RESUMO
Wild mouse feces can disseminate zoonotic microorganisms throughout a farm, which is a great threat to human health and can lead to economic loss through contaminated agricultural produce. To assess the microbial communities, especially fecal coliform bacteria, we used two methods. First, we isolated bacterial colonies onto the common media LB (lactose broth) agar, TSA (tryptic soy agar), and MRS (de Man, Rogosa, and Sharpe) agar, and then randomly select colonies from each plate and stocked them to the mother plate for genomic DNA isolation. Second, we analyzed bacterial colonies using the 16S rRNA gene molecular diagnostic method. Based on bacterial cultures and bacterial 16S rRNA gene markers, we detected four different bacterial species (Bacillus amyloliquefaciens, Escherichia coli, Staphylococcus xylosus, and Serratia liquefaciens) from fecal coliforms of the striped field mouse Apodemus agrarius and A. peninsulae in agricultural areas in South Korea. These results could help us to better understand the pathogen reservoirs of mice and initiate some preventive measures to mitigate the microbial risks associated with mouse fecal matter in agricultural production areas.
Assuntos
Microbiota , Murinae/microbiologia , Animais , Bacillus amyloliquefaciens/genética , Bacillus amyloliquefaciens/isolamento & purificação , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Fezes/microbiologia , RNA Ribossômico 16S/genética , República da Coreia , Serratia liquefaciens/genética , Serratia liquefaciens/isolamento & purificação , Staphylococcus/genética , Staphylococcus/isolamento & purificaçãoRESUMO
PURPOSE: The KIT inhibitor, imatinib, has shown promising efficacy in patients with KIT-mutated melanoma; however, acquisition of resistance to imatinib occurs rapidly in the majority of patients. The mechanisms of acquired resistance to imatinib in melanoma remain unclear. METHODS: We analyzed biopsy samples from paired baseline and post-treatment tumor lesions in one patient with KIT-mutated melanoma who had had an initial objective tumor regression in response to imatinib treatment followed by disease progression 8 months later. RESULTS: Targeted deep sequencing from post-treatment biopsy samples detected an additional mutation in CTNNB1 (S33C) with original KIT L576P mutation. We examined the functional role of the additional CTNNB1 S33C mutation in resistance to imatinib indirectly using the Ba/F3 cell model. Ba/F3 cell lines transfected with both the L576P KIT mutation and the CTNNB1 S33C mutation demonstrated no growth inhibition despite imatinib treatment, whereas growth inhibition was observed in the Ba/F3 cell line transfected with the L576 KIT mutation alone. CONCLUSIONS: We report the first identification of the emergence of a CTNNB1 mutation that can confer acquired resistance to imatinib. Further investigation into the causes of acquired resistance to imatinib will be essential to improve the prognosis for patients with KIT-mutated melanoma.
Assuntos
Resistencia a Medicamentos Antineoplásicos/genética , Mesilato de Imatinib/farmacologia , Neoplasias Pulmonares/genética , Melanoma/genética , Mutação , Neoplasias Cutâneas/genética , beta Catenina/genética , Proliferação de Células/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/secundário , Masculino , Melanoma/tratamento farmacológico , Melanoma/patologia , Pessoa de Meia-Idade , Prognóstico , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-kit , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/secundário , Células Tumorais CultivadasRESUMO
PURPOSE: To evaluate the treatment outcomes of low-dose whole brain radiation therapy (WBRT)-based differential radiation therapy (RT) for metastatic brain tumors. METHODS: A total of 242 targets (metastatic brain lesions) were analyzed in the present study. Median WBRT dose and number of fractions were 25 (range 25-35) Gy and 10 (range 8-15) fractions, respectively. A median normalized total dose (NTD) of 1.8 Gy (NTD(1.8Gy)) to the metastatic lesion was 45 (range 27-64.8) Gy. We numbered and contoured each metastatic lesion sequentially using computed tomography fused with serial magnetic resonance imaging to evaluate volumetric changes. RESULTS: The 6-month and 1-year freedom from remote intracranial failure rates were 87.7 and 58.5 %, respectively. The 6-month actuarial local control (LC) rate was 93.4 %. Tumor diameter was a major determinant for LC, and tumor histology was a significant parameter predicting the volume reduction rate. With overall complete response (CR) rate of 56.6 % after RT, CR rate, if the target was more than 1 cm in size, was 25 % with a median NTD(1.8Gy) of 45 Gy, requiring dose escalation to achieve better target regression. CONCLUSIONS: Low-dose WBRT with selective boost was feasible and effective. Our results pose the rationale of future trial of differential radiation therapy (RT), which prescribes different radiation dose according to the tumor density in metastatic brain tumors.
Assuntos
Neoplasias Encefálicas/radioterapia , Neoplasias Encefálicas/secundário , Irradiação Craniana , Fracionamento da Dose de Radiação , Neoplasias/radioterapia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Encefálicas/mortalidade , Estudos de Viabilidade , Feminino , Seguimentos , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Neoplasias/mortalidade , Neoplasias/patologia , Estudos Retrospectivos , Taxa de Sobrevida , Tomografia Computadorizada por Raios X , Resultado do Tratamento , Adulto JovemRESUMO
Preimplantation genetic diagnosis (PGD) has become an assisted reproductive technique for couples that have genetic risks. Despite the many advantages provided by PGD, there are several problems, including amplification failure, allele drop-out and amplification inefficiency. We evaluated multiple displacement amplification (MDA) for PGD of the fragile X syndrome. Whole genome amplification was performed using MDA. MDA products were subjected to fluorescent PCR of fragile X mental retardation-1 (FMR1) CGG repeats, amelogenin and two polymorphic markers. In the pre-clinical tests, the amplification rates of the FMR1 CGG repeat, DXS1215 and FRAXAC1 were 84.2, 87.5 and 75.0%, respectively, while the allele dropout rates were 31.3, 57.1 and 50.0%, respectively. In two PGD treatment cycles, 20 embryos among 30 embryos were successfully diagnosed as 10 normal embryos, four mutated embryos and six heterozygous carriers. Three healthy embryos were transferred to the uterus; however, no clinical pregnancy was achieved. Our data indicate that MDA and fluorescent PCR with four loci can be successfully applied to PGD for fragile X syndrome. Advanced methods for amplification of minuscule amounts of DNA could improve the sensitivity and reliability of PGD for complicated single gene disorders.
Assuntos
Proteína do X Frágil da Deficiência Intelectual/genética , Síndrome do Cromossomo X Frágil/diagnóstico , Síndrome do Cromossomo X Frágil/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Diagnóstico Pré-Implantação/métodos , Repetições de Trinucleotídeos/genética , Adulto , Feminino , Humanos , Masculino , GravidezRESUMO
This study was conducted to evaluate 1) the effects of soybean meal (SBM) from 3 major SBM-producing countries (United States, Brazil, and India) on the growth performance, nutrient digestibility, and meat quality of growing-finishing pigs and 2) the feeding value of the evaluated SBM as a feedstuff for growing-finishing pigs. Soybean meal representative of each country was purchased on the open market. A total of 144 barrows with the average BW of 23.77 (SD = 1.52 kg) were allotted to 3 dietary treatments with 12 replicate pens per treatment and 4 pigs per pen. The US SBM used in this trial had greater CP and total AA contents, greater KOH protein solubility, and less crude fiber than SBM from either Brazil or India. Diets were formulated to be isolysinic and isocaloric. Pigs fed diets containing US SBM were heavier (P < 0.05) than pigs fed diets containing Brazilian or Indian SBM at 12 and 18 wk. Pigs fed diets containing US SBM had greater (P < 0.05) ADG and G:F throughout the overall period when compared with pigs fed SBM produced in Brazil or India. The DM and N digestibility were greater for US SBM (P < 0.05) than Brazilian and India SBM at 6, 12, and 18 wk. Pigs were slaughtered at the end of 18 wk, and no differences were observed on meat quality (backfat thickness, LM area, pH, meat color, water holding capacity, and drip loss) among the treatments, except lean percentages were less and backfat thickness was greater in pigs fed Brazilian and Indian SBM than US SBM. The cost per kilogram of BW gain of the experimental diet containing US SBM was less than that of diets containing Brazilian or Indian SBM, but it would depend on the relative prices of each SBM. Based on the results of the current experiment, the productivity and feeding value of US SBM seems to be greater than the SBM produced in Brazil and India.