The chemical characterisation of nettle (Urtica dioica L.) seed oil.

The past decade has seen increasing interest in plant oils in the food, pharmaceutical, and cosmetic industries due to their diverse functional and nutritional properties, as well as the high levels of consumer acceptance. As a result, there has been a spike in research interest in finding new sources of oils with good potential for industrial use. In this paper, the chemical characterisation (saponification, iodine, acid and ester numbers, as well as fatty acid composition) of nettle seed oil was given. Oil was analysed for its thermooxidative stability by differential scanning calorimetry. The kinetic parameters of oxidation (Ea, A, and k) were also determined. The results showed that nettle seed oil belongs to the group of edible oils of the linoleic type and it is nutritionally highly valuable, it has pronounced thermooxidative stability and could be an alternative source of raw material for the food, cosmetics, and other non-food industries.

Characterization and application of biochar-immobilized crude horseradish peroxidase for removal of phenol from water.

Biochar (BC) has attracted much attention as an environmentally friendly material for application in wastewater treatment. In this study, a suitability of wood-derived BC as a support for covalent immobilization of horseradish peroxidase (HRP) across glutaraldehide as crosslinker, known for the capability to remove phenol from water, was investigated. The efficiency of the immobilized HRP in removal of phenol (2 mM) from water at different reaction conditions (varying dosages of polyethylene glycol (PEG300) 0-750 mg/L; H2O2 1.5-3.5 mM, as well as reaction time 5-120 min) and the general toxicity of bio-treated water (Allium cepa test) were measured. All analyzes were performed for free enzyme as well. The immobilized enzyme showed the highest activity at temperature 30 °C and pH 7.0. The greatest efficiency of immobilized enzyme in phenol removing (90 %) was obtained by applying 2.5 mM H2O2 and 1.5 mg/L of PEG300 at pH 7.0 after 2 h of reaction period. After 4 washings, immobilized HRP retained more than 79 % activity with phenol removal of 64 %. Utilizing immobilized enzyme significantly reduces the toxicity of the tested water (80 %), which further suggested that it might be considered as an environmentally acceptable process for wastewater treatment. Possible degradation products remained in treated water were analyzed in water samples by liquid and gas chromatography with mass spectrometry, including also analysis of volatiles by solid phase microextraction technique; different phenol-base compounds were detected.

'All-natural' anti-wrinkle emulsion serum with Acmella oleracea extract: A design of experiments (DoE) formulation approach, rheology and in vivo skin performance/efficacy evaluation.

OBJECTIVE: The growing consumers' preferences and concerns regarding healthy ageing, youthful skin appearance, environmental protection and sustainability have triggered an ever-increasing trend towards natural, eco-friendly and ethically sourced anti-ageing products. Accordingly, this paper describes design and evaluation of novel, safe, effective and high-quality emulsion serums, completely based on ingredients of natural origin, intended for improving facial fine lines and wrinkles. METHODS: Model formulations, stabilized by an innovative glycolipid mixed emulsifier (lauryl glucoside/myristyl glucoside/polyglyceryl-6 laurate) and containing Acmella oleracea extract as a model anti-ageing active, were prepared by cold process and fully assessed regarding their rheological behaviour (continuous rotational and oscillatory tests) and physical stability (dynamic-mechanical thermoanalysis - DMTA test). To study and optimize the simultaneous influence of varied formulation factors (emollients and emulsifier concentrations) on critical rheological attributes of the developed serums, a central composite design within 'design of experiments' approach was employed. The general skin performance - preliminary safety and anti-wrinkle efficacy of selected model serum, was evaluated in human volunteers, by employing several objective, non-invasive bioengineering techniques. RESULTS: Rheological characterization revealed favourable shear-thinning flow behaviour with yield point, and dominating elastic character (storage modulus G' > loss modulus G") in both amplitude and frequency sweeps, which together with relatively small structural change obtained in DMTA test indicated overall satisfying rheological and stability properties of formulated serums. From the established design space, and taking into account formulation cost and carbon footprint, promising model serum (desired/optimal apparent viscosity, yield point and loss factor, rather small and constant structural change), containing 15% of emollients and 1% of emulsifier, was chosen for in vivo evaluations. Screening of skin irritation effects revealed the absence of potential irritancy of investigated serum, suggesting overall satisfying skin tolerability/preliminary safety. Silicone skin replica image analysis demonstrated noticeable reduction/improvement in all measured skin wrinkle parameters after only 2 weeks of test serum application in periorbital and perioral areas, indicating its rapid and beneficial effects on the facial expression lines and wrinkles. CONCLUSION: Altogether, the results corroborate the promising potential of the developed Acmella oleracea extract-loaded emulsion serum as safe, effective and non-invasive natural anti-wrinkle product.

OBJECTIF: Les préférences et les préoccupations croissantes des consommateurs concernant le vieillissement sain, l'apparence jeune de la peau, la protection de l'environnement et la durabilité ont déclenché une tendance toujours croissante vers des produits anti-âge naturels, respectueux de l'environnement et éthiques. En conséquence, ce document décrit le plan et l'évaluation de nouveaux sérums d'émulsion sûrs, efficaces et de haute qualité, entièrement basés sur des ingrédients d'origine naturelle, destinés à améliorer les ridules et rides du visage. MÉTHODES: Des formulations modèles stabilisées par un émulsifiant mixte glycolipide innovant (lauryl glucoside/myristyl glucoside/polyglycéryl-6 laurate) et contenant de l'extrait d'Acmella oleracea comme anti-vieillissement actif de modèle, ont été préparées par un procédé à froid et ont été pleinement évaluées en ce qui concerne leur comportement rhéologique (tests de rotation continue et examens oscillatoires) et stabilité physique (analyse thermomécanique dynamique - DMTA). Pour étudier et optimiser l'influence simultanée de facteurs de formulation variés (concentrations d'émollients et d'émulsifiants) sur les attributs rhéologiques critiques des sérums développés, une conception composite centrale dans le cadre d'une approche « conception d'expériences ¼ a été employée. Les performances cutanées générales - sécurité préliminaire et efficacité antirides du sérum du modèle sélectionné ont été évaluées chez des sujets humains volontaires, en utilisant plusieurs techniques de bio-ingénierie objectives et non invasives. RÉSULTATS: La caractérisation rhéologique a révélé un comportement favorable du débit de cisaillement avec une limite de rendement et une domination du caractère élastique (modulus de stockage G' > module de perte G) dans les balayages d'amplitude et de fréquence, qui, avec un changement structurel relativement faible obtenu dans l'analyse DMTA, a indiqué des propriétés rhéologiques et de stabilité satisfaisante globales des sérums. A partir de l'espace de conception établi, et en tenant compte du coût de composition et de l'empreinte carbone, un sérum modèle prometteur (viscosité apparente souhaitée/optimale, seuil de rendement et facteur de perte, changement structurel assez faible et constant), contenant 15 % d'émollients et 1 % d'émulsifiant, a été choisi pour les évaluations in vivo. Le dépistage des effets d'irritation cutanée a révélé l'absence d'irritation potentielle du sérum expérimental, suggérant une tolérance cutanée/une sécurité préliminaire globalement satisfaisante. L'analyse de l'image de la réplique cutanée en silicone a démontré une réduction/amélioration notable de tous les paramètres de rides cutanées mesurés après seulement deux semaines d'application du sérum test dans les zones périorbitaires et péribuccales, indiquant ses effets rapides et bénéfiques sur les lignes d'expression et les rides du visage. CONCLUSION: Au total, les résultats corroborent le potentiel prometteur du sérum d'émulsion à base d'extrait d'Acmella oleracea développé comme un produit anti-rides naturel sûr, efficace et non invasif.

Identification and photostability of N-alkylamides from Acmella oleracea extract.

The identification of N-alkylamides from commercial Acmella oleracea extract, their UV-B photostability in different solvents, and identification of degradation products were the main goals of this study. By UHPLC-DAD-ESI-MS/MS method the presence of nine N-alkylamides was identified. Investigation of UV-B irradiation effect on identified N-alkylamides from Acmella oleracea extract was monitored in various the most commonly used solvents (methanol, ethanol, saline solution, and water) during 120 min. The results obtained indicated that spilanthol and homospilanthol were the most stable N-alkylamides presented in Acmella oleracea extract, while the photostability of identified N-alkylamides in whole in tested extract solutions decreased as follows: methanol>ethanol>saline solution>water. As the main degradation products in all investigated solutions 6,9-dihydroxy-deca-2,7-dienoic acid isobutyl-amide and 8,9-dihydroxy-deca-2,6-dienoic acid isobutyl-amide were identified.

Rutin as Deoxyribonuclease I Inhibitor.

DNase I inhibitory potential of water extract of nine Hypericum species (H. umbellatum, H. barbatum, H. rumeliacum, H. rochelii, H. perforatum, H. tetrapterum, H. olympicum, H. hirsutum, H. linarioides) and the most important Hypericum secondary metabolites (hypericin, hyperforin, quercetin, and rutin) was investigated. All examined Hypericum extracts inhibited DNase I with IC50 below 800 µg/ml, whereby H. perforatum was the most potent (IC50 =391.26±68.40 µg/ml). Among the investigated Hypericum secondary metabolites, rutin inhibited bovine pancreatic DNase I in a non-competitive manner with IC50 value of 108.90±9.73 µm. DNase I inhibitory ability of rutin was further confirmed on DNase I in rat liver homogenate (IC50 =137.17±16.65 µm). Due to the involvement of DNase I in apoptotic processes the results of this study indicate the importance of frequent rutin and H. perforatum consumption in daily human nutrition. Rutin is a dietary component that can contribute to male infertility prevention by showing dual mechanism of sperm DNA protection, DNase I inhibition and antioxidant activity.

Autofermentation of Chamomile Ligulate Flowers Promote Antitumor Effects in vitro.

In the frame of this paper, the enzyme-assisted hydrolysis coupled with ultrasound and Soxhlet extraction was applied in order to get extracts of chamomile ligulate flowers (CLF). Obtained extracts were characterized in terms to their apigenin and apigenin glucoside composition, as well as antiproliferative potential against tumour cells. Antioxidant activity was determined by two different assays based on different mechanisms showing that autofermented extracts have higher reduction potential. Autofermented extracts prepared by ultrasound and Soxhlet extraction had a stronger impact on the treated carcinoma (HeLa and NCI-H358) and leukemia (K562) cells' growth reduction in comparison to the native extracts, 30-35% greater inhibition at the lowest concentration (0.01 mg/mL), in two observed time points (48 and 72 h). Leukemia cells are more sensitive to all tested extracts. The autofermented CLF extracts with highest antiproliferative efficacy induced morphological changes and apoptosis in the HeLa cells. Obtained results clearly showed that the combination of enzymatic hydrolysis with cavitation phenomenon results in extracts with higher apigenin content and increased biological potential.

Sulfanilamide in solution and liposome vesicles; in vitro release and UV-stability studies.

The main goal of this study was to develop a liposome formulation with sulfanilamide and to investigate the liposomes impact on its release and stability to the UV-A/UV-B and UV-C irradiation. Liposome dispersions with incorporated sulfanilamide were prepared by thin-film hydration method and liposomes role to the sulfanilamide release was investigated by using a dialysis method. Comparatively, sulfanilamide in phosphate buffer solution was subject to release study as well to the UV irradiation providing for the possibilities of kinetics analysis. In vitro drug release study demonstrated that 20% of sulfanilamide was released from liposomes within 1 h that is approximately twice as slower as in the case of dissolved sulfanilamide in phosphate buffer solution. The kinetic release process can be described by Korsmeyer-Peppas model and according to the value of diffusion release exponent it can be concluded that drug release mechanism is based on the phenomenon of diffusion. The sulfanilamide degradation in phosphate buffer solution and liposomes is related to the formation of UV-induced degradation products that are identified by UHPLC/MS analysis as: sulfanilic acid, aniline and benzidine. The UV-induced sulfanilamide degradation in the phosphate buffer solution and liposome vesicles fits the first- order kinetic model. The degradation rate constants are dependent on the involved UV photons energy input as well as sulfanilamide microenvironment. Liposome microenvironment provides better irradiation sulfanilamide stability. The obtained results suggest that liposomes might be promising carriers for delayed sulfanilamide delivery and may serve as a basis for further research.

Parenteral nanoemulsions as promising carriers for brain delivery of risperidone: Design, characterization and in vivo pharmacokinetic evaluation.

This paper describes design and evaluation of parenteral lecithin-based nanoemulsions intended for brain delivery of risperidone, a poorly water-soluble psychopharmacological drug. The nanoemulsions were prepared through cold/hot high pressure homogenization and characterized regarding droplet size, polydispersity, surface charge, morphology, drug-vehicle interactions, and physical stability. To estimate the simultaneous influence of nanoemulsion formulation and preparation parameters--co-emulsifier type, aqueous phase type, homogenization temperature--on the critical quality attributes of developed nanoemulsions, a general factorial experimental design was applied. From the established design space and stability data, promising risperidone-loaded nanoemulsions (mean size about 160 nm, size distribution <0.15, zeta potential around -50 mV), containing sodium oleate in the aqueous phase and polysorbate 80, poloxamer 188 or Solutol(®) HS15 as co-emulsifier, were produced by hot homogenization and their ability to improve risperidone delivery to the brain was assessed in rats. Pharmacokinetic study demonstrated erratic brain profiles of risperidone following intraperitoneal administration in selected nanoemulsions, most probably due to their different droplet surface properties (different composition of the stabilizing layer). Namely, polysorbate 80-costabilized nanoemulsion showed increased (1.4-7.4-fold higher) risperidone brain availability compared to other nanoemulsions and drug solution, suggesting this nanoemulsion as a promising carrier worth exploring further for brain targeting.

Comparative analysis of antioxidant, antimicrobiological and cytotoxic activities of native and fermented chamomile ligulate flower extracts.

MAIN CONCLUSION: The work investigated differences in apigenin content, as well as in other compounds, and examined the chemical profiles, antioxidant, antimicrobial and cytotoxic effects of extracts obtained from native and fermented chamomile ligulate flowers. Chamomile (Chamomilla recutita L.) has a long history of being used as a medicinal plant due to many health benefits, including antiinflammatory, anticancer, antispasmodic, radical-scavenging effects and others. Apigenin is recognized as one of the most bioactive phenolic compounds in chamomile. In comparison to its bound forms, which include mostly apigenin-7-O-ß-glucoside and various acylated forms, the aglycone is attributed with much higher bioactivity. Due to this fact, in this work ligulate florets of chamomile anthodium were subjected to a fermentation process using native chamomile enzymes to hydrolyze bound forms of apigenin to free aglycone. The contents of apigenin and apigenin-7-O-ß-glucoside were determined in both fermented and nonfermented samples by UHPLC-MS-MS analysis to define the efficiency of conversion. After defining their chemical profiles, the extracts of fermented and nonfermented chamomile samples were also compared with respect to their antioxidant, antimicrobial and cytotoxic effects. The antioxidant effects of the obtained extracts were defined by electron spin resonance analysis for hydroxyl and superoxide radicals. The antimicrobial activity was defined for eight microbial strains, whereas cytotoxic activity was evaluated using two human cell lines (human cervix carcinoma and human rhabdomyosarcoma) and murine fibroblasts.

Enzymatic oxidation of rutin by horseradish peroxidase: kinetic mechanism and identification of a dimeric product by LC-Orbitrap mass spectrometry.

Flavonoid oxidation is important issue in food processing and quality. The kinetic mechanism of enzymatic oxidation of rutin by horseradish peroxidase (HRP) was studied. Rutin oxidation reaction was followed by recording of spectral changes over the time at 360 nm. The studied oxidation is mostly enzymatic and less part non-enzymatic. The reaction with HRP has a higher rate compared with the reaction without of HRP, whereby is part of non-enzymatic reaction about 10% of the total reaction. Kinetic parameters were determined from graphics of linear Michaelis-Menten equation, and it was found that investigated reactions of rutin oxidation by HRP take place in a ping-pong kinetic mechanism. High resolution HPLC-MS analysis of the mixture of oxidized products of rutin revealed the presence of rutin dimer. Because of widely distribution of rutin as well as presence of peroxidases and hydrogen peroxide in fresh foods identification of this enzymatic modification product can be beneficial for foods quality and safety.

Kinetics of enzymatic modification of quercetin with cysteine by horseradish peroxidase.

The kinetic mechanism of enzymatic modification of flavonol quercetin with L-cysteine by horseradish peroxidase (HRP) was studied. Reaction of modification of quercetin was followed by recording spectral changes over time at 380 nm. All reactions were performed in 100 mM phosphate buffer pH, 6.0 at 20 degrees C. Kinetic parameters were determined from graphics of linear Michaelis-Menten equation. The values obtained at specified intervals were: V(max) = 0.17 divided by 0.91 deltaA380/min, K(m) = 0.023 divided by 0.5 mM, k(cat) = 0.21 divided by 1.14 deltaA380/min x nM(-1) and V(max)/K(m) = 0.83 divided by 26.55 deltaA380/min x mM(-1). It was found that all investigated reactions of the modification of quercetin with L-cysteine by HRP followed an ordered mechanism. We propose that HRP initially reacts with H2O2 than with quercetin and finally with L-cysteine, leading to the introduction of L-cysteine in the structure of quercetin.

Hemodynamics of arterial and venous circulation in the intrauterine fetal evaluation.

INTRODUCTION: The purpose of each antenatal control focuses on the detection and prevention of hypoxic-ischemic injury and fetal death (1)) using modern biophysical tests and Doppler parameters. GOAL: This study examines the correlation of changes in arterial compared to venous hemodynamics of the fetus and is determined by the most sensitive and most specific Doppler parameter in the assessment of intrauterine fetal status. MATERIAL AND METHODS: The study was conducted as prospective and included 119 pregnant women. All subjects underwent NST (nonstres test), and Doppler measurements of blood flow in the umbilical artery (Aum), fetal aorta (Ao) and the central cerebral artery (MCA). In case of borderline and pathological arterial flow was measured through the ductus venozus (DV) and umbilical vein (VU). At birth to the child was determined umbilical artery blood pH and Apgar score (AS) in the first minute. Based on the clinical condition of the newborn and outcome was calculated perinatal morbidity and mortality. RESULTS: In all cases with a pathological arterial flow, which is verified during measurement also the pathological venous flow and confirmed fetal acidemia at birth and low Apgar scores? In this group, the two neonates died in the first week. CONCLUSION: There is a justification for the analysis of venous flow in the event borderline and pathologic findings in fetal arterial system. It has been proven that the cerebroumbilical (C/U) index is most effective parameter in predicting changes in the venous system and this the most sensitive Doppler parameter in predicting fetal acidosis and the most specific Doppler flow through the central cerebral artery.

Clinical and prognostic significance of apoptotic profile in patients with newly diagnosed nodal diffuse large B-cell lymphoma (DLBCL).

BACKGROUND: Apoptosis-related proteins might play an important role in the pathogenesis of lymphoma and sensibility to chemotherapy (CH) in patients with non-Hodgkin's lymphoma. We have analyzed the relationship between expression of two proapoptotic (CD95, caspase-3) and four antiapoptotic proteins (c-FLIP, bcl-2, survivin, and XIAP) and clinical outcome of patients with nodal diffuse large B-cell lymphoma (DLBCL). METHODS: We have analyzed lymph node biopsy specimens obtained from 78 patients with newly diagnosed nodal DLBCL. The expression of apoptotic parameters was analyzed using the standard immunohistochemical method (antibodies against caspase-3, CD95, c-FLIP, XIAP, survivin, and bcl-2) on formalin-fixed and routinely processed paraffin-embedded lymph node specimens. The expression of immunohistochemical parameters has been evaluated semiquantitatively as a percentage of tumor cells. RESULTS: Immunoexpression of caspase-3, CD95, c-FLIP, survivin, XIAP, and bcl-2 has been found in 48 (61.5%), 39 (50%), 45 (57.7%), 41 (52.6%), 43 (55.12%), and 39 (50.0%) patients, respectively. The therapy response was achieved in 53 (67.9%) patients. Besides numerous clinical parameters, survivin and XIAP positivity along with CD95 negativity were found to be unfavorable factors for therapy response and shorter survival in univariate analysis. According to this finding, an 'apoptotic score' that includes unfavorable apoptotic parameters has been defined. In multivariate analysis, only International Prognostic Index (IPI) and apoptotic score remained independent prognostic predictors for the chance to reach the complete remission (P = 0.003 and P = 0.044, respectively) and longer overall survival (OS) (P = 0.002 and P = 0.046, respectively). Significantly, the better response to immunochemotherapy (ICH) in comparison with CH has been achieved in patients with expression of caspase-3, c-FLIP, and survivin and in patients without the immunoexpression of XIAP. In addition, ICH was superior to CH in both bcl-2-positive and bcl-2-negative patients. CONCLUSION: The results of this study showed that the dysregulation of apoptosis can appear on different places of apoptotic cascade in DLBCL. Apoptotic score is a more useful tool in predicting therapy response and OS of patients with DLBCL than single apoptotic parameters and along with IPI could help to identify a high-risk group of newly diagnosed nodal DLBCL.