Garth Maker

Herbal and dietary supplements (HDS) are popular among consumers seeking a 'natural' approach for improving their health; however, at present , there is a lack of evidence to support the claims of efficacy and safety for most of these products. Herbal weight loss supplements (WLS) are a group of HDS that are frequently implicated in cases of toxicity; however, the causative substances often remain unknown due to the complex chemical nature of such supplements. This study aimed to analyse the in vitro safety (in human liver carcinoma (HepG2) cells and colon carcinoma (Caco-2) cells) of 12 active compounds commonly found in WLS, first with safety screening using the MTT cytotoxicity assay, followed by metabolic profiling with 1 H NMR spectroscopy. Of the phytochemicals evaluated, epigallocatechin-3,0-gallate (EGCG) was the only compound that caused a significant reduction in the viability of both cell lines (25.3% in HepG2 cells and 18.5% in Caco-2 cells), and this decrease was potentiated by CYP450 induction with rifampicin. Subsequent 1 H NMR analysis showed changes in key metabolites such as amines, amino acids, carboxylic acids, and glucose that were indicative of protein degradation and disrupted energy and lipid metabolism. While the remaining 11 active compounds analysed did not demonstrate significant toxicity in isolation, these require further assessment to determine their safety when used in combination with other phytochemicals. Given that the majority of WLS contain multiple herbal ingredients, each with a complex chemical composition, it is important to understand the role of interactions in adverse events. Abbreviations ANOVA, analysis of variance; ARTG, Australian Register of Therapeutic Goods; AUROC, area under the receiver operator curve; BBI, broadband decoupling inverse;

Postharvest lipid degradation is a major factor contributing to quality loss, off-flavors, and reduced shelf life in stored rice grains. We investigated whether inducing secondary dormancy using abscisic acid (ABA) can stabilize lipid metabolism in rice. Non-dormant japonica (Nanjing 46) and indica (Fengliangyou) rice grains were induced into secondary dormancy with ABA (800 and 1500 mg/L, respectively), then subjected to accelerated aging (35 °C, 75 ± 2 % RH, up to 45 days). Lipid metabolism was assessed by enzyme assays, UHPLC–MS/MS lipidomics, and RNA-seq. Dormancy induction significantly suppressed the activities of lipase, phospholipase D, and lipoxygenase, resulting in lower rates of lipid hydrolysis and oxidative deterioration. Lipidomic profiling revealed delayed degradation of triacylglycerols and membrane phospholipids (phosphatidylcholine and phosphatidylethanolamine), while transcriptomic analysis showed coordinated downregulation of lipid degradation genes (OsLOX1, OsPLDalpha3). This multi-level suppression led to decreased accumulation of free fatty acids, lipid peroxides, and volatile aldehydes associated with rancid off-flavors. These findings provide new insights into dormancy-mediated lipid regulation and suggest a novel physiological strategy to enhance the storage stability and sensory quality of rice and other lipid-rich cereals.

Background/Objectives: The prevalence of obesity continues to rise globally, and with this an increase in the use of herbal weight loss supplements (WLS). At present, there is limited evidence to support the efficacy and safety of WLS, and there have been growing reports of adverse events associated with their use. We aimed to determine those WLS that caused toxicity in vitro and to use 1H nuclear magnetic spectroscopy (NMR) to examine the metabolomic changes induced by these WLS in human hepatic and intestinal cells. Materials and Methods: This study used in vitro methods and 1H NMR spectroscopy to analyse the metabolomic changes in vitro of WLS available for purchase in Australia. Ten WLS were selected, nine WLS caused significant toxicity in HepG2 human liver cells, and of these, six met the criteria for 1H NMR analysis, which was based on a 25–50% reduction in cell viability. Results: All 10 WLS caused a significant reduction in viability of Caco-2 human intestinal cells, with seven selected for metabolic profiling. Orthogonal partial least squares discriminant analysis (O-PLS-DA) of 1H NMR spectral data was used to characterise the metabolites that differed between the untreated and treated cells and the fold changes of the metabolites were determined. The results showed alterations to key metabolites such as amino acids, glucose, carboxylic acids, and amines in all treatment groups compared to untreated controls across both cell lines. Conclusions: Collectively, these biochemical changes represent disturbances to intracellular proteins, energy metabolism, and membrane lipids suggestive of oxidative stress. This study highlights the need for further investigations into the actions of these WLS in vivo, and, as these products were regulated by the Therapeutic Goods Administration (TGA) at the time of purchase, this study suggests improved pre-market screening to ensure consumer health is protected.

Cuticular hydrocarbons (CHCs) in blow flies vary with species, age, and environmental factors. Blow fly puparia are often found associated with decomposing remains, but their utility in post-mortem interval (PMI) estimation in forensic cases is limited by challenges in accurately ageing them using conventional methods. This study builds on prior research by successfully distinguishing the CHC profiles of Lucilia sericata full puparia aged 1- and 5- days post-pupation. The results indicate that CHC analysis could serve as a valuable tool, either as a complement to or as a potential alternative for traditional forensic entomology methods.

Furocoumarins (FCs), important natural compounds with biodefense roles and pharmacological activities, are notably abundant in medicinal plant Angelica dahurica. However, its accumulation patterns over development stages in FC-enriched tissue, biosynthetic pathways, and regulatory mechanisms in A. dahurica remain elusive. Here, we quantified the concentration dynamics of 17 coumarins across six developmental stages of root and found a gradual decrease in FC concentration as the roots develop. Using a de-novo assembled chromosome-level genome for A. dahurica, we conducted integrative multi-omics analyses to screen out candidate genes to fill in the sole missing step in the biosynthesis of imperatorin and isoimperatorin. This revealed that CYP71AZ18 catalyzes hydroxylation at the C-5 position of psoralen to generate bergaptol, while CYP71AZ19 and CYP83F95 catalyze hydroxylation at the C-8 position to produce xanthotoxol, notably indicating that a single step is catalyzed by two genes from distinct CYP450 subfamilies in this species. CYP71AZ19 originated from a proximal duplication event of CYP71AZ18, specific to A. dahurica, and subsequently underwent neofunctionalization. Accessible chromatin regions (ACRs), especially proximal ACRs, correlated with high gene expression levels, and the three validated genes exhibited strong signals of ACRs, showing the importance of chromosomal accessibility in regulating metabolite biosynthesis.

Cuticular hydrocarbons (CHCs) are long-chain lipids found on the exoskeletons of insects, serving primarily as a protective barrier against water loss and environmental factors. In the last few decades, the qualitative and quantitative analysis of CHCs, particularly in blow flies, has emerged as a valuable tool in forensic entomology, offering promising potential for species identification and age estimation of forensically important insects. This review examines the current application of CHC analysis in forensic investigations and highlights the significant advancements in the field over the past few years. Studies have demonstrated that CHC profiles vary with insect development, and while intra-species variability exists due to factors such as age, sex, geographical location, and environmental conditions, these variations can be harnessed to refine post-mortem interval (PMI) estimations and improve the accuracy of forensic entomological evidence. Notably, CHC analysis can also aid in distinguishing between multiple generations of insects on a body, providing insights into post-mortem body movement and aiding in the interpretation of PMI in complex cases. Furthermore, recent studies have investigated the variability and degradation of CHCs over time, revealing how environmental factors-such as temperature, humidity, UV light exposure, and toxicological substances-affect CHC composition, providing valuable insights for forensic investigations. Despite the promise of CHC profiling, several challenges remain, and this review also aims to highlight future research directions to enhance the reliability of this technique in forensic casework.

Whilst wound repair in severe burns has received substantial research attention, non-severe burns (<20% total body surface area) remain relatively understudied, despite causing considerable physiological impact and constituting most of the hospital admissions for burns. Early prediction of healing outcomes would decrease financial and patient burden, and aid in preventing long-term complications from poor wound healing. Lipids have been implicated in inflammation and tissue repair and may play essential roles in burn wound healing. In this study, plasma samples were collected from 20 non-severe burn patients over 6 weeks from admission, including surgery, and analysed by liquid chromatography-tandem mass spectrometry and nuclear magnetic resonance spectroscopy to detect 850 lipids and 112 lipoproteins. Orthogonal projections to latent structures-discriminant analysis was performed to identify changes associated with re-epithelialisation and delayed re-epithelisation.
We demonstrated that the lipid and lipoprotein profiles at admission could predict re-epithelisation outcomes at 2 weeks post-surgery, and that these discriminatory profiles were maintained up to 6 weeks post-burn. Inflammatory markers GlycB and C-reactive protein indicated divergent systemic responses to the burn injury at admission. Triacylglycerols, diacylglycerols and low-density lipoprotein subfractions were associated with delayed wound closure (p-value <0.02, Cliff’s delta >0.7), whilst high-density lipoprotein subfractions, phosphatidylinositols, phosphatidylcholines, and phosphatidylserines were associated with re-epithelisation at 2 weeks post-surgery (p-value <0.01, Cliff’s delta <-0.7). Further model validation will potentially lead to personalised intervention strategies to reduce the risk of chronic complications post-burn injury.
Abbreviations

AUROCArea under receiver operating characteristicsCABINRandomised placebo-controlled trial of Celecoxib for Acute Burn Inflammation and FeverCRPC-reactive proteinCVCross-validatedDIREDiffusion and relaxation editingDGDiacylglycerolDREDelayed wound re-epithelisationFAFatty acylHDLHigh-density lipoproteinIDLIntermediate-density lipoproteinIPAIsopropyl alcoholIQRInter-quartile rangeLC-QQQ-MSLiquid chromatography-tandem mass spectrometryLDCHLow-density lipoprotein cholesterolLDHDLow-density lipoprotein / high-density lipoprotein ratioLDLLow-density lipoproteinLDPLLow-density lipoprotein phospholipidsLPILysophosphatidylinositolNMRNuclear magnetic resonanceOPLS-DAOrthogonal projections to latent structures-discriminant analysisPCPhosphotidylcholinePCAPrincipal component analysisPGPhosphatidylglycerolPIPhosphatidylinositolPLSPartial least squaresPOSASPatient and observer scar assessment scalePSPhosphotidylserineQCQuality controlQTRAPQuadrupole ion trapREWound re-epithelisationSDStandard deviationTBSATotal body surface areaTGTriacylglyerolVLDLVery low-density lipoprotein

The immortalised human hepatocellular HepG2 cell line is commonly used for toxicology studies as an alternative to animal testing due to its characteristic liver-distinctive functions. However, little is known about the baseline metabolic changes within these cells upon toxin exposure. We have applied high-resolution 1H Nuclear Magnetic Resonance (NMR) spectroscopy to characterise the biochemical composition of HepG2 cells at baseline and post-exposure to hydrogen peroxide (H2O2). Metabolic profiles of live cells, cell extracts, and their spent media supernatants were obtained using 1H high-resolution magic angle spinning (HR-MAS) NMR and 1H NMR spectroscopic techniques. Orthogonal partial least squares discriminant analysis (O-PLS-DA) was used to characterise the metabolites that differed between the baseline and H2O2 treated groups. The results showed that H2O2 caused alterations to 10 metabolites, including acetate, glutamate, lipids, phosphocholine, and creatine in the live cells; 25 metabolites, including acetate, alanine, adenosine diphosphate (ADP), aspartate, citrate, creatine, glucose, glutamine, glutathione, and lactate in the cell extracts, and 22 metabolites, including acetate, alanine, formate, glucose, pyruvate, phenylalanine, threonine, tryptophan, tyrosine, and valine in the cell supernatants. At least 10 biochemical pathways associated with these metabolites were disrupted upon toxin exposure, including those involved in energy, lipid, and amino acid metabolism. Our findings illustrate the ability of NMR-based metabolic profiling of immortalised human cells to detect metabolic effects on central metabolism due to toxin exposure. The established data sets will enable more subtle biochemical changes in the HepG2 model cell system to be identified in future toxicity testing.The immortalised human hepatocellular HepG2 cell line is commonly used for toxicology studies as an alternative to animal testing due to its characteristic liver-distinctive functions. However, little is known about the baseline metabolic changes within these cells upon toxin exposure. We have applied high-resolution 1H Nuclear Magnetic Resonance (NMR) spectroscopy to characterise the biochemical composition of HepG2 cells at baseline and post-exposure to hydrogen peroxide (H2O2). Metabolic profiles of live cells, cell extracts, and their spent media supernatants were obtained using 1H high-resolution magic angle spinning (HR-MAS) NMR and 1H NMR spectroscopic techniques. Orthogonal partial least squares discriminant analysis (O-PLS-DA) was used to characterise the metabolites that differed between the baseline and H2O2 treated groups. The results showed that H2O2 caused alterations to 10 metabolites, including acetate, glutamate, lipids, phosphocholine, and creatine in the live cells; 25 metabolites, including acetate, alanine, adenosine diphosphate (ADP), aspartate, citrate, creatine, glucose, glutamine, glutathione, and lactate in the cell extracts, and 22 metabolites, including acetate, alanine, formate, glucose, pyruvate, phenylalanine, threonine, tryptophan, tyrosine, and valine in the cell supernatants. At least 10 biochemical pathways associated with these metabolites were disrupted upon toxin exposure, including those involved in energy, lipid, and amino acid metabolism. Our findings illustrate the ability of NMR-based metabolic profiling of immortalised human cells to detect metabolic effects on central metabolism due to toxin exposure. The established data sets will enable more subtle biochemical changes in the HepG2 model cell system to be identified in future toxicity testing.

Drink spiking in social settings is one of the most pervasive forms of drug-facilitated sexual assault (DFSA). There are no current data in Australia on the rates of drink spiking or their associated assaults. There is also little known about the prevalence of different substances involved and how the current substance use trends compare to sexual assault trends. To explore this, a recalculation of sexual assault trends to estimate substance related sexual assault was performed. Data about recent trends of mental health prescriptions and sexual assault were obtained from the Australian Institute of Health and Welfare (AIHW). The analysis of these datasets highlighted that females are the highest consumers of antidepressants and benzodiazepines. Results also indicated a statistically significant positive correlation between females and a higher incidence of sexual assault (r = 0.996, p < .001). This paper demonstrates that females are at most risk of drug-drug interactions (e.g., diazepam and ketamine) with their medications due to the higher rate of prescriptions amongst this population, and therefore more vulnerable to both opportunistic and proactive DFSA. While these findings are preliminary and not causal, they highlight trends in need of further study.

The immortalised human hepatocellular HepG2 cell line is commonly used for toxicology studies as an alternative to animal testing due to its characteristic liver-distinctive functions. However, little is known about the baseline metabolic changes within these cells upon toxin exposure. We have applied 1H Nuclear Magnetic Resonance (NMR) spectroscopy to characterise the biochemical composition of HepG2 cells at baseline and post-exposure to hydrogen peroxide (H2O2). Metabolic profiles of live cells, cell extracts, and their spent media supernatants were obtained using 1H high-resolution magic angle spinning (HR-MAS) NMR and 1H NMR spectroscopic techniques. Orthogonal partial least squares discriminant analysis (O-PLS-DA) was used to characterise the metabolites that differed between the baseline and H2O2 treated groups. The results showed that H2O2 caused alterations to 10 metabolites, including acetate, glutamate, lipids, phosphocholine, and creatine in the live cells; 25 metabolites, including acetate, alanine, adenosine diphosphate (ADP), aspartate, citrate, creatine, glucose, glutamine, glutathione, and lactate in the cell extracts, and 22 metabolites, including acetate, alanine, formate, glucose, pyruvate, phenylalanine, threonine, tryptophan, tyrosine, and valine in the cell supernatants. At least 10 biochemical pathways associated with these metabolites were disrupted upon toxin exposure, including those involved in energy, lipid, and amino acid metabolism. Our findings illustrate the ability of NMR-based metabolic profiling of immortalised human cells to detect metabolic effects on central metabolism due to toxin exposure. The established data sets will enable more subtle biochemical changes in the HepG2 model cell system to be identified in future toxicity testing.