Inflammatory-Targeted Lipid Carrier as a New Nanomaterial to Formulate an Inhaled Drug Delivery System.

There is a pressing need for efficacious therapies in the field of respiratory diseases and infections. Lipid nanocarriers, administered through aerosols, represent a promising tool for maximizing therapeutic concentration in targeted cells and minimizing systemic exposure. However, this approach requires the application of efficient and safe nanomaterials. Palmitoylethanolamide (PEA), an endocannabinoid-like endogenous lipid, plays a crucial role in providing protective mechanisms during inflammation, making it an interesting material for preparing inhalable lipid nanoparticles (LNPs). This report aims to preliminarily explore the in vitro behavior of LNPs prepared with PEA (PEA-LNPs), a new inhalable inflammatory-targeted nanoparticulate drug carrier. PEA-LNPs exhibited a size of about 250 nm, a rounded shape, and an marked improvement in PEA solubility in comparison to naked PEA, indicative of easily disassembled nanoparticles. A twin glass impinger instrument was used to screen the aerosol performance of PEA-LNP powders, obtained via freeze-drying in the presence of two quantities of mannose as a cryoprotectant. Results indicated that a higher amount of mannose improved the emitted dose (ED), and in particular, the fine particle fraction (FPF). A cytotoxicity assay was performed and indicated that PEA-LNPs are not toxic towards the MH-S alveolar macrophage cell line up to concentrations of 0.64 mg/mL, and using coumarin-6 labelled particles, a rapid internalization into the macrophage was confirmed. This study demonstrates that PEA could represent a suitable material for preparing inhalable lipid nanocarrier-based dry powders, which signify a promising tool for the transport of drugs employed to treat respiratory diseases and infections.

Autonomic Regulation of Facial Temperature during Stress: A Cross-Mapping Analysis.

Skin temperature reflects the Autonomic Nervous System (ANS)'s response to emotions and mental states and can be remotely measured using InfraRed Thermography. Understanding the physiological mechanisms that affect facial temperature is essential to improve the precision of emotional inference from thermal imaging. To achieve this aim, we recorded thermal images from 30 volunteers, at rest and under acute stress induced by the Stroop test, together with two autonomic correlates, i.e., heart rate variability and electrodermal activity, the former serving as a measure of cardiovascular dynamics, and the latter of the activity of the sweat glands. We used a Cross Mapping (CM) approach to quantify the nonlinear coupling of the temperature from four facial regions with the ANS correlates. CM reveals that facial temperature has a statistically significant correlation with the two autonomic time series, under both conditions, which was not evident in the linear domain. In particular, compared to the other regions, the nose shows a significantly higher link to the electrodermal activity in both conditions, and to the heart rate variability under stress. Moreover, the cardiovascular activity seems to be primarily responsible for the well-known decrease in nose temperature, and its coupling with the thermal signals significantly varies with gender.

Towards a Contactless Stress Classification Using Thermal Imaging.

Thermal cameras capture the infrared radiation emitted from a body in a contactless manner and can provide an indirect estimation of the autonomic nervous system (ANS) dynamics through the regulation of the skin temperature. This study investigates the contribution given by thermal imaging for an effective automatic stress detection with the perspective of a contactless stress recognition system. To this aim, we recorded both ANS correlates (cardiac, electrodermal, and respiratory activity) and thermal images from 25 volunteers under acute stress induced by the Stroop test. We conducted a statistical analysis on the features extracted from each signal, and we implemented subject-independent classifications based on the support vector machine model with an embedded recursive feature elimination algorithm. Particularly, we trained three classifiers using different feature sets: the full set of features, only those derived from the peripheral autonomic correlates, and only those derived from the thermal images. Classification accuracy and feature selection results confirmed the relevant contribution provided by the thermal features in the acute stress detection task. Indeed, a combination of ANS correlates and thermal features achieved 97.37% of accuracy. Moreover, using only thermal features we could still successfully detect stress with an accuracy of 86.84% in a contact-free manner.

Discriminating Stress From Cognitive Load Using Contactless Thermal Imaging Devices.

This study proposes long wave infrared technology as a contactless alternative to wearable devices for stress detection. To this aim, we studied the change in facial thermal distribution of 17 healthy subjects in response to different stressors (Stroop Test, Mental Arithmetic Test). During the experimental sessions the electrodermal activity (EDA) and the facial thermal response were simultaneously recorded from each subject. It is well known from the literature that EDA can be considered a reliable marker for the psychological state variation, therefore we used it as a reference signal to validate the thermal results. Statistical analysis was performed to evaluate significant differences in the thermal features between stress and non-stress conditions, as well as between stress and cognitive load. Our results are in line with the outcomes of previous studies and show significant differences in the temperature trends over time between stress and resting conditions. As a new result, we found that the mean temperature changes of some less studied facial regions, e.g., the right cheek, are able not only to significantly discriminate between resting and stressful conditions, but also allow to recognize the typology of stressors. This outcome not only directs future studies to consider the thermal patterns of less explored facial regions as possible correlates of mental states, but more importantly it suggests that different psychological states could potentially be discriminated in a contactless manner.

Quantitative analysis of histidine decarboxylase gene (hdcA) transcription and histamine production by Streptococcus thermophilus PRI60 under conditions relevant to cheese making.

This study evaluated the influence of parameters relevant for cheese making on histamine formation by Streptococcus thermophilus. Strains possessing a histidine decarboxylase (hdcA) gene represented 6% of the dairy isolates screened. The most histaminogenic, S. thermophilus PRI60, exhibited in skim milk a high basal level of expression of hdcA, upregulation in the presence of free histidine and salt, and repression after thermization. HdcA activity persisted in cell extracts, indicating that histamine might accumulate after cell lysis in cheese.

Identification of a tyrosine decarboxylase gene (tdcA) in Streptococcus thermophilus 1TT45 and analysis of its expression and tyramine production in milk.

In this study, a tyrosine decarboxylase gene (tdcA) was identified in 1 among 83 Streptococcus thermophilus strains tested. Its sequence, nearly identical to that of a tdcA of Lactobacillus curvatus, indicated a horizontal gene transfer event. Transcription in milk and the formation of critical levels of tyramine were observed in the presence of tyrosine.

Application of AFLP fingerprint analysis for studying the biodiversity of Streptococcus thermophilus.

Streptococcus thermophilus is a lactic acid bacteria (LAB) widely used in milk fermentation processes as a starter culture. In this work the genetic diversity of S. thermophilus isolates from different sources was analyzed using Amplified Fragment Length Polymorphism fingerprinting (AFLP). Since this is the first report that indicates the application of AFLP in order to study genotypic polymorphism in S. thermophilus species, an optimization of experimental conditions was carried out to decide the optimal AFLP analysis protocol. Furthermore the fingerprinting resolutions of AFLP and RAPD (Random Amplified Polymorphic DNA) were evaluated and compared. The overall data suggest that genotypic characterization performed by AFLP provide a better view of microbial diversity of S. thermophilus, indicating that RAPD is less discriminating than AFLP. The successful use of AFLP analysis in the characterization of S. thermophilus strains reported in this study suggests the potential uses for this technique to define the whole-genome diversity of each specific strain, as an alternative to the fingerprinting methods used till now.

Characterization of tetracycline-resistant Streptococcus thermophilus isolates from Italian soft cheeses.

Tetracycline-resistant Streptococcus thermophilus isolates from soft cheeses harbored the genes tet(S), tet(M), and tet(L). Molecular analysis of these genes revealed their expression, localization on plasmids or Tn916-Tn1545 family transposons, and their similarity with published sequences. The study highlights the importance of an accurate safety assessment of using S. thermophilus as a starter culture.

Molecular diversity and transferability of the tetracycline resistance gene tet(M), carried on Tn916-1545 family transposons, in enterococci from a total food chain.

In the present study, 20 enterococci belonging to the species Enterococcus faecalis (12 strains), Enterococcus faecium (4), Enterococcus durans (2), Enterococcus hirae (1) and Enterococcus mundtii (1) and originating from a total production chain of swine meat commodities were analysed to investigate the diversity of their tetracycline resistance gene tet(M). PCR-RFLP and sequence analysis showed that the tet(M) gene of most strains can be correlated with the Tn916 transposon. Conversely, tet(M) of six E. faecalis and the E. hirae strain, all isolated from pig faecal samples, may be associated with previously undescribed members of the Tn916-1545 transposon family. In vitro filter conjugation trials showed the ability of 50% of the enterococcal strains, including E. mundtii, to transfer the tet(M) gene (and the associated Tn916 and new transposons) to E. faecalis or Listeria innocua recipient strains. tet(M) gene transfer to L. innocua recipient was also directly observed in meat food products. Collectively, these sequence and conjugation data indicate that various transposons can be responsible of the spread of tetracycline resistance in enterococci and validate the opinion that Enterococcus species are important sources of antibiotic resistance genes for potentially pathogenic bacteria occurring in the food chain.