PREOPERATIVE USE OF MICRO-FLAVONOID FRACTION (MMF) AS PART OF A MULTIMODAL ANALGESIA IN ANORECTAL SURGERY: PROSPECTIVE, RANDOMIZED, PLACEBO-CONTROLLED, DOUBLE-BLIND STUDY PROTOCOL

Aim: To assess the efficiency of the micro-flavonoid fraction (MMF) administration prior to anorectal surgery with spinal anesthesia on postoperative pain severity reduction.Methods: Patients of our clinic who meet the following inclusion criteria are included: they must be diagnosed with surgical treatment of anorectal diseases. All participants are randomly divided into 2 groups: the first one gets a tablet with 1000 mg MFF (Detralex®), the second one gets a tablet containing starch per os 14 days before surgery (72 participants per arm). Then patients of each group will continue MFF administration for 30 days after surgery. Patients of both arms receive spinal anesthesia and undergo surgery. Following the procedure the primary and secondary outcomes are evaluated: frequency of the opioid drugs intake, the severity of the postoperative pain syndrome, duration and frequency of other drugs intake, readmission rate, overall quality of life, time from the procedure to returning to work and the complications rate, C-reactive protein level.Discussion: the MFF intake allows reducing the severity of the hemorrhoidal disease symptoms during conservative treatment. In addition, the Detralex® efficacy has been proven in postoperative pain reduction for patients diagnosed with varicose veins of the lower extremities. We intend to evaluate the feasibility of the preoperative MFF administration for the postoperative pain reduction and the decrease of the postoperative complications in patients with hemorrhoidal disease.

Antifungal Activity of Hyptis spicigera Methanol Leaf Extract and Flavonoid Fraction

Control of plant fungal diseases using synthetic fungicides continue to cause major problems to human health and the entire ecosystem. The aim of this research was to investigate the phytochemical and antifungal properties of Hyptis spicigera methanol leaf extract and flavonoid fraction on Aspergillus and Fusarium species, with a view to uncovering effective bio-fungicides for development as substitutes to chemical fungicides. Phytochemical screening revealed the presence of sterols and triterpenes, cardiac glycosides, flavonoids, tannins and alkaloids. The quantitative analysis showed that saponin (690 mg/g GAE), phenolics (220 mg/g GAE) and flavonoids (140 mg/g GAE) were found to be in high concentration. The antifungal effects of H. spicigera methanol extract on F. graminearum (21 mm) was significantly (p<0.05) higher than the control fungicide (Mancozeb). Similarly, the flavonoid fraction was more effective on A. parasitic (18 mm) than the control fungicide used. The extract and fraction exhibited MIC with range 3.13-12.5 mg/mL and MFC 6.25-12.5 mg/mL, indicating promising antifungal efficacies against A. flavus and F. graminearum. Our findings have revealed that H. spicigera flavonoid rich fraction has potential for development as effective bio-fungicide to control plant fungal diseases of the Aspergillus and Fusarium species.

Vitamin-Containing Antioxidant Formulation Reduces Carcinogen-Induced DNA Damage through ATR/Chk1 Signaling in Bronchial Epithelial Cells In Vitro

Lung cancer has the highest mortality rate worldwide and is often diagnosed at late stages, requiring genotoxic chemotherapy with significant side effects. Cancer prevention has become a major focus, including the use of dietary and supplemental antioxidants. Thus, we investigated the ability of an antioxidant formulation (AOX1) to reduce DNA damage in human bronchial epithelial cells (BEAS-2B) with and without the combination of apple peel flavonoid fraction (AF4), or its major constituent quercetin (Q), or Q-3-O-d-glucoside (Q3G) in vitro. To model smoke-related genotoxicity, we used cigarette-smoke hydrocarbon 4-[(acetoxymethyl)nitrosamino]-1-(3-pyridyl)-1-butanone (NNKOAc) as well as methotrexate (MTX) to induce DNA damage in BEAS-2B cells. DNA fragmentation, γ-H2AX immunofluorescence, and comet assays were used as indicators of DNA damage. Pre-exposure to AOX1 alone or in combination with AF4, Q, or Q3G before challenging with NNKOAc and MTX significantly reduced intracellular reactive oxygen species (ROS) levels and DNA damage in BEAS-2B cells. Although NNKOAc-induced DNA damage activated ATM-Rad3-related (ATR) and Chk1 kinase in BEAS-2B cells, pre-exposure of the cells with tested antioxidants prior to carcinogen challenge significantly reduced their activation and levels of γ-H2AX (p ≤ 0.05). Therefore, AOX1 alone or combined with flavonoids holds promise as a chemoprotectant by reducing ROS and DNA damage to attenuate activation of ATR kinase following carcinogen exposure.

ANTIBACTERIAL EFFECTIVITY OF FLAVONOID FRACTION OF RAMANIA LEAF EXTRACT (Bouea macrophylla Griffith) AGAINST Porphyromonas gingivalis

Background: Chronic periodontitis is a periodontal disease with 80% of all cases of periodontitis. The major causes are the accumulation of plaque and bacteria. The dominant bacteria in chronic periodontitis is Porphyromonas gingivalis. Treatment of chronic periodontitis can be done by scaling and root planing and supporting therapy by using mouthwash such as Chlorhexidine gluconate 0.2% which is the gold standard in the treatment of periodontal disease. Chlorhexidine gluconate 0.2% has disadvantages so that nowadays research on herbal plants is being done to find alternative medicines that are more effective. Ramania (Bouea machropylla Griffith) leaf contains flavonoids that have antibacterial properties. Objective: To analyze the antibacterial effectivity of the flavonoid fraction of Ramania leaf extract against Porphyromonas gingivalis that causes chronic periodontitis. Method: True experimental study and post-test with control group design consisting of 5 treatment groups, namely flavonoid fraction of ramania leaf extract with concentrations of 0.1%, 0.3%, and 0.5%, chlorhexidine gluconate 0.2% as a control positive and aquadest as a negative control. Each group was repeated 6 times. Antibacterial tests using the dilution method with inhibitory rates calculated using a UV-Vis spectrophotometer and killing rates were calculated using a Colony Counter. Results: The average difference in absorbance values obtained inhibitory rates at concentrations of 0.1%, 0.3%, and 0.5%. One Way Anova Test showed a significance value of 0,000 (p < 0.05). The average number of colonies after 24 hours incubation showed the results of a kill rates in the concentration group of 0.3%, 0.5%, and positive control. The Kruskal Wallis test showed a significance value of 0,000 (p < 0.05). Conclusion: The minimum inhibitory concentration (MIC) was obtained at a concentration of 0.1% and the minimum bactericidal concentration (MBC) was obtained at 0.3% concentration. Keywords: Flavonoid Fraction, MIC, MBC, Porphyromonas Gingivalis, Ramania Leaf Extract.

A Validated Ultrasound-Assisted Extraction Coupled with SPE-HPLC-DAD for the Determination of Flavonoids in By-Products of Plant Origin: An Application Study for the Valorization of the Walnut Septum Membrane

Walnut byproducts have been shown to exert functional properties, but the literature on their bioactive content is still scarce. Among walnut byproducts, walnut septum is a dry ligneous diaphragm tissue that divides the two halves of the kernel, exhibiting nutritional and medicinal properties. These functional properties are owing to its flavonoid content, and in order to explore the flavonoid fraction, an ultrasound-assisted (UAE) protocol was combined with solid phase extraction (SPE) and coupled to high-performance liquid chromatography with diode array detection (HPLC-DAD) for the determination of flavonoids in Greek walnut septa membranes belonging to Chandler, Vina, and Franquette varieties. The proposed UAE-SPE-HPLC-DAD method was validated and the relative standard deviations (RSD%) of the within-day and between-day assays were lower than 6.2 and 8.5, respectively, showing good precision, and high accuracy ranging from 90.8 (apigenin) to 97.5% (catechin) for within-day assay, and from 88.5 (myricetin) to 96.2% (catechin) for between-day assay. Overall, seven flavonoids were determined (catechin, rutin, myricetin, luteolin, quercetin, apigenin, and kaempferol) suggesting that the walnut septum is a rich source of bioactive constituents. The quantification results were further processed using ANOVA analysis to examine if there are statistically significant differences between the concentration of each flavonoid and the variety of the walnut septum.

The effect of chayote leaves (Sechium edule)’s flavonoid fraction on the reduction of the serum uric acid levels through the inhibition of xanthine oxidase activity

Uric acid is the final product of purine metabolism and is categorized as hyperuricemia when it reaches >6.0 mg.dL-1 for women and >7.0 mg.dL-1 for men. The chayote leaves (Sechium edule) contain a high amount of flavonoid and might be used as an alternative to reduce hyperuricemia. The purpose of this study is to analyze the effect of chayote leaves (Sechium edule)’s flavonoid fraction on the level of uric acid and the activity of xanthine oxidase (XO) in Sprague Dawley Rats. The flavonoid fraction (FF) was obtained by extracting the chayote leaves, fractionating with n-hexane, hydrolyzing with HCl, and finally re-fractionating with ethyl acetate. Thirty male Sprague Dawley rats were induced for hyperuricemia by potassium oxonate and broth block for 21 days, and the interventions were given orally for 14 days. The rats were divided randomly into five groups: normal control (K-), hyperuricemia control (K+), hyperuricemia with FF dose 50 mg.200g-1 body weight (P1), hyperuricemia with FF dose 100 mg.200g-1 body weight (P2) and hyperuricemia with allopurinol 1.8 mg.200g-1 body weight. Xanthine oxidase activity was measured by CheKineTM Xanthine Oxidase Assay Kit, with simple colorimetry methods. The statistical analysis for XO activity was done using Kruskal-Wallis followed by Mann Whitney. The results showed that chayote leaves (Sechium edule)’s flavonoid fraction contains apigenin, apigenin o-glucoside, and luteolin. It also has antioxidant activity with 98.45% inhibition. There was a significant reduction in xanthine oxidase activity in groups treated with FF (p <0.005). The best dose of FF affecting XO activity was 100 mg.200g-1 body weight. The combination of FF and allopurinol can be more effective in decreasing uric acid levels by inhibiting XO activity.

Multidisciplinary analysis of Italian Alpine wildflower honey reveals criticalities, diversity and value

Wildflower honeys produced in mountain grasslands are an expression of the biodiversity of these fragile habitats. Despite its importance, the botanical origin of honey is often defined without performing formal analysis. The aim of the study was to characterize six wildflower mountain honeys produced in the Italian Alps with different analytic techniques (SPME–GC–MS, HPLC-Orbitrap, cicatrizing and antioxidant activity) alongside melissopalynological analysis and botanical definition of the production area. Even though the apiaries were in mountain grasslands rich in Alpine herbaceous species, the honey could be defined as rhododendron/raspberry unifloral or raspberry and rhododendron bifloral while the honey produced at the lowest altitude differed due to the presence of linden, heather and chestnut. The non-compliance of the honey could be due to habitat (meadows and pastures) fragmentation, but also to specific compounds involved in the plant–insect relationship, such as kynurenic acid, present in a high quantity in the sample rich in chestnut pollen. 255 volatile compounds were detected as well as some well-known markers of specific botanic essences, in particular chestnut, linden and heather, also responsible for most of the differences in aroma profiling. A high correlation between nicotinaldehyde content and percentage of raspberry pollen (r = 0.853, p < 0.05) was found. Phenolic acid and hydroxy-fatty acid were predominant in the chestnut pollen dominant honey, which presented the highest antioxidant activity and the lowest cicatrizing activity, while the flavonoid fraction was accentuated in one sample (rhododendron pollen prevalent), that was also the one with the highest effect on wound closure, although all samples had similar cicatrizing effects apart from the chestnut pollen dominant honey (lowest cicatrizing activity). Our study highlighted the difficulty of producing mountain wildflower honey and the importance of a thorough characterization of this product, also to encourage its production and valorisation.

The choice of excipients in the development of a soft dosage form of phleboprotective action containing a micronized flavonoid fraction

Varicose veins are called one of the diseases of the 21st century, caused by a sedentary lifestyle and genetics. The purpose of this study was the development of ointment compositions containing the micronized flavonoid fraction and the study of the degree of release of the micronized fraction by the flavonoid from the ointment compositions with penetrating components. With the obtained ointment compositions, a complex of biopharmaceutical studies in vitro was carried out: release from ointment compositions by diffusion into a gelatin gel and dialysis through a semipermeable membrane, while the quality indicators of the ointment compositions were established. The conducted in vitro biopharmaceutical studies helped to determine the final composition of the ointment in relation to the auxiliary composition: active ingredient — 2%, PEG 1500:PEG 400 (7:3), propylene glycol — 1%.

Flavonoid fraction from chayote (Sechium edule (Jacq.) Sw) leaves reduced malondialdehyde (MDA) and tumor necrosis factor-α (TNF-α) in hyperuricemic rats

Purpose This paper aims to study the effect of the flavonoid fraction of chayote (Sechium edule (Jacq.) Sw) leaves (FFCL) on uric acid (UA) levels, oxidative stress and inflammatory markers in hyperuricemia rats. Design/methodology/approach In total, 30 Sprague–Dawley rats were divided randomly into 5 groups. A healthy control group was established. Hyperuricemia was induced by the administration of block broth and potassium oxonate for three weeks. FFCL at dosages of 50 and 100 mg/200 g BW/d or allopurinol at a dosage of 1.8 mg/200 g BW/d was given orally for 2 weeks. Statistical analysis was conducted to evaluate differences among groups before and after the intervention. Findings Treatment with two different doses FFCL (50 and 100 mg/200 g BW/d) and one dose of allopurinol (1.8 mg/200 g BW/d) for 2 weeks significantly reduced UA from 8.04 ± 0.23 to 3.88 ± 0.10; 8.03 ± 0.18 to 2.87 ± 0.10; 8.23 ± 0.21 to 2.53 ± 0.19 (p < 0.05), respectively. The oxidative stress marker malondialdehyde levels were reduced (p = 0.001) from 9.68 ± 0.28 to 4.06 ± 0.58; 10.01 ± 0.23 to 2.12 ± 0.09; 9.88 ± 0.21 to 2.02 ± 0.17 (p = 0.001). The inflammatory marker tumor necrosis factor-α (TNF-α) levels were also reduced from 26.43 ± 0.87 to 12.20 ± 0.32; 27.38 ± 0.53 to 9.60 ± 0.53; 27.55 ± 0.68 to 8.83 ± 0.21 with p = 0.001. The 100 mg/200 g BW/d FFCL decreased UA levels, oxidative stress and inflammatory markers more extensively compared to 50 mg/200 g BW/d FFCL. Research limitations/implications This study includes some limitations that may affect the generalizability of its findings. First, the flavonoid levels of FFCL were not measured. Second, other oxidative stress biomarkers (e.g. superoxide dismutase) and inflammatory biomarkers (e.g. IL-6) were not investigated. Finally, the experiments were conducted on the model animals over a relatively short period of time. Further research is needed to evaluate the effect in humans at chronic use. Practical implications Chayote (Sechium edule (Jacq.) Sw) leaves are rich in flavonoids, especially apigenin and luteolin, which can improve oxidative stress and inflammation conditions caused by hyperuricemia. Social implications Hyperuricemia is a risk factor for non-communicable diseases, mostly caused by oxidative stress and inflammation in the body due to high levels of UA, one of the treatment strategies is through diet modification. Originality/value The results of this investigation imply that the administration of the flavonoid fraction of chayote leaves has significant effects on UA and oxidative stress and inflammatory markers. Further research is necessary to confirm the results.