We encapsulated cisplatin into stealth pH-sensitive liposomes and studied their stability, cytotoxicity and accumulation in a human small-cell lung carcinoma cell line (GLC4) and its resistant subline (GLC4/CDDP). Since reduced cellular drug accumulation has been shown to be the main mechanism responsible for resistance in the GLC4/CDDP subline, we evaluated the ability of this new delivery system to improve cellular uptake. The liposomes were composed of dioleoylphosphatidylethanolamine (DOPE), cholesteryl hemisuccinate (CHEMS), and distearoylphosphatidylethanolamine-polyethyleneglycol 2000 (DSPE-PEG 2000 ) and were characterized by determining the encapsulation percentage as a function of lipid concentration. Among the different formulations, DOPE/CHEMS/DSPE-PEG liposomes (lipid concentration equal to 40 mM) encapsulated cisplatin more efficiently than other concentrations of liposomes (about 20.0%, mean diameter of 174 nm). These liposomes presented good stability in mouse plasma which was obtained using a 0.24-M EDTA solution (70% cisplatin was retained inside the liposomes after 30 min of incubation). Concerning cytotoxic effects, they are more effective (1.34-fold) than free cisplatin for growth inhibition of the human lung cancer cell line A549. The study of cytotoxicity to GLC4 and GLC4/ CDDP cell lines showed similar IC 50 values (approximately 1.4 µM), i.e., cisplatin-resistant cells were sensitive to this cisplatin formulation. Platinum accumulation in both sensitive and resistant cell lines followed the same pattern, i.e., approximately the same intracellular platinum concentration (4.0 x 10 -17 mol/cell) yielded the same cytotoxic effect. These results indicate that long-circulating pH-sensitive liposomes, also termed as stealth pH-sensitive liposomes, may present a promising delivery system for cisplatin-based cancer treatment. This liposome system proved to be able to circumvent the cisplatin resistance, whereas it was not observed when using non-long-circulating liposomes composed of phosphatidylcholine, phosphatidylserine, and cholesterol.

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Abstract-The temporal component of videos provides an important clue for activity recognition, as a number of activities can be reliably recognized based on the motion information. In view of that, this work proposes a novel temporal stream for two-stream convolutional networks based on images computed from the optical flow magnitude and orientation, named Magnitude-Orientation Stream (MOS), to learn the motion in a better and richer manner. Our method applies simple nonlinear transformations on the vertical and horizontal components of the optical flow to generate input images for the temporal stream. Experimental results, carried on two well-known datasets (HMDB51 and UCF101), demonstrate that using our proposed temporal stream as input to existing neural network architectures can improve their performance for activity recognition. Results demonstrate that our temporal stream provides complementary information able to improve the classical two-stream methods, indicating the suitability of our approach to be used as a temporal video representation.

While a large number of surveillance cameras available nowadays provide a safe environment, the huge amount of data generated by them prevents a manual processing, requiring the application of automated methods to understand the scene. However, the majority of the currently available methods are still unable to process this amount of data in real time, mainly those focusing on pedestrian detection. To optimize pedestrian detection methods, this work proposes a novel approach that performs a random filtering supported by the Maximum Search Problem theorem to select a very small number from all possible detection windows. Although the random filtering is able to select regions that capture every person on an image, some windows can cover only parts of a person, diminishing the accuracy. To solve that, a regression is applied to adjust the windows to the person's location. The computational cost reduction comes from the fact that the proposed approach does not need to perform any processing while selecting windows, differently from cascades of rejection that must evaluate at least simple features for every window. The experiments performed using a pedestrian detection based on Partial Least Squares show that the approach is effective in both accuracy and computational cost reduction.

Brucella abortus is a Gram-negative bacterium responsible for a worldwide zoonotic infection—Brucellosis, which has been associated with high morbidity rate in humans and severe economic losses in infected livestock. The natural route of infection is through oral and nasal mucosa but the invasion process through host gut mucosa is yet to be understood. Studies have examined the role of NLRP6 (NOD-like receptor family pyrin domain-containing-6 protein) in gut homeostasis and defense against pathogens. Here, we investigated the impact of gut microbiota and NLRP6 in a murine model of Ba oral infection. Nlrp6-/- and wild-type (WT) mice were infected by oral gavage with Ba and tissues samples were collected at different time points. Our results suggest that Ba oral infection leads to significant alterations in gut microbiota. Moreover, Nlrp6-/- mice were more resistant to infection, with decreased CFU in the liver and reduction in gut permeability when compared to the control group. Fecal microbiota transplantation from WT and Nlrp6-/- into germ-free mice reflected the gut permeability phenotype from the donors. Additionally, depletion of gut microbiota by broad-spectrum-antibiotic treatment prevented Ba replication in WT while favoring bacterial growth in Nlrp6-/-. Finally, we observed higher eosinophils in the gut and leukocytes in the blood of infected Nlrp6-/- compared to WT-infected mice, which might be associated to the Nlrp6-/- resistance phenotype. Altogether, these results indicated that gut microbiota composition is the major factor involved in the initial stages of pathogen host replication and partially also by the resistance phenotype observed in Nlrp6 -/- mice regulating host inflammation against Ba infection.