SYNZIP Protein Interaction Toolbox: in Vitro and in Vivo Specifications of Heterospecific Coiled-Coil Interaction Domains

The synthetic biology toolkit contains a growing number of parts for regulating transcription and translation, but very few that can be used to control protein association. Here we report characterization of 22 previously published heterospecific synthetic coiled-coil peptides called SYNZIPs. We present biophysical analysis of the oligomerization states, helix orientations, and affinities of 27 SYNZIP pairs. SYNZIP pairs were also tested for interaction in two cell-based assays. In a yeast two-hybrid screen, >85% of 253 comparable interactions were consistent with prior in vitro measurements made using coiled-coil microarrays. In a yeast-signaling assay controlled by coiled-coil mediated scaffolding, 12 SYNZIP pairs were successfully used to down-regulate the expression of a reporter gene following treatment with α-factor. Characterization of these interaction modules dramatically increases the number of available protein interaction parts for synthetic biology and should facilitate a wide range of molecular engineering applications. Summary characteristics of 27 SYNZIP peptide pairs are reported in specification sheets available in the Supporting Information and at the SYNZIP Web site [http://keatingweb.mit.edu/SYNZIP/].National Science Foundation (U.S.) (NSF award MCB 0950233)National Institutes of Health (U.S.) (grant RO1 GM55040)National Institutes of Health (U.S.) (grant PN2 EY016546)National Institutes of Health (U.S.) (grant P50 GMO81879)National Science Foundation (U.S.). Synthetic Biology Engineering Research CenterHoward Hughes Medical Institut

A Synthetic Coiled-Coil Interactome Provides Heterospecific Modules for Molecular Engineering

The versatile coiled-coil protein motif is widely used to induce and control macromolecular interactions in biology and materials science. Yet the types of interaction patterns that can be constructed using known coiled coils are limited. Here we greatly expand the coiled-coil toolkit by measuring the complete pairwise interactions of 48 synthetic coiled coils and 7 human bZIP coiled coils using peptide microarrays. The resulting 55-member protein “interactome” includes 27 pairs of interacting peptides that preferentially heteroassociate. The 27 pairs can be used in combinations to assemble sets of 3 to 6 proteins that compose networks of varying topologies. Of special interest are heterospecific peptide pairs that participate in mutually orthogonal interactions. Such pairs provide the opportunity to dimerize two separate molecular systems without undesired crosstalk. Solution and structural characterization of two such sets of orthogonal heterodimers provide details of their interaction geometries. The orthogonal pair, along with the many other network motifs discovered in our screen, provide new capabilities for synthetic biology and other applications.National Institutes of Health (U.S.) (NIH Award GM067681)National Institutes of Health (U.S.) (NCRR Award RR-15301

3-Bromophenyl 6-acetoxymethyl-2-oxo-2H-1-benzopyran-3-carboxylate inhibits cancer cell invasion in vitro and tumour growth in vivo

In search for new anticancer agents, we have evaluated the antiinvasive and antimigrative properties of recently developed synthetic coumarin derivatives among which two compounds revealed important activity: 3-chlorophenyl 6-acetoxymethyl-2-oxo-2H-1-benzopyran-3-carboxylate and 3-bromophenyl 6-acetoxymethyl-2-oxo-2H-1-benzopyran-3-carboxylate, Both drugs were able to inhibit cell invasion markedly in a Boyden chamber assay, the bromo derivative being more potent than the reference matrix metalloprotease (MMP) inhibitor GI 129471. In vivo, tumour growth was reduced when nude mice grafted with HT 1080 or MDA-MB231 cells were treated i.p. 3 days week(-1) with the bromo coumarin derivative. These effects were not associated with the inhibition of urokinase, plasmin, MMP-2 or MMP-9. The mechanism of action of the drugs remains to be elucidated. However, these two coumarin derivatives may serve as new lead compounds of an original class of antitumour agents

Synthetic Biology: Mapping the Scientific Landscape

This article uses data from Thomson Reuters Web of Science to map and analyse the scientific landscape for synthetic biology. The article draws on recent advances in data visualisation and analytics with the aim of informing upcoming international policy debates on the governance of synthetic biology by the Subsidiary Body on Scientific, Technical and Technological Advice (SBSTTA) of the United Nations Convention on Biological Diversity. We use mapping techniques to identify how synthetic biology can best be understood and the range of institutions, researchers and funding agencies involved. Debates under the Convention are likely to focus on a possible moratorium on the field release of synthetic organisms, cells or genomes. Based on the empirical evidence we propose that guidance could be provided to funding agencies to respect the letter and spirit of the Convention on Biological Diversity in making research investments. Building on the recommendations of the United States Presidential Commission for the Study of Bioethical Issues we demonstrate that it is possible to promote independent and transparent monitoring of developments in synthetic biology using modern information tools. In particular, public and policy understanding and engagement with synthetic biology can be enhanced through the use of online interactive tools. As a step forward in this process we make existing data on the scientific literature on synthetic biology available in an online interactive workbook so that researchers, policy makers and civil society can explore the data and draw conclusions for themselves

Erratum to: 36th International Symposium on Intensive Care and Emergency Medicine

[This corrects the article DOI: 10.1186/s13054-016-1208-6.]

Global patient outcomes after elective surgery: prospective cohort study in 27 low-, middle- and high-income countries.

BACKGROUND: As global initiatives increase patient access to surgical treatments, there remains a need to understand the adverse effects of surgery and define appropriate levels of perioperative care. METHODS: We designed a prospective international 7-day cohort study of outcomes following elective adult inpatient surgery in 27 countries. The primary outcome was in-hospital complications. Secondary outcomes were death following a complication (failure to rescue) and death in hospital. Process measures were admission to critical care immediately after surgery or to treat a complication and duration of hospital stay. A single definition of critical care was used for all countries. RESULTS: A total of 474 hospitals in 19 high-, 7 middle- and 1 low-income country were included in the primary analysis. Data included 44 814 patients with a median hospital stay of 4 (range 2-7) days. A total of 7508 patients (16.8%) developed one or more postoperative complication and 207 died (0.5%). The overall mortality among patients who developed complications was 2.8%. Mortality following complications ranged from 2.4% for pulmonary embolism to 43.9% for cardiac arrest. A total of 4360 (9.7%) patients were admitted to a critical care unit as routine immediately after surgery, of whom 2198 (50.4%) developed a complication, with 105 (2.4%) deaths. A total of 1233 patients (16.4%) were admitted to a critical care unit to treat complications, with 119 (9.7%) deaths. Despite lower baseline risk, outcomes were similar in low- and middle-income compared with high-income countries. CONCLUSIONS: Poor patient outcomes are common after inpatient surgery. Global initiatives to increase access to surgical treatments should also address the need for safe perioperative care. STUDY REGISTRATION: ISRCTN5181700

Modeling the ocean and atmosphere during an extreme bora event in northern Adriatic using one-way and two-way atmosphere–ocean coupling

We have studied the performances of (a) a two-way coupled atmosphere–ocean modeling system and (b) one-way coupled ocean model (forced by the atmosphere model), as compared to the available in situ measurements during and after a strong Adriatic bora wind event in February 2012, which led to extreme air–sea interactions. The simulations span the period between January and March 2012. The models used were ALADIN (Aire Limitée Adaptation dynamique Développement InterNational) (4.4 km resolution) on the atmosphere side and an Adriatic setup of Princeton ocean model (POM) (1°∕30 × 1°∕30 angular resolution) on the ocean side. The atmosphere–ocean coupling was implemented using the OASIS3-MCT model coupling toolkit. Two-way coupling ocean feedback to the atmosphere is limited to sea surface temperature. We have compared modeled atmosphere–ocean fluxes and sea temperatures from both setups to platform and CTD (conductivity, temperature, and depth) measurements from three locations in the northern Adriatic. We present objective verification of 2 m atmosphere temperature forecasts using mean bias and standard deviation of errors scores from 23 meteorological stations in the eastern part of Italy. We show that turbulent fluxes from both setups differ up to 20 % during the bora but not significantly before and after the event. When compared to observations, two-way coupling ocean temperatures exhibit a 4 times lower root mean square error (RMSE) than those from one-way coupled system. Two-way coupling improves sensible heat fluxes at all stations but does not improve latent heat loss. The spatial average of the two-way coupled atmosphere component is up to 0.3 °C colder than the one-way coupled setup, which is an improvement for prognostic lead times up to 20 h. Daily spatial average of the standard deviation of air temperature errors shows 0.15 °C improvement in the case of coupled system compared to the uncoupled. Coupled and uncoupled circulations in the northern Adriatic are predominantly wind-driven and show no significant mesoscale differences

Modeling the ocean and atmosphere during an extreme bora event in northern Adriatic using one-way and two-way atmosphere-ocean coupling

We have studied the performances of (a) a two-way coupled atmosphere-ocean modeling system and (b) one-way coupled ocean model (forced by the atmosphere model), as compared to the available in situ measurements during and after a strong Adriatic bora wind event in February 2012, which led to extreme air-sea interactions. The simulations span the period between January and March 2012. The models used were ALADIN (Aire Limiteé Adaptation dynamique Développement InterNational) (4.4 km resolution) on the atmosphere side and an Adriatic setup of Princeton ocean model (POM) (130 × 130 angular resolution) on the ocean side. The atmosphere-ocean coupling was implemented using the OASIS3-MCT model coupling toolkit. Two-way coupling ocean feedback to the atmosphere is limited to sea surface temperature. We have compared modeled atmosphere-ocean fluxes and sea temperatures from both setups to platform and CTD (conductivity, temperature, and depth) measurements from three locations in the northern Adriatic. We present objective verification of 2 m atmosphere temperature forecasts using mean bias and standard deviation of errors scores from 23 meteorological stations in the eastern part of Italy. We show that turbulent fluxes from both setups differ up to 20 % during the bora but not significantly before and after the event. When compared to observations, two-way coupling ocean temperatures exhibit a 4 times lower root mean square error (RMSE) than those from one-way coupled system. Two-way coupling improves sensible heat fluxes at all stations but does not improve latent heat loss. The spatial average of the two-way coupled atmosphere component is up to 0.3 °C colder than the one-way coupled setup, which is an improvement for prognostic lead times up to 20 h. Daily spatial average of the standard deviation of air temperature errors shows 0.15 °C improvement in the case of coupled system compared to the uncoupled. Coupled and uncoupled circulations in the northern Adriatic are predominantly wind-driven and show no significant mesoscale differences. © 2016 Author(s)