Phenylalanine 4-monooxygenase and the S-oxidation of S-carboxymethyl-L-cysteine by human cytosolic fractions.

The purpose of this investigation was to reaction phenotype the identity of the cytosolic enzyme responsible for the S-oxidation of S-carboxymethyl-L-cysteine (SCMC) in female human hepatic cytosolic fractions. The identity of this enzyme in the female Wistar rat hepatic cytosolic fraction was found to be phenylalanine 4-monooxygenase (PAH). In pooled female human hepatic cytosolic fractions the calculated K(m) and V(max) for substrate (SCMC) activated PAH was 16.22 +/- 11.31 mM and 0.87 +/- 0.41 nmoles x min(-1) mg(-1). The experimental data modelled to the Michaelis-Menten equation with noncompetitive substrate inhibition. When the cytosolic fractions were activated with lysophophatidylcholine the V(max) increased to 52.31 +/- 11.72 nmoles x min(-1) mg(-1) but the K(m) remained unchanged at 16.53 +/- 2.32 mM. A linear correlation was seen in the production of Tyr and SCMC R/S S-oxide in 20 individual female hepatic cytosolic fractions for both substrate and lysophosphatidylcholine activated PAH (r(s) > 0.96). Inhibitor studies found that the specific chemical and antibody inhibitors of PAH reduced the production of Tyr and SCMC R/S S-oxide in these in vitro PAH assays. An investigation of the mechanism of interaction of SCMC with PAH indicated that the drug was a competitive inhibitor of the aromatic C-oxidation of Phe with a calculated K(i) of 17.23 +/- 4.15 mM. The requirement of BH4 as cofactor and the lack of effect of the specific tyrosine hydroxylase, tryptophan hydroxylase and nitric oxide synthase inhibitors on the S-oxidation of SCMC all indicate that PAH was the enzyme responsible for this biotransformation reaction in human hepatic cytosolic fractions

Phenylalanine 4-monooxygenase and the S-oxidation of S-carboxymethyl-L-cysteine by human cytosolic fractions

The purpose of this investigation was to reaction phenotype the identity of the cytosolic enzyme responsible for the S-oxidation of S-carboxymethyl-L-cysteine (SCMC) in female human hepatic cytosolic fractions. The identity of this enzyme in the female Wistar rat hepatic cytosolic fraction was found to be phenylalanine 4-monooxygenase (PAH). In pooled female human hepatic cytosolic fractions the calculated K(m) and V(max) for substrate (SCMC) activated PAH was 16.22 +/- 11.31 mM and 0.87 +/- 0.41 nmoles x min(-1) mg(-1). The experimental data modelled to the Michaelis-Menten equation with noncompetitive substrate inhibition. When the cytosolic fractions were activated with lysophophatidylcholine the V(max) increased to 52.31 +/- 11.72 nmoles x min(-1) mg(-1) but the K(m) remained unchanged at 16.53 +/- 2.32 mM. A linear correlation was seen in the production of Tyr and SCMC R/S S-oxide in 20 individual female hepatic cytosolic fractions for both substrate and lysophosphatidylcholine activated PAH (r(s) > 0.96). Inhibitor studies found that the specific chemical and antibody inhibitors of PAH reduced the production of Tyr and SCMC R/S S-oxide in these in vitro PAH assays. An investigation of the mechanism of interaction of SCMC with PAH indicated that the drug was a competitive inhibitor of the aromatic C-oxidation of Phe with a calculated K(i) of 17.23 +/- 4.15 mM. The requirement of BH4 as cofactor and the lack of effect of the specific tyrosine hydroxylase, tryptophan hydroxylase and nitric oxide synthase inhibitors on the S-oxidation of SCMC all indicate that PAH was the enzyme responsible for this biotransformation reaction in human hepatic cytosolic fractions

Morphology and ecology of the planktonic diatom Palmerina hardmaniana (Greville) Hasle in southern Brazil

The diatom Palmerina hardmaniana (Greville) Hasle presents a wide geographical distribution in neritic tropical and subtropical regions. In the present work we analyzed plankton samples collected monthly between 1995 and 2007 at the surf zone of Cassino Beach, RS (32° 12’ S and 52° 10’ W), and in winter 2005 and summer 2007 at the continental shelf and slope in southern Brazil, Santa Marta Grande Cape, SC and Albardão-Chuí, RS regions (28° 23’-33° 07’ S and 48° 41’-52° 26’ W). We present the detailed morphological description of P. hardmaniana, and the first study including electron scanning microscope observations for material from the southwestern Atlantic Ocean. The morphometric data confirm the identity of the species in all its ultra-structural details. Palmerina hardmaniana was only observed in summer-autumn months with low cell density (< 500 cells.L–1) at both Cassino Beach surf zone and coastal shelf stations. The warm water temperature (18-29 °C) indicates the most probable origin of its inoculum are tropical/subtropical regions. Salinities of 23-36 and the relatively high silicate content indicate the importance of the terrestrial discharge during occasions when P. hardmaniana was observed, probably with influence on the nutrient availability. We emphasize that the species was not cited previously for Argentinean and Uruguayan waters and suggest that the southern Brazilian region is close to the southern geographical distribution limit of Palmerina hardmaniana in the southwestern Atlantic Ocean.A diatomácea Palmerina hardmaniana(Greville) Hasle apresenta ampla distribuição geográfica em águas neríticas tropicais e subtropicais. No presente trabalho foram analisadas amostras de plâncton, coletadas mensalmente entre 1995 e 2007 na zona de rebentação da Praia do Cassino, RS (32° 12’ S e 52° 10’ W), e no inverno de 2005 e verão de 2007 na plataforma continental e talude do sul do Brasil, na região de Cabo de Santa Marta Grande, SC e Albardão-Chuí, RS (28° 23’-33° 07’ S e 48° 41’-52° 26’ W). Apresentamos a descrição detalhada de P. hardmaniana, como primeiro estudo com observações de microscopia eletrônica de varredura para material coletado em águas do Oceano Atlântico Sul Ocidental. Os dados morfológicos analisados confirmam a identificação da espécie em todos os seus detalhes estruturais. Palmerina hardmaniana somente foi observada nos meses de verão/outono, em baixa densidade (< 500 células.L–1) na zona de arrebentação da Praia do Cassino bem como em estações costeiras da plataforma continental. A temperatura quente da água (18-29 °C), indica as águas tropicais/subtropicais como possível origem do inóculo de P. hardmaniana no verão-outono. A salinidade entre 23 e 36 e o teor relativamente alto de sílica também indicam a importância da descarga terrestre nas ocasiões de presença de P. hardmaniana, exercendo importante papel no suprimento de nutrientes. Salienta-se que a espécie não é citada em águas argentinas e uruguaias e assim, sugerimos que o extremo sul do Brasil representa aproximadamente o limite sul da distribuição geográfica de Palmerina hardmaniana no Oceano Atlântico Sul Ocidental