/ml after oral glucose loading in normal subjects. Postoperative acromegalic patients were classified into two groups according to their GH responses to glucose. In group 1, GH levels were suppressed less than 1 ng/ml, suggesting normal GH dynamics. On the other hand, in group 2, GH levels remained elevated above 1 ng/ml, and postinhibitory rebound rise in serum GH after dopamine infusion was observed, suggesting the presence of residual adenoma tissues. Determination by sensitive IRMA of GH responses during oral glucose loading is useful for evaluating the activity of postoperative acromegaly.     

The comparative potency of cholesterol crystallization-effector proteins in supersaturated model bile systems: Association with vesicle transformation

Various proteins which affect cholesterol crystallization are known to be present in bile, although the relative potency of their action is yet to be established. In this study, we evaluated the comparative potency of nucleating-effector proteins using a recently developed method for quantitative assessment of vesicle transformation in supersaturated model bile systems, to partially characterize mechanisms of their action. Concanavalin A-bound glycoproteins isolated from human gall-bladder bile shortened cholesterol crystallization time by 40% and increased cholesterol growth rate and final crystal mass by 161 and 19%, respectively, when compared to the control. In addition, immunoglobulins isolated from human gall-bladder bile increased cholesterol growth rate by 9%, but showed no significant effect on cholesterol crystallization time and final crystal mass. In contrast, human serum apolipoproteins A-I and B reduced cholesterol growth rate by 26 and 31% and reduced final crystal mass by 12 and 21%, but did not affect cholesterol crystallization time. Gel permeation chromatography revealed that proteins were distributed to both vesicles and bile salt micelles, but that no marked redistribution of lipids was caused by addition of these proteins. Furthermore, no significant difference in crystal structure was observed by video-enhanced contrast microscopy. These results indicate that nucleatingeffector substances tested in this study may modulate vesicular cholesterol-holding capacity, thus affecting cholesterol crystallization. Such modulation is based upon the protein-vesicle association which defines the physico-chemical metastability of vesicular cholesterol.     

Clinical evaluation of a newly established anti-HCV assay for the diagnosis of hepatitis C in Japan

To develop a more dependable method of diagnosing hepatitis C, serum anti-hepatitis C virus (HCV) was examined by using a new assay (anti-HCV second generation). The results were compared with those of either the conventional assay (anti-HCV first generation) or HCV-RNA analysis. With the first generation assay, anti-HCV was detected in 69% of post-transfusion acute hepatitis (AH), 44% of sporadic AH, 50% of needlestick exposed AH, 72% of chronic hepatitis (CH), 77% of liver cirrhosis (LC) and 86% of hepatocellular carcinoma (HCC). These results were remarkably increased by using the second generation assay (92% in post-transfusion AH, 72% in sporadic AH, 100% in needlestick exposed AH, 96% in CH, 96% in LC and 97% in HCC). Furthermore, in the early stages of AH (from 1-5 weeks after onset), anti-HCV was not detected in all 18 patients by the first generation assay, but was found in 10 of them by using the second generation assay. The failure to detect anti-HCV with the first generation assay was mainly due to a lack of the core region coding peptide (C22-3) in this assay. In the AH-resolving group, anti-HCV second generation did not disappear, but the titre tended to be lower than that in the CH-developing group. Thus, the second generation assay for anti-HCV was considered to be a more useful tool for not only the diagnosis of hepatitis C but also for determining prognosis.     

Liver biopsy bleeding time: An unpredictable event

The aim of this study was to observe the correlation between liver biopsy bleeding time, prothrombin time ratio and platelet count. The subjects were 51 consecutive patients referred for laparoscopic liver biopsy. The intervention was laparoscopy under local anaesthetic and liver biopsy with observation of post biopsy bleeding time. No correlation was found between observed liver biopsy bleeding time and platelet count or prothrombin time ratio. Thus, mild to moderate coagulopathy does not appear to be associated with prolonged bleeding following liver biopsy. Equally, normality of these coagulation studies does not indicate an absence of risk for post liver biopsy bleeding.     

Incidence of achalasia in New Zealand, 1980-84: An epidemiological study based on hospital discharges

One hundred and fifty-two patients with achalasia were first admitted to hospital in New Zealand between 1980 and 1984. The discharge of these patients was recorded by the National Health Statistics Centre which records details of all public health hospitals in New Zealand. The overall incidence was 0.95/100 000 population per year and was similar in North and South Islands and in men and women. The incidence increased with age and was 5.4/100 000 population per year in people aged over 80 years old. The disease was more common in Polynesian migrants (1.3/100 000 population per year) than other ethnic groups. Variations in incidence were seen across the Health Boards with the highest incidence of 2.9/100 000 population per year in Hawkes Bay and Tauranga.     

Nucleotide sequence of a hepatitis B virus genome of subtype adw isolated from a Philippine: Comparison with the reported three genomes of the same subtype

The complete nucleotide sequence was determined for a hepatitis B virus genome of subtype adw (pFDW294) isolated and cloned from the plasma sample of a Philippino. The genome was 3221 base-pair long with a point mutation at the 1376th nucleotide that affected the coding capacity of the P and X genes. There was a wide range of sequence divergence among pFDW294 and the reported three genomes of the same subtype (1.1–9.9%), occurring more often in the pre-S region and the S gene than in the pre-C region and the C gene.     

Methylprednisolone does not inhibit the release of growth hormone after intravenous injection of a novel growth hormone secretagogue in rats

The present study was undertaken to study the growth hormone-releasing properties and growth-promoting effect of a GH secretagogue ipamorelin (IPA) in rats given the synthetic glucocorticoid methylprednisolone (MP).In a first experiment, rats received either saline or MP (5.0 mg/kg) for 8 days. Treatment with MP significantly (P< 0.001) decreased body weight gain, but the acute response to either IPA or growth hormone releasing hormone (GHRH) in terms of plasma GH was not changed.In a second experiment, venous catheters were surgically implanted. On the next day, rats were randomly allocated to receive saline alone, MP alone (5.0 mg/kg) or MP plus IPA in doses of 0.4 or 1.6 mg/kg/day for 10 days. IPA was administered intravenously four times a day.MP treatment significantly (P< 0.05) retarded recovery from surgery in terms of body weight. Thus, saline treated animals lost 4.0 +/- 3.5 g over the entire experimental period, whereas animals receiving MP lost 13. 6 +/- 2.9 g. When IPA was given together with MP, losses in body weight were significantly (P< 0.05) reduced to 2.3 +/- 2.0 and 1.6 +/- 2.0 g in animals given the high and low dose of IPA, respectively. In parallel with this IGF-I levels increased.In conclusion, this work shows that MP does not disrupt the response of the GH-IGF-I axis to an exogenous stimulus like IPA, and repeated stimulation leads to increases in IGF-I and of body weight gain.     

A novel mutation at the donor splice site of intron 3 of the GH-I gene in a patient with isolated growth hormone deficiency

A G - C transversion at the fifth nucleotide of intron 3 of GH-I gene was identified in a sporadic case of isolated growth hormone deficiency (IGHD). The mutation was absent in both of the parents, indicating that the mutation occurred de novo. An abnormal hGH mRNA lacking a region encoded by exon 3 was spliced when the mutant GH-I gene was expressed in cultured cells. Since skipping of exon 3 is a common feature for four different mutant GH-I genes identified in patients with autosomal dominantly inherited IGHD, we conclude that the mutation causes IGHD in this case.     

Influence of piecemeal necrosis on the compensatory increase of mitochondrial respiratory enzymes of the tumour-bearing liver: Clinical study of 53 surgical cases

The relationships between histological findings, adaptively increased cytochrome a(+a3) levels in chronic liver disease and complications after hepatectomy were studied in order to clarify the mechanism of mitochondrial derangement. The liver specimens of 53 hepatectomized patients were randomly evaluated by three independent hepatopathologists and were compared with cytochrome a(+a3) levels in the biopsied liver, the extent of operation and postoperative complications. The cytochrome a(+a3) concentrations did not show any significant difference between cases of chronic hepatitis and liver cirrhosis nor groups classified by regeneration. Severity of piecemeal necrosis was categorized into three groups: group A--minimal (n = 20); group B--moderate (n = 19); and group C--severe (n = 14). There were significant differences (P less than 0.01) in cytochrome a(+a3) concentrations between the groups (A: 99 +/- 9; B: 135 +/- 6; C: 155 +/- 10 pmol/mg of mitochondrial protein). Extensive hepatectomy, involving segmentectomy or more, was frequently complicated (four of nine, 44.4%) in group C, whereas there were few complications (two of 16, 12.5%) in group A cases in which extensive hepatectomy was performed. Evidence will be presented which will show that deranged liver function, as indicated by cytochrome a(+a3) levels, is closely correlated with piecemeal necrosis. This may be attributed to the damage of periportal hepatocytes which are the main sites of oxidative phosphorylation.     

Development of a rapid semi-quantitative immunochromatographic assay for serum hepatocyte growth factor and its usefulness in acute liver failure.

Measurement of serum human hepatocyte growth factor (HGF) by enzyme-linked immunosorbent assay (ELISA) is useful for the early diagnosis and prediction of prognosis of patients with acute liver failure (ALF). This ELISA methodology, however, is neither rapid nor convenient for use at the bedside. In this study, we have developed a rapid semi-quantitative immunochromatographic (IC) assay and evaluated its usefulness in assessing patients with acute hepatic injury. Only 100mul of serum is required; the assay can be easily completed in 20min. The values obtained using this novel assay correlated well with the values obtained using the standard ELISA protocol. In addition, the values obtained in the IC assay correlated with clinical course; increased serum HGF levels were associated with an increased frequency of ALF and death. These results indicate that this rapid semi-quantitative IC assay for HGF is useful for the early diagnosis of ALF and prediction of clinical outcome in acute hepatic injury.