STX13 regulates cargo delivery from recycling endosomes during melanosome biogenesis

Melanosomes are a class of lysosome-related organelles produced by melanocytes. Biogenesis of melanosomes requires the transport of melanin-synthesizing enzymes from tubular recycling endosomes to maturing melanosomes. The SNARE proteins involved in these transport or fusion steps have been poorly studied. We found that depletion of syntaxin 13 (STX13, also known as STX12), a recycling endosomal Qa-SNARE, inhibits pigment granule maturation in melanocytes by rerouting the melanosomal proteins such as TYR and TYRP1 to lysosomes. Furthermore, live-cell imaging and electron microscopy studies showed that STX13 co-distributed with melanosomal cargo in the tubular-vesicular endosomes that are closely associated with the maturing melanosomes. STX family proteins contain an N-terminal regulatory domain, and deletion of this domain in STX13 increases both the SNARE activity in vivo and melanosome cargo transport and pigmentation, suggesting that STX13 acts as a fusion SNARE in melanosomal trafficking pathways. In addition, STX13-dependent cargo transport requires the melanosomal R-SNARE VAMP7, and its silencing blocks the melanosome maturation, reflecting a defect in endosome-melanosome fusion. Moreover, we show mutual dependency between STX13 and VAMP7 in regulating their localization for efficient cargo delivery to melanosomes.

Haloarchaeal gas vesicle nanoparticles displaying Salmonella antigens as a novel approach to vaccine development

A safe, effective, and inexpensive vaccine against typhoid and other Salmonella diseases is urgently needed. In order to address this need, we are developing a novel vaccine platform employing buoyant, self-adjuvanting gas vesicle nanoparticles (GVNPs) from the halophilic archaeon Halobacterium sp. NRC-1, bioengineered to display highly conserved Salmonella enterica antigens. As the initial antigen for testing, we selected SopB, a secreted inosine phosphate effector protein injected by pathogenic S. enterica bacteria during infection into the host cells. Two highly conserved sopB gene segments near the 3'- region, named sopB4 and sopB5, were each fused to the grIpC gene, and resulting SopB-GVNPs were purified by centrifugally accelerated flotation. Display of SopB4 and SopB5 antigenic epitopes on GVNPs was established by Western blotting analysis using antisera raised against short synthetic peptides of SopB. Immunostimulatory activities of the SopB4 and B5 nanoparticles were tested by intraperitoneal administration of SopB-GVNPs to BALB/c mice which had been immunized with S. enterica serovar Typhimurium 14028 ApmrG-H111-D (DV-STM-07), a live attenuated vaccine strain. Proinflammatory cytokines IFN-y, IL-2, and IL-9 were significantly induced in mice boosted with SopB5-GVNPs, consistent with a robust Thl response. After challenge with virulent S. enterica serovar Typhimurium 14028, bacterial burden was found to be diminished in spleen of mice boosted with SopB4-GVNPs and absent or significantly diminished in liver, mesenteric lymph node, and spleen of mice boosted with SopB5GVNPs, indicating that the C-terminal portions of SopB displayed on GVNPs elicit a protective response to Salmonella infection in mice. SopB antigen-GVNPs were also found to be stable at elevated temperatures for extended periods without refrigeration. The results show that bioengineered GVNPs are likely to represent a valuable platform for antigen delivery and development of improved vaccines against Salmonella and other diseases.

Rab9A is required for delivery of cargo from recycling endosomes to melanosomes

Melanosomes are a type of lysosome-related organelle that is commonly defective in Hermansky-Pudlak syndrome. Biogenesis of melanosomes is regulated by BLOC-1, -2, -3, or AP-1, -3 complexes, which mediate cargo transport from recycling endosomes to melanosomes. Although several Rab GTPases have been shown to regulate these trafficking steps, the precise role of Rab9A remains unknown. Here, we found that a cohort of Rab9A associates with the melanosomes and its knockdown in melanocytes results in hypopigmented melanosomes due to mistargeting of melanosomal proteins to lysosomes. In addition, the Rab9A-depletion phenotype resembles Rab38/ 32-inactivated or BLOC-3-deficient melanocytes, suggesting that Rab9A works in line with BLOC-3 and Rab38/ 32 during melanosome cargo transport. Furthermore, silencing of Rab9A, Rab38/ 32 or its effector VARP, or BLOC-3-deficiency in melanocytes decreased the length of STX13-positive recycling endosomal tubules and targeted the SNARE to lysosomes. This result indicates a defect in directing recycling endosomal tubules to melanosomes. Thus, Rab9A and its co-regulatory GTPases control STX13-mediated cargo delivery to maturing melanosomes.

Chemical recycling of municipal packaging waste by pyrolysis

[EN]The present doctoral thesis centers on studying pyrolysis as a chemical recycling technique for rejected packaging waste fractions coming from separation and sorting plants. The pyrolysis experiments have been carried out in a lab-scale installation equipped with a 3.5 L semi-batch reactor and a condensation and collection system for the liquids and gases generated. In the present thesis, an experimental study on the conventional pyrolysis process applied to the aforementioned waste fractions has been conducted, as well as the study of non-conventional or advanced pyrolysis processes such as catalytic and stepwise pyrolysis. The study of the operating parameters has been carried out using a mixed plastics simulated sample, the composition of which is similar to that found in real fractions, and subsequently the optimized process has been applied to real packaging waste. An exhaustive characterization of the solids, liquids and gases obtained in the process has been made after each experiment and their potential uses have been established. Finally, an empirical model that will predict the pyrolysis yields (% organic liquid, % aqueous liquid, % gases, % char, % inorganic solid) as a function of the composition of the initial sample has been developed. As a result of the experimental work done, the requirements have been established for an industrial packaging waste pyrolysis plant that aims to be sufficiently versatile as to generate useful products regardless of the nature of the raw material.

Tracking the evolution of waste recycling research using overlay maps of science

Tracking the evolution of research in waste recycling science (WRS) can be valuable for environmental agencies, as well as for recycling businesses. Maps of science are visual, easily readable representations of the cognitive structure of a branch of science, a particular area of research or the global spectrum of scientific production. They are generally built upon evidence collected from reliable sources of information, such as patent and scientific publication databases. This study uses the methodology developed by Rafols et al. (2010) to make a “double overlay map” of WRS upon a basemap reflecting the cognitive structure of all journal-published science, for the years 2005 and 2010. The analysis has taken into account the cognitive areas where WRS articles are published and the areas from where it takes its intellectual nourishing, paying special attention to the growing trends of the key areas. Interpretation of results lead to the conclusion that extraction of energy from waste will probably be an important research topic in the future, along with developments in general chemistry and chemical engineering oriented to the recovery of valuable materials from waste. Agricultural and material sciences, together with the combined economics, politics and geography field, are areas with which WRS shows a relevant and ever increasing cognitive relationship.

Plan de Marketing de una PYME: Beotibar Recycling, S.L.

A la hora de entender y analizar lo que está pasando en su empresa, son muchas las dudas que les surgen a los directivos de estas pequeñas y medianas empresas cuando se dan cuenta de que no han desarrollado una estrategia adecuada a sus negocios o por otro lado, no han utilizado ninguna. Para solucionar este problema, existen varias técnicas y estrategias de marketing que se pueden desarrollar. Una de ellas es la elaboración del plan de marketing, que permitirá identificar los medios necesarios para conseguir alcanzar los objetivos ligados al potencial de la empresa. Pero, ¿Para qué sirve exactamente un plan de marketing? Gracias a él se determinarán las necesidades reales del mercado o los clientes potenciales, se desarrollarán y lanzarán productos y servicios de manera ordenada, se establecerá a través de que canales se introducirán en el mercado y se podrá identificar mejor la manera de comunicar nuestra marca/productos/servicios al mercado. En este trabajo se realizará un plan de marketing en la práctica para “Beotibar Recycling S.L”, una empresa perteneciente al sector del reciclado del papel. Siguiendo el método analítico-sintético, el contenido del proyecto constará de un análisis previo del mercado y un diagnóstico de situación. Posteriormente, se marcaran unos objetivos y se implementaran unas estrategias y acciones a seguir, para finalmente organizar un plan de seguimiento y control de estas últimas.

Efficient diode-pumped Tm : YALO(3) laser with a pump recycling scheme

By using a pump recycling configuration, the maximum power of 8.1 W in the wavelength range 1.935-1.938 mu m is generated by a 5-mm long Tm:YAlO3 (4 at. %) laser operating at 18 degrees C with a pump power of 24 W. The highest slope efficiency of 42% is attained, and the pump quantum efficiency is up to 100%. The Tm:YAlO3 laser is employed as a pumping source of singly-doped Ho(l%):GdVO4 laser operating at room temperature, in which continuous wave output power of greater than 0.2 W at 2.05 mu m is achieved with a slope efficiency of 9%.

Diode-pumped Q-switched Tm:YAP laser with a pump recycling scheme

We reported on a diode end-pumped AO Q-switched Tm:YAP laser at 1937 nm. The average output power was 3.9 W, with a slope efficiency of 29.4% and optical-optical conversion efficiency of 21.6% at a 5-kHz repetition rate. The temperature dependency of the output power and the pulse width at different repetition rates were investigated in details.