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In vitro activity of doripenem and comparator antimicrobial agents was evaluated against Gram-negative bacilli recently isolated from Brazilian private hospitals that were enrolled in the INVITA-A-DORI Brazilian Study. A total of 805 unique Gram-negative bacilli were collected from patients hospitalized at 18 medical centers between May/08 and March/09. Each hospital was asked to submit 50 single Gram-negative bacilli isolated from blood, lower respiratory tract or intraabdominal secretions. Bacterial identification was confirmed and antimicrobial susceptibility testing was performed using Clinical Laboratory Standards Institute (CLSI) microdilution method at a central laboratory. CLSI M100-S21 (2011) or US-FDA package insert criteria (tigecycline) was used for interpretation of the antimicrobial susceptibility results. Doripenem was as active as meropenem and more active than imipenem against E. coli and K. pneumoniae isolates. A total of 50.0% of Enterobacter spp. isolates were resistant to ceftazidime but 85.7% of them were inhibited at doripenem MICs ≤ 1 μg/mL. Polymyxin B was the only agent to show potent activity against Acinetobacter spp. (MIC50/90, ≤ 0.5/1 μg/mL) and P. aeruginosa (MIC50/90, 1/2 μg/mL). Although high rates of imipenem (53.1%) and meropenem (44.5%) resistance were detected among P. aeruginosa, doripenem showed MIC50 of 16 μg/mL against imipenem-resistant P. aeruginosa and inhibited a greater number of imipenem-resistant P. aeruginosa (10.5%) at MIC values of ≤ 4 μg/mL than did meropenem (0.0%). In this study, doripenem showed similar in vitro activity to that of meropenem and retained some activity against imipenem-resistant P. aeruginosa isolated from Brazilian medical centers. Keywords: carbapenems, Gram-negative bacteria, Brazil

In vitro activity of doripenem and comparator antimicrobial agents was evaluated against Gram-negative bacilli recently isolated from Brazilian private hospitals that were enrolled in the INVITA-A-DORI Brazilian Study. A total of 805 unique Gram-negative bacilli were collected from patients hospitalized at 18 medical centers between May/08 and March/09. Each hospital was asked to submit 50 single Gram-negative bacilli isolated from blood, lower respiratory tract or intraabdominal secretions. Bacterial identification was confirmed and antimicrobial susceptibility testing was performed using Clinical Laboratory Standards Institute (CLSI) microdilution method at a central laboratory. CLSI M100-S21 (2011) or US-FDA package insert criteria (tigecycline) was used for interpretation of the antimicrobial susceptibility results. Doripenem was as active as meropenem and more active than imipenem against E. coli and K. pneumoniae isolates. A total of 50.0% of Enterobacter spp. isolates were resistant to ceftazidime but 85.7% of them were inhibited at doripenem MICs < 1 µg/mL. Polymyxin B was the only agent to show potent activity against Acinetobacter spp. (MIC50/90, < 0.5/1 µg/mL) and P. aeruginosa (MIC50/90, 1/2 µg/mL). Although high rates of imipenem (53.1%) and meropenem (44.5%) resistance were detected among P. aeruginosa, doripenem showed MIC50 of 16 µg/mL against imipenem-resistant P. aeruginosa and inhibited a greater number of imipenem-resistant P. aeruginosa (10.5%) at MIC values of < 4 µg/mL than did meropenem (0.0%). In this study, doripenem showed similar in vitro activity to that of meropenem and retained some activity against imipenem-resistant P. aeruginosa isolated from Brazilian medical centers.

Strawberry (Fragaria x ananassa Duch.) fruits contain phenolic compounds that have antioxidant, anticancer, antiatherosclerotic and anti-neurodegenerative properties. Identification of food phenolics is necessary since their nature, size, solubility, degree and position of glycosylation and conjugation influence their absorption, distribution, metabolism and excretion in humans. Freeze-dried whole strawberry fruit powder and strawberry fruit extracts were analyzed by liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) methods. Phenolics were identified as ellagic acid (EA), EA-glycosides, ellagitannins, gallotannins, anthocyanins, flavonols, flavanols and coumaroyl glycosides. The anthocyanidins were pelargonidin and cyanidin, found predominantly as their glucosides and rutinosides. The major flavonol aglycons were quercetin and kaempferol found as their glucuronides and glucosides. LC-ESI-MS/MS methods differentiated EA from quercetin conjugates since both aglycons have identical molecular weights (302 g/mol). The identification of strawberry phenolics is necessary to generate standardized materials for in vitro and in vivo studies and for the authentication of strawberry-based food products. (c) 2005 Elsevier Ltd. All rights reserved.

Pomegranate (Punica granatum L.) fruits are widely consumed fresh and in the forms of juice, concentrate, wine and jam. Pomegranate fruit husk is a rich source of hydrolyzable tannins called ellagitannins (ETs). During processing methods in the commercial pomegranate juice (PJ) industry, ETs are extracted from the fruit husk in significant quantities into the juice. Pomegranate husk, a by-product of the PJindustry, is therefore an inexpensive and abundant source of ETs which are present in PJ. Previous methods to isolate pomegranate ETs included labor intensive and time-consuming solid phase extractions by column chromatography (C-18, polyamides, cellulose, Sephadex Lipophilic LH-20, Diaion HP20) and/or use of specialized instruments such as preparative-high performance liquid chromatography (HPLC). We have used an Amberlite XAD-16 resin vacuum-aspirated column to rapidly purify an aqueous extract of pomegranate husk to afford total pomegranate tannins (TPT) in substantial yields (58-60 g TPT/Kg husk; time

Background: Ellagic acid (EA) and hydrolyzable ellagitannins (ETs) are dietary polyphenols found in fruits and nuts and implicated with potent antioxidant, anticancer and antiatherosclerotic biological properties. Unfortunately, there are no reports on bioavailability studies of EA or ETs in the human body. Therefore we conducted in vivo studies whereby a human subject consumed pomegranate juice (180 ml) containing EA (25 mg) and ETs (318 mg, as punicalagins, the major fruit ellagitannin). Methods: A rapid plasma extraction procedure utilizing acidic precipitation of proteins, followed by HPLC-UV analyses was employed. Results: EA was detected in human plasma at a maximum concentration (31.9 ng/ml) after 1h post ingestion but was rapidly eliminated by 4h. The calibration curve for quantification of EA was linear (r2 = 0.9975) over the concentration range from 1000 to 15.6 ng/ml. Conclusions: Since EA has reportedly strong affinity for proteins and poor absorption in small animals, further studies to investigate whether the presence of free EA in human plasma may be due to its release from the hydrolysis of ETs, facilitated by physiological pH and/or gut microflora action, is warranted. EA can be considered as a biomarker for future human bioavailability studies involving consumption of ETs from food sources.

Pomegranate (Punica granatum L.) fruits are widely consumed as juice (PJ). The potent antioxidant and anti-atherosclerotic activities of PJ are attributed to its polyphenols including punicalagin, the major fruit ellagitannin, and ellagic acid (EA). Punicalagin, EA, a standardized total pomegranate tannin extract (TPT) and PJ were evaluated for in vitro antioxidant, antiproliferative and apoptotic activities. The antioxidative bioassays used included inhibition of lipid peroxidation and Trolox Equivalent Antioxidant Capacity (TEAC) assays. The antiproliferative assays targeted human oral (KB, CAL27), colon (HT-29, HCT116, SW480, SW620) and prostate (RWPE-1, 22Rv1) tumor cells. Apoptotic effects were evaluated against the HT-29 and HCT116 colon cancer cell lines. Punicalagin, EA and TPT were evaluated at 12.5-100 g/mL for antiproliferative assays. However, to evaluate the synergistic and/or additive contributions from other PJ phytochemicals, PJ was tested at concentrations normalized to deliver equivalent amounts of punicalagin (w/w). Punicalagin, EA, TPT and PJ were all evaluated at 10 g/mL concentrations for antioxidant properties and at 100 g/mL concentrations for apoptotic effects. PJ showed greatest antiproliferative activity against all cell lines by inhibiting proliferation from 30-100%. At 100 g/mL, PJ, EA, punicalagin and TPT induced apoptosis in HT-29 colon cells. However, in the HCT116 colon cells, EA, punicalagin and TPT but not PJ induced apoptosis. The trend in antioxidant activity was PJ>TPT>punicalagin>EA. The superior bioactivity of PJ compared to its purified polyphenols illustrated the multifactorial effects and chemical synergy of the action of multiple compounds compared to single purified active ingredients.

Strawberry (Fragaria x ananassa Duch.) fruits contain phenolic compounds that have antioxidant, anticancer, antiatherosclerotic and anti-neurodegenerative properties. Identification of food phenolics is necessary since their nature, size, solubility, degree and position of glycosylation and conjugation influence their absorption, distribution, metabolism and excretion in humans. Freeze-dried whole strawberry fruit powder and strawberry fruit extracts were analyzed by liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) methods. Phenolics were identified as ellagic acid (EA), EA-glycosides, ellagitannins, gallotannins, anthocyanins, flavonols, flavanols and coumaroyl glycosides. The anthocyanidins were pelargonidin and cyanidin, found predominantly as their glucosides and rutinosides. The major flavonol aglycons were quercetin and kaempferol found as their glucuronides and glucosides. LC-ESI-MS/MS methods differentiated EA from quercetin conjugates since both aglycons have identical molecular weights (302 g/mol). The identification of strawberry phenolics is necessary to generate standardized materials for in vitro and in vivo studies and for the authentication of strawberry-based food products.

Cranberries (Vaccinium macrocarpon Ait.) are an excellent dietary source of phytochemicals that include flavonol glycosides, anthocyanins, proanthocyanidins (condensed tannins), and organic and phenolic acids. Using C-18 and Sephadex Lipophilic LH-20 column chromatography, HPLC and tandem LC-ES/MS, we have analyzed, quantified and separated total cranberry extract (TCE) into fractions enriched in sugars, organic acids, total polyphenols, proanthocyanidins and anthocyanins (39.4, 30.0, 10.6, 5.5, 1.2% composition, respectively). Using a luminescent ATP cell viability assay, the antiproliferative effects of TCE (200 g/mL) vs. all fractions were evaluated against human oral (KB, CAL27), colon (HT-29, HCT116, SW480, SW620) and prostate (RWPE-1, RWPE-2, 22Rv1) cancer cell lines. The total polyphenol fraction was the most active fraction against all cell lines with 96.1 and 95% inhibition of KB and CAL27 oral cancer cells, respectively. For the colon cancer cells, the antiproliferative activity of this fraction was greatest against HCT116 (92.1%) than HT-29 (61.1%), SW480 (60%) and SW620 (63%). TCE and all fractions, showed 50% antiproliferative activity against prostate cancer cells with total polyphenols being the most active fraction (RWPE-1, 95%; RWPE-2, 95%; 22Rv1, 99.6%). Cranberry sugars (78.8 g/mL) did not inhibit the proliferation of any cancer cell lines. The enhanced antiproliferative activity of total polyphenols compared to TCE and its individual phytochemicals suggests synergistic or additive antiproliferative interactions of the anthocyanins, proanthocyanidins and flavonol glycosides within the cranberry extract.

Background: Ellagic acid (EA) and hydrolyzable ellagitannins (ETs) are dietary polyphenols found in fruits and nuts and implicated with potent antioxidant, anticancer and antiatherosclerotic biological properties. Unfortunately, there are no reports on the bioavailability studies of EA or ETs in the human body. We conducted in vivo studies whereby a human subject consumed pomegranate juice (PJ) (180 ml) containing EA (25 mg) and ETs (318 mg, as punicalagins, the major fruit ellagitannin). Methods: A rapid plasma extraction procedure utilizing acidic precipitation of proteins, followed by HPLC-UV analyses, was employed. Results: EA was detected in human plasma at a maximum concentration (31.9 ng/ml) after 1 h post-ingestion but was rapidly eliminated by 4 h. The calibration curve for quantification of EA was linear (r(2) = 0.9975) over the concentration range from 1000 to 15.6 ng/ml. Conclusions: Since EA has reportedly strong affinity for proteins and poor absorption in small animals, further studies to investigate whether the presence of free EA in human plasma may be due to its release from the hydrolysis of ETs, facilitated by physiological pH and/or gut microflora action, is warranted. EA can be considered as a biomarker for future human bioavailability studies involving consumption of ETs from food sources. (C) 2004 Elsevier B.V. All rights reserved.

The current chemotherapeutic modalities for advanced colorectal cancer are limited. DNA-platinating drugs such as cisplatin have poor efficacy against this malignancy. The aim of this study was to identify genes that render survival advantage after cisplatin treatment in metastatic colon cancer. Cell lines SW480 (primary colon cancer) and SW620 (metastatic lesion from the same patient) were obtained from ATCC. Apoptosis was measured by FACS analysis of cisplatin-treated (0.01-10 mug/ml) and untreated cells. Simultaneous analysis of similar to1200 cDNAs was performed by microarray technique on untreated and treated cells from lines. Microarray results were confirmed by RT-PCR. The SW620 cell line was more resistant to apoptosis induced by cisplatin. Western blot analysis revealed equal expression of pro-caspases 3, 8, and 9 in both cell lines. Microarray analysis identified 15 genes and 9 expressed sequence tags (ESTs) significantly altered both by cell type (metastatic vs. non-metastatic) and treatment vs. non-treatment. Several of these transcripts are well-characterized genes including MCT, GAD67, P19, GSTM3, Cyclin D1, ATM, and CO-029 that have been implicated in various malignancies. In the present study, we have identified a set of genes responsible for apoptosis resistance following treatment with cisplatin in the late stages of carcinogenesis. Targeting these genes may increase chemotherapy effectiveness in advanced colon cancer and reduce toxicity in normal tissue.

RESUMO A presente pesquisa foi desenvolvida com o objetivo de avaliar o potencial do estipe de Euterpe edulis Martius, material vegetal desprezado por ocasião da colheita do palmito, como matéria-prima fibrosa alternativa para a produção de polpa celulósica kraft. Foram efetuadas análises física e química do material lignocelulósico, por espectroscopia no infravermelho e ressonância magnética nuclear. Determinou-se, também, a viscosidade intrínseca, o peso molecular e o grau de polimerização das polpas celulósicas depuradas, branqueadas e não branqueadas, de Euterpe edulis e de Eucalyptus urophylla, que foram comparados entre si. Os rendimentos em polpa celulósica bruta e depurada (59,60% e 54,10%, respectivamente) foram superiores ou equipararam-se àqueles obtidos pelo Eucalyptus urophylla (54,50% e 51,70%, respectivamente). A resistência a degradação química, da celulose obtidas das duas espécies, frente aos agentes deslignificadores foram equivalentes.

RESUMO Vários testes experimentais de propagação vegetativa foram realizados com a espécie Vernonia scorpiodes (Lam) Pers, entre os quais a reprodução assexuada nas modalidades estaquia e a alporquia proporcionaram os melhores resultados, atingindo, respectivamente 83% de eficiência de pegamento e 55% de enraizamento.

RESUMO: O método de Björkman de isolamento de lignina é utilizado em vários laboratórios de química da madeira do mundo inteiro. As evidências de modificações químicas da protolignina durante o processo de isolamento pelo método de Björkman tem sido alvo de interesse, já que, vários produtos de reação a partir de modelos dos tipos B-O-4 e B-5 são formados via reação de homólise em ambiente (moinho de bolas) gerador de energia mecânica.

RESUMO A lignificação é um processo bioquímico que abrange desde a formação dos monolignóis até a polimerização da lignina na parede celular. Além do apoio ao sistema estrutural da parede celular, defesa física e química, a lignina participa no processo de transporte de água no floema vascular e nas células do xilema. As estruturas moleculares das ligninas mostram-se variações complexas de acordo com sua composição. A biossíntese da lignina é composta de duas etapas. A primeira, consta da participação de uma série de enzimas como: PAL, C4H, 4CL, C3H, OMT, F5H, CCR e CAD, que catalisam a formação dos precursores intermediários e finais dentro do compartimento citoplasmático. Durante a formação desses precursores algumas dessas enzimas atuam sobre o núcleo aromático, introduzindo hidroxilas em C-3 e C-5 e as metilando em seguida, enquanto outras promovem a redução da cadeia lateral. Estudos demonstram que a regulação destas enzimas podem modificar a arquitetura molecular e o grau de polimerização da lignina. Atualmente técnicas apuradas de transgenia são usadas para regular as atividades enzimáticas durante a formação da lignina, resultando em diversos benefícios para o setor florestal, agropecuária e meio ambiente.

RESUMO A madeira é um produto originado de um metabolismo bioquímico complexo de uma árvore. Esse metabolismo tem sido parcialmente descrito com base em estudos químicos, bioquímicos, enzimáticos e genômicos. A lignificação, por exemplo, é um processo final de uma via metabólica expressamente associada à formação do lenho, ativando os mecanismos do desenvolvimento geral e principalmente da diferenciação de determinados tecidos. Esse processo de formação xilemática recai sobre um controle de fatores altamente diversificados como nutrição, doenças e sistema hormonal. Esta revisão apresenta os fitohormônios como agentes reguladores do processo de formação da madeira. Estes fitohormônios são substâncias especiais que controlam vários processos fisiológicos e morfológicos de uma planta, assumindo grande importância sobre o contexto anatômico, bioquímico, fisiológico e genético. Em alguns, casos o efeito sinergístico sobre a xilogênese controla o processo de lignificação, por exemplo, na regulação da atividade da peroxidase na parede celular.

RESUMO Esta pesquisa trata da lignificação em plantas cultivadas em condições de atmosfera controlada de oxigênio. A espécie utilizada foi a Ipomoea batatas (L.) Lam - batata doce, desenvolvida por micropropagação. A lignina foi extraída pelo método de Klason, sendo realizados posteriormente espectros infravermelhos, que revelaram um perfil estrutural diferente na lignina de plantas cultivadas em condições de atmosfera controlada de oxigênio, quando comparadas com as testemunhas. Plantas sob tratamento (0,1 kg/cm2 - O2) apresentaram aproximadamente 14,4 mg / 596,8 mg de amostra de tecido seco, enquanto as testemunhas apresentaram 5,3 mg. A quantidade de extrativos (extração em acetona) das plantas crescidas em atmosfera de oxigênio (0,1kg/cm2-O2)foi aproximadamente de 0,9495 mg do tecido da planta seca, enquanto as testemunhas apresentaram 0,9317 mg / 496,3 mg de amostras de tecido seco.

RESUMO As propriedades antifúngicas de 2 extratos, obtidos a partir de caules de Vernonia scorpioides (Lam.) Pers., foram testadas para Penicillium citrinum, produtor de micotoxina nefrotóxica. Os extratos foram utilizados em ensaio de antibios e pelo método de difusão em agar, nas concentrações de 1 mg. 3 mg e 5 mg para cada 100 l de diluente, originando halos de inibição com 40, 50 e 80 mm, respectivamente. Tal resultado sugeriu ação fungicida, uma vez que não se conseguiu, à partir dos tratamentos, realizar-se subcultivos em meio Czapeck-Dox isento de inibidores. Foi avaliada a toxidez desses extratos ativos para camundongos albinos, aos quais ministrou-se uma dose cem vezes superior à utilizada in vitro. Não houve interferência no estado geral, ou ganho de peso, dos animais tratados durante 6 dias subsequentes a administração oral dos extratos. Através dos espectros no infravermelho dos extratas ativos, observou-se a existência de sinais, característicos do estiramento de carbonilas lactônicas (com absorção entre 1750 cm-1 e 1770 cm-1).

RESUMO Os países de clima tropical detêm as maiores reservas de plantas potencialmente bioativas do Planeta. Dentre as inúmeras representantes que compõe o vasto arsenal vegetal que é a Mata Atlântica destaca-se a família Asteraceae pelo seu grande número de espécies e ampla distribuição nas regiões tropicais e subtropicais. A grande diversidade bioquímica e taxonomia complexa aliada a ampla utilização popular na forma de chás contra as mais diversas doenças tornam-a um escelente alvo de pesquisas de interesse farmacológico. Em várias espécies, diferentes atividades biológicas são atribuídas a principios ativos presentes em folhas e caules, atividade antitumoral, antiinflamatória, anti-histamínica e antifúngica, entre outras. O gênero Vernônia, em especial, com várias lactonas sesquiterpênicas já isoladas e com atividade biológica conhecida, é uma promissora fonte de novos compostos com atividade biológica de interesse agroindustrial a serem utilizados futuramente.

ABSTACT During million years (MY) plants made up an efficient mechanism for their owner protection, neutralizing oxygen radicals (superoxide) in the cell wall. Evaluation of the oxygen paticipation on plant secondary metabolism, especially associated with the lignin formation, suggested two oxidation mechanisms: vertical and horizontal oxidation. The latter is mediated by enzymatic mechanisms, contributing to the structural profile of modern plants. Vertical oxidation is carried out in the cell wall and is responsible by lignin accumulation, while horizontal oxidation is elaborated into the cytoplasmatic compartment. These mechanisms are independent and play an important strategy to prevent destructive oxygen effect on the plant cells.

Resumo O objetivo da pesquisa foi propor um modelo sistemático de identificação de madeiras através de análise metabolômica por Ressonância Magnética Nuclear do 13C (RMN 13C) constituindo um banco de dados de 13C. Neste trabalho foram utilizados os extratos obtidos com ciclohexano de madeiras do híbrido E. urograndis. Quatro espécimes com aproximadamente 15 anos de idades foram coletados em diferentes localidades. Os dados de RMN 13C mostraram que os espectros dos extratos ciclohexano da madeira do cerne (DAP) foram os mais representativos para o estudo do E. urograndis devido ao número de absorções verificadas. Com os dados foi possível traçar o perfil espectral comum para o híbrido do E. urograndis de cada região anatômica (alburno, zona de transição e cerne).

RESUMO Com o objetivo de se verificar uma possível relação entre: atividade biológica de Vernonia scorpioides, planta medicinal de potente atividade antifúngica, e níveis de alguns elementos nutricionais, foram realizadas dosagens de N-amino, N- total, potássio e fósforo em raízes, caules e folhas através de processos analíticos clássicos. Nas raízes, onde é obtido o extrato mais ativo, foram encontrados os menores teores de nutrientes, sugerindo uma relação inversamente proporcional a atividade antifúngica para ela descrita.