Expression of ecalectin, a novel eosinophil chemoattractant, in nasal polyps.

CONCLUSION: Ecalectin, which is produced in the mucosa of nasal polyps, seems to play an important role in the accumulation and activation of eosinophils in nasal polyps, regardless of the presence or absence of atopic predisposition. OBJECTIVE: Ecalectin is a recently discovered eosinophil chemoattractant which elongs to the galectin family. We investigated the expression of ecalectin in nasal polyp tissues associated with various nasal and paranasal diseases in order to clarify the pathogenesis of eosinophilia in nasal polyposis. MATERIAL AND METHODS: Nasal polyps were taken from 56 patients diagnosed as having chronic sinusitis with nasal polyposis. The surgically resected polyps and nasal turbinates were immunohistochemically stained using antibodies against EG2, human mast cell tryptase, CD3 and ecalectin. RESULTS: The number of EG2- and ecalectin-positive cells was significantly higher in nasal polyps than control turbinates. Ecalectin-positive cells were observed in the subepithelial layer, where many EG2-positive cells were present. The number of ecalectin-positive cells correlated significantly with the number of EG2-positive cells in nasal polyps. Many ecalectin mRNA-positive cells were also observed in nasal polyps with an accumulation of EG2-positive cells.

Immunoanalytical detection of O4-ethylthymine in liver DNA of individuals with or without malignant tumors.

The detection and quantitation of carcinogen-DNA adducts in human cells are the key parameters in the molecular dosimetry of human exposure to environmental carcinogens. For investigating the possible relevance of alkylating N-nitroso compounds as causative agents in human carcinogenesis, we have quantitated O4-ethyl-2'-deoxythymidine (O4-EtdThd) in human liver DNA obtained from 33 autopsy specimens, i.e., 13 cases with primary liver cancer (LC), 8 with cancers other than liver cancer (OC), and 12 with noncancerous diseases (NC). None of the cases analyzed had a history of known occupational exposure to ethylating agents. The detection limit for O4-EtdThd was 3 X 10(-8) as a O4-EtdThd/dThd molar ratio in DNA, which was attained by the combination of prefractionation of DNA hydrolysates (= 20 mg of DNA/sample) by high performance liquid chromatography and competitive radioimmunoassay using anti-(O4-EtdThd) monoclonal antibody ER-01. Except for one case in each group, O4-EtdThd [or, alternatively, (an) unidentified structural modification(s) of DNA recognized by monoclonal antibody ER-01] was detected at mean (+/- SD) O4-EtdThd/dThd molar ratios of 39.9 +/- 40.2 x 10(-8), 53.5 +/- 74.0 X 10(-8), and 11.7 +/- 6.5 X 10(-8), respectively, in LC, OC, and NC. The difference of the O4-EtdThd content in DNA between LC and NC, or between LC + OC and NC, was statistically significant at P less than 0.05. These results suggest that humans are exposed to ethylating agents in vivo and that a premutagenic DNA lesion (O4-EtdThd) eventually accumulates in DNA, possibly to a biologically significant extent.

Heat-shock proteins and nucleolar hypertrophy in the liver of rat infused with methionine-free total parenteral nutrition.

Infusing a methionine-free solution into rats for 7 days resulted in a marked enlargement of liver nucleoli. By the analysis of two-dimensional gel electrophoresis, a spot 'a' (76 kDa, pI 5.3) stained with Coomassie blue was observed to accumulate highly in liver cytosol from rat infused with methionine-free solution. Metabolically labeling experiments with [35S]methionine showed that 'a' was more heavily labeled in primary hepatocytes of rats infused with methionine-free solution than in those of control rat. To ascertain whether 'a' is one of stress proteins, primary hepatocyte cultures were incubated at 42 degrees C for 2 h. 'a' (76 kDa, pI 5.3) was slightly induced in control hepatocytes but not appreciably in hepatocytes from the treated rat. In contrast, two other spots 'b' (74 kDa, pI 5.6) and 'c' (74 kDa, pI 5.3) were highly induced at 42 degrees C in hepatocytes from control and treated rats. The antibody against the consensus sequence peptide of hsp70 family reacted with 'a' (76 kDa, pI 5.3) as well as 'b' and 'c'. Immunoblot analysis revealed that 'a' accumulates highly in hepatocytes of treated rats. These results indicate that infusion of methionine-free solution into rats induces one member of the hsp70 family in hepatocytes.

Are there histopathologic characteristics particular to fulminant hepatic failure caused by human herpesvirus-6 infection? A case report and discussion.

An 8-month-old boy was admitted to a neighboring hospital for severe liver dysfunction and drowsiness 4 days after a diagnosis of exanthem subitum. A diagnosis of fulminant hepatic failure was made, and liver biopsy was performed during the acute stage. The presence of human herpesvirus-6 variant B (HHV-6B) DNA was shown in liver tissue by polymerase chain reaction (PCR) and in the endothelium of the portal vein by in situ hybridization (ISH). Histologic examination showed microvesicular steatosis resembling that of Reye's syndrome, even though aspirin had not been prescribed. We considered HHV-6 to be the causative agent in this case and report what is perhaps the first precise histologic description of fulminant hepatic failure caused by HHV-6.

HPV type 16 in conjunctival and junctional papilloma, dysplasia, and squamous cell carcinoma.

AIMS: To clarify the role of human papillomavirus (HPV) infection in the development of papilloma, dysplasia, squamous cell carcinoma, and basal cell epithelioma arising from the eyelids, including the tunica conjunctiva palpebrum (conjunctiva), its junction to epidemis of eyelid skin (junction), and eyelid skin. METHODS: Sixteen cases of papilloma, four of dysplasia, four of squamous cell carcinoma, and 12 of basal cell epithelioma were examined using formalin fixed and paraffin embedded samples. Detection of HPV-DNA was performed by PCR-RFLP and in situ hybridisation (ISH) methods. RESULTS: HPV-16 was detected in 12/16 papillomas (75%), 2/4 dysplasias (50%), and 1/4 squamous cell carcinomas (25%) but in none of the basal cell epitheliomas. No other HPV subtypes were found. ISH assay showed positive signals in only two cases of dysplasia and squamous cell carcinoma. The mean age of HPV-16 positive dysplasia and squamous cell carcinoma cases (81.7 years) was significantly higher than that of HPV-16 positive papilloma cases (p < 0.01). CONCLUSIONS: Based on the presence of HPV-16 in both benign and malignant lesions and the age distribution, it seems likely that HPV-16 alone may be incapable of causing development of conjunctival and junctional dysplasia and squamous cell carcinoma, and that any correlation between the papilloma-squamous cell carcinoma sequence and HPV infection may be due to rare events.

Systemic lupus erythematosus with multiple perivascular spongy changes in the cerebral deep structures, midbrain and cerebellar white matter: a case report.

A 42-year-old woman with systemic lupus erythematosus developed an episode of tonic seizures and progressive disturbance of consciousness at the terminal stage. Neuropathological examination of the brain revealed a nearly symmetrical distribution of multiple spongy foci in the internal capsules, thalami, globus pallidus, mesencephalic tegmentum, cerebral peduncles and hilus of the dentate nuclei. The spongy lesions were obviously distributed along apparently intact medium-sized veins, and contained large numbers of macrophages, and axonal spheroids and a few reactive astrocytes, without inflammatory cell infiltration. In addition, the perivenous spongy lesions exhibited IgG immunoreactivity, so it is surmised that some neurotoxic factor(s) that exuded from the veins in the center of the perivenous lesions may have brought about such a unique pathology.

Epstein-Barr virus infection resembling autoimmune hepatitis with lactate dehydrogenase and alkaline phosphatase anomaly.

A 73-year-old man had fever, lymphadenopathy, granulocytopenia, thrombocytopenia, ascites, pleural effusion, liver injury, and an allergic-like skin rash. Autoantibodies, such as anti-nuclear antibody, were shown, and there were lactate dehydrogenase and alkaline phosphatase anomalies and platelet-associated IgG. His liver injury resembled that in autoimmune hepatitis. He was diagnosed with Epstein-Barr virus (EBV) infection associated with autoimmunization because of his clinical course, fluctuation of anti EBV antibodies and positive EBV genome in circulating lymphocytes and serum. This case suggests a close relationship between EBV infection and autoimmunization or autoimmune-like hepatitis.

[Establishment and characterization of a tumor marker producing cell line (JR-1) derived from a gastric scirrhous cancer].

Despite the importance of in vitro study of gastric cancer, the established cell lines derived from human gastric carcinoma are very few. We have recently established a new cell line derived from human gastric cancer which has the ability to produce several tumor markers. This cell line has been designated JR-1. The cancer cells were obtained from the cerebrospinal fluid of a 37 year-old female patient who had metastatic brain tumor of the poorly differentiated gastric adenocarcinoma. The cells were inoculated into the tissue culture flask containing Ham's F-12 medium supplemented with 10% fetal bovine serum, antibiotics. Within 24 hours, the cells attached to the surface of the flask and started to grow. The first subcultures were performed at 1 week, and subsequent subcultures have been done once a week. This cell line has been maintained for more than 15 months through 60 passages with a stable growth. Chromosome analysis of the cells was performed. The doubling time of the 20th passage was 72 hours. Under phase contrast microscopy, monolayered pavement-like cell arrangement was observed. PAS staining showed intracellular mucin granules. Transmission and scanning electron microscopy revealed spindle-shaped cells with numerous microvilli and fine projections as well as intracellular granules, indicating mucin. Tumor markers produced by this cell line were CEA, CA19-9, TPA and Procollagen III.

Direct correlation between sonography and histology of minute malignant hepatic nodule.

To improve the qualitative diagnosis of ultrasonography (US) of minute malignant hepatic nodules, we studied the accurate and direct correlations between US and histopathology of 33 lesions using formalin-fixed autopsied livers. The target nodules were cut on the same plane as their US-image. The solid nests of malignant tumor without degeneration were hypoechoic on US. Hepatocellular carcinoma showed a wide variety of echo levels, from hypoechoic to isoechoic, and its images were more heterogeneous than those of other malignant lesions. The squamous cell carcinoma ranged from hypoechoic to hyperechoic according to the degree of keratinization. US may be useful for evaluating the degree of differentiation in squamous cell carcinoma. Marked mucin-production and "comedo" pattern seem to be histological factors of the hyperechoic US image of adenocarcinoma. This study may contribute to improve the quality of US-diagnosis of minute hepatic nodular lesions.

Proliferative response of human endothelial cultures to various types and treatments of human sera, to culture treatments and to various oxygen concentrations.

Endothelial cells from the human umbilical vein were exposed to different human sera, differently treated serum, to various oxygen concentrations, and various culture treatments. Endothelial proliferation was determined by measuring the uptake of [3H]-thymidine by means of autoradiography and presented as thymidine labeling index (TI) values. TI values differed according to different serum concentrations, serum types, serum preparations (WBS-PDS) and serum pretreatments. Low oxygen concentrations in the incubator atmosphere induced an early DNA synthesis response without an increase in cell number. The addition of the protease inhibitors aprotinin and soybean trypsin inhibitor to the culture medium resulted in a dose dependent TI decrease, whereas PMSF showed no influence.

The effect of CV-3988 and CV-6209 on the acute gastric erosions of rats due to water-immersion and restraint stress.

CV-3988 and CV-6209 inhibited gastric erosions in rats due to water-immersion and restraint stress in a dose-dependent manner. The above inhibitory effects of CV-3988 were observed in the presence of indomethacin, which may indicate that the inhibition is not prostaglandin dependent. The studies indicate that platelet-activating factor may be involved in the formation of erosions in rats under water-immersion and restraint stress.

Organ distribution of 3H-endotoxin in rats with liver fibrosis and rats with liver cirrhosis.

The organ distribution of 3H-endotoxin was investigated in rats with CCl4-induced liver injury. Wistar male rats were given water containing phenobarbital (Controls), or were treated with water containing phenobarbital and CCl4 inhalation. Rats inhaling CCl4 for 6 weeks developed liver fibrosis (Group LF), while those inhaling it for 10 weeks developed liver cirrhosis (Group LC). Animals were killed and examined 24 hours after an intravenous injection of 3H-endotoxin (12,000 CPM/l g body weight). Compared with the control rats, the measured amount of 3H-endotoxin per unit weight of spleen, lungs, and blood increased, while that of the liver significantly decreased in the rats of groups LF and LC. These results suggest that endotoxemia may be enhanced by a diminished uptake of endotoxin by the liver in liver fibrosis and liver cirrhosis.

Fate of 3H-labeled endotoxin in partially hepatectomised rats.

Organ distribution of endotoxin was compared in normal and previously hepatectomised rats after administration of endotoxin lipopolysaccharide labeled at the galactose moiety. Most of the radioactivity was found in the liver in both cases, but total recovery was much higher in the organs of normal rats. In the liver, the amount of endotoxin per total organ and per 1 g organ weight was significantly reduced in the hepatectomised rats as compared with the intact controls. In normal rats, the radioactivity decreased 12 to 24 hours in the liver, but in the hepatectomised rats, no significant change was observed during this period. The total radioactivity in the lungs, spleen and blood (1 ml), as well as that per unit weight bases, was significantly higher in the hepatectomised rat than in normal rats. These results and autoradiographical studies suggest that endotoxin clearance activity of Kupffer cells in the liver is decreased by partial hepatectomy of the rats at the peak phase of regeneration, and that remaining endotoxin flow out of the liver is disposed mainly by the macrophages in the spleen.

[Dystrophin abnormality in a patient with polymyositis associated with primary biliary cirrhosis and myocardial injury].

We have reported a 58-year-old Japanese female with polymyositis, primary biliary cirrhosis (PBC) and arrhythmia. In contrast to the previously reported 13 cases of polymyositis associated with PBC, symptoms and laboratory data abnormalities responded to oral administration of predonisolone. Interestingly, immunohistochemical and immunoblot analyses of biopsied skeletal muscle revealed diminished expression of dystrophin carboxyl-terminal domain in the sarcolemma, suggesting that, in analogy to Duchenne muscular dystrophy, secondary abnormality of the link between the basal lamina and cytoskeleton via the dystrophinglycoprotein complex may have played a role in the molecular pathogenesis of muscle fiber degeneration in this patient.

Anti IgE-induced asthma in guinea pigs; a new model for asthma. Physiological and pathological studies.

We established an experimental animal model for IgE-mediated asthma by the use of intravenous injection of rabbit anti guinea pig IgE in normal guinea pigs. The degree of asthmatic airway obstruction was quantitatively estimated by Mead's apparatus which enabled us to use unanesthetized guinea pigs. With the injection of anti IgE, only an early asthmatic response was observed. Inhalation of anti IgE did not trigger asthma. Histo-pathological changes in the lung obtained immediately after the injection of anti IgE resembled those reported in human asthma. This system may be used for pathological and pharmacological studies of the IgE-dependent early asthmatic response.

[Histopathological analysis of 73 autopsy cases of "acute-on-chronic" in Japan--influence of alcohol abuse on exacerbation of chronic liver disease].

Cases of acute exacerbation in chronic liver disease ("acute-on-chronic") in Japan were surveyed by questionnaire method since 1986 and 220 cases were collected. 73 pathological specimens of 102 cases which were autopsied were subjected to morphological analysis. The cases were divided according to causes of acute exacerbations: namely super-imposed viral infection or drugs (Group I), unknown causes or post immunosuppressant therapy (Group II), alcohol abuse (Group III), operation, digestive tract hemorrhage or post TAE etc, (Group IV). Area of hepatic necrosis was large and regeneration of hepatocytes were significantly strong in Group I and II compared with cases of Group IV. Regeneration was also inhibited in liver of habitual alcohol drinker. Significant difference was shown in cases of Group IV. Pathologically liver of 10 cases of Group III divided to florried cirrhosis (3 cases), alcoholic liver cirrhosis (4 cases) and alcoholic hepatitis combined with post hepatitic liver cirrhosis (3 cases, all HBV carriers).

[Histopathological studies on paranasal mucosa from patients treated with erythromycin].

Low-dose and long-term administration of erythromycin (EM therapy) has been reported to be very effective for patients with intractable chronic sinusitis including sinobronchial syndrome. However, we sometimes encounter patients whose sinusitis is extremely resistant to EM therapy. Therefore, the present study was carried out to determine the correlation between the clinical efficacy of EM therapy and the histopathological features of the ethmoidal mucosa and nasal polyps of patients treated with erythromycin. Patients with significant lymphocytic infiltration in the submucosal area responded well to EM therapy. Furthermore, patients with neutrophilic infiltration within and beneath the ciliary epithelium tended to show improvement with this therapy. On the other hand, EM therapy was minimally effective in patients whose subepithelial layer showed marked eosinophilic infiltration. In patients showing excellent and good clinical results, histological changes in the paranasal mucosa after therapy were as follows: a decrease in the number of inflammatory cells, reduced interstitial edema, increased fibrosis and normalization of the secretory glands. From these results, we conclude that erythromycin may suppress chronic inflammation except for an allergic reaction characterized by marked eosinophilic infiltration.