Differences in serum and plasma levels of microRNAs and their time-course changes after blood collection.

Background: Serum and plasma are used for measurements of microRNAs (miRNAs) as biomarkers of various diseases. However, no consistent findings have been obtained regarding differences in serum and plasma levels of miRNAs. The purpose of this study was to clarify differences in serum and plasma levels of total miRNAs and their time-course changes after blood collection. Methods: Venous blood was collected from healthy men, and samples were prepared at the time points of 0, 15, 30, 60 and 180 min after blood collection for plasma and after clot formation for serum. Levels of total miRNAs were analyzed by the hybridization method using the 3D-Gene miRNA Oligo chip. Results: About one third of 2632 miRNAs tested showed levels high enough for comparison of serum and plasma levels and for investigation of their time-course changes. Levels of 299 miRNAs at time 0 were significantly different in serum and plasma. Levels of representative platelet-derived miRNAs including miR-185-5p, -22-3p and -320b were significantly higher in plasma than in serum, while levels of representative erythrocyte-derived miRNAs including miR-451a, -486-5p and -92a-3p were not significantly different in serum and plasma. Plasma levels of 173 miRNAs and 6 miRNAs showed significant decreasing and increasing tendencies, respectively, while there were no miRNAs in serum that showed significant time-course changes. Conclusion: The results suggest that careful attention should be paid when comparing serum and plasma levels of miRNAs and that plasma samples should be prepared early after blood collection.

Associations of habitual alcohol drinking with HDL cholesterol and D-dimer in patients with peripheral arterial disease.

BACKGROUND: The risk of cardiovascular disease is lower in light-to-moderate alcohol drinkers than in nondrinkers in the general population. However, it remains to be clarified whether these beneficial effects of alcohol are shown in patients with peripheral arterial disease (PAD). METHODS: The subjects were 153 male outpatients with PAD and they were divided by frequency of drinking into nondrinkers, occasional drinkers (1-4 days/wk) and regular drinkers (5-7 days/wk). Relationships of alcohol drinking with variables related to progression of atherosclerosis and cardiovascular risk were investigated. RESULTS: The levels of HDL cholesterol and d-dimer were significantly higher and lower, respectively, in regular drinkers than in nondrinkers, while there were no significant differences in BMI, blood pressure, total cholesterol, LDL cholesterol, triglycerides, hemoglobin A1c, platelet count, fibrinogen, ankle brachial index and intima-media thickness of the carotid artery in non-, occasional and regular drinkers. Odds ratios of regular drinkers vs. nondrinkers for low HDL cholesterol (0.24 [0.08 âˆ¼ 0.70]) and high d-dimer (0.29 [0.14 âˆ¼ 0.61]) were significantly lower than the reference level. CONCLUSIONS: In patients with PAD, habitual alcohol drinking was associated with increase of HDL cholesterol and suppression of blood coagulability. However, progression of atherosclerosis was not different in nondrinkers and drinkers.

Peptides associated with hypertensive disorders of pregnancy as possible biomarkers for severity of lower extremity arterial disease.

BACKGROUND AND AIMS: Seven circulating peptides, consisting of 18-28 amino acids, were identified as possible biomarkers of hypertensive disorders of pregnancy (HDP) in our previous study. However, it is unknown whether these peptides are relevant to cardiovascular disease. The purpose of this study was to clarify the relationships between serum levels of these peptides and leg arterial blood flow in patients with lower extremity arterial disease (LEAD). METHODS: The subjects were 165 outpatients with LEAD. Patients with advanced LEAD (stages 5 and 6 of the Rutherford classification) were not included. Leg arterial blood flow was evaluated by ankle brachial pressure index (ABI) and % decrease in ABI after leg exercise induced by a leg loader or treadmill. Concentrations of the seven peptides with m/z 2081 (P-2081), 2091 (P-2091), 2127 (P-2127), 2209 (P-2209), 2378 (P-2378), 2858 (P-2858) and 3156 (P-3156) were measured simultaneously with a mass spectrometer. RESULTS: P-2081, P-2127 and P-2209 levels showed significant positive correlations with leg arterial blood flow, while P-2091, P-2378 and P-2858 levels showed significant inverse correlations with leg arterial blood flow. There was no significant correlation between P-3156 levels and leg arterial blood flow. The above positive and inverse associations between peptide levels and leg arterial blood flow were also found in logistic regression analysis using tertile groups divided by the concentrations of each peptide. CONCLUSIONS: Serum levels of six HDP-related peptides (P-2081, P-2091, P-2127, P-2209, P-2378 and P-2858) were associated with lower extremity arterial blood flow in patients with LEAD, and thus these peptides are possible biomarkers for severity of LEAD.

Hematometabolic Index as a New Discriminator of Cardiometabolic Risk in Middle-Aged Men with Polycythemia and High Leukocyte Count in Peripheral Blood.

Background: Both polycythemia and high leukocyte count are associated with the risk of cardiovascular disease. However, it remains to be determined whether polycythemia and high leukocyte count show synergistic increasing effects on cardiometabolic risk. Methods: Cardiometabolic risk was evaluated by cardiometabolic index (CMI) and metabolic syndrome in a cohort of middle-aged men (n = 11,140) who underwent annual health check-up examinations. The subjects were divided into three tertile groups by hemoglobin concentration or leukocyte count in peripheral blood, and their relations with CMI and metabolic syndrome were investigated. A new index, named hematometabolic index (HMI), was defined as the product of hemoglobin concentration (g/dL)-minus-13.0 and leukocyte count (/µL)-minus-3000. Results: When the subjects were further classified by tertiles for hemoglobin concentration and leukocyte count into nine groups, the odds ratios for high CMI and metabolic syndrome of the group categorized in the highest (third) tertiles for both hemoglobin concentration and leukocyte count versus the group of the lowest (first) tertiles for both of them were highest among the nine groups. In receiver-operating characteristic (ROC) analysis for relationships of HMI with high CMI and metabolic syndrome, areas under the ROC curves (AUCs) were significantly larger than the reference level and tended to be smaller with an increase in age. In subjects from 30 to 39 years of age, the AUC for the relationship between HMI and metabolic syndrome was 0.707 (0.663-0.751) and the cutoff of HMI was 9850. Conclusions: HMI, reflecting hemoglobin concentration and leukocyte count, is thought to be a possible marker for discriminating cardiometabolic risk.

Effects of age on polycythemia, cardiometabolic risk and their associations in middle-aged men.

Background: Polycythemia has been reported to be associated with cardiometabolic risk factors. However, it remains to be determined whether age affects their associations. Methods: The subjects were 11,261 men at ages of 30 ~ 65 years who had received annual health checkup examinations. They were divided by age into four groups of 30 ~ 39, 40 ~ 49, 50 ~ 59 and 60 ~ 65 years. Variables related to polycythemia and cardiometabolic risk and their associations were compared in the different age groups. Results: The prevalences of polycythemia and metabolic syndrome tended to be lower and higher, respectively, with an increase of age. Odds ratios (ORs) of subjects with vs. subjects without polycythemia for high LDL-C/HDL-C ratio and metabolic syndrome were significantly high in the age group of 30 ~ 39 years when compared with the reference level (OR with 95% confidence interval: 3.21 [2.35 ~ 4.37] [high LDL-C/HDL-C ratio] and 3.49 [2.38 ~ 5.12] [metabolic syndrome]) and tended to be lower with an increase of age (60 ~ 65 years, OR with 95% confidence interval: 1.36 [0.63 ~ 2.93] [high LDL-C/HDL-C ratio] and 1.88 [1.16 ~ 3.03] [metabolic syndrome]). Conclusion: The prevalence of polycythemia was higher and its associations with cardiometabolic risk such as high LDL-C/HDL-C ratio and metabolic syndrome were stronger in the youngest group than in the older groups. The results suggest that early detection and correction of polycythemia are more effective than its later correction for prevention of cardiovascular disease in middle-aged men.

Overlooking of Individuals with Cardiometabolic Risk by Evaluation of Obesity Using Waist Circumference and Body Mass Index in Middle-Aged Japanese Women.

Waist circumference is often used for the diagnosis of visceral obesity and metabolic syndrome. In Japan, obesity in women is defined by the government as a waist circumference of ≥90 cm and/or BMI of ≥25 kg/m2. However, there has been a controversy for almost two decades as to whether waist circumference and its above-optimal cutoff are appropriate for the diagnosis of obesity in health checkups. Instead of waist circumference, the waist-to-height ratio has been recommended for the diagnosis of visceral obesity. In this study, the relationships between the waist-to-height ratio and cardiometabolic risk factors, including diabetes, hypertension and dyslipidemia, were investigated in middle-aged Japanese women (35~60 years) who were diagnosed as not having obesity according to the above Japanese criteria of obesity. The percentage of subjects showing normal waist circumference and normal BMI was 78.2%, and about one-fifth of those subjects (16.6% of the overall subjects) showed a high waist-to-height ratio. In subjects with normal waist circumference and normal BMI, odds ratios of high vs. not high waist-to-height ratio for diabetes, hypertension and dyslipidemia were significantly higher than the reference level. A considerable proportion of women who have a high cardiometabolic risk might be overlooked at annual lifestyle health checkups in Japan.

Experimental conditions and protein markers for redifferentiation of human coronary artery smooth muscle cells.

A phenotype switch from contractile type to proliferative type of arterial smooth muscle cells is known as dedifferentiation, but to the best of our knowledge, little is known about redifferentiation of coronary artery smooth muscle cells. The purpose of the present study was to determine in vitro culture conditions for inducing redifferentiation of coronary artery smooth muscle cells. In addition, the present study aimed to determine protein markers for detection of redifferentiated arterial smooth muscle cells. Human coronary artery smooth muscle cells (HCASMCs) were cultured in the presence or absence of growth factors, including epidermal growth factor, fibroblast growth factor-B and insulin. Protein expression and migration activity of HCASMCs were evaluated using western blotting and migration assay, respectively. In HCASMCs 5 days after 100% confluency, expression levels of α-smooth muscle actin (α-SMA), calponin, caldesmon and SM22α were significantly increased, while expression levels of proliferation cell nuclear antigen (PCNA) and S100A4 and migration activity were significantly decreased, compared with the corresponding levels just after reaching 100% confluency, indicating that redifferentiation occurred. Redifferentiation was also induced in a low-density culture of HCASMCs in the medium without growth factors. When the culture medium for confluent cells was replaced daily with fresh medium, the expression levels of α-SMA, caldesmon, SM22α, PCNA and S100A4 and migration activity were not significantly different but the calponin expression was significantly increased compared with the levels in dedifferentiated cells just after reaching 100% confluency. Thus, redifferentiation was induced in HCASMCs by deprivation of growth factors from culture medium. The results suggested that α-SMA, caldesmon and SM22α, but not calponin, are markers of redifferentiation of HCASMCs.

Associations between leukocyte count and lipid-related indices: Effect of age and confounding by habits of smoking and alcohol drinking.

Leukocyte count in peripheral blood is an acute-phase reactant and is associated with the risk of atherosclerotic diseases. Blood lipid profile, a major risk factor of cardiovascular disease, is known to be associated with leukocyte count, but it remains to be determined how this association is affected by other factors including lifestyle and age. The subjects were 11261 Japanese middle-aged men (30~65 years old) who had received health checkup examinations. The relationships of leukocyte count with lipid-related indices (ratio of LDL cholesterol to HDL cholesterol [LDL-C/HDL-C], ratio of triglycerides to HDL cholesterol [TG/HDL-C] and cardiometabolic index [CMI]) were investigated. Leukocyte count, LDL-C/HDL-C, TG/HDL-C and CMI were significantly higher in smokers than in nonsmokers, while leukocyte count and LDL-C/HDL-C were significantly lower in regular drinkers than in nondrinkers. Both in overall subjects and subjects without habits of smoking and drinking, LDL-C/HDL-C, TG/HDL-C and CMI were significantly higher in the 2nd and 3rd tertiles for leukocyte count than in the 1st tertile and tended to be higher with an increase of the tertile. Odds ratios for high TG/HDL-C and high CMI of the 3rd vs. 1st tertiles for leukocyte count tended to be lower with an increase of age, and odds ratios for high TG/HDL-C and high CMI of the interaction term, consisting of age (60~65 vs. 30~39 years) and tertile (3rd vs. 1st tertiles for leukocyte count), were significantly lower than the reference level. In conclusion, leukocyte count is associated with lipid-related indices, and the associations are independent of smoking and alcohol drinking and tend to be weaker with an increase of age in Japanese middle-aged men.

Augmentation by resveratrol of the inhibitory effect of ethanol on platelet aggregation.

Ethanol and resveratrol have been shown to inhibit platelet aggregation. The aim of this study was to determine whether resveratrol has an additional effect on ethanol-induced inhibition of platelet aggregation. Ca2+ entry and subsequent aggregation of human platelets were measured by the fluorescence method and light transmittance method, respectively. Thromboxane B2 concentrations in media containing platelets were measured by using the enzyme-linked immunosorbent assay. Platelet aggregation induced by thrombin (0.025 U/ml) was significantly inhibited by preincubation of platelets with ethanol (0.5%). Preincubation with resveratrol (3.125 µM), which did not affect thrombin-induced platelet aggregation, significantly augmented the inhibitory effect of ethanol on platelet aggregation. Similar synergic effects of ethanol and resveratrol were found on aggregatory responses to collagen (2 µg/ml) and arachidonic acid (0.25 mM). On the other hand, the thrombin-induced increase in intracellular Ca2+ concentration ([Ca2+]i) was not affected by ethanol alone, resveratrol alone or both ethanol and resveratrol together. In nominally Ca2+-free medium, arachidonic acid (0.75 mM) caused a potent platelet aggregation, which was not affected by the presence of ethanol alone, resveratrol alone, or both of them together. Thromboxane B2 formation induced by thrombin was significantly inhibited by ethanol (0.5%) alone and resveratrol (3.125 µM) alone, and these inhibitory effects were significantly augmented in the presence of both ethanol and resveratrol together. Resveratrol shows an additive effect on ethanol-induced inhibition of platelet aggregation. This effect by resveratrol is partly explained by its inhibitory action on thromboxane A2 production in platelets. In addition, both ethanol and resveratrol attenuate platelet aggregation through acting on the Ca2+-dependent intra-platelet pathway after an increase in [Ca2+]i induced by thrombin.

Attenuation by habitual alcohol drinking of the associations of obesity with blood pressure and pulse pressure.

BACKGROUND: Obesity and alcohol drinking are known to be risk factors for hypertension. However, it remains to be determined whether alcohol affects the relationships of obesity with blood pressure and pulse pressure. METHODS: The participants in this study were 29 171 men who had received annual health checkups. Relationships of BMI with blood pressure and pulse pressure were investigated in four subject groups divided by average daily alcohol consumption (grams of ethanol/day), non-, light (<22), moderate (≥22 and <44) and heavy (≥44) drinkers. RESULTS: BMI was significantly correlated with SBP and DBP levels both in nondrinkers and drinkers. The strength of the correlations was significantly weaker in drinkers than in nondrinkers. Odds ratios for hypertension in subjects with vs. subjects without obesity tended to be lower with an increase in alcohol intake (odds ratios with 95% confidence intervals: 4.09 (3.69-4.52) in nondrinkers; 3.11 (2.62-3.68) in light drinkers; 2.87 (2.61-3.16) in moderate drinkers; 2.81 (2.49-3.18) in heavy drinkers). Pulse pressure was weakly but significantly associated with BMI and obesity, and these associations were significantly weaker in heavy drinkers than in nondrinkers. There were significant odds ratios for hypertension and high pulse pressure of the interaction term between obesity and alcohol drinking. CONCLUSION: The associations of BMI with blood pressure and pulse pressure and the associations of obesity with hypertension and high pulse pressure were weaker in drinkers than in nondrinkers. Thus, alcohol drinking attenuates the associations of obesity with hypertension and high pulse pressure.

Paradox of the Relationship between Cardio-Ankle Vascular Index and Ankle-Brachial Index in Patients with Lower Extremity Artery Disease.

Objectives: Measurements of ankle-brachial index (ABI) and toe-brachial index (TBI) are standard examinations for evaluating arterial blood flow in lower extremities and diagnosing lower extremity artery disease (LEAD). It remains to be clarified whether cardio-ankle vascular index (CAVI), a blood pressure-independent parameter of arterial stiffness, is associated with ABI and TBI in patients with LEAD. Methods: The subjects were 165 outpatients with LEAD. Arterial blood flow in lower extremities was evaluated by using ABI, TBI, and the degree of leg exercise-induced reduction of ABI (%). Results: CAVI showed significant positive correlations with ABI and TBI and showed significant inverse correlations with exercise-induced % decrease in ABI. CAVI was significantly higher in the 3rd tertile groups of ABI and TBI than that in the corresponding 1st tertile groups and was significantly lower in the 3rd tertile group of exercise-induced % decrease in ABI than that in the 1st tertile group. The above relationships remained significant after adjustment for age, body mass index, blood pressure, diabetes history, and habitual smoking. Conclusions: Although CAVI is a general parameter reflecting arterial stiffness, CAVI showed paradoxical associations, namely, positive associations with ABI and TBI and an inverse association with exercise-induced % decrease in ABI in patients with LEAD.

Differences in circulating obesity-related microRNAs in Austrian and Japanese men: A two-country cohort analysis.

Background: The prevalence of obesity is higher in Western countries than in East Asian countries. It remains unknown whether microRNAs (miRNAs) are involved in the pathogenesis of the ethnic difference in obesity. The purpose of this study was to determine whether expression levels of circulating obesity-associated miRNAs are different in Europeans and Asians. Methods: The subjects were middle-aged healthy male Austrians (n = 20, mean age of 49.9 years) and Japanese (n = 20, mean age of 48.7 years). Total miRNAs in serum from each subject were analyzed using the 3D-Gene miRNA Oligo chip. miRNAs that showed significant differences between the Austrian and Japanese groups were uploaded into Ingenuity Pathway Analysis (IPA). Results: Among 16 miRNAs that were revealed to be associated with obesity in previous studies and showed expression levels that were high enough for a reasonable comparison, serum levels of 3 miRNAs displayed significant differences between the Austrian and Japanese groups: miR-125b-1-3p was significantly lower with a fold change of -2.94 and miR-20a-5p and miR-486-5p were significantly higher with fold changes of 1.73 and 2.38, respectively, in Austrians than in Japanese. In IPA including all 392 miRNAs that showed significant differences between Austrians and Japanese, three canonical pathways including leptin signaling in obesity, adipogenesis pathway and white adipose tissue browning pathway were identified as enriched pathways. Conclusions: miRNAs are thought to be involved in the ethnic difference in the prevalence of obesity, which may in part be caused by different expression levels of miR-125b-1-3p, miR-20a-5p and miR-486-5p.

Evidence for Indoxyl Sulfate as an Inducer of Oxidative Stress in Patients With Diabetes.

BACKGROUND/AIM: Indoxyl sulfate is a metabolite of tryptophan and its urinary level reflects the status of bacterial flora in the intestine. Indoxyl sulfate possesses prooxidant properties and is implicated in various diseases including chronic kidney disease and cardiovascular diseases. However, the relation of urinary indoxyl sulfate to oxidative stress is not known. PATIENTS AND METHODS: The association of urinary indoxyl sulfate levels with urinary levels of oxidative stress markers, 15-isoprostane F2t and pteridine derivatives, was investigated in 255 patients with type 2 diabetes. Indoxyl sulfate and pteridine derivatives were measured by using spectrofluorometry. RESULTS: Urinary levels of indoxyl sulfate, pteridines, and 15-isoprostane F2t showed a normal distribution after logarithmic transformation but not before it, and they were thus used for parametric analysis after logarithmic transformation. Urinary indoxyl sulfate levels were significantly correlated (p<0.01) with urinary 15-isoprostane F2t and pteridine levels [Pearson's correlation coefficients: 0.503 (15-isoprostane F2t) and 0.562 (pteridines)]. These associations were also found in multivariable analysis after adjusting for age, sex, insulin therapy for diabetes, body mass index, mean arterial pressure, hemoglobin A1c, estimated glomerular filtration rate, urinary albumin, and histories of smoking and alcohol drinking. CONCLUSION: Urinary indoxyl sulfate levels showed associations with urinary levels of oxidative stress markers, and the associations were independent of age, sex, insulin therapy for diabetes, body mass index, blood pressure, glycemic status, renal function, smoking, and alcohol drinking. Indoxyl sulfate appears to be an important determinant of redox balance in patients with diabetes.

Modification of the ALBI-PLT Score for the Prediction of High-risk Varices.

BACKGROUND/AIM: A new scoring system [albumin-bilirubin-platelet (ALBI-PLT) score] was reported for identifying cirrhotic patients without high-risk varices (HRV), and patients with ALBI grade 1 (≤-2.60) and a platelet count over 150×109/l were shown to have a low risk of having HRV. The present study modified the cut-off values of the variables in the ALBI-PLT score. PATIENTS AND METHODS: Among a total of 338 patients with chronic liver diseases, possible cut-off values of the ALBI score and the platelet count were determined by analyzing the first-half group (training cohort: N=169) with the receiver operating characteristic (ROC) method. The utility of the determined values was evaluated in the second-half group (validation cohort: N=169) and total cohort (N=338). In addition, the utility of the modified cut-off values was evaluated in patients with compensated cirrhosis (cirrhotic cohort: N=87). RESULTS: Possible cut-off values of the ALBI score and platelet count were found to be -2.36 and 114×109/l, respectively. In the training cohort, these cut-off values provided a higher ratio of avoiding esophagogastroduodenoscopy than the original ALBI-PLT score (53.3% vs. 25.4%, p<0.01). Consistent results were observed in the validation cohort (28.4% vs. 15.4%, p<0.01), total cohort (40.8% vs. 20.4%, p<0.01), and cirrhotic cohort (32.2% vs. 11.5%, p<0.01). However, the missing ratio of patients with the HRV was not significantly increased in any cohort studied. CONCLUSION: Modification of the ALBI-PLT score may be useful for predicting patients without HRV.

Associations between polycythemia and cardiometabolic risk factors in middle-aged men.

BACKGROUND: Polycythemia increases blood viscosity and causes thrombogenesis in cardiovascular disease. However, relationships between polycythemia and cardiometabolic risk factors in a general population remain to be determined. METHODS: Subjects were middle-aged Japanese men (n = 11261) receiving annual health checkup examinations. Relationships of polycythemia, defined as hemoglobin concentrations of 16.5 g/dl or higher, with cardiometabolic risk factors, including obesity, hypertension, dyslipidemia and diabetes, were investigated. RESULTS: Both in univariable analysis and multivariable analysis with adjustment for age, histories of smoking, alcohol drinking and regular exercise, and medication therapy for hypertension, dyslipidemia or diabetes, waist-to-height ratio, systolic and diastolic blood pressure, triglycerides, LDL cholesterol, cardiometabolic index (CMI) and hemoglobin A1c were significantly higher and HDL cholesterol was significantly lower in the polycythemic group than in the non-polycythemic group. The above associations between polycythemia and cardiovascular risk factors were also found in the sub-analysis for nonsmokers (n = 4937). The odds ratios (ORs) of the polycythemic vs. non-polycythemic groups for high CMI and metabolic syndrome (MS) were significantly high when compared with the reference level in the multivariable analysis (OR with 95% confidence interval: high CMI, 2.18 [1.91 âˆ¼ 2.50]; MS, 2.39 [2.02 âˆ¼ 2.85]). CONCLUSION: Polycythemia showed smoking-independent associations with cardiometabolic risk factors including visceral obesity, hypertension, dyslipidemia, diabetes and metabolic syndrome. Thus, polycythemia is a potential discriminator of cardiovascular risk in the general population.

Data on urinary pteridines as a discriminator of atherosclerotic risk in patients with diabetes.

Presented here are the supplemental data of the research article "Urinary pteridines as a discriminator of atherosclerotic risk in patients with diabetes" [1]. These data provide the first information on variables that affect urinary levels of pteridines (oxidized-form pteridine derivatives) in patients with diabetes mellitus. In linear regression analysis, gender (women vs. men), current history of smoking and urinary albumin showed significant positive correlations with pteridines, while there were significant inverse correlations of pteridines with a history of alcohol drinking and body mass index. The above associations were confirmed by using analysis of covariance and logistic regression analysis. Among the eight variables (age, gender, medication therapy for diabetes, smoking, alcohol drinking, body mass index, hemoglobin A1c and urinary albumin) tested, smoking showed the strongest association with urinary pteridines.

VEGF-Independent Angiogenic Factors: Beyond VEGF/VEGFR2 Signaling.

Tumors induce angiogenesis to acquire oxygen and nutrition from their adjacent microenvironment. Tumor angiogenesis has been believed to be induced primarily by the secretion of vascular endothelial growth factor-A (VEGF-A) from various tumors. VEGF-A binds to VEGF receptor 2 (VEGFR2), resulting in subsequent activation of cellular substances regulating cell proliferation, survival, and angiogenesis. Antiangiogenic therapies targeting the VEGF-A/VEGFR2 axis, including bevacizumab and ramucirumab, humanized monoclonal antibodies against VEGF-A and VEGFR2, respectively, have been proposed as a promising strategy aimed at preventing tumor growth, invasion, and metastasis. Phase III clinical trials using bevacizumab and ramucirumab have shown that not all tumor patients benefit from such antiangiogenic agents, and that some patients who initially benefit subsequently become less responsive to these antibodies, suggesting the possible existence of VEGF-independent angiogenic factors. In this review, we focus on VEGF-independent and VEGFR2-dependent tumor angiogenesis, as well as VEGFR2-independent tumor angiogenesis. Additionally, we discuss VEGF-independent angiogenic factors which have been reported in previous studies. Various molecular targeting drugs are currently being evaluated as potential antitumor therapies. We expect that precision medicine will permit the development of innovative antiangiogenic therapies targeting individual angiogenic factors selected on the basis of the genetic screening of tumors.

Associations of Blood Urate Level with Glycemic Status and Other Cardiometabolic Risk Factors in Middle-Aged Women.

Background: Hyperuricemia is a risk factor of cardiovascular disease. It remains to be elucidated how blood urate level is associated with hyperglycemia in women. Methods: The participants were 4612 middle-aged Japanese female workers. They were divided into four quartile groups by serum urate level, and cardiovascular risk factors were compared in the quartile groups. Results: With an increase of the quartile for urate, the means of waist-to-height ratio, systolic and diastolic blood pressure, log-transformed triglycerides, low-density lipoprotein (LDL) cholesterol, and cardiometabolic index (CMI) tended to be higher and high-density lipoprotein (HDL) cholesterol tended to be lower. Hemoglobin A1c was significantly higher in the 4th quartile for urate than in the 1st quartile, but this difference was not found when body mass index (BMI) was adjusted. The odds ratios versus the 1st quartile for high waist-to-height ratio, hypertension, hypertriglyceridemia, hypo-HDL cholesterolemia, hyper-LDL cholesterolemia, high CMI, and diabetes tended to be higher with an increase of the quartile. The odds ratios of the 4th versus 1st quartiles for these abnormalities except for high waist-to-height ratio and diabetes were significantly higher than the reference level even with adjustment for BMI. Hemoglobin A1c showed a weak but significant positive correlation with urate in analysis with adjustment for BMI. Conclusion: Blood urate was positively associated with adiposity, blood pressure, triglycerides, LDL cholesterol, and glycemic status and was inversely associated with HDL cholesterol in middle-aged women. The associations of urate with blood pressure, blood lipids, and glycemic status remained independent of adiposity, although being confounded by adiposity.

Associations of cardiovascular risk with circulating peptides related to hypertensive disorders of pregnancy.

We previously identified seven peptides in serum that are associated with hypertensive disorders of pregnancy (HDP). However, the significance of these peptides in the general population is unknown. The aim of this study was to clarify the relationships of HDP-associated peptides with hypertension and other cardiovascular risks in adult men. We investigated the relationships of peptide levels with cardiovascular risk factors, including adiposity, blood pressure, blood lipids and glycemic status, in men (mean age: 46.4 years) who were receiving annual health checkups at their workplace. The concentrations of the abovementioned seven peptides in serum were measured simultaneously using a mass spectrometer. Among the seven peptides, only a peptide with m/z 2091 (P-2091) derived from fibrinogen-α showed a significant correlation with diastolic blood pressure (Spearman's rank correlation coefficient [r], -0.446). Another peptide with m/z 2378 (P-2378) originating from complement component 4 showed a significant positive correlation with body mass index (r, 0.273) and a significant inverse correlation with HDL cholesterol (r, -0.336). In addition, a peptide with m/z 3156 (P-3156) derived from an inter-α-trypsin inhibitor showed significant inverse correlations with body mass index (r, -0.258) and triglycerides (r, -0.334). There was no significant correlation of the levels of any of the seven peptides with hemoglobin A1c. Among the seven peptides related to HDP, P-2091, P-2378 and P-3156 were inversely associated with diastolic blood pressure, HDL cholesterol and triglycerides, respectively. Therefore, these peptides are possible biomarkers for discriminating cardiovascular risk in a general population.