Search

Purpose: Our goal was to study whether glycerol and xylitol provide protection against osmotic stress in keratinocytes. Methods: The experiments were performed on HaCaT keratinocytes. Hyperosmotic stress was induced by the addition of sorbitol (450, 500 and 600 mOsm). Both polyols were applied at two different concentrations (glycerol: 0.027% and 0.27%, xylitol: 0.045% and 0.45%). Cellular viability and cytotoxicity were assessed, intracellular Ca2+ concentration was measured, and the RNA expression of inflammatory cytokines was determined by means of PCR. Differences among groups were analyzed with one-way ANOVA and Holm-Sidak post-hoc test. When the normality test failed, Kruskal-Wallis one-way analysis of variance on ranks, followed by Dunn's method for pairwise multiple comparison was performed. Results: The higher concentrations of the polyols were effective. Glycerol ameliorated the cellular viability while xylitol prevented the rapid Ca2+ signal. Both polyols suppressed the expression of IL-1α but only glycerol decreased the expression of IL-1β and NFAT5. Conclusions: Glycerol and xylitol protect keratinocytes against osmotic stress. Despite their similar chemical structure, the effect of these polyols displayed differences. Hence, joint application of glycerol and xylitol may be a useful therapeutic approach for different skin disorders.

Fractional calculus is an emerging topic in artificial neural network training, especially when using gradient-based methods. This paper brings the idea of fractional derivatives to spiking neural network training using Caputo derivative-based gradient calculation. We focus on conducting an extensive investigation of performance improvements via a case study of small-scale networks using derivative orders in the unit interval. With particle swarm optimization we provide an example of handling the derivative order as an optimizable hyperparameter to find viable values for it. Using multiple benchmark datasets we empirically show that there is no single generally optimal derivative order, rather this value is data-dependent. However, statistics show that a range of derivative orders can be determined where the Caputo derivative outperforms first-order gradient descent with high confidence. Improvements in convergence speed and training time are also examined and explained by the reformulation of the Caputo derivative-based training as an adaptive weight normalization technique.

Petra Hartmann,1 Edina Butt,2 Ágnes Fehér,1 Ágnes Lilla Szilágyi,1 Kurszán Dávid Jász,1 Boglárka Balázs,3 Mónika Bakonyi,3 Szilvia Berkó,3 Gábor Erős,4 Mihály Boros,1 Gyöngyi Horváth,5 Endre Varga,2 Erzsébet Csányi3 1Institute of Surgical Research, University of Szeged, Szeged, Hungary; 2Department of Traumatology, University of Szeged, Szeged, Hungary; 3Department of Pharmaceutical Technology, University of Szeged, Szeged, Hungary; 4Department of Pathology, University of Szeged, Szeged, Hungary; 5Department of Physiology, University of Szeged, Szeged, Hungary Purpose: Since electroporation (EP) can increase the permeability of biological membranes, we hypothesized that it offers an opportunity to enhance the transdermal delivery of drugs for intra-articular indications. Our aim was to compare the anti-inflammatory and analgesic efficacy of EP-combined topical administration of diclofenac sodium hydrogel (50 mg mL-1 in 230 μL volume) with that of an equivalent dose of oral (75 mg kg-1) and simple topical administration. Methods: Arthritis was induced with the injection of 2% λ-carrageenan and 4% kaolin into the right knee joints of male Sprague Dawley rats. EP was applied for 8 min with 900 V high-voltage pulses for 5 ms followed by a 20 ms break. Drug penetration into the synovial fluid and plasma was detected by high-performance liquid chromatography. Leukocyte–endothelial interactions were visualized by intravital videomicroscopy on the internal surface of the synovium. Inflammation-induced thermal and mechanical hyperalgesia reactions, knee joint edema, and inflammatory enzyme activities were assessed at 24 and 48 h after arthritis induction. Results: EP significantly increased the plasma level of diclofenac as compared with the topical controls 10 min after the 2% λ-carrageenan and 4% kaolin injection. Increased leukocyte–endothelial interactions were accompanied by joint inflammation, which was significantly reduced by oral and EP diclofenac (by 45% and by 30%, respectively) and only slightly ameliorated by simple topical diclofenac treatment (by 18%). The arthritis-related secondary hyperalgesic reactions were significantly ameliorated by oral and EP-enhanced topical diclofenac treatments. The knee cross-section area (which increased by 35%) was also reduced with both approaches. However, simple topical application did not influence the development of joint edema and secondary hyperalgesia. Conclusion: The study provides evidence for the first time of the potent anti-inflammatory and analgesic effects of EP-enhanced topical diclofenac during arthritis. The therapeutic benefit provided by EP is comparable with that of oral diclofenac; EP is a useful alternative to conventional routes of administration. Keywords: diclofenac, transdermal delivery, HPLC, intravital videomicroscopy

The aim of this work was the formulation and comparison of different drug carrier systems for the transdermal delivery of lidocaine. Quality target product profile and quality attributes were identified, and an initial risk assessment was made according to the Quality by Design methodology. The critical quality attributes influencing the quality and efficacy of the final drug formulation were then defined in order to select the control points. Four formulation types, each containing 5% lidocaine were compared: conventional hydrogel, oleogel, lyotropic liquid crystal and nanostructured lipid carrier. Microscopic analysis, particle size and zeta potential measurements, Fourier transform infrared spectroscopy and Raman spectroscopy were performed to characterize the critical parameters in case of the different vehicles. Membrane diffusion and penetration studies were completed as well for each formulation in vitro and ex vivo, followed by measurements on skin hydration and transepidermal water loss in vivo. Our results lead us to the conclusion that the nanostructured lipid carrier is the most promising vehicle for the topical delivery of lidocaine. It showed the best penetration properties through heat separated epidermis and the highest moisturizing effect which are the most critical parameters based on the Quality by Design initial risk assessment and evaluation.

As drug development is extremely expensive, the identification of novel indications for in-market drugs is financially attractive. Multiple algorithms are used to support such drug repurposing, but highly reliable methods combining simulation of intracellular networks and machine learning are currently not available. We developed an algorithm that simulates drug effects on the flow of information through protein-protein interaction networks, and used support vector machine to identify potentially effective drugs in our model disease, psoriasis. Using this method, we screened about 1,500 marketed and investigational substances, identified 51 drugs that were potentially effective, and selected three of them for experimental confirmation. All drugs inhibited tumor necrosis factor alpha-induced nuclear factor kappa B activity in vitro, suggesting they might be effective for treating psoriasis in humans. Additionally, these drugs significantly inhibited imiquimod-induced ear thickening and inflammation in the mouse model of the disease. All results suggest high prediction performance for the algorithm.

Noninvasive body shaping is becoming a growing demand. The aim of this study was to investigate the efficacy and safety of the combined treatments of 1064 nm Nd:YAG and 2940 nm Er:YAG in noninvasive lipolysis and skin tightening. Ten females were enrolled, and all women's side of the waist or the lower part of the abdomen were treated. In the first step, the 1064 nm Nd:YAG was used. As a second step, the 2940 nm Er:YAG laser was applied. Each woman was treated four times, once every 2 weeks. The effects were determined by comparative photo documentation, waist circumference measurement, two‐dimensional B‐mode ultrasonography and low‐dose native computer tomography (CT), whereas body fat was monitored with bioelectric impedance. The tissue firmness was measured by ultrasound shear wave elastography. Combined laser treatment significantly reduced waist circumference and total body fat. Ultrasonography has revealed that the treatment considerably decreased fat thickness and improved skin stiffness in the treated region. Subcutaneous fat volume, measured by low‐dose CT, displayed a moderate decrease in the waist region. The combined 1064 nm Nd:YAG and 2940 nm Er:YAG laser treatment results in the reduction of fat tissue and tightens the skin as confirmed by objective measurements., Noninvasive body shaping is a growing demand. Our goal was to test the efficacy of the combined treatments of 1064 nm Nd:YAG and 2940 nm Er:YAG in noninvasive lipolysis and skin tightening. Four treatments were performed in 10 subjects. Waist circumference, fat thickness, fat volume and tissue firmness were determined. The combined 1064 nm Nd:YAG and 2940 nm Er:YAG laser treatment resulted in the reduction of fat tissue and tightened the skin.

Aim of the Study: During the reconstruction of alar defects involving the upper lip, reconstructive surgeons face the need for various thicknesses of tissues crucial to preserving the facial sulcus which is important for a cosmetically acceptable result. Our aim was to reconstruct the deep perialar and thinner lateral nasal alar defect in a single step procedure with a suitable flap which is reliable, has appropriate blood supply and provides an esthetically good result. Materials and Methods: Extended alar defect was reconstructed with a combined flap in 10 cases. During the procedure, a subcutaneous pedicle was created and the proximal part of the flap was rotated into the defect as a rotational flap. The procedure and the follow-up have been photo-documented in all cases. Furthermore, the perfusion of the flaps was monitored by means of laser Doppler flowmetry. Postoperative complications were evaluated with a semi-quantitative score and the patients completed a patient satisfaction questionnaire, too. Results: An optimal esthetic result was obtained in all cases after the operation. The lateral nasal alar part of the defect was reconstructed with the thinner proximal part of the flap while the deeper perialar region involving the upper lip was covered with the thicker distal part. The flaps have shown sufficient blood flow after the operation. There was no significant pin cushioning or "trap-door" effect in any case. Mild erythema and edema was found in few cases. The patients were satisfied with the cosmetic result of the intervention. Conclusions: The flap is suitable for the reconstruction of alar defects involving the perialar region. It has the advantage of covering the deeper perialar and the thinner alar defects, whilst eliminating the pin cushioning effect of the conventional subcutaneous island pedicle flaps.

Ryanodine receptors have an important role in the regulation of intracellular calcium levels in the nervous system and muscle. It has been described that ryanodine receptors influence keratinocyte differentiation and barrier homeostasis. Our goal was to examine the role of ryanodine receptors in the healing of full-thickness dermal wounds by means of in vitro and in vivo methods. The effect of ryanodine receptors on wound healing, microcirculation and inflammation was assessed in an in vivo mouse wound healing model, using skin fold chambers in the dorsal region, and in HaCaT cell scratch wound assay in vitro. SKH-1 mice were subjected to sterile saline (n = 36) or ryanodine receptor agonist 4-chloro-m-cresol (0.5 mM) (n = 42) or ryanodine receptor antagonist dantrolene (100 μM) (n = 42). Application of ryanodine receptor agonist 4-chloro-m-cresol did not influence the studied parameters significantly, whereas ryanodine receptor antagonist dantrolene accelerated the wound closure. Inhibition of the calcium channel also increased the vessel diameters in the wound edges during the process of healing and increased the blood flow in the capillaries at all times of measurement. Furthermore, application of dantrolene decreased xanthine-oxidoreductase activity during the inflammatory phase of wound healing. Inhibition of ryanodine receptor-mediated effects positively influence wound healing. Thus, dantrolene may be of therapeutic potential in the treatment of wounds.

Background Glycerol is known to possess anti-irritant and hydrating properties and previous studies suggested that xylitol may also have similar effects. Objective Our aim was to study whether different concentrations of these polyols restore skin barrier function and soothe inflammation in sodium lauryl sulphate (SLS)-induced acute irritation. Methods The experiments were performed on male SKH-1 hairless mice. The skin of the dorsal region was exposed to SLS (5%) for 3 h alone or together with 5% or 10% of glycerol respectively. Further two groups received xylitol solutions (8.26% and 16.52% respectively) using the same osmolarities, which were equivalent to those of the glycerol treatments. The control group was treated with purified water. Transepidermal water loss (TEWL) and skin hydration were determined. Microcirculatory parameters of inflammation were observed by means of intravital videomicroscopy (IVM). Furthermore, accumulation of neutrophil granulocytes and lymphocytes, the expression of inflammatory cytokines and SLS penetration were assessed, as well. Results Treatment with the 10% of glycerol and both concentrations of xylitol inhibited the SLS-induced elevation of TEWL and moderated the irritant-induced increase in dermal blood flow and in the number of leucocyte-endothelial interactions. All concentrations of the applied polyols improved hydration and prevented the accumulation of lymphocytes near the treatment site. At the mRNA level, neither glycerol nor xylitol influenced the expression of interleukin-1 alpha. However, expression of interleukin-1 beta was significantly decreased by the 10% glycerol treatment, while expression of tumour necrosis factor-alpha decreased upon the same treatment, as well as in response to xylitol. Higher polyol treatments decreased the SLS penetration to the deeper layers of the stratum corneum. Conclusion Both of the analysed polyols exert considerable anti-irritant and anti-inflammatory properties, but the effective concentration of xylitol is lower than that of glycerol.

Potassium acyltrifluoroborates (KATs) are fascinating functional groups whose further exploration is limited by poor synthetic access. Documented herein is the design and synthesis of a new reagent for their one-step preparation from aryl- and heteroarylhalides. The reagent is a stable, soluble zwitterion prepared by S-alkylation of a novel thioformamide trifluoroboronate. The KATs are prepared by adding one equivalent of nBuLi to a mixture of the aryl halide and the reagent at -78 °C. This protocol is suitable for the preparation of KATs containing pyridines, esters, nitro groups, and halides.

Cross-linking of the hyaluronic acid (hyaluronan, HA) molecule may be a possible chemical modification via which to improve the problem of the penetration of HA. The successful preparation and characterization of stable nanoparticulate systems based on HA were described and discussed previously. The purpose of the present study is to compare hydrogels based on linear or cross-linked HA as potential semisolid drug delivery forms from the aspects of deep HA penetration through the skin. The rheological properties, hydration, irritation effect, in vitro and in vivo skin penetration were studied. The hydration effect was kept and the rheological parameters were slightly changed after the cross-linking procedure. Diffusion and penetration studies demonstrated that the formation of smaller particles of HA by means of cross-linking resulted in better diffusion through a synthetic membrane and better penetration through the human epidermis and living animal skin as compared with linear HA, which did not penetrate.

Dermal or transdermal medication may lead to irritant contact dermatitis. However, little information is available on the irritant effect of surfactants which are applied in topical formulations. Our aim was to examine the irritant effect of the most frequent compounds in topical products. A murine model was applied. The following compounds were examined: sodium lauryl sulphate (SLS), polyethoxylated (40EO) hydrogenated castor oil and sucrose laurate. SLS led to severe erythema, increase in transepidermal water loss (TEWL) and induced necrosis and accumulation of neutrophylic granulocytes and lymphocytes. Exposure to sucrose laurate resulted in an elevation of TEWL, but histology did not reveal impairment of the skin structure. Application of polyethoxylated (40EO) hydrogenated castor oil was not accompanied by tissue damage. Special attention should be paid to the irritant effect of SLS. Polyethoxylated (40EO) hydrogenated castor oil seems to be a non-irritant agent and sucrose laurate is also a promising candidate for application in topical preparations.

The present paper describes the preparation and characterization of hydrophilic hyaluronan nanoparticles formed by cross-linking of hyaluronic acid (HA) with 2,2′(ethylenedioxy)bis(ethylamine) in the presence of water-soluble carbodiimide (CDI) in aqueous media. The particle size of cross-linked nanosystems was measured by dynamic light scattering (DLS) and transmission electron microscopy (TEM), transmittance by UV–VIS spectrophotometry, gel permeation chromatography (GPC) and rheology to characterize the physico-chemical properties. The aqueous nanosystems prepared were stable, transparent or mildly opalescent with values of transmittance above 91%. It was observed, that a fraction of particles with size less than 20 nm, has been released during the purification by diafiltration. It was established that the hydrodynamic size of cross-linked HA nanoparticles can be controlled by variation of the reaction conditions such as concentration of HA, salt concentration of media, ratio of cross-linker and the final pH of the reaction mixture.

The aim of this study was to develop transdermal gel formulations for ibuprofen, which ensure good skin permeation into the deeper layers, hereby achieving effective pain and inflammation relief locally. Transcutol and a new generation surfactant, a sucrose ester, were used as penetration enhancer. Ibuprofen diffusion was investigated across synthetic membrane (in vitro), and permeation was examined through excised human epidermis (ex vivo) and hairless mice (in vivo) too. Our investigations revealed that Transcutol is an effective diffusion increaser for ibuprofen, but it could not enhance its skin permeation. However, the sucrose ester promoted skin permeation of ibuprofen 2.15 fold. From our study, it seems that it is not enough to make in vitro membrane diffusion measurements by testing newly developed transdermal preparations, but it is also indispensable to complete the examinations with ex vivo skin permeation method. Our investigations show that sucrose laurate seems to be an appropriate and effective penetration and permeation enhancer for ibuprofen.

Blanka Sütö,1 Szilvia Berkó,1 Gábor Kozma,2 Ákos Kukovecz,2,3 Mária Budai-Szucs,1 Gábor Erös,4,5 Lajos Kemény,4 Anita Sztojkov-Ivanov,6 Róbert Gáspár,6 Erzsébet Csányi11Department of Pharmaceutical Technology, Faculty of Pharmacy, 2Department of Applied and Environmental Chemistry, 3MTA-SZTE “Lendület” Porous Nanocomposites Research Group, 4Department of Dermatology and Allergology, 5Department of Oral Biology and Experimental Dental Research, 6Department of Pharmacodynamics and Biopharmacy, University of Szeged, Szeged, HungaryAbstract: An ibuprofen-loaded nanostructured lipid carrier (IBU-NLC) was developed for enhanced skin penetration to improve the treatment of osteoarthritis and other musculoskeletal diseases. The mean particle size was 106nm, with a spherical morphology, a smooth surface, and a zeta potential of −18.4mV. X-ray diffraction studies revealed the amorphous state of the lipid matrix. Both Raman spectroscopy and Fourier transformation infrared analysis indicated no major shifts in the spectra of the formulations, which suggest rapid drug dissolution from the nanoparticles. The drug loading was 9.85%, and the entrapment efficiency was 98.51%. In vitro release of the NLC dispersion, in vitro permeation, and in vivo animal studies of IBU-NLC gel all confirmed that the permeation of IBU was significantly better than that of a reference after 6hours. In conclusion, IBU-NLC gel is of great potential to enhance drug permeation through the skin and hence the efficacy of the treatment of chronic joint inflammation.Keywords: ibuprofen, nanostructured lipid carriers, skin penetration, SKH-1 hairless mice, osteoarthritis

The aim of the present work was the optimization of the transdermal delivery of a lyotropic liquid crystal genistein-based formulation (LLC-GEN). LLC was chosen as medium in view of the poor solubility of GEN in water. Membrane diffusion and penetration studies were carried out with a Franz diffusion cell, through a synthetic membrane in vitro, a chick chorioallantoic membrane ex ovo, and ex vivo excised human epidermis. Thereafter, LLC-GEN was combined with electroporation (EP) to enhance the transdermal drug delivery. The synergistic effect of EP was verified by in vivo ATR-FTIR and ex vivo Raman spectroscopy on hairless mouse skin.

The identification of fast, chemoselective bond-forming reactions is one of the major contemporary challenges in chemistry. We show that chemoselective amide-forming ligations of potassium acyltrifluoroborates (KATs) and O-carbamoylhydroxylamines proceed in the presence of all unprotected functional groups with a second-order rate constant of 20 M(-1) s(-1). PEG chains, lipids, biotin, and dyes were introduced onto an unprotected 31-mer peptide (a GLP-1 analogue) with equimolar ratios of reactants within minutes at 1 mM and within 1 h at 100 μM, even with Mw 20,000 PEG. This conjugation reaction provides a new approach to the synthesis of molecules such as protein-protein and protein-polymer conjugates.

Aerobic methane generation was demonstrated earlier in plants and eukaryotes under various stress conditions. Our aims were to develop a real-time and noninvasive detection system for monitoring the methane production of small animals and humans with our without exposure to various treatments. A near-infrared diode laser technique was employed with photoacoustic spectroscopy to monitor a methane-containing atmosphere online. The whole-body methane generation of anesthetized mice and rats was determined under baseline conditions and following reduction of the intestinal methanogenic flora or after lipopolysaccharide administration. Single-breath methane analyses were also carried out in a cross-sectional clinical study in order to obtain comparative human data. The whole-body methane production of mice was significantly decreased after antibiotic treatment (M: 1.71 ppm cm(-2) 10(3); p25: 1.5 ppm cm(-2) 10(3); p75: 2.11 ppm cm(-2) 10(3)) and increased significantly in endotoxemia (M: 4.53 ppm cm(-2) 10(3); p25: 4.37 ppm cm(-2) 10(3); p75: 5.38 ppm cm(-2) 10(3)), while no difference was observed between the rat groups. The methane content of the exhaled breath in humans was found to be between 0 and 37 ppm. Photoacoustic spectroscopy is a reliable tool with which to monitor the in vivo dynamics of stress-induced methane production in laboratory animals, even in a very low concentration range.

Analysis of breath and gases emanated from skin can be used for early and non-invasive diagnosis of various kinds of diseases. Two portable, compact, photoacoustic spectroscopy based trace gas sensors were developed for the detection of methane emanated from skin and ammonia emanated from oral cavity. The light sources were distributed feedback diode lasers emitting at the absorption lines of ammonia and methane, at 1.53 μm and 1.65 μm, respectively. Photoacoustic method ensures high selectivity, therefore cross-sensitivity was negligible even with large amount of water vapor and carbon dioxide in the gas sample. In case of ammonia a preconcentration unit was used to achieve lower minimum detectable concentration. Gas sample from the oral cavity was drawn through a glass tube to the preconcentration unit that chemically bonded ammonia and released it when heated. The minimum detectable concentration of ammonia was 10 ppb for 15 s gas sampling time (gas sample of 250 cm 3 ). For methane minimum detectable concentration of 0.25 ppm was found with 12 s integration time, and it was proved to be adequate for the detection of methane emanated from human skin and from mice. Instruments measuring methane and ammonia are currently installed at two medical research laboratories at University of Szeged and tested as instruments for non-invasive clinical trials. The aim of the measurements is to determine correlations between diseases or metabolic processes and emanated gases.

Background Joints are privileged compartments that enjoy increased protection against the inflammatory reactions affecting the extremities. We hypothesized that the functional characteristics of the microvasculature would contribute to the differential defensive potential of the synovial membrane. Methods We investigated the synovial microcirculatory reactions and compared them with those of the tibial periosteum in response to 60 min of total limb ischemia, followed by 180 min of ischemia–reperfusion (IR) in rats. Carrageenan/kaolin-induced knee monoarthritis, a neutrophil-driven synovial inflammation model, served as the positive control. Results IR brought about a significant reduction in red blood cell velocity in the capillaries and increases in rolling and adherence of the neutrophil leukocytes in the postcapillary venules (intravital microscopy), in adhesion molecule expression (intercellular adhesion molecule-1 immunohistochemistry) and in xanthine oxidoreductase activity in the periosteum. These changes were also pronounced in carrageenan/kaolin-induced monoarthritis but were almost completely absent in the synovium after the IR challenge. Most importantly, even after IR and in carrageenan/kaolin monoarthritis, the synovial microcirculation was characterized by significantly greater red blood cell velocities than that in the periosteum under resting conditions. Conclusions The ischemic duration, which significantly affected the functional integrity of the periosteal microcirculation, did not bring about a marked deterioration in that of the synovial membrane, suggesting that the synovial microcirculation is less endangered to the consequences of short-term tourniquet exposure than the periosteum. The greater microcirculatory red blood cell velocities and lower IR-induced endothelial expression of intercellular adhesion molecule-1 in the synovial membrane might explain the greater resistance of this compartment to the inflammatory consequences of IR.