Because of the high mortality from myocardial infarction and stroke, there is a great demand for finding novel methods of diagnosis, prevention and treatment of these diseases. Most of the current tests measure important determinants of thrombosis such as platelet function, coagulation and fibrinolysis in isolation; therefore, a global test measuring the actual thrombotic status would be more useful in clinical conditions. We obtained considerable experience by using the global thrombosis test, which determines the actual thrombotic status by taking into account the measured platelet reactivity, coagulation and fibrinolytic activities. In animal experiments, we found significant correlation between the ex vivo global thrombosis test measurements and the in vivo thrombotic status. The published evidence for the benefit of an antithrombotic diet with regular physical exercise is also described.

Junichiro Yamamoto*,1, Nobutaka Inoue2, Kazunori Otsui3, Hideo Ikarugi4, Muneshige Shimizu5, Shinji Yamamoto6, Masahiro Murakami7, Yoshinobu Ijiri8 & Kjell S Sakariassen9 1Kobe Gakuin University, Kobe 651–2180, Japan 2Department of Cardiovascular Medicine, Kobe Rosai Hospital, Kobe 651–0053, Japan 3Department of General Internal Medicine, Kobe University Hospital, Kobe 650–0017, Japan 4School of Economics & Management, University of Hyogo, Kobe 651–2197, Japan 5Department of Fisheries, Faculty of Marine Science & Technology, Tokai University, Shizuoka 424–8610, Japan 6Faculty of Sport Sciences, Nihon Fukushi University, Aichi 470–3295, Japan 7Laboratory of Pharmaceutics, Faculty of Pharmacy, Osaka Ohtani University, Osaka 584–8540, Japan 8Department of Health & Nutrition, Faculty of Health & Nutrition, Osaka Shoin Women’s University, Osaka 577–8550, Japan 9Str. Campo e Zampe 12, I–13900, Biella, BI, Italy *Author for correspondence: yamamoto@nutr.kobegakuin.ac.jp

Future Science OAVol. 5, No. 2 EditorialOpen AccessShear rate-dependent thrombosis/fibrinolysis tests using non-anticoagulated blood could be useful in the prevention of thrombotic disordersJunichiro Yamamoto, Yoshinobu Ijiri, Muneshige Shimizu, Shinji Yamamoto, Hideo Ikarugi & Kjell S SakariassenJunichiro Yamamoto*Author for correspondence: E-mail Address: yamamoto@nutr.kobegakuin.ac.jp Kobe Gakuin University (Professor Emeritus), Kobe, JapanSearch for more papers by this author, Yoshinobu Ijiri Faculty of Health & Nutrition, Osaka Shoin Women's University, Osaka, JapanSearch for more papers by this author, Muneshige Shimizu Otsuka Pharmaceutical Company, Saga, JapanSearch for more papers by this author, Shinji Yamamoto Faculty of Sport Sciences, Nihon Fukushi University, Aichi, JapanSearch for more papers by this author, Hideo Ikarugi School of Economics, University of Hyogo, Kobe, JapanSearch for more papers by this author & Kjell S Sakariassen Biella, BI, ItalySearch for more papers by this authorPublished Online:19 Nov 2018https://doi.org/10.4155/fsoa-2018-0092AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinkedInReddit Keywords: dietexercisefibrinolysisnative bloodplatelet functionshear rate-dependent thrombosisthrombolysisthrombusPrevention of thrombotic disorders such as cardiovascular disease and stroke is an urgent and important task for society. Prevention by suitable diet and exercise is recommended by government guidelines in many countries. In order to get useful and practical results by the recommendations, tests employed to assess the thrombotic status of patients, their quality of diet and their exercise levels have conclusive importance.Assessment of thrombotic status has been performed over the years by quantifying thrombotic factors and by measuring function using anticoagulated blood samples. However, these approaches do not seem to be successful in assessing thrombotic status. This may be due in part to the belief that adding calcium to anticoagulated blood can restore properties of the original native blood and that individual quantification of thrombotic factors can reflect properties of overall multifactorial native (non-anticoagulated) blood. However, this assumption is wrong and needs to change, not least in particular because relevant in vivo wall shear rates are not included in these tests.A different approach based on physiological (functional) and biological (evolutionary) ideas was proposed by separate groups in the 1970s. The first group was made up of Baumgartner, Sakariassen and colleagues [1–5], and the other of Kovacs and colleagues [6–10]. Both used native blood as samples (ex vivo) and thrombosis was measured at various shear rates. The Baumgartner, Sakariassen and colleagues thrombosis tests are triggered by various thrombogenic surfaces, including human arterial subendothelium, human fibrillar collagen and human tissue factor/phospholipids at wall shear rates varying from 100 to 32,000 s-1. Blood is drawn directly from an antecubital vein over the prothrombotic surface at various controlled-wall shear rates, thus avoiding coagulation and platelet activation before reaching the prothrombotic surface [4]. These tests have been successfully used in academic and biopharma research since the 1970s. They are used in drug discovery and development and in therapeutic procedures, including optimization of antiplatelet agents and anticoagulation at various wall shear rates [3]. In the Global Thrombosis Tests developed by Kovacs and colleagues, thrombus formation is triggered by high shear stress.A thrombus is formed by the interaction between blood and blood vessels under blood flow (Virchow's triad). Yamamoto et al. introduced the helium–neon laser-induced in vivo thrombosis system established by Kovacs et al. Subsequently they began research with shear-induced thrombosis/thrombolysis (fibrinolysis) tests both in animals and humans using native blood (ex vivo). Yamamoto and colleagues have analyzed the matching results obtained by ex vivo and in vivo tests in animal experiments. Ex vivo and in vivo results were closely correlated, although with exception in rodents with severe endothelial dysfunction, recommending a simultaneous endothelial function test (flow-mediated vasodilation test) [11]. No correlation was found between the results obtained by testing diabetics and patients after stroke with a shear-induced thrombosis test using native blood (thrombotic status analyser) and conventional agonist-induced platelet aggregation measurement using anticoagulated blood [12].Yamamoto et al. observed qualitative differences between fruit and vegetable varieties using shear-induced thrombosis/fibrinolysis ex vivo tests and He–Ne laser-induced thrombosis in vivo test. They demonstrated that the antithrombotic activity of fruits and vegetables varies within the same species, that is, there are varieties with antithrombotic activity, those with prothrombotic activity and those with neither effect [13]. In healthy volunteers, antithrombotic status was observed within 2 h of oral administration of an antithrombotic vegetable variety [14]. Further to this, a daily intake of antithrombotic fruit and vegetable varieties over 12 weeks significantly lowered the thrombotic status of humans [15]. Yamamoto and colleagues proposed that the exercise paradox could be avoided by assessing individual thrombotic status with shear-induced thrombosis/fibrinolysis tests [16]. Similar results have been demonstrated by Sakariassen and colleagues in studies focusing on diet supplement, physical exercise and cigarette smoking [3,17,18].The ex vivo Global Thrombosis Test (GTT) is based on the principles of flow chamber techniques, first described by Baumgartner and Sakariassen, and their colleagues [1,4]. The usefulness of the GTT has already been proven in healthy volunteers [19–21] and in patients [22–25]. Tests using native blood have not been widely used in clinical settings. GTT was first described in 2003 [9] and the first clinical experience with the technique was published by Nishida et al. [26]. GTT has been cited in several review articles [27–29]. The slow increase in the number of publications on GTT might be due to the fast coagulation of non-anticoagulated blood, and hence the concern that tests using native blood might not be suitable to assess thrombotic status in clinical settings. Coagulation is, however, slower under the flow conditions in GTT. Previous studies by Baumgartner, Sakariassen, Kovacs and respective colleagues successfully measuring thrombus formation and resolution provided evidence that the use of native blood under flow conditions is superior to studies using anticoagulated blood. As the variability of test results depends on the technique used to obtain the blood sample, there was concern of high variation regarding test results obtained by GTT. However, reproducibility of the GTT has been tested and published [30]. If blood sampling is performed by a trained operator, the reproducibility of both thrombotic (OT) and thrombolytic activity (LT) is good (the intra-assay coefficient of variation of OT was 10% and 6% for LT; the inter-assay coefficient of variation of OT was 8% and 9% for LT), which allows for credible statistical analysis of the findings. The point-of-care GTT has proven to be sensitive in detecting small differences in thrombotic and fibrinolytic activities. GTT surpassed the sensitivity of routine coagulation tests such as the prothrombin time test and the activated partial thromboplastin time test, especially in monitoring oral anticoagulants [31]. GTT detected hyper thrombotic status after overwork, which could not be detected by conventional coagulation tests [32]. Shear-dependent thrombosis tests, including the Baumgartner, Sakariassen and colleagues’ shear dependent tests and the point-of-care ex vivo Global Thrombosis Test, could be useful in assessing thrombotic status and bleeding of patients, in developing antithrombotic drugs and diets and in proposing antithrombotic programs utlilizing physical exercise for the prevention of thrombotic disorders.Financial & competing interests disclosureThe authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.Author contributionsThese authors contributed equally to this manuscript.Open accessThis work is licensed under the Creative Commons Attribution 4.0 License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/References1 Baumgartner HR, Turitto VT, Weiss HJ. Effect of shear rate on platelet interaction with subendothelium in citrated and native blood. II. Relationships among platelet adhesion, thrombus dimensions, and fibrin formation. J. Lab. Clin. Med. 95(2), 208–221 (1980).Medline, CAS, Google Scholar2 Sakariassen KS, Aarts PA, de Groot PG, Houdijk WP, Sixma JJ. A perfusion chamber developed to investigate platelet interaction in flowing blood with human vessel wall cells, their extracellular matrix and purified components. J. Lab. 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Thrombolysis 45(2), 222–224 (2017).Crossref, Google ScholarFiguresReferencesRelatedDetailsCited ByA point-of-care global thrombosis test measuring occlusion time and endogenous lysis time may indicate thrombotic statusJunichiro Yamamoto, Nobutaka Inoue, Kazunori Otsui, Hideo Ikarugi, Muneshige Shimizu, Shinji Yamamoto, Masahiro Murakami, Yoshinobu Ijiri & Kjell S Sakariassen2 July 2019 | Future Science OA, Vol. 5, No. 6 Vol. 5, No. 2 Follow us on social media for the latest updates Metrics History Received 30 September 2018 Accepted 16 October 2018 Published online 19 November 2018 Published in print February 2019 Information© 2018 Junichiro YamamotoKeywordsdietexercisefibrinolysisnative bloodplatelet functionshear rate-dependent thrombosisthrombolysisthrombusFinancial & competing interests disclosureThe authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.Author contributionsThese authors contributed equally to this manuscript.Open accessThis work is licensed under the Creative Commons Attribution 4.0 License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/PDF download

Prevention of thrombotic disorders has priority over treatment. There are only two pathologically relevant tests which are suitable for measuring the overall thrombotic status both in experimental conditions and in humans. The Global Thrombosis Test (GTT) and the Global Parallel-Plate Thrombosis Test can detect the pathologically relevant global thrombotic status. These tests have been successfully used for monitoring the effect of antithrombotic drugs and for developing novel antithrombotic agents. By using GTT, varieties of fruits, vegetables, and regular physical exercise have been tested for the effect on global thrombotic status. This review discusses the published evidence for the benefit of diet of selected fruit and vegetable varieties and doing regular physical exercise on improving thrombotic status. Future clinical trials monitored by GTT or Global Parallel-Plate Thrombosis Test could decide on the effectiveness of an experimentally proven antithrombotic diet with regular physical exercise in the prevention of thrombotic diseases.

A study was conducted to investigate the relationship between sprinting ability and agility using a longitudinal design. The subjects were 70 junior athletes (39 boys and 31 girls) aged 9 to 12 years participating in a Talent Identification and Development program. Physical constitution (height and body weight), 20-m sprint test time, rebound jump index, and ability to change direction (agility test and T-run test) were measured periodically for 3 years. The main results were as follows: 1. Physical constitution, sprinting ability, rebound jump index, and ability to change direction increased significantly along with development in both boys and girls. 2. Performances in the 20-m sprint, agility test, and T-run test in boys were significantly better than in girls, but no significant sex difference was observed in the rebound jump index. 3. Single correlation analysis demonstrated significant correlations between all of the tests. 4. Partial correlation analysis (using age in months as the control variable) revealed significant relationships between the agility test and T-run test in boys and girls, but correlations between 20-m sprint ability and the agility test or T-run test were not significant. 5. There was little difference in the rebound jump index between boys and girls, and it was suggested that ability to change running direction was less affected by simple sprinting or strength-shortening cycle abilities during the developmental period.

While exercise is widely believed to prevent atherothrombotic diseases, it occasionally causes sudden death. This exercise paradox may be due to the inadequate testing of the thrombotic and thrombolytic status. A recently developed shear-induced thrombosis/endogenous fibrinolysis test performed with non-anticoagulated blood samples allows the assessment of the thrombotic state of an individual both at rest and after exercise. This sensitive and physiologically relevant test may help to solve the aforementioned exercise paradox.

While exercise is widely believed to prevent atherothrombotic diseases, it occasionally causes sudden death. This exercise paradox may be due to the inadequate testing of the thrombotic and thrombolytic status. A recently developed shear-induced thrombosis/endogenous fibrinolysis test performed with non-anticoagulated blood samples allows the assessment of the thrombotic state of an individual both at rest and after exercise. This sensitive and physiologically relevant test may help to solve the aforementioned exercise paradox.

Aims] A number of studies have identified abdominal muscle thickness as a significant risk factor for metabolic syndrome (MetS).However, in the absence of longitudinal studies, the causal relationship between abdominal thickness and MetS risk factors has not been elucidated.In this study, we examined the association between abdominal muscle thickness and changes in MetS risk factors.[Methods] We studied 36 patients receiving hospital outpatient care for obesity and MetS.Measurements were taken on two occasions, with an intervening interval of 6 ± 1 months.Data for the following variables associated with the risk of obesity and MetS.Statistical analysis was performed with partial correlation analysis, including age, sex, and current medication, as control variables.[Results] The individual and the combined effects of the control variables were evaluated.However, no significant correlation between abdominal muscle thickness and changes in MetS risk factors was established, after controlling for all combinations of age, sex, and current medication.The abdomen has relatively limited skeletal muscle mass compared to the legs; therefore, variations in muscle mass may have little effect on MetS risk factors.[Conclusions] Although abdominal muscle thickness is a useful indicator for early detection of sarcopenia in obese individuals, our findings suggest that there is a limited association between abdominal wall thickness and MetS risk factors.

INTRODUCTION:Prevention of arterial thrombotic diseases has high priority in developed countries. Taurine (2-aminomethylsulfonic acid), which is rich in sea foods, showed antithrombotic effect in animal models of thrombosis. The present study aimed to investigate such effect in healthy human volunteers. METHODS AND RESULTS:In 101 healthy Japanese people the overall thrombotic status was accessed from non-anticoagulated blood sample by the Global Thrombosis Test (GTT). There was no significant correlation between taurine concentration in urine samples and GTT-Occlusion Times (OT; mainly reactivity of platelets). In contrast, a significant inverse correlation was demonstrated between urine taurine concentrations and GTT-Lysis Times (LT; showing spontaneous thrombolytic activity). CONCLUSIONS:Our findings suggest that taurine enhances endogenous thrombolytic activity which could be a mechanism of the earlier observed cardioprotective and antithrombotic effect.

Objective To determine and compare thrombotic and endogenous thrombolytic status in Japanese and Western populations. Background Incidence of coronary heart disease (CHD) and AMI in Japan remains lower than in Western countries. Primary genetic effects are unlikely, given the increased CHD in Japanese migrants. For men, cholesterol and blood pressure have been similar in Japan and the U.S. Dietary factors are implicated, but how these effect CHD is unclear. We postulated that differences in thrombotic and/or thrombolytic status may contribute. Methods We measured thrombotic and thrombolytic status in 100 healthy Japanese (J) from Japan and 100 healthy Westerners (W) from the U.K. using the Global Thrombosis Test (GTT). The GTT employs non-anticoagulated blood to create platelet-rich thrombi under high shear (occlusion time OT; seconds), and then measures the restart of blood flow, due to spontaneous thrombolysis (lysis time LT; seconds). Results OT was longer in (J) compared to (W) (545 vs. 364, p<0.0001). LT was longer in (J) than in (W) (1753 vs. 1052, p<0.0001). Distribution of LT in (J) did not conform to a normal population, with markedly impaired thrombolytic status (LT>3000s) in 18%, compared to none of the Westerners (p<0.0001). Conclusions There are marked differences in thrombotic and thrombolytic status, with (J) having less prothrombotic (longer OT) but less favourable endogenous thrombolytic profile (longer LT). This may be important in the aetiology of thrombotic events. Since platelets and thrombolysis were both inhibited in (J) and yet incidence of AMI is lower, OT would seem more important than LT as a determinant of overall thrombotic risk in this population.

2000 年 3 月,日本政府は生活習慣病対策と して「21 世紀における国民健康づくり運動」 (健康日本 21)を定めた.しかし,「平成 16 年 国民健康・栄養調査」により,運動習慣をもつ 者の割合は,男性 30.9%,女性 25.8%であり, 「健康日本 21」の取り組みにもかかわらず,国 民の 3 分の 2 以上が運動習慣を身につけてい ない状況が明らかになった.そして,厚生労働 省の中における健康増進に関する審議会の中で 最も上位である厚生科学審議会地域保健健康増 進栄養部会において,今後の生活習慣病対策 は,「1 に運動,2 に食事,しっかり禁煙,最後 にクスリ」の標語のもと,身体活動・運動施策 について,より一層推進することが答申され た.これらの状況を踏まえて,2006 年 7 月, 厚生労働省の「運動所要量・運動指針の策定検 討会」は,現在の科学的知見に基づき,生活習 慣病を予防するための身体活動量・運動量およ び体力の基準値として,「健康づくりのための 運動基準 2006 ─身体活動・運動・体力─」(運動 基準)を取りまとめた.この運動基準は,こ れまでに蓄積された科学的知見,具体的には身 体活動量,運動量,体力と各種生活習慣病発症 に関する大規模コホート研究の系統的レビュー を行い,一定基準をクリアした文献を収集し, そこから得られた値をもとに作成されている. 具体的には表 1 に示すような検索方法によっ てレビューが行われ,最終的に 84 編の文献に 基づき策定されたことが公表されている.しか し,表 1 に示されている通り,対象とした生 活習慣病は肥満や高血圧症などであり,血栓止 血機能に及ぼす運動の影響について直接的には 扱われていない.運動が生活習慣病対策に重要 な柱であることに疑いはないものの,一方で運 動が冠動脈疾患を誘発することも報告され, “ 運 動 の パ ラ ド ッ ク ス(the paradox of exercise)” あ る い は“ 諸 刃 の 刃(the twoedged sword)”として指摘されている .身体 血栓止血機能に及ぼす運動の影響 鵤 木 秀 夫 ,山本順一郎 2

Coronary allograft vasculopathy is an inflammatory-proliferative process that compromises the long-term success of heart transplantation and currently has no effective prevention and treatment. Lipid nanoparticles, termed LDE can carry chemotherapeutic agents in the circulation and concentrates them in the heart.Twenty-eight rabbits fed a cholesterol-rich diet and submitted to heterotopic heart transplantation were treated with cyclosporine A (10 mg/kg daily) and allocated to four groups of 7 animals treated with intravenous LDE-methotrexate (MTX; 4 mg/kg weekly), with LDE-paclitaxel (PACLI; 4 mg/kg weekly), or with LDE-PACLI (4 mg/kg weekly) and LDE-MTX (4 mg/kg weekly). A control group was treated with only weekly intravenous saline solution. Animals were euthanized 6 weeks later for morphometric, histologic, immunohistochemical, and gene expression analysis of the graft and native hearts.Compared with controls, grafts of rabbits treated with LDE-PACLI showed 50% reduction of coronary stenosis, and in the LDE-MTX and LDE-MTX/PACLI stenosis was approximately 18% less than in control, but this difference was not statistically significant. In the three treatment groups, macrophage infiltration was decreased. In the LDE-MTX group, gene expression of proinflammatory factors tumor necrosis factor-α, monocyte chemoattractant protein 1, interleukin 18, vascular cellular adhesion molecule 1, and matrix metalloproteinase 12 was strongly diminished, whereas expression of antiinflammatory interleukin 10 increased. In the LDE-PACLI and LDE-PACLI/MTX groups, proinflammatory and antiinflammatory gene expressions were not consistently changed by the treatments.LDE-PACLI promoted strong improvement of cardiac allograft vasculopathy, but the decrease in coronary stenosis by LDE-MTX and LDE-MTX/PACLI was not significant. All three treatments decreased macrophage infiltration in the graft. These results may encourage future clinical trials to test this new therapeutic approach to coronary allograft vasculopathy.

INTRODUCTION:We have shown earlier that diacylglycerol (DAG) but not triacylglycerol (TAG) inhibited thrombus formation. The aim of the present study was to investigate the mechanism of this antithrombotic effect of DAG.MATERIALS AND METHODS:Four different diets, the (1) Western-style high-fat diet (HFD) containing 20% lipid and 0.05% cholesterol (w/w), (2) TAG-rich and (3) DAG-rich HFDs containing 20% lipid and 0.05% cholesterol, but all lipid replaced by TAG or DAG oil with very similar fatty acid composition and the (4) Japanese-style low-fat diet (LFD) containing 7% oil but no cholesterol were given to apolipoprotein E and low-density lipoprotein (LDL) receptor double-deficient mice. Atherogenicity was assessed by morphology, mapping the whole aorta and measuring the total area of lipid-stained lesions. Endothelial function was measured by the flow-mediated vasodilation test. Platelet reactivity was assessed from native blood sample by a shear-induced platelet function test (hemostatometry). Serum lipoprotein profile was measured by HPLC.RESULTS:Both the Western-style and the TAG-rich HFDs have accelerated atherosclerosis. In contrast, DAG-rich HFD inhibited the atherosclerotic process to an extent comparable with the Japanese-style LFD. There was no significant difference in platelet and coagulant activity between the studied diet groups. DAG-rich but not the TAG-rich HFD significantly suppressed serum LDL cholesterol level.CONCLUSIONS:The present findings suggest that the mechanism of antithrombotic and anti-atherogenic effect of DAG may involve the protection of the vascular endothelium from injury and lowered serum LDL cholesterol.

The aim of the present study was to investigate the effect of a NO donor (GSNO) and a plasma kallikrein inhibitor (PKSI-527) alone and in combination on global haemostatic status. A new in vitro test was employed which allows the measurement of both platelet function and spontaneous thrombolysis. Sixteen healthy young and 18 elderly volunteers were enrolled in this study. When GSNO (1 mM) or PKSI-527 (20 microM) was added to native human blood, platelet reactivity was significantly inhibited in both age groups. The combination of GSNO and PKSI-527 had additive inhibitory effect on platelets. Addition of either GSNO or PKSI-527 to blood samples did not significantly affect spontaneous thrombolysis, while added together, spontaneous thrombolysis was significantly enhanced. The thrombolysis enhancing effect was more prominent in elderly subjects. Our present findings suggest that the combination of NO donor and plasma kallikrein inhibitor may have clinical antithrombotic potential.

Background: Although incidence of arterial thrombotic events increases with advancing age, the mechanism of such increased risk is poorly understood. There is also a gender difference; coronary heart disease is more common in men than women. The aim of the present study was to investigate the possible contribution from platelet reactivity and spontaneous thrombolysis, determinants of arterial thrombosis, to the increased risk of thromboembolism in aging, but otherwise healthy subjects.Method: One hundred and forty-five normal subjects (61 men, 82 women) of age between 19 and 77 years were grouped into groups of young (< 30 years); middle aged (31-50 years), and older (> 51 years), according to their gender and body mass index (BMI). A new in vitro test [Gorog Thrombosis Test (GTT)] was used to measure in sequence both platelet reactivity [occlusion time (OT)] and spontaneous thrombolysis [lysis time (LT)] from one non-anticoagulated blood sample.Result: OT in all women was 315 +/- 9.2 s (mean +/- S.E.M.), in all men OT was 300 +/- 10.6 s. The mean LT in all women was 2557 +/- 201.5 s, and in all men LT was 2493 +/- 198.6 s. Advanced age did not enhance platelet reactivity (OT), but increased BMI did (P=0.039). Spontaneous thrombolysis (J) was impaired in older men but not in women (difference between young vs. middle age: P=0.019; young vs. older: P=0.0002).Conclusion: Our findings suggest that in men, spontaneous thrombolytic activity is reduced with age, and this may explain the increased frequency and severity of thromboembolic events. Interestingly, in women, spontaneous thrombolytic activity did not change after menopause. (c) 2004 Elsevier Ltd. All rights reserved.

Introduction: Earlier we have demonstrated a prothrombotic state in spontaneously atherogenic rodents kept on Western-style high fat diet. The aim of the present study was to investigate the cellular mechanism of such prothrombotic state.Materials and methods: Two kinds of diets, Western-style high fat diet containing 20% fat (w/w) and 0.05% cholesterol (w/w) and low fat diet containing 7% fat without cholesterol based on AIN93G, were added to diet-sensitive apolipoprotein E and low-density lipoprotein receptor double deficient mate mice for 12 or 18 weeks from 6 weeks of age. Atherosclerosis was assessed by morphometry of the aortic watt or lipid-stained lesions. Endothelial function was measured by flow-mediated vasodilation (FMV) of the femoral artery. Platelet reactivity was measured ex vivo by a shear-induced platelet aggregation test.Results and conclusions: 12 weeks feeding of mice with high fat diet significantly impaired FMV, as compared with mice fed with low fat diet (P < 0.05). In contrast, there was no significant difference in the lipid-stained areas and in the reactivity of platelets between the two groups. 18 weeks feeding with high fat diet significantly impaired FMV (P < 0.05) and enhanced both lipid-stained areas (P < 0.05) and platelet reactivity (P < 0.01). These findings show that in high fat diet-induced prothrombotic state, endothelial dysfunction precedes both the morphologically detectable lesions and the enhancement of platelet reactivity. (c) 2005 Elsevier Ltd. All rights reserved.

A study was conducted to investigate the cardiac autonomic system activity, physical activity, and circadian rhythm of body temperature in children with low waking basal body temperature. Fifty-five boys (aged 9-10 yrs) were evaluated for oral body temperature (OBT) and daily physical activity over a one-week period. Cardiac autonomic nervous system activity was also measured during the same period using a 5-min power spectral analysis of resting heart rate variability. The low temperature group (LOW) was defined as>1 SD below the mean waking OBT (n=6). The 43 individuals with a higher waking OBT score (<1 SD below the mean) served as the control group (C). There were no significant differences in waking time or total sleeping time between LOW and C. However, a low waking OBT, and an insufficient decline in bedtime OBT were observed in LOW. Sympathetic nervous system activity was significantly higher, while parasympathetic nervous system activity was significantly lower in LOW versus C. Daily physical activity was significantly lower in LOW versus C. Our results suggest that reduced parasympathetic nervous system activity and a lower level of physical activity may affect OBT circadian rhythm. Our data also suggest that a low waking OBT may be associated with delayed OBT circadian rhythm.

This is the first laboratory evaluation of a new instrument, designed to test both platelet function and thrombolytic activity from a native blood sample, in vitro. The inventor assumed that the reduction and arrest of blood flow was due to activation, aggregation and stabilized thrombus formation by shear-activated platelets, and that re-establishment of flow was due to thrombolysis. Morphologic and functional studies presented here confirm these mechanisms. In vitro tests provided incontestable evidence for the principal role of platelets in the obstruction of flow (occlusion time) and for thrombolysis as the principal mechanism underlying the restoration of blood flow (lysis time). In addition to aggregation, it is the explosive generation of thrombin by shear-activated platelets that results in the formation of an occlusive haemostatic thrombus. Anticoagulation of blood completely prevented occlusion. Platelet-rich thrombus formation (occlusion time) was dose-dependently inhibited by monoclonal antibody against platelet glycoprotein (GP) Ib (6B4 and 12E4), aurin tricarboxylic acid, monoclonal antibody against platelet GPIIb/IIIa (MA-16N7C2 and abciximab), a synthetic GPIIb/IIIa antagonist (TAK-029), thrombin inhibitor (argatroban), and anti-von Willebrand factor, but not by anti-fibrinogen. Plasminogen activator streptokinase (Varidase) and tissue-type plasminogen activator (Monteplase) dose-dependently enhanced thrombolysis (lysis time) without affecting platelet function (occlusion time). The test is specific for thrombolysis. The plasmin inhibitor tranexamic acid prevented plasminogen activator-induced thrombolysis, while inhibition of clot retraction by cytochalasin B did not affect the lysis time. This rapid and sensitive global test of platelet function and thrombolytic activity could be of great value both in research and in clinical practice.