A synthetic biochemistry platform for cell free production of monoterpenes from glucose.

Cell-free systems designed to perform complex chemical conversions of biomass to biofuels or commodity chemicals are emerging as promising alternatives to the metabolic engineering of living cells. Here we design a system comprises 27 enzymes for the conversion of glucose into monoterpenes that generates both NAD(P)H and ATP in a modified glucose breakdown module and utilizes both cofactors for building terpenes. Different monoterpenes are produced in our system by changing the terpene synthase enzyme. The system is stable for the production of limonene, pinene and sabinene, and can operate continuously for at least 5 days from a single addition of glucose. We obtain conversion yields >95% and titres >15 g l-1. The titres are an order of magnitude over cellular toxicity limits and thus difficult to achieve using cell-based systems. Overall, these results highlight the potential of synthetic biochemistry approaches for producing bio-based chemicals

Computational Fluid Dynamic Optimization and Design for the Airborne Laser System

The Airborne Laser (ABL) was designed to destroy any ballistic missile shortly after launch that could be a threat to the United States and its allies. The ABL uses several lasers to accomplish the destruction of the ballistic missile, most notably the high powered Chemical Oxygen Iodine Laser (COIL). The COIL is a complex device that could be improved upon in several areas that will result in overall weight reduction, refinement of beam quality, and increased magazine capacity. This dissertation presents novel design and optimization techniques coupled with fluid dynamics to improve the performance of the COIL system. The focus was on two components of the COIL system: the iodine mixing nozzle and the pressure recovery system. Improvements to the iodine mixing nozzle were made in terms of mixing efficiency, gain uniformity, and flow uniformity. These improvements result in a power increase per module, which in turn reduces the overall number of modules required to shoot down a missile. The use of fewer modules significantly reduces the weight of the entire system. Additionally, investigations into the pressure recovery system led to further reduction in weight. New designs increased the mixing of the flows, which improved the pressure recovery and entrainment ratios. Focusing on the ABL application, the required pressure recovery needed for operation could be achieved with lower flow rates, and thus, less fluid is needed onboard

Gardenia (Rubiaceae) seed conservation physiology with emphasis on rare Hawaiian species

Background and aims – Gardenia species are ecologically, culturally, and economically significant but the three native species of Gardenia in Hawai‘i are assessed as Critically Endangered. Seed banking is the most cost effective and efficient means of conserving plant material ex situ. To better understand the conservation physiology of Hawaiian and South Pacific Gardenia spp. and support their conservation, we asked 1) How do seeds respond to different temperatures and light and dark regimes? 2) What class of dormancy, if any, do seeds exhibit? 3) How does seed germinability respond over time in a seed bank? and 4) What is the conservation status and level of ex situ representation of Gardenia globally? Material and methods – To answer these questions, we used 19 accessions of fresh seeds and seeds stored for varying periods of time in the National Tropical Botanical Garden’s Conservation Seed Bank and Laboratory of Hawaiian (G. brighamii, G. remyi), New Caledonian (G. aubryi, G. oudiepe), and Tahitian (G. taitensis) species. Seeds were incubated at varying temperatures and in light, and in dark. Key results – We found that (1) seeds of all species tested germinated slowly and only at higher temperatures in the light and dark, (2) seeds have non-deep physiological dormancy, (3) seeds of the Hawaiian species are short lived at conventional seed bank conditions, and (4) only 40% of Gardenia spp. are represented in ex situ facilities, and 66% of the species have not been evaluated for the IUCN Red List. Conclusion – Seeds of Hawaiian Gardenia spp. are short lived in storage. Since seeds germinate in darkness, they are unlikely to form a persistent soil seedbank. Although seeds of all species tested are physiologically dormant, they can be easily propagated from seed at warmer temperatures, giving some hope to the conservation and restoration of the Critically Endangered Hawaiian species. Since our dataset was limited by a lack of continuous viability monitoring, we emphasize the need for initial germination testing and ongoing viability tests to better understand seed longevity. Lastly, we discuss the ecological relevance of our results in the context of the Hawaiian archipelago

Bladder secretion of inhibitors of calcium oxalate crystal growth

Bladder secretion of inhibitors of calcium oxalate crystal growth.Differences in calcium oxalate crystal growth inhibition were studied in normally voided urine (bladder urine) and in urine collected directly from the kidney (kidney urine) in nine dogs. Urine samples were collected before and 10 days after bilateral ureterostomies. Calcium oxalate crystal growth inhibition was measured in a standard seeded crystal growth system. The alcian blue-precipitable material of the urine samples was determined. Significantly lower values were observed in kidney urine than in bladder urine for calcium oxalate crystal inhibition (mean difference, 0.07 ± 0.02 inhibitor units/mg creatinine; P < 0.01) and for the alcian blue-precipitable material (mean difference, 0.07 ± 0.02 mg/mg creatinine; P < 0.01). We conclude that the bladder adds calcium oxalate crystal growth inhibition to urine. Glycosaminoglycans from the bladder mucosa may be responsible; however, other acidic polymers such as RNA fragments or glycopeptides have been shown to be a constituent of the alcian blue-precipitable material. These are potent inhibitors of calcium oxalate crystal growth, and their participation in the increase of inhibition observed in bladder urine cannot be excluded. Total calcium oxalate crystal growth inhibition present in normally voided urine may be an overestimation of the actual inhibition present at the level of the kidney, where calculi usually form

NV FUEL OPTIMIZER

Trabajo Fin de Máster de Administración y Dirección de Empresas (MBA)con un proyecto de inversión NV FUEL OPTIMIZER, un proyecto conservador, ecológico y novedoso para el sector de la automoción, estratégico para la economía española, tanto por su elevado peso en la industria, el empleo y las exportaciones, así como por su efecto sobre el sector servicios. Con el objetivo de ofrecer un producto que ayude a hacer más kilómetros sin necesidad de cambiar de coche y ayude a cuidar el medio ambiente por una pequeña inversión rápidamente amortizable, definitivamente se constituye en una oportunidad de negocio

Orbits of crystallographic embedding of non-crystallographic groups and applications to virology

The architecture of infinite structures with non-crystallographic symmetries can be modelled via aperiodic tilings, but a systematic construction method for finite structures with non-crystallographic symmetry at different radial levels is still lacking. This paper presents a group theoretical method for the construction of finite nested point sets with non-crystallographic symmetry. Akin to the construction of quasicrystals, a non-crystallographic group G is embedded into the point group of a higher-dimensional lattice and the chains of all G-containing subgroups are constructed. The orbits of lattice points under such subgroups are determined, and it is shown that their projection into a lower-dimensional G-invariant subspace consists of nested point sets with G-symmetry at each radial level. The number of different radial levels is bounded by the index of G in the subgroup of . In the case of icosahedral symmetry, all subgroup chains are determined explicitly and it is illustrated that these point sets in projection provide blueprints that approximate the organization of simple viral capsids, encoding information on the structural organization of capsid proteins and the genomic material collectively, based on two case studies. Contrary to the affine extensions previously introduced, these orbits endow virus architecture with an underlying finite group structure, which lends itself better to the modelling of dynamic properties than its infinite-dimensional counterpart

Lessons from Two Design–Build–Test–Learn Cycles of Dodecanol Production in Escherichia coli Aided by Machine Learning

The Design–Build–Test–Learn (DBTL) cycle, facilitated by exponentially improving capabilities in synthetic biology, is an increasingly adopted metabolic engineering framework that represents a more systematic and efficient approach to strain development than historical efforts in biofuels and biobased products. Here, we report on implementation of two DBTL cycles to optimize 1-dodecanol production from glucose using 60 engineered Escherichia coli MG1655 strains. The first DBTL cycle employed a simple strategy to learn efficiently from a relatively small number of strains (36), wherein only the choice of ribosome-binding sites and an acyl-ACP/acyl-CoA reductase were modulated in a single pathway operon including genes encoding a thioesterase (UcFatB1), an acyl-ACP/acyl-CoA reductase (Maqu_2507, Maqu_2220, or Acr1), and an acyl-CoA synthetase (FadD). Measured variables included concentrations of dodecanol and all proteins in the engineered pathway. We used the data produced in the first DBTL cycle to train several machine-learning algorithms and to suggest protein profiles for the second DBTL cycle that would increase production. These strategies resulted in a 21% increase in dodecanol titer in Cycle 2 (up to 0.83 g/L, which is more than 6-fold greater than previously reported batch values for minimal medium). Beyond specific lessons learned about optimizing dodecanol titer in E. coli, this study had findings of broader relevance across synthetic biology applications, such as the importance of sequencing checks on plasmids in production strains as well as in cloning strains, and the critical need for more accurate protein expression predictive tools

Vaccinia virus lacking the Bcl-2-like protein N1 induces a stronger natural killer cell response to infection

The vaccinia virus (VACV) N1 protein is an intracellular virulence factor that has a Bcl-2-like structure and inhibits both apoptosis and signalling from the interleukin 1 receptor, leading to nuclear factor kappa B activation. Here, we investigated the immune response to intranasal infection with a virus lacking the N1L gene (vΔN1L) compared with control viruses expressing N1L. Data presented show that deletion of N1L did not affect the proportion of CD4+ and CD8+ T cells infiltrating the lungs or the cytotoxic T-cell activity of these cells. However, vΔN1L induced an increased local natural killer cell activity between days 4 and 6 post-infection. In addition, in the absence of N1 the host inflammatory infiltrate was characterized by a reduced proportion of lymphocytes bearing the early activation marker CD69. Notably, there was a good correlation between the level of CD69 expression and weight loss. The implications of these findings are discussed