Stress, coping and personal strengths and difficulties in internationally adopted children in spain

This study analyses the types of coping strategies used by internationally adopted children, and explores the relation between these strategies and personal strengths and difficulties. The Kidcope checklist (Spirito, Stark, & Williams, 1998) and the Strengths and Difficulties Questionnaire (SDQ; Goodman, 1997) were administered to a sample of 35 Spanish adoptees (25.7% boys and 74.3% girls, aged 8-12 years) and their parents. Self-reported problems were categorised and their relation with coping strategies and psychological adjustment was explored. Results indicated that adopted children report problems of interpersonal nature. The content of the problems mainly refers to relationships and health, illness, or accidents. Parents reported that children were generally well-adjusted and they had no problems outside the normal range. International adoptees used mainly control-oriented coping strategies. Escape-oriented coping was linked to parents' ratings of total difficulties, with self-criticism accounting for the highest percentage of the variance.

Role of Munc18c in SNARE-mediated exocytosis

Background: The SNARE (Soluble N-ethylmaleimide-sensitive factor Attachment protein Receptors) and SM (Sec1/Munc18) family of proteins form the core machinery that drives the fusion of vesicles in different membrane trafficking steps. They are highly conserved, implying a similar mode of binding and function. In vertebrates, Munc18a is essential for neuronal exocytosis. It binds to its partner syntaxin1a (Syx1a) at both its N-peptide and closed conformation, and thereby inhibits SNARE complex formation in vitro. By contrast, its close homolog Munc18c is thought to interact with only the N-peptide of its partner Syx4. Moreover, different effects of Munc18c on SNARE complex formation have been reported, suggesting that the two Munc18/Syx pairs act differently. Objective: The aim of the present study was to investigate whether the mechanism of action of Munc18c indeed deviates from that of Munc18a by using sensitive biochemical and biophysical methods. Results: I found that Munc18c does have a similar binding mode as Munc18a and interacts tightly with Syx4 at both the N-peptide and closed conformation. Moreover, I established, through a novel assay, that Munc18c inhibits SNARE complex assembly, with both the binding sites contributing to inhibition, similar to Munc18a. However, there were several subtle differences between the two Munc18/Syx pairs. Munc18a exerted stronger inhibition than Munc18c. Also their respective Syx partners were found to differ in the rate of binding to SNAP25, suggesting that the equilibrium of their open and closed conformations is different. Moreover, Munc18a was found to interact with Syx 1, 2, 3 but not 4, while Munc18c bound to Syx 2, 4 and 1 but not 3. By comparing the kinetics of interaction of Syx with either Munc18 or SNAP25, I found that the block of SNARE complex assembly by Munc18 is effective on a shorter time scale, but SNAP25 eventually binds to Syx resulting in SNARE complex formation. Nevertheless, these findings do not explain how Syx can escape the tight grip of Munc18, suggesting that other proteins or mechanisms are needed for this step. I also discovered that Munc18 is able to bind on the surface of the SNARE core complex; however, this observation needs to be tested more rigorously. Conclusion: Munc18c was found to be similar to Munc18a in its mode of binding to Syx and inhibition of SNARE complex assembly. However, differences in kinetics and interaction specificities were observed between the different Munc18/Syx pairs. -- Contexte : Les familles des protéines SNARE (Soluble N-ethylmaleimide-sensitive factor At- tachment protein Receptors) et SM (Sec1/Munc18) forment le coeur de la machinerie chargée de la fusion vésiculaire au cours des différentes étapes du trafic intracellulaire. Elles sont très conservées, suggérant un mode d'interaction et des fonctions semblables. Chez les Verté- brés, Munc18a est essentielle à l'exocytose neuronale. Elle se lie à sa partenaire d'interaction syntaxin1a (Syx1a) à la fois via un peptide N-terminal et la conformation fermée de celle-ci, inhibant ainsi la formation du complexe SNARE in vitro. Son homologue proche Munc18c au contraire, est supposée interagir seulement avec le peptide N-terminal de sa partenaire Syx4. En outre, différents effets de Munc18c sur la formation du complexe SNARE ont été décrits, suggérant que les deux paires Munc18/Syx fonctionnent différemment. Objectif : Le but de cette étude est de tester si les mécanismes de fonctionnement de Munc18c diffèrent vraiment de ceux de Munc18a par le biais de méthodes biochimiques et biophysiques très précises. Résultats : J'ai pu démontrer que Munc18c se comporte en effet de façon semblable à Munc18a, et interagit étroitement avec Syx4 à ses deux sites de liaison. J'ai pu de surcroît montrer par une nouvelle méthode que Munc18c inhibe l'assemblage du complexe SNARE en impliquant ces deux sites de liaison, comme le fait Munc18a. il existe cependant de subtiles différences entre les deux paires Munc18/Syx : Munc18a exerce une inhibition plus forte que Munc18c ; leurs Syx partenaires diffèrent également dans leur degré de liaison à SNAP25, ce qui suggère un équilibre different de leurs conformations ouverte et fermée. De plus, Munc18a interagit avec Syx 1, 2 et 3 mais pas Syx 4, alors que Munc18c se lie à Syx 2, 4 et 1 mais pas Syx 3. En comparant les cinétiques d'interaction de Syx avec Munc18 ou SNAP25, j'ai découvert que le blocage par Munc18 de l'assemblage du complexe SNARE est effectif de façon brève, bien que SNAP25 finisse par se lier à Syx et aboutir ainsi à la formation du complexe SNARE. Ces découvertes n'expliquent cependant pas comment Syx parvient à échapper à la solide emprise de Munc18, et suggèrent ainsi l'intervention nécessaire d'autres protéines ou mécanismes à cette étape. J'ai également découvert que Munc18 peut se lier à la surface de la partie centrale du complexe SNARE - cette observation reste à être testée de façon plus stringente. Conclusion : Il a pu être établi que Munc18c est semblable à Munc18a quant à son mode de liaison à Syx et d'inhibition de l'assemblage du complexe SNARE. Des différences de cinétique et de spécificité d'interaction entre les diverses paires Munc18/Syx ont cependant été identifiées.

Stabbing simulations and DNA transfer

Technical developments have made it possible to analyze very low amounts of DNA. This has many advantages, but the drawback of this technological progress is that interpretation of the results becomes increasingly complex: the number of mixed DNA profiles increased relatively to single source DNA profiles and stochastic effects in the DNA profile, such as drop-in and drop-out, are more frequently observed. Moreover, the relevance of low template DNA material regarding the activities alleged is not as straightforward as it was a few years ago, when for example large quantities of blood were recovered. The possibility of secondary and tertiary transfer is now becoming an issue. The purpose of this research is twofold: first, to study the transfer of DNA from the handler and secondly, to observe if handlers would transfer DNA from persons closely connected to them. We chose to mimic cases where the offender would attack a person with a knife. As a first approach, we envisaged that the defense would not give an alternative explanation for the origin of the DNA. In our transfer experiments (4 donors, 16 experiments each, 64 traces), 3% of the traces were single DNA profiles. Most of the time, the DNA profile of the person handling the knife was present as the major profile: in 83% of the traces the major contributor profile corresponded to the stabber's DNA profile (in single stains and mixtures). Mixture with no clear major/minor fraction (12%) were observed. 5% of the traces were considered of insufficient quality (more than 3 contributors, presence of a few minor peaks). In that case, we considered that the stabber's DNA was absent. In our experiments, no traces allowed excluding the stabber, however it must be noted that precautions were taken to minimize background DNA as knives were cleaned before the experiments. DNA profiles of the stabber's colleagues were not observed. We hope that this study will allow for a better understanding of the transfer mechanism and of how to assess and describe results given activity level propositions. In this preliminary research, we have focused on the transfer of DNA on the hand of the person. Besides, more research is needed to assign the probability of the results given an alternative activity proposed by the defense, for instance when the source of the DNA is not contested, but that the activities are.

Thermal Degradation of Small Molecules: A Global Metabolomic Investigation.

Thermal processes are widely used in small molecule chemical analysis and metabolomics for derivatization, vaporization, chromatography, and ionization, especially in gas chromatography mass spectrometry (GC/MS). In this study the effect of heating was examined on a set of 64 small molecule standards and, separately, on human plasma metabolite extracts. The samples, either derivatized or underivatized, were heated at three different temperatures (60, 100, and 250 °C) at different exposure times (30 s, 60 s, and 300 s). All the samples were analyzed by liquid chromatography coupled to electrospray ionization mass spectrometry (LC/MS) and the data processed by XCMS Online ( xcmsonline.scripps.edu ). The results showed that heating at an elevated temperature of 100 °C had an appreciable effect on both the underivatized and derivatized molecules, and heating at 250 °C created substantial changes in the profile. For example, over 40% of the molecular peaks were altered in the plasma metabolite analysis after heating (250 °C, 300s) with a significant formation of degradation and transformation products. The analysis of 64 small molecule standards validated the temperature-induced changes observed on the plasma metabolites, where most of the small molecules degraded at elevated temperatures even after minimal exposure times (30 s). For example, tri- and diorganophosphates (e.g., adenosine triphosphate and adenosine diphosphate) were readily degraded into a mono-organophosphate (e.g., adenosine monophosphate) during heating. Nucleosides and nucleotides (e.g., inosine and inosine monophosphate) were also found to be transformed into purine derivatives (e.g., hypoxanthine). A newly formed transformation product, oleoyl ethyl amide, was identified in both the underivatized and derivatized forms of the plasma extracts and small molecule standard mixture, and was likely generated from oleic acid. Overall these analyses show that small molecules and metabolites undergo significant time-sensitive alterations when exposed to elevated temperatures, especially those conditions that mimic sample preparation and analysis in GC/MS experiments.

Direct Interrogation of Viral Peptides Presented by the Class I HLA of HIV-Infected T Cells

Identification of CD8+ cytotoxic T lymphocyte (CTL) epitopes has traditionally relied upon testing of overlapping peptide libraries for their reactivity with T cells in vitro. Here, we pursued deep ligand sequencing (DLS) as an alternative method of directly identifying those ligands that are epitopes presented to CTLs by the class I human leukocyte antigens (HLA) of infected cells. Soluble class I HLA-A*11:01 (sHLA) was gathered from HIV-1 NL4-3-infected human CD4+ SUP-T1 cells. HLA-A*11:01 harvested from infected cells was immunoaffinity purified and acid boiled to release heavy and light chains from peptide ligands that were then recovered by size-exclusion filtration. The ligands were first fractionated by high-pH high-pressure liquid chromatography and then subjected to separation by nano-liquid chromatography (nano-LC)–mass spectrometry (MS) at low pH. Approximately 10 million ions were selected for sequencing by tandem mass spectrometry (MS/MS). HLA-A*11:01 ligand sequences were determined with PEAKS software and confirmed by comparison to spectra generated from synthetic peptides. DLS identified 42 viral ligands presented by HLA-A*11:01, and 37 of these were previously undetected. These data demonstrate that (i) HIV-1 Gag and Nef are extensively sampled, (ii) ligand length variants are prevalent, particularly within Gag and Nef hot spots where ligand sequences overlap, (iii) noncanonical ligands are T cell reactive, and (iv) HIV-1 ligands are derived from de novo synthesis rather than endocytic sampling. Next-generation immunotherapies must factor these nascent HIV-1 ligand length variants and the finding that CTL-reactive epitopes may be absent during infection of CD4+ T cells into strategies designed to enhance T cell immunity.

Predator or prey? Chlamydophila abortus infections of a free-living amoebae, Acanthamoeba castellani 9GU.

Limited evidence exists to suggest that the ability to invade and escape protozoan host cell bactericidal activity extends to members of the Chlamydiaceae, intracellular pathogens of humans and animals and evolutionary descendants of amoeba-resisting Chlamydia-like organisms. PCR and microscopic analyses of Chlamydophila abortus infections of Acanthamoeba castellani revealed uptake of this chlamydial pathogen but, unlike the well-described inhabitant of A. castellani, Parachlamydia acanthamoebae, Cp. abortus did not appear to propagate and is likely digested by its amoebal host. These data raise doubts about the ability of free-living amoebae to serve as hosts and vectors of pathogenic members of the Chlamydiaceae but reveal opportunities, via comparative genomics, to understand virulence mechanisms used by Chlamydia-like organisms to avoid amoebal digestion.

Immediate ecotoxicological effects of short-lived oil spills on marine biota.

Marine environments are frequently exposed to oil spills as a result of transportation, oil drilling or fuel usage. Whereas large oil spills and their effects have been widely documented, more common and recurrent small spills typically escape attention. To fill this important gap in the assessment of oil-spill effects, we performed two independent supervised full sea releases of 5 m(3) of crude oil, complemented by on-board mesocosm studies and sampling of accidentally encountered slicks. Using rapid on-board biological assays, we detect high bioavailability and toxicity of dissolved and dispersed oil within 24 h after the spills, occurring fairly deep (8 m) below the slicks. Selective decline of marine plankton is observed, equally relevant for early stages of larger spills. Our results demonstrate that, contrary to common thinking, even small spills have immediate adverse biological effects and their recurrent nature is likely to affect marine ecosystem functioning.

Excitação eletrônica das moléculas de metacrilato de metila e estireno na região do ultravioleta de vácuo

Angle-resolved electron energy-loss spectra have been measured for the methyl methacrylate (MMA) and styrene molecules in the 0 - 50 eV energy range. The spectra have been obtained at 1 keV incident energy, with an energy resolution of 0.8 eV and covering an angular range of 2.0 to 7.0 degrees. Within our knowledge, this is the first gas-phase excitation spectrum for MMA and styrene in this energy range. The spectra of MMA at small scattering angles are dominated by an intense peak at 6.7 eV followed by a broad band centered at about 16 eV. In the case of styrene, six bands can be observed in the spectra. Based on the angular behaviour of the excitation spectra of these molecules, the low-lying peaks observed are considered to be associated predominantly with dipole-allowed processes. In both cases, new bands can be observed for excitation energies greater than 20 eV. This could be associated with dipole-forbidden transitions to shake-up and doubly-excited states.

Identificação dos principais componentes do fungicida guazatine através da cromatografia a gás e cromatografia a gás acoplada com espectrometria de massas

Hexafluoroacetylaceton derivatives of some synthesized components of the guazatine fungicide were prepared, and their separation and identification performed through gas chromatography and gas chromatography-mass spectrometry. The chromatogram obtained from hexafluoroacetylacetone derivative of the commercial guazatine presented 14 major peaks, and 9 of them were identified through both techniques mentioned above.

Imaging polarimetry of the fogbow: polarization characteristics of white rainbows measured in the high Arctic

The knowledge on the optics of fogbows is scarce, and their polarization characteristics have never been measured to our knowledge. To fill this gap we measured the polarization features of 16 fogbows during the Beringia 2005 Arctic polar research expedition by imaging polarimetry in the red, green and blue spectral ranges. We present here the first polarization patterns of the fogbow. In the patterns of the degree of linear polarization p, fogbows and their supernumerary bows are best visible in the red spectral range due to the least dilution of fogbow light by light scattered in air. In the patterns of the angle of polarization α fogbows are practically not discernible because their α-pattern is the same as that of the sky: the direction of polarization is perpendicular to the plane of scattering and is parallel to the arc of the bow, independently of the wavelength. Fogbows and their supernumeraries were best seen in the patterns of the polarized radiance. In these patterns the angular distance δ between the peaks of the primary and the first supernumerary and the angular width σ of the primary bow were determined along different radii from the center of the bow. δ ranged between 6.08° and 13.41° , while σ changed from 5.25° to 19.47° . Certain fogbows were relatively homogeneous, meaning small variations of δ and σ along their bows. Other fogbows were heterogeneous, possessing quite variable δ- and σ-values along their bows. This variability could be a consequence of the characteristics of the high Arctic with open waters within the ice shield resulting in the spatiotemporal change of the droplet size within the fog

Fotodegradação de polímeros clorados irradiados com a luz síncrotron - I. Emissão de gases

Chlorinated polymers (PVC, PVDC and E-CTFE) were irradiated with white light produced at the Brazilian Synchrotron Light Source (LNLS). The emitted gases were analyzed by mass spectrometry. The spectra were dominated by peaks related to hydrochloric acid, HCl, and chlorine (35Cl). The measured HCl intensity is used to evaluate the sensitivity of the polymers over a broad energy range. PVDC showed the greatest light sensitivity as compared to PVC and E-CTFE.

Estudo termogravimétrico e calorimétrico da algaroba

Humidity and ash content of powder and bran of algaroba pods are important quality parameters for biological control during storage. The studies of humidity and ash content were carried out conventional and thermogravimetric methods. Different values were obtained by the various methods. The thermogravimetric method was faster and required less sample for the humidity and ash analyses of the powder and bran of algaroba pods. The differential scanning calorimetry curves of the powder and bran of algaroba pods, dryed at 55 ºC, showed two peaks, one corresponding to the gelatinization of starch and the other to the vaporization of water. The samples dryed at 65, 75, 85, 95 and 105 ºC showed one peak, corresponding to the vaporization of water.

Estudo comparativo sobre filtragem de sinais instrumentais usando transformadas de Fourier e Wavelet

A comparative study of the Fourier (FT) and the wavelet transforms (WT) for instrumental signal denoising is presented. The basic principles of wavelet theory are described in a succinct and simplified manner. For illustration, FT and WT are used to filter UV-VIS and plasma emission spectra using MATLAB software for computation. Results show that FT and WT filters are comparable when the signal does not display sharp peaks (UV-VIS spectra), but the WT yields a better filtering when the filling factor of the signal is small (plasma spectra), since it causes low peak distortion.

Determinação espectrofotométrica de dipirona em produtos farmacêuticos por injeção em fluxo pela geração de íons triiodeto

A flow injection spectrophotometric procedure was developed for the determination of metamizol in pharmaceutical formulations. The system is based on the reaction between metamizol and triiodide generated in the system by mixing iodate and iodide-starch solutions. The absorbance of triiodide-starch complex giving a steady-state baseline value which was monitored at 580 nm. The inverse peaks caused by metamizol samples were measured and there was a direct relationship between absorbance decreasing and metamizol concentration from 1.4 x 10-4 to 7.0 x 10-4 mol L-1. The RSD was 0.45 % for a metamizol solution 4.2 x 10-4 mol L-1 (n = 10) with a detection limit (three-fold blank standard deviation/slope) of 6.0 x 10-5 mol L-1 The feasibility of the system was demonstrated for the determination of metamizol in commercial samples with sixty results obtained per hour. The results obtained for metamizol in pharmaceutical formulations using the proposed flow procedure and those obtained using an iodimetric procedure are in agreement at the 95% confidence level and within an acceptable range of error.

Methods to test gas tightness of packages

One of the primary goals for food packages is to protect food against harmful environment, especially oxygen and moisture. The gas transmission rate is the total gas transport through the package, both by permeation through the package material and by leakage through pinholes and cracks. The shelf life of a product can be extended, if the food is stored in a gas tight package. Thus there is a need to test gas tightness of packages. There are several tightness testing methods, and they can be broadly divided into destructive and nondestructive methods. One of the most sensitive methods to detect leaks is by using a non destructive tracer gas technique. Carbon dioxide, helium and hydrogen are the most commonly used tracer gases. Hydrogen is the lightest and the smallest of all gases, which allows it to escape rapidly from the leak areas. The low background concentration of H2 in air (0.5 ppm) enables sensitive leak detection. With a hydrogen leak detector it is also possible to locate leaks. That is not possible with many other tightness testing methods. The experimental work has been focused on investigating the factors which affect the measurement results with the H2leak detector. Also reasons for false results were searched to avoid them in upcoming measurements. From the results of these experiments, the appropriate measurement practice was created in order to have correct and repeatable results. The most important thing for good measurement results is to keep the probe of the detector tightly against the leak. Because of its high diffusion rate, the HZ concentration decreases quickly if holding the probe further away from the leak area and thus the measured H2 leaks would be incorrect and small leaks could be undetected. In the experimental part hydrogen, oxygen and water vapour transmissions through laser beam reference holes (diameters 1 100 μm) were also measured and compared. With the H2 leak detector it was possible to detect even a leakage through 1 μm (diameter) within a few seconds. Water vapour did not penetrate even the largest reference hole (100 μm), even at tropical conditions (38 °C, 90 % RH), whereas some O2 transmission occurred through the reference holes larger than 5 μm. Thus water vapour transmission does not have a significant effect on food deterioration, if the diameter of the leak is less than 100 μm, but small leaks (5 100 μm) are more harmful for the food products, which are sensitive to oxidation.