Identification and Biochemical Characterization of High Mobility Group Protein 20A as a Novel Ca2+/S100A6 Target

During screening of protein-protein interactions, using human protein arrays carrying 19,676 recombinant glutathione s-transferase (GST)-fused human proteins, we identified the high-mobility protein group 20A (HMG20A) as a novel S100A6 binding partner. We confirmed the Ca2+-dependent interaction of HMG20A with S100A6 by the protein array method, biotinylated S100A6 overlay, and GST-pulldown assay in vitro and in transfected COS-7 cells. Co-immunoprecipitation of S100A6 with HMG20A from HeLa cells in a Ca2+-dependent manner revealed the physiological relevance of the S100A6/HMG20A interaction. In addition, HMG20A has the ability to interact with S100A1, S100A2, and S100B in a Ca2+-dependent manner, but not with S100A4, A11, A12, and calmodulin. S100A6 binding experiments using various HMG20A mutants revealed that Ca2+/S100A6 interacts with the C-terminal region (residues 311-342) of HMG20A with stoichiometric binding (HMG20A:S100A6 dimer = 1:1). This was confirmed by the fact that a GST-HMG20A mutant lacking the S100A6 binding region (residues 311-347, HMG20A-Delta C) failed to interact with endogenous S100A6 in transfected COS-7 cells, unlike wild-type HMG20A. Taken together, these results identify, for the first time, HMG20A as a target of Ca2+/S100 proteins, and may suggest a novel linkage between Ca2+/S100 protein signaling and HMG20A function, including in the regulation of neural differentiation

Regulation of the tubulin polymerization-promoting protein by Ca2+/S100 proteins

To elucidate S100 protein-mediated signaling pathways, we attempted to identify novel binding partners for S100A2 by screening protein arrays carrying 19,676 recombinant glutathione S-transferase (GST)-fused human proteins with biotinylated S100A2. Among newly discovered putative S100A2 interactants, including TMLHE, TRH, RPL36, MRPS34, CDR2L, OIP5, and MED29, we identified and characterized the tubulin polymerization-promoting protein (TPPP) as a novel S100A2-binding protein. We confirmed the interaction of TPPP with Ca2+/S100A2 by multiple independent methods, including the protein array method, S100A2 overlay, and pulldown assay in vitro and in transfected COS-7 cells. Based on the results from the S100A2 overlay assay using various GST-TPPP mutants, the S100A2-binding region was identified in the C-terminal (residues 111-160) of the central core domain of a monomeric form of TPPP that is involved in TPPP dimerization. Chemical cross-linking experiments indicated that S100A2 suppresses dimer formation of His-tagged TPPP in a dosedependent and a Ca2+-dependent manner. In addition to S100A2, TPPP dimerization is disrupted by other multiple S100 proteins, including S100A6 and S100B, in a Ca2+-dependent manner but not by S100A4. This is consistent with the fact that S100A6 and S100B, but not S100A4, are capable of interacting with GST-TPPP in the presence of Ca2+. Considering these results together, TPPP was identified as a novel target for S100A2, and it is a potential binding target for other multiple S100 proteins, including S100A6 and S100B. Direct binding of the S100 proteins with TPPP may cause disassembly of TPPP dimer formation in response to the increasing concentration of intracellular Ca2+, thus resulting in the regulation of the physiological function of TPPP, such as microtubule organization

A subjective well-being of the aged - psychological characteristics of elderly people of 90 or older -

高齢化社会の到来に伴い一言で老人といっても幅広い年代が対象となり、一律に老人では説明できない。実際看護をしていると90歳をすぎた老人はその年代迄にはない、穏やかさ、焦りのなさ、人生を達観しているような感じを受けることが多い。そこで90歳以上の老人にみられるイメージや心理面の特徴を明らかにし看護実践の一助としたいと考えた。方法は80歳以上の入院患者50名に身体、生活、家族面からみた現状、他者から見たイメージ、主観的幸福感を調査した。その結果、90歳以上は80歳代にくらべ看護者に肯定的イメージに受け取られる傾向にあった。主観的幸福感を示すモラール得点の総合点では差はなかったが得点する内容に差が見られ、80歳は積極的な生き方で得点しているものが多く90歳代は現状に満足している点で得点している者が多かった。看護者からみたイメージと本人の主観的幸福感は両年代とも肯定的イメージで相関した。The population of the aged is currently increasing with advanced medical science and technology. The aged have become to keep their lives long. In this paper, We especially focused on their psychological characteristics of elderly people of 90 and older to offer high quality of nursing care for them. We interviewed 50 patients of 80 years and older about a subjective well-being, using a questionaire based on Philadelphia Geriatric Center Morale Scale. And also we asked 50 nurses, who took care of them exclusively, a questinaire about their images of the aged. We report as follows : 1. The nurses estimated that their images of the second group (range : 90 years old ~) were better than the first group (range : 80~89 years old). 2. In subjective well-being which Philadelphia Geriatric Center Morale Scale showed, above-mentioned both groups were similar in total scores. However, each item was marked differently by them. 3. In both groups, their subjective well-being correlated with good images for them

Risk Factors for Restenosis after Percutaneous Coronary Intervention with Sirolimus- and Paclitaxel-eluting Stents

To identify risk factors for restenosis after percutaneous coronary intervention with sirolimus (SES)- or paclitaxel (PES)-eluting stents. The clinical outcomes of 894 patients treated with either SES (n = 462) or PES (n = 432) between January 2005 and January 2010 were evaluated. Multivariate logistic regression analysis showed that long ( > 20mm)(odds ratio [OR], 1.87; 95% confidence interval [CI], 1.07-3.33; P = 0.03) or bent (angle > 45°) lesions (OR, 2.57; 95% CI, 1.47- 4.49; P < 0.01) were independent risk factors for restenosis with SES, and that hemodialysis (OR, 7.61; 95% CI, 2.78- 20.85; P < 0.01) and long (OR, 2.63; 95% CI, 1.18-5.84; P = 0.02) or bent lesions (OR, 3.47; 95% CI, 1.65-7.27;P < 0.01) were independent risk factors for target lesion revascularization (TLR) with SES. In contrast, no independent risk factors for restenosis and TLR were found for lesions treated with PES. The rate of TLR was significantly higher in patients on hemodialysis or in those with long lesions in the SES group (hemodialysis, 30.4% vs. 11.1%, P = 0.02; long lesions, 13.2% vs. 4.4%, P < 0.01; for SES vs. PES, respectively). Rates of restenosis and TLR were significantly higher in patients with bent lesions in the SES group (restenosis, 30.8% vs. 15.6%, P < 0.01; TLR, 20.0% vs. 5.8%, P < 0.01; for SES and PES, respectively). Most clinical studies have described better angiographic results for SES compared to PES. However, PES might result in better clinical outcomes than SES for patients on hemodialysis or for those with long or bent lesions

Comparison of Mid-term Angiographic and Clinical Outcomes Following Zotarolimus-eluting Stent and Paclitaxel-eluting Stent Implantation Based on Lesion Complexity

First-generation drug-eluting stents (DESs) have reduced angiographic and clinical restenosis rates compared to bare-metal stents (BMSs). Zotarolimus-eluting stents (ZESs) are second-generation drug-eluting stents: however, the clinical efficacy of ZES implantation is unclear because late loss associated with ZESs is reportedly higher than that observed for other DESs. The aim of this study was to evaluate the clinical efficacy of ZESs compared to paclitaxel-eluting stents (PESs). We retrospectively evaluated the angiographic and clinical outcomes of 431 lesions in 342 patients treated with PESs and 153 lesions in 121 patients treated with ZESs in our hospital between May 2007 and December 2010. Follow-up angiographic examinations were performed eight months post-treatment and clinical outcomes were assessed one year after the procedure. Quantitative coronary angiographic analyses showed that late loss was significantly higher for ZESs than PESs (0.82 ± 0.73 mm vs 0.47 ± 0.68 mm; P = 0.003). However, there was no significant difference in target lesion revascularization (TLR) between the two groups (ZES: 15 lesions, 9.8% vs PES: 25 lesions, 5.8%; P = 0.092). When comparing stents according to the American College of Cardiology/American Heart Association (ACC/AHA) lesion type, the TLR rate in the ZES group was significantly lower than in the PES group (0% vs 7.0%; P = 0.038) for Type A/B1 lesions, but the TLR rate for type B2/C lesions in the ZES group was significantly higher than in the PES group (15.8% vs 5.3%; P = 0.009). Multivariate logistic regression analysis showed that dialysis (OR: 35.54; 95% CI: 3.15-400.67; P = 0.039) and pre-minimal lumen diameter (OR: 0.036; 95% CI: 0.002-0.541; P = 0.016) were independent predictors of TLR in ZES-treated lesions. However, no factors predicted TLR in PES-treated lesions. Our study demonstrated excellent outcomes with ZESs for simple lesions, but it is necessary to carefully implant ZESs in complex lesions, such as ACC/AHA type B2/C lesions

Percutaneous coronary intervention for a healed erosion with excimer laser coronary angioplasty and drug-coated balloon angioplasty: a case report

BackgroundHealed plaque, characterized by distinct layers of organizing thrombus and collagen, is the hallmark of tissue self-repair. However, the efficacy of excimer laser coronary angioplasty (ELCA) followed by drug-coated balloon (DCB) angioplasty in patients with healed plaques is not fully understood.Case summaryA 42-year-old woman with a history of anxiety disorder was admitted to our institution with worsening chest pain and subsequently diagnosed with anterior non-ST-elevation myocardial infarction. Coronary angiography revealed severe stenosis in the proximal left anterior descending artery (LAD) despite Thrombolysis in Myocardial Infarction (TIMI) grade 3. Optical coherence tomography (OCT) showed healed plaques with partial macrophage accumulation and no fresh thrombus. Plaque disruption and thin-cap fibrous atheroma were not identified in the culprit lesions. Intravascular ultrasound (IVUS) confirmed high-intensity marginal irregular masses at the culprit site, suggesting that the thrombus was formed by plaque erosion rather than lipid plaque or necrotic tissue. With lesion modification using ELCA prior to DCB angioplasty, OCT examination of the LAD after ELCA showed a significant reduction in plaque burden and preserved lumen size. Post-percutaneous coronary intervention angiography revealed no stenosis with TIMI grade 3. A follow-up coronary computed tomography scan showed no angiographic restenosis, and the patient remained symptom-free.ConclusionsHere we describe a case in which OCT and IVUS evaluation suggested organizing thrombus due to erosion healing, and a favorable outcome was achieved with the combination of ELCA and DCB. The combination use of ELCA and DCB might be a potential strategy for acute coronary syndrome patients with organizing thrombus

Impact of Native Coronary Artery Calcification on Lesion Outcome Following Drug-Coated Balloon Angioplasty for Treatment of In-Stent Restenosis

This study aimed to clarify whether native coronary artery（CA） calcification before index percutaneous coronary intervention（PCI） has an impact on the effectiveness of drug-coated balloon（DCB） angioplasty for the treatment of in-stent restenosis（ISR）. 100consecutive patients with 166ISR lesions underwent quantitative coronary angiography（QCA） before and after index PCI and before and after DCB angioplasty for ISR. CA calcification before index PCI was assessed by angiography and results were analyzed to reveal the predictive values for target lesion revascularization（TLR） and major adverse cardiac events（MACE）. During 1.03±1.03years of follow-up, TLR occurred in 44lesions（26.5%） and MACE in 33 patients（33%）. On multivariate analysis, CA calcification before index PCI（p=0.016）, and % diameter of stenosis（%DS）≥73%（p=0.023） and minimal lumen diameter（MLD）<0.65mm（p=0.001） before DCB angioplasty were independent predictors for TLR after DCB angioplasty. MACE was also associated with CA calcification before index PCI（p=0.01）, and %DS≥73%（p=0.001） and MLD<0.65mm（p=0.01） before DCB angioplasty, but only %DS≥73% before DCB angioplasty was an independent predictor for MACE after DCB angioplasty（p=0.039）. The combination of CA calcification before index PCI and these QCA factors before DCB angioplasty was an independent and more powerful predictor for MACE than the QCA factors alone（p<0.001）. Thereafter, the combination of CA calcification and %DS≥73% before DCB angioplasty stratified the risk of MACE after DCB angioplasty（p<0.05）. CA calcification before index PCI, as well as anatomical information at ISR, have an impact on outcome after DCB angioplasty for ISR

CD153/CD30 signaling promotes age-dependent tertiary lymphoid tissue expansion and kidney injury

高齢者腎臓病を悪化させる原因細胞・分子の同定に成功. 京都大学プレスリリース. 2021-11-30.A new drug target for kidney disease. 京都大学プレスリリース. 2021-11-30.Tertiary lymphoid tissues (TLTs) facilitate local T- and B-cell interactions in chronically inflamed organs. However, the cells and molecular pathways that govern TLT formation are poorly defined. Here we identify TNF superfamily CD153-CD30 signaling between two unique age-dependent lymphocyte subpopulations, CD153⁺PD-1⁺CD4⁺ senescence-associated T (SAT) cells and CD30+T-bet+ age-associated B cells (ABCs), as a driver for TLT expansion. SAT cells, which produced ABC-inducing factors IL21 and IFNγ, and ABCs progressively accumulated within TLTs in aged kidneys after injury. Notably, in kidney injury models, CD153 or CD30 deficiency impaired functional SAT cell induction, which resulted in reduced ABC numbers and attenuated TLT formation with improved inflammation, fibrosis and renal function. Attenuated TLT formation after transplantation of CD153-deficient bone marrow further supported the importance of CD153 in immune cells. Clonal analysis revealed that SAT cells and ABCs in the kidneys arose from both local differentiation and recruitment from the spleen. In the synovium of aged rheumatoid arthritis patients, T peripheral helper/T follicular helper cells and ABCs also expressed CD153 and CD30, respectively. Together, our data reveal a previously unappreciated function of CD153-CD30 signaling in TLT formation and propose targeting CD153-CD30 signaling pathway as a therapeutic target for slowing kidney disease progression

Significance of Coronary Artery Calcium Score in the Target Lesion Evaluated by Multi-detector Computed Tomography for Selecting Treatment of Rotational Atherectomy in Patients with Coronary Artery Disease

We investigated whether coronary artery calcium score (CAC) in the target lesion on the multidetector computed tomography angiography (CTA) predicts the addition of the Rotational atherectomy (Rota) during percutaneous coronary intervention (PCI). Lesion CAC on CTA were evaluated with quantitative coronary analysis (QCA) on coronary angiography for predicting the Rota treatment in 114 consecutive patients (165 target lesions) with first PCI (68 ± 9 years old, females: 17.6%). Rota was added in 8 patients (11 lesions). The lesion length and diameter stenosis on QCA, and lesion length and lesion CAC on CTA were the primary factors associated with the addition of Rota. Using the cut-off value based on receiver operating characteristic analysis, the sensitivity and specificity for predicting the Rota based on QCA was 72.7% in 8 of 11 lesions (vessels) with Rota and the specificity was 74% in 114 of 154 without Rota in the lesion length of ≥ 23mm (χ2=10.9, p=0.001), and 54.5% in 6 of 11 lesions with Rota and the specificity was 79.2% in 122 of 154 without Rota in the diameter stenosis of ≥ 83% (χ2=6.6, p=0.01). Those based on CTA were 90.9% in 10 of 11 lesions with Rota and 77.3% in 119 of 154 without Rota in the lesion length of ≥ 34mm (χ2=24.1, p<0.001), and 90.9% in 10 of 11 with Rota and 88.3% in 136 of 154 without Rota in the lesions with CAC ≥453 (χ2=45.7, p<0.001). Lesion CAC on CTA is most predictive of addition of Rota during PCI

Functional Characterization of FLT3 Receptor Signaling Deregulation in Acute Myeloid Leukemia by Single Cell Network Profiling (SCNP)

Molecular characterization of the FMS-like tyrosine kinase 3 receptor (FLT3) in cytogenetically normal acute myeloid leukemia (AML) has recently been incorporated into clinical guidelines based on correlations between FLT3 internal tandem duplications (FLT3-ITD) and decreased disease-free and overall survival. These mutations result in constitutive activation of FLT3, and FLT3 inhibitors are currently undergoing trials in AML patients selected on FLT3 molecular status. However, the transient and partial responses observed suggest that FLT3 mutational status alone does not provide complete information on FLT3 biological activity at the individual patient level. Examination of variation in cellular responsiveness to signaling modulation may be more informative.Using single cell network profiling (SCNP), cells were treated with extracellular modulators and their functional responses were quantified by multiparametric flow cytometry. Intracellular signaling responses were compared between healthy bone marrow myeloblasts (BMMb) and AML leukemic blasts characterized as FLT3 wild type (FLT3-WT) or FLT3-ITD. Compared to healthy BMMb, FLT3-WT leukemic blasts demonstrated a wide range of signaling responses to FLT3 ligand (FLT3L), including elevated and sustained PI3K and Ras/Raf/Erk signaling. Distinct signaling and apoptosis profiles were observed in FLT3-WT and FLT3-ITD AML samples, with more uniform signaling observed in FLT3-ITD AML samples. Specifically, increased basal p-Stat5 levels, decreased FLT3L induced activation of the PI3K and Ras/Raf/Erk pathways, decreased IL-27 induced activation of the Jak/Stat pathway, and heightened apoptotic responses to agents inducing DNA damage were observed in FLT3-ITD AML samples. Preliminary analysis correlating these findings with clinical outcomes suggests that classification of patient samples based on signaling profiles may more accurately reflect FLT3 signaling deregulation and provide additional information for disease characterization and management.These studies show the feasibility of SCNP to assess modulated intracellular signaling pathways and characterize the biology of individual AML samples in the context of genetic alterations