Consistent with recent reports indicating that neurons differentiated in vitro from human induced pluripotent stem cells (hiPSCs) are immature relative to those in the human brain (1, 2), gene expression comparisons of our hiPSC-derived neurons to the Allen BrainSpan Atlas indicate that they most resemble fetal brain tissue. This finding suggests that, rather than modeling the late features of schizophrenia (SZ), hiPSC-based models may be better suited for the study of disease predisposition. We now report that a significant fraction of the gene signature of SZ hiPSC-derived neurons is conserved in SZ hiPSC neural progenitor cells (NPCs). We used two, independent discovery-based approaches - microarray gene expression and stable isotope labeling by amino acids in cell culture (SILAC) quantitative proteomic mass spectrometry analyses – to identify cellular phenotypes in SZ hiPSC NPCs from four SZ patients. From our findings that SZ hiPSC NPCs show abnormal gene expression and protein levels related to cytoskeletal remodeling and oxidative stress, we predicted, and subsequently observed, aberrant migration and increased oxidative stress in SZ hiPSC NPCs. This approach, consisting of reproducible phenotypes identified through scalable assays, can be applied to expanded cohorts of SZ patients, making it a potentially valuable tool with which to study the developmental mechanisms contributing to SZ.

Prenatal exposure of the developing brain to various environmental challenges increases susceptibility to late-onset of neuropsychiatric dysfunction; still the underlying mechanisms remain obscure. Here we show that exposure of embryos to a variety of environmental factors such as alcohol, methylmercury and maternal seizure activates HSF1 in cerebral cortical cells. Furthermore, Hsf1 deficiency in the mouse cortex exposed in utero to subthreshold levels of these challenges causes structural abnormalities and increases seizure susceptibility after birth. In addition, we found that human neural progenitor cells differentiated from induced pluripotent stem cells derived from schizophrenia patients show higher variability in the levels of HSF1 activation induced by environmental challenges compared to controls. We propose that HSF1 plays a crucial role in the response of brain cells to prenatal environmental insults and may be a key component in the pathogenesis of late–onset neuropsychiatric disorders.

We report the detection of slice-selective electron spin resonance with an external magnetic field gradient comparable to local interatomic gradients, using the techniques of magnetic resonance force microscopy. An applied microwave field modulated the spin-gradient force between a paramagnetic DPPH sample and a micrometer-scale ferromagnetic tip on a force microscope cantilever. A sensitivity equivalent to 184 polarized electron moments in a one-Hertz detection bandwidth was attained. We mapped the tip magnetic field with a resonant slice thickness of order one nanometer, thereby demonstrating magnetic resonance on length scales comparable to molecular dimensions.

Thermal fluctuations generate magnetic noise in the vicinity of any conductive and/or magnetically permeable solid. This magnetic noise plays a fundamental role in the design of spintronic devices: namely, it sets the time scale during which electron spins retain their coherence. This paper presents a rigorous classical and quantum analysis of thermal magnetic noise, together with prac- tical engineering examples. Starting with the fluctuation-dissipation theorem and Maxwell's equations, a closed-form expression for the spectral density of thermal magnetic noise is derived. Quantum decoherence, as induced by thermal magnetic noise, is analyzed via the independent oscillator heat bath model of Ford et al. The resulting quantum Langevin equations yield closed-form expressions for the spin relaxation times. For realistic experiments in spin-tronics, magnetic resonance force microscopy, Bose-Einstein condensates, atomic physics, and solid-state quantum computing, the predicted relaxation rates are rapid enough that substantial experimental care must be taken to minimize them. At zero temperature, the quantum entanglement between a spin state and a thermal reservoir is computed. The same Hamiltonian matrix elements that govern fluctuation and dissipation are shown to also govern entanglement and renormalization, and a specific example of a fluctuation-dissipation-entanglement theorem is constructed. We postulate that this theorem is independent of the detailed structure of thermal reservoirs, and therefore expresses a general thermodynamic principle.

Magnetic resonance force microscopy (MRFM) images the three-dimensional spatial distribution of resonant spins by mechanical force detection. Image reconstruction in MRFM is challenging because the resonance occurs in a strongly curved shell that extends beyond the scan range. In contrast with conventional magnetic resonance imaging, where Fourier techniques work well, the curved-shell resonant geometry inherent to MRFM requires novel reconstruction methods. Here, we show the application of iterative reconstruction in an electron spin resonance imaging experiment with 80 nm voxels. The reconstructed image has a total scan volume of 0.5 cubic micrometers, and was generated by a magnetic resonant shell with a curvature radius of 2.3 μm. The imaged object was a paramagnetically doped solid with an obliquely tilted surface. The reconstructed image correctly identified the location and orientation of the surface, and mapped the spin distribution within the solid. Applications of MRFM include three-dimensional nanometer-scale mapping of dopant distributions in semiconductors, studies of magnetism of thin films, and spin diffusion physics. An ultimate goal of MRFM is the direct observation of molecular structure at the atomic scale.

Poly(dimethylsiloxane) (PDMS) is a common material used in fabricating microfluidic devices. The predominant PDMS fabrication method, soft lithography, relies on photolithography for fabrication of micropatterned molds. In this technical note, we report an alternative molding technique using microscale PLasma Activated Templating (microPLAT). The use of photoresist in soft lithography is replaced by patterned water droplets created using microPLAT. When liquid PDMS encapsulates patterned water and then solidifies, the cavities occupied by water become structures such as microchannels. Using this method, device fabrication is less time consuming, more cost efficient and flexible, and ideal for rapid prototyping. An additional important feature of the water-molding process is that it yields structural profiles that are difficult to achieve using photolithography.

The pharmacokinetics and pharmacodynamics of ziconotide were assessed over a 48-hour period following intrathecal (i.t.) administration (1, 5, 7.5, or 10 micrograms) to 22 patients with chronic, nonmalignant pain. Plasma and cerebrospinal fluid (CSF) samples were obtained over a 24-hour period. Analgesic efficacy was monitored using Visual Analog Scale of Pain Intensity (VASPI) and Category Pain Relief Scores (CPRS) measurements. Pharmacokinetic (PK) parameters were calculated by noncompartmental methods. Plasma ziconotide data were insufficient for PK calculations. In CSF, the median half-life of ziconotide was 4.5 hours. The median CSF clearance and volume of distribution were 0.26 mL/min and 99 mL, respectively. CSF pharmacokinetics of ziconotide were linear, based on cumulative exposure and peak CSF concentrations. A dose-related analgesia was observed. Pharmacokinetic-pharmacodynamic efficacy and safety analyses showed that higher CSF ziconotide concentrations were generally associated with analgesia and increased incidence of nervous system adverse events following a 1-hour i.t. infusion.

This work describes an enhancement to the loop-mediated isothermal amplification (LAMP) reaction which results in improved performance. Enhancement is achieved by adding a new set of primers to conventional LAMP reactions. These primers are termed "swarm primers" based on their relatively high concentration and their ability to create new amplicons despite the theoretical lack of single-stranded annealing sites. The primers target a region upstream of the FIP/BIP primer recognition sequences on opposite strands, substantially overlapping F1/B1 sites. Thus, despite the addition of a new primer set to an already complex assay, no significant increase in assay complexity is incurred. Swarm priming is presented for three DNA templates: Lambda phage, Synechocystis sp. PCC 6803 rbcL gene, and human HFE. The results of adding swarm primers to conventional LAMP reactions include increased amplification speed, increased indicator contrast, and increased reaction products. For at least one template, minor improvements in assay repeatability are also shown. In addition, swarm priming is shown to be effective at increasing the reaction speed for RNA amplification via RT-LAMP. Collectively, these results suggest that the addition of swarm primers will likely benefit most if not all existing LAMP assays based on state-of-the-art, six-primer reactions.