Multiple Critical Periods for Rapamycin Treatment to Correct Structural Defects in Tsc-1-Suppressed Brain

Tuberous sclerosis complex (TSC) is an autosomal dominant neurogenetic disorder affecting the brain and other vital organs. Neurological symptoms include epilepsy, intellectual disability, and autism. TSC is caused by a loss-of-function mutation in the TSC1 or TSC2 gene. These gene products form a protein complex and normally suppress mammalian target of rapamycin (mTOR) activity. mTOR inhibitors have been used to treat subependymal glioma (SEGA) that is a brain tumor characteristic of TSC. However, neuropathology of TSC also involves dysregulated cortical circuit formation including neuronal migration, axodendritic differentiation, and synapse formation. It is currently unknown to what extent mTOR signaling inhibitors correct an alteration in neuronal morphology that have already formed prior to the treatment. Here, we address the efficacy of rapamycin treatment on neuronal migration and dendrite formation. Using in utero electroporation, we suppressed Tsc1 expression in a fraction of neuronal progenitor cells during the fetal period. In embryonic brain slices, we found that more Tsc1-suppressed cells remained within the periventricular zone, and rapamycin treatment facilitated neuronal migration. Postnatally, Tsc1-suppressed pyramidal neurons showed more complex branching of basal dendrites and a higher spine density at postnatal day (P) 28. Aberrant arborization was normalized by rapamycin administration every other day between P1 and P13 but not P15 and P27. In contrast, abnormal spine maturation improved by rapamycin treatment between P15 and P27 but not P1 and P13. Our results indicate that there are multiple critical windows for correcting different aspects of structural abnormalities in TSC, and the responses depend on the stage of neuronal circuit formation. These data warrant a search for an additional therapeutic target to treat neurological symptoms of TSC

Particle Shape Characterisation and Classification using Automated Microscopy and Shape Descriptors in Batch Manufacture of Particulate Solids

It is known that size alone, as often defined as the volume equivalent diameter, is not sufficient for characterizing many particulate products. The shape of crystalline products can be as important as size in many applications. Traditionally particulate shape is often defined by some simple descriptors such as the maximum length and aspect ratio. Although these descriptors are intuitive, they result in loss of some information of the original shape. This paper presents a method to use principal component analysis (PCA) to derive simple latent shape descriptors from microscope images of particulate products made in batch processes, and the use of the descriptors for identification of batch to batch variations. Data from batch runs of both a laboratory crystalliser and an industrial crystallisation reactor are analysed using the approach. Qualitative and quantitative comparison with the use of traditional shape descriptors that have physical meanings and Fourier shape descriptors is also made

In-situ crystal morphology identification using imaging analysis with application to the L-glutamic acid crystallization

A synthetic image analysis strategy is proposed for in-situ crystal size measurement and shape identification for monitoring crystallization processes, based on using a real-time imaging system. The proposed method consists of image processing, feature analysis, particle sieving, crystal size measurement, and crystal shape identification. Fundamental image features of crystals are selected for efficient classification. In particular, a novel shape feature, referred to as inner distance descriptor, is introduced to quantitatively describe different crystal shapes, which is relatively independent of the crystal size and its geometric direction in an image captured for analysis. Moreover, a pixel equivalent calibration method based on subpixel edge detection and circle fitting is proposed to measure crystal sizes from the captured images. In addition, a kernel function based method is given to deal with nonlinear correlations between multiple features of crystals, facilitating computation efficiency for real-time shape identification. Case study and experimental results from the cooling crystallization of l-glutamic acid demonstrate that the proposed image analysis method can be effectively used for in-situ crystal size measurement and shape identification with good accuracy

N-cadherin: A new player in neuronal polarity

Comment on: Gärtner A, et al. EMBO J 2012; <span class="b">31</span>:1893-90

Loss of Cyclin-Dependent Kinase 5 from Parvalbumin Interneurons Leads to Hyperinhibition, Decreased Anxiety, and Memory Impairment

Perturbations in fast-spiking parvalbumin (PV) interneurons are hypothesized to be a major component of various neuropsychiatric disorders; however, the mechanisms regulating PV interneurons remain mostly unknown. Recently, cyclin-dependent kinase 5 (Cdk5) has been shown to function as a major regulator of synaptic plasticity. Here, we demonstrate that genetic ablation of Cdk5 in PV interneurons in mouse brain leads to an increase in GABAergic neurotransmission and impaired synaptic plasticity. PVCre;fCdk5 mice display a range of behavioral abnormalities, including decreased anxiety and memory impairment. Our results reveal a central role of Cdk5 expressed in PV interneurons in gating inhibitory neurotransmission and underscore the importance of such regulation during behavioral tasks. Our findings suggest that Cdk5 can be considered a promising therapeutic target in a variety of conditions attributed to inhibitory interneuronal dysfunction, such as epilepsy, anxiety disorders, and schizophrenia.National Alliance for Research on Schizophrenia and Depression (U.S.) (Young Investigator Award)National Institutes of Health (U.S.) (Grant RO1-NS051874-16)Simons Foundation (Autism Research Initiative Grant

Stereo Imaging Camera Model for 3D Shape Reconstruction of Complex Crystals and Estimation of Facet Growth Kinetics

The principle that the 3D shape of crystals that grow from a solution can be characterised in real-time using stereo imaging has been demonstrated previously. It uses the 2D images of a crystal that are obtained from two or more cameras arranged in defined angles as well as a mathematical reconstruction algorithm. Here attention is given to the development of a new and more robust 3D shape reconstruction method for complicated crystal structures. The proposed stereo imaging camera model for 3D crystal shape reconstruction firstly rotates a digitised crystal in the three-dimensional space and varies the size dimensions in all face directions. At each size and orientation, 2D projections of the crystal, according to the angles between the 2D cameras, are recorded. The contour information of the 2D images is processed to calculate Fourier descriptors and radius-based signature that are stored in a database. When the stereo imaging instrument mounted on a crystalliser captures 2D images, the images are segmented to obtain the contour information and processed to obtain Fourier descriptors and radius-based information. The calculated Fourier descriptors and radius-based signature are used to find the best matching in the database. The corresponding 3D crystal shape is thus found. Potash alum crystals that each has 26 habit faces were used as a case study. The result shows that the new approach for 3D shape reconstruction is more accurate and significantly robust than previous methods. In addition, the growth rates of {111}, {110} and {100} faces were correlated with relative supersaturation to derive models of facet growth kinetics

On-line measurement of the real size and shape of crystals in stirred tank crystalliser using non-invasive stereo vision imaging

Non-invasive stereo vision imaging technique was applied to monitoring a cooling crystallisation process in a stirred tank for real-time characterisation of the size and shape of needle-like l-glutamic acid (L-GA) β polymorphic crystals grown from solution. The instrument consists of two cameras arranged in an optimum angle that take 2D images simultaneously and are synchronised with the lighting system. Each 2D image pair is processed and analysed and then used to reconstruct the 3D shape of the crystal. The needle shaped L-GA β form crystal length thus obtained is found to be in good agreement with the result obtained from off-line analysis of crystal samples, and is about three times larger than that estimated using 2D imaging technique. The result demonstrates the advantage of 3D imaging over 2D in measurement of crystal real size and shape

Using an on-line image analysis technique to characterize sucrose crystal morphology during a crystallization run

The morphological forms and habits of crystals and agglomeration are important properties on crystallization processes. Online techniques for realtime measurement of these properties are mandatory for a better comprehension of crystal growth phenomenon. The present paper presents and describes a new online method to determine the complexity level of a crystal or a population of crystals during a crystallization process. An image analysis technique is combined with discriminant factorial analysis leading to results that allow the computation of the complexity of crystals through the parameter agglomeration degree of crystals. With this methodology, it has been possible to distinguish online and automatically among three different classes of crystals according to their complexity. It further describes the application of such methodology on the study of CaCl2, D-fructose, and D-glucose influence on the crystallization of sucrose, namely, on crystal size, morphology, and complexity. The effect of supersaturation, growth rate, and impurity concentration on the type, amount, and complexity level of the agglomerates was determined at different temperatures. The combination of image analysis and kinetic results allowed to understand better the crystallization phenomena in the presence and absence of impurities. The image analysis results suggest the possible application of this tool for process control, optimizing, by this way, laboratory and industrial crystallizers.This work was supported by Fundacao para a Ciencia e Tecnologia under program contract numbers SFRH/BD/11315/2002 and SFRH/BPD/45637/2008

How Morphological Constraints Affect Axonal Polarity in Mouse Neurons

Neuronal differentiation is under the tight control of both biochemical and physical information arising from neighboring cells and micro-environment. Here we wished to assay how external geometrical constraints applied to the cell body and/or the neurites of hippocampal neurons may modulate axonal polarization in vitro. Through the use of a panel of non-specific poly-L-lysine micropatterns, we manipulated the neuronal shape. By applying geometrical constraints on the cell body we provided evidence that centrosome location was not predictive of axonal polarization but rather follows axonal fate. When the geometrical constraints were applied to the neurites trajectories we demonstrated that axonal specification was inhibited by curved lines. Altogether these results indicated that intrinsic mechanical tensions occur during neuritic growth and that maximal tension was developed by the axon and expressed on straight trajectories. The strong inhibitory effect of curved lines on axon specification was further demonstrated by their ability to prevent formation of multiple axons normally induced by cytochalasin or taxol treatments. Finally we provided evidence that microtubules were involved in the tension-mediated axonal polarization, acting as curvature sensors during neuronal differentiation. Thus, biomechanics coupled to physical constraints might be the first level of regulation during neuronal development, primary to biochemical and guidance regulations