Urban Egyptian Women Aged 19-30 Years Display Nutrition Transition-Like Dietary Patterns, with High Energy and Sodium Intakes, and Insufficient Iron, Vitamin D, and Folate Intakes.

BACKGROUND: Recent changes in Egyptian dietary habits can be attributed to more urban and sedentary lifestyles and to alterations in the dietary and economic context. The mean BMI of Egyptian women is one of the highest worldwide, and 50% have iron deficiency. OBJECTIVE: The aim was to quantify food and nutrient intakes of urban Egyptian women and conduct a detailed analysis of micronutrients commonly consumed in inadequate amounts, such as iron, vitamin D, and folate. METHODS: Urban Egyptian women aged 19-30 y (n = 130) were recruited during 2016-2017. Energy needs were estimated using the Henry equation, assuming a low physical activity level (1.4). Dietary intakes and iron bioavailability were estimated from a 4-d food diary. Macronutrient intakes were compared with WHO/FAO population goals and micronutrient intakes with Egyptian recommendations. Iron needs were determined for each subject. RESULTS: The mean BMI (kg/m2) was 27.9 ± 4.9. The mean total energy intake (TEI; 2389 ± 715 kcal/d) was significantly higher than needs (2135 ± 237 kcal/d; P = 0.00018). Total fat (33%TEI) and SFA (11%TEI) intakes were slightly higher than population goals (15-30%TEI and <10%TEI, respectively). Diets provided 18 ± 8 g/d of fiber, 98 ± 54 g/d of total sugars, and nearly twice the recommended sodium intake (intake: 2787 ± 1065 mg/d; recommendation: <1500 mg/d). Estimated dietary iron bioavailability was low (9.2% ± 1.6%), and 79% of women consumed less iron than the average requirement (17.5 ± 7 mg/d). Overall, 82% and 80% of women consumed less vitamin D and folate, respectively, than recommended. CONCLUSIONS: Egyptian women aged 19-30 y have high intakes of energy and sodium, whereas iron, vitamin D, and folate intakes are insufficient, with only low concentrations of bioavailable iron. These results call for further investigation into measures that would improve this population's diet quality.Publication from previous employment rol

Using high-throughput barcode sequencing to efficiently map connectomes

The function of a neural circuit is determined by the details of its synaptic connections. At present, the only available method for determining a neural wiring diagram with single synapse precision-a 'connectome'-is based on imaging methods that are slow, labor-intensive and expensive. Here, we present SYNseq, a method for converting the connectome into a form that can exploit the speed and low cost of modern high-throughput DNA sequencing. In SYNseq, each neuron is labeled with a unique random nucleotide sequence-an RNA 'barcode'-which is targeted to the synapse using engineered proteins. Barcodes in pre- and postsynaptic neurons are then associated through protein-protein crosslinking across the synapse, extracted from the tissue, and joined into a form suitable for sequencing. Although our failure to develop an efficient barcode joining scheme precludes the widespread application of this approach, we expect that with further development SYNseq will enable tracing of complex circuits at high speed and low cost

Prenatal Ultrasound Screening: False Positive Soft Markers May Alter Maternal Representations and Mother-Infant Interaction

International audienceBackground : In up to 5% of pregnancies, ultrasounds screenin detects a " soft marker" (SM) that places the foetus at risk for a severe abnormality. In most cases, prenatal diagnostic work-up rules out a severe defect. We aimed to study the effects of false positive SM on maternal emotional status, maternal representations of the infant, and mother-infant interaction. Methology and Principal Findings : Utilizing an extreme-case prospective case control design, we selected frome a group of 244 women undergroing ultrasound, 19 pregnant women whose foetus had a positive SM screening and reassuring diagnostic work up, and 19 controls without SM matched for age and education. In the third trimester of pregnancy, within one week after delivery, and 2 months postpartum, we assessed anxiety, depression and maternal representations; Mother-infant interactions were videotaped during feeding within one week after delivery and again at 2 months postpartum and coded blindly using Coding Interactive behavior (CIB) scales. Anxiety and depression scores were significantly higher at alla ssessment points in the SM group. Maternal representations were also observd in the SM groyp. These dyads showed greater dysregulation, lower maternal sensitivity, higher maternal intrusive behaviour and higher infant avoidance. Multivariate analysis showed that maternal representation and depression at third trimester predicted mother-infant interaction.Conclusion : False positive ultrasound screenings for SM are not benign and negatively affect the developping maternal-infant attachment. Medical efforts should be directed to minimize as much as possible such false diagnoses, and to the psychological adverse consequences

Copolymerization of single-cell nucleic acids into balls of acrylamide gel

We show the use of 5'-Acrydite oligonucleotides to copolymerize single-cell DNA or RNA into balls of acrylamide gel (BAGs). Combining this step with split-and-pool techniques for creating barcodes yields a method with advantages in cost and scalability, depth of coverage, ease of operation, minimal cross-contamination, and efficient use of samples. We perform DNA copy number profiling on mixtures of cell lines, nuclei from frozen prostate tumors, and biopsy washes. As applied to RNA, the method has high capture efficiency of transcripts and sufficient consistency to clearly distinguish the expression patterns of cell lines and individual nuclei from neurons dissected from the mouse brain. By using varietal tags (UMIs) to achieve sequence error correction, we show extremely low levels of cross-contamination by tracking source-specific SNVs. The method is readily modifiable, and we will discuss its adaptability and diverse applications

The synthetic integron: an in vivo genetic shuffling device

As the field of synthetic biology expands, strategies and tools for the rapid construction of new biochemical pathways will become increasingly valuable. Purely rational design of complex biological pathways is inherently limited by the current state of our knowledge. Selection of optimal arrangements of genetic elements from randomized libraries may well be a useful approach for successful engineering. Here, we propose the construction and optimization of metabolic pathways using the inherent gene shuffling activity of a natural bacterial site-specific recombination system, the integron. As a proof of principle, we constructed and optimized a functional tryptophan biosynthetic operon in Escherichia coli. The trpA-E genes along with ‘regulatory’ elements were delivered as individual recombination cassettes in a synthetic integron platform. Integrase-mediated recombination generated thousands of genetic combinations overnight. We were able to isolate a large number of arrangements displaying varying fitness and tryptophan production capacities. Several assemblages required as many as six recombination events and produced as much as 11-fold more tryptophan than the natural gene order in the same context

Development of an In Vitro Compartmentalization Screen for High-Throughput Directed Evolution of [FeFe] Hydrogenases

BACKGROUND: [FeFe] hydrogenase enzymes catalyze the formation and dissociation of molecular hydrogen with the help of a complex prosthetic group composed of common elements. The development of energy conversion technologies based on these renewable catalysts has been hindered by their extreme oxygen sensitivity. Attempts to improve the enzymes by directed evolution have failed for want of a screening platform capable of throughputs high enough to adequately sample heavily mutated DNA libraries. In vitro compartmentalization (IVC) is a powerful method capable of screening for multiple-turnover enzymatic activity at very high throughputs. Recent advances have allowed [FeFe] hydrogenases to be expressed and activated in the cell-free protein synthesis reactions on which IVC is based; however, IVC is a demanding technique with which many enzymes have proven incompatible. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe an extremely high-throughput IVC screen for oxygen-tolerant [FeFe] hydrogenases. We demonstrate that the [FeFe] hydrogenase CpI can be expressed and activated within emulsion droplets, and identify a fluorogenic substrate that links activity after oxygen exposure to the generation of a fluorescent signal. We present a screening protocol in which attachment of mutant genes and the proteins they encode to the surfaces of microbeads is followed by three separate emulsion steps for amplification, expression, and evaluation of hydrogenase mutants. We show that beads displaying active hydrogenase can be isolated by fluorescence-activated cell-sorting, and we use the method to enrich such beads from a mock library. CONCLUSIONS/SIGNIFICANCE: [FeFe] hydrogenases are the most complex enzymes to be produced by cell-free protein synthesis, and the most challenging targets to which IVC has yet been applied. The technique described here is an enabling step towards the development of biocatalysts for a biological hydrogen economy

Microfluidic Technologies for Synthetic Biology

Microfluidic technologies have shown powerful abilities for reducing cost, time, and labor, and at the same time, for increasing accuracy, throughput, and performance in the analysis of biological and biochemical samples compared with the conventional, macroscale instruments. Synthetic biology is an emerging field of biology and has drawn much attraction due to its potential to create novel, functional biological parts and systems for special purposes. Since it is believed that the development of synthetic biology can be accelerated through the use of microfluidic technology, in this review work we focus our discussion on the latest microfluidic technologies that can provide unprecedented means in synthetic biology for dynamic profiling of gene expression/regulation with high resolution, highly sensitive on-chip and off-chip detection of metabolites, and whole-cell analysis

Enhancing Protease Activity Assay in Droplet-Based Microfluidics Using a Biomolecule Concentrator

We introduce an integrated microfluidic device consisting of a biomolecule concentrator and a microdroplet generator, which enhances the limited sensitivity of low-abundance enzyme assays by concentrating biomolecules before encapsulating them into droplet microreactors. We used this platform to detect ultralow levels of matrix metalloproteinases (MMPs) from diluted cellular supernatant and showed that it significantly (~10-fold) reduced the time required to complete the assay and the sample volume used.National Institutes of Health (U.S.) (Grant GM68762)National Institutes of Health (U.S.) (Grant U54-CA112967)National Institutes of Health (U.S.) (Grant R01-EB010246)National Institutes of Health (U.S.) (Grant R01-GM081336)National Science Foundation (U.S.) (Graduate Fellowship)United States. Defense Advanced Research Projects Agency (Cipher Program