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1.
J Synchrotron Radiat ; 26(Pt 5): 1694-1704, 2019 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-31490161

RESUMEN

Corvus, a Python-based package designed for managing workflows of physical simulations that utilize multiple scientific software packages, is presented. Corvus can be run as an executable script with an input file and automatically generated or custom workflows, or interactively, in order to build custom workflows with a set of Corvus-specific tools. Several prototypical examples are presented that link density functional, vibrational and X-ray spectroscopy software packages and are of interest to the synchrotron community. These examples highlight the simplification of complex spectroscopy calculations that were previously limited to expert users, and demonstrate the flexibility of the Corvus infrastructure to tackle more general problems in other research areas.

2.
Appetite ; 121: 119-128, 2018 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-29104107

RESUMEN

This study investigated people's preferences for different water sources and factors that predict such preferences using a blind taste test. Water preferences of 143 participants for one name-brand bottled water, one groundwater-sourced tap water, and one indirect potable reuse (IDR) water were assessed. For predictors of water preference, we measured each participant's PTC taste sensitivity and assessed two personality traits (Neuroticism, Openness to Experience). We also explored participants' descriptions of each water source. Results indicate a preference for water treated with Reverse Osmosis (RO) (bottled and IDR water) over groundwater-sourced water, which had higher pH levels and lower concentrations of Ca and HCO3-. PTC taste sensitivity did not predict preferences, while Openness to Experience and Neuroticism predicted preference for IDR water. Positive relations between Openness to Experience and preferences for bottled and IDR water were moderated by gender and were stronger among females. Participants described water primarily by its taste and texture. Findings suggest that (1) tap water treated by RO is equally preferable to some bottled water, (2) personality traits may affect water preferences, and (3) prior findings of gender differences in preferences for bottled water may reflect personality characteristics. Efforts to increase acceptance for sustainable water alternatives, such as IDR, may be more successful by assuring consumers about taste and addressing personality traits that encourage or inhibit use.


Asunto(s)
Comportamiento del Consumidor , Agua Potable , Personalidad , Feniltiourea/análisis , Gusto , Adolescente , Adulto , Color , Femenino , Preferencias Alimentarias , Humanos , Masculino , Sensibilidad y Especificidad , Factores Sexuales , Olfato , Encuestas y Cuestionarios , Temperatura , Adulto Joven
3.
Environ Eng Sci ; 34(12): 853, 2017 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-29238157
4.
J Colloid Interface Sci ; 561: 849-853, 2020 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-31771871

RESUMEN

In this investigation, the utility of a static light scattering (SLS) technique to characterize aggregate morphology of two-dimensional engineered nanomaterials (2D ENMs) was systematically evaluated. The aggregation of graphene oxide (GO) and lithiated-molybdenum disulfide (Li-MoS2) were measured and compared to that of a spherical reference colloid, carboxylate-modified latex (CML) nanoparticles. The critical coagulation concentration (CCC) for all dispersions was determined via analysis of aggregation kinetics using time-resolved dynamic light scattering. This technique allowed for the elucidation of the transition from the reaction-limited aggregation (RLA) regime to diffusion-limited aggregation (DLA). The findings of this study support the aggregation trends predicted by Derjaguin-Landau-Verwey-Overbeek (DLVO) theory and recent computer simulations of aggregation kinetics. For all nanomaterials, as ionic strength increased towards the respective the CCC, fractal dimension decreased; any increase in ionic strength beyond the CCC did not yield significant change in fractal dimension. Across comparable primary particle sizes and using both carbonaceous (GO) and inorganic (Li-MoS2) 2D ENMs, this study further supports the use of SLS for the measurement of fractal dimension for 2D materials. To further support this claim, the aggregate morphology of GO in both RLA and DLA regimes was measured via cryogenic transmission electron microscopy.

5.
Environ Pollut ; 228: 35-42, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-28511037

RESUMEN

Increased use of commercial titanium dioxide nanoparticles (TiO2 NPs) in consumer products most likely leads to their additional environmental release. Aggregation and disaggregation processes are expected to play an important role in the fate and transport of TiO2 NPs in natural aquatic ecosystems. Therefore, in this work, we have studied the colloidal stability of TiO2 NPs in the presence of extracellular polymeric substances (EPS) from Bacillus subtilis and the adsorption behavior of EPS on TiO2 NPs in aqueous solutions at different pH values and ionic strengths (IS). The adsorption and aggregation processes were found to depend on the solution chemistry. The mass fraction of EPS on TiO2 NPs decreased with increased pH and NaCl concentrations, which was verified by Fourier transform infrared spectroscopy. The presence of EPS can substantially influence the colloidal stability of TiO2 NPs. In deionized water, the aggregation of NPs was induced by the addition of EPS only when the pH was below the TiO2 NP point of zero charge (≈6). When the pH was equal to pHPZC, TiO2, the TiO2 NPs would rapidly form large aggregates, but the adsorption of EPS leads to partial fragmentation via electrostatic repulsion and steric hindrance. When the pH was greater than pHPZC, TiO2, the aggregation rate was minimally affected by the increased EPS concentration. In NaCl solution, the aggregation rate of TiO2 NPs obviously increased with increased NaCl concentration. The critical coagulation concentration (CCC) of TiO2 NPs is 13.9 mM in the absence of EPS and increases to 155.6, 213.7 and 316.4 mM in the presence of 1, 5 and 10 mg/L EPS in NaCl solution, respectively, which indicates that the steric hindrance occurs after the addition of EPS. This study suggests that environmental conditions and EPS concentration greatly modify the colloidal stability of TiO2 nanoparticles.


Asunto(s)
Nanopartículas/química , Titanio/química , Contaminantes Químicos del Agua/química , Adsorción , Bacillus subtilis , Concentración de Iones de Hidrógeno , Concentración Osmolar , Polímeros , Electricidad Estática , Agua/química
6.
Water Res ; 104: 381-388, 2016 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-27576157

RESUMEN

The early stage of aggregation of titanium oxide (TiO2) nanoparticles was investigated in the presence of extracellular polymeric substance (EPS) constituents and common monovalent and divalent electrolytes through time-resolved dynamic light scattering (DLS). The hydrodynamic diameter was measured and the subsequent aggregation kinetics and attachment efficiencies were calculated across a range of 1-500 mM NaCl and 0.05-40 mM CaCl2 solutions. TiO2 particles were significantly aggregated in the tested range of monovalent and divalent electrolyte concentrations. The aggregation behavior of TiO2 particles in electrolyte solutions was in excellent agreement with the predictions based on Derjaguin-Landau-Verwey-Overbeek (DLVO) theory. Divalent electrolytes were more efficient in destabilizing TiO2 particles, as indicated by the considerably lower critical coagulation concentrations (CCC) (1.3 mM CaCl2 vs 11 mM NaCl). The addition of EPS to the NaCl and low concentration CaCl2 (0.05-10 mM) solutions resulted in a dramatic decrease in the aggregation rate and an increase in the CCC values. For solutions of 11 mM NaCl (the CCC values of TiO2 in the absence of EPS) and above, the resulting attachment efficiency was less than one, suggesting that the adsorbed EPS on the TiO2 nanoparticles led to steric repulsion, which effectively stabilized the nanoparticle suspension. At high CaCl2 concentrations (10-40 mM), however, the presence of EPS increased the aggregation rate. This is attributed to the aggregation of the dissolved extracellular polymeric macromolecules via intermolecular bridging, which in turn linked the TiO2 nanoparticles and aggregates together, resulting in enhanced aggregate growth. These results have important implications for assessing the fate and transport of TiO2 nanomaterials released in aquatic environments.


Asunto(s)
Electrólitos , Nanopartículas , Adsorción , Cinética , Polímeros
7.
Gene ; 260(1-2): 155-69, 2000 Dec 30.
Artículo en Inglés | MEDLINE | ID: mdl-11137301

RESUMEN

Sphingomonas paucimobilis var. EPA505 utilizes fluoranthene (FLA), naphthalene (NAP), and phenanthrene (PHE) as sole carbon sources for energy and growth. A genetic library of EPA505 was constructed using mini-Tn5 promoter reporter genes encoding for tetracycline resistance (tc(p-)) or luminescence (luxAB(p-)). Out of 2250 Tn5 mutants, ten were deficient in utilization of FLA, NAP, and/or PHE as sole carbon sources. Three classes of Tn5 mutants were defined: classI (nap(-)phe(-)fla(-)), classII (nap(-)phe(-)), and classIII (fla(-)). Four of five mutants in classI did not express dioxygenase function, whereas one classI mutant and all classII and classIII mutants retained dioxygenase activity. In Tn5 tc(p-) classI mutants 200 and 394 (dioxygenase negative) and classII mutant 132 (dioxygenase positive), promoter reporter was expressed when induced with FLA, NAP, PHE, other polycyclic aromatic hydrocarbons (PAHs), and several proposed PAH-derived catabolites. The Tn5 tc(p-) derived classIII mutant 104 was induced only with PAHs and not with PAH-derived catabolites. DNA sequence analysis of cloned regions of classI mutant 200 revealed that Tn5 inserted into a gene that shared (96%) DNA sequence homology with 2,3-dihydroxybiphenyl 1,2-dioxygenase that is designated pbhA. Nucleotide sequences downstream of pbhA shared (84%) homology to a Rieske-type ferredoxin subunit gene of a multicomponent dioxygenase designated pbhB. The Tn5 tc(p-) in classII mutant 132 occurred within sequences that shared (74%) homology with a trans-o-hydroxybenzylidene-pyruvate hydratase-aldolase gene (pbhC). Sequence analysis of the region proximal to this gene revealed a putative promoter that contained a binding site for a LysR transcriptional activator. In classIII mutant 104, the Tn5 tc(p-) resided within a region that shared 94% nucleotide homology to that of a pyruvate phosphate dikinase gene known to be involved in cellular uptake of glucose. The FLA-specific catabolic gene disrupted in mutant 104 was designated phbD. Functional and sequence analyses of promoter probe mutants allowed identification of four genes necessary for the utilization of PAHs that are controlled by at least two promoters that are affected by a wide range of aromatic compounds.


Asunto(s)
Dioxigenasas , Genes Bacterianos/genética , Hidrocarburos Policíclicos Aromáticos/metabolismo , Regiones Promotoras Genéticas/genética , Sphingomonas/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Secuencia de Bases , División Celular/efectos de los fármacos , División Celular/genética , Medios de Cultivo/farmacología , Elementos Transponibles de ADN/genética , ADN Bacteriano/química , ADN Bacteriano/genética , Fluorenos/metabolismo , Fluorenos/farmacología , Hidrocarburos Aromáticos/farmacología , Datos de Secuencia Molecular , Mutagénesis Insercional , Mutación , Naftalenos/metabolismo , Naftalenos/farmacología , Oxigenasas/metabolismo , Fenantrenos/metabolismo , Fenantrenos/farmacología , Hidrocarburos Policíclicos Aromáticos/farmacología , Piruvato Quinasa/metabolismo , Piruvato Ortofosfato Diquinasa/metabolismo , Análisis de Secuencia de ADN , Sphingomonas/efectos de los fármacos , Sphingomonas/metabolismo
8.
J Med Chem ; 35(23): 4330-3, 1992 Nov 13.
Artículo en Inglés | MEDLINE | ID: mdl-1360025

RESUMEN

A series of dynorphin A-(1-13) amide (Dyn A-(1-13)NH2) analogues containing lysine or N epsilon-acetyllysine (Lys(Ac)) was prepared by solid-phase peptide synthesis and evaluated for opioid receptor affinity in radioligand binding assays and for opioid activity in the guinea pig ileum (GPI). Substitutions were made at positions 6, 7, 9, 11, and 13, the basic amino acids in the C-terminus of the peptide, in order to assess the individual contributions of these residues to the kappa opioid receptor affinity and selectivity of Dyn A-(1-13)NH2. While substitutions of Lys(Ac) for Arg in position 6 did not affect kappa receptor affinity, it enhanced affinity for mu and delta receptors and therefore caused a loss of kappa receptor selectivity. When Lys(Ac) was substituted for Arg9, kappa opioid receptor affinity was enhanced and kappa receptor selectivity was retained. Replacement for Arg7, Lys11, or Lys13 by Lys(Ac) resulted in both decreased affinity and selectivity for kappa receptors. These results demonstrate the importance of Arg6 to the receptor selectivity profile of Dyn A-(1-13)NH2 and indicate that, of the five basic residues in the C-terminus, only Arg9 can be replaced by a nonbasic residue without substantial loss of kappa opioid receptor selectivity.


Asunto(s)
Dinorfinas/análogos & derivados , Fragmentos de Péptidos/síntesis química , Secuencia de Aminoácidos , Aminoácidos/análisis , Animales , Cromatografía Líquida de Alta Presión , Cobayas , Datos de Secuencia Molecular , Músculo Liso/efectos de los fármacos , Fragmentos de Péptidos/metabolismo , Fragmentos de Péptidos/farmacología , Receptores Opioides/efectos de los fármacos , Receptores Opioides/metabolismo , Relación Estructura-Actividad
9.
J Appl Physiol (1985) ; 87(3): 1154-62, 1999 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-10484590

RESUMEN

We examined gender differences in growth hormone (GH) secretion during rest and exercise. Eighteen subjects (9 women and 9 men) were tested on two occasions each [resting condition (R) and exercise condition (Ex)]. Blood was sampled at 10-min intervals from 0600 to 1200 and was assayed for GH by chemiluminescence. At R, women had a 3.69-fold greater mean calculated mass of GH secreted per burst compared with men (5.4 +/- 1.0 vs. 1.7 +/- 0.4 microg/l, respectively) and higher basal (interpulse) GH secretion rates, which resulted in greater GH production rates and serum GH area under the curve (AUC; 1,107 +/- 194 vs. 595 +/- 146 microg x l(-1) x min, women vs. men; P = 0.04). Compared with R, Ex resulted in greater mean mass of GH secreted per burst, greater mean GH secretory burst amplitude, and greater GH AUC (1,196 +/- 211 vs. 506 +/- 90 microg x l(-1) x min, Ex vs. R, respectively; P < 0.001). During Ex, women attained maximal serum GH concentrations significantly earlier than men (24 vs. 32 min after initiation of Ex, respectively; P = 0.004). Despite this temporal disparity, both genders had similar maximal serum GH concentrations. The change in AUC (adjusted for unequal baselines) was similar for men and women (593 +/- 201 vs. 811 +/- 268 microg x l(-1) x min), but there were significant gender-by-condition interactive effects on GH secretory burst mass, pulsatile GH production rate, and maximal serum GH concentration. We conclude that, although women exhibit greater absolute GH secretion rates than men both at rest and during exercise, exercise evokes a similar incremental GH response in men and women. Thus the magnitude of the incremental secretory GH response is not gender dependent.


Asunto(s)
Ejercicio Físico/fisiología , Hormona de Crecimiento Humana/metabolismo , Adulto , Umbral Anaerobio/fisiología , Área Bajo la Curva , Composición Corporal/fisiología , Femenino , Hormonas Esteroides Gonadales/sangre , Humanos , Factor I del Crecimiento Similar a la Insulina/metabolismo , Ácido Láctico/sangre , Masculino , Consumo de Oxígeno/fisiología , Caracteres Sexuales
10.
Int J Pept Protein Res ; 43(3): 292-6, 1994 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-8005752

RESUMEN

The coupling reagent 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) was used in an attempt to prepare a highly strained 10-membered lactam ring on a solid support via side-chain to side-chain cyclization of the adjacent alpha,gamma-diaminobutyric (Dab) and D-glutamic acid residues in [Dab2,D-Glu3,Leu5]enkephalinamide. This attempted cyclization failed, however, and yielded linear products instead. Characterization by mass spectrometry, amino acid analysis, peptide sequencing and NMR indicated that the major products were the tetramethylguanidinium (Tmg) derivatives [Dab(Tmg)2,D-Glu3,Leu5]-enkephalinamide and the corresponding dimeric linear Tmg-containing peptide which resulted from the transfer of the tetramethyluronium moiety from HBTU to the amino side chain of Dab. The formation of these tetramethylguanidinium side products during cyclization reactions limits HBTU's usefulness for the formation of lactams.


Asunto(s)
Péptidos Cíclicos/síntesis química , Triazoles , Urea/análogos & derivados , Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Encefalina Leucina/análogos & derivados , Encefalina Leucina/síntesis química , Datos de Secuencia Molecular , Conformación Proteica
11.
Int J Pept Protein Res ; 39(1): 87-92, 1992 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-1353068

RESUMEN

Different resins were examined for their potential use in the solid phase synthesis of protected peptide amides using the 9-fluorenylmethoxycarbonyl (Fmoc) chemical protocol. The model protected peptide amide BocTyr-Gly-Gly-Phe-Leu-Arg(Pmc)NH2 (1) was synthesized on both the acid-labile 4-(2',4'-dimethoxyphenyl-Fmoc-aminomethyl)phenoxy resin (Rink amide resin) (2) and on resins containing the base-labile linker 4-hydroxymethylbenzoic acid. Of the resins examined only the methylbenzhydrylamine resin containing the 4-hydroxymethylbenzoic acid linkage, which was cleaved by ammonolysis in isopropanol, gave the model peptide 1 in good overall yield (53% including functionalization). Thus the synthesis of protected peptide amides by solid phase synthesis using Fmoc-protected amino acids with t-butyl-type side chain protecting groups is feasible. The choice of peptide-resin linkage and its cleavage conditions, however, are critical to the success of such syntheses. The potential application of this synthetic strategy to the preparation of novel peptide amides is discussed.


Asunto(s)
Compuestos de Bencidrilo , Fluorenos , Oligopéptidos/síntesis química , Resinas Sintéticas , Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Dinorfinas/química , Datos de Secuencia Molecular , Oligopéptidos/química , Espectrometría de Masa Bombardeada por Átomos Veloces
12.
J Bacteriol ; 183(14): 4259-68, 2001 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-11418567

RESUMEN

Aminoacylase was identified in cell extracts of the hyperthermophilic archaeon Pyrococcus furiosus by its ability to hydrolyze N-acetyl-L-methionine and was purified by multistep chromatography. The enzyme is a homotetramer (42.06 kDa per subunit) and, as purified, contains 1.0 +/- 0.48 g-atoms of zinc per subunit. Treatment of the purified enzyme with EDTA resulted in complete loss of activity. This was restored to 86% of the original value (200 U/mg) by treatment with ZnCl(2) (and to 74% by the addition of CoCl(2)). After reconstitution with ZnCl(2), the enzyme contained 2.85 +/- 0.48 g-atoms of zinc per subunit. Aminoacylase showed broad substrate specificity and hydrolyzed nonpolar N-acylated L amino acids (Met, Ala, Val, and Leu), as well as N-formyl-L-methionine. The high K(m) values for these compounds indicate that the enzyme plays a role in the metabolism of protein growth substrates rather than in the degradation of cellular proteins. Maximal aminoacylase activity with N-acetyl-L-methionine as the substrate occurred at pH 6.5 and a temperature of 100 degrees C. The N-terminal amino acid sequence of the purified aminoacylase was used to identify, in the P. furiosus genome database, a gene that encodes 383 amino acids. The gene was cloned and expressed in Escherichia coli by using two approaches. One involved the T7 lac promoter system, in which the recombinant protein was expressed as inclusion bodies. The second approach used the Trx fusion system, and this produced soluble but inactive recombinant protein. Renaturation and reconstitution experiments with Zn(2+) ions failed to produce catalytically active protein. A survey of databases showed that, in general, organisms that contain a homolog of the P. furiosus aminoacylase (> or = 50% sequence identity) utilize peptide growth substrates, whereas those that do not contain the enzyme are not known to be proteolytic, suggesting a role for the enzyme in primary catabolism.


Asunto(s)
Amidohidrolasas/metabolismo , Pyrococcus furiosus/enzimología , Amidohidrolasas/genética , Amidohidrolasas/aislamiento & purificación , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Catálisis , ADN de Archaea , Datos de Secuencia Molecular , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/aislamiento & purificación , Proteínas Recombinantes de Fusión/metabolismo , Homología de Secuencia de Aminoácido , Porcinos
13.
Int J Pept Protein Res ; 40(2): 89-96, 1992 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-1359999

RESUMEN

A series of 2-substituted dynorphin A-(1-13) amide (Dyn A-(1-13)NH2) analogues was prepared by solid phase peptide synthesis and evaluated for opioid receptor affinities in radioligand binding assays and for opioid activity in the guinea pig ileum (GPI) assay. Amino acid substitution at the 2 position produced marked differences in both opioid receptor affinities and potency in the GPI assay; Ki values for the analogues in the radioligand binding assays and IC50 values in the GPI assay varied over three to four orders of magnitude. The parent peptide, Dyn A-(1-13)NH2, exhibited the greatest affinity and selectivity for kappa receptors and was the most potent peptide examined in the GPI assay. The most important determinant of opioid receptor selectivity and opioid potency for the synthetic analogues was the stereochemistry of the amino acid at the 2 position. Except for [D-Lys2]Dyn A-(1-13)NH2 in the kappa receptor binding assay, the analogues containing a D-amino acid at position 2 were much more potent in all of the assays than their corresponding isomers containing an L-amino acid at this position. The L-amino acid-substituted analogues generally retained some selectivity for kappa opioid receptors. The more potent derivatives with a D-amino acid in position 2, however, preferentially interacted with mu opioid receptors. Introduction of a positively charged amino acid into the 2 position generally decreased opioid receptor affinities and potency in the GPI assay.


Asunto(s)
Dinorfinas/farmacología , Endorfinas/farmacología , Narcóticos/farmacología , Fragmentos de Péptidos/farmacología , Amidas/síntesis química , Amidas/metabolismo , Amidas/farmacología , Secuencia de Aminoácidos , Animales , Dinorfinas/síntesis química , Dinorfinas/metabolismo , Endorfinas/síntesis química , Endorfinas/metabolismo , Cobayas , Técnicas In Vitro , Cinética , Datos de Secuencia Molecular , Narcóticos/síntesis química , Narcóticos/metabolismo , Fragmentos de Péptidos/síntesis química , Fragmentos de Péptidos/metabolismo , Ensayo de Unión Radioligante , Ratas , Receptores Opioides/metabolismo , Relación Estructura-Actividad
14.
Arch Environ Contam Toxicol ; 47(2): 168-76, 2004 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-15386141

RESUMEN

To determine the substrate range capability of Sphingomonas paucimobilis strain EPA505, a number of aromatic compounds were tested as potential growth substrates. Strain EPA505 grew on phenanthrene, naphthalene, fluoranthene, toluene, benzoic acid, 2,3- and 3,4-dihydroxybenzoic acids, 1-chloro-2,4-dinitrobenzene, anthracene, 2-hydroxy-3-naphthoic acid and 1-hydroxy- 2-naphthoic acid, salicylic acid, and catechol. Strain EPA505 was unable to grow on coumarine 3-carboxylic acid, naphthalene dicarboxylic acid, acenaphthene, chrysene, pyrene, benzo[b]fluoranthene, and fluorene. Catabolic products were not detected or identified when the bacterium was incubated with coumarine 3-carboxylic acid, naphthalene dicarboxylic acid, acenaphthene, chrysene, or benzo[b]fluoranthene. Dihydroxypyrene, the ortho ring fission product of pyrene, and 10-hydroxy-1- phenanthroic acid were detected when the bacterium was incubated with pyrene. The open rings of benzo[b]fluoranthene, hydroxyacephenanthroic acid, hydroxyacephenanthrene, and phenanthrene anhydride, catabolites of benzo[b]fluoranthene degradation, were detected with Tn5 mutants of EPA505. With strain EPA505, both 9-fluorenone and an open ring fission product accumulated during incubation with fluorene. Other catabolites beyond the open ring of fluorene were detected, specifically dihydroxyfluorene, hydroxy-9-fluorenone, dihydroxy-9-fluorenone, hydroxyindane, and a putative glutathione-conjugated benzylanhydride. Benzylanhydride appeared to be a final end product of fluorene degradation by strain EPA505.


Asunto(s)
Hidrocarburos Policíclicos Aromáticos/metabolismo , Sphingomonas/fisiología , Contaminantes Químicos del Agua/metabolismo , Peso Molecular , Hidrocarburos Policíclicos Aromáticos/toxicidad , Sphingomonas/crecimiento & desarrollo , Contaminantes Químicos del Agua/toxicidad
15.
J Ind Microbiol Biotechnol ; 26(6): 369-82, 2001 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-11571621

RESUMEN

Catabolic pathways for utilization of naphthalene (NAP), anthracene (ANT), phenanthrene (PHE), and fluoranthene (FLA) by Sphingomonas paucimobilis EPA505 were identified. Accumulation of catabolic intermediates was investigated with three classes of Tn5 mutants with the following polycyclic aromatic hydrocarbon (PAH)-negative phenotypes; (class I NAP(-) PHE(-) FLA(-), class II NAP(-) PHE(-), and class III FLA(-)). Class I mutant 200pbhA had a Tn5 insertion within a meta ring fission dioxygenase (pbhA), and a ferredoxin subunit gene (pbhB) resided directly downstream. Mutant 200pbhA and other class I mutants lost the ability to catalyze the initial dihydroxylation step and did not transform NAP, ANT, PHE, or FLA. Class I mutant 401 accumulated salicylic acid, 2-hydroxy-3-naphthoic acid, 1-hydroxy-2-naphthoic acid, and hydroxyacenaphthoic acid during incubation with NAP, ANT, PHE, or FLA, respectively. Class II mutant 132pbhC contained the Tn5 insertion in an aldolase hydratase (pbhC) and accumulated what appeared to be meta ring fission products: trans-o-hydroxybenzylidene pyruvate, trans-o-hydroxynaphylidene pyruvate, and trans-o-hydroxynaphthyl-oxobutenoic acid when incubated with NAP, ANT, and PHE, respectively. When mutant 132pbhC was incubated with 1-hydroxy-2-naphthoic acid, it accumulated trans-o-hydroxybenzylidene pyruvate. Class III mutant 104ppdk had a Tn5 insertion in a pyruvate phosphate dikinase gene that affected expression of a FLA-specific gene and accumulated a proposed meta ring fission product; trans-o-hydroxyacenaphyl-oxobutenoic acid during incubation with FLA. Trans-o-hydroxyacenaphyl-oxobutenoic acid was degraded to acenaphthenone that accumulated with class III mutant 611. Acenaphthenone was oxidized via incorporation of one molecule of dioxygen by another oxygenase. 2,3-Dihydroxybenzoic acid was the final FLA-derived catabolic intermediate detected. Analysis of PAH utilization mutants revealed that there are convergent and divergent points involved in NAP, ANT, PHE, and FLA utilization by S. paucimobilis EPA505.


Asunto(s)
Antracenos/metabolismo , Fluorenos/metabolismo , Naftalenos/metabolismo , Fenantrenos/metabolismo , Sphingomonas/genética , Sphingomonas/metabolismo , Cromatografía Líquida de Alta Presión , Oxigenasas de Función Mixta/genética , Oxigenasas de Función Mixta/metabolismo , Mutación/genética , Operón/genética , Oxigenasas/genética , Oxigenasas/metabolismo , Sphingomonas/clasificación , Sphingomonas/enzimología
16.
Am J Physiol Regul Integr Comp Physiol ; 279(4): R1455-66, 2000 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-11004016

RESUMEN

We test the hypotheses that 1) growth hormone (GH)-releasing peptide-2 (G) synergizes with L-arginine (A), a compound putatively achieving selective somatostatin withdrawal and 2) gender modulates this synergy on GH secretion. To these ends, 18 young healthy volunteers (9 men and 9 early follicular phase women) each received separate morning intravenous infusions of saline (S) or A (30 g over 30 min) or G (1 microg/kg) or both, in randomly assigned order. Blood was sampled at 10-min intervals for later chemiluminescence assay of serum GH concentrations. Analysis of covariance revealed that the preinjection (basal) serum GH concentrations significantly determined secretagogue responsiveness and that sex (P = 0.02) and stimulus type (P < 0.001) determined the slope of this relationship. Nested ANOVA applied to log-transformed measures of GH release showed that gender determines 1) basal rates of GH secretion, 2) the magnitude of the GH secretory response to A, 3) the rapidity of attaining the GH maximum, and 4) the magnitude or fold (but not absolute) elevation in GH secretion above preinjection basal, as driven by the combination of A and G. In contrast, the emergence of the G and A synergy is sex independent. We conclude that gender modulates key facets of basal and A/G-stimulated GH secretion in young adults.


Asunto(s)
Arginina/farmacología , Hormona de Crecimiento Humana/metabolismo , Oligopéptidos/farmacología , Caracteres Sexuales , Ciclos de Actividad/efectos de los fármacos , Adulto , Análisis de Varianza , Femenino , Semivida , Hormonas/farmacología , Hormona de Crecimiento Humana/sangre , Hormona de Crecimiento Humana/farmacología , Humanos , Mediciones Luminiscentes , Masculino , Modelos Biológicos , Modelos Estadísticos
17.
Am J Physiol Regul Integr Comp Physiol ; 279(4): R1467-77, 2000 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-11004017

RESUMEN

We investigated the ability of exercise, a multipathway, potent, physiological stimulus for GH release, to alter the synergistic interaction of L-arginine (A) and GH-related peptide (GHRP)-2 (G) observed at rest and the ability of gender to further modulate this putative interaction. Subjects (9 men and 9 early follicular phase women) completed 30 min of constant load aerobic exercise in combination with intravenous infusions of saline (S), A (30 g over 30 min), G (1 microg/kg bolus), or both (AG) in separate study sessions in randomly assigned order. Measures of GH release were logarithmically transformed for statistical analysis. Similar to rest, exercise maintained the rank order (AG > G > A > S) of effective stimulation of GH release for the key response measures in men or women, a gender disparity in the time to reach the maximal serum GH concentration, the calculated endogenous GH half-life, and the observed effect of preinfusion (basal) serum GH concentrations on determining secretagogue responsiveness. Exercise potentiated the individual stimulatory actions of A and G, while blunting the relative magnitude of the synergistic (supra-additive) interaction observed at rest. We infer from the present data that 1) exercise is likely to induce release of both GHRH and somatostatin, 2) L-arginine may facilitate the effect of exercise by limiting somatostatin release, 3) GHRP-2 could further enhance the stimulatory impact of exercise by opposing central actions of somatostatin and/or heightening endogenous GHRH release, and 4) gender strongly controls the relative but not absolute magnitude of A/G synergy both at rest and after exercise.


Asunto(s)
Arginina/farmacología , Ejercicio Físico/fisiología , Hormonas/farmacología , Hormona de Crecimiento Humana/metabolismo , Oligopéptidos/farmacología , Esfuerzo Físico/fisiología , Adulto , Análisis de Varianza , Arginina/administración & dosificación , Sinergismo Farmacológico , Metabolismo Energético/efectos de los fármacos , Femenino , Fase Folicular , Hormonas/administración & dosificación , Hormona de Crecimiento Humana/sangre , Humanos , Infusiones Intravenosas , Masculino , Oligopéptidos/administración & dosificación , Consumo de Oxígeno/efectos de los fármacos , Análisis de Regresión , Caracteres Sexuales
18.
J Bacteriol ; 183(2): 716-24, 2001 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-11133967

RESUMEN

The hyperthermophilic archaeon Pyrococcus furiosus grows optimally at 100 degrees C by the fermentation of peptides and carbohydrates. Growth of the organism was examined in media containing either maltose, peptides (hydrolyzed casein), or both as the carbon source(s), each with and without elemental sulfur (S(0)). Growth rates were highest on media containing peptides and S(0), with or without maltose. Growth did not occur on the peptide medium without S(0). S(0) had no effect on growth rates in the maltose medium in the absence of peptides. Phenylacetate production rates (from phenylalanine fermentation) from cells grown in the peptide medium containing S(0) with or without maltose were the same, suggesting that S(0) is required for peptide utilization. The activities of 14 of 21 enzymes involved in or related to the fermentation pathways of P. furiosus were shown to be regulated under the five different growth conditions studied. The presence of S(0) in the growth media resulted in decreases in specific activities of two cytoplasmic hydrogenases (I and II) and of a membrane-bound hydrogenase, each by an order of magnitude. The primary S(0)-reducing enzyme in this organism and the mechanism of the S(0) dependence of peptide metabolism are not known. This study provides the first evidence for a highly regulated fermentation-based metabolism in P. furiosus and a significant regulatory role for elemental sulfur or its metabolites.


Asunto(s)
Hidrogenasas/metabolismo , Péptidos/metabolismo , Pyrococcus furiosus/metabolismo , Azufre/metabolismo , Medios de Cultivo , Citoplasma/enzimología , Regulación de la Expresión Génica Arqueal , Regulación Enzimológica de la Expresión Génica , Glucólisis , Proteínas de la Membrana/metabolismo , Oxidación-Reducción
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