Potential Metabolite Biomarkers for Acute Versus Chronic Stage of Ischemic Stroke: A Pilot Study.

BACKGROUND: Metabolome profiling is used to identify biomarkers for acute ischemic stroke (AIS). Previous studies compared metabolite profiles in AIS and healthy controls, which did not account for factors that affect metabolome (genetics, medications). This pilot project evaluates the change in metabolite concentrations between the acute and chronic stage of stroke in the same cohort in order to minimize other factors impact. METHODS: We performed global metabolome profile on serum of 20 and urine of 12 stroke patients in acute (72 hours) and chronic (3-5.2 months) stage and compared relative peak values using Wilcoxon and orthogonal partial least squares discriminant analysis methods. Chronic stage metabolite concentrations were considered baseline. We performed analysis to identify significantly overrepresented pathways using MetaboAnalyst. RESULTS: Three serum metabolites asparagine (P = .045), tyrosine (P = .015), and xylose (P = .003) had significantly higher concentrations in acute stage. Seven out of top 10 serum metabolites ranked by Wilcoxon test P value were related to amino acid (AA) metabolism. Two urine metabolites glycine (P = .03) and acetylcarnitine (P = .05) had significantly different concentrations in the acute stage. Five of the top 10 urine metabolites related to AA metabolism. We identified 6 significant pathways after false discovery rate correction that were upregulated in the acute stage: (1) Aminoacyl-tRNA synthesis, (2) nitrogen, (3) alanine, aspartate, and glutamate, (4) branched-chain AA, (5) arginine and proline, and (6) phenylalanine metabolism. CONCLUSION: Longitudinal study design confirms that AA metabolism heavily involved in the pathophysiology of acute brain ischemia. Prospective longitudinal studies with a higher number of participants are needed to establish useful stroke biomarkers.

The role of blood pressure in risk of ischemic and hemorrhagic stroke in type 1 diabetes.

BACKGROUND: Hypertension is one of the strongest risk factors for stroke in the general population, while systolic blood pressure has been shown to independently increase the risk of stroke in type 1 diabetes. The aim of this study was to elucidate the association between different blood pressure variables and risk of stroke in type 1 diabetes, and to explore potential nonlinearity of this relationship. METHODS: We included 4105 individuals with type 1 diabetes without stroke at baseline, participating in the nationwide Finnish Diabetic Nephropathy Study. Mean age at baseline was 37.4 ± 11.9 years, median duration of diabetes 20.9 (interquartile range 11.5-30.4) years, and 52% were men. Office systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured. Based on these pulse pressure (PP) and mean arterial pressure (MAP) were calculated. Strokes were classified based on medical and autopsy records, as well as neuroimaging. Cox proportional hazard models were performed to study how the different blood pressure variables affected the risk of stroke and its subtypes. RESULTS: During median follow-up time of 11.9 (9.21-13.9) years, 202 (5%) individuals suffered an incident stroke; 145 (72%) were ischemic and 57 (28%) hemorrhagic. SBP, DBP, PP, and MAP all independently increased the risk of any stroke. SBP, PP, and MAP increased the risk of ischemic stroke, while SBP, DBP, and MAP increased the risk of hemorrhagic stroke. SBP was strongly associated with stroke with a hazard ratio of 1.20 (1.11-1.29)/10 mmHg. When variables were modeled using restricted cubic splines, the risk of stroke increased linearly for SBP, MAP, and PP, and non-linearly for DBP. CONCLUSIONS: The different blood pressure variables are all independently associated with increased risk of stroke in individuals with type 1 diabetes. The risk of stroke, ischemic stroke, and hemorrhagic stroke increases linearly at blood pressure levels less than the current recommended treatment guidelines.

Mass spectrometry-based urinary metabolomics for the investigation on the mechanism of action of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. leaves against ischemic stroke in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: As a traditional Chinese medicine, Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. leaves (ESL) can treat ischemic, neurasthenia, and hypertension diseases. However, only few studies have been conducted on the mechanism of action of ESL for ischemic disease treatment. AIM OF THE STUDY: This study aimed to discover the potential biomarkers in the rats caused by ischemic stroke and build a gene-enzyme-biomarker network to explore the mechanism of ESL treatment on ischemic stroke further. MATERIALS AND METHODS: The urinary metabolomics strategy was developed by combining UPLC-Q-TOF/MS with multivariate data analysis. The gene-enzyme-biomarker network was built by Cytoscape 3.6.0 on the basis of the potential biomarkers filtered out via urinary metabolomic analysis. Then, the potential target enzymes of ESL in the treatment of ischemic stroke were selected for further validation analysis via the ELISA kits. RESULTS: A total of 42 biomarkers associated with ischemic stroke have been identified, among which 38 species can be adjusted by ESL, including 5'-methylthioadenosine, prostaglandin A2, l-methionine, aldosterone, 11b-hydroxyprogesterone, prostaglandin E3, dehydroepiandrosterone, taurine, 5-methoxyindoleacetate, and p-cresol glucuronide. These biomarkers were involved in several metabolic pathways, including taurine and hypotaurine, arachidonic acid, cysteine and methionine, steroid hormone biosynthesis, tryptophan, and tyrosine metabolism pathways. The gene-enzyme-biomarker network was built, and three predicted target proteins, including cyclooxygenase-2 (COX-2), monoamine oxidase (MAO), and nitric oxide synthase (NOS), were selected as the potential target enzymes for ESL in ischemic stroke treatment. CONCLUSIONS: All results showed that ESL can play a therapeutic role in treating ischemic stroke through different pathways. This study will provide an overall view of the mechanism underlying the action of ESL against ischemic stroke.

A NMR-Based Metabonomics Approach to Determine Protective Effect of a Combination of Multiple Components Derived from Naodesheng on Ischemic Stroke Rats.

Naodesheng (NDS) is a widely used traditional Chinese medicine (TCM) prescription for the treatment of ischemic stroke. A combination of 10 components is derived from NDS. They are: Notoginsenoside R1, ginsenoside Rg1, ginsenoside b1, ginsenoside Rd, hydroxysafflor yellow A, senkyunolide I, puerarin, daidzein, vitexin, and ferulic acid. This study aimed to investigate the protective effect of the ten-component combination derived from NDS (TCNDS) on ischemic stroke rats with a middle cerebral artery occlusion (MCAO) model by integrating an NMR-based metabonomics approach with biochemical assessment. Our results showed that TCNDS could improve neurobehavioral function, decrease the cerebral infarct area, and ameliorate pathological features in MCAO model rats. In addition, TCNDS was found to decrease plasma lactate dehydrogenase (LDH) and malondialdehyde (MDA) production and increase plasma superoxide dismutase (SOD) production. Furthermore, 1H-NMR metabonomic analysis indicated that TCNDS could regulate the disturbed metabolites in the plasma, urine, and brain tissue of MCAO rats, and the possible mechanisms were involved oxidative stress, energy metabolism, lipid metabolism, amino acid metabolism, and inflammation. Correlation analysis were then performed to further confirm the metabolites involved in oxidative stress. Correlation analysis showed that six plasma metabolites had high correlations with plasma LDH, MDA, and SOD. This study provides evidence that an NMR-based metabonomics approach integrated with biochemical assessment can help to better understand the underlying mechanisms as well as the holistic effect of multiple compounds from TCM.

Gender differences in the association of syndecan-4 with myocardial infarction: The population-based Tromsø Study.

BACKGROUND AND AIMS: Cardiovascular disease is a common cause of morbidity and mortality, with gender differences in pathophysiology. The endothelial glycocalyx maintains vascular integrity, and glycocalyx shedding reflects endothelial dysfunction and early atherosclerosis. Syndecan-1 and -4 are components of the glycocalyx, and increased serum levels indicate glycocalyx damage. We hypothesised that increased serum syndecan-1 and -4 were independently associated with myocardial infarction (MI), ischaemic stroke and all-cause mortality in men and women from a general population. METHODS: Using a case-cohort design, we included 1495 participants from the Tromsø Study 2001-02. Syndecan-1 and -4 were measured in serum. Baseline variables also included age, gender, cardiovascular risk factors and urinary albumin-creatinine ratio (ACR). Hazard ratios were assessed using multivariable Cox regression models. RESULTS: Between baseline in 2001-02 and December 2007 fatal or non-fatal MI was experienced by 328 and ischaemic stroke by 191 subjects, and 423 participants died. Syndecan-4 was independently associated with MI (hazard ratio per 10 ng/mL increase 1.32; 95% confidence interval 1.06-1.63), but not ischaemic stroke and mortality, and the associations were unchanged by adjustment for urinary ACR. Interaction between syndecan-4 and sex was borderline significant, and in gender-specific analysis, syndecan-4 was associated with MI in women only. Syndecan-1 was not associated with any endpoint. CONCLUSIONS: Syndecan-4 was associated with incident MI, and the association was stronger in women than in men. This suggests a link between endothelial glycocalyx shedding and coronary heart disease in women. Use of syndecan-4 as a risk marker in clinical setting needs further investigation.

Urinary cadmium concentration and the risk of ischemic stroke.

OBJECTIVES: To examine the association between urinary cadmium levels and the incidence of ischemic stroke and to explore possible effect modifications. METHODS: A case-cohort study was designed nested in the Reasons for Geographic and Racial Differences in Stroke (REGARDS) study, including 680 adjudicated incident cases of ischemic stroke and 2,540 participants in a randomly selected subcohort. Urinary creatinine-corrected cadmium concentration was measured at baseline. Multivariable-adjusted hazard ratios (HRs) and the corresponding 95% confidence intervals (CIs) were estimated with the Barlow weighting method for the Cox proportional hazards regression model. RESULTS: The median urinary cadmium concentration was 0.42 (interquartile range 0.27-0.68) µg/g creatinine. After adjustment for potential confounders, urinary cadmium was associated with increased incidence of ischemic stroke (quintile 5 vs quintile 1: HR 1.50, 95% CI 1.01-2.22, p for trend = 0.02). The observed association was more pronounced among participants in the lowest serum zinc tertile (tertile 3 vs tertile 1: HR 1.82, 95% CI 1.06-3.11, p for trend = 0.004, p for interaction = 0.05) but was attenuated and became nonsignificant among never smokers (tertile 3 vs tertile 1: never smokers: HR 1.27, 95% CI 0.80-2.03, p for trend = 0.29; ever smokers: HR 1.60, 95% CI 1.06-2.43, p for trend = 0.07, p for interaction = 0.51). CONCLUSIONS: Findings from this study suggest that cadmium exposure may be an independent risk factor for ischemic stroke in the US general population. Never smoking and maintaining a high serum zinc level may ameliorate the potential adverse effects of cadmium exposure.

Arsenic Exposure in Relation to Ischemic Stroke: The Reasons for Geographic and Racial Differences in Stroke Study.

BACKGROUND AND PURPOSE: The purpose of this case-cohort study was to examine urinary arsenic levels in relation to incident ischemic stroke in the United States. METHODS: We performed a case-cohort study nested within the REGARDS (REasons for Geographic and Racial Differences in Stroke) cohort. A subcohort (n=2486) of controls was randomly sampled within region-race-sex strata while all incident ischemic stroke cases from the full REGARDS cohort (n=671) were included. Baseline urinary arsenic was measured by inductively coupled plasma-mass spectrometry. Arsenic species, including urinary inorganic arsenic and its metabolites monomethylarsonic acid and dimethylarsinic acid, were measured in a random subset (n=199). Weighted Cox's proportional hazards models were used to calculate hazard ratios and 95% confidence intervals of ischemic stroke by arsenic and its species. RESULTS: The average follow-up was 6.7 years. Although incident ischemic stroke showed no association with total arsenic or total inorganic arsenic, for each unit higher level of urinary monomethylarsonic acid on a log-scale, after adjustment for potential confounders, ischemic stroke risk increased ≈2-fold (hazard ratio=1.98; 95% confidence interval: 1.12-3.50). Effect modification by age, race, sex, or geographic region was not evident. CONCLUSIONS: A metabolite of arsenic was positively associated with incident ischemic stroke in this case-cohort study of the US general population, a low-to-moderate exposure area. Overall, these findings suggest a potential role for arsenic methylation in the pathogenesis of stroke, having important implications for future cerebrovascular research.

Practical Application of Biogenic Amine Profiles for the Diagnosis of Patients with Ischemic Stroke.

BACKGROUND: Ischemic stroke (IS) is still one of the major issues in medicine. Still, early diagnosis and misdiagnosis remain the main barriers for proper patient treatment and follow-up. Exploring new potential diagnostic biomarkers for IS is relevant to decrease patient morbidity and the occurrence of poststroke diseases. Biomedical analysis could bring new light to the background of IS and-in such a way-propose new bioanalytical tools for the early diagnosis, prognostication, and monitoring of IS. MATERIALS AND METHODS: This research aimed to present a discussion on the employment of biogenic amines (BAs), as well as their precursory amino acids and main metabolites, as a new panel of biomarkers for IS. Preliminary patient data were presented and the patients were described with respect to their clinical history and examination records, as well as scientific data gained from the liquid extraction-capillary electrophoresis determination of BAs in the patients' urine samples. RESULTS: The results showed the potential of BA screening using the developed sample preparation and analysis methods in urine during IS, and this will be further studied on a more numerous group of patients with IS to reveal the usefulness of BAs as a new panel of biomarkers for early IS diagnosis and prognostication. CONCLUSIONS: To our best knowledge, this methodology for the first time has been used for the simultaneous analysis of multiple small molecular biomarkers. In addition, the factors that might influence the determination of BAs in real samples were pointed out.

The impact of acute kidney injury on in-hospital mortality in acute ischemic stroke patients undergoing intravenous thrombolysis.

INTRODUCTION: Acute kidney injury (AKI) increases the risk of death in acute ischemic stroke (AIS) patients. Intravenous thrombolytic therapy (iv. rt-PA) seems to be the most effective treatment for AIS patients. The effects of AKI on iv. rt-PA treated AIS cases is less studied. Our paper addresses this issue. METHODS: 45 consecutive stroke patients treated with iv. rt-PA (median age = 64 years; 29 male) and 59 age and sex matched controls not eligible for iv. rt-PA have been enrolled in our study. Subjects were followed-up until hospital release or death (median follow up time = 12 days). RESULTS: The prevalence of AKI did not differ between iv. rt-PA treated patients and controls (35.5% vs. 33.89%). In both groups, AKI was associated with increased in-hospital mortality: 50.0% vs. 3.4% p<0.0001 (in the rt-PA treated), and 45% vs. 30.7% (in controls). AKI iv. rt-PA treated patients had a significantly higher risk of in hospital mortality as compared to the no-AKI iv. rt-PA treated (HR = 15.2 (95%CI [1.87 to 124.24]; P = 0.011). In a Cox-multivariate model, the presence of AKI after iv. rt-PA remained a significant factor (HR = 8.354; p = 0.041) influencing the in-hospital mortality even after correction for other confounding factors. The independent predictors for AKI were: decreased eGFR baseline and elevated serum levels of uric acid at admission, (the model explained 60.2% of the AKI development). CONCLUSIONS: The risk of AKI was increased in AIS patients. Thrombolysis itself did not increase the risk of AKI. In the iv. rt-PA patients, as compared to non-AKI, those which developed AKI had a higher rate of in-hospital mortality. The baseline eGFR and the serum uric acid at admission were independent predictors for AKI development in the iv. rt-PA treated AIS patients.

Values of vanillylmandelic acid and homovanillic acid in the urine as potential prognostic biomarkers in ischaemic stroke patients.

BACKGROUND: Suitable biomarkers that have prognostic values are one of the key points of interest in ischaemic stroke. Increased sympathetic nervous system activity in ischaemic stroke causes multiple local and systemic effects that can be detrimental to the outcome. The mechanism of action is increased secretion and activity of catecholamines, whose end metabolic products are vanillylmandelic acid and homovanilic acid. Aim of our study was to determine whether these compounds can be used as potential prognostic biomarkers in ischaemic stroke, as a unique insight into the activity of the sympathetic nervous system. METHODS: Urine samples of 96 patients with ischaemic stroke and transitory ischaemic attacks were analysed. Values of vanillylmandelic and homovanillic acids in urine were tested using liquid chromatography on the first and third day post-stroke. Severity of stroke was determined using the NIHSS scale, while functional outcome was determined using the Modified Rankin Scale. RESULTS: Values of vanillylmandelic and homovanillic acids positively correlated with functional outcome of ischaemic stroke. Favorable outcomes correlated with decreased values, on contrary to increased values, which were associated with unfavourable outcomes. CONCLUSION: Determining the values of these compounds in the urine is an easily available prognostic tool for the ischaemic stroke outcome, while also influencing potential therapeutic changes.

Urine-Specific Gravity-Based Hydration Prevents Stroke in Evolution in Patients with Acute Ischemic Stroke.

BACKGROUND: Early neurological deterioration after ischemic stroke (stroke-in-evolution [SIE]) is associated with poorer outcomes. Previous studies have demonstrated a link between hydration status and the development of SIE. In this study, we tested the hypothesis that rehydration therapy, administered on the basis of urine-specific gravity (USG) findings, might reduce the development of SIE. METHODS: We conducted a single-arm prospective study of patients with acute ischemic stroke with historical controls. For the study group, a USG higher than 1.010 was taken as an indication for rehydration. Control group patients were rehydrated without referring to USG. An increase in National Institutes of Health Stroke Scale (NIHSS) score of 4 or higher within 3 days was defined as having SIE. RESULTS: A total of 445 patients were analyzed, 167 in the study group and 278 in the control group. The proportion of patients who developed SIE was numerically, but not significantly, lower in the study group (5.9%; 10 of 167) compared with the control group (11.5%; 32 of 278). Among patients with a USG higher than 1.010 at admission, the SIE rate was significantly reduced in the study group compared with the control group (6.1% versus 16.0%; P = .021), while the rate of SIE was similar in those with a USG of 1.010 or lower at admission. Multivariate logistic regression analysis confirmed that USG-based hydration was an independent factor associated with reducing SIE. CONCLUSIONS: USG might be a convenient and useful method for guiding fluid therapy in patients with acute ischemic stroke. USG-based hydration reduced the incidence of SIE among patients with a USG higher than 1.010 at admission.

Prognostic value of urine dipstick proteinuria on mortality after acute ischemic stroke.

BACKGROUND AND AIMS: Proteinuria is a marker of kidney disease and a strong risk factor for cardiovascular diseases including stroke. This study was aimed at investigating the prognostic value of proteinuria measured by urine dipstick in patients with acute ischemic stroke. METHODS: This post-hoc analysis of a prospective cohort study included 3404 consecutive patients who had been admitted for acute ischemic stroke between November 2005 and June 2013. Proteinuria was defined as a trace or more of protein on a urine dipstick test routinely performed at admission. Date and cause of death until December 31, 2013 were collected. We investigated the association of proteinuria with all-cause mortality, cardiovascular mortality (defined as ICD-10 codes I00-I99), and non-cardiovascular mortality. RESULTS: Proteinuria was present in 12.8% of the 3404 patients. During the mean follow-up period of 3.56 ± 2.22 years, there were 681 cases of all-cause mortality (460 cardiovascular deaths and 221 non-cardiovascular deaths). Multivariate Cox regression analysis showed that the presence of proteinuria was an independent risk factor for all-cause mortality (adjusted hazard ratio [HR] 1.69, 95% confidence interval [CI] 1.40-2.04), cardiovascular mortality (adjusted HR 1.65, 95% CI 1.31-2.08), and non-cardiovascular mortality (adjusted HR 1.59, 95% CI 1.13-2.23). Adding proteinuria to the multivariate Cox models moderately improved the model performance for all-cause mortality (integrated area under curve [95% CI]: from 0.800 [0.784-0.816] to 0.803 [0.788-0.818], p = 0.026). CONCLUSIONS: Proteinuria, which was detected on a urine dipstick test, was a significant predictor of mortality after acute ischemic stroke.

Effect of Urinary Kallidinogenase on Transforming Growth Factor-ß1 and High-Sensitivity C-Reactive Protein Expression in Rat Focal Cerebral Ischemic Injury.

BACKGROUND In this study we investigated the effect of urinary kallidinogenase (UK) on transforming growth factor beta 1 (TGF-ß1) expression in brain tissue. We also explored the neuroprotective mechanism of UK against ischemic injury by measuring serum high-sensitivity C-reactive protein (hs-CRP) level changes after rat cerebral ischemic injury. MATERIAL AND METHODS The rat middle cerebral artery ischemia/reperfusion model was established using the suture method. Sprague-Dawley rats were randomly divided into 3 groups: treatment, Gegen control, and blank control. Each group was subsequently divided into 5 subgroups according to time (6, 12, 24, 48, and 72 h). Rats in the treatment group were administered UK as treatment. TGF-ß1 expression was observed at each time point using SABC and immunohistochemical staining methods to estimate cerebral infarct volume percentage. Serum hs-CRP levels were also measured. RESULTS TGF-ß1 protein expression in ischemic brain tissues of the treatment group significantly increased at each time point (P<0.01) compared with both control groups. Treatment group serum hs-CRP levels significantly decreased at each time point (P<0.05) compared with both control groups. CONCLUSIONS UK exerts a neuroprotective effect by upregulating TGF-ß1 expression and inhibiting excessive inflammatory responses.

Exogenous human urinary kallidinogenase increases cerebral blood flow in patients with acute ischemic stroke.

OBJECTIVE: To study the effects of human urinary kallidinogenase (HUK) treatment on acute cerebral ischemia (ACI) using magnetic resonance perfusion weighted imaging (MRP) methods. METHODS: In a non-randomized controlled clinical trial, 30 patients diagnosed with ACI were enrolled and divided manually into 2 groups. The experimental group, consisting of 18 participants, was treated with HUK (0.15 Perinatal Assessment Unit/day) for 7 consecutive days. The control group was treated with routine medication. The participants underwent MRP examination on the first and fourteenth day after onset. The National Institutes of Health Stroke Scale (NIHSS) score, cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT), and time to peak (TTP) were compared between the groups. RESULTS: After undergoing therapy, the experimental HUK-treated group had lower NIHSS scores than the control group (p<0.05). The CBF improved more in the HUK-treated group than in the control group (p<0.05). Additionally, MTT and TTP were shorter in the HUK-treated group than in the control group (p<0.05). CONCLUSION: Human urinary kallidinogenase improves CBF and ameliorates neurological deficits. Human urinary kallidinogenase is a safe and effective treatment approach for treating patients with ACI.

Metabonomic study of the effects of different acupuncture directions on therapeutic efficacy.

Posterior circulation ischemia (PCI) is a common clinical ischemic cerebrovascular disease that can endanger the lives of patients in severe cases. Our previous research found that needling the Fengchi (GB20) acupoint presents a significant effect on PCI and that different acupuncture directions can exert different effects. To investigate the biological mechanism of acupuncture directions, rapid resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry-based metabonomic techniques are used to analyze the metabolic profiles of urine samples. The urine samples were obtained from 30 healthy control subjects, 60 PCI patients before and after treatment of different acupuncture directions. Six metabolites, including LPE (22:6), estrone, uric acid, vanillylmandelic acid, N-acetyl-l-tyrosine, and 4-hydroxyphenylacetylglutamine were identified as potential biomarkers of acupuncture treatment of PCI. Acupuncture treatment of PCI patients significantly changed the levels of these potential biomarkers. Moreover, different acupuncture directions showed different effects on the contents of these biomarkers. These results strongly support the belief that acupuncture direction performs an important function in acupuncture intervention. The findings provide new insights into the mechanism of acupuncture treatment and reveal that acupuncture manipulation results in various curative.

Small Molecule Copper and Its Relative Metabolites in Serum of Cerebral Ischemic Stroke Patients.

BACKGROUND: Copper is a strong pro-oxidant. The most important pro-oxidative form in serum is small molecule copper (SMC), which is copper that is loosely bound to small molecules, such as amino acids and polypeptides. The association between copper and atherosclerotic diseases has been confirmed, but that between SMC and cerebral ischemic stroke (CIS), one of the most principal manifestations and causes of death of atherosclerotic disease, is not yet clear. METHODS: We recruited 45 CIS patients and 25 age- and gender-matched healthy controls. We detected their serum levels of SMC, total copper, homocysteine (Hcy), and ceruloplasmin (CP), as well as urinary total copper, and analyzed the relationship of SMC with these aforementioned metabolites or compounds in CIS patients. RESULTS: SMC was 4.2 ± .5 µg/L and 2.1 ± .9 µg/L; total copper in sera was 1345.5 ± 308.2 µg/L and 1180.3 ± 134.0 µg/L; and total copper in urine was 27.6 ± 9.3 µg/L and 18.8 ± 8.1 µg/L in patients and controls, respectively (all P < .05). Serum CP activity in CIS patients was 59.92 ± 12.11 U/L versus 37.76 ± 5.71 U/L in controls (P = .0001). The concentration of SMC was positively correlated with CP activity, Hcy concentration in sera, and urinary total copper. CONCLUSION: The serum level of SMC and total copper is remarkably elevated, and SMC positively correlates with Hcy, CP activity, and urinary total copper in CIS patients.

¹H NMR-based metabonomic analysis of the effect of optimized rhubarb aglycone on the plasma and urine metabolic fingerprints of focal cerebral ischemia-reperfusion rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The ischemia cerebrovascular disease is one of leading causes of death and long-term disability in modern society. Rhubarb is one of the common traditional Chinese medicine with many effects, and the main pharmacodynamic ingredients are aloe-emodin, rhein, emodin, chrysophanol and physcion. The five components are also known as rhubarb aglycone. Rhubarb aglycone has been confirmed to play a remarkable curative effect on cerebral ischemia, but the mechanism is not clear. In this study, (1)H NMR-based metabonomics approach has been used to investigate the protective effect of the optimized rhubarb aglycone on rats of cerebral ischemia-reperfusion. MATERIALS AND METHODS: Male Wistar rats were divided into four groups: sham operation group, model group, Nimodipine group and the optimized rhubarb aglycone group. Based on (1)H-NMR spectra of plasma and urine, principal component analyses were performed to identify different metabolic markers and explore the changes of associated biochemical pathways. Behavior research and brain histopathology examinations were also performed. RESULTS: It was showed that the optimized rhubarb aglycone treatment improved neurological deficits, cerebral infarction and neuronal apoptosis. Principal component analysis scores plots demonstrated that the cluster of model rats was separated from those of sham operation group; rats of the optimized rhubarb aglycone group were classified from model group, but the optimized rhubarb aglycone group closed to the sham operation group. Optimized rhubarb aglycone regulated the associated amino acid, energy and lipid metabolisms disturbed in model rats. CONCLUSION: Our results suggested that the optimized rhubarb aglycone had protective effect on rats of cerebral ischemia-reperfusion and explored the metabolic regulation mechanism. This work showed that the NMR-based metabonomics approach might be a promising approach to study mechanisms of traditional Chinese medicines.

Comparison of aspirin response measured by urinary 11-dehydrothromboxane B2 and VerifyNow aspirin assay in patients with ischemic stroke.

BACKGROUND: We looked for the prevalence of aspirin nonresponders, compared the results of 2 tests assessing aspirin responses-measurement of urinary 11-dehydrothromboxane B2 (dTXB2) and VerifyNow Aspirin assay-in patients with ischemic stroke, and examined the relationship of aspirin nonresponse and the outcomes of the patients. METHODS: One hundred one patients with ischemic stroke were prospectively included. Aspirin response was assessed by urinary dTXB2 measurement and VerifyNow Aspirin assay. The Spearman correlation coefficients and kappa statistics were calculated to assess correlation and agreement between the 2 tests. The measured outcome was the occurrence of cardiovascular events and death. RESULTS: Prevalence of aspirin nonresponders was 40% and 6%, if they were measured by urinary dTXB2 and VerifyNow Aspirin assay, respectively. Poor correlation in the results between the 2 tests was found (r = .135, P = .190). The degree of agreement between the 2 tests in relation to resistance status was weak (kappa = .032, P = .590). With a mean follow-up time of 17 months, the outcomes occurred significantly higher in aspirin nonresponders who were diagnosed by urinary dTXB2 measurement as compared with patients with aspirin response (18% versus 2%, odds ratio 8.8, 95% confidence interval 1.18-65.4, P = .037). CONCLUSIONS: Our research confirmed poor correlation and lack of agreement between the 2 tests. Only aspirin nonresponders who were diagnosed by dTXB2 measurement were related to having cardiovascular events and death. Further research is still needed to identify the best method of diagnosis of aspirin nonresponders.

Cannabis, ischemic stroke, and transient ischemic attack: a case-control study.

BACKGROUND AND PURPOSE: There is a temporal relationship between cannabis use and stroke in case series and population-based studies. METHODS: Consecutive stroke patients, aged 18 to 55 years, who had urine screens for cannabis were compared with a cohort of control patients admitted to hospital without cardiovascular or neurological diagnoses. RESULTS: One hundred sixty of 218 (73%) ischemic stroke/transient ischemic attack patients had urine drug screens (100 men; mean [SD] age, 44.8 [8.7] years). Twenty-five (15.6%) patients had positive cannabis drug screens. These patients were more likely to be men (84% versus 59%; χ2: P=0.016) and tobacco smokers (88% versus 28%; χ2: P<0.001). Control urine samples were obtained from 160 patients matched for age, sex, and ethnicity. Thirteen (8.1%) control participants tested positive for cannabis. In a logistic regression analysis adjusted for age, sex, and ethnicity, cannabis use was associated with increased risk of ischemic stroke/transient ischemic attack (odds ratio, 2.30; 95% confidence interval, 1.08-5.08). However after adjusting for tobacco use, an association independent of tobacco could not be confirmed (odds ratio, 1.59; 95% confidence interval, 0.71-3.70). CONCLUSIONS: This study provides evidence of an association between a cannabis lifestyle that includes tobacco and ischemic stroke. Further research is required to clarify whether there is an association between cannabis and stroke independent of tobacco. CLINICAL TRIAL REGISTRATION URL: http://www.anzctr.org.au. Unique identifier: ACTRN12610000198022.

Aspirin nonresponders in patients with ischaemic stroke.

From previous studies, the prevalence of aspirin nonresponders is 5.5-45% in patients with various cardiovascular diseases. Those who have aspirin nonresponders have a greater risk of clinically cardiovascular events. The purpose of the study was to look for the prevalence, associated factors and the outcomes of aspirin nonresponders among patients with ischaemic stroke. Patients with ischaemic stroke who were treated during January 2011-August 2011 were included. Urine 11-dehydro-thromboxane B2 (dTXB2) was measured to determine the response to aspirin in patients. The demographics and vascular risk factors were compared between patients who were classified as aspirin responders or aspirin nonresponders. The outcomes of the study were favourable outcome, cardiovascular events and mortality. There were 182 patients included during the study period: 128 patients with an acute ischaemic stroke and 54 patients with a stable ischaemic stroke. Ninety patients (49.5%) were found to be aspirin nonresponders. Multivariate analysis revealed that stroke presentation (acute stroke) was the only factor associated with aspirin nonresponders [odds ratio (OR) 2.38, 95% confidence interval (CI) 1.193-4.746, P = 0.014]. With a mean follow-up time of 16 months, aspirin nonresponders had a less favourable outcome (54 vs. 83%, OR 0.24; 95% CI 0.11-0.51, P < 0.001), marginally higher cardiovascular events (11 vs. 2%, OR 4.48; 95% CI 0.92-21.37, P = 0.045) and higher mortality (12 vs. 1%, OR 10.52; 95% CI 1.3-85.28, P = 0.007). The prevalence of aspirin nonresponders was rather high in Thai patients with ischaemic stroke. Aspirin nonresponders had a less favourable outcome, higher cardiovascular events and death rate.