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Male and female adult C3H- +/+, C3H-gld/gld.lpr/lpr (gld.lpr) and CBA-lprcg/lprcg (lprcg) mice were given a single i.p. dose of 30 mg/kg dimethylnitrosamine (DMN). Liver tissues were collected from mice killed 6, 12, 24 and 36 hrs post treatment, and the progression of the lesions was characterized morphologically and by the TUNEL method. DMN induced centrilobular hepatic injury accompanied with acute hemorrhage, and all mice died 36 to 48 hrs after the dosing. At 12 hrs after DMN administration, centrilobular hepatocytes revealed nuclear chromatin clumping. At 24 hrs, hepatocyte nuclei became fragmented to form apoptotic cells. Ultrastructurally, chromatin was condensed into a compact granular mass or crescent granular cap at the nuclear periphery. At 36 hrs, the number of apoptotic cells increased and they protruded into the sinusoid or were engulfed by the neighboring hepatocytes. A TUNEL-positive signal preceded the morphological changes and a few normal appearing centrilobular hepatocytes were positive 6 hrs post dosing. Endothelial damage was seen immunohistochemically at 24 hrs by disruption of type IV collagen and factor VIII-related antigen, resulting in massive hemorrhage in the centrilobular to mid zone. No inflammatory reactions were observed throughout the degeneration. The findings indicate that a single i.p. administration of DMN induced severe and fatal toxicity in liver tissues in mice which resembled human fulminant hepatitis. However, as gld-lpr and lprcg mice defective in apoptosis through the Fas system also showed similar severe liver damage, the Fas/Fas ligand system is not involved in DMN-induced liver apoptosis. No other organs or tissues were damaged, and the control mouse liver was intact.

The histological changes occurring in the esophageal mucosa of shrews (Suncus murinus) after N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) treatment were investigated sequentially. Six-week-old female shrews were given a 50 micrograms/ml MNNG solution as drinking water for 30 weeks, and 5 selected at random were killed at 10 and 20 weeks of age, and thereafter at 5-week intervals until 45 weeks of age. Controls were killed at 45 weeks of age. The MNNG-induced esophageal lesion in shrews began from basal cell hyperplasia at 20 weeks of age, followed by dysplasia occurring at 25 weeks of age, then progressed toward intraepithelial carcinoma to invasive squamous cell carcinoma at 35 weeks of age. Apparent sequential dysplasia-carcinoma transition was seen. Papillomas were seen from 25 weeks of age but there was no evidence of papilloma-carcinoma sequence. Five MNNG-untreated shrews killed at the end of the experiment were free of esophageal tumors.

Female 6-week-old shrews were given a solution of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) at a concentration of 50 micrograms/ml or 100 micrograms/ml in the drinking water. All 11 shrews receiving 100 micrograms/ml MNNG died 8-13 days after the beginning of carcinogen administration and 6 of the 20 shrews receiving 50 micrograms/ml MNNG died after 10-54 days. When animals were between 43 and 54 weeks of age, multiple esophageal lesions were evoked in all 14 that had received 50 micrograms/ml MNNG for 30 weeks. All shrews developed a protruding, ulcerative, or superficial type of squamous-cell carcinoma of the esophagus, accompanied by papillomas. Local invasion was seen in squamous-cell carcinoma but no distant metastasis was noted. None of the 5 control shrews developed any esophageal abnormality. No gastric adenocarcinoma, intestinal sarcoma, or other tumors were induced with MNNG. It can be concluded that MNNG has a carcinogenic effect on shrew esophageal epithelium.

The carcinogenic effects of 1, 2-dimethylhydrazine (DMH) were investigated in virgin female Japanese house musk shrews, Suncus murinus (family: Soracidae, order: Insectivora). From 6 weeks of age, 15 shrews were given 14 doses of DMH (40mg per kg, s.c.), administered weekly (group 1) and another 15 animals were given 28 doses of DMH (20mg per kg, s.c.), also administered weekly (group 2); 5 untreated shrews served as controls. A high incidence of musk gland tumors, and some other tumors (carcinomas of the small intestine, pulmonary adenoma, liver adenoma, and leukemia) were induced in DMH-treated shrews, whereas no such tumors were seen in untreated shrews up to 60 weeks of age. Musk gland tumors developed in 82% (9/11) of animals in group I and in 92% (12/13) of animals in group 2 at 36 to 60 weeks of age. DMH, a colonotrophic carcinogen in rodents, did not evoke colon cancers in shrews.

The influence of various fixatives and fixation time on Proliferating Cell Nuclear Antigen (PCNA) immunoreactivity in paraffin-embedded sections using the rat ileum were compared, and the effect of antigen retrieval based upon microwave heating in the presence of a solution of saturated lead thiocyanate was investigated. PCNA immunoreactivity stained more intensely when tissues were fixed in Bouin's fluid for 6 hr or methacarn for 3 hr than in 10% buffered formalin or 4% para-formaldehyde for 3 hr to 1 week, respectively. In general, the number of PCNA positive cells and staining intensity decreased as the fixation time was prolonged. In sections fixed in formalin for over 24 hr, PCNA immunoreactivity was greatly reduced. Antigen retrieval increased the intensity of immunostaining in Bouin's, formalin and PFA-fixed tissues, whereas, methacarn-fixed tissues showed no enhancement. These results suggest that this technique may alter protein cross-linking caused by aldehydes.

Antigen retrieval based on microwave-exposure of formalin-fixed, paraffin-embedded tissues in a metal solution of saturated lead thiocyanate or 20% zinc sulfate was investigated. Six leiomyosarcomas and 4 rhabdomyosarcomas were studied using the ABC method applying muscle-related markers. Compared with untreated control sections, vimentin and desmin showed increased staining after the exposure. However, myoglobin immunostaining was not improved. Therefore, some tissue antigens or epitopes were retrieved that were masked by formalin-fixation, but the retrieval was not universal.

A gastric tumor was pointed out in 43-year-old woman as a result of a medical check-up. We performed partial gastrectomy based on a preoperative diagnosis of a gastrointestinal stromal tumor (GIST). However, as intraoperative frozen section histological examination showed lymph node metastasis, we performed a total gastrectomy simlilar to cases of gastric cancer with lymph node dissection. Postoperative histological examination showed that the tumor was positive for c-kit immunohistochemically, and consisted of spindle cells. We made a diagnosis of GIST of the stomach with lymph node metastasis. Lymph node metastasis in a case of GIST is rare, and has a poor prognosis. We report a case of a rare gastrointestinal stromal tumor with lymph node metastasis, followed up for 7 years with no evidence of recurrence after radical surgery with lymph node dissection.

A 73-year-old woman who had a history of primary biliary cirrhosis developed rapidly progressive glomerulonephritis and pulmonary-renal syndrome. She was found to have anti-mitochondrial antibody (AMA) and myeloperoxidase (MPO) anti-neutrophil cytoplasmic antibody (ANCA). She also had weak anti-glomerular basement membrane (GBM) antibody. She was treated with methylprednisolone pulse therapy, but died of respiratory failure. On postmortem examination, both lungs showed diffuse hemorrhage and the immunofluorescence study of the kidney revealed linear immunoglobulin G (IgG) deposition along the glomerular basement membrane.

Female house musk shrews (Suncus murinus, Insectivora) were given a single i.p. dose of 30 mg/kg dimethylnitrosamine (DMN) at 8 weeks of age which was lethal 36 to 48 hrs after dosing. Liver tissues were collected from shrews killed 3, 6, 16, 24 and 36 hrs after treatment, and the sequential development of the lesions was characterized. DMN induced acute centrilobular cell injury. In 6 hrs, a few cells became apoptotic in the centrilobular area; the number increased at 16 hrs and 24 hrs, and was prominent at 36 hrs. There was no inflammatory reaction or necrosis and hemorrhage was not obvious. These apoptotic cells as well as normal appearing cells in the centrilobular area were labeled by the TUNEL method. In both hepatocytes and endothelial cells, apoptosis was confirmed electron microscopically as nuclear chromatin condensation at the periphery with no mitochondria swelling. When an i.p. dose of 10 mg/kg DMN was given twice at 8 and 9 weeks of age, no acute toxicity was induced, and the liver of shrews surviving for 50 weeks of age was normal with no tumor formation. These findings indicate that a single i.p. administration of 30 mg/kg DMN induced severe and fatal toxicity on liver tissues in shrews due to apoptosis, whereas 2 x 10 mg/kg DMN had no carcinogenic effect.

Immunohistochemical detection of estrogen and progesterone receptors (ER and PgR, respectively) was performed in 67 cases of Japanese female invasive breast carcinoma on methacarn-fixed paraffin-embedded sections using monoclonal antibodies against ER (1D5) and PgR (10A9) with an antigen-retrieval technique based on microwave exposure in citrate buffer solution. Staining localized in nuclei and specimens containing >=20% specificially stained tumor cell nuclei were considered ER- or PgR-positive; the positive rate was 37% (25/67) for ER and 45% (30/67) for PgR. The immunohistochemical (IHC) results were compared with cytosolic receptors obtained from tissue homogenates as measured by dextran-coated charcoal (DCC) assay, in which tumors were considered ER- and/or PgR-positive when receptor concentration was greater than 3 or 5 fmol/mg cytosolic protein, respectively, and agreement between the two methods was 71.6% for ER and 80.6% for PgR. The sensitivity and specificity were 53.5 and 91.7% for ER-IHC, and 77.4 and 83.3% for PgR-IHC, respectively. In relation to age of the patient and stage of the cancer, ER immunoreactivity correlated with patient's age (>=55 vs 55, P = 0.032), and proportionally increased with aging ( P = 0.0084) using the Kruskal–Wallis statistics. PgR-immunoreactivity correlated with nodal involvement ( P = 0.031) by the TNM system. However, no correlations were found between the results of the DCC assay and any of the clinical parameters examined. Thus, immunohistochemical assay may provide valuable information in predicting prognosis and response to endocrine therapy.

The expression and distribution of tenascin, an extracellular matrix glycoprotein, was investigated immunohistochemically using an anti-human tenascin monoclonal antibody (RCB 1) in formalin-fixed paraffin-embedded tissues obtained from 79 patients with skin appendage tumours, and compared with adjacent normal skin. Tissue specimens were pretreated with actinase and processed by the labelled streptavidin-biotin method. In normal skin, tenascin immunoreactivity was consistently found around the ductal portion of the sweat glands, around the lower part of the hair follicle and hair bulbs, and around or within blood vessels. Immunoreactivity was also observed variably around secretory coils of the sweat glands, and below the epidermis. No immunoreactivity was seen around the sebaceous glands. Tumours originating from sweat glands and hair follicles expressed tenascin around the tumour cells nests, while sebaceous gland tumours were immunonegative. Thus, tenascin expression in skin appendage tumours generally resembled that in corresponding normal tissue.

Background/Aims: Photodynamic therapy (PDT) has a promising effect on nonresectable cholangiocarcinoma (CC) but its long term data are not available. This study examined the long term outcome and factors associated with increased survival after PDT for hilar cholangiocarcinoma. Methods: A list of 393 patients with a diagnosis of hilar CC was retrieved from the data base of Soonchunhyang University Hospital (Seoul, Korea) from January 1, 2001, to April 1, 2010. We retrospectively reviewed the records of 74 patients who underwent PDT in addition to biliary stenting with/without chemoradiation. Results: The median overall survival from the date of diagnosis to death or the last follow-up was 11.7months (range, 2.2-78.4) respectively. After PDT on negative bile duct biopsy specimens, a complete remission was observed in 1.3% (1/74) of the patients who had superficial tumor depth without lymph node involvement. In multivariate analysis with Cox regression model, increasing the time to treatment after diagnosis was statistically significant predictors of shorter survival after PDT. [Odds ratio, 3.25; 95% confidence interval (CI), 1.90-4.71, p= 0.034] Conclusion: The early treatment of PDT after diagnosis showed the survival benefit in advanced hilar CC.

We have tried to demonstrate and identify colony-stimulating factor (CSF) activity in the plasma, pleural fluid, ascites or culture supernatant of tumor cells in 11 patients with malignant tumors associated with unexplained persistent leukocytosis. The specimens were treated with anti-granulocyte (G)-CSF or anti-granulocyte/macrophage (GM)-CSF monoclonal antibodies, then added to GM-progenitor (CFU-GM) cultures without exogenous CSFs. In all patients, untreated specimens generated CFU-GM-derived colonies, and colony formation was clearly inhibited by only one of the two antibodies, indicating the presence of either G-CSF or GM-CSF in the specimens. Furthermore, we measured the concentrations of G-CSF or GM-CSF in the specimens using an enzyme-linked immunosorbent assay, and confirmed the results by CFU-GM assay. Two patients were shown to have GM-CSF-producing tumors, while the other patients were G-CSF-producing. These assays are useful in identifying CSF activity in patients with CSF-producing tumors.

Efficacy and safety of a new injectable cephem antibiotic, cefpirome sulfate (hereafter, CPR), against respiratory tract infections were examined and compared with those of a control drug, ceftazidime (hereafter, CAZ). As a rule, CPR 0.5 g twice a day, 1.0 g twice a day, or CAZ 1.0 g twice a day (hereafter CPR 0.5 g group, CPR 1.0 g group, and CAZ group) was administered for 14 days and the following results were obtained. 1. The total number of cases was 470 (155 cases in the CPR 0.5 g group, 160 cases in the CPR 1.0 g group, and 155 cases in the CAZ group). Among them 390 cases were subjected to analyses of clinical efficacy by the efficacy evaluation committee (131 cases in the CPR 0.5 g group, 131 cases in the CPR 1.0 g group and 128 cases in the CAZ group). 2. Efficacy rates determined by the efficacy evaluation committee were 82.4% (108/131) for the CPR 0.5 g group, 81.7% (107/131) for the CPR 1.0 g group, and 83.6% (107/128) for the CAZ group. Efficacy rates determined by the physician in charge were 82.0% (105/128) for the CPR 0.5 g group, 80.5% (99/123) for the CPR 1.0 g group, and 88.5% (108/122) for the CAZ group. No statistically significant difference was observed among the 3 groups. In evaluation of equivalency, clinical efficacy for the CPR 0.5 g group and the CPR 1.0 g group determined by the clinical efficacy evaluation committee was proved to be statistically equivalent to that for the CAZ group. 3. In patients with pneumonia, efficacy rates determined by the efficacy evaluation committee were 87.1% (61/70) for the CPR 0.5 g group, 80.7% (71/88) for the CPR 1.0 g group, and 78.9% (56/71) for the CAZ group. Efficacy rates determined by the physician in charge were 85.3% (58/68) for the CPR 0.5 g group, 80.7% (67/83) for the CPR 1.0 g group, and 86.2% (56/65) for the CAZ group and no statistically significant difference was observed among the 3 groups. In patients with chronic respiratory tract infection, efficacy rates determined by the efficacy evaluation committee were 77.0% (47/61) for the CPR 0.5 g group, 83.7% (36/43) for the CPR 1.0 g group, and 89.5% (51/57) for the CAZ group. Efficacy rates determined by the physician in charge were 78.3% (47/60) for the CPR 0.5 g group, 80.0% (32/40) for the CPR 1.0 g group, and 91.2% (52/57) for the CAZ group. No statistically significant difference was observed among the 3 groups.(ABSTRACT TRUNCATED AT 400 WORDS)