Muramyl dipeptide-Lys stimulates the function of human dendritic cells.

Muramyl dipeptide (MDP)-Lys (L18), a synthetic MDP analogue derived from bacterial cell walls, has been reported to be a potent immunoadjuvant that enhances protective immunity against pathogens and tumors by stimulating immune-competent cells, such as monocytes and macrophages. However, it is not known whether MDP-Lys modulates the function of dendritic cells (DCs), which are the most potent antigen-presenting cells and play a crucial role in initiating T cell-mediated immunity. Therefore, we examined the effects of MDP-Lys on the expression of surface molecules, cytokine production, and antigen-presenting function of human DCs generated from peripheral blood cells in the presence of interleukin (IL)-4 and granulocyte-macrophage colony-stimulating factor. We found that MDP-Lys markedly up-regulated the expression of CD80, CD83, CD86, and CD40, but not human leukocyte antigen-DR, and stimulated the production of tumor necrosis factor-alpha, IL-6, IL-8, IL-10, and IL-12 (p40) by human DCs in a dose-dependent manner. Furthermore, MDP-Lys-treated DCs showed enhanced antigen-presenting function compared with untreated DCs, as assessed by an allogeneic mixed lymphocyte reaction. These results suggested that the immunoadjuvant activity of MDP-Lys in vivo is mediated, in part, by its stimulation of DC function.

Spontaneous uterine adenocarcinomas in aged rats and their relation to endocrine imbalance.

In addition to spontaneous uterine endometrial adenocarcinomas at a high incidence (35.1%), development of endometrial hyperplasia/adenoma was also frequently detected in rats of the Donryu strain. The total yield of all observed proliferative endometrial lesions was very high (60.6%). The tumors arose commonly in the uterine horn of aged rats. Histologically, most demonstrated glandular structures, consisting of cuboidal or columnar cells with weak eosinophilic or basophilic cytoplasm and large nuclei. In about half of the animals with adenocarcinomas, metastasis to remote organs such as the lung was observed. Histological examination of the ovary and vaginal epithelium revealed ovarian cysts, atrophy of the ovary and cornification of the vaginal epithelium more frequently in rats with endometrial carcinomas than in animals without tumors. These findings indicate that adenocarcinoma development in Donryu rats is associated with endocrine imbalance [increased serum estrogen: progesterone (E2:P)ratios]. By comparative investigation of strain differences, it was confirmed that irregular estrous cycles began earlier with higher incidence in Donryu rats than in F344 rats, a low-incidence strain. Histological findings of the ovary and vaginal epithelium also suggested relatively increased estrogen levels in Donryu rats compared to F344 rats. Estimated plasma values of gonad steroids showed that the E2:P ratio in Donryu rats at 12 months of age was about five times that in F344 rats. These results therefore indicate that hormone imbalance, particularly an increased E2:P ratio, may play an important role in the spontaneous occurrence of endometrial adenocarcinoma in Donryu rats.

Two-year carcinogenicity study of 6-mercaptopurine in F344 rats.

The carcinogenicity of 6-mercaptopurine (6-MP), an anticancer drug, was examined in F344 rats of both sexes, administered the chemical at dietary levels of 0 (control), 25 ppm or 50 ppm for 2 years. Many tumors developed in all groups including the control group, the organ distribution and histological types being similar to those reported for spontaneous lesions. In males, there was no significant increase in the incidence of any tumor in the treated groups over that in the control group. In females, however, positive trends were noted in the occurrence of C-cell tumors, pheochromocytomas, uterine adenocarcinomas and gliomas, and the incidences of C-cell tumors and pheochromocytomas in the 50 ppm group were significantly higher than the values in the respective control group. In addition, the total numbers of malignant tumors increased significantly in the female 50 ppm group. However, most of the tumors demonstrating increase are frequently observed spontaneous lesions in this strain of rats, and their incidences in the present female control group were lower than in our historical data. In addition, there were no significant differences in the incidences of preneoplastic changes and induction times for the above-listed tumors between the female control and the 50 ppm groups. These results thus indicated that while the carcinogenic potential of 6-MP can not be precluded, it can be only very weak or marginal, after continuous administration in the diet at the 50 ppm level for 2 years. The leukemogenic action of 6-MP was negative under the present experimental conditions.

Carcinogenicity and organ specificity of N-trimethylsilylmethyl-N-nitrosourea (TMS-MNU), N-neopentyl-N-nitrosourea (neoPNU), and N-methyl-N-nitrosourea (MNU) in rats.

The carcinogenicity and organ specificity of TMS-MNU and neoPNU, a carbon-analogue of TMS-MNU, in rats were investigated and compared with those of MNU. Compounds were dissolved in olive oil and rats in the experimental groups received 20 weekly intragastric intubations of 10 mg/kg of MNU or equimolar amounts of TMS-MNU or neoPNU in the same manner. The experiment was terminated when the survivors were sacrificed at the 52nd week after the final administration. In the TMS-MNU and MNU groups, tumors of the forestomach were induced and the incidence was 100% in the groups of both sexes. In addition, tumors of the glandular stomach, nervous system, kidney, and lung were also observed in these groups. Neurogenic tumors were found more frequently in the MNU group than in the TMS-MNU group. The incidence of lung tumors, however, was higher in the TMS-MNU group than in the MNU group. On the other hand, in the control and neoPNU groups, no tumor was found in these organs except the lung, and all tumors observed in these two groups were histologically similar to spontaneous ones in this strain of rats. These results indicate that the carcinogenicity of N-alkyl-N-nitrosoureas is dependent on the chemical structure of their alkyl chain. The result of the present study coincides with the previous result that the species of TMS-MNU in the alkylating step is the same as that of MNU, but different from neoPNU. The difference in the organ specificity between TMS-MNU and MNU demonstrates that the organ specificity is dominantly dependent on the distribution of the chemicals, since TMS-MNU may possibly be distributed differently from MNU because of its different partition property.

Lack of toxicity/carcinogenicity of monosodium succinate in F344 rats.

The toxicity/carcinogenicity of monosodium succinate, a food additive, was examined in F344 rats. The oral LD50 was greater than 8 g/kg body weight. In a 13-wk subchronic oral toxicity study, the only toxicological finding was suppression of body-weight gain in groups given greater than or equal to 2.5% monosodium succinate in the drinking-water. Histological examination revealed no toxic lesions specifically caused by the compound in any organs of any of the treated rats. The maximum tolerated dose was determined to be 2-2.5% on the basis of body-weight depression. In a long-term (2-yr) toxicity/carcinogenicity study, monosodium succinate was given ad lib. in drinking-water (distilled water) at levels of 0, 1 or 2% to groups of 50 male and 50 female rats. No toxic lesion specifically caused by long-term administration of monosodium succinate was detected. No dose-related increase was found in the incidences of tumours in any organ or tissue except for C-cell tumours of the thyroid gland of females. The incidence of these tumours in females given the 2% dose was higher than that in controls but not significantly so, and a positive trend for this tumour was noted in females. C-Cell tumour is one of the most commonly observed spontaneous tumours in ageing female rats of this strain and occurs at a variable incidence. There was no difference between the female control and treated groups in the incidence of preneoplastic change of the thyroid gland. Furthermore, the incidence of C-cell tumours in the female control group was lower than that in our historical controls. It is concluded that the increase in C-cell tumours in the female high-dose group and the detection of a positive trend for this tumour in females were probably a function of experimental variability and were not related to treatment. The results indicate that monosodium succinate had neither toxic nor carcinogenic activity in F344 rats when it was given continuously at levels of 1 or 2% in the drinking-water for 2 yr.

Diffuse panbronchiolitis and rheumatoid arthritis-associated bronchiolar disease: similarities and differences.

There is a considerable overlap between diffuse panbronchiolitis (DPB) and bronchiolar disease associated with rheumatoid arthritis. The present study assessed how these conditions could be differentiated. The subjects included 25 DPB patients and 15 RA patients with bronchiolar disease (RA-BD). Patients with either condition had chronic cough, purulent sputum, dyspnea and coarse crackles. Most patients with either DPB or RA-BD had a history of sinusitis as well as elevated cold hemagglutin titers and decreased levels in partial pressure of oxygen (PaO2), forced expiratory volume in one second (FEV1.0) and V 25/Ht. On histological examination, both conditions also shared various histological patterns although panbronchiolitis lesions were more common in DPB than RA-BD (68% vs 20%) and bronchiolar obliteration appeared to occur at more proximal sites in RA-BD than DPB. However, there were important differences: long-term treatment with erythromycin had less effect in RA-BD than DPB, and the frequency of HLA B54 tended to be higher in DPB than RA-BD (50.0% vs 22.2%), suggesting that they are distinct conditions.

Teratoma of the pituitary gland in a young male rat.

A pituitary teratoma was found in a 5-week-old (37-day-old) male Donryu rat. The tumour (10 x 11 x 9 mm) was round in shape and white in colour. The cut surface was solid without haemorrhagic or cystic change. Histologically, it was composed of various kinds of tissue components including mature and immature elements derived from the three embryonic germ layers, i.e., nervous tissue, cartilage, bone, squamous epithelial element, glandular element lined by one or more layers of ciliated columnar or cuboidal epithelial cells, striated muscle tissue, adipose tissue and connective tissue. Neuroepithelial rosettes and immature or embryonal epithelium as well as immature cartilage were intermingled with mature somatic tissues.

[A 13-week subacute oral toxicity study of 6-amidino-2-naphthyl 4-[(4,5-dihydro-1H-imidazol-2-yl) amino] benzoate dimethanesulfonate (FUT-187) in rats].

The toxicity of FUT-187, a synthetic protease inhibitor, was investigated in Sprague-Dawley rats. FUT-187 was given orally to the rats at doses of 2, 10, 50, 250 and 1250 mg/kg/day for 13 weeks, then the drug was withdrawn for 5 weeks for recovery. The results are summarized as follows: In the 1250 mg/kg/day group, 9 out of 20 males died with decreased body weight and exhaustion. Histopathological examination revealed renal papillary necrosis, ulcer in the urinary bladder, hemostatic lesions in the lungs and liver, ulcer or erosion in the stomach, duodenum and jejunum. The surviving animals in this group showed swelling of the limbs due to synovitis, transient salivation immediately after administration, suppression of growth with decreased food consumption. Urinalysis revealed a low pH, increased ketones and bilirubin excretion, dark yellowish change in color, the appearance of "leaflet-shaped" crystals and increased red blood cells and epithelial cells in the urinary sediment, increased water intake, decreased specific gravity and decreased sodium, potassium and chloride in the urine. Hematologically, there was an increase in the white blood cell count. A biochemical analysis of the blood revealed decreased amylase activity, glucose and total protein levels and increased GOT activity and inorganic phosphorus levels. Pathological changes were observed in the pancreas, kidney, digestive tract, urinary bladder and liver. The pancreas showed macroscopical enlargement and increased organ weight. Histopathologically, there were several alterations in the acinar cells, such as vacuolization due to increased fat droplets, nuclear irregularity, prominent nucleoli, irregular arrangement and vesiculation of rough endoplasmic reticulum (rER), dilatation of developed Golgi apparatus and increased free ribosomes. In the kidney, increased weight and pigmentation in the proximal tubular epithelium were noted. Electron microscopically, these pigments were recognized as secondary lysosomes containing filamentous material and electron dense granules within a lucent matrix. In the digestive tract, ulcer or erosion in the stomach and duodenum, and villous proliferation in the small intestine were observed. Furthermore, hyperplasia and vacuolization were noted in the mucosal epithelium of the urinary bladder. In addition, loss of perilobular fat droplets in the liver and increased adrenal weight without histological change were observed. After a 5-week recovery period, these changes disappeared almost completely. In the 250 mg/kg/day group, slight suppression of growth the appearance of "leaflet-shaped" crystals in the urinary , sediment, increased water intake and decreased sodium in the urine were observed. The pancreas showed enlargement, increased weight, acinar cell hypertrophy with increased zymogen granules, fine vacuolization, slight derangement and vesicular of rER, and dilatation of Golgi apparatus.(ABSTRACT TRUNCATED AT 400 WORDS)

[Pulmonary cryptococcosis exhibiting diffuse multiple nodular shadows].

We report a case of pulmonary cryptococcosis showing diffuse multiple nodular shadows in all lung fields. A 39-year-old woman with no immunological abnormalities was admitted with complaints of cough and sputum. She had experienced measles 4 weeks prior to admission. Chest x-ray films revealed diffuse nodular opacities throughout the lung fields, a finding suggestive of metastatic lung cancer. Detailed examinations, including transbronchial lung biopsy, were not conclusive. A diagnosis of pulmonary cryptococcosis was made on the basis of findings from video-assisted thoracoscopic biopsy. Primary pulmonary cryptococcosis usually appears as a solitary nodule or limited infiltration. Immunologically compromised hosts commonly demonstrate various abnormal shadows, such as the multiple nodular shadows observed in our patient. It has been reported that measles infection can cause temporary immune suppression. Secondary immunodeficiency resulting from the preceding infection with measles could explain the unusual chest x-ray findings in this case.

[Pyothorax associated lymphoma treated by chemotherapy after thoracostomy].

A 62-year-old man with a medical history that included artificial pneumothorax therapy at the age of 18 was admitted to our hospital because of persistent cough. Chest computed tomographic scans disclosed atelectasis in the right lung and pyothorax surrounded by calcifications. Radiographic examination failed to disclose any tumors. After admission, high grade fever developed due to aggravated pyothorax infection. Because antibiotic therapy and drainage failed, open window thoracostomy was performed. Tumors were found along the wall of the pyothorax cavity, and examination of resected specimens yielded a diagnosis of non-Hodgkin's lymphoma, diffuse large cell type (B-cell lineage). It was difficult to close the pyothorax cavity due to infection and lymphoma. Therefore, with the thoracic window open, the patient was given combination chemotherapy including CHOP (6 courses) and DeVIC (7 courses). He died of disseminated intravascular coagulation 17 months after thoracostomy. In patients with pyothorax associated lymphoma, chemotherapy is sometimes difficult to perform because of persistent pyothorax infection. Although edema and ascites due to protein loss from the tumor complicated the treatment of our patient, we concluded that open window thoracostomy is effective in managing pyothorax prior to and during chemotherapy.

[Interstitial pneumonia preceding systemic sclerosis].

We describe a 57-year-old man with interstitial pneumonia associated with systemic sclerosis. About 3 years prior to the appearance of distinctive signs of systemic sclerosis, he was admitted to our hospital with a chronic dry cough. A chest roentgenogram on admission revealed reticulonodular shadows in both lung fields. There were no abnormal laboratory findings. Open lung biopsy specimens revealed patterns indicative of usual interstitial pneumonia, and myxomatous connective tissue within the lumen of the airways. Skin biopsy specimens showed heightened levels of collagen in the dermis, a finding consistent with systemic sclerosis. The patient was given a diagnosis of lung involvement preceding systemic sclerosis despite the absence of concurrent skin symptoms.

[A case of pulmonary alveolar proteinosis presenting with peripheral infiltrates].

We report a case of pulmonary alveolar proteinosis (PAP). A 39-year-old asymptomatic woman was admitted to our hospital because of abnormal shadows on chest X-ray films. Chest X-ray films revealed peripheral infiltrates in both lungs. Computed tomographic examination showed patchy peripheral ground-glass attenuation, concentrated subpleurally. Bronchoalveolar lavage fluid was clear. Because transbronchial lung biopsy findings were inconclusive, a VATS-biopsy was performed. The specimens demonstrated accumulation of proteinaceous materials within alveolar spaces. The patient was given a diagnosis of PAP. Although the distribution of radiographic shadows varies in patients with PAP, perihilar or centralized shadows usually predominate. In our patient, subpleural areas of the lung were affected almost exclusively.

[Recurrent pulmonary infarction associated with familial protein S deficiency type III].

A 38-year-old woman was admitted to our hospital because of recurrent chest pain and fever. Chest X-ray films and computed tomograms showed subpleural consolidation containing small cavity-like opacities. Open lung biopsy revealed non-infectious abscess and vessels with organizing thrombus. The patient was given a diagnosis of pulmonary infarction due to the existence of deep venous thrombosis. Coagulation studies demonstrated that she had decreased plasma protein S activity, whereas her free and total protein S antigen levels were normal. Because her mother and maternal uncle and aunt also demonstrated decreased protein S activity with normal plasma protein S antigen levels, the patient was considered to be affected by familial protein S deficiency type III.

[A comparative study of the prognosis for Japanese patients with idiopathic interstitial pneumonia or BOOP based on histopathologic subsets].

We explored the prognosis for 123 patients with either idiopathic interstitial pneumonia (IIP) or bronchiolitis obliterans organizing pneumonia (BOOP). All patients underwent either open lung biopsy or thoracoscopic lung biopsy procedures. The histopathologic diagnosis of IIP included patients with usual interstitial pneumonia (UIP), nonspecific interstitial pneumonia (NSIP), and desquamative interstitial pneumonia with respiratory bronchiolitis-associated interstitial lung disease. The prognosis was poorest for patients with a histologic diagnosis of UIP, and excellent for those who received a diagnosis of BOOP. Although the prognosis is generally considered to be good for patients with NSIP, some NSIP patients in our study died. Histopathologic diagnosis based on surgical lung biopsy is useful in evaluating the prognosis for patients with IIP.

[A 28-day repeated dose toxicity study of nitrobenzene in F344 rats].

To examine toxicities of nitrobenzene as part of the re-evaluation of toxicities of existing chemicals, a 28-day repeat dose toxicity study was performed in male and female F344 rats at dosages of 0, 5, 25 and 125 mg/kg/day of nitrobenzene. All rats in each group consisting of 6 males and 6 females received a daily intragastric administration of this chemical for 28 days. Additional two groups of animals exposed to 0 and 125 mg/kg/day were used for examinations of subsequent recovery for 2 weeks. One female in the 125 mg/kg group died on day 27. Decreased movement, pale skin, gait abnormality and decreases of body weights or their gains were seen in the 125 mg/kg group. Hematology revealed decreases of RBC, Hb and Ht in the 25 and/or 125 mg/kg groups. Blood biochemistry revealed increases of total cholesterol and albumin and decreases of BUN in the 25 and 125 mg/kg groups, and increases of A/G ratio in both sexes and ALT, ALP and total protein in females in the 125 mg/kg group. In the organ weight, increases of the liver, spleen, kidney weight and decreases of the testis and thymus were seen in the 125 mg/kg group. In addition, the increased liver weight was also seen in males receiving 5 mg/kg, and the increased spleen weight in both sexes receiving 25 mg/kg. Histopathology revealed spongiotic changes and brown pigmentation in perivascular region of the cerebellum, increased extramedullary hematopoiesis of the liver, brown pigmentation of renal tubular epithelium and degeneration of seminiferous tubular epithelium and atrophy of seminiferous tubule in the 125 mg/kg group, and congestion, increased brown pigmentation in red pulp and increased extramedullary hematopoiesis of the spleen and increased hematopoiesis of the bone marrow in treated groups. Findings mentioned above disappeared or tended to be decreased during or at the end of the recovery period. Although no effect-dose level was detected in this study, severe anemia and disorder of spermatogenesis and central nervous system which have been reported in the long-term toxicity study could be reconfirmed.

Ultrastructural study of antral G cells in patients with duodenal ulcer: effect of Helicobacter pylori eradication.

BACKGROUND: It is established now that Helicobacter pylori infection is associated with exaggerated gastrin release to meals and other stimuli and that the abnormal secretion of gastrin is reversed after successful treatment of the infection. By comparing morphology of G cells from the same patients before and after treatment, we were able to investigate the ultrastructural effect of cure of H. pylori on G cells. METHODS: Gastric mucosal biopsy specimens were obtained from 10 patients with duodenal ulcer before and 3, 6, and 9 months after cure of H. pylori infection. Negative controls consisted of four healthy volunteers without H. pylori infection. G cells were evaluated by immunohistochemical and electron microscopy. Treatment with H2-antagonists was continued for 6 months after cure of the H. pylori infection. RESULTS: Ultrastructural studies of secretory granules of antral G cells in controls displayed a broad range of electron density ranging from dark with a full appearance to totally electron-lucent with a "vacuolating" appearance. In duodenal ulcer patients before treatment, electron-lucent vacuolating granules predominated. After elimination of H. pylori, G-cell granules showed a marked increase in electron density close to that of controls. CONCLUSIONS: Our study showed that the density of granules of G cells is decreased in H. pylori-infected duodenal ulcer patients compared to that in H. pylori-negative controls and is consistent with enhanced gastrin release. Cure of H. pylori infection was associated with return to normal density of G-cell granules.

[Subchronic oral toxicity study of calcium lactate in F344 rats].

A subchronic toxicity study of calcium lactate was carried out in male and female F344 rats to estimate the maximum tolerated dose for a subsequent long-term toxicity/carcinogenicity study. Experiment I: Rats were divided into 6 groups, each consisting of 5 males and 5 females. Calcium lactate was dissolved in water at concentrations of 5, 2.5, 1.25, 0.6, 0.3 and 0%, each animal group was given one of these solutions as the drinking water for 13 wk. In all groups, basic diet (CRF-1) was given ad libitum. No fatalities occurred. In all treated groups, including the 5% group, a less than 10% depression of body-weight gain as compared with the control group was observed. Some parameters in the hematological and biochemical data demonstrated change in the treated groups. On histological examination, however, no severe toxicological findings were found in any of the treated groups. Experiment II: Rats were fed synthetic diet B, containing 30, 20, 10, 5 or 0% calcium lactate. In the highest dose group, body weight-gain was strongly reduced as compared with the control group. Histological examination revealed nephrocalcinosis in all groups, including the control group, and adverse dose-effect relation was observed with regard to degree of its development. Females exhibited this lesion to a greater extent than males. Experiment III: Rats were given CRF-1 or synthetic diet B for 8 wk. Nephrocalcinosis was found only in the group given synthetic diet. It was ascertained that the nephrocalcinosis observed in Exp. II and III was dependent on the low Ca/P ratio (Ca/P: less than 1) of the synthetic diet B.(ABSTRACT TRUNCATED AT 250 WORDS)

[Subchronic oral toxicity study of tannic acid in F344 rats].

A 13-week subchronic oral toxicity study of tannic acid (TA) was carried out in F344 rats at dose levels of 0, 0.025, 0.05, 0.1, 0.2 and 0.4% in the drinking water, to determine appropriate dose levels for a subsequent 2-year carcinogenicity study. The rats were randomly allocated to 6 groups, each consisting of 12 males and 12 females. No animals died during the administration period. There were no significant difference in body weight gain, food consumption and organ weights between the treated and control groups, although a slight decrease in water intake was seen in the 0.4% TA treated group. No specific changes were observed in any parameters in the hematological and biochemical investigations. Histopathological examination, revealed toxic changes in the TA treated male groups, in the form of necrosis in the liver, but toxicologically it was of minor importance. From these results, it was concluded that the provable maximum tolerable dose of TA in the drinking water would be more than 0.4%. In consideration of the avoidance of drinking water, the maximum tolerable dose of tannic acid was determined to be 0.5%, when given in the drinking water.