Boundary layer thickness effect on boattail drag

A combined experimental and analytical program was conducted to investigate the effects of boundary layer changes on the flow over high angle boattail nozzles. The tests were run on an isolated axisymmetric sting mounted model. Various boattail geometries were investigated at high subsonic speeds over a range of boundary layer thicknesses. In general, boundary layer effects were small at speeds up to Mach 0.8. However, at higher speeds significant regions of separated flow were present on the boattail. When separation was present large reductions in boattail drag resulted with increasing boundary layer thickness. The analysis predicts both of these trends

Surpassing thermodynamic, kinetic, and stability barriers to isomerization catalysis for tagatose biosynthesis

D-Tagatose is a rare ketohexose sugar with sweetness similar to that of sucrose. However, its glycemic index and caloric value is much lower because of low bioavailability, making it an attractive GRAS (generally regarded as safe) sugar substitute. Recent studies have also demonstrated that it is anti-hyperglycemic and prebiotic, which promotes gut health. Thus, there exists a high demand in food industry for the economical production of rare sugars, like tagatose. The enzyme L-arabinose isomerase (LAI) that responsible for the reversible isomerization of the pentose L-arabinose to L-ribulose can also isomerize the hexose D-galactose to D-tagatose. LAI has thus been the enzyme of choice to produce tagatose, although, to date, few commercial bioprocesses exist. A variety of LAIs from different microorganisms have been isolated and have reported optimal activity at a range of temperatures and pH. Some of the limitations of tagatose biosynthesis using LAI that may be hindering commercial viability are, 1) unfavorable enzymatic kinetics since galactose is not the native substrate of LAI, 2) low enzyme stability, particularly in the absence of divalent metal ions, and 3) low equilibrium constant for galactose to tagatose isomerization. Few previous reports have been successful at engineering enzymatic properties of LAI for industrial application; often addressing only one of the bottlenecks to productivity. To address the kinetic issue, several groups have used enzyme engineering methods to enhance catalytic efficiency of LAI toward galactose and have shown moderate increases in productivity. To counter low-stability issues, many groups have tested the utility of thermophilic enzymes. However, most thermophilic enzymes rely on divalent metal ions (Mn2+, Co2+, Fe2+) for stability, and high reaction temperatures (≥ 80 °C) result in significant caramelization, which are all undesirable and must be removed from product, adding to processing costs. Surface-display or encapsulation in particles or whole-cells can stabilize enzymes. Finally, the thermodynamic limitations of isomerization of galactose to tagatose are severe and, arguably, the most recalcitrant issue since ΔG°rxn ≈ +1.2 kcal/mol, which indicates theoretical maximum equilibrium conversion ~ 14 % at room temperature. Several approaches have been used to overcome this limitation. Thermophilic enzymes can achieve higher conversions than mesophilic enzymes since the equilibrium shifts toward tagatose at higher temperatures. Whole-cell biocatalysts with GRAS organisms (e.g. lactic acid bacteria (LAB) and E. coli) that disproportionately partition substrate and product across their membrane has also been shown to partially circumvent this thermodynamic limitation while simultaneously enhancing enzyme stability; albeit at a kinetic penalty imposed by substrate transport limitations. Recently, cell permeabilization and sugar transport overexpression were demonstrated as methods to overcome the kinetic penalty imposed by cellular encapsulation. There have currently been no studies that look to systematically analyze all three limitations – kinetic, thermodynamic, and enzyme stability – of the enzymatic isomerization of galactose to tagatose. This work clearly demonstrates the presence of these three limitations and provides a novel approach to balance their advantages and limitations. We use the food-safe engineered probiotic bacterium Lactobacillus plantarum as the expression host due to its increasing relevance to biochemical and biomedical research. This approach enabled ~ 50 % conversion of galactose to tagatose in 4 h (productivity of ~ 38 mmol tagatose L-1 h-1) ultimately reaching ~ 85 % conversion after 48 h at high galactose loading (300 mM) in batch culture. This is among the highest conversions and productivities reported to date for tagatose production using a mesophilic enzyme. Such an approach is expected to be applicable to other biocatalytic systems where similar trade-offs between kinetics, thermodynamics, and/or stability pose hurdles to process development. This work is currently under consideration for publication: Bober & Nair (2019) Nature Communications (in revision) https://nature-research-under-consideration.nature.com/channels/1337-under-consideration/posts/44492-surpassing-thermodynamic-kinetic-and-stability-barriers-to-isomerization-catalysis-for-tagatose-biosynthesi

Strontium optical lattice clocks for practical realization of the metre and secondary representation of the second

We present a system of two independent strontium optical lattice standards probed with a single shared ultra-narrow laser. The absolute frequency of the clocks can be verified by the use of Er:fiber optical frequency comb with the GPS-disciplined Rb frequency standard. We report hertz-level spectroscopy of the clock line and measurements of frequency stability of the two strontium optical lattice clocks.Comment: This is an author-created, un-copyedited version of an article accepted for publication in Meas. Sci. Technol. The publisher is not responsible for any errors or omissions in this version of the manuscript or any version derived from it. The Version of Record is available online at doi:10.1088/0957-0233/26/7/07520

Hypothalamic Agrp Neurons Drive Stereotypic Behaviors beyond Feeding

SummaryThe nervous system evolved to coordinate flexible goal-directed behaviors by integrating interoceptive and sensory information. Hypothalamic Agrp neurons are known to be crucial for feeding behavior. Here, however, we show that these neurons also orchestrate other complex behaviors in adult mice. Activation of Agrp neurons in the absence of food triggers foraging and repetitive behaviors, which are reverted by food consumption. These stereotypic behaviors that are triggered by Agrp neurons are coupled with decreased anxiety. NPY5 receptor signaling is necessary to mediate the repetitive behaviors after Agrp neuron activation while having minor effects on feeding. Thus, we have unmasked a functional role for Agrp neurons in controlling repetitive behaviors mediated, at least in part, by neuropeptidergic signaling. The findings reveal a new set of behaviors coupled to the energy homeostasis circuit and suggest potential therapeutic avenues for diseases with stereotypic behaviors.PaperCli

Engineering the microbiota to treat metabolic disorders

Inborn errors of metabolism (IEM) are a family of more than 500 potentially lethal congenital genetic disorders that cumulatively affect 1 in 1000 newborns. In many IEMs, pathologies manifest as a result of improper metabolism of nutrients in food. In Phenylketonuria (PKU) for example, elevated levels of phenylalanine and the accumulation of aberrant metabolic intermediates in the system lead to acute and chronic toxicities. Resultantly, many disorders within this group are generally treated through lifelong nutritional management due to the lack of alternative and pharmacological options. Longitudinal studies have indicated that even with strict adherence to a diet of synthetic supplements, patients experience chronic issues like frailty, delayed growth, and intellectual disabilities. Recently, enzyme-replacement therapies (ERT) have demonstrated promise in pre-clinical and clinical settings by providing a metabolic sink for phenylalanine in PKU. As an enhancement to traditional ERT, we are developing a novel therapeutic for IEMs associated with amino acids by expressing metabolic enzymes in lactic acid bacteria (LAB) that natively colonize the human gastrointestinal (GI) tract. Starting with an enzyme under clinical development for PKU, phenylalanine ammonia-lyase (PAL), and by promoting the intestinal adhesion and colonization characteristics, the engineered LAB will intervene before amino acid absorption occurs in the small intestines during digestion. To engineer new enzymes with activities required for treating IEMs, we have developed a novel facile selection and screening methodology. This can potentially be utilized to enhance enzymatic properties or identify mutants with altered substrate specificity, creating a spectrum of PALs that can be used to treat IEMs associated with other amino acids. Here we describe the methodology, development, and optimization of this method. To characterize and engineer microbial adhesion to intestinal mucus, we developed a novel assay that is able to capture the quantitative and mechanistic binding thermodynamics of cells to mucus. We will discuss the development of this assay and its implementation for engineering improved mucus binding. The platform technologies discussed here will be instrumental in realizing microbiota-based therapeutics as an emerging and urgently-needed treatment for IEMs that currently have inadequate or no options

Zeeman slowers made simple with permanent magnets in a Halbach configuration

We describe a simple Zeeman slower design using permanent magnets. Contrary to common wire-wound setups no electric power and water cooling are required. In addition, the whole system can be assembled and disassembled at will. The magnetic field is however transverse to the atomic motion and an extra repumper laser is necessary. A Halbach configuration of the magnets produces a high quality magnetic field and no further adjustment is needed. After optimization of the laser parameters, the apparatus produces an intense beam of slow and cold 87Rb atoms. With a typical flux of 1 - 5 \times 10^10 atoms/s at 30 ms^-1, our apparatus efficiently loads a large magneto-optical trap with more than 10^10 atoms in one second, which is an ideal starting point for degenerate quantum gases experiments.Comment: 8+6 pages (article + appendices: calculation details, probe and oven description, pictures), 18 figures, supplementary material (movie, Mathematica programs and technical drawings

A conjectural extension of Hecke’s converse theorem

We formulate a precise conjecture that, if true, extends the converse theorem of Hecke without requiring hypotheses on twists by Dirichlet characters or an Euler product. The main idea is to linearize the Euler product, replacing it by twists by Ramanujan sums. We provide evidence for the conjecture, including proofs of some special cases and under various additional hypotheses

Fibre-optic delivery of time and frequency to VLBI station

The quality of Very Long Baseline Interferometry (VLBI) radio observations predominantly relies on precise and ultra-stable time and frequency (T&F) standards, usually hydrogen masers (HM), maintained locally at each VLBI station. Here, we present an operational solution in which the VLBI observations are routinely carried out without use of a local HM, but using remote synchronization via a stabilized, long-distance fibre-optic link. The T&F reference signals, traceable to international atomic timescale (TAI), are delivered to the VLBI station from a dedicated timekeeping laboratory. Moreover, we describe a proof-of-concept experiment where the VLBI station is synchronized to a remote strontium optical lattice clock during the observation.Comment: 8 pages, 8 figures, matches the version published in A&A, section Astronomical instrumentatio