Ultrasound evaluation of the fibrosis stage in chronic liver disease by the simultaneous use of low and high frequency probes.

A liver biopsy is currently considered the definitive diagnostic modality for establishing the severity of hepatic fibrosis. We analysed the diagnostic sensitivity and accuracy of ultrasound (US) using both low frequency and high frequency probes as a repeatable, inexpensive, and reliable method to determine the fibrosis stage in chronic liver disease and then compared our results with the histological findings. A total of 103 patients with chronic liver disease (60 males and 43 females, average age 51 years old) who had undergone both a liver biopsy and US with 2-5 MHz frequency and 5-12 MHz frequency probes were prospectively evaluated in this study. An US scoring system using both the low frequency and high frequency probes was performed by evaluating the edge, surface and parenchymal texture of the liver. Each score was obtained by evaluating three parameters; the bluntness of the liver edge, the irregularity of the surface and the coarseness of the parenchymal texture were evaluated and then compared with the histological findings. The US scores of the liver edge (rs: 0.6668), liver surface (rs: 0.9007) and liver parenchymal texture (rs: 0.8853) correlated significantly with the fibrosis stage obtained based on the biopsy findings. The accumulated US scores of these three parameters, however, was found to be the most reliable indicator (rs: 0.9524). Patients with an accumulated score of 6.5 or more were all found to have fibrosis stage 4 in which the accuracy of our scoring system for correctly predicting cirrhosis was found to be 100% sensitive. When an accumulated US score of 3 was interpreted to indicate mild fibrosis (a fibrosis score of 0 or 1), all 42 patients with stage 0 or 1 fibrosis were found to have an accumulated US score of 3 or less (a probability of 100%) and 42 of 53 patients with a score of 3 or less were found to have stage 0 or 1 fibrosis (specificity of 79.2%). An ultrasound evaluation of the liver fibrosis stage based on the scoring system using both low and high frequency probes was found to be a reliable and effective alternative to the histological staging in chronic liver diseases.

Skull metastasis of hepatocellular carcinoma associated with acute epidural hematoma: a case report.

BACKGROUND: Although epidural hematoma is well documented in trauma patients, its association with other etiologies, such as neoplasms, is not widely known. Here the authors report a case of acute epidural hematoma that originated from a metastatic hepatocellular carcinoma (HCC) in the skull. CASE DESCRIPTION: A 70-year-old male was admitted to our hospital with left-sided hemiparesis. Preoperative computed tomography (CT) revealed a lenticular high-density area adjacent to the right parietal bone, consistent with an acute epidural hematoma. A subsequent magnetic resonance image (MRI) showed a skull tumor adjacent to the epidural hematoma. Removal of the tumor and evacuation of the hematoma were performed and the pathological diagnosis was metastatic HCC. Postoperatively, the patient gradually recovered but he died of pneumonia 2 months later. CONCLUSION: This report represents an additional example of a rare case of metastatic skull tumor associated with acute epidural hematoma. The authors suggest that metastatic skull tumors may be one of the important differential diagnoses in patients with acute epidural hematoma.

Pharmacological modulation of LPS-induced MIP-1 alpha production by peripheral blood mononuclear cells.

In the present study, we investigated the effects of some anti-asthmatic drugs on the production of the CC chemokine, macrophage inflammatory protein-1 alpha (MIP-1 alpha), in response to lipopolysaccharide (LPS) by peripheral blood mononuclear cells (PBMC). MIP-1 alpha production was induced by LPS in a concentration-dependent fashion and reached the maximum at 10 micrograms/ml LPS (27.5 +/- 2.3 ng MIP-1 alpha/10(6) PBMC). At a submaximal concentration of LPS (1 microgram/ml), the release of MIP-1 alpha increased with time and reached the maximum 24 h after LPS stimulation. Actinomycin D and cycloheximide inhibited MIP-1 alpha production completely, but glucocorticoids did not completely inhibit MIP-1 alpha production, with a maximum inhibition of 70%. We examined the effect of beta-stimulants and phosphodiesterase inhibitors, which upregulate intracellular cyclic AMP levels, on MIP-1 alpha production. When PBMC were treated with beta-stimulants alone, beta-stimulants showed a slightly inhibitory effect on MIP-1 alpha production. However, the coadministration of roliplam significantly potentiated the inhibitory effect of beta-stimulants on MIP-1 alpha production. Moreover, db-cAMP suppressed MIP-1 alpha production dose-dependently. The above data indicate that the production of MIP-1 alpha is regulated by cyclic AMP and that cyclic AMP could provide a useful target for therapeutic treatment in asthmatic diseases and other diseases where MIP-1 alpha is involved in their etiology.

Relationship between sustained elevation of serum alanine aminotransferase and progression from cirrhosis to hepatocellular carcinoma: comparison in patients with hepatitis B virus- and hepatitis C virus-associated cirrhosis.

Most patients with hepatocellular carcinoma (HCC) in Japan have hepatitis B virus (HBV)-or hepatitis C virus (HCV)-associated cirrhosis. In the present study, the risk of HCC in patients with cirrhosis was analysed by the levels of serum alanine aminotransferase (ALT). One hundred and one (78%) of 129 patients with cirrhosis registered from April 1979 were followed at monthly intervals with the measurement of serum ALT. Of 101 patients, 38 tested positive for hepatitis B surface antigen (HBsAg) but negative for antibody to HCV (anti-HCV; HBV group), 47 tested negative for HBsAg but positive for anti-HCV (HCV group) and nine tested positive and seven tested negative for both. Mean serum ALT during follow-up was calculated on the basis of monthly values during the observation period that started at enrolment and ended with the detection of HCC or at the end of March 1994. By the end of March 1994, 37 (37%) patients developed HCC; 12 were in the HBV group, 21 in the HCV group and four were in the group positive for both. Mean serum ALT during the observation period was similar in patients who developed HCC and those who did not develop HCC in the HBV group. In contrast, the value was significantly higher in patients who developed HCC than in patients who did not develop HCC in the HCV group (P < 0.05).

Prevalence of hepatitis B or C virus infection in patients with fulminant viral hepatitis. An analysis using polymerase chain reaction.

BACKGROUND/METHODS: The cause of fulminant hepatitis is still not fully understood. We studied 23 patients with fulminant hepatitis, using polymerase chain reaction to detect hepatitis virus genomes. Tests for HBsAg and IgM anti-HAV and IgM anti-HBc were performed in all patients. Serum samples were stored at -70 degrees C for later analysis of anti-HCV and hepatitis virus genomes such as hepatitis B virus, hepatitis C virus and hepatitis D virus. RESULTS: Of 23 patients, 17 (74%) had HBV-DNA and two (9%) had HCV-RNA. No patient was positive for both viruses or positive for HDV-RNA. Serological tests indicated that two patients, negative for HBV-DNA and HCV-RNA, were positive for IgM anti-HAV. In contrast, 8 of 17 (47%) HBV-DNA positive patients were negative for both IgM anti-HBc and HBsAg in conjunction with low levels of viremia. Four patients were positive for anti-HCV, but only one was positive for HCV-RNA. One patient, positive for HCV-RNA, was negative for anti-HCV. CONCLUSIONS: Our results indicate that analysis of hepatitis virus genomes using polymerase chain reaction allows accurate identification of the virus causing fulminant hepatitis.

Detection of hepatitis B virus genome in hepatocellular carcinoma from both hepatitis B surface antigen- and antibody to hepatitis C virus-negative patients.

Twenty-two patients who tested seronegative for both hepatitis B virus (HBV) and hepatitis C virus (HCV) markers and undefined pathogenesis of hepatocellular carcinoma (HCC) were studied using polymerase chain reaction (PCR). Among these patients, 5 (23%) were positive for HBV-DNA in serum by PCR, but no patient was positive for HCV-RNA by reverse transcription PCR. Liver tissue specimens were available for analyzing HBV genome by PCR in 17 patients, and HBV sequences were detected in 10 (59%) patients. These results indicate that HBV is commonly implicated in serologically undefined pathogenesis of HCC in Japan.

Analysis of wild-type and e antigen-defective hepatitis B viruses during the course of a short-term corticosteroid therapy in chronic hepatitis B.

Hepatitis B virus (HBV), with a G-to-a point mutation at nucleotide 83 in the precore region (mutant HBV83), accounts for most cases of hepatitis B e antigen (HBeAg)-defective HBV. However, it is still not clear how mutant HBV83 is associated with HBe seroconversion. Twenty-six HBeAg-positive patients with chronic hepatitis B who received oral prednisolone (30 mg/day) for 3 weeks were studied to clarify the prevalence of mutant HBV83 during the treatment using polymerase chain reaction with a restriction fragment length polymorphism assay. Twelve (46%) patients seroconverted to anti-HBe 1 year after treatment, whereas 14 (54%) did not. The proportion of mutant HBV83 to whole HBV remained unchanged in both groups during an acute exacerbation induced by withdrawal of corticosteroids. Among 12 anti-HBe-seroconverted patients, five (56%) of nine patients with only wild-type HBV at baseline developed detectable levels of mutant HBV83 while all three patients with a mixed viral population of wild-type HBV and mutant HBV83 at baseline developed a higher proportion of mutant HBV83 one year after treatment. In contrast, these changes were observed in only one (14%) of seven who failed to seroconvert. The results indicate that a flare-up of hepatitis precedes emergence or selection of mutant HBV83, followed by HBe seroconversion in patients with chronic hepatitis B.

Changes in the prevalence of HBeAg-negative mutant hepatitis B virus during the course of chronic hepatitis B.

Hepatitis B virus with a G-to-A point mutation at nucleotide 83 in the precore region (mutant hepatitis B virus 83), which cannot produce HBeAg, is commonly found in HBe antibody-positive hepatitis B virus carriers. We analyzed the consecutive changes in the prevalence of mutant hepatitis B virus 83 during the course of chronic hepatitis B virus infection. Forty-five patients with chronic hepatitis B who were followed up for more than 2 yr in our hospital were studied by polymerase chain reaction in combination with a restriction fragment length polymorphism assay. Mutant hepatitis B virus 83 was found in 14 of 18 (78%) HBe antibody-positive patients and in 8 of 27 (30%) HBeAg-positive patients at baseline. Eighteen of the 22 patients who had mutant hepatitis B virus 83 (82%) showed mixed viral populations of wild-type hepatitis B virus and mutant hepatitis B virus 83, whereas 4 (18%) had only mutant hepatitis B virus 83 and were positive for HBe antibody. During a 2 yr follow-up period, mutant hepatitis B virus 83 was newly detected in 9 of 23 (39%) patients who had wild-type hepatitis B virus alone at baseline. The proportion of mutant hepatitis B virus 83 to whole hepatitis B virus in the serum of 18 patients with mixed viral populations at baseline fluctuated during follow-up. In contrast, wild-type hepatitis B virus was never detected throughout the study in all four patients who had only mutant hepatitis B virus 83 at baseline.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of NZ-107, a newly synthesized pyridazinone derivative, on antigen-induced contraction of human bronchial strips and histamine release from human lung fragments or leukocytes.

The effects of a newly synthesized pyridazinone derivative, NZ-107, 4-bromo-5-(3-ethoxy-4-methoxybenzylamino)-3(2H)-pyridazinone, and two well-known antiasthmatic drugs, amlexanox (orally active disodium cromoglycate-like drug) and disodium cromoglycate (DSCG) on antigen-, histamine- and leukotriene C4 (LTC4)-induced constriction of isolated human tracheal muscle, and histamine release from human lung tissues and leukocytes were investigated in vitro. In some experiments, salbutamol was used as a reference drug. NZ-107 inhibited antigen-, histamine- and LTC4-induced contraction of tracheal muscle. Amlexanox and DSCG did not affect the contractile response of tracheal muscle caused by each stimulant. Salbutamol inhibited antigen-induced contraction of tracheal muscle. NZ-107, amlexanox, DSCG and salbutamol clearly inhibited the antigen-induced release of histamine and LTC4 from human lung tissue. The antigen-induced histamine release from atopic human leukocytes was inhibited by NZ-107 and amlexanox, but not by DSCG. Pretreatment with IL-3 did not alter antigen-induced contraction of tracheal muscle and histamine release from lung tissue, but antigen- or calcium ionophore A 23187-induced histamine release from leukocytes was clearly enhanced. Amlexanox inhibited the IL-3-induced enhancement of histamine release from leukocytes in the case of both stimuli, but NZ-107 and DSCG had no effect. These data suggest that NZ-107 has potent anti-allergic actions based on the inhibition of antigen-induced contraction of human tracheal muscle and mediator release from human lung tissue and leukocytes.

Responses of isolated Japanese monkey tracheal muscle to allergic mediators.

The responsiveness of isolated Japanese monkey (Macaca fuscata) tracheal muscle to antigen, carbachol, histamine, leukotriene C4 (LTC4), U-46619 and substance P (SP) was compared to that of isolated human trachea. Weak but persistent contraction was observed after the addition of antigen to isolated Japanese monkey tracheal muscle passively sensitized with monkey serum containing IgE antibody against Japanese cedar (Cryptomeria japonica) antigen. Unlike monkey tracheal muscle, a fair amount of contraction was caused by the antigen in human tracheal muscle passively sensitized with human atopic serum. When chopped, passively sensitized monkey or human lung tissue was challenged with antigen, a significant level of histamine was released from these tissues. In Japanese monkey tracheal muscle, histamine and SP produced no contraction of tracheal muscle, whereas carbachol, LTC4 and U-46619 caused contraction in a dose-dependent fashion. Contrary to the Japanese monkey, histamine and carbachol caused distinct contraction in tracheal muscle obtained from the cotton-headed tamarin (Saguinus oedipus). In human tracheal muscle, all test substances (carbachol, histamine, LTC4, U-46619 and SP) induced clear contraction. In lung parenchyma obtained from Japanese monkey, histamine induced a weak contraction, and this histamine-induced contraction was also inhibited by pyrilamine (H1 receptor antagonist). These results indicate that antigen-induced contraction of isolated Japanese monkey tracheal muscle, passively sensitized with monkey atopic serum, is not a useful model for human allergic bronchoconstriction in vitro because of the unresponsiveness of tracheal muscle to histamine and SP.

The effect of ONO-3708, a novel TxA2 receptor antagonist, on U-46619-induced contraction of guinea pig and human tracheal strips in vitro and on bronchoconstriction in guinea pigs in vivo.

The effect of (9, 11), (11, 12)-didedoxa-9 alpha, 11-alpha-dimethylmethano-11,12-methano-13,14-dihydro-13-aza-14-oxo -15-cyclo-pentyl-16, 17, 18, 19, 20-pentanor-15-epi-TxA2 (ONO-3708) on 9,11-methanoepoxy-prostaglandin H2 (U-46619)-induced contraction of airway smooth muscle in the guinea pigs and human in vitro and bronchoconstriction in guinea pigs in vivo was investigated. In in vitro experiments, ONO-3708 inhibited the U-46619-induced contraction of isolated guinea pig and human tracheal smooth muscle in a dose related fashion (guinea pig; pA2=7.78, human; pA2 = 7.43). The contractions of guinea pig tracheal muscle caused by histamine and leukotriene D4 (LTD4) were not inhibited by ONO-3708. In in vivo experiments, intravenous injection of ONO-3708 at doses between 1 and 20 mg/kg inhibited the U-46619-induced increase of airway insufflation pressure as measured by Konzett-Rössler method. In addition, ONO-3708 inhibited the U-46619-induced increase in airway reactivity to acetylcholine. These data suggest that ONO-3708 has possible therapeutic utility for asthma in which TxA2 participates.

Inhibitory effects of beta-adrenergic stimulants on increased vascular permeability caused by passive cutaneous anaphylaxis, allergic mediators, and mediator releasers in rats.

The effects of isoproterenol, salbutamol, theophylline, and forskolin on IgE antibody-mediated homologous passive cutaneous anaphylaxis (PCA) and on skin reactions caused by allergic mediators and mediator releasers were investigated in rats. Isoproterenol and salbutamol dose-dependently inhibited the PCA and skin reactions caused by histamine, serotonin, and leukotriene C4 (LTC4), elicited at the same time in the same rat. These agents also dose-dependently inhibited the skin reactions caused by platelet-activating factor (PAF), leukotriene D4 (LTD4), compound 48/80 (48/80), and calcium ionophore A23187 (A23187). Propranolol overcame the inhibitory effect of isoproterenol on PCA and skin reactions in a dose-dependent manner. Theophylline and forskolin showed similar effects as the beta-adrenergic stimulants. These results indicate that beta-adrenergic stimulants inhibit the increased vascular permeability caused by allergic mediators, and suggest that this activity of beta-adrenergic stimulants might play an important role in their antiallergic actions. Inhibition of increased vascular permeability might be mediated via beta-receptors and may be related to the increase in intracellular cyclic AMP levels.

Solid phase enzyme-linked immunosorbent assay (ELISA) for anti-sheep erythrocyte antibody in mouse serum.

The solid phase enzyme-linked immunosorbent assay (ELISA) was developed to measure IgM and IgG anti-SRBC antibody titers in mouse serum. Sheep erythrocytes, which have a surface negative charge, were attached directly to the bottom of an aminoplate well (96-well flat bottom, Sumitomo Bakelite Co.) which is charged positively to provide a solid phase for the ELISA antigen. Serum samples titrated were obtained from mice 5 and 10 days after SRBC immunization. Alkaline phosphatase-labeled goat anti-mouse IgM and/or IgG preparations were used as second antibody. Alkaline phosphatase activity in a well was measured by the Kind and King method [Kind, P. R. N. & King, E. J. (1954). J. clin. Path., 7, 322-326]; the optical density (OD) value of quinone as a final product was measured at 492 nm. (1) A linear relationship was observed between the antiserum concentration and OD value over a wide enough dilution range to assay antibody titers of serum samples; (2) IgM and IgG fractions from antiserum showed almost the same antibody titer as did the reconstituted samples; (3) a good correlation was observed between the serum IgM titer and the number of IgM hemolytic plaque-forming cells (PFCs) in spleen 5 days after the immunization, and was also observed between serum IgG antibody titer and the number of IgG PFCs 5 and 10 days after. Therefore, the ELISA described here requires no fixative for preparation of cell-coated plates, and serum IgM and IgG anti-SRBC antibody titers can be measured without any fractionation technique.

Effects of different drugs on passive cutaneous anaphylaxis elicited in the mouse ear at 1.5 hours.

IgG1, antibody-mediated homologous passive cutaneous anaphylaxis at 1.5 h (1.5-hour PCA) was elicited in the ears of mice and the effects of different drugs were studied. The reaction, as measured by the amount of extravasated dye, was inhibited by antihistamines, antiserotonins, cyclic AMP-elevating agents, tranilast and ketotifen, but not by an SRS-A antagonist (FLP 55712), lipoxygenase inhibitors, cyclooxygenase inhibitors and disodium cromoglycate. These results suggest that the pharmacological profile of 1.5-hour PCA resembles that of 48-hour PCA mediated by IgE antibody in the mouse ear.

Effect of cinnarizine on IgE antibody-mediated experimental allergic reactions in guinea pigs.

The anti-allergic activity and mechanism of cinnarizine was investigated in guinea pigs. Nifedipine, a calcium antagonist, and tranilast, a potent, orally active anti-allergic agent, were used as comparative drugs. Cinnarizine protected against fatal systemic anaphylactic shock in guinea pigs passively sensitized with IgE antibody. Cinnarizine reduced many of the features of severe respiratory disorders. Nifedipine and tranilast showed similar effects. Cinnarizine and nifedipine inhibited the contractile response to antigen of sensitized tracheal smooth muscle when the challenge was carried out at low antigen concentrations. Tranilast showed a tendency to inhibit the antigen-induced contraction of tracheal smooth muscle. Cinnarizine and nifedipine inhibited Ca-induced contraction in potassium-depolarized tracheal smooth muscle, tranilast had no effect. Cinnarizine showed antagonistic action to the contraction by histamine or leukotriene D4 (LTD4) of tracheal muscle. Nifedipine showed similar antagonistic action, although its potency is lower than cinnarizine. Tranilast showed slight antagonistic action to LTD4. Antigen-induced release of histamine and slow reacting substance of anaphylaxis (SRS-A) from sensitized lung tissues was inhibited by nifedipine and tranilast but not by cinnarizine. The release of histamine and SRS-A from lung tissues by calcium ionophore A23187 was inhibited by nifedipine and tranilast but not by cinnarizine. These results suggest that the anti-allergic action of cinnarizine is mainly due to the antagonistic action to allergic mediators and not by interfering with the release of mediators. Cinnarizine's mechanism seems to be related to its antagonistic action to Ca in smooth muscle, but not to the transport of Ca in releasing the anaphylactic chemical mediators in mast cells and other target cells.

Specific suppression of antigen-antibody reactions by a dialysate from Dermatophagoides farinae.

Mite dialysate prepared from an extract of house dust mite, Dermatophagoides farinae, suppressed 48-h homologous passive cutaneous anaphylaxis (PCA) in rats caused by a non-dialyzable fraction of the mite extract (mite antigen), and the suppression was dose-dependent with a high specificity for the antigen. The mite dialysate itself slightly elicited the PCA. Fraction 3 obtained from the mite dialysate by means of DEAE-Sephadex A-25 column chromatography suppressed PCA in rats and guinea pigs more potently than mite dialysate. Histamine release from sensitized rat peritoneal exudate cells (PEC) induced by the mite antigen was suppressed in a dose-dependent manner when the PEC had been previously incubated with fraction 3. However, incubation of the PEC with fraction 3 caused a significant release of histamine compared to the spontaneous value. Fraction 3 clearly inhibited the passive hemagglutination of the mite antigen-conjugated sheep red blood cells caused by rat hyperimmunized serum. From these results it was considered that certain haptenic substances might be present in the mite dialysate.

Immunoglobulin E antibody production against house dust mite, Dermatophagoides farinae, in mice.

Immunoglobulin E (IgE) antibody production against Dermatophagoides farinae extract (mite antigen) was studied in female BALB/c mice. When mice were immunized with mite antigen and aluminium hydroxide gel (alum) intraperitoneally at intervals of 30 d, good primary, secondary and tertiary IgE antibody responses were observed. In the absence of adjuvant, a subcutaneous injection of mite antigen failed to induce the primary IgE antibody response. However, good IgE antibody responses were observed after the secondary immunization given 30 d after the primary immunization. Furthermore, 5 weekly injections of mite antigen alone also induced the IgE antibody production. Intranasal administrations of mite antigen alone also induced the IgE antibody production in mice. Two exposures to mite antigen, intranasally, were sufficient for eliciting low IgE antibody production. The response was significantly potentiated by the administration of islet-activating protein obtained from culture fluids of Bordetella pertussis. These results indicate that the intranasal administration of mite antigen is very effective for eliciting the IgE antibody production.