Proteomic and functional analysis identifies galectin-1 as a novel regulatory component of the cytotoxic granule machinery

Secretory granules released by cytotoxic T lymphocytes (CTLs) are powerful weapons against intracellular microbes and tumor cells. Despite significant progress, there is still limited information on the molecular mechanisms implicated in target-driven degranulation, effector cell survival and composition and structure of the lytic granules. Here, using a proteomic approach we identified a panel of putative cytotoxic granule proteins, including some already known granule constituents and novel proteins that contribute to regulate the CTL lytic machinery. Particularly, we identified galectin-1 (Gal1), an endogenous immune regulatory lectin, as an integral component of the secretory granule machinery and unveil the unexpected function of this lectin in regulating CTL killing activity. Mechanistic studies revealed the ability of Gal1 to control the non-secretory lytic pathway by influencing Fas–Fas ligand interactions. This study offers new insights on the composition of the cytotoxic granule machinery, highlighting the dynamic cross talk between secretory and non-secretory pathways in controlling CTL lytic function

Dysregulation of specialized delay/interference-dependent working memory following loss of dysbindin-1A in schizophrenia-related phenotypes

Dysbindin-1, a protein that regulates aspects of early and late brain development, has been implicated in the pathobiology of schizophrenia. As the functional roles of the three major isoforms of dysbindin-1, (A, B, and C) remain unknown, we generated a novel mutant mouse, dys-1A -/-, with selective loss of dysbindin-1A and investigated schizophrenia-related phenotypes in both males and females. Loss of dysbindin-1A resulted in heightened initial exploration and disruption in subsequent habituation to a novel environment, together with heightened anxiety-related behavior in a stressful environment. Loss of dysbindin-1A was not associated with disruption of either long-term (olfactory) memory or spontaneous alternation behavior. However, dys-1A -/-showed enhancement in delay-dependent working memory under high levels of interference relative to controls, ie, impairment in sensitivity to the disruptive effect of such interference. These findings in dys-1A -/-provide the first evidence for differential functional roles for dysbindin-1A vs dysbindin-1C isoforms among phenotypes relevant to the pathobiology of schizophrenia. Future studies should investigate putative sex differences in these phenotypic effects

The identification and functional characterisation of caspase substrates involved in inflammation

THESIS 9354During inflammation the immune system responds to harmful stimuli, tries to control the stimuli and initiate a healing process for damaged tissue. After recognition of microbes by the innate immune system caspases are activated and are required to activate cytokines. Human caspases have been studied for over two decades with most work in the field focused on apoptotic caspases. As a result hundreds of apoptotic caspase substrates have been identified to date, but only 2 inflammatory caspase substrates have been confirmed. Identification of novel targets for the caspases activated during the inflammatory process could provide new insights into the molecular mechanism of host defense, identify important immuno-regulatory molecules and targets for anti-inflammatory drug development

Suppression of Interleukin-33 Bioactivity through Proteolysis by Apoptotic Caspases

Interleukin-33 (IL-33) is a member of the IL-1 family and is involved in polarization of T cells toward a T helper 2 (Th2) cell phenotype. IL-33 is thought to be activated via caspase-1-dependent proteolysis, similar to the proinflammatory cytokines IL-1? and IL-18, but this remains unproven. Here we showed that IL-33 was processed by caspases activated during apoptosis (caspase-3 and -7) but was not a physiological substrate for caspases associated with inflammation (caspase-1, -4, and -5). Furthermore, caspase-dependent processing of IL-33 was not required for ST2 receptor binding or ST2-dependent activation of the NF-?B transcription factor. Indeed, caspase-dependent proteolysis of IL-33 dramatically attenuated IL-33 bioactivity in vitro and in vivo. These data suggest that IL-33 does not require proteolysis for activation, but rather, that IL-33 bioactivity is diminished through caspase-dependent proteolysis within apoptotic cells. Thus, caspase-mediated proteolysis acts as a switch to dampen the proinflammatory properties of IL-33

Assessing Micro-Hydropower in Himachal Pradesh

Himachal Pradesh is the largest producer of hydroelectricity in India, and boasts a 99.8% rural electrification rate. However, micro-hydropower is electrical power with off-grid applications that does not cause the negative impacts of large-scale projects. Our team researched the viability of using micro-hydro turbines to power individual households. We conducted interviews with 39 people across eight locations to understand the electrical needs of the community as wells as opinions on micro-hydro projects. We then constructed a micro-turbine prototype and tested it at a local river site. Finally, we made several recommendations for improvements to our prototype as well as continued research on the feasibility of micro-hydro technologies

Creating a reference database of cargo inspection X-ray images using high energy CT of cargo mock-ups

International audienceCustoms continue to use a wide range of technology in protecting against terrorism and the movement of illicit trade and prohibited imports. The throughput of scanned vehicles and cargo increases and just keeps on growing. Therefore, the need of automated algorithms to help screening officers in inspection, examination or surveillance of vehicles and containers is crucial. In this context, the successful collaboration between manufacturers and customs offices is of key importance. Facing this topic, within the seventh framework program of the European Commission, the project ACXIS “Automated Comparison of X-ray Images for cargo Scanning” arose. This project develops a reference database for X-ray images of illegal and legitimate cargo, procedures and algorithms to uniform X-ray images of different cargo scanners, and an automated identification of potentially illegal cargo

Proteomic and functional analysis identifies galectin-1 as a novel regulatory component of the cytotoxic granule machinery

Secretory granules released by cytotoxic T lymphocytes (CTLs) are powerful weapons against intracellular microbes and tumor cells. Despite significant progress, there is still limited information on the molecular mechanisms implicated in target-driven degranulation, effector cell survival and composition and structure of the lytic granules. Here, using a proteomic approach we identified a panel of putative cytotoxic granule proteins, including some already known granule constituents and novel proteins that contribute to regulate the CTL lytic machinery. Particularly, we identified galectin-1 (Gal1), an endogenous immune regulatory lectin, as an integral component of the secretory granule machinery and unveil the unexpected function of this lectin in regulating CTL killing activity. Mechanistic studies revealed the ability of Gal1 to control the non-secretory lytic pathway by influencing Fas?Fas ligand interactions. This study offers new insights on the composition of the cytotoxic granule machinery, highlighting the dynamic cross talk between secretory and non-secretory pathways in controlling CTL lytic function.Fil: Clemente, Tiago. Universidade de Sao Paulo; Brasil. Instituto Nacional de Ciência e Tecnologia ; BrasilFil: Vieira, Narcisio J.. Universidade de Sao Paulo; Brasil. Instituto Nacional de Ciência e Tecnologia ; BrasilFil: Cerliani, Juan Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Adrain, Colin. Instituto Gulbenkian de Ciência; PortugalFil: Luthi, Alexander. Trinity College; IrlandaFil: Dominguez, Mariana. Universidade de Sao Paulo; BrasilFil: Yon, Monica. Universidade de Sao Paulo; BrasilFil: Barrence, Fernanda C.. Universidade de Sao Paulo; BrasilFil: Riul, Thalita B.. Universidade de Sao Paulo; BrasilFil: Cummings, Richard D.. Harvard Medical School; Estados UnidosFil: Zorn, Thelma. Universidade de Sao Paulo; BrasilFil: Amigorena, Sebastian. Inserm; FranciaFil: Dias Baruffi, Marcelo. Universidade de Sao Paulo; BrasilFil: Rodriguez, Mauricio M.. Universidade de Sao Paulo; BrasilFil: Seamus, J. Martin. Trinity College; IrlandaFil: Rabinovich, Gabriel Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Amarante Mendez, Gustavo P.. Trinity College; Irlanda. Universidade de Sao Paulo; Brasil. Instituto Nacional de Ciência e Tecnologia ; Brasi