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@#Abstract: Objective To establish a rapid detection assay based on fluorescence recombinase polymerase amplification (RPA) targeting Necator americanus eggs, and to evaluate its efficacy, providing technical support for rapid detection of Necator americanus in fecal samples. Methods The fluorescence RPA primers and probe were designed based on the cox1 gene of Necator americanus and then screened the optimal combination to develop the assay. The genomic DNA of Necator americanus eggs was diluted to 7 concentration gradients including 100 pg/µL, 10 pg/µL, 1 pg/µL, 100 fg/µL, 10 fg/µL, 1 fg/µL, 0.1 fg/µL, to determine the detection limit of the assay. The specificity of the assay was demonstrated by detected genomic DNA from Schistosoma japonicum, Ascaris lumbricoides, Clonorchis sinensis and Fasciola hepatica. A total of 44 fecal samples were collected and DNA extraction was performed, and the modified Kato-Katz method, semi-nest PCR method, and fluorescent RPA method were simultaneously used for detection to evaluate the sensitivity and specificity. Results The established fluorescence RPA assay can specifically amplify a fragment of 194 bp of the Necator americanus cox1 gene within 20 min, with a detection limit of 10 fg/µL. There was no cross-reactivity with Schistosoma japonicum, Ascaris lumbricoides, Clonorchis sinensis, Fasciola hepatica after specificity validation. In 44 fecal samples, 27 positive samples were detected by the fluorescence RPA assay, and 26 positive samples were detected by both the Kato-Katz and the semi-nested PCR. The fluorescence curve of sample number 1 was slightly higher than the negative control in the later stage of the reaction, but did not show a similar trend to the positive control, and was therefore judged to be a suspected negative sample. Compared with the Kato-Katz method and the semi-nest PCR method, The sensitivity of the fluorescent RPA method were 100.00% and the specificity were 94.44%, and the consistency of the detection results was good (Kappa=0.953>0.75). Conclusions The assay based on the fluorescence RPA is an efficient, sensitive and specific technique for detecting Necator americanus and it can be applied for surveillance and early warning of hookworm infection.
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Objective:To investigate the regulating molecules and acting mechanism of TAB182 in HR pathway.Methods:TAB182 in human breast cancer MCF-7 cells was knocked down by shRNA strategy, the TAB182 knockdown MCF-7 as the TAB182 knockdown group, and the MCF-7 cell using the shRNA negative control as the TAB182 negative control group. RNA sequencing and qRT-PCR were performed to screen and verify the differentially expressed genes of HR pathway related to TAB182 depression. Western blot was used to detect protein expression. Immunofluorescence staining of nuclear RAD51 and BrdU was used to check the 3′ ssDNA formation by the end resection. The cell cycle arrest and apoptosis were measured by flow cytometry. Cloning formation assay was used to evaluate the sensitivity TAB182-knockdown cells to radiation.Results:Both quantitative RNA sequencing and qRT-PCR assays showed that TAB182-knockdown significantly decreased the mRNA expression of RPA2( t=17.97, P<0.05). Compared with the TAB182 negative control group, the protein level of RPA2, the number of RAD51 foci, and the 3′ ssDNA-binding nuclear protein marker BrdU in TAB182-knockdown cells were significantly reduced. At 4, 8, and 12 h after actinomycin D treatment, the attenuation of RPA2 mRNA in the TAB182-knockdown cells was accelerated ( t=5.37, 3.79, 3.69, P<0.05). Compared with the TAB182 negative control group, the radiosensitivity and radiation-induced apoptosis in the TAB182-knockdown group were increased ( t=3.48, 11.05, P<0.05), and at 24 h after irradiation, the cell cycle block time was prolonged ( t=8.40, P<0.01). Conclusions:TAB182 plays a role in maintaining RPA2 mRNA stability, thereby promoting HR repair. TAB182 knockdown cells are highly sensitive to ionizing radiation.
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The study evaluated the efficacy and soybean spectral responses to fifteen foliar fungicide mixtures labeled to control Asian soybean rust. Canopy level reflectance was measured using a multispectral camera onboard a multirotor drone before and two hours after each spray. The third application of fungicides improved control of soybean rust and increased yield. Nevertheless, up to three consecutive foliar fungicides applications did not affect the reflectance of soybean plants at visible and infrared wavelengths. Thus, drones can be a viable strategy for data acquisition regardless of the application of the fungicides.
Esse estudo avaliou a eficácia e as respostas espectrais de plantas de soja a quinze misturas de fungicidas utilizados no controle da ferrugem asiática da soja (FAS). A refletância do nível do dossel foi medida usando uma câmera multiespectral a bordo de um drone multirotor antes e duas horas após cada pulverização. A terceira aplicação de fungicidas melhorou o controle de FAS e aumentou a produtividade. Porém, três aplicações foliares consecutivas de fungicidas não afetaram a refletância de plantas de soja nos comprimentos de onda visível e infravermelho. Assim, drones podem ser uma estratégia viável para aquisição de dados independentemente da aplicação de fungicidas.
Subject(s)
Soybeans/physiology , Fungicides, Industrial/administration & dosage , Fungicides, Industrial/analysis , Sustainable Agriculture , Hyperspectral Imaging/methodsABSTRACT
Background: Whole brain radiotherapy for patients with brain metastasis from lung cancer – prognostic significance of RTOG-RPA score. Aim: To evaluate the prognostic significance of Recursive partition Analysis (RPA) score in predicting the survival in patients with brain metastasis from Non-Small Cell Lung Cancer (NSCLC). Materials and methods: 35 patients diagnosed to have brain metastasis (BM) from primary NSCLC who had received palliative whole brain radiotherapy (WBRT) with or without chemotherapy from March 2014 to Mar 2017 were analyzed in this study. Data regarding the patient age, gender, performance status, histology, number of BM, time of metastasis, neurosurgical resection, radiotherapy details were collected and analyzed. Patients were divided into 3 groups according to RPA classification. The differences in clinical characteristics and treatment variables were analyzed by chi square test and overall survival analysis using Kaplan Mayer. The Cox proportional hazards regression model was used to determine statistically significant variables related to survival. Results: In univariate analysis histology, number of BM, extra cranial metastases, KPS and RPA score were identified to have prognostic significance. The result of multivariate analysis by the Cox proportional hazard model showed that RPA, no of mets and Extra cranial mets were significant. S. Jeeva, K. Chandralekha, V. Vanitha, M. Sornam, Balasubramanium, P. Vidya. Whole brain radiotherapy for patients with brain metastasis from lung cancer – Prognostic significance of RTOG-RPA score. IAIM, 2019; 6(5): 32-39. Page 33 Conclusions: Our study showed that RPA is good prognostic indicator in assessing the prognosis of patients with brain metastasis in NSCLC.
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OBJECTIVE: The authors analyzed the effectiveness and therapeutic response of Novalis shaped beam radiosurgery for metastatic brain tumors, and the prognostic factors which influenced the outcome. METHODS: We performed a retrospective analysis of 106 patients who underwent 159 treatments for 640 metastatic brain lesions between January 2000 and April 2008. The pathologies of the primary tumor were mainly lung (45.3%), breast (18.2%) and GI tract (13.2%). We classified the patients using Radiation Therapy Oncology Group Recursive Partitioning Analysis (RPA) and then analyzed the survival and prognostic factors according to the Kaplan Meier method and univariate analysis. RESULTS: The overall median actuarial survival rate was 7.3 months from the time of first radiosurgery treatment while 1 and 2 year actuarial survival estimates were 31% and 14.4%, respectively. Median actuarial survival rates for RPA classes I, II, and III were 31.3 months, 7.5 months and 1.7 months, respectively. Patients' life spans, higher Karnofsky performance scores and age correlated closely with RPA classes. However, sex and the number of lesions were not found to be significantly associated with length of survival. CONCLUSION: This result suggests that Novalis radiosurgery can be a good treatment option for treatment of the patients with brain metastases.
Subject(s)
Humans , Brain , Brain Neoplasms , Breast , Gastrointestinal Tract , Lung , Neoplasm Metastasis , Radiosurgery , Retrospective Studies , Survival RateABSTRACT
Using uniform design to optimize some relative factors influencing the expressing of t-PA variant-rPA(K) in E.coli system, and found the best expressive conditions. They were(by using 300ml culture fask): the volume of media was 25ml, inducing time was 5.3 hours, pH was 6.0, the concentration of IPTG was 0.1mmol/L, inducing time was 25℃, and best culture media was HD. After being optimized, the yield of expression had been improved from 0.16 to 0.48, and it was as 3 times as before. The results above will offer the basement for purification and renaturation of rPA(K).
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OBJECTIVE: This study is performed to evaluate the role of gamma knife radiosurgery(GKS) for treatment of patients with large number of metastatic brain lesions. METHODS: The authors retrospectively reviewed the clinical outcome of 99 patients who underwent 124 GKS procedures for a total of 564 metastatic lesions. Sixty-three patients presented with 1 to 3 lesions (group A) and 36 patients presented with 4 or more lesions (group B) at initial GKS. The authors assessed survival according to a variety of possible prognostic factors. RESULTS: Median survival from the GKS for all cases was 28 weeks. The recursive partitioning analysis (RPA) class (p<0.0001) and the total tumor volume (p=0.0174) were significant factors influencing survival. The number of lesions, whole brain radiotherapy, primary tumor site, age, and sex were not significant. Median survival of group A (28 weeks) was not different from that of group B (28 weeks) (p=0.4027). Median survival was 66, 28, and 13 weeks for RPA class I, II, and III respectively in group A and 44, 36, and 11 weeks in group B. CONCLUSION: It is suggested that GKS provides a survival gain even in patients with a large number (4 or more) of metastatic lesions. Concerning the survival and the selection of patients for radiosurgery, RPA class and the total tumor volume should be considered as more important factors than the number of lesions. Multiplicity of the lesions alone should not be a factor for contraindication of radiosurgery.
Subject(s)
Humans , Brain , Neoplasm Metastasis , Radiosurgery , Radiotherapy , Retrospective Studies , Tumor BurdenABSTRACT
Ojective To research the appropriate routes the clinical results of the three different procedures for the laparoscopic adrenalectomy routes were compared. Methods Eighty six patients underwent laparoscopic adrenalectomy through three different routes-transperitoneal anterior (TAA), retroperitoneal lateral flank(RLA) and retroperitoneal posterior lumbar approaches(RPA). Results The time for performing operation through RPA and RLA was shorter than that through TAA(P