Search Results (924)

In this study, a new, patented form of a water-soluble cannabidiol (CBD) complex was synthesised and tested. The formation of the complex is based on the interactions, presumably through hydrogen bonding, between cannabidiol and glucosamine, the latter contributing significantly to the increased hydrophilicity. The complex was characterised by chromatography, thermal analysis, nuclear magnetic resonance, Fourier transform infrared spectroscopy, and permeability tests. This complex has a substantially higher water solubility than normal CBD. Permeability tests indicate that it has almost five times lower permeability through lipophilic membranes and less than half the membrane mass retention of conventional CBD. At the same time, its equilibrium concentration is almost four times higher than that of normal CBD. These results suggest that this new form of CBD is a promising candidate for future biological and clinical studies, as it offers improved bioavailability and biodistribution. Full article

The article presents the first method based on high-performance liquid chromatography and ultraviolet detection (HPLC-UV) for the determination of timonacic (thioproline, 1,3-thiazolidine-4-carboxylic acid, tPro) in pharmaceutical tablets and face care products (creams, sera, foundations, suncreams). Sample preparation primarily involves solid-liquid extraction (SLE) of tPro with 0.2 mol/L phosphate buffer pH 6, derivatization with 0.25 mol/L 2-chloro-1-methylquinolinium tetrafluoroborate (CMQT), followed by polytetrafluoroethylene (PTFE) membrane filtration. The chromatographic separation of the stable UV-absorbing 2-S-quinolinium derivative is achieved within 14 min at 25 °C on a Zorbax SB-C18 (150 × 4.6 mm, 5 µm) column using gradient elution. The eluent consists of 0.1 mol/L trichloroacetic acid (TCA), pH 1.7, in a mixture with acetonitrile (ACN) delivered at a flow rate of 1 mL/min. The analyte is quantified by monitoring at 348 nm. The assay linearity was observed within 0.5–125 μmol/L. The limit of quantification (LOQ) was found to be 0.5 μmol/L. The accuracy ranged from 93.22% to 104.31% and 97.38% to 103.48%, while precision varied from 0.30% to 11.23% and 1.13% to 9.64% for intra- and inter-assay measurements, respectively. The method was successfully applied to commercially available on the Polish market pharmaceutical and cosmetic products. Full article

In this study, a magnet-integrated fabric phase sorptive extraction (MI-FPSE) protocol was developed in combination with high pressure liquid chromatography—diode array detection (HPLC-DAD) for the simultaneous determination of five phenylurea pesticides (i.e., chlorbromuron, diuron, linuron, metoxuron, monuron) in environmental water samples. To produce the MI-FPSE device, two individual sol-gel coated carbowax 20 M (CW 20 M) cellulose membranes were fabricated and stitched to each other, while a magnetic rod was inserted between them to give the resulting device the ability to spin and serve as a stand-alone microextraction platform. The adsorption and desorption step of the MI-FPSE protocol was optimized to achieve high extraction efficiency and the MI-FPSE-HPLC-DAD method was validated in terms of linearity, sensitivity, selectivity, accuracy, and precision. The limits of detection (LODs) were found to be 0.3 μg L⁻¹. The relative recoveries were 85.2–110.0% for the intra-day and 87.7–103.2% for the inter-day study. The relative standard deviations were better than 13% in all cases. The green character and the practicality of the developed procedure were assessed using ComplexGAPI and Blue Analytical Grade Index metric tools, showing good method performance. Finally, the developed method was successfully used for the analysis of tap, river, and lake water samples. Full article

Thiophene derivatives are particularly attractive for application in drug development for their versatile pharmacological properties. We synthesized a series of four compounds with thiophene carboxamide as a scaffold. The structures were established based on HR-MS and 1D- and 2D-NMR. The purity of the compounds was established to be greater than 92% by thin-layer chromatography and NMR. The cytotoxic effects of the newly synthesized compounds were evaluated against the normal HaCaT cell line and A375, HT-29, and MCF-7 cancer cell lines. The cytotoxic assessment revealed that two compounds exhibit a significant cytotoxic effect on all cancer cell lines. To investigate their potential underlying mechanisms of action, several tests were performed: immunofluorescence imaging, caspase-3/7 assay, mitochondrial membrane potential (JC-1) assay, and 2′,7′–dichlorofluorescein diacetate (DCFDA) assay. MB-D2 proved to be the most cytotoxic and effective in terms of caspase 3/7 activation, mitochondrial depolarization and decrease in ROS production; these effects did not occur in normal HaCaT cells, revealing that MB-D2 has a high selectivity against A375 cancer cells. Full article

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by restricted social communication and repetitive behaviors. Prenatal stress is critical in neurodevelopment and increases risk for ASD, particularly in those with greater genetic susceptibility to stress. Docosahexaenoic acid (DHA) is one of the most abundant ω-3 fatty acids in the membrane phospholipids of the mammalian brain, and dietary DHA plays an important role in brain development and maintenance of brain structure. In this study, we investigated whether peri-natal supplementation of DHA can alleviate autistic-like behaviors in a genetic risk/stress mouse model and how it alters lipid peroxidation activity and GABAergic system gene expression in the forebrain. Pregnant heterozygous serotonin transporter knockout (SERT-KO) and wild-type (WT) dams were placed in either non-stressed control conditions or chronic variable stress (CVS) conditions and fed either a control diet or a DHA-rich (1% by weight) diet. Offspring of each group were assessed for anxiety and autism-associated behavior at post-natal day 60 using an open field test, elevated plus maze test, repetitive behavior, and the 3-chamber social approach test. A liquid chromatography-mass spectrometry (LC-MS)-based method was used to follow changes in levels of lipid peroxidation products in the cerebral cortex. Male offspring of prenatally stressed SERT-het KO dams exhibited decreased social preference behaviors and increased repetitive grooming behaviors compared to WT control offspring. Moreover, DHA supplementation in male SERT-het mice decreased frequency of grooming behaviors albeit showing no associated effects on social behaviors. Regardless of stress conditions, supplementation of DHA to the WT mice did not result in alterations in grooming nor social interaction in the offspring. Furthermore, no apparent changes were observed in the lipid peroxidation products comparing the stressed and non-stressed brains. Gad2 was downregulated in the cortex of female offspring of prenatally stressed SERT-KO dams, and this change appeared to be rescued by DHA supplementation in offspring. Gad2 was upregulated in the striatum of male offspring of prenatally stressed SERT-KO dams, but DHA did not significantly alter the expression compared to the control diet condition. Full article

Multidrug resistance (MDR) significantly contributes to colon cancer recurrence, making it essential to understand its molecular basis for improved therapies. This study aimed to identify genes and pathways involved in resistance to standard chemotherapeutics by comparing transcriptome profiles of sensitive and paclitaxel-induced MDR colonospheres. Cell viability and growth were assessed following treatment with 5-fluorouracil, oxaliplatin, irinotecan, bevacizumab, and cetuximab. Drug concentrations in culture media posttreatment were measured using high-performance liquid chromatography (HPLC). RNA sequencing (RNA-seq) of untreated sensitive and resistant colonospheres identified differentially expressed genes linked to baseline resistance. Our results confirmed cross-resistance in the resistant model, showing highest oxaliplatin tolerance may involve mechanisms beyond efflux. Transcriptome analysis highlighted upregulation of PIGR and activation of the ribosomal signaling pathway as potential resistance mediators. Notably, AKR1B10, a gene linked to chemotherapeutic detoxification, was overexpressed, whereas genes related to adhesion and membrane transport were downregulated. The overexpression of ribosomal protein genes suggests ribosome biogenesis plays a key role in acquired resistance. These findings suggest that targeting ribosome biogenesis and specific deregulated genes such as PIGR and AKR1B10 may offer promising strategies to overcome MDR in colon cancer. Full article

Fatty acids (FAs) play a crucial role in human physiology, including energy production and serving as signaling molecules. However, a dysregulation in their balance can lead to multiple disorders, such as obesity and metabolic syndrome. These pathological conditions alter the balance between the heart’s energetic substrates, promoting an increased reliance on FAs and decreased cardiac efficiency. A therapeutic application of a non-psychotropic phytocannabinoid, cannabigerol (CBG), seems to be a promising target since it interacts with different receptors and ion channels, including cannabinoid receptors—CB₁ and CB₂, α2 adrenoceptor, or 5-hydroxytryptamine receptor. Therefore, in the current study, we evaluated a concentration-dependent effect of CBG (2.5 µM, 5 µM, and 10 µM) on H9c2 cardiomyocytes in lipid overload conditions. Gas–liquid chromatography and Western blotting techniques were used to determine the cellular lipid content and the level of selected proteins involved in FA metabolism, glucose transport, and the insulin signaling pathway. The glucose uptake assay was performed using a colorimetric method. Eighteen-hour CBG treatment in the highest concentration (10 µM) significantly diminished the accumulation of diacylglycerols (DAGs) and the saturation status of this lipid fraction. Moreover, the same concentration of CBG markedly decreased the level of FA transporters, namely fatty acid translocase (CD36) and plasma membrane fatty acid-binding protein (FABPpm), in the presence of palmitate (PA) in the culture medium. The results of our experiment suggest that CBG can significantly modulate lipid storage and composition in cardiomyocytes, thereby protecting against lipid-induced cellular dysfunction. Full article

The increasing prevalence of antimicrobial resistance (AMR) demands novel strategies, including the use of plant-derived agents. This study investigates the chemical profile and in vitro antimicrobial activity of essential oil from Lavandula angustifolia (LEO), cultivated in Northeastern Bulgaria. Gas chromatography–mass spectrometry (GC-MS) analysis confirmed the presence of a linalool/linalyl acetate chemotype, characteristic of high-quality lavender oil. LEO demonstrated significant inhibitory activity against Escherichia coli ATCC 25922, with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of 0.31% (v/v) and moderate to weak activity against other Gram-positive and fungal strains. Time–kill assays revealed a concentration-dependent bactericidal effect on E. coli. The addition of LEO at subinhibitory concentrations increased the inhibition zones for all antibiotics. In silico analysis identified functional protein clusters potentially modulated by LEO constituents, including targets related to membrane integrity and metabolic regulation. The findings indicate the potential of lavender essential oil as a natural antimicrobial adjuvant; however, additional in vivo and clinical investigations are necessary to validate its therapeutic use. Furthermore, molecular docking analysis revealed a high binding affinity of linalool and linalyl acetate towards the FabI protein of E.coli, suggesting a potential inhibitory mechanism at the molecular level. Full article

The amino acid membrane transporters of grape species take part in metabolic pathways that play crucial roles in nitrogen trafficking and in the synthesis of secondary metabolites. Therefore, identifying these amino acid transporters and defining their functional properties might have further applications in crop improvement and, hence, relevance to human nutrition. The VvCAT2 (Cation Amino acid Transporter) transporter cDNA has been isolated and cloned into a specific plasmid for over-expression in Escherichia coli. The expressed protein, after purification by Ni²⁺-chelating chromatography, has been functionally characterized in an experimental model of proteoliposomes by measuring the uptake of radiolabeled compounds. Arginine was revealed to be the best substrate, confirming the role of CAT2 in nitrogen trafficking in plant cells and within sub-cellular spaces, given its plausible localization in vacuoles. The transporter activity is modulated by pH, osmotic imbalance and ATP. The transport kinetics have been measured. Overall, the obtained data indicate the capacity of VvCAT2 in transporting arginine, making it a possible target for crop improvement with a relevance to human health. Full article

This study provides a comparative analysis of the post-thaw sperm lipidomic profiles of Alpine and Spanish–Creole goat breeds to explore breed-specific differences in fatty acid composition and their implications for sperm function and reproductive efficiency. Lipids were extracted from cryopreserved semen samples of Alpine (n = 7) and Spanish–Creole (n = 4) mature bucks and subsequently analyzed by gas chromatography–mass spectrometry (GC-MS), with 21 fatty acids identified within the two breeds. Eight of these fatty acids, namely 13:0, 16:0, 18:0, 24:0, 14:1, 18:1 (cis-9), 24:1, and 18:2 showed significant differences (p < 0.05). The levels of 16:0, 18:0, 24:0, 18:1 (cis-9), 18:1, and 18:2 were higher in the Alpine breed, whereas the levels of 13:0, 14:1, and 24:1 were higher in the Spanish–Creole breed (p < 0.05). Of those, 16:0, 18:1 (cis-9), and 18:2 were both statistically and biologically significant (p < 0.05, FC > 2). Concentrations of the total fatty acids, total saturated fatty acids (Total-SFA), and total polyunsaturated fatty acids (Total-PUFA) were significantly higher in the Alpine breed, whereas the concentrations of the total cis-monounsaturated fatty acid (Total cis-MUFA) were significantly higher in the Spanish–Creole breed (p < 0.05). Network and pathway analyses revealed that 16:0, 18:1 (cis-9), and 18:2 contributed to the most central nodes of the lipidomic network, which may support membrane stability and cryotolerance. The lipidomic differences observed between breeds may be attributed to both genetic and environmental factors and may provide valuable tools for enhancing breeding strategies, artificial insemination programs, and sperm cryopreservation techniques. Full article

Extracellular vesicles (EVs) are lipid membrane-enclosed particles released by all cells and can be isolated from various sources, even from solid tissues. This study focuses on isolating and characterizing EVs from mouse lymph nodes (LNs). Male C57BL/6 mice were injected with complete Freund’s adjuvant, with or without ovalbumin. Inguinal and popliteal LNs were incised 9 days after immunization, and EV isolation was carried out using a combination of differential centrifugation and size-exclusion chromatography. The characteristic morphology of small and large EVs was confirmed by transmission electron microscopy. Particle size distribution and concentration were determined by nanoparticle tracking analysis, while protein and lipid contents were measured by bicinchoninic acid assay, and sulfo-phospho-vanillin assays, respectively, to calculate the protein-to-lipid ratio. Immune and EV markers were analyzed by using flow cytometry and Western blot assay, revealing significant changes between immunized mice compared to controls. This study establishes a novel protocol for isolating and characterizing EVs from LNs and highlights the impact of immunization on EV properties, offering insights into their roles in immune processes. Full article

Sediments are key players in the optimum functioning of ecosystems; however, they also represent the largest known repository of harmful contaminants. The vast variety of these sediment-associated contaminants may exert harmful effects on marine communities and can impair ecosystem functioning. Whole-cell biosensors are a rapid and biologically relevant tool for assessing environmental toxicity. Therefore, in this study, we developed a bioassay-based toxicity measurement system using genetically modified bacteria to create a whole-cell optical biosensor. Briefly, reporter bacteria were integrated and immobilized using a calcium alginate matrix on fiber-optic tips connected to a photon counter placed inside a light-proof, portable case. The calcium alginate matrix acts as a semi-permeable membrane that protects the reporter-encapsulated optical fiber tips and allows the inward passage of toxicant(s) to induce a dose-dependent response in the bioreporter. The samples were tested by directly submerging the fiber tip with immobilized bacteria into vials containing either water or suspended sediment samples, and the subsequent bioluminescent responses were acquired. In addition to bioavailable sediment toxicity assessments, conventional chemical methods, such as liquid chromatography–mass spectroscopy (LC-MS) and inductively coupled plasma optical emission spectroscopy (ICP-OES), were used for comprehensive evaluation. The results demonstrated the efficacy of the biosensor in detecting various toxicity levels corresponding to identified contaminants, highlighting its potential integration into environmental monitoring frameworks for enhanced sediment and water quality assessments. Despite its utility, this study notes the system’s operational challenges in field conditions, recommending future enhancements for improved portability and usability in remote locations. Full article

Background: The rise in multidrug-resistant bacteria has intensified the search for new antibiotics, drawing attention to essential oils (EOs) for their antimicrobial properties. For this reason, this study focuses on the antimicrobial action of the EO obtained from Tagetes minuta and its impact on bacterial membranes. Methods: The EO was chemically characterized by chromatography–mass spectrometry, and its antimicrobial activity and its effects on surface and bacterial membrane were assessed by using Zeta potential, membrane transition temperature (Tm) determination; and fluorescence spectroscopy with Laurdan and Di-8 ANEPPS. Results: Twenty-seven compounds could be identified, with (E)-Tagetone, (Z)-Ocimenone, and β-pinene as the most abundant. Afterward, the EO was tested against Escherichia coli (MIC and MBC = 17 mg/mL) and Staphylococcus aureus (MIC = 8.5 mg/mL; MBC > 17 mg/mL), showing antimicrobial action in both bacteria, being more effective against E. coli. Mechanistic studies revealed that the EO interacts with bacterial membranes, increasing the Zeta potential by more than 9 mV and enhancing membrane permeability up to 90%. These effects were further confirmed using model lipid membranes, where the EO induced significant changes in membrane properties, including a reduction in dipole potential and transition temperature, suggesting that some EO components could be inserted into the lipid bilayer, disrupting membrane integrity. Conclusions: The EO from T. minuta demonstrates efficient antimicrobial activity by compromising bacterial membrane structure, highlighting its potential as a natural antimicrobial agent. Full article

The growing presence of organic micropollutants (OMPs) in water sources is a major health concern. Successful removal of OMPs from water sources and ensuring the cleanliness of drinking water has become an important topic in recent years. In this study, 15 nanofiltration (NF) and reverse osmosis (RO) commercial membranes were selected and their potential to remove 10 frequently encountered OMPs in drinking water, with systematically different chemical characteristics, was evaluated. To quickly identify the most promising membranes, high throughput dead-end filtrations were initially conducted. Subsequently, the 4 best performing membranes were used in a more relevant high-throughput cross-flow filtration. Membrane performance was evaluated by analyzing OMP concentrations in the feed and retentates of the different membranes using ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS). This study identified NF 90 (Dow), NF 270 (Dow), NFX (Synder) and TS80 (Trisep) as membranes with superior performance, with a permeance between 3 and 7 L.${\mathrm{m}}^{-2}$.h⁻¹.bar⁻¹ and retentions that were generally around 90%, except for NFX which showed slightly lower retentions. Full article

In the present study, the primary by-products of the hemp-seed oil process—hemp seed cake flour and hemp seed protein concentrate—underwent enzymatic hydrolysis using proteases and carbohydrases, either individually or in combination. The effectiveness of these enzymatic treatments in releasing bioactive compounds was evaluated by assessing the antioxidant and anti-inflammatory properties of the aqueous extracts of both hydrolysed and untreated hemp by-products. The aim was to explore their potential senotherapeutic properties and promote their application as dietary supplements. Secondary metabolites such as flavonoids, phenolic acids, and catechins were analysed using high-performance liquid chromatography. Total phenolic, flavonoid, and protein contents were determined using spectrophotometric methods. Scavenging activity (2,2-Diphenyl-1-picrylhydrazyl scavenging assay (DPPH assay)), antioxidant power (Ferric reducing antioxidant power assay (FRAP assay)), and lipid peroxidation-reducing activity (thiobarbituric acid-reactive substance analysis) were assessed through in vitro assays. Possible anti-inflammatory effects were evaluated by assessing haemolysis inhibition. The impact of extracts on albumin glycation induced by exposure to fructose was also determined. To assess the toxicity of extracts, a zebrafish larvae model was employed. All extracts contained significant amounts of phenolic compounds, flavonoids, and proteins, and they exhibited notable activities in reducing lipid peroxidation and stabilising erythrocyte cell membranes. However, they did not significantly influence protein glycation (the glycation inhibition was only in the range of 15–40%). Our research demonstrates the substantial health-promoting potential, including senescence delay, of aqueous extracts from by-products of the hemp-seed oil process, which are available in large quantities and can serve as valuable supplements to support the health of animals, including humans, rather than being discarded as waste from oil production. Full article