Gene expression, purification, and functional characterization of recombinant conotoxin μ-TIIIA and TIIIAlaMut in Escherichia coli with clinical evaluation of antiwrinkle efficacy

Several clinical studies on laser resurfacing on the neck have yielded variable results with adverse sequelae of hypopigmentation and scarring using the CO2 laser. The Er:YAG laser on the neck resulted in an improved appearance with no adverse sequelae. The combined CO2/Er:YAG laser is a laser that combines a low fluence CO2 laser with the Er:YAG laser in a near simultaneous beam. To study the effects of the CO2/Er:YAG laser on the neck. The decreased nonspecific thermal damage of the CO2/Er:YAG laser should result in decreased postoperative erythema and improved wound healing. Eleven patients were treated with the CO2/Er:YAG laser on the neck as well as the face at identical settings. The face was treated with four passes while the neck was treated with two passes. The patients were evaluated for the first 2 weeks then 3-6 months postoperatively. Patients were asked about treatment satisfaction and improvement in skin texture and color using a 25% scale. Skin texture and color, as well as improvement in wrinkling and adverse results were judged by a dermatologist using a 0-4 scale. Moderate improvement was seen in skin color while a higher degree of improvement was seen in skin texture and wrinkling in all 11 patients. No adverse sequelae, including hypopigmentation or scarring, were observed. The majority of patients had a 75-100% improvement in skin texture and color in addition to a 75-100% overall satisfaction rating. There is a higher degree of overall patient satisfaction, as well as improvement in skin texture and skin color, compared to patients treated with the Er:YAG laser alone. The CO2/Er:YAG laser is a highly effective laser for neck resurfacing with no adverse sequelae to date.

Structural Modeling of Toxin Interactions with the Human Voltage-Gated Sodium Channel Pore

SummaryVoltage-gated sodium (NaV) channels, initially characterized in excitable cells, have been shown to be aberrantly expressed in non-excitable cancer tissues and cells from epithelial origins such as in breast, lung, prostate, colon, and cervix, whereas they are not expressed in cognate non-cancer tissues. Their activity was demonstrated to promote aggressive and invasive potencies of cancer cells, both in vitro and in vivo, whereas their deregulated expression in cancer tissues has been associated with metastatic progression and cancer-related death. This review proposes NaV channels as pharmacological targets for anticancer treatments providing opportunities for repurposing existing NaV-inhibitors or developing new pharmacological and nutritional interventions.

In response to the growing consumer demand for highly effective anti-aging products, Science4Beauty LLC obtained a recombinant conotoxin μ-TIIIA in E. coli strain and developed a novel cosmetic formulation. The main active ingredient, recombinant conotoxin μ-TIIIA, has the ability to block Nav 1.4 voltage-gated sodium channels, thereby reducing muscle contractions and the appearance of dynamic wrinkles. Company comprehensive approach aims to address multiple aspects of skin ageing, offering a product – the MIORELAXANT MAGIC cream, not only provides immediate improvements in skin texture and wrinkle appearance but also significantly prolongs the action of botulinum toxin. The 60 volunteers suffering from facial wrinkles around the lateral corners of the eyes, randomly divided into two groups – patients who underwent treatment with botulinum toxin in the area of the circular muscles of the eyes, they used the tested cosmetic products (Study Cream and Placebo) for 6 months after the treatment. The conducted research indicates a clinically significant use of MIORELAXANT MAGIC primarily in enhancing/extending the effects of aesthetic treatments using botulinum toxin and reducing skin seborrhea. Moreover, this product shows strong, positive and statistically significant effect on improving hydration, elasticity and skin tone, which justifies its use in a wide group of the population.

Protein-Protein Interactions Based Drug Discovery Against the Voltage-Gated Sodium Channel

The aim of this study was to evaluate the efficacy and safety of injectable bio-revitalizers and rejuvenative therapies licensed under Medicines and Healthcare products Regulatory Agency (MHRA) and National Institute for Health and Care Excellence (NICE) guidelines. These included platelet-rich plasma (PRP) and exosome-based treatments. This review synthesizes clinical outcomes, safety profiles and patient-reported satisfaction to provide evidence-based recommendations for advancing dermatological practice in the UK. A systematic review was conducted following the PRISMA guidelines. Databases including PubMed, MEDLINE, Embase and Cochrane Library were searched for studies published between 2010 and 2023. Keywords included ‘injectable bio-revitalisers’, ‘hyaluronic acid’, ‘poly­nucleotides’, ‘platelet-rich plasma’, ‘exosome therapy’, ‘efficacy’ and ‘safety’. Inclusion criteria focused on licensed products approved in the UK under MHRA and NICE regulations, such as Profhilo®, Restylane® Skinboosters, Sunekos®, Nucleofill®, Juvelook®, PRP and exosome-based therapies. Data extraction covered clinical efficacy (e.g. hydration, elasticity, wrinkle reduction), safety (e.g. adverse events, tolerability) and patient-reported outcomes. Twenty-six studies involving 2450 patients were included. Key findings highlighted consistent efficacy and safety across therapies. Hyaluronic acid-based bio-revitalizers (e.g. Profhilo®, Restylane® Skinboosters) improved hydration (20–25%), elasticity (15–18%) and wrinkle reduction, with patient satisfaction rates of 88–90%. Adverse events were limited to transient erythema and swelling. Polynucleotide-based therapies (e.g. Nucleofill®, Juvelook®) enhanced skin regeneration, provided antioxidant benefits and showed sustained results up to 6 months, with minimal adverse effects and improved firmness (18–22%). PRP showed moderate-to-significant improvements in skin texture and fine lines, with 76% of patients reporting enhanced skin quality. Adverse events were mild. Exosome-based therapies had emerging evidence indicating improvements in tone, texture and collagen stimulation, with high satisfaction rates (89%) and minimal adverse events. In conclusion, injectable bio-revitalizers, including PRP and exosome-based therapies, demonstrate high efficacy and safety profiles when adhering to MHRA and NICE guidelines. These therapies provide innovative, minimally invasive options for skin rejuvenation, with consistent patient satisfaction. Standardized treatment protocols, tailored patient selection criteria, and long-term studies are needed to optimize outcomes. PRP and exosome-based therapies expand regenerative dermatology and enhance patient care. Recommendations for practice are as follows: (i) incorporate PRP and exosome therapies into bio-revitalization strategies, emphasizing regenerative potential; (ii) develop national guidelines for standardized protocols; (iii) establish multicentre registries for tracking long-term outcomes and (iv) implement training programmes on appropriate use and safety considerations. This review offers a comprehensive evaluation of licensed bio-revitalizers and rejuvenate therapies, providing colleagues with evidence-based insights for improving patient care and advancing practice in aesthetic and therapeutic dermatology. The findings serve as a valuable teaching tool, fostering reflection on current practices and encouraging innovation within the field.

Vitamins C, E, and A and substances of plant origin, including azelaic acid and phytic acid are frequently used in cosmetic preparations to counteract oxidative stress and negative effects of free radicals. The aim of the study was to evaluate a novel combined therapy consisting of azelaic acid, ascorbic acid, and phytic acid applied layer on layer. Twenty study participants received a series of eight treatments performed every 7 days. Twenty percent azelaic acid and then 30% phytic acid were applied to the entire face, while 40% l-ascorbic acid only on the left side. The preparations were applied layer by layer. Skin parameters were measured before the series of treatments (T0), after the series of eight treatments (T1-8 weeks), and 1 month after the end of the treatment (T2-12 weeks). The application of two and three active compounds resulted in a significant improvement in erythema and hyperpigmentation both on the forehead and the cheeks, however, more pronounced effects were observed when all the three active compounds were used. Both applied types of treatment considerably increased skin moisture. All the participants (100%) were satisfied with the effects of the treatment. A majority of them reported an improvement in skin hydration, firmness, and elasticity, more uniform skin tone and a reduction of skin redness and wrinkles. Topical application of these active compounds resulted in improvement of skin elasticity and flexibility, reduction of wrinkles, hyperpigmentation, erythema, and telangiectasia as well as amelioration of skin tone.

Neurotoxins are of interest as lead molecules for the development of pharmaceuticals and insecticides. Huwentoxin-IV (HwTx-IV) is a 35-residue peptide isolated from the venom of Chinese bird spider Cyriopagopus schmidti. It has six cysteine residues that form three disulfide bonds, which form a stable inhibitor cystine knot (ICK) motif. This neurotoxin is one of the most potent inhibitors of the human voltage-gated sodium channel (hNaV1.7) described to date. Humans have nine NaV channel subtypes with different roles and tissue distribution. NaV1.1–1.3 are found primarily in the central nervous system (CNS), NaV1.6 is found in both the CNS and nodes of Ranvier in the peripheral nervous system (PNS), NaV1.4 and NaV1.5 are found in skeletal and cardiac muscles, respectively, and NaV1.7–1.9 are found primarily in peripheral sensory neurons where they are involved in pain sensing. NaV1.7 plays a key role in the propagation of pain signals, and it is a validated analgesic target. HwTx-IV potently inhibits hNaV1.7 with an IC50 of ~20 nM but it inhibits the off-target cardiac subtype NaV1.5 with far lower potency.In this study, a rationally designed triple-mutant HwTx-IV analogue (E1G,E4G,Y33W) was produced recombinantly via expression in the periplasm of Escherichia coli. The mutant peptide (m3-HwTx-IV) has significantly increased potency against hNaV1.7 (IC50 = 0.4 ± 0.1 nM) without increased potency against hNaV1.5. Neither its activity against other subtypes nor its structure has been characterised. Bacterial expression enabled production of uniformly 15N/13C-labelled recombinant m3-HwTx-IV for atomic-resolution structure determination using multidimensional heteronuclear NMR spectroscopy. Additionally, the activity of the recombinant peptide on various NaV channel subtypes was measured via automated patch-clamp electrophysiology using human (HEK293 and CHO) cell lines stably expressing the NaV channels. In parallel the analgesic efficacy of the NaV1.7 potency-optimised peptide was investigated using a rodent pain model.m3-HwTx-IV and similar spider-venom peptides from the same family inhibit NaV channel activation by binding to the domain II voltage sensor (VSDII) of the channel. In this project, I applied NMR-based methods to probe the atomic details of the interaction between gating-modifier venom peptides and the isolated VSDII of several human NaV subtypes, including hNaV1.7, hNaV1.6, and hNaV1.1. Key channel-interacting residues were identified by titration of the isolated VSDs with isotope-labelled gating-modifier peptides using solution NMR. The knowledge gained from this work, in combination with other studies such as the recently elucidated cryo-EM structures of NaV1.7, provides a platform for structure-guided rational engineering of subtype-selective inhibitors of human NaV channels as potential therapeutics for the treatment of chronic pain.

Molecular determinants for the subtype specificity of μ-conotoxin SIIIA targeting neuronal voltage-gated sodium channels

Tarantula Toxin SGTx-1 alters Gating Kinetics of Human Voltage-Gated Sodium Channel Nav1.7

Unique electrophysiological property of a novel Nav1.7, Nav1.8, and Nav1.9 sodium channel blocker, ANP-230

The voltage-gated sodium (NaV) channel subtype NaV1.7 plays a critical role in pain signaling, making it an important drug target. Here we studied the molecular interactions between μ-Conotoxin KIIIA (KIIIA) and the human NaV1.7 channel (hNaV1.7). We developed a structural model of hNaV1.7 using Rosetta computational modeling and performed in silico docking of KIIIA using RosettaDock to predict residues forming specific pairwise contacts between KIIIA and hNaV1.7. We experimentally validated these contacts using mutant cycle analysis. Comparison between our KIIIA-hNaV1.7 model and the cryo-EM structure of KIIIA-hNaV1.2 revealed key similarities and differences between NaV channel subtypes with potential implications for the molecular mechanism of toxin block. The accuracy of our integrative approach, combining structural data with computational modeling, experimental validation, and molecular dynamics simulations, suggests that Rosetta structural predictions will be useful for rational design of novel biologics targeting specific NaV channels.

There are several treatment modalities for mild to moderate photodamage. The demand for effective treatments with minimal side effects has increased. The objective of this study was to determine the effectiveness of short-pulsed erbium:YAG laser in treating mild to moderate photodamage. Twenty patients were treated with the short-pulse erbium:YAG laser on the face and neck. One pass was given over the entire face with two to three passes over the perioral and periorbital regions. Patients were evaluated for improvement of pigmentary irregularities, skin texture, and fine wrinkles. All side effects were recorded at follow-up visits. Two lasers were used, Sciton and Cynosure CO3, under local anesthesia. There was a 58% reduction in pigment irregularities and a 54% improvement in skin texture. There was minimal improvement in fine wrinkles with one pass. Two and three passes resulted in a 50% reduction in wrinkles. The procedure was minimally painful. Side effects included 3 to 5 days of erythema and edema. Patients returned to work within 3 days on average. There were no infections. Patient satisfaction with the procedure was rated as very good. There was no difference in efficacy or adverse effects with either laser. One-pass short-pulse erbium:YAG laser in treating photodamage of the face and neck is safe and effective. There are minimal side effects and patients heal within 3 to 5 days.

Introduction: Young and healthy skin is a beauty standard that generates the necessity of acceptance. Collagen supplements have recently grown in popularity and become a trendy product advertised as a remedy for various skin problems. As it turns out, numerous scientific studies have shown the benefits of hydrolyzed collagen supplementation in improving the signs of skin ageing. There are various animal sources of collagen, but marine fish collagen has the best safety profile, biocompatibility and bioavailability and remains the most frequent product used in scientific trials. Among newest randomised-control trials five were researched for this study to evaluate the efficacy of collagen supplementation and its influence of skin appearance with both objective and subjective assessment. State of knowledge: hydrolysed collagen supplementation demonstrated a significant reduction in wrinkles, improvement in skin texture, firmness and appearance. Moreover, studies show significant enhancement of the water content in the stratum corneum and epidermis. Reduction of transepidermal water loss and increased level of natural moisturising factor was observed. Conclusions: hydrolysed collagen supplements can delay and improve the signs of skin ageing by decreasing facial wrinkles and improving skin hydration and elasticity. Supplement intake is effective and safe - analysed studies have not reported any major adverse effects. Since reviewed trials lasted for no longer than 12 weeks, further studies are needed to evaluate the long-term use of hydrolysed collagen peptides.

Background/Objectives: This study aimed to develop and evaluate an anti-pollution film-forming spray (FFS) containing coffee cherry pulp extract (FFS-CCS). The formulation was designed to create a protective skin barrier, improving skin health while defending against environmental pollutants. Its physical properties, dust resistance, stability, skin penetration, and clinical effectiveness were assessed to ensure optimal performance and safety. Methods: Various polymers and a ternary solvent system were used to enhance the stability and solubility of bioactive compounds from the coffee cherry pulp extract. The formulations were characterized based on appearance, film formation, viscosity, pH, spray uniformity, spray pattern, angle, film thickness, and particle adhesion. Stability testing was conducted under different storage conditions. Skin penetration was assessed using Franz diffusion cells with Strat-M® membranes to simulate human skin. A single-blind, placebo-controlled trial with 42 participants was conducted over 60 days to evaluate the effects of FFS-CCS on skin hydration, tone, and wrinkle reduction. Clinical assessments were performed using a Corneometer, Mexameter, and Skin Visioscan. Results: The FFS1-CCS formulation, incorporating PVP K90 and a ternary solvent system, significantly improved the solubility, stability, and bioavailability of key bioactive compounds (chlorogenic acid, caffeine, and theophylline). Physical characterization confirmed uniform, transparent films with optimal viscosity and sprayability. Stability testing showed minimal degradation. Skin penetration and retention studies revealed enhanced retention of bioactive compounds with minimal systemic absorption. PVP K90, along with ethanol and propylene glycol, extended the compounds' residence time on the skin, ensuring localized delivery. Clinically, FFS1-CCS significantly improved skin hydration, reduced roughness, lightened skin tone, and decreased erythema. Conclusions: The FFS1-CCS formulation utilizing PVP K90 significantly enhanced the stability, bioavailability, and skin retention of coffee cherry pulp extract, resulting in improved skin hydration, wrinkle reduction, and skin tone enhancement. These findings highlight the potential of coffee cherry pulp extract as a multifunctional, sustainable cosmeceutical ingredient, offering both anti-aging and environmental protection benefits, making it a promising solution for skincare applications.