The challenges of the COVID-19 pandemic have highlighted an increasing clinical demand for safe and effective treatment options against an overzealous immune defence response, also known as the ..."cytokine storm". Andrographolide is a naturally derived bioactive compound with promising anti-inflammatory activity in many clinical studies. However, its cytokine-inhibiting activity, in direct comparison to commonly used nonsteroidal anti-inflammatory drugs (NSAIDs), has not been extensively investigated in existing literature. The anti-inflammatory activities of andrographolide and common NSAIDs, such as diclofenac, aspirin, paracetamol and ibuprofen were measured on lipopolysaccharide (LPS) and interferon-gamma induced RAW264.7 cells. The levels of PGE2, nitric oxide (NO), TNF-alpha & LPS-induced release of pro-inflammatory cytokines on differentiated human macrophage THP-1 cells were measured against increasing concentrations of andrographolide and aforementioned NSAIDs. The associated mechanistic pathway was examined on NFkappaB using flow cytometry on the human endothelial-leukocyte adhesion molecule (ELAM9) (E-selectin) transfected RAW264.7 cells with green fluorescent protein (GFP). Andrographolide exhibited broad and potent anti-inflammatory and cytokine-inhibiting activity in both cell lines by inhibiting the release of IL-6, TNF-alpha and IFN-gamma, which are known to play a key role in the etiology of cytokine storm and the pathogenesis of inflammation. In comparison, the tested NSAIDs demonstrated weak or no activity against proinflammatory mediators except for PGE2, where the activity of andrographolide (IC.sub.50 = 8.8 muM, 95% CI = 7.4 to 10.4 muM) was comparable to that of paracetamol (IC.sub.50 = 7.73 muM, 95% CI = 6.14 to 9.73 muM). The anti-inflammatory action of andrographolide was associated with its potent downregulation of NFkappaB. The wide-spectrum anti-inflammatory activity of andrographolide demonstrates its therapeutic potential against cytokine storms as an alternative to NSAIDs.
This study evaluates the in vitro antioxidant and immunomodulation activities of essential oils isolated from an anti-upper respiratory tract infection (URTI) formulation with a view to their ...therapeutic potential. The chemical components of the essential oil were analysed by gas chromatography-mass spectrometry (GC-MS). The antioxidative activity of the oils was investigated with regard to their ability to scavenge DPPH●, ABTS●+, and hydroxyl free radical (•OH). Their immunostimulatory activities were determined using murine macrophage cells. The main components of the oil with pharmacological and biological activities include 1,8-eucalyptol (42.9%), patchouli alcohol (19.9%), trans-erinolide (9.2%), and guaiacol (5%). The oils displayed high DPPH, ABTS, and hydroxyl radical scavenging activities and anti-inflammatory activities by reducing tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) production. The results indicate that essential oils have the potential to be used in products for anti-URTI treatment.
This study aims to determine the relationship between the antioxidant and anti-inflammatory activities of the thirteen herbs and two fungi extracts, and their total phenolic and flavonoid contents.
...Antioxidant activities were evaluated by four assays: an antioxidant activity assay using Saccharomyces cerevisiae, a DPPH ((2, 2-diphenyl-1-picrylhydrazyl) assay to assess free radical scavenging, an assay assessing ferrous ions or iron (II) chelating ability, and a ferric reducing antioxidant power (FRAP) assay. Total phenolic and flavonoid contents were determined using the Folin-Ciocalteu and aluminium chloride methods, respectively. Anti-inflammatory activities were determined by measuring the inhibition of nitric oxide and TNF-α production in lipopolysaccharide- and interferon-γ-activated J774A.1 macrophages. Their cytotoxicities against macrophages were determined by MTT assay.
A positive linear correlation between antioxidant activities and the total phenolic and flavonoid content of the plant extracts was found. The plant extracts with high phenolic and flavonoid content also exhibited significant anti-inflammatory activity with good cell viability.
The selected herbs could be a rich source of antioxidants and free radical scavenging compounds. The levels of phenolic and flavonoid compounds were correlated with the antioxidant and anti-inflammatory activities of the extracts from the herbs.
Arimisia annua L. is an important anticancer herb used in traditional Chinese medicine. The molecular basis underpinning the anticancer activity is complex and not fully understood, but the herbal ...polysaccharides, broadly recognised as having immunomodulatory, antioxidant and anticancer activities, are potential key active agents. To examine the functions of polysaccharides from A. annua, their immunomodulatory and antioxidant potentials were evaluated, as well as their structural characterization. The water-soluble polysaccharides (AAPs) were fractionated using size-exclusion chromatography to obtain three dominant fractions, AAP-1, AAP-2 and AAP-3, having molecular masses centered around 1684, 455 and 5.8kDa, respectively. The antioxidant potentials of the isolated polysaccharides were evaluated by measuring radical scavenging activities against DPPH● (2,2-diphenyl-1-picrylhydrazyl radical), ABTS●+ (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid radical ion), and the OH● (hydroxyl radical). AAP-1 displayed high antioxidant activities against these radicals, which were 68%, 73% and 78%, respectively. AAP-2 displayed lower scavenging activities than the other two fractions. Immunostimulatory activities of AAPs were measured using mouse macrophages. The three polysaccharide fractions displayed significant antioxidant activities and stimulated the production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). AAP-1 showed significant immunostimulatory activity (16-fold increase in the production of IL-6 compared to the control and 13-fold increase in the production of TNF-α) with low toxicity (>60% cell viability at 125 μg/mL concentration). Preliminary structural characterization of the AAPs was carried out using gas chromatography (GC) and FTIR techniques. The results indicate that AAP-1 and AAP-2 are pyranose-containing polysaccharides with β-linkages, and AAP-3 is a β-fructofuranoside. The results suggest that these polysaccharides are potential candidates for immunotherapy and cancer treatment.
The Naoxinqing (NXQ) tablet is a standardised proprietary herbal product containing an extract of persimmon leaves (
) for the management of cardio- and cerebrovascular diseases. Although previous ...reports suggested that the efficacy of NXQ is at least partly mediated by its anti-oxidative property, the anti-oxidative effect of the major components of NXQ has not been studied systematically. For quality control purposes, only analytical methods limited to 3 marker analytes have been reported, the extent to which the other components affect efficacy has not been explored. In this study, we developed an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC MS/MS) method for the identification of seven analytes (kaempferol-3-
-glucoside (astragalin), quercetin-3-
-galactoside (hypericin), quercetin-3-
-glucoside (isoquercitin), kaempferol, 3,4-dihydroxybenzoic acid (protocatechuic acid), and furan-2-carboxylic acid (pyromucic acid) and quercetin) in the NXQ. This is the first method reported and validated for the quantification of the seven major secondary metabolites in NXQ. The results for the quantified analytes were then compared in 15 different batches of NXQ. The variation observed in the seven components highlights the need to quantify key bioactive components to ensure product consistency. Radical scavenging activity and abundance was used to rank the analytes. The anti-oxidative effects of NXQ were examined using cultured human vascular endothelial cells (EA.hy926). Corrected 2,2-di(4-tert-octylphenyl)-1-picrylhydrazyl (DPPH) activity results revealed that quercetin and kaempferol have the strongest anti-oxidant capacity in the extract. Both quercetin and kaempferol significantly inhibited the hydrogen peroxide (H₂O₂)-induced EA.hy926 cell injury and intracellular reactive oxygen species (ROS) generation. In conclusion, we established and validated an UPLC-MS/MC method for the analysis of major bioactive components in the NXQ and demonstrated that its anti-oxidative property may play a critical role in cerebrovascular protection.
Background-The quality control (QC) for commercial herbal formulations is sparse due to a lack of well-developed HPLC-ESI-MS/MS methods. Objective-This study reports the quantification of nine ...selected analytes for a commercial eight-herb formulation known as Qi Ju Di Huang Wan (QJDHW) used to relieve hypertension. Methods-An HPLC-ESI/MS method for the quantitation of analytes selected using the Herbal Chemical Marker Ranking System (Herb MaRS) was developed. The Herb MaRS ranking system which takes into account bioavailability, bioactivity, and physiological action related to its intended use and the commercial availability of the standard. After a method optimization, seven analytes were found to be ideal for quantitation. Results-The target analytes were identified using an electrospray ionization-tandem MS molecular breakdown comparison between the herbal peak and the commercial standard. The quantitative aspect of analyte variability of eleven samples was studied using fold variation. The fold variation of selected analytes among eleven samples ranged from 1.5 to 28.9. The qualitative aspect of variability was studied using principal component analysis (PCA) and hierarchical cluster analysis (HCA). Conclusions-There is a great degree of chemical variability in herbal formulations which are due to raw material harvesting times, storage techniques, and plant subspecies variability. Highlights-Commercial QJDHW formulations need to be standardised using HPLC-ESI-MS/MS to ensure better product quality control (QC) and product efficacy for the consumer.
To compare health and science students' demographic characteristics and learning approaches across different disciplines, and to examine the relationship between learning approaches and academic ...performance.
While there is increasing recognition of a need to foster learning approaches that improve the quality of student learning, little is known about students' learning approaches across different disciplines, and their relationships with academic performance.
Prospective, correlational design.
Using a survey design, a total of 919 first year health and science students studying in a university located in the western region of Sydney from the following disciplines were recruited to participate in the study — i) Nursing: n=476, ii) Engineering: n=75, iii) Medicine: n=77, iv) Health Sciences: n=204, and v) Medicinal Chemistry: n=87.
Although there was no statistically significant difference in the use of surface learning among the five discipline groups, there were wide variations in the use of deep learning approach. Furthermore, older students and those with English as an additional language were more likely to use deep learning approach. Controlling for hours spent in paid work during term-time and English language usage, both surface learning approach (β=−0.13, p=0.001) and deep learning approach (β=0.11, p=0.009) emerged as independent and significant predictors of academic performance.
Findings from this study provide further empirical evidence that underscore the importance for faculty to use teaching methods that foster deep instead of surface learning approaches, to improve the quality of student learning and academic performance.
•Relative response factors are valid for the HPLC-PDA analysis of ten cannabinoids.•Quantification was achieved without costly standards, only requiring CBD and CBDA.•Surrogate matrices were valid ...for determining the method accuracy by spike-recovery.
The quality control of medicinal cannabis should include quantification of as many cannabinoids as practicable in a routine analytical lab, to accurately reflect the quality of the product. However, the cost and availability of some cannabinoid standards is an impediment to their routine use. This work seeks to overcome this obstacle by analysing samples using relative retention times (RRT) and relative response factors (RRF), relative to CBD and CBDA reference standards which are readily available. A high-performance liquid chromatography-photodiode array method was developed to quantify ten cannabinoids (Δ9-THC, Δ8-THC, THCA-A, CBN, CBD, CDBA, CBC, CBDV, CBG, and CBGA) in dried cannabis inflorescence and cannabis oil. This method was validated according to ICH guidelines. The proposed method has detection limits ranging from 20 to 78 µg/g, which provided sufficient sensitivity for the panel of cannabinoids. Non-cannabinoid surrogate matrices were used for spike recovery studies to determine method accuracy – analyte recoveries for the inflorescence and oil ranged from 90.1 to 109.3% (inflorescence mean, 100.9%; oil mean, 99.6%). The RRT and RRF values determined independently by three analysts were comparable, indicating the method is robust. The validity of analysis using RRT and RRF was further confirmed by testing six inflorescence samples, as it was found that concentrations above the order of magnitude of the LoQ agreed satisfactorily (range, 95.0 to 111.9%; mean, 100.0%) with the concentrations obtained through the conventional approach of multipoint calibration using pure standards. The proposed method is therefore suitable for the rapid and simple determination of a panel of ten cannabinoids without having to repeatedly purchase every expensive pure standard. Accordingly, analysts in the medicinal cannabis field may explore the use of RRF and RRT for their methods and instruments.
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Amauroderma rugosum (Blume and T. Nees) Torrend is a traditionally well-known mushroom that is used for the treatment of cancer. In order to evaluate the pharmacological activities of A. rugosum ...polysaccharides, the mushroom powder was subjected to hot water extraction and pure polysaccharides (ARPs) were isolated by gel-filtration method. Three important APRs called ARP-1, ARP-2 and ARP-5 were identified with average molecular weights of 1494, 450, and 7 kDa respectively. Their antioxidant abilities were estimated by examining free radical scavenging potential against 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid radical (ABTS●+), 2,2-diphenyl-1-picrylhydrazyl radical (DPPH●), and hydroxyl radical. Immunomodulatory potentials of these ARPs were determined using murine macrophage cells. These polysaccharides exhibited high antioxidant abilities and stimulated mouse macrophages leading to the generation of tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Excellent activities were displayed by ARP-1 and APR-2. Gas chromatography and spectroscopic (FT-IR and NMR) methods were employed in order to carry out their structural characterisation. The two high molecular weight ARPs (ARP-1 and ARP-2) displayed β-(1 → 3)-D-glucan backbone structure with branching of β-(1 → 6)-d-glucopyranosyl. These observations suggest high potential of ARPs for immunotherapeutic applications.
•Five polysaccharides have been isolated from Amauroderma rugosum.•Amauroderma rugosum Polysaccharides exhibit significant immunostimulatory activities and low toxicity.•Two of the identified Amauroderma rugosum Polysaccharides have high potential to develop immunotherapeutic agents.•Two active polysaccharides (ARP-1 and ARP-2) exhibit β-(1. → 3)-D-glucan backbone structure with branching of β-(1 → 6)-d-glucopyranosyl
There is a need for increased QC of complex herbal medicine formulations to ensure product consistency, efficacy, and safety. This study reports an HPLC with photodiode array and electrospray ...ionization-tandem MS method for quantifying selected analytes in a seven-herb formulation. Fourteen analytes were selected for quantification based on the criteria available from the Herbal Chemical Marker Ranking System, which takes into account the bioavailability, reported bioactivity, and physiological action related to its intended use, as well as commercial availability of the standard. After optimizing the columns and chromatographic conditions, 13 of the 14 analytes were able to be determined in one run, with the remaining analyte analyzed on its own. The method was successfully applied to two different extracts of the formulation, demonstrating an application for the QC of a complex herbal mixture with respect to their chemical characteristics.
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