Paracoccidioidomycosis is the most important systemic mycosis in Latin America. The main entrance of the fungus is the airway. It primarily occurs in the lung, but in its disseminated form may affect any organ. The liver is one of the organs afflicted by this disease and its homeostasis may be impaired. The aim of the present study was to evaluate the evolution of paracoccidioidomycosis in the liver of Swiss mice and correlate morphological factors with the expression of gp43 and with physicochemical analysis via FT-Raman of the infected organ. According to colony forming unit (CFU) and granuloma counting, the first and second weeks were the periods when infection was most severe. Tissue response was characterized by the development of organized granulomas and widespread infection, with yeasts located within the macrophages and isolated hepatocytes. The gp43 molecule was distributed throughout the hepatic parenchyma, and immunostaining was constant in all observed periods. The main physicochemical changes of the infected liver were observed in the spectral ranges between 1700–1530 cm−1 and 1370 – 1290 cm−1, a peak shifting center attributed to phenylalanine and area variation of -CH2 and -CH3 compounds associated to collagen, respectively. Over time, there was a direct proportional relationship between the number of CFUs, the number of granulomas and the physicochemical changes in the liver of mice infected with Paracoccidioides brasiliensis. The expression of gp43 was similar in all observed periods.
Paracoccidioides brasiliensis (Pb) can disseminate through the lymphatic and hematogenic pathways. As a result the spleen and other lymphoid organs are targets of paracoccidioidomycosis. There are few studies describing this disease in the spleen and more detailed descriptions are required. This study combines Histopathology and Fourier Transform Raman Spectroscopy (FT‐Raman) methods to study spleen infected by Pb. The Swiss mice were euthanized after 1, 2, 4 and 8 weeks of infection with Pb, and their spleens were removed for ex vivo analyzes. Histopathological evaluation revealed that the red pulp was the area most affected, presenting the highest concentration of yeasts, inflammatory cells and cells in apoptosis, with no observation of typical granuloma formation. The main physical–chemical changes were detected in the spectral ranges between 1730–1590 cm−1 and 1390–1280 cm−1 attributed to amide I and vibrational deformation mode of CH3 and CH2 molecules, respectively. The amide I was correlated to total protein content and CH3 and CH2 molecules to collagen of the spleen. The association between histological and physical–chemical methods enabled detection of several alterations in the spleen, including apoptosis, contributing to a better understanding of paracoccidioidomycosis, a tropical neglected disease.
In this work, the application of photoacoustic and photothermal methods for the evaluation of physicochemical properties of biological systems will be presented as well as the state of art of these methods fostering clinical diagnostics.
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