Fungi of the Metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. To establish a co-transformation system for the transformation of multiple M. robertsii genes using Agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. Subsequently, in the two vectors containing the nourseothricin and phosphinothricin resistance cassettes were inserted eGFP and mCherry expression cassettes, respectively. These new vectors were then introduced independently into A. tumefaciens and used to transform M. robertsii either in independent events or in one single co-transformation event using an equimolar mixture of A. tumefaciens cultures. The number of transformants obtained by co-transformation was similar to that obtained by the individual transformation events. This method provides an additional strategy for the simultaneous insertion of multiple genes into M. robertsii.
Agrobacterium ; Agrobacterium tumefaciens ; Fungi ; Insects ; Metarhizium* ; Methods ; Streptothricins ; Virulence

OBJECTIVE: The objective of study is to evaluate the incidence of leptomeningeal carcinomatosis (LMC) in breast cancer patients with parenchymal brain metastases (PBM) and clinical risk factors for the development of LMC. METHODS: We retrospectively analyzed 27 patients who had undergone surgical resection (SR) and 156 patients with whole brain radiation therapy (WBRT) as an initial treatment for their PBM from breast cancer in our institution and compared the difference of incidence of LMC according to clinical factors. The diagnosis of LMC was made by cerebrospinal fluid cytology and/or magnetic resonance imaging. RESULTS: A total of 27 patients (14%) in the study population developed LMC at a median of 6.0 months (range, 1.0-50). Ten of 27 patients (37%) developed LMC after SR, whereas 17 of 156 (11%) patients who received WBRT were diagnosed with LMC after the index procedure. The incidence of LMC was significantly higher in the SR group compared with the WBRT group and the hazard ratio was 2.95 (95% confidence interval; 1.33-6.54, p<0.01). Three additional factors were identified in the multivariable analysis : the younger age group (<40 years old), the progressing systemic disease showed significantly increased incidence of LMC, whereas the adjuvant chemotherapy reduce the incidence. CONCLUSION: There is an increased risk of LMC after SR for PBM from breast cancer compared with WBRT. The young age (<40) and systemic burden of cancer in terms of progressing systemic disease without adjuvant chemotherapy could be additional risk factors for the development of LMC.
Brain ; Breast ; Breast Neoplasms ; Chemotherapy, Adjuvant ; Humans ; Incidence ; Magnetic Resonance Spectroscopy ; Meningeal Carcinomatosis ; Neoplasm Metastasis ; Retrospective Studies ; Risk Factors ; Streptothricins

BACKGROUND: Serologic tests for specific antibody nowadays are widely employed for the diagnosis of parasitic diseases. Recently, an increasing numbers of kits have adopted enzyme-linked immunosorbent assay (ELISA) for the detection of parasitic antibodies. In this study, we evaluated two ELISA reagents for the diagnosis of parasitic diseases. METHODS: A total of 553 serum and 156 CSF samples were assayed using an in-house micro-ELISA and Genedia(R) Ab ELISA (Green cross PBM, Korea) for Cysticercus, Paragonimus westermani, Clonorchis sinensis, and Sparganum. We reviewed the medical records of all patients. The results from Genedia(R) Ab ELISA kit were compared with those from the in- house micro-ELISA method. RESULTS: The overall concordance rate between the two ELISA tests was 95.5%. When compared with the clinical information, the sensitivity, specificity, positive predictive value, and negative predictive value of the in-house micro-ELISA were 100%, 99.0~99.6%, 82.4~96.4%, and 100%, and the respective figures for Genedia(R) Ab ELISA kit were 92.9~100%, 88.0~97.3%, 41.7~50%, and 99.9~100% with kappa agreement of 0.53-0.63. Comparison of two ELISA methods showed a significant difference (P<0.05). Retesting of 85 discordant samples showed that the concordance rate of the in-house ELISA was 97.7% and that of Genedia(R) Ab ELISA was 28.2%. CONCLUSION: Genedia(R) Ab ELISA kit showed an intermediate level of kappa agreement compared with the in-house ELISA. Further studies are necessary to improve the concordance rate of the two methods, and a careful interpretation of these results is required for a precise diagnosis.
Antibodies ; Clonorchis sinensis ; Cysticercus ; Enzyme-Linked Immunosorbent Assay ; Humans ; Indicators and Reagents ; Medical Records ; Paragonimus westermani ; Parasitic Diseases ; Sensitivity and Specificity ; Serologic Tests ; Sparganum ; Streptothricins