The Problems of Training Youth for Future Family Life in Karakalpakstan

At the time of independence of the Republic of Uzbekistan and the Republic of Karakalpakstan there was done a great work in training young people for family life It should be noted the development of teaching materials and the introduction of the subject Psychology of Family Life into the curriculum of vocational colleges and academic lyceums which is studied in the third year when the question is actual in this age of students For educational institutions in Karakalpakstan there was designed Karakalpak version of the tutorial which is being studied by students for over 10 years It is well known that one of the leading factors in the strength of the family and a healthy psychological environment it is personal and theoretical readiness of young people to create a family There are a number of researches devoted to the study of young people s ideas about family life and the criteria of their readiness for marriage In order to strengthen the family as in a learning course Psychology of Family Life and in the life it is need effectively cultivating and use the family type of holiday the resorts sanatoriums rest homes as well as various tourist trips in the country and abroa

The Problems of Training Youth for Future Family Life in Karakalpakstan

At the time of independence of the Republic of Uzbekistan and the Republic of Karakalpakstan there was done a great work in training young people for family life It should be noted the development of teaching materials and the introduction of the subject Psychology of Family Life into the curriculum of vocational colleges and academic lyceums which is studied in the third year when the question is actual in this age of students For educational institutions in Karakalpakstan there was designed Karakalpak version of the tutorial which is being studied by students for over 10 years It is well known that one of the leading factors in the strength of the family and a healthy psychological environment it is personal and theoretical readiness of young people to create a family There are a number of researches devoted to the study of young people s ideas about family life and the criteria of their readiness for marriage In order to strengthen the family as in a learning course Psychology of Family Life and in the life it is need effectively cultivating and use the family type of holiday the resorts sanatoriums rest homes as well as various tourist trips in the country and abroa

The Unique-5 and -6 Motifs of ZO-1 Regulate Tight Junction Strand Localization and Scaffolding Properties

The proper cellular location and sealing of tight junctions is assumed to depend on scaffolding properties of ZO-1, a member of the MAGUK protein family. ZO-1 contains a conserved SH3-GUK module that is separated by a variable region (unique-5), which in other MAGUKs has proven regulatory functions. To identify motifs in ZO-1 critical for its putative scaffolding functions, we focused on the SH3-GUK module including unique-5 (U5) and unique-6 (U6), a motif immediately C-terminal of the GUK domain. In vitro binding studies reveal U5 is sufficient for occludin binding; U6 reduces the affinity of this binding. In cultured cells, U5 is required for targeting ZO-1 to tight junctions and removal of U6 results in ectopically displaced junction strands containing the modified ZO-1, occludin, and claudin on the lateral cell membrane. These results provide evidence that ZO-1 can control the location of tight junction transmembrane proteins and reveals complex protein binding and targeting signals within its SH3-U5-GUK-U6 region. We review these findings in the context of regulated scaffolding functions of other MAGUK protein

Dimerization of the Scaffolding Protein ZO-1 through the Second PDZ Domain

The tight junction protein ZO-1 is known to link the transmembrane proteins occludin, claudins, and JAMs to many cytoplasmic proteins and the actin cytoskeleton. Although specific roles for ZO-1 at the tight junction are unknown, it is widely assumed that ZO-1, together with its homologs ZO-2 and ZO-3, serves as a platform to scaffold various transmembrane and cytoplasmic tight junction proteins. Thus the manner in which the zonula occludens (ZO) proteins multimerize has implications for the protein networks they can coordinate. The purpose of our study was to determine whether ZO-1 forms homodimers and to determine the protein interaction region. Using laser light scattering and analytical centrifugation, we show that protein sequences corresponding to the NH(2)-terminal half of ZO-1 form stable homodimers with a submicromolar equilibrium dissociation constant. Analysis of the molecular weight of different truncated forms of ZO-1 revealed that the second PDZ domain is both necessary and sufficient for dimerization. This interaction does not use the beta-finger motif described for other PDZ dimers. Furthermore, ZO-1 does not dimerize via an Src homology 3 to Guk domain interaction as was demonstrated previously for MAGUKs, like PSD-95. Results from immunoprecipitation experiments with polarized Madin-Darby canine kidney epithelial cells stably transfected with full-length GFP-ZO-1 indicate that a substantial portion of ZO-1 forms homodimers in vivo. As described previously, ZO-1 also forms heterodimers with ZO-2 and ZO-3. We conclude that the dimerization of ZO proteins is unlike that of other MAGUKs and that the previously unrecognized ZO-1 homodimers may allow formation of protein networks distinct from those of heterodimers with ZO-2 and ZO-3

Nutzung von Nukleinsäure-Protein-Wechselwirkungen für die Wirkanalytik von reaktiven Species

Reaktive Species wie Superoxid, Stickstoffmonoxid oder Peroxynitrit sind in eine Vielzahl pathophysiologischer Situationen wie z.B. das Reperfusionssyndrom involviert. Besonders hohe Konzentrationen treten auf, wenn das antioxidative Abwehrsystem nicht mehr in der Lage ist, den Radikalanstieg abzufangen. Die biomedizinische Forschung hat sich auf die Mechanismen der Freisetzung dieser Species unter unterschiedlichen Streßbedingungen fokussiert. Aus diesem Grund sind verschiedene analytische Meßmethoden für den Nachweis entwickelt worden. Sensorische Methoden bieten den Vorteil einer räumlich und zeitlich aufgelösten Analyse der Einzelteilchen. Jedoch gibt es beträchtliche Wechselwirkungen zwischen den verschiedenen reaktiven Species, was die Aussagefähigkeit von Einzelmessungen in komplexen Situationen einschränkt. Hier erscheint ein Summenparameter für die Radikalwirkung im Sinne einer Wirkanalytik aussagekräftiger. Das ‘iron regulatory protein 1’ (IRP1) kann als ein solches Markerprotein betrachtet werden, dessen Konzentration Aufschluß über den zellulären Streßlevel gibt. Das Protein wird unter der Wirkung von oxidativem Streß aus dem 4Fe-4S-Enzym cytosolische Aconitase gebildet. Das entstandene IRP1 zeigt im Gegensatz zur Aconitase eine ausgeprägte Bindungsaffinität zu spezifischen m-RNA-Strukturen - den sogenannten ‘iron responsive elements’ - IREs. Dieses Verhalten kann als Grundlage für die sensorische Detektion des Markerproteins genutzt werden. In dem hier vorzustellenden experimentellen Ansatz wurde die Proteinbindung mit Hilfe der Oberflächenplasmonresonanz detektiert. Hierzu wurde eine IRE-Konsensus-Sequenz in vitro transkribiert und anschließend auf einem Carboxydextran–modifizierten Biacore-Chip immobilisiert. Eine relativ hohe Oberflächenbelegung (2000 RU /mm2) wurde sichergestellt

Post-Translational Modifications Modulate Ligand Recognition by the Third PDZ Domain of the MAGUK Protein PSD-95

The relative promiscuity of hub proteins such as postsynaptic density protein-95 (PSD-95) can be achieved by alternative splicing, allosteric regulation, and post-translational modifications, the latter of which is the most efficient method of accelerating cellular responses to environmental changes in vivo. Here, a mutational approach was used to determine the impact of phosphorylation and succinimidation post-translational modifications on the binding affinity of the postsynaptic density protein-95/discs large/zonula occludens-1 (PDZ3) domain of PSD-95. Molecular dynamics simulations revealed that the binding affinity of this domain is influenced by an interplay between salt-bridges linking the α3 helix, the β2–β3 loop and the positively charged Lys residues in its high-affinity hexapeptide ligand KKETAV. The α3 helix is an extra structural element that is not present in other PDZ domains, which links PDZ3 with the following SH3 domain in the PSD-95 protein. This regulatory mechanism was confirmed experimentally via thermodynamic and NMR chemical shift perturbation analyses, discarding intra-domain long-range effects. Taken together, the results presented here reveal the molecular basis of the regulatory role of the α3 extra-element and the effects of post-translational modifications of PDZ3 on its binding affinity, both energetically and dynamically.This research was supported by grants CVI-05915, from the Andalusian Regional Government (http://www.juntadeandalucia.es), BIO2009-13261-C02 and BIO2012-39922-C02, from the Spanish Ministry of Science and Innovation (http://www.idi.mineco.gob.es/portal/site​/MICINN/) and FEDER. JMC received a postdoctoral contract from the Spanish Ministry of Science and Innovation. CCV was a recipient of a Formación de Personal Investigador fellowship from the Spanish Ministry of Science and Innovation

ZO-1 Stabilizes the Tight Junction Solute Barrier through Coupling to the Perijunctional Cytoskeleton

ZO-1 binds numerous transmembrane and cytoplasmic proteins and is required for assembly of both adherens and tight junctions, but its role in defining barrier properties of an established tight junction is unknown. We depleted ZO-1 in MDCK cells using siRNA methods and observed specific defects in the barrier for large solutes, even though flux through the small claudin pores was unaffected. This permeability increase was accompanied by morphological alterations and reorganization of apical actin and myosin. The permeability defect, and to a lesser extent morphological changes, could be rescued by reexpression of either full-length ZO-1 or an N-terminal construct containing the PDZ, SH3, and GUK domains. ZO-2 knockdown did not replicate either the permeability or morphological phenotypes seen in the ZO-1 knockdown, suggesting that ZO-1 and -2 are not functionally redundant for these functions. Wild-type and knockdown MDCK cells had differing physiological and morphological responses to pharmacologic interventions targeting myosin activity. Use of the ROCK inhibitor Y27632 or myosin inhibitor blebbistatin increased TER in wild-type cells, whereas ZO-1 knockdown monolayers were either unaffected or changed in the opposite direction; paracellular flux and myosin localization were also differentially affected. These studies are the first direct evidence that ZO-1 limits solute permeability in established tight junctions, perhaps by forming a stabilizing link between the barrier and perijunctional actomyosin

Формирование хранилища и анализ больших данных передвижений в городе

В статье рассмотрена концепция построения хранилища данных путей передвижения для интеллектуальной транспортной системы города. Приведен математический аппарат формализованного описания транспортной системы на основе гиперсети, дуги которой являются путями передвижения. Рассмотрен рекурсивный алгоритм получения данных о путях передвижения, а также механизм использования матриц смежности по маршрутным связям. Исследования проводятся в рамках проекта АР05133699 «Исследование и разработка инновационно-телекоммуникационных технологий с использованием современных кибертехнических средств для интеллектуальной транспортной системы города». The article discusses the concept of constructing a repository of data on travel routes for the intellectual transport system of the city. The mathematical apparatus of a formalized description of a transport system based on a hypernet, whose arcs are paths of movement, is given. A recursive algorithm for obtaining data on paths of movement, as well as a mechanism for using adjacency matrices for route connections, is considered. Research is carried out as part of the project AR05133699 “Research and development of innovative telecommunication technologies using modern cyber-technical means for the intellectual transport system of the city”

Tissue plasminogen activator enhances the hypoxia/reoxygenation-induced impairment of the blood-brain barrier in a primary culture of rat brain endothelial cells.

Hemorrhagic transformation is a major complication associated with tissue plasminogen activator (tPA) therapy for ischemic stroke. We studied the effect of tPA on the blood-brain barrier (BBB) function with our in vitro monolayer model generated using rat brain microvascular endothelial cells subjected either to normoxia or to hypoxia/reoxygenation (H/R) with or without the administration of tPA. The barrier function was evaluated by the transendothelial electrical resistance (TEER), the permeability of sodium fluorescein and Evans\u27 blue-albumin (EBA), and the uptake of lucifer yellow (LY). The permeability of sodium fluorescein and EBA was used as an index of paracellular and transcellular transport, respectively. The administration of tPA increased the permeability of EBA and the uptake of LY under normoxia. It enhanced the increase in the permeability of both sodium fluorescein and EBA, the decrease in the TEER, and the disruption in the expression of ZO-1 under H/R conditions. Administration of tPA could cause an increase in the transcellular transport under normoxia, and both the transcellular and paracellular transport of the BBB under H/R conditions in vitro. Even in humans, tPA may lead to an opening of the BBB under non-ischemic conditions and have an additional effect on the ischemia-induced BBB disruption.The original publication is available at www.springerlink.co