The Potential Value of Debarking Water as a Source of Polyphenolic Compounds for the Specialty Chemicals Sector.

Forest-based industries produce huge quantities of bark during their primary processing activities. In Nordic pulp and paper industries, where a wet debarking process is used for bark removal, toxic debarking water and bark press water are produced as a by-product. However, polyphenols represent an important fraction of the debarking water and bark press water. These polyphenolic compounds are of commercial interest in chemical specialty sectors since polyphenols have been proven to have diverse health benefits, and after collecting them from waste sources, they can act as alternatives to oil-based chemicals. Determining the economic potential of polyphenolic compounds, identifying their molecular structure, and determining the antioxidant capacity of these compounds present in debarking water and bark can support the identification of their potential applications. The results show that water extractions from bark have a lower efficiency than (partial) alcoholic extractions. Nevertheless, a considerable amount of low-molecular polyphenolic compounds, which are of interest for high-end applications, was found in all extracts. Bark press water has a highly versatile range of polyphenolic compounds and showed some antioxidant activity, making it a great source for the collection of polyphenolic compounds, in contrast to debarking water, which had a much lower polyphenolic content and low antioxidant activity.

Antioxidant and Antimicrobial Properties of Helichrysum italicum (Roth) G. Don Hydrosol.

(1) Background: According to the emergence and spread of antibiotic resistance, there is an urge for new promising substances. The purpose of the study was to test the antioxidant, cytotoxic and antimicrobial properties of the Helichrysum italicum (Roth) G. Don essential oil (EO) and hydrosol. (2) Methods: The antioxidant potential was determined using the DPPH (2,2-diphenyl-1-picrylhydrazyl) method. The cytotoxicity for human skin and intestinal cells was tested using primary and immortalized cell line models. The minimum inhibitory concentration (MIC) of hydrosol was then determined for six bacterial strains covering four commonly reported food pathogens. Further on, the hydrosol at a concentration of 1/8 MIC was used to test the antiadhesive effect by the crystal violet (CV) staining method. (3) Results: the EO showed a 100-times higher antioxidant and 180- to 25.000-times higher cytotoxic activity, when compared to hydrosol. Nevertheless, all bacterial strains, with the exception of Staphylococcus aureus, were sensitive to hydrosol in the range of 12.5% (V/V) for Campylobacter jejuni, to MIC values of 100% (V/V) for Escherichia coli and Pseudomonas aeruginosa. The antiadhesive potential of hydrosol was also shown. (4) Conclusions: Even though hydrosols are a by-product of the EO distillation process, they possess valuable biological activities.

Helichrysum italicum (Roth) G. Don and Helichrysum arenarium (L.) Moench infusions in reversing the traits of metabolic syndrome: a double-blind randomized comparative trial.

Health impairments characteristic for metabolic syndrome such as increased body mass, a dysregulated lipid or glucose profile and elevated blood pressure can be reversed by appropriate lifestyle modifications. Supplementing the normal diet with herbal infusions is a promising strategy. We conducted a randomised double-blind comparative study in which participants with at least two traits of metabolic syndrome consumed an infusion of either Helichrysum italicum subsp. italicum (HI, n = 14) or Helichrysum arenarium (HA, n = 13) daily for 28 days. Anthropometric and biochemical parameters were measured at baseline, at the end of the intervention and after a 2-week washout period. HI infusion consumption had a beneficial effect on anthropometric traits; significant reductions in body weight, body mass index, and visceral and total body fat were observed. In the HA group, there was a greater reduction in serum glucose levels and an improvement in the lipid profile. In both groups, high LDL levels were measured at baseline, but two weeks after the intervention, in 84% of participants in the HA group and 71% in the HI group, the levels were within the reference range. Both interventions caused a decrease in HDL but also improved serum antioxidant properties. Consuming either infusion could thus be recommended as a simple, profitable habit for individuals with traits of metabolic syndrome.

Whole transcriptome expression array analysis of human colon fibroblasts culture treated with Helichrysum italicum supports its use in traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Helichrysum italicum (HI) is a Mediterranean plant with well-reported use in traditional medicine for a wide range of applications, including digestive and liver disorders, intestinal parasitic infections, wound healing, stomach ache and asthma. However, little is known about the global mechanism behind its pleiotropic activity. AIM OF THE STUDY: The aim of this study was to explain the mechanism behind the previously demonstrated effects of HI and to justify its use in traditional medicine. MATERIALS AND METHODS: A microarray-based transcriptome analysis was used to discover the global transcriptional alterations in primary colon fibroblasts after exposure to HI infusion for 6 h and 24 h. In addition, quantitative real-time PCR was used to verify the microarray results. RESULTS: Altogether we identified 217 differentially expressed genes compared to non-treated cells, and only 8 were common to both treatments. Gene ontology analysis revealed that 24 h treatment with HI infusion altered the expression of genes involved in cytoskeletal rearrangement and cell growth, whereas pathway analysis further showed the importance of interleukin signaling and transcriptional regulation by TP53. For the 6 h treatment only the process of hemostasis appeared in the results of both enrichment analyses. In functional assays, HI infusion increased cell migration and decreased blood clotting and prothrombin time. CONCLUSIONS: With the careful evaluation of the role of individual genes, especially SERPING1, ARHGAP1, IL33 and CDKN1A, represented in the enriched pathways and processes, we propose the main mode of HI action, which is wound healing. In addition to its indirect prevention of diseases resulting from the impaired barrier integrity, HI also effects inflammation and metabolic processes directly, as it regulates genes such as LRPPRC, LIPA, ABCA12, PRKAR1A and ANXA6.

Helichrysum italicum (Roth) G. Don and Helichrysum arenarium (L.) Moench Infusion Consumption Affects the Inflammatory Status and the Composition of Human Gut Microbiota in Patients with Traits of Metabolic Syndrome: A Randomized Comparative Study.

Helichrysum italicum (Roth) G. Don (HI) and Helichrysum arenarium (L.) Moench (HA) are rich in polyphenols and their infusions have beneficial effects for patients with metabolic syndrome. To investigate whether these effects are mediated by the gut microbiota, we analysed the effects of daily consumption of HI or HA infusion on the composition of gut microbiota, inflammatory status, and zonulin, a marker of gut barrier permeability. The study was a randomized, double-blind comparative trial. Thirty participants were randomly assigned to two groups and received either HA or HI tea filter bags, each containing 1 g of dried plant material, for daily consumption lasting 4 weeks. The results show that consumption of both infusions resulted in a reduction of some genera belonging to Firmicutes and in a slight but significant reduction in Shannon diversity index. Consumption of HI infusion significantly reduced serum levels of proinflammatory markers and zonulin alongside with the observed trend of Proteobacteria reduction. It can therefore be concluded that the HI and HA infusions could act as prebiotics and thus improve the intestinal environment. In addition, HI infusion has a positive impact on microbial dysbiosis and gut barrier dysfunction that occur in obesity and metabolic syndrome.

Helichrysum italicum ssp. italicum Infusion Promotes Fat Oxidation in Hepatocytes and Stimulates Energy Expenditure and Fat Oxidation after Acute Ingestion in Humans: A Pilot Study.

Helichrysum italicum is an aromatic plant with promising pharmacological activities. Bioactive compounds found in plants represent an important alternative treatment for weight loss and an infusion of H. italicum contains compounds which could have such effect. Our aim was, therefore, to investigate its acute effects on resting energy expenditure (REE) and possible differences in substrate oxidation in a pilot study. A dried, ground plant material of H. italicum ssp. italicum was infused with hot water and chemically characterized with HPLC-MS analysis. Sensory evaluation of herbal tea was performed. A randomized, crossover, controlled pilot study was then conducted on eleven healthy male subjects. The REE and substrate oxidation were measured by indirect calorimetry at baseline and 30 and 120 min after ingestion of infusion or hot water. The expression of genes involved in lipid metabolism was examined in H. italicum infusion-treated hepatocytes. Several phenolic classes were identified in the infusion, caffeoylquinic acids were the most abundant, followed by pyrones and flavonols. A single ingestion of H. italicum infusion significantly increased REE by 4% and fat oxidation by 12% compared to hot water ingestion. A significant 2-fold up-regulation of ß-oxidation-related genes in HepG2 cells, exposed to H. italicum infusion, was detected. This pilot study suggests that H. italicum infusion possesses bioactive substances with potential application in obesity prevention, which could, with additional studies, become an economically interesting novel application of the plant. Clinical trial registration number: NCT04818905.

A Review and Evaluation of the Data Supporting Internal Use of Helichrysum italicum.

Helichrysum italicum is a Mediterranean plant with various pharmacological activities. Despite extensive reports on the bioactivity of the plant, its clinically studied applications have not yet been reviewed. The aim of our study was to gather information on the internal use of H. italicum and its bioactive constituents to determine its efficacy and safety for human use. We reviewed research articles that have not been previously presented in this context and analyzed relevant clinical studies with H. italicum. Cochranelibrary.com revealed six eligible clinical trials with H. italicum that examined indications for pain management, cough, and mental exhaustion. Although the efficacy of H. italicum has been demonstrated both in in vitro tests and in humans, it is difficult to attribute results from clinical trials to H. italicum alone, as it has usually not been tested as the sole component. On the other hand, clinical trials provide positive information on the safety profile since no adverse effects have been reported. We conclude that H. italicum is safe to use internally, while new clinical studies with H. italicum as a single component are needed to prove its efficacy. Based on the recent trend in H. italicum research, further studies are to be expected.

A Comparative Study of the Antioxidative Effects of Helichrysum italicum and Helichrysum arenarium Infusions.

Helichrysum arenarium (L.) Moench (abbrev. as HA) has a long tradition in European ethnomedicine and its inflorescences are approved as a herbal medicinal product. In the Mediterranean part of Europe, Helichrysum italicum (Roth) G. Don (abbrev. as HI) is more common. Since infusions from both plants are traditionally used, we aimed to compare their antioxidative potential using in vitro assays. Two morphologically distinct HI plants, HIa and HIb, were compared to a commercially available HA product. Genetic analysis using microsatellites confirmed a clear differentiation between HI and HA and suggested that HIb was a hybrid resulting from spontaneous hybridization from unknown HI subspecies. High-performance liquid chromatography-mass spectrometry analysis showed the highest amounts of hydroxycinnamic acids and total arzanol derivatives in HIa, whereas HIb was richest in monohydroxybenzoic acids, caffeic acids, and coumarins, and HA contained the highest amounts of flavonoids, especially flavanones. HIa exhibited the highest radical scavenging activity; it was more efficient in protecting different cell lines from induced oxidative stress and in inducing oxidative stress-related genes superoxide dismutase 1, catalase, and glutathione reductase 1. The antioxidative potential of HI was not only dependent on the morphological type of the plant but also on the harvest date, revealing important information for obtaining the best possible product. Considering the superior properties of HI compared to HA, the evaluation of HI as a medicinal plant could be recommended.

Developing an Olive Biorefinery in Slovenia: Analysis of Phenolic Compounds Found in Olive Mill Pomace and Wastewater.

The valorization of olive pomace through the extraction of phenolic compounds at an industrial scale is influenced by several factors that can have a significant impact on the feasibility of this approach. These include the types and levels of phenolic compounds that are present, the impact that seasonal variation and cultivar type have on the phenolic compound content in both olive pomace and mill effluents and the technological approach used to process the olive crop. Chemical analysis of phenolic compounds was performed using an HPLC-diode-array detector (DAD)-qTOF system, resulting in the identification of 45 compounds in olive mill wastewater and pomace, where secoiridoids comprised 50-60% of the total phenolic content. This study examined three different factors that could impact the phenolic compound content of these processing streams, including cultivar types typically grown on local farms in Slovenia, the type of downstream processing used and seasonality effects. Olive crop varieties sourced from local farms showed high variability, and the highest phenolic content was associated with the local variety "Istrska Belica". During processing, the phenolic content was on average approximately 50% higher during two-phase decanting compared to three-phase decanting and the type of compound present significantly different. An investigation into the seasonal effects revealed that the phenolic content was 20% higher during the 2019 growing season compared to 2018. A larger sample size over additional growing seasons is required to fully understand the annual variation in phenolic compound content. The methods and results used in this study provide a basis for further analysis of phenolic compounds present in the European Union's olive crop processing residues and will inform techno-economic modelling for the development of olive biorefineries in Slovenia.

HPLC-DAD-ESI-QTOF-MS Determination of Bioactive Compounds and Antioxidant Activity Comparison of the Hydroalcoholic and Water Extracts from Two Helichrysum italicum Species.

Mediterranean plant Helichrysum italicum represents a rich source of versatile bioactive compounds with potential benefits for human health. Despite extensive research on the plant's active constituents, little attention has yet been paid to characterizing the relationship between its intra-specific genetic diversity and metabolite profile. The study aimed to determine metabolic profile of H. italicum ssp. italicum (HII) and ssp. tyrrhenicum (HIT) cultivated on the experimental plantation in Slovenia and to compare the chemical composition of extracts regarding the solvent extraction process. Extracts were prepared upon conventional extract preparation procedures: maceration with 50 % methanol or ethanol and cold or hot water infusion and analyzed using High Performance Liquid Chromatography-Diode Array Detection-Electrospray Ionization-Quadrupole Time-of-Flight-Mass Spectrometry (HPLC-DAD-ESI-QTOF-MS). One hundred compounds were identified in the samples, among them several isomers and derivatives were reported for the first time, while caffeoylquinic acids and pyrones were the most abundant. Semi-quantitative comparison revealed that the extraction procedure had a greater impact on the chemical profile than genetic variability. All HIT extracts showed a higher total phenolic content compared to HII, while the antioxidant potential evaluated by 1,1-diphenyl-2-picrylhydrazil test was not proportionally higher. In addition, hot water extracts proved to be comparably active as alcoholic ones, confirming high commercial potential of Helichrysum italicum as herbal functional beverages.