Myosin Light Chain Phosphatase Plays an Important Role in Cardiac Fibrosis in a Model of Mineralocorticoid Receptor-Associated Hypertension.

BACKGROUND: Myosin phosphatase targeting subunit 2 (MYPT2) is an important subunit of cardiac MLC (myosin light chain) phosphatase, which plays a crucial role in regulating the phosphorylation of MLC to phospho-MLC (p-MLC). A recent study demonstrated mineralocorticoid receptor-related hypertension is associated with RhoA/Rho-associated kinase/MYPT1 signaling upregulation in smooth muscle cells. Our purpose is to investigate the effect of MYPT2 on cardiac function and fibrosis in mineralocorticoid receptor-related hypertension. METHODS AND RESULTS: HL-1 murine cardiomyocytes were incubated with different concentrations or durations of aldosterone. After 24-hour stimulation, aldosterone increased CTGF (connective tissue growth factor) and MYPT2 and decreased p-MLC in a dose-dependent manner. MYPT2 knockdown decreased CTGF. Cardiac-specific MYPT2-knockout (c-MYPT2-/-) mice exhibited decreased type 1 phosphatase catalytic subunit ß and increased p-MLC. A disease model of mouse was induced by subcutaneous aldosterone and 8% NaCl food for 4 weeks after uninephrectomy. Blood pressure elevation and left ventricular hypertrophy were observed in both c-MYPT2-/- and MYPT2+/+ mice, with no difference in heart weights or nuclear localization of mineralocorticoid receptor in cardiomyocytes. However, c-MYPT2-/- mice had higher ejection fraction and fractional shortening on echocardiography after aldosterone treatment. Histopathology revealed less fibrosis, reduced CTGF, and increased p-MLC in c-MYPT2-/- mice. Basal global radial strain and global longitudinal strain were higher in c-MYPT2-/- than in MYPT2+/+ mice. After aldosterone treatment, both global radial strain and global longitudinal strain remained higher in c-MYPT2-/- mice compared with MYPT2+/+ mice. CONCLUSIONS: Cardiac-specific MYPT2 knockout leads to decreased myosin light chain phosphatase and increased p-MLC. MYPT2 deletion prevented cardiac fibrosis and dysfunction in a model of mineralocorticoid receptor-associated hypertension.

Prognostic Impact of Prehospital Simple Risk Index in Patients With ST-Elevation Myocardial Infarction.

BACKGROUND: The simple risk index recorded in the emergency room (ER-SRI), which is calculated using the formula (heart rate × [age / 10]2) / systolic blood pressure, was shown to be able to stratify the prognosis in ST-elevation myocardial infarction (STEMI) patients. However, the prognostic impact of the prehospital simple risk index (Pre-SRI) remains unknown.Methods and Results: This study enrolled 2,047 STEMI patients from the Mie Acute Coronary Syndrome (ACS) registry. Pre-SRI was calculated using prehospital data and ER-SRI was calculated using emergency room data. The primary endpoint was 30-day all-cause mortality. The cut-off values of Pre-SRI and ER-SRI for predicting 30-day mortality were 34.8 and 34.1, with accuracies of 0.816 and 0.826 based on receiver operating characteristic analyses (P<0.001 for both). There was no difference in the accuracy of the 2 indices. Multivariate Cox regression analysis demonstrated that a High Pre-SRI (≥34) was a significant independent predictor of 30-day mortality. With combined Pre-SRI and ER-SRI assessment, patients with High Pre-SRI/High ER-SRI showed significantly higher mortality than those with High Pre-SRI/Low ER-SRI, Low Pre-SRI/High ER-SRI, and Low Pre-SRI/Low ER-SRI (P<0.001). The addition of High Pre-SRI to High ER-SRI showed incremental prognostic value of the Pre-SRI. CONCLUSIONS: Pre-SRI can identify high-risk STEMI patients at an early stage and combined assessment with Pre-SRI and ER-SRI could be of incremental prognostic value for risk stratification in STEMI patients.

Clinical characteristics and the risk of hospitalization of patients with coronavirus disease 2019 quarantined in a designated hotel in Japan.

The aim was to investigate the clinical characteristics of coronavirus disease 2019 (COVID-19) patients who were admitted to a designated hotel, and to clarify the risk factors for hospitalization of such patients with clinical deterioration. The medical records of COVID-19 patients who were admitted to the designated hotel in Mie Prefecture, Japan, between August 2020 and September 2021 were reviewed retrospectively. Of the 1,087 COVID-19 patients who were admitted to the designated hotel, 936 patients (32.1± 12.8 years, 61.3% male) were recruited after excluding patients under the age of 15 years (n = 33), those admitted from the hospital (n = 111, 10.2%), COVID-19 vaccinated patients (n = 4, 0.4%), and those who were discharged to their own home due to social disorders (n = 3). During the study period, 884 patients (94.4%) were discharged to their own home with improving symptoms, whereas 52 patients (5.6%) were hospitalized for a deteriorating clinical condition. The logistic regression analyses showed that older age (≥ 40 years), higher body mass index (≥ 25 kg/m2), hypertension were the risk factors for hospitalization. As the new risk scale score based on the results of the odds ratios increased, the hospitalization rate increased significantly: 2.0% at 0-1 points, 9.7% at 2-3 points, and 28.8% at 4-5 points (p < 0.001). None of the 52 hospitalized patients died, and none developed serious complications from COVID-19 after hospitalization. In conclusion, the designated accommodation program for COVID-19 patients was safe, especially for those with a low risk for hospitalization.

Demographic Trends and Changes in the Pre- and In-Hospital Medical Management of Acute Myocardial Infarction During the First 12 Months of the COVID-19 Pandemic in Mie Prefecture　- Report From the Mie ACS Registry.

Background: Even though hospital admissions for acute myocardial infarction (AMI) decreased globally during the COVID-19 pandemic in early 2020, limited information is available on subsequent demographic trends in the number of cases and management of AMI through the first 12 months of the COVID-19 pandemic. Methods and Results: We assessed demographic trends, patient characteristics, and AMI outcomes (n=730) during the first 12 months of the COVID-19 pandemic and compared them with corresponding months during the control period (February 2016-January 2020; n=2,742) using data from the Mie ACS Registry. Although a 25.8% reduction in hospitalizations for AMI was observed in the 3 months following the declaration of a state of emergency (47.7 vs. 64.3/month; P=0.002), the total number of AMI patients was similar between the 12-month COVID-19 and control periods (60.8 vs. 57.2/month; P=0.58). The number of patients requiring direct ambulance transport was lower in the first half of the COVID-19 than control period (44.4% vs. 51.5; P=0.028). In-hospital mortality was higher in the second half of the COVID-19 than control period (8.9% vs. 5.8%; P=0.032). Conclusions: Through the first 12 months of the COVID-19 pandemic, the number of AMI cases was similar to that in previous years. The COVID-19 pandemic changed the behavior of AMI patients and both pre- and in-hospital medical management, which significantly affected the severity and prognosis of AMI.

Prognostic Impact of Peak Aortic Jet Velocity on Patients With Acute Myocardial Infarction.

BACKGROUND: Aortic valve stenosis (AS) leads to increased cardiovascular mortality and morbidity, and recent studies reported that even mild-to-moderate AS was associated with poor prognosis in the general population. This study investigated the prognostic impact of mild or moderate AS, defined as 2.0 m/s ≤ peak aortic jet velocity (Vmax) ≤3.9 m/s using echocardiography in acute myocardial infarction (AMI) patients.Methods and Results: This study enrolled 3,049 AMI patients using data from the Mie ACS registry. Patients were divided into 2 groups according to Vmax: Group 1: Vmax <2.0 m/s and/or visually intact aortic valve in which all 3 leaflets are fully and evenly open; Group 2: 2.0 m/s ≤ Vmax ≤ 3.9 m/s. There were 2,976 patients in Group 1and 73 patients in Group 2. The Group 2 patients were older, had a higher percentage of males and had lower body mass index and Killip ≥2 than the Group 1 patients. Angiographic data, door-to-balloon time, and mechanical support were not different between the 2 groups. The Group 2 patients demonstrated a significantly higher all-cause mortality rate (P<0.01) and composite of cardiovascular death and heart failure hospitalization (P<0.01), and Kaplan-Meier analysis showed the same tendency in propensity score-matched patients. CONCLUSIONS: The present study revealed that mild or moderate AS based on Vmax is associated with poor prognosis following AMI.

Perinatal hypoxia aggravates occlusive pulmonary vasculopathy in SU5416/hypoxia-treated rats later in life.

Pulmonary arterial hypertension (PAH) is a fatal disease, which is characterized by occlusive pulmonary vascular disease (PVD) in small pulmonary arteries. It remains unknown whether perinatal insults aggravate occlusive PVD later in life. We tested the hypothesis that perinatal hypoxia aggravates PVD and survival in rats. PVD was induced in rats with/without perinatal hypoxia (embryonic day 14 to postnatal day 3) by injecting SU5416 at 7 wk of age and subsequent exposure to hypoxia for 3 wk (SU5416/hypoxia). Hemodynamic and morphological analyses were performed in rats with/without perinatal hypoxia at 7 wk of age (baseline rats, n = 12) and at 15 wk of age in 4 groups of rats: SU5416/hypoxia or control rats with/without perinatal hypoxia (n = 40). Pulmonary artery smooth muscle cells (PASMCs) from the baseline rats with/without perinatal hypoxia were used to assess cell proliferation, inflammation, and genomic DNA methylation profile. Although perinatal hypoxia alone did not affect survival, physiological, or pathological parameters at baseline or at the end of the experimental period in controls, perinatal hypoxia decreased weight gain and survival rate and increased right ventricular systolic pressure, right ventricular hypertrophy, and indices of PVD in SU5416/hypoxia rats. Perinatal hypoxia alone accelerated the proliferation and inflammation of cultured PASMCs from baseline rats, which was associated with DNA methylation. In conclusion, we established the first fatal animal model of PAH with worsening hemodynamics and occlusive PVD elicited by perinatal hypoxia, which was associated with hyperproliferative, proinflammatory, and epigenetic changes in cultured PASMCs. These findings provide insights into the treatment and prevention of occlusive PVD.

CRISPR-mediated Bmpr2 point mutation exacerbates late pulmonary vasculopathy and reduces survival in rats with experimental pulmonary hypertension.

BACKGROUND: Patients with pulmonary arterial hypertension (PAH) carrying bone morphogenetic protein receptor type 2 (Bmpr2) mutations present earlier with severe hemodynamic compromise and have poorer survival outcomes than those without mutation. The mechanism underlying the worsening clinical phenotype of PAH with Bmpr2 mutations has been largely unaddressed in rat models of pulmonary hypertension (PH) because of the difficulty in reproducing progressive PH in mice and genetic modification in rats. We tested whether a clinically-relevant Bmpr2 mutation affects the progressive features of monocrotaline (MCT) induced-PH in rats. METHODS: A monoallelic single nucleotide insertion in exon 1 of Bmpr2 (+/44insG) was generated in rats using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9, then PH, pulmonary vascular disease (PVD) and survival after MCT injection with or without a phosphodiesterase type 5 inhibitor, tadalafil, administration were assessed. RESULTS: The +/44insG rats had reduced BMPR2 signalling in the lungs compared with wild-type. PH and PVD assessed at 3-weeks after MCT injection were similar in wild-type and +/44insG rats. However, survival at 4-weeks after MCT injection was significantly reduced in +/44insG rats. Among the rats surviving at 4-weeks after MCT administration, +/44insG rats had increased weight ratio of right ventricle to left ventricle plus septum (RV/[LV + S]) and % medial wall thickness (MWT) in pulmonary arteries (PAs). Immunohistochemical analysis showed increased vessels with Ki67-positive cells in the lungs, decreased mature and increased immature smooth muscle cell phenotype markers in the PAs in +/44insG rats compared with wild-type at 3-weeks after MCT injection. Contraction of PA in response to prostaglandin-F2α and endothelin-1 were significantly reduced in the +/44insG rats. The +/44insG rats that had received tadalafil had a worse survival with a significant increase in RV/(LV + S), %MWT in distal PAs and RV myocardial fibrosis compared with wild-type. CONCLUSIONS: The present study demonstrates that the Bmpr2 mutation promotes dedifferentiation of PA smooth muscle cells, late PVD and RV myocardial fibrosis and adversely impacts both the natural and post-treatment courses of MCT-PH in rats with significant effects only in the late stages and warrants preclinical studies using this new genetic model to optimize treatment outcomes of heritable PAH.

Cardiorenal protective effects of sodium-glucose cotransporter 2 inhibition in combination with angiotensin II type 1 receptor blockade in salt-sensitive Dahl rats.

OBJECTIVE: The kidney plays a central role in regulating the salt sensitivity of blood pressure (BP) by governing sodium excretion and reabsorption via renal sodium transporters. We hypothesized that sodium-glucose cotransporter 2 (SGLT2) inhibition and angiotensin II type 1 receptor (AT1R) blockade can synergistically reduce renal sodium reabsorption by beneficially effects on these transporters, leading to lower BP and ameliorating renal and cardiac damage. METHODS AND RESULTS: Dahl salt-sensitive rats were treated orally for 8weeks with a normal salt diet (0.3% NaCl), a high-salt diet (8% NaCl), high-salt diet with ipragliflozin (0.04%), high-salt diet with losartan (0.05%) or high-salt diet with a combination of ipragliflozin and losartan. The combination treatment significantly reduced BP and increased daily urine sodium excretion compared with losartan or ipragliflozin monotherapy, leading to greater improvement in BP salt sensitivity than ipragliflozin monotherapy. The combination treatment significantly ameliorated glomerulosclerosis and reduced cardiomyocyte hypertrophy compared with losartan or ipragliflozin monotherapy. The protein expression levels of Na+/H+ exchanger isoform 3 (NHE3) and Na+-K+-CI- cotransporter 2 (NKCC2) in the kidney were significantly decreased with losartan monotherapy and combination treatment, but not with ipragliflozin monotherapy. CONCLUSION: Inhibition of SGLT2 in combination with an angiotensin II receptor blocker effectively improved BP salt sensitivity by reducing renal expression levels of sodium transporters including NHE3 and NKCC2, which eventually led to improvement of BP salt sensitivity and cardiorenal protection.

Serum-Induced Expression of Brain Natriuretic Peptide Contributes to Its Increase in Patients with HFpEF.

Brain natriuretic peptide (BNP) levels are increased in both patients with heart failure with preserved (HFpEF) and reduced ejection fraction (HFrEF), but the reasons for this remain unclear. Our purpose was to examine whether serum-induced BNP (iBNP) expression partly contributes to increased BNP in patients with HFpEF. BNP reporter cardiomyocytes from pBNP-luc-KI mice were stimulated with serum from patients with HFpEF or HFrEF (n = 114 and n = 82, respectively). Luciferase activity was examined as iBNP and the iBNP-to-BNP ratio was evaluated. Patient characteristics and clinical parameters were compared, and multivariate regression analysis was performed to determine independent predictors of the iBNP-to-BNP ratio. Female sex and frequencies of atrial fibrillation, hypertension and the use of a calcium channel blocker (CCB) were higher in HFpEF. The iBNP-to-BNP ratio was significantly higher in HFpEF (26.9) than in HFrEF (16.1, p < 0.001). Multivariate regression analysis identified the existence of HFpEF as an independent predictor of the iBNP-to-BNP ratio after adjusting for all other measurements (ß = 0.154, p = 0.032). Age, hemoglobin, CCB usage and deceleration time were also independent predictors (ß = 0.167, p = 0.025; ß = 0.203, p = 0.006; ß = 0.138, p = 0.049; and ß = 0.143, p = 0.049, respectively). These results indicate that the elevated BNP in patients with HFpEF is partly due to iBNP from the heart.

Regulation of myosin light-chain phosphorylation and its roles in cardiovascular physiology and pathophysiology.

The regulation of muscle contraction is a critical function in the cardiovascular system, and abnormalities may be life-threatening or cause illness. The common basic mechanism in muscle contraction is the interaction between the protein filaments myosin and actin. Although this interaction is primarily regulated by intracellular Ca2+, the primary targets and intracellular signaling pathways differ in vascular smooth muscle and cardiac muscle. Phosphorylation of the myosin regulatory light chain (RLC) is a primary molecular switch for smooth muscle contraction. The equilibrium between phosphorylated and unphosphorylated RLC is dynamically achieved through two enzymes, myosin light chain kinase, a Ca2+-dependent enzyme, and myosin phosphatase, which modifies the Ca2+ sensitivity of contractions. In cardiac muscle, the primary target protein for Ca2+ is troponin C on thin filaments; however, RLC phosphorylation also plays a modulatory role in contraction. This review summarizes recent advances in our understanding of the regulation, physiological function, and pathophysiological involvement of RLC phosphorylation in smooth and cardiac muscles.

Assessment of Pseudocoarctation of the Aorta with Saccular Aneurysms by Four-Dimensional Flow Magnetic Resonance Imaging and Histological Analysis.

In this study, we present the case of a 21-year-old woman with pseudocoarctation of the aorta with saccular aneurysms that were evaluated by four-dimensional flow magnetic resonance imaging and histological analysis. We observed complete occupation of the aneurysm sacs by vortex flow and high peak wall shear stress in the proximal region of the kinked aorta. The aortic replacement was performed for the thoracic aortic aneurysms and the clinical course was uneventful. The aneurysms were histopathologically diagnosed as pseudoaneurysms based on the disappearance of all three layers and their replacement with collagen-rich connective tissues. These findings indicate that abnormal flow dynamics and the resulting abnormal shear stress in the aorta may play central roles in the formation and development of a saccular aneurysm.

Effect of left ventricular ejection fraction on the prognostic impact of chronic total occlusion in a non-infarct-related artery in patients with acute myocardial infarction.

BACKGROUND: Chronic total occlusion (CTO) in a non-infarct-related artery (IRA) in patients with acute coronary syndrome (ACS) is associated with a poor prognosis. However, whether the prognostic impact of non-IRA CTO differs according to left ventricular ejection fraction (LVEF) is unclear. METHODS AND RESULTS: A total of 2060 consecutive acute myocardial infarction (AMI) patients who underwent primary percutaneous coronary intervention (PCI) were classified into 2 groups according to their LVEF (reduced EF: LVEF < 50%, preserved EF: LVEF ≥ 50%) and further subdivided according to the presence of concomitant non-IRA CTO. In the reduced EF group, patients with CTO had a higher 1-year all-cause death rate (20.3% vs. 34.3%, P = 0.001) and major adverse cardiac event rate (MACE: 19.6% vs. 39.6%, P < 0.001) compared to those without CTO, but they were similar between patients with and without CTO in the preserved EF group. Non-IRA CTO was an independent predictor of all-cause death (HR 1.58, 95% CI 1.06-2.33, P = 0.02) and MACE (HR 1.67, 95% CI 1.14-2.46, P = 0.009) only in the reduced EF group. In addition, the outcomes of successful CTO-PCI seemed to be similar to those without CTO in the reduced EF group. CONCLUSIONS: CTO in a non-IRA may contribute to a poor prognosis only in AMI patients with reduced LVEF.

Semiconductor property imaging on as-grown wafer with monochromatic tunable THz-wave source.

In this paper, we propose the use of a tunable monochromatic terahertz (THz) wave source to measure the carrier properties of semiconductors. We also report on the recent improvement of our measurement system and demonstrate its ability to measure an as-grown sample; our system involves reference mirror-free reflective measurement. In our method, carrier properties, such as resistivity and mobility in semiconductors relating to the carrier density, can be determined by reflective measurement. This method was applied to composite semiconductors-GaN, GaAs, and SiC. In the measurement system, we improved the filter to block the infrared beam co-linearly propagating with the desired THz-wave. Consequently, the signal intensity was 13 times higher than that achieved in our previous work. We demonstrated the measured carrier properties in an as-grown n-type GaN wafer. We observed flat features via our THz-reflective measurement, whereas the sample showed prominent surface roughness in a picture taken with visible light.

Quadratic nonlinear optical properties of the organic N-benzyl-2-methyl-4-nitroaniline (BNA) biaxial crystal.

We performed the direct measurement of second harmonic generation and sum frequency generation phase-matching directions in the organic N-benzyl-2-methyl-4-nitroaniline crystal over its visible and near-infrared transparency range. The fit of these data allowed us to refine the Sellmeier equations of the three principal refractive indices in this range. With these equations, we improved the calculated tuning curves of terahertz emission from a phase-matched difference frequency process. We also determined the absolute magnitude of the d24 nonlinear coefficient.

[Molecular Investigation on the Presence of Hepatitis E Virus in Wild Game in Okazaki City, Japan between April 2010 and November 2014].

Cases of hepatitis E have been caused by infection with hepatitis E virus (HEV) due to consumption of raw or undercooked game meats and liver of wild boars or deer in various countries. We investigated HEV prevalence in wild boar and deer in Okazaki City and its outskirts, Aichi, Japan, using liver samples (439 boar; 185 deer) collected between April 2010 and November 2014. HEV RNA was detected in 49 (11.2%)of 439 and 0 of 185 samples of wild boar and deer, respectively. The positive rate (13.0%, 28/216) of HEV RNA from the wild boar with estimated body weight of less than 40 kg was significantly higher than that (2.7%, 3/111) in animals with estimated body weight of more than 40 kg. The 49 HEV strains were typed as genotype 4 (G4) by phylogenetic analysis. They clustered with Aichi/Shizuoka strains and 48 of the 49 strains subclustered together (Okazaki strains).

Crossover from capacitive to inductive electromagnetic responses in near self-complementary metallic checkerboard patterns.

Transmission spectra of near-self-complementary metallic checkerboard patterns (MCPs) exhibit a drastic change when the metal squares are brought into contact with each other from a noncontact state. Transmission spectra of near-self-complementary samples, which are fabricated by printing technology, show rather gradual systematic change with changing the nominal metal square size while keeping the period because of randomness naturally introduced by the limited accuracy of the printer. The spectra have transmission-invariant frequencies, which means that the spectra are a superposition of two types of spectra, the ratio of which depends on the nominal square size. The correlation seen in the real and imaginary parts of the complex amplitude spectra can be interpreted based on the Kramers-Kronig relation. As an application of the sensitiveness of the transmission spectrum of the MCPs to connectivity of the metal squares, the revealing of an optically hidden pattern by a terahertz beam is demonstrated.

Tunable Terahertz-wave generation from DAST crystal pumped by a monolithic dual-wavelength fiber laser.

For developing a continuous-wave (CW) tunable Terahertz-wave (THz-wave) source using difference-frequency generation (DFG) in highly nonlinear optical crystals, we proposed and demonstrated a dual-wavelength fiber ring laser system operating around 1060 nm based on wideband chirped fiber Bragg gratings (CFBGs) and semiconductor optical amplifier (SOA). Thermo-induced phase shift along the CFBG produces a very sharp transmission spike therefore two lasing wavelengths with single longitudinal mode operation are oscillating simultaneously within the fiber ring cavity. Due to the inhomogeneous gain broadening property of SOA, the wavelength spacing of our dual-wavelength fiber laser can be continuously adjusted from 0.3 to 9.5 nm. By using this single emitter dual-wavelength fiber laser to pump an organic nonlinear DAST crystal, type-0 collinear phase matching of DFG process can be fulfilled and monochromatic THz wave ranging from 0.5 to 2 THz has been successfully generated.

Composite THz materials using aligned metallic and semiconductor microwires, experiments and interpretation.

We report fabrication method and THz characterization of composite films containing either aligned metallic (tin alloy) microwires or chalcogenide As2Se3 microwires. The microwire arrays are made by stack-and-draw fiber fabrication technique using multi-step co-drawing of low-melting-temperature metals or semiconductor glasses together with polymers. Fibers are then stacked together and pressed into composite films. Transmission through metamaterial films is studied in the whole THz range (0.1-20 THz) using a combination of FTIR and TDS. Metal containing metamaterials are found to have strong polarizing properties, while semiconductor containing materials are polarization independent and could have a designable high refractive index. Using the transfer matrix theory, we show how to retrieve the complex polarization dependent refractive index of the composite films. Finally, we study challenges in the fabrication of metamaterials with sub-micrometer metallic wires by repeated stack-and-draw process by comparing samples made using 2, 3 and 4 consecutive drawings. When using metallic alloys we observe phase separation effects and nano-grids formation on small metallic wires.

Study of water concentration measurement in thin tissues with terahertz-wave parametric source.

Water concentration and distribution in biotissues are important factors in many applications. THz-wave is a viable tool for water content measurement due to its highly sensitivity to water. In this study, the measuring errors of water concentration using THz-wave induced by transmittance and sample thickness were analyzed theoretically. The chosen basis for sample thickness and measuring THz frequency were presented theoretically. Measurements of the water two-dimensional mapping in different animal tissue samples were demonstrated experimentally, which clearly shows the spatial distribution of the tissues.