Caracterização química, atividade antioxidante e antimicrobiana do óleo essencial de Baccharis oreophila Malme

The Baccharis oreophila Malme belongs to the Asteraceae family. In Brazil are reported 120 species of Baccharis, most located in the South and Southeast regions, the latter presents the highest prevalence, especially in the state of São Paulo. Asteraceae is well known for the production of essential oils, which are liquid, volatile and aromatic substances produced by plants specialized for metabolism possess antibacterial, antifungal, and antioxidant properties. Thus, this study aimed, perform chemical and evaluate the antimicrobial and antioxidant activity of essential oil from dried leaves of B. oreophila collected in winter in Piraquara, Paraná. Obtaining essential oil was given by hydrodistillation in Clevenger apparatus, in triplicate, and the analysis was done using a gas chromatograph coupled to mass spectrometry GC / MS. The identification of the components was made based on retention indices calculated from the co-injection of a series of n-alkanes, followed by comparison of their mass spectra with literature. The antimicrobial activity was assessed by disk diffusion method and microdilution. The antioxidant activity was evaluated by the methods DPPH equivalent Trolox, ABTS and FRAP equivalent Trolox equivalent ferrous sulfate. The essential oil showed 0.47% yield. They identified 57 components (89.38%), 1.51% were classified as hydrogenated monoterpenes, oxygenated monoterpenes 15.14%, 34.84% and 37.87% hydrogenated sesquiterpenes sesquiterpenes oxygenates. As the major components were detected kusimono (16.37%), spathulenol (16.12%), the δ-cadinene (5.68%) and bicyclogermacrene (4.09%). The antimicrobial activity of essential oil was performed for the microorganisms Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Candida albicans ATCC 18804 and Candida tropicalis ATCC 13803, the results showed that the essential oil showed activity against S. aureus Inhibitory Concentration minimum (CIM) 1250 g/mL. In the evaluation of antioxidant activity essential oil showed antioxidant potential for the three methods evaluated, with values of 1,468 m.mol.L-1, 7.126 m.mol.L-1 and 45.515 m.mol.L-1 for ABTS, DPPH and FRAP, respectively. These results demonstrate that the essential oil of B. oreophila showed antimicrobial potential against S. aureus and interesting antioxidant activity, especially for the reducing power of iron ion, demonstrating their potential for future industrial applications. It is important to emphasize that were not observed in the literature reports highlighting such biological properties of B. oreophila oil.

Bioactivity And Chemical Composition Of The Essential Oil From The Leaves Of Guatteria Australis A.st.-hil.

Essential oil from the leaves of Guatteria australis was obtained by hydrodistillation, analyzed by Gas Chromatography coupled to Mass Spectromery (GC-MS) and their antiproliferative, antileishmanial, antibacterial, antifungal and antioxidant activities were also evaluated. Twenty-three compounds were identified among which germacrene B (50.66%), germacrene D (22.22%) and (E)-caryophyllene (8.99%) were the main compounds. The highest antiproliferative activity was observed against NCI-ADR/RES (TGI = 31.08 μg/ml) and HT-29 (TGI = 32.81 μg/ml) cell lines. It also showed good antileishmanial activity against Leishmania infantum (IC50 = 30.71 μg/ml). On the other hand, the oil exhibited a small effect against Staphylococcus aureus ATCC 6538, S. aureus ATCC 14458 and Escherichia coli ATCC 10799 (MIC = 250 μg/ml), as well as small antioxidant activity (457 μmol TE/g) assessed through ORACFL assay. These results represent the first report regarding chemical composition and bioactivity of G. australis essential oil.

Eletroforese capilar acoplada à espectrometria de massas (CE-MS): vinte anos de desenvolvimento

CE-MS has been increasingly used for analysis of a vast array of compounds. This article reviews the different electrophoretic modes, interfaces and mass analyzers that are commonly used in the CE-MS coupling, as well as the technique advantages and performance characteristics. A large compilation of CE-MS applications is also presented. Therefore, this review is both a guide for beginners and a collection of key references for people who are familiar to the technique. Furthermore, this is the first CE-MS review published in a Brazilian journal and marks the installation of the first two commercial CE-MS units in Sao Paulo State.

New analytical methodologies for doping control – detection of anabolic androgenic steroids in human urine

Dissertation submitted to Faculdade de Ciências e Tecnologia - Universidade Nova de Lisboa in fulfilment of the requirements for the degree of Doctor of Philosophy (Biochemistry - Biotechnology)

Larvicidal activity of the essential oil from Lippia sidoides cham. against Aedes aegypti linn

The aim of this work was to study the larvicidal activity of Lippia sidoides essential oil against Aedes aegypti larvae. The essential oil and its hydrolate (saturated solution of essential oil in water) were obtained by vapor extraction and their chemical composition determined by GL-chromatography coupled to mass spectroscopy. Bioassays were run with the essential oil, pure and diluted hydrolate and with their main constituents thymol and carvacrol. The results obtained showed that L. sidoides essential oil and its hydrolate have larvicidal action against the mosquito A. aegypti, causing an almost instantaneous mortality. Thymol, an alkylated phenol derivative and one of the major components of L. sidoides essential oil, was identified as the active principle responsible for the larvicidal action, causing 100% larval mortality at the lowest tested concentration of 0.017% (w/v). These results suggest that the essential oil of L. sidoides is promising as larvicide against A. aegypti and could be useful in the search of newer, more selective, and biodegradable larvicidal natural compounds to be used in official combat programs and at home.

Larvicidal Activity of essential oils from Brazilian plants against Aedes aegypti L.

Aedes aegypti L. is the major vector of dengue fever, an endemic disease in Brazil. In an effort to find effective and affordable ways to control this mosquito, the larvicidal activities of essential oils from nine plants widely found in the Northeast of Brazil were analyzed by measurement of their LC50. The essential oils were extracted by steam distillation and their chemical composition determined by GL-chromatography coupled to mass spectroscopy. The essential oils from Cymbopogon citratus and Lippia sidoides, reported in the literature to have larvicidal properties against A. aegypti, were used for activity comparison. The results show that Ocimum americanum and Ocimum gratissimum have LC50 of 67 ppm and 60 ppm respectively, compared to 63 ppm for L. sidoides and 69 ppm for C. citratus. These results suggest a potential utilization of the essential oil of these two Ocimum species for the control of A. aegypti.

FM127: Systemic metal ion levels in patients with modular neck total hip arthroplasties: a prospective cohort study

There is increasing evidence that modular neck stems are prone to corrosion-related complications. Recent studies showed elevated metal ions levels and occasional pseudotumor formation in patients with such implants. The purpose of this study was to compare systemic metal-ion levels in patients after primary THA with modular neck stems to those of patients after non-modular implants. To our knowledge, this is the first cohort study including a control group, THA without CoCr heads and dry-assembled neck-stem connections. Methods: 50 patients after THA at a minimum follow-up of 1 year have been selected for the study. Patients with multiple prosthesis or other implants have been deselected. All received a cementless SPS stem from Symbios (Ti6Al4V). 40 patients have the modular neck (CoCr) version and 10 a monobloc version. All bearings were either ceramic-ceramic or ceramic-polyethylene to minimize other sources of CoCr ion release. In the modular group, the neck was chosen pre-operatively based on a 3D planning, allowing for a dry assembly of the stem and neck on the back table before implantation. A plasma system coupled to mass spectrometry was used for a complete elementary quantification in blood and serum separately. Clinical outcome was measured using the Oxford Hip Score. Results : Complete data sets of 29 patients (24 in the modular neck-group (10male, mean age 63y, 35-84y) and 5 in the monobloc-group (3 male, 69 y, 51-83y) are available to date. Mean Co blood levels were .95 ug/L (.14-12.4) in the modular group vs .27 ug/L (.10-.73) in the monobloc group (p=.2). Respective values for Cr were significantly higher in the modular group (.99 g/L; range .75-1.21) compared to those in the monobloc group (.74 g/L ;.62-.86; p=.001). No significant difference was found when comparing serum levels. 5/24 patients had Co levels above 1 g/L (12/24 for Cr), which is by some considered as a relevant elevation. The maximum Co level was measured in an asymptomatic patient. The Oxford Hip Scores were similar in both groups. Conclusion: Cr levels were significantly elevated in the modular neck group compared to those in the monobloc group. 1/24 patients with a modular prosthesis exhibited Co levels, which are beyond the threshold accepted even for metal-on-metal bearing couples. These results have contributed to our decision to abandon the use of modular neck stems. Routine follow-up including annual measurements of systemic CoCr concentrations should be considered.

Caracterização cromatográfica de compostos orgânicos presentes nos resíduos sólidos provenientes de indústria de reciclagem de papel e sua aplicação na produção de briquetes de carvão vegetal

Recycling of paper in industrial scale has become an established practice worldwide. In this work, organic compositions of three different kinds of sludge generated in recycle paper industry were studied, and the incorporation of one of those sludge in briket was also investigated. The characterization of organic compounds in sludge samples and briket was performed using Gas Chromatography coupled with Mass Spectrometry after a Soxhlet extraction. Different chemical classes were identified in each type of sludge, but just the sludge composed by cellulose residue did not presented polyaromatic hydrocarbons. Four formulations of sludge incorporated with charcoal for briket production were evaluated.

PYROLIGNEOUS LIQUOR PRODUCED FROM ACACIA MEARNSII DE WILD WOOD UNDER CONTROLLED CONDITIONS AS A RENEWABLE SOURCE OF CHEMICALS

Acacia mearnsii de Wild (black wattle) is one of the most important trees planted in Southern Brazil for tannin extraction and charcoal production. The pyrolysis of the black wattle wood used for obtaining charcoal is performed in brick ovens, with the gas fraction being sent directly into the environment. The present study examines the condensable compounds present in the liquor produced from black wattle wood at different thermal degradation conditions, using gas chromatography coupled with mass spectrometry (GC/MS). Branches of black wattle were thermally degraded at controlled ambient and temperature conditions. Overall, a higher variety of compounds were obtained under atmospheric air pressure than under synthetic air pressure. Most of the tentatively identified compounds, such as carboxylic acids, phenols, aldehydes, and low molecular mass lignin fragments, such as guayacol, syringol, and eugenol, were products of lignin thermoconversion. Substituted aromatic compounds, such as vanillin, ethyl vanillin, and 2-methoxy-4-propeny-phenol, were also identified. At temperatures above 200 ºC, furan, 2-acetylfuran, methyl-2-furoate, and furfural, amongst others, were identified as polysaccharide derivatives from cellulose and hemicellulose depolymerization. This study evidences the need for adequate management of the condensable by-products of charcoal production, both for economic reasons and for controlling their potential environmental impact.

Antimicrobial activity of extracellular metabolites from antagonistic bacteria isolated from potato (Solanum phureja) crops

Microorganisms for biological control are capable of producing active compounds that inhibit the development of phytopathogens, constituting a promising tool toob tain active principles that could replace synthetic pesticides. This study evaluatedtheability of severalpotentialbiocontrol microorganismsto produce active extracellular metabolites. In vitro antagonistic capability of 50 bacterial isolates from rhizospheric soils of "criolla" potato (Solanum phureja) was tested through dual culture in this plant with different plant pathogenic fungi and bacteria. Isolates that showed significantly higher antagonistic activity were fermented in liquid media and crude extracts from the supernatants had their biological activities assessed by optical density techniques. Inhibitory effecton tested pathogens was observed for concentrations between 0.5% and 1% of crude extracts. There was a correlation between the antimicrobial activity of extracts and the use of nutrient-rich media in bacteria fermentation. Using a bioguided method, a peptidic compound, active against Fusarium oxysporum, was obtained from the 7ANT04 strain (Pyrobaculum sp.). Analysis by nuclear magnetic resonance and liquid chromatography coupled to mass detector evidenced an 11-amino acid compound. Bioinformatic software using raw mass data confirmed the presence of a cyclic peptide conformed by 11 mostly non-standard amino acids.

Cartographie du réseau d'interactions protéiques de la machinerie de transcription dans les cellules humaines

Les protéines sont les macromolécules les plus polyvalentes de la cellule. Elles jouent un rôle fondamental dans la majorité des processus biologiques à travers la formation de complexes multi-protéiques. Durant la transcription, une multitude de facteurs sont impliquées dans le contrôle de l’activité des complexes ARN polymérases. Notre laboratoire s’est intéressé au réseau d’interaction de la machinerie de transcription des ARN polymérases nucléaires, dans le but de mieux comprendre leurs mécanismes de régulation. Pour ce faire, une procédure protéomique comprenant la purification de complexes protéiques par affinité couplée à la spectrométrie de masse et à l’analyse bioinformatique a été développée. La méthode de purification TAP (Tandem Affinity Purification) a été adaptée pour permettre la purification de complexes protéiques solubles assemblés in vivo à partir de cellules humaines. L’objectif de mon projet de maîtrise était de purifier le complexe de l’ARN Pol I ainsi que de poursuivre l’expansion du réseau d’interactions protéine-protéine de la machinerie de transcription de l’ARN Pol II humaine. À l’aide des protéines POLR1E, TWISTNB, POLR2E, PFDN4, MBD2, XPA, CAND1 et PDCD5 étiquetées (TAP-tag) exprimées dans des lignées cellulaires ECR-293, plusieurs complexes protéiques solubles ont été purifiés et analysés par spectrométrie de masse. Les interactions protéiques ont été triées et validées bioinformatiquement pour donner en final une liste d’interactions ayant un haut degré de confiance à partir de laquelle des réseaux d’interactions protéine-protéine ont été créés. Le réseau créé au cours de ce projet connecte plusieurs composantes de la machinerie transcriptionnelle tels que les ARN Pol I, II et III, les complexes RPAP3/R2TP/prefoldin-like, TRiC/CCT, Mi-2/NuRD et des facteurs de transcription et de réparation de l’ADN. Ce type d’analyse nous a permis d’identifier et de caractériser de nouveaux régulateurs de la machinerie de transcription de l’ARN Pol I et II et de mieux comprendre son fonctionnement.

Découverte de nouvelles interactions entre le virus de l'Hépatite C et l'hôte par une approche combinée de Spectrométrie de Masse et de Génomique Fonctionnelle

La réplication et l’assemblage du virus de l’hépatite C (VHC) sont régulés finement dans le temps et l’espace par les interactions protéiques entre le virus avec l’hôte. La compréhension de la biologie du virus ainsi que sa pathogénicité passe par les connaissances relatives aux interactions virus/hôte. Afin d’identifier ces interactions, nous avons exploité une approche d’immunoprécipitation (IP) couplée à une détection par spectrométrie de masse (MS), pour ensuite évaluer le rôle des protéines identifiées dans le cycle viral par une technique de silençage génique. Les protéines virales Core, NS2, NS3/4A, NS4B, NS5A et NS5B ont été exprimées individuellement dans les cellules humaines 293T et immunoprécipitées afin d’isoler des complexes protéiques qui ont été soumis à l’analyse MS. Ainsi, 98 protéines de l’hôte ont été identifiées avec un enrichissement significatif et illustrant une spécificité d’interaction. L’enrichissement de protéines connues dans la littérature a démontré la force de l’approche, ainsi que la validation de 6 nouvelles interactions virus/hôte. Enfin, le rôle de ces interactants sur la réplication virale a été évalué dans un criblage génomique par ARN interférant (ARNi). Deux systèmes rapporteurs de la réplication virale ont été utilisés : le système de réplicon sous-génomique (Huh7-Con1-Fluc) et le système infectieux (J6/JFH-1/p7Rluc2a), ainsi qu’un essai de toxicité cellulaire (Alamar Blue). Parmi les protéines de l’hôte interagissant avec le VHC, 28 protéines ont démontré un effet significatif sans effet de toxicité cellulaire, suggérant fortement un rôle dans la réplication du VHC. Globalement, l’étude a mené à l’identification de nouvelles interactions virus/hôte et l’identification de nouvelles cibles thérapeutiques potentielles.

Multifunctional acid formation from the gas-phase ozonolysis of beta-pinene

The gas-phase ozonolysis of beta-pinene was studied in static chamber experiments, using gas chromatography coupled to mass spectrometric and flame ionisation detection to separate and detect products. A range of multifunctional organic acids-including pinic acid, norpinic acid, pinalic-3- acid, pinalic-4-acid, norpinalic acid and OH-pinalic acid-were identified in the condensed phase after derivatisation. Formation yields for these products under systematically varying reaction conditions (by adding different OH radical scavengers and Criegee intermediate scavengers) were investigated and compared with those observed from alpha-pinene ozonolysis, allowing detailed information on product formation mechanisms to be elucidated. In addition, branching ratios for the initial steps of the reaction were inferred from quantitative measurements of primary carbonyl formation. Atmospheric implications of this work are discussed.

Formation of organic acids from the gas-phase ozonolysis of terpinolene

Gas-phase ozonolysis of terpinolene was studied in static chamber experiments using gas chromatography coupled to mass spectrometric and flame ionisation detection to separate and detect products. Two isomers of C-7-diacids and three isomers of C-7-aldehydic acids were identified in the condensed phase after derivatisation. Possible mechanisms of formation of these acids were investigated using different OH radical scavengers and relative humidities, and were compared to those reported earlier for the ozonolysis of beta-pinene. In addition, branching ratios for some of the individual reaction steps, e. g. the branching ratio between the two hydroperoxide channels of the C-7-CI, were deduced from the quantitative product yield data. Branching ratios for POZ decomposition and the stabilisation/decomposition of the C-7-CI were also obtained from measurements of the C-7 primary carbonyl product.

Produção de fitomassa, teor e produtividade do óleo essencial de Baccharis dracunculifolia DC. em função da adubação orgânica

Objetivou-se avaliar a produção de fitomassa, teor e produtividade do óleo essencial de Baccharis dracuculifolia DC., em função de doses de adubo orgânico, e foram utilizadas mudas produzidas por semente, e mantidas sob sombrite pelo período de 84 dias até serem transplantadas para o campo, no espaçamento de 1,0 x 1,0 m, com irrigação por gotejamento. O delineamento estatístico utilizado foi em blocos ao acaso, com quatro repetições, e as doses de composto orgânico foram: 0, 10, 20, 30, 40 e 50 t ha-1. Foi realizada uma colheita, aos 150 dias após o transplante das mudas. O óleo essencial foi extraído por hidrodestilação e analisado em cromatógrafo a gás acoplado a espectrômetro de massas (Shimadzu, QP-5000). Todos os resultados foram submetidos à análise de variância pelo teste F seguido da análise de regressão e o teste Tukey para os dados: massa seca útil da parte aérea, teor e produtividade do óleo essencial. Para as variáveis massa seca total e massa seca útil da parte aérea, as dosagens estudadas influenciaram as plantas, que apresentaram os melhores resultados na dose 50 t ha-1. Para teor de óleo não houve influência significativa para as dosagens estudadas, porém a dose 30 t ha-1 apresentou o melhor resultado. Entretanto para a produtividade do óleo essencial, as dosagens estudadas influenciaram as plantas, que na dosagem 30 t ha-1 obtiveram os melhores resultados.