1,907,353 research outputs found
Optimization of Pseudomonas putida KT2440 as host for the production of cis, cis-muconate from benzoate
No full text Optimization of Pseudomonas putida KT2440 as host for the production of cis, cis-muconate from benzoate P. putida KT2440 was used as biocatalyst given its versatile and energetically robust metabolism. Therefore, a mutant was generated and a process developed based on which a life cycle assessment (LCA) was performed. Additionally, the growth related parameters were experimentally obtained to constrain the metabolic model iJP815 further. The mutant Pseudomonas putida KT2440-JD1 was derived from P. putida KT2440 after NTGmutagenesis and exposure to 3-fluorobenzoate. The strain was no longer able to grow with benzoate as a single source of carbon and energy. Instead, benzoate was co-metabolized to cis, cis-muconate that accumulated in the culture medium while the strain grew on glucose. In batch cultures, a maximal production rate per gram biomass of 2.0 g cis, cis-muconate gDCW -1 h-1 was obtained. This is 8-fold higher than thus far reported. The cat operon was no longer expressed in P. putida KT2440-JD1 due to a point mutation in the regulator gene catR. This operon contains the genes for the conversion of catechol to metabolites of the central metabolism, including catA, which encodes a catechol 1,2-dioxygenase. Consequently, benzoate is converted in the mutant by enzymes that are encoded on the ben operon. This operon includes a gene (catA2; PP_3166) that encodes an additional catechol 1,2-dioxygenase, thus allowing the conversion of benzoate to cis, cis-muconate. In batch cultures the maximal growth rate of P. putida KT2440-JD1 in mineral medium with glucose decreased linearly in the presence of increasing concentrations of benzoate and/or cis, cismuconate and finally stopped at 6 g L-1 benzoate or 85 g L-1 cis, cis-muconate. The inhibitory effects of both compounds were cumulative and no synergistic effects were observed. The maximal uptake rate of benzoate was higher than the production rate of cis, cis-muconate per gram biomass during growth on glucose in the presence of benzoate, indicating that a benzoate derivative accumulated in the cells, which is likely to be catechol, as accumulation of this intermediate was observed. Catechol is known to cause oxidative stress, and the accumulation of catechol and benzoate should be prevented during the production of cis, cis-muconate. This is feasible by coupling the addition of benzoate to the decrease of the pH of the culture medium in a so-called pHstat process, as the cultivation medium acidifies only when benzoate has been converted to cis, cismuconate. Such a pH-stat fed-batch process resulted in the production of 18.5 g L-1 cis, cismuconate from benzoate with a molar yield of 96%. The phenotype of P. putida KT2440 can be assessed by the constraint-based metabolic model iJP815. The solution space of the model based on flux variability analysis was further constrained by growth associated maintenance, non-growth associated maintenance, and biomass composition determined form the experimentally measured growth-related factors of the strain that were generated during continuous cultivations at various dilution rates (D) (0.05-0.49) h-1. Transcriptomic profiles obtained at a D of 0.2 h-1 were consistent with model predictions of expressed genes based on flux balance analysis. The growth-related macro molecular composition of the biomass was similar as measured with E. coli K-12. However, growth parameters like the maximum biomass yield and maintenance coefficient were different. The energy required for assembly of P. putida KT2440 is higher compared to E. coli W3110, which will result in higher costs of glucose in biotechnological processes. On the other hand, the metabolism is robust as even at a D of 0.49 h-1 no overflow metabolism was observed. The lack of overflow metabolism is of great importance as it underscores the capacity of P. putida KT2440 as biocatalyst e.g. for conversions involving cofactor dependent oxygenase reactions. Further restriction of enzymatic conversions with already published 13C measurements only decreased the maximum solution space excluding solutions that are far from the experimental phenotype. Possibly, the best agreement was reached within limitations of constraint-based modeling. A LCA was performed on a combined biological and chemical process for the production of adipic acid. The outcome was compared with the traditional chemical process. The LCA focused on the cumulative energy demand, cumulative exergy demand and the CO2 equivalent emissions, with CO2 and N2O separate. Acidified cis, cis-muconate can be easily hydrogenated to adipic acid a resource for nylon-6,6 used in carpets and the auto industry, because of its long lasting and strong features. Feedstocks have a large effect on the overall environmental impact. The soil bacterium P. putida KT2440 has a versatile metabolism and is able to convert various feedstocks to cis, cis-muconate. Consequently, the use of feedstocks with a lower energy demand were taken into account besides benzoate, including: the petrochemical based feedstocks impure aromatics and toluene, and the biomass based feedstock phenol (from lignin). The effect of an increase of the final concentration cis, cis-muconate in the fermentor broth from 1.85% to 4.26% was modeled as P. putida KT2440- JD1 was able to consume benzoate up to 4.26% cis, cis-muconate. At a final concentration of 1.85% cis, cis-muconate, the use of impure aromatics and lignin instead of benzoate reduced the energy demand compared to the chemical production of adipic acid. The applicability of these feedstocks depends on the metabolic robustness of P. putida KT2440-JD1 with impure aromatics, and/or the development of an efficient process for the production of phenol from lignin. At a final concentration of 4.26% cis, cis-muconate the process energy and CO2eq emissions were reduced for all feedstocks. The pH-stat fed-batch process had a higher production rate per gram biomass compared to other processes although the speed was significantly lower as measured during the batch cultures. By generating a high cell density in combination of a pH-regulated inflow of benzoate, the volumetric productivity will increase as has been described for a cell-recycle process. While this only has a minor impact on the energy consumption, it does have a major impact on the economic parameters of an industrial scale bio-reactor. Recycling biomass with a membrane and using a solvent, as e.g. cold diethyl ether, after the acidification of the medium (pH 2.5) for the extraction of cis, cismuconate, could further reduce the environmental impact. The use of solvents will result in a higher product concentration and/or the evaporation will require less energy since the boiling temperature of solvents is most often lower than water. </p
CIS-7 Perspective on Trade with EU in the Context of EU Enlargement
No full textThe paper analyses possible consequences of the EU enlargement on the EU – CIS-7 trade. It considers current situation in trade between two country groups, describes the factors limiting this trade, and discusses the opportunities for the trade associated with the EU accession of the Central and East European countries with strong historical ties to the CIS-7. The paper concludes that the EU enlargement creates some potential for expansion of trade and, importantly, exports from CIS-7 to Europe.EU enlargement, CIS-7, foreign trade
GluK2-cis
No full textThe subdirectory /MD includes five free MD simulation trajectories of GluK2-cis complex described in the manuscript. The subdirectory /isomerization includes five forced switching simulation trajectories from cis-gluazo to trans-gluazo. The subdirectory /umbrella-clamshell includes the files needed to reproduce the umbrella sampling simulation results of LBD clamshell motions of GluK2-cis complex. This data file could be used to reproduce all the results of GluK2-cis complex presented in the manuscript
Policy Challenges Faced by Low-Income CIS Economies
No full textIn the 1990s, the CIS region experienced a painful transformation following the collapse of the USSR and the command economy. For the less developed republics of the former USSR, this process was even more dramatic as they lost subsidies from the Union's budget and some of them suffered devastating conflicts. In the 2000s, after overcoming the adaptation output decline and the consequences of the 1998-1999 financial crises, these economies started to grow rapidly, reducing poverty and macroeconomic imbalances. However, their future growth prospects are increasingly vulnerable due to their strong dependence on commodity exports, a poor business and investment climate, endemic corruption and weak governance. Quite recently, fighting high inflation has returned to the policy agenda. The modernization and diversification of the low-income CIS economies requires further market and institutional reforms aimed at overcoming the Soviet legacy of a repressive and inefficient state. The international community can help by resolving regional conflicts, assisting with trade and economic integration, and offering well-targeted development assistance.Commonwealth of Independent States, transition, low income countries,Southern Caucasus, Central Asia, trade, investment climate
A multiple-instance scoring method to predict tissue-specific cis-regulatory motifs and regions
No full textTranscription is the central process of gene regulation. In higher eukaryotes, the transcription of a gene is usually regulated by multiple cis-regulatory regions (CRRs). In different tissues, different transcription factors bind to their cis-regulatory motifs in these CRRs to drive tissue-specific expression patterns of their target genes. By combining the genome-wide gene expression data with the genomic sequence data, we proposed multiple-instance scoring (MIS) method to predict the tissue-specific motifs and the corresponding CRRs. The method is mainly based on the assumption that only a subset of CRRs of the expressed gene should function in the studied tissue. By testing on the simulated datasets and the fly muscle dataset, MIS can identify true motifs when noise is high and shows higher specificity for predicting the tissue-specific functions of CRRs
Cis-hetero-misogyny online
No full textThis article identifies five genres of anti-queer hate speech found in The Australian’s Facebook comments sections, exposing and analyzing the ways in which such comments are used to derogate cisgender and (often) heterosexual women. One may be tempted to think of cis-het women as third-party victims of queerphobia; however, this article argues that these genres of anti-queer speech are, in fact, misogynistic. Specifically, it argues that these are instances of cis-hetero-misogynistic hate speech. Cis-hetero-misogyny functions as the “law enforcement branch” of a cis-hetero-patriarchal gender order. Given the existence of such an order, it is clear that cis-het women’s liberation is inextricable from queer liberation (and vice versa). This article argues that to facilitate allyship and challenge this gender order—the order that elicits such hate speech acts—we need an epistemological revolution in the way we recognize and re-cognize human difference
cis: v1.3.1
No full text<a class="anchor" href="#cis-131-updates"><span class="octicon octicon-link"></span></a>CIS 1.3.1 updates
<ul>
<li>JASCIS-231 & JASCIS-209 - CIS now better determines the yaxis when the user specifies the xaxis as 'time' so that overlaying multiple time series is easy</li>
<li>JASCIS-283 - An issue with setting xmin or xmax using datetimes</li>
<li>A minor fix to the AerosolCCI product</li>
</ul>
Cytokine-induced Src homology 2 protein (CIS) promotes T cell receptor-mediated proliferation and prolongs survival of activated T cells
No full textCopyright @ 2000 The Rockefeller University Press. This article is shared under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.Members of the suppressor of cytokine signaling (SOCS) family were discovered as negative regulators of cytokine signaling by inhibition of the Janus kinase–signal transducer and activator of transcription (Jak-STAT) pathway. Among them, cytokine-induced Src homology 2 (SH2) protein (CIS) was found to inhibit the interleukin 3– and erythropietin-mediated STAT5 signaling pathway. However, involvement of SOCS proteins in other signaling pathways is still unknown. This study shows that the expression of CIS is selectively induced in T cells after T cell receptor (TCR) stimulation. In transgenic mice, with selective expression of CIS in CD4 T cells, elevated CIS strongly promotes TCR-mediated proliferation and cytokine production in vitro, and superantigen-induced T cell activation in vivo. Forced expression of CIS also prolongs survival of CD4 T cells after TCR activation. Molecular events immediately downstream from the TCR are not changed in CIS-expressing CD4 T cells, but activation of mitogen-activated protein (MAP) kinase pathways by TCR stimulation is significantly enhanced. Together with the increased MAP kinase activation, a direct interaction of CIS and protein kinase Cθ was also demonstrated. These results suggest that CIS is one of the important regulators of TCR-mediated T cell activation. The functions of CIS, enhancing TCR signaling and inhibiting cytokine signaling, may be important in the regulation of immune response and homeostasis.The Swedish Medical Council, the Foundation for Strategic Research
(SSF) and the Medical faculty at Lund University
Las deudas pendientes de Latinoamérica.
No full textLa décima versión de la Cátedra Alberto Hurtado de Liderazgo Social, organizada por el
Centro de Reflexión y Acción Social (CREAS) de la Universidad Alberto Hurtado en conjunto
con el Centro de Investigación Social (CIS) de Un Techo para Chile, se llevó a cabo
entre el 21 de agosto y el 9 de octubre de 2007. En esta versión, titulada “Las deudas
pendientes de Latinoamérica” se estableció un espacio de reflexión y aprendizaje sobre
la realidad de las sociedades de los distintos países que componen América Latina. El
curso convocó a expertos de la escena política, académica y social que dialogaron con
estudiantes de educación superior y debatieron frente a las preguntas ¿cuáles son las
deudas pendientes en el continente? y ¿qué podemos hacer para subsanarlas
- …
