Zebrafish Optical Development Requires Regulated Water Permeability by Aquaporin 0.

Purpose: Optical development of the zebrafish eye relies on the movement of the highly refractive lens nucleus from an anterior to a central location in the optical axis during development. We have shown that this mechanism in turn depends on the function of Aquaporin 0a (Aqp0a), a multifunctional and extremely abundant protein in lens fiber cell membranes. Here, we probe the specific cellular functions necessary for rescuing lens nucleus centralization defects in aqp0a-/- null mutants by stable overexpression of an Aqp0 orthologue from a killifish, MIPfun.

[Syndromic growth retardation caused by impaired function of the ribosomal protein eL13].

Growth retardation for more than 2 SD below the average population or presumed familial target height is classified as a short stature and may be a clinical manifestation of a large number of disorders. The use of the latest methods of molecular genetic analysis in recent years has allowed for a better understanding of the pathogenesis of inherited forms of a short stature. One of the recently discovered mechanisms of this pathology was monoallelic mutations in RPL13 gene, leading to the development of Isidor-Toutain type spondyloepimetaphyseal dysplasia (SEDM).

KRAP is required for diffuse and punctate IP-mediated Ca liberation and determines the number of functional IPR channels within clusters.

KRas-induced actin-interacting protein (KRAP) has been identified as crucial for the appropriate localization and functioning of the inositol trisphosphate receptors (IPRs) that mediate Ca release from the endoplasmic reticulum. Here, we used siRNA knockdown of KRAP expression in HeLa and HEK293 cells to examine the roles of KRAP in the generation of IP-mediated local Ca puffs and global, cell-wide Ca signals. High resolution Ca imaging revealed that the mean amplitude of puffs was strongly reduced by KRAP knockdown, whereas the Ca flux during openings of individual IPR channels was little affected.

In vivo macromolecular crowding is differentially modulated by aquaporin 0 in zebrafish lens: Insights from a nanoenvironment sensor and spectral imaging.

Macromolecular crowding is crucial for cellular homeostasis. In vivo studies of macromolecular crowding and water dynamics are needed to understand their roles in cellular physiology and fate determination. Macromolecular crowding in the lens is essential for normal optics, and an understanding of its regulation will help prevent cataract and presbyopia.

Diabetes mellitus type 1 in children and adolescents in Moscow. Data from the Moscow Segment of the Federal Register of Diabetic Patients 2015-2020.

Background: Therapy for type 1 diabetes mellitus (T1DM) is still largely an unsolved clinical problem. Despite the introduction into clinical practice of modern insulin preparations, devices for its administration, as well as continuous monitoring of glucose levels, the goals of therapy are often not achieved. At the same time, the International Diabetes Federation (IDF) notes an increase in the prevalence and incidence of T1DM in children and adolescents in the world.

Differences in a Single Extracellular Residue Underlie Adhesive Functions of Two Zebrafish Aqp0s.

Aquaporin 0 (AQP0) is the most abundant lens membrane protein, and loss of function in human and animal models leads to cataract formation. AQP0 has several functions in the lens including water transport and adhesion. Since lens optics rely on strict tissue architecture achieved by compact cell-to-cell adhesion between lens fiber cells, understanding how AQP0 contributes to adhesion would shed light on normal lens physiology and pathophysiology.

Aquaporins Have Regional Functions in Development of Refractive Index in the Zebrafish Eye Lens.

Purpose: In the eye lens, cytosolic protein concentrations increase progressively from the periphery to the center, contributing to the gradient of refractive index (GRIN). Aquaporins are membrane proteins of lens fiber cells that regulate water transport and adhesion and interact with cytoskeletal proteins. This study investigates how these membrane proteins contribute to proper development of the lens GRIN.

Optical development in the zebrafish eye lens.

The optics of the eye is the key to a functioning visual system. The exact nature of the correlation between ocular optics and eye development is not known because of the paucity of knowledge about the growth of a key optical element, the eye lens. The sophisticated optics of the lens and its gradient of refractive index provide the superior optical quality that the eye needs and which, it is thought, has a major influence on the development of proper visual function.

Methodical inaccuracy of the Z-scan method for few-cycle terahertz pulses.

Modern sources of THz radiation generate high-intensity pulses allowing to observe nonlinear effects in this spectral range. To describe many nonlinear effects theoretically, it is necessary to know the nonlinear refractive index coefficient of optical materials. The work studies the applicability of the Z-scan method to determine the nonlinear refractive index coefficient in the THz frequency range for few-cycle pulses.

Assessment of Zebrafish Lens Nucleus Localization and Sutural Integrity.

The zebrafish is uniquely suited to genetic manipulation and in vivo imaging, making it an increasingly popular model for reverse genetic studies and for generation of transgenics for in vivo imaging. These unique capabilities make the zebrafish an ideal platform to study ocular lens development and physiology. Our recent findings that an Aquaporin-0, Aqp0a, is required for stability of the anterior lens suture, as well as for the shift of the lens nucleus to the lens center with age led us to develop tools especially suited to analyzing the properties of zebrafish lenses.

High Kerr nonlinearity of water in THz spectral range.

The values of the nonlinear refractive index coefficient for various materials in the terahertz frequency range exceed the ones in both visible and NIR ranges by several orders of magnitude. This allows to create nonlinear switches, modulators, systems requiring lower control energies in the terahertz frequency range. We report the direct measurement of the nonlinear refractive index coefficient of liquid water by using the Z-scan method with broadband pulsed THz beam.

BFSP1 C-terminal domains released by post-translational processing events can alter significantly the calcium regulation of AQP0 water permeability.

BFSP1 (beaded filament structural protein 1, filensin) is a cytoskeletal protein expressed in the eye lens. It binds AQP0 in vitro and its C-terminal sequences have been suggested to regulate the water channel activity of AQP0. A myristoylated fragment from the C-terminus of BFSP1 was found in AQP0 enriched fractions.

Aqp0a Regulates Suture Stability in the Zebrafish Lens.

Purpose: To investigate the roles of Aquaporin 0a (Aqp0a) and Aqp0b in zebrafish lens development and transparency.

Methods: CRISPR/Cas9 gene editing was used to generate loss-of-function deletions in zebrafish aqp0a and/or aqp0b. Wild type (WT), single mutant, and double mutant lenses were analyzed from embryonic to adult stages.

The modulation of the phosphorylation status of NKCC1 in organ cultured bovine lenses: Implications for the regulation of fiber cell and overall lens volume.

In previous work, we have shown the Sodium/Potassium/2 Chloride Cotransporter (NKCC1) to be a key effector of lens fiber cell volume regulation. Since others have shown that the activity of NKCC1 is regulated via its phosphorylation status, the purpose of this study was to investigate whether NKCC1 phosphorylation can be modulated in organ cultured bovine lenses, and to see how this relates to changes in lens wet weight. Western blotting was first used to confirm the expression of NKCC1, phosphorylated NKCC1 (NKCC1-P) and the regulatory kinases WNK/SPAK and phosphatases PP1/PP2A in bovine lenses at the protein level.

Calmodulin Gates Aquaporin 0 Permeability through a Positively Charged Cytoplasmic Loop.

Aquaporin 0 (AQP0), the major intrinsic protein of the eye lens, plays a vital role in maintaining lens clarity by facilitating the transport of water across lens fiber cell membranes. AQP0 reduces its osmotic water permeability constant (P) in response to increases in the external calcium concentration, an effect that is mediated by an interaction with the calcium-binding messenger protein, calmodulin (CaM), and phosphorylation of the CaM-binding site abolishes calcium sensitivity. Despite recent structural characterization of the AQP0-CaM complex, the mechanism by which CaM modulates AQP0 remains poorly understood.

Identification of the WNK-SPAK/OSR1 signaling pathway in rodent and human lenses.

Purpose: To identify whether the kinases that regulate the activity of cation chloride cotransporters (CCC) in other tissues are also expressed in rat and human lenses.

Methods: The expression of with-no-lysine kinase (WNK 1, 3, 4), oxidative stress response kinase 1 (OSR1), and Ste20-like proline alanine rich kinase (SPAK) were determined at either the transcript or protein levels in the rat and human lenses by reverse-transcriptase PCR and/or Western blotting, respectively. Selected kinases were regionally and subcellularly characterized in rat and human lenses.

[Influence of hairpin RNA, specific to the gene lawc, on expression of overlapping genes of the lawc/Trf2 complex in D. melanogaster].

We used a genetic system of the gene complex leg-arista-wing complex/TBP related factor 2 (lawc/Trf2) as a suitable model for in vivo study of operational characteristics of divergent overlapping genes. We established lines of transgenic fruit flies carrying constructs capable of expressing hairpin RNA directed at post-transcriptional suppression of the mRNA gene lawc by way of RNA-interference. The attempt to artificially suppress lawc-transcripts caused a rise in the level of expression of the gene lawc by several times.

[Recovery of genomic regions affecting lawc/Trf2 expression during Drosophila melanogaster development].

Leg-arista-wing complex (lawc) mutations affect the expression of D. melanogaster proteins homologous to a human basic transcription factor, TBP (TATA-box binding protein) Related Factor 2 (TRF2), specifically involved in development. The paper for the first time reports the application of genetic screens for various genomic regions to recover genetic interactions between the lawc/Trf2 gene and other genes and genetic loci by using Deficiency Kit lines with small deletions in total providing maximal coverage of the genome.

[Gene expression modulation is an evolutionary resource of adaptive alterations in the morphogenesis of insect limbs].

Hypomorphic mutations have been investigated of the genes spineless (ss), Distal-less (Dll), leg-arista-wing complex/TBP-related factor 2(lawc/Trf2), CG5017, and hsp 70 in order to explore the effects of their expression level on the formation of distal structures of antenna and legs of Drosophila melanogaster. We demonstrated the effect of the CG5017, hsp 70, Dll, and lawc gene transcription level on phenotypic manifestation of the ss gene mutation and the involvement of these genes into morphogenesis regulation of Drosophila melanogaster limbs. The total decrease in the level of the CG5017, hsp 70, Dll, and laws gene expression level was shown to promote a loss of the segmented pattern of distal structures of legs and antennae and a reversion of Drosophila limb morphogenesis to the evolutionarily earlier progenitors of insects.

[Clinical value of brain natriuretic peptide level in patients with atrial fibrillation].

Unlabelled: PURPOSE of this investigation was to study correlation between brain natriuretic peptide (BNP), extent of cardiac abnormalities, and six minute walk test (6-MWT) in order to asses diagnostic value of BNP in patients with trial fibrillation (F), heart failure (HF) and preserved systolic function.

Material And Methods: Sixty five patients with history of hypertension, permanent F, and shortness of breath in absence of signs of congestive hemodynamics were included into this study.

Results: Concentration of BNP in serum n the day of inclusion was significantly inversely related to results of 6-MWT, and positively related to left atrial dilation, to hospitalizations due to cardiovascular causes.

Potential in vivo UVC disinfection of catheter lumens: estimation of the doses received by the blood flow outside the catheter tip hole.

We have demonstrated that it is possible to launch UVC LED light into bacterial contaminated polymer tubes/catheters and disinfect the intraluminal space of these tubes. This can be achieved by UVC treatment of the catheters on a regular basis. Catheters are in the distal end equipped with an exit hole for administration of drugs, bloods or nutrients into the bloodstream.

[Interaction of the ss and CG5017 genes in the regulation of morphogensis of limbs in Drosophila melanogaster].

The influence of the P-element built into the area of the CG5017 gene on the mutation of the spineless (ss) gene was studied. It was shown that the insertion of the P-element decreased the level of transcription of CG5017 approximately twofold. Modulation of the level of transcription of the CG5017 gene helped demonstrate, for the first time, its influence on the phenotypic manifestation of the mutation of the ss gene, which shows their interaction in the process of regulation of morphogenesis of limbs in Drosophila melanogaster.

Expression of the sodium potassium chloride cotransporter (NKCC1) and sodium chloride cotransporter (NCC) and their effects on rat lens transparency.

Purpose: To characterize the expression patterns of the Na+-K+-Cl(-) cotransporter (NKCC) 1 and NKCC2, and the Na+-Cl(-) cotransporter (NCC) in the rat lens and to determine if they play a role in regulating lens volume and transparency.

Methods: RT-PCR was performed on RNA extracted from fiber cells to identify sodium dependent cotransporters expressed in the rat lens. Western blotting and immunohistochemistry, using NKCC1, NKCC2, and NCC antibodies, were used to verify expression at the protein level and to localize transporter expression.

[Generation of reactive oxygen species and activity of antioxidants in larva hemolymph of galleria mellonella (L.) (lepidoptera: piralidae) at development of process of encapsulation].

Activities of enzymatic antioxidants (superoxide dismutase, glutathione-S-transferase, and catalase) have been determined in hemocytes and generation of reactive oxygen species (ROS) has been studied in lymph of larvae of the wax moth Galleria mellonella at development of the process of encapsulation of nylon implants. It has been established that as soon as 15 min after piercing of cuticle with implant the capsule is formed on its surface. The active melanization of the capsule has been shown to last for 4 h.