Käytösoireisen muistisairaan lääkkeettömät hoitotyön keinot : Opas sairaanhoitajille

Aiheena opinnäytetyölle oli Käytösoireisen muistisairaan lääkkeettömät hoitotyön keinot – opas sairaanhoitajille. Opinnäytetyön tavoitteena oli tuottaa tietoa muistisairaan käytöshäiriöiden syistä ja lisäksi lisätä ymmärrystä sairaanhoitajille muistisairaan haasteellisen käytöksen lisääntymisestä. Opinnäytetyön tarkoituksena oli tuottaa yhteistyökumppanille opas, joka sisältää yleisimmät muistisairaudet ja niihin liittyvien käytöshäiriöiden ilmenemismuodot aiheuttajineen ja vaikuttavine tekijöineen. Tämän lisäksi opas sisältää hoitotyön keinoja, joiden avulla sairaanhoitaja pystyy havainnoimaan ja lieventämään muistisairaan käytösoireita. Opinnäytetyö on tehty yhteistyössä Suupohjan peruspalveluliikelaitoskuntayhtymän hoidon ja hoivan alueen kanssa. Etenevien muistisairauksien diagnostiikkaan kuuluu käytösoireiden lisääntyminen. Käytösoireita on muistisairauksien kaikissa vaiheissa ja niiden ilmaantuminen voi johtaa liialliseen ja turhaan rauhoittavien lääkkeiden määräämiseen ja käyttöön, on kuitenkin hyvä huomioida kokonaisvaltainen hoito, joka on potilaan yksilöllisen tilanteen huomioon ottava. Muistisairaiden määrä kasvaa nopeasti, varhaisen diagnosoinnin avulla pystytään ylläpitämään sairastuneen toimintakykyä ja huomioimaan sairastuneen oman elämänlaadun pysyminen hyvänä, unohtamatta hänen läheisiään. Käytösoireisen potilaan hoitolinja tulisi valita arvioimalla oireita ja selvittämällä niiden syy. Lääkkeettömän hoidon tarkoitus on, että muistisairaasta huolehditaan kokonaisvaltaisesti ja mahdollisimman hyvin hänen tarpeensa huomioon ottaen. Sairastuneen toimintakyvyn tukeminen on tärkeää, silloin hän tuntee olonsa turvatuksi ja arvostetuksi. Hyvien elämäntapojen huomioiminen, riittävän unen ja aktiviteetin turvaaminen tukevat sairastuneen tasapainon tunnetta. Käytöshäiriöiden syntyyn vaikuttaa myös ympäristössä tapahtuvat muutokset. Sairaanhoitajan on tärkeä luoda sairastuneelle tässä tilanteessa rauhallinen ja turvattu ympäristö.The subject for the thesis is non-drug nursing methods of a patient with memory disease and behavioural disorder. The aim of the thesis was to provide information on the causes of behavioural disorders of a patient with memory disease, and in addition, to increase nurses’ understanding about the negative behaviour of memory patients. The purpose of the thesis was to produce a guide containing the most common memory disorders and related manifestations of behavioural disorders, with their causes and contributing factors. In addition, the guide includes nursing tools that help the nurse to observe and mitigate the behavioural disorders of a patient with a memory disease. The thesis has been carried out in cooperation with the treatment and care area of The Suupohja Area Health and Social Services Joint Municipal Board. The diagnostics for progressive memory diseases include an increase in behavioural disorders. There are behavioural disorders at all stages of memory disorders, and their appearance may lead to the prescription and use of excessive and unnecessary medication. However, it is good to take into account the holistic treatment that is appropriate to the patient's individual condition. The number of patients with memory disease is increasing rapidly. With early diagnosis, it is pos-sible to maintain patients’ functional ability and take account of their quality of life, not forgetting their close relatives. The treatment line for the patient with behavioural disorder should be selected by evaluating the symptoms and finding out their cause. The purpose of non-drug nursing is that the patient with memory disorder is taken care of comprehensively, and his or her needs are taken into account as well as possible. Supporting the functional ability of the patient is important, and he or she feels secure and appreciated. Paying attention to a good lifestyle, ensuring adequate sleep and activity support the balance feeling of the patient. Changes in the environment also affect the appearance of behavioural disorders. It is important for the nurse to create a calm and secure environment for the patient in this situation

The association of R-Ras with FLNa enhances melanoma cell migration.

<p><b>A</b>) Transwell migration assay was used to measure the migration of M2 melanoma cells in which dominant active R-Ras G38V was expressed in stable clones expressing wild type FLNa, FLNaΔ3, or vector control with no FLNa expression. The bottom side of filters were coated with 15 µg/ml FN. Migration was expressed as the number of migrated β-Gal expressing cells. Data represent the mean ± S.D. of three independent experiments. <b>Insert</b>: Western blot of stable M2 cell clones expressing wild type FLNa, FLNaΔ3 (deletion of FLNa repeat 3), or vector control with no FLNa expression and R-Ras G38V. Tubulin was used as a loading control. Data represent at least 3 independent experiments. Statistically significant differences are reported in the graph as <i>P</i> values (Student's <i>t</i>-test), **, <i>P</i><0.01 <b>B</b>) Transwell migration assay was used to measure the migration of M2 (FLNa-) melanoma parental cell line in which FLNa, FLNaΔ3, or vector control and active R-Ras G38V and the transfection marker β-Gal were transiently transfected. Migration was expressed as the number of migrated β-Gal expressing cells. Data represent the mean ± S.D. of three independent experiments. Statistically significant differences are reported in the graph as <i>P</i> values (Student's <i>t</i>-test), **, <i>P</i><0.01; *, <i>P</i><0.04. <b>Insert</b>: Western blot of FLNa+, FLNaΔ3, or FLNa- with active R-Ras G38V expression in parental M2 (FLNa-) cell line or FLNa+, FLNaΔ3, or FLNa- cells with no active R-Ras G38V. Tubulin was used as a loading control. Data represent at least 3 independent experiments. <b>C</b>) Adhesion assay of cells expressing FLNa, FLNaΔ3, or no FLNa ± active R-Ras. Data represent at least 3 independent experiments. Top Insert: protein expression levels shown are for both A and C.</p

R-Ras and FLNa co-localize in M2 melanoma cells.

<p>FLNa and active R-Ras G38V partially co-localize in cells as examined by immunofluorescence confocal microscopy at 63X. Co-localization of FLNa and active R-Ras is displayed in yellow in the merged image. Arrows indicate co-localization. M2 (FLNa-) cells were transiently transfected with either wild type FLNa, dominant active R-Ras G38V (DA R-Ras), FLNa + DA R-Ras, FLNa + dominant negative R-Ras T43N (DN R-Ras), FLNaΔ3 (FLNa repeat 3 deleted) ± R-Ras, FLNa+R-Ras siRNA or FLNa + control scrambled siRNA and plated on FN coated glass slides. R-Ras was detected by immunostaining with an anti-R-Ras antibody followed by a Rhodamine conjugated secondary. FLNa and FLNaΔ3 were detected by immunostaining with an anti-FLNa antibody followed by Alexa Fluor 488 conjugated secondary. 63X. Data shown is representative of 3 independent experiments.</p

R-Ras binding to FLNa enhances deoxycholate-insoluble fibronectin matrix deposition.

<p><b>A</b>) Deoxycholate (DOC)-insoluble fibronectin matrix formation was examined in M2 cultures expressing active R-Ras and either wild type FLNa, FLNaΔ3 (deletion of FLNa repeat 3), or M2 (FLNa-) control cell cultures. FLNa cultures with knockdown of endogenous R-Ras using R-Ras siRNA were assessed. Control scrambled siRNA and cultures not expressing FLNa (FLNa-) were also assessed. 48 hr after the addition of human FN to the wells, the DOC-insoluble material was subjected to SDS-PAGE analysis under reducing conditions and FN was detected by immunostaining. FN = Fibronectin. Vimentin was used as a loading control. <b>B</b>) Fold induction of matrix was normalized to M2 (FLNa-) control cultures. Graph shown represents pooled data from three independent experiments. Error bars represent S.E.M. Statistically significant differences are reported in the graph as <i>P</i> values (Student's <i>t</i>-test), **, <i>P</i><0.05. <b>C</b>) Knockdown of endogenous R-Ras using siRNA or scrambled control siRNA is demonstrated via Western blot. 60 hr post transfection cells were lysed, run on SDS-PAGE gel and immunoblotted for R-Ras using a polyclonal anti-R-Ras antibody. Tubulin was used as a loading control. Results are representative of 3 independent experiments.</p

R-Ras associates with FLNa at repeat 3.

<p><b>A</b>) Yeast two hybrid screen of R-Ras binding to FLNa. Colonies from + histidine (+his) plates obtained by co-transformation of the L40 yeast strain with BTM116R-Ras-CAAX^- or negative control BTM116-lamin C and VP16-FLNa (nucleotides 1699–1995). The plate was photographed after 3 days of growth. <b>B</b>) R-Ras binds FLNa. GST fusion proteins derived from indicated FLNa fragments were incubated with M2 (FLNa-) cell total lysates. Bound R-Ras was detected by Western blotting for R-Ras. Results are representative of 3 independent experiments. Coomassie stained gel containing purified FLNa fragments is shown to confirm FLNa fragment protein expression. <b>C</b>) R-Ras and FLNa co-immunoprecipitate in M2 melanoma cells. M2 cells were co-transfected with expression vectors for active R-Ras G38V, His-tagged wild type FLNa, His-tagged FLNaΔ3 (deletion of FLNa repeat 3) or vector alone. 48 h post transfection cells were lysed, immunoprecipitated (IP) with anti-His conjugated IgG beads or IgG beads alone and immunoprecipitates were analyzed by immunoblotting with an anti-R-Ras antibody. Input R-Ras and FLNa levels are shown (Bottom). Results are representative of 3 independent experiments. <b>D</b>) The domain structure of FLNa and the location of R-Ras binding site as determined by yeast two hybrid screen. The β-integrin subunits, the actin-binding domain, and the dimerization domain at the C-terminus have been established previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011269#pone.0011269-Pfaff1" target="_blank">[5]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011269#pone.0011269-Gorlin1" target="_blank">[49]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011269#pone.0011269-Loo1" target="_blank">[50]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0011269#pone.0011269-Sharma1" target="_blank">[51]</a>. FLNa fragments repeats 1–10, 11–16, and 17–23 + dimerization domain are shown on the left.</p

The association between R-Ras and FLNa enhances integrin activation.

<p><b>A</b>) M2 cells express a subset of integrins on their surface. FACS analysis of a subset of integrins expressed on the surface of M2 cells. M2 cells express α5 and β1 subunits on their surface but not αvβ3. <b>B</b>) Forced expression of activated R-Ras promotes integrin activation in M2 cells as determined by the 3Fn-(9–11) activation assay. M2 (FLNa-) cells were transiently transfected with expression vectors encoding GFP as transfection reporter and FLNa, constitutively active R-Ras G38V alone or ±FLNa or FLNaΔ3. Cells were harvested and analyzed by two-color FACS for transfected cells and 3FN-(9–11) binding. Shown on the <i>Y-Axis</i> is mean activation index ±S.E. of three independent experiments. Constitutively expressed active R-Ras G38V and FLNa increased the activation index more than R-Ras alone. FLNaΔ3 did not block R-Ras-mediated activation compared with controls. Statistically significant differences are reported in the graph as <i>P</i> values (Student's <i>t</i>-test), *, <i>P</i><0.05 **, <i>P</i><0.04 <b>C</b>) Western blot of R-Ras levels when transiently transfected with R-Ras G38V. Tubulin was used as a loading control. Data represent at least 3 independent experiments.</p

R-Ras binding to FLNa enhances fibronectin matrix assembly.

<p>Fibronectin matrix assembly in cultures expressing R-Ras G38V and either wild type FLNa (<b>A</b>), FLNaΔ3 (deletion of FLNa repeat 3; <b>B</b>), or cell cultures that do not express FLNa (FLNa-; <b>C</b>) were assessed. FLNa cultures expressing PcDNA3 vector and control scrambled siRNA (<b>D</b>) or knockdown of endogenous R-Ras using R-Ras siRNA (<b>E</b>) or control cultures not expressing FLNa (<b>F</b>) were also assessed. 24 hr after plating the culture media was replaced with media containing human FN (100 µg/ml) and cells were cultured for a further 48 hr before fibronectin matrix was assessed by immunostaining for FN. Each experiment was repeated a minimum of 3X.</p

Additional file 2: Figure S1. of Tartrate-resistant acid phosphatase (TRAP/ACP5) promotes metastasis-related properties via TGFβ2/TβR and CD44 in MDA-MB-231 breast cancer cells

Distribution of unique peptides across fractions generated by HiRIEF separation on IPG 2.5–3.7 strips. Number of unique peptides and phospho-peptides identified across fractions in each biological replicate (A) and (B), displayed by number of phosphorylations. Fraction numbering proceeds from the acidic end towards the basic end of the strips. Multiply phosphorylated peptides are identified prevalently in the first 30 fractions in the IPG 2.5–3.7 strip. (PDF 148 kb

Additional file 1: Material and Methods. of Tartrate-resistant acid phosphatase (TRAP/ACP5) promotes metastasis-related properties via TGFβ2/TβR and CD44 in MDA-MB-231 breast cancer cells

Additional explanations on Material and Methods including descriptions of gene expression analysis, cell lysis, SILAC labeling, protein extraction for mass spectrometric analyses, protein digestion, Tandem Mass Tag labeling, liquid chromatography tandem mass spectrometric analyses and proteomics database search, protein and phospho-peptide ratios calculation. (DOCX 32 kb

Additional file 4: Table S1. of Tartrate-resistant acid phosphatase (TRAP/ACP5) promotes metastasis-related properties via TGFβ2/TβR and CD44 in MDA-MB-231 breast cancer cells

Phosphorylation sites significantly regulated in TRAP3high cells compared to control (ctrl) cells. Unique phosphorylation sites are defined by a sequence window of 15 amino acids centered at the phosphorylated residue. The column “Protein class” describes to which of the examined protein class a protein belongs to. Examined proteins classes include protein kinases, protein phosphatases, ubiquitin and UBL system enzymes and transcription factors. The column “Functional site” describes whether the phospho-site has a known regulatory function based on the information available on the database PhosphoSitePlus. (XLSX 48 kb