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The substances associated with PM2.5-induced inflammatory response were investigated using an elimination method. PM2.5 were heated at temperatures of 120, 250, and 360°C. The results demonstrated microbial substances such as LPS and b-glucan, and chemicals including BaP, 1,2-NQ, and 9,10-PQ were reduced drastically in PM2.5 heated at 120°C. On the other hand, DBA, 7,12-BAQ, and BaP-1,6-Q were not noticeably reduced. Most of these substances had disappeared in PM2.5 heated at 250°C and 360°C. Metals (eg, Fe, Cu, Cr, Ni) in PM2.5 exhibited a slight thermo-dependent increase. RAW264.7 macrophages with or without NAC were exposed to unheated PM2.5, oxidative stress-related and unrelated inflammatory responses were induced. PM2.5-induced lung inflammation in mice is caused mainly by thermo-sensitive substances (LPS, b-glucan, BaP, 1,2-NQ, 9,10-PQ, etc.). Also, a slight involvement of thermo-resistant substances (DBA, 7,12-BAQ, BaP-1,6-Q, etc.) and transition metals was observed. The thermal decomposition method could assist to evaluate the PM2.5-induded lung inflammation.

Several studies show that maternal conventional cigarette smoking during pregnancy has been associated with reduced sperm concentration in sons. The development of heat-not-burn (HnB) tobacco has gained a growing following. However, the effects of prenatal HnB tobacco smoking on male offspring are as yet unknown. Pregnant CD-1 mice were exposed to I-Quit-Ordinary-Smoking (IQOS) (HnB tobacco) aerosol from heat sticks, mainstream smoke from 3R4F (conventional cigarettes) or clean air, using a whole-body exposure system. Adult male offspring mice were divided into six groups: control (5- and 15-weeks-old offspring), IQOS (5 and 15-weeks-old) and 3R4F (5 and 15-weeks-old). Spermatogenesis, sperm characteristics, serum testosterone, and seminiferous tubule morphology were evaluated. Prenatal IQOS exposure increased abnormal seminiferous tubule morphology and decreased sperm production at 5 weeks, but 3R4F exposure did not. Prenatal exposure to IQOS aerosol delays sexual maturation of male offspring or adversely affects the male testicular function of the offspring more than smoke from a combustion cigarette.

Particulate matter 2.5 (

Particulate matter with aerodynamic diameter ≤2.5 μm (PM2.5 ) is generally composed of carbon nuclei associated with various organic carbons, metals, ions and biological materials. Among these components, polyaromatic hydrocarbons (PAHs) such as benzo(a)pyrene (BaP) and quinones have detrimental effects on airway epithelial cells and immunodisrupting effects, which leads to the exacerbation of respiratory allergies. The effects of PAHs and the carbon nuclei, separately as well as in combination, remain to be established. We investigated the effects of BaP, 9,10-phenanthroquinone (9,10-PQ), and 1,2-napthoquinone (1,2-NQ) and their combined effects with heated diesel exhaust particle (H-DEP) as carbon nuclei of typical PM2.5 . We exposed human airway epithelial cells (BEAS-2B), murine bone marrow-derived antigen-presenting cells (APCs), and murine splenocytes to BaP, 9,10-PQ, or 1,2-NQ in the presence and absence of H-DEP. Several important inflammatory cytokines and cell surface molecules were measured. PAHs alone did not have apparent cytotoxic effects on BEAS-2B, whereas combined exposure with H-DEP induced noticeable detrimental effects which mainly reflected the action of H-DEP itself. BaP increased CD86 expression as an APC surface molecule regardless of the presence or absence of H-DEP. None of the BaP, 9,10-PQ, or 1,2-NQ exposure alone or their combined exposure with H-DEP resulted in any significant activation of splenocytes. These results suggest that PAHs and carbon nuclei show additive effects, and that BaP with the carbon nuclei may contribute to exacerbations of allergic respiratory diseases including asthma by PM2.5 , especially via antigen-presenting cell activation.

Epidemiologic studies have revealed that Asian sand dust particles (ASDs) can affect respiratory and immune health represented by asthma. Factors responsible for the exacerbation of asthma remain unclear. The fungus Bjerkandera adusta (B.ad) and polycyclic aromatic hydrocarbons such as benzo[a]pyrene (BaP) have been identified in ASDs collected from the atmosphere when an ASD event occurred. We investigated the effects of B.ad and BaP related to ASDs on respiratory and immune systems. Bone marrow-derived antigen-presenting cells (APCs) and splenocytes from atopic prone NC/Nga mice and human airway epithelial cells were exposed to the B.ad or to BaP in the presence and absence of heated-ASDs (H-ASDs). B.ad and BaP in both the presence and absence of H-ASDs increased the expression of cell surface molecules on APCs. H-ASDs alone slightly activated APCs. The expressions induced by B.ad were higher than those induced by BaP in the presence and absence of H-ASDs. There were no remarkable effects on the activation of splenocytes or the proinflammatory responses in airway epithelial cells. These results suggest that B.ad rather than BaP contributes to the exacerbation of asthma regardless of the presence or absence of sand particles, particularly by the activation of the immune system via APCs. Copyright © 2016 John Wiley & Sons, Ltd.

Genetic and epigenetic status, including mutations of isocitrate dehydrogenase (IDH) and TP53 and methylation of O(6) -methylguanine-DNA methyltransferase (MGMT), are associated with the development of various types of glioma and are useful for prognostication. Here, using routinely available histology sections from 312 patients with diffuse gliomas, we performed immunohistochemistry using antibodies specific for IDH1 mutation, MGMT methylation status, and aberrant p53 expression to evaluate the possible prognostic significance of these features. With regard to overall survival (OS), univariate analysis indicated that an IDH1-positive profile in patients with glioblastoma (GBM), anaplastic astrocytoma (AA), anaplastic oligoastrocytoma and oligodendroglioma, or a MGMT-negative profile in patients with GBM and AA were significantly associated with a favorable outcome. Multivariate analysis revealed that both profiles were independent factors influencing prognosis. The OS of patients with IDH1-positive/MGMT-negative profiles was significantly longer than that of patients with negative/negative and negative/positive profiles. A p53 profile was not an independent prognostic factor. However, for GBM/AA patients with IDH1-negative/MGMT-negative profiles, p53 overexpression was significantly associated with an unfavorable outcome. Thus, the immunohistochemical profiles of IDH1 and MGMT are of considerable significance in gliomas, and a combination of IDH1, MGMT and p53 profiles may be useful for prognostication of GBM/AA.

Background: Recently, a cluster of patients with an intractable allergic fungal cough who were characterized by sensitization to Bjerkandera adusta was reported. In the present study, the role of Toll-like receptors and myeloid differentiation factor 88 (MyD88) in B. adusta-induced lung inflammation was investigated. Methods: Wild-type (WT), TLR2-/-,TLR4-/-, and MyD88-/- BALB/c mice were intratracheally challenged with B. adusta 4 times at 2-week intervals. Lung pathology, bronchoalveolar lavage fluid (BALF) cytological profiles, and inflammatory mediators in BALF were investigated. Bone marrow-derived macrophages (BMDM) from TLR2-/-,TLR4-/-, TLR2/4-/-, TLR7/9-/-,MyD88-/-, and WT C57BL/6J mice were stimulated with B. adusta for 12 h, and inflammatory mediators in the culture medium were measured. Results:B. adusta caused lung inflammation along with Th2 cytokine [interleukin (IL)-5 and IL-13] and eosinophil-related chemokine [eotaxin and monocyte chemotactic protein (MCP-3)] production, an increase in eosinophils in BALF, and eosinophil infiltration in the airways in WT and TLR4-/- mice. However, Th2 and eosinophil-related responses in TLR2-/- and MyD88-/- mice were low or undetectable. The induction of neutrophils and IL-6, IL-12, IL-17A, and MCP-1 in the BALF of MyD88-/- mice was attenuated compared to that in WT mice. The induction of IL-6, TNF-α, MCP-1, and macrophage inflammatory protein-1α was reduced or undetectable in B. adusta-stimulated BMDM from TLR7/9-/- and MyD88-/- mice compared to WT mice. Conclusions: These results suggest that TLR2 and the adapter protein MyD88 may play an important role in the induction of eosinophils by B. adusta. However, TLR7/9-MyD88 might be important in the induction of neutrophils and the relevant inflammatory mediators, especially IL-17A.

Nevertheless its mechanism has not been well explained yet, PM2.5 is recognized to exacerbate asthma. In the present study, the roles of toll-like receptor (TLR) 2, TLR4 and MyD88, in exacerbation of allergen-induced lung eosinophilia caused by urban PM2.5 was investigated. TLR2-, TLR4-, MyD88-deficient and WT BALB/c mice were intratracheally challenged with PM2.5 +/− ovalbumin (OVA) four times at 2-week intervals. PM2.5 increased neutrophil numbers and KC in bronchoalveolar lavage fluid and caused slight peribronchiolar inflammation in WT mice. However, these changes were attenuated, but not completely suppressed in gene-deficient mice, especially in MyD88−/− mice. In WT mice, PM2.5 + OVA exacerbated OVA-related lung eosinophilia. This exacerbation includes increase of IL-5, IL-13, eotaxin and MCP-3; infiltration of eosinophils into the airway submucosa; proliferation of goblet cells in the airway epithelium; and the production of antigen-specific IgE and IgG1 in serum. All these effects were stronger in TLR2−/− mice than in TLR4−/− mice. In MyD88−/− mice, this pro-inflammatory mediator-inducing ability was considerably weak and lung pathology was negligible. These results suggest that urban PM2.5 may exacerbate allergic inflammation in the murine lung via a TLR2/TLR4/MyD88-signaling pathway. PM2.5-bound trace microbial elements, such as lipopolysaccharide may be a strong candidate for exacerbation of murine lung eosinophilia.

Bjerkandera adusta (B. adusta) causes fungus-associated chronic cough. However, the inflammatory response is not yet fully understood. Recently, B. adusta was identified in Asian sand dust (ASD) aerosol. This study investigated the enhancing effects of ASD on B. adusta-induced lung inflammation. B. adusta was inactivated by formalin. ASD was heated to remove toxic organic substances. ICR mice were intratracheally instilled with saline, B. adusta 0.2 µg, or B. adusta 0.8 µg with or without heated ASD 0.1 mg (H-ASD), four times at 2-week intervals. Two in vitro experiments were conducted to investigate any enhancing effects using bone marrow-derived macrophages (BMDM) from Toll-like receptor (TLR) knockout mice and ICR mice. Co-exposure to H-ASD and B. adusta, especially at high doses, caused eosinophil infiltration, proliferation of goblet cells in the airway, and fibrous thickening of the subepithelial layer, and remarkable increases in expression of Th2 cytokines and eosinophil-related cytokine and chemokine expression in bronchoalveolar lavage fluid. In the in vitro study using BMDM from wild-type, TLR2-/-, and TLR4-/- mice, the TLR-signaling pathway for cytokine production caused by B. adusta was predominantly TLR2 rather than TLR4. H-ASD increased the expression of NF-κB and cytokine production by B. adusta in BMDM from ICR mice. The results suggest that co-exposure to H-ASD and B. adusta caused aggravated lung eosinophilia via remarkable increases of pro-inflammatory mediators. The aggravation of inflammation may be related, at least in part, to the activation of the TLR2-NF-κB signaling pathway in antigen presenting cells by H-ASD.

The quality and quantity of toxic materials adsorbed onto Asian sand dust (ASD) are different based on dust source regions and passage routes. The aggravating effects of two ASDs (ASD1 and ASD2) transported from the source regions of Inner Mongolia and northeast China on lung eosinophilia were compared to clarify the role of toxic materials in ASD. The ASDs contained different amounts of lipopolysaccharides (LPS) and β-glucan (ASD1ASD2) and SiO2 (ASD1ASD2). CD-1 mice were instilled intratracheally with ASD1, ASD2 and/or ovalbumin (OVA) four times at 2-week intervals. ASD1 and ASD2 enhanced eosinophil recruitment induced by OVA in the submucosa of the airway, with goblet cell proliferation in the bronchial epithelium. ASD1 and ASD2 synergistically increased OVA-induced eosinophil-relevant cytokines interleukin-5 (IL-5), IL-13 (ASD1ASD2) and chemokine eotaxin (ASD1ASD2) in bronchoalveolar lavage fluid. ASD2 aggravating effects on lung eosinophilia were greater than ASD1. The role of LPS and β-glucan in ASD2 on the production of pro-inflammatory mediators was assessed using in vitro bone marrow-derived macrophages (BMDMs) from wild type, Toll-like receptor 2-deficient (TLR2-/-), TLR4-/-, and MyD88-/- mice (on Balb/c background). ASD2-stimulated TLR2-/- BMDMs enhanced IL-6, IL-12, TNF-α, MCP-1 and MIP-1α secretion compared with ASD2-stimulated TLR4-/- BMDMs. Protein expression from ASD2-stimulated MyD88-/- BMDM were very low or undetectable. The in vitro results indicate that lung eosinophilia caused by ASD is TLR4 dependent. Therefore, the aggravation of OVA-related lung eosinophilia by ASD may be dependent on toxic substances derived from microbes, such as LPS, rather than SiO2.

Recent epidemiological studies have suggested a positive link between atopy morbidity and exposure to phthalate esters, which are environmental chemicals mainly involved in house dust. Nevertheless, experimental studies applying several allergic in vivo models (in addition to epidemiological studies) are needed to prove the precise correlation between phthalates and facilitation of the allergic response/pathophysiology. Among the phthalate esters, di-(2-ethylhexyl) phthalate (DEHP) has been widely used in flexible polyvinyl chloride products, including vinyl flooring and wall covering, and has been widely suggested to have immunomodulating potential. In the present study, we examined the effects of airway exposure to DEHP on allergen (ovalbumin: OVA)-induced rhinitis in mice. The repeated administration of OVA via an intranasal route induced nasal inflammation characterized by the infiltration of granulocytes (neutrophils and eosinophils) into the nasal cavity. In this experimental setting, DEHP did not exaggerate OVA-related inflammatory pathology. However, local (nasal) IL-13 levels were significantly higher in mice treated with allergen plus DEHP than with allergen alone. Taken together, phthalate esters including DEHP have the potential to exacerbate the allergic milieu in the nasal system, as well as dermal and respiratory systems.

Epidemiologic studies have reported that Asian sand dust (ASD) particles can affect respiratory health; however, the mechanisms remain unclear. We investigated the effects of ASD on airway epithelial cells and immune cells, and their contributing factors to the effects. Human airway epithelial cells were exposed to ASD collected on 1-3 May (ASD1) and on 12-14 May (ASD2) 2011 in Japan and heat-treated ASD1 for excluding heat-sensitive substances (H-ASD) at a concentration of 0, 3, 30 or 90 µg ml(-1) for 4 or 24 h. Furthermore, bone marrow-derived dendritic cells (BMDC) from atopic prone mice were differentiated by culture with granulocyte-macrophage colony-stimulating factor (GM-CSF) then these BMDC were exposed to the ASD for 24 h. Also splenocytes as mixture of immune cells were exposed to the ASD for 72 h. All ASD dose dependently reduced viability of airway epithelial cells. Non-heated ASD showed a dose-dependent increase in the protein release of interleukin (IL)-6 and IL-8. The raises induced by ASD1 were higher than those by ASD2. ASD1 and ASD2 also elevated ICAM-1 at the levels of mRNA, cell surface protein and soluble protein in culture medium. In contrast, H-ASD did not change most of these biomarkers. Non-heated ASD showed enhancement in the protein expression of DEC205 on BMDC and in the proliferation of splenocytes, whereas H-ASD did not. These results suggest that ASD affect airway epithelial cells and immune cells such as BMDC and splenocytes. Moreover, the difference in ASD events and components adhered to ASD can contribute to the health effects.

Urban particulate matter (PM) is associated with an increase in asthma. PM2.5 (

In recent experimental studies, we reported the aggravating effects of Asian sand dust (ASD) on male reproductive function in mice. However, the effects of fetal ASD exposure on male reproductive function have not been investigated. The present study investigated the effects of fetal ASD exposure on reproductive function in male offspring. Using pregnant CD-1 mice, ASD was administered intratracheally on days 7 and 14 of gestation, and the reproductive function of male offspring was determined at 5, 10, and 15 weeks after birth. The secondary sex ratio was significantly lower in the fetal ASD-exposed mice than in the controls. Histologic examination showed partial vacuolation of seminiferous tubules in immature mice. Moreover, daily sperm production (DSP) was significantly less in the fetal ASD-exposed mice than in the controls. DSP in the fetal ASD-exposed mice was approximately 10% less than the controls at both 5 and 10 weeks. However, both the histologic changes and the DSP decrease were reversed as the mice matured. These findings suggest that ASD exposure affects both the fetal development and the reproductive function of male offspring. In the future, it will be necessary to clarify the onset mechanisms of ASD-induced male fetus death and male reproductive disorders.

Asian sand dust (ASD) event may result in a significant influence on an asthmatic patient. However, for obvious reasons, there is no experimental study in which asthmatic patients are exposed to ASD. This study was undertaken to clarify the effects of ASD on lung eosinophiliain mice immunized beforehand by ovalbumin (OVA). CD-1 mice were instilled intratracheally with OVA four times at 2-week intervals. Simultaneous intratracheal administration of OVA and ASD (OVA + ASD sim) at the last OVA treatment or intratracheal administration with ASD 1 day before (OVA + ASD pre) /after (OVA + ASD post) the last OVA treatment was performed to investigate the effects of OVA and ASD exposure timing. The three kinds of treatment (OVA + ASD pre; OVA + ASD sim; OVA + ASD post) aggravated allergic lung inflammation and proliferation of goblet cells in the airway epithelium in mice, as evidenced by the cellular profile of bronchoalveolar lavage fluid (BALF) and pathological examination. As an overall trend, these changes were paralleled with the expression of Th2-associated effecter molecules and eosinophil relevant cytokine chimokines in BALF as well as the production of OVA-specific IgG1 compared with OVA treatment alone. OVA + ASD sim aggravated lung eosinophilia remarkably compared with the other treatments. The order of the potency of the aggravation was OVA+ASD preOVA+ASD postOVA+ASD sim. These results indicate that ASD has a potent effect in activating lung eosinopilia in mice immunized beforehand by OVA. The simultaneous exposure of asthmatic patients to ASD and its antigen may have serious consequences for such individuals.

Urban particulate matter (UPM) has been shown to have an aggravating effect on Th2-associated immune systems in adult mice. However, the effects of fetal exposure to UPM on immune response in offspring have not been elucidated. In the present study, we administered UPM (200 µg/animal) by intratracheal injection to pregnant dams on days 7 and 14 of gestation. Subsequently, 9- and 24-week-old male offspring were intratracheally injected with ovalbumin (OVA) (four times at 2-week intervals) to create a mouse model of bronchial asthma. We then evaluated the progression of allergic manifestations in the offspring through histological findings, the number of inflammatory cells in bronchoalveolar lavage fluid (BALF), and protein concentration of cytokines and chemokines in BALF 5, 10, 15, and 30 weeks after birth. Histological examination showed that fetal exposure to UPM alone caused slight eosinophil and lymphocyte infiltration in the submucosa of the airway and bronchial epithelium and significant increases in the number of macrophages. Moreover, postnatal intratracheal administration of OVA to offspring exposed to UPM in utero caused significant increases in the numbers of macrophages, eosinophils, and lymphocytes and in the concentrations of their relevant cytokines and chemokines, showing that fetal exposure to UPM aggravated the chemically sensitized immune system of male offspring.

Objective To investigate the effects of fetal nanoparticle exposure on reproductive function in male mice offspring. Design Animal study. Setting Academic research laboratory. Animal(s) Forty pregnant ICR mice and 120 male offspring. Intervention(s) Two hundred μg of 14-nm carbon nanoparticles was administered intratracheally on days 7 and 14 of gestation, and reproductive function of male offspring was determined at ages 5, 10, and 15 weeks after birth. Main Outcome Measure(s) Maternal and fetal growth, histologic changes in the testes, and daily sperm production (DSP). Result(s) Histologic examination showed partial vacuolation of seminiferous tubules. and cellular adhesion of seminiferous epithelia was reduced at all three ages. In addition, DSP was significantly decreased in fetal carbon nanoparticle–exposed mice. The DSP in the fetal carbon nanoparticle–exposed mice decreased by 47% at the age of 5 weeks, by 34% at the age of 10 weeks, and by 32% at the age of 15 weeks. On the other hand, nanoparticle administration had no marked effect on body weight, testicle weight, epididymis weight, or serum testosterone concentration. Conclusion(s) These findings suggest that fetal nanoparticle exposure affects the reproductive function of male offspring. In the future, it would be necessary to clarify the onset mechanisms of nanoparticle-induced male reproductive disorders.

我々はこの20年間における1施設での, 視床ならびに被殻部の悪性神経膠腫症例をステージ分類し, この部位における温熱治療症例の臨床的特徴について報告する. 対象は, 新潟大学において治療を受けた基底核部 (視床を含む) のグレード3および4の悪性グリオーマ43例である. 病期分類は, 視床並びに被殻部の腫瘍の大きさを基に行った. すなわち, ステージ1 (2cm未満), ステージ2 (2―4cm), ステージ3 (4cm以上) とし, 腫瘍が被殻や視床を超えて他部位へ進展しているものをステージ4, 脳室への進展例 (播種例) をステージ5として分類した. 当科における温熱治療は, 腫瘍内に電極を局所麻酔下に生検する際に同時に埋め込んでくるもので, 通常の外照射60Gyとともに週2回計3ないし4回の加温を行うものである. 化学療法はACNUまたはMCNUを温熱放射線治療中に静注もしくは動注しているが, 高齢者においては併用していない. その結果, ステージ4症例は全症例の30％を占め, ステージ5症例も含めると40％にもなった. 摘出術施行例は43例中14例で, 残りの29例は生検術症例である. そのうちの14例は生検術とともに温熱治療のための針電極を留置し, 放射線治療とともに温熱治療を行った. 平均生存期間は11ヶ月であった (視床部では17ヶ月, 被殻部で11ヶ月). 視床部の悪性グリオーマは被殻部の症例に比し, 若年である傾向が認められ, 予後も若干良好であった. 温熱症例の平均生存期間は22か月で, 非温熱治療, 生検術群の9ヶ月に比べ, 生存期間延長の傾向が認められた. 基底核部悪性グリオーマは深部であるため, 摘出術が可能であることが少なく予後不良であったが, 温熱治療により有効例も認められるようになっている.

The effect of sample collection site on semen parameters in ten men aged between 22 and 24 years was investigated. Sperm was collected at two sites: in a university hospital restroom for general use and in a one-person hospital room. Samples were collected from the same individual twice, with an interval of two weeks between collections. Semen parameters for the two sites were compared. Samples were collected after a minimum of three days and not longer than seven days of sexual abstinence. Sperm concentration did not differ significantly between the university hospital restroom location (86.8 +/- 25.4 x 10(6)/ml; mean +/- standard deviation) and the private hospital room (97.1 +/- 72.0 x 10(6)/ml). There was no difference in the total motile sperm count or daily sperm production between the collection sites. These results suggest that the collection site has little effect on semen parameters.

The biological and chemical natures of materials adsorbed onto fine particulate matter (PM2.5) vary by origin and passage routes. The exacerbating effects of the two samples-urban PM2.5 (U-PM2.5) collected during the hazy weather in a Chinese city and fine particles (ASD-PM2.5) collected during Asian sand dust (ASD) storm event days in Japan-on murine lung eosinophilia were compared to clarify the role of toxic materials in PM2.5. The amounts of β-glucan and mineral components were higher in ASD-PM2.5 than in U-PM2.5. On the other hand, organic chemicals, including polycyclic aromatic hydrocarbons (PAHs), were higher in U-PM2.5 than in ASD-PM2.5. When BALB/c mice were intratracheally instilled with U-PM2.5 and ASD-PM2.5 (total 0.4 mg/mouse) with or without ovalbumin (OVA), various biological effects were observed, including enhancement of eosinophil recruitment induced by OVA in the submucosa of the airway, goblet cell proliferation in the bronchial epithelium, synergic increase of OVA-induced eosinophil-relevant cytokines and a chemokine in bronchoalveolar lavage fluid, and increase of serum OVA-specific IgG1 and IgE. Data demonstrate that U-PM2.5 and ASD-PM2.5 induced allergic inflammatory changes and caused lung pathology. U-PM2.5 and ASD-PM2.5 increased F4/80(+) CD11b(+) cells, indicating that an influx of inflammatory and exudative macrophages in lung tissue had occurred. The ratio of CD206 positive F4/80(+) CD11b(+) cells (M2 macrophages) in lung tissue was higher in the OVA+ASD-PM2.5 treated mice than in the OVA+U-PM2.5 treated mice. These results suggest that the lung eosinophilia exacerbated by both PM2.5 is due to activation of a Th2-associated immune response along with induced M2 macrophages and the exacerbating effect is greater in microbial element (β-glucan)-rich ASD-PM2.5 than in organic chemical-rich U-PM2.5.

Asian sand dust (ASD) is known to exacerbate asthma, although its mechanism is not yet well understood. In this study, when the effects on inflammatory response by LPS present in ASD was investigated by measuring the gene expression of cytokines and chemokines in RAW264.7 cells treated with ASD and/or polymyxin B (PMB), the ASD effects were attenuated by PMB, but not completely. When an in vitro study was performed using bone marrow-derived macrophages (BMDMs) from WT, TLR2(-/-), TLR4(-/-), and MyD88(-/-) BALB/c mice and BMDMs from WT, TLR2(-/-), TLR4(-/-), TLR2/4(-/-), TLR7/9(-/-), and MyD88(-/-) C57BL/6J mice, cytokine (IL-6, IL-12) production in BMDMs was higher in ASD-stimulated TLR2(-/-) cells than in TLR4(-/-) cells, whereas it was lower or undetectable in TLR2/4(-/-) and MyD88(-/-) cells. These results suggest that ASD causes cytokine production predominantly in a TLR4/MyD88-dependent pathway. When WT and TLRs 2(-/-), 4(-/-), and MyD88(-/-) BALB/c mice were intratracheally challenged with OVA and/or ASD, ASD caused exacerbation of lung eosinophilia along with Th2 cytokine and eosinophil-relevant chemokine production. Serum OVA-specific IgE and IgG1 similar to WT was observed in TLRs 2(-/-), 4(-/-) mice, but not in MyD88(-/-) mice. The Th2 responses in TLR2(-/-) mice were attenuated remarkably by PMB. These results indicate that ASD exacerbates lung eosinophilia in a MyD88-dependent pathway. TLRs 2 and 4 signaling may be important in the increase in lung eosinophilia. Also, the TLR4 ligand LPS and TLR2 ligand like β-glucan may be strong candidates for exacerbation of lung eosinophilia.

Background: Bisphenol A (BPA) is useful in many manufacturing processes and is also found in commonly used consumer products. Previous experimental studies have reported that perinatal exposure to BPA promotes the development of allergic lung inflammation in childhood and even into adulthood. In this study, the effects of BPA on allergic lung inflammation in adults were investigated in murine lungs. Methods: CD-1 mice were orally administrated with 1 mg of BPA/mouse four times at one-week intervals with or without ovalbumin (OVA). The pathologic changes in the airways, cytological alterations in bronchoalveolar lavage fluid (BALF), levels of inflammatory cytokines/chemokines in BALF, and OVA-specific IgE and IgG1 antibodies in serum were measured in the treated CD-1 mice. In vitro study using RAW264.7 cells, which are macrophage-like cells derived from BALB/c male mice, was conducted. The gene expression of cytokines and chemokines were measured. Results: BPA enhanced eosinophil recruitment induced by OVA in the alveoli and in the submucosa of the airway, which has a goblet cell proliferation in the bronchial epithelium. BPA increased Th2 cytokines-interleukin-13 (IL-13), eosinophil-relevant cytokines and chemokines, such as IL-5, and CCL2 induced by OVA, in BALF. BPA induced adjuvant effects on OVA-specific IgG1 production. In the in vitro study using RAW264.7 cells, BPA increased the mRNA expression of IL-1β, IL-6, CCL2 and CCL3 compared with the control and OVA groups. Conclusions: These results suggest that (1) the exposure of BPA could synergize with an OVA challenge to aggravate the severity of lung eosinophilia in adult mice, possibly by promoting a Th2-biased immune response and (2) the activation of macrophages and inflammatory cytokines released from these cells by BPA could be participating in this phenomenon.

Recently, a cluster of patients with an intractable allergic fungal cough who were characterized by sensitization to Bjerkandera adusta was reported. In the present study, the role of Toll-like receptors and myeloid differentiation factor 88 (MyD88) in B. adusta-induced lung inflammation was investigated.Wild-type (WT), TLR2-/-,TLR4-/-, and MyD88-/- BALB/c mice were intratracheally challenged with B. adusta 4 times at 2-week intervals. Lung pathology, bronchoalveolar lavage fluid (BALF) cytological profiles, and inflammatory mediators in BALF were investigated. Bone marrow-derived macrophages (BMDM) from TLR2-/-,TLR4-/-, TLR2/4-/-, TLR7/9-/-,MyD88-/-, and WT C57BL/6J mice were stimulated with B. adusta for 12 h, and inflammatory mediators in the culture medium were measured.B. adusta caused lung inflammation along with Th2 cytokine [interleukin (IL)-5 and IL-13] and eosinophil-related chemokine [eotaxin and monocyte chemotactic protein (MCP-3)] production, an increase in eosinophils in BALF, and eosinophil infiltration in the airways in WT and TLR4-/- mice. However, Th2 and eosinophil-related responses in TLR2-/- and MyD88-/- mice were low or undetectable. The induction of neutrophils and IL-6, IL-12, IL-17A, and MCP-1 in the BALF of MyD88-/- mice was attenuated compared to that in WT mice. The induction of IL-6, TNF-α, MCP-1, and macrophage inflammatory protein-1α was reduced or undetectable in B. adusta-stimulated BMDM from TLR7/9-/- and MyD88-/- mice compared to WT mice.These results suggest that TLR2 and the adapter protein MyD88 may play an important role in the induction of eosinophils by B. adusta. However, TLR7/9-MyD88 might be important in the induction of neutrophils and the relevant inflammatory mediators, especially IL-17A.

Background Meningeal carcinomatosis (MC) is increasing, and these patients have a poor prognosis. We analyzed the effects of intrathecal (IT) chemotherapy for these patients. Methods Patients received both methotrexate (MTX) (15 mg/m 2 ) and prednisolone (10 mg/m 2 ) 6 times in 2 weeks by Ommaya reservoir, along with cytosine arabinoside (10 mg/m 2 ) for 4 doses of MTX. A cycle consisted of a 2-week period during which patients received these drugs and then 2 weeks off. Treatments were repeated 3 to 6 cycles depending on the clinical status. Cerebrospinal fluid (CSF) samples were also analyzed for cytology and a few markers. Results Of the 58 patients treated the most common tumor was lymphoma (30 patients), followed by lung and breast. Elevated soluble IL-2 receptor levels were observed in 23 of 30 patients with lymphomatous meningitis. Median survival of MC patients with malignant lymphoma, lung cancer, and breast cancer was 32.8 ± 9.8, 13.0 ± 4.1, and 18.4 ± 7.4 months, respectively. Thus, the patients with lymphoma responded best, both by clearing the CSF and clinically. Conclusions Our treatment regimen can improve the neurologic status of patients with MC. In particular, early IT chemotherapy can be effective for patients with lymphoma.

We report on the discovery of a new variable star, MisV1147, by the MISAO Project and its unusual photometric variations. The light curve of the object is characterized by two distinct states, that is, a relatively calm, bright state and an active state during which repetitive fadings with large amplitudes of ∼2mag were observed. The fading events were associated with optical reddening. The object is identified with a catalogued Hα emission star, which has been confirmed by our spectroscopic observations. The Hα emission line is narrow and symmetric both in the bright and faint phases. The object has infrared colors analogous to T Tau stars or Herbig Ae/Be stars. In conjunction with the detection of a NaD absorption line and its position close to Cep OB1, we conclude that MisV1147 is a Herbig Be star with its spectral type of B2. Its optical-infrared spectral energy distribution is typical for Herbig Ae/Be stars, which supports this classification. The overall properties of the light curve are reminiscent of a UX Ori sub-group, showing Algol-type minima. The most noteworthy feature of MisV1147 is that it has a spectral type B2, whereas known UX Ori stars have spectral types later than B8. The object, furthermore, has one of the largest amplitudes of variations among known Herbig Ae/Be stars. Hence the object breaks the very strong correlation between the variation amplitude and the spectral type. This implies that dust clumps in circumstellar matter can significantly obscure a central object even in the early-type system.

Several reports have suggested that semen quality in normal men is declining. Given the lack of consensus about the effects of diesel exhaust (DE) on the male reproductive system, we conducted various experiments. We examined the effect of the exposure of mature male mice to DE for 6 months on the male reproductive system. Daily sperm production per gram from the testes dose-dependently decreased with exposure to DE. Next, we investigated the effect of exposure of pregnant mice to DE on male gonad development at the level of mRNA expression. Expression of mRNAs for steroidogenic factor (Ad4BP/SF-1) and Mullerian inhibiting substance (MIS) decreased significantly in male fetuses when maternal mice were exposed to DE for 8 hr per day between days 2 and 13 post coitum. In addition, DE exposure during the fetal period may have some influence on the male reproductive function in newborn mice. In utero DE exposure caused testosterone levels to increase in newborn male mice at age 4 weeks. These findings indicate that exposure to DE may influence the male reproductive system. Further studies are necessary to elucidate the mechanism of the effects of DE on the male reproductive system and to define which classes of compounds are responsible for the changes in the male reproductive system.

Individual variation in semen parameters was investigated in healthy young volunteers. Semen samples were collected approximately once a month over a one-year period for a total of 93 samples (5 to 10 samples per subject) from 12 volunteers in their twenties. Semen analysis was carried out according to the WHO Manual. The amount of variation in each semen variable was calculated for each subject by dividing the maximum value by the minimum value. The results showed that the semen volume varied by 1.9+/-0.8 fold (1.3 to 4.2 fold), the sperm concentration by 4.8+/-4.3 fold (1.5 to 17.2 fold), the percentage of sperm with forward progression by 2.8+/-1.4 fold (1.6 to 6.4 fold), the percentage of sperm with rapid linear progression by 3.4+/-2.6 fold (1.7 to 10.9 fold), the percentage of sperm with normal morphology by 1.9+/-0.4 fold (1.3 to 2.4 fold), and the percentage of live sperm by 1.5+/-0.4 fold (1.1 to 2.6 fold). A between-group comparison showed significant differences in all of the variables except the percentage of sperm with normal morphology. These results suggest multiple and considerable semen analyses are needed when evaluating semen parameters.

Diesel exhaust (DE) is a serious air pollution problem in big cities. Most suspended particulate matter (SPM) less than 2.5 μm in diameter consists of diesel exhaust particles (DEPs), which are reported to cause pulmonary carcinogenesis, allergic rhinitis, and bronchial asthma-like diseases. It has been recently reported that DE also affects the circulatory and reproductive systems. Yoshida et al. reported that mRNA expression of steroidogenic factor-1 (Ad4BP/SF-1) and of Mullerian inhibitory substance (MIS), which are essential for male gonadal differentiation, decreased significantly in male fetuses when maternal mice were exposed to DE at levels of 0.1 or 3.0 mg DEP/m3 for 8 hr per day between 2 and 13 days postcoitum (dpc). In this study, maternal mice were exposed to DE 0.1 mg DEP /m3 for 8 hr per day between 2 and 13 dpc. Expression levels of Ad4BP/SF-1 and MIS mRNA in female fetuses were not decreased. However, expression levels of bone morphogenetic protein-15, reported to be related to development of the oocyte, were significantly decreased in comparison with that in the control group. Our data suggest that female fetuses of pregnant mice exposed to DE in utero are less sensitive to the expression levels of mRNAs for Ad4BP/SF-1 and MIS compared with males and that DE may affect development of the oocyte in the female fetus.

The initial processes of nitridation of silicon thin film have been examined by means of the first-principle calculations. The nitrogen atom on the bare silicon(1 0 0) surface stays around the surface due to the dangling bond of the first-layer silicon, and the large stabilization on the bare silicon surface raises the energy barrier of the penetration of the nitrogen atom into the surface. The nitrogen atom on the H-terminated silicon surface can penetrate into the surface by crossing over the small energy barrier. In the nitridation by using nitrous oxide (N 2 O) molecule, N 2 O can be adsorbed only with the large NN bond length, and then an exothermic elimination of nitric oxide (NO) molecule from the adsorbed nitrogen atom on the bare silicon surface takes place. The electronic processes have been discussed in terms of the quantum mechanical energy densities, which give new images of microscopic electronic stresses in the interaction system.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is an endocrine disruptor that is known to have widespread effects on the reproductive system. We examined the effects of TCDD on testosterone production of primary cultured murine testicular cells. The cells, derived from the testes of ICR mice at age 65 days, were exposed to 10-2-104 pM TCDD for 3, 24, or 48 hr and treated with human chorionic gonadotropin for 6 hr to induce production of testosterone. At these concentrations of TCDD, the viability of testicular cells was not affected. No significant time- or TCDD concentration-dependent effects were observed on the secretion of testosterone. The data suggest that TCDD does not have a direct influence on testosterone production in the ICR mouse.

Genetic and epigenetic status, including mutations of isocitrate dehydrogenase (IDH) and TP53 and methylation of O(6) -methylguanine-DNA methyltransferase (MGMT), are associated with the development of various types of glioma and are useful for prognostication. Here, using routinely available histology sections from 312 patients with diffuse gliomas, we performed immunohistochemistry using antibodies specific for IDH1 mutation, MGMT methylation status, and aberrant p53 expression to evaluate the possible prognostic significance of these features. With regard to overall survival (OS), univariate analysis indicated that an IDH1-positive profile in patients with glioblastoma (GBM), anaplastic astrocytoma (AA), anaplastic oligoastrocytoma and oligodendroglioma, or a MGMT-negative profile in patients with GBM and AA were significantly associated with a favorable outcome. Multivariate analysis revealed that both profiles were independent factors influencing prognosis. The OS of patients with IDH1-positive/MGMT-negative profiles was significantly longer than that of patients with negative/negative and negative/positive profiles. A p53 profile was not an independent prognostic factor. However, for GBM/AA patients with IDH1-negative/MGMT-negative profiles, p53 overexpression was significantly associated with an unfavorable outcome. Thus, the immunohistochemical profiles of IDH1 and MGMT are of considerable significance in gliomas, and a combination of IDH1, MGMT and p53 profiles may be useful for prognostication of GBM/AA.

Background The organic chemicals present in Asian sand dust (ASD) might contribute to the aggravation of lung eosinophila. Therefore, the aggravating effects of the Tar fraction from ASD on ovalbumin (OVA)-induced lung eosinophilia were investigated. Methods The Tar fraction was extracted from ASD collected from the atmosphere in Fukuoka, Japan. ASD collected from the Gobi desert was heated at 360°C to inactivate toxic organic substances (H-ASD). ICR mice were instilled intratracheally with 12 different test samples prepared with Tar (1 μg and 5 μg), H-ASD, and OVA in a normal saline solution containing 0.02% Tween 80. The lung pathology, cytological profiles in the bronchoalveolar lavage fluid (BALF), inflammatory cytokines/chemokines in BALF and OVA-specific immunoglobulin in serum were investigated. Results Several kinds of polycyclic aromatic hydrocarbons (PAHs) were detected in the Tar sample. H-ASD + Tar 5 μg induced slight neutrophilic lung inflammation. In the presence of OVA, Tar 5 μg increased the level of eosinophils slightly and induced trace levels of Th2 cytokines IL-5 and IL-13 in BALF. Also mild to moderate goblet cell proliferation and mild infiltration of eosinophils in the submucosa of airway were observed. These pathological changes caused by H-ASD + OVA were relatively small. However, in the presence of OVA and H-ASD, Tar, at as low a level as 1 μg, induced severe eosinophil infiltration and proliferation of goblet cells in the airways and significantly increased Th2 cytokines IL-5 and IL-13 in BALF. The mixture showed an adjuvant effect on OVA-specific IgG1 production. Conclusions These results indicate that H-ASD with even low levels of Tar exacerbates OVA-induced lung eosinophilia via increases of Th2-mediated cytokines. These results suggest that ASD-bound PAHs might contribute to the aggravation of lung eosinophila.

Bjerkandera adusta (B. adusta) causes fungus-associated chronic cough. However, the inflammatory response is not yet fully understood. Recently, B. adusta was identified in Asian sand dust (ASD) aerosol. This study investigated the enhancing effects of ASD on B. adusta-induced lung inflammation. B. adusta was inactivated by formalin. ASD was heated to remove toxic organic substances. ICR mice were intratracheally instilled with saline, B. adusta 0.2 µg, or B. adusta 0.8 µg with or without heated ASD 0.1 mg (H-ASD), four times at 2-week intervals. Two in vitro experiments were conducted to investigate any enhancing effects using bone marrow-derived macrophages (BMDM) from Toll-like receptor (TLR) knockout mice and ICR mice. Co-exposure to H-ASD and B. adusta, especially at high doses, caused eosinophil infiltration, proliferation of goblet cells in the airway, and fibrous thickening of the subepithelial layer, and remarkable increases in expression of Th2 cytokines and eosinophil-related cytokine and chemokine expression in bronchoalveolar lavage fluid. In the in vitro study using BMDM from wild-type, TLR2-/-, and TLR4-/- mice, the TLR-signaling pathway for cytokine production caused by B. adusta was predominantly TLR2 rather than TLR4. H-ASD increased the expression of NF-κB and cytokine production by B. adusta in BMDM from ICR mice. The results suggest that co-exposure to H-ASD and B. adusta caused aggravated lung eosinophilia via remarkable increases of pro-inflammatory mediators. The aggravation of inflammation may be related, at least in part, to the activation of the TLR2-NF-κB signaling pathway in antigen presenting cells by H-ASD.

Background Bjerkandera adusta (B. adusta) is one of the most important etiological fungi associated with chronic cough. However, precise details of the inflammatory response to exposure are not well understood yet. B. adusta was recently identified in Asian sand dust (ASD) aerosol. Therefore, in the present study the exacerbating effects of ASD on B. adusta-induced lung inflammation and B. adusta + ASD on ovalbumin (OVA)-induced murine lung eosinophilia were investigated using experimental mice. Methods In order to prepare testing samples, B. adusta obtained from ASD aerosol was inactivated by formalin and ASD collected from the atmosphere was heated to remove toxic organic substances (H-ASD). CD-1 mice were instilled intratracheally with 12 different samples prepared with various combinations of B. adusta, H-ASD, and OVA in a normal saline solution. The lung pathology, cytological profiles in bronchoalveolar lavage fluid (BALF), and the levels of inflammatory cytokines/chemokines in BALF were investigated. Results H-ASD aggravated the lung eosinophilia induced by B. adusta alone, which also aggravated the lung eosinophilia induced by OVA. The mixture of OVA, H-ASD, and B. adusta caused serious fibrous thickening of the subepithelial layer, eosinophil infiltration, and proliferation of goblet cells in the airways along with remarkable increases of IL-13, eotaxin, IL-5, and MCP-3 in BALF. Conclusions The results of the present study demonstrated that B. adusta isolated from ASD aerosol induces allergic lung diseases. H-ASD enhanced allergic reactions caused by OVA or B. adusta. A mixture of B. adusta, H-ASD, and OVA caused the most remarkable exacerbation to the allergic airway inflammation via remarkable increases of pro-inflammatory mediators.

BackgroundA previous study has shown that the aggravation of Asian sand dust (ASD) on ovalbumin (OVA)-induced lung eosinphilia was more severe in lipopolysaccharide (LPS)-rich ASD than in SiO2-rich ASD. Therefore, the effects of different LPS contamination levels in ASD on the aggravation of OVA-induced lung eosinophilia were investigated in the present study.MethodsBefore beginning the in vivo experiment, we investigated whether the ultra-pure LPS would act only on TLR4 or not using bone marrow-derived macrophages (BMDMs) of wild-type, TLR2-/-, TLR4-/- and MyD88-/- BALB/c mice. ASD collected from the desert was heated to remove toxic organic substances (H-ASD). BALB/c mice were instilled intratracheally with 12 different testing samples prepared with LPS (1ng and 10ng), H-ASD, and OVA in a normal saline solution. The lung pathology, cytological profiles in the bronchoalveolar lavage fluid (BALF), the levels of inflammatory cytokines/chemokines in BALF and OVA-specific immunoglobulin in serum were investigated.ResultsThe LPS exhibited no response to the production of TNF-α and IL-6 in BMDMs from TLR4-/-, but did from TLR2-/-. H-ASD aggravated the LPS-induced neutrophilic lung inflammation. In the presence of OVA, LPS increased the level of eosinophils slightly and induced trace levels of Th2 cytokines IL-5 and IL-13 at the levels of 1ng and 10ng. In the presence of OVA and H-ASD, LPS induced severe eosinophil infiltration and proliferation of goblet cells in the airways as well as remarkable increases in Th2 cytokines IL-5 and IL-13 in BALF. The mixture containing LPS (1ng) showed adjuvant activity on OVA-specific IgE and IgG1 production.ConclusionsThe results suggest that H-ASD with naturally-occurring levels of LPS enhances OVA-induced lung eosinophilia via increases in Th2-mediated cytokines and antigen-specific immunoglobulin. These results indicate that LPS is a strong candidate for being a major aggravating substance in ASD.

Dendritic cells (DCs) can be the principal initiators of antigen-specific immune responses. We analyzed the in vitro-responses against brain tumor cells using DCs from the peripheral blood of patients with brain tumors. Peripheral blood mononuclear cells (PBMC) were obtained from 19 patients with malignant brain tumors: 12 metastatic brain tumors of lung adenocarcinoma, 7 high-grade astrocytomas. PBMC were cultured with 100 ng/ml of GM-CSF and 10 ng/ml of IL-4 for 5 7 days in order to produce mature DCs. The autologous tumor lysate (5 mg/ml, containing 1 x 10(6) cells) was then added to the cultured DCs. Using the DCs generated by these treatments, we assessed the changes that occurred in their immune responses against brain tumor via 51Cr-release and lymphocyte proliferation assays. We found that the matured DCs displayed the typical surface phenotype of CD3+, CD45+, CD80+ and CD86+. After the pulsation treatment with tumor lysate, DCs were found to have strong cytotoxic T lymphocyte activity, showing 42.5+12.7% killing of autologous tumor cells. We also found an enhancement of allogeneic T cell proliferation after pulsing the DC with tumor lysate. These data support the efficacy of DC-based immunotherapy for patients with malignant brain tumors.

セミミクロLCに, 内径3mmのプローブを組み込んだ400MHzのNMRを接続し, セミミクロLC/NMR装置を開発した。セミミクロカラムには内径1.5mm長さ25cmのカラムを用い, 移動相に重水と重アセトニトリルの混液を流量100-125μL/minで用いて種々の検討を行った。本装置は, 1時間あたりわずか6-7.5mLの移動相しか必要とせず, その優れたコストパフォーマンスにより, 高価な重水素化溶媒を移動相に使用できるので, 容易に移動相信号を極小化することが可能となった。また, セミミクロLCの濃縮効果による感度の向上が得られ, セミミクロLC/NMRの優れた性能が示された。本法の, p-ヒドロキシ安息香酸エチルを試料としたときの解析限界は, ON-FLOW法でμgオーダー, STOP & FLOW法でサブμgオーダーであった。また, 化粧品原料分析への応用として, 生薬エキス末中の成分分析へ適用したところ, 分取することなく含有成分をモニターできたことにより, 本システムが迅速な有効成分の探索に有用であることが実証された。

The antigen-presenting capability of syngeneic rat glial cells was investigated under glioma-harboring conditions. Microglia induced a significant proliferation of glioma-primed splenocytes, but astrocytes did not. Furthermore, astrocytes suppressed the accessory cell function of microglia. The presence of both indomethacin and anti-interleukin (IL)-10 neutralizing antibody during priming of microglia enhanced splenocyte proliferation. The glioma culture supernatants down-regulated the interferon-γ-induced expression of major histocompatibility complex class II molecules on microglia. The down-regulation was blocked by indomethacin and anti-IL-10 antibody. The results suggest that microglia but not astrocytes may function as antigen-presenting cells in glioma, and that glioma may suppress the antigen-presenting abilities of microglia.

BACKGROUND The clinical characteristics of brain metastasis from sarcoma are not well known. We examined the incidence and the characteristics of brain metastasis in patients with sarcomas. METHODS All sarcoma patients treated at our institution from 1975 to 1998 were reviewed for brain metastasis. Diagnosis of the primary tumor was confirmed histologically, and brain metastasis was confirmed using computed tomographic (CT) brain scan. RESULTS Brain metastasis was found in 27 (5.6%) of 480 patients with systemic sarcoma (7.2% soft part sarcoma, 3.5% bone sarcoma, 15.1% distant metastasis). Of these 27 sarcoma patients with brain metastases, lung metastasis occurred in 16 patients (59.3%). Out of 10 patients surgically treated, 8 patients survived more than 16 months. Median survival period after craniotomy was 25.4 months. CONCLUSIONS We recommend aggressive treatment for those patients with brain metastases whose performance scores are over 70.

BACKGROUND AND OBJECTIVES Interleukin-2 receptor alpha (IL-2Ralpha) can combine with IL-2 firmly, and soluble IL-2Ralpha (sIL-2Ralpha) is elevated in sera from patients with various types of cancers. To investigate the role of this receptor, we studied the changes of serum sIL-2Ralpha in patients with malignant brain tumors. METHODS SIL-2Ralpha was measured in 100 patients with malignant brain tumors (63 cancer metastasis, 16 malignant gliomas, 21 malignant lymphomas), and 51 patients with cancer who had no distant metastasis such as brain metastasis. RESULTS In patients with 35 metastatic brain tumors from lung cancer, the levels of sIL-2Ralpha were not significantly different from levels in normal volunteers (311 +/- 62.4 U/ml). In patients with 25 metastatic brain tumors from lung adenocarcinoma, the mean level of serum sIL-2Ralpha was 352 +/- 94.0 U/ml. These same patients showed high levels of serum sIL-2Ralpha (492 +/- 101 U/ml) with regional lymph nodes metastasis. Serum sIL-2Ralpha concentration in 16 patients with malignant glioma varied greatly with the mean concentration of 328 +/- 192 U/ml. In 5 of 16 patients with malignant glioma, we could detect the significant increase of serum sIL-2Ralpha concentration from early stage of recurrence. CONCLUSIONS Serum levels of sIL-2Ralpha could be a useful immunological marker in patients with malignant brain tumors.

A stress-induced substance(s) (factor C) incubated with norepinephrine (NE) has strong flower-inducing activity in Lemna paucicostata. We isolated an essential component (FIF) of factor C, and clarified its chemical structure as 9-hydroxy-10-oxo-12(Z),15(Z)-octadecadienoic acid, an alpha-ketol derivative of linolenic acid, which is formed via 9-hydroperoxy linolenic acid. Synthesized FIF showed flower-inducing activity after incubation with NE (factor C activity) equivalent to that formed in the stressed Lemna. Jasmonic acid and 13-hydroxy-12-oxo-9(Z),15(Z)-octadecadienoic acid (12,13-alpha-ketol linolenic acid), both of which are formed via 13-hydroperoxide of linolenic acid and all other derivatives of FIF synthesized by chemical and enzymatic processes failed to show the factor C activity. These results suggest that the molecular structure of FIF is very specific for the factor C activity.

熱分解-ガスクロマトグラフ法 (Py-GC) に新規開発した液体試料用熱分解管を適用することによって, 前処理の簡略化および操作時間の短縮化が達成できた。Py-GCにそれぞれ電子イオン化質量分析計, 化学イオン化質量分析計, 原子発光検出器および赤外分光光度計を結合させたPy-GC-EIMS, Py-GC-CIMS, Py-GC-AEDおよびPy-GC-IR法を, ポリジメチルシロキサン (DMS) の熱分解生成物の同定に適用することにより, DMSとそれに含まれていた揮発性不純物の詳細な構造解析が可能となった。各検出法によって得られた同一熱分解物の保持時間には差異が認められたが, 標準ポリエチレンのパイログラムに出現する直鎖飽和炭化水素の保持時間で計算した保持指標 (RI) によって, その差異を補正することが可能となった。DMSの各検出法によるPy-GCで得られた五つのパイログラムに出現した主要な9本のピークについてRIを計算したところ, 同一ピークのRIにおけるバラツキを示す相対標準偏差 (RSD) は, いずれも0.6%以下であった。したがって, 各ピークの構造情報をRIを基準として集積させた統合ライブラリの構築が可能であると考えられた。また, Py-GC-AED法で得られた全炭素面積/全ケイ素面積比 (C/Si比) と核磁気共鳴スペクトル法から得られたM/Q比 (トリメチルシロキシケイ酸構造中のMおよびQ単位のモル比) との間には直線関係 (r=0.9997) が認められ, Py-GC-AEDによるM/Q比推定法を確立できる可能性を見いだした。しかしながら, 未知パイログラムの同定法を既存のEIMSライブラリ検索法によって確立することを試みたが, ポリアクリル酸の全イオンパイログラムから得られる合算EIMSスペクトルのライブラリ検索結果であるマッチクォリテイの再現性は, 期待した値よりもわずかに低く, 今後合算EIMSスペクトルの最適算出条件および検索パラメータの最適条件をさらに検討する必要があることがわかった。

Several recent reports have suggested that sperm count and quality in normal men are declining. Various environmental chemical compounds may affect the male reproductive system. We propose here that diesel exhaust is an environmental pollutant with the potential to influence male reproductive function. Ultrastructural changes were observed in Leydig cells of mice exposed to diesel exhaust (0.3 mg diesel exhaust particles (DEP)/ m 3 through the airway, 12 h daily, up to 6 months) and reduction in LH receptor mRNA expression in Leydig cells was observed at a concentration of 1 mg DEP/m 3 . Daily sperm production per gram of testis dose-dependently decreased with exposure to DE for 6 months; 29%, 36%, and 53% reductions were observed at 0.3, 1.0, and 3.0 mg DEP/m 3 , respectively. A no-observed-adverse-effect level (NOAEL) was observed with approximately 30 μg DEP/m 3 , which is lower than the WHO-recommended limit.

We examined the effect of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) on changes in gene expression of murine testicular Leydig TM3 cells by a method of differential mRNA display based on a PCR technique. Although no change in viable cell number was observed by TCDD at the concentration of 10nM, Ah receptor mRNA was down-regulated and CYP1A1 was induced in TM3 cells. One hundred and sixty-one PCR products, differentially expressed in 2% agarose gel electrophoresis after PCR amplification, were detected. Nine cDNAs were confirmed to be differentially expressed in Northern ELISA analysis. Two cDNAs were cloned and sequenced. One was identical to Homo sapiens Nedd-4-like ubiquitin-protein ligase WWP2 (WW domain-containing protein), which is associated with the ubiquitin proteolytic process. TCDD decreased expression of WWP2, which might cause the suppression of proteolytic process. The other was identical to the mouse expressed sequence tag. The function of this gene was not known. These genes might provide information for understanding TCDD-induced toxicity or biological responses.

We studied the effect of a streptococcal preparation, OK-432, on the cytotoxicity and the proliferation of lymphokine-activated killer (LAK) cells for use in adoptive immunotherapy. Peripheral blood mononuclear cells (PBMC) were derived from healthy donors and patients with malignant brain tumors. We divided PBMC into two groups; these cells then were stimulated with interleukin-2 in the presence or absence of OK-432. OK-432 was added only in the initial 3 days during the 3-week midterm culture period. Then, we compared OK-432-stimulated LAK (OK-LAK) cells with standard LAK (sLAK) cells in terms of their rate of proliferation and cytotoxicity. OK-LAK cells proliferated more rapidly than sLAK cells. The cytotoxicity of OK-LAK cells increased, whereas that of sLAK cells decreased. We also investigated the phenotypic differences between these two types of LAK cells and found that, on day 21, the OK-LAK cells consisted mostly of CD3 - CD56 + NK cells, whereas the sLAK cells consisted mostly of CD3 + CD56 - T cells. The difference in their level of cytotoxic potential might be explained by the difference of predominant phenotype.

Human glioma cell line, Onda 10 produces TGF-β1. TGF-β1 has a biological role for the immunosuppression of the host. We have investigated whether suppression of TGF-β1 on human glioma cell enhanced the susceptibility to lymphokine-activated killer (LAK) cells. In vitro, susceptibility to LAK cells on Onda 10 cell is augmented by retroviral gene transfection with antisense TGF-β1. Nude mice bearing Onda 10 cells transduced with antisense TGF-β1 gene has a longer life span compared to mice carrying that of sense TGF-β1 gene or vector alone. The cytotoxic activity of LAK cells induced from spleen cells of mice carrying antisense TGF-β1 gene transduced cells is higher against Onda 10 cell than that of LAK cells from mice carrying vector alone transduced cells. Also, antisense TGF-β1 gene transduced cells are much more sensitive to LAK cells compared to Onda 10. These suggest that the augmented host systemic immunity in mice is one of the mechanisms of the reduced tumorigenicity of antisense TGF-β1 gene transduced cells and that the increased systemic immunity could be ascribed to the increased immunogenicity of the tumor cells. The gene therapy for malignant glioma with antisense TGF-β1 gene is expected to be promising.

Background: Brain metastasis from esophageal carcinoma is rare, and its natural history is unclear. To determine predictors for their outcomes, we conducted a retrospective review of patients. Methods: We treated 803 patients with metastatic brain tumors, and there were 17 patients with brain metastases from esophageal carcinoma. Their median age at the diagnosis was 57 years. Results: In the 15 patients with the pretreatment KPS of 70 or higher, the median survival after the diagnosis of brain metastasis was 26.2 months. Seven patients showed the median survival of 17.7 months after resection alone. The median survival was 65.5 months in the 3 patients who were treated with resection plus radiation. Conclusions: Neurosurgical resection followed by WBRT seemed to be the indicated treatment in these patients.