Methylimidazolium ionic liquids - a new class of forever chemicals with endocrine disrupting potential.

A class of chemical with a potentially important perceived future contribution to the net zero carbon goal (as "green" solvents) is the methylimidazolium ionic liquids (MILs). These solvents are used in industrial processes such as biofuel production yet little is known about their environmental stability or toxicity in man although one MIL - 1-octyl-3-methylimidazolium (M8OI) - has been shown to activate the human estrogen receptor alpha (ERα). The stabilities of the chloride unsubstituted methylimidazolium (MI) and MILs possessing increasing alkyl chain lengths (2C, 1-ethyl-3-methylimidazolium (EMI); 4C, 1-butyl-3-methylimidazolium (BMI); 6C; 1-hexyl-3-methylimidazolium (HMI), 8C, M8OI; 10C, 1-decyl-3-methylimidazolium (DMI)) were examined in river water and a human liver model system. The MILs were also screened for their abilities to activate the human ERα in vitro and induce uterine growth in pre-pubertal rats in vivo. Short chain MILs (EMI, BMI and HMI) underwent negligible metabolism and mineralisation in river water; were not metabolised in a model of human liver metabolism; activated the human ERα in vitro and were estrogenic in vivo in rats. A structure-based computational approach predicted short chain MIL binding to both the estrogen binding site and an additional site on the human estrogen receptor alpha. Longer chain MILs (M8OI and DMI) were metabolised in river water and partially mineralised. Based on structure-activity considerations, some of these environmentally-derived metabolites may however, remain a hazard to the population. MILs therefore have the potential to become forever chemicals with adverse effects to both man, other animals and the environment in general.

Mathematical modelling of genipin-bovine serum albumin interaction using fluorescence intensity measurements.

The interaction between genipin and a model protein bovine serum albumin (BSA), with and without the addition of acetic acid, has been studied experimentally and by modelling. The number of amino groups available to react was determined to be 5.6 % of the total number of amino acid building blocks on BSA. Fluorescence intensity was used to record the progress of the reaction over the 24 h, while the modelling study focused on capturing the kinetic profiles of the reaction. The experiments revealed a slow start to the BSA and genipin interaction, that subsequently accelerated in an S-shaped curve which the modelling study linked with the existence of the feedback cycle for both reactive amino groups and genipin. At BSA concentrations ≥30 mg/mL the reaction was accelerated in the presence of acid, while below 30 mg/mL the acidified conditions delayed the onset of the reaction. Contrary to the reaction mechanisms previously proposed, a degree of breakdown of the fluorescent links in the products formed was denoted both experimentally and in a modelling study. This indicated the reversibility of the processes forming fluorescent product/s and suggested feasibility of the successful release of the protein following prospective encapsulation within the genipin-crosslinked hydrogel structure.

Green biosynthesis of bimetallic ZnO@AuNPs with its formulation into cellulose derivative: biological and environmental applications.

Nanoparticles (NPs) formulation in biopolymers is an attractive process for the researcher to decrease the disadvantages of NPs application alone. Bimetallic NPs are a promising formula of two NPs that usually act as synergetic phenomena. Zinc oxide and gold NPs (ZnO@AuNPs) biosynthesis as a bimetallic was prepared via the eco-friendly manner currently. Carboxymethylcellulose (CMC) was employed for the formulation of ZnO@AuNPs as a nanocomposite via a green method. Physicochemical and topographical characterization was assigned to ZnO@AuNPs and nanocomposite features. The nanostructure of bimetallic NPs and nanocomposite were affirmed with sizes around 15 and 25 nm, respectively. Indeed, the DLS measurements affirmed the more reasonable size and stability of the prepared samples as 27 and 93 nm for bimetallic NPs and nanocomposite, respectively. The inhibitory potential of nanocomposite was more than ZnO@AuNPs against Staphylococcus aureus, Escherichia coli, Salmonella typhi, Enterococcus faecalis, Mucor albicans, Aspergillus flavus, and Mucor circinelloid. ZnO@AuNPs and nanocomposite exhibited antioxidant activity via DPPH with IC50 of 71.38 and 32.4 µg/mL, correspondingly. Excellent anti-diabetic potential of nanocomposite with IC50 of 7.4 µg/mL, and ZnO@AuNPs with IC50 of 9.7 µg/mL was reported compared with the standard acarbose with the IC50 of 50.93 µg/mL for amylase inhibition (%). Photocatalytic degradation of RR195 and RB dyes was performed by ZnO@AuNPs and nanocomposite, where maximum degradation was 85.7 ± 1.53 and 88.7 ± 0.58%, respectively using ZnO@AuNPs, 90.3 ± 0.28 and 91.8 ± 0.27%, respectively using nanocomposite at 100 min.

Undifferentiated HepaRG cells show reduced sensitivity to the toxic effects of M8OI through a combination of CYP3A7-mediated oxidation and a reduced reliance on mitochondrial function.

The methylimidazolium ionic liquid M8OI (1-octyl-3-methylimidazolium chloride, also known as [C8mim]Cl) has been detected in the environment and may represent a hazard trigger for the autoimmune liver disease primary biliary cholangitis, based in part on studies using a rat liver progenitor cell. The effect of M8OI on an equivalent human liver progenitor (undifferentiated HepaRG cells; u-HepaRG) was therefore examined. u-HepaRG cells were less sensitive (>20-fold) to the toxic effects of M8OI. The relative insensitivity of u-HepaRG cells to M8OI was in part, associated with a detoxification by monooxygenation via CYP3A7 followed by further oxidation to a carboxylic acid. Expression of CYP3A7 - in contrast to the related adult hepatic CYP3A4 and CYP3A5 forms - was confirmed in u-HepaRG cells. However, blocking M8OI metabolism with ketoconazole only partly sensitized u-HepaRG cells. Despite similar proliferation rates, u-HepaRG cells consumed around 75% less oxygen than B-13 cells, reflective of reduced dependence on mitochondrial activity (Crabtree effect). Replacing glucose with galactose, resulted in an increase in u-HepaRG cell sensitivity to M8OI, near similar to that seen in B-13 cells. u-HepaRG cells therefore show reduced sensitivity to the toxic effects of M8OI through a combination of metabolic detoxification and their reduced reliance on mitochondrial function.

Vac-and-fill: A micromoulding technique for fabricating microneedle arrays with vacuum-activated, hands-free mould-filling.

We report a simple and reproducible micromoulding technique that dynamically fills microneedle moulds with a liquid formulation, using a plastic syringe, triggered by the application of vacuum ('vac-and-fill'). As pressure around the syringe drops, air inside the syringe pushes the plunger to uncover an opening in the syringe and fill the microneedle mould without manual intervention, therefore removing inter-operator variability. The technique was validated by monitoring the plunger movement and pressure at which the mould would be filled over 10 vacuum cycles for various liquid formulation of varying viscosity (water, glycerol, 20 % polyvinylpyrrolidone (PVP) solution or 40 % PVP solution). Additionally, the impact of re-using the disposable syringes on plunger movement, and thus the fill pressure, was investigated using a 20 % PVP solution. The fill pressure was consistent at 300-450 mbar. It produced well-formed and mechanically robust PVP, poly(methylvinylether/maleic anhydride) and hydroxyethylcellulose microneedles from liquid formulations. This simple and inexpensive technique of micromoulding eliminated the air entrapment and bubble formation, which prevent reproducible microneedle formation, in the resultant microneedle arrays. It provides a cost-effective alternative to the conventional micromoulding techniques, where the application of vacuum ('fill-and-vac') or centrifugation following mould-filling may be unsuitable, ineffective or have poor reproducibility.

Antimicrobial, Antidiabetic, Antioxidant, and Anticoagulant Activities of Cupressus sempervirens In Vitro and In Silico.

In the last decade, the urgent need to explore medicinal plants or drug development has increased enormously around the world to overcome numerous health problems. In the present investigation, HPLC indicated the existence of 18 phenolic and flavonoid compounds in the Cupressus sempervirens extract. Hesperetin represents the greatest concentration (25,579.57 µg/mL), while other compounds, such as pyro catechol, rutin, gallic acid, chlorogenic acid, naringenin, and quercetin, were recognized in concentrations of 2922.53 µg/mL, 1313.26 µg/mL, 1107.26 µg/mL, 389.09 µg/mL, 156.53 µg/mL, and 97.56 µg/mL, respectively. The well diffusion method documented the antibacterial/antifungal activity of C. sempervirens extract against E. faecalis, E. coli, C. albicans, S. typhi, S.aureus, and M. circinelloid with 35, 33, 32, 25, 23, and 21 mm inhibition zones, respectively, more than the standard antibiotic/antifungal agent. Low values ranging from 7.80 to 15.62 µg/mL of MIC and MBC were recorded for E. faecalis, E. coli, and C. albicans. From the 1- diphenyl-2-picryl hydrazyl (DPPH) assay, promising antioxidant activity was recorded for C. sempervirens extract with IC50 of an 8.97 µg/mL. Moreover, ferric reducing antioxidant power (FRAP) and total antioxidant capacity assays (TAC) confirmed the antioxidant activity of the extract, which was expressed as the ascorbic acid equivalent (AAE) of 366.9 ± 0.2 µg/mg and 102 ± 0.2 µg/mg of extracts, respectively. α-amylase and α-glucosidase inhibition % were determined to express the antidiabetic activity of the extract in vitro, with promising IC50 value (27.01 µg/mL) for α-amylase compared to that of acarbose (50.93 µg/mL), while IC50 value of the extract for α-glucosidase was 19.21µg/mL compared to that of acarbose 4.13 µg/mL. Prothrombin time (PT) and activated partial thromboplastin time (APTT) revealed the role of C. sempervirens extract as an anticoagulant agent if compared with the activity of heparin. Binding interactions of hesperetin and gallic acid were examined via the Molecular Operating Environment (MOE) Dock software against E. faecalis (PDB ID: 3CLQ), C. albicans (PDB ID: 7RJC), α-amylase (PDB ID: 4W93), and α-glucosidase (PDB ID: 3TOP). The obtained results shed light on how molecular modeling methods might inhibit the tested compounds, which have the potential to be useful in the treatment of target proteins.

Anti-Yeasts, Antioxidant and Healing Properties of Henna Pre-Treated by Moist Heat and Molecular Docking of Its Major Constituents, Chlorogenic and Ellagic Acids, with Candida albicans and Geotrichum candidum Proteins.

Lawsonia inermis, known as henna, has traditionally been utilized in cosmetics and folk medicine because of their valuable health effects. A lack of information about the processes that increase or decrease release, as well as the biological activities of constituents of natural origin, is an important pharmacological problem. This investigation evaluates the influence of moist heat on the flavonoid and phenolic contents of henna powder and their biological activities. HPLC analysis reflected the existence of 20 and 19 compounds of flavonoids and phenolics in the extract of unpre-treated henna by moist heat (UPMH) and pre-treated henna by moist heat (PMH). Several compounds such as chlorogenic acid, ellagic acid, rutin, rosmarinic acid, kaempferol, and pyrocatechol occurred with high concentrations of 57,017.33, 25,821.09, 15,059.88, 6345.08, 1248.42, and 819.19 µg/mL UPMH while occurred with low concentrations of 44,286.51, 17,914.26, 3809.85, 5760.05, 49.01, and 0.0 µg/mL, respectively in PMH. C. albicans, C. tropicalis, and G. candidum were more affected by UPMH with inhibition zones of 30.17 ± 0.29, 27 ± 0.5, and 29 ± 1.5 mm than PMH with inhibition zones of 29 ± 0.5, 25.33 ± 0.58, and 24.17 ± 0.29 mm, respectively. UPMH henna exhibited less MIC and MFC against the tested yeasts than PMH. Moreover, UPMH henna showed good wound healing, where the rat of migration, wound closure %, and area difference % were 14.806 um, 74.938 um2, and 710.667% compared with PMH henna 11.360 um, 59.083 um2, 545.333%, respectively. Antioxidant activity of UPMH and PMH henna. Promising antioxidant activity was recorded for both UPMH or PMH henna with IC50 5.46 µg/mL and 7.46 µg/mL, respectively. The docking interaction of chlorogenic acid and ellagic acid with the crystal structures of G. candidum (4ZZT) and C. albicans (4YDE) was examined. The biological screening demonstrated that the compounds had favorable docking results with particular proteins. Chlorogenic acid had robust behavior in the G. candidum (4ZZT) active pocket and displayed a docking score of -7.84379 Kcal/mol, higher than ellagic acid's -6.18615 Kcal/mol.

Anti-Helicobacter pylori, Antioxidant, Antidiabetic, and Anti-Alzheimer's Activities of Laurel Leaf Extract Treated by Moist Heat and Molecular Docking of Its Flavonoid Constituent, Naringenin, against Acetylcholinesterase and Butyrylcholinesterase.

It is worth noting that laurel (Laurus nobilis L.) contains several pharmacologically and nutritionally active compounds that may differ according to the pretreatment process. The current study is designed to clarify the effect of moist heat on the phenolic and flavonoid constituents and anti-Helicobacter pylori, antioxidant, antidiabetic, and anti-Alzheimer's activities of laurel leaf extract (LLE). Unmoist-heated (UMH) and moist-heated (MH) LLEs showed the presence of numerous flavonoid and phenolic constituents, although at different levels of concentration. MH significantly induced (p < 0.05) the occurrence of most compounds at high concentrations of 5655.89 µg/mL, 3967.65 µg/mL, 224.80 µg/mL, 887.83 µg/mL, 2979.14 µg/mL, 203.02 µg/mL, 284.65 µg/mL, 1893.66 µg/mL, and 187.88 µg/mL, unlike the detection at low concentrations of 3461.19 µg/mL, 196.96 µg/mL, 664.12 µg/mL, 2835.09 µg/mL, 153.26 µg/mL, 254.43 µg/mL, 1605.00 µg/mL, 4486.02 µg/mL, and 195.60 µg/mL using UMH, for naringenin, methyl gallate, caffeic acid, rutin, ellagic acid, coumaric acid, vanillin, ferulic acid, and hesperetin, respectively. Chlorogenic acid, syringic acid, and daidzein were detected in the UMH LLE but not in the MH LLE, unlike pyrocatechol. The anti-H. pylori activity of the UMH LLE was lower (23.67 ± 0.58 mm of inhibition zone) than that of the MH LLE (26.00 ± 0.0 mm of inhibition zone). Moreover, the values of MIC and MBC associated with the MH LLE were very low compared to those of the UMH LLE. Via MBC/MIC index calculation, the UMH and MH LLEs showed cidal activity. The MH LLE exhibited higher anti-biofilm activity (93.73%) compared to the anti-biofilm activity (87.75%) of the MH LLE against H. pylori. The urease inhibition percentage was more affected in the UMH LLE compared to the MH LLE, with significant (p < 0.05) IC50 values of 34.17 µg/mL and 91.11 µg/mL, respectively. Promising antioxidant activity was documented with a very low value of IC50 (3.45 µg/mL) for the MH LLE compared to the IC50 value of 4.69 µg/mL for the UMH LLE and the IC50 value of 4.43 µg/mL for ascorbic acid. The MH LLE showed significantly higher (p < 0.05) inhibition of α-glucosidase and butyrylcholinesterase activities, with IC50 values of 9.9 µg/mL and 17.3 µg/mL, respectively, compared to those of the UMH LLE at 18.36 µg/mL and 28.92 µg/mL. The molecular docking of naringenin showed good docking scores against acetylcholinesterase 1E66 and butyrylcholinesterase 6EMI, indicating that naringenin is an intriguing candidate for additional research as a possible medication for Alzheimer's disease.

Potential for cardiac toxicity with methylimidazolium ionic liquids.

Methylimidazolium ionic liquids (MILs) are solvent chemicals used in industry. Recent work suggests that MILs are beginning to contaminate the environment and lead to exposure in the general population. In this study, the potential for MILs to cause cardiac toxicity has been examined. The effects of 5 chloride MIL salts possessing increasing alkyl chain lengths (2 C, EMI; 4 C, BMI; 6 C; HMI, 8 C, M8OI; 10 C, DMI) on rat neonatal cardiomyocyte beat rate, beat amplitude and cell survival were initially examined. Increasing alkyl chain length resulted in increasing adverse effects, with effects seen at 10-5 M at all endpoints with M8OI and DMI, the lowest concentration tested. A limited sub-acute toxicity study in rats identified potential cardiotoxic effects with longer chain MILs (HMI, M8OI and DMI) based on clinical chemistry. A 5 month oral/drinking water study with these MILs confirmed cardiotoxicity based on histopathology and clinical chemistry endpoints. Since previous studies in mice did not identify the heart as a target organ, the likely cause of the species difference was investigated. qRT-PCR and Western blotting identified a marked higher expression of p-glycoprotein-3 (also known as ABCB4 or MDR2) and the breast cancer related protein transporter BCRP (also known as ABCG2) in mouse, compared to rat heart. Addition of the BCRP inhibitor Ko143 - but not the p-glycoproteins inhibitor cyclosporin A - increased mouse cardiomyocyte HL-1 cell sensitivity to longer chain MILs to a limited extent. MILs therefore have a potential for cardiotoxicity in rats. Mice may be less sensitive to cardiotoxicity from MILs due in part, to increased excretion via higher levels of cardiac BCRP expression and/or function. MILs alone, therefore may represent a hazard in man in the future, particularly if use levels increase. The impact that MILs exposure has on sensitivity to cardiotoxic drugs, heart disease and other chronic diseases is unknown.

Evaluation of Biomedical Applications for Linseed Extract: Antimicrobial, Antioxidant, Anti-Diabetic, and Anti-Inflammatory Activities In Vitro.

BACKGROUND: In the last few decades, the development of multidrug-resistant (MDR) microbes has accelerated alarmingly and resulted in significant health issues. Morbidity and mortality have increased along with the prevalence of infections caused by MDR bacteria, making the need to solve these problems an urgent and unmet challenge. Therefore, the current investigation aimed to evaluate the activity of linseed extract against Methicillin-resistant Staphylococcus aureus (MRSA) as an isolate from diabetic foot infection. In addition, antioxidant and anti-inflammatory biological activities of linseed extract were evaluated. RESULT: HPLC analysis indicated the presence of 1932.20 µg/mL, 284.31 µg/mL, 155.10 µg/mL, and 120.86 µg/mL of chlorogenic acid, methyl gallate, gallic acid, and ellagic acid, respectively, in the linseed extract. Rutin, caffeic acid, coumaric acid, and vanillin were also detected in the extract of linseed. Linseed extract inhibited MRSA (35.67 mm inhibition zone) compared to the inhibition zone (29.33 mm) caused by ciprofloxacin. Standards of chlorogenic acid, ellagic acid, methyl gallate, rutin, gallic acid, caffeic acid, catechin, and coumaric acid compounds reflected different inhibition zones against MRSA when tested individually, but less than the inhibitory action of crude extract. A lower MIC value, of 15.41 µg/mL, was observed using linseed extract than the MIC 31.17 µg/mL of the ciprofloxacin. The MBC/MIC index indicated the bactericidal properties of linseed extract. The inhibition % of MRSA biofilm was 83.98, 90.80, and 95.58%, using 25%, 50%, and 75%, respectively, of the MBC of linseed extract. A promising antioxidant activity of linseed extract was recorded, with an IC50 value of 20.8 µg/mL. Anti-diabetic activity of linseed extract, expressed by glucosidase inhibition, showed an IC50 of 177.75 µg/mL. Anti-hemolysis activity of linseed extract was documented at 90.1, 91.5, and 93.7% at 600, 800, and 1000 µg/mL, respectively. Anti-hemolysis activity of the chemical drug indomethacin, on the other hand, was measured at 94.6, 96.2, and 98.6% at 600, 800, and 1000 µg/mL, respectively. The interaction of the main detected compound in linseed extract (chlorogenic acid) with the crystal structure of the 4G6D protein of S. aureus was investigated via the molecular docking (MD) mode to determine the greatest binding approach that interacted most energetically with the binding locations. MD showed that chlorogenic acid was an appropriate inhibitor for S. aureus via inhibition of its 4HI0 protein. The MD interaction resulted in a low energy score (-6.26841 Kcal/mol) with specified residues (PRO 38, LEU 3, LYS 195, and LYS 2), indicating its essential role in the repression of S. aureus growth. CONCLUSION: Altogether, these findings clearly revealed the great potential of the in vitro biological activity of linseed extract as a safe source for combatting multidrug-resistant S. aureus. In addition, linseed extract provides health-promoting antioxidant, anti-diabetic, and anti-inflammatory phytoconstituents. Clinical reports are required to authenticate the role of linseed extract in the treatment of a variety of ailments and prevent the development of complications associated with diabetes mellitus, particularly type 2.

Anticandidal activity of nanocomposite based on nanochitosan, nanostarch and mycosynthesized copper oxide nanoparticles against multidrug-resistant Candida.

Recently, antimicrobial resistance has increased globally particularly Candida infections. Most of antifungal drugs used for treating candidiasis became resistant to most of Candida species. In the current study, a nanocomposite based on mycosynthesized copper oxide nanoparticles (CuONPs), nanostarch, nanochitosan was prepared. Results illustrated that twenty-four Candida isolates were isolated from clinical samples. Furthermore, three Candida strains were selected as the most resistant among others toward commercial antifungal drugs; these selected strains were identified genetically as C. glabrata MTMA 19, C. glabrata MTMA 21 and C. tropicalis MTMA 24. Characterization of the prepared nanocomposite was carried out using physiochemical analysis included Ultraviolet-visible spectroscopy (Uv-Vis), Fourier-Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), Energy-Dispersive X-ray spectroscopy (EDX) and Transmission Electron Microscopy (TEM). Moreover, the nanocomposite exhibited promising anticandidal activity against C. glabrata MTMA 19, C. glabrata MTMA 21 and C. tropicalis MTMA 24, where the inhibition zones were 15.3, 27 and 28 mm, respectively. Ultrastructure changes observed in nanocomposite-treated C. tropicalis demonstrated disruption of the cell wall which led to cell death. In conclusion, our results confirmed that the novel biosynthesized nanocomposite based on mycosynthesized CuONPs, nanostarch and nanochitosan is a promising anticandidal agent to fight multidrug-resistant Candida.

Pharmacological Evaluation of Acacia nilotica Flower Extract against Helicobacter pylori and Human Hepatocellular Carcinoma In Vitro and In Silico.

The resistance of cancer and Helicobacter pylori to several drugs reflects a worldwide problem, and it has been the intention of numerous researchers to overcome this problem. Thus, in this study, Acacia nilotica fruits were subjected to HPLC analysis to detect their phenolic compounds and flavonoids. Moreover, A. nilotica's anti-H. pylori activity and its inhibitory activity against human hepatocellular carcinoma (HepG-2 cells) were reported. Various compounds with different concentrations, such as ferulic acid (5451.04 µg/mL), chlorogenic acid (4572.26 µg/mL), quercetin (3733.37 µg/mL), rutin (2393.13 µg/mL), gallic acid (2116.77 µg/mL), cinnamic acid (69.72 µg/mL), hesperetin (121.39 µg/mL) and methyl gallate (140.45 µg/mL), were detected. Strong anti-H. pylori activity at 31 mm was reported, compared to the positive control of the 21.67 mm inhibition zone. Moreover, the MIC and MBC were 7.8 µg/mL and 15.62 µg/mL, respectively, while the MIC and MBC of the positive control were 31.25 µg/mL. The concentration of MBC at 25%, 50% and 75% reflected H. pylori's anti-biofilm activity of 70.38%, 82.29% and 94.22%, respectively. Good antioxidant properties of the A. nilotica flower extract were documented at 15.63, 62.50, 250 and 1000 µg/mL, causing the DPPH scavenging percentages of 42.3%, 52.6%, 65.5% and 80.6%, respectively, with a IC50 of 36.74 µg/mL. HepG-2 cell proliferation was inhibited (91.26%) using 500 µg/mL of flower extract with an IC50 of 176.15 µg/mL, compared to an IC50 of 395.30 µg/mL used against human normal melanocytes. Molecular docking was applied to investigate ferulic acid with the H. pylori (4HI0) crystal structure to determine the best binding mode that interacted most energetically with the binding sites. Molecular docking indicated that ferulic acid was a proper inhibitor for the 4HI0 protein enzyme of H. pylori. A low energy score (-5.58 Kcal/mol) was recorded as a result of the interaction of ferulic acid with the residue's SER 139 active site caused by the O 29 atom, which was important for its antibacterial activity.

Phytochemical Characterization and Efficacy of Artemisia judaica Extract Loaded Chitosan Nanoparticles as Inhibitors of Cancer Proliferation and Microbial Growth.

Despite the advanced development in the field of drug discovery and design, fighting infectious and non-infectious diseases remains a major worldwide heath challenge due to the limited activity of currently used drugs. Nevertheless, in recent years, the approach of designing nanoparticles for therapeutic applications has gained more interest and promise for future use. Thus, the current study is focused on the evaluation of A. judaica extract and chitosan nanoparticles loaded extract (CNPsLE) for potential antimicrobial and anticancer activities. The HPLC analysis of the extract has shown the presence of various phenolic and flavonoid compounds, including kaempferol (3916.34 µg/mL), apigenin (3794.32 µg/mL), chlorogenic acid (1089.58 µg/mL), quercetin (714.97 µg/mL), vanillin (691.55 µg/mL), naringenin (202.14 µg/mL), and rutin (55.64 µg/mL). The extract alone showed higher MIC values against B. subtilis, E. coli, S. aureus, K. pneumonia, and C. albicans (62.5, 15.65, 15.62, 31.25, and 31.25 µg/mL, respectively), whereas lower MIC values were observed when the extract was combined with CNPsLE (0.97, 1.95, 3.9, 4.1, and 15.62 µg/mL, respectively). The extract exhibited low cytotoxicity against normal Vero cells with IC50 173.74 µg/mL in comparison with the cytotoxicity of the CNPsLE (IC50, 73.89 µg/mL). However, CNPsLE showed more selective toxicity against the human prostate cancer cell line (PC3) with IC50 of 20.8 µg/mL than the extract alone with 76.09 µg/mL. In the docking experiments, kaempferol and apigenin were revealed to be suitable inhibitors for prostate cancer (2Q7L). Overall, the obtained data highlighted the promising potential therapeutic use of CNPsLE as an anticancer and antimicrobial agent.

The methylimidazolium ionic liquid M8OI is a substrate for OCT1 and p-glycoprotein-1 in rat.

The methylimidazolium ionic liquid M8OI was recently found to be present in both the environment and man. In this study, M8OI disposition and toxicity were examined in an established rat progenitor-hepatocyte model. The progenitor B-13 cell was approx. 13 fold more sensitive to the toxic effects of M8OI than the hepatocyte B-13/H cell. However, this difference in sensitivity was not associated with a difference in metabolic capacities. M8OI toxicity was significantly decreased in a dose-dependent manner by co-addition of the OCT1 (SLC22A1) inhibitor clonidine, but not by OCT2 or OCT3 inhibitors in B-13 cells. M8OI toxicity was also dose-dependently increased by the co-addition of p-glycoprotein-1 (ABCB1B, multi drug resistant protein 1 (MDR1)) substrates/inhibitors. Excretion of B-13-loaded fluorophore Hoechst 33342 was also inhibited by the p-glycoproteins substrate cyclosporin A and by M8OI in a dose-dependent manner. Comparing levels of OCT and p-glycoprotein transcripts and proteins in B-13 and B-13/H cells suggest that the lower sensitivity to M8OI in B-13/H cells is predominantly associated with their higher expression of p-glycoprotein-1. These data together therefore suggest that a determinant in M8OI toxicity in rats is the expression and activity of the p-glycoprotein-1 transporter.

Apigenin attenuates LPS-induced neurotoxicity and cognitive impairment in mice via promoting mitochondrial fusion/mitophagy: role of SIRT3/PINK1/Parkin pathway.

RATIONALE: Alteration of the NAD+ metabolic pathway is proposed to be implicated in lipopolysaccharide (LPS)-induced neurotoxicity and mitochondrial dysfunction in neurodegenerative diseases. Apigenin, a naturally-occurring flavonoid, has been reported to maintain NAD+ levels and to preserve various metabolic functions. OBJECTIVES: This study aimed to explore the effect of apigenin on mitochondrial SIRT3 activity as a mediator through which it could modulate mitochondrial quality control and to protect against intracerebrovascular ICV/LPS-induced neurotoxicity. METHODS: Mice received apigenin (40 mg/kg; p.o) for 7 consecutive days. One hour after the last dose, LPS (12 µg/kg, icv) was administered. RESULTS: Apigenin robustly guarded against neuronal degenerative changes and maintained a normal count of intact neurons in mice hippocampi. Consequently, it inhibited the deleterious effect of LPS on cognitive functions. Apigenin was effective in preserving the NAD+/NADH ratio to boost mitochondrial sirtuin-3 (SIRT3), activity, and ATP production. It conserved normal mitochondrial features via induction of the master regulator of mitochondrial biogenesis, peroxisome proliferator-activated receptor γ (PPARγ) coactivator-1α (PGC-1α), along with mitochondrial transcription factor A (TFAM) and the fusion proteins, mitofusin 2 (MFN2), and optic atrophy-1 (OPA1). Furthermore, it increased phosphatase and tensin homolog (PTEN)-induced putative kinase 1 (PINK1) and parkin expression as well as the microtubule-associated protein 1 light chain 3 II/I ratio (LC3II/I) to induce degradation of unhealthy mitochondria via mitophagy. CONCLUSIONS: These observations reveal the marked neuroprotective potential of apigenin against LPS-induced neurotoxicity through inhibition of NAD+ depletion and activation of SIRT3 to maintain adequate mitochondrial homeostasis and function.

Screening of Bioactive Compounds from Endophytic Marine-Derived Fungi in Saudi Arabia: Antimicrobial and Anticancer Potential.

Nowadays, endophytic fungi represent a rich source of biological active compounds. In the current study, twelve endophytic fungal species were isolated from Avicennia marina leaves. From the isolates, Aspergillus niger, Penicillium rubens and Alternaria alternata recorded the highest isolation frequency (80%), relative density (12.5%) and antimicrobial activity. The antimicrobial and anticancer activities of P. rubens were more effective than those of A. niger and A. alternata; therefore, its identification was confirmed via the ITS rRNA gene. Filtrate extracts of P. rubens, A. alternata and A. niger were analyzed using GC-MS and showed different detected constituents, such as acetic acid ethyl ester, N-(4,6-Dimethyl-2-pyrimidinyl)-4-(4-nitrobenzylideneamino) benzenesulfonamide, 1,2-benzenedicarboxylic acid, hexadecanoic acid and octadecanoic acid. Filtrate extract of P. rubens exhibited the presence of more compounds than A. alternata and A. niger. Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus fumigatus were more inhibited by P. rubens extract than A. alternata or A. niger, with inhibition zones of 27.2 mm, 22.21 mm, 26.26 mm, 27.33 mm, 28.25 mm and 8.5 mm, respectively. We observed negligible cytotoxicity of P. rubens extract against normal cells of human lung fibroblasts (WI-38 cell line), unlike A. alternata and A. niger extracts. Proliferation of prostate cancer (PC-3) was inhibited using P. rubens extract, exhibiting mortality levels of 75.91% and 76.2% at 200 µg/mL and 400 µg/mL of the extract. Molecular docking studies against the crystal structures of C. albicans (6TZ6) and the cryo-EM structure of B. subtilis (7CKQ) showed significant interactions with benzenedicarboxylic acid and N-(4,6-dimethyl-2-pyrimidinyl)-4-(4-nitrobenzylideneamino) benzenesulfonamide as a constituent of P. rubens extract. N-(4,6-dimethyl-2-pyrimidinyl)-4-(4-nitrobenzylideneamino) benzenesulfonamide had the highest scores of −6.04905 kcal/mol and −6.590 kcal/mol towards (6tz6) and (7CKQ), respectively.

Molecular Interaction Studies and Phytochemical Characterization of Mentha pulegium L. Constituents with Multiple Biological Utilities as Antioxidant, Antimicrobial, Anticancer and Anti-Hemolytic Agents.

Multiple biological functions of Mentha pulegium extract were evaluated in the current work. Phytochemical components of the M. pulegium extract were detected by Gas Chromatography-Mass Spectrometry (GC-MS) and High-performance liquid chromatography (HPLC). Moreover, M. pulegium extract was estimated for antioxidant potential by 2,2-Diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free radical scavenging, antimicrobial activity by well diffusion, and anticoagulant activity via prothrombin time (PT) and activated partial thromboplastin time (APTT). GC-MS analysis detected compounds including cholesterol margarate, stigmast-5-en-3-ol, 19-nor-4-androstenediol, androstan-17-one, pulegone-1,2-epoxide, isochiapin B, dotriacontane, hexadecanoic acid and neophytadiene. Chrysoeriol (15.36 µg/mL) was followed by kaempferol (11.14 µg/mL) and 7-OH flavone (10.14 µg/mL), catechin (4.11 µg/mL), hisperdin (3.05 µg/mL), and luteolin (2.36 µg/mL) were detected by HPLC as flavonoids, in addition to ferulic (13.19 µg/mL), cinnamic (12.69 µg/mL), caffeic (11.45 µg/mL), pyrogallol (9.36 µg/mL), p-coumaric (5.06 µg/mL) and salicylic (4.17 µg/mL) as phenolics. Antioxidant activity was detected with IC50 18 µg/mL, hemolysis inhibition was recorded as 79.8% at 1000 µg/mL, and PT and APTT were at 21.5 s and 49.5 s, respectively, at 50 µg/mL of M. pulegium extract. The acute toxicity of M. pulegium extract was recorded against PC3 (IC50 97.99 µg/mL) and MCF7 (IC50 80.21 µg/mL). Antimicrobial activity of M. pulegium extract was documented against Bacillus subtilis, Escherichia coli, Pseudomonasaureus, Candida albicans, Pseudomonas aeruginosa, but not against black fungus Mucor circinelloides. Molecular docking was applied using MOE (Molecular Operating Environment) to explain the biological activity of neophytadiene, luteolin, chrysoeriol and kaempferol. These compounds could be suitable for the development of novel pharmacological agents for treatment of cancer and bacterial infections.

Molecular Docking and Efficacy of Aloe vera Gel Based on Chitosan Nanoparticles against Helicobacter pylori and Its Antioxidant and Anti-Inflammatory Activities.

The medicinal administration of Aloe vera gel has become promising in pharmaceutical and cosmetic applications particularly with the development of the nanotechnology concept. Nowadays, effective H. pylori treatment is a global problem; therefore, the development of natural products with nanopolymers such as chitosan nanoparticles (CSNPs) could represent a novel strategy for the treatment of gastric infection of H. pylori. HPLC analysis of A. vera gel indicated the presence of chlorogenic acid as the main constituent (1637.09 µg/mL) with other compounds pyrocatechol (1637.09 µg/mL), catechin (1552.92 µg/mL), naringenin (528.78 µg/mL), rutin (194.39 µg/mL), quercetin (295.25 µg/mL), and cinnamic acid (37.50 µg/mL). CSNPs and A. vera gel incorporated with CSNPs were examined via TEM, indicating mean sizes of 83.46 nm and 36.54 nm, respectively. FTIR spectra showed various and different functional groups in CSNPs, A. vera gel, and A. vera gel incorporated with CSNPs. Two strains of H. pylori were inhibited using A. vera gel with inhibition zones of 16 and 16.5 mm, while A. vera gel incorporated with CSNPs exhibited the highest inhibition zones of 28 and 30 nm with resistant and sensitive strains, respectively. The minimal inhibitory concentration (MIC) was 15.62 and 3.9 µg/mL, while the minimal bactericidal concentration (MBC) was 15.60 and 7.8 µg/mL with MBC/MIC 1 and 2 indexes using A. vera gel and A. vera gel incorporated with CSNPs, respectively, against the resistance strain. DPPH Scavenging (%) of the antioxidant activity exhibited an IC50 of 138.82 µg/mL using A.vera gel extract, and 81.7 µg/mL when A.vera gel was incorporated with CSNPs. A.vera gel incorporated with CSNPs enhanced the hemolysis inhibition (%) compared to using A.vera gel alone. Molecular docking studies through the interaction of chlorogenic acid and pyrocatechol as the main components of A. vera gel and CSNPs with the crystal structure of the H. pylori (4HI0) protein supported the results of anti-H. pylori activity.

Stable, efficient, and cost-effective system for the biosynthesis of recombinant bacterial cellulose in Escherichia coli DH5α platform.

BACKGROUND: Owing to its remarkable mechanical properties that surpass the plant-based cellulose, bacterial cellulose production has been targeted for commercialization during the last few years. However, the large-scale production of cellulose is generally limited by the slow growth of producing strains and low productivity which ultimately makes the commercial production of cellulose using the conventional strains non cost-effective. In this study, we developed a novel plasmid-based expression system for the biosynthesis of cellulose in E. coli DH5α and assessed the cellulose productivity relative to the typically used E. coli BL21 (DE) expression strain. RESULTS: No production was detected in BL21 (DE3) cultures upon expression induction; however, cellulose was detected in E. coli DH5α as early as 1 h post-induction. The total yield in induced DH5α cultures was estimated as 200 ± 5.42 mg/L (dry weight) after 18 h induction, which surpassed the yield reported in previous studies and even the wild-type Gluconacetobacter xylinum BRC5 under the same conditions. As confirmed with electron microscope micrograph, E. coli DH5α produced dense cellulose fibers with ~ 10 µm diameter and 1000-3000 µm length, which were remarkably larger and more crystalline than that typically produced by G. hansenii. CONCLUSIONS: This is the first report on the successful cellulose production in E. coli DH5α which is typically used for plasmid multiplication rather than protein expression, without the need to co-express cmcax and ccpAx regulator genes present in the wild-type genome upstream the bcs-operon, and reportedly essential for the biosynthesis.

Anticancer, Anticoagulant, Antioxidant and Antimicrobial Activities of Thevetia peruviana Latex with Molecular Docking of Antimicrobial and Anticancer Activities.

Natural origin molecules represent reliable and excellent sources to overcome some medicinal problems. The study of anticancer, anticoagulant, and antimicrobial activities of Thevetia peruviana latex were the aim of the current research. An investigation using high-performance liquid chromatography (HPLC) revealed that the major content of the flavonoids are rutin (11.45 µg/mL), quersestin (7.15 µg/mL), naringin (5.25 µg/mL), and hisperdin (6.07 µg/mL), while phenolic had chlorogenic (12.39 µg/mL), syringenic (7.45 µg/mL), and ferulic (5.07 µg/mL) acids in latex of T. peruviana. Via 1,1-diphenyl-2- picrylhydrazyl (DPPH) radical scavenging, the experiment demonstrated that latex had a potent antioxidant activity with the IC50 43.9 µg/mL for scavenging DPPH. Hemolysis inhibition was 58.5% at 1000 µg/mL of latex compared with 91.0% at 200 µg/mL of indomethacin as positive control. Negligible anticoagulant properties of latex were reported where the recorded time was 11.9 s of prothrombin time (PT) and 29.2 s of the activated partial thromboplastin time (APTT) at 25 µg/mL, compared with the same concentration of heparin (PT 94.6 s and APPT 117.7 s). The anticancer potential of latex was recorded against PC-3 (97.11% toxicity) and MCF-7 (96.23% toxicity) at 1000 µg/mL with IC50 48.26 µg/mL and 40.31 µg/mL, respectively. Disc diffusion assessment for antimicrobial activity recorded that the most sensitive tested microorganisms to latex were Bacillus subtilis followed by Escherichia coli, with an inhibition zone (IZ) of 31 mm with minimum inhibitory concentration (MIC) (10.2 µg/mL) and 30 mm (MIC, 12.51 µg/mL), respectively. Moreover, Candida albicans was sensitive (IZ, 28 mm) to latex, unlike black fungus (Mucor circinelloides). TEM examination exhibited ultrastructure changes in cell walls and cell membranes of Staphylococcus aureus and Pseudomonas aeruginosa treated with latex. Energy scores of the molecular docking of chlorogenic acid with E. coli DNA (7C7N), and Rutin with human prostate-specific antigen (3QUM) and breast cancer-associated protein (1JNX), result in excellent harmony with the experimental results. The outcome of research recommended that the latex is rich in constituents and considered a promising source that contributes to fighting cancer and pathogenic microorganisms.