Young C3H mice infected with Toxoplasma gondii are a novel experimental model of communicating hydrocephalus.

The effects of leptomeningeal inflammation on the development of hydrocephalus are less understood than those of obstructing the flow of cerebrospinal fluid (CSF) in animal models. We succeeded in introducing a novel experimental model of hydrocephalus and analysed changes in histopathology and CSF flow in mice infected with an avirulent Fukaya strain of Toxoplasma gondii (T. gondii). Six to 7 week-old male mice were orally inoculated with a brain homogenate containing ten T. gondii cysts. The cerebral ventricles became enlarged in all C3H/HeN and C57BL/6 mice 4 weeks after T. gondii infection, but mildly in BALB/c mice. In addition to the lateral ventricle, the third and fourth ventricles and Sylvian aqueducts were dilated in all mice. Lymphocytes and monocytes infiltrated the subarachnoid space. Indian ink particles required more time to pass from the lateral ventricle to the cervical lymph nodes, although they reached the subarachnoid space. Computed tomography ventriculography demonstrated that the CSF was not obstructed during passage through the ventricular systems, but contrast remained static in the lateral ventricle only in infected mice. These results indicated that the infected mice developed communicating type hydrocephalus without obstructive or mass lesions in the ventricles. The hydrocephalus that arises in mice infected with T. gondii is considered a consequence of leptomeningeal inflammation that blocks CSF circulation at the subarachnoid space, implying that leptomeningeal inflammation is important in other types of hydrocephalus.

Hedgehog signaling enhances core-binding factor a1 and receptor activator of nuclear factor-kappaB ligand (RANKL) gene expression in chondrocytes.

Hedgehog signaling is considered to play a crucial role in chondrogenesis by regulation through a network of cytokine actions, which is not fully understood. We examined the effect of hedgehog signaling on the expression of core-binding factor a1 (Cbfa1), a critical transcription factor for the development of bone and cartilage. Primary chondrocytes prepared from the costal cartilage of newborn mice were treated with N-terminal fragment of recombinant murine sonic hedgehog (rmShh-N). Northern blot analysis indicated that Cbfa1 mRNA expression levels in the chondrocyte cultures were elevated by the treatment with rmShh-N. rmShh-N treatment enhanced 1.8 kb Cbfa1 promoter activity in chondrocytes, suggesting the presence of transcriptional control. As Cbfa1-binding site(s) have been located in the promoter of the receptor activator of nuclear factor-kappaB (RANK) ligand (RANKL) gene, we also examined RANKL expression. rmShh-N treatment upregulated RANKL and RANK mRNA expression levels in chondrocytes. Interestingly, RANKL suppressed the hedgehog enhancement of alkaline phosphatase activity in chondrocytes, suggesting the presence of a link between these signaling molecules. We conclude that hedgehog signaling activates Cbfa1 gene expression through its promoter in chondrocytes, and also activates and interacts with RANKL to maintain cartilage development.

Mechanism of hypergammaglobulinemia by HIV infection: circulating memory B-cell reduction with plasmacytosis.

The mechanism of hypergammaglobulinemia in patients infected with HIV has remained unclear in spite of the identification of a reduction of CD4+ T cells. The amounts of CD27+ memory B cells were remarkably reduced in the peripheral blood and immunoglobulin (Ig) production was diminished in HIV-infected patients. Some of the freshly isolated patients' T cells expressed the CD70 (CD27 ligand) on the surface and the CD70 expression on both of the CD4+ and CD8+ T cells was greatly enhanced by various stimuli. It was also striking that plasmacytosis was observed in patients' bone marrow. Thus, our findings suggest that CD70 expressed spontaneously or by activation on T cells of HIV-infected patients stimulates memory B cells via CD27 and promotes their differentiation into plasma cells, resulting in the elevation of serum Ig levels and the elimination of circulating memory B cells in HIV-infected patients.

[Follicular bronchiolitis associated with pulmonary tuberculosis--report of a case].

The patient, a 39-year-old man, presented in May 1997 with an untreated persistent cough with excessive sputum of 5 years' duration. He was admitted to hospital because of the severity of the symptoms and the presence of acid-fast bacilli in his sputum. These bacilli were identified as Mycobacterium tuberculosis by the polymerase chain reaction method. After treatment with antituberculous drugs was initiated, his symptoms and the patchy infiltrative shadows on his chest radiographs gradually resolved. However, the patient continued to expectorate a purulent sputum, showed diffuse micronodular shadows on chest radiographs, and had hypoxemia as well as mixed ventilatory and small airway disturbances on pulmonary function tests. We performed a video-assisted thoracoscopic biopsy of the lung, which revealed follicular bronchiolitis.

A pivotal role of cell-bound but not soluble CD4 molecules in full development of lupus-like manifestations in MRL-Fas(lprcg)/Fas(lprcg) mice.

The role of CD4 molecules in the autoimmune and lymphoproliferative syndrome caused by murine Fas mutations was studied using the novel systemic lupus erythematosus (SLE) model, MRL-Fas(lpr(cg))/Fas(lprcg) (MRL-lpr(cg)) mice, in combination with the novel mutant CD4 gene producing soluble CD4 (sCD4) instead of membrane-bound CD4 (mCD4). For this purpose, various autoimmune manifestations were compared among MRL-lpr(cg) mice homozygous (CD4slprcg), heterozygous (CD4s/mlpr(cg)), and wild-type (CD4mlpr(cg)) for the CD4 mutation. The mortality, glomerulonephritis, proteinuria, and lymphadenopathy were significantly ameliorated in CD4slprcg compared with CD4mlpr(cg) and CD4s/mlpr(cg) mice, both being comparable in these clinical characteristics. In parallel with the clinical improvement, the serum levels of immunoglobulin, anti-DNA antibodies, anti-nuclear antibodies and immune complexes, and the extent of glomerular immune deposition, were significantly lower in the former. The results indicate that mCD4 is important and can not be replaced by sCD4 in full development of SLE-like manifestations, and suggest that CD4+ T cells may aggravate the autoimmune disease by stimulating autoreactive B cells to produce autoantibodies through their helper activity in Fas mutant models. The sCD4 levels in the serum and spleen elevated with the increased accumulation of B220+CD4-CD8- (double-negative (DN)) T cells in CD4slpr(cg) mice. This, together with the significantly milder lymphadenopathy associated with lower DN T cell contents in CD4slpr(cg) than CD4mlpr(cg) mice, implies that some of abnormal DN T cells may be derived from cells of the CD4 lineage.

[Evaluation of CT appearance of Mycobacterium avium complex infection--comparison with bronchiectasia].

We compared computed tomographic pictures (CT) of primary pulmonary mycobacterium avium complex (MAC) infection with bronchiectasia (BE). These patients were examined during the period from 1988 to 1996. There were 51 patients of pulmonary MAC infection group (13 males and 38 females) with a mean age of 67.5 years and 27 patients of BE group (8 males and 19 females) with a mean age of 53.9 years. The results were as follows: 1. In both groups, bronchiectasis was most commonly found in the middle lobe and in the lingula. In pulmonary MAC infection group, 38 patients (74.5%) had bronchiectasis in the middle lobe, and 40 patients (78.4%) had them in the lingula. In BE group, 18 patients (66.7%) had them in the middle lobe and in the lingula respectively. In the lower lobe, bronchiectasis of BE group was observed more frequently. And in pulmonary MAC infection group, 15 patients (29.4%) had bronchiectasis in the central zone, 42 patients (82.4%) had them in the intermediate zone, and all 51 patients had them in the peripheral zone. On the other hand, in BE group, 18 patients (66.7%), 27 patients (100%) and 21 patients (77.8%) had them in the central zone, in the intermediate zone and in the peripheral zone respectively. Bronchiectasis of pulmonary MAC infection was observed in peripheral zone more frequently than that of BE group. 2. By the type of bronchiectasis, saccular bronchiectasis was observed more frequently in BE group than in pulmonary MAC infection group. 3. Nodules were seen in pulmonary MAC infection group significantly more than in BE group. They were seen in 96% of lobes with bronchiectasis, and 50% of lobes without bronchiectasis in pulmonary MAC infection group. It was thought that the bronchiectasis with nodules predominantly in the peripheral zone was a characteristic CT finding of primary pulmonary MAC infection. As this finding was different from that of BE, it is suggested that primary pulmonary MAC infection caused this type of bronchiectasis.

CD27/CD70 interaction augments IgE secretion by promoting the differentiation of memory B cells into plasma cells.

The induction of IgE switching in B cells requires several signals given by cytokines and cell contact-delivered signals. Here, we investigated the role of CD27/CD70 interaction in B cell IgE synthesis. The addition of CD27 ligand (CD70) transfectants to B cell cultures increased the IgE synthesis synergistically in the presence of IL-4 plus anti-CD40 mAb (anti-CD40). The effect of CD70 transfectants was dose dependent and was completely blocked by anti-CD70 mAb. CD27+ B cells had the ability to produce IgE, which was increased by contact with CD70 transfectants, whereas CD27- B cells did not produce IgE. CD27/CD70 interaction enhanced B cell proliferation in the presence of IL-4 or IL-4 plus anti-CD40. The augmentation of B cell proliferation by CD70 transfectants was apparent in CD27+ B cells, but was mild in CD27- B cells. The helper activity for IgE synthesis by the CD27/CD70 interaction did not contribute to the enhancement of germline epsilon transcripts. Flow cytometric and morphological analyses demonstrated that the addition of CD70 transfectants to B cell cultures remarkably promoted differentiation into plasma cells in the presence of IL-4 and CD40 signaling. Finally, CD27 cross-linking resulted in the up-regulation of positive regulatory domain I-binding factor-1. Taken together, our findings indicate that signaling via CD27 on B cells induces IgE synthesis, in cooperation with IL-4 and CD40 signaling, by promoting the generation of plasma cells through up-regulation of positive regulatory domain I-binding factor-1.

[A study of sialylated carbohydrate antigen in patients with benign bronchopulmonary disease].

We studied the levels of carbohydrate antigen (CA 19-9, SLX, CA 50, Span-1, and Dupan-2) in serum, bronchoalveolar lavage fluid, and tissue from patients with benign bronchopulmonary disease. Patients had bronchiectasis, healed pulmonary tuberculosis, pulmonary fibrosis, or other diseases. Bronchoalveolar lavage fluid levels and immunohistochemical findings for lung tissue samples, in the absence of digestive and other diseases, suggested that elevated serum sialylated Lewis(A) (CA 19-9, CA 50, and Span-1) and Lewis(X) (SLX) antigen in patients with benign broncho-pulmonary disease are due to marked production of sialylated carbohydrate antigen in respiratory bronchioles. Common features of patients with benign bronchopulmonary disease include elevated serum carbohydrate antigen levels and bronchiectasis.

Eosinophilia, parasite burden and lung damage in Toxocara canis infection in C57Bl/6 mice genetically deficient in IL-5.

C57Bl/6 mice genetically deficient in interleukin (IL)-5 (IL-5-/-) and mice with the normal IL-5 gene (IL-5+/+) were infected with embryonated eggs of Toxocara canis. IL-5+/+ mice developed a marked eosinophilia in their peripheral bloods and bone marrows after infection. In contrast, the number of eosinophils at these sites actually decreased during the acute phase of infection in IL-5-/- mice. A smaller number of eosinophils infiltrated the lung, liver, heart and skeletal muscle of infected IL-5-/- mice than those of infected IL-5+/+ mice. Eosinophils were not produced in cultures of bone marrow cells from either IL-5+/+ or IL-5-/- mice which were stimulated with excretory secretory antigen of T. canis larvae. The capacity of cells from the bone marrow to differentiate into eosinophils when stimulated in vitro with recombinant murine IL-5 was the same whether the cells were from IL-5+/+ or IL-5-/- mice. Taken together, these results show that an IL-5-like molecule is not produced by the T. canis larvae and that IL-5 produced by host cells is solely responsible for the eosinophilia in mice infected with this nematode. The number and location of T. canis larvae were not altered in the absence of IL-5. In contrast, lung damage in infected IL-5-/- mice was less extensive than that in infected IL-5+/+ mice, although structures resembling Charcot-Leyden crystals were seen in the lungs of both IL-5+/+ and IL-5-/- mice. These results suggest that eosinophils play a role in the pathology in mice infected with T. canis.

[Tracheobronchopathia osteoplastica with "rock garden" findings on bronchoscopy].

We encountered a 65-year-old man with tracheobronchopathia osteoplastica with "rock garden" findings on fiberoptic bronchoscopy. Many nodular elevated lesions were seen on all sides of the trachea and main bronchi except the membranous portion. The diagnosis was confirmed by examination of a biopsy specimen.

[Clinical efficacy of imipenem/cilastatin sodium for respiratory infections in patients with lung cancer].

Imipenem/cilastatin sodium (IPM/CS) was administered to 102 patients with respiratory tract infections and lung cancer. Patients with other serious diseases were excluded and a total of 73 patients were enrolled. They were divided into 12 patients who underwent surgery (operated group) and 61 who did not (non-operated group); the latter group included 28 patients treated with anticancer agents or radiation therapy (treated group) and 33 untreated patients (untreated group). IPM/CS was effective in 75% of the patients, both with and without surgery. The drug was effective in 81% of the treated group, although many of the patients had Stage III or more advanced cancer, as well as bronchial occlusion. IPM/CS was also effective in 69% of the untreated group, although many of the patients have serious infections and a PS (Performance Status) of 3 or greater. Thus, IPM/CS treatment achieved good results. Bacteriological studies showed that 3 out of 4 strains in the operated group and 16 out of 18 in the non-operated group were eliminated. Safety was evaluated in all patients. Two patients (2%) experienced side effects and two others (2%) showed abnormal clinical findings, but the symptoms were mild and resolved after discontinuation or completion of therapy. In conclusion, IPM/CS was very effective for treating respiratory infections in patients with lung cancer.

A highly sensitive and selective method for the determination of Leukotriene B4 in human plasma by negative ion chemical ionization/gas chromatography/tandem mass spectrometry.

We have developed a highly sensitive and highly selective method for the determination of Leukotriene B4 (LTB4) in human plasma using negative ion chemical ionization/gas chromatography/tandem mass spectrometry (NICI/GS/MS/MS) analysis. The developed method was summarized as follows. Deuterated LTB4 (d4-LTB4) was added to human plasma samples as an internal standard, and samples were extracted by a Sep-pak C18 column. Extracted LTB4 was derivatized into the pentafluorobenzyl ester of bis-trimethylsilyl ether (PFB-TMS-LTB4) and quantified on the basis of selected reaction monitoring (SRM) at m/z 299 of [M-PFB-2TMSOH]- by NICI/GC/MS/MS analysis, which was the product ion of [M-PFB]-. The detection limit for the quantification of LTB4 in human plasma was 10 pg ml-1, sufficiently sensitive to determine the concentrations of endogenous LTB4 in human plasma. The plasma level of LTB4 measured in healthy male volunteers was 33.85 +/- 33.91 pg ml-1 (mean +/- S.D. in six volunteers). The technique of MS/MS used in this method offers much greater sensitivity and selectivity than single-stage mass spectrometry. The developed method showed good reproducibility with a simple and rapid extraction procedure, and would be useful for examining the relationship between various disease states and the levels of LTB4 in biological fluids.

Synergism of IL-3, IL-5, and GM-CSF on eosinophil differentiation and its application for an assay of murine IL-5 as an eosinophil differentiation factor.

In order to elucidate the mechanisms of eosinophil differentiation, we examined the effects of combinations of interleukin 5 (IL-5) with IL-3 and granulocyte-macrophage colony-stimulating factor (GM-CSF) on in vitro differentiation into eosinophils from bone marrow cells of ICR mice. When the amount of added IL-5 was kept constant, IL-3 exhibited dose-dependent production of eosinophils both in the absence and presence of low doses of GM-CSF. In contrast, IL-5 plus GM-CSF showed highly enhanced production of eosinophils, and eosinophil production maximized at a concentration between 10 and 20 U/ml and decreased at higher concentrations. When IL-3 and GM-CSF were kept constant at concentrations of 20 U/ml and 10 U/ml, respectively, the number of eosinophils increased linearly in IL-5-dependent manner in a range from 0.3 to 30 U/ml. These results suggest that IL-3 and GM-CSF act synergistically with IL-5 in in vitro eosinophil differentiation. In addition, we propose a new method for quantifying eosinophil differentiation activity of IL-5.

[A case of hypogamma-globulinemia with thymoma (Good's syndrome) follow-up for 8 years].

A 58-year-old woman, who had a past history of left upper lobectomy with thoracoplasty for pulmonary tuberculosis and resection of thyroid cancer, was diagnosed as having a mediastinal tumor by chest X-ray examination. It was found to be a malignant thymoma (spindle cell type) after resection. The level of serum gammaglobulin, which had been low before resection, progressively decreased. Afterward, she frequently suffered from airway infections which resulted in severe bronchiecatsis. She died due to respiratory failure 8 years later. In the early stage, though the percentage of pan T cells in peripheral blood lymphocyte subsets was normal, CD4 T cells decreased and CD8 T cells increased. A decrease in helper T cells and an increase in cytotoxic T cells were especially marked. In the late stage, all T cells subsets decreased. In particular, naive T (CD45RA* CD3+ T) cells decreased markedly. However, the percentage of B cells remained normal and that of NK cells was elevated. From the findings of lymphocyte subsets and lymphocyte reactivity to PHA stimulation, it is suggested that T cell dysfunction caused hypogrammaglobulinemia in this case.

Mechanisms of eosinophilia in Toxocara canis infected mice: in vitro production of interleukin 5 by lung cells of both normal and congenitally athymic nude mice.

Mechanisms of eosinophilia were compared between in vitro bone marrow cell cultures of congenitally athymic (nu/nu) mice and their heterozygous littermates (nu/+). Cultures of 5 x 10(4) bone marrow cells using interleukin 3 (IL-3), IL-5 and granulocyte-macrophage colony-stimulating factor showed that nu/nu and nu/+ mice mimicked each other in eosinophil production both before and after infection with Toxocara canis. Eosinophil differentiating activity (EDA) was detected in media conditioned by spleen cells and lungs of T. canis infected nu/+ mice, although nu/nu mice showed EDA only in lung-conditioned medium. EDA, detected both in infected nu/nu and nu/+ mice, was inhibited by an anti-IL-5 monoclonal antibody. These results indicate that IL-5 may be produced by lung cells of both nu/nu and nu/+ mice as well as by spleen cells of nu/+ mice infected with T. canis, which is the reason why nu/nu mice infected with T. canis exhibit blood eosinophilia.

[Estimations of organ dose and health risk of the mass chest X-ray examinations in children].

In Japan, mass chest X-ray examinations are mandated by law for schoolchildren and students. Recently, questions of the justification of such X-ray examination have arisen. In this study the absorbed doses of each organ, and the health detriments from the mass chest X-ray examinations to schoolchildren and students were estimated. The doses of organs were measured by the TLDs (Mg2SiO4), slab phantom, and anthropomorphic phantom. The probability of fatal cancer, and the resultant reduction in life expectancy induced by mass chest X-ray examinations were calculated by the multiplicative risk projection model of the ICRP-1990. The absorbed doses of lung, thyroid glands, esophagus, stomach, breast, and red bone marrow in first-year elementary schoolchildren were 90, 30, 90, 60, 90, and 30 muGy, respectively, and the doses in ovaries and testes were almost nil. Each organ dose of first-year students of junior high school was about 1.5 times that for elementary schoolchildren. The total radiation-induced lifetime cancer risk of schoolchildren and students was from 0.3 x 10(-5) to 0.9 x 10(-5) per person by the multiplicative risk projection model of the ICRP-1990 and a factor of 2 for the DDREF (dose and dose rate effectiveness factor). The reduction in life expectancy by radiation induced fatal cancer was from 15 x 10(-5) years to 50 x 10(-5) years per person. The results of this study suggest that subjects of mass chest X-ray examinations should be carefully selected from the viewpoint of radiation protection.

Identification of human hair stained with oxidation hair dyes by gas chromatographic-mass spectrometric analysis.

This paper describes the gas chromatographic-mass spectrometric (GCMS) analysis of oxidation hair dyes from human hair. Diamines from the dyes were directly extracted from the hair in basic solution and aminophenols were extracted after neutralization. Both extracts were derivatised with trifluoroacetic anhydride and analysed by GCMS. Five components of oxidation hair dyes namely, p-phenylenediamine, toluene-2,5-diamine, o-aminophenol, m-aminophenol and p-aminophenol were clearly identified, whilst no other compounds originating from the hair dyes were detected. The presence and relative amounts of these dye components from hair extracts may assist in the discrimination of human hair especially in cases involving forensic science.

[Screening of drugs and chemicals by wide-bore capillary gas chromatography. II. Detection of drugs and chemicals in the blood].

This paper describes the detection limit for 23 drugs and chemicals in the blood by means of a screening method that uses a gas chromatographic system equipped with a wide-bore capillary column and a nitrogen phosphorus detector. The detection limit by this method was determined as being 1 mm of peak height at the detector's range of 100 and 8 of attenuation. Using this scale, the absolute detection limit was in the range of 1 pg for malathion and sumithion to 1 ng for meprobamate. The detection limit of drugs and chemicals in the blood was 5 ng/ml for sumithion to 8 micrograms/ml for meprobamate. Therefore, this screening method is able to detect the presence of drugs even a therapeutic-level dosages, with the exception of compounds such as haloperidol, which have extremely low therapeutic dosage levels.

[Clinical evaluation of sulbactam/cefoperazone in lower respiratory tract infections].

Clinical evaluation, safety and kinetics in serum of sulbactam/cefoperazone (SBT/CPZ) in patients with lower respiratory tract infections have been studied in a multicenter trial participated by 28 institutions in Kyushu area during a period of 13 months from March 1987 to March 1988. 1. Mean peak serum levels of SBT and CPZ in 35 patients up to 4 hours after intravenous infusion of 2 g of SBT/CPZ were 38.2 +/- 17.3 micrograms/ml for SBT and 104.3 +/- 31.4 micrograms/ml for CPZ. Serum half-lives of SBT and CPZ were 0.76 hour and 1.53 hours, respectively. These results were in similar ranges to those reported elsewhere for SBT/CPZ. 2. Serum half-lives of SBT and CPZ after intravenous infusion of 2 g of SBT/CPZ were not significantly prolonged in patients with moderate liver or kidney dysfunctions. 3. Clinical efficacy rates of SBT/CPZ in 217 patients were 93.1% (81/87) for pneumonia, 93.3% (14/15) for lung abscess, 78.9% (15/19) for acute exacerbation of chronic bronchitis, 57.1% (4/7) for diffuse panbronchiolitis, 72.4% (21/29), 74.4% (32/43) and 100% (9/9) for infections concurrent to bronchiectasis, chronic respiratory disease and pulmonary emphysema, respectively. Those were 50% (1/2) for bronchitis associated with lung cancer and 66.7% (4/6) for empyema. The overall efficacy rate was 83.4% (181/217). 4. Clinical efficacy rate of SBT/CPZ for pneumonia in patients with underlying diseases such as lung cancer, pulmonary tuberculosis and pneumoconiosis, etc, was 85.3% (29/34) and was not significantly different from the efficacy rate of 98.1% (52/53) in patients without these underlying diseases. 5. Of 30 patients who failed to respond of previous antibiotic treatments, 21 were effectively treated by SBT/CPZ. 6. Bacteriological eradication rates against Pseudomonas aeruginosa, Haemophilus influenzae and Streptococcus pneumoniae were 42.9% (9/21), 87.5% (14/16) and 100% (5/5), respectively. The overall eradication rate in all cases including polymicrobial infections was 72.8% (67/92). 7. The high levels of peak serum concentration of CPZ, and the difference between serum levels of SBT and of CPZ seemed to contribute to the high clinical efficacy. 8. Adverse reactions occurred in 2.8% (6/217) of the patients, and consisted primarily of rash and diarrhea. Laboratory abnormalities were observed in 8 patients during the study. These were elevations of S-GOT and S-GPT, and eosinophilia. 9. SBT/CPZ is a very useful drug in the treatment of lower respiratory tract infections as it has become available just in time when increase in resistant organisms to beta-lactams is notable.

A simple, rapid and simultaneous analysis of complex volatile hydrocarbon mixtures in blood using gas chromatography/mass spectrometry with a wide-bore capillary column.

A screening method for detecting volatile hydrocarbons in blood has been developed using gas chromatography/mass spectrometry with a wide-bore capillary column and a headspace method. Toluene-d8 and indan were used as the internal standards for quantitative analysis. Hydrocarbons with retention indices from 600 to 1200 were simultaneously and quantitatively detected in relatively low concentrations (0.01 microgram/ml) in reconstructed ion chromatography. This method could prove useful in forensic cases in which urgent examination of complex hydrocarbon mixtures, e.g. petroleum components, is required.