Efficacy and side effects of intermittent recombinant interferon-alpha 2a in chronic aggressive hepatitis C: with or without initial daily administration.

To investigate the therapeutic effect and incidence of side effects of recombinant interferon-alpha 2a (IFN-alpha) in chronic aggressive hepatitis C under stratification by administration mode, a study was conducted by assigning patients to either group A (daily consecutive administration of 9 million units (MU) IFN-alpha for 2 weeks and, thereafter, 3 MU intermittently 3 times weekly for 22 weeks) or group B (exclusively intermittent administration; 9 MU IFN-alpha twice weekly or 6 MU IFN-alpha thrice weekly for 24 weeks). The 28 patients in group A received IFN-alpha for 24 weeks up to a total dose of 324 MU and the 53 patients in group B received the same for 24 weeks up to a total dose of 432 MU. When recovery was defined as the absence of hepatitis C virus (HCV)-RNA 6 months after the completion of treatment, the rate of recovery for group A was 32.1% and that for group B was 37.7%, the latter being higher but without significance. Side effects in groups A and B consisted of leucopenia occupying 14.3 and 7.5%, respectively, and thrombocytopenia occupying 42.9 and 11.3%, respectively; group B exhibited lower values for both side effects. No difference was detected between these groups in other side effects, including pyrexia, generalized malaise, arthralgia or psilosis. Intermittent administration from the outset permitted shortened duration of hospitalization and earlier rehabilitation. Intermittent administration of INF-alpha is required when treating patients with chronic hepatitis C showing lower leucocyte or platelet counts.

Serum antibody against Helicobacter pylori assayed by a new capture ELISA.

We developed a highly specific detection technique for serum antibody, using a monoclonal antibody to a specific antigen of Helicobacter pylori. A monoclonal antibody preparation that reacted with the 54-kDa molecule of H. pylori antigens was obtained. Using this preparation, an antigen-capture enzyme-linked immunosorbent assay (ELISA) was established by fixation of the monoclonal antibody, followed by reaction with sonicated whole cell antigens. The serum antibody titers of patients with gastritis and peptic ulcers were significantly higher than control titers, and the antibody titer correlated with the histological severity of gastritis. Patients positive for H. pylori by bacterial culture had higher titers than H. pylori-negative patients. Our new ELISA may be useful for the diagnosis of H. pylori infections and for evaluation of the severity of gastric inflammation.

Spontaneous complete regression of high grade non-Hodgkin's lymphoma. Morphologic, immunohistochemical, and gene amplification analyses.

BACKGROUND: Spontaneous regression of non-Hodgkin's lymphoma, occasionally reported in low grade groups, is a rare phenomenon in high grade groups. Clonal proliferation has not been confirmed in the majority of reported cases. In this woman, age 58 years, who had been diagnosed as having high grade immunoblastic lymphoma after excision of a single cervical lymph node, the remaining bilateral cervical, inguinal, and axillary adenopathy regressed completely without any cytotoxic treatments 22 days after biopsy. At the time of this writing, the patient has been free of disease for 24 months. METHODS: Clonality of the lymphoproliferation in the case was examined by immunohistochemistry and polymerase chain reaction (PCR) amplification using paraffin embedded biopsy material. Possible implications of Epstein-Barr virus in the pathogenesis of this process was examined also by PCR amplification and in situ hybridization. RESULTS: The proliferating lymphoid cells showed restricted expression of immunoglobulin (Ig) light chain and amplification of clonally rearranged V-D-J regions of Ig heavy chain gene. Epstein-Barr virus did not appear to be involved in the process. CONCLUSION: The present study shows that spontaneous complete regression of clonal lymphoproliferation that is morphologically a high grade lymphoma can occur.

[Antibacterial activity of cefdinir and omeprazole against Helicobacter pylori and their inhibition on H. pylori-producing urease].

Since Helicobacter pylori is isolated very frequently from gastric ulcer specimens, the combination therapy of antimicrobial agent and proton pump inhibitor has recently been used. A study was made on whether cefdinir (CFDN), amoxicillin (AMPC), metronidazole (MNZ), omeprazole (OPZ), and omeprazole-M (OPZ-M) have antimicrobial activity against H. pylori and whether they can inhibit H. pylori-producing urease. 1) CFDN, AMPC and MNZ showed a potent antimicrobial activity against H. pylori, and especially, AMPC showed a marked bactericidal activity in a short time. 2) OPZ is reported to be converted to OPZ-M, and active form, in the body. OPZ and OPZ-M showed a moderate antimicrobial activity against H. pylori, and scarcely any bactericidal activity. 3) CFDN and OPZ or AMPC and OPZ in combination did not show any synergistic effect on the antimicrobial activity, but MNZ and OPZ in combination showed additive effect on the antimicrobial activity against H. pylori. 4) OPZ and OPZ-M inhibited H. pylori-producing urease and the inhibitory effect of OPZ-M was more stronger than that of OPZ. CFDN, AMPC and MNZ did not show any inhibitory effect on H. pylori-producing urease at 10 micrograms/ml. From these data, antimicrobial agents and proton pump inhibitors in combination are expected to exert the in vivo synergistic effect since these drugs eradicate H. pylori and inhibit H. pylori-producing urease.

Concentration of serum laminin and type IV collagen in liver diseases assayed by a sandwich enzyme-immunoassay using monoclonal antibodies.

Serum laminin (P1 fragment) and type IV collagen levels were determined in patients with hepatic disorders. The method was based on a sandwich enzyme-immunoassay using two monoclonal antibodies that recognize different epitopes of either laminin or type IV collagen molecule. Laminin and type IV collagen levels in the serum of patients with chronic hepatic disorders were higher as compared with those in healthy control subjects, with the increment of serum type IV collagen being far greater than that of laminin. Since type IV collagen and laminin are major basement membrane components, it is suggested that the higher levels of these peptides may reflect a so-called capillarization of the perisinusoidal wall encountered in hepatic fibrogenesis. The assay system used in this experiment is simple and sensitive and can be applied to clinical evaluation of hepatic fibrosis.

Studies on macromolecular activators of phagocytosis from platelets (MAPPs) using anti MAPP murine monoclonal antibodies.

1-MAPP(290 kD) and s-MAPP(150 kD) are glycoproteins which have been shown to be involved in released products from platelets (PRPr) and to activate monocyte and neutrophil phagocytosis via the Fc receptors. Three IgG 2a (kappa) murine monoclonal antibodies against 1-MAPP and nine IgG1 (kappa) antibodies against s-MAPP have been raised. By affinity chromatography using these monoclonal antibodies, active MAPPs were obtained from the supernatant of outdated platelet concentrates in the presence of 60% saturated ammonium sulphate and from PRPr-rich medium, and substantial amounts of non-functioning immunoreactive MAPPs were obtained from the precipitate. Fab fragments of one of the two anti 1-MAPP and of two of the four anti s-MAPP monoclonal antibodies inhibited the function of their corresponding MAPPs, but none of them could inhibit the other MAPPs. 1-MAPP and s-MAPP in the platelets were visualized by indirect fluorescent antibody technique using Fab fragments of the antibodies.

Escape of phagocytosis-activating function from platelets into plasma during storage.

Stimulation of the phagocytic activity of leukocytes by two glycoproteins, the macromolecular activators of phagocytosis 1-MAPP and s-MAPP, involved in platelet release products (PRPr), was not observed when platelet concentrate (PC), stored longer than 48 h, was used as a platelet source. The supernatant of the PC (Sup) acquired this activity during storage. Salting out with ammonium sulfate (AS) revealed that the phagocytosis-enhancing activity was recovered in the supernatant of Sup from PC with 60% saturated AS, and the reverse activity, phagocytosis suppression, was observed in the precipitate with 30% AS. Suppressing activity was observed in PRPr prepared from both fresh and 48-h-old PC. The phagocytosis-enhancing activity observed in the Sup from the 48-hour PC was characterized by Sephadex G200 gel filtration and consisted of two peaks (300 and 150 kD), matching these of l-MAPP and s-MAPP in the PRPr prepared from fresh platelets. These results indicate that l-MAPP and s-MAPP escape from platelets into the plasma during storage, whereas the suppressing activity is retained in platelets for at least 48 h.

Purification and characterization of macromolecular phagocytosis activators released from platelets.

Substantial amounts of macromolecular activators of phagocytosis from platelets (MAPPs) were released in response to exposure of platelets to the specific agonists thrombin and collagen and to calcium ionophore A23187. Centrifugation of the platelets in culture medium also resulted in a release of MAPPs, but not when the platelets were frozen and thawed prior to centrifugation. In an experiment using outdated platelet concentrates, larger and smaller MAPPs (1-MAPP and s-MAPP, respectively) were purified from the thrombin stimulated supernatant (PRPr-plasma) by sequential steps of ammonium sulfate precipitation, delipidation with ethylacetate, ConA-Sepharose affinity chromatography, MONO Q anion exchange chromatography, and Superose 12 gel filtration. This procedure yielded 59,500-fold and 63,600-fold purified 1-MAPP (0.95 mg) and s-MAPP (0.41 mg), respectively, from 1,320 ml PRPr-plasma containing 84,500 mg protein. By gradient polyacrylamide gel electrophoresis the respective molecular weights (MW) of 1-MAPP and s-MAPP were 290-320 kd and 140-160 kd; isoelectric points were 5.6 and 4.6. Both MAPPs activated neutrophil phagocytosis of IgG-SRBC at concentrations in the range of 1 pM-100 nM. Indirect enzyme-linked immunosorbent assay (ELISA) and comparisons of the concentrations required for phagocytosis activation suggested that the MAPPs were not derived from fibronectin.

[Introduction of a method of promoting periodontal disease awareness].

In periodontal diseases, due to lack of subjective symptoms, disease awareness and professional consultations are often dismissed even by subjects with already established periodontal lesions. The purpose of this study was to gain data concerning the relationship between bleeding induced by interdental stimulating and some of the clinical indices used in routine periodontal examinations in order to consider if wooden interdental cleaners, which are available to patients themselves, could be used as a means of arousing disease awareness. Furthermore, the buccolingual insertion of interdental cleaners was confirmed together with reconsiderations on its role as a means of proximal plaque control. 180 interdental sites of 30 new out-patients suspected of gingivitis or periodontitis consisted the material. As comparative indices to interdental stimulating, Bleeding on Probing (BOP), Probing Depth (PD), Gingival Index (GI) and Gingival Crevicular Fluid (GCF) measurement were selected. The interdental cleaning efficiency was reconsidered by assessment of interproximal plaque removal. As a result, bleeding was induced both by interdental stimulating and probing in 106 of the 180 experimental sites and was the majority. In relation to PD, concerning pockets less than 1.5 mm, bleeding was not induced by interdental stimulating. 58.8% of 3 mm pockets and all of the pockets deeper than 5 mm bled on interdental stimulating. Concerning the relationship with the GI, no site evaluated GI. 0 bled on interdental stimulating. 12.9% of sites evaluates GI. 1, 73.3% of sites evaluated GI. 2, and all of the sites evaluated GI. 3 bled on interdental stimulating. 39.0% of sites with GCF measurement of 0-21, 51.9% of sites with GCF measurement of 21-40 and 96.8% of sites with GCF measurement of over 81 showed bleeding on interdental stimulating.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell surface heparan sulphate and adhesive property of sublines of rat ascites hepatoma AH7974.

Two variants (74AD and 74FL) established from rat ascites hepatoma AH7974 were examined for the production of glycosaminoglycans in culture. There was no difference between the adhesive (74AD) and the floating (74FL) variants in quantity of glycosaminoglycans produced by their cultivation in minimum essential medium supplemented with 10% foetal calf serum. However, they were distinctly different in the distribution patterns of heparan sulphate. In 74FL, about 70% of total heparan sulphate was found in the culture medium in soluble form, whereas in 74AD, only 7% was found in the medium and the rest was in the cell-substratum complex. In a serum-free medium, 74AD cells grew without adhering to the substratum. After cultivation, more than 90% of total heparan sulphate was found in the cell-associated fractions and the rest in the substratum fractions. No heparan sulphate was detected in the culture medium. On the other hand, 74FL cells released heparan sulphate to the serum-free medium as much as to the serum-containing medium. The increase in amount of heparan sulphate in the culture medium of 74FL cells was supposed to be caused by failure of the cells to deposit heparan sulphate at the cell surface and not caused by increased production. Cell-substratum adhesion mechanisms involving cell surface heparan sulphate (heparan sulphate proteoglycan) and some serum intermediate(s) are discussed for 74AD cells.

In vitro control of neutrophilic phagocytosis of IgG-coated SRBC by macromolecules involved in released products from platelets.

Analysis of macromolecular fraction of platelet release products (PRPr) was undertaken after we observed that pretreatment of human neutrophils with macromolecules (greater than 10(5) daltons) involved in PRPr enhanced phagocytosis of IgG-sensitized sheep red blood cells (IgG-SRBC). Sephadex G-200 gel filtration of PRPr revealed two macromolecular activators and two suppressors of neutrophilic phagocytosis of IgG-SRBC: MAPP(s), macromolecular activator(s) of phagocytosis from platelets; MSPP(s) macromolecular suppressor(s) of phagocytosis from platelets. The estimated molecular weights of the respective MAPPs were 100 kdaltons to 140 kdaltons (kd) (s-MAPP) and 200 kd to 290 kd (I-MAPP) and those of the respective MSPPs were 60 kd to 100 kd (s-MSPP) and 400 kd to 500 kd (I-MSPP). It is suggested that an I-MAPP molecule has at least an s-MAPP molecule as a functional unit and that MAPPs and larger MSPP (I-MSPP) might be glycoproteins, while smaller MSPP (s-MSPP) seems to contain neither carbohydrates nor lipids. As far as the effects of serum and plasma on phagocytosis of IgG-SRBC by neutrophils are concerned, the serum gave twice as much phagocytic activity as the plasma. Substantial MAPP-like activities were observed in the serum and MSPP-like activities were observed in both the serum and plasma.