Immunization of BALB/c mice with alum-adsorbed OVA, followed by three bronchoprovocations with aerosolized OVA, resulted in the development of airway hyperresponsiveness (AHR) and allergic inflammation in the lung accompanied by severe infiltration of eosinophils into airways. In this murine asthma model, administration of monoclonal anti-IL-5 Ab before each Ag challenge markedly inhibited airway eosinophilia, but the treatment did not affect the development of AHR. Immunization and aerosol challenges with OVA following the same protocol failed to induce AHR in the mast cell-deficient W/Wv mice, but induced AHR in their congenic littermates, i.e., WBB6F1 (+/+) mice. No significant difference was found between the W/Wv mice and +/+ mice with respect to the IgE and IgG1 anti-OVA Ab responses and to the airway eosinophilia after Ag provocations. It was also found that reconstitution of W/Wv mice with bone marrow-derived mast cells cultured from normal littermates restored the capacity of developing Ag-induced AHR, indicating that lack of mast cells was responsible for the failure of W/Wv mice to develop Ag-induced AHR under the experimental conditions. However, the OVA-immunized W/Wv mice developed AHR by increasing the frequency and Ag dose of bronchoprovocations. The results suggested that AHR could be developed by two distinct cellular mechanisms. One would go through mast cell activation and the other is IgE/mast cell independent but an eosinophil/IL-5-dependent mechanism.
Lymphoepithelioma-like carcinoma (LELC) is a tumor which occurs outside the nasopharynx and has morphological features identical to nasopharyngeal lymphoepithelioma. LELC of the breast (LELC-B) is uncommon, and its resemblance to medullary carcinoma of the breast (MC-B) obscures distinction between these two tumors. We report a case of LELC-B occurring in a 47-year-old woman. The tumor consisted of multinodules without circumscription. The tumor cells mainly exhibited loose clusters being permeated by numerous lymphocytes. The tumor cell clusters showed inconspicuous margins, which were far from syncytial patterns. The epithelial nature of the tumor cells was demonstrated by positivity for epithelial membrane antigen, AE1/AE3 and CAM5.2. Furthermore, glandular differentiation of the tumor cells was confirmed using electron microscopy for the first time. Epstein-Barr virus (EBV) was not detected using either in situ hybridization or polymerase chain reaction. These findings, together with former reports of LELC-B, suggest that the distinction between LELC-B and MC-B depends on whether circumscription and syncytial growth patterns exist. The other findings, including absence of EBV and immunohistochemical aspects of the tumor cells, are not considered different thus far. Although the prognosis of LELC-B is thought to be favorable, which is also similar to MC-B, distant metastasis was detected in the present case. To confirm the clinicopathological entity of these two tumors, it is important to recognize the difference between LELC-B and MC-B.
The enzyme activity for cytosine deaminase, which converts cytosine to uracil in bacterial, is usually undetected in higher plants and animals. The enzyme also catalyzes conversion of non-toxic 5-fluorocytosine (5-FC) to 5- fluorouracil (5-FU), a toxic compound for plant growth. The gene encoding cytosine deaminase (codA) from Escherichia coli was fused to cauliflower mosaic virus (CaMV) 35S promoter (P35S), and cloned into a binary vector pLABR101. The resulting plasmid pLABR102 contained two marker genes for plants: a positive marker gene, bialaphos resistance (bar) gene driven by the promoter from nopaline synthase gene (Pnos) and a negative one, P35S-codA. The binary vector pLABR102 was transformed into Arabidopsis thaliana via Agrobacterium-mediated transformation. In transgenic progenies (T3) of the second (T2) generation heterozygous for a single T-DNA insertion, a 3:1 segregation ratio was observed on both bialaphos (resistance to sensitive) and 5-FC (sensitive to unaffected). From T2 plants homozygous for the T-DNA insert, on the other hand, no segregation was detected: all the T3 seedlings were resistant to bialaphos and sensitive to 5-FC. PCR and Northern analyses showed that the 5-FC sensitivity in transgenic descendants was caused by the integration and expression of the chimeric codA gene in the Arabidopsis genome. The results indicated that cytosine deaminase from E. coli is functional and useful for negative selection in Arabidopsis, and that sensitivity to 5-FC as well as the positive bialaphos resistance are dominant traits in Arabidopsis.
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