Destrin, also called actin-depolymerizing factor (ADF), exists in resting parotid tissue as phosphorylated (inactive) and dephosphorylated (active) forms, and β-adrenergic stimulation of this tissue induces dephospho...Destrin, also called actin-depolymerizing factor (ADF), exists in resting parotid tissue as phosphorylated (inactive) and dephosphorylated (active) forms, and β-adrenergic stimulation of this tissue induces dephosphorylation of destrin. It is suggested that destrin dephosphorylation is involved in cortical F-actin disruption observed in parallel with β-agonist-induced amylase secretion. At present, the phosphorylation/dephosphorylation mechanism of destrin in parotid tissue is not known. We previously detected, in a crude rat parotid extract, a constitutively active protein kinase catalyzing phosphorylation of destrin;however, its identification has been hampered by difficulty in its enrichment. The purpose of this study was to explore a simple purification method(s) for this enzyme. To this end, we first developed a high-throughput dot-blot assay for the kinase with an anti-phosphodestrin antibody and then studied its purification by column chromatography on several media. We found that the kinase could be partially purified by sequential chromatography on DEAE-cellulose, phenyl-Sepharose, and hydroxyapatite columns. In each chromatography, however, the kinase could be eluted, at the cost of resolution, only by sharp increases in the elution power of the eluent;gradual increases in the elution power resulted in unacceptably poor recovery. We confirmed that enzymatic properties of the kinase were not basically altered during the purification. Further purification of the kinase was achieved by native polyacrylamide gel electrophoresis (PAGE), which resolved the kinase activity into two bands and separated the activity from most proteins (the kinase activity after PAGE was detected with destrin-coated polyvinylidene difluoride membranes and the anti-phosphodestrin antibody). The two bands seem to constitute the major destrin-phosphorylating activity in the resting rat parotid gland. We here report its partial purification and characterization together with the detection methods.展开更多
Background: The aim of this study was to verify the efficacy of lifestyle self-monitoring for the improvement of the IBS and reveal what has been changed due to the intervention. Methods: A total of 111 nursing school...Background: The aim of this study was to verify the efficacy of lifestyle self-monitoring for the improvement of the IBS and reveal what has been changed due to the intervention. Methods: A total of 111 nursing school students were randomized into three groups, two intervention groups (a two-month intervention group, n = 34, and a four-month intervention group, n = 35) and a control group (n = 34). The intervention groups conducted lifestyle self-monitoring in conjunction with a 15-minutes group work for either two or four months. The primary outcome measure was Rome II criteria for IBS. Other outcome measures were the Hospital Anxiety and Depression Scale (HADS) and the Gastrointestinal Symptom Rating Scale (GSRS). They were assessed at the baseline and the end of both of the intervention periods. Analysis was conducted as intention-to-treat. Results: The prevalence of IBS did not change significantly after the intervention in any of the groups. The HAD-A score, a subscale of the HADS score for anxiety, decreased 1.4 points in the two-month intervention group (p = 0.02) and 2.3 points in the four-month intervention group of (p = 0.01) after intervention. The average GSRS decreased 0.2 points in the control group (p = 0.05) and 0.3 points in the four-month intervention group (p < 0.01). Conclusions: Lifestyle self-monitoring for two or four months did not reduce the prevalence of the IBS significantly, but it did decrease anxiety and improved the QOL related to gastrointestinal symptoms in female nursing school students.展开更多
文摘Destrin, also called actin-depolymerizing factor (ADF), exists in resting parotid tissue as phosphorylated (inactive) and dephosphorylated (active) forms, and β-adrenergic stimulation of this tissue induces dephosphorylation of destrin. It is suggested that destrin dephosphorylation is involved in cortical F-actin disruption observed in parallel with β-agonist-induced amylase secretion. At present, the phosphorylation/dephosphorylation mechanism of destrin in parotid tissue is not known. We previously detected, in a crude rat parotid extract, a constitutively active protein kinase catalyzing phosphorylation of destrin;however, its identification has been hampered by difficulty in its enrichment. The purpose of this study was to explore a simple purification method(s) for this enzyme. To this end, we first developed a high-throughput dot-blot assay for the kinase with an anti-phosphodestrin antibody and then studied its purification by column chromatography on several media. We found that the kinase could be partially purified by sequential chromatography on DEAE-cellulose, phenyl-Sepharose, and hydroxyapatite columns. In each chromatography, however, the kinase could be eluted, at the cost of resolution, only by sharp increases in the elution power of the eluent;gradual increases in the elution power resulted in unacceptably poor recovery. We confirmed that enzymatic properties of the kinase were not basically altered during the purification. Further purification of the kinase was achieved by native polyacrylamide gel electrophoresis (PAGE), which resolved the kinase activity into two bands and separated the activity from most proteins (the kinase activity after PAGE was detected with destrin-coated polyvinylidene difluoride membranes and the anti-phosphodestrin antibody). The two bands seem to constitute the major destrin-phosphorylating activity in the resting rat parotid gland. We here report its partial purification and characterization together with the detection methods.
文摘Background: The aim of this study was to verify the efficacy of lifestyle self-monitoring for the improvement of the IBS and reveal what has been changed due to the intervention. Methods: A total of 111 nursing school students were randomized into three groups, two intervention groups (a two-month intervention group, n = 34, and a four-month intervention group, n = 35) and a control group (n = 34). The intervention groups conducted lifestyle self-monitoring in conjunction with a 15-minutes group work for either two or four months. The primary outcome measure was Rome II criteria for IBS. Other outcome measures were the Hospital Anxiety and Depression Scale (HADS) and the Gastrointestinal Symptom Rating Scale (GSRS). They were assessed at the baseline and the end of both of the intervention periods. Analysis was conducted as intention-to-treat. Results: The prevalence of IBS did not change significantly after the intervention in any of the groups. The HAD-A score, a subscale of the HADS score for anxiety, decreased 1.4 points in the two-month intervention group (p = 0.02) and 2.3 points in the four-month intervention group of (p = 0.01) after intervention. The average GSRS decreased 0.2 points in the control group (p = 0.05) and 0.3 points in the four-month intervention group (p < 0.01). Conclusions: Lifestyle self-monitoring for two or four months did not reduce the prevalence of the IBS significantly, but it did decrease anxiety and improved the QOL related to gastrointestinal symptoms in female nursing school students.
