Novel 2-[(benzylamino)methyl]pyrrolidine-3,4-diol derivatives as alpha-mannosidase inhibitors and with antitumor activities against hematological and solid malignancies.

Novel alpha-mannosidase inhibitors of the type (2R,3R,4S)-2-({[(1R)-2-hydroxy-1-arylethyl]amino}methyl)pyrrolidine-3,4-diol have been prepared and assayed for their anticancer activities. Compound 30 with the aryl group=4-trifluoromethylbiphenyl inhibits the proliferation of primary cells and cell lines of different origins, irrespective of Bcl-2 expression levels, inducing a G2/Mcell cycle arrest and by modification of genes involved in cell cycle progression and survival.

Orosomucoid (alpha-1 acid glycoprotein) phenotyping by use of immobilized pH gradients with 8 M urea and immunoblotting. A new variant encountered in a population study.

Orosomucoid (ORM) phenotyping has been performed on 329 unrelated Swiss subjects, using immobilized pH gradients with 8 M urea and 2% v/v 2-mercaptoethanol followed by immunoblotting. After desialylation the band patterns of ORM confirmed that the polymorphism of the structural locus ORM1 is controlled by three codominant autosomal alleles (ORM1*F1, ORM1*S and ORM1*F2). One rare and one new allele were detected. The rare variant, tentatively assigned to the second structural locus ORM2, is observed in a cathodal position and named ORM2 B1. The new variant, tentatively assigned to the first structural locus ORM1, is observed in a region located between ORM1 S and ORM1 F2, and named ORM1 F3. Moreover, the pI values of the ORM variants have been measured accurately with Immobiline Dry Plates (LKB): they were found to be within the pH range 4.93-5.14.

Insertion of a malE B-Galactosidase fusion protein into the envelope of Escherichia coli disrupts biogenesis of outer membrane proteins and processing of inner membrane proteins

The synthesis of a membrane-bound MalE ,B-galactosidase hybrid protein, when induced by growth of Escherichia coli on maltose, leads to inhibition of cell division and eventually a reduced rate of mass increase. In addition, the relative rate of synthesis of outer membrane proteins, but not that of inner membrane proteins, was reduced by about 50%o. Kinetic experiments demonstrated that this reduction coincided with the period of maximum synthesis of the hybrid protein (and another maltose-inducible protein, LamB). The accumulation of this abnormal protein in the envelope therefore appeared specifically to inhibit the synthesis, the assembly of outer membrane proteins, or both, indicating that the hybrid protein blocks some export site or causes the sequestration of some limiting factor(s) involved in the export process. Since the MalE protein is normally located in the periplasm, the results also suggest that the synthesis of periplasmic and outer membrane proteins may involve some steps in common. The reduced rate of synthesis of outer membrane proteins was also accompanied by the accumulation in the envelope of at least one outer membrane protein and at least two inner membrane proteins as higher-molecular-weight forms, indicating that processing (removal of the N-terminal signal sequence) was also disrupted by the presence of the hybrid protein. These results may indicate that the assembly of these membrane proteins is blocked at a relatively late step rather than at the level of primary recognition of some site by the signal sequence. In addition, the results suggest that some step common to the biogenesis of quite different kinds of envelope protein is blocked by the presence of the hybrid protein.

Atividade da celulase e β-galactosidase no estudo da firmeza da polpa de mamões 'golden' e 'gran golden'

O objetivo desse trabalho foi avaliar a ação das enzimas celulase e β-galactosidase em relação à perda de firmeza dessas cultivares de mamões 'Gran Golden' e 'Golden' devido a relatos de uma perda de firmeza diferenciada entre as cvs. Os frutos foram armazenados a 25ºC e analisados diariamente quanto à firmeza da polpa e à atividade enzimática da celulase e β-galactosidase durante 8 dias. Os resultados de firmeza da polpa e atividade enzimática foram submetidos às análises de correlação e regressão. No 4º dia pós-colheita os mamões 'Golden' apresentaram firmeza média de 60,6 N e os 'Gran Golden' 31,1 N e a um aumento da atividade da celulase e da β-galactosidase. Os dados gerados neste trabalho sugerem que as enzimas celulase e β-galactosidase atuam diferentemente no processo de perda de firmeza dos frutos das cultivares Goldene Gran Golden. Aantecipaçãonaperdade firmezade 'Gran Golden' pode estar relacionada com a maior atividade dessas enzimas.

Preparation of alpha-labeled aldehydes by base-catalyzed exchange reactions

A simple, efficient protocol for the preparation of α-labeled aldehydes based on H/D exchange catalyzed by 4-(N,N-dimethylamino)pyridine or Et3N is described. High chemical yields and ratios of isotope incorporation were obtained even when small amounts (1 mmol) of aldehyde were used.

Preparation of alpha-labeled aldehydes by base-catalyzed exchange reactions

A simple, efficient protocol for the preparation of α-labeled aldehydes based on H/D exchange catalyzed by 4-(N,N-dimethylamino)pyridine or Et3N is described. High chemical yields and ratios of isotope incorporation were obtained even when small amounts (1 mmol) of aldehyde were used.

Atividade da pectina metilesterase e da β-galactosidase durante o amadurecimento do mamão cv. golden

Este trabalho foi realizado com o objetivo de investigar o comportamento das enzimas pectina metilesterase (PME) e β-galactosidase (β-Gal) durante o amadurecimento do mamão cv.'Golden'. Mamões com 15% de cor amarela foram estocados em câmara de refrigeração a 20ºC e 85-95 % UR, por um período de 11 dias. Durante este período de estocagem, foi determinada a cor da casca, firmeza e pH da polpa dos frutos, bem como a atividade das enzimas PME e β-Gal. A atividade da PME, observada ao longo do amadurecimento do mamão, indicou que esta enzima participou do processo de hidrólise da parede celular, principalmente no início do amadurecimento dos frutos. Já a atividade da β-Gal aumentou, de maneira não gradual, do primeiro dia até o final do armazenamento, quando alcançou seu valor máximo. Uma redução drástica na firmeza e no pH da polpa ocorreu nos três primeiros dias de armazenamento. Neste período, os frutos apresentaram coloração com valores do ângulo hue próximos de 80, a qual corresponde a uma coloração amarela. Verificou-se que a PME atua de modo efetivo no amadurecimento do mamão, mas a redução precoce em sua atividade indica que a β-Gal tem também um papel fundamental na rápida perda de firmeza pelo mamão cv. Golden.

Hepatocyte nuclear factor-4 alpha regulates the human apolipoprotein AV gene: Identification of a novel response element and involvement in the control by peroxisome proliferator-activated receptor-gamma coactivator-1 alpha, AMP-activated protein kinase, and mitogen-activated protein kinase pathway

The recently discovered apolipoprotein AV (apoAV) gene has been reported to be a key player in modulating plasma triglyceride levels. Here we identify the hepatocyte nuclear factor-4 (HNF-4 ) as a novel regulator of human apoAV gene. Inhibition of HNF-4 expression by small interfering RNA resulted in down-regulation of apoAV. Deletion, mutagenesis, and binding assays revealed that HNF-4 directly regulates human apoAV promoter through DR1 [a direct repeat separated by one nucleotide (nt)], and via a novel element for HNF-4 consisting of an inverted repeat separated by 8 nt (IR8). In addition, we show that the coactivator peroxisome proliferator-activated receptor- coactivator-1 was capable of stimulating the HNF-4 -dependent transactivation of apoAV promoter. Furthermore, analyses in human hepatic cells demonstrated that AMP-activated protein kinase (AMPK) and the MAPK signaling pathway regulate human apoAV expression and suggested that this regulation may be mediated, at least in part, by changes in HNF-4 . Intriguingly, EMSAs and mice with a liver-specific disruption of the HNF-4 gene revealed a species-distinct regulation of apoAV by HNF-4 , which resembles that of a subset of HNF-4 target genes. Taken together, our data provide new insights into the binding properties and the modulation of HNF-4 and underscore the role of HNF-4 in regulating triglyceride metabolism.

Avaliação da atividade das enzimas pectina metilesterase e β-Galactosidase em mamões cv. Golden armazenados sob diferentes concentrações de oxigênio

Este trabalho foi realizado com o objetivo de avaliar o efeito de atmosferas controladas contendo diferentes concentrações de oxigênio sobre a atividade das enzimas β-galactosidase e pectina metilesterase, e sobre a cor da casca e a firmeza da polpa de mamões 'Golden'. Os frutos foram mantidos por 36 dias, nas seguintes atmosferas controladas: 1% de O2 e 0,03% CO2 com adsorvedor de etileno, 3% de O2 e 0,03% de CO2 com adsorvedor de etileno, 5% O2 e 0,03% de CO2 com adsorvedor de etileno e atmosfera ambiente sem adsorvedor de etileno. A UR e a temperatura foram mantidas entre 85-95% e a 13º C, respectivamente. Os frutos estocados sob atmosfera de 1% de O2 e 0,03% CO2 apresentaram retardamento nas atividades das enzimas β-galactosidase e pectina metilesterase comparado com os frutos estocados nas outras atmosferas avaliadas. Os frutos armazenados sob atmosfera de 1% de O2 e 0,03% O2 também apresentaram atraso no desenvolvimento da cor da casca e amolecimento da polpa.