Combined hyperforin and lanicemine treatment instead of ketamine or imipramine restores behavioral deficits induced by chronic restraint stress and dietary zinc restriction in mice.

Clinical and preclinical studies show evidence that chronic stress or nutritional deficits in dietary zinc (Zn) intake may be risk factors for developing major depressive disorder (MDD). Furthermore, there may be possible links between low serum Zn levels and development of treatment-resistant depression. In the present work, we combined chronic restraint stress (CRS) and a low-zinc diet (ZnD) in mice and carried out a set of behavioral and biochemical studies. The mice were treated with four different antidepressant compounds, namely, ketamine, Ro 25-6981 (Ro), hyperforin and lanicemine (Hyp + Lan), and imipramine (IMI). We show that CRS or ZnD alone or a combination of CRS and ZnD (CRS + ZnD) induces anhedonia observed in the sucrose preference test (SPT). The behavioral effects of CRS were restored by ketamine or IMI. However, only Hyp + Lan restored the deficits in behavioral phenotype in mice subjected to CRS + ZnD. We also showed that the antidepressant-like effects observed in Hyp + Lan-treated CRS + ZnD mice were associated with changes in the morphology of the dendritic spines (restored physiological level) in the hippocampus (Hp). Finally, we studied the metabolism of ketamine and its brain absorption in CRS and CRS + ZnD mice. Our results suggest that CRS + ZnD does not alter the metabolism of ketamine to (2R,6R;2S,6S)-HNK; however, CRS + ZnD can induce altered bioavailability and distribution of ketamine in the Hp and frontal cortex (FC) in CRS + ZnD animals compared to the control and CRS groups.

Zinc Deficiency Blunts the Effectiveness of Antidepressants in the Olfactory Bulbectomy Model of Depression in Rats.

Currently used antidepressants do not always provide the desired results, and many patients suffer from treatment-resistant depression. Clinical studies suggest that zinc deficiency (ZnD) may be an important risk factor for depression and might blunt the effect of antidepressants. This study aimed to examine whether ZnD might blunt the effectiveness of antidepressants in the olfactory bulbectomy model (OB) of depression in rats. For this purpose, rats were subjected to the OB model, fed a zinc-deficient diet (3 mg Zn/kg) for 3 weeks, and finally treated with escitalopram (Esc), venlafaxine (Ven) 10 mg/kg, i.p., or combined Esc/Ven (1 mg/kg, i.p.) with zinc (5 mg/kg) for another 3 weeks. Open field (OFT), forced swim (FST), and sucrose intake (SIT) tests were used to evaluate depressive-like behavioral changes. In addition, serum, intracellular, and synaptic Zn concentrations and the level of zinc transporter (ZnT) proteins were analyzed. The OB + ZnD model induced hyperactivity in rats in the OFT, increased immobility time in the FST, and anhedonia in the SIT. Chronic treatment with Esc reduced immobility time in the FST in the OB + ZnD model. Esc/Ven +Zn increased sucrose intake in rats from the OB + ZnD group. The OB + ZnD decreased serum zinc levels and intracellular and synaptic Zn concentration in the prefrontal cortex (PFC) and cerebellum. These changes were normalized by chronic administration of Esc/Ven +Zn. Moreover, OB + ZnD decreased levels of the ZnT1 protein in the PFC and Hp and ZnT3 in Hp. Chronic administration of antidepressants did not alter the levels of ZnT proteins. The OB + ZnD model induces more depressive-like effects than either model alone. Our results show that ZnD may induce drug resistance in rats. Normalizing serum or brain zinc concentration is insufficient to reverse behavioral abnormalities caused by the OB + ZnD model. However, zinc supplementation might improve the effectiveness of antidepressants in reversing particular depression symptoms.

The influence of high-fat and high-sucrose feeding regimes on organ weight, body weight, and serum concentration of bioelements in rats.

BACKGROUND: Obesity is one of the most common diseases of civilization, and approximately 13 % of the world's adult population is obese. Obesity is defined as excessive accumulation of fat in the human body, which leads to adverse health effects such as metabolic syndrome, type 2 diabetes, and fatty liver disease (e.g., nonalcoholic liver disease). The development of obesity is accompanied by changes in the levels of certain bioelements such as copper and zinc. These elements may have an influence on the proper functioning of the central nervous system. METHOD: Fifty-six male Wistar rats (initial weight 290-300 g) were divided into seven experimental groups. They were fed with different feeding patterns (constant versus intermittent-binge access) and exposed to different diets (high sucrose versus high fat) to analyze the factors that affect the organ weight gain (pancreas, spleen, liver, testes, and kidneys) and total body weight gain. Further, zinc and copper levels in the serum of the animals were determined. The relationship between organ and body weight and serum metal concentration was analyzed by cluster and principal component analyses. RESULTS: Rats with unlimited access to high fat diet (HF) and restricted intake of high sugar diet (for 2-hours daily-HSB and every second day-IHSB) have elevated body weight in comparison to the control. However, the heaviest organ weights were recorded in the HSB rats compared to the control group (pancreas, 14 %; spleen, 9 %; kidneys, 5 %; and liver, 3 %). On the other hand, an average 20 % decrease in zinc concentration was observed in rats fed with high-fat diet compared to the control. Moreover, an 18 % decrease in copper levels was observed in rats that had periodic access to high-fat diet every 2 h daily and for 2 h every other day compared to the control. CONCLUSION: Both the high-sucrose and high-fat diets had an influence on body and organ weights. This study demonstrates an association between the different types of diet and the parameters investigated (body and organ weights and concentration of serum bioelements).

Yerba Mate as a Source of Elements and Bioactive Compounds with Antioxidant Activity.

Ilex paraguariensis (yerba mate) is a plant species of the holly genus Ilex native to South America from the family Aquifoliaceae and is used for the production of yerba mate infusion. The leaves of the plant are steeped in hot water to make a beverage known as mate. The present study aimed to quantify and compare the content of selected elements in dried leaves and stems of I. paraguariensis (originating from Paraguay, Argentina, and Brazil) available in the market in Poland and determine the amount of these elements and bioactive compounds that pass into the infusion prepared from them. The contents of the following antioxidant compounds were assessed: neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 4-feruloylquinic acid, isochlorogenic acid, rutoside, astragalin, caffeine, and indole derivatives. All the tested samples showed the presence of elements such as magnesium, zinc, copper, iron, and manganese. The highest antioxidant activity was determined for infusion prepared from yerba mate samples from Brazil. Drinking approximately 1 L of the infusion a day will partially cover the daily requirement of these elements and bioactive compounds. The highest content of organic compounds with antioxidant properties (phenolic compounds and caffeine) was found in yerba mate infusions from Brazil.

Extraction of selected prohealth substances from Curcuma longa and Zingiber officinale in artificial digestive juices.

BACKGROUND: The study determined the content of selected bioelements, toxic metals and phenolic acids occurring in Curcuma longa L. and Zingiber officinale Rosc. Natural raw materials in the form of powder, slices, dried and fresh rootstock, tablets and capsules were analyzed. To determine the actual bioavailability of the metals and phenolic acids present in the raw materials, they were extracted into artificial digestive juices (artificial gastric and intestinal juices). RESULTS: The maximum amounts of bioelements found in turmeric were as follows (mg kg-1 dry weight): Ca, 1911; Cu, 10.4; Fe, 248; K, 123; Mg, 1896; Mn, 181; Na, 332; Zn, 242. On the other hand, the maximum amounts of these bioelements found in ginger were as follows (mg kg-1 dry weight): Ca, 287; Cu, 7.4; Fe, 113; K, 2903; Mg, 1162; Mn, 11.3; Na, 1678; Zn, 186. It was found that bioelements from all the tested samples of raw materials were released to artificial digestive juices. CONCLUSION: The amount of substances extracted to digestive juices from the analyzed raw materials varied depending on their form. The highest content of bioelements was found in the fresh rhizomes of turmeric and ginger. On the other hand, powdered rhizome was characterized by the highest bioavailability of elements for humans. © 2021 Society of Chemical Industry.

The history, current state and perspectives of aerosol therapy.

Nebulization is a very effective method of drug administration. This technique has been popular since ancient times when inhalation of plants rich in tropane alkaloids with spasmolytic and analgesic effects was widely used. Undoubtedly, the invention of anasthesia in the 19th century had an influence on the development of this technique. It resulted in the search for devices that facilitated anasthesia such as pulveriser or hydronium. From the second half of the 21st century, when the first DPI and MDI inhalers were launched, the constant development of aerosol therapy has been noticed. This is due to the fact that nebulization, compared with other means of medicinal substance application (such as oral and intravenous routes of administration), is safer and it exhibits a positive dose/efficacy ratio connected to the reduction of the dose. It enables drugs administration through the lung and possesses very fast onset action. Therefore, various drugs prescribed in respiratory diseases (such as corticosteroids, ß-agonists, anticholinergics) are present on the market in a form of an aerosol.

A New Biotechnology Method of Bioelements' Accumulation Monitoring in In Vitro Culture of Agaricus bisporus.

Agaricus bisporus (J.E. Lange) Imbach is one the most popular species of edible mushrooms in the world because of its taste and nutritional properties. In the research, repeatability of accumulation of bioelements and biomass yield in experimentally chosen in vitro culture medium, was confirmed. The in vitro cultures were conducted on the modified Oddoux medium enriched with bioelements (Mg, Zn, Cu, Fe). The aim of the study was to create an effective method of sampling, which enabled non-invasive monitoring of metals concentrations changes in the medium, during increase of biomass in in vitro cultures. The first, indirect method of sampling was applied. The non-invasive probe (a dipper) for in vitro culture was used; hence, the highest biomass increase and metals accumulation were gained. The method also guaranteed culture sterility. The second method, a direct one, interfered the in vitro culture conditions and growth of mycelium, and as a consequence the lower biomass increase and metals' accumulation were observed. Few cases of contaminations of mycelium in in vitro cultures were observed. The proposed method of non-invasive sampling of the medium can be used to monitor changes in the concentrations of metals in the medium and their accumulation in the mycelium in natural environment. Changes in concentrations of the selected metals over time, determined by the method of atomic absorption spectrometry, made it possible to correlate the obtained results with the specific stages of A. bisporus mycelium development and to attempt to explain the mechanism of sampling metals from the liquid substrate.

Photodegradation of Bexarotene and Its Implication for Cytotoxicity.

A detailed understanding of the stability of an active pharmaceutical ingredient and a pharmaceutical dosage form is essential for the drug-development process and for safe and effective use of medicines. Photostability testing as an inherent part of stability studies provides valuable knowledge on degradation pathways and structures of products generated under UV irradiation. Photostability is particularly important for topically administered drugs, as they are more exposed to UV radiation. Bexarotene is a more recent third-generation retinoid approved by the U.S. Food and Drug Administration and the European Medicines Agency as a topically applied anticancer agent. The present study aimed to assess bexarotene photostability, including the presence of UV filters, which have been permitted to be used in cosmetic products in Europe and the USA. The bexarotene photostability testing was performed in ethanol solutions and in formulations applied on PMMA plates. The UPLC-MS/MS technique was used to determine the tested substance. The presence of photocatalysts such as TiO2 or ZnO, as well as the organic UV filters avobenzone, benzophenone-3, meradimate, and homosalate, could contribute to degradation of bexarotene under UV irradiation. Four photocatalytic degradation products of bexarotene were identified for the first time. The antiproliferative properties of the degradation products of bexarotene were assessed by MTT assay on a panel of human adherent cancer cells, and concentration-dependent growth inhibition was evidenced on all tested cell lines. The cytotoxicity of the formed products after 4 h of UV irradiation was significantly higher than that of the parent compound (p < 0.05). Furthermore non-cancerous murine fibroblasts exhibited marked concentration-dependent inhibition by bexarotene, while the degradation products elicited more pronounced antiproliferative action only at the highest applied concentration.

Disinfectants Used in Stomatology and SARS-CoV-2 Infection.

Effective disinfection is a basic procedure in medical facilities, including those conducting dental surgeries, where treatments for tissue discontinuity are also performed, as it is an important element of infection prevention. Disinfectants used in dentistry and dental and maxillofacial surgery include both inorganic (hydrogen peroxide, sodium chlorite-hypochlorite) and organic compounds (ethanol, isopropanol, peracetic acid, chlorhexidine, eugenol). Various mechanisms of action of disinfectants have been reported, which include destruction of the structure of bacterial and fungal cell membranes; damage of nucleic acids; denaturation of proteins, which in turn causes inhibition of enzyme activity; loss of cell membrane integrity; and decomposition of cell components. This article discusses the most important examples of substances used as disinfectants in dentistry and presents the mechanisms of their action with particular focus on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The search was conducted in ScienceDirect, PubMed, and Scopus databases. The interest of scientists in the use of disinfectants in dental practice is constantly growing, which results in the increasing number of publications on disinfection, sterilization, and asepsis. Many disinfectants often possess several of the abovementioned mechanisms of action. In addition, disinfectant preparations used in dental practice either contain one compound or are frequently a mixture of active compounds, which increases their range and effectiveness of antimicrobial action. Currently available information on disinfectants that can be used to prevent SARS-CoV-2 infection in dental practices was summarized.

Effect of conservation methods on the bioaccessibility of bioelements from in vitro-digested edible mushrooms.

BACKGROUND: The release of bioelements from edible mushrooms (Agaricus bisporus, Cantharellus cibarius, and Imleria badia) was examined using in vitro simulated gastrointestinal digestion to assess their health-promoting potential. The following samples were tested: fresh, frozen, dried in a food dryer, dried in the sun, and lyophilized. The samples were incubated in gastric juice (pepsin, NaCl, HCl) and in intestinal juice (NaHCO3 , pancreatin, bile salts) with the aim of verifying the bioaccessibility of the bioelements and the digestibility of mushrooms. Four bioelements that are essential for the human body were studied: Mg, Zn, Cu, and Fe. RESULTS: It was found that Mg was extracted in the highest amounts from the sun-dried A. bisporus (1.620 g kg-1 d.w.). In the case of microelements, the lyophilized fruiting bodies of I. badia released Zn in the highest quantities (0.180 g kg-1 d.w.). Lyophilization and sun-drying methods were more advantageous than other methods. Fresh material was a more valuable source of bioelements than frozen material. CONCLUSION: Our results showed that edible mushrooms have a high content of bioelements that are easily bioaccessible, which indicates their health-promoting properties. © 2020 Society of Chemical Industry.

The remediation of sulfonamides from the environment by Pleurotus eryngii mycelium. Efficiency, products and mechanisms of mycodegradation.

The objective of the study was to assess the applicability of the mycelium obtained from the in vitro cultures of nontoxic bracket fungus, Pleurotus eryngii, to sulfonamides mycodegradation. Samples containing one of the six selected sulfonamides, sulfanilamide derivatives, were incubated with the mycelium of P. eryngii for 7 and 14 days in vitro. Subsequently, change in the sulfonamide concentration was assessed in the samples using the UPLC-QTof. The transformation products were identified based on monoisotopic molecular mass and fragmentation spectra. The studied sulfonamides did not inhibit the growth of P. eryngii mycelium in the in vitro cultures. In addition, a considerable reduction of sulfonamide concentration was observed in all the incubated samples (from 73.7 ± 8.3% to 99.8 ± 0.3%). In the case of three sulfonamides, the reduction in concentration >90% occurred after 7 days of incubation. However, the transformation of sulfonamides was partially caused by their degradation to simpler organic compounds. After incubation, the products of condensation of sulfonamides with formyl, acyl, and sugar groups, and amino acid-derived compounds were identified in the samples. This indicated the partially reversible nature of the mycodegradation process.

Photostability Testing of a Third-Generation Retinoid-Tazarotene in the Presence of UV Absorbers.

Exposure of a drug to UV irradiation could affect its physicochemical properties. Hence, photostability testing is essential for topically administered drugs. Tazarotene, a receptor-selective, third-generation retinoid, is commonly used to treat acne vulgaris and psoriasis. In the present study, an in-depth analysis of the photostability of tazarotene in ethanolic solution in the presence of zinc oxide and/or titanium dioxide as well as benzophenone-type UV filters was performed. Eleven presumed products were derived from the photocatalytic degradation of tazarotene using ultra-performance liquid chromatography-tandem mass spectrometry, and transformation pathways were proposed. The degradation process mainly affected the 4,4-dimethyl-3,4-dihydro-2H-thiopyran moiety. The fragments most susceptible to oxidation were the methyl groups and the sulfur atom. Moreover, in the presence of sulisobenzone, under UV irradiation, tazarotene was subjected to a degradation process, which resulted in two photodecomposition products. In silico studies performed by OSIRIS Property Explorer demonstrated that five of the degradation products could be harmful in terms of the reproductive effects, which are associated with 3,4-dihydro-6-methyl-2H-1-benzothiopyran 1,1-dioxide, while one of them demonstrated potential irritant activity. The cytotoxic properties of the degradation products of tazarotene were assessed by MTT assay on a panel of human adherent cancer cells. Time- and concentration-dependent growth inhibition was evidenced in ovary (A2780) and breast (MDA-MB-231) cancer cell lines. The potential implication of the outcomes of the present research requires further studies mainly concerning the photostability of tazarotene in the topical formulations.

Imipramine Influences Body Distribution of Supplemental Zinc Which May Enhance Antidepressant Action.

Zinc (Zn) was found to enhance the antidepressant efficacy of imipramine (IMI) in human depression and animal tests/models of depression. However, the underlying mechanism for this effect remains unknown. We measured the effect of intragastric (p.o.) combined administration of IMI (60 mg/kg) and Zn (40 mg Zn/kg) in the forced swim test (FST) in mice. The effect of Zn + IMI on serum, brain, and intestinal Zn concentrations; Zn transporter (ZnT, ZIP) protein levels in the intestine and ZnT in the brain; including BDNF (brain-derived neurotrophic factor) and CREB (cAMP response element-binding protein) protein levels in the brain were evaluated. Finally, the effect of IMI on Zn permeability was measured in vitro in colon epithelial Caco-2 cells. The co-administration of IMI and Zn induced antidepressant-like activity in the FST in mice compared to controls and Zn or IMI given alone. This effect correlated with increased BDNF and the ratio of pCREB/CREB protein levels in the prefrontal cortex (PFC) compared to the control group. Zn + IMI co-treatment increased Zn concentrations in the serum and brain compared to the control group. However, in serum, co-administration of IMI and Zn decreased Zn concentration compared to Zn alone treatment. Also, there was a reduction in the Zn-induced enhancement of ZnT1 protein level in the small intestine. Zn + IMI also induced an increase in the ZnT4 protein level in the PFC compared to the control group and normalized the Zn-induced decrease in the ZnT1 protein level in the hippocampus (Hp). The in vitro studies revealed enhanced Zn permeability (observed as the increased transfer of Zn through the intestinal cell membrane) after IMI treatment. Our data indicate that IMI enhances Zn transfer through the intestinal tract and influences the redistribution of Zn between the blood and brain. These mechanisms might explain the enhanced antidepressant efficacy of combined IMI/Zn treatment observed in the FST in mice.

Feasibility of the use of Lentinula edodes mycelium in terbinafine remediation.

A detailed understanding of the fate of xenobiotics introduced into the environment and the mechanisms involved in their biotransformation, biodegradation, and biosorption is essential to improve the efficiency of remediation techniques. Mycoremediation is a form of bioremediation technique that has become increasingly popular in recent years as fungi are known to produce various effective extracellular enzymes that have the potential to neutralize a wide variety of xenobiotics released into the environment. Hence, mycoremediation appears to be a promising technique for the removal of a wide array of toxins and pharmaceutical residues from a damaged environment and wastewater. This study primarily aimed to investigate whether white-rot fungus (Lentinula edodes) can be utilized for the bioremediation of common antifungal agent terbinafine, which is mainly available in the market as powder or cream. The cultures of L. edodes were cultivated in the medium containing terbinafine powder or terbinafine 1% cream, each at a final concentration of 0.1 mg mL-1. The addition of terbinafine in powder form have a negative effect on biomass growth (p < 0.05). The total amount of terbinafine in the dry weight of mycelium after culture was estimated to be 7.63 ± 0.45 mg and 12.52 ± 2.46 mg for powder and cream samples, respectively. In addition, there were no traces of terbinafine in any of the samples of medium used for culturing L. edodes after the experimental duration period. The biodegradation products of terbinafine were identified for the first time using UPLC/MS/MS. The biodegradation of terbinafine resulted in the loss of 1-naphthylmethanol, which occurred via oxidative deamination, N-demethylation, or tert-butyl group hydroxylation. The results of the study demonstrate that L. edodes mycelium can be effectively used for the remediation of terbinafine.

Pleurotus spp. Mycelia Enriched in Magnesium and Zinc Salts as a Potential Functional Food.

Worldwide, mushrooms belonging to the Pleurotus spp. such as P. citrinopileatus, P. djamor, and P. pulmonarius are highly valued not only for their taste and aroma but also for their health-promoting properties. These species are rich in bioelements, vitamins, and above all, compounds that exhibit immunostimulatory activity. Therefore, in this study, we aimed to determine the effect of the supplementation of culture media using inorganic Mg and Zn salts. This is the first study to establish the bioavailability of the selected elements (Mg and Zn) and anions (Cl-, SO42-) from the enriched biomass by means of the extraction of lyophilized mycelium into artificial digestive juices. The following salts were added to the liquid Oddoux medium: ZnSO4·7H2O, ZnCl2, MgSO4·7H2O or MgCl2·6H2O. The bioelements, anions and organic compounds in the obtained biomass were determined. The addition of Zn and Mg salts to the media increased the production of biomass by 30% and increased the bioaccumulation of the inorganic salts. Maintaining in vitro cultures under optimized and controlled conditions produced mycelium with a better composition and health properties than otherwise. Such enriched biomass may be classified as potential functional foods, aiding in overcoming deficiencies of elements and organic compounds with biological activity in humans.

Photostability of Topical Agents Applied to the Skin: A Review.

Topical treatment modalities have multiple advantages starting with the convenient application and non-invasive treatment and ending with the reduction of the risk of the systemic side effects. Active pharmaceutical substances must reach the desired concentration at the target site in order to produce a particular therapeutic effect. In contrast to other dosage forms topical agents applied to the skin may also be susceptible to photodegradation after application. That is why the knowledge of the susceptibility of these topical drugs to UV irradiation, which may contribute to their degradation or changes in chemical structure, is very important. Active pharmaceutical substances used in dermatology may differ both in chemical structure and photostability. Furthermore, various factors-such as light intensity and wavelength, pH, temperature, concentration-can influence the photodegradation process, which is reflected in particular in kinetics of photodegradation of active pharmaceutical substances as well as both the quantitative and qualitative composition of by-products. The aim of this study was to conduct a systematic review of the photostability of dermatological drugs, as well as of other substances commonly applied topically. The photostability of glucocorticosteroids, retinoids, and antifungal drugs as well as non-steroidal anti-inflammatory drugs applied topically and selected UV-filters have been discussed. Furthermore, the impact of photoinstability on the effectiveness of pharmacotherapy and some photostabilization strategies have been also included.

Antidepressant-like activity of hyperforin and changes in BDNF and zinc levels in mice exposed to chronic unpredictable mild stress.

Chronic unpredictable mild stress (CUMS) - a rodent model of depression mimics a variety of neurochemical and behavioral alterations similar to those seen in human depression. This study evaluated the antidepressant activity of hyperforin in the CUMS model using fluoxetine (FLX) as a reference drug. The antidepressant-like effects of hyperforin and FLX were evaluated in the tail suspension test (TST), forced swim test (FST), and splash test (SPT). CUMS induced an increase in immobility time in mice (pro-depressive effects) in the FST and TST. CUMS-induced changes were reversed by chronic treatment with hyperforin (2.5 and 5 mg/kg), as well as FLX (10 mg/kg). SPT results revealed a decrease in the frequency and duration of grooming in stressed mice. These effects were normalized by hyperforin (5 mg/kg) and FLX treatment. Hyperforin (2.5 mg/kg) only reversed the CUMS-induced deficits related to the frequency of grooming. CUMS also caused a decrease in zinc concentration in the frontal cortex (FC) and hippocampus (Hp) of mice; hyperforin (2.5 mg/kg) increased zinc concentration in the Hp of control rats. CUMS also induced a decrease in BDNF protein levels in the FC and Hp, while decreasing the pCREB/CREB ratio only in the Hp. Hyperforin (2.5 and 5 mg/kg) reversed the CUMS-induced reduction of BDNF only in the Hp. Our results demonstrate the antidepressant-like activity of hyperforin in the CUMS model in mice and the possible involvement of hippocampal BDNF/zinc alterations in this activity.

Mycoremediation of azole antifungal agents using in vitro cultures of Lentinula edodes.

Azole antifungal agents are widely used as active ingredients in antifungal pharmaceuticals and agricultural fungicides. An increase in the use of azole antifungals has resulted in an increase in the concentration of these compounds in wastewater and surface water, with potential implications for agriculture. In the present study, bifonazole (BIF) and clotrimazole (CTZ) were selected for investigation because of their widespread use in topical formulations and persistence in the environment. The mycoremediation capacity of BIF and CTZ by mycelia of Lentinula edodes in in vitro culture was evaluated. The main aim of this study was to identify the presumable biodegradation products of the investigated active pharmaceutical substances using the LC/MS/MS method. For this purpose, the media were enriched with the following active pharmaceutical ingredients selected for this study: BIF powder, CTZ powder, and BIF cream, each of them at the same concentration of 0.1 mg/mL. Subsequently, thin-layer chromatography coupled with densitometry was used to evaluate the content of BIF and CTZ in mycelium from in vitro cultures of L. edodes. The degradation process was found to affect primarily the imidazole moiety of both investigated compounds. In addition, the amounts of undegraded investigated compounds were found to be 4.98, 9.26, and 4.56 mg/g dry weight for BIF powder, CTZ powder, and BIF cream, respectively. Therefore, the findings of this study revealed that L. edodes could be considered for remediation of pollution caused by azole antifungal agents.

Assessing the Bioavailability of Zinc and Indole Compounds from Mycelial Cultures of the Bay Mushroom Imleria badia (Agaricomycetes) Using In Vitro Models.

Zinc and indole compounds demonstrate anti-inflammatory, antidepressant, and antioxidant activity. Edible mushrooms are good sources of these substances. Therefore, in this study, we aimed to study the accumulation, release, and absorption of zinc and indole compounds from mycelial cultures of Imleria badia species using in vitro models. Samples were analyzed using the atomic absorption spectroscopy method and the reversed-phase high-performance liquid chromatography method. The highest quantities of zinc were detected in the material grown on zinc hydrogen aspartate-enriched media (176.01 mg/100 g dry weight [d.w.]). In addition, the quantity of zinc in the control biomass was approximately 12.13 mg/100 g d.w. After passive transport, the amount of zinc was detected to be around 1.40 mg/100 g d.w., whereas after active transport with CaCo-2 cells, the quantity of zinc ranged from 0.46 mg/100 g d.w. to 12.72 mg/100 g d.w. Among the organic compounds, four indole compounds were qualitatively identified, including 5-hydroxy-l-tryptophan, melatonin, l-tryptophan, and 5-methyltryptamine. These results indicate that mushrooms and their in vitro cultures not only synthesize and accumulate these compounds, but also potentially release them into the gastrointestinal tract where they can be absorbed by the human body, which is reflected as a specific health benefit.

Lentinula edodes Mycelium as Effective Agent for Piroxicam Mycoremediation.

Pollution of the environment with inorganic and organic substances is one of the main problems in the world. For this reason, it is necessary to conduct researches for effective methods of biodegradation of xenobiotics, including drugs whose unmetabolized forms are introduced into the environment, especially into water. One possible solution to this problem may be the use of white rot fungi, such as Lentinula edodes. This is an edible species used in medicine because of its beneficial anti-cancer, hypocholesterolemic, hypotensive, hypoglycemic and antioxidant effects. Due to the fact that the mycelium of L. edodes produces enzymes with oxidizing properties that can degrade xenobiotics. The aim of the work was verification if in vitro cultures of L. edodes can be used for bioremediation of non-steroidal, anti-inflammatory drug: piroxicam. For this purpose, the in vitro culture of L. edodes was derived and the mycelial cultures of this species enriched with piroxicam were analyzed. The biodegradation pathway of piroxicam by L. edodes mycelium was carried out by the UPLC/MS/MS method. The degradation process of piroxicam was found to affect primarily the linker between the thiazine and the piperidine ring, leading to its oxidation and cleavage. Additionally, oxidation of the benzothiazine moiety was observed, leading to hydroxylation and oxidation of the phenyl ring as well as oxidation of the thiazine ring leading to partial or complete removal of the sulfonamide moiety. It seems that the degradation process led finally to 2-hydroxybenozquinone, which may be further oxidized to inorganic compounds. What's more, concentration of piroxicam in in vitro cultures of L. edodes was not correlated with effectiveness of biodegradation of this compound - on each experimental series, the level of degradation was the same. The results confirm the possibility of using the investigated L. edodes mycelium for remediation of piroxicam.