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Background: Fulminant myocarditis presentation (FMP) is a rare and severe presentation of myocarditis. The natural history of FMP and its clinical features associated with poor outcomes are incompletely understood because there is a lack of generalizable evidence. Methods: This multicenter retrospective cohort study included patients hospitalized with histologically proven myocarditis who underwent catecholamine or mechanical support from 235 cardiovascular training hospitals across Japan between April 2012 and March 2017. Clinical features and the prognostic predictors of death or heart transplantation within 90 days on the basis of clinical and pathologic findings were determined using the Kaplan-Meier method, log-rank test, and Cox regression analysis. Results: This study included 344 patients with histologically proven FMP (median age, 54 years; 40% female). The median follow-up was 600 days (interquartile range, 36 to 1599 days) and the cumulative risk of death or heart transplantation at 90 days was 29% (n=98). Results from multivariable Cox regression analysis showed that older age, nonsinus rhythm, low left ventricular wall motion ( Conclusions: The results from analyses of data from this multicenter registry demonstrated that patients with FMP are at a higher risk of death or heart transplantation in real-world settings. These observations inform which clinical and pathologic findings may be useful for prognostication in FMP. Registration: URL: https://www.umin.ac.jp/ctr ; Unique identifier: UMIN000039763.

In recent years, transthyretin amyloid cardiomyopathy (ATTR-CM) has received increasing attention; however, the epidemiology of ATTR-CM in Japan is not yet understood. In the Kumamoto Cardiac Amyloid Survey, we evaluated the current incidence, clinical characteristics, diagnostic approaches, and treatment strategies for ATTR-CM and compared tafamidis-prescription hospitals with regional hospitals.We conducted a retrospective multicenter observational cohort study. The registry included patients with ATTR-CM diagnosed in two tafamidis-prescription hospital institutes [Japanese Circulation Society (JCS)-certified facilities] and 15 regional cardiovascular facilities in Kumamoto between January 2018 and December 2020.In total, 174 patients were diagnosed with ATTR-CM. The incidence of ATTR-CM was estimated to be approximately 1 per 10,000 person-years in the elderly population (65 years old) in Kumamoto. Compared with that in the JCS-certified facilities cohort (n=115), age at diagnosis was significantly older (84.5 ± 5.6 vs. 77.5 ± 6.3 years old; p0.01) in the regional hospitals cohort (n=59). Histological (25% vs. 81%; p0.01) and genetic diagnosis (7% vs. 82%) were also less frequently performed. Probable (as indicated by positive bone scintigraphy findings with confirmation of monoclonal protein absence) and possible (as indicated by positive bone scintigraphy findings without confirmation of monoclonal protein absence) ATTR-CM accounted for the majority of cases (75% vs. 19%; p0.01) in the regional hospitals cohort compared to the JCS-certified facilities cohort. There were no cases of hereditary ATTR-CM among the patients who underwent TTR genetic testing (n=98).We confirmed the incidence of ATTR-CM in Kumamoto and the diagnostic approach used in patients with ATTR-CM. Further prospective studies with a larger sample are needed to validate our results and to further shed light on the epidemiology of ATTR-CM in Japan.

This study uses a Japanese nationwide sample and experimentally compares rebate and matching, both of which are schemes intended to lower the price of monetary donation. Standard economic theory predicts that the two schemes will have the same effect on individuals’ donation behavior when their donation price is equivalent. However, we conduct an incentivized economic experiment through the Internet on 2300 Japanese residents, and find that matching, which lowers the donation price by adding a contribution from a third-party, increases individuals’ donation expenditures compared to rebate, which lowers it through a refund from a third-party. The experimental result shows that the donation expenditure in a 50% rebate treatment drops by approximately 126 Japanese yen compared to the control, while in a 1:1 matching treatment with essentially the same price of donation as the 50% rebate, the expenditure conversely rises by approximately 56 Japanese yen. This tendency is consistent with the results of previous experimental studies comparing the two schemes. We further empirically confirm that the superiority of 1:1 matching over 50% rebate is not conclusively influenced by the participants’ confusion or misunderstanding, or budget constraint lines’ difference between the two schemes. Although the Japanese government has previously enriched rebate’s content, the level of monetary donations by the Japanese people is still low on an international scale. Based on this study’s findings, we discuss the possibility that implementing matching into the society effectively encourages their donation behavior.

This study conducted randomized controlled trials to evaluate the effectiveness of an environmental education class and the impacts of nudges and boosts implemented in this program on high school students' basic knowledge, attitudes, and behaviors regarding environmental issues in Japan. This environmental education class consisted of a lecture on reducing the use of plastic products for energy conservation in daily life, a board game for learning how to reduce plastic waste, and a worksheet for reflection. Four types of worksheets were randomly distributed: nudges, in which students were asked to set a goal regarding their level of effort in not throwing away plastic products such as plastic bags, wet wipes, and plastic bottles; boosts, in which participants were asked to write an essay to help increase their empathy for the parties impacted by environmental issues; both nudges and boosts; and none (neither nudges nor boosts). After environmental education, an end-line survey was conducted to evaluate the effectiveness of this class. This study found that the environmental education class significantly improved students' basic environmental knowledge and promoted their concerns about plastic waste. Although there was no evidence that nudges and boosts amplify the effects of the environmental education class on the students' knowledge, nudges were successful in making them more concerned about plastic waste. The results showed that students who received nudges or boosts were more likely to refuse free wet wipes offered at convenience stores but were not more likely to refuse plastic bottles. These results also indicated that interventions through environmental education can change students' pro-environmental behaviors only if the cost of behavioral change is low. The environmental education class not only increased students' environmental knowledge and attitudes, but the use of worksheets in administering nudges and boosts ensured the effectiveness of environmental education.

There have been numerous studies in which the biological role of oxytocin in trusting behavior has been investigated. However, a link between oxytocin and trust in humans was discovered only in one early study. We hypothesized that there is a large interindividual variation in oxytocin sensitivity, and that such variation is one reason for the doubt surrounding the role of oxytocin in trusting behavior. Here, in a double-blind, prospective, case-control study, we administered intranasal oxytocin to participants of trust and risk games. We measured salivary oxytocin concentration, relating it to the amount of money transferred among participants (a proxy for trust) and the autism-spectrum quotient (AQ). A one-sided Fisher’s exact test was performed to detect differences between the oxytocin and placebo groups in the proportions of investors who transferred the maximum amount of money. We discovered a tendency for participants who received oxytocin to transfer higher amounts of money to co-participants than those who received a placebo (P = 0.04). We also revealed a high degree of interindividual variation in salivary oxytocin concentrations after oxytocin administration. After stratifying the samples with respect to oxytocin sensitivity, oxytocin-sensitive participants in the oxytocin group also transferred higher amounts of money than those in the placebo group (P = 0.03), while such a tendency was not observed for oxytocin-insensitive participants (P = 0.34). Participants with lower AQ scores (less severe autistic traits) exhibited a greater tendency toward trusting behavior after oxytocin administration than did those with higher AQ scores (P = 0.02). A two-sample t-test that was performed to detect significant differences in the mean transfers between the oxytocin and placebo groups indicated no significant between-group difference in the mean transfers (P = 0.08). There are two possible interpretations of these results: First, there is no effect of oxytocin on trust in humans; second, the effects of oxytocin on trust in humans is person-dependent. However, the results should be interpreted with caution as the effect size was not larger than the minimal detectable effect size and the results were not statistically significant (P > 0.05) after Bonferroni corrections.

Nudge-based messages have been employed in various countries to encourage voluntary contact-avoidance and infection-prevention behaviors to control the spread of COVID-19. People have been repeatedly exposed to such messages; however, whether the messages keep exerting a significant impact over time remains unclear. From April to August 2020, we conducted a four-wave online survey experiment to examine how five types of nudge-based messages influence Japanese people’s self-reported preventive behaviors. In particular, we investigate how their behaviors are affected by repeated displays over time. The analysis with 4241 participants finds that only a gain-framed altruistic message, emphasizing their behavioral adherence would protect the lives of people close to them, reduces their frequency of going out and contacting others. We do not find similar behavioral changes in messages that contain an altruistic element but emphasize it in a loss-frame or describe their behavioral adherence as protecting both one’s own and others’ lives. Furthermore, the behavioral change effect of the gain-framed altruistic message disappears in the third and fourth waves, although its impact of reinforcing intentions remains. This message has even an adverse effect of worsening the compliance level of infection-prevention behaviors for the subgroup who went out less frequently before the experiment. The study’s results imply that when using nudge-based messages as a countermeasure for COVID-19, policymakers and practitioners need to carefully scrutinize the message elements and wording and examine to whom and how the messages should be delivered while considering their potential adverse and side effects.

This study uses a Japanese nationwide sample and experimentally compares rebate and matching, both of which are schemes intended to lower the price of monetary donation. Standard economic theory predicts that the two schemes will have the same effect on individuals’ donation behavior when their donation price is equivalent. However, we conduct an incentivized economic experiment through the Internet on 2,300 Japanese residents, and find that matching, which lowers the donation price by adding a contribution from a third-party, increases individuals’ donation expenditures compared to rebate, which lowers it through a refund from a third-party. Specifically, the experimental result shows that the donation expenditure in a 50% rebate treatment drops by approximately ¥126 compared to the control, while in a 1:1 matching treatment with essentially the same price of donation as the 50% rebate, the expenditure conversely rises by approximately ¥56. This tendency is consistent with the results of previous experimental studies comparing the two schemes. We further empirically confirm that the superiority of 1:1 matching over 50% rebate is not conclusively influenced by the participants’ confusion or misunderstanding, or budget constraint lines’ difference between the two schemes. Although the Japanese government have previously enriched rebate’s content, the level of monetary donations by the Japanese people is still low on an international scale. Based on this study’s findings, we discuss the possibility that implementing matching into the society effectively encourages their donation behavior.

cis-Prenyltransferases (cis-PTs) catalyze consecutive condensations of isopentenyl diphosphate to an allylic diphosphate acceptor to produce a linear polyprenyl diphosphate of designated length. Dimer formation is a prerequisite for cis-PTs to catalyze all cis-prenyl condensation reactions. The structure-function relationship of a conserved C-terminal RXG motif in cis-PTs that forms inter-subunit interactions and has a role in catalytic activity has attracted much attention. Here, we solved the crystal structure of a medium-chain cis-PT from Thermobifida fusca that produces dodecaprenyl diphosphate as a polyprenoid glycan carrier for cell wall synthesis. The structure revealed a characteristic dimeric architecture of cis-PTs in which a rigidified RXG motif of one monomer formed inter-subunit hydrogen bonds with the catalytic site of the other monomer, while the RXG motif of the latter remained flexible. Careful analyses suggested the existence of a possible long-range negative cooperativity between the two catalytic sites on the two monomeric subunits that allowed the binding of one subunit to stabilize the formation of the enzyme-substrate ternary complex and facilitated the release of Mg-PPi and subsequent intra-molecular translocation at the counter subunit so that the condensation reaction could occur in consecutive cycles. The current structure reveals the dynamic nature of the RXG motif and provides a rationale for pursuing further investigations to elucidate the inter-subunit cooperativity of cis-PTs.

Oxytocin-enhanced prosocial behaviour depends on individual characteristics. This study investigated the relationship between oxytocin and competitiveness, which is another important social trait and predicts economic and social outcomes. In this double-blind, randomized, and placebo-controlled study of 192 male participants, we examined whether oxytocin moderates competitiveness and whether the effect of oxytocin on competitiveness is amplified in individuals with autistic traits. While our results show no relationship between oxytocin and competitiveness, we observed suggestive patterns: albeit not significantly, oxytocin reduced and enhanced competitiveness among participants without autistic traits and among their counterparts with autistic traits, respectively.

Objectives: We aimed to investigate the usefulness of inserting a 6Fr sheath guided by duplex ultrasonography via a popliteal artery puncture. We also aimed to demonstrate endoluminal tracking using a retrograde approach using the Gogo catheter with intravascular ultrasound (IVUS). Background: The bidirectional approach is useful for increasing the success rate of the procedure for long superficial femoral artery–chronic total occlusions (SFA-CTOs). However, this procedure becomes somewhat complicated. Since the proximal blood vessel diameter is clearly larger than the distal end of the CTO and the body surface duplex guide can also be used in the proximal part, it is easier to introduce a retrograde guidewire (GW) into the proximal end. Methods: We performed endovascular treatment for long SFA-CTOs with a Gogo catheter + IVUS guide in 31 consecutive cases (male 20/female 11; mean age, 75.6 ± 7.6) from May 2017 to November 2018. We advanced the IVUS until the true lumen could be confirmed and advanced the Gogo catheter toward the IVUS for reinforcement. We attempted to approach the long CTO by repeating this procedure. We named this procedure the GIP method (GIP: Gogo catheter with IVUS via a popliteal puncture). Hemostasis of the popliteal artery was achieved using a commercially available compression hemostatic kit (Tometa-kun, XEMEX, Japan). Results: Successful revascularization was achieved in all cases (in 2 cases, a femoral artery puncture was added, and a bidirectional approach was used, and in 1 case, a CROSSER system was used). On average, the fluoroscopy time was 42.2 ± 30.4 minutes, radiation dose 93.7 ± 78.7 mGy, and amount of contrast medium used 15.0 ± 9.6 mL. The procedure time was defined as from the start of the popliteal artery puncture to the time the GW passed through the CTO lesion, including the posture transformation time from prone to the supine position. The procedure time was 42.1 ± 40.2 minutes. There were no major adverse events or other major complications, such as a distal embolism, rupture of the CTO lesion, arteriovenous fistula, or major hematoma requiring a transfusion or surgical treatment. Only 2 small hematomas occurred at the popliteal artery puncture site. The patients were treated conservatively and were discharged as usual. Conclusions: Endovascular treatment of long SFA-CTOs via the popliteal approach was effective and safe. Using the GIP method to address long SFA-CTOs is recommended.

We investigate the labor supply reaction to wage cuts and tax increases using a real-effort experiment. First, subjects perform a task and receive a reward with taxes deducted. Second, the wage cut treatment reduces the wage rate and the tax increase treatment increases the tax; wage and tax remain unchanged for the control group. The real wage is equal for both treatments, and both treatments had significantly smaller increases in effort levels than the control group. The effort level change did not differ between the treatments. Therefore, the net wage illusion and tax aversion identified in previous studies are absent.

Case 1. A 45-year-old man with dyspnoea. In his echocardiography, we detected a 35×15 mm left ventricular thrombus (LVT) with a low ejection fraction. Conventional therapy (unfractionated heparin combined with vitamin K antagonists) for the LVT resulted in an acute inferior myocardial infarction

A 78-year-old man presented with effort angina pectoris. Coronary angiography revealed a diffuse 75%–90% stenosis with calcification in the mid-left anterior descending artery. In advanced stenotic lesions with calcifications, coronary stents can be stacked even after frequent predilatations. It

We empirically investigated how voluntary helping behavior is influenced by the number of its potential recipients by using a nationwide survey in Japan (N = 1,333) and examining the relationship between blood type and blood donation behavior. It is generally known in Japan that type O blood can be medically transfused to individuals of all blood groups; therefore, the potential transfusion recipients’ number of the type O blood is the largest among the four blood groups. Our empirical analysis revealed that people with type O blood were more likely to have donated blood in the past than those with the other blood types. This association was stronger in a subsample of individuals who had knowledge relating to the above-mentioned widespread utility of type O blood. In addition, our empirical analysis arrested the concern that potential blood-type differences in altruistic attitudes might explain their differences in blood donation behavior, by confirming that type O blood was not significantly related to other altruistic behaviors (i.e., registration for bone-marrow donation, intention to donate organs, and making monetary donations) or attitudes (i.e., general trust, altruism, reciprocity, and agreeableness). After conducting further analyses, we concluded that the vast number of potential recipients of type O blood causes different patterns of Japanese people’s blood donation behavior across the four blood groups. This study added to the literature the real-world evidence concerning how having many potential recipients affect people’s behavior of providing common goods..

Objective Mitochondrial dysfunction plays an important role in cellular senescence and impaired function of vascular endothelium, resulted in cardiovascular diseases. Telmisartan is a unique angiotensin II type I receptor blocker that has been shown to prevent cardiovascular events in high risk patients. AMP-activated protein kinase (AMPK) plays a critical role in mitochondrial biogenesis and endothelial function. This study assessed whether telmisartan enhances mitochondrial function and alters cellular functions via AMPK in human coronary artery endothelial cells (HCAECs). Methods and results In cultured HCAECs, telmisartan significantly enhanced mitochondrial activity assessed by mitochondrial reductase activity and intracellular ATP production and increased the expression of mitochondria related genes. Telmisartan prevented cellular senescence and exhibited the anti-apoptotic and pro-angiogenic properties. The expression of genes related anti-oxidant and pro-angiogenic properties were increased by telmisartan. Telmisartan increased endothelial NO synthase and AMPK phosphorylation. Peroxisome proliferator-activated receptor gamma signaling was not involved in telmisartan-induced improvement of mitochondrial function. All of these effects were abolished by inhibition of AMPK. Conclusions Telmisartan enhanced mitochondrial activity and exhibited anti-senescence effects and improving endothelial function through AMPK in HCAECs. Telmisartan could provide beneficial effects on vascular diseases via enhancement of mitochondrial activity and modulating endothelial function through AMPK activation.

A 69-year-old man presented to our hospital with chest pain. Two-dimensional transthoracic echocardiography showed hypertrophy of the left ventricle, mid-ventricular obstruction and an apical aneurysm. Color-flow imaging at the obstruction site on the apical four-chamber view demonstrated systolic flow reversal in addition to a paradoxical jet flow. The systolic flow reversal may have been caused by a decreased apical contractility and pressure during systole.

BACKGROUND Several studies have reported that colchicine attenuated the infarct size and inflammation in acute myocardial infarction (MI). However, the sustained benefit of colchicine administration on survival and cardiac function after MI is unknown. It was hypothesized that the short-term treatment with colchicine could improve survival and cardiac function during the recovery phase of MI.Methods and Results:MI was induced in mice by permanent ligation of the left anterior descending coronary artery. Mice were then orally administered colchicine 0.1 mg/kg/day or vehicle from 1 h to day 7 after MI. Colchicine significantly improved survival rate (colchicine, n=48: 89.6% vs. vehicle, n=51: 70.6%, P

We show evidence that a rise in social status (e.g., receiving a prize) has both positive and negative causal effects on recipients’ longevity, by using a dataset that covers Japan's two most prestigious and traditional literary recognitions. Although the two prizes share several characteristics, there exists the key difference that one of them is awarded to yet unknown or emerging novelists while the candidates for the other include a larger number of already established novelists. The main results of Cox's proportional hazard model analysis reveal that (1) the recipients of the prize for yet unknown or emerging novelists exhibit lower mortality than unsuccessful nominees, and the increase in longevity is estimated at 1.7 years; (2) by contrast, the recipients of the prize mainly for more established novelists report higher mortality than unsuccessful nominees, and the decrease in longevity is estimated at 5.3 years. We discuss with additional empirical analysis that we are likely to find a life-prolonging effect from receiving a prize when candidates belong to a lower social stratum. In so doing, this study provides narrative explanations for why earlier studies show conflicting relationships between receiving a prize and recipients’ longevity.

Background— Dual antiplatelet therapy with aspirin and clopidogrel is widely used in patients with coronary stents. High residual platelet reactivity (high RPR) after dual antiplatelet therapy is associated with increased cardiovascular events. Endothelial function could affect platelet reactivity in vivo. We hypothesized that endothelial dysfunction could be associated with high RPR after dual antiplatelet therapy in patients with stable coronary artery disease. Methods and Results— We screened patients with stable coronary artery disease for cytochrome P450 (CYP) 2C19 genotypes and enrolled 103 patients who lacked CYP2C19*2 or *3 loss-of-function allele to minimize the effect of this gene on high RPR. All patients received aspirin (100 mg/d) and clopidogrel (75 mg/d for long-term treatment or a loading dose of 300 mg) before the following tests. Platelet aggregability was assessed as P2Y12 reaction unit using the VerifyNow System. High RPR was defined as P2Y12 reaction unit ≥230. Peripheral endothelial function was expressed as reactive hyperemia index using reactive hyperemia peripheral arterial tonometry. Fifty-three patients exhibited high RPR. High RPR patients were significantly older, had higher levels of B-type natriuretic peptide, and were predominantly hypertensive compared with non–high RPR patients. Reactive hyperemia index was significantly lower in high RPR patients (0.46±0.15) compared with non–high RPR patients (0.61±0.18; P r =−0.32; P =0.001). Multivariable logistic regression analysis identified reactive hyperemia index as an independent and significant determinant of high RPR (odds ratio, 0.55; 95% confidence interval, 0.39–0.78; P =0.001). Conclusions— In patients with stable coronary artery disease, endothelial function was significantly impaired in high RPR patients. Endothelial dysfunction is independently correlated with high RPR after dual antiplatelet therapy. Clinical Trial Registration— URL: http://www.umin.ac.jp/ctr . Unique identifier: UMIN000008239.

Background: Dipeptidyl peptidase 4 (DPP4) inhibitors are used for treatment of diabetes mellitus (DM). We hypothesized that sitagliptin, a DPP4-inhibitor, could improve endothelial dysfunction in DM patients with coronary artery disease (CAD). Methods and Results: The 40 patients with CAD and uncontrolled DM, aged 68.7±9.4 years (mean±standard deviation) (50% males, hemoglobin A1c [HbA1c] 7.4±1.0%) were assigned to either additional treatment with sitagliptin (50mg/day, n=20) or aggressive conventional treatment (control, n=20) for 6 months. Endothelial function was assessed by the reactive hyperemia peripheral arterial tonometry index (RHI). The clinical characteristics at baseline were not different between the groups. After treatment, fasting blood glucose and insulin levels, and lipid profiles were not different between the groups. HbA1c levels significantly improved similarly in both groups. The percent change in RHI was greater in the sitagliptin group than in the control group (62.4±59.2% vs. 15.9±22.0%, P

BACKGROUND Simple office-based counseling for diet and exercise does not appear to positively affect success rates in metabolic syndrome (MetS) patients. The utility of the lifestyle modification self-assessment score (Self-AS) in the improvement of endothelial function by office-based counseling for patients with MetS was investigated. METHODS AND RESULTS Patients with MetS (n=207) and age- and sex-matched individuals without MetS (n=124) were enrolled in this cross-sectional study. Endothelial function was assessed using reactive hyperemia-peripheral arterial tonometry index (RHI). Patients with MetS had significant endothelial dysfunction compared with those without MetS (RHI, 0.502±0.178 vs. 0.614±0.229; P

Background Worsening renal function, indicated by increased serum creatinine (SCr), is a common complication of percutaneous coronary procedures. Risk factors for increased SCr overlap with coronary risk factors involved in endothelial dysfunction. We hypothesized that endothelial dysfunction, measured using the reactive hyperemia peripheral arterial tonometry index (RHI), can predict periprocedure-increased SCr. Methods RHI was assessed before elective coronary procedures in 316 consecutive stable patients with a preserved estimated glomerular filtration rate (eGFR, >60 mL/min/1.73 m2). SCr was measured before and 2 days after procedures. Results There was no significant correlation between natural logarithmic transformations of RHI (Ln-RHI) and basal Ln-eGFR. Periprocedure increase in SCr was observed in 148 (47%) patients. The increased SCr group had significantly lower Ln-RHI [0.48 (0.36, 0.62) vs. 0.59 (0.49, 0.76), p

We show evidence that receiving Japan’s Akutagawa and Naoki Prizes for literature has positive and negative effects on their recipients’ longevity. Using a dataset covering both awards, we show that recipients of the Akutagawa Prize for rising novelists exhibit lower mortality than fellow nominees. The increase of longevity is estimated at 2.4 years. Recipients of the Naoki Prize for established novelists exhibit higher mortality than fellow nominees, and the decreased longevity is 5.1 years. These results indicate positive and negative causal effects from social status to longevity, and we identify and isolate those effects. In doing so, this study clarifies why earlier studies show conflicting relationships between receiving awards and the recipients’ longevity.

We test the equivalence of income and consumption taxes through a choice experiment. Under a given set of income and consumption parameters, subjects were asked to choose among an income tax of 20%, a consumption tax of 25% (which is an equivalent tax burden), a consumption tax of 22%, and a consumption tax of 20%. Our results showed that subjects prefer income tax to consumption tax when the nominal consumption tax rate is higher than the nominal income tax rate. However, subjects tend to prefer consumption tax to income tax when the nominal tax rates are identical. Our result, that subjects prefer income tax to consumption tax despite a higher tax burden, implies the consumption tax miscalculation bias. The consumption tax miscalculation bias is one where subjects miscalculate the amount of consumption tax as if it is declared by tax inclusive, as in the case of income tax, despite consumption tax being tax exclusive. If the income tax burden is equivalent to the consumption tax burden, subjects prefer income tax. This result implies that income and consumption taxes are not equivalent due to the consumption tax miscalculation bias.

ObjectivesThe aim of this study was to investigate the antiatherogenic effects of the dipeptidyl peptidase-4 inhibitor, des-fluoro-sitagliptin (DFS).BackgroundThe new class of anti–type 2 diabetes drugs, dipeptidyl peptidase-4 inhibitors, improves glucose metabolism by increasing levels of active glucagon-like peptide (GLP)-1.MethodsEndothelial function was examined by acetylcholine-induced endothelium-dependent vasorelaxation using aortic rings and atherosclerotic lesion development in the entire aorta in apolipoprotein E–deficient mice fed a high-fat diet with or without DFS, and the antiatherogenic effects of DFS were investigated in cultured human macrophages and endothelial cells. Plasma levels of active GLP-1 were measured in patients with or without coronary artery disease.ResultsDFS significantly improved endothelial dysfunction (89.9 ± 3.9% vs. 79.2 ± 4.3% relaxation at 10−4 mol/l acetylcholine, p < 0.05) associated with increased endothelial nitric oxide synthase phosphorylation and reduced atherosclerotic lesion area (17.7% [15.6% to 25.8%] vs. 24.6% [19.3% to 34.6%], p < 0.01) compared with vehicle treatment. In cultured human macrophages, DFS significantly increased GLP-1-induced cytosolic levels of cyclic adenosine monophosphate compared with GLP-1 alone, resulted in inhibiting phosphorylation of c-jun N-terminal kinase and extracellular signal-regulated kinase 1/2 and nuclear factor-kappa B p65 nuclear translocation through the cyclic adenosine monophosphate/protein kinase A pathway, and suppressed proinflammatory cytokines (i.e., interleukin-1-beta, interleukin-6, and tumor necrosis factor-alpha) and monocyte chemoattractant protein-1 production in response to lipopolysaccharide. DFS-enhanced GLP-1 activity sustained endothelial nitric oxide synthase phosphorylation and decreased endothelial senescence and apoptosis compared with GLP-1 alone. In the human study, fasting levels of active GLP-1 were significantly lower in patients with coronary artery disease than those without (3.10 pmol/l [2.40 to 3.62 pmol/l] vs. 4.00 pmol/l [3.10 to 5.90 pmol/l], p < 0.001).ConclusionsA DPP-4 inhibitor, DFS, exhibited antiatherogenic effects through augmenting GLP-1 activity in macrophages and endothelium.

Impaired selective turnover of p62 by autophagy causes severe liver injury accompanied by the formation of p62-positive inclusions and upregulation of detoxifying enzymes. These phenotypes correspond closely to the pathological conditions seen in human liver diseases, including alcoholic hepatitis and hepatocellular carcinoma. However, the molecular mechanisms and pathophysiological processes in these events are still unknown. Here we report the identification of a novel regulatory mechanism by p62 of the transcription factor Nrf2, whose target genes include antioxidant proteins and detoxification enzymes. p62 interacts with the Nrf2-binding site on Keap1, a component of Cullin-3-type ubiquitin ligase for Nrf2. Thus, an overproduction of p62 or a deficiency in autophagy competes with the interaction between Nrf2 and Keap1, resulting in stabilization of Nrf2 and transcriptional activation of Nrf2 target genes. Our findings indicate that the pathological process associated with p62 accumulation results in hyperactivation of Nrf2 and delineates unexpected roles of selective autophagy in controlling the transcription of cellular defence enzyme genes.

Keap1 is a substrate adaptor of a Cullin 3-based E3 ubiquitin ligase complex that recognizes Nrf2, and also acts as a cellular sensor for xenobiotics and oxidative stresses. Nrf2 is a transcriptional factor regulating the expression of cytoprotective enzyme genes in response to such stresses. Under unstressed conditions Keap1 binds Nrf2 and results in rapid degradation of Nrf2 through the proteasome pathway. In contrast, upon exposure to oxidative and electrophilic stress, reactive cysteine residues in intervening region (IVR) and Broad complex, Tramtrack, and Bric-à-Brac domains of Keap1 are modified by electrophiles. This modification prevents Nrf2 from rapid degradation and induces Nrf2 activity by repression of Keap1. Here we report the structure of mouse Keap1 homodimer by single particle electron microscopy. Three-dimensional reconstruction at 24-Å resolution revealed two large spheres attached by short linker arms to the sides of a small forked-stem structure, resembling a cherry-bob. Each sphere has a tunnel corresponding to the central hole of the β-propeller domain, as determined by x-ray crystallography. The IVR domain appears to surround the core of the β-propeller domain. The unexpected proximity of IVR to the β-propeller domain suggests that any distortions generated during modification of reactive cysteine residues in the IVR domain may send a derepression signal to the β-propeller domain and thereby stabilize Nrf2. This study thus provides a structural basis for the two-site binding and hinge-latch model of stress sensing by the Nrf2-Keap1 system.

Keap1, an adaptor protein of cullin-RING ubiquitin ligase complex, represses cytoprotective transcription factor Nrf2 in an oxidative stress-dependent manner. The accumulation of selective autophagy substrate p62 also activates Nrf2 target genes, but the detailed mechanism has not been elucidated. Crystal structure of Keap1-p62 complex revealed the structural basis for the Nrf2 activation in which Keap1 is inactivated by p62. The accumulation of p62 is observed in hepatocellular carcinoma. The activation of Nrf2 target genes, including detoxifying enzymes and efflux transporters, by p62 may protect the cancer cells from anti-cancer drugs.

cis-Prenyltransferases catalyze the consecutive condensation of isopentenyl diphosphate (IPP) with allylic prenyl diphosphates, producing Z,E-mixed prenyl diphosphate. The Mycobacterium tuberculosis Z,E-farnesyl diphosphate synthase Rv1086 catalyzes the condensation of one molecule of IPP with geranyl diphosphate to yield Z,E-farnesyl diphosphate and is classified as a short-chain cis-prenyltransferase. To elucidate the chain-length determination mechanism of the short-chain cis-prenyltransferase, we introduced some substitutive mutations at the characteristic amino acid residues of Rv1086. Among the mutants constructed, L84A showed a dramatic change of catalytic function to synthesize longer prenyl chain products than that of wild type, indicating that Leu84 of Rv1086 plays an important role in product chain-length determination. Mutagenesis at the corresponding residue of a medium-chain cis-prenyltransferase, Micrococcus luteus B-P 26 undecaprenyl diphosphate synthase also resulted in the production of different prenyl chain length from the intrinsic product, suggesting that this position also plays an important role in product chain-length determination for medium-chain cis-prenyltransferases.

The direct oxygen-sensor protein from Escherichia coli (Ec DOS) is a heme-based signal transducer protein responsible for phosphodiesterase (PDE) activity. We investigated the redox-dependent conformational changes of the sensor domain (Ec DOSH) of Ec DOS by using ultraviolet resonance Raman (UVRR) spectroscopy excited at 229 and 244 nm in combination with site-directed mutagenesis. The UVRR-difference spectra (reduced–oxidized) of the wild-type (WT) revealed several features for the Trp and Tyr bands, indicating that these residues undergo environmental changes. Some of these features were assigned to Trp53, Tyr126, and Tyr80, which are located near the 2-vinyl, 4-vinyl, and propionate side chains of heme, respectively. Other changes of UVRR spectra were assigned to Tyr55, Trp110, and Tyr125, which are located near the surface of Ec DOSH, implying that this protein undergoes global conformational changes upon redox change of heme. Furthermore, the mutation of Tyr55 in the full-length protein (Ec DOS), which is located in heme proximal side near Trp53, perturbed the PDE activity compared with WT, demonstrating the importance of Tyr55 for the catalytic reaction. Finally, visible RR spectra of Ec DOSH showed significant changes in vibrations of heme peripheral groups upon a redox change. Thus, our results strongly suggest that heme redox changes induce structural changes of the heme side chains, and then are communicated to the surface of the sensor domain through Trp53, Tyr126, and Tyr80, resulting in global changes of the protein conformation. Copyright © 2008 John Wiley & Sons, Ltd.

BACKGROUND Hyponatremia has been shown to be a prognostic factor in heart failure (HF) with preserved ejection fraction (HFpEF). Serum sodium (sNa) cut-off, however, is not defined in HFpEF. Therefore, we investigated the relationship between sNa and HF-related events (cardiovascular death and hospitalization for HF decompensation) in HFpEF patients. METHODS AND RESULTS We assessed cardiac function using echocardiography and measured sNa in HFpEF patients with New York Heart Association class II (n=321) or III (n=84) in a compensated condition after implementing medical therapy for HF. During a mean follow-up of 27 months, 73 patients developed HF-related events. On multivariate Cox hazard analysis including established predictors in HF, sNa level as a continuous variable was identified as an independent predictor for HF-related events in HFpEF (per 1.0 mmol/L: HR, 0.93; 95% CI: 0.87-0.98; P

BACKGROUND Endothelial dysfunction plays a crucial role in heart failure (HF), but the association between peripheral microvascular endothelial function assessed by reactive hyperemia peripheral arterial tonometry (RH-PAT) and prognosis remains unknown in HF with reduced left ventricular (LV) ejection fraction (HFREF). We prospectively investigated the association between peripheral microvascular endothelial function and HF-related near-future cardiovascular outcomes in HFREF patients. METHODS AND RESULTS The 362 HFREF patients (LVEF

A boy with Duchenne muscular dystrophy was admitted to our hospital due to a transient loss of consciousness. Transthoracic echocardiography revealed left ventricular (LV) dilatation and diffuse hypokinesis of the LV wall. The LV wall was thin, and both non-compaction of the LV wall and marked thinning of the posterior LV wall resulting from a lesion were observed. The plasma B-type natriuretic peptide (BNP) level ultimately increased to 7,795 pg/mL, and the patient died of cardiac arrest following ventricular tachycardia. Severe heart failure, a critical condition, and thinning of the LV wall may have contributed to the markedly high plasma BNP level in this case.

Risk stratification of chronic kidney disease (CKD) is clinically important because such patients are at high risk of cardiovascular events. Although reactive oxygen species (ROS) are reported to be closely associated with the pathophysiology of CKD, there are few useful ROS biomarkers known for CKD patients. Hence, our objectives in this study were to investigate whether serum derivatives of reactive oxygen metabolites (DROM), a novel biomarker of ROS, is involved in the pathophysiology of CKD (case-control study), and is a significant predictor of future cardiovascular events in CKD patients (follow-up study). Patients with suspected coronary artery disease (CAD) were enrolled and underwent coronary angiography. Patients with CKD (estimated glomerular filtration ratio

Background Reactive oxygen species ( ROS ) are associated with development of coronary artery disease ( CAD ). However, there's no useful biomarker of ROS in CAD . Methods and Results We recruited 395 consecutive CAD patients who were performed coronary angiography (262 male and 133 female, age 70.2±10), and we measured serum derivatives of reactive oxidative metabolites ( DROM ) were measured. Two hundred twenty‐seven non‐ CAD patients were also enrolled. We performed follow‐up study in these 395 CAD patients and case‐control study after risk factor and 1:1 pair matching (both, n=163). As subgroup analysis, DROM were also measured at the aortic root and the coronary sinus in 59 CAD patients. DROM were significantly higher in CAD patients (n=163, median [inter‐quartile range, IQR ]=338 [302 to 386]) than in risk factor‐matched non‐ CAD patients (n=163, 311 [282 to 352.5], effect size=0.33, P CAD patients, 83 cardiovascular events were recorded. Kaplan‐Meier analysis showed a higher probability of cardiovascular events in the high‐ DROM group (>346 U.CARR) than in the low‐ DROM group (≤346 U.CARR) ( P =0.001 [log‐rank test]). Multivariate Cox hazard analysis identified ln‐ DROM as an independent predictor for cardiovascular events (hazard ratio: 10.8, 95% confidence interval: 2.76 to 42.4, P =0.001). The transcardiac gradient of DROM was significantly higher in CAD patients than in non‐ CAD patients (−2.0 [−9.0 to 9.0] versus 8 [−8.0 to 28.3], effect size=0.21, P =0.04), indicating that DROM production in coronary circulation is associated with development of CAD . Conclusion DROM are increased in CAD patients and associated with future cardiovascular events. DROM might provide clinical benefits for risk stratification of CAD . Clinical Trial Registration URL: http://www.umin.ac.jp/ctr/ . Unique identifier: UMIN000012990.

Heme-regulated eIF2alpha kinase [heme-regulated inhibitor (HRI)] plays a critical role in the regulation of protein synthesis by heme iron. The kinase active site is located in the C-terminal domain, whereas the N-terminal domain is suggested to regulate catalysis in response to heme binding. Here, we found that the rate of dissociation for Fe(III)-protoporphyrin IX was much higher for full-length HRI (1.5 x 10(-)(3) s(-)(1)) than for myoglobin (8.4 x 10(-)(7) s(-)(1)) or the alpha-subunit of hemoglobin (7.1 x 10(-)(6) s(-)(1)), demonstrating the heme-sensing character of HRI. Because the role of the N-terminal domain in the structure and catalysis of HRI has not been clear, we generated N-terminal truncated mutants of HRI and examined their oligomeric state, heme binding, axial ligands, substrate interactions, and inhibition by heme derivatives. Multiangle light scattering indicated that the full-length enzyme is a hexamer, whereas truncated mutants (truncations of residues 1-127 and 1-145) are mainly trimers. In addition, we found that one molecule of heme is bound to the full-length and truncated mutant proteins. Optical absorption and electron spin resonance spectra suggested that Cys and water/OH(-) are the heme axial ligands in the N-terminal domain-truncated mutant complex. We also found that HRI has a moderate affinity for heme, allowing it to sense the heme concentration in the cell. Study of the kinetics showed that the HRI kinase reaction follows classical Michaelis-Menten kinetics with respect to ATP but sigmoidal kinetics and positive cooperativity between subunits with respect to the protein substrate (eIF2alpha). Removal of the N-terminal domain decreased this cooperativity between subunits and affected the other kinetic parameters including inhibition by Fe(III)-protoporphyrin IX, Fe(II)-protoporphyrin IX, and protoporphyrin IX. Finally, we found that HRI is inhibited by bilirubin at physiological/pathological levels (IC(50) = 20 microM). The roles of the N-terminal domain and the binding of heme in the structural and functional properties of HRI are discussed.

Neuronal PAS domain protein 2 (NPAS2) is a circadian rhythm-associated transcription factor with two heme-binding sites on two PAS domains. In the present study, we compared the optical absorption spectra, resonance Raman spectra, heme-binding kinetics and DNA-binding characteristics of the isolated fragment containing the N-terminal basic helix–loop–helix (bHLH) of the first PAS (PAS-A) domain of NPAS2 with those of the PAS-A domain alone. We found that the heme-bound bHLH-PAS-A domain mainly exists as a dimer in solution. The Soret absorption peak of the Fe(III) complex for bHLH-PAS-A (421 nm) was located at a wavelength 9 nm higher than for isolated PAS-A (412 nm). The axial ligand trans to CO in bHLH-PAS-A appears to be His, based on the resonance Raman spectra. In addition, the rate constant for heme association with apo-bHLH-PAS (3.3 × 107 mol−1·s−1) was more than two orders of magnitude higher than for association with apo-PAS-A (

The heme-regulated phosphodiesterase from Escherichia coli (Ec DOS), which is a heme redox-dependent enzyme, is active with a ferrous heme but inactive with a ferric heme. Global structural changes including axial ligand switching and a change in the rigidity of the FG loop accompanying the heme redox change may be related to the dependence of Ec DOS activity on the redox state. Axial ligands such as CO, NO, and O2 act as inhibitors of Ec DOS because they interact with the ferrous heme complex. The X-ray crystal structure of the isolated heme-bound domain (Ec DosH) shows that Leu99, Phe113 and Leu115 indirectly and directly form a hydrophobic triad on the heme plane and that they should be located at or near the ligand access channel of the heme iron. We generated L99T, L99F, L115T, and L115F mutants of Ec DosH and examined their physicochemical characteristics, including auto-oxidation rates, O2 and CO binding kinetics, and redox potentials. The Fe(III) complex of the L115F mutant was unstable and had a Soret absorption spectrum located 5 nm lower than those of the wild-type and other mutants. Auto-oxidation rates of the mutants (0.049-0.33 min(-1)) were much higher than that of the wild-type (0.0063 min(-1)). Furthermore, the redox potentials of the former three mutants (23.1-34.6 mV versus SHE) were also significantly lower than that of the wild-type (63.9 mV versus SHE). Interaction between O2 and the L99F mutant was different from that in the wild-type, whereas CO binding rates of the mutants were similar to those of the wild-type. Thus, it appears that Leu99 and Leu115 are critical for determining the characteristics of heme iron. Finally, we discuss the roles of these amino-acid residues in the heme electronic states.

Recent studies have revealed a new class of heme enzymes, the heme-based sensors, which are able to turn on or off cellular signal transduction pathways in response to environmental changes. One of these enzymes is the heme-regulated phosphodiesterase from Escherichia coli (EcDOS). This protein is composed of an N-terminal heme-containing PAS domain and a C-terminal functional domain. PAS is an acronym formed from the names of the Drosophila period clock protein (PER), vertebrate aryl hydrocarbon receptor nuclear translocator (ARNT), and Drosophila single-minded protein (SIM). The heme cofactor in its PAS domain can act as a sensor of the cellular redox state that regulates the adenosine 3',5'-cyclic monophosphate (cAMP) phosphodiesterase activity. The crystal structures of its heme-containing PAS domain have helped clarify how the heme redox-dependent structural changes initiate intramolecular signal transduction. Here, we review recent findings on the structure-function relationships of EcDOS.

Heme-regulated phosphodiesterase from Escherichia coli (DOS Ec ) catalyzes the hydrolysis of cyclic AMP (cAMP) in vitro and is regulated by the redox state of the bound heme. Changes in the redox state result in alterations in the three-dimensional structure of the enzyme, which is then transmitted to the functional domain to switch catalysis on or off. Because DOS Ec was originally cloned from E. coli genomic DNA, it has not been known whether it is actually expressed in wild-type E. coli . In addition, the turnover number of DOS Ec using cAMP as a substrate is only 0.15 min −1 , which is relatively low for a physiologically relevant enzyme. In the present study, we demonstrated for the first time that the DOS Ec gene and protein are expressed in wild-type E. coli , especially under aerobic conditions. We also developed a DOS Ec gene knockout strain (Δ dos ). Interestingly, the knockout of dos caused excess accumulation of intracellular cAMP (26-fold higher than in the wild-type strain) under aerobic conditions, whereas accumulation of cAMP was not observed under anaerobic conditions. We also found differences in cell morphology and growth rate between the mutant cells and the wild-type strain. The changes in the knockout strain were partially complemented by introducing an expression plasmid for dos . Thus, the present study revealed that expression of DOS Ec is regulated according to environmental O 2 availability at the transcriptional level and that the concentration of cAMP in cells is regulated by DOS Ec expression.

Neuronal PAS domain protein 2 (NPAS2) is an important transcription factor associated with circadian rhythms. This protein forms a heterodimer with BMAL1, which binds to the E-box sequence to mediate circadian rhythm-regulated transcription. NPAS2 has two PAS domains with heme-binding sites in the N-terminal portion. In this study, we overexpressed wild-type and His mutants of the PAS-B domain (residues 241-416) of mouse NPAS2 and then purified and characterized the isolated heme-bound proteins. Optical absorption spectra of the wild-type protein showed that the Fe(III), Fe(II) and Fe(II)-CO complexes are 6-co-ordinated low-spin complexes. On the other hand, resonance Raman spectra indicated that both the Fe(III) and Fe(II) complexes contain mixtures of 5-co-ordinated high-spin and 6-co-ordinated low-spin complexes. Based on inverse correlation between nu(Fe-CO) and nu(C-O) of the resonance Raman spectra, it appeared that the axial ligand trans to CO of the heme-bound PAS-B is His. Six His mutants (His266Ala, His289Ala, His300Ala, His302Ala, His329Ala, and His335Ala) were generated, and their optical absorption spectra were compared. The spectrum of the His335Ala mutant indicated that its Fe(III) complex is the 5-co-ordinated high-spin complex, whereas, like the wild-type, the complexes for the five other His mutants were 6-co-ordinated low-spin complexes. Thus, our results suggest that one of the axial ligands of Fe(III) in PAS-B is His335. Also, binding kinetics suggest that heme binding to the PAS-B domain of NPAS2 is relatively weak compared with that of sperm whale myoglobin.

Heme-regulated phosphodiesterase from Escherichia coli (Ec DOS) is a novel PAS heme-sensor enzyme. Ec DOS is active in the Fe2+ heme-bound form, whereas it is inactive in the Fe3+ heme-bound form. To elucidate the mechanism of the redox-dependent heme-regulated catalysis, we examined spectroscopic and functional characters of site-directed and deletion mutant proteins. We also determined crystal structures of the wild type enzyme under various conditions. In this review, we summarized findings about heme-sensor proteins, PAS domain and phosphodiesterase in general and structure and function relationships of Ec DOS specifically.

In haem-regulated phosphodiesterase (PDE) from Escherichia coli (Ec DOS), haem is bound to the PAS domain, and the redox state of the haem iron regulates catalysis by the PDE domain. We generated mutants of Asp40, which forms a hydrogen bond with His77 (a proximal haem axial ligand) via two water molecules, and a salt bridge with Arg85 at the protein surface. The redox potential of haem was markedly increased from 67 mV vs. the standard hydrogen electrode in the wild-type enzyme to 95 mV and 114 mV in the Ala and Asn mutants, respectively. Additionally, the auto-oxidation rate of Ec DOS PAS was significantly increased from 0.0053 to 0.051 and 0.033 min−1, respectively. Interestingly, the catalytic activities of the Asp40 mutants were abolished completely. Thus, Asp40 appears to play a critical role in the electronic structure of the haem iron and redox-dependent catalytic control of the PDE domain. In this report, we discuss the mechanism of catalytic control of Ec DOS, based on the physico-chemical characteristics of the Asp40 mutants.

PAS domains, which have been identified in over 1100 proteins from all three kingdoms of life, convert various input stimuli into signals that propagate to downstream components by modifying protein-protein interactions. One such protein is the Escherichia coli redox sensor, Ec DOS, a phosphodiesterase that degrades cyclic adenosine monophosphate in a redox-dependent manner. Here we report the crystal structures of the heme PAS domain of Ec DOS in both inactive Fe3+ and active Fe2+ forms at 1.32 and 1.9 A resolution, respectively. The protein folds into a characteristic PAS domain structure and forms a homodimer. In the Fe3+ form, the heme iron is ligated to a His-77 side chain and a water molecule. Heme iron reduction is accompanied by heme-ligand switching from the water molecule to a side chain of Met-95 from the FG loop. Concomitantly, the flexible FG loop is significantly rigidified, along with a change in the hydrogen bonding pattern and rotation of subunits relative to each other. The present data led us to propose a novel redox-regulated molecular switch in which local heme-ligand switching may trigger a global “scissor-type” subunit movement that facilitates catalytic control.

Background: Long pentraxin 3 (PTX3) is known to be produced by various types of cell through a lot of inflammatory cytokines such as tumor necrosis factor (TNF)-α. PTX3 is therefore used as an inflammatory marker for acute myocardial infarction and heart failure (HF). However, the precise mechanism of PTX3 in the exacerbation of HF with normal l ejection fraction (HFNEF) remains to be elucidated. Methods: (1) Dahl salt-sensitive hypertensive rats (DS rats), which are the useful model of HFNEF, fed high-salt (HS) DS rats progressively develop hypertension and exhibit LV hypertrophy (LVH) a, followed by HFNEF. In the present study, we used HS loaded DS rats at the age of 15 and 20 weeks to elucidate the mechanism of transition from hypertensive LVH to HFNEF. (2) We examined the levels of PTX3 at the aortic root and the coronary sinus in HFNEF patients who received CAG during the study period. Results: (1) 20-week-old HS loaded DS rats were characterized by prominent LVH and marked LV relaxation abnormality but by the preserved LV systolic function. Compared with control DS rats fed low-salt (LS) diet, LV macrophage infiltration and interstitial fibrosis as shown by histological examination were already significantly increased at the stage of LVH, and remained increased up to the stage of HFNEF (P Conclusion: PTX3 produced from macrophage, rather than from cardiac fibroblasts, might be associated with transition from LVH to HFNEF. Taken together with human data, PTX3 could be a useful marker for discriminating HFNEF from LVH.

Background: Left ventricular (LV) diastolic dysfunction (DD) and vascular functions including peripheral endothelial function play an important role in the pathogenesis of heart failure (HF) with preserved LV ejection fraction (EF) (HFPEF). Hypertension is the most important risk factor in HFPEF and the increased workload caused by hypertension results in LV pathological hypertrophy and LVDD. However, the importance of endothelial function in hypertensive patients with LVDD or HFPEF remains yet undetermined. We investigated the association between peripheral endothelial function, LVDD, and HFPEF in hypertensive patients. Methods and Results: We evaluated cardiac function by echocardiography measuring the ratio of early transmitral flow velocity to tissue Doppler early diastolic mitral annular velocity (E/e’) and LVEF. We also noninvasively assessed peripheral endothelial function by reactive hyperemia-peripheral arterial tonometry (RH-PAT) as the RH-PAT index(RHI) in 405 hypertensive patients with preserved LVEF (LVEF>50%), comprising 180 HFPEF and 225 non-HF patients (LVDD; E/e’>15, non-HF with LVDD; n=98, non-HF without LVDD; n=127). RHI negatively correlated with E/e’ (r=-0.24, P Conclusions: RHI was independently associated with the presence of HFPEF and LVDD in hypertensive patients with preserved LVEF. Endothelial dysfunction in microcirculation could play a crucial role in the pathogenesis of LVDD and HFPEF in hypertensive patients.