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

R-Ras Regulates Migration through an Interaction with Filamin A in Melanoma Cells

Changes in cell adhesion and migration in the tumor microenvironment are key in the initiation and progression of metastasis. R-Ras is one of several small GTPases that regulate cell adhesion and migration on the extracellular matrix, however the mechanism has not been completely elucidated. Using a yeast two-hybrid approach we sought to identify novel R-Ras binding proteins that might mediate its effects on integrins.We identified Filamin A (FLNa) as a candidate interacting protein. FLNa is an actin-binding scaffold protein that also binds to integrin β1, β2 and β7 tails and is associated with diverse cell processes including cell migration. Indeed, M2 melanoma cells require FLNa for motility. We further show that R-Ras and FLNa interact in co-immunoprecipitations and pull-down assays. Deletion of FLNa repeat 3 (FLNaΔ3) abrogated this interaction. In M2 melanoma cells active R-Ras co-localized with FLNa but did not co-localize with FLNa lacking repeat 3. Thus, activated R-Ras binds repeat 3 of FLNa. The functional consequence of this interaction was that active R-Ras and FLNa coordinately increased cell migration. In contrast, co-expression of R-Ras and FLNaΔ3 had a significantly reduced effect on migration. While there was enhancement of integrin activation and fibronectin matrix assembly, cell adhesion was not altered. Finally, siRNA knockdown of endogenous R-Ras impaired FLNa-dependent fibronectin matrix assembly.These data support a model in which R-Ras functionally associates with FLNa and thereby regulates integrin-dependent migration. Thus in melanoma cells R-Ras and FLNa may cooperatively promote metastasis by enhancing cell migration

Bone Alkaline Phosphatase and Tartrate-Resistant Acid Phosphatase: Potential Co-regulators of Bone Mineralization

Phosphorylated osteopontin (OPN) inhibits hydroxyapatite crystal formation and growth, and bone alkaline phosphatase (BALP) promotes extracellular mineralization via the release of inorganic phosphate from the mineralization inhibitor inorganic pyrophosphate (PPi). Tartrate-resistant acid phosphatase (TRAP), produced by osteoclasts, osteoblasts, and osteocytes, exhibits potent phosphatase activity towards OPN; however, its potential capacity as a regulator of mineralization has not previously been addressed. We compared the efficiency of BALP and TRAP towards the endogenous substrates for BALP, i.e., PPi and pyridoxal 5-phosphate (PLP), and their impact on mineralization in vitro via dephosphorylation of bovine milk OPN. TRAP showed higher phosphatase activity towards phosphorylated OPN and PPi compared to BALP, whereas the activity of TRAP and BALP towards PLP was comparable. Bovine milk OPN could be completely dephosphorylated by TRAP, liberating all its 28 phosphates, whereas BALP dephosphorylated at most 10 phosphates. OPN, dephosphorylated by either BALP or TRAP, showed a partially or completely attenuated phosphorylation-dependent inhibitory capacity, respectively, compared to native OPN on the formation of mineralized nodules. Thus, there are phosphorylations in OPN important for inhibition of mineralization that are removed by TRAP but not by BALP. In conclusion, our data indicate that both BALP and TRAP can alleviate the inhibitory effect of OPN on mineralization, suggesting a potential role for TRAP in skeletal mineralization. Further studies are warranted to explore the possible physiological relevance of TRAP in bone mineralization.Funding Agencies|Region Ostergotland; Swedish Research Council [K2015-99X-10363-23-4]; National Institute of Dental and Craniofacial Research (NIDCR) [DE12889]; National Institute of Arthritis and Musculoskeletal Diseases (NIAMS), National Institutes of Health (NIH), USA [AR53102]</p

Tartrate-resistant acid phosphatase (TRAP/ACP5) promotes metastasis-related properties via TGFβ2/TβR and CD44 in MDA-MB-231 breast cancer cells

Abstract Background Tartrate-resistant acid phosphatase (TRAP/ACP5), a metalloenzyme that is characteristic for its expression in activated osteoclasts and in macrophages, has recently gained considerable focus as a driver of metastasis and was associated with clinically relevant parameters of cancer progression and cancer aggressiveness. Methods MDA-MB-231 breast cancer cells with different TRAP expression levels (overexpression and knockdown) were generated and characterized for protein expression and activity levels. Functional cell experiments, such as proliferation, migration and invasion assays were performed as well as global phosphoproteomic and proteomic analysis was conducted to connect molecular perturbations to the phenotypic changes. Results We identified an association between metastasis-related properties of TRAP-overexpressing MDA-MB-231 breast cancer cells and a TRAP-dependent regulation of Transforming growth factor (TGFβ) pathway proteins and Cluster of differentiation 44 (CD44). Overexpression of TRAP increased anchorage-independent and anchorage-dependent cell growth and proliferation, induced a more elongated cellular morphology and promoted cell migration and invasion. Migration was increased in the presence of the extracellular matrix (ECM) proteins osteopontin and fibronectin and the basement membrane proteins collagen IV and laminin I. TRAP-induced properties were reverted upon shRNA-mediated knockdown of TRAP or treatment with the small molecule TRAP inhibitor 5-PNA. Global phosphoproteomics and proteomics analyses identified possible substrates of TRAP phosphatase activity or signaling intermediates and outlined a TRAP-dependent regulation of proteins involved in cell adhesion and ECM organization. Upregulation of TGFβ isoform 2 (TGFβ2), TGFβ receptor type 1 (TβR1) and Mothers against decapentaplegic homolog 2 (SMAD2), as well as increased intracellular phosphorylation of CD44 were identified upon TRAP perturbation. Functional antibody-mediated blocking and chemical inhibition demonstrated that TRAP-dependent migration and proliferation is regulated via TGFβ2/TβR, whereas proliferation beyond basal levels is regulated through CD44. Conclusion Altogether, TRAP promotes metastasis-related cell properties in MDA-MB-231 breast cancer cells via TGFβ2/TβR and CD44, thereby identifying a potential signaling mechanism associated to TRAP action in breast cancer cells

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

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

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