SILVA, A. K. O.; PESSOA, O. D. L. Estudo químico de espécies de Cordia (Boraginaceae): Cordia multispicata (Cham.) e Cordia globosa (Jacq.) 2013. 120 f. Dissertação (Mestrado em Química) - Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2013. / Submitted by José Jairo Viana de Sousa (jairo@ufc.br) on 2014-10-15T20:26:52Z No. of bitstreams: 1 2013_dis_akosilva.pdf: 5357295 bytes, checksum: 7cf7ea0232c8130d7dc69e9a9d4ecbd4 (MD5) / Approved for entry into archive by José Jairo Viana de Sousa(jairo@ufc.br) on 2015-11-24T20:05:34Z (GMT) No. of bitstreams: 1 2013_dis_akosilva.pdf: 5357295 bytes, checksum: 7cf7ea0232c8130d7dc69e9a9d4ecbd4 (MD5) / Made available in DSpace on 2015-11-24T20:05:34Z (GMT). No. of bitstreams: 1 2013_dis_akosilva.pdf: 5357295 bytes, checksum: 7cf7ea0232c8130d7dc69e9a9d4ecbd4 (MD5) Previous issue date: 2013 / This work reports the chemical investigation of the hexane extracts from the roots of Cordia multispicata and Cordia globosa species. For the compounds isolation were used classic chromatographic techniques, such as open and “flash” chromatography on column over silica gel, molecular exclusion on sephadex LH-20 and High Performance Liquid Cromatography in reverse phase. The chemical investigation of the hexane extract of C. multispicata lead to the isolation of two known terpenoid naphthoquinones, the cordiaquinones B and J. In addition, the new naphthoquinone 6-[10-(12,12,16-trimethyl-7- oxabicyclo[2.2.1]hept-2-yl]-2,3-dihydroxy-1,4-naphthalenedione, which was designed cordiaquinone P. From the hexane extract of C. globosa were isolated two uncommon terpenec hydroquinones, (4bE,6Z,8E)-1,4-dihydroxy-9a,10-dihydro-10,12-epoxy-5- methylbenzo[a]azulen-12-one and 4bZ,6Z,8E)-1-hydroxy-9a,10-dihydro-4,11:10,12- diepoxy-benzo[a]-azulen-11,12-dione, both reported for the first time. The Nbenzoylphenylalaninyl-N-benzoyl-2-amino-3-phenylpropyl was also isolated. The cytotoxic potential of the new compounds were evaluated against three tumor cell lines OVCAR-8 (ovarium), SF-295 (glioblastoma) and HCT-116 (colon). The compounds were weakly active showing IC50 values > 5µg/mL. The structural elucidation was performed by spectrometric methods: IR, HRMS and 1H and 13C NMR, including bidimensional techniques (COSY, HSQC and HMBC), in addition to comparison with literature data, whenever available. / O trabalho relata a investigação química dos extratos das raízes de Cordia multispicata e Cordia globosa. Para o isolamento dos compostos foram utilizadas técnicas cromatográficas clássicas, tais como: cromatografia em coluna aberta e do tipo “flash”, cromatografia por exclusão molecular e cromatografia líquida de alta eficiência. A investigação química do extrato hexânico de C. multispicata resultou no isolamento de duas naftoquinonas terpênicas conhecidas, as cordiaquinonas B e J, além da naftoquinona inédita 6-[10-(12,12,16-trimetil-7-oxabiciclo[2.2.1]hept-2-ila]-2,3-dihidroxi-1,4-naftalenodiona, a qual foi denominada de cordiaquinona P. Do extrato etanólico das raízes de C. globosa foram isoladas duas hidroquinonas terpênicas incomuns: (4bE,6Z,8E)-1,4-dihidroxi-9a,10-dihidro-10,12-epoxi-5-etilbenzo[a]azulen-12-ona e (4bZ,6Z,8E)-1-hidroxi-9a,10-dihidro-4,11:10,12-diepoxi-benzo[a]-azulen-11,12-diona, ambas relatadas pela primeira vez. O éster N-benzoil-L-fenilalaninato de N-benzoil-2-amino-3-fenilpropila foi também isolado. A citotoxicidade dos novos compostos foi avaliada frente a três linhagens de células tumorais: OVCAR-8 (ovário), SF-295 (glioblastoma) e HCT-116 (cólon). Os compostos foram fracamente ativos com valores de IC50 > 5 μg/mL. A determinação estrutural das substâncias isoladas foi realizada através de métodos espectrométricos: IR, EMAR and RMN 1H and 13C, incluindo técnicas bidimensionais (COSY, HSQC, HMBC), além de comparação com dados disponíveis na literatura, sempre que disponíveis.

Written records about medicinal plants date back at least 5,000 years to the Sumerians. The objected plants for present investigation were indigenous to South Africa and as explored, only a few biological studies were found on the previous studies on Hyaenanche globosa and Maytenus procumbens. Phytochemical studies of the ethanol extract of the fruits of H. g/obosa (F.E) resulted in isolation of two known pure sesquiterpene lactones; 'tutin 1' and 'hyenanchin 2'. The crude extract and its isolated constituents were tested on four cancerous and a normal cell lines. F.E exhibited the highest antiproliferative activity on Hela cells which followed by Caco-2 cells. None of the isolated compounds were found to be toxic to the cells tested in this experiment. F.E demonstrated potent inhibition of DPPH radical activity similar to vitamin C. 'Tutin 1' and 'hyenanchin 2' were found with marginal antioxidant activity of which 'compound 1' presented more potent activity than 'compound 2'. The amounts of ROS radicals formed by pure compounds (1 and 2) were not significantly higher than those of controls. This is the first report on phytochemical index, anticancer, antioxidant and antibacterial properties of F.E and its purified compounds. The possible biochemical activities of the acetonic/ethanolic extract of the leaves of Maytenus procumbens (L.M.P), and its isolated compounds were investigated in the present study. L.M.P showed IC50 values of 68.79, 51.22, 78.49, 76.59 and 76.64 ì/ml on Caco-2, Hela, HT29, NIH3T3 and T47D cells by use of MTT cytotoxicity assay. Bioassay guided fractionation led to the isolation and identification of two new triterpenes: '30-hydroxy-11á-hydroxy-18â-olean-12-en-3-one 3' and '30-hydroxy-11á-methoxy- 18â-olean-12-en-3-one 5'. In addition, a known terpenoid: 'asiatic acid 4' was purified. Due to the unavailability of sufficient amount of 'asiatic acid 4', this compound was not tested. Pure compounds 3 and 5 exhibited the most cytotoxicity against Hela cells and were further investigated for their abilities for induction of apoptosis (at the concentration of their IC sub>50) in Hela cells using flow cytometric method. Both compounds induced apoptosis up to 73.20%, (compound 3) and 20.40% (compound 5) in Hela cells versus control group (0.40%). Antioxidant/oxidative properties of L.M.P and its isolated compounds were investigated using extracellular (DPPH), and intracellular reactive oxygen species (ROS) assays. L.M.P and the isolated compounds exhibited marginal DPPH discoloration. Experimental samples represented a time and concentrationdependent function of ROS formation in Hela cells. ROS generation might be a part of the mechanisms by which compounds 3 and 5 induced apoptosis in Hela cells. It can therefore be concluded that the active components in L.M.P might serve as a mediator of the reactive oxygen scavenging system and have the potential to act as a prooxidant and an antioxidant, depending on the biological environment of the cells. There is no report until date on phytochemical index, anticancer, antioxidant and antibacterial properties of L.M.P and its isolated compounds. / Thesis (PhD)--University of Pretoria, 2012. / Plant Science / unrestricted

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / O trabalho relata a investigaÃÃo quÃmica dos extratos das raÃzes de Cordia multispicata e Cordia globosa. Para o isolamento dos compostos foram utilizadas tÃcnicas cromatogrÃficas clÃssicas, tais como: cromatografia em coluna aberta e do tipo âflashâ, cromatografia por exclusÃo molecular e cromatografia lÃquida de alta eficiÃncia. A investigaÃÃo quÃmica do extrato hexÃnico de C. multispicata resultou no isolamento de duas naftoquinonas terpÃnicas conhecidas, as cordiaquinonas B e J, alÃm da naftoquinona inÃdita 6-[10-(12,12,16-trimetil-7-oxabiciclo[2.2.1]hept-2-ila]-2,3-dihidroxi-1,4-naftalenodiona, a qual foi denominada de cordiaquinona P. Do extrato etanÃlico das raÃzes de C. globosa foram isoladas duas hidroquinonas terpÃnicas incomuns: (4bE,6Z,8E)-1,4-dihidroxi-9a,10-dihidro-10,12-epoxi-5-etilbenzo[a]azulen-12-ona e (4bZ,6Z,8E)-1-hidroxi-9a,10-dihidro-4,11:10,12-diepoxi-benzo[a]-azulen-11,12-diona, ambas relatadas pela primeira vez. O Ãster N-benzoil-L-fenilalaninato de N-benzoil-2-amino-3-fenilpropila foi tambÃm isolado. A citotoxicidade dos novos compostos foi avaliada frente a trÃs linhagens de cÃlulas tumorais: OVCAR-8 (ovÃrio), SF-295 (glioblastoma) e HCT-116 (cÃlon). Os compostos foram fracamente ativos com valores de IC50 > 5 μg/mL. A determinaÃÃo estrutural das substÃncias isoladas foi realizada atravÃs de mÃtodos espectromÃtricos: IR, EMAR and RMN 1H and 13C, incluindo tÃcnicas bidimensionais (COSY, HSQC, HMBC), alÃm de comparaÃÃo com dados disponÃveis na literatura, sempre que disponÃveis. / This work reports the chemical investigation of the hexane extracts from the roots of Cordia multispicata and Cordia globosa species. For the compounds isolation were used classic chromatographic techniques, such as open and âflashâ chromatography on column over silica gel, molecular exclusion on sephadex LH-20 and High Performance Liquid Cromatography in reverse phase. The chemical investigation of the hexane extract of C. multispicata lead to the isolation of two known terpenoid naphthoquinones, the cordiaquinones B and J. In addition, the new naphthoquinone 6-[10-(12,12,16-trimethyl-7- oxabicyclo[2.2.1]hept-2-yl]-2,3-dihydroxy-1,4-naphthalenedione, which was designed cordiaquinone P. From the hexane extract of C. globosa were isolated two uncommon terpenec hydroquinones, (4bE,6Z,8E)-1,4-dihydroxy-9a,10-dihydro-10,12-epoxy-5- methylbenzo[a]azulen-12-one and 4bZ,6Z,8E)-1-hydroxy-9a,10-dihydro-4,11:10,12- diepoxy-benzo[a]-azulen-11,12-dione, both reported for the first time. The Nbenzoylphenylalaninyl-N-benzoyl-2-amino-3-phenylpropyl was also isolated. The cytotoxic potential of the new compounds were evaluated against three tumor cell lines OVCAR-8 (ovarium), SF-295 (glioblastoma) and HCT-116 (colon). The compounds were weakly active showing IC50 values > 5Âg/mL. The structural elucidation was performed by spectrometric methods: IR, HRMS and 1H and 13C NMR, including bidimensional techniques (COSY, HSQC and HMBC), in addition to comparison with literature data, whenever available.

Submitted by Bruna Rodrigues (bruna92rodrigues@yahoo.com.br) on 2017-10-04T13:31:39Z No. of bitstreams: 1 DissRKR.pdf: 1036463 bytes, checksum: 2c845de1e471e5e42f4a7e8591961fc9 (MD5) / Approved for entry into archive by Ronildo Prado (producaointelectual.bco@ufscar.br) on 2017-10-10T20:14:17Z (GMT) No. of bitstreams: 1 DissRKR.pdf: 1036463 bytes, checksum: 2c845de1e471e5e42f4a7e8591961fc9 (MD5) / Approved for entry into archive by Ronildo Prado (producaointelectual.bco@ufscar.br) on 2017-10-10T20:14:25Z (GMT) No. of bitstreams: 1 DissRKR.pdf: 1036463 bytes, checksum: 2c845de1e471e5e42f4a7e8591961fc9 (MD5) / Made available in DSpace on 2017-10-10T20:21:54Z (GMT). No. of bitstreams: 1 DissRKR.pdf: 1036463 bytes, checksum: 2c845de1e471e5e42f4a7e8591961fc9 (MD5) Previous issue date: 2017-05-02 / Não recebi financiamento / Soil is an environment with great microbial diversity. Filamentous fungi and bacteria have been known for decades for the ability to stimulate plant development. Recently, studies have shown that yeasts are also capable of benefiting plants, for example, through the solubilization of inorganic phosphates. This work aims at a better understanding of the ability of rhizospheric yeasts in the solubilization of tricalcium phosphate. It also aims to study the influence of factors (pH, temperature and agitation) in the process. Initially, isolates of corn and sugarcane rhizosphere yeasts were made. The isolates were evaluated for the ability to solubilize tricalcium phosphate in solid medium BDYA. The isolate with the highest solubilization capacity was identified and evaluated for the solubilization capacity of tricalcium phosphate in NBRIP liquid medium at different temperature levels (20, 25 and 30 ° C), initial pH (3, 5 and 7) and agitation (0, 100, 150 and 200 rpm). Solubilization evaluations were also performed at 24-hour intervals, ranging from 1 to 7 days. Six isolates were obtained. All presented a positive result for solubilization. The yeast 6S01 (Torulaspora globosa) had the highest solubilization capacity. After five days, T. globosa solubilized 26 to 35% of the total phosphorus in the NBRIP culture medium. There was no significant difference for any of the environmental parameters tested. Assays performed in periods of 1 to 7 days showed that the yeast solubilized a maximum amount of phosphorus in 48 hours (42%). This makes it interesting for industrial and agricultural applications, since it has a fast solubilization process and can be used in a wide range of conditions. It was possible to understand that a higher cell growth is not related to the efficiency in the solubilization, and that acidification of the medium may be one of the main mechanisms used by yeast in the in vitro solubilization of tricalcium phosphate. / O solo é um ambiente com grande diversidade microbiana. Fungos filamentosos e bactérias são conhecidos há décadas pela capacidade de estimular o desenvolvimento vegetal. Recentemente, trabalhos mostram que leveduras também são capazes de beneficiar plantas, por exemplo, através da solubilização de fosfatos inorgânicos. Esse trabalho visa um maior entendimento da capacidade de leveduras rizosféricas na solubilização de fosfato tricálcico. Também visa o estudo da influência de fatores (pH, temperatura e agitação) no processo. Inicialmente, foi feito um isolamento de leveduras de rizosfera de milho e cana-de-açúcar. Os isolados foram avaliados quanto à capacidade de solubilizar fosfato tricálcico em meio sólido BDYA. O isolado com maior capacidade de solubilização foi identificado e avaliado quanto à capacidade de solubilização de fosfato tricálcico em meio líquido NBRIP, em diferentes níveis de temperatura (20, 25 e 30°C), pH inicial (3, 5 e 7) e agitação (0, 100, 150 e 200 rpm). Também foram feitas avaliações da solubilização em intervalos de 24 horas, de 1 a 7 dias. Foram obtidos seis isolados. Todos apresentaram resultado positivo para solubilização. A levedura 6S01 (Torulaspora globosa) apresentou a maior capacidade de solubilização. Após cinco dias, T. globosa solubilizou 26 a 35% do fósforo total no meio de cultura NBRIP. Não houve diferença significativa para nenhum dos parâmetros ambientais testados. Ensaios realizados em períodos de 1 a 7 dias mostraram que a levedura solubilizou uma quantidade máxima de fósforo em 48 horas (42%). Isso a torna interessante para aplicações industriais e agrícolas, visto que possui rapidez no processo de solubilização e pode ser empregada numa ampla faixa de condições. Foi possível compreender que um maior crescimento celular não está relacionado à eficiência na solubilização, e que a acidificação do meio pode ser um dos principais mecanismos utilizados pela levedura na solubilização in vitro do fosfato tricálcico.

Submitted by Aelson Maciera (aelsoncm@terra.com.br) on 2017-03-30T19:00:59Z No. of bitstreams: 1 DissTBO.pdf: 1573981 bytes, checksum: e4e97b3faf18cb39cb38dffdf87ccfc6 (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2017-04-17T19:48:58Z (GMT) No. of bitstreams: 1 DissTBO.pdf: 1573981 bytes, checksum: e4e97b3faf18cb39cb38dffdf87ccfc6 (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2017-04-17T19:49:10Z (GMT) No. of bitstreams: 1 DissTBO.pdf: 1573981 bytes, checksum: e4e97b3faf18cb39cb38dffdf87ccfc6 (MD5) / Made available in DSpace on 2017-04-17T20:04:48Z (GMT). No. of bitstreams: 1 DissTBO.pdf: 1573981 bytes, checksum: e4e97b3faf18cb39cb38dffdf87ccfc6 (MD5) Previous issue date: 2016-08-26 / Não recebi financiamento / Microorganisms Plant Growth Promoters (MPCV) have the ability to produce compounds which stimulate plant growth by different mechanisms, such as the production of plant hormones and solubilization of minerals. Considering these aspects, this study aimed to evaluate the potential of yeast strains as plant growth promoters by producing indole acetic acid (IAA) and phosphate solubilization, and its inoculation of tomato seedlings. Six yeast strains were evaluated belonging to bank microorganisms Agricultural Microbiology and Molecular Laboratory (LAMAM - CCA, UFSCar). For the evaluation of IAA production by isolates the colorimetric method was used, modified DiMenna (1957); for quantification of tricalcium phosphate solubilization was used to colorimetric method of molybdenum blue. The test with tomato seedlings was conducted in a greenhouse with different cell concentrations of T. globosa (5S55) and with or without addition of glucose and tryptophan. The results showed that isolates of T. globosa and Rh. mucilaginosa produced IAA in the presence of tryptophan; the isolates of T. globosa and M. guilliermondii solubilized inorganic phosphate in a liquid medium. The isolate T. globosa (5S55) promoted the growth of tomato seedlings, with significant increases in the length of shoot and root. In conclusion, the isolates of T. globosa, Rh. mucilaginosa and M. guilliermondii have the potential for improving plant production by IAA production and / or phosphate solubilization; the yeast T. globosa (5S55) was shown to promote growth of tomato plants. / Micro-organismos Promotores de Crescimento Vegetal (MPCV) apresentam a habilidade de produzir compostos que estimulam o desenvolvimento das plantas por meio de diferentes mecanismos, como a produção de hormônios vegetais e a solubilização de minerais. Considerando estes aspectos, este trabalho teve por objetivo avaliar o potencial de linhagens de leveduras como promotoras de crescimento vegetal, através da avaliação in vitro da produção de ácido indolacético e solubilização de fosfato, e da avaliação in vivo através da inoculação da linhagem com melhores resultados em mudas de tomate. Foram avaliadas seis linhagens de levedura pertencentes ao banco de micro-organismos do Laboratório de Microbiologia Agrícola e Molecular (LAMAM - CCA, UFSCar). Para a avaliação da produção de AIA pelos isolados foi utilizada a metodologia colorimétrica, modificada de diMenna (1957); para quantificação da solubilização de fosfato tricálcico foi utilizada o método colorimétrico do azul de molibdênio. O ensaio com mudas de tomate foi realizado em casa de vegetação com diferentes concentrações de células de T. globosa (5S55), e com adição ou não de glicose e triptofano. Os resultados mostraram que os isolados de T. globosa e o isolado de Rhodotorula mucilaginosa produziram AIA na presença de triptofano; os isolados de T. globosa e o isolado de Meyerozyma guilliermondii solubilizaram fosfato inorgânico em meio líquido. A levedura T. globosa (5S55) promoveu o crescimento das mudas de tomate, com aumentos significativos no comprimento da parte aérea e raiz. Conclui-se que os isolados de T. globosa, Rh. mucilaginosa e M. guilliermondii apresentam potencial para a melhoria da produção vegetal através da produção de AIA e/ou solubilização de fosfato; a levedura T. globosa, isolado 5S55, mostrou ser capaz de promover o crescimento de mudas de tomate.

Ce travail porte sur la dynamique bactérienne (abondances totale, vivante et active, productions de cellules et de biomasse, activités de dégradation exo-enzymatique) en Manche orientale, depuis une approche écosystémique à l'application d'outils microbiens spécifiques. Il permet de mieux comprendre le rôle des bactéries, jusqu'alors peu étudiées en Manche orientale, dans les flux de matière et d'énergie, particulièrement en période de floraison printanière.<br />Le premier objectif consistait à déterminer la réponse du bactérioplancton à la variabilité spatio-temporelle du milieu, à l'échelle (i) de l'écosystème au cours de deux années consécutives (2003/2004), (ii) saisonnière sur le site de Wimereux et (iii) journalière au cours de suivis Lagrangiens de masse d'eau. Cette étude montre que les années 2003 et 2004 ont été très différentes en termes de fonctionnement trophique du réseau microbien. Par ailleurs, la diversité des paramètres de contrôle de la dynamique bactérienne (température, disponibilité en matières organiques, pression de prédation) conditionne son caractère saisonnier. Sur un cycle journalier, l'application de deux méthodes de mesure de la production a permis une analyse plus précise et plus fine des conditions de croissance bactérienne.<br />Afin d'évaluer le rôle trophique du bactérioplancton, particulièrement lors des blooms massifs de Phaeocystis globosa, les fractions cellulaires viables et actives du pool bactérien, responsables de l'activité mesurée, ont été estimées. Les abondances de ces cellules augmentent significativement en réponse aux matières organiques accumulées pendant et après le bloom phytoplanctonique. Des mesures d'activité exo-enzymatique montrent une dégradation bactérienne significative des matières issues du bloom, et soulignent l'existence d'une étroite relation « trophique » entre bactéries et P. globosa.

Background: Polyphenols are secondary metabolites produced by plants to defend themselves from environmental stressors. We explored the effect of Wolffia globosa ‘Mankai’, a novel cultivated strain of a polyphenol-rich aquatic plant, on the metabolomic-gut clinical axis in vitro, in-vivo and in a clinical trial. Methods: We used mass-spectrometry-based metabolomics methods from three laboratories to detect Mankai phenolic metabolites and examined predicted functional pathways in a Mankai artificial-gut bioreactor. Plasma and urine polyphenols were assessed among the 294 DIRECT-PLUS 18-month trial participants, comparing the effect of a polyphenol-rich green-Mediterranean diet (+1240 mg/polyphenols/day, provided by Mankai, green tea and walnuts) to a walnuts-enriched (+440 mg/polyphenols/day) Mediterranean diet and a healthy controlled diet. Results: Approximately 200 different phenolic compounds were specifically detected in the Mankai plant. The Mankai-supplemented bioreactor artificial gut displayed a significantly higher relative-abundance of 16S-rRNA bacterial gene sequences encoding for enzymes involved in phenolic compound degradation. In humans, several Mankai-related plasma and urine polyphenols were differentially elevated in the green Mediterranean group compared with the other groups (p < 0.05) after six and 18 months of intervention (e.g., urine hydroxy-phenyl-acetic-acid and urolithin-A; plasma Naringenin and 2,5-diOH-benzoic-acid). Specific polyphenols, such as urolithin-A and 4-ethylphenol, were directly involved with clinical weight-related changes. Conclusions: The Mankai new plant is rich in various unique potent polyphenols, potentially affecting the metabolomic-gut-clinical axis.

Background: Rare plants that contain corrinoid compounds mostly comprise cobalamin analogues, which may compete with cobalamin (vitamin B12 (B12)) metabolism. We examined the presence of B12 in a cultivated strain of an aquatic plant: Wolffia globosa (Mankai), and predicted functional pathways using gut-bioreactor, and the effects of long-term Mankai consumption as a partial meat substitute, on serum B12 concentrations. Methods: We used microbiological assay, liquid-chromatography/electrospray-ionization-tandem-mass-spectrometry (LC-MS/MS), and anoxic bioreactors for the B12 experiments. We explored the effect of a green Mediterranean/low-meat diet, containing 100 g of frozen Mankai shake/day, on serum B12 levels during the 18-month DIRECT-PLUS (ID:NCT03020186) weight-loss trial, compared with control and Mediterranean diet groups. Results: The B12 content of Mankai was consistent at different seasons (p = 0.76). Several cobalamin congeners (Hydroxocobalamin(OH-B12); 5-deoxyadenosylcobalamin(Ado-B12); methylcobalamin(Me-B12); cyanocobalamin(CN-B12)) were identified in Mankai extracts, whereas no pseudo B12 was detected. A higher abundance of 16S-rRNA gene amplicon sequences associated with a genome containing a KEGG ortholog involved in microbial B12 metabolism were observed, compared with control bioreactors that lacked Mankai. Following the DIRECT-PLUS intervention (n = 294 participants; retention-rate = 89%; baseline B12 = 420.5 ± 187.8 pg/mL), serum B12 increased by 5.2% in control, 9.9% in Mediterranean, and 15.4% in Mankai-containing green Mediterranean/low-meat diets (p = 0.025 between extreme groups). Conclusions: Mankai plant contains bioactive B12 compounds and could serve as a B12 plant-based food source.

O complexo Sporothrix schenckii reúne espécies etiologicamente relacionadas à esporotricose, uma micose que pode acometer seres humanos e animais. Foi realizada a identificação fenotípica e molecular de 85 isolados provenientes de quatro estados brasileiros (Minas Gerais, Rio de Janeiro, Rio Grande do Sul e São Paulo). Foram pesquisadas a produção de enzimas, o perfil de inibição por leveduras killer, a suscetibilidade aos antifúngicos comerciais e aos extratos de plantas. Os isolados foram identificados como S. schenckii, S. brasiliensis e S. globosa, com predomínio de S. schenckii. Houve discordância de 37,7% entre a identificação das espécies do complexo S. schenckii utilizando metodologias fenotípicas e genotípicas. Entre os antifúngicos testados, a terbinafina foi o fármaco mais ativo; seguido por cetoconazol e itraconazol, enquanto fluconazol e voriconazol foram os menos ativos. Cinco isolados fúngicos foram detectados como resistentes ao itraconazol, sendo um S. globosa e quatro S. schenckii. Não houve diferença nos perfis de suscetibilidade aos antifúngicos entre as espécies do complexo Sporothrix schenckii. Entre os extratos de origem vegetal, o mais ativo foi o proveniente de Pterocaulon polystachyum, mostrando que o uso popular das plantas reforça a importância de pesquisas etnofarmacológicas, abrindo a possibilidade de encontrar novos agentes antifúngicos clinicamente eficazes. Doze das 18 leveduras killer avaliadas apresentaram atividade frente a todos os isolados do complexo S. schenckii estudados. No entanto, não houve diferença na suscetibilidade as toxinas entre as espécies de Sporothrix. Todos os isolados produziram desoxiribonuclease, urease e proteinase. Atividade fosfolipase e esterase foi detectada em 83 (97,6%) e 80 (94,1%), respectivamente, dos isolados testados. Todas as amostras do complexo S. schenckii produziram, pelo menos, quatro das enzimas avaliadas, e 78 (91,8%) dos isolados produziram todas as enzimas analisadas no estudo. No entanto, não foi possível diferenciar as espécies de Sporothrix baseado no perfil enzimático. Entre as enzimas extracelulares avaliadas nos isolados do complexo S. schenckii, desoxiribonuclease e esterase foram produzidas em maior quantidade, podendo vir a ser um fator de virulência. Além disso, o caldo Sabouraud dextrose mostrou potencial para ser usado na avaliação in vitro da atividade antifúngica frente ao complexo S. schenckii. / The Sporothrix schenckii complex combines species etiologically related to sporotrichosis, a mycosis which can affect humans and animals. The phenotypic and genotypic identification of 85 strains from four Brazilian States (Minas Gerais, Rio de Janeiro, Rio Grande do Sul and São Paulo) was performed. The enzymatic production, profile of inhibition by killer yeasts, susceptibility to marketed antifungal and to plant extracts were surveyed. The isolates were identified as S. schenckii, S. brasiliensis and S. globosa, with the predominance of S. schenckii. There was 37.7% disagreement regarding the species classification using phenotypic and genotypic methodologies. Among the tested antifungals, terbinafine was the most active drug, followed by ketoconazole and itraconazole, while fluconazole and voriconazole were the least active ones. Five isolates - one S. globosa and four S. schenckii - were resistent to itraconazole. There was no difference as to the profiles of the susceptibility to the antifungal agents among the Sporothrix species. The most active vegetal extract was from Pterocaulon polystachyum, showing that the popular use of these plants reinforces the importance of ethnopharmacological researches, with the possibility of finding new clinically effective antifungal agents. Twelve out of the 18 evaluated killer yeasts showed activity against all the tested strains of the S. schenckii complex. However, there was no difference in susceptibility to the toxins among the Sporothrix species complex. All the isolates were desoxiribonuclease, urease and proteinase positive. Phospholipase and esterase activities were detected in 83 (97.6%) and 80 (94.1%), respectively, among the isolates evaluated. All the S. schenckii complex strains produced at least four of the evaluated enzymes, and 78 (91.8%) of the isolates produced all the enzymes analyzed in the study. However, it is not possible to differentiate the Sporothrix species based on their enzymatic profile. Among the extracellular enzymes evaluated in the S. schenckii complex isolates, desoxiribonuclease and esterase were the most prominent ones, and their production may be a virulence factor. Furthermore, the Sabouraud dextrose broth showed potential to be used in the in vitro evaluation of the antifungal activity against the S. schenckii complex.

O complexo Sporothrix schenckii reúne espécies etiologicamente relacionadas à esporotricose, uma micose que pode acometer seres humanos e animais. Foi realizada a identificação fenotípica e molecular de 85 isolados provenientes de quatro estados brasileiros (Minas Gerais, Rio de Janeiro, Rio Grande do Sul e São Paulo). Foram pesquisadas a produção de enzimas, o perfil de inibição por leveduras killer, a suscetibilidade aos antifúngicos comerciais e aos extratos de plantas. Os isolados foram identificados como S. schenckii, S. brasiliensis e S. globosa, com predomínio de S. schenckii. Houve discordância de 37,7% entre a identificação das espécies do complexo S. schenckii utilizando metodologias fenotípicas e genotípicas. Entre os antifúngicos testados, a terbinafina foi o fármaco mais ativo; seguido por cetoconazol e itraconazol, enquanto fluconazol e voriconazol foram os menos ativos. Cinco isolados fúngicos foram detectados como resistentes ao itraconazol, sendo um S. globosa e quatro S. schenckii. Não houve diferença nos perfis de suscetibilidade aos antifúngicos entre as espécies do complexo Sporothrix schenckii. Entre os extratos de origem vegetal, o mais ativo foi o proveniente de Pterocaulon polystachyum, mostrando que o uso popular das plantas reforça a importância de pesquisas etnofarmacológicas, abrindo a possibilidade de encontrar novos agentes antifúngicos clinicamente eficazes. Doze das 18 leveduras killer avaliadas apresentaram atividade frente a todos os isolados do complexo S. schenckii estudados. No entanto, não houve diferença na suscetibilidade as toxinas entre as espécies de Sporothrix. Todos os isolados produziram desoxiribonuclease, urease e proteinase. Atividade fosfolipase e esterase foi detectada em 83 (97,6%) e 80 (94,1%), respectivamente, dos isolados testados. Todas as amostras do complexo S. schenckii produziram, pelo menos, quatro das enzimas avaliadas, e 78 (91,8%) dos isolados produziram todas as enzimas analisadas no estudo. No entanto, não foi possível diferenciar as espécies de Sporothrix baseado no perfil enzimático. Entre as enzimas extracelulares avaliadas nos isolados do complexo S. schenckii, desoxiribonuclease e esterase foram produzidas em maior quantidade, podendo vir a ser um fator de virulência. Além disso, o caldo Sabouraud dextrose mostrou potencial para ser usado na avaliação in vitro da atividade antifúngica frente ao complexo S. schenckii. / The Sporothrix schenckii complex combines species etiologically related to sporotrichosis, a mycosis which can affect humans and animals. The phenotypic and genotypic identification of 85 strains from four Brazilian States (Minas Gerais, Rio de Janeiro, Rio Grande do Sul and São Paulo) was performed. The enzymatic production, profile of inhibition by killer yeasts, susceptibility to marketed antifungal and to plant extracts were surveyed. The isolates were identified as S. schenckii, S. brasiliensis and S. globosa, with the predominance of S. schenckii. There was 37.7% disagreement regarding the species classification using phenotypic and genotypic methodologies. Among the tested antifungals, terbinafine was the most active drug, followed by ketoconazole and itraconazole, while fluconazole and voriconazole were the least active ones. Five isolates - one S. globosa and four S. schenckii - were resistent to itraconazole. There was no difference as to the profiles of the susceptibility to the antifungal agents among the Sporothrix species. The most active vegetal extract was from Pterocaulon polystachyum, showing that the popular use of these plants reinforces the importance of ethnopharmacological researches, with the possibility of finding new clinically effective antifungal agents. Twelve out of the 18 evaluated killer yeasts showed activity against all the tested strains of the S. schenckii complex. However, there was no difference in susceptibility to the toxins among the Sporothrix species complex. All the isolates were desoxiribonuclease, urease and proteinase positive. Phospholipase and esterase activities were detected in 83 (97.6%) and 80 (94.1%), respectively, among the isolates evaluated. All the S. schenckii complex strains produced at least four of the evaluated enzymes, and 78 (91.8%) of the isolates produced all the enzymes analyzed in the study. However, it is not possible to differentiate the Sporothrix species based on their enzymatic profile. Among the extracellular enzymes evaluated in the S. schenckii complex isolates, desoxiribonuclease and esterase were the most prominent ones, and their production may be a virulence factor. Furthermore, the Sabouraud dextrose broth showed potential to be used in the in vitro evaluation of the antifungal activity against the S. schenckii complex.