Sialyltransferase Mutations Alter the Expression of Calcium-Binding Interneurons in Mice Neocortex, Hippocampus and Striatum.

Gangliosides are major glycans on vertebrate nerve cells, and their metabolic disruption results in congenital disorders with marked cognitive and motor deficits. The sialyltransferase gene St3gal2 is responsible for terminal sialylation of two prominent brain gangliosides in mammals, GD1a and GT1b. In this study, we analyzed the expression of calcium-binding interneurons in primary sensory (somatic, visual, and auditory) and motor areas of the neocortex, hippocampus, and striatum of St3gal2-null mice as well as St3gal3-null and St3gal2/3-double null. Immunohistochemistry with highly specific primary antibodies for GABA, parvalbumin, calretinin, and calbindin were used for interneuron detection. St3gal2-null mice had decreased expression of all three analyzed types of calcium-binding interneurons in all analyzed regions of the neocortex. These results implicate gangliosides GD1a and GT1b in the process of interneuron migration and maturation.

Environmentally Friendly Approach to the Synthesis of 3-[Benzylideneamino]-2-methylquinazolin-4(3H)-one Derivatives and Calculation of Their Toxicity.

Application of deep eutectic solvents in synthesis of different heterocyclic compounds was proven very efficient. These solvents are a new generation of green solvents showing excellent potential for different purposes, where they are used as environmentally acceptable substitute for toxic and volatile organic solvents. This research describes their application in the synthesis of series of quinazolinone Schiff bases in combination with microwave, ultrasound-assisted and mechanochemical methods. First, a model reaction was performed in 20 different deep eutectic solvents to find the best solvent and then reaction conditions (solvent, temperature and reaction time) were optimized for each method. Afterwards, 40 different quinazolinone derivatives were synthesized in choline chloride/malonic acid (1 : 1) DES by each method and compared by their yields. Here we show that deep eutectic solvents can be very efficient in the synthesis of quinazolinone derivatives as an excellent substitution for volatile organic solvents. With green chemistry approach in mind, we have also performed a calculation on compounds' toxicity and solubility, showing that most of them possess toxic and mutagenic properties with low water solubility.

Towards a Greener Approach for Biomass Valorization: Integration of Supercritical Fluid and Deep Eutectic Solvents.

A green and sustainable procedure for obtaining Lavandula stoechas extracts with antioxidant and antimicrobial properties was investigated. Green solvents, supercritical CO2, and natural deep eutectic solvents (NADES) together with ultrasound-assisted extraction were used for the sequential extraction of terpene and polyphenols fractions. After the CO2 extraction of the terpene fraction, the residue material was used in an extraction with different NADES (betaine-ethylene glycol (Bet:EG), betaine-glycerol (Bet:Gly), and glycerol-glucose (Gly:Glu)), intensified with an ultrasound-assisted method (at 30 and 60 °C). In the CO2 extract, the major group of components belonged to oxygenated monoterpenes, while the highest polyphenol content with the dominant rutin (438.93 ± 4.60 µg/mL) was determined in Bet:EG extracts (60 °C). Bet:EG extracts also exhibited the most potent antioxidant activity according to DPPH, ABTS, and FRAP assays. Moreover, Bet:EG extracts showed significant inhibitory activity against Gram-positive and Gram-negative bacteria, with minimum inhibitory activity of 0.781-3.125 and 1.563-6.250 mg·mL-1, respectively. By comparing the polyphenolic content and antioxidant and antimicrobial activities of Bet:EG extracts with extracts obtained with conventional solvents (water and ethanol), the superiority of NADES was determined. The established environmentally friendly procedure unifies the requirements of green and sustainable development and modern pharmacognosy because it combines the use of safe alternative solvents, the absence of solvent waste generation, more rational use of resources, and the attainment of safe and quality extracts.

Terpenes and Cannabinoids in Supercritical CO2 Extracts of Industrial Hemp Inflorescences: Optimization of Extraction, Antiradical and Antibacterial Activity.

Natural products are increasingly in demand in dermatology and cosmetology. In the present study, highly valuable supercritical CO2 (sCO2) extracts rich in bioactive compounds with antiradical and antibacterial activity were obtained from the inflorescences of industrial hemp. Volatile compounds were analyzed by gas chromatography in tandem with mass spectrometry (GC-MS), while cannabinoids were determined by high performance liquid chromatography (HPLC-DAD). Extraction yields varied from 0.75 to 8.83%, depending on the pressure and temperature applied. The extract obtained at 320 bar and 40 °C with the highest content (305.8 µg mg-1) of cannabidiolic acid (CBDA) showed the best antiradical properties. All tested extract concentrations from 10.42 µg mL-1 to 66.03 µg mL-1 possessed inhibitory activities against E. coli, P. aeruginosa, B. subtilis, and S. aureus. The sCO2 extract with the highest content of cannabidiol (CBD) and rich in α-pinene, ß-pinene, ß-myrcene, and limonene was the most effective. The optimal conditions for sCO2 extraction of cannabinoids and volatile terpenes from industrial hemp were determined. The temperature of 60 °C proved to be optimal for all responses studied, while the pressure showed a different effect depending on the compounds targeted. A low pressure of 131.2 bar was optimal for the extraction of monoterpenes, while extracts rich in sesquiterpenes were obtained at 319.7 bar. A high pressure of 284.78 bar was optimal for the extraction of CBD.

Green Synthesis of Thiazolidine-2,4-dione Derivatives and Their Lipoxygenase Inhibition Activity With QSAR and Molecular Docking Studies.

Thiazolidinediones are five-membered, heterocyclic compounds that possess a number of pharmacological activities such as antihyperglycemic, antitumor, antiarthritic, anti-inflammatory, and antimicrobial. Conventional methods for their synthesis are often environmentally unacceptable due to the utilization of various catalysts and organic solvents. In this study, deep eutectic solvents were used in the synthesis of thiazolidinedione derivatives that acted as both solvents and catalysts. Initially, a screening of 20 choline chloride-based deep eutectic solvents for thiazolidinedione synthesis, via Knoevenagel condensation, was performed in order to find the most suitable solvent. Deep eutectic solvent, choline chloride, N-methylurea, was proven to be the best for further synthesis of 19 thiazolidinedione derivatives. Synthesized thiazolidinediones are obtained in yields from 21.49% to 90.90%. The synthesized compounds were tested for the inhibition of lipid peroxidation as well as for the inhibition of soy lipoxygenase enzyme activity. The antioxidant activity of the compounds was also determined by the ABTS and DPPH methods. Compounds showed lipoxygenase inhibition in the range from 7.7% to 76.3%. Quantitative structure-activity relationship model (R 2 = 0.88; Q 2 loo = 0.77; F = 33.69) for the inhibition of soybean lipoxygenase was obtained with descriptors Mor29m, G2u, and MAXDP. The molecular docking confirms experimentally obtained results, finding the binding affinity and interactions with the active sites of soybean LOX-3.

Influence of Red Corn Rich in Anthocyanins on Productive Traits, Blood Metabolic Profile, and Antioxidative Status of Fattening Lambs.

In this study, we aimed to evaluate the effects of different proportions of red corn rich in anthocyanins on the diet of fattening lambs considering their productive traits, blood metabolic profile, and antioxidative status. The research was carried out with 30 Merinolandschaf lambs, 90 days old and weaned. The feed mixture for lambs (n = 10) of the control group contained yellow corn, while in the feed mixture of experimental group I (n = 10), yellow corn was replaced with red corn at 50% (RC50), and in experimental group II (n = 10), yellow corn was 100% replaced with red corn (RC100). An automatic three-part differential haematology analyser was used to determine haematological parameters in whole blood, and biochemical parameters were determined in blood serum using a biochemical analyser. A diet containing red corn did not affect productive traits or the majority of the examined parameters. However, higher blood haemoglobin content, increased aspartate aminotransferase and creatine kinase activity, and decreased glucose and non-esterified fatty acids concentrations were found in the serum of RC100 lambs. These results indicate a positive effect of red corn rich in anthocyanins on the metabolic profile without any changes in the productive traits of lambs.

Selenium and Natural Zeolite Clinoptilolite Supplementation Increases Antioxidative Status and Immune Response in Growing Pigs.

Selenium (Se), an essential trace element for human and animal health, is covalently incorporated into amino acids, acts as a cofactor for antioxidant enzymes, and is involved in the maintenance of the immune system. The main goal of this investigation was to show the effect of Se supplementation, at levels slightly higher than the recommended values, combined with natural zeolite clinoptilolite on Se deposition in tissues (muscle and liver) and on the immune and antioxidative status of supplemented growing pigs. The experiment was carried out during a 98 d period on 60 pigs. Pigs were fed a standard feed mixture based on corn and soybean and were divided into four groups, according to the level of dietary selenium supplementation as follows: C-0.3 mg/kg DM organic Se, E1-0.5 mg/kg DM sodium selenite, E2-0.5 mg/kg DM organic selenium; E3-0.5 mg/kg DM organic Se+0.2% zeolite. Higher (P < 0.05) selenium concentrations were determined in the muscle and liver in growing pigs fed with higher organic Se in combination with zeolite compared to the lower organic Se concentration. Addition of organic Se increased (P < 0.05) Se deposition in muscle and liver compared to the equal amount of inorganic Se (E2 vs. E1). Higher organic Se in combination with natural zeolite addition increases (P < 0.05) proportion of pigs' cluster of differentiation (CD)45+ compared to the same amount of inorganic Se and lower organic Se addition. The proportion of CD45+ and CD4+ lymphocytes was higher (P < 0.05) in E3 group compared to the other groups. Higher (P < 0.05) proportion of CD21+ lymphocytes were measured in the E2 and E3 groups compared with the other groups. The highest (P < 0.01) activity of glutathione peroxidase (GSH-Px) in pig erythrocytes was observed in the E3 group, while higher (P < 0.05) activity of glutathione reductase (GR) was in all experimental groups related to the control one. A dietary addition of 0.5 mg/kg DM of organic Se in combination with zeolite (0.2% DM) has increased (P < 0.05) Se deposition in liver, muscle, and blood, compared to the dietary addition of 0.3 mg/kg DM of the organic Se.

Determination of Suitable Macroporous Resins and Desorbents for Carnosol and Carnosic Acid from Deep Eutectic Solvent Sage (Salvia officinalis) Extract with Assessment of Antiradical and Antibacterial Activity.

In this study, for the first time, the adsorption/desorption characteristics of carnosic acid and carnosol from deep eutectic solvent extract of Salvia officinalis on five macroporous resins (HP20, XAD7HP, XAD16N, HP21, HP2MG) were evaluated. The high adsorption and medium desorption capacities of carnosic acid and carnosol as well as antibacterial and antiradical activity from the extract obtained with choline chloride:lactic acid (1:2) on XAD7HP resin indicated that resin was appropriate. To get the optimal separation process, the influence of factors such as adsorption/desorption time and volume of desorbent was further investigated. The results showed that the extract with high antiradical and antibacterial activity was obtained via adsorption and desorption on XAD7HP resin. The extraction efficiencies of the deep eutectic solvents (DESs) recycled once, twice, and thrice were 97.64% (±0.03%), 93.10% (±0.66%), and 88.94% (±1.15%), respectively, for carnosic acid, and 96.63% (±0.04%), 94.38% (±0.27%), and 91.19% (±0.36%), respectively, for carnosol, relative to the initial solvent efficiency. Based on that, this method is a promising basis for the large-scale preparation of extracts from Salvia officinalis with further application in the pharmaceutical or food industry, especially for maintaining the "green" character of the whole process to obtain the appropriate extract.

Lipoxygenase Inhibition by Plant Extracts.

Lipoxygenases are widespread enzymes that catalyze oxidation of polyunsaturated fatty acids (linoleic, linolenic, and arachidonic acid) to produce hydroperoxides. Lipoxygenase reactions can be desirable, but also lipoxygenases can react in undesirable ways. Most of the products of lipoxygenase reactions are aromatic compounds that can affect food properties, especially during long-term storage. Lipoxygenase action on unsaturated fatty acids could result in off-flavor/off-odor development, causing food spoilage. In addition, lipoxygenases are present in the human body and play an important role in stimulation of inflammatory reactions. Inflammation is linked to many diseases, such as cancer, stroke, and cardiovascular and neurodegenerative diseases. This review summarized recent research on plant families and species that can inhibit lipoxygenase activity.

Lipoxygenase Inhibition Activity of Coumarin Derivatives-QSAR and Molecular Docking Study.

Lipoxygenases (LOXs) are a family of enzymes found in plants, mammals, and microorganisms. In animals and plants, the enzyme has the capability for the peroxidation of unsaturated fatty acids. Although LOXs participate in the plant defense system, the enzyme's metabolites can have numerous negative effects on human health. Therefore, many types of research are searching for compounds that can inhibit LOXs. The best quantitative structure-activity relationship (QSAR) model was obtained using a Genetic Algorithm (GA). Molecular docking was performed with iGEMDOCK. The inhibition of lipoxygenase was in the range of 7.1 to 96.6%, and the inhibition of lipid peroxidation was 7.0-91.0%. Among the synthesized compounds, the strongest inhibitor of soybean LOX-3 (96.6%) was found to be 3-benzoyl-7-(benzyloxy)-2H-chromen-2-one. A lipid peroxidation inhibition of 91.0% was achieved with ethyl 7-methoxy-2-oxo-2H-chromene-3-carboxylate. The docking scores for the soybean LOX-3 and human 5-LOX also indicated that this compound has the best affinity for these LOX enzymes. The best multiple linear QSAR model contains the atom-centered fragment descriptors C-06, RDF035p, and HATS8p. QSAR and molecular docking studies elucidated the structural features important for the enhanced inhibitory activity of the most active compounds, such as the presence of the benzoyl ring at the 3-position of coumarin's core. Compounds with benzoyl substituents are promising candidates as potent lipoxygenase inhibitors.

Effects of Defoliation on Phenolic Concentrations, Antioxidant and Antibacterial Activity of Grape Skin Extracts of the Varieties Blaufränkisch and Merlot (Vitis vinifera L.).

Vitis vinifera L. has been highlighted by its many positive effects on human health, including antibacterial, anti-inflammatory and antioxidant activity due to its rich phytochemical content. Removing six basal leaves has great potential to influence the production of phytochemicals. The purpose of this study was to determine the impact of different terms of defoliation on the antioxidant and antibacterial activity of grape skin extracts of the Blaufränkisch and Merlot varieties. The total phenolic concentration, total and individual anthocyanin concentration, antioxidant activity and antibacterial activity on gram-positive and gram-negative human pathogens have been determined. The study was conducted on three treatments (control treatment without defoliation, defoliation immediately after bloom and defoliation before the start of the véraison phase). The results showed statistically significant enhancement of the total phenolic concentration as well as the antioxidant and antibacterial activity in both studied cultivars. Defoliation just after blooming was the preferable defoliation term in the Merlot variety for achieving the highest total anthocyanin concentration, antioxidant activity and significant increase of antibacterial activity against all four investigated bacteria. Defoliation before the start of the véraison phase was the preferable defoliation term for achieving the highest total anthocyanin concentration in the Blaufränkisch variety. In general, treatment of defoliation immediately after bloom was more beneficial compared with the defoliation before the start of the véraison phase.

Assessment of Total Phenolic Content, In Vitro Antioxidant and Antibacterial Activity of Ruta graveolens L. Extracts Obtained by Choline Chloride Based Natural Deep Eutectic Solvents.

Rue (R. graveolens) has been an extensively studied medicinal plant due to its rich phytochemicals content, such as furanocoumarins and flavonoids. The aim of this study was to determine the effects of varying extraction conditions on the total phenolic content, the antioxidant and antibacterial property of rue leaves crude extracts using deep eutectic solvents with different water content. These extraction conditions include the temperature and the extraction time. The extract obtained at 30 °C, with 20% water and at 90 min, with 13.3 µg mL-1 concentration, was found to possess the highest total phenolic content (38.24 ± 0.11 mg of GAE g-1 of DM) and the highest antioxidant activity (72.53 ± 0.31%). In this study, the same extract showed the best antibacterial efficiency against all the tested strains, especially gram-negative P. aeruginosa.

Extraction of Carnosic Acid and Carnosol from Sage (Salvia officinalis L.) Leaves by Supercritical Fluid Extraction and Their Antioxidant and Antibacterial Activity.

Sage (Salvia officinalis L.) is a good source of antioxidant compounds, carnosic acid and carnosol being the prominent ones. Both are soluble in CO2, and our goal was to investigate the application of supercritical CO2 extraction to obtain sage extracts rich in these compounds. The effect of pressure, temperature, and CO2 flow rate on the carnosic acid and carnosol yield was studied. These variables were optimized by response surface methodology (RSM). The pressure significantly affected carnosol extraction, while the extraction of carnosic acid was affected by the pressure, temperature, and CO2 flow rate. Carnosic acid content varied from 0.29⁻120.0 µg mg-1, and carnosol content from 0.46⁻65.5 µg mg-1. The optimal conditions according to RSM were a pressure of 29.5 MPa, a temperature of 49.1°C, and a CO2 flow rate of 3 kg h-1, and the sage extract yield was calculated to be 6.54%, carnosic acid content 105 µg mg-1, and carnosol content 56.3 µg mg-1. The antioxidant activities of the sage extracts were evaluated by the scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH). Sage extract obtained at 30 MPa and 40°C with 2 kg h-1 CO2 flow rate with a carnosic acid content of 72 µg mg-1 and carnosol content of 55 µg mg-1 exhibited the highest antioxidant activity (80.0 ± 0.68%) amongst the investigated supercritical fluid extracts at 25 µg mL-1 concentration. The antimicrobial properties of extracts were tested on four bacterial strains: Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. The extract with a carnosic acid content of 116 µg mg-1 and a carnosol content of 60.6 µg mg-1 was found to be the most potent agent against B. subtilis.

Synthesis, Characterization, Antibacterial and Antioxidant Potency of NSubstituted- 2-Sulfanylidene-1,3-Thiazolidin-4-one Derivatives and QSAR Study.

BACKGROUND: Rhodanine is known for its potential and important role in the medicinal chemistry since its derivatives exhibit a wide range of pharmacological activities such as antibacterial, antifungal, antidiabetic, antitubercular, anti-HIV, antiparasitic, antioxidant, anticancer, antiproliferative and anthelmintic agents. OBJECTIVES: Since N-substituted rhodanine synthons are rarely commercially available, it is desirable to develop a straightforward synthetic approach for the synthesis of these key building blocks. The objective was to synthesize a series of rhodanine derivatives and to investigate their antimicrobial and antioxidant activity. Also, in order to obtain an insight into their structure-activity relationship, QSAR studies on the antioxidant activity were performed. METHODS: 1H and 13C FTNMR spectra were recorded on Bruker Avance 600 MHz NMR Spectrometer, mass analysis was carried out on ESI+ mode by LC-MS/MS API 2000. 2,2-Diphenyl-1- picrylhydrazyl radical scavenging activity (% DPPH) was determined in dimethylsulfoxide (DMSO) as a solvent. The antibacterial activity was assessed against Bacillus subtilis, Staphylococcus aureus (Gram positive) and Escherichia coli, Pseudomonas aeruginosa (Gram negative) bacteria in terms of the minimum inhibitory concentrations (MICs) by a modified broth microdilution method. RESULTS: A series of N-substituted-2-sulfanylidene-1,3-thiazolidin-4-ones were synthesized and characterized by 1H NMR, 13C NMR, FTIR, GC MS, LCMS/MS and C,H,N,S elemental analysis. Most of the synthesized compounds showed moderate to excellent antibacterial activity (MIC values from 125 µg/ml to 15.62 µg/mL) and DPPH scavenging activity (from 3.60% to 94.40%). Compound 2-thioxo-3- (4-(trifluoromethyl)-phenyl)thiazolidin-4-one showed the most potent activity against Escherichia coli (3.125 µg/mL), equivalent to antibiotic Amikacin sulphate and against Staphylococcus aureus (0.097 µg/ml), 100 times superior then antibiotic Amikacin sulphate. It has also shown a potent antioxidant activity (95% DPPH scavenging). Two best QSAR models, obtained by GETAWAY descriptor R7p+, Balabans molecular connectivity topological index and Narumi harmonic topological index (HNar), suggest that the enhanced antioxidant activity is related to the presence of pairs of atoms higher polarizability at the topological distance 7, substituted benzene ring and longer saturated aliphatic chain in N-substituents. CONCLUSION: A series of novel N-substituted-2-thioxothiazolidin-4-one derivatives were designed, synthesized, characterized and evaluated for their antibacterial and antioxidant activity in vitro. Majority of the compounds showed excellent antibacterial activity compared to ampicillin and few of them have an excellent activity as compared to Chloramphenicol standard antibacterial drug. The QSAR study has clarified the importance of presenting a pairs of atoms higher polarizability, such as Cl and S at the specific distance, as well as the substituted benzene ring and a long saturated aliphatic chain in N-substituents for the enhanced antioxidant activity of 2-sulfanylidene-1,3- thiazolidin-4-one derivatives.

Environmentally Friendly Approach to Knoevenagel Condensation of Rhodanine in Choline Chloride: Urea Deep Eutectic Solvent and QSAR Studies on Their Antioxidant Activity.

A series of rhodanine derivatives was synthesized in the Knoevenagel condensation of rhodanine and different aldehydes using choline chloride:urea (1:2) deep eutectic solvent. This environmentally friendly and catalyst free approach was very effective in the condensation of rhodanine with commercially available aldehydes, as well as the ones synthesized in our laboratory. All rhodanine derivatives were subjected to 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging activity investigation and quantitative structure-activity relationship (QSAR) studies were performed to elucidate their structure-activity relationship. The best multiple linear QSAR model demonstrate a stability in the internal validation and Y-randomization (R² = 0.81; F = 24.225; Q²loo = 0.72; R²Yscr = 0.148). Sphericity of the molecule, ratio of symmetric atoms enhanced atomic mass along the principle axes in regard to total number of atoms in molecule, and 3D distribution of the atoms higher electronegativity (O, N, and S) in molecules are important characteristic for antioxidant ability of rhodanine derivatives. Molecular docking studies were carried out in order to explain in silico antioxidant studies, a specific protein tyrosine kinase (2HCK). The binding interactions of the most active compound have shown strong hydrogen bonding and van der Waals interactions with the target protein.

Design and synthesis of some new 1,3,4-thiadiazines with coumarin moieties and their antioxidative and antifungal activity.

A series of newly disubstituted (compounds 4a,b) and trisubstituted 1,3,4-thiadiazines 5a-l with various substituents was prepared utilizing different thiosemicarbazides and 3-α-bromoacetylcoumarins as starting compounds. The structures of the synthesized 1,3,4-thiadiazines are elucidated and confirmed utilizing the corresponding analytical and spectroscopic data. All of the new thiadiazine derivatives were tested for their antioxidant activity, employing different antioxidant assays (DPPH scavenging activity, iron chelating activity, power reducing activity). Compounds 5b, 5f, 5j and 4b were proven to be the best DPPH radical scavengers, while compounds 5h and 5j have shown the best iron chelating activity. Thiadiazine derivatives were also tested on their antifungal activity against four mycotoxicogenic fungi, Aspergillus flavus, A. ochraceus, Fusarium graminearum and F. verticillioides. The best antifungal against A. flavus was proven to be compound 5e, while compounds 4a and 5c were the best antifungals on A. ochraceus, and compound 5g showed the best antifungal activity on F. verticillioides.