Crystallin β-b2 promotes retinal ganglion cell protection in experimental autoimmune uveoretinitis.

Crystallin βb2 (crybb2) is upregulated in regenerating retinas and in various pathological conditions of the retina, including uveoretinitis. However, the role of crybb2 in this disease is largely unknown. Therefore, we used recombinant crybb2 (rcrybb2) as intravitreal treatment of B10.

The state of the population of narrow-clawed crayfish in Lake Sevan (Armenia) under multiple stress conditions.

The narrow-clawed crayfish is a crayfish species that was accidentally introduced into the ecosystem of Lake Sevan in the 1980s. Soon it became one of the most preferred fisheries in the lake and an important link in the food chain by successfully replacing the endemic fish subspecies Sevan khramicarp (Capoeta sevangi, De Filippi, 1865) in the chain of detritivores. However, recently, the stock declines dramatically, associated with two main threats: the impact of burn-spot disease and an increase in fishing pressure on its population.

A review of focused ion beam applications in optical fibers.

Focused ion beam (FIB) technology has become a promising technique in micro- and nano-prototyping due to several advantages over its counterparts such as direct (maskless) processing, sub-10 nm feature size, and high reproducibility. Moreover, FIB machining can be effectively implemented on both conventional planar substrates and unconventional curved surfaces such as optical fibers, which are popular as an effective medium for telecommunications. Optical fibers have also been widely used as intrinsically light-coupled substrates to create a wide variety of compact fiber-optic devices by FIB milling diverse micro- and nanostructures onto the fiber surface (endfacet or outer cladding).

Alterations in Tight- and Adherens-Junction Proteins Related to Glaucoma Mimicked in the Organotypically Cultivated Mouse Retina Under Elevated Pressure.

Purpose: To scrutinize alterations in cellular interactions and cell signaling in the glaucomatous retina, mouse retinal explants were exposed to elevated pressure.

Methods: Retinal explants were prepared from C57bl6 mice and cultivated in a pressure chamber under normotensive (atmospheric pressure + 0 mm Hg), moderately elevated (30 mm Hg), and highly elevated (60 mm Hg) pressure conditions. The expression levels of proteins involved in the formation of tight junctions (zonula occludens 1 [ZO-1], occludin, and claudin-5) and adherens junctions (VE-cadherin and β-catenin) and in cell-signaling cascades (Cdc42 and activated Cdc42 kinase 1 [ACK1]), as well as the expression levels of the growth-factor receptors platelet-derived growth factor receptor beta and vascular endothelial growth factor receptors 1 and 2 (VEGFR-1, VEGFR-2) and of diverse intracellular proteins (β-III-tubulin, glial fibrillary acidic protein transcript variant 1, α-smooth muscle actin, vimentin, and von Willebrand factor VIII), were analyzed using immunohistochemistry, western blotting, and quantitative real-time polymerase chain reactions.

Elevated intraocular pressure induces neuron-specific β-III-tubulin expression in non-neuronal vascular cells.

Purpose: Pathological alterations within optic nerve axons and progressive loss of the parental retinal ganglion cell (RGC) bodies are characteristics of glaucomatous neuropathy. Abnormally elevated intraocular pressure (IOP) is thought to be the major risk factor for most forms of glaucomatous changes, while lowering of the IOP is the mainstream of treatment. However, the pathophysiological mechanisms involved in neurodegenerative changes are poorly understood.

Age-related distribution and potential role of SNCB in topographically different retinal areas of the common marmoset Callithrix jacchus, including the macula.

Evidence of an age-related increase of β-synuclein (SNCB) in several parts of the visual system including the retina has been reported. SNCB is thought to function as an antagonist of α-synuclein in neurodegenerative diseases, but the exact role of SNCB remains unclear. The presented work studies two different aspects of the onset and role of SNCB in the retinal pigment epithelium (RPE).

Transcriptomic and proteomic analysis of iris tissue and aqueous humor in juvenile idiopathic arthritis-associated uveitis.

Gene and protein expression profiles of iris biopsies, aqueous humor (AqH), and sera in patients with juvenile idiopathic arthritis-associated uveitis (JIAU) in comparison to control patients with primary open-angle glaucoma (POAG) and HLA-B27-positive acute anterior uveitis (AAU) were investigated. Via RNA Sequencing (RNA-Seq) and mass spectrometry-based protein expression analyses 136 genes and 56 proteins could be identified as being significantly differentially expressed (DE) between the JIAU and POAG group. Gene expression of different immunoglobulin (Ig) components as well as of the B cell-associated factors ID3, ID1, and EBF1 was significantly upregulated in the JIAU group as compared to POAG patients.

Retained Austenite Decomposition and Carbide Precipitation during Isothermal Tempering of a Medium-Carbon Low-Alloy Bainitic Steel.

The effect of isothermal tempering on retained austenite decomposition and carbide precipitation were investigated in a medium-carbon low-alloy bainitic steel. High-resolution dilatometry was used to perform isothermal tempering at 350 °C, 550 °C and 600 °C for different holding times up to 16 h. The decomposition of retained austenite, morphology and composition of carbides were investigated by analyzing the dilatometric curves and were confirmed through scanning and transmission electron microscopy observations.

Age-related Beta-synuclein Alters the p53/Mdm2 Pathway and Induces the Apoptosis of Brain Microvascular Endothelial Cells In Vitro.

Increased β-synuclein (Sncb) expression has been described in the aging visual system. Sncb functions as the physiological antagonist of α-synuclein (Snca), which is involved in the development of neurodegenerative diseases, such as Parkinson's and Alzheimer's diseases. However, the exact function of Sncb remains unknown.

snRPN controls the ability of neurons to regenerate axons.

Background: Retinal ganglion cells (RGCs) of mammals lose the ability to regenerate injured axons during postnatal maturation, but little is known about the underlying molecular mechanisms.

Objective: It remains of particular importance to understand the mechanisms of axonal regeneration to develop new therapeutic approaches for nerve injuries.

Methods: Retinas from newborn to adult monkeys (Callithrix jacchus)1 were obtained immediately after death and cultured in vitro.

Gradient-index optical fiber lens for efficient fiber-to-chip coupling.

A gradient-index optical fiber lens is proposed and fabricated on the tip of a single-mode fiber using focused ion beam milling. Second-order effective medium theory is used to design a gradual change in the fill factor, which ensures a parabolic effective refractive index distribution. The proposed fiber lens design is simulated via the three-dimensional finite-difference time-domain method, and demonstrated through confocal optical measurements.

[Elevated Intraocular Pressure Induces Cellular Responses in the Retinal Capillaries].

In the early diagnosis of clinical glaucoma, peripapillary bleedings were almost pathognomonic for a capillary insult. In the perfusion diagnostics, it is predominantly accepted that perfusion imbalances and IOP-induced changes occur and play a crucial role. Biomechanical peculiarities of the optic nerve head and cellular responses to astrocytes are also likely involved.

Is Angiostatin Involved in Physiological Foveal Avascularity?

Purpose: The primate central retina is characterized by an avascular fovea and well-defined perifoveal capillary plexus. Neither blood vessels nor their accompanying astrocytes enter the fovea during any stage of retinal development; a balance of angiogenic and angiostatic factors probably maintains foveal avascularity throughout life. The aim of this study was to identify potentially angiorepulsive factors involved in the development of the avascular primate retinal fovea.

The pro-inflammatory role of high-mobility group box 1 protein (HMGB-1) in photoreceptors and retinal explants exposed to elevated pressure.

To determine the role of high-mobility group box 1 protein (HMGB-1) in cellular and tissue models of elevated pressure-induced neurodegeneration, regeneration, and inflammation. Mouse retinal photoreceptor-derived cells (661W) and retinal explants were incubated either under elevated pressure or in the presence of recombinant HMGB-1 (rHMGB-1) to investigate the mechanisms of response of photoreceptors. Immunohistochemistry, western blotting, and the quantitative real-time PCR were used to examine the expression levels of immunological factors (eg, HMGB-1, receptor for advanced glycation end products (RAGE)), Toll-like receptors 2 and 4 (TLR-2, TLR-4), apoptosis-related factors (eg, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated death promoter (Bad)) as well as cytokine expression (eg, tumor necrosis factor alpha (TNF-α), interleukin (IL)-4, IL-6, and vascular endothelial growth factor (VEGF)).

Life-time expression of the proteins peroxiredoxin, beta-synuclein, PARK7/DJ-1, and stathmin in the primary visual and primary somatosensory cortices in rats.

Four distinct proteins are regulated in the aging neuroretina and may be regulated in the cerebral cortex, too: peroxiredoxin, beta-synuclein, PARK[Parkinson disease(autosomal recessive, early onset)]7/DJ-1, and Stathmin. Thus, we performed a comparative analysis of these proteins in the the primary somatosensory cortex (S1) and primary visual cortex (V1) in rats, in order to detect putative common development-, maturation- and age-related changes. The expressions of peroxiredoxin, beta-synuclein, PARK[Parkinson disease (autosomal recessive, early onset)]7/DJ-1, and Stathmin were compared in the newborn, juvenile, adult, and aged S1 and V1.

βB2-Crystallin Promotes Axonal Regeneration in the Injured Optic Nerve in Adult Rats.

The purpose of the study was to further scrutinize the potential of βB2-crystallin in supporting regeneration of injured retinal ganglion cell axons both in vitro and in vivo. Retinal explants obtained from animals after treatment either with lens injury (LI) alone or with combined LI 5 days or 3 days before or simultaneously with an optic nerve crush (ONC) were cultured for 96 h under regenerative conditions, and the regenerating axons were quantified and compared with untreated controls. These measurements were then repeated with LI replaced by intravitreal injections of γ-crystallin and β-crystallin at 5 days before ONC.

Crystallin-β-b2-overexpressing NPCs support the survival of injured retinal ganglion cells and photoreceptors in rats.

Purpose: Crystallin β-b2 (crybb2) is known to support the regeneration of retinal ganglion cell (RGC) axons in culture. We investigated whether neuronal progenitor cells (NPCs) overexpressing crybb2 (crybb2-NPC) affect secondary retinal degeneration due to optic nerve crush in vivo.

Methods: NPCS were produced by dissociation and propagation of rat embryonic neural tube and eye primordial cells at embryonic days 13.

Effects of crystallin-β-b2 on stressed RPE in vitro and in vivo.

Background: Crystallins are thought to play a cytoprotective role in conditions of cellular stress. The aim of this study was to determine the effects of crystallin-β-b2 (cryβ-b2) and crystallin-β-b3 (cryβ-b3) on ARPE-19 cells in vitro and on the retinal pigment epithelium (RPE) in vivo.

Methods: The influence of cryβ-b2 and cryβ-b3 on the viability, proliferation and dying of ARPE-19 was measured by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium assay, bromo-2-deoxyuridine assay and life/death assay.

VEGF, Ang-2 and SRIF associated abnormal postnatal development of the retinal capillary network in the Royal College of Surgeons rat.

To examine retinal angiogenesis in the Royal College of Surgeons rat (RCS) serving as a model for ischemic proliferative retinopathies at morphological, proteomic and mRNA levels in order to evaluate the interplay of morphological and molecular changes in the course of the disease. Photoreceptor degeneration was confirmed by histological cross-sections and optical coherence tomography. The capillary retinal network was visualized in RCS rats aged between 14 and 45 days (P14-P45) by perfusion with high molecular weight fluorescein isothiocyanate-labeled dextran and compared with corresponding Sprague-Dawley rats.

Optimization of transrenal DNA analysis: detection of fetal DNA in maternal urine.

Background: Fragments of DNA from cells dying throughout the body are detectable in urine (transrenal DNA, or Tr-DNA). Our goal was the optimization of Tr-DNA isolation and detection techniques, using as a model the analysis of fetal DNA in maternal urine.

Methods: We isolated urinary DNA using a traditional silica-based method and using a new technique based on adsorption of cell-free nucleic acids on Q-Sepharose resin.

Transrenal nucleic acids: from proof of principle to clinical tests.

In spite of numerous publications on potential diagnostic application of circulating DNA and transrenal nucleic acid (Tr-NA) analysis, few, if any, tests based on this technology are available in clinical labs. This delay in test development and implementation is caused, at least in part, by the deficit in robust methods for isolation of short nucleic acid fragments from bodily fluids, as well as in techniques for analyzing these fragments. We have developed a new anion exchanger-based method for the isolation of cell-free nucleic acid fragments from large volumes of bodily fluids, and analyzed these fragments by PCR techniques specially designed to amplify "ultrashort" templates.

Mycobacterium tuberculosis DNA detection in soluble fraction of urine from pulmonary tuberculosis patients.

Setting: A tertiary care and research institution in Italy.

Background: Small DNA fragments from cells dying throughout the body have been detected in urine (transrenal DNA [Tr-DNA]).

Objective: To test the hypothesis that Mycobacterium tuberculosis Tr-DNA could be detected in the urine of pulmonary tuberculosis (TB) patients.

HaCaT keratinocytes overexpressing the S100 proteins S100A8 and S100A9 show increased NADPH oxidase and NF-kappaB activities.

The calcium- and arachidonic acid (AA)-binding proteins S100A8 and S100A9 are involved in nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation in phagocytes. They are specifically expressed in myeloid cells, and are also found in epithelial cells in various (patho)physiological conditions. We have investigated the consequences of S100A8/A9 overexpression in epithelial cell lines on reactive oxygen species (ROS) generation and downstream signaling.

Novel applications of polymerase chain reaction to urinary nucleic acid analysis.

DNA fragments from cells that have died throughout the body not only appear in the bloodstream but also cross the kidney barrier into the urine. The relatively low molecular weight (150-200 bp) of this Transrenal DNA should be considered when deciding on methods of isolation and analysis. In particular, if polymerase chain reaction (PCR) is used for amplification and detection of specific sequences, then the reduction of amplicon size will significantly enhance sensitivity.

The cyclic AMP response element modulator regulates transcription of the TCR zeta-chain.

Systemic lupus erythematosus T cells display decreased amounts of TCR zeta mRNA that results in part from limited binding of the transcriptional enhancer Elf-1 to the TCR zeta promoter. We have identified a new cis-binding site for the cAMP response element (CRE) modulator (CREM) on the TCR zeta promoter, centered on the -390 nucleotide. Transfection of T cells with an antisense CREM alpha plasmid reduced the binding of CREM to the TCR zeta promoter, as shown by chromatin and reporter chromatin immunoprecipitation assays, and enhanced the production of TCR zeta mRNA and protein.