bluuebunny/crossref_metadata_2025 huggingface dataset:
PS-P04-10: IMPACT OF BLOOD PRESSURE VARIABILITY ON THE RISK OF STROKE AND PREMATURE DEATH
Authors: Courcoul Leonie , Barbieri Antoine , Woodward Mark , Jacqmin-Gadda Helene , Tzourio Christophe
Published: January 2023

Objective:

Visit-to-visit blood pressure variability (BPV) has been identified in several recent studies as a risk factor for stroke, independently of the level of BP. If true, this finding could help to identify patients at higher risk of stroke and to develop new preventive strategies. However, studies on BPV are exposed to important methodological challenges including conditioning on the future, which violates the assumption of the Cox model, using endogenous variables like the expected BP and its variance, and not handling properly the measurement error. Our objective was to address these issues by designing a joint model with heterogeneous variance for the repeated measures of BP, allowing us to analyze the association between BPV and the risk of stroke and the competing risk of death.

Design and Method:

We used data from PROGRESS (Perindopril Protection against Stroke Study), a placebo-controlled BP-lowering randomized trial for the prevention of stroke recurrence in 6105 patients with a previous history of stroke or TIA. During 5 years of follow-up, BP was measured 12 times, and 727 strokes and 438 non-stroke deaths were observed. We designed an innovative and dedicated joint model combining a linear mixed model, including a subject-specific random effect for the residual variance and two proportional hazard models for the two competing risks of stroke and death. In this model, the risk of each event may depend on the current value and current slope of the individual expected trajectory of BP as well as the intra-subject residual variance.

Results:

In separate analysis of each arm of the trial, we found that the risk of stroke increased with the current level of BP (HR = 1.13 per 5 mmHg, 95%CI = [1.08; 1.17], p-value < 0.001) but was not associated with its intra-individual variability (HR = 0.89, 95%CI = [0.71; 1.12], p-value = 0.317). Conversely, the risk of non-stroke death increased with the intra-individual variability of BP (HR = 1.43, 95%CI = [1.08; 1.88], p-value = 0.011) but not with its current level (HR = 1.03, 95%CI = [0.97; 1.09], p-value = 0.317). Results were similar on the complete sample adjusting for the randomized treatment.

Conclusion:

Using appropriate modeling accounting for measurement error and limiting selection and other potential biases, we found a differential effect of expected BP and BPV on the risk of stroke and of death. These results, if confirmed and enlarged to other populations, would have important consequences to design efficient and individualized preventive strategies.
PS-P05-7: WEIGHT OF EVIDENCE OF LONG-TERM EFFECTS OF HEAVY METALS ON THE CARDIOVASCULAR SYSTEM AT A POPULATION LEVEL
Authors: Saadeh Rami , YoussefAgha Ahmed , Piatt Jennifer , Klaunig James
Published: January 2023

Background/Objectives:

Heavy metals chronic exposure is a major contributor to cardiovascular diseases (CVD) at the population level, and regions with high levels of heavy metals contamination showed increased rate of mortality rate from cardiovascular diseases. Among different heavy metals; Arsenic, Lead, Mercury, and Cadmium are the most toxic to human health.

The purpose of the study was to weigh the amount of evidence presented in the literature for an association between each heavy metal and five cardio-vascular diseases: coronary heart disease, cerebrovascular disease, peripheral arterial disease, hypertension, and atherosclerosis.

Methods:

Literature review was done. PubMed, google scholar, and science direct were searched using keywords cardiovascular diseases and 4 heavy metals, separate and all together. Irrelevant articles were excluded, and only relevant epidemiological articles were included. Bradford Hill criteria of causality was used to assess the association of each of the heavy metals with the five CVD.

Results:

The results of recent studies showed that the correlation between the four heavy metals and CVD are inconsistent with those of earlier studies and the general picture of the correlation between exposure to heavy metals and CVD presented by these studies is either inconsistent or not conclusive.

Conclusion:

Among the nine-criterion mentioned by Sir Bradford Hill, this study was only compatible with two out of nine criterions mentioned. The first criterion is the strength of the association, and the second criterion is the consistency of the association. Other criterions are not compatible with this study.

Although this study tried to examine the effect of 4 heavy metals on CVD, most studies linked only 1 heavy metal to 1 or 2 CVD, due to the interactive effects of heavy metals, and the absence of clear understanding to the mode of action. Nonetheless, there is a great need to do further research and explore more about the effect of these 4 metals on CVD.
PS-P06-6: GENDER DIFFERENCES ON DETERMINANTS OF GOOD KNOWLEDGE ON RECOGNITION OF STROKE SYMPTOMS AND ACTIONS DURING A STROKE AMONG HYPERTENSIVE PATIENT
Authors: Ching Siew Mooi , Chia Yook Chin , Devaraj Navin Kumar , Lim Hooi Min , Beh Hooi Chin
Published: January 2023

Early recognition and action during a stroke are extremely important to improve stroke prognosis. This study aimed to examine the gender difference in the KAP of stroke in hypertensive patients.

This was a cross-sectional study conducted on adults 18 years and older who participated in a community blood pressure (BP) screening program in Malaysia during MMM2017. The questionnaire was derived based on the acronym F.A.S.T recommended by the American Stroke Association and two common symptoms of middle cerebral artery occlusion (paralysis of one side of the body and numbness or tingling sensation of one side of the body). Using multivariate logistic regression, the determinants of recognizing stroke symptoms and knowing what to do during a stroke in female and male hypertensive patients were identified.

Among 1019 hypertensive patients, 85.% of women versus 84% of men were able to recognise symptoms of stroke (p = 0.564) but the prevalence of good knowledge of actions to be taken by being hospitalised within 4.5 hours during a stroke was 74% among women vs. 80% in men (p = 0.05). Using multivariate logistic regression, the determinants for recognising stroke symptoms among male hypertensive patients were age more than 45 year old(OR: 2.670,p = 0.016) compared with those aged below 45, symptom of body paralysis (OR: 9.37,p = 0.001) and symptom of numbness of the body(OR:6.177,p < 0.001);whereas in female hypertensive patient were symptoms of body paralysis (OR:26.582,p < 0.001), being Malay (OR:2.306, p = 0.034) vs. non-Malay and age more than 45 year old(OR:2.706,p = 0.024) compared to those under 45 years old. The determinants of good knowledge on actions to be taken during a stroke among male hypertensive patient were symptoms of body paralysis (OR:8.25,p = 0.031) compared to female respondents were symptom of body paralysis (OR:5.603,p < 0.001),symptom of slurred speech (OR:5.597,p = 0.003) and diastolic BP (OR:0.962, p = 0.005).

Men know better what to do during a stroke. Symptoms of body paralysis was the most common stroke symptoms recognized by all hypertensive patients regardless in order to take further action. Clinicians need to be aware of these findings, and more action needs to be taken to educate female hypertensive patients about other stroke symptoms so that further action can be taken.
PS-R01-4: SAME, SAME BUT DIFFERENT: WHY ABSOLUTE BLOOD PRESSURE VALUES DO NOT ALWAYS PROVIDE AN ABSOLUTELY TRUE PICTURE
Authors: Oh Tania Wei Ling , Mondry Adrian
Published: January 2023

A 45 year old woman was hospitalised for acute, symptomatic Dengue Fever with thrombopenia. Besides body ache and pruritus, she complained of non- postural giddiness. Her blood pressure throughout was in the hypotensive range, hovering between 80- 90 mm Hg systolic and 45–65 mm Hg diastolic. This raised concerns for Dengue- virus mediated endothelial dysfunction and shock. Symptomatic swelling of hands and feet were suspicious for capillary leak syndrome, which would be a relative contraindication for crystalline volume substitution. At the same time, however, the heart rate remained bradycardic, ranging from 52- 74 beats per minute despite febrile temperatures. With incipient thrombocyte recovery, patient requested discharge but agreed to outpatient follow up.

In clinic, she revealed that she had had known low blood pressure and bradycardic heart rate for all her adult life. She had been a competitive swimmer in her youth with up to 10 weekly training sessions until age 17. Given thyroid function and cortisol levels were normal, we carried out two non- invasive cardiac assessments using the NICAs device (NIMedical), which evaluates cardiac function via bioimpedance cardiography. At rest, NICAs showed hypotension (86/47 mm Hg), bradycardia 58 bpm and subnormal cardiac output (3.8 L/min) and cardiac index (2.4 L/min/m2). NICAs was repeated after brief aerobic exercise (50 squats, 20 push-ups, 30 straight leg raises). This brought heart rate into the low normal range, averaging at 61bpm. Systolic blood pressure could be normalised to 92 mm Hg, but diastolic pressure remained low at 51 mm Hg. At the same time, cardiac output and cardiac index moved into the normal range at 4.0 L/min and 2.6 L/min/m2, respectively. While still suffering from mild giddiness, symptoms improved on exercise.

While the measured absolute blood pressure values were in the hypotensive range, the patient was haemodynamically stable. The improved cardiac parameters after exercise and her personal exercise history suggest a permanent set-point to lower sympathetic drive. As such, medical alerts for this patient needed to be individually adjusted, and a personalised approach to treatment is likely to yield better outcomes. The case serves as a reminder that our guidelines are based on studies on large populations, while our patients need to be assessed as individuals if we aim to provide optimal care.
PS-R01-6: SUCCESSFUL ANTIHYPERTENSIVE TREATMENT USING SACUBITRIL/VALSARTAN ALONE IN A PATIENT WITH OBSTRUCTIVE SLEEP APNEA SYNDROME
Authors: Sugihara Ryuta , Sugihara Yuusaku , Sugihara Itsuko
Published: January 2023

Obstructive sleep apnea syndrome (O-SAS) is one of the emerging non-communicable health hazards with high rates of morbidity and mortality. O-SAS is known to induce excessive activity of sympathetic nervous system and result in secondary hypertension. Although continuous positive airway pressure (CPAP) is the first line therapy for O-SAS, its poor adherence induces resistant hypertension. Sacubitril/valsartan has a sympatho-inhibitory effect and may contribute to antihypertensive therapy in patients with O-SAS. We present a successful case of a 39-year-old male with O-SAS using sacubitril/valsartan alone. He visited our clinic with severe hypertension noted during a routine health checkup. He was a heavy drinker, resulted in high body mass index. His family members worried about his loud snoring and witnessed apnea during sleep. The polysomnography showed a markedly increased apnea-hypopnea index of 37, which led to a diagnosis of severe obstructive sleep apnea. At first, we started treatment with CPAP and azilsartan, but he stopped CPAP due to excessive drinking. Then, we switched azilsartan to sacubitril/valsartan, and succeeded in improving his blood pressure.

This case indicates that sacubitril/valsartan may be an effective therapy for uncontrollable hypertenstion due to O-SAS even without CPAP.
PS-R01-8: BLOOD PRESSURE CONTROL WAS IMPROVED BY A CHANGE IN THE TIMING OF ANTIHYPERTENSIVE MEDICATION IN A PATIENT WITH NOCTURNAL HYPERTENSION
Authors: Maruhashi Tatsuya , Higashi Yukihito
Published: January 2023

A 75-year-old-man was referred to our outpatient clinic due to inadequate blood pressure control despite antihypertensive drug treatment. He had dyslipidemia, type 2 diabetes mellitus, and a history of carotid artery stenosis, bilateral central retinal artery occlusion, and lower extremity artery disease. He took losartan potassium 50 mg, hydrochlorothiazide 12.5 mg, cilnidipine 10 mg, and nifedipine CR 40 mg in the morning. Home blood pressure monitoring was performed by using a cuff oscillometric device that could be set to measure blood pressure automatically during sleep. The patient was instructed to measure his morning blood pressure twice after waking, evening blood pressure twice before bedtime, and night-time systolic blood pressure (2:00, 3:00, 4:00, and 5:00 AM) for 7 consecutive days. Mean values were 161 mmHg for morning systolic blood pressure, 134 mmHg for evening systolic blood pressure, and 141 mmHg for night-time systolic blood pressure. After the home blood pressure monitoring, the timing of administration of nifedipine CR 40 mg was shifted from morning dosing to bedtime dosing. One month later, home blood pressure monitoring was performed again using the same schedule as before. Mean values were decreased from 161 mmHg to 135 mmHg for morning systolic blood pressure and from 141 mmHg to 127 mmHg for nigh-time systolic blood pressure. Blood pressure control was improved only by altering the timing of the administration of one antihypertensive drug from morning dosing to bedtime dosing without increasing the dosage of antihypertensive drug or adding a new antihypertensive drug class.
PS-R02-5: A CASE OF CONGESTIVE HEART FAILURE DUE TO HYPERTENSION
Authors: Hashimoto Nonoka , Takase Hiroyuki , Machii Masashi , Hayashi Kazusa , Kin Fumihiko , Nakano Suguru
Published: January 2023

The patient is a 56-year-old man. He was pointed out to have high blood pressure 6 years ago but had not been treated for it. He was hospitalized 4 years ago for heart failure due to atrial fibrillation and underwent catheter ablation therapy. This year, he complained respiratory distress at night and was admitted to the hospital with a diagnosis of congestive heart failure with sinus tachycardia and high blood pressure (height 174 cm, weight 100 kg, blood pressure 178/118 mmHg, pulse 105 bpm). After admission, he was started inhaling oxygen (SpO2 = 97%; nasal oxygen 3L/min) and antihypertensive medications were started with ARNI and calcium channel blocker. His blood pressure decreased, his heart failure improved without the use of diuretics or nitrates, and he no longer required oxygenation. However, because of inadequate pulse control, the calcium channel blocker was changed to beta-blocker, and finally the blood pressure was around 115/80 mmHg and the pulse rate around 85 bpm. At the same time of admission, a close examination of the cause of heart failure was conducted. He smoked 20 cigarettes/day (Ex-smoker from 20 to 35 years old) and still has a drinking habit of 1–2 gou of shochu/day. Blood tests, electrocardiogram, echocardiography, and coronary CT scan were performed to determine that hypertension was the main cause of his heart failure. In order to investigate the involvement of secondary hypertension as a cause of hypertension, examinations including hormone measurements such as catecholamine, aldosterone, renin activity and so on were performed, and a diagnosis of essential hypertension was made. Hypertensive organ damage included hypertensive and arteriosclerotic changes in the fundus and left ventricular hypertrophy. As for cardiovascular disease risk, the patient was male, had a history of non-valvular atrial fibrillation, a smoking history, and moderate Sleep Apnea Syndrome, and belonged to the high-risk group for cerebral cardiovascular disease risk. This patient was able to treat the acute phase of heart failure and maintenance treatment of heart failure only with antihypertensive therapy. The prognosis of hypertensive heart failure is often improved with appropriate antihypertensive treatment.
PS-R02-6: ASSOCIATION BETWEEN DE NOVO TYPICAL CLINICAL-SCENARIO-1 HEART FAILURE AND RENAL ARTERY STENOSIS
Authors: Tasaki Hirofumi , Nagao Shoichi , Yamaguchi Kenji , Furudono Shinnosuke , Ashizawa Naoto , Honda Tomohiro , Akashi Ryohei , Kawano Hiroaki , Maemura Koji
Published: January 2023

A 70-year-old man with sudden onset of dyspnea was referred to our emergency room. Over several years, he had received medical treatments for hypertension, diabetes mellitus, hypercholesterolemia, and obesity, and had a history of heavy smoking until the age of 60. In the past month, systolic blood pressure (BP) increased from around 120/- mmHg to 170/- mmHg or over and exertional dyspnea appeared. On admission, BP was 241/134 mmHg and pulse rate was 140 beats per minute. The chest X-ray showed severe pulmonary edema with butterfly shadow and blood gas analysis revealed the PO2 was 62.2 mmHg, the PCO2 57.7 mmHg, and the pH 7.208 with oxygen inhalation of 12 litters per minute. These were the typical manifestations of clinical scenario (CS) 1 of heart failure (HF). Chemical analysis showed total cholesterol of 240 mg/dl, LDL-cholesterol 159 mg/dl, triglyceride 127 mg/dl, creatinine 1.04 mg/dl, BNP 455.7 pg/ml, and plasma renin activity over 20 ng/ml/hr. Intravenous nitroglycerin, atrial natriuretic peptide and furosemide and noninvasive positive pressure ventilation with oxygen inhalation were consecutively administered. The symptom rapidly resolved thereafter. The left ventricular ejection fraction (EF) of 25–30 % on admission improved to 66 % 5 days later. Abdominal ultrasonography showed an atrophic left kidney (pole-to-pole length 77 mm, resistive index 0.61) compared with the right kidney (pole-to-pole length 96 mm, resistive index 0.71), which suggested the stenosis of the left renal artery and plain computed tomography showed severe calcification of the abdominal aorta and left renal artery. Contrast-enhanced magnetic resonance imaging and diagnostic angiography revealed severe left renal stenosis. There was no significant stenosis in any coronary arteries. After renal revascularization, BP decreased and antihypertensive agents were reduced, and there was no exacerbation of HF afterward. Renovascular hypertension (RH) is a rare disease in daily practice and is recognized as one of the causes of HF with unknown etiology. RH with recurrent exacerbation of HF is the indication for renal revascularization. However, renal artery stenosis is rarely identified even in HFpEF (HF with preserved EF) patients. We experienced a de novo HFpEF case with the typical features of CS 1 of hypertensive emergency complicated by RH with underlying multiple arteriosclerotic diseases. The association between de novo typical CS-1 heart failure of hypertensive emergency and renal artery stenosis might be closer and thus both conditions might more often coexist than expected, and once RH would be identified, the clinical course could be carefully observed thereafter.
PS-R02-9: A PATIENT HAS THE ONLY CONNECTION BETWEEN THE ATRIUM AND VENTRICLE IS AN ACCESSORY PATHWAY
Authors: Ulusan Sebahat , Kuyumcu Mevlut Serdar
Published: January 2023

Case report:

A 62-year-old woman without cardiovascular risk factors such as hypertension, diabetes, smoking and hyperlipidemia was admitted to our cardiology outpatient clinic with palpitations and syncope history. Her electrocardiography showed preexcitation possible posteroseptal accessory pathway. We performed electrophysiological study and found posteroseptal accessory pathway. Tachycardia was not induced in inductions. ERP detected below 240 msn. The area where the shortest V-A interval was detected. His signal was not detected in this region. We determined that it was anatomically far from the AV node. We started radiofrequency ablation and in the 5. second of ablation, we saw that the patient entered 3. degree AV block. Ablation was immediately stopped then sinus rhythm restored in patients qucikly. We search his signal with ablation catheter in right atrium but could not find the signal. We detected that the AV block and accessory pathway were found together in the patient. Since the patient had a history of syncope and ERP < 250 ms was detected, we planned ablation after pacemaker implantation in the patient. We implant DDDR pacemaker and 2 weeks later we make successfully accessory pathway ablation.

Discussion:

In these patients, atrioventricular conduction is done only, and entirely, via the accessory route. A complete block, nodal or sub-Hissian, underlying, will only be visible on the ECG if there is concomitant blockage of the atrioventricular conduction through the accessory pathway. The appearance of atrioventricular conductive disorders after radiofrequency ablation of an accessory pathway, located at a distance from the atrioventricular junction, is not considered by some authors to be a real complication of this procedure. It would rather be a prior conductive disorder having been unmasked following the blockage of the accessory pathway thanks to ablation. It is therefore logical to review the indication for the removal of the accessory pathway in these cases, especially in asymptomatic patients who have never reported a history of palpitation attacks. The only possible tachycardia remains atrial fibrillation on WPW. Only the study of anterograde refractory periods makes it possible to specify the correct indication for ablative treatment. The indication for the implantation of a pacemaker is still appropriate given the risk of progressive and subsequent degeneration of the accessory pathway. In addition, conduction via the accessory pathway, whether continuous or intermittent, would help prolong the longevity of the pacemaker by inhibiting ventricular pacing in the event of atrioventricular conduction through the accessory pathway.
PS-BPB01-5: LOX-1 DEFICIENCY INCREASES RUPTURED ABDOMINAL AORTIC ANEURYSM VIA THINNING OF ADVENTITIAL COLLAGEN
Authors: Takahashi Kayo , Aono Jun , Nakao Yasuhisa , Hamaguchi Mika , Suehiro Chika , Kurata Mie , Kakino Akemi , Sawamura Tatsuya , Inoue Katsuji , Ikeda Shuntaro , Suzuki Jun , Yamaguchi Osamu
Published: January 2023

Objective:

Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a key mediator of inflammation and plays an important role in the pathogenesis of atherosclerosis. Both inflammation and atherosclerosis have been proposed as contributors to abdominal aortic aneurysm (AAA) pathogenesis. However, the role of LOX-1 in AAA pathogenesis remains unknown. We aimed to investigate effects of Olr1, which encodes LOX-1, deletion on angiotensin II (Ang II)-induced AAA in apolipoprotein E knockout (ApoE KO) mice to determine whether LOX-1 deficiency mitigates AAA development.

Design and method:

Either Ang II (1.44 mg/kg/day) or saline (control) was administered continuously through a subcutaneously implanted osmotic pump for 4 weeks in the following 8-week-old male mice: Apoe-/-Olr1+/+
mice (Ang II, n = 40; saline, n = 10), Apoe-/-Olr1+/-
mice (Ang II, n = 30; saline, n = 9), and Apoe-/-Olr1-/-
mice (Ang II, n = 39; saline, n = 9).

Results:

Ang II infusion raised systolic blood pressure by tail-cuff methods in all three groups. Serial non-invasive assessment with ultrasound demonstrated that Ang II gradually increased the maximum area of the suprarenal abdominal aortas leading to AAAs. Although the incidence and expansion rate of AAA were comparable among genotypes, Olr1 deletion significantly increased the severity and rupture rate of AAAs (p = 0.01; incidence of type IV and ruptured AAA was 25% in Apoe-/-Olr1-/-
mice vs 5% in Apoe-/-Olr1+/+
mice). Since atherosclerosis is considered a major contributor to the development of AAAs, the extent of the atheroma burden localized in aneurysmal lesions was evaluated by Oil Red O staining. Olr1 deletion did not decrease atherosclerosis formation in the aneurysmal wall. Interestingly, further histopathological analysis revealed that aneurysmal lesions in LOX-1-deficient mice had fewer fibroblasts and myofibroblasts, as well as thinner adventitial collagen, although the degree of elastin fragmentation or disruption was similar between LOX-1-deficient and control mice. These in vivo data suggested that adventitial fibroblasts play a pivotal role in the increased severity of AAA in LOX-1-deficient mice. To further determine the effects of Olr1 deletion on fibroblast differentiation, fibroblasts were isolated from the adventitia of abdominal aortas harvested from 8-week-old Apoe-/-Olr1+/+
and Apoe-/-Olr1-/-
mice, and then cultured with Ang II stimulation. An in vitro study confirmed that Olr1 deletion completely abolished the Ang II-induced proliferative activity in the aortic adventitial fibroblasts (p = 0.02).

Conclusions:

Olr1 deletion may not mitigate aneurysm development but rather increase the vulnerability to rupture by suppressing adventitial fibroblast proliferation and collagen.
PS-BPB01-9: OMEGA-3 FATTY ACIDS ATTENUATE OXIDATIVE STRESS-INDUCED DNA DAMAGE THROUGH NRF-2 IN VASCULAR ENDOTHELIAL CELLS
Authors: Sakai Chiemi , Ishida Mari , Ishida Takafumi
Published: January 2023

Objective:

Impaired vasodilation is one of the characterizations of hypertension. The effects of the omega-3 fatty acids on blood pressure and vascular function have been examined in many randomized controlled trials. In this study, we investigated the effects of omega-3 fatty acids, particularly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), on DNA damage, which has crucial effects on vascular endothelial cell function.

Design and method:

We examined the effects of EPA and DHA on H2O2-induced DNA damage response in human aortic endothelial cells (HAECs). DNA double-strand breaks (DSBs) were detected by immunofluorescence staining of the phosphorylated form of the histone H2AX (gamma-H2AX).

Results:

EPA and DHA significantly reduced accumulation of DSBs and phosphorylation of ATM, a serine/threonine kinase recruited and activated upon DSBs, in HAECs. Since EPA and DHA attenuated DNA damage without enhancement of DNA repair response, the effects of EPA and DHA on oxidative stress, a DNA damage inducer, were examined. CM-H2DCFDA measurement showed that EPA and DHA significantly reduced intracellular reactive oxygen species. In addition, mRNA levels of antioxidant molecules, such as heme oxygenase-1 and thioredoxin reductase 1, were significantly elevated by EPA and DHA. Silencing the gene encoding nuclear factor erythroid 2-related factor 2 (Nrf2) abrogated these increases. Furthermore, EPA and DHA reduced H2O2-induced senescence-associated beta-galactosidase activity, which was revoked by Nrf2 silencing.

Conclusions:

Our results suggest that EPA and DHA attenuate oxidative stress-induced DNA damage and cell senescence in vascular endothelial cells partly via antioxidant response mediated by Nrf-2. Therefore EPA and DHA may play a role in the prevention of impairment in vasodilation by their genome-protective properties.
PS-BPB03-2: VASORELAXANT-MEDIATED ANTIHYPERTENSIVE EFFECT OF THE SEED AQUEOUS EXTRACT OF AFRAMOMUM PRUINOSUM GAGNEP. (ZINGIBERACEAE)
Authors: Mbuyo Elvine Pami Nguelefack , Mekontchou Larissa Orelle , Tchinda Nidele Lonla , Goumtsa Ariane Falone , Atsamo Albert Donatien , Nguelefack Telesphore Benoit
Published: January 2023

Objective:

Despite the existing therapies, arterial hypertension is still a prevalent and a worldwide major public health concern, prompting the search for new therapeutic alternatives. The present study investigated the antihypertensive and the related mechanisms of action of the seed aqueous extract of Aframomum pruinosum (AEAP), a plant used in Cameroon traditional medicine against cardiovascular ailments.

Design and method:

Male Wistar rats were rendered hypertensive by chronic oral administration of L-NAME (30 mg/kg/day) for three consecutive weeks, and hypertensive rats were selected and treated with L-NAME alone or in combination with AEAP at doses of 75 or 150 mg/kg/day for an additional 3 consecutive weeks. Captopril (25 mg/kg/day, p.o) was used as reference drug. Blood pressure and heart rate were monitored weekly. At the end of the experiment, the heart was harvested, weighed, and sections of cardiac tissue were stained with hematoxylin and eosin for histological analysis. Cumulative concentrations of the aqueous extract (3, 10, 30, 100, and 300 μg/mL) were tested on intact or endothelium-denuded aorta rings precontracted with norepinephrine (NE; 1 μM) or KCl (90 mM) in absence or presence of L-NAME (a nitric oxide synthase inhibitor, 100 μM), indomethacin (a prostaglandin synthesis inhibitor; 10 μM), methylene blue (a guanylate cyclase inhibitor, 1 μM), tetraethylammonium (TEA; a non-specific potassium channel blocker; 5 μM) or glibenclamide (an ATP-sensitive potassium channel blocker; 10 μM). The contractile response to NE, or CaCl2 was also assessed in the presence or absence of the plant extract.

Results:

AEAP completely reversed (p < 0.001) L-NAME-induced elevated systolic and diastolic blood pressure, and increased heart rate. AEAP partially reversed cardiac hypertrophy, leucocyte infiltration, and prevent cardiac necrosis. AEAP induced a concentration-dependent vasorelaxant effect on NE- and KCl-evoked contraction with EC50 values of 13.46 and 49.37 μg/mL, respectively. Removal of the endothelium and the preincubation of aorta rings with L-NAME significantly (p < 0.05) reduced the vasorelaxant effect of the plant extract, while indomethacin and methylene blue had no effect (p > 0.05). Glibenclamide, similar to TEA, significantly (p < 0.001) inhibited the vasorelaxant response to AEAP. Also, the extract significantly (p < 0.01 and p < 0.001) inhibited NE- and CaCl2-induced contraction.

Conclusion:

AEAP possesses antihypertensive effects that are at least partially mediated by its vasorelaxant activity. Nitric oxide release, opening of KATP channels, blockade of alpha-1 receptors, and inhibition of Ca2+ influx account for this vasorelaxant activity.
PS-BPB04-7: PRO-ANGIOGENIC EFFECTS OF LSKL, A THROMBOSPONDIN-1 INHIBITOR, IN HUMAN AND EXPERIMENTAL MODELS OF PREECLAMPSIA
Authors: Marc Casandra , Achille Allahnah , Fregeau Genevieve , Hannou Lydia , Vachon Isabelle , Vaillancourt Cathy , Lavoie Julie , Bertagnolli Mariane
Published: January 2023

Introduction:

Preeclampsia (PE) is a hypertensive disorder of pregnancy characterized by blood pressure above 140/90 mmHg. It accounts for 10% of all pregnancies and is responsible for 25% of preterm births and low birth weight. To date, no medications can treat or prevent PE. We identified an activation of the anti-angiogenic protein thrombospondin-1 (THBS1) in human and experimental preeclampsia. THBS1 is also a main activator of TGF-β. We postulate that THBS1-mediated TGF-β activation may contribute to impair placental angiogenesis during PE.

Objective:

We aimed to test the effects of LSKL, an inhibitor of THBS1-mediated TGF-β activation, on angiogenesis in placentas of a mouse model of PE and in human placental endothelial cells.

Methods:

Peptide LSKL or its control peptide SLLK (280 μM in 0.5 ml saline, s.c.) was administered at gestational day (GD) 14.5 in heterozygous transgenic mice overexpressing human renin and human angiotensinogen and controls (C57BL/6). Placentas (n = 12/group) were removed at GD 18.5. Protein expression of TGF-β1, TGF-β2, total and phosphorylated SMAD2 were assessed by western blot of placental lysates. Placental angiogenesis was assessed by CD31+ immunohistochemistry in placental sections. CD31+ endothelial cells were extracted by magnetic sorting from human placentas (n = 4) of healthy pregnancies. Cells (2–5 passages) were treated with LSKL or SLLK (30 μM) under hypoxia (0.5% vs. 8% O2) or thrombin (10 u/mL) exposure (4 hours). THBS1 expression in placental endothelial cells was assessed by immunofluorescence. Cell angiogenesis was quantified by the number of closed tubes formed on matrigel (per 15,000 cells, 4 hours).

Results:

Maternal LSKL treatment significantly reduced the expression of TGF-β2 (P < 0.01) and SMAD2 phosphorylation (P < 0.05) while improving angiogenesis (P < 0.05) in placentas of transgenic preeclamptic mouse compared to controls. In human placental endothelial cells, both thrombin (P < 0.05) and hypoxia (P < 0.05) significantly increased THBS1 expression. Thrombin (P < 0.01) and hypoxia (P < 0.05) significantly impaired cell angiogenesis on matrigel (P < 0.01), which was prevented by LSKL treatment (P < 0.01). In human cells, LSKL similarly reduced TGB-β2 (p < 0.05) and phospho-SMAD2 (P < 0.05) expressions under thrombin and hypoxia cell exposure.

Conclusion:

Our results describe a significant pro-angiogenic effect of LSKL treatment by regulating THBS1 and TGF-β2 mechanisms in human and experimental models of PE.
PS-BPB04-8: ANGIOTENSIN II FORMULATIONS AND THEIR IMPACT ON CARDAIC, VASCULAR AND INFLAMMATORY MECHANISMS OF HYPERTENSION
Authors: Nosalski Ryszard , Booth Emma , Saju Blessy , Alsheikh Eman , Crespo Eva , Maffia Pasquale , Graham Delyth , Guzik Tomasz
Published: January 2023

Angiotensin II (AngII) is a major effector of the renin-angiotensin system, with a critical role in BP regulation of vascular function, and end-organ damage. AngII infusion, using osmotic minipumps, is the most common animal model of hypertension, recapitulating key vascular and cardiac features of human hypertension. While different formulations of AngII are available, their impact on the key outcomes of the model remains unknown. Accordingly, we compared two most commonly used AngII formulations, and their impact on the key cardiovascular endpoints in a mouse model of hypertension.

Male 12-week old C57BL/6J mice (n = 11–14) underwent Sham, AngII[Val5] or AngII[Ile5] (490 ng/min/kg) treatment for two weeks, using osmotic minipumps. Blood pressure was measured by tail-cuff, vascular function by wire myography, remodelling by picrosirius red staining, mRNA expression using real-time PCR and perivascular (PVAT) inflammation by flow cytometry. Data were analyzed using one-way or RM-two-way ANOVA.

Both AngII formulations caused a significant BP elevation (p < 0.001) in comparison to sham animals, although AngII[Val5] had a more pronounced pressure effect than AngII[Ile5] (171 ± 4 vs 156 ± 6 mmHg, p = 0.03). Similarly, endothelium-dependent relaxation of resistance arteries and aorta was significantly impaired by AngII infusion (p < 0.001), however, a maximum Ach-induced relaxation was significantly lower in AngII[Val5] than in AngII[Ile5]. Both, AngII formulations increased intima-media thickness of the aorta (p < 0.0001), although the effect of AngII[Val5] was ∼2-fold higher than AngII[Ile5] (p = 0.001). While animals infused with AngII[Val5] had enhanced vascular collagen deposition (p < 0.001), this effect was lesser in the AngII[Ile5] group (p = 0.02). The total number of leukocytes infiltrating PVAT was induced by both AngII formulations, however, this effect was less pronounced in AngII[Ile5] (1002 ± 376 vs. 753 ± 242 cell/mg, p = 0.03). The same effect was found among CD11b+ and CD11c+ cells. While a significant increase of CD3+ cells was observed in both AngII formulations. While cardiac hypertrophy was evident in both AngII formulations this effect was significantly lower in AngII[Ile5] (p = 0.03). This was confirmed by cross-sectional area of cardiomyocytes. Histological analysis of heart sections showed a significant effect of both AngII formulation on cardiac fibrosis (p < 0.05). This effect was more pronounced upon AngII[Val5] infusion (p < 0.001) and was associated with higher expression of pro-fibrotic markers.

AngII[Val5] formulation is a more efficient and consistent way to study key cardiovascular endpoints such as BP, inflammation, vascular and cardiac damage compared with AngII[Ile5]. Furthermore, the use of AngII[Val5] leads to a more pronounced phenotype and may be associated with reducing the number of animals needed in experiments.
PS-BPB05-6: TOLVAPTAN INDUCES BODY FLUID LOSS AND ACTIVATION OF RENAL UREA-DRIVEN WATER CONSERVATION IN NORMAL RATS
Authors: Kidoguchi Satoshi , Kitada Kento , Fujisawa Yoshihide , Nakano Daisuke , Yokoo Takashi , Titze Jens , Nishiyama Akira
Published: January 2023

Objective:

We recently reported that body fluid loss models such as high salt-induced natriuresis, impaired urine concentration ability via kidney injury, and enhanced transepidermal water loss due to skin barrier dysfunction exhibit initial water loss and subsequent activation of the water conservation system in multiple organs. This activation of the water conservation system restores or even increases water content at the tissue level, which may be involved in abnormal body fluid distribution in various disease models. The water conservation system consists of renal urea recycling, the energy-intensive nature of hepato-muscular ureagenesis, and suppression of cardiovascular energy expenditure to save energy for ureagenesis. However, it is not clarified whether pharmacological fluid loss by diuretics activates the water conservation system. In this study, we examined the effects of water diuresis by tolvaptan, a selective vasopressin V2 receptor antagonist, on the water conservation system in normal rats.

Design and method:

10-week-old male Sprague Dawley rats were fed with a control diet or a diet containing 0.1% tolvaptan. We measured water intake and urine volume for two weeks and also evaluated time-dependent changes in renal urea recycling and hepato-muscular ureagenesis in these rats. In addition, we also evaluated the effects of tolvaptan on blood pressure, heart rate, and renal sympathetic nerve activity by a radiotelemetry system in separate animals.

Results:

Tolvaptan markedly increased urine volume up to 126.7 ± 25.9 ml on day-1, which gradually decreased as much as one-third (48.1 ± 18.0 ml) on day-7. Tolvaptan significantly decreased body water content and increased plasma albumin concentration, an indicator of oncotic pressure, on day-1, suggesting that tolvaptan decreased body fluid and plasma volume on day-1. These changes in total body water content and plasma albumin concentration were restored on day-7. The restoration of body fluid on day-7 was associated with an increase in urea transporter A1-associated renal urea recycling. On the other hand, tolvaptan did not alter hepato-muscular ureagenesis or cardiovascular energy expenditure throughout the treatment.

Conclusions:

Tolvaptan induces initial body fluid loss by water diuresis and subsequent activation of renal urea recycling to restore the water loss. Activation of the water conservation system in addition to a rise in urine volume may be involved in the unknown distinct effect of the diuretics.
PS-BPB07-1: DEVELOPMENT OF ANTI-THROMBOTIC VACCINE AGAINST HUMAN S100A9
Authors: Shimamura Munehisa , Kaikita Koichi , Hayashi Hiroki , Tsujita Kenichi , Nakagami Hironori , Morishita Ryuichi , Rakugi Hiromi
Published: January 2023

Background & Purposes:

In post-stroke patients, a decreased adherence to antiplatelet drugs is a major challenge in the prevention of recurrent stroke. Previously, we have reported that a newly designed antiplatelet vaccine against S100A9 could prevent thrombosis in the middle cerebral artery in mice by inhibiting S100A9/CD36 signaling in platelets. However, the difference in amino acid sequences in epitopes between mice and humans might be problematic for clinical use. In the present study, we redesigned the S100A9 vaccine for the common sequence in both humans and monkeys and examined its effects in cynomolgus monkeys.

Methods:

First, we screened several candidate epitopes in mice and the selected epitope in human S100A9, which is common in monkey S100A9, was conjugated to Keyhole limpet hemocyanin. A mixture of the vaccine and Alum adjuvant was subcutaneously injected into 4 cynomolgus monkeys, three times every two weeks. Antibody production against human S100A9 was examined by ELISA or western blotting. Anti-thrombotic effects in collagen-induced thrombosis were examined in a mixture of vaccinated serum and fresh whole blood from another cynomolgus monkey using an automated microchip flow-chamber system. Bleeding time was evaluated in two monkeys.

Results:

Based on results of computed epitope prediction, we first assessed whether 2 to 11, 97–106, or 104–113 amino acids, where the amino acid sequence is approximately similar between humans and monkeys, could raise antibodies against S100A9 in mice. Compared with the vaccine for the 2–11 amino acids epitopes, 97–106 amino acids and 104–113 amino acids demonstrated higher antigenicity, indicating that epitopes in the C-terminus are stronger than those in the N-terminus. Based on this result, we selected 102 to 112 amino acids as the epitope for the human S100A9 vaccine. When the monkeys were vaccinated, antibody titers against human S100A9 were significantly increased. Western blotting analysis showed that the produced antibody recognized human S100A9. Although the addition of pre-vaccinated serum into fresh whole blood did not affect thrombus formation, post-vaccination serum inhibited collagen-induced thrombosis formation in two monkeys. Anti-thrombotic effects were partially inhibited by the epitope peptide, indicating the feasibility of neutralizing anti-thrombotic effects by vaccination. Notably, prolongation of bleeding time was not observed in vaccinated monkeys.

Conclusions:

Anti-human S100A9 vaccine revealed anti-thrombotic effects in some monkeys without affecting bleeding time. Although further studies are necessary to increase the effects and warrant the safety aspects, this vaccine could clarify the use of an anti-thrombotic vaccine in clinical settings.
PS-BPB08-2: DEVELOPMENT OF NON-PEPTIDE TYPE OF ANGIOTENSIN II TYPE 1 RECEPTOR LIGAND AS A BIASED AGONIST
Authors: Matsuo Yoshino , Suematsu Yasunori , Morita Hidetaka , Miura Shinichiro
Published: January 2023

Objective:

Peptide type of angiotensin II (Ang II) type 1 (AT1) receptor ligand as a biased agonist antagonized Ang II-stimulated G protein signaling, but stimulated several kinase pathways. Here, we developed non-peptide type of AT1 receptor compound as a biased ligand.

Design and method:

We synthesized three non-peptide type of AT1 receptor ligands (Compound A, B and C) as candidates of biased ligands. These compounds were provided by Daiichi Sankyo Co. LTD. In vitro studies, inositol phosphate (IP) production and extracellular signal-regulated kinase (ERK) 1/2 activation were measured using AT1 receptor overexpressing cell system. We mutated the AT1 receptor (L112A, Q257A, Y292A and N295A receptors) and examined binding modes of receptor-ligand by competition binding study, and subsequently analyzed whether these ligands would induce IP production and ERK1/2 activation.

Results:

Kd of Compound A, B and C to AT1 wild-type receptor by competition binding study were 0.8, 21 and 48 nM, respectively. Kd of Compound A, B and C to L112A receptor were 37, 23 and 31 nM, respectively. Compound B and C decreased and increased IP production, respectively, whereas Compound A did not change IP production. Compound B and C, but not Compound A, activated ERK1/2. L112A had a key role of IP production.

Conclusions:

Compound A, B and C were a neutral antagonist, an inverse agonist, and an agonist with regard to IP production, respectively. On the other hand, Compound B and C, but not Compound A, were agonists to ERK1/2 activation. Thus, we developed non-peptide type of AT1 receptor compound as a biased ligand.
PS-BPB09-2: DEVELOPMENT OF LONG-ACTING HUMAN ADRENOMEDULLIN FC FUSION PROTEINS
Authors: Nagata Sayaka , Yamasaki Motoo , Kuroishi Nobuko , Kitamura Kazuo
Published: January 2023

Background:

Human adrenomedullin (hAM) is a hypotensive peptide hormone with strong anti-inflammatory effects. AM consists of 52 amino acid residues with an amidated C-terminus and a ring structure formed by intramolecular disulfide bonds. Both of these structures are necessary for binding to receptors and to exert their biological functions. AM has also shown therapeutic effects in various animal disease models, including inflammatory bowel disease, ischemic heart disease, sepsis and stroke. However, the short half-life of native AM in the blood required continuous administration for treatment. To solve this, we have developed four human IgG1 and IgG4 Fc fusion proteins containing full-length human AM or AM(6–52).

Methods:

Recombinant Fc-AM derivatives were generated using mammalian cells. After that, we tested the biological activities of Fc-fusion AMs (Fc-AMs) to stimulate the intracellular accumulation of cAMP in HEK-293 cells stably expressing the AM1 receptor. Next, we compared IgG1-AM (6–52) and IgG4-AM (6–52) concentrations in the peripheral blood and tissues of rats after subcutaneous injection. In addition, the antihypertensive effect of Fc-AM was examined on spontaneously hypertensive rats (SHRs) which were given high salt diets.

Results:

We have developed four Fc-AMs, which are long-acting AM derivatives in mammalian cells. Fc-AMs produced in mammalian cells do not require refolding or amidation. Indeed, Fc-AMs could be measured by mAM assay, which recognizes the amidated form, indicating that Fc-AMs produced in mammalian cells were amidated. Fc-AMs, generated in a mammalian cell production system can therefore easily be produced in large amounts with few purification steps. The Fc-AMs stimulated intracellular cAMP production in cultured cells stably expressing the AM type I receptor in vitro. Fc-AM (6–52) induced higher cAMP levels for the receptor than Fc-AM. The pharmacokinetic study was performed using N-terminal deficient AM derivatives, which have strong receptor binding ability. Sufficient concentrations of IgG1-AM (6–52) and IgG4-AM (6–52) were observed in blood 14 days after a single subcutaneous administration. Tissue transfer to the kidney and small intestine was observed after administration of IgG1-AM (6–52) or IgG4-AM (6–52) to rats. In addition, the single subcutaneous administration of IgG4-AM (6–52) suppressed the increase in blood pressure in SHRs which were loaded with high salt diets.

Conclusion:

Fc-AM produced from mammalian cells is easily prepared and seems to be an effective new therapeutic agent for hypertension.
PS-BPB09-8: ANGIOTENSIN RECEPTOR BLOCKER AMELIORATES ELEVATED ARTERIAL BLOOD PRESSURE AND REDUCES BODY MASS IN OLETF RATS FED A HIGH CHOLESTEROL DIET
Authors: Jardines Eira Edith , Rodriguez Ruben , Minas Jacqueline , Nakano Daisuke , Nishiyama Akira , Ortiz Rudy M
Published: January 2023

Objective:

Cardiovascular disease (CVD) is the leading cause of death in the US (1 in 4) and is a primary outcome of Metabolic Syndrome (MetS). MetS affects nearly 40% of adults over 60 and is defined by a cluster of conditions including elevated arterial pressure and excess abdominal fat. A high cholesterol diet (HCD) is known to promote cluster-factor conditions of MetS. Unfortunately, body mass (BM) reduction is often insufficient to reduce hypertension. The first line of pharmaceutical defense is disruption of the renin-angiotensin aldosterone system (RAS). Inappropriate elevation of RAS can be therapeutically attenuated with angiotensin receptor blocker (ARBs), which has been shown to ameliorate other MetS cluster factor conditions. However, the efficacy of ARB on elevated arterial pressure during a HCD remains unclear.

Design:

A cohort of 5-week-old Otsuka Long Evan Tokushima Fatty (OLETF) rats were fed a HCD (2% + 0.5% cholic acid) or chow ad libitium. After 4 weeks, animals were treated with ARB (10 mg olmesartan /kg/d x 3 wks by oral gavage) and assigned to five groups: 1) lean-strain control Long Tokushima Evans Otsuka (LETO; n = 10), 2) OLETF (OLETF; n = 12), 3) OLETF+HCD (n = 12), 4) OLETF+ARB (n = 12) and 5) OLETF+HCD+ARB (n = 12). Systolic blood pressure was monitored chronically by radio telemetery.

Results:

OLETF SBP was 31% (p < 0.05) greater than LETO. HCD did not exacerbate elevated SBP in OLETF. ARB reduced SBP in OLETF + HCD by 11% (p < 0.05) over 30 days and 13% (p < 0.05) overall compared to OLETF. ARB reduced BM by 3% (p < 0.05) in chow-fed and 4% (p < 0.05) in HCD-fed OLETF compared to OLETF. However, there was no difference in BM between OLETF+ARB and OLETF+HCD+ARB. Neither, HCD nor ARB alone reduced abdominal fat in OLETF rats. ARB in OLETF+HCD induced visceral fat loss (37%; p < 0.05) compared to OLETF. ARB administration did not attenuate plasma angiotensin II levels.

Conclusion:

These findings demonstrate that chronic treatment with ARB can effectively ameliorate the MetS-associated hypertension despite the additional insult of a HCD.