Systematic comparison of commercial devices for temporal characterization of few-cycle laser pulses in the 500-1000 nm spectral range.

We compare multiple temporal pulse characterization techniques in three different pulse duration regimes from 15 fs to sub-5 fs, as there are no available standards yet for measuring such ultrashort pulses. To accomplish this, a versatile post-compression platform was developed, where the 100 fs near infrared pulses were post-compressed to the sub-two-cycle regime in a hybrid, three-stage configuration. After each stage, the duration of the compressed pulse was measured with the d-scan, TIPTOE and SRSI techniques and the retrieved temporal intensity profiles, spectrum and spectral phases were compared.

Accurate and comprehensive evaluation of O6-methylguanine-DNA methyltransferase promoter methylation by nanopore sequencing.

Aims: The methylation status of the O6-methylguanine-DNA methyltransferase (MGMT) promoter region is essential in evaluating the prognosis and predicting the drug response in patients with glioblastoma. In this study, we evaluated the utility of using nanopore long-read sequencing as a method for assessing methylation levels throughout the MGMT CpG-island, compared its performance to established techniques and demonstrated its clinical applicability.

Methods: We analysed 165 samples from CNS tumours, focusing on the MGMT CpG-island using nanopore sequencing.

The petawatt laser of ELI ALPS: reaching the 700 TW level at 10 Hz repetition rate.

Performance of the novel high repetition rate HF-PW laser system of ELI ALPS is presented in its first operation phase at 400 TW and 700 TW levels. Long-term operation was tested at 2.5 and 10 Hz repetition rates, where an exceptional 0.

The human lens is capable of trilineage differentiation towards osteo-, chondro-, and adipogenesis-a model for studying cataract pathogenesis.

The potential for trilineage differentiation of cells in tissues represents a model for studying disease pathogenesis and regeneration pathways. Human lens trilineage differentiation has not yet been demonstrated, and so has calcification and osteogenic differentiation of human lens epithelial cells in the whole human lens. Such changes can pose a risk for complications during cataract surgery.

Intracorneal Implantation of 3D Bioprinted Scaffolds Containing Mesenchymal Stromal Cells Using Femtosecond-Laser-Assisted Intrastromal Keratoplasty.

Injury of the cornea is a complex biological process. Regeneration of the corneal stroma can be facilitated by the presence of mesenchymal stromal cells (MSCs) and application of tissue equivalents. A new tissue-engineering strategy for corneal stroma regeneration is presented using cellularized 3D bioprinted hydrogel constructs implanted into organ cultured porcine corneas using femtosecond laser-assisted intrastromal keratoplasty.

Single thin-plate compression of multi-TW laser pulses to 3.9 fs.

Post-compression of 12-fs laser pulses with multi-TW peak power from an optical parametric chirped pulse amplification (OPCPA) system was performed by using a single thin fused silica plate in a vacuum. By optimizing the input pulses in both spatial and temporal domains, after compression with customized chirped mirrors, we achieved pulses as short as 3.87 fs, in combination with 12-mJ energy.

Subretinal Implantation of RPE on a Carrier in Minipigs: Guidelines for Preoperative Preparations, Surgical Techniques, and Postoperative Care.

Degenerative disorders of the retina (including age-related macular degeneration), which originate primarily at or within the retinal pigmented epithelial (RPE) layer, lead to a progressive disorganization of the retinal anatomy and the deterioration of visual function. The substitution of damaged RPE cells (RPEs) with in vitro cultured RPE cells using a subretinal cell carrier has shown potential for re-establishing the anatomical structure of the outer retinal layers and is, therefore, being further studied. Here, we present the principles of a surgical technique that allows for the effective subretinal transplantation of a cell carrier with cultivated RPEs into minipigs.

Subretinal Implantation of Human Primary RPE Cells Cultured on Nanofibrous Membranes in Minipigs.

Purpose: The development of primary human retinal pigmented epithelium (hRPE) for clinical transplantation purposes on biodegradable scaffolds is indispensable. We hereby report the results of the subretinal implantation of hRPE cells on nanofibrous membranes in minipigs.

Methods: The hRPEs were collected from human cadaver donor eyes and cultivated on ultrathin nanofibrous carriers prepared via the electrospinning of poly(L-lactide-co-DL-lactide) (PDLLA).

Impact of benzalkonium chloride-preserved and preservative-free latanoprost eye drops on cultured human conjunctival goblet cells upon acute exposure and differences in physicochemical properties of the eye drops.

Objective: To investigate the short-term impact on human conjunctival goblet cell (GC) survival and mucin release of acute exposure to benzalkonium chloride (BAK) preserved and preservative-free (PF) 0.005% (w/v) latanoprost (LT) eye drops, and to compare the eye drops' physicochemical properties.

Methods And Analysis: Primary GC cultures were established from human conjunctival donor tissue.

Sub-7 fs radially-polarized pulses by post-compression in thin fused silica plates.

We experimentally demonstrate the post-compression of radially polarized 25 fs pulses at 800 nm central wavelength in a multiple thin plate arrangement for the first time, to the best of our knowledge. Sub-7 fs pulses with 90 µJ energy were obtained after dispersion compensation, corresponding to a compression factor of more than 3.5.

Long-term myofibroblast persistence in the capsular bag contributes to the late spontaneous in-the-bag intraocular lens dislocation.

Late spontaneous in-the-bag intraocular lens (IOL) dislocation is a complication presenting 6 months or later after cataract surgery. We aimed to characterize the cells in the lens capsules (LCs) of 18 patients with spontaneous late in-the-bag IOL dislocation. Patients' average age was 82.

Isolation and Culture of Corneal Stromal Stem Cells.

An increasing body of evidence authenticates the benefit of corneal stroma-derived stem cells (CSSCs) in tissue engineering and regeneration oriented research, and potentially in the development of clinically relevant cellular therapies. Postmortem corneal tissue obtained from otherwise discarded material after keratoplasties is oftentimes the source of the cells for ex vivo research. Relatively easy to isolate and cultivate as well as inexpensive to culture, CSSCs now represent a well-described cell type with attributes of mesenchymal stem cells (MSCs).

Relativistic near-single-cycle optical vortex pulses from noble gas-filled multipass cells.

We propose to obtain relativistic near-single-cycle optical vortices carrying orbital angular momentum through the post-compression of Laguerre-Gaussian pulses in gas-filled multipass cells. Our simulations revealed that 30 fs optical vortex pulses centered around 800 nm with a pulse energy of millijoule level can be compressed to near-single-cycle duration with topological charges from 1 to 20 within an argon-filled cell with five passes. The spectral broadening preserves the topological charge of the input beam; the spatio-spectral couplings are also discussed.

Resveratrol as Inducer of Autophagy, Pro-Survival, and Anti-Inflammatory Stimuli in Cultured Human RPE Cells.

Unlabelled: To investigate the mechanism by which resveratrol acts upon retinal pigment epithelial (RPE) cells and to characterize its effect upon autophagy, survival, and inflammation, with consequent implications to treatment for age-related macular degeneration (AMD).

Methods: Cultured ARPE-19 cells were exposed to 10 and 50 μM resveratrol. Cell survival/death was determined by annexin-FITC/propidium iodide using flow cytometry, while autophagy was studied by detecting autophagic vacuoles formation (acridine orange and transmission electron microscopy), as well as LC3II/I ratio and p62 expression by Western blot.

Clinical and molecular markers in retinal detachment-From hyperreflective points to stem cells and inflammation.

Purpose: Retinal detachment (RD) is one of the most frequently diagnosed ophthalmologic conditions requiring prompt surgical intervention. Combination of proper surgical technique and new diagnostic markers, both clinical and molecular, can help improve the diagnosis and prognosis of RD treatment.

Methods: 12 patients with rhegmatogenous RD (rRD) were included into the study after obtaining patient consent and Regional Ethical Approval (average age: 58.

Broadband spectral characterization of the phase shift induced by population inversion in Ti:Sapphire.

The spectral phase shift of broadband amplified pulses, induced by population inversion, was measured in Ti:Sapphire at different pump fluence values. The measurement was performed for two orthogonal polarization directions and at two different crystal temperatures of 296 K and 30 K. Zero shifts and sign changes were observed in the spectral phase, which are connected to the gain spectrum of the crystal.

Effect of Isolation Technique and Location on the Phenotype of Human Corneal Stroma-Derived Cells.

Purpose: To determine the effect of the isolation technique and location upon the phenotype of human corneal stroma-derived cells (CSCs).

Methods: CSCs were isolated from the corneal stroma center and periphery using the explant or enzymatic digestion technique. The native tissue was stained for functional markers, while cultured cells were analysed by FACS.

Ex vivo 3D human corneal stroma model for Schnyder corneal dystrophy - role of autophagy in its pathogenesis and resolution.

Introduction: Multilamellar bodies (MLBs) are concentric cytoplasmic membranes which form through an autophagy-dependent mechanism. In the cornea, the presence of MLBs is associated with Schnyder corneal dystrophy (SCD). Ex vivo 3D modelling of the corneal stroma and SCD can help study pathogenesis and resolution of the disorder.

Cultivation and characterization of pterygium as an ex vivo study model for disease and therapy.

Purpose: Development of ex vivo model to study pathogenesis, inflammation and treatment modalities for pterygium.

Methods: Pterygium obtained from surgery was cultivated (3 months). Gravitational attachment method using viscoelastic facilitated adherence of graft and outgrowing cells.

Spectral phase noise analysis of a cryogenically cooled Ti:Sapphire amplifier.

The spectral phase noise of a cryogenically cooled Ti:Sapphire amplifier was analyzed by spectrally resolved interferometry. Since a relative phase difference measurement is performed, the effect of the amplifier stage can be determined with high precision. Contributions of the cooling system to the spectral phase noise were found to be below 50 mrad for both the vacuum pumps and the cryogenic system.

Liquid-cooled Ti:Sapphire thin disk amplifiers for high average power 100-TW systems.

In this work, numerical heat transfer simulations of direct water-cooled gain modules for thin disk (TD) Ti:Sapphire (Ti:Sa) power amplifiers are presented. By using the TD technique in combination with the extraction during pumping (EDP) method 100-TW class amplifiers operating around 300 W average power could be reached in the future. Single and double-sided cooling arrangements were investigated for several coolant flow velocities.

Cultivation and characterisation of the surface markers and carbohydrate profile of human corneal endothelial cells.

Background: The study aims to characterise human corneal endothelial cell (HCEnC) cultures generated by the peel-and-digest method based on their surface protein/carbohydrate expression pattern.

Methods: Quantitative polymerase chain reaction was used to compare expression of vimentin, CD90, Cytokeratin-19, ZO-1 and Claudin 14 in cultured HCEnC and cell line B4G12 versus stromal cells. Fluorescence-activated cell sorting was used to assess surface protein distribution of cultured and uncultured HCEnC.

High peak and average power Ti:sapphire thin disk amplifier with extraction during pumping.

The combination of the extraction during pumping (EDP) amplification scheme and the thin disk (TD) technology has been successfully applied to the Ti:sapphire (Ti:sa) laser medium for the first time, to the best of our knowledge. In a proof-of-principle experiment, we demonstrate high energy broadband amplification in a room temperature water cooled EDP-TD head of stretched femtosecond pulses at a 10 Hz repetition rate, instead of performing a cryogenically cooled traditional multi-pass scheme. Hence, the EDP-TD combination can overcome the limits associated with thermal effects and transverse amplified spontaneous emission, enabling Ti:sa laser systems to have a petawatt peak and hundreds of watts of average power.

Design of a thin disk amplifier with extraction during pumping for high peak and average power Ti:Sa systems (EDP-TD).

Combination of the scheme of extraction during pumping (EDP) and the Thin Disk (TD) technology is presented to overcome the limitations associated with thermal cooling of crystal and transverse amplified spontaneous emission in high average power laser systems based on Ti:Sa amplifiers. The optimized design of high repetition rate 1-10 PW Ti:Sapphire EDP-TD power amplifiers are discussed, including their thermal dynamic behavior.

Long-Term Cultures of Human Cornea Limbal Explants Form 3D Structures Ex Vivo - Implications for Tissue Engineering and Clinical Applications.

Long-term cultures of cornea limbal epithelial stem cells (LESCs) were developed and characterized for future tissue engineering and clinical applications. The limbal tissue explants were cultivated and expanded for more than 3 months in medium containing serum as the only growth supplement and without use of scaffolds. Viable 3D cell outgrowth from the explants was observed within 4 weeks of cultivation.