Proliferation and Invasion of Melanoma Are Suppressed by a Plant Protease Inhibitor,Leading to Downregulation of Survival/Death-Related Proteins

Cell adhesion and migration are crucial for cancer progression and malignancy. Drugs available for the treatment of metastatic melanoma are expensive and unfit for certain patients. Therefore, there is still a need to identify new drugs that block tumor cell development. We investigated the effects of Enterolobium contortisiliquum trypsin inhibitor (EcTI), a protease inhibitor, on cell viability, cell migration, invasion, cell adhesion, and cell death (hallmarks of cancer) in vitro using human melanoma cells (SK-MEL-28 and CHL-1). Although EcTI did not affect non-tumor cells, it significantly inhibited the proliferation, migration, invasion, and adhesion of melanoma cells. Investigation of the underlying mechanisms revealed that EcTI triggered apoptosis and nuclear shrinkage, increased PI uptake, activated effector caspases-3/7, and produced reactive oxygen species (ROS). Furthermore, EcTI disrupted the mitochondrial membrane potential, altered calcium homeostasis, and modified proteins associated with survival and apoptosis/autophagy regulation. Acridine orange staining indicated acidic vesicular organelle formation upon EcTI treatment, demonstrating a cell death display. Electronic microscopy corroborated the apoptotic pattern by allowing the visualization of apoptotic bodies, mitochondrial cristae disorganization, and autophagic vesicles. Taken together, these results provide new insights into the anti-cancer properties of the natural EcTI protein, establishing it as a promising new therapeutic drug for use in melanoma treatment.     

Effects of Deacetylasperulosidic Acid on Atopic Dermatitis through Modulating Immune Balance and Skin Barrier Function in HaCaT,HMC-1, and EOL-1 Cells

The medicinal plant noni (Morinda citrifolia) is widely dispersed throughout Southeast Asia, the Caribbean, and Australia. We previously reported that fermented Noni could alleviate atopic dermatitis (AD) by recovering Th1/Th2 immune balance and enhancing skin barrier function induced by 2,4-dinitrochlorobenzene. Noni has a high deacetylasperulosidic acid (DAA) content, whose concentration further increased in fermented noni as an iridoid constituent. This study aimed to determine the anti-AD effects and mechanisms of DAA on HaCaT, HMC-1, and EOL-1 cells. DAA inhibited the gene expression and secretion of AD-related cytokines and chemokines including interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-25, IL-33, thymic stromal lymphopoietin, tumor necrosis factor-alpha, monocyte chemoattractant protein-1, thymus and activation-regulated chemokine, macrophage-derived chemokine, and regulated upon activation, normal T cell expressed and secreted, in all cells, and inhibited histamine release in HMC-1 cells. DAA controlled mitogen-activated protein kinase phosphorylation levels and the translocation of nuclear factor-kappa light chain enhancer of activated B cells into the nucleus by inhibiting IκBα decomposition in all the cells. Furthermore, DAA increased the expression of proteins involved in skin barrier functions such as filaggrin and involucrin in HaCaT cells. These results confirmed that DAA could relieve AD by controlling immune balance and recovering skin barrier function.     

Micellar electrokinetic chromatographic method for the dabrafenib determination in biological samples

Two different micellar electrokinetic chromatographic methods to determine dabrafenib in urine and serum, both using borate buffer (pH 9.2, 20 mM) and SDS as separation electrolyte, are developed and validated. The analyses were carried out in a fused‐silica capillary of 75 μm of internal diameter and total length of 47 and 37 cm for urine and serum determination, respectively. The detection of the target compound was performed at 227 nm in urine samples and at 251 nm in serum samples. The linearity range was from 1 to 21 mg/L of dabrafenib in urine and from 2 to 40 mg/L in serum. In all cases, inter‐ and intraday RSDs were <4%. Sample preparation of serum samples consists of an only step of 1:1 dilution with water before its injection in the electrophoretic system. These simple, sensitive, accurate, and cost‐effective methods can be used in routine clinical practice to monitor dabrafenib concentrations in urine and serum of metastatic melanoma skin cancer patients.     

Use of Phosphoantimonic Acid for Arsenic Determination by Activation Analysi

A methed was elaborated to determine arsenic amount in skin samples by neutron activation analysis and high-resolution gamma spectrometry including sample preparation after mineralization by sorption on silicagel saturated with phosphoantimonic acid. The samples were irradiated in the VVR-S reactor core in a flux of 10¹³ neutrons · cm^?2 · s^?1, mineralized with a mixture of nitric and sulphuric acids, the gamma spectrum was measured after passing through an ion-exchanger containing 0.5 mol P₂O₅ to 1 mol Sb₂O₅ (SP-2) by a gamma-spectroscopy system PLURIMAT 20 with a Ge(Li) detector, and the ⁷⁶As photopeak of E_γ = 0.559 MeV was measured. The values of the results obtained varied in the range of (4?5 )× 10^?1 ppm arsenic. The sensitivity of the proposed determination method is l×10^?3 μg arsenic.     

Design and Fabrication of Artificial Skin: Chitosan and Gelatin Immobilization on Silicone by Poly Acrylic Acid Graft Using a Plasma Surface Modification Method

Acrylic acid (AAc) was grafted onto the surfaces of polydimethylsiloxane (PDMS) films using a two-step oxygen plasma treatment. The first step of this method included oxygen plasma pretreatment of the PDMS films, immersion in AAc, and drying. The second step was carried out by plasma polymerization of the preadsorbed reactive AAc on the surfaces of the dried pretreated films. Then chitosan and gelatin were immobilized onto the poly(acrylic acid) grafted silicone through covalent bonding. The films were characterized by attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy and water contact angle measurements. Fibroblast cells (L929) were cultured onto the chitosan- and gelatin-immobilized poly(acrylic acid)-grafted silicone and poly(acrylic acid)-grafted silicone films. It was observed that the chitosan- and gelatin-immobilized surfaces showed significant cell growth in comparison with poly(acrylic acid)-grafted silicone samples. It seems that chitosan- and gelatin-immobilized surfaces may have an excellent potential to be used as a derm-like matrix.     

Development of a HPLC method for determination of four UV filters in sunscreen and its application to skin penetration studies

This study describes the development, validation and application of a high‐performance liquid chromatography (HPLC) method for the simultaneous determination of the in vitro skin penetration profile of four UV filters on porcine skin. Experiments were carried out on a gel‐cream formulation containing the following UV filters: diethylamino hydroxybenzoyl hexyl benzoate (DHHB), bis‐ ethylhexyloxyphenol methoxyphenyl triazine (BEMT), methylene bis‐ benzotriazolyl tetramethylbutylphenol (MBBT) and ethylhexyl triazone (EHT). The HPLC method demonstrated suitable selectivity, linearity (10.0–50.0 μg/mL), precision, accuracy and recovery from porcine skin and sunscreen formulation. The in vitro skin penetration profile was evaluated using Franz vertical diffusion cells for 24 h after application on porcine ear skin. None of the UV filters penetrated the porcine skin. Most of them stayed on the skin surface (>90%) and only BEMT, EHT and DHHB reached the dermis plus epidermis layer. These results are in agreement with previous results in the literature. Therefore, the analytical method was useful to evaluate the in vitro skin penetration of the UV filters and may help the development of safer and effective sunscreen products.     

Development and validation of a simple method for the extraction and quantification of crisaborole in skin layers

Crisaborole is a boron compound recently approved by the US Food and Drug Administration as a 2% ointment for the treatment of mild to moderate atopic dermatitis. This work describes a simple method for the quantification of the drug in the skin layers at the end of in‐vitro permeation experiments. Chromatographic separation was carried out on a reverse‐phase C₁₈ column using a mixture of trifluoroacetic acid 0.05%–acetonitrile (55:45, v/v) as mobile phase, pumped at 1 ml/min. Column temperature was 35°C and UV detection was performed at 250 nm. The method was linear in the range of concentration from 0.06 to 6 μg/ml (R² = 1) and was selective, precise and accurate. Depending on the solvent used, the LOQ ranged from 0.014 to 0.030 μg/ml and the LOD from 0.005 to 0.010 μg/ml. The extraction from all the skin layers was quantitative. The developed method was successfully tested in an in‐vitro permeation study, proving to be an effective tool in the development of new formulations containing crisaborole.     

Electrolytic Cell Design to Simulate the Electrochemical Skin Response

Small fiber peripheral neuropathy is an early complication of diabetes. Electric skin response to some stimulus, as electrochemical skin conductance ECS, is a promising route in the early follow‐up of such diseases. It is related to sweat gland innervations and their permeability to chlorides and protons; it is non‐invasive, quantitative and reproducible. In routine clinical use, it could allow to better adapt the treatments and improve the adhesion for preventing pathological progress, thus reducing colossal healthcare costs. To optimize the measurements and understand the electrochemical behavior of electrodes, an original electrolytic cell was designed in lab scale. Thereby, an electrolyte is chosen to mimic sweat composition. For achieving currents range of ESC in vivo measurements, the original idea was to play on electrolyte viscosity by adding sucrose. In this paper, the novel electrolytic lab cell is presented with its limiting kinetics processes. A model of chloride migration to the anode and global electric model dedicated to the cell are proposed. Cell parameters are thoroughly studied, e. g. the resistance, which is equivalent to the inverse of ESC, by exploiting the models and through in vitro experiments, with protocols focusing on reproducibility. This original approach establishes, inter alia, an important result: the resistance is accurately retrieved using linear voltammetry, whereas single voltage measurement fails notably and is, therefore, unsatisfactory.     

三聚化非厄密晶格中具有趋肤效应的拓扑边缘态

近年来,探索新的拓扑量子结构、深入分析各种多聚化拓扑晶格中的新奇物理性质已经成为热点.并且,多聚化拓扑模型在量子光学等领域的研究也愈发深入,拥有广阔的发展前景.本文聚焦于研究三聚化非厄密晶格中的新奇拓扑特性.首先,若晶胞内最近邻正反向耦合不相等,三聚化模型中的体态和边缘态出现趋肤效应.其中,随着最近邻耦合正反系数差的增大,拓扑保护的边缘态的宽度和简并度均可被调制,边缘态数量也会减少.其次,当在考虑次近邻耦合的影响时,随着次近邻耦合系数在适当范围内变化,系统本征能谱的上下能隙及其中具有趋肤效应的边缘态也会发生不对称的变化.此外,当适当改变两种耦合系数,三聚化非厄密模型的体态和边缘态的局域程度也会随之发生变化.