A scaris is a genus of parasitic worms (helminths) found in the small intestine of various mammalian hosts, including Ascaris lumbricoides in humans, Parascaris equorum and P univalens in horses, Ascaris suum in pigs, Toxocara cati in cats and Toxocara canis in dogs. To date, Parascaris spp. are the only Ascaris worms that have developed resistance to anthelmintic drugs. The mechanisms of resistance in Parascaris spp are incompletely understood, partly due to the absence of robust in-vitro models. Further complicating in-vitro studies, Parascaris spp lack a free-living larval stage as their larva only hatch within the host. The aim of this study was to develop in-vitro methods for hatching, scoring the viability of Parascaris L3 larvae and exposing them to the anthelmintic drugs ivermectin, pyrantel, thiabendazole, and the herbal extract carvacrol. This study shows that mechanical Ascaris egg breaking using a homogenizer resulted in a hatching rate of 98%. Our viability scoring system could distinguish an ivermectin resistant larvae from an ivermectin susceptible larvae derived from different farms. This indicates that this method may have utility for the screening of larvae ivermectin resistance on the level of farm populations. Interestingly, a highly paralytic effect observed after carvacrol exposure. Carvacrol shows direct paralytic effects on Parascaris larvae in a dose-dependent manner, as higher concentrations were lethal to all exposed larvae. This result presents a potential future opportunity for carvacrol used in the treatment of Ascaris infections. To conclude our results, we have successfully developed an in-vitro model as well as a scoring system for the viability of Parascaris L3 stage larvae, which can be used for assaying the effect on larvae after drug exposure

Human norovirus (HuNoVs) are the most common enteric pathogen around the world that cause ~50% of foodborne illness of disease outbreaks annually. HuNoVs are the member of the Caliciviridae family, which consist of small (38 nm), unenveloped, single stranded RNA (ssRNA) viruses. Norovirus are divided into 5 genogroup (GI, GII, GIII, GIV, GV, GVI and GVII). The GI, GII, and GIV cause human illness, in addition, GII.4 genotype cause the most human disease. Due to HuNoVs are difficult cultured in vitro, the cultivable HuNoVs surrogates have been widely studied. Recently, some studies have been conducted with HuNoVs surrogates, for example bacteriophage MS2. MS2 is conservative surrogate for nonenveloped viruses which there is a close relationship to the behavior of HuNoVs, thus we can examine the infection control measures for HuNoVs. Despite plenty of treatment method been done on testing antiviral effect on bacteriophage MS2, for example UV inactivation, steam ultrasound and antimicrobial etc., plant-based nanoemulsion treatment has yet to be explored. Carvacrol is a major component of oregano essential oil and is responsible for their antimicrobial activity on the growth of various microorganism. In this study, carvacrol nanoemulsions were formed by using the spontaneous emulsification for testing the nanoemulsion stability (14 days shelf life study on its droplet size and particle charge) and antimicrobial activity. In carvacrol nanoemulsion 14 days shelf life test, the droplet size and particle charge stay stable at three different treatment environments (4°C, 20°C and 37°C). The results proved that nanoemulsion (was formed with surfactant agents and medium-chain triglycerides) is stable system that gives consistent droplet size and charge. Although, the low antimicrobial activity was investigated at carvacrol nanoemulsion, the strong antimicrobial effects have been found when carvacrol or carvacrol combined with ionic surfactant of treatment on MS2 and Escherichia coli. Taken together, in the wake of growing consumer demand for different “natural” products in a number of industries, our study broadly informs the development and study of functionalized carvacrol active compound that can not only provide beneficial health for human but can also examine antimicrobial efficacy of control measures for public health.

No description available.

<p>U okviru ove disertacije su ispitane mogućnosti primene savremenih postupaka za dobijanje ekstrakata rtanjskog čaja (Satureja montana L.).<br />U ekstraktima dobijenim primenom superkritičnog ugljendioksida (različitih gustina) utvrđeno je prisustvo karvakrola kao najzastupljenije komponente. Ekstrakt sa najvećim sadržajem karvakrola je dobijen pri uslovima 350 bar i 50 &deg;C (60,82%), dok je najveći prinos karvakrola ostvaren pri pritiscima 325 i 350 bar i temperaturi 60 &deg;C, i iznosio je 2,4 g/100 g droge. Primenom ugljendioksida na većim pritiscima se ostvaruje efikasnija ekstrakcija karvakrola iz droge, kao i proizvodnja ekstrakata sa većim sadržajima karvakrola. Druge komponente prisutne u ekstraktima u znatno nižim koncentracijama su: p-cimen, borneol, trans-kariofilen, kariofilen-oksid, &gamma;-terpinen i linalool.<br />Da bi se utvrdila mogućnost unapređenja prinosa ekstrakcije superkritičnim ugljendioksidom, kao i pobolj&scaron;anja kvaliteta ekstrakata, ispitan je uticaj različitih predtretmana biljnog materijala. Ustanovljeno je da predtretman vodom najznačajnije povećava prinos ekstrakcije (za 25%), dok je ultrazvučni predtretman najadekvatniji izbor za dobijanje ekstrakata sa najvećim sadržajem karvakrola (66,46%). Međutim, najveći prinos ekstrakcije karvakrola iz 100 g droge postignut je primenom etanola i vode kao predtretmana.<br />Ekstrakti dobijeni primenom subkritične vode (uz variranje temperature i vremena ekstrakcije) su hemijski okarakterisani i radi utvrđivanja uslova ekstrakcije pri kojima se postiže najveći kvalitet ekstrakata u pogledu sadržaja polifenolnih komponenti, kao i antioksidantne aktivnosti, primenjena je metoda odzivne povr&scaron;ine (RSM). Utvrđena je prednost primene subkritične vode u odnosu na klasičnu metodu ekstrakcije u pogledu sadržaja polifenolnih komponenti i antioksidantne aktivnosti.<br />Suvi ekstrakti S. montana dobijeni spray drying tehnologijom su&scaron;enja (sa različitim koncentracijama maltodekstrina) su ispitani u pogledu fizičkih i hemijskih osobina. Takođe, radi ispitivanja mogućnosti primene u vidu funkcionalnih ili prehrambenih proizvoda, izvr&scaron;ena je senzorna analiza dobijenih suvih ekstrakata, i analizirana je njihova farmakolo&scaron;ka aktivnost u smislu ACE inhibitornog i antioksidantnog delovanja.<br />Ustanovljeno je da je najniža koncentracija maltodekstrina (10%) najadekvatnija za dobijanje suvog ekstrakta S. montana sa najvećim sadržajem polifenolnih komponenti, sadržajem etarskog ulja i u pogledu antioksidantne i ACE inhibitorne aktivnosti. Takođe, suvi ekstrakt sa 10% maltodekstrina ispunjava i ostale fizičko-hemijske parametre, kojima se osigurava kako efikasno su&scaron;enje, tako i kvalitet ekstrakta.<br />In vivo aktivnost odabranih ekstrakata S. montana ispitana je na modelu hepatotoksičnosti indukovane primenom ugljentetrahlorida. Analiziran je uticaj ekstrakata na funkciju jetre, enzimske parametre oksidativnog stresa i serumske parametre oksidativnog o&scaron;tećenja izazvanog ugljentetrahloridom i ustanovljen je hepatoprotektivni učinak ekstrakata S. montana. Takođe, ispitano je dejstvo ekstrakata na rast Ehrlich-ovih ascitnih tumora implantiranih mi&scaron;evima, kao i na biohemijske parametre oksidativnog stresa u ćelijama tumora i utvrđeno je da je vreme aplikacije ekstrakata od izuzetnog značaja.</p> / <p>This dissertation investigates the possibilities of applying contemporary processes for obtaining extracts of winter savory (Satureja montana L.).<br />Carvacrol was determined to be the most abundant component in extracts acquired by using supercritical carbon dioxide (of various densities). Extract with the richest content of carvacrol was obtained under conditions of 350 bar and 50 &deg;C (60,82%), while the highest carvacrol yield was produced under 325 bar and 350 bar pressures and at a 60 &deg;C temperature reaching 2.4 g/100 g of dry weight. By applying carbon dioxide on higher pressures, a more efficient carvacrol extraction from herbal material is performed, alongside production of extracts with higher carvacrol content. Other components in the extracts present in significantly lower concentrations are the following: p-cymen, borneol, trans-caryophyllene, caryophyllene-oxide, &gamma;-terpinene and linalool.<br />In order to recognize the possibility of improving the extraction yield produced by employing supercritical carbon dioxide and enhancing the quality of extracts, the influence of several different pre-treatments was investigated. It was determined that the water pre-treatment raises the extraction yield the most (for 25%), while the ultrasonic pre-treatment was the most adequate choice for acquiring extracts with the highest content of carvacrol (66.46%). However, the highest carvacrol extraction yield from 100 g of dry weight was achieved by applying ethanol and water as pre-treatment.<br />Extracts which were attained with subcritical water (by varying the temperature and time of extraction) were chemically characterized and Response Surface Method (RSM) was applied for the purposes of determining conditions of extractions under which the highest quality of extracts in terms of content of polyphenolic compounds and antioxidant activity is reached. The advantage of employing subcritical water over the classical method of extraction in respect of content of polyphenolic components and antioxidant activity was demonstrated.<br />Dry extracts of S. montana obtained by using the spray drying technology (with different concentrations of maltodextrine) were examined in terms of their physical and chemical characteristics. Also, in order to investigate the possibility of their use as functional or food products, sensory analysis of attained extracts was performed, and furthermore, their pharmacological activity in terms of the ACE inhibitory and antioxidant effect was analysed as well.<br />It was determined that the lowest concentration of maltodextrin (10%) was the most adequate for obtaining dry extracts of S. montana with the richest content of polyphenolic components and essential oil, and in respect of antioxidant and ACE inhibitory activities. Also, dry extract with 10% of maltodextrin meets all other physical-chemical parameters, which provide efficient drying and quality of extracts.<br />In vivo activity of selected extracts of S. montana was examined on a model of hepatotoxicity induced by applying carbon tetrachloride. The influence of extracts on liver function, enzyme parameters of oxidative stress, and serum parameters of oxidative damage caused by carbon tetrachloride was analysed and hepatoprotective effect of S. montana extracts was determined. In addition, the influence of extracts on the growth of Ehrlich ascite tumors implanted in mice was investigated, and their impact on biochemical parameters of oxidative stress in tumor cells as well, and it was determined that the time of application of extracts is of great significance.</p>

[...]Ce travail de thèse porte sur la conception et la réalisation d’emballages actifs, pour desapplications dans le domaine de l’agroalimentaire. Elle s’inscrit dans le cadre du projetEMAC du groupe Vitagora, dont les objectifs sont d’optimiser la conservation des qualitésorganoleptiques des produits, et d’augmenter la DLC et la DLUO des aliments, à travers lamaîtrise de l’atmosphère des emballages.Dans une première partie, la thèse se concentre sur les aspects matériaux, avec un état de l’artsur la mise en oeuvre des emballages dits « actifs », l’étude du comportement à la mise enoeuvre des mélanges et la réalisation des emballages actifs. Les travaux de cette thèse se sontorientés sur l'amélioration d'emballages existant par l’incorporation directe de moléculesactives (thymol et carvacrol) et de charges végétales (chanvre et cellulose) susceptibles demoduler le comportement des agents actifs introduits dans l'emballageDans un deuxième temps, la thèse fait porter l’analyse sur les propriétés physico-chimiquesdes matériaux. Cette étude permettra de vérifier l’intégrité physique des matériaux, d’analyserle comportement rhéologique et la stabilité thermique au cours du procédé d’élaboration. Endépit des nombreuses contraintes thermo-mécaniques subies par les matériaux à la mise enoeuvre, les agents actifs ne semblent pas subir d’altérations chimiques. La présence de fibrespermet de réduire le taux de perte en agent actif ce qui pourrait être expliqué par leur systèmecapillaire qui sert de réservoir aux agents actifs lors de la mise en oeuvre.La troisième partie porte sur l’étude des interactions contenant/contenu et sur les différentsaspects règlementaires en termes de migration auxquelles sont soumis les emballagesalimentaires. Dans certains cas, la migration globale est supérieure à la LMG de 10 mg/dm².Nous avons optimisé la mise en oeuvre des matériaux en modifiant la réalisation descompounds et en réduisant le taux de fibres de 2,5% à 1% dans les emballages finaux. Unenette amélioration des valeurs de migration globale est observée, on passe ainsi d’une valeurmaximale 10,81 mg/dm² de à 2,85 mg/dm². [...] Enfin, dans la dernière partie, nous abordons l’efficacité de ces emballages actifs vis-à-vis dessouches bactériennes et leur impact sur la conservation des produits de l’industrieagroalimentaire. La capacité antimicrobienne du thymol et du carvacrol reste conservée aprèsla mise en oeuvre, de l’extrusion du mélange maître à la production de pots en injection. Lesplus grandes valeurs d’inhibition sont observées pour les échantillons chargés en fibresvégétales. De plus, il n’est pas nécessaire d’utiliser de fortes concentrations en agents actifspour observer une action sur le développement microbien. Des concentrations minimalesinhibitrices de l’ordre de 10-5 à 10-4 mg d’agent actif par millilitre d’air, ont été mesurées dansle cas des souches bactériennes P. camemberti, B. subtilis, L. plantarum, E. coli. Pour uneincorporation d’environ 0,2 % massique d’agent actif dans les emballages, il est déjà possibled’atteindre une concentration en phase vapeur moyenne de 4x10-5 mg/ml.Les essais préliminaires sur les propriétés sensorielles et organoleptiques montrent que ladurée de conservation d'aliments dans les emballages actifs augmente. Cependant il est aussipossible de noter un impact organoleptique se traduisant dans certains cas par unemodification de la saveur des aliments, dans d’autres cas par une odeur à l’ouverture desemballages. Les meilleurs résultats de conservation des aliments ont été obtenus avec lesfilms co-extrudés, dont la concentration en agent actif est cinq fois plus faible que dans celledes pots réalisés en injection / No abstract

Amebic keratitis and granulomatous amebic encephalitis are diseases caused by free-living amoebas of the Acanthamoeba type. In the case of keratitis, the great problem is the relapse of this disease because of its resistance to medicines used for it, especially the cystic form of the organism. Plant essential oils have been used as potentially active agents against this protist. Therefore, the objective of this study was to determine the amebacide activity of plant essential oils of Lippia type against Acanthamoeba polyphaga trophozoites. To do this, 8 x 104 trophozoites were exposed to increased concentrations of essential oils of Lippia sidoides, Lippia gracilis, Lippia alba and Lippia pedunculosa during 24 hours, as well as of their majority compounds rotundifolona, carvone and carvacrol. Practically all the oil and compound concentrations presented amebacide activity. Based on these results, the researched oils in this study are potential candidates for a complementary healing for amebic keratitis and granulomatous amebic encephalitis. / A ceratite amebiana e a encefalite amebiana granulomatosa são doenças causadas por amebas de vida livre do gênero Acanthamoeba. No caso da ceratite, o grande problema é a recidiva da doença devido à resistência, especialmente da forma cística do organismo, aos medicamentos utilizados. Óleos essenciais de plantas têm sido utilizados como agentes potencialmente ativos contra este protista. Portanto, o objetivo deste estudo foi determinar a atividade amebicida de óleos essenciais de plantas do gênero Lippia frente à trofozoítos de Acanthamoeba polyphaga. Para isto 8 x 104 trofozoítos foram expostos durante 24 horas a concentrações crescentes dos óleos essenciais de Lippia sidoides, Lippia gracilis, Lippia alba e Lippia pedunculosa, bem como de seus compostos majoritários rotundifolona, carvona e carvacrol. Praticamente todas as concentrações dos óleos e compostos apresentaram atividade amebicida. Diante destes resultados, os óleos pesquisados neste estudo são candidatos em potencial para a terapêutica complementar na ceratite amebiana e encefalite amebiana granulomatosa.

No description available.

Carvacrol is a phenol monoterpene found in aromatic plants specially in Lamiaceae family, which has been evaluated in an experimental model of breast cancer. However, any proposed mechanism based on its antitumor effect has not been reported. In our previous study, carvacrol showed a protective effect on 7,12-dimethylbenz[α]anthracene- (DMBA-) induced breast cancer in female rats. The main objective in this research was to evaluate by using in silico study the carvacrol on HER2, PI3Kα, mTOR, hERα, PR, and EGFR receptors involved in breast cancer progression by docking analysis, molecular dynamic, and drug-likeness evaluation. A multilevel computational study to evaluate the antitumor potential of carvacrol focusing on the main targets involved in the breast cancer was carried out. The in silico study starts with protein-ligand docking of carvacrol followed by ligand pathway calculations, molecular dynamic simulations, and molecular mechanics energies combined with the Poisson–Boltzmann (MM/PBSA) calculation of the free energy of binding for carvacrol. As result, the in silico study led to the identification of carvacrol with strong binding affinity on mTOR receptor. Additionally, in silico drug-likeness index for carvacrol showed a good predicted therapeutic profile of druggability. Our findings suggest that mTOR signaling pathway could be responsible for its preventive effect in the breast cancer. / Revisión por pares

Made available in DSpace on 2015-05-14T13:00:13Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2275378 bytes, checksum: fcb4196c739d83997818dc074daf738f (MD5) Previous issue date: 2010-03-11 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The pharmacological effects of carvacrol, a monoterpenoid phenol, on the cardiovascular system were studied in normotensive rats, using in vivo and in vitro techniques. In superior mesenteric artery rings isolated from rats with functional endothelium carvacrol (10-8 - 3 ₓ 10-4 M) concentration-dependently relaxed phenylephrine-induced contractions (pD2 = 4.59  0.02, MR = 103.03  1.5%, N=8) and this effect was not altered after removal of the endothelium (pD2 = 4.36  0.02, MR = 111.03  4.8%, N=8), suggesting that the vasorelaxant response induced by carvacrol appears to be independent of vascular endothelium. Furthermore, carvacrol antagonized the vasoconstriction induced by high K+ solution (Tyrode with 80 mM of KCl) (pD2 = 4.12  0.01, MR = 94.38  3.97%, N=6), inhibited contraction elicited by CaCl2 in depolarizing (KCL 60 mM) nominally without Ca2+ medium out carvacrol also antagonized the contractions induced by the L-type Ca2+ channel agonist, S(-)-Bay K 8644 (pD2 = 4.537  0.023, MR = 9.8  3.58%, N=6), indicating that the vasodilatation involve probably the inhibition of Ca2+ influx through L-type voltage-dependent calcium channels (Cav type-L). Additionally, carvacrol antagonized the contractions induced by CaCl2 in nominally without Ca2+ medium in the presence of PHE and nifedipine, suggesting a possible inhibition of calcium influx by store operated channels (SOC), receptor operated channels (ROC) and/or TRP channels. Interestingly, in a depolarizing (KCL 60 mM) nominally without Ca2+ medium and in the presence of nifedipine, carvacrol also inhibited the contraction induced by CaCl2, suggesting a probable inhibition of SOC and/or TRP channels. To evaluate the involvement of TRP channels in the vasorelaxant effect induced by carvacrol, non-selective inhibitors were used. No change in the relaxation response was observed in the presence of ruthenium red (pD2= 4.31  0.029, N=6), however, the effect induced by carvacrol was potentiated by La3+ (pD2 = 5.231  0,04, N=6), Gd3+ (pD2 = 4.97  0.02, N=6) or Ni2+ (pD2 = 5.079  0.02, N=6), furthermore, Mg2+ (pD2 = 4.168  0.021; MR = 81.12  4.03%, N=6) attenuated the relaxation elicited by carvacrol, suggesting that monoterpenoid may to action on TRPC1, TRPC3, TRPC6 and TRPM7 channels. Carvacrol also induced hypotension and bradycardia in non-anesthetized normotensive rats. In conclusion, these results suggest that carvacrol induced vasorelaxant effect in superior mesenteric artery rats isolated probably inhibiting Ca2+ influx by Cav, SOC (TRPC1), ROC (TRPC1 or TRPC6) and TRPM7 channels. Moreover, the effects induced by carvacrol in normotensive non-anesthetized rats showed a hypotensive and bradycardic activity. / Os efeitos farmacológicos de carvacrol, um fenol monoterpenóide, sobre o sistema cardiovascular, foi estudado em ratos normotensos, usando técnicas in vivo e in vitro. Carvacrol (10-8 - 3 ₓ 10-4 M) induziu vasorelaxamento dos anéis de artéria mesentérica superior isolada de rato pré-contraídos com 10 μM FEN (pD2 = 4,59  0,02, Emáx = 103,03  1,5%) na presença do endotélio funcional e esse efeito não foi alterado após a remoção do endotélio (pD2 = 4,36  0,02, Emáx = 111,03  4,8%), sugerindo, portanto, que a resposta vasorelaxante induzida por carvacrol parece ser independente do endotélio vascular. Interessantemente em anéis pré-contraídos com KCl 80 mM (pD2 = 4,12  0,01, Emáx = 94,38  3,97%), observou-se uma diminuição na sua potência e na sua eficácia farmacológica, sugerindo um passo comum na via que seria um aumento citosólico dos níveis de cálcio. Adicionalmente, carvacrol antagonizou, de maneira dependente de concentração, as contrações induzidas por CaCl2 em meio despolarizante nominalmente sem Ca2+ e induziu relaxamento das contrações induzidas pelo S(-)-Bay K 8644 (pD2 = 4,537  0,023, Emáx = 91,8  3,58%) com uma diminuição na sua eficácia farmacológica, sugerindo uma inibição do influxo de cálcio por canais de Ca2+ tipo-L. Além disso, antagonizou as contrações induzidas por CaCl2 em meio nominalmente sem cálcio, na presença de FEN e nifedipina, sugerindo uma provável inibição do influxo de cálcio por SOC, ROC e/ou canais TRP. Como também, em um meio despolarizante e nominalmente sem cálcio na presença de nifedipina esse mesmo antagonismo foi observado, ressaltando a provável inibição dos SOC e/ou canais TRP. Para avaliar a participação dos canais TRP, as preparações foram incubadas com La3+ (pD2 = 5,231  0,04) , Gd3+ (pD2 = 4,97  0,02) e Ni2+ (pD2 = 5,079  0,02) onde seu efeito foi potencializado sugerindo sua ação sobre os canais TRPC e ao utilizar magnésio (pD2 = 4,168  0,021 e Emáx = 81,12  4,03%) tanto sua potência quanto sua eficácia farmacológica foi atenuada, sugerindo inibição do canal TRPM7. Nos estudos in vivo, em ratos normotensos não anestesiados, carvacrol produziu hipotensão e bradicardia. Em conclusão, esses resultados sugerem que carvacrol induz efeito vasorelaxante em anéis de artéria mesentérica superior isolada de rato por inibir provavelmente TRPM7, como também inibir o influxo de cálcio por Cav, SOC, ROC e ou TRPC1 e 6. Além disso, os efeitos induzidos por carvacrol em ratos normotensos não anestesiados mostrou uma atividade hipotensora e bradicárdica.

This thesis deals with the determination of carvacrol, thymol, and eugenol by HPLC with electrochemical detection. Carbon paste electrode and boron doped diamond film electrode were used as the working electrodes. For the comparison, UV spectrophotometric detection at 275 nm was used besides the electrochemical detection. The separation was performed on LiChroCART 125-4, RP-18e (5 µm) column. Optimum separation conditions were found: mobile phase consisting of acetonitrile and acetate buffer in ratio 50:50, the optimum buffer pH was pH 5. The optimum potential of working electrode during electrochemical detection was +0,8 V and +1,2 V for carbon paste electrode and boron doped diamond film electrode, respectively. For both electrodes, repeatability of the measurement was examined; the surface of both electrodes had to be renewed between the measurements. Under the obtained optimum conditions, calibration dependences were measured. The studied substances were determined in real samples, in Thymus vulgaris L. and Achillea millefolium L. and in thyme-containing tea and syrup.