High-throughput continuous dielectrophoretic separation of neural stem cells.

We created an integrated microfluidic cell separation system that incorporates hydrophoresis and dielectrophoresis modules to facilitate high-throughput continuous cell separation. The hydrophoresis module consists of a serpentine channel with ridges and trenches to generate a diverging fluid flow that focuses cells into two streams along the channel edges. The dielectrophoresis module is composed of a chevron-shaped electrode array. Separation in the dielectrophoresis module is driven by inherent cell electrophysiological properties and does not require cell-type-specific labels. The chevron shape of the electrode array couples with fluid flow in the channel to enable continuous sorting of cells to increase throughput. We tested the new system with mouse neural stem cells since their electrophysiological properties reflect their differentiation capacity (e.g., whether they will differentiate into astrocytes or neurons). The goal of our experiments was to enrich astrocyte-biased cells. Sorting parameters were optimized for each batch of neural stem cells to ensure effective and consistent separations. The continuous sorting design of the device significantly improved sorting throughput and reproducibility. Sorting yielded two cell fractions, and we found that astrocyte-biased cells were enriched in one fraction and depleted from the other. This is an advantage of the new continuous sorting device over traditional dielectrophoresis-based sorting platforms that target a subset of cells for enrichment but do not provide a corresponding depleted population. The new microfluidic dielectrophoresis cell separation system improves label-free cell sorting by increasing throughput and delivering enriched and depleted cell subpopulations in a single sort

Light incoherence due to quantum-gravitational fluctuations of the background space

Based on the theory of mutual coherence of light from an extended incoherent quasi-monochromatic source (providing a basis of stellar interferometry) we estimate the degree of light incoherence due to quantum-gravitational fluctuations of the background metric. It is shown that the stellar interferometry observational data considered in the literature for a last few years as a manifestation against the Planck scale quantum-gravitational fluctuations of the background metric have no chance for detecting such an effect.Comment: 5 pages; Version to appear in Astroparticle Physic

Diabetes induced by gain-of-function mutations in the Kir6.1 subunit of the KATP channel

Gain-of-function (GOF) mutations in the pore-forming (Kir6.2) and regulatory (SUR1) subunits of K(ATP) channels have been identified as the most common cause of human neonatal diabetes mellitus. The critical effect of these mutations is confirmed in mice expressing Kir6.2-GOF mutations in pancreatic β cells. A second K(ATP) channel pore-forming subunit, Kir6.1, was originally cloned from the pancreas. Although the prominence of this subunit in the vascular system is well documented, a potential role in pancreatic β cells has not been considered. Here, we show that mice expressing Kir6.1-GOF mutations (Kir6.1[G343D] or Kir6.1[G343D,Q53R]) in pancreatic β cells (under rat-insulin-promoter [Rip] control) develop glucose intolerance and diabetes caused by reduced insulin secretion. We also generated transgenic mice in which a bacterial artificial chromosome (BAC) containing Kir6.1[G343D] is incorporated such that the transgene is only expressed in tissues where Kir6.1 is normally present. Strikingly, BAC-Kir6.1[G343D] mice also show impaired glucose tolerance, as well as reduced glucose- and sulfonylurea-dependent insulin secretion. However, the response to K(+) depolarization is intact in Kir6.1-GOF mice compared with control islets. The presence of native Kir6.1 transcripts was demonstrated in both human and wild-type mouse islets using quantitative real-time PCR. Together, these results implicate the incorporation of native Kir6.1 subunits into pancreatic K(ATP) channels and a contributory role for these subunits in the control of insulin secretion

Intragenic DNA methylation: implications of this epigenetic mechanism for cancer research

Epigenetics is the study of all mechanisms that regulate gene transcription and genome stability that are maintained throughout the cell division, but do not include the DNA sequence itself. The best-studied epigenetic mechanism to date is DNA methylation, where methyl groups are added to the cytosine base within cytosine–guanine dinucleotides (CpG sites). CpGs are frequently clustered in high density (CpG islands (CGIs)) at the promoter of over half of all genes. Current knowledge of transcriptional regulation by DNA methylation centres on its role at the promoter where unmethylated CGIs are present at most actively transcribed genes, whereas hypermethylation of the promoter results in gene repression. Over the last 5 years, research has gradually incorporated a broader understanding that methylation patterns across the gene (so-called intragenic or gene body methylation) may have a role in transcriptional regulation and efficiency. Numerous genome-wide DNA methylation profiling studies now support this notion, although whether DNA methylation patterns are a cause or consequence of other regulatory mechanisms is not yet clear. This review will examine the evidence for the function of intragenic methylation in gene transcription, and discuss the significance of this in carcinogenesis and for the future use of therapies targeted against DNA methylation

Black Holes, Mergers, and the Entropy Budget of the Universe

Vast amounts of entropy are produced in black hole formation, and the amount of entropy stored in supermassive black holes at the centers of galaxies is now much greater than the entropy free in the rest of the universe. Either mergers involved in forming supermassive black holes are rare,or the holes must be very efficient at capturing nearly all the entropy generated in the process. We argue that this information can be used to constrain supermassive black hole production, and may eventually provide a check on numerical results for mergers involving black holes

Prospects of observing continuous gravitational waves from known pulsars

Several past searches for gravitational waves from a selection of known pulsars have been performed with data from the science runs of the Laser Inferometer Gravitational-wave Observatory (LIGO) gravitational wave detectors. So far these have lead to no detection, but upper limits on the gravitational wave amplitudes have been set. Here we study our intrinsic ability to detect, and estimate the gravitational wave amplitude for non-accreting pulsars. Using spin-down limits on emission as a guide we examine amplitudes that would be required to observe known pulsars with future detectors (Advanced LIGO, Advanced Virgo and the Einstein Telescope), assuming that they are triaxial stars emitting at precisely twice the known rotation frequency. Maximum allowed amplitudes depend on the stars' equation of state (e.g. a normal neutron star, a quark star, a hybrid star) and the theoretical mass quadrupoles that they can sustain. We study what range of quadrupoles, and therefore equations of state, would be consistent with being able to detect these sources. For globular cluster pulsars, with spin-downs masked by accelerations within the cluster, we examine what spin-down values gravitational wave observations would be able to set. For all pulsars we also alternatively examine what internal magnetic fields they would need to sustain observable ellipticities.Comment: version to be published in Monthly Notices of the Royal Astronomical Societ

Clinical, Biochemical and Genetic Analyses in Two Korean Patients with Medium-chain Acyl-CoA Dehydrogenase Deficiency

Medium-chain acyl-CoA dehydrogenase deficiency (MCADD) is an autosomal recessive hereditary metabolic disorder of mitochondrial fatty acid β-oxidation. It is characterized by hypoketotic hypoglycemia, hyperammonemia, seizure, coma, and sudden infant death syndrome-like illness. The most frequently isolated mutation in the acyl-CoA dehydrogenase, medium-chain (ACADM) gene of Caucasian patients with MCADD is c.985A>G, but ethnic variations exist in the frequency of this mutation. Here, we describe 2 Korean pediatric cases of MCADD, which was detected during newborn screening by tandem mass spectrometry and confirmed by molecular analysis. The levels of medium-chain acylcarnitines, including octanoylcarnitine (C8), hexanoylcarnitine (C6), and decanoylcarnitine (C10), were typically elevated. Molecular studies revealed that Patient 1 was a compound heterozygote for c.449_452delCTGA (p.Thr150ArgfsX4) and c.461T>G (p.L154W) mutations, and Patient 2 was a compound heterozygote for c.449_452delCTGA (p.Thr150ArgfsX4) and c.1189T>A (p.Y397N) mutations. We detected asymptomatic patients with MCADD by using a newborn screening test and confirmed it by ACADM mutation analysis. This report presents evidence of the biochemical and molecular features of MCADD in Korean patients and, to the best of our knowledge, this is the first report of the c.461T>G mutation in the ACADM gene

Visualization of the modeled degradation of building flooring systems in building maintenance

The development of a maintenance programme for construction projects is a highly complex and data intensive undertaking. This exercise is characterised by the lack of relevant data on the one hand and the overwhelming amount of extraneous data on the other. The uncertainties and complexities have resulted in increased conservatism in the development of lifecycle evaluation of building maintenance programing, subsequently, these programmes tend to display the symptoms of either the maintenance actions being uneconomical or fall short of providing the appropriate service to the users of the building. The current research project is based on the premise that the visual approach will facilitate a just-in-time solution to maintenance scheduling, hence, the use of virtual simulation of the building is proposed. The broader aim of this research is to develop a complete building maintenance programme through visualisation of buildings as they degrade over time. Here, the focus is on the flooring system and the manner they degrade over time. This requires a better understanding of their pattern and rate of usage. To this end, Anthroposophy and Anthropocentric descriptions of human movement pattern have been used to describe the behaviour of 'subjects' and subsequently represent the pattern and density of the degradation of flooring systems. The mathematics representing this behaviour has been developed which enables it to be embedded into the proposed overall visual building maintenance model

Stochastic Gravity: Theory and Applications

Whereas semiclassical gravity is based on the semiclassical Einstein equation with sources given by the expectation value of the stress-energy tensor of quantum fields, stochastic semiclassical gravity is based on the Einstein-Langevin equation, which has in addition sources due to the noise kernel.In the first part, we describe the fundamentals of this new theory via two approaches: the axiomatic and the functional. In the second part, we describe three applications of stochastic gravity theory. First, we consider metric perturbations in a Minkowski spacetime: we compute the two-point correlation functions for the linearized Einstein tensor and for the metric perturbations. Second, we discuss structure formation from the stochastic gravity viewpoint. Third, we discuss the backreaction of Hawking radiation in the gravitational background of a quasi-static black hole.Comment: 75 pages, no figures, submitted to Living Reviews in Relativit

Stochastic Gravity: Theory and Applications

Whereas semiclassical gravity is based on the semiclassical Einstein equation with sources given by the expectation value of the stress-energy tensor of quantum fields, stochastic semiclassical gravity is based on the Einstein-Langevin equation, which has in addition sources due to the noise kernel. In the first part, we describe the fundamentals of this new theory via two approaches: the axiomatic and the functional. In the second part, we describe three applications of stochastic gravity theory. First, we consider metric perturbations in a Minkowski spacetime, compute the two-point correlation functions of these perturbations and prove that Minkowski spacetime is a stable solution of semiclassical gravity. Second, we discuss structure formation from the stochastic gravity viewpoint. Third, we discuss the backreaction of Hawking radiation in the gravitational background of a black hole and describe the metric fluctuations near the event horizon of an evaporating black holeComment: 100 pages, no figures; an update of the 2003 review in Living Reviews in Relativity gr-qc/0307032 ; it includes new sections on the Validity of Semiclassical Gravity, the Stability of Minkowski Spacetime, and the Metric Fluctuations of an Evaporating Black Hol