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Sourcing in chemical forensic science refers to the attribution of a sample to a specific source using a characteristic signature. It relies on the identification of chemical attribution signatures (CAS), including chemical markers such as residual synthetic precursors, impurities, reaction by-products and degradation products, or even metabolites. Undertaking CAS for chemical threat agents (CTA) can be used to provide an evidentiary link between the use of a given chemical and its precursor(s) to support forensic investigations. Organophosphorus compounds, a class of nerve agents, can be produced by different, more or less complex synthesis routes that can lead to specific CAS. Chlorpyrifos (CPF), an organophosphorus pesticide, was selected as model compound. To assess the specificity of impurity markers originated from a chemical synthesis, untargeted fingerprints of crude CPF from different synthesis pathways were analyzed as a first use-case using metabolomics-based trace discovery strategies. Seven different CPF synthesis routes were considered, and their crude mixtures were analyzed with a minimal sample preparation. Analyses were performed on a trapped ion mobility spectrometry (TIMS) coupled to liquid chromatography (LC) and high-resolution mass spectrometry (HRMS). Chemometrics analyses were conducted with multivariate methods to extract discriminating features (i.e., relevant impurities), annotate, and identify them. Then, unknown samples were analyzed in blind conditions without any information of the synthesis pathway employed. The aim is to validate the methodology seeking some discriminating impurities identified in the first section to attribute and classify them according to the synthesis route., (© 2025 John Wiley & Sons Ltd.)

Adduction on protein nucleophile sites by mustard agents can be monitored to assess detection of retrospective exposure to these agents. Cysteine 34 (Cys34) on human serum albumin was selected as the target of choice. This work targets di- and tripeptides adducted on Cys34 by sulfur mustard, sesquimustard, and nitrogen mustards separated in hydrophilic liquid chromatography (HILIC) and Reversed-Phase (RP) mode. The effect of several mobile phase additives on the mass spectrometry (MS) and MS/MS signal and on LC retention profile was studied. A mix of ammonium acetate and acetic acid offered satisfactory results in terms of MS sensitivity. Screening of HILIC columns was performed, and ZIC-HILIC stationary phase was selected for HILIC mode, and C 18 stationary phase was used for RP analysis. Negative ionization mode leads to a higher S/N ratio compared to positive ionization mode. Adducted tripeptides were selected for the monitoring of mustard agents' exposure, allowing better sensitivity than their dipeptide homologues. The two developed chromatographic methods have similar sensitivities with LOQs ranging from 1.9 to 20.5 ng/mL for the reversed-phase liquid chromatography (RPLC)-ESI-(-)-MS/MS method and from 1.7 to 43.3 ng/mL for the HILIC-ESI-(-)-MS/MS method. The monitoring method should be selected based on the targeted mustard agent, and the remaining method can be a confirmation tool., (© 2024 Wiley‐VCH GmbH.)

Hyperemesis gravidarum (HG) is a severe condition marked by intense nausea and vomiting during pregnancy, which is different from typical morning sickness. It is marked by weight loss exceeding 5% of pre-pregnancy weight, ketonuria, dehydration, electrolyte imbalances, and in some cases, arrhythmias - primarily linked to electrolyte disturbances. Treatment typically involves conservative measures such as small, bland meals, medications like metoclopramide and ondansetron, and correction of electrolyte abnormalities. This case study presents a 29-year-old female, G5P2022, who arrived at the ED with severe nausea, vomiting, intermittent chest pain, and palpitations lasting five days. She was confirmed to be seven weeks and five days pregnant. Her medical history included preeclampsia with severe features and HG in prior pregnancies. Initial evaluations, including CBC, complete metabolic panel, and troponin levels, were unremarkable, with normal electrolytes. However, an ECG revealed multiple arrhythmias. Cardiology and electrophysiology consultations recommended outpatient follow-up. This case highlights the serious risks HG poses to both maternal and fetal health. Although arrhythmias are a recognized complication of HG, this case is notable for their occurrence despite normal electrolyte levels, emphasizing the complex interplay between HG and cardiac function., Competing Interests: Human subjects: Consent for treatment and open access publication was obtained or waived by all participants in this study. Cape Fear Valley Health Institutional Review Board issued approval 0134-24. The GME Research Program acknowledged and logged your case report submission listed above. You may submit your abstract, poster, or manuscript for publication or presentation. No additional review is required since you have obtained the patient’s consent. Please ensure your work is approved by your advisor and/or program director to confirm that no identifiable private health information (PHI) is disclosed prior to submission for publication or presentation. Identifiable PHI may only be disclosed if it was listed in the signed consent document. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Anderson et al.)

Background/introduction: On June 24, 2022, the Supreme Court overturned the 1973 verdict in the Roe v. Wade legal suit that had established the constitutional right to medical abortion [1]. With the right to abortion now being left to the states, many restrictions regarding abortion procedures are being established [2]. We aim to evaluate whether this has led to an increase in the number of birth control prescriptions and sterilisation procedures at Cape Fear Valley Medical Centre in rural North Carolina., Methods: SlicerDicer program within EPIC medical records database was used to retrieve the records of all women that had an obstetrics and gynaecology appointment with Cape Fear Valley Health System between 11/01/21-12/31/2022 for contraception. The data was sorted into race/ethnicity, age, and types of birth control. Principal outcomes measured the trends in birth control prescription or sterilisation procedure appointments before and after the overturn of Roe v. Wade ., Results: Contraceptive pill prescriptions decreased from 51.70% to 32.40%. IUD insertions decreased from 39.40% to 11.30%. Injectable birth control remained consistent, at approximately 16%. Nexplanon implantations dropped from 20.60% to 9.70%. Lastly, sterilisation procedures increased from 6.7% to 10.7%., Conclusion: Majority of birth control prescriptions decreased following the overturn of Roe v. Wade , except for sterilisation, which increased by 4.0%. This may indicate that women are now looking for more permanent birth control options after the overturn of Roe v. Wade . Future research should investigate whether similar trends of increasing prevalence of sterilisation procedures is seen throughout the United States.

Bisphenol AF (BPAF) and TGSA are analogues of Bisphenol A (BPA). BPAF is used in polymer synthesis, while TGSA is applied in thermal papers. The EU classifies BPAF as toxic to reproduction and TGSA as a skin sensitizer. However, TGSA's other health effects remain unclear. BPAF contamination has been noted among electronic waste workers, and TGSA exposure is documented in various professions. Despite the significance of skin contact, data on skin permeation rates for BPAF and TGSA are limited. This study aimed to generate percutaneous absorption data for BPAF and TGSA following OECD guidelines. [14C]-labeled BPAF or TGSA was applied to human skin samples in vitro using Franz diffusion cells for 20 and 40 h, respectively. Key parameters such as steady-state flux, lag time, and skin permeability coefficient (K p ) were calculated. Furthermore, the distribution of the dose across different compartments, particularly within the skin, was evaluated at the conclusion of the experiment. Sequential strippings and epidermis-dermis separation were conducted for BPAF to predict the potential absorption of the remaining dose present within the skin. The permeability coefficients for BPAF and TGSA were found to be 1.9 E-03 and 1.6 E-03 cm/h, with 22% and 23% of the applied doses absorbed, respectively. Both chemicals are classified as "fast" penetrants based on their K p values. These findings suggest that BPAF and TGSA are absorbed through the skin, highlighting potential occupational risks through dermal exposure. The new percutaneous absorption data will enhance the assessment of the occupational risks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Green rooibos (Aspalathus linearis (Burm.f.) R.Dahlgren) herbal tea is popular due to its health-promoting properties. Information on its characteristic sensory profile is scarce and sensory tools to define product variation are needed. The storage conditions and time during its shelf-life are hypothesized to affect the product quality., Results: Production batches from two producers spanning 5 years (n = 57) were analyzed using descriptive sensory analysis. Primary attributes (>30 median intensity; 100% occurrence frequency) included 'hay/dried grass', 'cooked oats', 'tobacco', 'honey' and 'caramel' aromas, and astringent mouthfeel. 'Cooked vegetables', 'green grass', 'stewed fruit', 'rooibos-woody', 'marmalade' and 'cardboard' aromas, sweet taste and bitter taste were secondary attributes (10-20 median intensity; 100% occurrence frequency). The same flavor attributes were present, except for sweet-associated and fruity notes. A sensory lexicon and sensory wheels for aroma and palate attributes were constructed from the data. The shelf-life stability of green rooibos was evaluated in moisture-impermeable (pouches) and moisture-permeable (sachets) packaging at 25 and 40 °C at 60% relative humidity over 24 weeks. Green rooibos samples stored in pouches at 4 °C were also evaluated. Storage in sachets led to moisture uptake (~10 g (100 g) -1 dry basis) and an increase in water activity (>0.6), causing degradation of chlorophyll and dihydrochalcones. Changes in color and sensory profile (decreased vegetal, cereal and cardboard aromas and increased sweet-associated and fruity aromas) were evident and more pronounced at the higher storage temperature., Conclusions: Storage at ≤25 °C in moisture-impermeable packaging material is recommended for green rooibos herbal tea. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

The anti-cancer potential of Cyclopia species (honeybush) has been demonstrated in several models. The present study investigated the effects of aqueous and polyphenol-enriched (PE) extracts of C. subternata and C. genistoides , as well as mangiferin and hesperidin, on different cell growth parameters in human liver (HepG2) and colon (HT-29) cancer cells. Mangiferin and hesperidin were most abundant in C. genistoides and C. subternata , respectively. Cyclopia subternata extracts had the highest ferric-reducing antioxidant capacity. Following exposure of the cells to the extracts and compounds, cell viability, proliferation, and death (apoptosis and autophagy) were determined. Cyclopia subternata extracts reduced cell viability and inhibited cell proliferation the most, associated with depletion of ATP. In HepG2 cells, the PE extracts were less effective than the aqueous extracts in reducing cell viability but more effective in inhibiting cell proliferation. Despite disrupting cell growth, none of the extracts induced apoptosis. The aqueous extracts affected autophagy in both cancer cells. Disruption of mitochondrial membrane integrity by the different extracts, presumably via polyphenol/iron interactions, is postulated to be involved; however, mangiferin and hesperidin had no effect, suggesting that other polyphenols and/or complex interactions between compounds are likely responsible for the differential cytotoxic and/or cytoprotective effects of the extracts., Competing Interests: All authors declare that they have no conflicts of interest., (© 2024 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.)

Temperature is the most influential condition affecting insect development and population dynamics. Understanding its impact and other important factors, such as diet, could provide fine-scale predictions of species abundance and distribution in space and time. The two-spotted stink bug, Bathycoelia distincta Distant (Hemiptera: Pentatomidae), is a significant pest of macadamia in South Africa for which limited information on developmental biology exists. Here, for the first time, variations in key developmental parameters of the B. distincta biology were studied systematically. The developmental duration, survival rate, development rate, lower developmental threshold (T min ), optimum developmental threshold (T opt ), upper developmental threshold (T max ), and thermal constant were quantified for each developmental stage of B. distincta . In addition, the effect of diet (macadamia nut and sweetcorn) on the developmental duration and survival rate were quantified. This study was conducted at five constant temperatures (18, 21, 22, 25, and 29 °C) with relative humidity (RH) variations. The developmental duration from egg to adult decreased significantly with increased temperature: 21 °C (±60 days) to 29 °C (±32 days). The survival rate was significantly different for instar 2 between temperatures. Nymphs (instars 2 and 4) developed faster on the sweetcorn diet than on a macadamia diet, but the total developmental time did not differ significantly between the diets. Development from egg to adult required 783 degree days (DD), with a T min of 13.5 °C, T opt of 29.5 °C, and T max of 38 °C. Ongoing global warming will increase the population growth of B. distincta through increased development rate, resulting in more damage to macadamia nuts. Understanding the developmental biology and thresholds for the DD model of B. distincta is fundamental for predicting its phenology and outbreaks in macadamia orchards., Competing Interests: The authors declare no conflicts of interest.

Aim: This study aims to elucidate the pathology of PMFs in the South African population, establish correlations between fracture patterns and international classification guidelines and evaluate the interobserver reliability of current classifications., Methods: A retrospective review was conducted in a multicentre analysis over a one-year period from January 2019 to December 2019 at our institution. Computer tomography scans for foot and ankle injuries were reviewed, and posterior malleolus fractures were included. Pathoanatomical data was collected and analysed according to known classification systems and subsequent treatment modalities evaluated. A panel of observers individually reviewed radiographic data to determine interobserver reliability., Results: A total of 71 patients were included with a mean age of 41 ± 13.4 years (range 18-78) and a female predominant population (69%). A greater proportion of injuries were high energy (23.9%), with significant fragment comminution (53.5%), and half (52.1%) of all injuries were subluxated/dislocated at presentation. A total of 93% of injuries were managed operatively, despite theatre access limitations resulting in significant delays to fixation (19.1 days). Despite good pathoanatomical agreement with most international classifications, interobserver reliability was poor (Krippendorff α-coefficient < 0.667). Inconsistent treatment patterns in operative and non-operative strategies are reported., Conclusion: A unique patient population of younger, female individuals incurred posterior malleolar fractures due to higher energy mechanisms of injury. Whilst injury patterns were mostly comparable, significant interobserver variability was noted. Resource limitations, diagnostic challenges, poorly defined and inconsistent treatment strategies, inevitably impact outcomes within the South African population., Level of Evidence: Level III., (© 2023. The Author(s).)

The anti-allergic effects of extracts prepared from two species of honeybush, Cyclopia genistoides and Cyclopia subternata , were demonstrated in vivo in a murine allergy model for inhaled antigen induced with ovalbumin (OVA) inhalation to mimic pollen allergy. Intake of the extracts increased the production of OVA-specific immunoglobulin (Ig) E (IgE), IgG1, and IgG2a antibodies in serum and significantly suppressed anaphylactic reaction-induced body temperature decline. Moreover, the extracts significantly inhibited antigen-antibody-induced degranulation in RBL-2H3 cells. They also inhibited body temperature decline when the allergic mice were given them after antigen sensitization, indicating that anti-degranulation activity is the major mechanism underlying the anti-allergic effect of Cyclopia extracts. Despite their qualitative and quantitative differences in phenolic composition, the two extracts exhibited similar effects, suggesting that several active compounds might be involved in the activity. Therefore, oral administration of either Cyclopia extract potentially exerts a systemic anti-allergic effect, supporting the increased consumption of honeybush tea for general wellness and improved quality of life., Competing Interests: The authors have no conflicts of interest to declare., (©2024 BMFH Press.)

Phenolic compounds are known to degrade and/or undergo changes during food production and storage. Reaction kinetic modeling is generally used to define kinetic parameters of a food system and predict changes during thermal processing and storage. Data for phenolic acids and flavonoids, such as anthocyanins and flavan-3-ols, have been reviewed in detail, but the flavonoid sub-classes, dihydrochalcones and flavanones, have been mostly neglected. Other neglected phenolic classes are xanthones and benzophenones. The stability of these types of compounds is important as they are present in fruits and exposed to heat when processed into juice and jam. Other sources of the compounds are herbal teas, which are also subjected to thermal processing, either during the primary processing of the plant material, or the production of extracts for use as food ingredients. The theoretical background is given to understand the review of literature on these classes/sub-classes. Results of research on kinetic modeling are discussed in detail, while research on compound stability without the application of reaction kinetic modeling is briefly mentioned to provide context. The studies discussed included those focusing on heating during the processing and storage of model solutions, liquid foods, plant material, dried extracts, and extracts formulated with other food ingredients.

Phytosomes consist of a phytochemical bound to the hydrophilic choline head of a phospholipid. Their use in food products is gaining interest. However, literature on the use of food-grade solvents, crude plant extracts as opposed to pure compounds, and unrefined phospholipids to prepare phytosomes is limited. Furthermore, studies on compound stability are lacking. This study aimed to develop nano-phytosome vesicles prepared from inexpensive food-grade ingredients to improve the stability of polyphenolic compounds. Cyclopia subternata extract (CSE) was selected as a source of phenolic compounds. It contains substantial quantities of C -glucosyl xanthones, benzophenones, and dihydrochalcones, compounds largely neglected to date. The effect of process conditions on the complexation of CSE polyphenols with minimally refined food-grade fat-free soybean lecithin (PC) was studied. The PC:CSE ratio, sonication time, and reaction temperature were varied. This resulted in phytosomes ranging in vesicle size (113.7-312.7 nm), polydispersity index (0.31-0.48), and zeta potential (-55.0 to -38.9 mV). Variation was also observed in the yield (93.5%-96.0%), encapsulation efficiency (3.7%-79.0%), and loading capacity (LC, 1.3%-14.7%). Vesicle size and LC could be tailored by adjusting the sonication time and PC:CSE ratio, respectively. Chemical interaction between the lipid and the phenolic compounds was confirmed with nuclear magnetic resonance. Phytosomal formulation protected the compounds against degradation when freeze-dried samples were stored at 25 and 40°C for 6 months at low relative humidity. The study provided valuable information on the importance of specific process parameters in producing food-grade phytosomes with improved phenolic stability., Competing Interests: All authors declare that they have no conflicts of interest., (© 2023 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.)

The poor stability of aspalathin in aqueous solutions is a major challenge in delivering a shelf-stable ready-to-drink (RTD) green rooibos iced tea. The kinetics of aspalathin degradation and the formation of eriodictyol glucoside isomers [(S/R)-6-β-D-glucopyranosyleriodictyol and (S/R)-8-β-D-glucopyranosyleriodictyol] in aqueous buffers were modeled to understand and predict aspalathin losses during heat processing. The effects of temperature and pH on the rate constants of aspalathin degradation and eriodictyol glucoside isomer formation were determined in a 0.1 M phosphate buffer with 5.7 mM citric acid. The zero-order model best described the reaction kinetics of aspalathin degradation and eriodictyol glucoside isomer formation. Increasing the temperature and pH increased the reaction rate constants. The activation energies of the reactions were much lower at pH 6 than at pH 4, indicating that pH affected the temperature dependence of the reactions. The 8-C-glucosyl eriodictyol derivatives (RE8G and SE8G) formed at much lower rates than the 6-C-glucosyl eriodictyol derivatives (RE6G and SE6G). The metal chelators, citric acid, citrate and EDTA, drastically reduced the reaction rate constants, indicating the catalytic role of metal ions in aspalathin autoxidation. The results of the study could assist manufacturers to improve the shelf life of rooibos RTD beverages by changing the formulation and adjusting heat processing conditions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Agricultural Research Council. Published by Elsevier Ltd.. All rights reserved.)

Background: The shelf-life of a functional herbal tea-based beverage is important not only for consumer acceptability, but also for the retention of bioactive compounds. The present study aimed to clarify the role of common iced tea beverage ingredients (citric and ascorbic acids) on the shelf-life stability of an herbal tea-based beverage. A hot water extract of green Cyclopia subternata, also used as honeybush tea, was selected as the main ingredient because it provides different types of phenolic compounds associated with bioactive properties (i.e. xanthones, benzophenones, flavanones, flavones and dihydrochalcones)., Results: The model solutions were stored for 180 and 90 days at 25 and 40 °C, respectively. Changes in their volatile profiles and color were also quantified as they contribute to product quality. 3',5'-Di-β-d-glucopyranosyl-3-hydroxyphloretin (HPDG; dihydrochalcone) and, to a lesser extent, mangiferin (xanthone), were the most labile compounds. Both compounds were thus identified as critical quality indicators to determine shelf-life. The stability-enhancing activity of the acids depended on the compound; ascorbic acid and citric acid enhanced the stability of HPDG and mangiferin, respectively. However, when considering all the major phenolic compounds, the base solution without acids was the most stable. This was also observed for the color and major volatile aroma-active compounds [α-terpineol, (E)-β-damascenone, 1-p-menthen-9-al and trans-ocimenol]., Conclusion: The addition of acids, added for stability and taste in ready-to-drink iced tea beverages, could thus have unwanted consequences in that they could accelerate compositional changes and shorten the shelf-life of polyphenol-rich herbal tea beverages. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Port areas are subjected to multiple anthropic pressures that directly impact residing marine communities and deprive them of most of their essential ecological functions. Several global projects aim to rehabilitate certain ecosystem functions in port areas, such as a fish nursery function, by installing artificial fish nurseries (AFN). In theory, AFNs increase fish biodiversity and juvenile fish abundance in port areas, but studies on this subject remain scarce. Thus, the present study aimed to examine whether the use of such AFNs could restore part of the nursery function of natural habitats by increasing fish and juvenile abundance, and by decreasing predation intensity compared to bare docks. Two years of monitoring on AFNs showed they hosted 2.1 times more fish than on control docks and up to 2.4 more fish juveniles. Fish community structures were influenced by both treatment (AFN and Control) and year of monitoring. In general, AFNs hosted a greater taxonomic diversity of fish than controls. The predation intensity around these structures was significantly lower in the AFNs than in controls. Part of the definition of a fish nursery was thus verified, indicating that AFNs might be an effective restoration tool. However, we also noted that total fish abundance and Young of the Year (YOY) abundance decreased in controls, possibly due to a concentration effect. Further detailed monitoring is necessary to distinguish between these effects., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Increasingly, ecological rehabilitation is envisioned to mitigate and revert impacts of ocean sprawl on coastal marine biodiversity. While in the past studies have demonstrated the positive effects of artificial fish habitats in port areas on fish abundance and diversity, benthic colonization of these structures has not yet been taken into consideration. This could be problematic as they may provide suitable habitat for Non-Indigenous Species (NIS) and hence facilitate their spreading. The present study aimed to examine communities developing on artificial fish habitats and to observe if the number of NIS was higher than in surrounding equivalent habitats. The structures were colonized by communities that were significantly different compared to those surrounding the control habitat, and they were home to a greater number of NIS. As NIS can cause severe ecological and economical damages, our results imply that in conjunction with the ecosystem services provided by artificial fish habitats, an ecosystem disservice in the form of facilitated NIS colonization may be present. These effects have not been shown before and need to be considered to effectively decide in which situations artificial structures may be used for fish rehabilitation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Previously, we reported that a crude polyphenol-enriched fraction of Cyclopia intermedia (CPEF), a plant consumed as the herbal tea, commonly known as honeybush, reduced lipid content in 3T3-L1 adipocytes and inhibited body weight gain in obese, diabetic female leptin receptor-deficient (db/db) mice. In the current study, the mechanisms underlying decreased body weight gain in db/db mice were further elucidated using western blot analysis and in silico approaches. CPEF induced uncoupling protein 1 (UCP1, 3.4-fold, p < 0.05) and peroxisome proliferator-activated receptor alpha (PPARα, 2.6-fold, p < 0.05) expression in brown adipose tissue. In the liver, CPEF induced PPARα expression (2.2-fold, p < 0.05), which was accompanied by a 31.9% decrease in fat droplets in Hematoxylin and Eosin (H&E)-stained liver sections ( p < 0.001). Molecular docking analysis revealed that the CPEF compounds, hesperidin and neoponcirin, had the highest binding affinities for UCP1 and PPARα, respectively. This was validated with stabilising intermolecular interactions within the active sites of UCP1 and PPARα when complexed with these compounds. This study suggests that CPEF may exert its anti-obesity effects by promoting thermogenesis and fatty acid oxidation via inducing UCP1 and PPARα expression, and that hesperidin and neoponcirin may be responsible for these effects. Findings from this study could pave the way for designing target-specific anti-obesity therapeutics from C. intermedia .

Temperature tolerance is an essential component of insect fitness, and its understanding can provide a predictive framework for their distribution and abundance. The two-spotted stink bug, Bathycoelia distincta Distant, is a significant pest of macadamia. The main goal of this study was to investigate the thermal tolerance of B. distincta across different life stages. Thermal tolerance indices investigated included critical thermal maximum (CT max ), critical thermal minimum (CT min ), effects of acclimation on CT max and CT min at 20, 25, and 30 °C, and rapid heat hardening (RHH), and rapid cold hardening (RCH). The Kruskal-Wallis test was used to explore the effects of life stage and acclimation on CT max and CT min and Generalized Linear Models (GLM) for the probability of survival after pre-exposure to RHH at 41 °C for 2 h and RCH at -8 °C for 2 h. CT max and CT min varied significantly between life stages at all acclimation temperatures, but CT min (3.5 °C) varied more than CT max (2.1 °C). Higher acclimation temperatures resulted in larger variations between life stages for both CT max and CT min . A significant acclimation response was observed for the CT max of instar 2 (1.7 °C) and CT min of females (2.7 °C) across acclimation temperatures (20-30 °C). Pre-exposure significantly improved the heat and cold survival probability of instar 2 and the cold survival probability of instar 3 and males. The response between life stages was more variable in RCH than in RHH. Instar 2 appeared to be the most thermally plastic life stage of B. distincta. These results suggest that the thermal plastic traits of B. distincta life stages may enable this pest to survive in temperature regimes under the ongoing climate change, with early life stages (except for instar 2) more temperature sensitive than later life stages., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Rooibos ( Aspalathus linearis ), an indigenous South African plant and its major flavonoid, aspalathin, exhibited positive effects on glycemia and dyslipidemia in animal studies. Limited evidence exists on the effects of rooibos extract taken in combination with oral hypoglycemic and lipid-lowering medications. This study investigated the combined effects of a pharmaceutical grade aspalathin-rich green rooibos extract (GRT) with the sulfonylurea, glyburide, and atorvastatin in a type 2 diabetic ( db/db ) mouse model. Six-week-old male db/db mice and their nondiabetic lean db + littermates were divided into 8 experimental groups (n=6/group). Db/db mice were treated orally with glyburide (5 mg/kg bodyweight), atorvastatin (80 mg/kg bodyweight) and GRT (100 mg/kg bodyweight) as mono- and combination therapies respectively, for 5 weeks. An intraperitoneal glucose tolerance test was conducted at 3 weeks of treatment. Serum was collected for lipid analyses and liver tissues for histological examination and gene expression. A significant increase in the fasting plasma glucose (FPG) of the db/db mice compared to their lean counterparts (from 7.98 ± 0.83 to 26.44 ± 1.84, p < 0.0001) was observed. Atorvastatin reduced cholesterol (from 4.00 ± 0.12 to 2.93 ± 0.13, p < 0.05) and triglyceride levels (from 2.77 ± 0.50 to 1.48 ± 0.23, p < 0.05). In db/db mice, the hypotriglyceridemic effect of atorvastatin was enhanced when combined with both GRT and glyburide (from 2.77 ± 0.50 to 1.73 ± 0.35, p = 0.0002). Glyburide reduced the severity and pattern of steatotic lipid droplet accumulation from a mediovesicular type across all lobular areas, whilst combining GRT with glyburide reduced the abundance and severity of lipid droplet accumulation in the centri- and mediolobular areas. The combination of GRT, glyburide and atorvastatin reduced the abundance and severity of lipid accumulation and the intensity score compared to the administered drugs alone. The addition of either GRT or glyburide in combination with atorvastatin had no effect on blood glucose or lipid profiles, but significantly reduced lipid droplet accumulation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Patel, Muller, Joubert, Rosenkranz, Louw and Awortwe.)

Background: Temporary spanning fixation aims to provide bony stability whilst allowing access and resuscitation of the traumatised soft-tissue envelope. Conventional monolateral fixators are prone to half-pin morbidity in feet, variation in construct stability and limited weight-bearing potential. This study compares traditional delta-frame monolateral external fixators to ankle spanning circular fixators., Methods: Two cohorts were matched for demographics and fracture patterns. The quality of initial reduction and the maintenance of reduction until definitive surgery was assessed by two authors and categorised into four domains. Secondary measures included fixator costs, time to definitive surgery and complications., Results: Fifty-five delta-frames and 51 circular fixators were statistically matched for demographics and fracture pattern. "Excellent" and "Good" initial reduction was achieved in 50 (91%) delta-frames and 51 (100%) circular fixators (p = 0.027). Deterioration of initial reduction quality was seen in 12 (22%) delta-frames and two (4%) circular fixators (p < 0.001). Post-fixator dislocation occurred in five (9%) delta-frames and one (2%) circular fixator (p = 0.147). Median duration in spanned fixation was 11 days in both groups (p = 0.114). Three (5%) delta-frames and 13 (25%) circular fixators were used as definitive fixation. The mean implant cost was 4,307 USD for delta-frames and 3747 USD for circular fixators., Conclusion: Temporary spanning circular fixation offers superior intra-operative reduction and maintenance of reduction and provides more opportunity to be used as definitive fixation. Circular fixation implants also proved to be less expensive and protected against further scheduled or unscheduled returns to theatre., Level of Evidence: IV, retrospective cohort study., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Postprandial hyperglycemia is treated with the oral antidiabetic drug acarbose, an intestinal α -glucosidase inhibitor. Side effects of acarbose motivated a growing number of screening studies to identify novel α -glucosidase inhibitors derived from plant extracts and other natural sources. As "gold standard", acarbose is frequently included as the reference standard to assess the potency of these candidate α -glucosidase inhibitors, with many outperforming acarbose by several orders of magnitude. The results are subsequently used to identify suitable compounds/products with strong potential for in vivo efficacy. However, most α -glucosidase inhibitor screening studies use enzyme preparations obtained from nonmammalian sources (typically Saccharomyces cerevisiae ), despite strong evidence that inhibition data obtained using nonmammalian α -glucosidase may hold limited value in terms of identifying α -glucosidase inhibitors with actual in vivo hypoglycemic potential. The aim was to critically discuss the screening of novel α -glucosidase inhibitors from plant sources, emphasizing inconsistencies and pitfalls, specifically where acarbose was included as the reference standard. An assessment of the available literature emphasized the cruciality of stating the biological source of α -glucosidase in such screening studies to allow for unambiguous and rational interpretation of the data. The review also highlights the lack of a universally adopted screening assay for novel α -glucosidase inhibitors and the commercial availability of a standardized preparation of mammalian α -glucosidase., Competing Interests: The authors declare that they have no conflict of interest., (Thieme. All rights reserved.)

This study explored how South African food labels could be improved, to enhance customer evaluation of the overall healthiness of packaged food. Focus was given to the comparison of front-of-pack (FOP) nutrition labels as a quick assessment tool. The exploratory sequential mixed-methods design used qualitative interviews (n = 49) to gain insight into labeling challenges and select FOP nutrition labels for consumer testing. Consumers (n = 1261) randomly assessed two out of six possible FOP nutrition labels relative to a ‘no-label’ control in one of 12 online surveys, applied to a fictitious cereal product. A mixed-model analysis of variance was used to compare the differences in health ratings for the different FOP nutrition labels. The interviews revealed three themes for label improvement, that are presented over three time horizons. In terms of helping consumers identify less healthy products, the effect sizes were most prominent for health warnings (p < 0.01) and low health star ratings (p < 0.01). The findings of this research not only clarify whether FOP nutrition labeling formats used in other regions such as Europe, South America and Australia could be useful in the South African context, but they can assist policymakers and decision-makers in selecting an effective FOP label.

Stink bugs are major pests of macadamia in South Africa. Accurate identification and knowledge of species composition are important to inform management practices. The overall aims of this study were to identify stink bug species from macadamia orchards in South Africa using morphology, and to establish a DNA database based on the cytochrome c oxidase subunit 1 gene region. A total of 21 stink bug species were found in macadamia orchards in KwaZulu-Natal, Limpopo and Mpumalanga provinces. Bathycoelia distincta Distant, 1878, was the dominant species throughout all three growing regions. Two unidentified species of Boerias Kirkaldy, 1909, here designated as Boerias sp. 1 and Boerias sp. 2, were the second and third most abundant species found in KwaZulu-Natal. No species of Boerias has previously been reported in association with macadamia. Evidence of a cryptic third species of Boerias was also found. Species composition fluctuated over three growing seasons in Limpopo and differed between the three growing regions during the 2019-2020 season, highlighting the need for ongoing monitoring of these important pest species. The DNA barcode database developed in this study will be valuable for future monitoring and identifications, including cryptic or polymorphic stink bug species and different life stages.

Background: Ultrafiltration of green honeybush (Cyclopia genistoides) extract results in a by-product (retentate). Application of further separation processes for recovery of polyphenols would entail creation of additional waste. Repurposing the retentate as a food flavour ingredient provides an alternative valorization approach., Results: The retentate, suspended in water (270 g L -1 ), was heat-treated at 80 °C for 2, 4, 8 and 16 h, and at 90 °C for 2, 4, 6 and 8 h to change its sensory profile. The heat-treated retentate, diluted to beverage strength (2.15 g L -1 ), had prominent 'grape/Muscat-like' and 'marmalade/citrus' aroma and flavour notes. Overall, heating for ≤ 4 h increased the intensities of positive flavour and aroma notes, while reducing those of 'green/grass', 'hay' and bitterness, whereafter further heating only had a slight effect on the aroma profile at 80 °C (P < 0.05), but not at 90 °C (P ≥ 0.05). The heat treatments, 80 °C/4 h and 90 °C/4 h, were subsequently applied to different batches of retentate (n = 10) to accommodate the effect of natural product variation. Heating at 90 °C produced higher intensities of positive aroma attributes (P < 0.05), but was more detrimental to the phenolic stability, compared to 80 °C., Conclusion: After heat treatment, the phenolic content of C. genistoides retentate, reconstituted to beverage strength, still fell within the range of a typical 'fermented' (oxidized) honeybush leaf tea infusion. The change in phenolic composition will not diminish the benefit of an improved sensory profile for the retentate by-product through heating. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Honeybush is an indigenous herbal tea highly valued for its aroma, flavour and medicinal properties. It is protected as Geographical Indication (GI) since it is produced from a number of Cyclopia species that are endemic to South Africa. Most commonly used for honeybush tea production are C. intermedia, C. subternata and C. genistoides, differing slightly, but distinctly in flavour. Demand for species-specific honeybush tea instead of mixtures have increased, meriting a strategy for authentication of C. intermedia, C. subternata and C. genistoides. Samples of these three species were analysed, using hyperspectral imaging (HSI) in the near-infrared spectral range. The data were pre-processed and used for class-modelling, a general approach well suited for authentication purposes. Unfortunately, since the HSI data of Cyclopia species studied are very similar, the classification results obtained with individual class-models are unsatisfactory, e.g., class-models constructed for C. genistoides and C. subternata yielded correct classification rate (CCR) values of 76.4 and 83.1%, respectively. On the other hand, discriminant modelling, which is another type of classification technique, led to good classification outcomes (CCR 98.9%). However, the classical discriminant model cannot be applied for authentication purposes since it always assigns a new sample to one of the classes studied, even if in reality, it belongs to none of them. Counterfeits or non-representative samples would be incorrectly assigned by the discriminant model to one of the authentic classes. Therefore, in this study, a two-step authentication of overlapping classes is proposed, which combines the advantages of class-modelling and discriminant methods. When applied to the authentication of Cyclopia species studied, the two-step approach yielded a CCR of 97.4%, which is a significant improvement compared to results obtained with the individual class-models. The proposed approach is general and can be applied when classes studied are very similar, and individual class-models lead to unsatisfactory results., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The novel coronavirus, SARS-CoV-2, or COVID-19, has affected the world on a pandemic scale resulting in catastrophic outcomes and deaths. Currently, there is limited safety data specific to mRNA vaccine use in pregnant or lactating individuals and the potential risks to a pregnant individual and the fetus are unknown. We report an updated literature review of current information and evidence available to aid in the decision whether to vaccinate against COVID-19 currently being made by pregnant individuals and their healthcare providers so that they are able to make a well-informed recommendation and decision., (© 2021 John Wiley & Sons Ltd.)

In the pursuit of bioactive phytochemicals as a therapeutic strategy to manage metabolic risk factors for type 2 diabetes (T2D), aspalathin, C -glucosyl dihydrochalcone from rooibos ( Aspalathus linearis ), has received much attention, along with its C -glucosyl flavone derivatives and phlorizin, the apple O -glucosyl dihydrochalcone well-known for its antidiabetic properties. We provided context for dietary exposure by highlighting dietary sources, compound stability during processing, bioavailability and microbial biotransformation. The review covered the role of these compounds in attenuating insulin resistance and enhancing glucose metabolism, alleviating gut dysbiosis and associated oxidative stress and inflammation, and hyperuricemia associated with T2D, focusing largely on the literature of the past 5 years. A key focus of this review was on emerging targets in the management of T2D, as highlighted in the recent literature, including enhancing of the insulin receptor and insulin receptor substrate 1 signaling via protein tyrosine phosphatase inhibition, increasing glycolysis with suppression of gluconeogenesis by sirtuin modulation, and reducing renal glucose reabsorption via sodium-glucose co-transporter 2. We conclude that biotransformation in the gut is most likely responsible for enhancing therapeutic effects observed for the C -glycosyl parent compounds, including aspalathin, and that these compounds and their derivatives have the potential to regulate multiple factors associated with the development and progression of T2D.

Globally, the role of food labels in reducing non-communicable disease remains a point of debate. A particular area of contention is the use of health claims, an approach currently under consideration in South Africa - a developing country with vast socio-economic disparity. In the present study, in-depth interviews were conducted with 49 diverse stakeholders, including consumers and professionals from the food industry and other occupations, who shared their views about the use of health claims in a developing country context. The qualitative approach and inclusion of multiple perspectives that had not been motivated by a single stakeholder group added a novel view. Themes identified based on inductive analysis included: (i) practical barriers to label use; (ii) contextual and personal variables influencing engagement with label information; (iii) messaging preferences (for positively worded claims, compared to more cautionary statements); (iv) stakeholder complexities - mainly related to responsibility and trust; and (v) ambassadors to change. Findings indicate that there are persistent barriers to label use, such as challenges related to literacy and legibility. Furthermore, the socio-economic circumstances prevalent in South Africa drive large volumes of food sales in informal markets where labels are often not present. Unresolved questions about the substantiation and enforcement of health claims, combined with no solution being apparent for reaching consumers in the informal market, would limit the benefits that could be associated with the implementation of health claims at this point in time., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Green rooibos extract (GRE), shown to improve hyperglycemia and HDL/LDL blood cholesterol, has potential as a nutraceutical beverage ingredient. The main bioactive compound of the extract is aspalathin, a C -glucosyl dihydrochalcone. The study aimed to determine the effect of common iced tea ingredients (citric acid, ascorbic acid, and xylitol) on the stability of GRE, microencapsulated with inulin for production of a powdered beverage. The stability of the powder mixtures stored in semi-permeable (5 months) and impermeable (12 months) single-serve packaging at 30 °C and 40 °C/65% relative humidity was assessed. More pronounced clumping and darkening of the powders, in combination with higher first order reaction rate constants for dihydrochalcone degradation, indicated the negative effect of higher storage temperature and an increase in moisture content when stored in the semi-permeable packaging. These changes were further increased by the addition of crystalline ingredients, especially citric acid monohydrate. The sensory profile of the powders (reconstituted to beverage strength iced tea solutions) changed with storage from a predominant green-vegetal aroma to a fruity-sweet aroma, especially when stored at 40 °C/65% RH in the semi-permeable packaging. The change in the sensory profile of the powder mixtures could be attributed to a decrease in volatile compounds such as 2-hexenal, ( Z )-2-heptenal, ( E )-2-octenal, ( E )-2-nonenal, ( E , Z )-2,6-nonadienal and ( E )-2-decenal associated with "green-like" aromas, rather than an increase in fruity and sweet aroma-impact compounds. Green rooibos extract powders would require storage at temperatures ≤ 30 °C and protection against moisture uptake to be chemically and physically shelf-stable and maintain their sensory profiles.

Heat processing of ready-to-drink beverages is required to ensure a microbiologically safe product, however, this can result in the loss of bioactive compounds responsible for functionality. The objective of this study was to establish the thermal stability of a novel dihydrochalcone, 3',5'-di-β-d-glucopyranosyl-3-hydroxyphloretin (2), 3',5'-di-β-d-glucopyranosylphloretin (3) and other Cyclopia subternata phenolic compounds, in model solutions with or without citric acid and ascorbic acid. The solutions were heated at 93, 121 and 135 °C, relevant to pasteurisation, commercial sterilisation and ultra-high temperature (UHT) pasteurisation, respectively. For most compounds, the acids decreased the second order reaction rate constants, up to 27 times. Compound 2 (46.29 ± 0.53 (g/100 g) -1 h -1 ), and to a lesser extent compound 3 (5.94 ± 0.01 (g/100 g) -1 h -1 ) were the most thermo-unstable compounds when treated at 135 °C without added acids. Even though differential effects were observed for compounds at different temperatures and formulations, overall, the phenolic compounds were most stable under UHT pasteurisation conditions., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Purpose/Aims : To determine the incidence of corneal graft rejection in patients with approximately 1 year of steroid therapy following uDSEK. A shortened course of corticosteroids may be beneficial and a viable option in Ultrathin Descemet's Stripping Endothelial Keratoplasty (uDSEK). Materials and Methods : This retrospective cohort study evaluated corneal graft rejection rate in 170 cases of uDSEK that met inclusion criteria with approximately 1 year of steroid therapy. Corneal graft rejection was defined by anterior chamber cell/flare, keratic precipitates with/without corneal edema, and/or a rejection line. The following were collected retrospectively from January 1, 2005 through January 1, 2013: gender, age, race, indication for uDSEK, length of follow-up, length of steroid therapy. Continuous data were compared using Wilcoxon Rank Sum Test and categorical data were compared using Fisher's Exact Test. Results : The included eyes had an average length of steroid therapy of 431 days (range 85-720 days) with an average follow-up of 1274 days (range 395-3186 days). During the follow-up period without steroid, 5 of the 170 eyes were diagnosed with rejection (2.94%). Of the five eyes that experienced rejection, four eyes resolved after topical prednisone was reinstituted alone and did not require repeat uDSEK. The other eye was lost to follow-up. The cumulative probability of rejection after steroid cessation was calculated at 0.67% after 12 months, 2.20% after 18 months, 3.08% after 24 months, and 4.55% after 36 months. Conclusion : Discontinuation of topical steroid at approximately 1 year after uDSEK results in a low rate of corneal graft rejection and may prove extended use unnecessary.

Oral therapeutics used to treat type 2 diabetes and cardiovascular disease often fail to prevent the progression of disease and their comorbidities. Rooibos (Aspalathus linearis), an endemic South African plant used as an herbal tea, has demonstrated positive effects on glycemia and hypercholesterolemia. However, the treatment efficacy of rooibos extract in combination with conventional hypoglycemic and hypolipidemic medications on blood glucose and lipid profiles has not been established. This study aimed to investigate the effects of combining an aspalathin-rich green rooibos extract (Afriplex GRT™) with pioglitazone and atorvastatin, on blood glucose and lipid levels in obese diabetic (db/db) mice. Six-week-old male db/db mice and their nondiabetic lean littermate controls (db+) were divided into 8 experimental groups (n = 6/group). Db/db mice were treated daily either with pioglitazone (25 mg/kg), atorvastatin (80 mg/kg) and GRT (100 mg/kg), a combination of either drug with GRT or a combination of GRT-pioglitazone and atorvastatin for 5 weeks. Untreated vehicle controls were given dimethyl sulfoxide (0.1%) and phosphate buffered saline solution. At termination, serum and liver tissue were collected for lipid and gene expression analysis. Treatment with GRT, pioglitazone and atorvastatin combination effectively lowered fasting plasma glucose (FPG) levels in db/db mice (p = 0.02), whilst increasing body weight, liver weight, and reducing retroperitoneal fat weight. Atorvastatin monotherapy was effective at reducing cholesterol (from 4.00 ± 0.12 to 2.93 ± 0.13, p = 0.0003), LDL-C (from 0.58 ± 0.04 to 0.50 ± 0.00, p = 0.04), HDL-C (from 2.86 ± 0.05 to 2.50 ± 0.04, p = 0.0003) and TG (from 2.77 ± 0.50 to 1.48 ± 0.23, p = 0.04), compared to the untreated diabetic control. The hypotriglyceridemic effect of atorvastatin was enhanced when used in combination with both GRT and pioglitazone. The addition of pioglitazone to GRT significantly lowered FPG and TG. In db/db mice, Apoa1 was significantly downregulated in the liver, whilst Pparγ was significantly upregulated compared to their db+ counterparts. GRT monotherapy downregulated Apoa1 expression (p = 0.02). Atorvastatin combined with GRT significantly downregulated mRNA expression of Apoa1 (p = 0.03), whilst upregulating the expression of Pparγ (p = 0.03), Pparα (p = 0.002), Srebp1 (p = 0.002), and Fasn (p = 0.04). The GRT-pioglitazone-atorvastatin combination therapy downregulated Apoa1 (p = 0.006), whilst upregulating Fasn (p = 0.005), Pparα (p = 0.041), and Srebp1 (p = 0.03). Natural products can improve the efficacy of current drugs to prevent diabetes-associated complications. GRT in combination with pioglitazone enhanced the reduction of FPG, whilst the addition of atorvastatin to the combination, significantly lowered triglyceride levels. However, when GRT was used in combination with atorvastatin only cholesterol levels were affected. Although these results confirm both glucose- and lipoprotein-lowering biological effects of GRT in combination with pioglitazone and atorvastatin, increased expression of genes involved in lipogenesis, cholesterol, and fatty acid transport, β-oxidation, and synthesis and storage of fatty acids, may exacerbate the hepatotoxic effects of atorvastatin., Competing Interests: The authors have declared that no competing interests exist.

Introduction: The minor phenolic constituents of Cyclopia pubescens Eckl. & Zeyh. are unknown and one dimensional (1D) liquid chromatography (LC) is unable to provide sufficient separation., Methodology: A two-dimensional (2D) LC method incorporating normal-phasehigh performance countercurrent chromatography (NP-HPCCC) in the first dimension ( 1 D) and reversed-phase ultra-high-performance liquid chromatography (RP-UHPLC) as the second dimension ( 2 D) was developed. The analytical HPCCC method was subsequently scaled up to semi-preparative mode and fractions pooled based on phenolic sub-groups. The phenolic compounds in selected fractions were subsequently isolated using RP-HPLC on a C 18 column. Isolated compounds were identified by nuclear magnetic resonance (NMR) spectroscopy. The absolute configurations of compounds were determined by optical rotation and electronic circular dichroism spectra. Sugars were identified by gas chromatography-mass spectrometry (GC-MS) analysis., Results: The comprehensive off-line 2D CCC × LC method gave a good spread of the phenolic compounds. Orthogonality calculated using both the convex hull and conditional entropy methods were 81%. High-resolution mass spectrometric fragmentation spectra obtained from a quadrupole-time-of-flight instrument and ultraviolet-visible (UV-vis) spectral data were used to (tentatively) identify 32 phenolic compounds from the analytical CCC fractions. Of the seven isolated compounds, (2S)-5-O-[α-l-rhamnopyranosyl-(1 → 2)-β-d-glucopyranosyl]eriodictyol (3) and (2S)-5-O-[α-l-rhamnopyranosyl-(1 → 2)-β-d-glucopyranosyl]-5,7,3',4'-tetrahydroxyflavan (4) were newly identified in all plants. The other isolated compounds were identified as (2S)-5-O-[α-l-rhamnopyranosyl-(1 → 2)-β-d-glucopyranosyl]naringenin (1), R-neo-eriocitrin (2), 3-O-α-l-arabinopyranosyl-3,4-dihydroxybenzoic acid (5), 4-O-β-d-glucopyranosyl-Z-4-hydroxycinnamic acid (6) and 4-(4'-O-β-d-glucopyranosyl-4'-hydroxy-3'-methoxyphenyl)-2-butanone (7)., Conclusions: Among the 32 compounds (tentatively) identified, only six were previously identified in Cyclopia pubescens using 1D LC. Most of the isolated compounds were also identified for the first time in Cyclopia spp., improving the knowledge of the minor phenolic compounds of this genus., (© 2020 John Wiley & Sons, Ltd.)

Fractions of an ultrafiltered Cyclopia genistoides extract, respectively enriched in xanthones and benzophenones, were previously shown to inhibit mammalian α -glucosidase in vitro. The present study investigated ex vivo intestinal transport of these fractions, using excised porcine jejunal tissue, to determine whether the gut could be a predominant in vivo site of action. The major bioactive compounds, the xanthones (mangiferin, isomangiferin) and benzophenones (3- β -D-glucopyranosyliriflophenone, 3- β -D-glucopyranosyl-4- O - β -D-glucopyranosyliriflophenone) exhibited poor permeation in the absorptive direction with a relatively high efflux ratio (efflux ratio > 1). The efflux ratio of 3- β -D-glucopyranosyl-4- O - β -D-glucopyranosyliriflophenone (3.05) was similar to rhodamine 123 (2.99), a known substrate of intestinal P-glycoprotein 1 efflux transporters. Low epithelial membrane transport rates, coupled with efflux mechanisms, would effectively concentrate these bioactive compounds at the target site (gut lumen). Storage stability testing and moisture sorption assays of the xanthone-enriched fraction, benzophenone-enriched fraction, and ultrafiltered Cyclopia genistoides extract were performed to determine their susceptibility to physical and chemical degradation during storage. Hygroscopicity of the powders, indicated by moisture uptake, decreased in the order: benzophenone-enriched fraction (22.7%) > ultrafiltered Cyclopia genistoides extract (14.0%) > xanthone-enriched fraction (10.7%). 3- β -D-Glucopyranosylmaclurin, a minor benzophenone, was the least stable of the compounds, degrading faster in the benzophenone-enriched fraction than in ultrafiltered Cyclopia genistoides extract, suggesting that the ultrafiltered extract matrix may provide a degree of protection against chemical degradation. Compound degradation during 12 wk of storage at 40 °C in moisture-impermeable containers was best explained by first order reaction kinetics., Competing Interests: The authors declare that they have no conflict of interest., (Thieme. All rights reserved.)

Differential anti-proliferative and pro-apoptotic effects of aqueous extracts of green rooibos (Rg; Aspalathus linearis ) and green tea (GT; Camellia sinensis ) and an aspalathin-enriched extract of green rooibos (GRE), were investigated in primary rat hepatocytes (PH) and human liver (HepG2) and colon (HT-29) cancer cells. Rooibos flavonoids, aspalathin and luteolin, and the green tea flavanol, epigallocatechin gallate (EGCG), were included to assess their contribution relative to their extract concentrations. GRE was the most effective in reducing cell growth parameters which was associated with a high total polyphenol content and high ferric reducing potential. Differential cell responses were noticed with HepG2 cells more sensitive than PH toward the induction of apoptosis by GRE. Luteolin induced apoptosis in PH and HepG2 cells while aspalathin lacked any effect. EGCG induced apoptosis in HepG2 cells while PH were resistant. HT-29 cells were resistant to apoptosis induction by the tea and pure flavonoids. Differences existed in the individual effects of the major rooibos and GT flavonoids against cell growth parameters compared to their equivalent concentrations in the extract mixtures. Diversity of the flavonoid constituents, physicochemical properties and cellular redox status governing cell survival are likely to explain the differential cell responses.

Mitochondrial dysfunction plays a major role not only in the pathogenesis of many oxidative stress or age-related diseases such as neurodegenerative as well as mental disorders but also in normal aging. There is evidence that oxidative stress and mitochondrial dysfunction are the most upstream and common events in the pathomechanisms of neurodegeneration. Cyclopia species are endemic South African plants and some have a long tradition of use as herbal tea, known as honeybush tea. Extracts of the tea are gaining more scientific attention due to their phenolic composition. In the present study, we tested not only the in vitro mitochondria-enhancing properties of honeybush extracts under physiological conditions but also their ameliorative properties under oxidative stress situations. Hot water and ethanolic extracts of C. subternata , C. genistoides , and C. longifolia were investigated. Pretreatment of human neuroblastoma SH-SY5Y cells with honeybush extracts, at a concentration range of 0.1-1 ng/ml, had a beneficial effect on bioenergetics as it increased ATP production, respiration, and mitochondrial membrane potential (MMP) after 24 hours under physiological conditions. The aqueous extracts of C. subternata and C. genistoides , in particular, showed a protective effect by rescuing the bioenergetic and mitochondrial deficits under oxidative stress conditions (400  μ M H 2 O 2 for 3 hours). These findings indicate that honeybush extracts could constitute candidates for the prevention of oxidative stress with an impact on aging processes and age-related neurodegenerative disorders potentially leading to the development of a condition-specific nutraceutical., Competing Interests: The authors declare no conflict of interest., (Copyright © 2020 Anastasia Agapouda et al.)

Isoorientin is a natural C-glucosyl flavone that is generating a lot of interest due to its multiple pharmacological activities. Increasing experimental data have shown that the robust antioxidant and anti-inflammatory properties of isoorientin remain important in ameliorating a number of metabolic complications. In fact, plants rich in isoorientin have demonstrated strong ameliorative properties against complications such as hyperglycemia, hyperlipidemia, and insulin resistance. However, while such evidence is accumulating, it has not been reviewed to better inform on the therapeutic potential of this flavone in improving human health. This review examines and extrapolates available literature on the potential beneficial or detrimental effects associated with the use of isoorientin in mitigating metabolic diseases, with a specific focus on diabetes, obesity, and insulin resistance, including associated complications. The discussion includes effective doses in various experimental settings and proposed molecular mechanisms by which isoorientin may exert its therapeutic effects. In addition, the protective effects of extracts of a number of isoorientin-rich plants against metabolic complications will be highlighted., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A phytochemical survey to obtain bioactive natural products from Ajuga pantantha afforded five new neo-clerodane diterpenoids (1-5). The structures were established by analysis of their NMR spectroscopic data, and electronic circular dichroism calculations were applied to define their absolute configurations. Compounds 2 and 5 were found to have the property of inhibiting NO production (IC 50 values < 40 μM). Molecular docking and Western blotting were used to study the mechanism of anti-inflammatory action. Furthermore, compound 5 with the highest activity was tested for its in vivo anti-inflammatory effects using a zebrafish model., Competing Interests: Declaration of Competing Interest The authors of the present manuscript have declared that no competing interests exist., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Extracts of Cyclopia species are used as food ingredients. In vitroα-glucosidase (AG) inhibition by ultrafiltered C. genistoides extract, fractions enriched in xanthones (XEF) and benzophenones (BEF), as well as mangiferin, isomangiferin, 3-β-d-glucopyranosyliriflophenone (I3G) and 3-β-d-glucopyranosyl-4-O-β-d-glucopyranosyliriflophenone (IDG) was determined with acarbose as positive control. XEF was more potent than the extract and BEF (IC 50 = 43.3, 95.5 and 205.7 μg mL -1 , respectively). Compounds demonstrated potency in the descending order: acarbose (IC 50 = 44.3 μM) > mangiferin (102.2 μM) > isomangiferin (119.8 μM) > I3G (237.5 μM) > IDG (299.4 μM). The combination index (CI) was used to determine synergism (CI < 0.7) as demonstrated for combinations of acarbose with XEF, BEF or the respective compounds at 50% and 75% effect levels. The greatest potential acarbose dose reductions (>six-fold) across all effect levels were calculated for combinations of acarbose with mangiferin or isomangiferin, explaining the greater acarbose dose reduction potential of XEF vs. BEF. The effect of batch-to-batch variation (n = 10) of raw plant material on AG inhibition was quantified at a fixed concentration (160 μg mL -1 ). XEFs (xanthone content = 223-481 g kg -1 ) achieved AG inhibition of 63-72%, whereas BEFs (benzophenone content = 114-251 g kg -1 ) achieved AG inhibition of 26-34%, with weak linear correlation (R 2 < 0.43) between target compound content of the fractions and their achieved AG inhibition. Thus, extract fractions of C. genistoides, enriched in xanthones and benzophenones, show potential in reducing the effective dose of acarbose required to prevent postprandial hyperglycaemia.

Background: More than 80% of advanced prostate cancer (PCa) cases have bone metastasis, with a 5-year survival rate of 25%. Previously, we reported that GRT, a standardized, pharmaceutical-grade aspalathin-rich extract (12.78 g aspalathin/100 g extract), prepared from green rooibos produced from the leaves and fine stems of Aspalathus linearis, inhibits the proliferation of PCa cells, meriting this investigation to determine if GRT can suppress the migration and invasion of castration-resistant prostate cancer (CRPC) cells., Purpose: In the present study, we investigated whether GRT extract can interfere with the migration and invasion of human CRPC cells., Methods: Transwell assays were used to explore the effects of GRT on the migration and invasion of CRPC cells. Micro-Western Array (MWA) and Western blot analysis were carried out to unravel the underlying molecular mechanism(s)., Results: Treatment with 25-100 μg/ml GRT suppressed the migration and invasion of LNCaP C4-2B and 22Rv1 CRPC cells. MWA and Western blot analysis indicated that GRT treatment suppressed the protein level of yes-associated protein (YAP), macrophage stimulating 1 protein (MST1), phospho-MST1/phospho-MST2 T183/T180, and paxillin, but increased the abundance of E-cadherin. Over-expression of YAP rescued the suppressive effects of GRT on migration and invasion of CRPC cells. Treatment with the major flavonoid of GRT - the C-glucosyl dihydrochalcone, aspalathin - at a concentration of 75-100 μg/ml also reduced the migration and invasion of CRPC cells, and the inhibition was partially rescued by YAP over-expression., Conclusions: GRT treatment suppresses the migration and invasion of CRPC cells via inhibition of YAP signaling and paxillin., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. The NRF grant holder (CJF Muller) acknowledges that opinions, findings, and conclusions or recommendations expressed in any publication generated by the NRF-supported research are those of the authors, and that the NRF accepts no liability whatsoever in this regard. The funding bodies had no involvement in the study design, collection, analysis and interpretation of data, writing of the manuscript, or decision to publish the work., (Copyright © 2020. Published by Elsevier GmbH.)

Mounting evidence of the ability of aspalathin to target underlying metabolic dysfunction relevant to the development or progression of obesity and type 2 diabetes created a market for green rooibos extract as a functional food ingredient. Aspalathin is the obvious choice as a chemical marker for extract standardisation and quality control, however, often the concentration of a single constituent of a complex mixture such as a plant extract is not directly related to its bio-capacity, i.e. the level of in vitro bioactivity effected in a cell system at a fixed concentration. Three solvents (hot water and two EtOH-water mixtures), previously shown to produce bioactive green rooibos extracts, were selected for extraction of different batches of rooibos plant material (n = 10). Bio-capacity of the extracts, tested at 10 μg ml -1 , was evaluated in terms of glucose uptake by C2C12 and C3A cells and lipid accumulation in 3T3-L1 cells. The different solvents and inter-batch plant variation delivered extracts ranging in aspalathin content from 54.1 to 213.8 g kg -1 . The extracts were further characterised in terms of other major flavonoids (n = 10) and an enolic phenylpyruvic acid glucoside, using HPLC-DAD. The 80% EtOH-water extracts, with the highest mean aspalathin content (170.9 g kg -1 ), had the highest mean bio-capacity in the respective assays. Despite this, no significant (P≥ 0.05) correlation existed between aspalathin content and bio-capacity, while the orientin, isoorientin and vitexin content correlated moderately (r≥ 0.487; P < 0.05) with increased glucose uptake by C2C12 cells. Various multivariate analysis methods were then applied with Evolution Program-Partial Least Squares (EP-PLS) resulting in models with the best predictive power. These EP-PLS models, based on all quantified compounds, predicted the bio-capacity of the extracts for the respective cell types with RMSECV values ≤ 11.5, confirming that a complement of compounds, and not aspalathin content alone, is needed to predict the in vitro bio-capacity of green rooibos extracts. Additionally, the composition of hot water infusions of different production batches of green rooibos (n = 29) at 'cup-of-tea' equivalence was determined to relate dietary supplementation with the extract to intake in the form of herbal tea.

Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Professor Mehanna is a National Institute for Health Research (NIHR) Senior Investigator. The views expressed in this article are those of the author(s) and not necessarily those of the NIHR, or the Department of Health and Social Care.

The honeybush sensory wheel was revised, using a large sample set (n = 585) comprising of the major commercial Cyclopia species, i.e. C. intermedia, C. subternata and C. genistoides. Only positive and negative aroma attributes were included in the wheel. Chemicals were identified to serve as reference standards for the honeybush (Cyclopia spp.) lexicon. Similarly, chemical-based reference standards were identified for the rooibos (Aspalathus linearis) lexicon. From a comprehensive literature search and gas chromatography-mass spectrometry analyses, chemicals were screened by an expert panel for their suitability in terms of typicality of the target aroma. Each chemical was evaluated in a 'base tea' and compared to a specific 'reference tea' exhibiting a high intensity of the target aroma. A total of 30 and 44 chemicals for rooibos and honeybush, respectively, were selected for validation by a trained panel. Descriptive sensory analysis was conducted to assign typicality and intensity scores for each chemical representing a target aroma attribute. Several chemicals were identified as suitable reference standards for the following aroma notes: isophorone ('rooibos-woody'), maltyl isobutyrate ('caramel'), cis-3-hexenol ('green grass'), 4-oxoisophorone ('seaweed') and 2,4,6-trichloroanisole ('musty/mouldy') for rooibos; and 2-acetyl-5-methylfuran ('woody'), levulinic acid ('fynbos-sweet'), maltyl isobutyrate ('caramel'), and 2-acetylpyrrole ('nutty') for honeybush., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Obesity is a significant contributor to increased morbidity and premature mortality due to increasing the risk of many chronic metabolic diseases such as type 2 diabetes, cardiovascular disease and certain types of cancer. Lifestyle modifications such as energy restriction and increased physical activity are highly effective first-line treatment strategies used in the management of obesity. However, adherence to these behavioral changes is poor, with an increased reliance on synthetic drugs, which unfortunately are plagued by adverse effects. The identification of new and safer anti-obesity agents is thus of significant interest. In recent years, plants and their phenolic constituents have attracted increased attention due to their health-promoting properties. Amongst these, Cyclopia, an endemic South African plant commonly consumed as a herbal tea (honeybush), has been shown to possess modulating properties against oxidative stress, hyperglycemia, and obesity. Likewise, several studies have reported that some of the major phenolic compounds present in Cyclopia spp. exhibit anti-obesity effects, particularly by targeting adipose tissue. These phenolic compounds belong to the xanthone, flavonoid and benzophenone classes. The aim of this review is to assess the potential of Cyclopia extracts as an anti-obesity nutraceutical as underpinned by in vitro and in vivo studies and the underlying cellular mechanisms and biological pathways regulated by their phenolic compounds., (Copyright © 2019 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Variation in the bitter taste of Cyclopia genistoides (honeybush) herbal tea and reported modulation between its major xanthones, mangiferin and isomangiferin, prompted further investigation into the potential modulatory effects of honeybush phenolics. Combinations of crude benzophenone (BF)-, xanthone (XF)-, and flavanone (FF)-rich fractions and their major individual phenolic compounds were analysed by descriptive sensory analysis. The fractions were prepared from a bitter, hot water extract of green C. genistoides. Fraction BF, which is below the bitter threshold (intensity 10 on 100-point scale), enhanced the bitter intensity of XF and FF slightly (p < 0.05), although none of the major individual benzophenones retained this bitter enhancing effect. On the contrary, 3-β-d-glucopyranosyl-4-β-d-glucopyranosyloxyiriflophenone, the major benzophenone in BF, significantly (p < 0.05) decreased the bitter taste of XF, at a low concentration, whereas FF suppressed the bitter intensity of XF and mangiferin, the major xanthone present in XF. Hesperidin, however, had no effect on the bitter intensity of XF. In contrast, (2S)-5-[α-L-rhamnopyranosyl-(1→2)-β-d-glucopyranosyloxy]-naringenin, the major compound of FF, significantly (p < 0.05) enhanced the bitter taste of XF when added at concentrations comparable to that of 'fermented' honeybush tea infusions. The concentration-dependence of these bitter taste interactions may be responsible for the variable bitter intensity of C. genistoides herbal tea., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: Androgen ablation therapy is the primary treatment for metastatic prostate cancer (PCa). However, the majority of PCa patients receiving the androgen deprivation therapy develop recurrent castration-resistant prostate cancer (CRPC) within two years. Chemotherapies show little effect on prolonging survival of CRPC patients and new treatments are needed. Previous studies reported that the extracts from rooibos (Aspalathus linearis) exhibit chemopreventive properties in some cancer models, including skin, liver and oesophagus cancers in animals. We therefore investigate if extracts from rooibos can suppress the proliferation of CRPC cells., Purpose: We investigated whether an aspalathin-rich green rooibos extract (GRT™; 12.78 g aspalathin/100 g extract) demonstrates anti-cancer activity against CRPC cells., Methods: High performance liquid chromatography (HPLC) was used to profile the major flavonoids in GRT. Hoechst-dye proliferation assay, 3,4,5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide (MTT) viability assay and flow cytometry assay were used to explore the effects of GRT on the proliferation and cell cycle progression of CRPC cells. Comet assay was used to survey whether GRT induces apoptosis in CRPC cells. LNCaP 104-R1 xenograft nude mice model was used to determine the inhibitory effect of GRT on CRPC tumors in vivo. Micro-Western Array (MWA) and Western blot analysis were carried out to unravel the underlying molecular mechanism., Results: GRT contained aspalathin as the most abundant flavonoid. GRT suppressed the proliferation and survival of LNCaP 104-R1, LNCaP FGC and PC-3 PCa cells. Flow cytometry analysis showed that GRT decreased the population of PCa cells in S phase but increased the cell population in G2/M phase. Comet assay confirmed that GRT induced apoptosis in LNCaP 104-R1 cells. Gavage of 400 mg/kg GRT suppressed LNCaP 104-R1 xenografts in castrated nude mice. MWA and Western blot analysis indicated that GRT treatment suppressed Akt1, phospho-Akt Ser473, Cdc2, Bcl-2, TRAF4 and Aven, but increased activated Caspase 3, cytochrome c, and p27 Kip1 . Overexpression of Akt rescued the suppressive effects of GRT on CRPC cells. Co-treatment of GRT with Bcl-2 inhibitor ABT-737, PI3K inhibitor LY294002 and Akt inhibitor GSK 690693 exhibited additive inhibitory effect on proliferation of CRPC cells., Conclusions: GRT suppresses the proliferation of CRPC cells via inhibition of Akt signaling., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

An aspalathin-rich green rooibos extract (Afriplex GRT™) has demonstrated anti-diabetic and hypolipidemic properties, while also moderately inhibiting CYP3A4 activity, suggesting a potential for herb-drug interaction. The present study, therefore, evaluated the effects of orally administered GRT on the pharmacokinetics of atorvastatin and metformin in Wistar rats. Wistar rats were orally treated with GRT (50 mg/kg BW), atorvastatin (40 mg/kg BW) or metformin (150 mg/kg BW) alone or 50 mg/kg BW GRT in combination with 40 mg/kg BW atorvastatin or 150 mg/kg BW metformin. Blood samples were collected at 0, 10, and 30 min and 1, 2, 4, 6, and 8 h and plasma samples obtained for Liquid chromatography-mass spectrometry (LC-MS/MS) analyses. Non-compartment and two-compartment pharmacokinetic parameters of atorvastatin and metformin in the presence or absence of GRT were determined by PKSolver version 2.0 software. Membrane transporter proteins, ATP-binding cassette sub-family C member 2 ( Abcc 2), solute carrier organic anion transporter family, member 1b2 ( Slco1b2 ), ATP-binding cassette, sub-family B (MDR/TAP), member 1A ( Abcb1a ), and organic cation transporter 1 ( Oct1 ) mRNA expression were determined using real-time PCR expression data normalized to β-actin and hypoxanthine-guanine phosphoribosyltransferase (HPRT), respectively. Co-administration of GRT with atorvastatin substantially increased the maximum plasma concentration (C max ) and area of the plasma concentration-time curve (AUC 0-8 ) of atorvastatin by 5.8-fold ( p = 0.03) and 5.9-fold (p = 0.02) , respectively. GRT had no effect on the plasma levels of metformin. GRT increased Abcc2 expression and metformin downregulated Abcb1a expression while the combination of GRT with atorvastatin or metformin did not significantly alter the expression of Slco1b1 or Oct1 did not significantly alter the expression of Sclo1b2 or Oct1 . Co-administration of GRT with atorvastatin in rats may lead to higher plasma concentrations and, therefore, to an increase of the exposure to atorvastatin., (Copyright © 2019 Patel, Muller, Joubert, Rosenkranz, Taylor, Louw and Awortwe.)

Consumption of rooibos ( Aspalathus linearis ) as herbal tea is growing in popularity worldwide and its health-promoting attributes are mainly ascribed to its phenolic composition, which may be affected by the brewing conditions used. An aspect so far overlooked is the impact of cold brewing vs regular brewing and microwave boiling on the poly(phenolic) profile and in vitro antioxidant capacity of infusions prepared from red ('fermented', oxidized) and green ('unfermented', unoxidized) rooibos, the purpose of the present study. By using an untargeted metabolomics-based approach (UHPLC-QTOF mass spectrometry), 187 phenolic compounds were putatively annotated in both rooibos types, with flavonoids, tyrosols, and phenolic acids the most represented type of phenolic classes. Multivariate statistics (OPLS-DA) highlighted the phenolic classes most affected by the brewing conditions. Similar antioxidant capacities (ORAC and ABTS assays) were observed between cold- and regular-brewed green rooibos and boiled-brewed red rooibos. However, boiling green and red rooibos delivered infusions with the highest antioxidant capacities and total polyphenol content. The polyphenol content strongly correlated with the in vitro antioxidant capacities, especially for flavonoids and phenolic acids. These results contribute to a better understanding of the impact of the preparation method on the potential health benefits of rooibos tea.

Ultrafiltration of Cyclopia genistoides extract was optimised to increase its benzophenone and xanthone content as quantified using HPLC-DAD. Regenerated cellulose (RC) and polyethersulphone membranes with molecular weight cut-offs of 10 and 30 kDa were evaluated in terms of compound enrichment, permeate flux and permeate yield, using dead-end ultrafiltration. Compound enrichment was subsequently optimised using the 10 kDa RC membrane and tangential flow ultrafiltration (TFU). The effect of extract composition on compound enrichment, due to natural variation in the source material, was assessed using extracts from different batches of plant material (n = 11). Transmembrane pressure and feed flow rate affected (p < 0.05) process efficiency (mean permeate flux, compound enrichment and membrane fouling). TFU achieved ≥20% enrichment of the target compounds, proving its suitability for preparation of a nutraceutical extract of C. genistoides., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Phenolic compounds of Aspalathus linearis (rooibos) are susceptible to oxidation during "fermentation", a process characterized by the formation of a red-brown leaf color. The role of enzymes in this process is not yet understood. An experiment with dried green rooibos plant material pre-treated at 170 °C for 30 min to denature and "inactivate" endogenous enzymes was conducted to confirm the role of oxidative enzymes. The phenolic composition of "enzyme inactivated" plant material was not significantly (p ≥ .05) affected by simulated fermentation, compared to control samples, as determined using piece-wise multivariate analysis of variance for successive time intervals. This proves that rooibos enzymes participate in the oxidation of phenolic compounds during fermentation of the plant material. A kinetic modeling approach was subsequently used to establish reaction kinetic parameters for selected rooibos phenolic compounds. The degradation of aspalathin and nothofagin and formation of eriodictyol glucosides during simulated fermentation at four temperatures from 37 to 50 °C were best described by the fractional conversion model based on first-order kinetics (r 2  > 0.98), which allows for non-zero equilibrium concentrations. The extent of degradation for other compounds was too low to enable kinetic modeling. Reaction rates for the degradation/formation of phenolic compounds during fermentation followed the Arrhenius law. Less phenolic degradation (higher equilibrium concentration), but a higher reaction rate constant, was observed at higher temperatures, which could possibly be attributed to inactivation of enzymes., (Copyright © 2019 Elsevier Ltd. All rights reserved.)