Academic Journal
An improved high-performance liquid chromatography (HPLC) method for detection of variations in the hydroxyproline content of tissue homogenates from Paracoccidioides brasiliensis-infected mice
| Title: | An improved high-performance liquid chromatography (HPLC) method for detection of variations in the hydroxyproline content of tissue homogenates from Paracoccidioides brasiliensis-infected mice |
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| Authors: | Magnus Ake Gidlund, Raphael Fagnani Sanchez Molina, Eva Burger |
| Source: | Journal of Venomous Animals and Toxins including Tropical Diseases, Vol 30 (2024) |
| Publisher Information: | SciELO, 2024. |
| Publication Year: | 2024 |
| Collection: | LCC:Arctic medicine. Tropical medicine LCC:Toxicology. Poisons LCC:Zoology |
| Subject Terms: | Paracoccidioidomycosis, Granulomatous lesions, Hydroxyproline, Collagen., Arctic medicine. Tropical medicine, RC955-962, Toxicology. Poisons, RA1190-1270, Zoology, QL1-991 |
| More Details: | Abstract Background: Paracoccidioidomycosis (PCM) is a severe granulomatous disease. The hallmark of this mycosis is fibrin degradation and granuloma formation as a result of a wound-healing process in the context of excessive inflammation. Therefore, as the content of collagen can be assessed by the methodology described in this manuscript, we propose that the content of hydroxyproline (HYP) be employed as a new and efficient measurement of granulomatous lesions developed. To estimate the level of HYP the major byproduct of the degradation process, we hypothesized that this simple and efficient technique could serve as a marker of disease severity. Methods: Five B10.A female mice were infected with P. brasiliensis and, after 15 days, the omentum was removed, subjected to histopathological analysis or processed (i.e. deproteinized and derivatized), and further analyzed on a reverse phase HPLC using a C-18 column. The omentum of five uninfected controls was also collected and similarly analyzed. Results: Infected mice showed numerous, disseminated paracoccidioidomycotic lesions, as well as marked collagen deposits, as observed in histopathologic analysis, and high levels of HYP. Normal uninfected mice showed no granulomas, little or no deposits of collagen fibers, and very low levels of HYP, as evaluated by HPLC. Our results show that the disease intensity as evaluated number and the morphology of the granulomatous lesions were correlated to the HYP levels using small tissue samples from the omentum, the main target organ of P. brasiliensis. Conclusions: Here we propose an alternative methodology to follow disease evolution and, to some extent, fungal load in experimental P. brasiliensis infection and suggest its usefulness to other diseases with pronounced fibrin degradation. |
| Document Type: | article |
| File Description: | electronic resource |
| Language: | English |
| ISSN: | 1678-9199 |
| Relation: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992024000100309&lng=en&tlng=en; http://www.scielo.br/pdf/jvatitd/v30/1678-9199-jvatitd-30-e20230068.pdf; https://doaj.org/toc/1678-9199 |
| DOI: | 10.1590/1678-9199-jvatitd-2023-0068 |
| Access URL: | https://doaj.org/article/2331b9fa2cea4dd0b6e4b5bd7d72926c |
| Accession Number: | edsdoj.2331b9fa2cea4dd0b6e4b5bd7d72926c |
| Database: | Directory of Open Access Journals |
| ISSN: | 16789199 |
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| DOI: | 10.1590/1678-9199-jvatitd-2023-0068 |
| Published in: | Journal of Venomous Animals and Toxins including Tropical Diseases |
| Language: | English |