Evaluating biochemical differences in thyroglobulin from normal and goiter tissues by infrared spectral imaging.

Thyroglobulin is a glycoiodoprotein that is produced by thyroid follicular cells; it is stored in follicles in structures known as colloids. The main function of this protein is to stock the hormones triiodothyronine (T3) and thyroxine (T4) until the body requires them. This study aims to demonstrate that infrared spectral imaging with appropriate multivariate analysis can reveal biochemical changes in this glycoprotein. The results achieved herein point out biochemical differences in the colloid samples obtained from normal and goiter patients including glycosylation and changes in the secondary conformational structure. We have presented the first spectral histopathology-based method to detect biochemical differences in thyroid colloids, such as TG iodination, glycosylation, and changes in the secondary structure in normal and goiter patients. The observed changes in the colloids were mainly due to the alterations in amide I and amide II (secondary conformation of proteins) and there is a correlation with different glycosylation between normal and goiter tissues.

Deep neural networks can differentiate thyroid pathologies on infrared hyperspectral images.

BACKGROUND AND OBJECTIVE: The thyroid is a gland responsible for producing important body hormones. Several pathologies can affect this gland, such as thyroiditis, hypothyroidism, and thyroid cancer. The visual histological analysis of thyroid specimens is a valuable process that enables pathologists to detect diseases with high efficiency, providing the patient with a better prognosis. Existing computer vision systems developed to aid in the analysis of histological samples have limitations in distinguishing pathologies with similar characteristics or samples containing multiple diseases. To overcome this challenge, hyperspectral images are being studied to represent biological samples based on their molecular interaction with light. METHODS: In this study, we address the acquisition of infrared absorbance spectra from each voxel of histological specimens. This data is then used for the development of a multiclass fully-connected neural network model that discriminates spectral patterns, enabling the classification of voxels as healthy, cancerous, or goiter. RESULTS: Through experiments using the k-fold cross-validation protocol, we obtained an average accuracy of 93.66 %, a sensitivity of 93.47 %, and a specificity of 96.93 %. Our results demonstrate the feasibility of using infrared hyperspectral imaging to characterize healthy tissue and thyroid pathologies using absorbance measurements. The proposed deep learning model has the potential to improve diagnostic efficiency and enhance patient outcomes.

Cell death mechanisms in Leishmania amazonensis triggered by methylene blue-mediated antiparasitic photodynamic therapy.

Antiparasitic photodynamic therapy (ApPDT) is an emerging approach to manage cutaneous leishmaniasis (CL) since no side effects, contraindications and parasite resistance have been reported. In addition, methylene blue (MB) is a suitable photosensitizer to mediate ApPDT on CL. In this study we aimed to look for the best parameters to eradicate Leishmania amazonensis and investigated the cell death pathways involved in MB-mediated ApPDT. MB uptake by parasites was determined using different MB concentrations (50, 100, 250 and 500 µM) and incubation times (10, 30 and 60 min). L. amazonensis promastigotes were cultured and submitted to ApPDT using different concentrations of MB (50, 100 and 250 µM) combined to a red LED emitting at 645 ±â€¯10 nm. The pre-irradiation time was 10 min. Two optical powers (100 mW and 250 mW) were tested and cells were exposed to 60 and 300 s of MB-mediated ApPDT delivering energies of 6, 15, 30 and 75 J and fluences of 21.2, 53.1, 106.2 and 265.4 J/cm2, respectively. Following ApPDT, cells were prepared for flow cytometry and transmission electron microscopy to unravel the mechanisms of cell death. Our results showed the lowest MB concentration (50 µM) and the lowest optical power (100 mW) promoted the highest percentage of cell decrease. ApPDT caused alterations on cell membrane permeability as well depolarization of mitochondrial membrane potential. We also observed ultrastructural changes of the parasites such as cell shrinkage, intense vacuolization of the cytoplasm, enlargement of mitochondrion-kinetoplast complex, and small blebs on parasite flagella and cell membrane after MB-mediated ApPDT. Taken together, our findings ratify that ApPDT parameters play a pivotal role in cell susceptibility and suggest that apoptosis is involved in parasite death regardless MB-mediated ApPDT protocol.

Optical diagnosis of actinic cheilitis by infrared spectroscopy.

Actinic cheilitis (AC) is considered a potentially malignant disorder of the lip. Biomolecular markers study is important to understand malignant transformation into squamous cell carcinoma. Fourier transform infra red (FT-IR) spectroscopy was used to analyze AC in this study. OBJECTIVES: The aim of the study was to evaluate if FT-IR spectral regions of nucleic acids and collagen can help in early diagnosis of malignant transformation. METHODS: Tissues biopsies of 14 patients diagnosed with AC and 14 normal tissues were obtained. FT-IR spectra were measured at five different points resulting in 70 spectra of each. Analysis of Principal components analysis (PCA) and linear discrimination analysis (LDA) model were also used. In order to verify the statistical difference in the spectra, Mann-Whitney U test was performed in each variable (wavenumber) with p-value <0.05. RESULTS: After the Mann-Whitney U test the vibrational modes of CO (Collagen 1), PO2 (Nucleic Acids) and CO asymmetric (Triglycerides/Lipids) were observed as a possible spectral biomarker. These bands were chosen because they represent the vibrational modes related to collagen and DNA, which are supposed to be changed in AC samples. Based on the PCA-LDA results, the predictive model corresponding to the area under the curve was 0.91 for the fingerprint region and 0.83 for the high wavenumber region, showing the greater accuracy of the test. CONCLUSIONS: FT-IR changes in collagen and nucleic acids could be used as molecular biomarkers for malignant transformation.

The chemokines secretion and the oxidative stress are targets of low-level laser therapy in allergic lung inflammation.

Recent studies show that low-level laser therapy (LLLT) has an important anti-inflammatory action in acute lung inflammation. The present work explored if laser therapy is able to antagonize eosinophils and allergic inflammation induced by oxidative stress in Balb/c mice. Forty-eight hours after challenge, the leukocyte counting, ROS and nitrite/nitrate level, RANTES, CCL3, CCL8 as well as eotaxins were measured in the bronchoalveolar lavage fluid (BALF) of laser-treated mice or not. Into the lung, some chemokines receptors, the iNOS activity and mRNA expression, and the activities of superoxide dismutase (SOD), catalase, gluthatione, NADPH oxidase activities and thiobarbituric acid reactive species (T-Bars) were measured. Laser-treated allergic mice presented reduction of both the ICAM-1 and eosinophil in the lungs. RANTES, CCL8, CCL3 and eotaxins were reduced in BALF of laser-treated allergic mice. In allergic mice lung LLLT decreased the CCR1 and CCR3 and restored the oxidative stress balance as well. Laser decreased the lipidic peroxidation in allergic mice lung as much as increased SOD, GPx and GR. It shows that LLLT on allergic lung inflammation involves leukocyte-attractant chemokines and endogenous antioxidant. Based on results, LLLT may ultimately become a non- invasive option in allergic lung disease treatment. The top figure illustrates the laser decreasing the eosinophils migration into BALF and the bottom figure shows the laser upregulating the expression of heme-oxygenase (anti-oxidant enzyme) in lung tissue anti-oxidant.

Cell death mechanisms in Leishmania amazonensis triggered by methylene blue-mediated antiparasitic photodynamic therapy

Antiparasitic photodynamic therapy (ApPDT) is an emerging approach to manage cutaneous leishmaniasis (CL) since no side effects, contraindications and parasite resistance have been reported. In addition, methylene blue (MB) is a suitable photosensitizer to mediate ApPDT on CL. In this study we aimed to look for the best parameters to eradicate Leishmania amazonensis and investigated the cell death pathways involved in MB-mediated ApPDT. MB uptake by parasites was determined using different MB concentrations (50, 100, 250 and 500 µM) and incubation times (10, 30 and 60 min). L. amazonensis promastigotes were cultured and submitted to ApPDT using different concentrations of MB (50, 100 and 250 µM) combined to a red LED emitting at 645 ±â€¯10 nm. The pre-irradiation time was 10 min. Two optical powers (100 mW and 250 mW) were tested and cells were exposed to 60 and 300 s of MB-mediated ApPDT delivering energies of 6, 15, 30 and 75 J and fluences of 21.2, 53.1, 106.2 and 265.4 J/cm2, respectively. Following ApPDT, cells were prepared for flow cytometry and transmission electron microscopy to unravel the mechanisms of cell death. Our results showed the lowest MB concentration (50 µM) and the lowest optical power (100 mW) promoted the highest percentage of cell decrease. ApPDT caused alterations on cell membrane permeability as well depolarization of mitochondrial membrane potential. We also observed ultrastructural changes of the parasites such as cell shrinkage, intense vacuolization of the cytoplasm, enlargement of mitochondrion-kinetoplast complex, and small blebs on parasite flagella and cell membrane after MB-mediated ApPDT. Taken together, our findings ratify that ApPDT parameters play a pivotal role in cell susceptibility and suggest that apoptosis is involved in parasite death regardless MB-mediated ApPDT protocol

A thermal investigation of dental bleaching in vitro.

OBJECTIVE: Our goal was to investigate the surface temperature variations in the cervical region via infrared thermography, as well as the temperature within the pulp chamber via thermocouples, of mandibular incisors when subjected to dental bleaching using two different 35% hydrogen peroxide gels, red (HP) and green (HPM), when activated by halogen light (HL) and LED light. BACKGROUND DATA: Temperatures increases of more than 5.5 degrees C are considered to be potentially threatening to pulp vitality, while those higher than 10 degrees C can result in periodontal injury. MATERIALS AND METHODS: Tooth samples were randomly divided into four groups (n = 10 each), according to the bleaching agent and catalyst light source used. RESULTS: Mean values and standard deviations of the temperature increases inside the pulp chamber in the HL groups were 4.4 degrees +/- 2.1 degrees C with HP, and 4.5 degrees +/- 1.2 degrees C with HPM; whereas in the groups using LED light, they were 1.4 degrees +/- 0.3 degrees C for HP, and 1.5 degrees +/- 0.2 degrees C for HPM. For the root surfaces, the maximum temperature increases in the groups irradiated with HL were 6.5 degrees +/- 1.5 degrees C for HP, and 7.5 degrees +/- 1.1 degrees C with HPM; whereas in the groups irradiated with LED light, they were 2.8 degrees +/- 0.7 degrees C with HP, and 3 degrees +/- 0.8 degrees C with HPM. There were no statistically significant differences in pulp and surface temperature increases between the groups using different gels, although the mean temperature increases were significantly higher for the groups irradiated with HL when compared with those irradiated with the LED light (p < 0.05 with Tukey's test). CONCLUSION: LED light may be safe for periodontal and pulp tissue when using this method, but HL should be used with care.