Pi*-pi* bonding interactions generated by halogen oxidation of zirconium(IV) redox-active ligand complexes.

The new complex, [Zr(pda)2]n (1, pda2- = N,N'-bis(neo-pentyl)-ortho-phenylenediamide, n = 1 or 2), prepared by the reaction of 2 equiv of pdaLi2 with ZrCl4, reacts rapidly with halogen oxidants to afford the new product ZrX2(disq)2 (3, X = Cl, Br, I; disq- = N,N'-bis(neo-pentyl)-ortho-diiminosemiquinonate) in which each redox-active ligand has been oxidized by one electron. The oxidation products 3a-c have been structurally characterized and display an unusual parallel stacked arrangement of the disq- ligands in the solid state, with a separation of approximately 3 A. Density functional calculations show a bonding-type interaction between the SOMOs of the disq- ligands to form a unique HOMO while the antibonding linear combination forms a unique LUMO. This orbital configuration leads to a closed-shell-singlet ground-state electron configuration (S = 0). Temperature-dependent magnetism measurements indicate a low-lying triplet excited state at approximately 750 cm-1. In solution, 3a-c show strong disq--based absorption bands that are invariant across the halide series. Taken together these spectroscopic measurements provide experimental values for the one- and two-electron energies that characterize the pi-stacked bonding interaction between the two disq- ligands.

Pruebas preliminares sobre toxicidad aguda del cobre en la Concha de Abanico (Argopecten purpuratus)

Se realizaron siete bioensayos estáticos con la Concha de Abanico (Argopecten purpuratus) con concentraciones de cobre que variaron de 0.007 a 0.74 ppm. El agua de mar de donde provinieron los individuos usados en el experimento tenía concentraciones de cobre entre 0.005-0.007 ppm. La bioacumulación inicial en los animales varó de 1.69 a 6.50 ppm. Como resultado preliminar se determinó que 0.13 ppm es la concentración letal media (LC 50) en 96 horas.

In vivo characterization of sunitinib eluting beads

Purpose: To study the pharmacokinetics and potential toxicity of sunitinib eluting beads in an animal modelMaterials: Healthy New-Zealand white rabbits were used. 8 animals received 0.2ml of DC Beads loaded with 6mg of sunitinb intra arterially in the hepatic artery (group 1) and 4 animals received 6mg of sunitinib administered orally (group 2). In group 1, animals were sacrificed 6 hours (n=4) and 24 hours (n=4) after embolization. In group 2, animals were sacrificed 6 hours (n=2) and 24 hours (n=2) after oral administration of sunitinib. Liver enzymes were measured at 0, 6 and 24 hours in both groups. Plasmatic sunitinib concentration was measured by LC MS/MS tandem mass spectroscopy at 0, 1, 2, 3, 4, 5, 6 and 24 hours. At sacrifice, the livers were harvested and sunitinib concentration in liver tissue was assessed by LC MS/MS tandem mass spectroscopy.Results: After embolization we observed an expected elevation of AST and ALT. Serial plasmatic measurments after embolization showed a very low sunitinib concentration (<50ng/ml). Measurment of sunitinib in the embolized liver tissue showed a very high concentration at 6 hours (3870ng/ml) and 24 hours (4741.7ng/ml).Conclusions: Sunitinib eluting beads are well tolerated by rabbits when administered intra-arterially in the hepatic artery. No unexpected toxicity was observed. Very high drug concentration canbe obtained at the site of embolization with minimal systemic passage.

Food's visually perceived fat content affects discrimination speed in an orthogonal spatial task.

Choosing what to eat is a complex activity for humans. Determining a food's pleasantness requires us to combine information about what is available at a given time with knowledge of the food's palatability, texture, fat content, and other nutritional information. It has been suggested that humans may have an implicit knowledge of a food's fat content based on its appearance; Toepel et al. (Neuroimage 44:967-974, 2009) reported visual-evoked potential modulations after participants viewed images of high-energy, high-fat food (HF), as compared to viewing low-fat food (LF). In the present study, we investigated whether there are any immediate behavioural consequences of these modulations for human performance. HF, LF, or non-food (NF) images were used to exogenously direct participants' attention to either the left or the right. Next, participants made speeded elevation discrimination responses (up vs. down) to visual targets presented either above or below the midline (and at one of three stimulus onset asynchronies: 150, 300, or 450 ms). Participants responded significantly more rapidly following the presentation of a HF image than following the presentation of either LF or NF images, despite the fact that the identity of the images was entirely task-irrelevant. Similar results were found when comparing response speeds following images of high-carbohydrate (HC) food items to low-carbohydrate (LC) food items. These results support the view that people rapidly process (i.e. within a few hundred milliseconds) the fat/carbohydrate/energy value or, perhaps more generally, the pleasantness of food. Potentially as a result of HF/HC food items being more pleasant and thus having a higher incentive value, it seems as though seeing these foods results in a response readiness, or an overall alerting effect, in the human brain.

Quantitative monitoring of tamoxifen in human plasma extended to 40 metabolites using liquid-chromatography high-resolution mass spectrometry: new investigation capabilities for clinical pharmacology.

Liquid-chromatography (LC) high-resolution (HR) mass spectrometry (MS) analysis can record HR full scans, a technique of detection that shows comparable selectivity and sensitivity to ion transitions (SRM) performed with triple-quadrupole (TQ)-MS but that allows de facto determination of "all" ions including drug metabolites. This could be of potential utility in in vivo drug metabolism and pharmacovigilance studies in order to have a more comprehensive insight in drug biotransformation profile differences in patients. This simultaneous quantitative and qualitative (Quan/Qual) approach has been tested with 20 patients chronically treated with tamoxifen (TAM). The absolute quantification of TAM and three metabolites in plasma was realized using HR- and TQ-MS and compared. The same LC-HR-MS analysis allowed the identification and relative quantification of 37 additional TAM metabolites. A number of new metabolites were detected in patients' plasma including metabolites identified as didemethyl-trihydroxy-TAM-glucoside and didemethyl-tetrahydroxy-TAM-glucoside conjugates corresponding to TAM with six and seven biotransformation steps, respectively. Multivariate analysis allowed relevant patterns of metabolites and ratios to be associated with TAM administration and CYP2D6 genotype. Two hydroxylated metabolites, α-OH-TAM and 4'-OH-TAM, were newly identified as putative CYP2D6 substrates. The relative quantification was precise (<20 %), and the semiquantitative estimation suggests that metabolite levels are non-negligible. Metabolites could play an important role in drug toxicity, but their impact on drug-related side effects has been partially neglected due to the tremendous effort needed with previous MS technologies. Using present HR-MS, this situation should evolve with the straightforward determination of drug metabolites, enlarging the possibilities in studying inter- and intra-patients drug metabolism variability and related effects.

Compressibilidade de três Latossolos em função da umidade e uso

Por meio do ensaio de compressibilidade, foram estudados os efeitos do manejo e da umidade na pressão de preconsolidação (σp) de três solos: Latossolo Vermelho-Amarelo, Latossolo Roxo e Latossolo Vermelho-Escuro sob cultura anual, mata natural e pastagem, na região de Lavras (MG), em duas profundidades (0-0,03 e 0,27-0,30 m). Para cada condição, foram coletadas cinco amostras indeformadas e uma deformada, com três repetições. As amostras, indeformadas e com diferentes umidades, foram utilizadas no ensaio de compressão uniaxial, obtendo-se as curvas de compressão, das quais foram extraídas as respectivas pressões de preconsolidação. Com as amostras deformadas, determinaram-se os limites de plasticidade e de contração, textura e matéria orgânica. Os modelos de compactação testados foram baseados na pressão de preconsolidação e na umidade do solo. Para uma mesma condição e profundidade, houve diferença significativa entre os valores dos teores de argila e areia nos três solos. Os valores da densidade do solo inicial (Ds i) foram estatisticamente diferentes para todas as condições na mesma profundidade, exceto na camada de 0-0,03 m para a cultura anual. À medida que a umidade do solo aumentou, a pressão de preconsolidação decresceu exponencialmente, indicando uma redução na capacidade de suporte de carga do solo. O LR apresentou, em geral, maior capacidade de suporte de carga do que o LE e LV. Essa maior capacidade de suporte de carga pode estar associada com o seu maior teor de argila e menor teor de areia. A capacidade de suporte de carga na zona de friabilidade variou de 154 a 167 kPa, para o LV; de 77 a 183 kPa, para o LR, e de 77 a 132 kPa, para o LE. As umidades 0,33 a 0,30 kg kg-1, 0,42 a 0,27 kg kg-1, e 0,35 a 0,33 kg kg-1 correspondem à faixa de friabilidade (LP - LC) do LV, LR e LE, respectivamente. O modelo baseado na história de tensão do solo evidenciou o efeito da compactação causada pelas máquinas de preparo do solo na camada de 0,27-0,30 m, para a cultura anual, enquanto, para a pastagem, ocorreu o efeito do pisoteio do gado na camada de 0-0,03 m.

Aspectos biológico - pesqueros del Camaroncito Rojo Pleuroncodes monodon (M Edwards, 1837) en el litoral peruano

Se analizaron algunos parámetros poblacionales de 11 583 ejemplares del Pleuroncodes monodon (CRUSTACEA: ANOMURA: GALAYHEIDAE) distribuidos en 5 860 (52%) ejemplares machos y 5723 (48%) ejemplares hembras, las muestras se obtuvieron de los lances de comprobación de los cruceros de evaluación tanto para recursos pelágicos y demersales ejecutados por el Instituto del Mar del Perú (IMARPE) entre febrero y noviembre del 2001, la zona de estudio estuvo comprendida entre Puerto Pizarro (03°29,1’ S; 80°23,0’ W) hasta Los Palos (18°20,18’ S; 70°22,5’ W). La presencia del Pleuroncodes monodon respecto a su distribución batimétrica abarcó entre 1,5 y 300 m. De profundidad. La distribución total de tallas fluctuó entre 6,0 y 42,4 mm de longitud cefalotoráxica (Lc) y entre 6,0 y 41,1 mm de Lc para machos y hembras respectivamente. Por estaciones la estructura por tallas fue variable denotándose dos marcados grupos de talla, predominando un primer grupo los denominados ”juveniles” de (de 6 hasta 20 mm de Lc), sin embargo aquellos ejemplares con longitud cefalotoráxica menor a los 13 mm se registraron mayormente en las estaciones de verano y primavera, mientras un segundo grupo perteneciente a los de talla intermedia denominados “adultos jóvenes” registrados en todas las estaciones. Su distribución horizontal fue mayormente costera; aunque su presencia fue hasta más de las 50 millas náuticas (mn) de la costa; sin embargo los mayores registros de este recurso fue dentro de las 25 mn de la costa, en cuanto a los ejemplares “juveniles” su presencia fue marcada dentro de las 10 mn de la costa. La composición sexual de la población muestreada varió durante el periodo de estudio obteniéndose una razón promedio de 1,02 (machos/hembras). Además se encontraron hembras con huevos en todo el periodo de estudio, sin embargo los altos porcentajes de hembras ovígeras se registraron entre las estaciones de invierno (70%) y primavera (68%) del total. La talla de madurez sexual en las hembras fue de 18,8 mm Lc, mientras que el primer desove en 17,4 mm de Lc. Al registrarse hembras ovígeras en todo el año, existe un periodo anual de postura (liberación de los huevos eclosionados), detectándose éste en la estacón de primavera. La relación longitud cefalotoráxica vs Fecundidad, está determinada por la ecuación: NHT 0,1899 Lc2,9883 Además se analizaron los siguientes parámetros: largo cefalotoráxico, peso total, peso del abdomen, características de los huevos. Complementándose tales evaluaciones con determinaciones de áreas de distribución y concentración, así como la relación de los factores: abióticos (temperaturas, salinidad y oxígeno) y bióticos en relación a predadores y competidores dentro de la cadena trófica.

Therapeutic Drug Monitoring (TDM) of Imatinib: Effectiveness of Bayesian dose adjustment for CML patients enrolled in the Imatinib Concentration Monitoring (I-COME) study

Introduction: As imatinib pharmacokinetics are highly variable, plasma levels differ largely between patients under the same dosage. Retrospective studies in chronic myeloid leukemia (CML) patients showed significant correlations between low levels and suboptimal response, as well as between high levels and poor tolerability. Monitoring of trough plasma levels, targeting 1000 μg/L and above, is thus increasingly advised. Our study was launched to assess prospectively the clinical usefulness of systematic imatinib TDM in CML patients. This preliminary analysis addresses the appropriateness of the dosage adjustment approach applied in this study, which targets the recommended trough level and allows an interval of 4-24 h after last drug intake for blood sampling. Methods: Blood samples from the first 15 patients undergoing 1st TDM were obtained 1.5-25 h after last dose. Imatinib plasma levels were measured by LC-MS/MS and the concentrations were extrapolated to trough based on a Bayesian approach using a population pharmacokinetic model. Trough levels were predicted to differ significantly from the target in 12 patients (10 <750 μg/L; 2 >1500 μg/L along with poor tolerance) and individual dose adjustments were proposed. 8 patients underwent a 2nd TDM cycle. Trough levels of 1st and 2nd TDM were compared, the sample drawn 1.5 h after last dose (during distribution phase) was excluded from the analysis. Results: Individual dose adjustments were applied in 6 patients. Observed concentrations extrapolated to trough ranged from 360 to 1832 μg/L (median 725; mean 810, CV 52%) on 1st TDM and from 720 to 1187 μg/L (median 950; mean 940, CV 18%) on 2nd TDM cycle. Conclusions: These preliminary results suggest that TDM of imatinib using a Bayesian interpretation is able to target the recommended trough level of 1000 μg/L and to reduce the considerable differences in trough level exposure between patients (with CV decreasing from 52% to 18%). While this may simplify blood collection in daily practice, as samples do not have to be drawn exactly at trough, the largest possible interval to last drug intake yet remains preferable to avoid sampling during distribution phase leading to biased extrapolation. This encourages the evaluation of the clinical benefit of a routine TDM intervention in CML patients, which the randomized Swiss I-COME trial aims to.

Quantification of glucuronidated and sulfated steroids in human urine by ultra-high pressure liquid chromatography quadrupole time-of-flight mass spectrometry.

The urinary steroid profile is constituted by anabolic androgenic steroids, including testosterone and its relatives, that are extensively metabolized into phase II sulfated or glucuronidated steroids. The use of liquid chromatography coupled to mass spectrometry (LC-MS) is an issue for the direct analysis of conjugated steroids, which can be used as urinary markers of exogenous steroid administration in doping analysis, without hydrolysis of the conjugated moiety. In this study, a sensitive and selective ultra high-pressure liquid chromatography coupled to quadrupole time-of-flight mass spectrometer (UHPLC-QTOF-MS) method was developed to quantify major urinary metabolites simultaneously after testosterone intake. The sample preparation of the urine (1 mL) was performed by solid-phase extraction on Oasis HLB sorbent using a 96-well plate format. The conjugated steroids were analyzed by UHPLC-QTOF-MS(E) with a single-gradient elution of 36 min (including re-equilibration time) in the negative electrospray ionization mode. MS(E) analysis involved parallel alternating acquisitions of both low- and high-collision energy functions. The method was validated and applied to samples collected from a clinical study performed with a group of healthy human volunteers who had taken testosterone, which were compared with samples from a placebo group. Quantitative results were also compared to GC-MS and LC-MS/MS measurements, and the correlations between data were found appropriate. The acquisition of full mass spectra over the entire mass range with QTOF mass analyzers gives promise of the opportunity to extend the steroid profile to a higher number of conjugated steroids.

In vitro biotransformation of imatinib by the tumor expressed CYP1A1 and CYP1B1.

The main objective of the study was to examine the biotransformation of the anticancer drug imatinib in target cells by incubating it with oxidoreductases expressed in tumor cells. The second objective was to obtain an in silico prediction of the potential activity of imatinib metabolites. An in vitro enzyme kinetic study was performed with cDNA expressed human oxidoreductases and LC-MS/MS analysis. The kinetic parameters (Km and Vmax) were determined for six metabolites. A molecular modeling approach was used to dock these metabolites to the target Abl or Bcr-Abl kinases. CYP3A4 isozyme showed the broadest metabolic capacity, whereas CYP1A1, CYP1B1 and FMO3 isozymes biotransformed imatinib with a high intrinsic clearance. The predicted binding modes for the metabolites to Abl were comparable to that of the parent drug, suggesting potential activity. These findings indicate that CYP1A1 and CYP1B1, which are known to be overexpressed in a wide range of tumors, are involved in the biotransformation of imatinib. They could play a role in imatinib disposition in the targeted stem, progenitor and differentiated cancer cells, with a possible contribution of the metabolites toward the activity of the drug.

Alterações de atributos químicos e biológicos de solo e rendimento de milho e soja pela utilização de resíduos de curtume e carbonífero

Resíduos gerados por curtumes e pela exploração de carvão mineral são produtos potencialmente poluentes em várias regiões do Brasil, principalmente nos estados do Rio Grande do Sul e de Santa Catarina. O presente trabalho foi realizado no campo com o objetivo de avaliar o efeito da adição de resíduos de curtume e rejeito carbonífero sobre o solo e sobre as plantas cultivadas em um Argissolo Vermelho distrófico típico da Estação Experimental Agronômica da UFRGS, localizada no município de Eldorado do Sul (RS). O experimento foi realizado no ano agrícola de 1996/97, em parcelas de 70 m², cultivando-se soja (Glycine max L. Merrill) e milho (Zea mays L.). Tratamentos com a adição de 21,3 e 42,5 t ha-1 de lodo de curtume (LC) com adubação fosfatada e potássica na forma mineral foram comparados com o tratamento com adubação mineral completa (NPK) mais correção da acidez do solo e com a testemunha absoluta. Foram estudados também: (a) adição de resíduo carbonífero (106 t ha-1) mais adubação mineral; (b) resíduo carbonífero (106 t ha-1) mais lodo de curtume (21,3 t ha-1) com adubação fosfatada e potássica; (c) serragem cromada (29 t ha-1) mais adubação mineral (NPK), e (d) Cr mineral (125 kg ha-1) mais lodo de curtume (21,3 t ha-1) com adubação fosfatada e potássica. Foram avaliadas as alterações químicas e biológicas do solo, bem como os efeitos da aplicação dos resíduos sobre o rendimento e absorção de metais pelas plantas. A adição de LC aumentou o valor de pH e o teor de Ca trocável do solo. Não foram constatadas alterações nas concentrações dos metais Cu, Cd, Pb e Ni no solo, aumentando, entretanto, significativamente os teores de Cr. A atividade microbiana avaliada pela produção de CO2 foi estimulada pela adição dos resíduos, mas a população de bactérias, fungos e ctinomicetos não foi afetada. A adição de LC propiciou rendimentos de soja e milho semelhantes aos com adição de fertilizante nitrogenado. A adição de serragem cromada não alterou o rendimento das culturas, enquanto o rendimento de grãos de milho aumentou com a adição de resíduo carbonífero.

Eficiência de absorção, translocação e uso de cálcio, magnésio e enxofre por mudas enxertadas de Coffea arabica

O estudo da eficiência nutricional de plantas enxertadas de cafeeiro é importante para a seleção de combinações enxerto/porta-enxerto, visando ao desenvolvimento e produção máximos. O presente trabalho teve como objetivo avaliar, em cultivo hidropônico, a eficiência de absorção, translocação e uso de Ca, Mg e S por mudas enxertadas de Coffea arabica L., influenciada pelo porta-enxerto. O experimento foi realizado em casa de vegetação por um período de 170 dias, em vasos que continham areia, como substrato, e solução nutritiva circulante. Utilizaram-se, como enxerto, quatro genótipos de Coffea arabica L.: as variedades Catuaí Vermelho IAC 15 e Oeiras MG 6851 e as linhagens H 419-10-3-1-5 e H 514-5-5-3, e, como porta-enxerto, três genótipos de C. canephora Pierre et Froenher: Apoatã LC 2258, Conillon Muriaé-1, Robustão Capixaba (EMCAPA 8141) e um genótipo de Coffea arabica L.: Mundo Novo IAC 376-4, além da utilização de quatro pés-francos. Utilizou-se o delineamento experimental de blocos casualizados com quatro repetições. Os contrastes entre médias compararam as mudas de pé-franco com as associações enxerto/porta-enxerto. A eficiência de absorção, translocação e uso de Ca, Mg e S por mudas enxertadas de cafeeiro variou de acordo com a combinação enxerto/porta-enxerto. Somente a eficiência de translocação de Ca não foi alterada pela combinação enxerto/porta-enxerto. A linhagem H 514-5-5-3 foi beneficiada na eficiência de uso de Mg e produção de matéria seca pelos porta-enxertos Mundo Novo IAC 376-4 e Apoatã LC 2258, e na eficiência de uso de Ca e S apenas pelo Mundo Novo IAC 376-4.

Comparative protein profiling identifies elongation factor-1beta and tryparedoxin peroxidase as factors associated with metastasis in Leishmania guyanensis.

Parasites of the Leishmania Viannia subgenus are major causative agents of mucocutaneous leishmaniasis (MCL), a disease characterised by parasite dissemination (metastasis) from the original cutaneous lesion to form debilitating secondary lesions in the nasopharyngeal mucosa. We employed a protein profiling approach to identify potential metastasis factors in laboratory clones of L. (V.) guyanensis with stable phenotypes ranging from highly metastatic (M+) through infrequently metastatic (M+/M-) to non-metastatic (M-). Comparison of the soluble proteomes of promastigotes by two-dimensional electrophoresis revealed two abundant protein spots specifically associated with M+ and M+/M- clones (Met2 and Met3) and two others exclusively expressed in M- parasites (Met1 and Met4). The association between clinical disease phenotype and differential expression of Met1-Met4 was less clear in L. Viannia strains from mucosal (M+) or cutaneous (M-) lesions of patients. Identification of Met1-Met4 by biological mass spectrometry (LC-ES-MS/MS) and bioinformatics revealed that M+ and M- clones express distinct acidic and neutral isoforms of both elongation factor-1 subunit beta (EF-1beta) and cytosolic tryparedoxin peroxidase (TXNPx). This interchange of isoforms may relate to the mechanisms by which the activities of EF-1beta and TXNPx are modulated, and/or differential post-translational modification of the gene product(s). The multiple metabolic functions of EF-1 and TXNPx support the plausibility of their participation in parasite survival and persistence and thereby, metastatic disease. Both polypeptides are active in resistance to chemical and oxidant stress, providing a basis for further elucidation of the importance of antioxidant defence in the pathogenesis underlying MCL.