A protocol for isolation and culturing of mouse primary postmitotic photoreceptors and isolation of extracellular vesicles.

Here, we present a protocol for isolating and culturing mouse photoreceptors in a minimal, chemically defined medium free from serum. We describe steps for retina dissection, enzymatic dissociation, photoreceptor enrichment, cell culture, extracellular vesicles (EVs) enrichment, and EV ultrastructural analysis. This protocol, which has been verified for cultured cells derived from multiple murine strains, allows for the study of several aspects of photoreceptor biology, including EV isolation and nanotube formation.

ChipSeg: An Automatic Tool to Segment Bacterial and Mammalian Cells Cultured in Microfluidic Devices.

Extracting quantitative measurements from time-lapse images is necessary in external feedback control applications, where segmentation results are used to inform control algorithms. We describe ChipSeg, a computational tool that segments bacterial and mammalian cells cultured in microfluidic devices and imaged by time-lapse microscopy, which can be used also in the context of external feedback control. The method is based on thresholding and uses the same core functions for both cell types.

A Microfluidic/Microscopy-Based Platform for on-Chip Controlled Gene Expression in Mammalian Cells.

Applications of control engineering to mammalian cell biology have been recently implemented for precise regulation of gene expression. In this chapter, we report the main experimental and computational methodologies to implement automatic feedback control of gene expression in mammalian cells using a microfluidics/microscopy platform.

A tunable dual-input system for on-demand dynamic gene expression regulation.

Cellular systems have evolved numerous mechanisms to adapt to environmental stimuli, underpinned by dynamic patterns of gene expression. In addition to gene transcription regulation, modulation of protein levels, dynamics and localization are essential checkpoints governing cell functions. The introduction of inducible promoters has allowed gene expression control using orthogonal molecules, facilitating its rapid and reversible manipulation to study gene function.

IGF-1 Signaling Plays an Important Role in the Formation of Three-Dimensional Laminated Neural Retina and Other Ocular Structures From Human Embryonic Stem Cells.

We and others have previously demonstrated that retinal cells can be derived from human embryonic stem cells (hESCs) and induced pluripotent stem cells under defined culture conditions. While both cell types can give rise to retinal derivatives in the absence of inductive cues, this requires extended culture periods and gives lower overall yield. Further understanding of this innate differentiation ability, the identification of key factors that drive the differentiation process, and the development of clinically compatible culture conditions to reproducibly generate functional neural retina is an important goal for clinical cell based therapies.

Incidence and treatment of delayed symptoms of CSF leak following lumbar spinal surgery.

Purpose: Dural tear (DT) resulting in cerebrospinal fluid (CSF) leak is a common complication of spinal surgery. Most cases of DT are recognised and addressed intraoperatively; however, a small percentage of cases may present at a later stage with delayed symptoms of CSF leak, either due to an unrecognised intraoperative DT or as a result of a de novo delayed DT. Apart from few reports describing delayed symptomatic CSF leaks, most studies tend not to separate intraoperatively recognised DTs from delayed symptomatic CSF leaks.

Pain and neurological sequelae of cluster munitions on children and adolescents in South Lebanon.

This paper aims at evaluating the neurological repercussions arising from injuries sustained due to cluster munitions in children up to 18 years in South Lebanon following the 2006 conflict. Data on neurological and pain symptoms suffered during and after treatment because of sub-munitions in South Lebanon from August 2006 till late 2011 were prospectively recorded. Patients were divided into subcategories; children aged 12 and under and adolescents aged between 13 and 18.

Co-activation of the hamstrings and quadriceps during the lunge exercise.

The anterior lunge exercise is a closed chain kinetic exercise that has been developed to improve the function of the lower limb and to strengthen the hamstrings and quadriceps, simultaneously. In this study, a three-dimensional biomechanical analysis of this exercise was conducted in order to understand the mechanics of this rehabilitation activity. Experimental conditions were recorded using an active optoelectronic kinematic data capture system (OPTOTRAK), two force plates (AMTI) and electromyography (EMG).