Total Triterpenes, Polyphenols, Flavonoids, and Antioxidant Activity of Bioactive Phytochemicals of Centella asiatica by Different Extraction Techniques.

Obtaining phytochemical-rich plant extracts from natural products where the active ingredients are present in comparatively low levels in the tissue matrix is the critical initial step of any chemical analysis or bioactivity testing. The plant C. asiatica is rich in various phytochemicals, the major constituents being triterpenes and flavonoids, as well as other polyphenols, leading to a number of bioactivities. In this study, an attempt was made to achieve several green technology principles, while optimizing the extraction method for the efficient extraction of bioactive compounds from C. asiatica. Soxhlet extraction (SE), ultrasound-assisted extraction (UAE) with low-frequency sonication, microwave-assisted extraction (MAE) using a closed-vessel microwave digestion system, and subcritical water extraction (SWE) in a high-pressure reactor were employed to extract the bioactive compounds. The solvent system, extraction time, and solid-to-solvent ratio were varied to optimize the extraction. UAE gave the best extraction yield, while MAE gave similar results, with a solid-to-liquid ratio of 1:25, a binary solvent system of 9:1 methanol to water (v/v), and a 20 min extraction time for the extraction of triterpenes, including madecassoside, asiaticoside, madicassic acid, and asiatic acid. Investigation of different solvent systems based on water and methanol also revealed information on the extraction behavior of total triterpene content (TTC), total polyphenolic content (TPC), total flavonoid content (TFC), and the variations in the antioxidant capacity of the extracts. In this study, it was evident that UAE and MAE offer more efficient and effective extraction of bioactive compounds in terms of extraction yield, time, and minimal solvent and energy use. Furthermore, the results showed that the different solvent ratios in the extraction mixture will affect the extraction of bioactive compounds, and a binary solvent system with a combination of methanol and water was the most efficient for the studied compounds in Centella asiatica.

The Potential Use of Propolis as an Adjunctive Therapy in Breast Cancers.

Propolis is a resinous beehive product that has a wide range of biological activities, namely antimicrobial, antioxidant, and anti-inflammatory properties. Propolis is collected by the bees from plant resin and exudates to protect hives and maintain hive homeostasis. The aim of the present systematic scoping review is to explore the potential and suitability of propolis as an adjunctive treatment in breast cancers, based on the latest available experimental evidence (2012-2021). After applying the exclusion criteria, a total of 83 research publications were identified and retrieved from Scopus, Web of Science, and Pubmed. Several relevant key themes identified from the included studies were cytotoxicity, synergistic/combination treatment, improvement in bioavailability, human clinical trials, and others. A majority of the studies identified were still in the in vitro and in vivo stages. Nonetheless, we managed to identify 4 human clinical trials that demonstrated the successful use of propolis in alleviating side effects of chemotherapy and radiotherapy while increasing the quality of life of breast cancer patients, with minimal adverse effects. In conclusion, propolis, as an adjunctive treatment, may have therapeutic benefits in alleviating symptoms related to breast cancers. However, further clinical trials, preferably with higher number of participants/subjects/patients, are urgently needed.

The Potential Use of Propolis as a Primary or an Adjunctive Therapy in Respiratory Tract-Related Diseases and Disorders: A Systematic Scoping Review.

Propolis is a resinous beehive product that is collected by the bees from plant resin and exudates, to protect and maintain hive homeostasis. Propolis has been used by humans therapeutically to treat many ailments including respiratory tract-related diseases and disorders. The aim of the present systematic scoping review is to evaluate the experimental evidence to support the use of propolis as a primary or an adjunctive therapy in respiratory tract-related diseases and disorders. After applying the exclusion criteria, 158 research publications were retrieved and identified from Scopus, Web of Science, Pubmed, and Google Scholar. The key themes of the included studies were pathogenic infection-related diseases and disorders, inflammation-related disorders, lung cancers, and adverse effects. Furthermore, the potential molecular and biochemical mechanisms of action of propolis in alleviating respiratory tract-related diseases and disorders are discussed. In conclusion, the therapeutic benefits of propolis have been demonstrated by various in vitro studies, in silico studies, animal models, and human clinical trials. Based on the weight and robustness of the available experimental and clinical evidence, propolis is effective, either as a primary or an adjunctive therapy, in treating respiratory tract-related diseases.

Ultrasound assisted extraction and quantification of targeted bioactive compounds of Centella asiatica (Gotu Kola) by UHPLC-MS/MS MRM tandem mass spectroscopy.

Centella asiatica (Gotu Kola) is a green leafy vegetable rich in phytochemicals mainly including triterpenes and caffeoylquinic acids. Fresh leaves of this plant are consumed in salads and beverages in a variety of cuisines around the world. This is a well-known functional food for its neuroprotective and cognition enhancing properties in traditional societies. HPLC-DAD at lower wavelengths commonly used to identify and quantify major triterpenes of C. asiatica extracts, but associated with few drawbacks. This paper discusses a specific, sensitive and validated method developed based on UHPLC-ESI-MS-MS-MRM tandem mass spectroscopy for targeted quantification of C. asiatica bioactive compounds. The validated method enabled a precise estimation of major triterpenes and chlorogenic acid in C. asiatica in a shorter time. The findings of this study will contribute to the information on chemotype variation of C. asiatica plant grown under unique geographical, environmental and climatic conditions in New Zealand.

Can Propolis Be a Useful Adjuvant in Brain and Neurological Disorders and Injuries? A Systematic Scoping Review of the Latest Experimental Evidence.

Propolis has been used therapeutically for centuries. In recent years, research has demonstrated its efficacy as a potential raw material for pharmaceuticals and nutraceuticals. The aim of the present scoping review is to examine the latest experimental evidence regarding the potential use of propolis in protecting the brain and treating neurological disorders and injuries. A systematic scoping review methodology was implemented. Identification of the research themes and knowledge gap was performed. After applying the exclusion criteria, a total of 66 research publications were identified and retrieved from Scopus, Web of Science, Pubmed, and Google Scholar. Several key themes where propolis is potentially useful were subsequently identified, namely detoxification, neuroinflammation, ischemia/ischemia-reperfusion injury/traumatic brain injury, Alzheimer's disease, Parkinson's disease, and epilepsy models, depression, cytotoxicity, cognitive improvement, regenerative medicine, brain infection, and adverse effects. In conclusion, propolis is shown to have protective and therapeutic benefits in alleviating symptoms of brain and neurological disorders and injuries, demonstrated by various in vitro studies, animal models, and human clinical trials. Further clinical research into this area is needed.

Antiviral, Antibacterial, Antifungal, and Antiparasitic Properties of Propolis: A Review.

Propolis is a complex phytocompound made from resinous and balsamic material harvested by bees from flowers, branches, pollen, and tree exudates. Humans have used propolis therapeutically for centuries. The aim of this article is to provide comprehensive review of the antiviral, antibacterial, antifungal, and antiparasitic properties of propolis. The mechanisms of action of propolis are discussed. There are two distinct impacts with regards to antimicrobial and anti-parasitic properties of propolis, on the pathogens and on the host. With regards to the pathogens, propolis acts by disrupting the ability of the pathogens to invade the host cells by forming a physical barrier and inhibiting enzymes and proteins needed for invasion into the host cells. Propolis also inhibits the replication process of the pathogens. Moreover, propolis inhibits the metabolic processes of the pathogens by disrupting cellular organelles and components responsible for energy production. With regard to the host, propolis functions as an immunomodulator. It upregulates the innate immunity and modulates the inflammatory signaling pathways. Propolis also helps maintain the host's cellular antioxidant status. More importantly, a small number of human clinical trials have demonstrated the efficacy and the safety of propolis as an adjuvant therapy for pathogenic infections.

Propolis in Metabolic Syndrome and Its Associated Chronic Diseases: A Narrative Review.

Propolis is a resinous product collected by bees from plants to protect and maintain the homeostasis of their hives. Propolis has been used therapeutically by humans for centuries. This review article attempts to analyze the potential use of propolis in metabolic syndrome (MetS) and its associated chronic diseases. MetS and its chronic diseases were shown to be involved in at least seven out of the top 10 causes of death in 2019. Patients with MetS are also at a heightened risk of severe morbidity and mortality in the present COVID-19 pandemic. Propolis with its antioxidant and anti-inflammatory properties is potentially useful in ameliorating the symptoms of MetS and its associated chronic diseases. The aim of this article is to provide a comprehensive review on propolis and its therapeutic benefit in MetS and its chronic diseases, with an emphasis on in vitro and in vivo studies, as well as human clinical trials. Moreover, the molecular and biochemical mechanisms of action of propolis are also discussed. Propolis inhibits the development and manifestation of MetS and its chronic diseases by inhibiting of the expression and interaction of advanced glycation end products (AGEs) and their receptors (RAGEs), inhibiting pro-inflammatory signaling cascades, and promoting the cellular antioxidant systems.

A Global Review on Short Peptides: Frontiers and Perspectives.

Peptides are fragments of proteins that carry out biological functions. They act as signaling entities via all domains of life and interfere with protein-protein interactions, which are indispensable in bio-processes. Short peptides include fundamental molecular information for a prelude to the symphony of life. They have aroused considerable interest due to their unique features and great promise in innovative bio-therapies. This work focusing on the current state-of-the-art short peptide-based therapeutical developments is the first global review written by researchers from all continents, as a celebration of 100 years of peptide therapeutics since the commencement of insulin therapy in the 1920s. Peptide "drugs" initially played only the role of hormone analogs to balance disorders. Nowadays, they achieve numerous biomedical tasks, can cross membranes, or reach intracellular targets. The role of peptides in bio-processes can hardly be mimicked by other chemical substances. The article is divided into independent sections, which are related to either the progress in short peptide-based theranostics or the problems posing challenge to bio-medicine. In particular, the SWOT analysis of short peptides, their relevance in therapies of diverse diseases, improvements in (bio)synthesis platforms, advanced nano-supramolecular technologies, aptamers, altered peptide ligands and in silico methodologies to overcome peptide limitations, modern smart bio-functional materials, vaccines, and drug/gene-targeted delivery systems are discussed.

Partial Purification and Characterization of Bioactive Peptides from Cooked New Zealand Green-Lipped Mussel (Perna canaliculus) Protein Hydrolyzates.

Proteins from fresh New Zealand green-lipped mussels were hydrolyzed for 240 min using pepsin and alcalase. The extent of the hydrolysis, antioxidant, antimicrobial, and angiotensin-converting enzyme (ACE) inhibitory activities of each protein hydrolysate were investigated. Peptides obtained from pepsin hydrolysis after 30 min, named GPH, exhibited the highest antioxidant and ACE inhibitory activity, but no antimicrobial activity. Purification of the GPH using gel-filtration chromatography revealed that the protein fraction (GPH-IV*) containing peptides with a molecular weight (MW) below 5 kDa had the strongest antioxidant and ACE inhibitory activities. Further purification was done using reverse-phase HPLC (RP-HPLC) and the only major peak obtained (GPH-IV*-P2) had the highest antioxidant and ACE inhibitory activity. From this fraction, several bioactive peptides with an MW ≈ 5 kDa were identified using LC-MS and in silico analyses. This research highlights that green-lipped mussel protein hydrolysates could be used as a good source of bioactive peptides with potential therapeutic applications.

Lipid oxidation and vitamin D3 degradation in simulated whole milk powder as influenced by processing and storage.

Vitamin D3 levels are known to sometimes decline in fortified products, which could be due to its degradation, although the exact mechanism is unknown. In this study, the influence of processing and storage conditions on lipid oxidation and vitamin D3 degradation were studied. Simulated whole milk powders with and without heat treatment were stored for 12 months at two different storage temperatures (20 °C and 40 °C). Stored samples without heat treatment showed higher lipid oxidation products analyzed by PV and TBARS values compared to those with heat treatment. Higher storage temperature also resulted in higher levels of lipid oxidation products. The concentration of vitamin D3 was also analyzed using UHPLC-MS/MS after PTAD derivatization in stored samples. An inverse relationship was observed between lipid oxidation products and vitamin D3 content. Finally, previtamin D3 and vitamin D3 oxidation products were quantified in stored samples using MRM analysis.

Identification of Vitamin D3 Oxidation Products Using High-Resolution and Tandem Mass Spectrometry.

In a successful fortification program, the stability of micronutrients added to the food is one of the most important factors. The added vitamin D3 is known to sometimes decline during storage of fortified milks, and oxidation through fatty acid lipoxidation could be suspected as the likely cause. Identification of vitamin D3 oxidation products (VDOPs) in natural foods is a challenge due to the low amount of their contents and their possible transformation to other compounds during analysis. The main objective of this study was to find a method to extract VDOPs in simulated whole milk powder and to identify these products using LTQ-ion trap, Q-Exactive Orbitrap and triple quadrupole mass spectrometry. The multistage mass spectrometry (MSn) spectra can help to propose plausible schemes for unknown compounds and their fragmentations. With the growth of combinatorial libraries, mass spectrometry (MS) has become an important analytical technique because of its speed of analysis, sensitivity, and accuracy. This study was focused on identifying the fragmentation rules for some VDOPs by incorporating MS data with in silico calculated MS fragmentation pathways. Diels-Alder derivatization was used to enhance the sensitivity and selectivity for the VDOPs' identification. Finally, the confirmed PTAD-derivatized target compounds were separated and analyzed using ESI(+)-UHPLC-MS/MS in multiple reaction monitoring (MRM) mode. Graphical Abstract ᅟ.

Degradation studies of cholecalciferol (vitamin D3) using HPLC-DAD, UHPLC-MS/MS and chemical derivatization.

In any food fortification program, the stability of added micronutrients is an important factor. Cholecalciferol or vitamin D3 is known to isomerise under various conditions, thereby making its analysis challenging. In the current study, the effects of different parameters, such as temperature, iodine, acidic conditions, and oxidation, on the isomerisation of vitamin D3 were studied using HPLC-DAD and UHPLC-MS/MS. Vitamin D3 thermally and reversibly transforms to pre-vitamin D3 type isomers. In the presence of iodine, cis/trans isomerisation of both cholecalciferol and pre-vitamin D3 takes place to form trans-vitamin D3 and tachysterol, respectively. Another isomer, isotachysterol, was formed under acidic conditions. The different rates of reaction of these products with a dienophile through the Diels-Alder reaction confirmed the formation of vitamin D3 isomerisation products. The derivatization enhanced the ionisation efficiency of vitamin D3 and its isomers in UHPLC-MS/MS and improved the separation and fragmentation enabling sensitive detection.

Structural features of a novel polysaccharide isolated from a New Zealand Maori mushroom Iliodiction cibarium.

A purified water-soluble fraction (ICP5) of a polysaccharide, isolated from a local Maori mushroom Iliodiction cibarium in New Zealand, was investigated for its structural properties. Size exclusion chromatography and dynamic light scattering showed that ICP5 had a large MW of 1.6 × 10(5) Da with a hydrodynamic diameter of 83 ± 8 nm. Particle size measurements also displayed the tendency of ICP5 to aggregate when suspended in water. The results of GC-MS, FTIR and NMR analyses allowed some characteristics of the chemical structure of ICP5 to be determined. GC-MS results showed that ICP5 contained only glucose (81.61%), galactose (12.90%) and mannose (5.49%) monomers. The characterized fragment structures of ICP5 were found to be dominantly consisting of uronic acids, which formed a backbone containing 1,4-ß-D-GlcpA. A small amount of unsaturated uronic acid also appeared to be present.

Influence of heat curing on structure and physicochemical properties of phenolic acid loaded proteinaceous electrospun fibers.

Effects of heat treatment on structure and physicochemical properties of zein (Ze) and gallic acid loaded zein (Ze-GA) electrospun fiber mats were investigated. The electrospun fiber mats displayed different surface and physicochemical properties after being heat-cured at 150 °C for 24 h, which were closely related to the initial amount of loaded gallic acid. The gallic acid was released from the Ze-GA fiber mats in a constant manner, but heat curing decreased the rate of release. Heat curing remarkably increased the molecular weight of the Ze and Ze-GA electrospun fiber mats. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) analysis of the fiber mats indicated variations in zein protein secondary structure after heat curing. (13)C solid state NMR (SS-NMR) confirmed the presence of a different chemical environment among the fiber mats. The fabrication of heat-cured zein based electrospun fibers in this study may find applications in the food packaging industry.

Chemical modification of New Zealand hoki (Macruronus novaezelandiae) skin gelatin and its properties.

Chemical modifications of gelatin from New Zealand hoki (Macruronus novaezelandiae) skins were carried out using three different cross-linking agents, namely, genipin, glutaraldehyde and caffeic acid, at different concentrations. The chemically modified gelatins exhibited better physical properties, such as higher gel strength, melting point, and rheological properties than did the uncross-linked gelatin. Gelatin cross-linked with glutaraldehyde had higher gel strength and melting point (231 g, 21.9°C) than those cross-linked with caffeic acid (229 g, 21.6°C) and genipin (211 g, 20.5°C) at concentrations of 0.133, 0.111, and 0.044M, respectively. The elastic modulus (G') and the loss modulus (G″) of chemically cross-linked gelatins were higher than those of the uncross-linked ones. These improved physicochemical properties of gelatin could lead to the development of products in the food industry that meet consumer demands.

Evaluation of gallic acid loaded zein sub-micron electrospun fibre mats as novel active packaging materials.

The applicability of gallic acid loaded zein (Ze-GA) electrospun fibre mats towards potential active food packaging material was evaluated. The surface chemistry of the electrospun fibre mats was determined using X-ray photon spectroscopy (XPS). The electrospun fibre mats showed low water activity and whitish colour. Thermogravimetric analysis (TGA) and Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy revealed the stability of the fibre mats over time. The Ze-GA fibre mats displayed similar rapid release profiles, with Ze-GA 20% exhibiting the fastest release rate in water as compared to the others. Gallic acid diffuses from the electrospun fibres in a Fickian diffusion manner and the data obtained exhibited a better fit to Higuchi model. L929 fibroblast cells were cultured on the electrospun fibres to demonstrate the absence of cytotoxicity. Overall, the Ze-GA fibre mats demonstrated antibacterial activity and properties consistent with those considered desirable for active packaging material in the food industry.

Optimizing metabolic stress disinfection and disinfestation components to control Pseudococcus longispinus.

Metabolic stress disinfection and disinfestation (MSDD) has been demonstrated to effectively control longtailed mealybug, Pseudococcus longispinus (Targioni Tozzetti). Standard components previously used for testing MSDD system included a 30-min physical phase of short cycles pressure changes followed by a 60-min chemical phase using ethanol vapor at 10 kPa. This study investigated the effect of varying the following MSDD components on mealybug mortality: duration of the physical and chemical phases, ethanol concentration, and extent of vacuum during the chemical phase. Mealybug mortality responses were analyzed, and the components were optimized using binary logistical regression to achieve 99% mortality of three life stages of the longtailed mealybug (adults, second- and third-instar nymphs and crawlers). Data indicated that the optimal components to achieve 99% mortality of all life stages were a 30-min physical phase and a 45-min chemical phase with 275 mg/liter ethanol at 30 kPa. Optimized components were obtained using binary logistical regression models. These optimized components yielded a 15-min reduction in total treatment time and a 20-kPa decrease in pressure during the chemical phase. Achievement of optimal insecticidal efficacy required all four MSDD components. Nevertheless, optimization and validation achieved 17 and 22% reductions in duration of treatment time and extent of vacuum, respectively.

Physicochemical properties of bread dough and finished bread with added pectin fiber and phenolic antioxidants.

UNLABELLED: Comparative studies were conducted in this paper to investigate the effects of added dietary fiber (DF) and/or phenolic antioxidants on the properties of bread dough and finished bread. Breads were developed in the absence (control bread), or presence of apple pectin and/or fruit phenolic extracts (treated breads), and subjected to quality evaluation (attributes including color, weight, and volume) and characterization of chemical and rheological properties. Chemical analyses revealed that breads with added phenolic extracts had greater antioxidant activity and higher extractable phenolic content, than control bread and the treated breads with added apple pectin(s). The measured antioxidant activity was mainly derived from the phenolics present in bread. Storage modulus G' (elasticity) and loss modulus G″ (viscocity) of the treated bread dough with added pectin(s) only were higher than those of control dough. The G' or G″ of the treated breads incorporated with a combination of a pectin and fruit phenolic extract depended on the type of phenolic extract (that is, apple and blackcurrant extracts behaved differently from kiwifruit extract). The G' and G″ at the final baking step were higher than those of other stages, indicating an increase in cross-linking among polymeric molecules and bread particles of high molecular weight. We conclude that the added pectin and/or phenolic extract had influenced bread dough cross-linking microstructure and bread properties through being involved in the interactions with bread components such as wheat proteins during dough development and bread baking. PRACTICAL APPLICATION: Dietary fibers and phytochemicals (including phenolic antioxidants) have long been recognized as the active nutrients responsible for the health benefits of fruit and vegetables to humans. Interest in incorporating bioactive ingredients such as dietary fiber and phenolic antioxidants into popular foods like bread has grown rapidly, due to the increased consumer health awareness. The added bioactive ingredients may or may not promote the development of bread dough. This paper reports the findings associated with the properties of the functional breads enhanced with apple pectin and apple, blackcurrant, and kiwifruit phenolic extracts. Results of this paper indicate that the success of the development of such functional breads is ultimately determined by the interactions among added bioactive ingredients and other bread components.

Properties of bread dough with added fiber polysaccharides and phenolic antioxidants: a review.

During breadmaking, different ingredients are used to ensure the development of a continuous protein network that is essential for bread quality. Interests in incorporating bioactive ingredients such as dietary fiber (DF) and phenolic antioxidants into popular foods such as bread have grown rapidly, due to the increased consumer health awareness. The added bioactive ingredients may or may not promote the protein cross-links. Appropriate cross-links among wheat proteins, fiber polysaccharides, and phenolic antioxidants could be the most critical factor for bread dough enhanced with DF and phenolic antioxidants. Such cross-links may influence the structure and properties of a bread system during baking. This article presents a brief overview of our current knowledge of the fate of the key components (wheat proteins, fibers, and phenolic antioxidants) and how they might interact during bread dough development and baking.

Vitamin D2 from irradiated mushrooms significantly increases femur bone mineral density in rats.

A method has been optimized for the conversion of ergosterol in mushrooms to vitamin D2, and the vitamin D-enriched mushrooms have been tested for bioavailability of vitamin D2 using a rat model. Femur bone mineral density (BMD) of the experimental group of animals fed with vitamin D2 (1 microg/d) obtained from irradiated mushrooms was significantly increased. Femur BMD of two groups was significantly higher. Femur BMD of the experimental group was significantly elevated compared to initial femur BMD of the study group. Data indicate that vitamin D2 from ultraviolet (UV)-irradiated mushrooms was well absorbed and metabolized in animals.