Trypanosoma cruzi is the etiological agent of Chagas disease, which is endemic in Latin America. Ursolic acid (UA) is a natural pentacyclic triterpene which has been shown to reduce the peak of parasitemia in T. cruzi infected mice. Due to UA was described as an inducer of autophagy and having into account that our previous work established the protective role of this process on in vivo infections, we decided to study the possible involvement of UA in the elimination of parasites in macrophages and cardiac cells.

To test this, we infected cells with T. cruzi for 24 hours, and then treated the samples for 24, 48 or 72 hours under both control and UA (10 µM) conditions, and evaluated the amount of amastigotes. Both xenophagy (by IFI) and ROS generation (by 2',7'-dichlorodihydrofluorescein diacetate reaction) were also tested as two possible mechanisms of action of this drug. Our data showed that UA decreased the amount of amastigotes in cells. We also observed that UA induces the autophagy pathway, and that LC3, the marker of autophagy, is recruited around amastigotes, indicating xenophagy of these parasites. Moreover, the production of ROS after 24 hours of treatment is increased on infected cells but, interestingly, UA does not have this effect on non-infected cells.

We conclude that this natural compound promotes parasite death through induction of autophagy and other host cell responses.

Discrimination of Acne Vulgaris with Human Scalp Hair tissues using - FTIR-ATR Spectroscopy

Padmavathi R^1*, Rajamannan B² , Gunasekaran S³, Ramkumar GR⁴, Sankari G⁵_, Muthu S⁶

1 Department of Physics, Meenakshi Sundararajan Engineering College, Kodambakkam, Chennai, 600024,TN, India.

2 Engineering physics, FEAT Annamalai University, Annamalai Nagar, 608002, Chidambaram, TN, India.

3 Research and development St. Peter’s institute of Higher Education and Research, St. Peter’s University, Avadi,

Chennai, 600054, TN, India.

4 Department of Physics, C. Kandaswaminaidu College for Men, Chennai-600102, India.

5 Department of Physics, Meenakshi College for Women’s, Kodambakkam, Chennai-600024, India.

6 Department of Physics, Govt. Thirumagal Mills College, Gudiyatham-632602, Vellore, TN, India.

Correspondence; pathmavati@gmail.com

Abstract: Acne vulgaris is a chronic skin disease, which occurs due to inflammation of the hair follicles and sebum-producing (sebaceous) glands of the skin called pilosebaceous unit and the anaerobic propionic acne bacterium, P. Acne. Human sebum is dominantly made up about 57.5% of triglycerides and fatty acids, 26%wax esters, 12% Squalene, and 4.5% Cholesterol. The increased level Androgen hormone, sebum lipid composition, P. Acne overgrowth which induces monocytes and pro-inflammatory cytokines attract neutrophils, basophils, and T cells to the pilosebaceous unit and drive epithelial hyperproliferation i.e., Acne vulgaris. The actual Biomolecular changes due to acne vulgaris disease are present in the blood, in the sebum, and in the noninvasive sample of human scalp hair follicles. The main objectives of the present study are to analyze human scalp hair follicles samples using FTIR-ATR spectroscopy to compare and discriminate the spectral signatures of acne vulgaris and healthy scalp hair samples through acne biomarkers Protein, Amide I, Amide II and Squalene (LDL), using the method of internal ratio parameters. This work represents a first step in the development of the analytical tool for future drug development.

Keywords: Acne Vulgaris, Hair tissue samples, Discrimination, FTIR-ATR.

The increased resistance of bacteria against conventional pharmaceutical solutions, the antibiotics, has raised serious health concerns. This has stimulated interest in the development of bio-based therapeutics with limited resistance, namely, essential oils (EOs) or antimicrobial peptides (AMPs). This study envisaged the evaluation of the antimicrobial efficacy of selected biomolecules, namely LL37, pexiganan, tea tree oil (TTO), cinnamon leaf oil (CLO) and niaouli oil (NO), against four bacteria commonly associated to nosocomial infections: Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Pseudomonas aeruginosa. The antibiotic vancomycin and silver nanoparticles (AgNPs) were used as control compounds for comparison purposes. The biomolecules were initially screened for their antibacterial efficacy using the agar-diffusion test, followed by the determination of minimal inhibitory concentrations (MICs), kill-time kinetics and the evaluation of the cell morphology upon 24 h exposure. All agents were effective against the selected bacteria. Interestingly, the AgNPs required a higher concentration (4000–1250 µg/mL) to induce the same effects as the AMPs (500–7.8 µg/mL). Pexiganan was the most effective biomolecule, requiring lower concentrations to kill both Gram-positive and Gram-negative bacteria (62.5-7.8 µg/mL), within a short period of time (averaging 2 h 15 min for all bacteria). Most biomolecules apparently disrupted the bacteria membrane stability due to the observed cell morphology deformation and by effecting on the intracellular space. AMPs were observed to induce morphological deformations and cellular content release, while EOs were seen to split and completely envelope bacteria. Data unraveled more of the potential of these new biomolecules as replacements for the conventional antibiotics and allowed us to take a step forward in the understanding of their mechanisms of action against infection-related bacteria.

Dengue virus is a serious public health issue in tropical and subtropical regions. The global incidence of dengue necessitates the potent antiviral medication for the prevention of proliferation of the virus inside the human body. The DEN2 NS2B/NS3 protease presents in the dengue virus, is an attractive drug target due to its essential role in viral replication, survival, and other cellular activities. In traditional medicine, Carica papaya leaves have been used for the treatment of dengue fever in Sri Lanka, Pakistan, Malaysia. Therefore, phytochemicals present in Carica papaya leaves, have a potential anti-viral activity and can be used as strong drug candidates against the dengue virus. In this investigation, two phytochemical compounds in Carica papaya leaves: 5,7-Dimethoxycoumarin and p-Coumaric acid were selected from the literature and then docked against the DEN2 NS2B/NS3 protease. The compounds showed strong interactions with favorable binding energies in the active site of DEN2 NS2B/NS3 protease. To validate the molecular docking results, the docked ligand-protein complexes were subjected to molecular dynamics simulation along with the apo form of the protein for 30 ns. The molecular dynamics simulation analysis comprising of root mean square deviation and fluctuation, the radius of gyration, hydrogen bonding, DSSP, and MM/PBSA revealed the stability of the apo and complex systems. Interactions formed by these compounds with residues Leu149 and Asn152 are found to be essential for the stability of the ligand-protein complex. The findings revealed that these phytochemical compounds depict the promising results against the DEN2 serotype of the dengue virus and the potential to work as therapeutic drugs. Further experimentation on the proposed compounds is necessary to validate the results and can lead to the development of strong inhibitors with improved activity.

Background: Pro-inflammatory cytokines and adipokines upregulation through NF-κB activation in adipose tissue has been considered to have an important function in the pathogenesis of obesity-related hypertension. Objective: to ascertain the effect of NF-κB inhibitor, (SN50) on TNF-α and AGT secretion and expression in mediating the anti-inflammatory effect through its effects on NF-κB activity in human adipose tissue. Methods: Primary human adipocytes were isolated from 10 overweight and 10 obese with and without hypertension, cultured, then treated with 10 ng/ml LPS, with and without NF-κB inhibitor, SN50 (50 μg/ml) at different time points. TNF-α secretion and NF-κB p65 activity were detected in supernatants extracted from cultured cell treated and untreated with LPS and SN50 by ELISA. NF-κB p65, TNF-α and AGT proteins expression were detected by western blot. TNF-α and AGT gene expression was detected in cells and performed using quantitative RT-PCR. The study was carried out at the Obesity Research Center, King Saud University, Riyadh, KSA. Results: Treatment of AbdSc adipocytes with LPS caused a significant increase in NF-κB p65 in overweight and obese, while, SN50-NF-κB inhibitor causes a reduction of NF-κB p65 in overweight and obese persons at all time points. Treatment of AbdSc adipocytes with LPS caused a significant increase in TNF-α secretion in overweight and obese subjects at all time points, whereas, SN50 leads to a decrease in TNF-α secretion at 3 and 12 hours. Treatment of with LPS caused increased TNF-α and AGT gene expression, whereas, in the presence of SN50, it reduces mRNA AGT levels. Conclusion: these adipokines with NF-κB activation may represent important biomarkers to evaluate hypertension risk and provide a mechanistic insight into the pathogenesis of obesity-related hypertension.