Resumo Diversas espécies do gênero Piper acumulam piperamidas como metabólitos secundários e, embora tenham relevante importância biológica, detalhes de suas vias biossintéticas ainda não foram descritos experimentalmente. Experimentos envolvendo reações enzimáticas e incorporação de precursores marcados foram realizados utilizando as espécies Piper tuberculatum e Piper arboreum. As atividades das enzimas fenilalanina amônia liase (PAL), que estão envolvidas na via geral dos fenilpropanoides, foram monitoradas pela conversão do aminoácido L-fenilalanina em ácido cinâmico. A atividade da enzima 4-hidroxilase (C4H) também foi observada em P. tuberculatum pela conversão do ácido cinâmico em ácido p-cumárico. L-[UL-14C]-fenilalanina foi administrada em folhas de P. tuberculatum e incorporada em piperina (1), 4,5-di-hidropiperina (2), fagaramida (4), trans-piplartina (7) e diidropiplartina (9). Em P. arboreum, foi incorporado apenas à piperamida 4,5-dihidropiperilina (3). L-[UL-14C]-lisina foi incorporada com sucesso em 4,5-diidropiperina (2), trans-piplartina (7) e cis-piplartina (8). Esses dados corroboram a proposta de origem biossintética mista de piperamidas com a porção aromática originada do ácido cinâmico (via do ácido chiquímico) e construção de amida chave com aminoácidos como precursores. biológica experimentalmente arboreum PAL, PAL , (PAL) fenilpropanoides Lfenilalanina L 4hidroxilase hidroxilase 4 C4H CH C H (C4H P pcumárico. pcumárico p cumárico. cumárico p-cumárico LUL14Cfenilalanina LULCfenilalanina UL 14C 1, 1 (1) 4,5dihidropiperina 45dihidropiperina dihidropiperina 4,5 di hidropiperina 5 2, 2 (2) 4, (4) transpiplartina trans piplartina 7 (7 9. 9 . (9) 4,5dihidropiperilina 45dihidropiperilina dihidropiperilina 3. 3 (3) LUL14Clisina LULClisina lisina 4,5diidropiperina 45diidropiperina diidropiperina cispiplartina cis 8. 8 (8) chiquímico (PAL LUL Cfenilalanina (1 5dihidropiperina 45 (2 (4 ( (9 5dihidropiperilina (3 Clisina 5diidropiperina (8

Abstract Several Piper species accumulate piperamides as secondary metabolites, and although they have relevant biological importance, many details of their biosynthetic pathways have not yet been described experimentally. Experiments involving enzymatic reactions and labeled precursor feeding were performed using the species Piper tuberculatum and Piper arboreum. The activities of the phenylalanine ammonia lyase (PAL) enzymes, which are involved in the general phenylpropanoid pathway, were monitored by the conversion of the amino acid L-phenylalanine to cinnamic acid. The activity of the 4-hydroxylase (C4H) enzyme was also observed in P. tuberculatum by converting cinnamic acid to p-coumaric acid. L-[UL-14C]-phenylalanine was fed into the leaves of P. tuberculatum and incorporated into piperine (1), 4,5-dihydropiperine (2), fagaramide (4), trans-piplartine (7), and dihydropiplartine (9). In P. arboreum, it was only incorporated into the piperamide 4,5-dihydropiperiline (3). L-[UL-14C]-lysine was successfully incorporated into the 4,5-dihydropiperine piperidine group (2), dihydropyridinone, and trans- (7) and cis-piplartine (8). These data corroborate the proposal of mixed biosynthetic origin of piperamides with the aromatic moiety originating from cinnamic acid (shikimic acid pathway) and key amide construction with amino acids as precursors. metabolites importance experimentally arboreum PAL (PAL enzymes pathway Lphenylalanine L 4hydroxylase hydroxylase 4 C4H CH C H (C4H P pcoumaric p coumaric LUL14Cphenylalanine LULCphenylalanine UL 14C 1, 1 , (1) 4,5dihydropiperine 45dihydropiperine dihydropiperine 4,5 5 2, 2 (2) 4, (4) transpiplartine trans piplartine 7, 7 9. 9 . (9) 4,5dihydropiperiline 45dihydropiperiline dihydropiperiline 3. 3 (3) LUL14Clysine LULClysine lysine dihydropyridinone (7 cispiplartine cis 8. 8 (8) shikimic precursors LUL Cphenylalanine (1 5dihydropiperine 45 (2 (4 (9 5dihydropiperiline (3 Clysine ( (8

Resumo Piper tuberculatum (Piperaceae) é uma espécie que acumula especialmente amidas como metabólitos secundários e diversas atividades biológicas dessa espécie foram relatadas anteriormente. No presente artigo, relatamos um estudo proteômico dessa espécie. Eletroforese bidimensional (2D SDS-PAGE) e espectrometria de massas (ESI-Q-TOF) foram utilizadas nesse estudos. Mais de cem spots e vários peptídeos foram identificados nesta espécie e as funções putativas desses peptídeos relacionadas a mecanismo de defesa como estresse biótico e abiótico foram atribuídos. As informações apresentadas ampliam a gama de informações moleculares dessa espécie.

Abstract Piper tuberculatum (Piperaceae) is a species that accumulates especially amides as secondary metabolites and several biological activities was previously reported. In this article, we report a proteomic study of P. tuberculatum. Bidimensional electrophoresis (2D SDS-PAGE) and mass spectrometry (ESI-Q-TOF) were used in this study. Over a hundred spots and various peptides were identified in this species and the putative functions of these peptides related to defense mechanism as biotic and abiotic stress were assigned. The information presented extend the range of molecular information of P. tuberculatum.

Resumo No nordeste do Brasil, as lagoas de estabilização são muito adequadas para o tratamento de águas residuárias por causa da disponibilidade relativamente grande de terra e das condições ambientais (por exemplo, altas temperaturas) favoráveis ao melhor desenvolvimento dos microorganismos. Entretanto, florações de cianobactérias potencialmente tóxicas podem afetar o uso dessas lagoas de tratamento, devido à consequente qualidade inferior do efluente. O objetivo deste estudo foi avaliar a dinâmica das comunidades de fitoplâncton e a ocorrência de cianobactérias em uma lagoa de maturação situada após duas lagoas em série. Temperatura, oxigênio dissolvido, pH, DBO, N e P foram medidos durante um período de quatro meses, durante o qual amostras foram coletadas na superfície e fundo em sete pontos de amostragem da lagoa. As comunidades de fitoplâncton das amostras coletadas foram também identificadas e classificadas utilizando-se um microscópio óptico convencional. Para avaliar os resultados utilizou-se a análise de variância e o teste de Tukey. Para as três divisões de fitoplâncton encontradas (Cyanophyta, Chlorophyta e Euglenophyta), não houve diferença significativa para as amostras de superfície e de fundo de um mesmo mês. Entretanto, ocorreu grande variação para as amostras dos diferentes meses; nos meses de abril e outubro houve uma predominância de Cyanophyta, ao passo que em setembro o predomínio na lagoa foi de Chlorophyta. Os fatores determinantes que favoreceram o predomínio de Cyanophyta foram a baixa carga orgânica superficial aplicada (entre 78 e 109 kg DBO.ha–1.d–1) e N:P ≤ 10. A presença de duas das espécies de Cyanophyta, Oscillatoria sp. e Microcystisaeruginosa, consideradas potencialmente tóxicas, indica que é necessária precaução quando se considera o destino final do efluente tratado e sugere a necessidade de avaliar os riscos e benefícios associados ao uso da tecnologia de tratamento.

Abstract In northeastern Brazil, stabilization ponds are very suitable for wastewater treatment because of the relative great land availability and environmental conditions (e.g., high temperature) favorable for microorganism optimal development. However, blooms of potentially toxic cyanobacteria may affect the use of these treatment ponds due to resulting effluent poor quality. The objective of this study was to evaluate the dynamics of phytoplankton communities and the occurrence of cyanobacteria in a maturation pond located immediately after a series of two ponds. Temperature, dissolved oxygen, pH, BOD, N, and P were measured during a period of four months when samples were collected from the surface and the bottom of 7 sampling points distributed inside the pond. The phytoplankton of collected samples was also identified and classified using a conventional optical microscopy. Analysis of variance and Tukey test were used to evaluate the results. The three phytoplankton divisions found (Cyanophyta, Chlorophyta, and Euglenophyta) did not change considerably through surface and bottom. However, they changed greatly over the sampled months; great dominance of Cyanophyta was found at April and October, while Chlorophyta dominated the lagoon in September. Low superficial organic loads (between 78 and 109 kg BOD.ha–1.d–1) and N:P ≤ 10 were the determinant factors that favored the predominance of Cyanophyta. The presence of two potentially toxic species of Cyanophyta, Oscillatoria sp. and Microcystis aeruginosa, indicates that caution is required when considering the final destination of treated effluent and suggests a need to assess the risks and benefits associated with the use of the treatment technology.

The distribution of polymorphisms related to glutathione S-transferases (GST) has been described in different populations, mainly for white individuals. We evaluated the distribution of GST mu (GSTM1) and theta (GSTT1) genotypes in 594 individuals, by multiplex PCR-based methods, using amplification of the exon 7 of CYP1A1 gene as an internal control. In São Paulo, 233 whites, 87 mulattos, and 137 blacks, all healthy blood-donor volunteers, were tested. In Bahia, where black and mulatto populations are more numerous, 137 subjects were evaluated. The frequency of the GSTM1 null genotype was significantly higher among whites (55.4%) than among mulattos (41.4%; P = 0.03) and blacks (32.8%; P < 0.0001) from São Paulo, or Bahian subjects in general (35.7%; P = 0.0003). There was no statistically different distribution among any non-white groups. The distribution of GSTT1 null genotype among groups did not differ significantly. The agreement between self-reported and interviewer classification of skin color in the Bahian group was low. The interviewer classification indicated a gradient of distribution of the GSTM1 null genotype from whites (55.6%) to light mulattos (40.4%), dark mulattos (32.0%) and blacks (28.6%). However, any information about race or ethnicity should be considered with caution regarding the bias introduced by different data collection techniques, specially in countries where racial admixture is intense, and ethnic definition boundaries are loose. Because homozygous deletions of GST gene might be associated with cancer risk, a better understanding of chemical metabolizing gene distribution can contribute to risk assessment of humans exposed to environmental carcinogens.