Crystal Structure of the Human Natural Killer Cell Activating Receptor KIR2DS2 (CD158j)

Killer cell Ig-like receptors (KIRs) regulate the function of human natural killer and T cell subsets. A feature of the KIR locus is the clustering of homologous genes encoding for inhibitory and activating KIR. Inhibitory and activating KIR differ for ligand specificities and/or affinities. In particular, we show here with KIR tetramers that activating KIR2DS2 does not bind HLA-Cw3 molecules recognized by inhibitory KIR2DL2, despite 99% extracellular amino acid identity. We also report the 2.3-Å structure of KIR2DS2, which reveals subtle displacements of two residues (Tyr45 and Gln71) involved in the interaction of KIR2DL2 with HLA-Cw3. These results show that KIR molecules cannot tolerate any variability in their three-dimensional structure without altering their MHC class I recognition capacities. Therefore, the mode of recognition used by KIR largely differs from the conformational changes that characterize T cell receptor or NKG2D interaction with their respective ligands

A new strategy for faster urinary biomarkers identification by Nano-LC-MALDI-TOF/TOF mass spectrometry

<p>Abstract</p> <p>Background</p> <p>LC-MALDI-TOF/TOF analysis is a potent tool in biomarkers discovery characterized by its high sensitivity and high throughput capacity. However, methods based on MALDI-TOF/TOF for biomarkers discovery still need optimization, in particular to reduce analysis time and to evaluate their reproducibility for peak intensities measurement. The aims of this methodological study were: (i) to optimize and critically evaluate each step of urine biomarker discovery method based on Nano-LC coupled off-line to MALDI-TOF/TOF, taking full advantage of the dual decoupling between Nano-LC, MS and MS/MS to reduce the overall analysis time; (ii) to evaluate the quantitative performance and reproducibility of nano-LC-MALDI analysis in biomarker discovery; and (iii) to evaluate the robustness of biomarkers selection.</p> <p>Results</p> <p>A pool of urine sample spiked at increasing concentrations with a mixture of standard peptides was used as a specimen for biological samples with or without biomarkers. Extraction and nano-LC-MS variabilities were estimated by analyzing in triplicates and hexaplicates, respectively. The stability of chromatographic fractions immobilised with MALDI matrix on MALDI plates was evaluated by successive MS acquisitions after different storage times at different temperatures.</p> <p>Low coefficient of variation (CV%: 10–22%) and high correlation (R²> 0.96) values were obtained for the quantification of the spiked peptides, allowing quantification of these peptides in the low fentomole range, correct group discrimination and selection of "specific" markers using principal component analysis. Excellent peptide integrity and stable signal intensity were found when MALDI plates were stored for periods of up to 2 months at +4°C. This allowed storage of MALDI plates between LC separation and MS acquisition (first decoupling), and between MS and MSMS acquisitions while the selection of inter-group discriminative ions is done (second decoupling). Finally the recording of MSMS spectra to obtain structural information was focused only on discriminative ions in order to minimize analysis time.</p> <p>Conclusion</p> <p>Contrary to other classical approaches with direct online coupling of chromatographic separation and on the flight MS and/or MSMS data acquisition for all detected analytes, our dual decoupling strategy allowed us to focus on the most discriminative analytes, giving us more time to acquire more replicates of the same urine samples thus increasing detection sensitivity and mass precision.</p

Efecto en las propiedades mecánicas de una resina pinífera modificada biodegradable, al utilizarla como compatibilizante o acoplante en formulaciones elastoméricas sin o con fibra de agave y hule de poli(estireno-butadieno). Un paso hacia la formulación

En este trabajo se valora la eficiencia de una resina de pino modificada biodegradable (A-30), al utilizarla como compatibilizante o acoplante para sustituir al resorcinol (que es tóxico y no es biodegradable), comparando las propiedades mecánicas (contra formulaciones preparadas usando el sistema resorcinol-hexametilentetramina (R-HMT) en: a) polímeros elastoméricos reticulados de estireno-butadieno (SBR), y: b) compósitos de SBR reforzados con fibra de agave de desecho. Para la comparación como compatibilizante, aquí se preparan formulaciones equivalentes con A-30 o R-HMT. Para la comparación de los compósitos equivalentes, se varía el agente de acoplamiento (R-HMT o A-30) y la cantidad de azufre como agente de entrecruzamiento (2,5 partes por cien de elastómero (phr), 7,5 phr o 15 phr). En las pruebas de tracción a temperatura ambiente, y el análisis mecanodinámico con barridos de temperatura (de - 70 °C a 40 °C) y frecuencia (de 0,4 Hz a 40 Hz), se obtuvieron incrementos en módulos (hasta un 74 % en módulo de Young) y en capacidad de deformación (hasta 260 %), para los materiales conteniendo el agente A-30, por encima de las formulaciones en las que se usó el resorcinol. El mejor desempeño como compatibilizante o acoplante del A-30, se reflejó también con claridad morfológicamente mediante microscopía electrónica de barrido, al mostrar mayor compacidad en la matriz formulada y mayor cercanía entre fibra y matriz en muestras fracturadas criogénicamente. Se ha demostrado aquí, que el A-30 puede sustituir al par R-HMT con éxito, ya que funciona mejor como agente compatibilizante o acoplante y es biodegradable. Este tipo de agentes tiene un gran potencial para contribuir al desarrollo de elastómeros y compósitos flexibles amigables con el medio ambiente

Understanding the National Student Survey: investigations in languages, linguistics and area studies

This report is a summary of interviews and focus groups with around 100 students and 50 members of academic staff in departments of languages, linguistics or area studies at nine universities in the UK. In recent years, concerns have been expressed about the ambiguity of some of the statements which students are asked to respond to in the National Student Survey (NSS). This project set out to get a better understanding of how students and staff understand the questions. The interviews and focus groups were carried out by members of academic staff at the nine institutions who each then wrote an individual report of their findings. This summary is designed to enable wider distribution of these findings without identifying individual staff, institutions `or departments

Valoración mecanotérmica de una resina biodegradable como agente de acoplamiento de materiales compuestos celulósicos/polímero hidrofóbico

En la búsqueda de plásticos reforzados con fibras que sean más amigables con el medio ambiente, aquí se presenta el primer estudio que evalúa la posibilidad de utilizar la brea natural de pino (en forma pura o maleinizada) como agente de acoplamiento (biodegradable). Polipropileno (matriz) y fibra de agave (Agave tequilana) de desecho a diferentes concentraciones (agente de refuerzo), fueron acoplados con cada uno de los agentes utilizados; su efecto en las propiedades mecánicas se comparó con el de un agente comercial de polipropileno modificado (Epolene E-43). Igualmente se prepararon, materiales compuestos sin agente de acoplamiento como referencia genérica. El desempeño mecanodinámico y mecanoestático de los materiales muestra claramente el incremento de propiedades mecánicas con los 3 agentes utilizados. La brea maleinizada mostró similitud o ligera superioridad sobre el agente comercial en el efecto logrado. La afectación en cristalinidad por la presencia de la fibra y el agente de acoplamiento correspondiente, fue evaluada mediante calorimetría diferencial de barrido. La absorción de agua como función del tiempo, permitió medir de forma indirecta el cambio logrado en la superficie de los materiales, y un análisis de FTIR, la valoración de la interacción fibra-polímero obtenida con el agente de acoplamiento. Tal interacción lograda con los agentes de acoplamiento, pudo ser además apreciada utilizando microscopía electrónica de barrido. Los resultados alcanzados marcan el camino para poder usar resinas naturales biodegradables como agentes de acoplamiento en el área de plásticos reforzados con fibras celulósicas. AbstractIn the search of useful environmentally friendly fiber reinforced plastics, this is the first study that evaluates the capability to use natural pine rosin (in pure or maleated glycerol ester form) as a biodegradable coupling agent. Polypropylene as polymer matrix and discarded agave fiber (Agave tequilana) as reinforcing agent at different concentrations, were coupled with each one of the two rosins above mentioned; a commercial maleated polypropylene (Epolene, E-43) agent was used to compare their effect. As generic reference, composites without coupling agent were also tested. Mechanodynamic and mechanostatic tests clearly show an increment in mechanical properties of the composites, using any of the 3 coupling agents. The results obtained with maleated rosin were similar or slightly better than the ones obtained with the commercial agent for composites with high fiber content. Fiber content and coupling agent effect on composites crystallinity, was evaluated by differential scanning calorimetry. In addition, water absorption as a function of time was followed to evaluate the effect of surface modification, and FTIR analysis allowed the observation of the fiber-polymer matrix interaction that was promoted with the coupling agents. The effect of such interaction obtained with the different coupling agents, was observed by scanning electron microscopy. The results show the feasibility to use the natural pine rosin in pure or modified form as biodegradable coupling agents

Thermal degradation kinetics of real-life reclaimed plastic solid waste (PSW) from an active landfill site:The mining of an unsanitary arid landfill

Landfilling is viewed nowadays as a serious threat associated with various burdens and stressors on the urban environment. To date, there is little information available on actual value of landfilled waste namely plastic solid waste (PSW) resulting from mining operations. In this work, PSW reclaimed from an active unsanitary landfill site (MAB) has been studied with the aim of determining its thermal profile and degradation behaviour for future utilisation in thermo-chemical conversion (TCC) processes. The materials were characterised by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) in accordance with internationally approved test methods in a simulated pyrolytic environment. In addition, chemical analysis using Fourier Transform Infrared Spectroscopy (FTIR) was applied to study the nature of the materials reclaimed. The degradation kinetics of the reclaimed PSW were studied with the aim of determining the apparent activation energy (Ea) of the pyrolytic reactions. The Ea values determined ranged from 199 to 266 kJ mol−1 which is in-line with pyrolytic reactions applicable for future use in fuel recovery units. TGA showed a clear shift in thermograms indicating a clear change in the degradation mechanism. The physico-chemical studies conducted on the materials also favours TCC treatment over other conventional end of life options such as physical (mechanical) recycling or incineration. The degradation mechanism was also determined from the Criado method showing that Avarami-Erofeve was the model that best represents PSW degradation. Overall, this work points towards future intervention schemes for reclaimed municipal solid waste (MSW) and in particular PSW favouring TCC technologies

Whole Genome Sequences of Three Treponema pallidum ssp. pertenue Strains: Yaws and Syphilis Treponemes Differ in Less than 0.2% of the Genome Sequence

Spirochete Treponema pallidum ssp. pertenue (TPE) is the causative agent of yaws while strains of Treponema pallidum ssp. pallidum (TPA) cause syphilis. Both yaws and syphilis are distinguished on the basis of epidemiological characteristics and clinical symptoms. Neither treponeme can reproduce outside the host organism, which precludes the use of standard molecular biology techniques used to study cultivable pathogens. In this study, we determined high quality whole genome sequences of TPE strains and compared them to known genetic information for T. pallidum ssp. pallidum strains. The genome structure was identical in all three TPE strains and also between TPA and TPE strains. The TPE genome length ranged between 1,139,330 bp and 1,139,744 bp. The overall sequence identity between TPA and TPE genomes was 99.8%, indicating that the two pathogens are extremely closely related. A set of 34 TPE genes (3.5%) encoded proteins containing six or more amino acid replacements or other major sequence changes. These genes more often belonged to the group of genes with predicted virulence and unknown functions suggesting their involvement in infection differences between yaws and syphilis

Role of zinc and α2macroglobulin on thymic endocrine activity and on peripheral immune efficiency (natural killer activity and interleukin 2) in cervical carcinoma

Decreased natural killer (NK) activity as well as interleukin 2 (IL-2) are risk factors for the progression of cervical carcinoma. NK activity and IL-2 may be thymus controlled. Plasma levels of active thymulin, a zinc-dependent thymic hormone (ZnFTS), are reduced in cancer because of the low peripheral zinc bioavailability. Zinc and thymulin are relevant for normal immune functions. α2-Macroglobulin is an inhibitor of matrix metalloproteases (MMPs) against invasive tumour proliferation. Because α2-macroglobulin has a binding affinity (Kd) for zinc that is higher than does thymulin, it may play a key role in immune efficiency in cancer. Plasma samples of 22 patients (age range 35–60 years) with locally advanced squamous cervical carcinoma and with FIGO stage Ib2–IIb were examined. They showed reduced active thymulin, decreased NK activity and IL-2 production, increased soluble IL-2 receptor (sIL-2R) and augmented α2-macroglobulin in the circulation, whereas plasma zinc levels were within the normal range for age. Significant positive correlations were found between zinc or active thymulin and α2-macroglobulin (r = 0.75, P< 0.01, r = 0.78, P< 0.01, respectively) in cancer patients. In vitro zinc increases IL-2 production from peripheral blood mononuclear cells (PBMCs) of cancer patients. These data suggest that an increase in α2-macroglobulin, which competes with thymulin for zinc binding, may be involved in causing a thymulin deficit with a consequent decrease of IL-2 and NK cytotoxicity. Thus, physiological zinc treatment in cervical carcinoma maybe restores impaired central and peripheral immune efficiency. © 1999 Cancer Research Campaig