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19 results on '"Pires, Layla"'

7. Ex vivo treatment of cytomegalovirus in human donor lungs using a novel chemokine-based immunotoxin

Author

Ribeiro, Rafaela V.P., Ku, Terrance, Wang, Aizhou, Pires, Layla, Ferreira, Victor H., Michaelsen, Vinicius, Ali, Aadil, Galasso, Marcos, Moshkelgosha, Sajad, Gazzalle, Anajara, Jeppesen, Mads G., Rosenkilde, Mette M., Liu, Mingyao, Singer, Lianne G., Kumar, Deepali, Keshavjee, Shaf, Sinclair, John, Kledal, Thomas N., Humar, Atul, Cypel, Marcelo, Ribeiro, Rafaela V.P., Ku, Terrance, Wang, Aizhou, Pires, Layla, Ferreira, Victor H., Michaelsen, Vinicius, Ali, Aadil, Galasso, Marcos, Moshkelgosha, Sajad, Gazzalle, Anajara, Jeppesen, Mads G., Rosenkilde, Mette M., Liu, Mingyao, Singer, Lianne G., Kumar, Deepali, Keshavjee, Shaf, Sinclair, John, Kledal, Thomas N., Humar, Atul, and Cypel, Marcelo

Abstract

Background: Transmission of latent human cytomegalovirus (HCMV) via organ transplantation with post-transplant viral reactivation is extremely prevalent and results in substantial adverse impact on outcomes. Therapies targeting the latent reservoir within the allograft to mitigate viral transmission would represent a major advance. Here, we delivered an immunotoxin (F49A-FTP) that targets and kills latent HCMV aiming at reducing the HCMV reservoir from donor lungs using ex-vivo lung perfusion (EVLP). Methods: HCMV seropositive human lungs were placed on EVLP alone or EVLP + 1mg/L of F49A-FTP for 6 hours (n = 6, each). CD14+ monocytes isolated from biopsies pre and post EVLP underwent HCMV reactivation assay designed to evaluate viral reactivation capacity. Off-target effects of F49A-FTP were studied evaluating cell death markers of CD34+ and CD14+ cells using flow cytometry. Lung function on EVLP and inflammatory cytokine production were evaluated as safety endpoints. Results: We demonstrate that lungs treated ex-vivo with F49A-FTP had a significant reduction in HCMV reactivation compared to controls, suggesting successful targeting of latent virus (76% median reduction in F49A-FTP vs 15% increase in controls, p = 0.0087). Furthermore, there was comparable cell death rates of the targeted cells between both groups, suggesting no off-target effects. Ex-vivo lung function was stable over 6 hours and no differences in key inflammatory cytokines were observed demonstrating safety of this novel treatment. Conclusions: Ex-vivo F49A-FTP treatment of human lungs targets and kills latent HCMV, markedly attenuating HCMV reactivation. This approach demonstrates the first experiments targeting latent HCMV in a donor organ with promising results towards clinical translation.

Published
2022

14. Inactivating hepatitis C virus in donor lungs using light therapies during normothermic ex vivo lung perfusion

Author

Galasso, Marcos, primary, Feld, Jordan J., additional, Watanabe, Yui, additional, Pipkin, Mauricio, additional, Summers, Cara, additional, Ali, Aadil, additional, Qaqish, Robert, additional, Chen, Manyin, additional, Ribeiro, Rafaela V. P., additional, Ramadan, Khaled, additional, Pires, Layla, additional, Bagnato, Vanderlei S., additional, Kurachi, Cristina, additional, Cherepanov, Vera, additional, Moonen, Gray, additional, Gazzalle, Anajara, additional, Waddell, Thomas K., additional, Liu, Mingyao, additional, Keshavjee, Shaf, additional, Wilson, Brian C., additional, Humar, Atul, additional, and Cypel, Marcelo, additional

Published
2019
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18. Optical clearing of melanoma in vivo: characterization by diffuse reflectance spectroscopy and optical coherence tomography.

Author

Pires, Layla, Demidov, Valentin, Vitkin, I. Alex, Bagnato, Vanderlei, Kurachi, Cristina, and Wilson, Brian C.

Subjects
*SKIN cancer diagnosis, *MELANOMA diagnosis, *TISSUE mechanics, *OPTICAL properties of skin, *RADIOSCOPIC diagnosis
Abstract

Melanoma is the most aggressive type of skin cancer, with significant risk of fatality. Due to its pigmentation, light-based imaging and treatment techniques are limited to near the tumor surface, which is inadequate, for example, to evaluate the microvascular density that is associated with prognosis. White-light diffuse reflectance spectroscopy (DRS) and near-infrared optical coherence tomography (OCT) were used to evaluate the effect of a topically applied optical clearing agent (OCA) in melanoma in vivo and to image the microvascular network. DRS was performed using a contact fiber optic probe in the range from 450 to 650 nm. OCT imaging was performed using a swept-source system at 1310 nm. The OCT image data were processed using speckle variance and depth-encoded algorithms. Diffuse reflectance signals decreased with clearing, dropping by ∼90% after 45 min. OCT was able to image the microvasculature in the pigmented melanoma tissue with good spatial resolution up to a depth of ∼300 μm without the use of OCA; improved contrast resolution was achieved with optical clearing to a depth of ∼750 μm in tumor. These findings are relevant to potential clinical applications in melanoma, such as assessing prognosis and treatment responses. Optical clearing may also facilitate the use of light-based treatments such as photodynamic therapy. [ABSTRACT FROM AUTHOR]

Published
2016
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19. Photodynamic Therapy in Pythium insidiosum – An In Vitro Study of the Correlation of Sensitizer Localization and Cell Death.

Author

Pires, Layla, Bosco, Sandra de Moraes Gimenes, Baptista, Maurício S., and Kurachi, Cristina

Subjects
*PHOTODYNAMIC therapy, *PYTHIUM, *ANTIBIOTICS, *CELL death, *COMMUNICABLE diseases, *IN vitro studies, *PHOTOSENSITIZERS
Abstract

Pythiosis is an infectious disease caused by Pythium insidiosum, a fungus-like organism. Due to the lack of ergosterol on its cell membrane, antibiotic therapy is ineffective. The conventional treatment is surgery, but lesion recurrence is frequent, requiring several resections or limb amputation. Photodynamic therapy uses photo-activation of drugs and has the potential to be an attractive alternative option. The in vitro PDT response on the growing of Pythium insidiosum culture was investigated using three distinct photosensitizers: methylene blue, Photogem, and Photodithazine. The photosensitizer distribution in cell structures and the PDT response for incubation times of 30, 60, and 120 minutes were evaluated. Methylene blue did not penetrate in the pathogen's cell and consequently there was no PDT inactivation. Photogem showed heterogenous distribution in the hyphal structure with small concentration inside the cells. Porphyrin-PDT response was heterogenous, death and live cells were observed in the treated culture. After 48 hours, hyphae regrowth was observed. Photodithazine showed more homogenous distribution inside the cell and with the specific intracellular localization dependent on incubation time. Photodithazine first accumulates in intracellular vacuoles, and at incubation times of one hour, it is located at all cell membranes. Higher inhibition of the growing rates was achieved with Photodithazine -PDT, over 98%. Our results showed that the photosensitizers that cross more efficiently the Pythium insidiosum membranes are able to cause extensive damage to the organism under illumination and therefore, are the best options for clinical treatment. [ABSTRACT FROM AUTHOR]

Published
2014
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