Особливості зображення образу коханої в інтимній ліриці Григорія Чупринки (Features of the image of cohana in immediate lyrics Grigoria Chuprynki)

У статті аналізується поезія Григорія Чупринки, де було розглянуто образ коханої жінки ліричного героя. Відтворено портрет героїні; розкрито символіку її асоціативних образів, основні риси характеру. Визначено для поетичної практики автора засоби творення художнього образу, а також його стильові особливості в любовній ліриці. (The article analyzes the poetry of Grigoria Chuprinky, where the image of the beloved woman of the lyrical hero was considered. With insuffi cient study of the love lyrics of Grigoria Chuprinky, it can be argued that the theme of love occupied an important place in the work of the artist. In the poetry, the author does not depict the physical image of the woman himself, but her presence is quite tangible, and thanks to the caressful words, we feel the sincerity and tenderness of the feelings of a lover. Poetry is permeated with the motive of cleansing from sin as a «sacred love» of a beloved, and love acts as the only feeling that is able to atone for all the sins of the earth. In order to reveal the image of a beloved author as much as possible, and to the musicality of poetry, through the combination of assonances and alliterations, he forms the elegiac mood of the poem. In the manner of his beloved, Chuprinka combined the best features: she is beautiful, her character is changing, and in addition she hopes to realize a cherished dream. However, the image of a woman is quite contrasted and ambiguous: she is able to heal the mental wounds of her husband, while, at the same time, she may mortally strike his eyes with her eyes. The author is using the contrast: creates a portrait of a woman from the inside, based on the psychological depth and mystery of herinner world. A portrait of a heroine is restored; the symbols of its associative images, the main features of the character are revealed. The poetic practice of the author of the means of creating an artistic image, as well as his stylistic features in love poetry, is determined.

Ubiquitination increases parkin activity to promote autophagic α-synuclein clearance.

Parkinson's disease (PD) is a movement disorder associated with genetic and age related causes. Although autosomal recessive early onset PD linked to parkin mutations does not exhibit α-Synuclein accumulation, while autosomal dominant and sporadic PD manifest with α-Synuclein inclusions, loss of dopaminergic substantia nigra neurons is a common denominator in PD. Here we show that decreased parkin ubiquitination and loss of parkin stability impair interaction with Beclin-1 and alter α-Synuclein degradation, leading to death of dopaminergic neurons. Tyrosine kinase inhibition increases parkin ubiquitination and interaction with Beclin-1, promoting autophagic α-Synuclein clearance and nigral neuron survival. However, loss of parkin via deletion increases α-Synuclein in the blood compared to the brain, suggesting that functional parkin prevents α-Synuclein release into the blood. These studies demonstrate that parkin ubiquitination affects its protein stability and E3 ligase activity, possibly leading to α-Synuclein sequestration and subsequent clearance

Novel quantitative trait Loci controlling development of experimental autoimmune encephalomyelitis and proportion of lymphocyte subpopulations.

The B10.RIII mouse strain (H-2r) develops chronic experimental autoimmune encephalomyelitis (EAE) upon immunization with the myelin basic protein 89–101 peptide. EAE was induced and studied in a backcross between B10.RIII and the EAE-resistant RIIIS/J strain (H-2r), and a complete genome scan with microsatellite markers was performed. Five loci were significantly linked to different traits and clinical subtypes of EAE on chromosomes 1, 5, 11, 15, and 16, three of the loci having sex specificity. The quantitative trait locus on chromosome 15 partly overlapped with the Eae2 locus, previously identified in crosses between the B10.RIII and RIIIS/J mouse strains. The loci on chromosomes 11 and 16 overlapped with Eae loci identified in other mouse crosses. By analyzing the backcross animals for lymphocyte phenotypes, the proportion of B and T cells in addition to the levels of CD4+CD8- and CD4-CD8+ T cells and the CD4+/CD8+ ratio in spleen were linked to different loci on chromosomes 1, 2, 3, 5, 6, 11, and 15. On chromosome 16, we found significant linkage to spleen cell proliferation. Several linkages overlapped with the quantitative trait loci for disease phenotypes. The identification of subphenotypes that are linked to the same loci as disease traits could be most useful in the search for candidate genes and biological pathways involved in the pathological process

Fractalkine Mediates Communication between Pathogenic Proteins and Microglia: Implications of Anti-Inflammatory Treatments in Different Stages of Neurodegenerative Diseases

The role of inflammation in neurodegenerative diseases has been widely demonstrated. Intraneuronal protein accumulation may regulate microglial activity via the fractalkine (CX3CL1) signaling pathway that provides a mechanism through which neurons communicate with microglia. CX3CL1 levels fluctuate in different stages of neurodegenerative diseases and in various animal models, warranting further investigation of the mechanisms underlying microglial response to pathogenic proteins, including Tau, β-amyloid (Aβ), and α-synuclein. The temporal relationship between microglial activity and localization of pathogenic proteins (intra- versus extracellular) likely determines whether neuroinflammation mitigates or exacerbates disease progression. Evidence in transgenic models suggests a beneficial effect of microglial activity on clearance of proteins like Aβ and a detrimental effect on Tau modification, but the role of CX3CL1 signaling in α-synucleinopathies is less clear. Here we review the nature of fractalkine-mediated neuronmicroglia interaction, which has significant implications for the efficacy of anti-inflammatory treatments during different stages of neurodegenerative pathology. Specifically, it is likely that anti-inflammatory treatment in early stages of disease during intraneuronal accumulation of proteins could be beneficial, while anti-inflammatory treatment in later stages when proteins are secreted to the extracellular space could exacerbate disease progression

TKI increases parkin ubiquitination and interaction with Beclin-1.

<p><i>In Situ</i> PLA in 20 µm thick mouse brain sections shows <b>A</b>) parkin-Beclin-1 interaction in the striatum of C57BL/6 control, <b>B</b>) parkin^−/− mice, <b>C</b>) transgenic A53T mice treated with daily I.P injection for 3 weeks of DMSO <b>D</b>) 10 mg/kg nilotinib and <b>E</b>) 5 mg/kg bosutinib (n = 5). <i>In situ</i> PLA in 20 µm thick mouse brain sections showing parkin ubiquitination via <b>F</b>) parkin-ubiquitin interaction in the striatum of C57BL/6 control, <b>G</b>) transgenic A53T mice treated with daily I.P injection for 3 weeks of DMSO <b>H</b>) 10 mg/kg nilotinib and <b>I</b>) 5 mg/kg bosutinib (n = 5). <b>J</b>) Histograms represent parkin ubiquitination in B35 rat neuroblastoma cells treated with either 10 µM nilotinib or 1 µM bosutinib and 20 µM proteasome inhibitor MG132. Parkin ubiquitination was observed with WT ubiquitin not the K0 control (n = 6), compared to recombinant E1-E2-E3 control. Asterisk indicates significantly different, means±SEM, ANOVA, Newman Keuls, n = 6) and <b>K</b>). immnunoprecipitation of cell extracts with anti-ubiquitin antibodies and WB with parkin showing ubiquitinated proteins (n = 5).</p

Parkin co-localizes with Beclin-1 in human brain but their interaction is decreased in sporadic PD.

<p>Immunostaining of paraffin embedded 20 µm thick human midbrain sections <b>A</b>) shows parkin expression in DAPI stained neurons co-labeled with GFAP. Inset is higher magnification showing parkin expression. <b>B</b>) TH⁺ neurons counterstained with DAB in serial midbrain sections of healthy human subjects. <b>C</b>) <i>In situ</i> proximity ligation assay (PLA) shows human parkin-Beclin-1 interaction in control midbrain neurons (n = 7). <b>D</b>) Parkin expression in DAPI stained neurons co-labeled with GFAP indicating active astrocytes. Inset is higher magnification showing cytosolic parkin accumulation. <b>E</b>) TH⁺ neurons counterstained with DAB in serial midbrain sections of post-mortem PD brains. <b>F)</b><i>In situ</i> PLA shows reduced human parkin-Beclin-1 interaction in midbrain neurons in PD brains (n = 9). <b>G</b>). parkin and <b>H</b>) and Beclin-1 labeling are co-localized <b>I</b>) in human midbrain sections (n = 7). <b>J</b>) shows alteration of parkin expression and <b>K</b>) Beclin-1 staining with <b>L</b>) reduced co-localization in sporadic PD brains (n = 9). <b>M</b>) graph represents stereological counting of C and F <i>in situ</i> PLA and <b>N</b>) WB analysis on 4−12% NuPage SDS gel showing beclin-1 levels relative to actin and immunoprecipitation in post-mortem human brain tissues. Asterisk indicates significantly different, means±SEM, ANOVA, Newman Keuls.</p

TKIs stimulate autophagic α-Synuclein clearance in A53T mice.

<p>ELISA measurement of human α-Synuclein in the <b>A</b>) brain and <b>B</b>) blood of 1 month old A53T mice injected I.P. with either 5 mg/kg or 1 mg/kg bosutinib once every other day for 6 weeks (n = 10). <b>C</b>). Insert shows separation of AVs and lysosomal fractions. Histograms represent subcellular fractionation of total brain lysates from A53T mice and ELISA of human α-Synuclein in AVs in 1 and 5 months old A53T brains after daily I.P. injection with 10 mg/kg nilotinib or 5 mg/kg bosutinib for 3 weeks (n = 5). <b>D</b>). ELISA showing parkin level in total brain extracts of A53T mice compared to WT and parkin^−/− in the presence of nilotinib and bosutinib (n = 10). <b>E</b>). Histograms represent subcellular fractionation of total brain lysates from A53T mice and parkin ELISA in AVs in 1 and 5 months old A53T brains after daily I.P. injection with 10 mg/kg nilotinib or 5 mg/kg bosutinib for 3 weeks (n = 5). Asterisk indicates significantly different, means±SEM, ANOVA, Newman Keuls. F) WB analysis on 4−12% NuPage SDS gel showing parkin and α-Synuclein levels relative to actin in A53T mice treated with either DMSO or bosutinib.</p

TKI clears α-Synuclein and protects TH⁺ neurons in a parkin-dependent manner.

<p>Immunostaining of 20 µm thick mouse brain sections showing human α-Synuclein counterstained with DAB in the SN of lentiviral injected <b>A</b>) LacZ WT mice treated with DMSO for 3 weeks, <b>B</b>) LacZ WT treated with 5 mg/kg bosutinib, <b>C</b>) human WT α-Synuclein in WT mice treated with DMSO, and <b>D</b>) human WT α-Synuclein in WT mice treated with 5 mg/kg bosutinib (n = 10). TH⁺ labeling counterstained with DAB in the SN of lentiviral injected <b>E</b>) LacZ WT mice treated with DMSO for 3 weeks, <b>F</b>) LacZ WT treated with 5 mg/kg bosutinib, <b>G</b>) human WT α-Synuclein in WT mice treated with DMSO, and <b>H</b>) human WT α-Synuclein in WT mice treated with 5 mg/kg bosutinib (n = 10). Human α-Synuclein in the SN of parkin^−/− mice counterstained with DAB injected with <b>I)</b> LacZ treated with DMSO, or human α-Synuclein treated with <b>J</b>) DMSO, <b>K</b>) 5 mg/kg bosutinib and <b>L</b>) 10 mg/kg nilotinib for 3 weeks (n = 10). Staining of TH⁺ neurons in the SN of parkin^−/− mice counterstained with DAB injected with <b>M</b>) LacZ treated with DMSO <b>O</b>) is higher magnification, or lentiviral α-Synuclein treated with <b>N</b>) DMSO, <b>R</b>) is a higher magnification <b>O</b>) 5 mg/kg bosutinib, <b>S</b>) is a higher magnification and <b>P</b>) 10 mg/kg nilotinib, <b>T</b>) is a higher magnification (n = 10). <b>U</b>) graph represents stereological counting and <b>V</b>) RT-PCR showing equal amount of α-Synuclein mRNA relative to GADPH in both parkin^−/− and WT mice with and without TKIs (n = 10). Asterisk indicates significantly different, means±SEM, ANOVA, Newman Keuls.</p