A qualitative study on family carers views on how end-of-life communication contributes to palliative oriented care in nursing home.

Background. Although family-centered communication about end-of-life care has been recognized to promote palliative-oriented care in nursing home (NH), how this communication may work is still unknown. Therefore, we explored the mechanisms by which end-of-life communication may contribute to palliative-oriented care in NH from the perspective of bereaved family carers.Methods. A descriptive qualitative design was performed. Interviews were conducted with 32 bereaved family carers whose relative had died between 45 days to 9 months prior from 13 different NHs. A two-steps analysis process firstly with deductive and then with inductive content analysis was adopted.Results. Four mechanisms by which end-of-life communication contributed to palliative-oriented care were identified: a) promoting family carers understanding about their relative's health conditions, prognosis, and treatments available; b) fostering shared decision-making between healthcare professionals and residents/family carers; c) improving knowledge of residents' preferences; and d) improving knowledge of family carers' preferences.Conclusion. Clear and in-depth communication provides insight into residents' and family carers' preferences for care and treatment at the end-of-life, and increases understanding and shared decision-making

Band 3 is an anchor protein and a target for SHP-2 tyrosine phosphatase in human erythrocytes

AbstractTyr phosphorylation of the multifunctional transmembrane protein band 3 has been implicated in several erythrocyte functions and disorders. We previously demonstrated that pervanadate treatment of human erythrocytes induces band-3 Tyr phosphorylation, which is catalyzed by the sequential action of tyrosine kinase Syk and tyrosine kinase(s) belonging to the Src family. In this study, we show that Tyr phosphorylation of band 3, elicited by pervanadate, N-ethylmaleimide, or diamide, greatly increases band-3 interaction with the tyrosine phosphatase SHP-2 in parallel with the translocation of SHP-2 to erythrocyte membranes. These events seem to be mediated by Src-like catalyzed phosphorylation of band 3 because both SHP-2 translocation to cellular membranes and its interaction with Tyr-phosphorylated protein are greatly counteracted by PP2, a specific inhibitor of Src kinases. Binding-competition experiments demonstrate that SHP-2 recruitment to band 3 occurs via its SH2 domain(s). In particular, our data support the view that SHP-2 docks specifically with P-Y359 of band 3. Experiments performed with intact erythrocytes in the presence of the SHP-2 inhibitor calpeptin suggest that, once recruited to Tyr-phosphorylated band 3, the tyrosine phosphatase dephosphorylates the protein. P-Y8, 21, and 904 are the residues affected by SHP-2, as judged by 32P-peptide mapping of band 3 digested with trypsin. These results indicate that in treated erythrocytes, recruitment of cytosolic SHP-2 to band 3 is a prerequisite for the subsequent dephosphorylation of the transmembrane protein

Phosphorylation of casein by mitochondrial protein kinase(s).

At least two protein kinase activities are bound to the rat liver mitochondrial membranes. Both activities are found to phosphorylate, besides endogenous proteins tightly bound to the membrane structures, also exogenous phosphoproteins such as casein and phosvitin. However one is able to phosphorylate both casein-bound serine and threonine residues, while the other is phosphorylating almost only serine residues

Comparative Characterization of Membrane-associated and Cytosolic Tyr-protein Kinases In Human-erythrocytes

In recent years, two protein-tyrosine kinase activities, phosphorylating tyrosine residues on the transmembrane band-3 protein, have been isolated from human erythrocyte membranes and partially characterized by different laboratories, i.e. one extracted by non-ionic detergent (Triton X-100 or Nonidet P-40), the other solubilized by 0.25 M NaCl from the detergent-insoluble residue. The present paper shows that these two membrane-associated Tyr-protein kinases purified, in the presence of bovine serum albumin, by phosphocellulose chromatography followed by heparin-Sepharose chromatography, have the same apparent molecular mass (36 kDa) determined by Ultrogel Ac44 filtration. Moreover, both Tyr-protein kinases exhibit several identical properties, including Km values for band 3, the random acidic copolymer poly(Glu,Tyr)4:1 and angiotensin II, pH dependence, response to Mn2+ and Mg2+, response to NaCl and 2,3-bisphosphoglycerate. All these properties are identical or very similar to those exhibited by the Tyr-protein kinase previously isolated by us from human erythrocyte cytosol. These results suggest that the two membrane-associated and the cytosolic Tyr-protein kinase activities are mediated by the same enzyme, distributed between the cytosol and the membrane structures

Further purification and characterization of casein kinases from human erythrocyte hemolysate. Effect of Triton X-100.

Two cyclic AMP-independent casein kinases can be isolated from human erythrocyte hemolysate, one of which (referred to as 'casein kinase S') phosphorylates only serine residues of whole commercial casein, while the other (referred to as 'casein kinase TS') phosphorylates both serine and threonine residues of the same substrate. Moreover, the casein kinase S, unlike casein kinase TS, is able to phosphorylate the erythrocyte membrane proteins. The present paper deals with the further characterization of casein kinase S, freed from histone kinase activity by DEAE and subsequent phosphocellulose chromatography of the crude hemolysate in the presence of 0.2% Triton X-100. In particular, cytosol casein kinase S exhibits some physico-chemical and catalytic properties identical to those of the membrane-bound casein kinase, solubilised and purified as previously described. Both casein kinases display the same chromatographic behaviour, the same Sepharose elution volume, the same optimal pH range, the same Km for casein and ATP, the same response to NaCl, MgCl2 and CaCl2, and the same ability to phosphorylate serine but not threonine residues of beta-casein

Phosphorylation of Membrane-proteins by Cytosolic Casein Kinases In Human-erythrocytes - Effect of Mono-valent Ions, 2,3-bisphosphoglycerate and Spermine

Membrane proteins of human erythrocytes can be phosphorylated not only by membrane casein kinase (MS) but also by cytosolic casein kinases CS and CTS, resembling casein kinase I and II, respectively. Casein kinase CS, like membrane casein kinase MS, preferentially phosphorylates membrane proteins such as band 2 (spectrin, beta-subunit) and band 3, which are the major phosphate-acceptor proteins in the endogenous phosphorylation of isolated ghosts in the presence of [gamma-32P]ATP. By contrast, cytosolic casein kinase CTS phosphorylates, in addition to band 2, some membrane proteins, whose endogenous phosphorylation in isolated ghosts under the same conditions is negligible, if any. The CS- and CTS-catalyzed phosphorylations exhibit different response to increasing NaCl (or KCl) concentrations up to physiological levels (140 mM KCl, 20 mM NaCl); i.e. CS- and MS-catalyzed phosphorylations are strongly inhibited by 75-150 mM KCl (or NaCl), while CTS-catalyzed phosphorylation is practically unaffected. In the absence of added NaCl, CS- and MS-catalyzed phosphorylations are markedly inhibited by 1.5-3 mM 2,3-bisphosphoglycerate, whereas CTS-catalyzed phosphorylation appears to be practically unaffected. Finally, CS- and MS-catalyzed phosphorylations are slightly inhibited also by 1 mM spermine, while CTS-catalyzed phosphorylation is enhanced by this polycation concentration

Phosphorylation of Cytosolic Proteins By Casein Kinases In Human-erythrocytes - Response To Ionic-strength and To 2,3-bisphosphoglycerate

The endogenous phosphorylation of human erythrocyte cytosolic proteins is markedly increased when the crude cytosol, prior to incubation in the presence of [y-32P] ATP, is submitted to DEAE-cellulose chromatography. Some proteins, including 22 and 23 kDa proteins, are preferentially phosphorylated by cytosolic casein kinase CS, whereas other proteins, including 42 kDa protein, are preferentially phosphorylated by casein kinase CTS. The CS-catalyzed phosphorylation is strongly inhibited by physiological ionic strength (150 mM KCl or NaCl) and by physiological levels (3 mM) of 2,3-bisphosphoglycerate, while CTS-catalyzed phosphorylation is unaffected. The very poor endogenous phosphorylation of these proteins in the crude cytosol may be due to the presence of other cytosolic inhibitors which are removed by DEAE-cellulose chromatography