Background. Intermediate outcomes are associated with the survival of long-term haemodialysis pat... more Background. Intermediate outcomes are associated with the survival of long-term haemodialysis patients; however, outcome variability across centres may result in heterogeneous quality of care. The study aim was to evaluate a multifaceted quality improvement activity (QIA) targeting several haemodialysis clinical performance measures. Methods. A total (prevalent and incident) of 313 patients from four dialysis units were included. The QIA was based on a multifaceted strategy involving collection of haemodialysis clinical performance measures every 6-8 months, feedback about results, improvement plans and benchmarking, and it was tested in a 3-year prospective interventional study. Two timepoints of clinical performance measures were considered for evaluating the QIA: baseline (February 2003, pre-QIA) and final (February 2006, post-QIA). Results. Centres showed significant improvement in percentage of patients with haemoglobin <11 g/dl, mean haemoglobin; percentage of patients with Kt/v <1.2, mean Kt/v; percentage of patients with phosphorous >5.5 mg/dl, mean phosphorous; percentage of patients with calcium phosphate product >55, mean calcium phosphate product; and percentage of patients with ferritin <200 ng/ml, mean ferritin. No change was observed in percentage of patients with haemoglobin between 11 and 13 g/dl, erythropoietin consumed; percentage of patients with ferritin <100 ng/ml; percentage of patients with ferritin >800 ng/ml; percentage of patients with albumin <3.5 g/dl, mean albumin; or percentage of native arteriovenous fistula. The percentage of patients with haemoglobin >13 g/dl was increased. Conclusions. Quality-improvement strategies can help improve haemodialysis performance for anaemia, dialysis dose and bone metabolism. The importance of assessing patients with high haemoglobin level should be stressed.
Background. Intermediate outcomes are associated with the survival of long-term haemodialysis pat... more Background. Intermediate outcomes are associated with the survival of long-term haemodialysis patients; however, outcome variability across centres may result in heterogeneous quality of care. The study aim was to evaluate a multifaceted quality improvement activity (QIA) targeting several haemodialysis clinical performance measures. Methods. A total (prevalent and incident) of 313 patients from four dialysis units were included. The QIA was based on a multifaceted strategy involving collection of haemodialysis clinical performance measures every 6-8 months, feedback about results, improvement plans and benchmarking, and it was tested in a 3-year prospective interventional study. Two timepoints of clinical performance measures were considered for evaluating the QIA: baseline (February 2003, pre-QIA) and final (February 2006, post-QIA). Results. Centres showed significant improvement in percentage of patients with haemoglobin <11 g/dl, mean haemoglobin; percentage of patients with Kt/v <1.2, mean Kt/v; percentage of patients with phosphorous >5.5 mg/dl, mean phosphorous; percentage of patients with calcium phosphate product >55, mean calcium phosphate product; and percentage of patients with ferritin <200 ng/ml, mean ferritin. No change was observed in percentage of patients with haemoglobin between 11 and 13 g/dl, erythropoietin consumed; percentage of patients with ferritin <100 ng/ml; percentage of patients with ferritin >800 ng/ml; percentage of patients with albumin <3.5 g/dl, mean albumin; or percentage of native arteriovenous fistula. The percentage of patients with haemoglobin >13 g/dl was increased. Conclusions. Quality-improvement strategies can help improve haemodialysis performance for anaemia, dialysis dose and bone metabolism. The importance of assessing patients with high haemoglobin level should be stressed.
The A6H monoclonal antibody (mAb) recognizes a 120 000-140 000 MW antigen that is expressed at si... more The A6H monoclonal antibody (mAb) recognizes a 120 000-140 000 MW antigen that is expressed at similar densities on 85-90% of human CD4+ and CD8+ T cells and on renal cell carcinomas. The binding of the A6H mAb induced a costimulatory signal in anti-CD3 activated T cells. In the present report, we show that A6H costimulated cell proliferation and cytokine production in purified CD4 + T cells. Unexpectedly, the CD8 + T-cell subpopulation failed to respond. CD4+ T cells costimulated with the A6H mAb upregulated CD80, CD86, CD71, interleukin-2 (IL-2)Roc, IL-2RfB and IL-2Ry, while no corresponding up-regulation of these cell surface molecules was seen in CD8 + T cells. In order to investigate the nature of the A6H mAb costimulus at the transcriptional level we have examined induction of the transcription factors OCT-1, AP-1 and NF-KB which are known to be transcriptional regulators of several cytokine and cytokine receptor genes, including the IL-2 and IL-2R genes. Co-ligation of the A6H antigen and the CD3 complex induced expression of the transcription factor AP-1 in CD4+ T cells, whereas no increase in NF-KB and octamer-binding (Oct) proteins was seen compared to T cells stimulated with anti-CD3 alone. Furthermore, no induction of AP-l was seen in A6H costimulated CD8+ T cells.
In this paper the contribution of different accessory molecules to the adhesion of resting, naive... more In this paper the contribution of different accessory molecules to the adhesion of resting, naive and memory CD4+ T cells was examined utilizing a panel of CHO cell transfectants as model antigen-presenting cells (APCs). CD4+ T lymphocytes demonstrated strong adhesion to HLA-DR4 transfected CHO cells co-expressing B7, ICAM-I or LFA-3 molecules, suggesting that all three adhesion pathways is utilized by resting CD4+ cells. Monoclonal antibodies (MoAbs) against the corresponding receptors on T cells, e.g. anti-CD28, anti-LFA-1β and anti-CD2, inhibited completely T-cell adhesion to natural ligands expressed on transfected CHOcells. Pretreatment of CD4+ T cells with NKI-L16 MoAb, which interact with an activation epitope on LFA-loc chain, enhanced adhesion to ICAM-1 but not B7 or LFA-3 expressing CHO cells. Analysis of T helper-cell subsets revealed that memory T cells bound several fold stronger to ICAM-1 expressing transfectants compared to the CD4+ 45RA+ naive T cells, whereas adhesion to B7, LFA-3- and B7/LFA-3-expressing CHO cells was similar in both T-cell subsets. The kinetics of adhesion of naive and memory CD4+ T cells to ICAM-1 was rapid and similar in both subsets. The NKI-L16 MoAb multiplied several times ICAM-1-dependent adhesion in naive compared to memory cells, which enabled the naive cells to reach a similar adhesion level as memory cells. The results suggest that resting naive CD4+ T cells utilize preferentially the CD2/LFA-3 or CD28/B7 adhesion pathways upon adhesion to APCs, while memory CD4+ T cells utilize the CD2/LFA-3, CD28/B7 and LFA-l/ICAM-1 adhesion pathways. The NK.I-L16 MoAb-induced upregulation of adhesion involves an increased affinity of LFA-1 for its ligand and not a change in the number of LFA-1 molecules. This is compatible with a view that naive cells express a large number of inactive LFA-1 molecules, whereas memory cells express preferentially activated LFA-1 molecules. The inherent low number of active LFA-I molecules on naive CD4+ T cells may be important in keeping these cells in a resting state.
We have characterized the regulation of nuclear factors involved in transcriptional control of th... more We have characterized the regulation of nuclear factors involved in transcriptional control of the interleukin-2 (IL-2) promoter-enhancer activity in Jurkat T cells stimulated with superantigen presented on HLA-DR transfectants combined with the ligands LFA-3 (CD58) and B7-1 (CD80). Gel shift analyses showed that NF-AT was strongly induced in LFA-3-costimulated Jurkat T cells, suggesting that NF-AT is a key target nuclear factor for the CD2-LFA-3 pathway. Studies using HLA-DR-B7-1-LFA-3 triple transfectants showed that the LFA-3-induced NF-AT DNA binding activity was negatively regulated by B7-1 costimulation. In contrast, induction of a CD28 response complex containing only c-Rel proteins was seen after B7-1 costimulation. Both LFA-3 costimulation and B7-1 costimulation induced the AP-1 and NF-B nuclear factors.
The specific signaling connections between the mitogen-activated protein kinases (MAPK) such as c... more The specific signaling connections between the mitogen-activated protein kinases (MAPK) such as c-Jun Nterminal kinase (JNK-1) and phosphatases PP4 and M3/6, affecting the family of early nuclear factors, is complex and remains poorly understood. JNK-1 regulates cellular differentiation, apoptosis and stress responsiveness by up-regulating early nuclear factors such as c-Jun, a member of the activating protein (AP-1) family, and the Early Growth Factor (EGR-1). C-Jun, when phosphorylated by c-Jun N-terminal kinase (JNK-1) associates with c-Fos to form the AP-1 transcription factor that activates gene expression. We have investigated the regulation of the JNK-1 kinase by co-transfecting phosphatases PP4 and M3/6 in prostate cancer cell lines PC-3 and LNCaP, which have been previously stimulated with human EGF or cisplatin. Cotransfections of plasmids expressing the JNK-1 and the serine/threonine phosphatases PP4 resulted in a significant increase in JNK-1 activity in both PC3 and LNCaP cells. In contrast, co-transfection of JNK-1 with the dual specific phosphatase serine/threonine M3/6 showed only a marginal effect in JNK-1 activity. The phosphatase M3/6 also failed in blocking the induction of JNK-1 activity observed in presence of PP4. The higher activity of JNK-1 was associated with increased activities of the factors c-Jun/AP-1 and EGR-1. This suggests that JNK-1 activity in PC-3 and LNCaP cells requires not only active PP4 for stable maintenance but also suggests that the relative degree of phosphorylation of multiple cellular components is the determinant of JNK-1 stability.
The specific signaling connections between the mitogen-activated protein kinases (MAPK) such as c... more The specific signaling connections between the mitogen-activated protein kinases (MAPK) such as c-Jun Nterminal kinase (JNK-1) and phosphatases PP4 and M3/6, affecting the family of early nuclear factors, is complex and remains poorly understood. JNK-1 regulates cellular differentiation, apoptosis and stress responsiveness by up-regulating early nuclear factors such as c-Jun, a member of the activating protein (AP-1) family, and the Early Growth Factor (EGR-1). C-Jun, when phosphorylated by c-Jun N-terminal kinase (JNK-1) associates with c-Fos to form the AP-1 transcription factor that activates gene expression. We have investigated the regulation of the JNK-1 kinase by co-transfecting phosphatases PP4 and M3/6 in prostate cancer cell lines PC-3 and LNCaP, which have been previously stimulated with human EGF or cisplatin. Cotransfections of plasmids expressing the JNK-1 and the serine/threonine phosphatases PP4 resulted in a significant increase in JNK-1 activity in both PC3 and LNCaP cells. In contrast, co-transfection of JNK-1 with the dual specific phosphatase serine/threonine M3/6 showed only a marginal effect in JNK-1 activity. The phosphatase M3/6 also failed in blocking the induction of JNK-1 activity observed in presence of PP4. The higher activity of JNK-1 was associated with increased activities of the factors c-Jun/AP-1 and EGR-1. This suggests that JNK-1 activity in PC-3 and LNCaP cells requires not only active PP4 for stable maintenance but also suggests that the relative degree of phosphorylation of multiple cellular components is the determinant of JNK-1 stability.
Background. Intermediate outcomes are associated with the survival of long-term haemodialysis pat... more Background. Intermediate outcomes are associated with the survival of long-term haemodialysis patients; however, outcome variability across centres may result in heterogeneous quality of care. The study aim was to evaluate a multifaceted quality improvement activity (QIA) targeting several haemodialysis clinical performance measures. Methods. A total (prevalent and incident) of 313 patients from four dialysis units were included. The QIA was based on a multifaceted strategy involving collection of haemodialysis clinical performance measures every 6-8 months, feedback about results, improvement plans and benchmarking, and it was tested in a 3-year prospective interventional study. Two timepoints of clinical performance measures were considered for evaluating the QIA: baseline (February 2003, pre-QIA) and final (February 2006, post-QIA). Results. Centres showed significant improvement in percentage of patients with haemoglobin <11 g/dl, mean haemoglobin; percentage of patients with Kt/v <1.2, mean Kt/v; percentage of patients with phosphorous >5.5 mg/dl, mean phosphorous; percentage of patients with calcium phosphate product >55, mean calcium phosphate product; and percentage of patients with ferritin <200 ng/ml, mean ferritin. No change was observed in percentage of patients with haemoglobin between 11 and 13 g/dl, erythropoietin consumed; percentage of patients with ferritin <100 ng/ml; percentage of patients with ferritin >800 ng/ml; percentage of patients with albumin <3.5 g/dl, mean albumin; or percentage of native arteriovenous fistula. The percentage of patients with haemoglobin >13 g/dl was increased. Conclusions. Quality-improvement strategies can help improve haemodialysis performance for anaemia, dialysis dose and bone metabolism. The importance of assessing patients with high haemoglobin level should be stressed.
Background. Intermediate outcomes are associated with the survival of long-term haemodialysis pat... more Background. Intermediate outcomes are associated with the survival of long-term haemodialysis patients; however, outcome variability across centres may result in heterogeneous quality of care. The study aim was to evaluate a multifaceted quality improvement activity (QIA) targeting several haemodialysis clinical performance measures. Methods. A total (prevalent and incident) of 313 patients from four dialysis units were included. The QIA was based on a multifaceted strategy involving collection of haemodialysis clinical performance measures every 6-8 months, feedback about results, improvement plans and benchmarking, and it was tested in a 3-year prospective interventional study. Two timepoints of clinical performance measures were considered for evaluating the QIA: baseline (February 2003, pre-QIA) and final (February 2006, post-QIA). Results. Centres showed significant improvement in percentage of patients with haemoglobin <11 g/dl, mean haemoglobin; percentage of patients with Kt/v <1.2, mean Kt/v; percentage of patients with phosphorous >5.5 mg/dl, mean phosphorous; percentage of patients with calcium phosphate product >55, mean calcium phosphate product; and percentage of patients with ferritin <200 ng/ml, mean ferritin. No change was observed in percentage of patients with haemoglobin between 11 and 13 g/dl, erythropoietin consumed; percentage of patients with ferritin <100 ng/ml; percentage of patients with ferritin >800 ng/ml; percentage of patients with albumin <3.5 g/dl, mean albumin; or percentage of native arteriovenous fistula. The percentage of patients with haemoglobin >13 g/dl was increased. Conclusions. Quality-improvement strategies can help improve haemodialysis performance for anaemia, dialysis dose and bone metabolism. The importance of assessing patients with high haemoglobin level should be stressed.
The A6H monoclonal antibody (mAb) recognizes a 120 000-140 000 MW antigen that is expressed at si... more The A6H monoclonal antibody (mAb) recognizes a 120 000-140 000 MW antigen that is expressed at similar densities on 85-90% of human CD4+ and CD8+ T cells and on renal cell carcinomas. The binding of the A6H mAb induced a costimulatory signal in anti-CD3 activated T cells. In the present report, we show that A6H costimulated cell proliferation and cytokine production in purified CD4 + T cells. Unexpectedly, the CD8 + T-cell subpopulation failed to respond. CD4+ T cells costimulated with the A6H mAb upregulated CD80, CD86, CD71, interleukin-2 (IL-2)Roc, IL-2RfB and IL-2Ry, while no corresponding up-regulation of these cell surface molecules was seen in CD8 + T cells. In order to investigate the nature of the A6H mAb costimulus at the transcriptional level we have examined induction of the transcription factors OCT-1, AP-1 and NF-KB which are known to be transcriptional regulators of several cytokine and cytokine receptor genes, including the IL-2 and IL-2R genes. Co-ligation of the A6H antigen and the CD3 complex induced expression of the transcription factor AP-1 in CD4+ T cells, whereas no increase in NF-KB and octamer-binding (Oct) proteins was seen compared to T cells stimulated with anti-CD3 alone. Furthermore, no induction of AP-l was seen in A6H costimulated CD8+ T cells.
In this paper the contribution of different accessory molecules to the adhesion of resting, naive... more In this paper the contribution of different accessory molecules to the adhesion of resting, naive and memory CD4+ T cells was examined utilizing a panel of CHO cell transfectants as model antigen-presenting cells (APCs). CD4+ T lymphocytes demonstrated strong adhesion to HLA-DR4 transfected CHO cells co-expressing B7, ICAM-I or LFA-3 molecules, suggesting that all three adhesion pathways is utilized by resting CD4+ cells. Monoclonal antibodies (MoAbs) against the corresponding receptors on T cells, e.g. anti-CD28, anti-LFA-1β and anti-CD2, inhibited completely T-cell adhesion to natural ligands expressed on transfected CHOcells. Pretreatment of CD4+ T cells with NKI-L16 MoAb, which interact with an activation epitope on LFA-loc chain, enhanced adhesion to ICAM-1 but not B7 or LFA-3 expressing CHO cells. Analysis of T helper-cell subsets revealed that memory T cells bound several fold stronger to ICAM-1 expressing transfectants compared to the CD4+ 45RA+ naive T cells, whereas adhesion to B7, LFA-3- and B7/LFA-3-expressing CHO cells was similar in both T-cell subsets. The kinetics of adhesion of naive and memory CD4+ T cells to ICAM-1 was rapid and similar in both subsets. The NKI-L16 MoAb multiplied several times ICAM-1-dependent adhesion in naive compared to memory cells, which enabled the naive cells to reach a similar adhesion level as memory cells. The results suggest that resting naive CD4+ T cells utilize preferentially the CD2/LFA-3 or CD28/B7 adhesion pathways upon adhesion to APCs, while memory CD4+ T cells utilize the CD2/LFA-3, CD28/B7 and LFA-l/ICAM-1 adhesion pathways. The NK.I-L16 MoAb-induced upregulation of adhesion involves an increased affinity of LFA-1 for its ligand and not a change in the number of LFA-1 molecules. This is compatible with a view that naive cells express a large number of inactive LFA-1 molecules, whereas memory cells express preferentially activated LFA-1 molecules. The inherent low number of active LFA-I molecules on naive CD4+ T cells may be important in keeping these cells in a resting state.
We have characterized the regulation of nuclear factors involved in transcriptional control of th... more We have characterized the regulation of nuclear factors involved in transcriptional control of the interleukin-2 (IL-2) promoter-enhancer activity in Jurkat T cells stimulated with superantigen presented on HLA-DR transfectants combined with the ligands LFA-3 (CD58) and B7-1 (CD80). Gel shift analyses showed that NF-AT was strongly induced in LFA-3-costimulated Jurkat T cells, suggesting that NF-AT is a key target nuclear factor for the CD2-LFA-3 pathway. Studies using HLA-DR-B7-1-LFA-3 triple transfectants showed that the LFA-3-induced NF-AT DNA binding activity was negatively regulated by B7-1 costimulation. In contrast, induction of a CD28 response complex containing only c-Rel proteins was seen after B7-1 costimulation. Both LFA-3 costimulation and B7-1 costimulation induced the AP-1 and NF-B nuclear factors.
The specific signaling connections between the mitogen-activated protein kinases (MAPK) such as c... more The specific signaling connections between the mitogen-activated protein kinases (MAPK) such as c-Jun Nterminal kinase (JNK-1) and phosphatases PP4 and M3/6, affecting the family of early nuclear factors, is complex and remains poorly understood. JNK-1 regulates cellular differentiation, apoptosis and stress responsiveness by up-regulating early nuclear factors such as c-Jun, a member of the activating protein (AP-1) family, and the Early Growth Factor (EGR-1). C-Jun, when phosphorylated by c-Jun N-terminal kinase (JNK-1) associates with c-Fos to form the AP-1 transcription factor that activates gene expression. We have investigated the regulation of the JNK-1 kinase by co-transfecting phosphatases PP4 and M3/6 in prostate cancer cell lines PC-3 and LNCaP, which have been previously stimulated with human EGF or cisplatin. Cotransfections of plasmids expressing the JNK-1 and the serine/threonine phosphatases PP4 resulted in a significant increase in JNK-1 activity in both PC3 and LNCaP cells. In contrast, co-transfection of JNK-1 with the dual specific phosphatase serine/threonine M3/6 showed only a marginal effect in JNK-1 activity. The phosphatase M3/6 also failed in blocking the induction of JNK-1 activity observed in presence of PP4. The higher activity of JNK-1 was associated with increased activities of the factors c-Jun/AP-1 and EGR-1. This suggests that JNK-1 activity in PC-3 and LNCaP cells requires not only active PP4 for stable maintenance but also suggests that the relative degree of phosphorylation of multiple cellular components is the determinant of JNK-1 stability.
The specific signaling connections between the mitogen-activated protein kinases (MAPK) such as c... more The specific signaling connections between the mitogen-activated protein kinases (MAPK) such as c-Jun Nterminal kinase (JNK-1) and phosphatases PP4 and M3/6, affecting the family of early nuclear factors, is complex and remains poorly understood. JNK-1 regulates cellular differentiation, apoptosis and stress responsiveness by up-regulating early nuclear factors such as c-Jun, a member of the activating protein (AP-1) family, and the Early Growth Factor (EGR-1). C-Jun, when phosphorylated by c-Jun N-terminal kinase (JNK-1) associates with c-Fos to form the AP-1 transcription factor that activates gene expression. We have investigated the regulation of the JNK-1 kinase by co-transfecting phosphatases PP4 and M3/6 in prostate cancer cell lines PC-3 and LNCaP, which have been previously stimulated with human EGF or cisplatin. Cotransfections of plasmids expressing the JNK-1 and the serine/threonine phosphatases PP4 resulted in a significant increase in JNK-1 activity in both PC3 and LNCaP cells. In contrast, co-transfection of JNK-1 with the dual specific phosphatase serine/threonine M3/6 showed only a marginal effect in JNK-1 activity. The phosphatase M3/6 also failed in blocking the induction of JNK-1 activity observed in presence of PP4. The higher activity of JNK-1 was associated with increased activities of the factors c-Jun/AP-1 and EGR-1. This suggests that JNK-1 activity in PC-3 and LNCaP cells requires not only active PP4 for stable maintenance but also suggests that the relative degree of phosphorylation of multiple cellular components is the determinant of JNK-1 stability.
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