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Chemical composition, antioxidant capacity, and antimicrobial activity of lavender essential oils (LEOs) extracted from three different varieties of Lavandula angustifolia Mill. (1-Moldoveanca 4, 2-Vis magic 10, and 3-Alba 7) have been determined. These plants previously patented in the Republic of Moldova were cultivated in an organic agriculture system in the northeastern part of Romania and then harvested in 3 consecutive years (2017-2019) to obtain the essential oils. From the inflorescences in the complete flowering stage, the LEOs were extracted by hydrodistillation. Then, their composition was analyzed by gas chromatography coupled with mass spectrometry (GC-MS) and by Fourier Transformed Infrared spectroscopy (FT-IR). The major identified constituents are as follows: linalool (1: 32.19-46.83%; 2: 29.93-30.97%; 3: 31.97-33.77%), linalyl acetate (1: 17.70-35.18%; 2: 27.55-37.13%; 3: 28.03-35.32%), and terpinen-4-ol (1: 3.63-7.70%; 2: 3.06-7.16%; 3: 3.10-6.53%). The antioxidant capacity as determined by ABTS and DPPH assays indicates inhibition, with the highest activity obtained for LEO var. Alba 7 from 2019. The in vitro antimicrobial activities of the LEOs and combinations were investigated as well, by using the disk diffusion method and minimum inhibitory concentration (MIC) against the Gram-positive bacterial strain Staphylococcus aureus (ATCC 6538), Gram-negative Pseudomonas aeruginosa (ATCC 27858), Escherichia coli (ATCC 25922), the yeast Candida albicans (ATCC 10231), and clinical isolates. Our results have shown that LEOs obtained from the three studied varieties of L. angustifolia manifest significant bactericidal effects against tested microorganisms (Staphylococcus aureus and Escherichia coli), and antifungal effects against Candida albicans. The mixture of LEOs (Var. Alba 7) and geranium, respectively, in tea tree EOs, in different ratios, showed a significant enhancement of the antibacterial effect against all the studied strains, except Pseudomonas aeruginosa.
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Antiinfecciosos , Antioxidantes , Bacterias/crecimiento & desarrollo , Candida albicans/crecimiento & desarrollo , Lavandula/química , Aceites Volátiles , Antiinfecciosos/química , Antiinfecciosos/aislamiento & purificación , Antiinfecciosos/farmacología , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Aceites Volátiles/química , Aceites Volátiles/aislamiento & purificación , Aceites Volátiles/farmacologíaRESUMEN
Cannabis sativa L. (hemp) is a plant used in the textile industry and green building material industry, as well as for the phytoremediation of soil, medical treatments, and supplementary food products. The synergistic effect of terpenes, flavonoids, and cannabinoids in hemp extracts may mediate the biogenic synthesis of metal nanoparticles. In this study, the chemical composition of aqueous leaf extracts of three varieties of Romanian hemp (two monoecious, and one dioecious) have been determined by Fourier-Transformed Infrared spectroscopy (FT-IR), high-performance liquid chromatography, and mass spectrometry (UHPLC-DAD-MS). Then, their capability to mediate the green synthesis of silver nanoparticles (AgNPs) and their pottential antibacterial applications were evaluated. The average antioxidant capacity of the extracts had 18.4 ± 3.9% inhibition determined by 2,2-diphenyl-1-picrylhydrazyl (DPPHâ¢) and 78.2 ± 4.1% determined by 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS™) assays. The total polyphenolic content of the extracts was 1642 ± 32 mg gallic acid equivalent (GAE) L-1. After this, these extracts were reacted with an aqueous solution of AgNO3 resulting in AgNPs, which were characterized by UV-VIS spectroscopy, FT-IR, scanning electron microscopy (SEM-EDX), and dynamic light scattering (DLS). The results demonstrated obtaining spherical, stable AgNPs with a diameter of less than 69 nm and an absorbance peak at 435 nm. The mixture of extracts and AgNPs showed a superior antioxidant capacity of 2.3 ± 0.4% inhibition determined by the DPPH⢠assay, 88.5 ± 0.9% inhibition as determined by the ABTSâ¢+ assay, and a good antibacterial activity against several human pathogens: Escherichia coli, Klebsiella pneumoniae, Pseudomonas fluorescens, and Staphylococcus aureus.
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Antibacterianos/química , Cannabis/química , Tecnología Química Verde/métodos , Nanopartículas del Metal/química , Extractos Vegetales/química , Antioxidantes/química , Hojas de la Planta/química , Polifenoles/análisis , Plata/químicaRESUMEN
Harnessing of a branched structure is a novel approach in the design of cell-penetrating peptides and it has provided highly efficient transfection reagents for intracellular delivery of nucleic acids. The new stearylated TP10 analogs, NickFects, condense plasmid DNA, splice correcting oligonucleotides and short interfering RNAs into stable nanoparticles with a size of 62-160nm. Such nanoparticles have a negative surface charge (-11 to -18mV) in serum containing medium and enable highly efficient gene expression, splice correction and gene silencing. One of the novel peptides, NickFect51 is capable of transfecting plasmid DNA into a large variety of cell lines, including refractory suspension and primary cells and in several cases exceeds the transfection level of commercially available reagent Lipofectamine™ 2000 without any cytotoxic side effects. Additionally we demonstrate the advantages of NickFect51 in a protein production system, QMCF technology, for expression and production of recombinant proteins in hardly transfectable suspension cells.
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Péptidos de Penetración Celular/química , Técnicas de Transferencia de Gen , Vectores Genéticos/genética , Ácidos Nucleicos/genética , Secuencia de Aminoácidos , Animales , Células CHO , Línea Celular Tumoral , Membrana Celular/metabolismo , Membrana Celular/ultraestructura , Células Cultivadas , Cricetinae , Cricetulus , Vectores Genéticos/química , Células HEK293 , Células HeLa , Humanos , Células Jurkat , Luciferasas/genética , Microscopía Electrónica de Transmisión , Nanopartículas/química , Nanopartículas/ultraestructura , Oligonucleótidos/genética , Plásmidos/química , Plásmidos/genética , ARN Interferente Pequeño/genética , Ácidos Esteáricos/química , Transfección/métodosRESUMEN
In seeking alternative cancer treatments, antimicrobial peptides (AMPs), sourced from various life forms, emerge as promising contenders. These endogenous peptides, also known as host defense peptides (HDPs), play crucial roles in immune defenses against infections and exhibit potential in combating cancers. With their diverse defensive functions, plant-derived AMPs, such as thionins and defensins, offer a rich repertoire of antimicrobial properties. Insects, amphibians, and animals contribute unique AMPs like cecropins, temporins, and cathelicidins, showcasing broad-spectrum activities against bacteria, fungi, and viruses. Understanding these natural peptides holds significant potential for developing effective and targeted therapies against cancer and infectious diseases. Antimicrobial peptides (AMPs) exhibit diverse structural characteristics, including α-helical, ß-sheet, extended, and loop peptides. Environmental conditions influence their structure, connecting to changes in cell membrane hydrophobicity. AMPs' actions involve direct killing and immune regulation, with additional activities like membrane depolarization. In this review, we focus on antimicrobial peptides that act as anticancer agents and AMPs that exhibit mechanisms akin to antimicrobial activity. Buforin AMPs, particularly Buforin I and II, derived from histone H2A, demonstrate antibacterial and anticancer potential. Buforin IIb and its analogs show promise, with selectivity for cancer cells. Despite the challenges, AMPs offer a unique approach to combat microbial resistance and potential cancer treatment. In various cancer types, including HeLa, breast, lung, ovarian, prostate, and liver cancers, buforins demonstrate inhibitory effects and apoptosis induction. To address limitations like stability and bioavailability, researchers explore buforin-containing bioconjugates, covalently linked with nanoparticles or liposomes. Bioconjugation enhances specificity-controlled release and combats drug resistance, presenting a promising avenue for targeted cancer treatment. Clinical translation awaits further evaluation through in vivo studies and future clinical trials.
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Gliomas are therapeutically challenging cancers with poor patient prognosis. New drug delivery strategies are needed to achieve a more efficient chemotherapy-based approach against brain tumors. The current paper demonstrates development of a tumor-targeted delivery vector that is based on a cell-penetrating peptide pVEC and a novel glioma-targeting peptide sequence gHo. The unique tumor-homing peptide gHo was identified using in vitro phage display technology. The novel delivery vector, which we designated as gHoPe2, was constructed by a covalent conjugation of pVEC, gHo, and a cargo; the latter could be either a labeling moiety (such as a fluorescent marker) or a cytostatic entity. Using a fluorescent marker, we demonstrate efficient uptake of the vector in glioma cells and selective labeling of glioma xenograft tumors in a mouse model. This is the first time that we know where in vitro phage display has yielded an efficient, in vivo working vector. We also demonstrate antitumor efficacy of the delivery vector gHoPe2 using a well-characterized chemotherapeutic drug doxorubicin. Vectorized doxorubicin proved to be more efficient than the free drug in a mouse glioma xenograft model after systemic administration of the drugs. In conclusion, we have characterized a novel glioma-homing peptide gHo, demonstrated development of a new and potential glioma-targeted drug delivery vector gHoPe2, and demonstrated the general feasibility of the current approach for constructing cell-penetrating peptide-based targeted delivery systems.
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Neoplasias Encefálicas/tratamiento farmacológico , Péptidos de Penetración Celular/administración & dosificación , Sistemas de Liberación de Medicamentos/métodos , Vectores Genéticos/administración & dosificación , Glioma/tratamiento farmacológico , Secuencia de Aminoácidos , Animales , Neoplasias Encefálicas/genética , Línea Celular Tumoral , Péptidos de Penetración Celular/genética , Femenino , Vectores Genéticos/genética , Glioma/genética , Células HEK293 , Células HeLa , Humanos , Ratones , Ratones Desnudos , Datos de Secuencia Molecular , Ensayos Antitumor por Modelo de Xenoinjerto/métodosRESUMEN
STUDY QUESTION: Do cell penetrating peptides (CPPs) translocate into spermatozoa and, if so, could they be utilized to deliver a much larger protein cargo? SUMMARY ANSWER: Chemically diverse polycationic CPPs rapidly and efficiently translocate into spermatozoa. They exhibit differential accumulation within intracellular compartments without detrimental influences upon cellular viability or motility but they are relatively ineffective in transporting larger proteins. WHAT IS ALREADY KNOWN: Endocytosis, the prevalent route of protein internalization into eukaryotic cells, is severely compromised in mature spermatozoa. Thus, the translocation of many bioactive agents into sperm is relatively inefficient. However, the delivery of bioactive moieties into mature spermatozoa could be significantly improved by the identification and utility of an efficient and inert vectorial delivery technology. STUDY DESIGN: CPP translocation efficacies, their subsequent differential intracellular distribution and the influence of peptides upon viability were determined in bovine spermatozoa. Temporal analyses of sperm motility in the presence of exogenously CPPs utilized normozoospermic human donor samples. MATERIALS AND METHODS: CPPs were prepared by manual, automated and microwave-enhanced solid phase synthesis. Confocal fluorescence microscopy determined the intracellular distribution of rhodamine-conjugated CPPs in spermatozoa. Quantitative uptake and kinetic analyses compared the translocation efficacies of chemically diverse CPPs and conjugates of biotinylated CPPs and avidin. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) conversion assays were employed to analyse the influence of CPPs upon sperm cell viability and sperm class assays determined the impact of CPPs on motility in capacitated and non-capacitated human samples. MAIN RESULTS: Chemically heterogeneous CPPs readily translocated into sperm to accumulate within discrete intracellular compartments. Mitoparan (INLKKLAKL(Aib)KKIL), for example, specifically accumulated within the mitochondria located in the sperm midpiece. The unique plasma membrane composition of sperm is a critical factor that directly influences the uptake efficacy of structurally diverse CPPs. No correlations in efficacies were observed when comparing CPP uptake into sperm with either uptake into fibroblasts or direct translocation across a phosphatidylcholine membrane. These comparative investigations identified C105Y (CSIPPEVKFNKPFVYLI) as a most efficient pharmacokinetic modifier for general applications in sperm biology. Significantly, CPP uptake induced no detrimental influence upon either bovine sperm viability or the motility of human sperm. As a consequence of the lack of endocytotic machinery, the CPP-mediated delivery of much larger protein complexes into sperm is relatively inefficient when compared with the similar process in fibroblasts. LIMITATIONS, REASONS FOR CAUTION: It is possible that some CPPs could directly influence aspects of sperm biology and physiology that were not analysed in this study. WIDER IMPLICATIONS OF THE FINDINGS: CPP technologies have significant potential to deliver selected bioactive moieties and so could modulate the biology and physiology of human sperm biology both prior- and post-fertilization.
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Péptidos de Penetración Celular/metabolismo , Motilidad Espermática , Espermatozoides/metabolismo , Animales , Bovinos , Permeabilidad de la Membrana Celular , Endocitosis , Humanos , Masculino , Ratones , Células 3T3 NIH , Transporte de ProteínasRESUMEN
Membrane-active peptides (MAPs) possess unique properties that make them valuable tools for studying membrane structure and function and promising candidates for therapeutic applications. This review paper provides an overview of the fundamental aspects of MAPs, focusing on their membrane interaction mechanisms and potential applications. MAPs exhibit various structural features, including amphipathic structures and specific amino acid residues, enabling selective interaction with multiple membranes. Their mechanisms of action involve disrupting lipid bilayers through different pathways, depending on peptide properties and membrane composition. The therapeutic potential of MAPs is significant. They have demonstrated antimicrobial activity against bacteria and fungi, making them promising alternatives to conventional antibiotics. MAPs can selectively target cancer cells and induce apoptosis, opening new avenues in cancer therapeutics. Additionally, MAPs serve as drug delivery vectors, facilitating the transport of therapeutic cargoes across cell membranes. They represent a fascinating class of biomolecules with significant potential in basic research and clinical applications. Understanding their mechanisms of action and designing peptides with enhanced selectivity and efficacy will further expand their utility in diverse fields. Exploring MAPs holds promise for developing novel therapeutic strategies against infections, cancer, and drug delivery challenges.
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Elevated carbon dioxide and drought are significant stressors in light of climate change. This study explores the interplay between elevated atmospheric CO2, drought stress, and plant physiological responses. Two Brassica oleracea varieties (cauliflowers and cabbage) were utilized as model plants. Our findings indicate that elevated CO2 accelerates assimilation rate decline during drought. The integrity of photosynthetic components influenced electron transport, potentially due to drought-induced nitrate reductase activation changes. While CO2 positively influenced photosynthesis and water-use efficiency during drought, recovery saw decreased stomatal conductance in high-CO2-grown plants. Drought-induced monoterpene emissions varied, influenced by CO2 concentration and species-specific responses. Drought generally increased polyphenols, with an opposing effect under elevated CO2. Flavonoid concentrations fluctuated with drought and CO2 levels, while chlorophyll responses were complex, with high CO2 amplifying drought's effects on chlorophyll content. These findings contribute to a nuanced understanding of CO2-drought interactions and their intricate effects on plant physiology.
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Finding suitable nonviral delivery vehicles for nucleic acid-based therapeutics is a landmark goal in gene therapy. Cell-penetrating peptides (CPPs) are one class of delivery vectors that has been exploited for this purpose. However, since CPPs use endocytosis to enter cells, a large fraction of peptides remain trapped in endosomes. We have previously reported that stearylation of amphipathic CPPs, such as transportan 10 (TP10), dramatically increases transfection of oligonucleotides in vitro partially by promoting endosomal escape. Therefore, we aimed to evaluate whether stearyl-TP10 could be used for the delivery of plasmids as well. Our results demonstrate that stearyl-TP10 forms stable nanoparticles with plasmids that efficiently enter different cell-types in a ubiquitous manner, including primary cells, resulting in significantly higher gene expression levels than when using stearyl-Arg9 or unmodified CPPs. In fact, the transfection efficacy of stearyl-TP10 almost reached the levels of Lipofectamine 2000 (LF2000), however, without any of the observed lipofection-associated toxicities. Most importantly, stearyl-TP10/plasmid nanoparticles are nonimmunogenic, mediate efficient gene delivery in vivo, when administrated intramuscularly (i.m.) or intradermally (i.d.) without any associated toxicity in mice.
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Péptidos de Penetración Celular/metabolismo , Técnicas de Transferencia de Gen , Vectores Genéticos , Plásmidos/metabolismo , Transfección/métodos , Animales , Transporte Biológico , Línea Celular , Cricetinae , Cricetulus , Portadores de Fármacos , Sistemas de Liberación de Medicamentos , Endosomas/metabolismo , Terapia Genética/métodos , Humanos , Ratones , Ratones Endogámicos BALB C , Ácidos Nucleicos/metabolismoRESUMEN
Climate change will determine a sharp increase in carbon dioxide in the following years. To study the influence of elevated carbon dioxide on plants, we grew 13 different species and varieties from the Brassicaceae family at three carbon dioxide concentrations: 400, 800, and 1200 ppmv. The photosynthetic parameters (assimilation rate and stomatal conductance to water vapor) increase for all species. The emission of monoterpenes increases for plants grown at elevated carbon dioxide while the total polyphenols and flavonoids content decrease. The chlorophyll content is affected only for some species (such as Lipidium sativum), while the ß-carotene concentrations in the leaves were not affected by carbon dioxide.
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The purpose of this study is to reveal the chemical and biochemical characteristics and the potential aromatherapy applications of the essential oil (EO) of Salvia officinalis (common sage) within a hospital environment. The chemical composition was determined by gas chromatography with mass spectrometry and ATR-FTIR spectroscopy. Three types of sage EOs were included in this study: two commercial oils and one oil obtained by in-house hydrodistillation. Based on the findings, these EOs were included in different chemotypes. The first two samples were similar to the most common chemotype (α-thujone > camphor > 1,8-cineole > ß-thujone), while the in-house sage EO revealed a high content of 1,8-cineole, borneol, α-thujone, similar to the Dalmatian type. The latter sample was selected to be evaluated for its antioxidant and medical effects, as borneol, a bicyclic monoterpene, is known as a substance with anesthetic and analgesic effects in traditional Asian medicine. The study suggests that the antioxidant capacity of the sage EO is modest (33.61% and 84.50% inhibition was determined by DPPH and ABTS assays, respectively), but also that the inhalation of sage EO with high borneol content by hospitalized patients could improve these patients' satisfaction.
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Cell-penetrating peptide based vehicles have been developed for the delivery of different payloads into the cells in culture and in animals. However, several biological features, among which is the tendency to trigger innate immune response, limit the development of highly efficient peptide-based drug delivery vectors. This study aims to evaluate the influence of transportan 10 (TP10) and its chemically modified derivatives, PepFects (PFs), on the innate immune response of the host system. PFs have shown high efficiency in nucleic acid delivery in vitro and in vivo; hence, the estimation of their possible toxic side effects would be of particular interest. In this study, we analyzed cytotoxic and immunogenic response of PF3, PF4, and PF6 peptides in monocytic leukemia and peripheral blood mononuclear cell lines. In comparison with amphipathic PFs, TP10, TAT, stearyl-(RxR)(4) peptides, and the most widely used transfection reagents Lipofectamine 2000 and Lipofectamine RNAiMAX were also analyzed in this study. IL-1ß, IL-18, and TNF-α cytokine release was detected using highly sensitive enzyme-linked immunosorbent assay (ELISA). Cell viability was detected by measuring the activity of cellular enzymes that reduce water-soluble tetrazolium salts to formazan dyes and apoptosis was evaluated by measuring the levels of caspase-1 and caspase-3/7 over untreated cells. All peptides were found to be nontoxic and nonimmunogenic in vitro at the concentrations of 10 µM and 5 µM, respectively, and at a dose of 5 mg/kg in vivo, suggesting that these CPPs exhibit a promising potential in the delivery of therapeutic molecules into the cell without risks of toxicity and inflammatory reactions.
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Péptidos de Penetración Celular/inmunología , Péptidos de Penetración Celular/toxicidad , Portadores de Fármacos/toxicidad , Galanina/inmunología , Galanina/toxicidad , Proteínas Recombinantes de Fusión/inmunología , Proteínas Recombinantes de Fusión/toxicidad , Venenos de Avispas/inmunología , Venenos de Avispas/toxicidad , Animales , Caspasas/metabolismo , Línea Celular Tumoral/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Péptidos de Penetración Celular/química , Péptidos de Penetración Celular/genética , Células Cultivadas , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Galanina/química , Humanos , Interleucina-18/inmunología , Interleucina-1beta/inmunología , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Lipopéptidos/química , Lipopéptidos/inmunología , Lipopéptidos/toxicidad , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas Recombinantes de Fusión/química , Transfección , Factor de Necrosis Tumoral alfa/inmunología , Venenos de Avispas/químicaRESUMEN
Many plants belonging to the Lamiaceae family are rich in essential oils (EOs) which are intensively used for aromatherapy, food and beverage flavoring, alternative medicine, cosmetics, and perfumery. Aerial parts of Thymus vulgaris L., Thymus pannonicus All., Lavandula angustifolia L., Lavandula x intermedia, Origanum vulgare L., and Origanum vulgare var. aureum L. were subjected to hydrodistillation, and both resulting fractions were analyzed. The purpose of this study was to determine the chemical composition, antioxidant activity, and total phenolic content of six essential oils and their corresponding hydrolats (HDs) through GC-MS and spectrophotometric analyses. Overall, 161 compounds were identified, some found exclusively in essential oils and others in hydrolats, making them individual products with specific end purposes. The total phenolic content was the highest for the Thymus vulgaris L. EOs (3022 ± mg GAE L-1), because of its high phenolic oxygenated monoterpenes content (thymol and carvacrol) and the smallest for the Lavandula angustifolia L. EOs (258.31 ± 44.29 mg GAE L-1), while hydrolats varied from 183.85 ± 0.22 mg GAE L-1 for Thymus vulgaris L. HD and 7.73 mg GAE L-1 for Thymus pannonicus All. HD. Significant antioxidant effects determined through DPPH⢠and ABTSâ¢+ assays were also observed in samples with higher hydrophilic compounds. The highest antioxidant activity was determined for Thymus vulgaris L. EO and its corresponding HD. Although EOs are the principal traded economic product, HDs represent a valuable by-product that could still present intense antiseptic activities, similar to their corresponding EOs (thyme and oregano), or have multiple aromatherapy, cosmetics, and household applications (lavender and lavandin).
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The purpose of this study was to analyze the chemical composition and antimicrobial activity of some thymus populations collected from five different locations in Western Romania. The chemical compositions of the essential oils (EOs) were studied through GC-MS, and the biological activities were evaluated using the microdilution method. The EO yield ranged between 0.44% and 0.81%. Overall, 60 chemical compounds were identified belonging to three chemotypes: thymol (three populations), geraniol (one population) and carvacrol (one population). Thymus vulgaris L. is distinguished by a high content of thymol, while species of spontaneous flora (Th. odoratissimus and Th. pulegioides) contain, in addition to thymol, appreciable amounts of carvacrol and geraniol. The antimicrobial activity of each the five oils was tested on Staphylococcus aureus (ATCC 25923), Streptococcus pyogenes (ATCC 19615), Esherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Shigella flexneri (ATCC 12022), Salmonella typhimurium (ATCC 14028), Haemophilus influenzae type B (ATCC 10211), Candida albicans (ATCC 10231) and Candida parapsilopsis (ATCC 22019). The EOs showed biological activity on Gram-positive/Gram-negative/fungal pathogens, the most sensitive strains proving to be S. pyogenes, S. flexneri, S. typhimurium and C. parapsilopsis with an MIC starting at 2 µL EO/100 µL. The species sensitive to the action of Thymus sp. from culture or spontaneous flora are generally the same, but it should be noted that T. odoratissimus has a positive inhibition rate higher than other investigated EOs, regardless of the administered oil concentration. To date, there is no research work presenting the chemical and antimicrobial profiling of T. odoratissimus and the correlations between the antimicrobial potential and chemical composition of wild and cultivated populations of thyme (Thymus sp.) growing in Western Romania.
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Satureja hortensis is one of the representative plants from the Lamiaceae family, and its essential oil has been used in various applicative fields, from the food industry to aromatherapy. The changes that occur in heated samples at different temperatures (160, 175, 190 ºC) over different periods (0.5 and 2.5 h) in Satureja hortensis essential oil composition and chemical properties were evaluated. The results showed that the major chemical composition constituents of the investigated essential oil are γ-terpinene + α-terpinolene and carvacrol + p-cymene and the thermal behavior is dependent on the content. This composition drastically changes through the heating of the samples and causes significant changes in thermal behavior. The present study demonstrated that the concentration of carvacrol in S. hortensis essential oil is increasing after heating treatment, and the sample heated at 190 ºC for 2.5 h contained more than 91% carvacrol. This simple treatment is a rapid way to obtain carvacrol from the essential oil that has high potential as a natural preservative suitable for the food industry and alternative and complementary medicine.
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The aim of the present study was to establish the influence of high-temperature heating on the composition and thermal behavior of coffee oils obtained from Arabica green and roasted coffee beans, respectively. Morphological studies performed using scanning electron microscopy revealed the oil bodies uniformly distributed within the cells in both types of coffee beans analyzed. The obtained oils have a fatty acid composition rich in linoleic acid, palmitic acid, oleic acid, stearic acid, arachidic acid and linolenic acid. The total content of saturated fatty acids of investigated oils was 49.38 and 46.55%, the others being unsaturated fatty acids. The thermal behavior and thermo-oxidative stability of coffee oils extracted from green coffee beans and roasted coffee beans, the coffee oil high-temperature heated up to 200 °C, were investigated using simultaneous thermal analysis TG/DTG/DTA, in an oxidizing atmosphere. The data obtained for the analyzed samples depend mainly on the nature and compositions of fatty acids, and to a lesser extent on the roasting process of the coffee beans and the high-temperature heating process of the extracted oil. The chromatographic and TG/DTG/DTA data suggest that Arabica coffee oil has great potential for use in technological processes which require high-temperature heating (e.g. food industry or pastries).
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Café/química , Extractos Vegetales/química , Aceites de Plantas/química , Café/ultraestructura , Ácidos Grasos/análisis , Cromatografía de Gases y Espectrometría de Masas , Calor , Microscopía Electrónica de Rastreo , Oxidación-Reducción , Semillas/química , Semillas/ultraestructuraRESUMEN
The intrinsic property of cell-penetrating peptides (CPPs) to deliver therapeutic molecules (nucleic acids, drugs, imaging agents) to cells and tissues in a nontoxic manner has indicated that they may be potential components of future drugs and disease diagnostic agents. These versatile peptides are simple to synthesize, functionalize, and characterize yet are able to deliver covalently or noncovalently conjugated bioactive cargos (from small chemical drugs to large plasmid DNA) inside cells, primarily via endocytosis, in order to obtain high levels of gene expression, gene silencing, or tumor targeting. Typically, CPPs are often passive and nonselective yet must be functionalized or chemically modified to create effective delivery vectors that succeed in targeting specific cells or tissues. Furthermore, the design of clinically effective systemic delivery systems requires the same amount of attention to detail in both design of the delivered cargo and the cell-penetrating peptide used to deliver it.
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Péptidos de Penetración Celular/síntesis química , Péptidos de Penetración Celular/metabolismo , Portadores de Fármacos/síntesis química , Portadores de Fármacos/metabolismo , Diseño de Fármacos , Secuencia de Aminoácidos , Animales , Péptidos de Penetración Celular/química , Portadores de Fármacos/química , Humanos , Datos de Secuencia Molecular , Nanopartículas/química , Nanopartículas/metabolismo , Transporte de ProteínasRESUMEN
In recent years, oligonucleotide-based molecules have been intensely used to modulate gene expression. All these molecules share the common feature of being essentially impermeable over cellular membranes and they therefore require efficient delivery vectors. Cell-penetrating peptides are a group of delivery peptides that has been readily used for nucleic acid delivery. In particular, polyarginine and derivates thereof, i.e. the (RxR)(4) peptide, have been applied with success both in vitro and in vivo. A major problem, however, with these arginine-rich peptides is that they frequently remain trapped in endosomal compartments following internalization. The activity of polyarginine has previously been improved by conjugation to a stearyl moiety. Therefore, we sought to investigate what impact such modification would have on the pre-clinically used (RxR)(4) peptide for non-covalent delivery of plasmids and splice-correcting oligonucleotides (SCOs) and compare it with stearylated Arg9 and Lipofectamine 2000. We show that stearyl-(RxR)(4) mediates efficient plasmid transfections in several cell lines and the expression levels are significantly higher than when using unmodified (RxR)(4) or stearylated Arg9. Although the transfection efficiency is lower than with Lipofectamine 2000, we show that stearyl-(RxR)(4) is substantially less toxic. Furthermore, using a functional splice-correction assay, we show that stearyl-(RxR)(4) complexed with 2'-OMe SCOs promotes significant splice correction whereas stearyl-Arg9 fails to do so. Moreover, stearyl-(RxR)(4) promotes dose-dependent splice correction in parity with (RxR)(4)-PMO covalent conjugates, but at least 10-times lower concentration. These features make this stearic acid modified analog of (RxR)(4) an intriguing vector for future in vivo experiments.