Multiple stages of detergent-erythrocyte membrane interaction-A spin label study

The various stages of the interaction between the detergent Triton X-100 (TTX-100) and membranes of whole red blood cells (RBC) were investigated in a broad range of detergent concentrations. The interaction was monitored by RBC hemolysis-assessed by release of intracellular hemoglobin (Hb) and inorganic phosphate- and by analysis of EPR spectra of a fatty acid spin probe intercalated in whole RBC suspensions, as well as pellets and supernatants obtained upon centrifugation of detergent-treated cells. Hemolysis finished at ca. 0.9 mM TTX-100. Spectral analysis and calculation of order parameters (S) indicated that a complex sequence of events takes place, and allowed the characterization of various structures formed in the different stages of detergent-membrane interaction. Upon reaching the end of cell lysis, essentially no pellet was detected, the remaining EPR signal being found almost entirely in the supernatants. Calculated order parameters revealed that whole RBC suspensions, pellets, and supernatants possessed a similar degree of molecular packing, which decreased to a small extent up to 2.5 mM detergent. Between 3.2 and 10 mM TTX-100, a steep decrease in S was observed for both whole RBC suspensions and supernatants. Above 10 mM detergent, S decreased in a less pronounced manner and the EPR spectra approached that of pure TTX-100 micelles. The data were interpreted in terms of the following events: at the lower detergent concentrations, an increase in membrane permeability occurs: the end of hemolysis coincides with the lack of pellet upon centrifugation. Up to 2.5 mM TTX-100 the supernatants consist of a (very likely) heterogeneous population of membrane fragments with molecular packing similar to that of whole cells. As the detergent concentration increases, mixed micelles are formed containing lipid and/or protein, approaching the packing found in pure TTX-100 micelles. This analysis is in agreement with the models proposed by Lasch (Biochim. Biophys Acta 1241 (1995) 269-292) and by Le Maire and coworkers (Biochim. Biophys. Acta 1508 (2000) 86-111). (C) 2010 Elsevier B.V. All rights reserved.

Polymethine cyanine dyes in beta-cyclodextrin solution: multiple equilibria and chemical oxidation

Absorption and fluorescence spectroscopy, electrochemical techniques, and semiempirical calculations were employed to characterize the multiple complexation equilibria between two polymethine cyanine dyes (IR-786 and Indocyanine green-ICG, 5) and beta-cyclodextrin (beta-CD, L), as well as the chemical reactivity of the complexed and uncomplexed species against the oxidizing agents hypochlorite (HC) and hydrogen peroxide (HP). IR-786 dimerization is favored with the increase in beta-CD concentration in the form of (SL)(2) complexes. In the case of ICG, free dimers (D) and SL complexes are favored. Both IR-786 and ICG react and discolor in the presence of HC and HP. For IR-786, the reaction with HP and HC proceeds with observed rate constants of 10(-3) and 0.28 s(-1) and second-order rate constants (k(2)) of similar to 10(-3) and 10(4) M(-1) s(-1), respectively. The intermediate species observed in the bleaching reactions of IR-786 and ICG were shown, by cyclic voltammetry and VIS absorption, to result from one electron oxidation. IR-786 complexed with beta-CD is protected against bleaching in the presence of HP and HC by factors of 20 and 4, respectively. This protection was not observed in ICG complexes. Superdelocalizability profile of both dyes and frontier orbital analysis indicates that beta-CD does not protect ICG from oxidation by HP or HC, whereas the 2:2 IR-786/beta-Cd complex is able to avoid the oxidation of IR-786. We concluded that the decrease in the chemical reactivity of the dyes against oxidant agents in the presence of beta-CD is due to the formation of (SL)(2) complexes. Copyright (C) 2010 John Wiley & Sons, Ltd.

Structure-activity relationships of hypervalent organochalcogenanes as inhibitors of cysteine cathepsins V and S

A new series of organotelluranes were synthesized and investigated, and the structure-activity relationships in cysteine proteases inhibition were determinated. It was possible to identify the relevance of structural components linked to the reactivity of these compounds as inhibitors. For example, dibromo-organotelluranes showed to be more reactive than dichloro-organotelluranes towards cysteine cathepsins V and S. Besides, no remarkable enantio-selectivity was verified. In general the achiral organotelluranes were more reactive than the chiral congeners against cysteine cathepsins V and S. A reactivity order for organochalcogenanes and cysteine cathepsins was proposed after the comparison of the inhibitory potencies of organotelluranes with the related organoselenanes. (C) 2011 Elsevier Ltd. All rights reserved.

Multiple large shareholders in corporate governance

Large shareholders of firms with majority bIocks are often at the heIm of their companies and do not necessarily have the same interests as minority shareholders. We show that bargaining problems Ied by the presence of muItipIe controlling shareholders protect minority shareholders. The same bargaining problems, however, prevent efficient decisions. By solving this trade-off we find that i) muItipIe controlling shareholders should be present in firms with Iarge costs of diIuting minority shareholders and in firms with Iarge financing requirements, ii) an optimal ownership structure requires the presence of a dass of shareholders - the minority shareholders - with no control over corporate decisions. Evidence on the ownership structure of dose corporations in the V.S. is consistent with our model.

Leadership of portuguese community schools in the East Coast of the United States and the public policies of education: a multiple case studies

The Portuguese community schools of the United States located in the areas of larger Portuguese population concentration are social organizations that come materializing throughout decades the designs of the educative policies of the Portuguese government in relation to the expansion and preservation of the language, the culture and the history of Portugal. These designs of the educative policies are enrolled in the Constitution of the Republic (1976), in the Basic Law of Educative System (1986) and, over all, in the successive legislative norms (Decree-laws and ordinances) of the successive governments. Portuguese community schools in the United States are structuralized in analogous way to schools of the Portuguese geographic space. For this qualitative study (multiple case), four directors of Portuguese schools of the East Coast of the United States were interviewed; two schools are in the state of Rhode Island and the other two are in the state of Massachusetts. Also, it was administered the questionnaire on practices of leadership “Leadership Practices Inventory” (LPI) of Kouzes and Posner (2002) to collect additional data about practices of leadership on the directors of the schools. The LPI evaluates practices of leadership classifying them in five domains: (a) Model the way; (b) Inspire a shared vision; (c) Challenge the process; (d) Enable others to act; and, (e) Encourage the heart. Results of this qualitative research indicate that the Portuguese Government has not had an educative policy stimulant, coherent and consistent of support, incentive, maintenance and diffusion of the Portuguese language and culture and the directors of the studied schools they have a proactive and serving leadership style in conducting the management of Portuguese community schools. The five practices of leadership are highly practiced by the directors of the studied schools above all the practices “Enable others to act” and “Encourage the heart”.

Effect of sequence of insemination after simultaneous thawing of multiple semen straws on conception rate to timed AI in suckled multiparous Nelore cows

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Modelling energy utilisation in broiler breeder hens

1. The objective of this study was to determine a metabolisable energy ( ME) requirement model for broiler breeder hens. The influence of temperature on ME requirements for maintenance was determined in experiments conducted in three environmental rooms with temperatures kept constant at 13, 21 and 30 degrees C using a comparative slaughter technique. The energy requirements for weight gain were determined based upon body energy content and efficiency of energy utilisation for weight gain. The energy requirements for egg production were determined on the basis of egg energy content and efficiency of energy deposition in the eggs.2. The following model was developed using these results: ME = kgW(0.75)(806.53 - 26.45T + 0.50T(2)) + 31.90G + 10.04EM, where kgW(0.75) is body weight (kg) raised to the power 0.75, T is temperature (degrees C), G is weight gain (g) and EM is egg mass (g).3. A feeding trial was conducted using 400 Hubbard Hi-Yield broiler breeder hens and 40 Peterson males from 31 to 46 weeks of age in order to compare use of the model with a recommended feeding programme for this strain of bird. The application of the model in breeder hens provided good productive and reproductive performance and better results in feed and energy conversion than in hens fed according to strain recommendation. In conclusion, the model evaluated predicted an ME intake which matched breeder hens' requirements.

Random regressions models to describe the genetic variation of milk yield over multiple parities in Buffaloes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Splice variants of the human ZC3H14 gene generate multiple isoforms of a zinc finger polyadenosine RNA binding protein

The human ZC3H14 gene encodes an evolutionarily conserved Cys(3)His zinc finger protein that binds specifically to polyadenosine RNA and is thus postulated to modulate post-transcriptional gene expression. Expressed sequence tag (EST) data predicts multiple splice variants of both human and mouse ZC3H14. Analysis of ZC3H14 expression in both human cell lines and mouse tissues confirms the presence of multiple alternatively spliced transcripts. Although all of these transcripts encode protein isoforms that contain the conserved C-terminal zinc finger domain, suggesting that they could all bind to polyadenosine RNA, they differ in other functionally important domains. Most of the alternative transcripts encode closely related proteins (termed isoforms 1, 2. 3, and 3short) that differ primarily in the inclusion of three small exons, 9, 10, and 11, resulting in predicted protein isoforms ranging from 82 to 64 kDa. Each of these closely related isoforms contains predicted classical nuclear localization signals (cNLS) within exons 7 and 11. Consistent with the presence of these putative nuclear targeting signals, these ZC3H14 isoforms are all localized to the nucleus. In contrast, an additional transcript encodes a smaller protein (34 kDa) with an alternative first exon (isoform, 4). Consistent with the absence of the predicted cNLS motifs located in exons 7 and 11, ZC3H14 isoform 4 is localized to the cytoplasm. Both EST data and experimental data suggest that this variant is enriched in testes and brain. Using an antibody that detects endogenous ZC3H14 isoforms 1-3 reveals localization of these isoforms to nuclear speckles. These speckles co-localize with the splicing factor, SC35, suggesting a role for nuclear ZC3H14 in mRNA processing. Taken together, these results demonstrate that multiple transcripts encoding several ZC3H14 isoforms exist in vivo. Both nuclear and cytoplasmic ZC3H14 isoforms could have distinct effects on gene expression mediated by the common Cys(3)His zinc finger polyadenosine RNA binding domain. (C) 2009 Elsevier B.V. All rights reserved.

High-energy transitions of shallow magnetodonors in a GaAs/Al0.3Ga0.7As multiple quantum well

The high-energy states of a shallow donor in a GaAs/Ga0.7Al0.3As multiple-quantum-well structure subjected to a magnetic field in the growth direction are studied both theoretically and experimentally. Effects due to higher confinement subbands as well as due to the electron-phonon interaction are investigated. We show that most of the peaks in the infrared photoconductivity spectrum are due to direct transitions from the ground state to the m = +/-1 magnetodonor states associated with the first subband, but transitions to the m = +/-1 states of the third subband are also apparent. The remaining photoconductivity peaks are explained by phonon-assisted impurity transitions.

Ultrasonic densitometer using a multiple reflection technique

This work presents recent improvements in a density measurement cell with a double-element transducer that can eliminate diffraction effects. A new mechanical design combined with the use of more appropriate materials has resulted in better parallelism between interfaces, more robust assembly, and chemical resistance. A novel method of signal processing, named energy method, is introduced to obtain the reflection coefficient, reducing sensitivity to noise and improving accuracy. The measurement cell operation is verified both theoretically, using an acoustic wave propagation model, and experimentally, using homogeneous liquids with different densities. The accuracy in the density measurement is 0.2% when compared with the measurements made with a pycnometer.

Absence of DNA damage in multiple organs (blood, liver, kidney, thyroid gland and urinary bladder) after acute fluoride exposure in rats

Fluoride has been widely used in dentistry as a caries prophylactic agent. However, there has been some speculation that excess fluoride could cause an impact on genome integrity. In the current study, the potential DNA damage associated with exposure to fluoride was assessed in cells of blood, liver, kidney, thyroid gland and urinary bladder by the single cell gel (comet) assay. Male Wistar rats aging 75 days were distributed into seven groups: Groups 1 (control), 2, 3, 4, 5, 6 and 7 received 0 (deionized water), 10, 20, 40, 60, 80 and 100 mgF/Kg body weight from sodium fluoride (NaF), respectively, by gastrogavage. These groups were killed at 2 h after the administration of the fluoride doses. The level of DNA strand breaks did not increase in all organs evaluated and at all doses of NaF tested, as depicted by the mean tail moment. Taken together, our results suggest that oral exposure to NaF did not result in systemic genotoxic effect in multiple organs related to fluoride toxicity. Since DNA damage is an important step in events leading to carcinogenesis, this study represents a relevant contribution to the correct evaluation of the potential health risk associated with chemical exposure.