Kyosuke Takeshita

Background: The significance of cell population data (CPD) and leukocyte scattergrams in COVID-19 has not been fully established, partly due to the absence of serial leukocyte monitoring before and after SARS-CoV-2 infection. This study first examined changes in these parameters in non-immunosuppressed subjects over the course of infection. Subsequently, these findings were compared with those observed in patients who were immunosuppressed to assess the impact of immunosuppression. Methods: In total, 48 patients with COVID-19 were analyzed. Complete blood count (CBC) results and CPD were assessed using a Sysmex XN-9000 hematological analyzer. Results: The control and IST groups had similar clinical characteristics regarding COVID-19 severity and baseline CBC and CPD. WBC and neutrophil counts showed no significant changes immediately post onset; however, they decreased in the control group and increased in the IST group. Platelet counts decreased transiently on days 3–5 in both groups. The control group’s lymphocyte counts significantly dropped, but their lymphocyte-related CPD remained unchanged. The IST group experienced delayed lymphocyte recovery and showed reduced DNA/RNA content and cell size diversity. Scattergrams immediately after onset showed an increase in lymphocyte clusters, particularly juvenile lymphocytes, in the control group, while they decreased in the IST group. In the control group, mature neutrophils decreased while immature neutrophils increased. Conversely, the percentage of mature neutrophils increased in the IST group. Both groups showed minimal plasmacytoid lymphocyte clusters after onset. Conclusions: Immunosuppression impairs juvenile cell mobilization, which may increase susceptibility to viral impacts and potentially worsen prognosis by increasing the risk of infection.

線溶抑制因子（plasminogen activator inhibitor-1: PAI-1）は，血栓症の原因となるだけでなく，細胞外マトリクスの代謝を抑制し，組織線維化を促進する．ホルモン，炎症関連シグナル，活性酸素によって誘導され，慢性炎症を伴う疾患において重要な役割を果たす．メタボリック症候群や慢性的ストレスは，脂肪融解に伴う遊離脂肪酸の増加を介して炎症性アディポカインを増加させ，慢性炎症とPAI-1の産生を促進し，血栓傾向を高める．慢性腎臓病において，PAI-1はレニン-アンギオテンシン系の活性化や尿毒素の蓄積によって誘導され，疾患の進行を促進し，血栓傾向と心血管病の発症・増悪に関与する．老化においてPAI-1は老化関連分泌現象（Senescence-associated Secretory Phenotype: SASP）因子の一つであり，バイオマーカーおよび治療標的として注目されている．早期老化モデルklothoマウスではPAI-1の増加が老化の病態の悪化と関連し，その抑制により症状の改善が確認されている．慢性炎症によるPAI-1の増加が血栓傾向や線維化を促進し，個体の老化を加速させる可能性があり，その制御は健康寿命の延伸につながると考えられる．

Objectives: There are currently no appropriate markers and target for prophylaxis against COVID-19-related thrombosis, especially in the not-severe cases. We tested the hypothesis that inflammation is a suitable marker and target for prophylaxis against COVID-19-related thrombosis.

In patients with chronic kidney diseases (CKD), high serum indoxyl sulfate (IS) levels correlate with cardiac fibrosis and hypertrophy and thus a critical risk factor for heart failure. The aim of this study was to determine the effects of IS on cardiac function and inflammasome pathway in a rat model of CKD. We assessed the physiological and pathological changes and measured biomarkers of fibrosis and hypertrophy in the hearts of Dahl salt-sensitive (DS), DS hypertensive (DH), and DH IS-treated rats (DH + IS). Low left ventricular (LV) ejection fraction, LV dilatation, and advanced myocardial fibrosis and hypertrophy were observed in DH + IS, which resemble changes found in uremic cardiomyopathy. These changes were independent of renal function and blood pressure. RT-PCR and western blotting analysis showed upregulation of fibrosis and hypertrophy-related biomarkers and adhesion molecules in the hearts of DH + IS rats. IS activated aryl hydrocarbon receptor (AHR) pathway, nuclear factor kappa B p65 (NF-κB p65), and inflammasome in the myocardium of DH + IS rat. Moreover, IS upregulated the expression of critical NLRP3 inflammasome components (NLRP3, ASC, and procaspase-1) and increased production of IL-1β and IL-18. Finally, IS upregulated various inflammatory cytokines, such as MCP-1, TNF-α, IL-6, and TGFβ1, in the myocardium. Our results suggested that IS induced cardiac fibrosis and hypertrophy and impaired LV function through activation of cardiac NLRP3 inflammasome via the AHR/NF-κB pathway.

Notch signaling has been recognized recently as a key regulator of metabolism. Here, we determined the role of Notch1 in adipogenesis in wild-type (WT) and Notch1 hetero-mutant (N1+/−) mice provided with 12-week normal or high-fat diet. Haploinsufficiency of Notch1 was associated with adipose tissue accumulation despite similar food intake. White adipose tissue (WAT) of N1+/− showed accumulation of adipogenic cells (CD34+CD68+ cells), crown-like structures, and upregulation of cell proliferation markers (cyclin D1 and Ki67). Notch1 expression in WAT reached peak levels in 8-week-old WT mice in parallel with fat accumulation, especially under HF/HS-feeding, whereas such increment was blunted in N1+/− mice. Downstream of Notch1 haploinsufficiency, over-expression of adipogenic factors PPARγ and C/EBPα was noted following down-regulation of downstream transcriptional factors of Notch signaling (Hes-1, Pref-1, and Sox9). Both pharmacological Notch signal inhibition and Notch1 knockdown enhanced adipogenesis of 3T3-L1 preadipocytes. N1+/− mice showed impaired glucose and insulin tolerance with downregulation of IRS-1 and GLUT4 in WAT after high-fat diet. Taken together, our results suggest that haploinsufficiency of Notch1 promotes fat accumulation and adipogenesis and provides a mechanistic link between Notch signaling and development of metabolic syndrome.

Notch signaling has been recognized recently as a key regulator of metabolism. Here, we determined the role of Notch1 in adipogenesis in wild-type (WT) and Notch1 hetero-mutant (N1+/-) mice provided with 12-week normal or high-fat diet. Haploinsufficiency of Notch1 was associated with adipose tissue accumulation despite similar food intake. White adipose tissue (WAT) of N1+/- showed accumulation of adipogenic cells (CD34 + CD68 + cells), crown-like structures, and upregulation of cell proliferation markers (cyclin D1 and Ki67). Notch1 expression in WAT reached peak levels in 8-week-old WT mice in parallel with fat accumulation, especially under HF/HS-feeding, whereas such increment was blunted in N1+/- mice. Downstream of Notch1 haploinsufficiency, over-expression of adipogenic factors PPARγ and C/EBPα was noted following down-regulation of downstream transcriptional factors of Notch signaling (Hes-1, Pref-1, and Sox9). Both pharmacological Notch signal inhibition and Notch1 knockdown enhanced adipogenesis of 3T3-L1 preadipocytes. N1+/- mice showed impaired glucose and insulin tolerance with downregulation of IRS-1 and GLUT4 in WAT after high-fat diet. Taken together, our results suggest that haploinsufficiency of Notch1 promotes fat accumulation and adipogenesis and provides a mechanistic link between Notch signaling and development of metabolic syndrome.

Introduction: The aim of this study was to determine the role of Notch in indoxyl sulfate (IS)-induced vascular calcification (VC).

Stress is a pivotal factor for inflammation, reactive oxygen species (ROS) production and formation of visceral hypersensitivity (VH) in the process of gastroesophageal reflux disease (GERD). In the present study, the effects of stress on esophageal inflammation, oxidative stress and VH were investigated in a chronic restraint stress mouse model. C57BL/6J male mice were subjected to 2 weeks of intermittent restraint stress, and histopathological analysis revealed that stress induced esophageal inflammation and fibrosis, while no distinct changes were detected in non-stressed control mice. In addition, increased NADPH oxidase 4 expression was observed in the plasma and esophagus of stressed mice, indicating accumulation of ROS. The expression levels of antioxidants, including Mn-superoxide dismutase (MnSOD), Cu/Zn-SOD, catalase and glutathione peroxidase, were also analyzed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). In addition, transient receptor potential vanilloid 1 (TRPV-1) and protease-activated receptor 2 (PAR-2), which are crucial receptors for VH, were measured by immunohistochemistry and RT-qPCR. The results demonstrated that stress markedly reduced antioxidant expression, while it significantly upregulated TRPV-1 and PAR-2 expression levels in the mouse esophagus. Finally, 2 weeks of restraint stress significantly increased the esophageal and plasma levels of inflammatory cytokines, including interleukin (IL)-6, IL-8, interferon-gamma and tumor necrosis factor-alpha. Taken together, the present study results indicated that stress-induced esophageal inflammation and ROS generation involves VH.

Introduction: There is general interest in finding clinical markers for left ventricular diastolic dysfunction (LVDD), a major cause of cardiorenal syndrome leading to heart failure in chronic kidney disease (CKD) patients. The aim was to assess the utility of computed tomography (CT)-based abdominal aortic calcification (AAC) for the prediction of LVDD and prognosis of asymptomatic pre-dialysis CKD patients.