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Contreras, M. A.

Publications and source records attributed to Contreras, M. A..

2 recordsLinked to original sources

Discrete Protein Metric (DPM): A new image similarity metric to calculate accuracy of deep learning-generated cell focal adhesion predictions

Understanding cell behaviors can provide new knowledge on the development of different pathologies. Focal adhesion (FA) sites are important sub-cellular structures that are involved in these processes. To better facilitate the study of FA sites, deep learning (DL) can be used to predict FA site morphology based on limited datasets (e.g., cell membrane images). However, calculating the accuracy score of these predictions can be challenging due to the discrete/point pattern like nature of FA sites. In the present work, a new image similarity metric, discrete protein metric (DPM), was developed to calculate FA prediction accuracy. This metric measures differences in distribution (d), shape/size (s), and angle (a) of FA sites between the predicted image and its ground truth image. Performance of the DPM was evaluated by comparing it to three other commonly used image similarity metrics: Pearson correlation coefficient (PCC), feature similarity index (FSIM), and Intersection over Union (IoU). A sensitivity analysis was performed by comparing changes in each metric value due to quantifiable changes in FA site location, number, aspect ratio, area, or orientation. Furthermore, accuracy score of DL-generated predictions was calculated using all four metrics to compare their ability to capture variation across samples. Results showed better sensitivity and range of variation for DPM compared to the other metrics tested. Most importantly, DPM had the ability to determine which FA predictions were quantitatively more accurate and consistent with qualitative assessments. The proposed DPM hence provides a method to validate DL-generated FA predictions and can be extended to evaluating other predicted or segmented discrete structures of biomedical relevance.

bioengineering↗

New insights on the interaction mechanism of rhTNF with its antagonists Adalimumab and Etanercept

TNF is a pro-inflammatory cytokine that is a therapeutic target for inflammatory autoimmune disorders. Thus, TNF antagonists are successfully used for the treatment of these disorders. Here, new association patterns of rhTNF and its antagonists Adalimumab and Etanercept are disclosed. Active rhTNF was purified by IMAC from the soluble fraction of transformed E. coli. Protein detection was assessed by SDS-PAGE and western blot. The KD values for rhTNF interactions with their antagonists were obtained by non-competitive ELISA and by microscale thermophoresis. Molecular sizes of the complexes were characterized by SEC-HPLC. Surprisingly, both antagonists recognized the monomeric form of rhTNF under reducing and non-reducing conditions, indicating unexpected bindings of the antagonists to lineal epitopes and to one protomer of rhTNF. Binding curves of two phases with low and high KD values (<10-9 M and >10-8 M) were observed during thermophoresis experiments, suggesting the generation of complexes with different stoichiometry, which were confirmed by SEC-HPLC. This pioneer investigation revealed interactions of rhTNF with Adalimumab and Etanercept never described before, which constitute valuable data for future approaches into the study of their interaction mechanism.

molecular biology↗