Comparative evaluation of hesperetin-loaded graphene oxide nanosheets (Hsp-GO) as a drug delivery system for colon cancer: synthesis and anticancer efficiency assessment.

BACKGROUND: Graphene oxide nanosheets (GONS) are recognized for their role in enhancing drug delivery and effectiveness in cancer treatment. With colon cancer being a prevalent global issue and the significant side effects associated with chemotherapy, the primary treatment for colon cancer alongside surgery, there is a critical need for novel therapeutic strategies to support patients in combating this disease. Hesperetin (HSP), a natural compound found in specific fruits, exhibits anti-cancer properties. The aim of this study is to investigate the effect of GONS on the LS174t colon cancer cell line. METHODS: In this study, an anti-cancer nano-drug was synthesized by creating a hesperetin-graphene oxide nanocomposite (Hsp-GO), which was subsequently evaluated for its efficacy through in vitro cell toxicity assays. Three systems were investigated: HSP, GONS, and HSP-loaded GONS, to determine their cytotoxic and pro-apoptotic impacts on the LS174t colon cancer cell line, along with assessing the expression of BAX and BCL2. The morphology and properties of both GO and Hsp-GO were examined using scanning electron microscopy (SEM), X-ray diffraction, and Fourier transform infrared spectroscopy (FTIR). RESULTS: The Hsp-GO nanocomposite displayed potent cytotoxic and pro-apoptotic effects on LS174t colon cancer cells, outperforming individual treatments with HSP or GONS. Cell viability assays showed a significant decrease in cell viability with Hsp-GO treatment. Analysis of BAX and BCL2 expression revealed elevated BAX and reduced BCL2 levels in Hsp-GO treated cells, indicating enhanced apoptotic activity. Morphological analysis confirmed successful Hsp-GO synthesis, while structural integrity was supported by X-ray diffraction and FTIR analyses. CONCLUSIONS: These study highlight the potential of Hsp-GO as a promising anti-cancer nano-drug for colon cancer therapy.

Enhanced oral delivery of hesperidin-loaded sulfobutylether-ß-cyclodextrin/chitosan nanoparticles for augmenting its hypoglycemic activity: in vitro-in vivo assessment study.

Hesperidin (Hsd), a bioactive phytomedicine, experienced an antidiabetic activity versus both Type 1 and Type 2 Diabetes mellitus. However, its intrinsic poor solubility and bioavailability is a key challenging obstacle reflecting its oral delivery. From such perspective, the purpose of the current study was to prepare and evaluate Hsd-loaded sulfobutylether-ß-cyclodextrin/chitosan nanoparticles (Hsd/CD/CS NPs) for improving the hypoglycemic activity of the orally administered Hsd. Hsd was first complexed with sulfobutylether-ß-cyclodextrin (SBE-ß-CD) and the complex (CX) was found to be formed with percent complexation efficiency and percent process efficiency of 50.53 ± 1.46 and 84.52 ± 3.16%, respectively. Also, solid state characterization of the complex ensured the inclusion of Hsd inside the cavity of SBE-ß-CD. Then, Hsd/CD/CS NPs were prepared using the ionic gelation technique. The prepared NPs were fully characterized to select the most promising one (F1) with a homogenous particle size of 455.7 ± 9.04 nm, a positive zeta potential of + 32.28 ± 1.12 mV, and an entrapment efficiency of 77.46 ± 0.39%. The optimal formula (F1) was subjected to further investigation of in vitro release, ex vivo intestinal permeation, stability, cytotoxicity, and in vivo hypoglycemic activity. The results of the release and permeation studies of F1 manifested a modulated pattern between Hsd and CX. The preferential stability of F1 was observed at 4 ± 1 °C. Also, the biocompatibility of F1 with oral epithelial cell line (OEC) was retained up to a concentration of 100 µg/mL. After oral administration of F1, a noteworthy synergistic hypoglycemic effect was recorded with decreased blood glucose level until the end of the experiment. In conclusion, Hsd/CD/CS NPs could be regarded as a hopeful oral delivery system of Hsd with enhanced antidiabetic activity.

Assessment of Various Food Proteins as Structural Materials for Delivery of Hydrophobic Polyphenols Using a Novel Co-Precipitation Method.

In this study, sodium caseinate (NaCas), soy protein isolate (SPI), and whey protein isolate (WPI) were used as structural materials for the delivery of rutin, naringenin, curcumin, hesperidin, and catechin. For each polyphenol, the protein solution was brought to alkaline pH, and then the polyphenol and trehalose (as a cryo-protectant) were added. The mixtures were later acidified, and the co-precipitated products were lyophilized. Regardless of the type of protein used, the co-precipitation method exhibited relatively high entrapment efficiency and loading capacity for all five polyphenols. Several structural changes were seen in the scanning electron micrographs of all polyphenol-protein co-precipitates. This included a significant decrease in the crystallinity of the polyphenols, which was confirmed by X-ray diffraction analysis, where amorphous structures of rutin, naringenin, curcumin, hesperidin, and catechin were revealed after the treatment. Both the dispersibility and solubility of the lyophilized powders in water were improved dramatically (in some cases, >10-fold) after the treatment, with further improvements observed in these properties for the powders containing trehalose. Depending on the chemical structure and hydrophobicity of the tested polyphenols, there were differences observed in the degree and extent of the effect of the protein on different properties of the polyphenols. Overall, the findings of this study demonstrated that NaCas, WPI, and SPI can be used for the development of an efficient delivery system for hydrophobic polyphenols, which in turn can be incorporated into various functional foods or used as supplements in the nutraceutical industry.

Fine characterization and microbiota assessment as keys to understanding the positive effect of standardized natural citrus extract on broiler chickens.

The objective of this study was to investigate the effect and composition of a standardized natural citrus extract (SNCE) on both broiler chickens' growth performances and intestinal microbiota. A total of 930 one-day-old males were randomly assigned to three dietary treatments: a control treatment (CTL) in which broiler chickens were fed with a standard diet and two citrus treatments in which broiler chickens were fed with the same standard diet supplemented with 250 ppm and 2,500 ppm of SNCE, respectively. Each dietary treatment was composed of 10 experimental units (pen) of 31 broiler chickens each. Growth performances such as feed consumption, body weight, and feed conversion ratio (FCR) were recorded weekly until day 42. Litter quality was also weekly recorded while mortality was daily recorded. One broiler chicken was randomly selected from each pen (10 chickens/group) and ceca samples were collected for microbiota analysis at day 7 and 42. Chromatographic methods were used to determine molecules that enter into the composition of the SNCE. Results from the characterization of SNCE allowed to identify pectic oligosaccharides (POS) as a major component of the SNCE. In addition, 35 secondary metabolites, including eriocitrin, hesperidin, and naringin, were identified. The experiment performed on broiler chickens showed that the final body weight of broiler chickens fed diets supplemented with SNCE was higher than those fed the CTL diets (P < 0.01). Broiler cecal microbiota was impacted by age (P < 0.01) but not by the dietary supplementation of SNCE. Results indicate that SNCE allowed enhancing chickens' performances without any modulation of the cecal microbiota of broiler chickens. The characterization of SNCE allowed to identify compounds such as eriocitrin, naringin, hesperidin, and POS. Thus, opening new horizons for a better understanding of the observed effect on broiler chickens' growth performances.

Citrus extracts are increasingly being used in animal nutrition to enhance animal growth performances. Most of the available studies indicate an effect of these extracts on microbiota. However, citrus extracts can vary a lot. Indeed, the composition of citrus extract depends on parameters such as the citrus species, the extraction methods, and the inclusion rate. This variation is very important to take into consideration before using a citrus extract. The objective here was to evaluate a commercially available standardized natural citrus extract in terms of composition and effect on broiler chickens' performances and microbiota. Results showed that standardized natural citrus extract positively affects the final weight of broilers, but no effect was observed on chickens' caecal microbiota. The characterization of the standardized natural citrus extract reveals pectic oligosaccharides as major compounds as well as 35 others molecules. Most of these compounds are well described for their beneficial effect on animals' performances and health. In conclusion, the standardized natural citrus extract showed beneficial effects on broilers' performances. These effects are not correlated with broilers microbiota modulation and may be explained by the composition of the product.

Dynamic assessment of the relationship between oxidative stress and apoptotic pathway in embryonic fibroblast cells exposed to glycidamide: possible protective role of hesperidin.

Worldwide research is being conducted to determine the level of acrylamide (ACR) that humans are exposed to from food and environmental sources. Glycidamide (GA) is an important epoxide metabolite of ACR, and its cytotoxicity is stronger than ACR. In this study, it was aimed to elucidate the effects and underlying mechanisms of GA on the induction of apoptosis in embryonic fibroblast cells. The toxicogenomic profile of GA was studied in terms of both apoptotic and oxidative stress. Embryonic fibroblast cells were exposed to GA (1 and 1000 µM) in the presence and absence of hesperidin (Hes) (20 µM) or vitamin C (VitC) (50 µM) for 24 h. Cell viability, cytotoxicity, lipid peroxidation, hydroxyl radicals, hydrogen peroxide, antioxidant enzyme levels and gene expressions, apoptotic, and oxidative stress-related gene expressions were measured in embryonic fibroblast cells. The results showed that GA induced cytotoxicity and diminished the expression levels of apoptotic genes. Furthermore, GA increased the levels of oxidative stress markers and significantly changed the oxidative stress-related gene expression. It has been determined that antioxidant molecules are considerably suppressed in GA-induced toxicity at both gene and enzyme levels. In addition to these results, when VitC, which is known to have strong antioxidant properties in eliminating the toxic effects of GA, is taken as reference, it has been proven that Hes has stronger antioxidant properties compared to VitC. Finally, GA-induced apoptosis in embryonic fibroblast cells is associated with nuclear factor erythroid 2-related factor 2 (Nrf2)-dependent oxidative stress and Hes has antioxidant properties with strong effects.

An assessment of the ameliorative role of hesperidin in Drosophila melanogaster model of cadmium chloride-induced toxicity.

Cadmium chloride (CdCl2) is an important heavy metal widely regarded as an environmental contaminant. Hesperidin, a flavanone glycoside found in citrus fruits, has an established properties against free radicals, apoptosis, and inflammation. The present study investigated the protective actions of hesperidin on CdCl2-induced oxidative damage and inflammation in Drosophila melanogaster. For 7 consecutive days via their diet regimen, the flies were exposed to CdCl2 alone (0.05 mM) or in combination with hesperidin (50 and 100 µM). Exposure to CdCl2 significantly (p < 0.05) increased mortality rate of flies, whereas the survived flies demonstrated significant oxidative toxicity from decreased activities of catalase and Glutathione S-transferase (GST) and Total Thiol (T-SH) and Non-Protein Thiols (NPSH) levels as well as accumulation of Nitric Oxide (NO (nitrite/nitrate)), protein carbonyl and Hydrogen Peroxide (H2O2). However, hesperidin-supplemented diet improved Acetylcholinesterase (AChE) activity, mitochondrial metabolic rate (cell viability), locomotor activity, and amelioration of oxidative damage and lipid peroxidation induced by CdCl2. The hesperidin diet supplement boosted the antioxidant milieu and ameliorated the oxidative damage in the treated flies. Overall, the findings revealed that hesperidin improved antioxidative protective capacity in Drosophila melanogaster model of CdCl2-induced toxicity. This suggests hesperidin as a potential therapeutic agent against oxidative stress disorders due to exposure to CdCl2 and or related toxicants.

Structural Investigation of Hesperetin-7-O-Glucoside Inclusion Complex with ß-Cyclodextrin: A Spectroscopic Assessment.

Flavonoids are biologically active natural products of great interest for their potential applications in functional foods and pharmaceuticals. A hesperetin-7-O-glucoside inclusion complex with ß-cyclodextrin (HEPT7G/ßCD; SunActive® HCD) was formulated via the controlled enzymatic hydrolysis of hesperidin with naringinase enzyme. The conversion rate was nearly 98%, estimated using high-performance liquid chromatography analysis. The objective of this study was to investigate the stability, solubility, and spectroscopic features of the HEPT7G/ßCD inclusion complex using Fourier-transform infrared (FTIR), Raman, ultraviolet-visible absorption (UV-vis), 1H- and 13C- nuclear magnetic resonance (NMR), differential scanning calorimetry (DSC), liquid chromatography/mass spectroscopy (LC-MS), scanning electron microscopy (SEM), and powdered X-ray diffraction (PXRD) spectroscopic techniques including zeta potential, Job's plot, and phase solubility measurements. The effects of complexation on the profiles of supramolecular interactions in analytic features, especially the chemical shifts of ß-CD protons in the presence of the HEPT7G moiety, were evaluated. The stoichiometric ratio, stability, and solubility constants (binding affinity) describe the extent of complexation of a soluble complex in 1:1 stoichiometry that exhibits a greater affinity and fits better into the ß-CD inner cavity. The NMR spectroscopy results identified two different configurations of the HEPT7G moiety and revealed that the HEPT7G/ßCD inclusion complex has both -2S and -2R stereoisomers of hesperetin-7-O-glucoside possibly in the -2S/-2R epimeric ratio of 1/1.43 (i.e., -2S: 41.1% and -2R: 58.9%). The study indicated that encapsulation of the HEPT7G moiety in ß-CD is complete inclusion, wherein both ends of HEPT7G are included in the ß-CD inner hydrophobic cavity. The results showed that the water solubility and thermal stability of HEPT7G were apparently increased in the inclusion complex with ß-CD. This could potentially lead to increased bioavailability of HEPT7G and enhanced health benefits of this flavonoid.

Assessment of in vitro bioaccessibility of carotenoids and phenolic compounds in a model milk-mandarine beverage.

Mandarine juice is one of the richest sources of ß-cryptoxanthin and flavonoids, which have been positively associated with bone mineral density. Carotenoids are lipophilic isoprenoid compounds with a complex absorption process that can be affected by different factors. In this study, we have evaluated the effect of the food matrix on the in vitro bioaccessibility of carotenoids and phenolic compounds in a model milk-mandarine beverage (MMB). MMBs were formulated with mandarine juice and different dairy products to achieve three fat levels (0.2%, 1.7% and 3.2%) and three calcium levels (120, 310 and 500 mg Ca2+ per 100 ml). The bioaccessibility was evaluated using a harmonised in vitro digestion method. The results showed that the content of milk fat increased the bioaccessibility in vitro of phenolic compounds (p < 0.05), while a moderate fat level (1.7%) resulted in the highest bioaccessibility for bioactive carotenoids. On the other hand, calcium fortification at the highest level (500 mg Ca2+ per 100 mL) decreased the bioaccessibility of bioactive carotenoids from 76% to 43% (66% for the major ß-cryptoxanthin) compared to the lower calcium fortification level (120 mg Ca2+ per 100 mL). The bioaccessibility of hesperidin, the main flavanone in mandarine juice, was significantly (p < 0.05) reduced in the MMB with the highest calcium level. The bioaccessibility of carotenoids and phenolic compounds is affected by fat and calcium levels. When formulating functional beverages, the impact of the formulation should be carefully considered to optimize the bioaccessibility of the bioactive compounds.

Solid self-microemulsifying nutraceutical delivery system for hesperidin using quality by design: assessment of biopharmaceutical attributes and shelf-life.

AIM: The present study endeavours to develop a solid self-microemulsifying nutraceutical drug delivery system for hesperidin (HES) using quality by design (QbD) to improve its biopharmaceutical attributes. METHODS: A 32 full factorial design was employed to study the influence of factors on selected responses. Risk assessment was performed by portraying Ishikawa fishbone diagram and failure mode effect analysis (FMEA). The in vivo antidiabetic study was carried on induced diabetic rats. RESULTS: The optimised liquid SMEDDS-HES (OF) formulation showed emulsification time (Y 1) = 102.5 ± 2.52 s, globule size (Y 2) = 225.2 ± 3.40 nm, polydispersity index (Y 3) = 0.294 ± 0.62, and zeta potential (Y 4) = -25.4 ± 1.74 mV, respectively. The solid SMEDDS-HES (SOF-7) formulation was characterised by FTIR, PXRD, DSC, and SEM. The shelf life of SOF-7 was found to be 32.88 months. The heamatological and histopathological data of diabetic rats showed prominent antidiabetic activity. CONCLUSIONS: The optimised formulation showed improved dissolution, desired stability, and promising antidiabetic activity.

Biochemical and genotoxic biomarkers and cell cycle assessment in the zebrafish liver (ZF-L) cell line exposed to the novel metal-insecticide magnesium-hespiridin complex.

The flavonoid metal-insecticide magnesium-hesperidin complex (MgHP) has recently been considered as a novel insecticide to replace some persistent pesticides. However, it is important to evaluate its action on non-target species, mainly those living in an aquatic environment, as these ecosystems are the final receptors of most chemicals. Reactive oxygen species, antioxidant and oxidative stress biomarkers, genotoxicity as well as cell cycle was evaluated in the liver cell line from zebrafish (Danio rerio; ZF-L) exposed to 0, 0.1, 1, 10, 100 and 1000 ng mL-1 MgHP. MgHP affected cell stability by increasing reactive oxygen species (ROS) in both exposure times (24 and 96 h) at high concentrations. Catalase (CAT) activity decreased after 24 h exposure, and glutathione and metallothionein values increased, avoiding the lipid peroxidation. Genotoxicity increased as MgHP concentration increased, after 24 h exposure, exhibiting nuclear abnormalities; it was recovered after 96 h exposure, evidencing possible stimulation of DNA repair mechanisms. The alteration in the cell cycle (increasing in the Sub-G1 phase and decreasing in the S-phase) was associated with chromosomal instability. In conclusion, the responses of ROS and the antioxidant defense system depended on MgHP concentration and time exposure, while DNA exhibited some instability after 24 h exposure, which was recovered after 96 h.

Radiation shielding parameters of some antioxidants using Monte Carlo method.

In this paper, radiation shielding parameters such as mass attenuation coefficients and half value layer (HVL) of some antioxidants are investigated using MCNPX (version 2.4.0). The validation of the generated MCNPX simulation geometry for antioxidant structures is provided by comparing the results with standard WinXcom data for radiation mass attenuation coefficients of antioxidants. Very good agreement between WINXCOM and MCNPX was obtained. The results from the validated geometry were used to calculate the shielding parameters of different antioxidants. The radiation attenuation properties of each antioxidant were compared with each other. The results showed that, on average, the highest and the lowest radiation mass attenuation coefficients were observed on hesperidin and delphinidin chloride, respectively. It can be concluded that Monte Carlo simulation is a strong tool and an alternate method where experimental investigations are not possible and a standard simulation setup can be used in further studies for different biological structures. It can also be concluded that the obtained results from this study are very useful for radiology and radiotherapy applications where antioxidants are frequently used.

Simple low-cost miniaturization approach for pharmaceutical nanocrystals production.

Systematic screening for optimal formulation composition and production parameters for nanosuspensions consumes a lot of time and also drug material when performed at lab scale. Therefore, a cost-effective miniaturized scale top down approach for nanocrystals production by wet bead milling was developed. The final set-up consisted of 3 magnetic stirring bars placed vertically one over the other in a 2 mL glass vial and agitated by a common magnetic stirring plate. All of the tested actives (cyclosporin A, resveratrol, hesperitin, ascorbyl palmitate, apigenin and hesperidin) could be converted to nanosuspensions. For 4 of them, the particles sizes achieved were smaller than previously reported on the literature (around 90 nm for cyclosporin A; 50 nm for hesperitin; 160 nm for ascorbyl palmitate and 80 nm for apigenin). The "transferability" of the data collect by the miniaturized method was evaluated comparing the production at larger scale using both wet bead milling and high pressure homogenization. Transferable information obtained from the miniaturized scale is minimum achievable size, improvements in size reduction by reduction of beads size, diminution kinetics and potentially occurring instabilities during processing. The small scale batches also allow identification of optimal stabilizer types and concentrations. The batch size is 0.5 mL, requiring approximately 50 mg or 5 mg of drug (5% and 1% suspension, respectively). Thus, a simple, accessible, low-cost miniaturized scale method for the production of pharmaceutical nanocrystals was established.

Diosmina em combinação com a hesperidina para o tratamento da doença venosa crônica / Diosmina in combination with hesperidin for the treatment of chronic venous disease

CONTEXTO: A doença venosa crônica (DVC) é proveniente de uma série de distúrbios morfológicos e anatômicos que ocorrem nas veias, interferindo na pressão venosa e no retorno do sangue periférico até o coração e se manifestam a partir do surgimento de veias dilatadase tortuosas (vasos e varizes) até a formação de coágulos no interior das veias (trombose venosa). Os sinais e os sintomas da DVC estão relacionados com o local de acometimento, o tempo e a gravidade da doença. Quando a doença se complica e os sinais e sintomas avançam, ela é denominada insuficiência venosa crônica (IVC). O seu tratamento consiste em medidas conservadoras, como o uso de meias elásticasno caso da DVC nas pernas, medicamentos, escleroterapia e cirurgias. TECNOLOGIA: Diosmina em combinação com a hesperidina. PERGUNTA: Existe evidência para o uso de diosmina em combinação com hesperidina para o tratamento da doença venosa crônica? EVIDÊNCIAS: Foram encontrados um ensaio clínico randomizado, que avaliou a combinação entre diosmina e hesperidina, e uma revisão sistemática, que avaliou a combinação entre diosmina e hidrosmina. Os resultados do ensaio clínico randomizado demonstraram que o uso de diosmina associado com hesperidina não apresentou evidências satisfatórias para proporcionar alívio dos sintomas e melhora da qualidade de vida em pacientes com doença venosa crônica. Os resultados da revisão sistemática sugerem eficácia limitada da diosmina associada à hidrosmina na redução de edema sem influenciar na cicatrização de úlceras. CONCLUSÕES: As evidências disponíveis não fornecem embasamento de forma conclusiva para o uso de diosmina associada com hesperidina para o tratamento da doença venosa crônica. Novos estudos com alta qualidade metodológica devem ser realizados a fim de suportar o uso desta associação.

Avaliação preliminar da associação entre anfotericina e hesperina com oxidantes e células huvec / Preliminary assessment of the association between amphotericin and hesperetin with oxididants and huvec cells

A anfotericina B (AmB) é fármaco o “padrão ouro” para o tratamento de infecções fúngicas invasivas desde 1960, Entretanto, a anfotericina B apresenta elevada toxicidade, a qual manifesta-se mais frequentemente nos rins e no fígado, Sabe-se, desde 1985, que a auto-oxidação da AmB origina diferentes formas de espécies reativas oxidativas e estas, por serem tóxicas para a célula, seriam responsáveis, em parte, pela toxicidade, Diferentes estudos indicam que a hesperidina contribui por meio do decréscimo do estresse oxidativo, para a proteção renal e contra a injúria gerada pela isquemia, Tal fato e o envolvimento da AmB na geração de radicais livres tornam interessante a avaliação preliminar do efeito da hesperidina e AmB (isoladamente ou associadas) frente a espécies reativas do oxigênio e radicais livres, bem como o estudo das mesmas em modelos de citoxicidade, Frente ao ABTS•+, a AmB apresentou IC50 igual a 0,0124mg/mL, mas quando associada à hesperidina este valor caiu para 0,0003mg/mL, Frente ao HOCl, a Amb apresentou IC50 igual a 0,0056, mas quando associada à hesperidina este valor caiu para 0,0023mg/mL, No ensaio com DPPH• e radical ânion superóxido as amostras não foram efetivas, No ensaio com células endoteliais em cultivo (HUVEC), as associações reduziram a viabilidade celular, Estes resultados preliminares evidenciam a interação dos compostos com espécies reativas bem como indicam possibilidade de exacerbação do dano pela AmB na presença dos antioxidantes em um modelo in vitro...

Amphotericin B (AmB) is drug “gold standard” for the treatment of invasive fungal infections since 1960. However, amphotericin B has high toxicity, which manifests itself most often in the kidneys and in the liver. It is known, since 1985, that self-oxidation of AmB gives different forms of reactive oxidative species and these, being toxic to the cell, would be responsible, in part, by its toxicity. Different studies indicate that hesperidin contributes, through the reduction of oxidative stress, to protect against renal injury generated by ischemia. This fact and the involvement of AmB in the generation of free radicals make it interesting the preliminary evaluation of the effect of hesperidin and AmB (alone or associated) against reactive oxygen species and free radicals, as well as the study on models of cytotoxicity. Front ABTS•+, AmB presented IC50 equal to 0.0124 mg/mL, but when it was associated to hesperidin this value has decreased to 0.0003 mg/mL. Front HOCl, Amb presented IC50 equal to 0.0056, but when it was associated to hesperidin this value has decreased to 0.0023 mg/mL. In the trials with DPPH• and the superoxide anion radical samples were not effective. In the assay with endotelial cell culture (HUVEC cells), the association has decreased cell viability. These preliminary results demonstrate the interaction of the compounds with reactive species as well as indicate the possibility of damage exacerbation by AmB in the presence of antioxidants in an in vitro model...

[Ultrasound assessment of alterations in venous haemodynamics in patients with post-thrombotic disease permanently taking phlebotonics].

OBJECTIVE: The study was aimed at using ultrasound duplex scanning for determining the sequence and terms of formation of venous haemodynamics impairments in the affected lower extremity in patients after endured acute thrombosis of deep veins and assessing the effect of phlebotonic drugs on the course of these processes. MATERIAL AND METHODS: We examined and treated a total of 66 patients presenting with newly onset acute thrombosis of deep veins of lower limbs without concomitant varicose disease. Group I patients (n = 22) received the standard course of angiotropic and metabolic infusion therapy, direct and indirect anticoagulants, as well as used elastic compression. Group II patients (n = 22) in addition to the similar course of treatment received a phlebotonic drug (Venarus) according to the standard regimen: 1,000 mg daily for two months every half year. Group III patients (n = 22) additionally to the same standard treatment regimen were also given VenarusR at a dose of 1,000 mg daily but taken uninterruptedly and constantly during the whole period of follow up. All patients were subjected to ultrasound duplex scanning of deep veins of lower limbs initially at admission, then 3 weeks, 3, 6, 12 and 18 months after making the diagnosis of acute thrombosis. RESULTS: Group II and III patients additionally taking the phlebotonic were found to have acceleration of processes of recanalization averagely by 15-20% as compared with Group I patients. Group III patients taking the phlebotonic agent permanently demonstrated deceleration of the processes of formation of horizontal and vertical veno-venous refluxes on the background of more adequate recanalization by the end of the follow-up period. CONCLUSION: Permanent taking of phlebotonics increases the rate and scope of recanalization of the thrombosed deep veins of lower limbs, as well as dramatically decreases the development of the horizontal and vertical reflux, decreasing clinical manifestations of chronic venous insufficiency.

Simplified miniaturized ultrasound-assisted matrix solid phase dispersion extraction and high performance liquid chromatographic determination of seven flavonoids in citrus fruit juice and human fluid samples: hesperetin and naringenin as biomarkers.

In the present study, for the first time, a simplified miniaturized ultrasound-assisted matrix solid-phase dispersion (SM-USA-MSPD) method with a different application for liquid matrices was developed to extract different flavonoids (hesperidin, diosmin, eriocitrin, narirutin, naringin, hesperetin and naringenin) from citrus fruit juice and human fluid samples prior to their determination using high performance liquid chromatography (HPLC). Different effective parameters were studied and under the optimum conditions (including sample volume: 150µL; solid phase: silica-based C18, 200mg; eluting solvent: methanol, 500µL; pH: 4; and sonication: 6min; at room temperature), limits of detection and limits of quantification were ranged from 23.3 to 46.8ngmL(-1) and 74.8 to 141.5ngmL(-1), respectively. Once optimized, analytical performance of the method was studied in terms of linearity (0.074-198.5µgmL(-1), r(2)>0.991), accuracy (recovery=84.6-101.5%), and precision (repeatability: intra-day precision<5.9%, and inter-day precision<7.2%). At the end, SM-USA-MSPD method was successfully applied to estimate the levels of hesperetin and naringenin in plasma and urinary excretion -after ingestion of orange, grapefruit and lime juices- and the obtained results confirmed that these compounds could be used as good biomarkers of citrus fruit juice intake.

Assessment of phytochemical content in human milk during different stages of lactation.

OBJECTIVE: The present study reports the presence of several carotenoids and flavonoids in human milk samples. METHODS: Samples were collected from 17 women who delivered healthy term babies (≥ 37 wk of gestation) at 1-, 4-, and 13-wk postpartum intervals. RESULTS: Epicatechin (63.7-828.5 nmol/L), epicatechin gallate (55.7-645.6 nmol/L), epigallocatechin gallate (215.1-2364.7 nmol/L), naringenin (64.1-722.0 nmol/L), kaempferol (7.8-71.4 nmol/L), hesperetin (74.8-1603.1 nmol/L), and quercetin (32.5-108.6 nmol/L) were present in human milk samples with high inter-/intraindividual variability. With the exception of kaempferol, the mean flavonoid content in human milk was not statistically different among lactation stages. In contrast, carotenoids α-carotene (59.0-23.2 nmol/L), ß-carotene (164.3-88.0 nmol/L), α-cryptoxanthin (30.6-13.5 nmol/L), ß-cryptoxanthin (57.4-24.8 nmol/L), zeaxanthin (46.3-21.4 nmol/L), lutein (121.2-56.4 nmol/L), and lycopene (119.9-49.5 nmol/L) significantly decreased from weeks 1 to 13 of lactation. CONCLUSION: The observed differences in the relative concentrations of the two phytochemical classes in human milk may be a result of several factors, including dietary exposure, stability in the milk matrix, efficiency of absorption/metabolism, and transfer from plasma to human milk. These data support the notion that flavonoids, as with carotenoids, are dietary phytochemicals present in human milk and potentially available to breast-fed infants.

Assessment of the antineoplastic potential of chalcones in animal models.

One part of chemical space that is endowed with interesting biological properties is the area of the chalcones. With this review, we provide a comprehensive overview of the numerous in vivo animal studies on the antineoplastic potential of both natural and synthetic members of this flavonoid subclass (covering: up to mid-2011). The thus far identified modes of action of these compounds are also discussed. We hope that this overview may stimulate deeper investigations into the biochemical mechanisms by which chalcones exert their antineoplastic action. As a result, in the foreseeable future, chalcones may prove suitable lead molecules or early drug candidates for the prevention or treatment of various neoplastic diseases.

Genotoxicity assessment of Pyungwi-san (PWS), a traditional herbal prescription.

ETHNOPHARMACOLOGICAL RELEVANCE: Pyungwi-san (PWS, Heii-san in Japanese) is a mixture of six herbs and is traditionally used in Northeast Asia (especially Korea and Japan) for the treatment of gastrointestinal disorder, such as dyspepsia and inappetance induced by gastric dilatation and gastrointestinal catarrh. AIM OF THE STUDY: Although PWS is a widely used herbal prescription in Korea and Japan, little information is available in the literature on the safety and toxicity of PWS. As part of a safety evaluation of PWS, the present study evaluated the potential genotoxicity of PWS using a standard battery of test. MATERIALS AND METHODS: We prepared PWS using a water extraction method and simultaneously extracted three compounds from PWS using high performance liquid chromatography. The PWS extract that was obtained was assayed for genotoxicity using the standard three tests recommended by the Korea Food and Drug Administration. These tests included the bacterial reverse mutation test (Ames test), the chromosomal aberration test using China hamster lung cells, and the micronucleus test using ICR mice. RESULTS: The Ames test showed that the PWS extract did not induce an increase in the number of revertant colonies compared with vehicle control at any dose in all of tester strains. In the micronucleus test, no significant increase was observed in micronucleated polychromatic erythrocytes (MNPCEs) at any dose of PWS extract compared with vehicle control. Conversely, chromosomal aberration test showed that the PWS extract at a dosage of 4500 µg/mL induced an increase in the number of chromosomal aberrations in the 6 h group with metabolic activation compared with the vehicle control. CONCLUSION: PWS extract exhibits genotoxicity, based on the results of the chromosomal aberration test. Thus, further detailed experiments will be needed to identify the ingredient responsible for inducing this genotoxicity and to determine its mechanism.

Assessment of quality of life in Mexican patients suffering from chronic venous disorder - impact of oral Ruscus aculeatus-hesperidin-methyl-chalcone-ascorbic acid treatment - 'QUALITY Study'.

OBJECTIVES: The present study assessed the effect of Ruscus aculeatus-hesperidin-methyl-chalcone-ascorbic acid (HMC-AA) on the quality of life (QoL) of patients suffering from chronic venous disorders (CVDs). METHODS: An observational, multicentre and prospective study was performed with 917 Mexican patients suffering from CVD. Patients were treated with R. aculeatus-HMC-AA. After 12 weeks of treatment, the physicians then assessed the patients' symptoms and QoL using Short Form (SF-12) and Chronic Venous Insufficiency (CIVIQ) auto-questionnaires. RESULTS: Patients were mainly women (86.7%), overweight or obese (72.7%) or C2 (39.3%)-C3 (27.6%). All symptoms and ankle circumferences significantly improved over time, with increasing clinical, aetiological, anatomical and pathophysiological (CEAP) classes and body mass index (BMI) (P < 0.001). Concerning QoL, all dimensions of the SF-12 score significantly improved over time (P < 0.001). Moreover, the CIVIQ scores significantly improved (P < 0.001) with increasing BMI (P < 0.002) and CEAP classes (P < 0.05). CONCLUSION: R. aculeatus-HMC-AA significantly improved the symptoms and QoL of CVD patients.