Antioxidant and Photoprotective Activity of Apigenin and its Potassium Salt Derivative in Human Keratinocytes and Absorption in Caco-2 Cell Monolayers.

Ultraviolet (UV) radiation, especially types A (UVA) and B (UVB), is one of the main causes of skin disorders, including photoaging and skin cancer. Ultraviolent radiation causes oxidative stress, inflammation, p53 induction, DNA damage, mutagenesis, and oxidation of various molecules such as lipids and proteins. In recent decades, the use of polyphenols as molecules with an antioxidant and anti-inflammatory capacity has increased. However, some of these compounds are poorly soluble, and information regarding their absorption and bioavailability is scarce. The main objective of this study was to compare the intestinal absorption and biological activity of apigenin and its more soluble potassium salt (apigenin-K) in terms of antioxidant and photoprotective capacity. Photoprotective effects against UVA and UVB radiation were studied in human keratinocytes, and antioxidant capacity was determined by different methods, including trolox equivalent antioxidant capacity (TEAC), ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) assays. Finally, the intestinal absorption of both apigenins was determined using an in vitro Caco-2 cell model. Apigenin showed a slightly higher antioxidant capacity in antioxidant activity assays when compared with apigenin-K. However, no significant differences were obtained for their photoprotective capacities against UVA or UVB. Results indicated that both apigenins protected cell viability in approximately 50% at 5 J/m2 of UVA and 90% at 500 J/m2 of UVB radiation. Regarding intestinal absorption, both apigenins showed similar apparent permeabilities (Papp), 1.81 × 10-5 cm/s and 1.78 × 10-5 cm/s, respectively. Taken together, these results suggest that both apigenins may be interesting candidates for the development of oral (nutraceutical) and topical photoprotective ingredients against UVA and UVB-induced skin damage, but the increased water solubility of apigenin-K makes it the best candidate for further development.

Hydroxytyrosol extracts, olive oil and walnuts as functional components in chicken sausages.

BACKGROUND: Olive oil, hydroxytyrosol and walnut can be considered ideal Mediterranean ingredients for their high polyphenolic content and healthy properties. Three extracts of hydroxytyrosol obtained using different extraction processes (HXT 1, 2, 3) (50 ppm) were evaluated for use as antioxidants in eight different chicken sausage formulas enriched in polyunsaturated fatty acids (2.5 g 100 g-1 walnut) or using extra virgin olive oil (20 g 100 g-1 ) as fat replacer. Lipid and protein oxidation, colour, emulsion stability, and the microstructure of the resulting chicken sausages were investigated and a sensory analysis was carried out. RESULTS: The sausages with HXT extracts were found to decrease lipid oxidation and to lead to the loss of thiol groups compared with control sausages. Emulsion stability (capacity to hold water and fat) was greater in the sausages containing olive oil and walnut than in control sausages. In contrast, the HXT extracts produced high emulsion instability (increasing cooking losses). Sensory analysis suggested that two of the HXT extracts studied (HXT2 and HXT3 ) were unacceptable, while the acceptability of the other was similar to that of the control products. Sausages incorporating HXT showed different structures than control samples or sausages with olive oil, related to the composition of the emulsion. CONCLUSION: These results suggest the possibility of replacing animal fat by olive oil and walnut in order to produce healthy meat products. © 2017 Society of Chemical Industry.

Effect of long-term 50 Hz magnetic field exposure on the micronucleated polychromatic erythrocytes of mice.

In recent years extremely low-frequency magnetic fields (ELF-EMF) have become widely used in human activities, leading to an increased chance of exposure to ELF-EMF. There are few reports on in vivo mammalian genotoxic effects using micronucleus (MN) assays, which generally have been used as a short-term screening system. We analyzed the possible genotoxic effect induced by long-term exposure (7, 14, 21, 28 d) of a 50 Hz ELM-MF to mice by measuring the increase in frequency of micronucleated polychromatic erythrocyte in their bone marrow (MNPCEs) and we compared it with that induced by 50 cGy of X-rays. Subsequently, we tried to reduce this chromosomal damage by administering four antioxidants substances with radioprotective capacities: dimethyl sulfoxide (DMSO), 6-n-propyl-2-thiouracil (PTU), grape-procyanidins (P) and citrus flavonoids extract (CE). The increase in micronucleated cells was higher in both physical treatments (Control < ELF-EMF (p < 0.01) 0.001)); however, the antioxidant substances only showed a genoprotective capacity against the damage induced by ionizing radiation (Ci > PTU = DMSO (p < 0.001) >P = CE (p < 0.001). The 50 Hz ELM-MF increased MNPCEs in mouse bone marrow, expressing a genotoxic capacity. Administration of antioxidant substances with radioprotective capacities known to act through the elimination of free radicals did not diminish the genotoxic effect induced by ELM-MF.

Effects of potassium apigenin and verbena extract on the wound healing process of SKH-1 mouse skin.

Tissue repair is a complex process, which may be favoured or inhibited by different factors. Potassium apigenin (AP) and other flavonoids present in verbena extract (PLX(®) ) possess powerful anti-inflammatory properties. The aim of this study was to evaluate the effects of topical treatment with AP and PLX gels on wounds inflicted on SKH-1/CRL mice. Forty-eight SKH-1 mice were used (4 groups with 12 animals each), which were subjected to wound excision with a round scalpel, 4 mm in diameter, on the dorsal skin. The animals were divided into four groups: Group I received topical applications of apigenin gel; Group II received PLX gel; Group III received vehicle gel; Group IV acted as control. Wound contraction, reepithelialisation, inflammation and neovascularisation (by means of immunohistochemical staining with anti-laminin) were recorded at study periods established at 2, 7 and 14 days. Reepithelialisation was faster in Groups I and II at 7 days (56·25% grade 3 and 43·75% grade 4) compared with the other groups. The degree of inflammation showed improvement with a tendency towards statistical significance in Groups I and II at 2 and 7 days. Anti-laminin staining was more intense in the group treated with PLX at the 2- and 7-day periods. Topical treatment with PLX gel improved the degree of reepithelialisation and inflammation, and favoured neo-vascularisation of the wounds at 2 and 7 days following surgery.

Design of Virtual Reality Exergames for Upper Limb Stroke Rehabilitation Following Iterative Design Methods: Usability Study.

BACKGROUND: Since the early 2000s, there has been a growing interest in using exercise video games (exergames) and virtual reality (VR)-based interventions as innovative methods to enhance physical rehabilitation for individuals with multiple disabilities. Over the past decade, researchers and exercise professionals have focused on developing specialized immersive exercise video games for various populations, including those who have experienced a stroke, revealing tangible benefits for upper limb rehabilitation. However, it is necessary to develop highly engaging, personalized games that can facilitate the creation of experiences aligned with the preferences, motivations, and challenges communicated by people who have had an episode of stroke. OBJECTIVE: This study seeks to explore the customization potential of an exergame for individuals who have undergone a stroke, concurrently evaluating its usability as a technological tool in the realm of physical therapy and rehabilitation. METHODS: We introduce a playtest methodology to enhance the design of a VR exergame developed using a user-centered approach for upper limb rehabilitation in stroke survivors. Over 4 playtesting sessions, stroke survivors interacted with initial game versions using VR headsets, providing essential feedback for refining game content and mechanics. Additionally, a pilot study involving 10 stroke survivors collected data through VR-related questionnaires to assess game design aspects such as mechanics, assistance, experience, motion sickness, and immersion. RESULTS: The playtest methodology was beneficial for improving the exergame to align with user needs, consistently incorporating their perspectives and achieving noteworthy results. The pilot study revealed that users had a positive response. In the first scenario, a carpenter presents a game based on the flexion-extension movement of the elbow; the second scenario includes a tejo game (a traditional Colombian throwing game) designed around game mechanics related to the flexion-extension movement of the shoulder; and in the third scenario, a farmer challenges the player to perform a movement combining elbow flexion and extension with internal and external rotation of the shoulder. These findings suggest the potential of the studied exergame as a tool for the upper limb rehabilitation of individuals who have experienced a stroke. CONCLUSIONS: The inclusion of exergames in rehabilitation for stroke-induced hemiparesis has significantly benefited the recovery process by focusing on essential shoulder and elbow movements. These interactive games play a crucial role in helping users regain mobility and restore practical use of affected limbs. They also serve as valuable data sources for researchers, improving the system's responsiveness. This iterative approach enhances game design and markedly boosts user satisfaction, suggesting exergames have promising potential as adjunctive elements in traditional therapeutic approaches.

The use of native and protonated grapefruit biomass (Citrus paradisi L.) for cadmium(II) biosorption: equilibrium and kinetic modelling.

This paper describes the use of native and protonated grapefruit biomass, a by-product of the food industry, as an effective and low-cost biosorbent for cadmium removal from aqueous solutions. The biomass composition was analysed by high-performance liquid chromatography, scanning electron microscopy coupled with energy-dispersive X-ray analysis and Fourier transform infrared spectroscopy, showing that hydroxyl and carboxylic groups were the main functional groups implicated in Cd(II) biosorption. The effect of different parameters affecting the biosorption process were studied. The optimum removal of cadmium ions was at pH 4.5. Elution of alkaline-earth ions proved to be related with cadmium uptake, aiming for an ion-exchange mechanism. Protonated biomass showed higher adsorption affinity, binding strength and irreversibility for cadmium than native grapefruit, although the optimum metal uptake and high reaction rate was for the native form of grapefruit. Biosorption experimental data fitted Freundlich > Langmuir > Temkin equilibrium adsorption models. Data for both types of biomass were better fitted by a pseudo-second-order kinetic model, with an excellent correlation between calculated and experimental values. Because of these experimental results, and taking into account that both types of biomass displayed an exothermic and spontaneous physical adsorption process, native grapefruit can be proposed in further experiments as a cheap, effective, low-cost and environmentally friendly natural sorbent for the removal of cadmium from industrial wastewater effluents, avoiding chemical pretreatment before its use.

Genoprotective Effect of Some Flavonoids against Genotoxic Damage Induced by X-rays In Vivo: Relationship between Structure and Activity.

Flavonoids constitute a group of polyphenolic compounds characterized by a common gamma-benzo- pyrone structure considered in numerous biological systems to possess antioxidant capacity. Among the different applications of flavonoids, its genoprotective capacity against damage induced by ionizing radiation stands out, which has been related to antioxidant activity and its chemical structure. In this study, we determined the frequency of appearance of micronucleus in vivo by means of the micronucleus assay. This was conducted in mice treated with different flavonoids before and after exposure to 470 mGy X-rays; thereafter, their bone marrow polychromatophilic erythrocytes were evaluated to establish the structural factors enhancing the observed genoprotective effect. Our results in vivo show that the presence of a monomeric flavan-3-ol type structure, with absence of carbonyl group in position C4 of ring C, absence of conjugation between the carbons bearing the C2 = C3 double bond and the said ring, presence of a catechol group in ring B and characteristic hydroxylation in positions 5 and 7 of ring A are the structural characteristics that determine the highest degree of genoprotection. Additionally, a certain degree of polymerization of this flavonoid monomer, but maintaining significant levels of monomers and dimers, contributes to increasing the degree of genoprotection in the animals studied at both times of their administration (before and after exposure to X-rays).

Rosemary Diterpenes and Flavanone Aglycones Provide Improved Genoprotection against UV-Induced DNA Damage in a Human Skin Cell Model.

Overexposure to solar ultraviolet (UV) radiation is the major cause of a variety of cutaneous disorders, including sunburn, photoaging, and skin cancers. UVB radiation (290-320 nm) causes multiple forms of DNA damage, p53 induction, protein and lipid oxidation, and the generation of harmful reactive oxygen species (ROS). In recent years, botanicals containing polyphenols with antioxidant and anti-inflammatory properties as skin photoprotective agents have emerged. This study evaluated the protective effects of two formulations against UVB-induced damage in a skin cell model. One of the formulations (F2) contained a combination of citrus and olive extracts and the other one (F1) also contained a rosemary extract. The antioxidant capacity of both formulations was estimated by different in vitro methods, and the cell viability, intracellular ROS generation, mitochondrial depolarization, and DNA damage were studied in UVB-irradiated human keratinocytes. Both formulations exerted photoprotective effects on skin cells and decreased mitochondrial depolarization and DNA damage. F1 which contained iridoids, rosemary diterpenes, glycosides and aglycones of citrus flavanones, and monohydroxylated flavones exhibited higher cellular photoprotective effects and mitochondrial membrane potential restoration, as well as an enhanced capacity to decrease DNA double strand breaks and the DNA damage response. In contrast, F2, which contained mostly iridoids, citrus flavanone aglycones, and mono- and dihydroxylated flavones, exhibited a higher capacity to decrease intracellular ROS generation and radical scavenging capacity related to metal ion chelation. Both formulations showed a similar capability to decrease the number of apoptotic cells upon UVB radiation. Based on our results and those of others, we postulate that the stronger capacity of F1 to protect against UVB-induced DNA damage in human keratinocytes is related to the presence of rosemary diterpenes and citrus flavanone aglycones. Nevertheless, the presence of the dihydroxylated flavones in F2 may contribute to inhibiting the generation of metal-related free radicals. To confirm the efficacy of these formulations as potential candidates for oral/topical photoprotection, human trials are required to circumvent the limitations of the cellular model.

Green Alternatives to Synthetic Antioxidants, Antimicrobials, Nitrates, and Nitrites in Clean Label Spanish Chorizo.

Natural extracts obtained from fruit and vegetable processing are important sources of phenolic compounds and nitrates, with excellent antioxidant and antimicrobial properties. The aim of this study was to characterize and determine the antioxidant and antimicrobial capacity of several natural extracts (citric (Ct), acerola (Ac), rosemary (R), paprika, garlic, oregano, beet (B), lettuce (L), arugula (A), spinach (S), chard (Ch), celery (Ce), and watercress (W)), both in vitro and applied to a cured meat product (chorizo). For that, the volatile compounds by GC-MS and microbial growth were determined. The total phenolic and nitrate contents were measured and related with their antioxidant capacity (measured by DPPH, ABTS, FRAP, and ORAC methods) and antimicrobial capacity against Clostridium perfringens growth in vitro. In order to study the antioxidant and antimicrobial activities of the extracts in food, their properties were also measured in Spanish chorizo enriched with these natural extracts. R and Ct showed the highest antioxidant capacity, however, natural nitrate sources (B, L, A, S, Ch, Ce, and W) also presented excellent antimicrobial activity against C. perfringens. The incorporation of these extracts as preservatives in Spanish chorizo also presented excellent antioxidant and antimicrobial capacities and could be an excellent strategy in order to produce clean label dry-cured meat products.

Antioxidant and Antimicrobial Activity of Rosemary, Pomegranate and Olive Extracts in Fish Patties.

Natural extracts (rich in bioactive compounds) that can be obtained from the leaves, peels and seeds, such as the studied extracts of Pomegranate (P), Rosemary (RA, Nutrox OS (NOS) and Nutrox OVS (NOVS)), and olive (Olea europaea) extracts rich in hydroxytyrosol (HYT-F from olive fruit and HYT-L from olive leaf) can act as antioxidant and antimicrobial agents in food products to replace synthetic additives. The total phenolic compounds, antioxidant capacity (measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-Azinobis (3-ethylbenzothiazolin) -6-sulphonic acid (ABTS), Ferric Reducing Antioxidant Power (FRAP), and Oxygen Radical Absorbance Capacity (ORACH)) and their antimicrobial power (using the diffusion disk method with the Escherichia Coli, Lysteria monocytogenes, and Staphilococcus Aureus strains) were measured. The results showed that all the extracts were good antioxidant and antimicrobial compounds in vitro. On the other hand, their antioxidant and antimicrobial capacity was also measured in fish products acting as preservative agents. For that, volatile fatty acid compounds were analysed by GS-MS at day 0 and 11 from elaboration, together with total vial count (TVC), total coliform count (TCC), E. Coli, and L. monocytogenes content at day 0, 4, 7 and 11 under refrigerated storage. The fish patties suffered rapid lipid oxidation and odour and flavour spoilage associated with slight rancidity. Natural extracts from pomegranate, rosemary, and hydroxytyrosol delayed the lipid oxidation, measured as volatile compounds, and the microbiological spoilage in fish patties. Addition of natural extracts to fish products contributed to extend the shelf life of fish under retail display conditions.

Radioprotective and Antimutagenic Effects of Pycnanthus angolensis Warb Seed Extract against Damage Induced by X Rays.

Although different studies have demonstrated different applications of Pycnanthus angolensis extracts in traditional African and Asian medicine, its possible antimutagenic or genoprotective capacities have never been explored. We studied these capabilities of Pycnanthus angolensis seed extract (PASE) by means of the two micronucleus assays, determining the frequency of micronucleus (MN) yield in mouse bone marrow (in vivo) and in human lymphocytes blocked by cytochalasin B (in vitro). PASE exhibited a significant genoprotective capacity (p < 0.001) against X-rays with a protection factor of 35% in both in vivo and in vitro assays. Further, its radioprotective effects were determined by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-tetrazolium bromide (MTT) cell viability test in two cell lines: one being radiosensitive (i.e., human prostate epithelium (PNT2) cells) and the other being radioresistant (i.e., B16F10 melanoma cells). In the radiosensitive cells, PASE showed a protection factor of 35.5%, thus eliminating 43.8% of X-ray-induced cell death (p < 0.001) and a dose reduction factor of 2.5. In the radioresistant cells, a protection factor of 29% (p < 0.001) with a dose reduction factor of 4 was realized. PASE elicited a greater radioprotective capacity than the substances currently used in radiation oncology and, thus, could be developed as a nutraceutical radioprotectant for workers and patients exposed to ionizing radiation.

Antioxidant and Antimicrobial Properties of Rosemary (Rosmarinus officinalis, L.): A Review.

Nowadays, there is an interest in the consumption of food without synthetic additives and rather with the use of natural preservatives. In this regard, natural extracts of the Lamiaceae family, such as rosemary, have been studied because of its bioactive properties. Several studies have reported that rosemary extracts show biological bioactivities such as hepatoprotective, antifungal, insecticide, antioxidant and antibacterial. It is well known that the biological properties in rosemary are mainly due to phenolic compounds. However, it is essential to take into account that these biological properties depend on different aspects. Their use in foods is limited because of their odour, colour and taste. For that reason, commercial methods have been developed for the preparation of odourless and colourless antioxidant compounds from rosemary. Owing to the new applications of natural extracts in preservatives, this review gives a view on the use of natural extract from rosemary in foods and its effect on preservative activities. Specifically, the relationship between the structure and activity (antimicrobial and antioxidant) of the active components in rosemary are being reviewed.

Beneficial Effects of Different Flavonoids on Vascular and Renal Function in L-NAME Hypertensive Rats.

BACKGROUND: we have evaluated the antihypertensive effect of several flavonoid extracts in a rat model of arterial hypertension caused by chronic administration (6 weeks) of the nitric oxide synthesis inhibitor, L-NAME. METHODS: Sprague Dawley rats received L-NAME alone or L-NAME plus flavonoid-rich vegetal extracts (Lemon, Grapefruit + Bitter Orange, and Cocoa) or purified flavonoids (Apigenin and Diosmin) for 6 weeks. RESULTS: L-NAME treatment resulted in a marked elevation of blood pressure, and treatment with Apigenin, Lemon Extract, and Grapefruit + Bitter Orange extracts significantly reduced the elevated blood pressure of these animals. Apigenin and some of these flavonoids also ameliorated nitric oxide-dependent and -independent aortic vasodilation and elevated nitrite urinary excretion. End-organ abnormalities such as cardiac infarcts, hyaline arteriopathy and fibrinoid necrosis in coronary arteries and aorta were improved by these treatments, reducing the end-organ vascular damage. CONCLUSIONS: the flavonoids included in this study, specially apigenin, may be used as functional food ingredients with potential therapeutic benefit in arterial hypertension.

Moderate Effect of Flavonoids on Vascular and Renal Function in Spontaneously Hypertensive Rats.

Many studies have shown that flavonoids are effective as antihypertensive drugs in arterial hypertension. In the present work, we have analyzed the effects of some flavonoid extracts in the spontaneous hypertensive rat model (SHR). An important feature of this study is that we have used a low dose, far from those that are usually applied in human therapy or experimental animals, a dose that responded to the criterion of a potential future commercial use in human subjects. Treatments were carried out for 6 and 12 weeks in two groups of SHR rats, which received apigenin, lemon extract, grapefruit + bitter orange (GBO) extracts, and cocoa extract. Captopril was used as a positive control in the SHR group treated for 6 weeks (SHR6) and Diosmin was used as the industry reference in the SHR group treated for 12 weeks (SHR12). Captopril and GBO extracts lowered the high arterial pressure of the SHR6 animals, but none of the extracts were effective in the SHR12 group. Apigenin, lemon extract (LE), GBO, and captopril also improved aortic vascular relaxation and increased plasma and urinary excretion of nitrites, but only in the SHR6 group. Kidney and urinary thiobarbituric acid reactive substances (TBARS) were also significantly reduced by GBO in the SHR6 rats. Apigenin also improved vascular relaxation in the SHR12 group and all the flavonoids studied reduced urinary thiobarbituric acid reactive substances (TBARS) excretion and proteinuria. Vascular abnormalities, such as lumen/wall ratio in heart arteries and thoracic aorta, were moderately improved by these treatments in the SHR6 group. In conclusion, the flavonoid-rich extracts included in this study, especially apigenin, LE and GBO improved vascular vasodilatory function of young adult SHRs but only the GBO-treated rats benefited from a reduction in blood pressure. These extracts may be used as functional food ingredients with a moderate therapeutic benefit, especially in the early phases of arterial hypertension.

The Effect of Consumption of Citrus Fruit and Olive Leaf Extract on Lipid Metabolism.

Citrus fruit and olive leaves are a source of bioactive compounds such as biophenols which have been shown to ameliorate obesity-related conditions through their anti-hyperlipidemic and anti-inflammatory effect, and by regulating lipoproteins and cholesterol body levels. Citrolive™ is a commercial extract which is obtained from the combination of both citrus fruit and olive leaf extracts; hence, it is hypothesised that Citrolive™ may moderate metabolic disorders that are related to obesity and their complications. Initially, an in vitro study of the inhibition of pancreatic lipase activity was made, however, no effect was found. Both preliminary and long-term evaluations of Citrolive™ on lipid metabolism were conducted in an animal model using Wistar rats. In the preliminary in vivo screening, Citrolive™ was tested on postprandial plasma triglyceride level after the administration of an oil emulsion, and a significant reduction in postprandial triacylglycerol (TAG) levels was observed. In the long-term study, Citrolive™ was administered for 60 days on Wistar rats that were fed a high-fat diet. During the study, several associated lipid metabolism indicators were analysed in blood and faeces. At the end of the experiment, the livers were removed and weighed for group comparison. Citrolive™ treatment significantly reduced the liver-to-body-weight ratio, as supported by reduced plasma transaminases compared with control, but insignificantly reduced plasma low density lipoprotein (LDL) and postprandial TAG plasma levels. In addition, faecal analysis showed that the treatment significantly increased total cholesterol excretion. On the other hand, no effect was found on faecal TAG and pancreatic lipase in vitro. In conclusion, treatment ameliorates liver inflammation symptoms that are worsened by the effects of high fat diet.

Lemon balm extract (Melissa officinalis, L.) promotes melanogenesis and prevents UVB-induced oxidative stress and DNA damage in a skin cell model.

BACKGROUND: Solar ultraviolet (UV) radiation is one of the main causes of a variety of cutaneous disorders, including photoaging and skin cancer. Its UVB component (280-315nm) leads to oxidative stress and causes inflammation, DNA damage, p53 induction and lipid and protein oxidation. Recently, an increase in the use of plant polyphenols with antioxidant and anti-inflammatory properties has emerged to protect human skin against the deleterious effects of sunlight. OBJECTIVE: This study evaluates the protective effects of lemon balm extract (LBE) (Melissa Officinalis, L) and its main phenolic compound rosmarinic acid (RA) against UVB-induced damage in human keratinocytes. METHODS: The LBE composition was determined by HPLC analysis coupled to photodiode array detector and ion trap mass spectrometry with electrospray ionization (HPLC-DAD-ESI-IT-MS/MS). Cell survival, ROS generation and DNA damage were determined upon UVB irradiation in the presence of LBE. The melanogenic capacity of LBE was also determined. RESULTS: RA and salvianolic acid derivatives were the major compounds, but caffeic acid and luteolin glucuronide were also found in LBE. LBE and RA significantly increased the survival of human keratinocytes upon UVB radiation, but LBE showed a stronger effect. LBE significantly decreased UVB-induced intracellular ROS production. Moreover, LBE reduced UV-induced DNA damage and the DNA damage response (DDR), which were measured as DNA strand breaks in the comet assay and histone H2AX activation, respectively. Finally, LBE promoted melanogenesis in the cell model. CONCLUSIONS: These results suggest that LBE may be considered as a candidate for the development of oral/topical photoprotective ingredients against UVB-induced skin damage.

Skin photoprotective and antiageing effects of a combination of rosemary (Rosmarinus officinalis) and grapefruit (Citrus paradisi) polyphenols.

BACKGROUND: Plant polyphenols have been found to be effective in preventing ultraviolet radiation (UVR)-induced skin alterations. A dietary approach based of these compounds could be a safe and effective method to provide a continuous adjunctive photoprotection measure. In a previous study, a combination of rosemary (Rosmarinus officinalis) and grapefruit (Citrus paradisi) extracts has exhibited potential photoprotective effects both in skin cell model and in a human pilot trial. OBJECTIVE: We investigated the efficacy of a combination of rosemary (R. officinalis) and grapefruit (C. paradisi) in decreasing the individual susceptibility to UVR exposure (redness and lipoperoxides) and in improving skin wrinkledness and elasticity. DESIGN: A randomised, parallel group study was carried out on 90 subjects. Furthermore, a pilot, randomised, crossover study was carried out on five subjects. Female subjects having skin phototype from I to III and showing mild to moderate chrono- or photoageing clinical signs were enrolled in both studies. Skin redness (a* value of CIELab colour space) after UVB exposure to 1 minimal erythemal dose (MED) was assessed in the pilot study, while MED, lipoperoxides (malondialdehyde) skin content, wrinkle depth (image analysis), and skin elasticity (suction and elongation method) were measured in the main study. RESULTS: Treated subjects showed a decrease of the UVB- and UVA-induced skin alterations (decreased skin redness and lipoperoxides) and an improvement of skin wrinkledness and elasticity. No differences were found between the 100 and 250 mg extracts doses, indicating a plateau effect starting from 100 mg extracts dose. Some of the positive effects were noted as short as 2 weeks of product consumption. CONCLUSIONS: The long-term oral intake of Nutroxsun™ can be considered to be a complementary nutrition strategy to avoid the negative effects of sun exposure. The putative mechanism for these effects is most likely to take place through the inhibition of UVR-induced reactive oxygen species and the concomitant inflammatory markers (lipoperoxides and cytokines) together with their direct action on intracellular signalling pathways.

Treatment of metastatic melanoma B16F10 by the flavonoids tangeretin, rutin, and diosmin.

Melanoma is one of the most frequently metastasizing malignant neoplasias. This study examines an experimental model of pulmonary metastasis and the B16F10 cell subline, highly metastatic in the lung. Antimetastatic effects of the flavonoids tangeretin, rutin, and diosmin were analyzed, and at the same time an analysis of the metastatic activity of ethanol was performed, considered to be necessary because it is used as a vehicle for administering the flavonoids. Lentini's model, which complements the macroscopic evaluation of nodule numbers by using a stereoscopic microscope and image analysis at the microscopic level, was used. The greatest reduction in the number of metastatic nodules (52%) was obtained with diosmin; similarly, the percentages of implantation, growth index, and invasion index (79.40, 67.44, and 45.23%, respectively), were all compared with those of the ethanol group, considered to be an effective control group. Rutin- and tangeretin-treated groups also showed reductions of the same index compared with the ethanol group. It would seem that structural factors would better explain these results and the antimetastatic activity of each flavonoid and the respective metabolites.

Phenolic diterpenes, flavones, and rosmarinic acid distribution during the development of leaves, flowers, stems, and roots of Rosmarinus officinalis. Antioxidant activity.

The distribution of six compounds with three different polyphenol skeletons have been studied in Rosmarinus officinalis: phenolic diterpenes (carnosic acid, carnosol, and 12-O-methylcarnosic acid), caffeoyl derivatives (rosmarinic acid), and flavones (isoscutellarein 7-O-glucoside and genkwanin), each showing a characteristic behavior and distribution during the vegetative cycle. Only in leaves were all six compounds present, and the highest accumulation rate was related with the young stages of development. Rosmarinic acid showed the highest concentrations of all the polyphenols in all organs. The distribution of this acid in leaves, flowers, and stems suggests that in the first stages of flower growth, levels were due to in situ biosynthesis, and in the last stages, the contribution of transport phenomena was increased. The antioxidant activity of six extracts with different polyphenolic composition was evaluated in aqueous and lipid systems. The results clearly suggest that rosemary extracts are excellent antioxidants in both aqueous and lipid systems.

Flavonoid distribution during the development of leaves, flowers, stems, and roots of Rosmarinus officinalis. postulation of a biosynthetic pathway.

The distribution of seven flavonoids, eriocitrin, luteolin 3'-O-beta-d-glucuronide, hesperidin, diosmin, isoscutellarein 7-O-glucoside, hispidulin 7-O-glucoside, and genkwanin, has been studied in Rosmarinus officinalis leaves, flowers, stems, and roots during plant growth. The maximum level reached by luteolin 3'-O-beta-d-glucuronide in leaves during June-August suggests the existence of a delay between the activation of the enzymes involved in the flavanone and flavone biosynthesis. The presence of hesperidin and diosmin in the vascular system is significant, and hesperidin shows even higher levels than the phenolic diterpenes and rosmarinic acid. The distribution of flavonoids observed in R. officinalis suggests a functional and structural relationship between phytoregulators and flavonoids, where flavonoids would be "protectors" of the activity of phytoregulators. A hypothesis for the general pathway of biosynthesis of these compounds in plants of the family Labiatae is proposed.