Browsing by Subject "RT-PCR"

Now showing 1 - 5 of 5
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    Butylated hydroxytoluene induces type-V collagen and overexpression of remodeling genes/proteins in experimental lung fibrosis
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Martins, Vanessa; Teodoro, Walcy Rosolia; Pereira Velosa, Ana Paula; Andrade, Priscila; Farhat, Cecília; Fabro, Alexandre Todorovic; Capelozzi, Vera Luiza
    Anomalous histoarchitecture with increased levels of type-V collagen (Col V) in lungs of human idiopathic pulmonary fibrosis (IPF) and bleomycin (BLM) airway-centered interstitial fibrosis suggest that this collagen can be a possible trigger involved in the pathogenesis of these diseases. Butylated hydroxytoluene (BHT) injury model revealed a distal involvement of lung parenchyma with significant endothelial injury and fibrotic response, contrasting with the BLM airway-centered insult. We undertook this study to analyze whether BHT alters distal airway/alveolar epithelial cells (AECs) and extracellular matrix (ECM) signaling involved in the initiation and progression of pulmonary fibrosis in a different pathway concerning overexpression of Col V. Female mice C57BL/6 (n=6) were instilled intraperitoneally with 400mg/kg of BHT dissolved in 1 mL of corn oil and euthanized at day 14 or 21 after BHT administration. Morphometry, immunohistochemistry and transmission electron microscopy were performed to characterize microscopic and submicroscopic changes of AECs and endothelial cells through transforming growth factor beta (TGF-β) basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) expression. Immunofluorescence and immunogold electron microscopy were performed to characterize Col V. Quantitative polymerase chain reaction (qPCR) was used to confirm differential levels of RNA messenger. BHT lungs showed marked fibrotic areas and hyperplastic AECs. The alveolar damage caused destruction of elastic fibers and a critical increase of Col V in ECM of distal lung parenchyma. Fibrogenesis-promoting markers TGF-β, bFGF and VEGF were also overexpressed in situ, coinciding with up-regulation in remodeling enzymes, growth factors, cytokines, transduction and transcription genes. BHT alters distal lung parenchyma signaling involved in pulmonary fibrosis highlighted similarities to human IPF in a pathway involving Col V arising as a promissory model to identify effective therapeutic targets.
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    Confirmation of SARS-CoV-2 airborne dissemination indoors using "COVID-19 traps"
    (Elsevier Ltd., 2021-12-22) Navas-Carrillo, Diana; Moreno-Docón, Antonio; Ortega-García, Juan A.; Torres-Cantero, Alberto M.; Garcia-Vázquez, Elisa; Ramírez, Pablo; Orenes-Piñero, Esteban; Cirugía, Pediatría y Obstetricia y Ginecología
    Understanding the capacity of SARS-CoV-2 virus to infect by surfaces and by airborne dissemination has become in one of the most important issues in the world nowadays. Thus, the aim of this study was to confirm aerosol dissemination from patients with coronavirus infection using “COVID-19 traps” that included different untouched surfaces within them. In a previous study, we evaluated the presence of the virus and its stability in 6 different surfaces placed in the rooms of 6 patients with a positive diagnostic of COVID-19. For that, we performed a case series study with 180 samples collected from the surfaces included into the “COVID-19 traps” located in the rooms of patients in a COVID-19 ward unit (CWU) at a Spanish referral hospital at 24, 48 and 72 h. RNA was extracted from the surfaces with a swab and subsequently analyzed with RT-PCR to evaluate the presence of the virus and its stability. Positives were found in almost all the rooms and at all analyzed times. Surfaces could not be touched by patients or health workers, so viral spreading was unequivocally produced by airborne. In addition, ROC curves were performed to corroborate that airborne dissemination was directly associated with patients’ viral load. Thus, aerosol dissemination and patients’ viral load were confirmed as key parameters for SARS-CoV-2 infection. Our results point the importance of SARS-CoV-2 virus airborne dissemination indoors and may shed some light in this debate.
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    Confirmation of SARS-CoV-2 airborne dissemination indoors using “COVID-19 traps”
    (Elsevier, 2021-12-15) Navas-Carrillo, Diana; Moreno-Docón, Antonio; Ortega-García, Juan A.; Torres-Cantero, Alberto M.; García-Vázquez, Elisa; Ramírez, Pablo; Orenes-Piñero, Esteban; Medicina
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    Expressions of CXCL12, CXCL10 and CCL18 in Warthin tumors characterized pathologically by having a lymphoid stroma with germinal centers
    (Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2021) Mochizuki, Kunio; Oishi, Naoki; Masataka Kawai, Masataka Kawai; Odate, Toru; Tahara, Ippei; Kasai, Kazunari; Kondo, Tetsuo
    The Warthin tumor is a benign neoplasm of the salivary glands, histologically, the tumor has an oncocytic epithelial component forming uniform rows of cells surrounded by cystic spaces associated with a lymphoid stroma often showing the presence of germinal centers. The lymphoid stroma is a representative microscopic finding. If this lymphocytic accumulation is active, some sort of transmitter should exist between the Warthin tumor cells and lymphocytes. C-X-C motif chemokine ligand (CXCL) 12, CXCL10 and C-C motif chemokine ligand 18 (CCL18) are a chemoattractant for lymphocytes in vivo. There is no report on the relationship between these chemokines and Warthin tumors. In this study, we investigated these chemokines expressions in 20 Warthin tumors using immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). For comparison, we also enrolled samples of pleomorphic adenoma, which is another benign salivary gland tumor type without prominent lymphocytic infiltration. All Warthin tumors were immunopositive for CXCL12 and CXCL10, and these reactivities were diffuse. Meanwhile, the majority of pleomorphic adenomas were immunonegative for CXCL12 (95%), CXCL10 (80%) and CCL18 (85%). Warthin tumor and pleomorphic adenoma cases were significantly different in these immunostaining expressions (CXCL12, p<0.001; CXCL10, p<0.001; CCL18, p=0.024). We examined CXCL12, CXCL10 and CCL18 mRNA expressions of 3 representative Warthin tumor samples, each having these chemokines immunopositive areas detected by RT-PCR. Finding CXCL12 and CXCL10 expressions indicate that these chemokines may play a part in the formation of a lymphoid stroma within Warthin tumors. In regards to this phenomenon, the participation of CCL18 might be restrictive compared to CXCL12 and CXCL10.
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    vidences of SARS-CoV-2 virus air transmission indoors using several untouched surfaces: A pilot study
    (Elsevier, 2021-01-10) Baño, Francisco; Navas, Diana; Moreno-Docon, Antonio; Marin, juana María; Misiego, Rocio; Ramirez, Pablo; Orenes-Piñero, Esteban; Cirugía, Pediatría y Obstetricia y Ginecología
    Nowadays, there is an important controversy about coronavirus air transmission. The aim of this study was to determine aerosol transmission from patients with coronavirus infection using "COVID-19 traps" that included different untouched surfaces within them. 42 swab samples of 6 different surfaces placed in the rooms of 6 patients with a positive diagnostic of COVID-19 were analyzed with RT-PCR technique to evaluate the presence of the virus and its stability. Samples were collected at 24, 48 and 72 h. Patients were in an intensive care unit (ICU) and in a COVID-19 ward unit (CWU) at a Spanish referral hospital. None of the samples placed in the ICU unit were positive for COVID-19. However, two surfaces, placed in a CWU room with a patient that required the use of respiratory assistance were positive for coronavirus at 72 h. Surfaces could not be touched by patients or health workers, so viral spreading was unequivocally produced by air transmission. Thus, fomites should be considered as a possible mode of transmission of coronavirus and frequent disinfection of surfaces should be taken into account. Our results, although preliminary, point the importance of SARS-CoV-2 virus air transmission indoors and may shed some light in this debate.