Conversion of Normal Ly-1-Positive B-Lineage Cells into Ly-1-Positive Macrophages in Long-Term Bone Marrow Cultures

We obtained eight different cell lines in the long-term bone marrow culture system that showed a germ-line configuration of the joining (J) region segments of the Ig heavy-chain (IgH) genes. Their surface markers were CD45R+, Ly-1+, Lyb-2+, cIgM-, sIgM-, Ia-, Thy-1-, Mac-1-, and IL-2R (Tac)+. Use of very young mice and the presence of IL-5 were important for preferential promotion of the survival of B-lineage lymphocytes bearing the Ly-1 markers. When we treated two of them (J8 and J10) with 5-azacytidine for 24 h followed by co-culture with stromal cells and IL-.5, they became Ly-1+, sIgM+ B cells, and Ly-1+, Mac-1+ macrophagelike cells, respectively. After other early lymphoid lines (J1, J8, and J13) were maintained by co-culture with ST2 and IL-5 for more than a year, they showed a heterogeneous DNA rearrangement profile of the J region segment of the IgH gene, although only J13 rearranged the κ-light chain gene. Northern blot analysis revealed that these cell lines expressed Cμ-mRNA, and λ5-mRNA, consistent with normal pre-B cells. Intriguingly, J1, J8, and J13 expressed c-fms mRNA constitutively. When J13 cells were co-cultured with ST2 and GM-CSF in place of ST2 and IL-5, they acquired Mac-1 expression and retained Ly-1 expression. They were morphologically macrophages, nonspecific-esterase-positive, and showed phagocytosis of latex beads. These results support evidence for a close relationship between the myeloid and Ly-1+ B-cell pathways of differentiation, and indicate that our IL- 5-dependent clones are multipotential intermediates in differentiation from pro-B cells to B cells and macrophages

Development of two types of mite-allergen induced murine models of chronic asthma with different severity

Asthma is an allergic disease characterized by chronic airway inflammation, hyper-responsiveness (AHR), and reversible obstruction. The main inflammatory changes are induced by infiltration of eosinophils into the airway. Few animal models resemble the spontaneous history of asthma due to variations in the selection of the mouse strain, appropriate antigen, and exposure methods. Here, we prepared two different mouse models in which the mechanism was close to that of human asthma. We transnasally administered mite Dermatophagoides farinae (Df) allergen to BALB/c mice 10 times (Df-2) or 25 times (Df-5). After comparison with mice administered phosphate-buffered saline, the AHR and immediate asthmatic response were evaluated, in addition to the number of eosinophils in the bronchoalveolar lavage fluid (BALF). Df-specific IgE and IgG1 levels in the serum, and Th2 cytokines (interleukin [IL]-5, IL-13) in the BALF were measured by enzymelinked immunosorbent assay. Immediate asthmatic response and AHR were enhanced in mite allergen-treated mice (Df-2 and Df-5) compared to PBS-treated mice. The number of eosinophils and IL-13 levels in the BALF, and specific IgE in the serum were greater in Df-5 than in Df-2 mice. We established two different murine chronic asthma models, in which the severity depended on the number of exposures to Df. Greater intranasal exposure to a Df allergen resulted in more severe asthma in a BALB/c mouse model

Response of epilithic diatom communities to environmental gradients in subtropical temperate Brazilian rivers

This work aims to analyse the response of epilithic diatom communities to enviranmental gradients in subtropical temperate southern Brazilian rivers to contribute to the development of a widely applicable methodology for water-quality monitoring. Samples for physical, chemical and biological determination were collected monthly, fram December 2001 to November 2002 and fram March 2003 to February 2004, at 9 stations along the rivers Pardo and Pardinho in the hydrographical basin of Rio Pardo, State of Rio Grande do Sul (RS), Brazil. Physical and chemical variables (water temperature, pH, dissolved oxygen, biochemical oxygen demand, chemical oxygen demand, nitrates, nitrites, phosphates and total dissolved solids) and biological variables (epilithic diatom communities) were used as parameters for water-quality assessment. The data matrix was examined by means of a multivariate ordination using Detrended Correspondence Analysis (DCA). In total, 270 taxa were identified to specific or infra-specific levels. The results showed that of all the physical and chemical variables used, the main gradient along the DCA first axis was eutraphication, which was indicated by its significant correlation with phosphates (p < 0.00l). Hence, the species' scores on the DCA axis were used as an operational criterion for indicating their tolerance to eutrophication. From this analysis, the 10 taxa that were the most tolerant to eutrophication were: Cyclotella meneghiniana, Fallacia monoculata, Nitzschia acicularis, N. clausii, N. nana, N. palea, Nitzschia sp., Pinnularia sp., Sellaphora pupula sensu lato and Ulnaria acuso The present study showed that epilithic diatom assemblages reflect anthropogenic changes in hydrographic basins in subtropical temperate streams, especially pollution by organic enrichment and eutraphication. Key words: Diatoms, eutrophication, biomonitoring, southern Brazilian rivers.Este trabajo pretende analizar la respuesta de la comunidad de diatomeas epilíticas a gradientes ambientales en ríos brasileños templados subtropicales, con el objetivo de contribuir al desarrollo de una metodología ampliamente aplicable para la monitorización de la calidad del agua. Diversas muestras para determinaciones físicas, químicas y biológicas fueron recogidas mensualmente, de diciembre de 2001 a noviembre de 2002, y de marzo de 2003 afebrero de 2004, en nueve esta­ciones de muestre o distribuidas a lo largo de los ríos Pardo y Pardinho, localizados en la Cuenca Hidrográfica del Río Pardo, Estado del Río Grande del Sur; Brasil. Variables físicas y químicas (temperatura del agua, pH, oxígeno disuelto, demanda bioquímica de oxígeno, demanda química de oxígeno, nitratos, nitritos, fosfato y sólidos totales disueltos) y biológicas (co­munidad de diatomeas epilíticas) fueron usadas como parámetros para evaluar la calidad del agua. El conjunto de datos fue analizado por medio de una ordenación multivariante usando el Análisis de Correspondencias sin Tendencias (DCA). En total, 270 taxones fueron identificados a nivel especifico o infra-especifico. Los resultados indicaron que, entre todas las variables físicas y químicas usadas, el principal gradiente establecido a lo largo del primer eje DCA fue la eutrojización, señalada por su correlación significativa con la concentración de fosfatos (p < 0.001). De esta forma, los valores alcanzados por las especies en el primer eje DCA fueron usados como un criterio operativo para indicar su tolerancia a la eutrojización. Los 10 taxones más tolerantes a la eutrojizaciónfueron: Cyc10tella meneghiniana, Fallacia monoculata, Nitzschia acicularis, N. c1ausii, N. nana, N. palea, Nitzschia sp., Pinnularia sp., Sellaphora pupula sensu lato y Ulnaria acuso El presente estudio muestra que la comunidad de diatomeas epilíticas refleja cambios antropogénicos en ríos templados subtropicales de cuencas hidrográjicas, especialmente la contaminación por enriquecimiento orgánico y eutrofización

Response of epilithic diatom communities to environmental gradients in subtropical temperate Brazilian rivers

P. 323-340This work aims to analyse the response of epilithic diatom communities to environmental gradients in subtropical temperate southern Brazilian rivers to contribute to the development of a widely applicable methodology for water-quality monitoring. Samples for physical, chemical and biological determination were collected monthly, from December 2001 to November 2002 and from March 2003 to February 2004, at 9 stations along the rivers Pardo and Pardinho in the hydrographical basin of Rio Pardo, State of Rio Grande do Sul (RS), Brazil. Physical and chemical variables (water temperature, pH, dissolved oxygen, biochemical oxygen demand, chemical oxygen demand, nitrates, nitrites, phosphates and total dissolved solids) and biological variables (epilithic diatom communities) were used as parameters for water-quality assessment. The data matrix was examined by means of a multivariate ordination using Detrended Correspondence Analysis (DCA). In total, 270 taxa were identied to specic or infra-specic levels. The results showed that of all the physical and chemical variables used, the main gradient along the DCA rst axis was eutrophication, which was indicated by its signicant correlation with phosphates ( p < 0.001). Hence, the species’ scores on the DCA axis were used as an operational criterion for indicating their tolerance to eutrophication. From this analysis, the 10 taxa that were the most tolerant to eutrophication were: Cyclotella meneghiniana, Fallacia monoculata, Nitzschia acicularis, N. clausii, N. nana, N. palea, Nitzschia sp., Pinnularia sp., Sellaphora pupula sensu lato and Ulnaria acus. The present study showed that epilithic diatom assemblages reect anthropogenic changes in hydrographic basins in subtropical temperate streams, especially pollution by organic enrichment and eutrophicationS

Establishment of an asthma model by sensitization with mite antigen alone in C57BL/6J mice

Bronchial asthma is characterized by the bronchial hyperresponsiveness and airway obstruction related to airway smooth muscle contraction. Eosinophilic airway inflammation is involved in its pathogenesis. To reproduce the condition, various animal models have been prepared. However, there are many models that do not reflect the spontaneous history of bronchial asthma onset in humans due to the mouse strain, sensitizing antigen, or administration method. In this study, we prepared a mouse model of which the mechanism is similar to that of human bronchial asthma. Mite Extract-Dermatophagoides farinae (Derf) antigen was transnasally administered to wild-type C57BL/6J mice (WT) 13 times. Subsequently, an airway hypersensitivity test (Mch PC_200), specific antigen exposure test (ΔSRaw), bronchoalveolar lavage (BAL), and blood collection were performed to examine the presence or absence of asthma acquisition and differences in the local pulmonary levels of cytokines/chemokines in comparison with the physiological saline-treated group. In the mite antigen-treated mice (WT/-Derf), bronchial hyperresponsiveness was enhanced, antigen-specific was increased airway resistance in comparison with physiological salinetreated mice (WT/-Saline). In addition, the number of eosinophils in BAL fluid (BALF) was greater. Furthermore, there was a correlation among leukotrienes, eotaxin, and tissue inhibitors of metalloproteinase 1 in BALF, suggesting that the mechanism concerning eosinophilic airway inflammation involving in human bronchial asthma was reproduced. In this study, we successfully established a mouse bronchial asthma model in which the pathogenesis resembles that in humans in comparison with conventional models, using Derf antigen alone and C57BL/6J mice

免疫抑制剤投与中に急性呼吸促迫症候群を合併した粟粒結核の１例

症例は75歳，女性．MPO-ANCA 関連血管炎に対して半年間，ステロイド薬が投与されていた．４日前から発熱，呼吸困難が出現，意識障害も伴ってきたため，当院を受診した．画像上，両側肺にびまん性の中枢側に有意な浸潤影とすりガラス陰影を認め，急性呼吸促迫症候群（ARDS）の合併を疑われた．人工呼吸管理となり，挿管中に採取した喀痰抗酸菌検査で塗抹陽性，結核菌PCR陽性の結果が得られ，血液や尿からも結核菌が検出され，粟粒結核によるARDS と診断した．治療は入院後の第３病日からINH ＋ RFP ＋ EB による抗結核療法を開始し，人工呼吸管理および血液透析をしながら経過観察をしていたが，播種性血管内凝固症候群も併発し，第14病日に死亡した．ARDS を合併する粟粒結核の症例も散見されることから，鑑別診断に粟粒結核も念頭におきながら診療することが重要と思われた．A 75-year-old woman, who was treated with corticosteroid therapy for six months for MPO-ANCA related vasculitis, visited to our hospital with fever, dyspnea and consciousness disturbance four days ago. She was diagnosed with acute respiratory distress syndrome (ARDS) from radiological findings such as diffuse bilateral hilar dominant infiltration shadow and ground-glass opacity. She was admitted to the intensive care unit (ICU) and put on mechanical ventilation for acute respiratory failure. The aspiration sputum after incubation turned out to be positive for acid-fast bacilli, which were identified as Mycobacterium tuberculosis (MTB) using a polymerase chain reaction test. As the same results in the clinical specimens of peripheral blood and urine were obtained, we made final diagnosis of miliary tuberculosis complicated with ARDS. Although we initiated the anti-tuberculosis treatment using INH, RFP, EB with mechanical ventilation and hemodialysis treatment, she died of multiple organ failure complicated with disseminated intravascular coagulation (DIC). Due to the fact, that we have encountered few cases of miliary tuberculosis complicated with ARDS, it is important that we suspect severe miliary tuberculosis in patients with immunosuppressive treatment