Effect of Ducrosia anethifolia methanol extract against methicillin resistant Staphylococcus aureus and Pseudomonas aeruginosa biofilms on excision wound in diabetic mice.

Background: Ducrosia anethifolia is an aromatic desert plant used in Saudi folk medicine to treat skin infections. It is widely found in Middle Eastern countries. Methods: A methanolic extract of the plant was prepared, and its phytoconstituents were determined using LC-MS. In-vitro and in-vivo antibacterial and antibiofilm activities of the methanolic extract were evaluated against multidrug-resistant bacteria. The cytotoxic effect was assessed using HaCaT cell lines in-vitro. Diabetic mice were used to study the in-vivo antibiofilm and wound healing activity using the excision wound method. Results: More than 50 phytoconstituents were found in the extract after LC-MS analysis. The extract exhibited antibacterial activity against both the tested pathogens. The extract was free of irritant effects on mice skin, and no cytotoxicity was observed on HaCaT cells with an IC50 value of 1381 µg/ml. The ointment formulation of the extract increased the healing of diabetic wounds. The microbial load of both pathogens in the wounded tissue was also reduced after the treatment. The extract was more effective against methicillin-resistant Staphylococcus aureus (MRSA) than MDR-P. aeruginosa in both in vitro and in vivo experiments. Further, skin regeneration was also observed in histological studies. Conclusions: The results showed that D. anethifolia methanol extract supports wound healing in infected wounds in diabetic mice through antibacterial, antibiofilm, and wound healing activities.

In Vitro Antiproliferative Activity and Phytochemicals Screening of Extracts of the Freshwater Microalgae, Chlorochromonas danica.

The present study was focused on the screening of phytochemicals, their quantitative estimation and analysis by LC-MS profile, and antiproliferative efficacy of the aqueous-ethanolic extracts of the microalgae, Chlorochromonas danica isolated from the freshwater body Tavanampalli. The aqueous-ethanol extract of Chlorochromonas danica showed the presence of flavonoids, phenols, and proteins. The total flavonoid content, total phenol content, and total protein content were determined to be 158.65 mg of quercetin equivalent, 15.75 mg of gallic acid equivalent, and 134.65 mg/g dry weight of the extract, respectively. The LC-MS analysis confirmed the presence of several major bioactive molecules including L-Histidine, D-glutamine, L-aspartic acid, adenine, adenosine, cotinine, guanine hypoxanthine, L-glutamic acid, nicotinamide, 4-Hydroxycoumarin, and Stearamide. The aqueous-ethanol extract of Chlorochromonas danica exhibited an IC50 values of 63.34 µg, 279.29 µg, 125.42 µg, 90.56 µg, and 95.58 µg against A375, A549, HeLa, HepG2, and HT29 cell lines respectively, compared to the positive control cisplatin with IC50 values of 3.56 µg, 4.65 µg, 3.88 µg, 4.87 µg, and 7.23 µg respectively. These data suggest that Chlorochromonas danica remains a promising drug candidate for the treatment of cancers, particularly melanoma (A375 cell line) that can be considered for purification of antiproliferative compound and further clinical trials for the discovery of novel antiproliferative drugs from cost-effective sources.

Insights into ultrasound-promoted degradation of naphthenic acid compounds in oil sands process affected water. Part I: Accelerated H-abstraction and decarboxylation of aromatic and alicyclic compounds.

Propelled by enormous increase in demand for fuel sources, Canadian oil sands are becoming increasingly important as a fuel source due to their abundance and upgrading capability. However, extraction of bitumen, a high acid crude (HAC) oil, requires 2-3 units of water per unit of oil resulting in naphthenic acid (NA)-rich oil sands process affected water (OSPW) collected in effluent ponds. This study illustrates the role of sonochemistry in the accelerated degradation through H-abstraction and subsequent decarboxylation of aromatic and alicyclic naphthenic acid model compounds. Benzoic acid and 3-methylcyclohexane carboxylic acid were selected as model NA compounds to investigate the mechanism of hydroxyl radical (OH•) initiated carboxylic acid degradation in 378 KHz sonochemical reactor. Established FTIR methods with low resolution LCMS spectroscopy confirmation were applied to determine the extent of carboxylic acid degradation and identify the formation of products. FTIR monitoring showed a non-linear degradation of carboxylic acids with formation of many intermediates highlighting the shift from cyclic carboxylic acids to cyclic alcohols during BA degradation. Subsequent decrease in carboxylic acid groups signifies scission of cyclic structures before complete mineralization. This is confirmed with the LCMS identification of products such as: 3-hydroxybenzoic acid and phenol. This study postulated new breakdown pathways for degradation of benzoic acid with complete mineralization at a sonochemical reaction time (SRT) of 4 h. A radical quenching process was also inferred through the formation of conglomerates during sonochemical degradation of BA. Extension of the study to 3-methylcyclohexane carboxylic acid (3mCHA) shows similar non-linearity with an increase in carboxylic acid groups indicating H-abstraction followed by ring-opened compounds. However, due to the complex nature of 3mCHA's ring-opened compounds, complete mineralization is not achieved. The putative role of sonochemistry is a promising and sustainable degradation method for mitigating NAs in OSPW, but sonication periods need to be considered carefully to ensure adequate mineralization of their constituents and combinatorial methods with other advanced oxidation methods may be needed to enhance industrial application. In Part II, an in silico screening approach using first principles is reported to identify the breakdown of the organic compounds and determine molecular rates of reaction to confirm the mechanistic origins of the compounds formed.

Phytochemical profiling of Azima tetracantha Lam. leaf methanol extract and elucidation of its potential as a chain-breaking antioxidant, anti-inflammatory and anti-proliferative agent.

Azima tetracantha, a traditional medicinal plant included in the order Brassicales and family Salvadoraceae, is widely used as a dietary supplement in folklore medicines. The plant is also used for the treatment of rheumatism, diarrhea and other inflammatory disorders. The present investigation focused on the phytochemical composition, radical scavenging, reducing potential and anti-proliferative activities of the A. tetracantha leaves. Quantitative estimation of the polyphenols and flavonoids revealed significantly elevated levels in the methanol extract. Corroborating with this, methanol extract exhibited higher in vitro anti-radical scavenging effect against 2,2-diphenyl-1- picrylhydrazyl (34.14 ± 2.19 µg/mL), and hydrogen peroxide (44.96 ± 1.77 µg/mL), as well as ferric reducing properties (58.24 ± 6.98 µg/mL). The methanolic extract also showed strong lipoxygenase (71.42 ± 6.36 µg/mL) and nitric oxide inhibitory activities (94.23 ± 8.11 µg/mL). Cytotoxic activity against MCF7 cells was found to be higher (IC50= 37.62 ± 2.94 µg/mL), than that of MDAMB231 cells (IC50= 69.11 ± 5.02 µg/mL). The qPCR-based analysis indicated dose-dependent increase in the expression of the pro-apoptotic genes such as executioner caspases and apoptotic protease activating factor-1. Overall, the results indicated the possible use of methanol extract of A. tetracantha leaves as a chain-breaking antioxidant molecule and are capable of inhibiting inflammatory enzymes and the proliferative potential of breast cancer cells.

Serum markers in diagnostics of steatohepatitis.

Along with the increasing incidence and prevalence of obesity and the metabolic syndrome, the number of patients with its hepatic manifestation - NAFL, characterized by triglyceride storage in liver, is rising. NAFL (non-alcoholic fatty liver) is now, with the prevalence of 40 %, the most common liver disease in Western countries. Despite that NAFL has usually no symptoms and in most patients, it is diagnosed as an incidental finding by abdominal ultra-sound, every third of these patients develops NASH (non-alcoholic steatohepatitis), resulting in an individual progression of the sequence of fibrosis - cirrhosis - hepatocellular carcinoma. Due to the fact, that NASH is, along with the cardiovascular causes, involved in liver-related mortality of patients with the metabolic syndrome, from clinical view, it is fundamental to distinguish between benign NAFL and potentially progressive NASH. This appears even more serious realizing that patients with NASH are being often underdiagnosed because of limited indications of liver biopsy, a common diagnostically gold standard. This work emphasizes the relationship between metabolic syndrome and liver disease and presents the main diagnostic possibilities of NAFL/NASH, the most dealing with serum markers. It is based on a research, using the PubMed database and putting the key words as search terms. Considering the huge number of patients diagnosed with fatty liver, a non-invasive, widely approachable method should be established, to make the diagnostic and staging of progression of NASH broadly possible. A new method using LCMS (Liquid Chromatography-Mass Spectrometry) analysis of serum lipids now fulfils these criteria, having high enough specificity and sensitivity, and have also been validated by comparing with a large cohort of patients diagnosed with liver biopsy.