The Role of Cancer-Associated Fibroblasts in Lung Tumorigenesis

The extracellular milieu is rich in growth factors that drive tumor progression,but the mechanisms that govern tumor cell sensitivity to those ligands have notbeen fully defined. In this study, we address this question in mice that developmetastatic lung adenocarcinomas through the suppression of the microRNA-200 (miR-200) family. Cancer-associated fibroblasts (CAF) enhance tumorgrowth and invasion by secreting VEGF-A that binds to VEGFR1, a processrequired for tumor growth and metastasis in mice and correlated with a poorprognosis in lung adenocarcinoma patients. In this study, we discovered thatmiR-200 blocked CAF-induced tumor cell invasion by directly targetingVEGFR1 in tumor cells. In the context of previous studies, our findings suggestthat the miR-200 family is a point of convergence for diverse biologic processesthat regulate tumor cell proliferation, invasion, and metastasis; its target genesixdrive epithelial-to-mesenchymal transition (ZEB1 and ZEB2) and promotesensitivity to a potent tumor growth factor emanating from the microenvironment(VEGFR1). Clinical trials should focus not only on the role of VEGFR1 inangiogenesis but also on the expression and activation of VEGFR1 in tumorcells by stromal sources of VEGF-A in the tumor microenvironment as a targetfor metastasis prevention.

Suppression of T lymphocyte activation in simulated microgravity

Immune dysfunction is encountered during spaceflight. Various aspects of spaceflight, including microgravity, cosmic radiation, and both physiological and psychological stress, may perturb immune function. We sought to understand the impact of microgravity alone on the cellular mechanisms critical to immunity. Clinostatic RWV bioreactors that simulate aspects of microgravity were used to analyze the response of human PBMC to polyclonal and oligoclonal activation. PHA responsiveness in the RWV bioreactor was almost completely diminished. IL-2 and IFN-$\gamma$ secretion was reduced whereas IL-1$\beta$ and IL-6 secretion was increased, suggesting that monocytes may not be as adversely affected by simulated microgravity as T cells. Activation marker expression (CD25, CD69, CD71) was significantly reduced in RWV cultures. Furthermore, addition of exogenous IL-2 to these cultures did not restore proliferation. Antigen specific T cell activation, including the mixed-lymphocyte reaction, tetanus toxoid responsiveness, and Borrelia activation of a specific T cell line, was also suppressed in the RWV bioreactor.^ The role of altered culture conditions in the suppression of T cell activation were considered. Potential reduced cell-cell and cell-substratum interactions in the RWV bioreactor may play a role in the loss of PHA responsiveness. However, PHA activation in Teflon culture bags that limit cell-substratum interactions was not affected. Furthermore, increasing cell-population density, and therefore cell-cell interactions, in the RWV cultures did not help restore PHA activation. However, placing PBMC within small collagen beads did partially restore PHA responsiveness. Finally, activation of purified T cells with crosslinked CD2/CD28 or CD3/CD28 antibody pairs, which does not require costimulation through cell-cell contact, was completely suppressed in the RWV bioreactor suggesting a defect internal to the T cell.^ Activation of both PBMC and purified T cells with PMA and ionomycin was unaffected by RWV culture, indicating that signaling mechanisms downstream of PKC activation and calcium flux are not sensitive to simulated microgravity. Furthermore, sub-mitogenic doses of PMA alone but not ionomycin alone restored PHA responsiveness of PBMC in RWV culture. Thus, our data indicate that during polyclonal activation in simulated microgravity, there is a specific dysfunction within the T cell involving the signaling pathways upstream of PKC activation. ^

Review of Clément Vidal, "The Beginning and the End. The Meaning of Life in a Cosmological Perspective"

No more ground control. Dissertations ideally always have been ambitious, and this one can’t claim to be too humble in these regards. Together the pieces of this work form a world view that the author confesses to finding ‘magnificent’. The story he tells is one in which intelligence and complexity are keys to unlock the universe’s mysteries. Vidal tackles questions like ‘What is philosophy?’, ‘Where does it all come from?’, Where are we going?’, ‘Are we alone in the universe?’ and even ‘What is good and what is evil?’. Questions which philosophers for ages have tried to answer.

The role of nitric oxide in small intestinal motility in a model of acute inflammation

Motility responses of the small intestine of iNOS deficient mice (iNOS −/−) and their wildtype littermates (iNOS+/+) to the inflammatory challenge of lipopolysaccharide (LPS) were investigated. LPS administration failed to attenuate intestinal transit in iNOS−/− mice but depressed transit in their iNOS+/+ littermates. Supporting an inhibitory role for sustained nitric oxide (NO) synthesis in the regulation of intestinal motility during inflammation, iNOS immunoreactivity was upregulated in all regions of the small intestine of iNOS+/+ mice. In contrast, neuronal NOS was barely affected. Cyclooxygenase activation was determined by prostaglandin E2 (PGE2) concentration. Following LPS challenge, PGE2 levels were elevated in all intestinal segments in both animal groups. Moreover, COX-1 and COX-2 protein levels were elevated in iNOS+/+ mice in response to LPS, while COX-2 levels were similarly increased in iNOS −/− intestine. However, no apparent relationship was observed between increased prostaglandin concentrations and attenuated intestinal transit. The presence of heme oxygenase 1 (HO-1) in the murine small intestine was also investigated. In both animal groups HO-1 immunoreactivity in the proximal intestine increased in response to treatment, while the constitutive protein levels detected in the middle and distal intestine were unresponsive to LPS administration. No apparent correlation of HO-1 to the suppression of small intestinal motility induced by LPS administration was detected. The presence of S-nitrosylated contractile proteins in the small intestine was determined. γ-smooth muscle actin was basally nitrosylated as well as in response to LPS, but myosin light chain kinase and myosin regulatory chain (MLC20) were not. In conclusion, in a model of acute intestinal inflammation, iNOS-produced NO plays a significant role in suppressing small intestinal motility while nNOS, COX-1, COX-2 and HO-1 do not participate in this event. S-nitrosylation of γ-smooth muscle actin is associated with elevated levels of nitric oxide in the smooth muscle of murine small intestine. ^

The role of suppressor factors in the regulation of immune responses by ultraviolet radiation -induced suppressor T lymphocytes

The purpose of these studies was to determine the role of suppressor factors (TsF) in the regulation of immune responses by ultraviolet radiation-induced suppressor T lymphocytes (Ts). The Ts were induced following epicutaneous sensitization with contact allergens to an unirradiated site on mice irradiated five days earlier with 40 kJ/m$\sp2$ UVB (280-320 nm) radiation. The spleens of such mice contain afferent, hapten-specific, Thy-1$\sp+$, Lyt-1$\sp+$,2$\sp-$ Ts that suppress in vivo contact hypersensitivity (CHS) and antibody responses and the in vitro generation of cytotoxic T lymphocytes (CTL). Four approaches were used to determine the role of TsF. First, lysates produced from sonically-disrupted Ts were injected i.v. into normal animals; they inhibited CHS in vivo in a nonspecific manner. The lysates suppressed the induction and elicitation of CHS, and they inhibited the in vitro generation of CTL. Lysates prepared from splenocytes obtained from unirradiated mice or UV-irradiated, unsensitized mice failed to inhibit either response. Second, supernatants from cultures containing Ts, normal syngeneic responder lymphocytes, and hapten-modified stimulator cells were injected i.v. into normal recipients. They inhibited the induction of CHS and did so in a hapten-specific manner. Cellular and kinetic requirements were observed for the generation of suppressive activity. Splenocytes from mice treated with Ts supernatants suppressed CHS when transferred into normal animals. The supernatants also suppressed the in vitro generation of specific CTL. Third, the TsF-specific B16G monoclonal antibody was tested for its ability to modulate the effects of UV radiation in vivo. The i.v. injection of B16G into UV-irradiated mice reduced the suppression of CHS. Splenocytes of B16G-treated mice transferred into normal recipients, and they suppressed CHS, indicating that the Ts were not depleted. Fourth, B16G was used to isolate a putative TsF by antibody immunoadsorbance. When the B16G-bound fraction was eluted and injected i.v. into normal animals, it suppressed CHS and represented a 900-fold enrichment of activity over the starting material, based on specific activity. By SDS-PAGE, the B16G-bound material contained nondisulfide-linked 45- and 50-kDa components. These results suggest that TsF may play an immunoregulatory role in CHS. The isolation of a UV radiation-induced TsF lends credence to the involvement of such molecules. ^

The observatory of intelligence and territorial development, tool for innovation in processes of territorial planning in municipalities of the province of Jujuy

El municipio es considerado como un espacio donde sus habitantes comparten no sólo el territorio sino también los problemas y los recursos existentes. La institución municipal -como gobierno local- es el ámbito en el cual se toman decisiones sobre el territorio, que implican a sus habitantes. En cuanto a los actores, estos pueden ser funcionarios, empleados y la comunidad (individual y organizada en ongs), todos aportan sus conocimientos y valores, pero tienen diferentes intereses y diferentes tiempos. Vinculada a las decisiones, encontramos que la forma en que se gestiona la información territorial, es determinante si se pretende apuntar hacia acciones con impacto positivo, y sustentables en lo ambiental y en el tiempo. Este trabajo toma tres municipios: San Salvador de Jujuy, capital de la provincia localizada en los Valles Templados; San Pedro de Jujuy, principal municipio de la región de las Yungas y Tilcara en la Quebrada de Humahuaca. El aporte de la Inteligencia Territorial, a través del observatorio OIDTe, permite analizar los modos de gestión de la información, especialmente mediante el uso de las tecnologías de la información y comunicación (pagina web municipal, equipamiento informático en las oficinas, estrategias de comunicación y vinculación con la población) y mediante la organización de las estructuras administrativas (organigrama) por las cuales circula la información municipal. Además, con la participación enriquecedora de equipos multidisciplinarios en las diferentes etapas. Se busca, a partir de un diagnóstico, generar estrategias para la introducción de innovaciones con los propios actores municipales, a partir de las situaciones y modos culturales propios de cada lugar, incorporando los marcos conceptuales de la Inteligencia Territorial. En este sentido el OIDTe al promover el entendimiento entre los actores, institucionales y la sociedad, facilita la coordinación de diferentes intereses propiciando la toma de decisiones por acuerdos. Asimismo, el método Portulano, puede orientar la introducción de innovaciones en la coordinación de la información cartográfica, para que las diferentes oficinas puedan complementar sus aportes y la comunicación hacia fuera de la institución. En la fase de diagnóstico, se aplicaron entrevistas a informantes claves, se realizó un workshop con técnicos de planta permanente y funcionarios de áreas que manejan información territorial, y de planificación. También por la importancia de la capacidad instalada de recursos humanos, se analizó el nivel de instrucción y la capacitación con que cuenta el personal de planta permanente de cada área

The observatory of intelligence and territorial development, tool for innovation in processes of territorial planning in municipalities of the province of Jujuy

El municipio es considerado como un espacio donde sus habitantes comparten no sólo el territorio sino también los problemas y los recursos existentes. La institución municipal -como gobierno local- es el ámbito en el cual se toman decisiones sobre el territorio, que implican a sus habitantes. En cuanto a los actores, estos pueden ser funcionarios, empleados y la comunidad (individual y organizada en ongs), todos aportan sus conocimientos y valores, pero tienen diferentes intereses y diferentes tiempos. Vinculada a las decisiones, encontramos que la forma en que se gestiona la información territorial, es determinante si se pretende apuntar hacia acciones con impacto positivo, y sustentables en lo ambiental y en el tiempo. Este trabajo toma tres municipios: San Salvador de Jujuy, capital de la provincia localizada en los Valles Templados; San Pedro de Jujuy, principal municipio de la región de las Yungas y Tilcara en la Quebrada de Humahuaca. El aporte de la Inteligencia Territorial, a través del observatorio OIDTe, permite analizar los modos de gestión de la información, especialmente mediante el uso de las tecnologías de la información y comunicación (pagina web municipal, equipamiento informático en las oficinas, estrategias de comunicación y vinculación con la población) y mediante la organización de las estructuras administrativas (organigrama) por las cuales circula la información municipal. Además, con la participación enriquecedora de equipos multidisciplinarios en las diferentes etapas. Se busca, a partir de un diagnóstico, generar estrategias para la introducción de innovaciones con los propios actores municipales, a partir de las situaciones y modos culturales propios de cada lugar, incorporando los marcos conceptuales de la Inteligencia Territorial. En este sentido el OIDTe al promover el entendimiento entre los actores, institucionales y la sociedad, facilita la coordinación de diferentes intereses propiciando la toma de decisiones por acuerdos. Asimismo, el método Portulano, puede orientar la introducción de innovaciones en la coordinación de la información cartográfica, para que las diferentes oficinas puedan complementar sus aportes y la comunicación hacia fuera de la institución. En la fase de diagnóstico, se aplicaron entrevistas a informantes claves, se realizó un workshop con técnicos de planta permanente y funcionarios de áreas que manejan información territorial, y de planificación. También por la importancia de la capacidad instalada de recursos humanos, se analizó el nivel de instrucción y la capacitación con que cuenta el personal de planta permanente de cada área

The observatory of intelligence and territorial development, tool for innovation in processes of territorial planning in municipalities of the province of Jujuy

El municipio es considerado como un espacio donde sus habitantes comparten no sólo el territorio sino también los problemas y los recursos existentes. La institución municipal -como gobierno local- es el ámbito en el cual se toman decisiones sobre el territorio, que implican a sus habitantes. En cuanto a los actores, estos pueden ser funcionarios, empleados y la comunidad (individual y organizada en ongs), todos aportan sus conocimientos y valores, pero tienen diferentes intereses y diferentes tiempos. Vinculada a las decisiones, encontramos que la forma en que se gestiona la información territorial, es determinante si se pretende apuntar hacia acciones con impacto positivo, y sustentables en lo ambiental y en el tiempo. Este trabajo toma tres municipios: San Salvador de Jujuy, capital de la provincia localizada en los Valles Templados; San Pedro de Jujuy, principal municipio de la región de las Yungas y Tilcara en la Quebrada de Humahuaca. El aporte de la Inteligencia Territorial, a través del observatorio OIDTe, permite analizar los modos de gestión de la información, especialmente mediante el uso de las tecnologías de la información y comunicación (pagina web municipal, equipamiento informático en las oficinas, estrategias de comunicación y vinculación con la población) y mediante la organización de las estructuras administrativas (organigrama) por las cuales circula la información municipal. Además, con la participación enriquecedora de equipos multidisciplinarios en las diferentes etapas. Se busca, a partir de un diagnóstico, generar estrategias para la introducción de innovaciones con los propios actores municipales, a partir de las situaciones y modos culturales propios de cada lugar, incorporando los marcos conceptuales de la Inteligencia Territorial. En este sentido el OIDTe al promover el entendimiento entre los actores, institucionales y la sociedad, facilita la coordinación de diferentes intereses propiciando la toma de decisiones por acuerdos. Asimismo, el método Portulano, puede orientar la introducción de innovaciones en la coordinación de la información cartográfica, para que las diferentes oficinas puedan complementar sus aportes y la comunicación hacia fuera de la institución. En la fase de diagnóstico, se aplicaron entrevistas a informantes claves, se realizó un workshop con técnicos de planta permanente y funcionarios de áreas que manejan información territorial, y de planificación. También por la importancia de la capacidad instalada de recursos humanos, se analizó el nivel de instrucción y la capacitación con que cuenta el personal de planta permanente de cada área

Very Chicago : Mary Borden and the Art of Fiction

Peer reviewed

Three distinct domains of SSI-1/SOCS-1/JAB protein are required for its suppression of interleukin 6 signaling

Cytokine-inducible protein SSI-1 [signal transducers and activators of transcription (STAT)-induced STAT inhibitor 1, also referred to as SOCS-1 (suppressor of cytokine signaling 1) or JAB (Janus kinase-binding protein)] negatively regulates cytokine receptor signaling by inhibition of JAK kinases. The SSI family of proteins includes eight members that are structurally characterized by an SH2 domain and a C-terminal conserved region that we have called the SC-motif. In this study, we investigated the roles of these domains in the function of SSI-1. Results of reporter assays demonstrated that the pre-SH2 domain (24 aa in front of the SH2 domain) and the SH2 domain of SSI-1 were required for the suppression by SSI-1 of interleukin 6 signaling. Coexpression studies of COS7 cells revealed that these domains also were required for inhibition of three JAKs (JAK1, JAK2, and TYK2). Furthermore, deletion of the SH2 domain, but not the pre-SH2 domain, resulted in loss of association of SSI-1 with TYK2. Thus, SSI-1 associates with JAK family kinase via its SH2 domain, and the pre-SH2 domain is required for the function of SSI-1. Deletion of the SC-motif markedly reduced expression of SSI-1 protein in M1 cells, and this reduction was reversed by treatment with proteasome inhibitors, suggesting that this motif is required to protect the SSI-1 molecule from proteolytic degradation. Based on these findings, we concluded that three distinct domains of SSI-1 (the pre-SH2 domain, the SH2 domain, and the SC-motif) cooperate in the suppression of interleukin 6 signaling.

Monitoring the structure of Escherichia coli RNase P RNA in the presence of various divalent metal ions

Lead(II)-induced cleavage can be used as a tool to probe conformational changes in RNA. In this report, we have investigated the conformation of M1 RNA, the catalytic subunit of Escherichia coli RNase P, by studying the lead(II)-induced cleavage pattern in the presence of various divalent metal ions. Our data suggest that the overall conformation of M1 RNA is very similar in the presence of Mg2+, Mn2+, Ca2+, Sr2+ and Ba2+, while it is changed compared to the Mg2+-induced conformation in the presence of other divalent metal ions, Cd2+ for example. We also observed that correct folding of some M1 RNA domains is promoted by Pb2+, while folding of other domain(s) requires the additional presence of other divalent metal ions, cobalt(III) hexamine or spermidine. Based on the suppression of Pb2+ cleavage at increasing concentrations of various divalent metal ions, our findings suggest that different divalent metal ions bind with different affinities to M1 RNA as well as to an RNase P hairpin–loop substrate and yeast tRNAPhe. We suggest that this approach can be used to obtain information about the relative binding strength for different divalent metal ions to RNA in general, as well as to specific RNA divalent metal ion binding sites. Of those studied in this report, Mn2+ is generally among the strongest RNA binders.

A mechanism for the inhibition of fever by a virus.

Poxviruses encode proteins that block the activity of cytokines. Here we show that the study of such virulence factors can contribute to our understanding of not only virus pathogenesis but also the physiological role of cytokines. Fever is a nonspecific response to infection that contributes to host defense. Several cytokines induce an elevation of body temperature when injected into animals, but in naturally occurring fever it has been difficult to show that any cytokine has a critical role. We describe the first example of the suppression of fever by a virus and the molecular mechanism leading to it. Several vaccinia virus strains including smallpox vaccines express soluble interleukin 1 (IL-1) receptors, which bind IL-1 beta but not IL-1 alpha. These viruses prevent the febrile response in infected mice, whereas strains that naturally or through genetic engineering lack the receptor induce fever. Repair of the defective IL-1 beta inhibitor in the smallpox vaccine Copenhagen, a more virulent virus than the widely used vaccine strains Wyeth and Lister, suppresses fever and attenuates the disease. The vaccinia-induced fever was inhibited with antibodies to IL-1 beta. These findings provide strong evidence that IL-1 beta, and not other cytokines, is the major endogenous pyrogen in a poxvirus infection.

The control of hematopoiesis and leukemia: from basic biology to the clinic.

Hematopoiesis gives rise to blood cells of different lineages throughout normal life. Abnormalities in this developmental program lead to blood cell diseases including leukemia. The establishment of a cell culture system for the clonal development of hematopoietic cells made it possible to discover proteins that regulate cell viability, multiplication and differentiation of different hematopoietic cell lineages, and the molecular basis of normal and abnormal blood cell development. These regulators include cytokines now called colony-stimulating factors (CSFs) and interleukins (ILs). There is a network of cytokine interactions, which has positive regulators such as CSFs and ILs and negative regulators such as transforming growth factor beta and tumor necrosis factor (TNF). This multigene cytokine network provides flexibility depending on which part of the network is activated and allows amplification of response to a particular stimulus. Malignancy can be suppressed in certain types of leukemic cells by inducing differentiation with cytokines that regulate normal hematopoiesis or with other compounds that use alternative differentiation pathways. This created the basis for the clinical use of differentiation therapy. The suppression of malignancy by inducing differentiation can bypass genetic abnormalities that give rise to malignancy. Different CSFs and ILs suppress programmed cell death (apoptosis) and induce cell multiplication and differentiation, and these processes of development are separately regulated. The same cytokines suppress apoptosis in normal and leukemic cells, including apoptosis induced by irradiation and cytotoxic cancer chemotherapeutic compounds. An excess of cytokines can increase leukemic cell resistance to cytotoxic therapy. The tumor suppressor gene wild-type p53 induces apoptosis that can also be suppressed by cytokines. The oncogene mutant p53 suppresses apoptosis. Hematopoietic cytokines such as granulocyte CSF are now used clinically to correct defects in hematopoiesis, including repair of chemotherapy-associated suppression of normal hematopoiesis in cancer patients, stimulation of normal granulocyte development in patients with infantile congenital agranulocytosis, and increase of hematopoietic precursors for blood cell transplantation. Treatments that decrease the level of apoptosis-suppressing cytokines and downregulate expression of mutant p53 and other apoptosis suppressing genes in cancer cells could improve cytotoxic cancer therapy. The basic studies on hematopoiesis and leukemia have thus provided new approaches to therapy.

Potentiation of the bioavailability of daidzin by an extract of Radix puerariae.

The dose effect of pure daidzin on the suppression of ethanol intake in Syrian golden hamsters was compared with that of crude daidzin contained in a methanol extract of Radix puerariae (RP). EC50 values estimated from the graded dose-response curves for pure daidzin and RP extract daidzin are 23 and 2.3 mg per hamster per day, respectively. Apparently the antidipsotropic activity of the RP extract cannot be accounted for solely by its daidzin content (22 mg/g). In addition to daidzin, six other isoflavones were identified in the RP extract and quantified--namely, puerarin (160 mg per g of extract), genistin (3.7 mg/g), daidzein (2.6 mg/g), daidzein-4',7-diglucoside (1.2 mg/g), genistein (0.2 mg/g), and formononetin (0.16 mg/g). None of these, administered either alone or combined, contributes in any significant way to the antidipsotropic activity of the extract. Plasma daidzin concentration-time curves determined in hamsters administered various doses of pure daidzin or RP extract by i.p.injection indicate that the crude extract daidzin has approximately 10 times greater bioavailability than the pure compound. Reconstruction of the dose-response effects for pure and crude daidzin using bioavailable daidzin rather than administered dose gives a single curve. Synthetic daidzin added to the RP extract acquires the bioavailability of the endogenous daidzin that exists naturally in the extract. These results show that (i) daidzin is the major active principle in methanol extracts of RP, and (ii) additional constituents in the methanol extract of RP assist uptake of daidzin in golden hamsters.