1.Development of a real-time SYBR Green PCR assay for the rapid detection of Dermatophilus congolensis.
Alfredo GARCIA ; Remigio MARTINEZ ; Jose Manuel BENITEZ-MEDINA ; David RISCO ; Waldo Luis GARCIA ; Joaquin REY ; Juan Manuel ALONSO ; Javier Hermoso DE MENDOZA
Journal of Veterinary Science 2013;14(4):491-494
Methods such as real time (RT)-PCR have not been developed for the rapid detection and diagnosis of Dermatophilus (D.) congolensis infection. In the present study, a D. congolensis-specific SYBR Green RT-PCR assay was evaluated. The detection limit of the RT-PCR assay was 1 pg of DNA per PCR reaction. No cross-reaction with nucleic acids extracted from Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus, or Austwickia chelonae was observed. Finally, the RT-PCR assay was used to evaluate clinical samples collected from naturally infected animals with D. congolensis. The results showed that this assay is a fast and reliable method for diagnosing dermatophilosis.
Actinomycetales/*isolation & purification
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Actinomycetales Infections/diagnosis/microbiology/*veterinary
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Animals
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Cattle
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Cattle Diseases/*diagnosis/microbiology
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Fluorescent Dyes/*diagnostic use
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Horse Diseases/*diagnosis/microbiology
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Horses
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Limit of Detection
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Real-Time Polymerase Chain Reaction/*methods/veterinary
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Reproducibility of Results
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Sheep
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Sheep Diseases/*diagnosis/microbiology
2.The NER-related gene GTF2H5 predicts survival in high-grade serous ovarian cancer patients.
Javier GAYARRE ; Marta M KAMIENIAK ; Alicia CAZORLA-JIMENEZ ; Ivan MUNOZ-REPETO ; Salud BORREGO ; Jesus GARCIA-DONAS ; Susana HERNANDO ; Luis ROBLES-DIAZ ; Jose M GARCIA-BUENO ; Teresa RAMON Y CAJAL ; Elena HERNANDEZ-AGUDO ; Victoria HEREDIA SOTO ; Ivan MARQUEZ-RODAS ; Maria Jose ECHARRI ; Carmen LACAMBRA-CALVET ; Raquel SAEZ ; Maite CUSIDO ; Andres REDONDO ; Luis PAZ-ARES ; David HARDISSON ; Marta MENDIOLA ; Jose PALACIOS ; Javier BENITEZ ; Maria Jose GARCIA
Journal of Gynecologic Oncology 2016;27(1):e7-
OBJECTIVE: We aimed to evaluate the prognostic and predictive value of the nucleotide excision repair-related gene GTF2H5, which is localized at the 6q24.2-26 deletion previously reported by our group to predict longer survival of high-grade serous ovarian cancer patients. METHODS: In order to test if protein levels of GTF2H5 are associated with patients' outcome, we performed GTF2H5 immunohistochemical staining in 139 high-grade serous ovarian carcinomas included in tissue microarrays. Upon stratification of cases into high- and low-GTF2H5 staining categories (> and < or = median staining, respectively) Kaplan-Meier and log-rank test were used to estimate patients' survival and assess statistical differences. We also evaluated the association of GTF2H5 with survival at the transcriptional level by using the on-line Kaplan-Meier plotter tool, which includes gene expression and survival data of 855 high-grade serous ovarian cancer patients from 13 different datasets. Finally, we determined whether stable short hairpin RNA-mediated GTF2H5 downregulation modulates cisplatin sensitivity in the SKOV3 and COV504 cell lines by using cytotoxicity assays. RESULTS: Low expression of GTF2H5 was associated with longer 5-year survival of patients at the protein (hazard ratio [HR], 0.52; 95% CI, 0.29 to 0.93; p=0.024) and transcriptional level (HR, 0.80; 95% CI, 0.65 to 0.97; p=0.023) in high-grade serous ovarian cancer patients. We confirmed the association with 5-year overall survival (HR, 0.55; 95% CI, 0.38 to 0.78; p=0.0007) and also found an association with progression-free survival (HR, 0.72; 95% CI, 0.54 to 0.96; p=0.026) in a homogenous group of 388 high-stage (stages III-IV using the International Federation of Gynecology and Obstetrics staging system), optimally debulked high-grade serous ovarian cancer patients. GTF2H5-silencing induced a decrease of the half maximal inhibitory concentration upon cisplatin treatment in GTF2H5-silenced ovarian cancer cells. CONCLUSION: Low levels of GTF2H5 are associated with enhanced prognosis in high-grade serous ovarian cancer patients and may contribute to cisplatin sensitization.
Adult
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Aged
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Aged, 80 and over
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Biomarkers, Tumor/biosynthesis/genetics
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Cystadenocarcinoma, Serous/*genetics/metabolism/pathology
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Female
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Gene Expression Regulation, Neoplastic
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Humans
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Kaplan-Meier Estimate
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Middle Aged
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Neoplasm Grading
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Neoplasm Proteins/biosynthesis/genetics
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Neoplasms, Glandular and Epithelial/*genetics/metabolism/pathology
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Ovarian Neoplasms/*genetics/metabolism/pathology
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Prognosis
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Transcription Factors/biosynthesis/*genetics
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Tumor Cells, Cultured