Lung cancer is the leading cause of cancer mortality worldwide and KRAS is the most commonly mutated gene in lung adenocarcinoma (LUAD). The 78-kDa glucose-regulated protein GRP78/BiP is a key ...endoplasmic reticulum chaperone protein and a major pro-survival effector of the unfolded protein response (UPR). Analysis of the Cancer Genome Atlas database and immunostain of patient tissues revealed that compared to normal lung, GRP78 expression is generally elevated in human lung cancers, including tumors bearing the KRAS
mutation. To test the requirement of GRP78 in human lung oncogenesis, we generated mouse models containing floxed Grp78 and Kras Lox-Stop-Lox G12D (Kras
) alleles. Simultaneous activation of the Kras
allele and knockout of the Grp78 alleles were achieved in the whole lung or selectively in lung alveolar epithelial type 2 cells known to be precursors for adenomas that progress to LUAD. Here we report that GRP78 haploinsufficiency is sufficient to suppress Kras
-mediated lung tumor progression and prolong survival. Furthermore, GRP78 knockdown in human lung cancer cell line A427 (Kras
) leads to activation of UPR and apoptotic markers and loss of cell viability. Our studies provide evidence that targeting GRP78 represents a novel therapeutic approach to suppress mutant KRAS-mediated lung tumorigenesis.
Micrurus dumerilii is a coral snake of clinic interest in Colombia. Its venom is mainly composed of phospholipases A2 being MdumPLA2 the most abundant protein. Nevertheless, Micrurus species produce ...a low quantity of venom, which makes it difficult to produce anticoral antivenoms. Therefore, in this work, we present the recombinant expression of MdumPLA2 to evaluate its biological activities and its immunogenic potential to produce antivenoms. For this, a genetic construct rMdumPLA2 was cloned into the pET28a vector and expressed heterologously in bacteria. His-rMdumPLA2 was extracted from inclusion bodies, refolded in vitro, and isolated using affinity and RP-HPLC chromatography. His-rMdumPLA2 was shown to have phospholipase A2 activity, a weak anticoagulant effect, and induced myonecrosis and edema. The anti-His-rMdumPLA2 antibodies produced in rabbits recognized native PLA2, the complete venom of M. dumerilii, and a phospholipase from another species of the Micrurus genus. Antibodies neutralized 100% of the in vitro phospholipase activity of the recombinant toxin and a moderate percentage of the myotoxic activity of M. dumerilii venom in mice. These results indicate that His-rMdumPLA2 could be used as an immunogen to improve anticoral antivenoms development. This work is the first report of an M. dumerilii functional recombinant PLA2.
The purpose of this paper consists of implementing an educational board game to evaluate the information security awareness level of healthcare personnel. The National Health Service Greater Glasgow ...and Clyde (NHSGGC) Information Security Acceptable Use Policy was used as a basis to generate the educational content of the board game and Lev Vygotsky’s social development theory was followed for the learning process of the participants. Two evaluations were carried out during this study. The results obtained during the first evaluation showed that it is fundamental to design the board game based on a set of rules in information security enacted by an organization to properly guide the participants with the knowledge they need to counter security incidents. The second evaluation showed that redesigning the content of the board game based on the information security policies of the NHSGGC, resulted in a more effective way of guiding participants on the procedures required for compliance with the policies of this health institution and offer them an understanding of the risks behind security incidents. This was demonstrated during this evaluation since the results obtained gave an approximation that it is possible to increase the level of awareness of information security in people regardless of their area of work or studies.
El policultivo de camarón-tilapia es una de las estrategias de mitigación para los impactos sanitarios y los efluentes en las granjas camaroneras. Su finalidad principal es incrementar la producción ...y controlar la calidad del agua de sus efluentes. En casos de éxito se controlaron algunas enfermedades y mejoró la calidad del suelo en estanques de tierra. En esta revisión, se analizan los beneficios que se obtienen en la calidad del agua, sedimento, medio ambiente, incremento del rendimiento y el mejoramiento de los parámetros zootécnicos de camarones co-cultivados con tilapia. Los policultivos de camarón-tilapia pueden contribuir con un efecto positivo sobre la calidad del agua y sedimentos de los estanques y efluentes de las granjas de cultivo, lo cual depende de las condiciones de densidad y ambiente donde se desarrollan. Sin embargo, se requiere adaptar y demostrar su funcionalidad y rentabilidad comercial a pequeña escala e industrialmente.
Abstract Autoantibodies against SCLC-associated neuronal antigen ELAVL4 (HuD) have been linked to smaller tumors and improved survival, but the antigenic epitope and mechanism of autoimmunity have ...never been solved. We report that recombinant human ELAVL4 protein incubated under physiological conditions acquires isoaspartylation, a type of immunogenic protein damage. Specifically, the N-terminal region of ELAVL4, previously implicated in SCLC-associated autoimmunity, undergoes isoaspartylation in vitro , is recognized by sera from anti-ELAVL4 positive SCLC patients and is highly immunogenic in subcutaneously injected mice and in vitro stimulated human lymphocytes. Our data suggest that isoaspartylated ELAVL4 is the trigger for the SCLC-associated anti-ELAVL4 autoimmune response.
This article contains experimental data examining the propensity of neuronal ELAVL proteins to become isoaspartylated. The data are related to the article “Isoaspartylation appears to trigger small ...cell lung cancer-associated autoimmunity against neuronal protein ELAVL4” (M.A. Pulido, M.K. DerHartunian, Z. Qin, E.M. Chung, D.S. Kang, A.W. Woodham, J.A. Tsou, R. Klooster, O. Akbari, L. Wang, W.M. Kast, S.V. Liu, J.J.G.M. Verschuuren, D.W. Aswad, I.A. Laird-Offringa, 2016) 1, in which it was reported that the N-terminal region of recombinant human ELAVL4 protein, incubated under physiological conditions, acquires a type of highly immunogenic protein damage. Here, we present Western blot analysis data generated by using an affinity-purified polyclonal rabbit antibody (raised against an N-terminal ELAVL4 isoaspartyl-converted peptide) to probe recombinant protein fragments of the other three members of the ELAVL family: the highly homologous neuronal ELAVL2 (HuB) and ELAVL3 (HuC), and the much less homologous ubiquitously expressed ELAVL1 (HuR).
Micrurus dumerilii is a coral snake of clinic interest in Colombia. Its venom is mainly composed of phospholipases Asub.2 being MdumPLAsub.2 the most abundant protein. Nevertheless, Micrurus species ...produce a low quantity of venom, which makes it difficult to produce anticoral antivenoms. Therefore, in this work, we present the recombinant expression of MdumPLAsub.2 to evaluate its biological activities and its immunogenic potential to produce antivenoms. For this, a genetic construct rMdumPLAsub.2 was cloned into the pET28a vector and expressed heterologously in bacteria. His-rMdumPLAsub.2 was extracted from inclusion bodies, refolded in vitro, and isolated using affinity and RP-HPLC chromatography. His-rMdumPLAsub.2 was shown to have phospholipase Asub.2 activity, a weak anticoagulant effect, and induced myonecrosis and edema. The anti-His-rMdumPLAsub.2 antibodies produced in rabbits recognized native PLAsub.2 , the complete venom of M. dumerilii, and a phospholipase from another species of the Micrurus genus. Antibodies neutralized 100% of the in vitro phospholipase activity of the recombinant toxin and a moderate percentage of the myotoxic activity of M. dumerilii venom in mice. These results indicate that His-rMdumPLAsub.2 could be used as an immunogen to improve anticoral antivenoms development. This work is the first report of an M. dumerilii functional recombinant PLAsub.2 .
is a coral snake of clinic interest in Colombia. Its venom is mainly composed of phospholipases A
being MdumPLA
the most abundant protein. Nevertheless,
species produce a low quantity of venom, which ...makes it difficult to produce anticoral antivenoms. Therefore, in this work, we present the recombinant expression of MdumPLA
to evaluate its biological activities and its immunogenic potential to produce antivenoms. For this, a genetic construct rMdumPLA
was cloned into the pET28a vector and expressed heterologously in bacteria. His-rMdumPLA
was extracted from inclusion bodies, refolded in vitro, and isolated using affinity and RP-HPLC chromatography. His-rMdumPLA
was shown to have phospholipase A
activity, a weak anticoagulant effect, and induced myonecrosis and edema. The anti-His-rMdumPLA
antibodies produced in rabbits recognized native PLA
, the complete venom of
, and a phospholipase from another species of the
genus. Antibodies neutralized 100% of the in vitro phospholipase activity of the recombinant toxin and a moderate percentage of the myotoxic activity of
venom in mice. These results indicate that His-rMdumPLA
could be used as an immunogen to improve anticoral antivenoms development. This work is the first report of an
functional recombinant PLA
.
Biomass is a promising source to produce energy or biofuels, chemicals and biobased products in order to reduce dependency on fossil fuels. Fast growing crops are an ideal source of lignocellulosic ...biomass due to their high cellulose, hemicellulose and lignin content. In this sense, eleven (11) families were selected and compiled in twenty one (21) species according to soil type and crop requirement. Cultivation areas were selected through analyses of the Geographic Information System (SIG by its Spanish acronym) highlighting soil characteristics (mainly as arid and semiarid areas) and crop requirements. Two main groups were defined during this analysis: species established in Colombia and species not yet reported in Colombia. Species established in Colombia were related to family Asteraceae, Euphorbiaceae, Fabaceae, Lamiáceas, Myrtaceae and Pinaceae. The estimated area for this crop group was around 1.7 million hectares with an energy potential of 94,677 TJ/year. Meanwhile, species not yet reported in Colombia were related to family Leguminosae, Myrtaceae, Paulowniaceae, Pinaceae and Poaceae. The estimated area for this crop group was around 2.4 million of hectares with an energy potential of 65,525 TJ/year. The energy value achieved for each specie was in the range of 2,000 TJ/year to 10,000 TJ/year, the other species that were out of this range, were not take into account. The main region available to develop these types of crops in Colombia was located in the departments of Cundinamarca, Antioquia, Santander, Quindío, Risaralda, Caldas, Tolima, Valle del Cauca and Nariño. Technologies such as gasification and pyrolysis were identified to be applied to some families as Asteraceae, Euphorbiaceae, Paulowniaceae and Pinaceae, which have around 25% of lignin; obtaining liquid and gaseous fuels. Meanwhile, a high cellulose and hemicellulose content (48% and 64% respectively) in families as Fabaceae, Lamiáceas and Myrtaceae make them viable to be used as a sugar source in biofuel production.
Abstract
Patients with small-cell lung cancer (SCLC), a highly metastatic type of lung cancer, show a dismal 5-year survival of only 5%. Effective therapies are urgently needed. Notably, about ...fifteen-percent of SCLC-patients harbor anti-Hu autoantibodies, which are associated with improved survival. This suggests that the anti-Hu response might be harnessed for immunotherapy. A conditional SCLC mouse model with floxed Rb1 and p53 alleles, in which the cancer is induced by instilling Cre-recombinase into the lungs via intratracheal intubation, offers a unique opportunity to study the immune response in detail. We have previously shown that, just like human SCLC patients, a fraction of these SCLC-carrying mice show an anti-Hu autoantibody response. Here we examine the presence of anti-Hu reactive T-cells using a prospective cohort (n=45) of SCLC-induced mice. All mice develop SCLC, showing systemic disease in 4-7 months following inactivation of floxed Rb1 and p53 alleles. Monthly autoantibody analysis by western blotting revealed an antibody response in a subset of mice, sometimes as early as 1 month after induction. Reactivity usually peaked approximately 3-4 months after induction and declined as disease progressed. This suggests an immune escape mechanism and/or antibody adsorption by the rapidly proliferating tumors. T-cells were isolated at euthanasia from the spleens of moribund mice. Proliferating anti-HuD CD4-positive T-cells were detected in fifty-one-percent of mice (eighteen out of thirty-five) by flow cytometry. Anti-HuD T-cell proliferation from spleen cells was validated in a subsequent prospective cohort (n=45) of SCLC-induced mice using thymidine incorporation. Stratifying for antibody positive and negative mice using an stringent cutoff, there was no significant difference in T cell response between the two groups (p=0.11), however four mice in the antibody positive group showed the most elevated anti-HuD T-cell response. Our observations warrant a much larger study, in which in-depth characterization of the humoral and cellular the anti-HuD immune response is carried out at multiple time points and correlated with a detailed analysis of the stage of tumor development and mestastasis. Preliminary data hints at a negative correlation between the anti-HuD antibody and T-cell response on one hand and tumor size and metastasis on the other. The observation that the Hu antigen in mice with SCLC is a target for both humoral and cellular immune responses underscores the utility of this mouse model in the preclinical evaluation the anti-Hu immune response for therapy and clinical translation.
Citation Format: Mario A. Pulido, Vincent Lombardi, Diane Lee, Yiwei Wang, Eric Chung, Lina Wang, W. Martin Kast, Omid Akbari, Ite A. Laird-Offringa. Fluctuating antibody response and CD4-positive T-cells in a small-cell lung cancer mouse model. abstract. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3623. doi:10.1158/1538-7445.AM2014-3623
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