Evaluation of a protocol to assess a novel artificial biofilm equivalent for dentures-A prospective clinical pilot study.

OBJECTIVES: This pilot study aimed to carry out preliminary tests of the removability of an artificial biofilm equivalent (ABE) and to verify the reproducibility of the ABE testing protocol for a planned main study. BACKGROUND: There is a lack of data to develop suitable artificial biofilm substitutes, which may be helpful to perform denture hygiene education and to carry out in vitro examinations of oral hygiene products. MATERIALS AND METHODS: This single-group, prospective, longitudinal, interventional pilot study was conducted in Dresden (Germany) from February until December 2020. Participants were recruited who wore fully functional upper complete dentures. Denture biofilm was grown on acrylic specimens by wearing dentures for 12 h and 36 h using intraoral appliances. Acrylic specimens were coated with ABEs of three compositions: chitosan (ChS) 0.3 g, methylcellulose (MC) 1.7 g; ChS 0.2 g, MC 1.8 g; ChS 0.1 g, MC 1.9 g (labelled 1.7MC, 1.8MC and 1.9MC, respectively). All specimens underwent standardised mechanical brushing. The percentages of remaining biofilm (POB) were measured. RESULTS: Thirty-one participants were prescreened, and eight (26%) were included. The appliances were well tolerated, and biofilm was collected. ABE was prepared and brushed as planned. Three and six brushing strokes were needed to remove 12-h and 36-h natural denture biofilm, respectively. Correspondingly, three brushing strokes were needed to remove 1.9MC ABE and six brushing strokes to remove 1.8MC and 1.7MC ABE. A reproducibility of ABE removal was indicated. CONCLUSION: The removability of ABE and the ABE testing protocol were feasible and reproducible for conducting the future main study.

Selected miRNA and Psoriasis-Cardiovascular Disease (CVD)-Overweight/Obesity Network-A Pilot Study.

Psoriasis is nowadays recognized as a multifactorial systemic disease with complex and not fully understood pathogenesis. In psoriatic patients, the increased cardiovascular disease (CVD) risk and frequent comorbidities like obesity are observed. The aim of this study was to investigate differences in miRNA (miR-22-3p, miR-133a-3p, miR-146a-5p, miR-369-3p, and Let-7b-5p) involved in CVD risk among psoriatic patients with overweight/obesity and with normal weight. The study comprised 28 male psoriatic patients and 16 male healthy controls. miRNA isolated from peripheral blood mononuclear cells was reverse-transcribed and RT-qPCR was performed. We have found decreased levels of miR-22, miR-133a, miR-146a, and miR-369 among the psoriatic patients. There was a statistically significant difference in miR-22 and miR-146a levels between psoriatic patients with overweight/obesity and with normal weight. There were positive correlations between miR-22 and miR-146a levels and psoriatic arthritis (PsA) in psoriatic patients with normal weight and between the miR-133a level and PsA in the overweight/obese patients. The decreased levels of selected miRNA are consistent with the levels observed in CVD indicating their impact on the CVD risk in psoriatic patients. miR-22 and miR-146 may be recognized as one of the contributing factors in the obesity-CVD-psoriasis network.

The biological interactions between kynurenine and AhR in melanocytes: in vitro studies.

Kynurenine (KYN), a tryptophan metabolite, is endogenously produced by the skin cells and is present in human sweat. The aim of this study was to determine the molecular mechanism of the antiproliferative activity of KYN on human epidermal melanocytes. KYN significantly inhibited the metabolic activity of HEMa cells by decreasing cyclin D1 and cyclin-dependent kinase 4 (CDK4) levels via the aryl hydrocarbon receptor (AhR) pathway. The results suggested that KYN might be involved in the regulation of physiological and pathological processes mediated by melanocytes.

Serum copeptin as a predictor of risk of hyponatremia after transurethral prostatectomy.

BACKGROUND: Transurethral resection of the prostate gland (TURP) frequently leads to the development of dilutional hyponatremia. Copeptin has been established as a surrogate marker of vasopressin and is measured for clinical assessment of various sodium and water disturbances. This study aims to assess the utility of serum concentration of copeptin and N-terminal prohormone of brain natriuretic peptide (NT-proBNP) for prediction of post-TURP alterations of serum sodium concentration. METHODS: Forty-three patients with benign prostatic hyperplasia undergoing TURP were enrolled. Serum sodium and copeptin were measured before the procedure, then 12 hours after its completion. NT-proBNP was assessed at baseline. The total amount of fluids and sodium administered intravenously and used to flush the bladder during TURP was calculated in each patient. Receiver operator characteristic (ROC) curve analysis was used to determine value of copeptin and NT-proBNP for prediction of hyponatremia after TURP. RESULTS: In forward stepwise multiple regression analysis of serum copeptin before surgery and the duration of TURP explained the significant portion of the sodium concentration variation 12 hours from the start of the surgery. ROC curve analysis showed that serum copeptin before surgery predicted development of hyponatremia 12 hours after TURP (area under the curve, 0.775; 95% confidence interval, 0.62-0.89; p < 0.001) with a cut-off point of >78.6 pg/mL with 77% sensitivity and 64.7% specificity. Serum NT-proBNP before surgery did not predict hyponatremia 12 hours after TURP. CONCLUSION: Serum copeptin before TURP surgery, but not NT-proBNP, may be a clinically useful marker of the risk of serum sodium decrease after TURP.

PPARs and the Kynurenine Pathway in Melanoma-Potential Biological Interactions.

Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors involved in various physiological and pathological processes within the skin. PPARs regulate several processes in one of the most aggressive skin cancers, melanoma, including proliferation, cell cycle, metabolic homeostasis, cell death, and metastasis. In this review, we focused not only on the biological activity of PPAR isoforms in melanoma initiation, progression, and metastasis but also on potential biological interactions between the PPAR signaling and the kynurenine pathways. The kynurenine pathway is a major pathway of tryptophan metabolism leading to nicotinamide adenine dinucleotide (NAD+) production. Importantly, various tryptophan metabolites exert biological activity toward cancer cells, including melanoma. Previous studies confirmed the functional relationship between PPAR and the kynurenine pathway in skeletal muscles. Despite the fact this interaction has not been reported in melanoma to date, some bioinformatics data and biological activity of PPAR ligands and tryptophan metabolites may suggest a potential involvement of these metabolic and signaling pathways in melanoma initiation, progression, and metastasis. Importantly, the possible relationship between the PPAR signaling pathway and the kynurenine pathway may relate not only to the direct biological effect on melanoma cells but also to the tumor microenvironment and the immune system.

Comparison of immunohistochemical and qPCR methods from granulomatous dermatitis lesions for detection of leishmania in dogs living in endemic areas: a preliminary study.

BACKGROUND: In canine leishmaniosis (CanL) endemic areas, pathologists often receive skin biopsies for testing with histopathologic findings suggestive-but not conclusive for a definitive diagnosis-of CanL lesions. I the absence of data on the infective status of animals, the diagnosis can therefore be challenging. The aim of this retrospective study was to evaluate the ability of immunohistochemistry (IHC) and quantitative PCR (qPCR) methods to detect Leishmania infection in skin biopsies with a histopathologic diagnosis of lymphoplasmacytic/histiocytic and/or granulomatous dermatitis and to correlate the pattern, depth and severity of the histopathologic lesions with the parasite load detected by qPCR and IHC. METHODS: Thirty formalin-fixed, paraffin-embedded skin samples were evaluated by hematoxylin-eosin (H&E) staining, IHC, conventional PCR (cPCR) and qPCR. The severity, pattern and depth of the dermal inflammation and parasite load were graded. RESULTS: Leishmania was detected by H&E staining in 8/30 sections (26.66%) and by IHC in 14/30 samples (46.66%). Parasite DNA was detected in 14/30 samples (46.66%) by cPCR and in 21/30 samples (70%) by qPCR, with an extremely variable parasite load (1.32-62.700 copies). The level of agreement was fair between H&E staining and cPCR (κ = 0.32), and moderate between H&E staining and IHC (κ = 0.58). The level of agreement between IHC and cPCR was good (κ = 0.65); between IHC and qPCR, moderate (κ = 0.41); and between cPCR and qPCR, fair (κ = 0.28). A significant association was found between the severity of dermal inflammation and the parasitic skin load by IHC, although with weak linear correlation. CONCLUSIONS: Our study underlines the difficulty of obtaining a definitive diagnosis of CanL cutaneous lesions, even with the most accurate diagnostic tests currently available. Based on our results, no single test is suitable on its own for the diagnosis of cutaneous lesions caused by Leishmania. However, in the presence of a moderate/severe lymphoplasmacytic/histiocytic and/or granulomatous dermatitis, we suggest performing IHC, as in our study this technique proved to be the method with the highest discriminatory power to estimate the role of the parasite in skin lesions. In mild lesions, IHC loses its discriminatory power and should be effectively combined with techniques such as qPCR.

Modification of TiAlV Alloys with Hybrid Layers Containing Metallic Nanoparticles Obtained by the Sol-Gel Method: Surface and Structural Properties.

The aim of the work was to obtain hybrid coatings containing silver, copper, and zinc nanoparticles on the TiAlV medical alloy via a sol-gel process. The developed layers were designed to bring about a bactericidal and fungicidal effect, as well as for protection against surgical scratches during the implantation of implants used in veterinary medicine. In this work, the authors focused on evaluating the microstructure (SEM + EDS); the structure (XRD, FTIR); and the surface properties, such as wettability, free surface energy, and roughness of layers with various concentrations of metallic nanoparticles (2 and 5 mol %). Our results confirmed that the sol-gel method enables the easy manufacturing of hybrid layers endowed with different porosity values as well as various shapes and sizes of metallic nanoparticles. A higher concentration of nanoparticles was observed on the surface containing 5 mol % of metallic salts. The highest degree of homogeneity was obtained for the layers containing silver nanoparticles. In addition, the silver nanoparticles were round and had the smallest dimensions, even below 20 nm. The FTIR and XRD structural studies confirmed the presence of an organosilicon matrix containing all three types of the metallic particles. We conclude that the higher concentration of nanoparticles influenced the alloy surface parameters.

Hydrogen Storage in Pure and Boron-Substituted Nanoporous Carbons-Numerical and Experimental Perspective.

Nanoporous carbons remain the most promising candidates for effective hydrogen storage by physisorption in currently foreseen hydrogen-based scenarios of the world's energy future. An optimal sorbent meeting the current technological requirement has not been developed yet. Here we first review the storage limitations of currently available nanoporous carbons, then we discuss possible ways to improve their storage performance. We focus on two fundamental parameters determining the storage (the surface accessible for adsorption and hydrogen adsorption energy). We define numerically the values nanoporous carbons have to show to satisfy mobile application requirements at pressures lower than 120 bar. Possible necessary modifications of the topology and chemical compositions of carbon nanostructures are proposed and discussed. We indicate that pore wall fragmentation (nano-size graphene scaffolds) is a partial solution only, and chemical modifications of the carbon pore walls are required. The positive effects (and their limits) of the carbon substitutions by B and Be atoms are described. The experimental 'proof of concept' of the proposed strategies is also presented. We show that boron substituted nanoporous carbons prepared by a simple arc-discharge technique show a hydrogen adsorption energy twice as high as their pure carbon analogs. These preliminary results justify the continuation of the joint experimental and numerical research effort in this field.

UVB Radiation and Selected Tryptophan-Derived AhR Ligands-Potential Biological Interactions in Melanoma Cells.

Excessive UV exposure is considered the major environmental factor in melanoma progression. Human skin is constantly exposed to selected tryptophan-derived aryl hydrocarbon receptor (AhR) ligands, including kynurenine (KYN) and kynurenic acid (KYNA), as they are endogenously produced and present in various tissues and body fluids. Importantly, recent studies confirmed the biological activity of KYN and KYNA toward melanoma cells in vitro. Thus, in this study, the potential biological interactions between UVB and tryptophan metabolites KYN and KYNA were studied in melanoma A375, SK-MEL-3, and RPMI-7951 cells. It was shown that UVB enhanced the antiproliferative activity of KYN and KYNA in melanoma cells. Importantly, selected tryptophan-derived AhR ligands did not affect the invasiveness of A375 and RPMI-7951 cells; however, the stimulatory effect was observed in SK-MEL-3 cells exposed to UVB. Thus, the effect of tryptophan metabolites on metabolic activity, cell cycle regulation, and cell death in SK-MEL-3 cells exposed to UVB was assessed. In conclusion, taking into account that both UVB radiation and tryptophan-derived AhR ligands may have a crucial effect on skin cancer formation and progression, these results may have a significant impact, revealing the potential biological interactions in melanoma cells in vitro.

Kynurenine emerges from the shadows - Current knowledge on its fate and function.

Kynurenine (KYN), a main metabolite of tryptophan in mammals, is a direct precursor of kynurenic acid, anthranilic acid and 3-hydroxykynurenine (3-HK). Under physiological conditions, KYN is produced endogenously mainly in the liver by tryptophan 2,3-dioxygenase (TDO). Tumorigenesis and inflammatory conditions increase the activity of another KYN synthetizing enzyme, indoleamine 2,3-dioxygenase (IDO). However, knowledge about the exogenous sources and the fate of KYN in mammals is still limited. While most papers deal with the contribution of KYN to pathologies of the central nervous system, its role in the periphery has almost been ignored. KYN is a ligand for the aryl hydrocarbon receptor (AhR). As a receptor for KYN and its downstream metabolites, AhR is involved in several physiological and pathological conditions, including inflammation and carcinogenesis. Recent studies have shown that KYN suppresses immune response and is strongly involved in the process of carcinogenesis and tumour metastasis. Thus, inhibition of activity of the enzymes responsible for KYN synthesis, TDO, IDO or genetic manipulation leading to reduction of KYN synthesis, could be considered as innovative strategies for improving the efficacy of immunotherapy. Surprisingly, however, genetic or pharmacological approaches for reducing tryptophan catabolism to KYN do not necessarily result in decrease of KYN level in the main circulation. This review aims to summarize the current knowledge of KYN fate and function and to emphasize its importance for vital physiological and pathological processes.

A New Insight into the Potential Role of Tryptophan-Derived AhR Ligands in Skin Physiological and Pathological Processes.

The aryl hydrocarbon receptor (AhR) plays a crucial role in environmental responses and xenobiotic metabolism, as it controls the transcription profiles of several genes in a ligand-specific and cell-type-specific manner. Various barrier tissues, including skin, display the expression of AhR. Recent studies revealed multiple roles of AhR in skin physiology and disease, including melanogenesis, inflammation and cancer. Tryptophan metabolites are distinguished among the groups of natural and synthetic AhR ligands, and these include kynurenine, kynurenic acid and 6-formylindolo[3,2-b]carbazole (FICZ). Tryptophan derivatives can affect and regulate a variety of signaling pathways. Thus, the interest in how these substances influence physiological and pathological processes in the skin is expanding rapidly. The widespread presence of these substances and potential continuous exposure of the skin to their biological effects indicate the important role of AhR and its ligands in the prevention, pathogenesis and progression of skin diseases. In this review, we summarize the current knowledge of AhR in skin physiology. Moreover, we discuss the role of AhR in skin pathological processes, including inflammatory skin diseases, pigmentation disorders and cancer. Finally, the impact of FICZ, kynurenic acid, and kynurenine on physiological and pathological processes in the skin is considered. However, the mechanisms of how AhR regulates skin function require further investigation.

Antifungal and Surface Properties of Chitosan-Salts Modified PMMA Denture Base Material.

Chitosan (CS) and its derivatives show antimicrobial properties. This is of interest in preventing and treating denture stomatitis, which can be caused by fungi. Therefore, the aim of this study was the development of a novel antifungal denture base material by modifying polymethyl methacrylate (PMMA) with CS-salt and characterizing its antifungal and surface properties in vitro. For this purpose, the antifungal effect of chitosan-hydrochloride (CS-HCl) or chitosan-glutamate (CS-G) as solutions in different concentrations was determined. To obtain modified PMMA resin specimens, the CS-salts were added to the PMMA before polymerization. The roughness of these specimens was measured by contact profilometry. For the evaluation of the antifungal properties of the CS-salt modified resins, a C. albicans biofilm assay on the specimens was performed. As solutions, both the CS-G and CS-HCl-salt had an antifungal effect and inhibited C. albicans growth in a dose-dependent manner. In contrast, CS-salt modified PMMA resins showed no significant reduced C. albicans biofilm formation. Furthermore, the addition of CS-salts to PMMA significantly increased the surface roughness of the specimens. This study shows that despite the antifungal effect of CS-salts in solution, a modification of PMMA resin with these CS-salts does not improve the antifungal properties of PMMA denture base material.

Effect of Tryptophan-Derived AhR Ligands, Kynurenine, Kynurenic Acid and FICZ, on Proliferation, Cell Cycle Regulation and Cell Death of Melanoma Cells-In Vitro Studies.

Tryptophan metabolites: kynurenine (KYN), kynurenic acid (KYNA) and 6-formylindolo[3,2-b]carbazole (FICZ) are considered aryl hydrocarbon receptor (AhR) ligands. AhR is mainly expressed in barrier tissues, including skin, and is involved in various physiological and pathological processes in skin. We studied the effect of KYN, KYNA and FICZ on melanocyte and melanoma A375 and RPMI7951 cell toxicity, proliferation and cell death. KYN and FICZ inhibited DNA synthesis in both melanoma cell lines, but RPMI7951 cells were more resistant to pharmacological treatment. Tested compounds were toxic to melanoma cells but not to normal human adult melanocytes. Changes in the protein level of cyclin D1, CDK4 and retinoblastoma tumor suppressor protein (Rb) phosphorylation revealed different mechanisms of action of individual AhR ligands. Importantly, all tryptophan metabolites induced necrosis, but only KYNA and FICZ promoted apoptosis in melanoma A375 cells. This effect was not observed in RPMI7951 cells. KYN, KYNA and FICZ in higher concentrations inhibited the protein level of AhR but did not affect the gene expression. To conclude, despite belonging to the group of AhR ligands, KYN, KYNA and FICZ exerted different effects on proliferation, toxicity and induction of cell death in melanoma cells in vitro.

Bond strength of modern self-adhesive resin cements to human dentin and different CAD/CAM ceramics.

PURPOSE: The aim of the study was to evaluate the shear bond strength of CAD/CAM ceramics to dentin after cementation with conventional or self-adhesive resin cements. METHODS: Three self-adhesive, self-etching cements (Panavia SA, RelyX U200, Maxcem Elite), and one conventional cement (Panavia V5), were selected to lute three CAD/CAM ceramics (IPS Empress CAD, IPS e.max CAD, IPS e.max ZirCAD) onto the dentin. The bond strength was evaluated using a shear strength test according to the PN-EN ISO 29022:2013-10. Evaluation of the differences was performed using the Statistica software. Failure modes were analyzed using a light microscope. RESULTS: All the studied cements differed (regardless of the ceramic type) in the bond strength. The highest bond strength was observed in Panavia V5, lower - in RelyX U200 and Panavia SA, and the lowest - in Maxcem. For IPS e.max ZirCAD, it was observed that compared to Panavia V5, the other cements were characterized by a significantly higher bond strength. For the IPS Empress CAD and the IPS e.max CAD, Panavia V5 displayed the highest bond strength. For all the studied self-adhesive cements, the failure of adhesion between the cement and dentin was predominant mode. CONCLUSIONS: Significant differences were found in the shear bond strengths of the CAD/CAM ceramics luted to dentin using tested self-adhesive and conventional cements. The bond strength depended on the combination of ceramic and cement. The IPS e.max ZirCAD had the highest bond strength to dentin after cementation with RelyX U200, while the IPS Empress CAD and IPS e.max CAD - with Panavia V5.

A Tryptophan Metabolite, 8-Hydroxyquinaldic Acid, Exerts Antiproliferative and Anti-Migratory Effects on Colorectal Cancer Cells.

8-Hydroxyquinaldic acid, the end-metabolite of tryptophan, is well-known metal chelator; however, its role in humans, especially in cancer promotion and progression, has not been fully revealed. Importantly, 8-hydroxyquinaldic acid is the analog of kynurenic acid with evidenced antiproliferative activity towards various cancer cells. In this study, we revealed that 8-hydroxyquinaldic acid inhibited not only proliferation and mitochondrial activity in colon cancer HT-29 and LS-180 cells, but it also decreased DNA synthesis up to 90.9% for HT-29 cells and 76.1% for LS-180 cells. 8-Hydroxyquinaldic acid induced changes in protein expression of cell cycle regulators (CDK4, CDK6, cyclin D1, cyclin E) and CDKs inhibitors (p21 Waf1/Cip1, p27 Kip1), but the effect was dependent on the tested cell line. Moreover, 8-hydroxyquinaldic acid inhibited migration of colon cancer HT-29 and LS-180 cells and increased the expression of ß-catenin and E-cadherin. Importantly, antiproliferative and anti-migratory concentrations of 8-hydroxyquinaldic acid were non-toxic in vitro and in vivo. We reported for the first time antiproliferative and anti-migratory activity of 8-hydroxyquinaldic acid against colon cancer HT-29 and LS-180 cells.

Kynurenic acid and cancer: facts and controversies.

Kynurenic acid (KYNA) is an endogenous tryptophan metabolite exerting neuroprotective and anticonvulsant properties in the brain. However, its importance on the periphery is still not fully elucidated. KYNA is produced endogenously in various types of peripheral cells, tissues and by gastrointestinal microbiota. Furthermore, it was found in several products of daily human diet and its absorption in the digestive tract was evidenced. More recent studies were focused on the potential role of KYNA in carcinogenesis and cancer therapy; however, the results were ambiguous and the biological activity of KYNA in these processes has not been unequivocally established. This review aims to summarize the current views on the relationship between KYNA and cancer. The differences in KYNA concentration between physiological conditions and cancer, as well as KYNA production by both normal and cancer cells, will be discussed. The review also describes the effect of KYNA on cancer cell proliferation and the known potential molecular mechanisms of this activity.

Mechanical and Surface Properties of Resilient Denture Liners Modified with Chitosan Salts.

Chitosan (CS) and its derivatives show antibacterial and antifungal properties and could help treat and prevent denture stomatitis (DS). Mechanical and surface properties of resilient denture liners were evaluated when modified with CS salts. CS-hydrochloride (CS-HCl) and CS-glutamate (CS-G) were added to resilient denture liners Ufi Gel P and Coe-Soft at four different concentrations (0.1%, 0.2%, 0.4%, 1% w/w) from which specimens were produced, as well as a control group of each material with no added CS salt. Ten specimens per group (Ø 35 mm, height 6 mm) were manufactured. They were stored in distilled water at 37 °C for a total of 30 days (d). Shore A hardness (SHA) and surface roughness (Ra) were evaluated after 24 h (T1), 7 d (T2), 14 d (T3) and 30 d (T4). Kruskal-Wallis and U-test (Bonferroni-Holm adjusted) were used for statistical analysis (p ≤ 0.05). Ra increased significantly once CS salts were added. SHA increased significantly for some groups, but all specimens fulfilled requirements set by ISO 10139-2:2016. Modification with CS salts does not influence the mechanical properties of the modified resilient denture liners in a clinically relevant manner. Despite the increased roughness, the concept is suitable for further studies. Especially antimicrobial/antibiofilm studies are needed.

Exploring the Role of Manganese on Structural, Transport, and Electrochemical Properties of NASICON-Na3Fe2-yMny(PO4)3-Cathode Materials for Na-Ion Batteries.

Given the extensive efforts focused on protecting the environment, eco-friendly cathode materials are a prerequisite for the development of Na-ion battery technology. Such materials should contain abundant and inexpensive elements. In the paper, we present NASICON-Na3Fe2-yMny(PO4)3 (y = 0, 0.1, 0.2, 0.3, and 0.4) cathode materials, which meet these requirements. Na3Fe2-yMny(PO4)3 compounds were prepared via a solid-state reaction at 600 °C, which allowed to obtain powders with submicron particles. The presence of manganese in the iron sub-lattice inhibits phase transitions, which occurs at ∼95 and ∼145 °C in Na3Fe2(PO4)3, changing the monoclinic structure to rhombohedral and affecting the structural and transport properties. The chemical stability of Na3Fe2-yMny(PO4)3 was thus higher than that of Na3Fe2(PO4)3, and it also exhibited enhanced structural, transport, and electrochemical properties. The observed correlation between the chemical composition and electrochemical properties proved the ability to precisely tune the crystal structure of NASICONs, allowing cathode materials with more desirable properties to be designed.

Kynurenic acid as the neglected ingredient of commercial baby formulas.

The global increase in resorting to artificial nutritional formulas replacing breastfeeding has been identified among the complex causes of the obesity epidemic in infants and children. One of the factors recently recognized to influence metabolism and weight gain is kynurenic acid (KYNA), an agonist of G protein-coupled receptor (GPR35). Therefore the aim of the study was to determine the concentration of KYNA in artificial nutritional formulas in comparison with its level in human breast milk and to evaluate developmental changes in rats exposed to KYNA enriched diet during the time of breastfeeding. KYNA levels were measured in milk samples from 25 heathy breast-feeding women during the first six months after labor and were compared with 21 time-adjusted nutritional formulas. Animal experiments were performed on male Wistar rats. KYNA was administered in drinking water. The content of KYNA in human milk increases more than 13 times during the time of breastfeeding while its level is significantly lower in artificial formulas. KYNA was detected in breast milk of rats and it was found that the supplementation of rat maternal diet with KYNA in drinking water results in its increase in maternal milk. By means of the immunoblotting technique, GPR35 was evidenced in the mucosa of the jejunum of 1-day-old rats and distinct morphological changes in the jejunum of 21-day-old rats fed by mothers exposed to water supplemented with KYNA were found. A significant reduction of body weight gain of rats postnatally exposed to KYNA supplementation without changes in total body surface and bone mineral density was observed. The rat offspring fed with breast milk with artificially enhanced KYNA content demonstrated a lower mass gain during the first 21 days of life, which indicates that KYNA may act as an anti-obesogen. Further studies are, therefore, warranted to investigate the mechanisms regulating KYNA secretion via breast milk, as well as the influence of breast milk KYNA on mass gain. In the context of lifelong obesity observed worldwide in children fed artificially, our results imply that insufficient amount of KYNA in baby formulas could be considered as one of the factors associated with increased mass gain.

Development of the 1,2,4-triazole-based anticonvulsant drug candidates acting on the voltage-gated sodium channels. Insights from in-vivo, in-vitro, and in-silico studies.

The treatment of epilepsy remains difficult mostly since almost 30% of patients suffer from pharmacoresistant forms of the disease. Therefore, there is an urgent need to search for new antiepileptic drug candidates. Previously, it has been shown that 4-alkyl-5-substituted-1,2,4-triazole-3-thione derivativatives possessed strong anticonvulsant activity in a maximal electroshock-induced seizure model of epilepsy. In this work, we examined the effect of the chemical structure of the 1,2,4-triazole-3-thione-based molecules on the anticonvulsant activity and the binding to voltage-gated sodium channels (VGSCs) and GABAA receptors. Docking simulations allowed us to determine the mode of interactions between the investigated compounds and binding cavity of the human VGSC. Selected compounds were also investigated in a panel of ADME-Tox assays, including parallel artificial membrane permeability assay (PAMPA), single cell gel electrophoresis (SCGE) and cytotoxicity evaluation in HepG2 cells. The obtained results indicated that unbranched alkyl chains, from butyl to hexyl, attached to 1,2,4-triazole core are essential both for good anticonvulsant activity and strong interactions with VGSCs. The combined in-vivo, in-vitro and in-silico studies emphasize 4-alkyl-5-substituted-1,2,4-triazole-3-thiones as promising agents in the development of new anticonvulsants.