Ischemic Control Research Articles

Expression of microRNA-21 in acute ischemic stroke: relationship with inflammatory cytokines, clinical severity, and clinical outcome

Background When the brain is deprived of oxygen and nutrients due to stenosis or arterial rupture, neurons in the affected area suffer irreversible damage and cellular death. MicroRNA has been shown to regulate target genes implicated in arterial hypertension, atherosclerosis, and diabetes mellitus, all of which influence the risk of ischemic stroke through inflammation, oxidative stress, cell proliferation, and apoptosis. The study aims to determine the changes in miRNA expression, namely miRNA-21, between acute ischemic stroke patients and controls and their relationship to proinflammatory cytokines, clinical severity, and outcome. Methods Serum samples from tertiary hospitals and controls were used to evaluate miRNA-21 expression as well as cytokines TNF-α, IL-10, ICAM-1, and CCL5 levels within 7 days of stroke onset. The 30-day clinical severity and outcome were assessed using the National Institute of Health Stroke Scale (NIHSS) and modified Rankin Scale (mRS), respectively. Result A total of 64 acute ischemic stroke patients and 22 age-matched controls were recruited, with median ages of 56 and 55.5 years old, respectively. There were more male subjects than females (35 to 29). A statistically significant difference was observed in miRNA-21 expression between patients and controls (p<0.001). This finding implies that miRNA-21 expression may have a contribution in acute stroke patients. This was followed by an increase in proinflammatory markers TNF-α, IL-10, ICAM-1, and CCL5. However, no association was found between miRNA-21 and any pro-inflammatory cytokine. There was no significant correlation between miRNA-21 or cytokines markers with clinical severity or prognosis. Conclusion Our study demonstrated increased miRNA-21 expression and proinflammatory cytokine expression in acute ischemic stroke patients relative to controls. However, this was not related to clinical severity or clinical outcomes.

Characteristics of two different immune infiltrating pyroptosis subtypes in ischemic stroke

BackgroundIschemic stroke (IS) is a serious health hazard and identified as the second leading cause of mortality around the world. However, the role of pyroptosis in the immune microenvironment regulation in IS is still unclear. Here, our study aims to elucidate the effect of pyroptosis on immune microenvironment in IS. MethodsThe regulation mode of pyroptosis in IS was systematically evaluated, and its effects on immune microenvironment were explored, including infiltration of immune cells, immune response gene sets, and human leukocyte antigen (HLA) gene. The genes and drugs related to pyroptosis phenotype were also identified. An MCAO rat model was constructed, and the mRNA expression levels in the classifier model were validated by qRT-PCR. ResultsThe separator is composed of 11 pyroptosis genes, out of which 10 genes could distinguish between ischemic stroke and control samples. CHMP2A, CHMP4A, and NAIP genes are significantly related to immune infiltrating cells, immune response gene sets, and HLA. However, two different pyroptosis subtypes mediated by 10 pyroptosis genes were identified, which were different in immune cell abundance, HLA genes, and immune response gene sets. Furthermore, 199 genes associated with pyroptosis phenotype was identified along with the analysis of biological functions. ConclusionThese findings reveal the potential mechanism of pyroptosis in the immune microenvironment of IS, indicating that pyroptosis functions as a vital component in the complexity and diversity of the immune microenvironment in patients with IS.

Somatostatin receptors in fibrotic myocardium.

A patient with a neuroendocrine tumor and history of coronary artery disease underwent PET with 68Ga-DOTATATE PET tracer for tumor visualization. Analysis of the scan showed uptake of 68Ga-DOTATATE in the left ventricle corresponding to previous myocardial infarct. 68Ga-DOTATATE binds by somatostatin receptors (SSTR) and it has been proposed that it may be useful for the detection of cardiac inflammatory lesions. We aimed to test whether SSTR could be upregulated in cardiac fibrotic scar. We analyzed SSTR in cardiac samples from patients with end-stage ischemic cardiomyopathy (ICM, n = 8) and control hearts (n = 5). In mature ICM tissue, SSTR1 and SSTR2 expression was unchanged and SSTR5 expression was significantly decreased in ICM samples vs. control. Immunohistochemistry showed increased SSTR1 and SSTR2 in ICM. Areas with SSTR1 or SSTR2 staining were often adjacent to fibrotic areas. The majority of SSTR1 and SSTR2 staining localized in cardiomyocytes in fibrotic scar-rich areas where CD68 macrophage staining was not present. SSTR are occasionally upregulated in cardiac fibrotic areas. When using 68Ga-DOTATATE PET tracer to detect cardiac sarcoidosis or atherosclerotic plaque, the possibility of tracer uptake in fibrotic areas should be considered.

Remote ischaemic preconditioning on gene expression and circulating proteins after subacute laparoscopic cholecystectomy: randomized clinical trial.

Surgical stress may lead to postsurgical hypercoagulability, endothelial dysfunction and systemic inflammation, which can impact on patient recovery. Remote ischaemic preconditioning is a procedure that activates the body's endogenous defences against ischaemia and reperfusion injury. Studies have suggested that remote ischaemic preconditioning has antithrombotic, antioxidative and anti-inflammatory effects. The hypothesis was that remote ischaemic preconditioning reduces surgery-induced systemic stress response. During a 24-month period (2019-2021), adult patients undergoing subacute laparoscopic cholecystectomy due to acute cholecystitis were randomized to remote ischaemic preconditioning or control. Remote ischaemic preconditioning was performed less than 4 h before surgery on the upper arm. It consisted of four cycles of 5 min ischaemia and 5 min reperfusion. The gene expression of 750 genes involved in inflammatory processes, oxidative stress and endothelial function was investigated preoperatively and 2-4 h after surgery in both groups. In addition, changes in 20 inflammation- and vascular trauma-associated proteins were assessed preoperatively, 2-4 h after surgery and 24 h after surgery. A total of 60 patients were randomized. There were no statistically significant differences in gene expression 2-4 h after surgery between the groups (P > 0.05). Remote ischaemic preconditioning did not affect concentrations of circulating proteins up to 24 h after surgery (P > 0.05). The study did not demonstrate any effect of remote ischaemic preconditioning on expression levels of the chosen genes or in circulating immunological cytokines and vascular trauma-associated proteins up to 24 h after subacute laparoscopic cholecystectomy in patients with acute cholecystitis.

Comparison between Novel and Conventional Electrocardiographic Markers of Atrial Cardiomyopathy for identification of Persons at Risk for Ischemic Stroke

Abstract Background Atrial cardiomyopathy is associated with increased risk for future atrial fibrillation (AF) and ischemic stroke. Several ECG-derived parameters have been reported to assess the risk for future ischemic stroke. Purpose To predict risk of future ischemic stroke by comparison of the novel parameter duration of the amplified digital sinus-p-wave (APWD; 80mm/mV; 175mm/sec) vs previously described p-wave indices and clinical risk factors (cerebrovascular disease (CVD), CHADS-VASc-score). Methods P-wave-derived parameters were measured on digital 12-lead-sinus-ECGs (recorded +/- 6 months from the clinical stroke event) using the software "ECG-Precision-Analysis V-1-2023". and clinical CHA2DS2-VASc Score was assessed in 80 patients with ischemic stroke (no history of AF, no oral anticoagulation (OAC)) and compared to 80 control patients (without OAC, AF or ischemic stroke within the next five years). Results Patients with ischemic stroke vs control patients presented a higher CHA2DS2-VASc Score (3.7 vs 3.2, p=0.04), had more frequently heart failure (25% vs 13%, p=0.04) and cerebrovascular atherosclerotic disease with stenosis >40% ((22.5% vs 10%, p=0.032). P-wave indices were as follows in patients who developed ischemic stroke within the next 12 months versus patients without stroke within a follow-up period of five years, respectively: APWD (144 vs 128ms, p<0.0001), non-amplified SPWD (118 vs 116ms, p=0.081), A-PTF-V1 (-4660 vs -3860µV*ms, p=0.005), durations and amplitudes of terminal-p-wave in V1 (A-Duration-V1 (83 vs 79ms, p=0.119), A-Amplitude-V1 (-50µV vs -50µV, p=0.028), and advanced interatrial block (aIAB in 25% vs 3.8% of patients; p<0.0001. The novel ECG-derived parameter (duration of the amplified sinus-P-wave >140ms) was the best predictor for future ischemic stroke (accuracy: 74%, sensitivity and specificity: 61% and 86%, respectively; PPV: 82%; area under ROC: 0.776). Sensitivity and specificity of the SPWD, aPTFV1 and aIAB were significantly lower (SPWD>120ms: sensitivity 35% and specificity 78%, PPV: 61%; aPTFV1 ≤-5000µV*ms, accuracy: 61%, sensitivity 46%% and specificity 75%, PPV: 65%; aIAB: accuracy: 61%, sensitivity 25% and specificity 96%, PPV: 87%). Conclusions Among the currently identified ECG markers of atrial cardiomyopathy, the novel APWD and advanced interatrial block have the best predictive performances for diagnosis of persons at risk for ischemic stroke.Comparison between ECG ParametersPredictive Accuracy of Parameters

Abstract 1153: Adipose-derived Mesenchymal Stem-cell Therapy Improves Arteriogenesis, Hemodynamics, And Walking Performance In A Porcine Model Of Peripheral Artery Disease

Background: There are few effective alternatives to revascularization for peripheral artery disease (PAD). We used our porcine model of PAD to test the hypothesis that adipose derived mesenchymal stem cells (ADMSC) injected around the aortic trifurcation in the pelvis improve ischemic limb hemodynamics and walking performance (WP). Methods: In Ossabaw miniswine (N=13, Age=6 mo) with metabolic syndrome, right (R) hindlimb ischemia was induced with R External Iliac and Superficial Femoral artery coil occlusion (T0) via carotid artery access. All swine had weekly testing of WP on a treadmill. At weeks 0 (T0), 4 (T4), and 8 (T8), ankle brachial indices (ABI) and muscle tissue oximetry (STO 2 ) were measured. At T4 and T8 all swine underwent angiography. At T4, in treatment (Tx) swine (N=4) ADMSC were isolated using stromal vascular fractionation of autologous abdominal wall fat. Using jugular vein access, a catheter with a retractable side-deploying needle was inserted in the R iliac vein and stabilized by a juxtaposed angioplasty balloon. The needle was deployed to access the retroperitoneum and ADMSC were infused around the aortic trifurcation. Results: Ischemic controls (IC) had no differences between T4 and T8 in ABIs, and mean STO 2 differences between R and L in calves and thighs. Tx improved both ABI and mean STO 2 measurements (table 1). IC had no changes in WP between T4 and T8, while Tx showed improvements (table1). On angiography, IC had growth of R Internal iliac artery (RIIA, the main inflow to the ischemic R hindlimb) and shorter appearance time (AT) of RIIA compared to L with no difference between T4 and T8. Tx showed increased growth of RIIA and shorter AT at T8 compared to T4 (table 1). Conclusion: In a clinically-relevant (similar to humans in size, anatomy, physiology and comorbidities) porcine model of PAD we found that ADMSC injection around the aortic trifurcation in the pelvis induces arteriogenesis and improves ischemic limb hemodynamics and walking performance.

Association of Glycated Hemoglobin With Acute Ischemic Stroke in a Tertiary Care Center in a Tribal Region of Jharkhand.

Increased glycated hemoglobin (HbA1c) levels have shown an association with an increased risk of stroke in patients admitted to a tertiary care center in Jharkhand. To find out and estimate the risk of acute ischemic stroke in patients with increased HbA1c levels compared with controls. This observational case-control study was conducted on patients admitted to the department of general medicine at a tertiary care center in Ranchi from June 2021 to November 2022. The patients included in this study were those aged 18 years or older and who were clinically and radiologically diagnosed with acute ischemic stroke. Only patients with a first episode of stroke were included, and patients with hemorrhagic stroke or transient ischemic attack were excluded from this study. An equal number of control participants were also included. Ion exchange high-performance liquid chromatography was used to perform the HbA1c tests. The same method was used to measure HbA1c levels in the controls. All findings were recorded in aMicrosoft Excel sheet (Microsoft Corporation, Redmond, WA), and the data were analyzed using SPSS version 22.0software (IBM Corp., Armonk, NY). After performing a descriptive statistical analysis, the findings were classified over a range of values and described accordingly. For each variable, an independent t-test was performed to compare the cases with the controls. A multivariable logistic regression analysis was used to choose the appropriate potential factors to determine the association in the multivariable analysis. A total of 185 cases and 185 controls were included. The mean age of the cases with ischemic stroke was 63.77 ± 10.312, and that of the controls was 53.18 ± 11.35 (p < 0.01). The mean HbA1c level in the patients of acute ischemic stroke was 6.97 ± 1.84, and that of the controls was 5.99 ± 1.69 (p < 0.01). The mean random blood sugar (RBS) value in the ischemic stroke cases was 170.21 ± 84.16, and that of the controls was 150.03 ± 82.25 (p = 0.02). To compare the factors that were determined to be statistically significant between ischemic stroke cases and controls, a multivariable logistic regression analysis was performed. The HbA1c p-value was 0.01, the odds ratio (OR) was 1.280, and the 95% CI was 1.11-1.48. The other variables apart from HbA1c that were statistically significant between the ischemic stroke cases and the controls were age (p < 0.01, OR: 1.280, 95% CI: 1.06-1.11), hypertension (p = 0.618, OR: 1.130, 95% CI: 0.70-1.83), and high-density lipoprotein (p = 0.055, OR: 0.975, 95% CI: 0.95-1.00). When other cofounders were considered, it was concluded that with a 1% increase in HbA1c, the risk of stroke increases by 28% (OR: 1.28, 95% CI: 1.11-1.48). To compare the variables that were determined to be statistically significant between the control and ischemic stroke case groups, a multivariable logistic regression was used. The area under the receiver operating characteristic curve for HbA1c was 0.773and RBS was 0.600. This study shows that higher HbA1c levels in patients increase the risk of ischemic stroke. This study brings to light the need to screen the population periodically for diabetes by routinely testing for Hba1c in those who are at high risk of diabetes. Stroke risk can be reduced with early management and intervention. This study also concludes that HbA1c is a better predictor for assessing the risk of ischemic stroke than RBS levels.

Abstract TP304: Ischemic Stroke Increases Levels of One-Carbon Enzymes and the Folate Receptor in Male and Female Brain Tissue

Stroke is the second most common cause of death worldwide and predominantly affects individuals over 65 years old. Its prevalence is projected to increase in parallel with the aging global population. Nutrition is a modifiable risk factor for ischemic stroke. Folates, B-vitamins and choline play a central role in one-carbon (1C) metabolism, which is a key metabolic network that integrates nutritional signals with biosynthesis, redox homeostasis, epigenetics, regulation of cell proliferation, and stress resistance. Our research group has previously shown that deficiencies in 1C lead to worsened stroke outcomes using preclinical models. However, the impact of ischemic stroke on 1C enzymes remains unknown. The objective of this study is to investigate whether ischemic stroke contributes to a change in the levels of 1C enzymes after ischemic stroke in male and female patients. Brain tissue sections from ischemic stroke patients and controls were stained, all tissue was co-stained with neuronal nuclei (NeuN) and DAPI (4′,6-diamidino-2-phenylindole). The colocalization of all three markers was evaluated by two individuals who were blinded to the experimental groups. Ischemic stroke increased neuronal levels of the folate receptor and 1C enzymes, methylenetetrahydrofolate reductase (MTHFR), thymidylate synthase (TS) and serine hydroxymethyltransferase (SHMT). In male stroke brain tissue was observed to have increased levels of MTHFR, TS, and SHMT. Female brain tissue had increases in the folate receptor and TS. The results suggest that ischemic stroke leads to increased demand on 1C and that there are some small differences between males and females.

Measurement of optic nerve sheath diameter on computed tomography for the differentiation of transient ischemic attacks.

The objective of this study was to investigate whether the measurement of mean optic nerve sheath diameter in patients with transient ischemic attack could be used to distinguish between control groups, the acute ischemic stroke group, and subgroups within the acute ischemic stroke category. Retrospectively, the mean optic nerve sheath diameters of patients aged 18 years and older belonging to control, transient ischemic attack, acute ischemic stroke, and subgroups within the acute ischemic stroke category were measured with initial computed tomography conducted in the emergency department. Out of the 773 patients included in the study, 318 (41.1%) were in the control group, 77 (10%) had transient ischemic attack, and 378 (49%) were categorized as stroke patients. The average mean optic nerve sheath diameter was significantly higher in both the stroke and transient ischemic attack groups compared with the control group (p<0.001 for both comparisons). Furthermore, the mean optic nerve sheath diameter in the stroke subgroups was significantly higher than in both the transient ischemic attack and control groups (p<0.001 for all comparisons). In transient ischemic attack patients, the mean optic nerve sheath diameter showed a significant ability to predict transient ischemic attack (AUC=0.913, p<0.001), with a calculated optimal cutoff value of 4.72, sensitivity of 94.8%, and specificity of 73.9%. The mean optic nerve sheath diameter of patients in the transient ischemic attack group was lower compared with those in the stroke subgroups but higher compared with the control group.

Association of the PDE4D gene variant with selected markers in individuals with ischaemic heart disease: a pilot study.

The aim of the study was to evaluate the variant (rs2910829) of the PDE4D gene in relation to its influence on biochemical, anthropometric and physiological parameters in patients with coronary artery disease and healthy subjects of the Eastern Slovak population. The male group consisted of 72 individuals and the female group consisted of 132 individuals. On the basis of clinical screening the subjects were divided into two groups - with ischaemic heart disease and control group. Genomic DNA was isolated from peripheral blood using a commercial NucleoSpin® Blood Machenery-Nagel kit. Molecular genetic analysis of the polymorphism under study was performed using the StepOne™ Real-Time PCR System instrument. The lipid profile markers TC, HDL, LDL, TG were measured by Cobas Integra 400 plus biochemical analyser, and systolic and diastolic blood pressure using a digital blood pressure monitor. Among anthropometric parameters, body height and weight, waist and hip circumference were measured and BMI and WHR indices were calculated. A statistically significant (p = 0.018) possible association between the mutant T allele and ischaemic heart disease was found in men. In women, we found a statistically significant difference in the systolic (p = 0.013) and diastolic blood parameters (p = 0.005) in the CC genotype. In the group of women, we found statistically significant differences in all observed anthropometric parameters and in LDL and TC markers. In the group of men divided on the basis of BMI, statistical significance was found in systolic blood pressure (p = 0.028). In the group of women with ischaemic heart disease, we found a negative correlation between BMI and HDL. The study contributes to new findings of the representation of genotypes and alleles of the rs2910829 PDE4D gene polymorphism in the Slovak population. This is a pilot study. Interactions between genotype and observed anthropometric, physiological and biochemical markers were confirmed.

Circulating microRNAs as biomarkers of Chagas cardiomyopathy.

Chagas cardiomyopathy (CHCM) is the most important clinical manifestation of Chagas disease. The analysis of cardiac miRNAs may contribute to predicting the progression to CHCM in Chagas indeterminate phase and/or to the differential diagnosis for cardiomyopathy. We carried out a case-control study to identify circulating miRNAs associated with CHCM. We assigned 104 participants to four groups: healthy controls (HC), Chagas non-cardiomyopathy controls, CHCM cases, and ischemic cardiomyopathy controls. We performed a clinical, echocardiographic, and laboratory evaluation and profiled circulating miRNA in the serum samples. Differences between groups were observed in clinical variables and in the analysis of miRNAs. Compared to HC, CHCM participants had 4 over-expressed and 6 under-expressed miRNAs; miR-95-3p and miR-130b-3p were upregulated in CHCM compared with controls, Chagas non-cardiomyopathy and ischemic cardiomyopathy participants, suggesting that might be a hallmark of CHCM. Analysis of gene targets associated with cardiac injury yielded results of genes involved in arrhythmia generation, cardiomegaly, and hypertrophy. Our data suggest that the expression of circulating miRNAs identified by deep sequencing in CHCM could be associated with different cardiac phenotypes in CHCM subjects, compared with Chagas non-CHCM, ischemic cardiomyopathy controls, and healthy controls.

Machine Perfusion Deters Ischemia-Related Derangement of a Rodent Free Flap: Development of a Model

IntroductionMachine perfusion can enable isolated support of composite tissues, such as free flaps. The goal of perfusion in this setting is to preserve tissues prior to transplantation or provide transient support at the wound bed. This study aimed to establish a rodent model of machine perfusion in a fasciocutaneous-free flap to serve as an affordable testbed and determine the potential of the developed support protocol to deter ischemia-related metabolic derangement. MethodsRat epigastric-free flaps were harvested and transferred to a closed circuit that provides circulatory and respiratory support. Whole rat blood was recirculated for 8 h, while adjusting the flow rate to maintain arterial-like perfusion pressures. Blood samples were collected during support. Extracellular tissue lactate and glucose levels were characterized with a microdialysis probe and compared with warm ischemic, cold ischemic, and anastomosed-free flap controls. ResultsMaintenance of physiologic arterial pressures (85-100 mmHg) resulted in average pump flow rates of 360–430 μL/min. Blood-based measurements showed maintained glucose and oxygen consumption throughout machine perfusion. Average normalized lactate to glucose ratio for the perfused flaps was 5-32-fold lower than that for the warm ischemic flap controls during hours 2-8 (P < 0.05). ConclusionsWe developed a rat model of ex vivo machine perfusion of a fasciocutaneous-free flap with maintained stable flow and tissue metabolic activity for 8 h. This model can be used to assess critical elements of support in this setting as well as explore other novel therapies and technologies to improve free tissue transfer.

Diesel exhaust particles exposure exacerbates pro-thrombogenic plasma features ex-vivo after cerebral ischemia and accelerates tPA-induced clot-lysis in hypertensive subjects

The combustion of fossil fuels, mainly by diesel engines, generates Diesel Exhaust Particles (DEP) which are the main source of Particulate Matter (PM), a major air pollutant in urban areas. These particles are a risk factor for stroke with 5.6% of cases attributed to PM exposure. Our aim was to evaluate the effect of DEP exposure on clot formation and lysis in the context of stroke. An ex-vivo clot formation and lysis turbidimetric assay has been conducted in human and mouse plasma samples from ischemic stroke or control subjects exposed to DEP or control conditions. Experimental DEP exposure was achieved by nasal instillation in mice, or by ex-vivo exposure in human plasma. Results show consistent pro-thrombogenic features in plasma after human ischemic stroke and mouse cerebral ischemia (distal MCAo), boosted by the presence of DEP. Otherwise, thrombolysis times were increased after ischemia in chronically exposed mice but not in the DEP exposed group. Finally, subjects living in areas with high PM levels presented accelerated thrombolysis compared to those living in low polluted areas. Overall, our results point at a disbalance of the thrombogenic/lytic system in presence of DEP which could impact on ischemic stroke onset, clot size and thrombolytic treatment.

Simultaneous Pericytes and M2 Microglia Transplantation Improve Cognitive Function in Mice Model of mPFC Ischemia.

Cerebral ischemia is one of the major problems threatening global health. Many of the cerebral ischemia survivors would suffer from the physical and cognitive disabilities for their whole lifetime. Cell based-therapies have been introduced as a therapeutic approach for alleviating ischemia-enforced limitations. Photothrombotic stroke model was applied on the left medial prefrontal cortex (mPFC) of adult male BALB/c mice. Then, pericytes isolated from brain microvessels of adult male BALB/c mice, microglia isolated from brain cortices of the neonatal male BALB/c mice, and M2 phenotype shifted microglia by IL-4 treatment were used for transplantation into the injured area after 24 h of ischemia induction. The behavioral outcomes evaluated by social interaction and Barnes tests and the levels of growth associated protein (GAP)-43 and inflammatory cytokine interleukin (IL)-1 protein were assessed by western blotting 7 days after cell transplantation. Animals in both of the microglia + pericytes and microglia M2 + pericytes transplanted groups showed better performance in social memory as well as enhanced spatial learning and memory compared to ischemic controls. Also, improved escape latency was only observed in microglia M2 + pericytes (p < 0.01) group compared to ischemic controls. GAP-43 showed significant protein expression in microglia + pericytes and microglia M2 + pericytes groups compared to the control group. Conversely, IL-1 levels diminished in all of the pericytes microglia + pericytes, and microglia M2 + pericytes groups compared to the ischemic controls. Current study highlights efficiency of M2 microglia and pericytes combinatory transplantation therapeutic role on relieving ischemic stroke outcomes.

Circulating H-FABP as a biomarker of frailty in patients with chronic heart failure.

Increased vulnerability to physiologic stressors, termed frailty, is a common occurrence in patients with chronic heart failure (CHF). However, the definite biomarkers to assess frailty in CHF patients are not known. Here, we assessed the frailty phenotype and its potential association with heart failure (HF) markers in CHF patients. We categorized controls (n = 59) and CHF patients (n = 80), the participants, into robust, pre-frail, and frail based on the cardiovascular health study (CHS) frailty index. The plasma levels of HF markers, including tumorigenicity 2 (s-ST2), galectin-3, and heart-type fatty acid binding protein (H-FABP), were measured and correlated with frailty phenotype and cardiac function. The levels of plasma s-ST2, galectin-3, and H-FABP were profoundly elevated in CHF patients. Conversely, the frailty index scores were significantly lower in ischemic and non-ischemic CHF patients versus controls. Of the assessed HF markers, only H-FABP was positively correlated (r2 = 0.07, P = 0.02) with the frailty score in CHF patients. Collectively, these observations suggest that circulating H-FABP may serve as a biomarker of frailty in CHF patients.

Influence of Age, Gender and Education Level on Executive Functions and Functioning in People with Stroke.

Alterations in mental functions are among the most frequent manifestations of stroke that have a direct impact on the patient's functionality. The objective of this study was to analyze the relationship of sociodemographic variables with the executive functions (EFs) of participants with right middle cerebral artery (MCA) stroke. A cross-sectional observational case-control study was conducted at the State Center for Brain Damage in Madrid, Spain. Fifty-eight subjects were recruited and divided into two groups. Each participant was administered the following: the FIM+FAM Functional Assessment Measure, the Lawton and Brody scale, The Trail-Making Test, the Zoo Map Test and the Hanoi Tower. Statistically significant differences (p < 0.05) were identified between participants with ischemic stroke and control in functional and EF functions, as well as between participants with hemorrhagic stroke and control. No statistically significant differences were found in the experimental group between subjects who had sustained ischemic and hemorrhagic stroke. No significant associations were identified between the variables age, gender and education level in relation to functionality and executive functions (p > 0.05) in people with stroke. People who have suffered a right cerebral artery stroke have deficiencies in the EFS, resulting in poorer performance of the activity of daily living, compared to healthy subjects of the same age, gender and education level. In the correlational analysis of the stroke participants, no significant associations were identified between the variables gender, age and education level in relation to functionality and EF.

Prevalence of Serum Vitamin-B12 Level and its Association Among the Patients of Transient Ischemic Attack/Ischemic Stroke

Objective: To determine the hospital-based prevalence of Vitamin-B12 deficiency in ischemic stroke patients and to compare the association of VitB-12 among patients of Ischemic stroke and controls. Study Design: Comparative study Place and Duration: Neurology and Medicine wards at the Liaquat University Hospital Hyderabad/Jamshoro. From Dec 2021 to June 2022 Methods: It was a non-probability convenience sample that comprised 263 individuals, of whom 132 were cases and 131 were controls. Among the participants, there were no missing data. The completion of a standardized questionnaire allowed for the collection of information about socio-demographic factors. Serum levels of VitB-12 were measured for all of the individuals aged 18 to 60 years old and of both genders Results: The gender distribution of cases and control groups, as well as the sociodemographic characteristics of the study population, have significant differences, in gender (p = 0.012) and socioeconomic status (p = 0.016). However, there was not a significant difference found in residence (0.82), nor age (0.46), in either of the groups. The average body mass index (BMI) was calculated to be 24.1 ± 6.1, while the average weight and height were determined to be 62.8 ± 12.3 kg and 158.62 ± 5.21cm, respectively. The incidence of ischemic stroke was recorded in 84 patients, whereas transient ischemic attack was documented in 48 patients. The percentage of instances that were affected by risk factors such as smoking, using paan gutka, or having high blood pressure was much greater than the percentage of cases that were affected by obesity, diabetic patients, or vegetarians. In all, there were 132 people out of 263 who were checked for their blood VitB-12 levels who were determined to have suffered from a stroke or transient ischemic attack (TIA). The cases comprised of both ischemic stroke or TIA. In a comparison of cases (ischemic stroke/TIA) and controls, it was shown that the patients had significantly lower mean levels of blood VitB-12 than the controls did (132.7+ 2.41 vs. 272.1+ 5.17 pg/mL respectively). Conclusion: The patients reported to neurology ward of Liaquat University hospital Hyderabad and Jamshoro, we found an unacceptably high burden of stroke and transient ischemic attack. There is a significant association between VitB-12 and ischemic stroke as compared to healthy controls. Keywords: Vitamin B-12, Ischemic Stroke, Risk Factors

Single-cell RNA sequencing reveals the transcriptomic landscape of kidneys in patients with ischemic acute kidney injury.

Ischemic acute kidney injury (AKI) is a common syndrome associated with considerable mortality and healthcare costs. Up to now, the underlying pathogenesis of ischemic AKI remains incompletely understood, and specific strategies for early diagnosis and treatment of ischemic AKI are still lacking. Here, this study aimed to define the transcriptomic landscape of AKI patients through single-cell RNA sequencing (scRNA-seq) analysis in kidneys. In this study, scRNA-seq technology was applied to kidneys from two ischemic AKI patients, and three human public scRNA-seq datasets were collected as controls. Differentially expressed genes (DEGs) and cell clusters of kidneys were determined. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, as well as the ligand-receptor interaction between cells, were performed. We also validated several DEGs expression in kidneys from human ischemic AKI and ischemia/reperfusion (I/R) injury induced AKI mice through immunohistochemistry staining. 15 distinct cell clusters were determined in kidney from subjects of ischemic AKI and control. The injured proximal tubules (PT) displayed a proapoptotic and proinflammatory phenotype. PT cells of ischemic AKI had up-regulation of novel pro-apoptotic genes including USP47 , RASSF4 , EBAG9 , IER3 , SASH1 , SEPTIN7 , and NUB1 , which have not been reported in ischemic AKI previously. Several hub genes were validated in kidneys from human AKI and renal I/R injury mice, respectively. Furthermore, PT highly expressed DEGs enriched in endoplasmic reticulum stress, autophagy, and retinoic acid-inducible gene I (RIG-I) signaling. DEGs overexpressed in other tubular cells were primarily enriched in nucleotide-binding and oligomerization domain (NOD)-like receptor signaling, estrogen signaling, interleukin (IL)-12 signaling, and IL-17 signaling. Overexpressed genes in kidney-resident immune cells including macrophages, natural killer T (NKT) cells, monocytes, and dendritic cells were associated with leukocyte activation, chemotaxis, cell adhesion, and complement activation. In addition, the ligand-receptor interactions analysis revealed prominent communications between macrophages and monocytes with other cells in the process of ischemic AKI. Together, this study reveals distinct cell-specific transcriptomic atlas of kidney in ischemic AKI patients, altered signaling pathways, and potential cell-cell crosstalk in the development of AKI. These data reveal new insights into the pathogenesis and potential therapeutic strategies in ischemic AKI.

M6A RNA methylation modification is involved in the disease course of heart failure

ABSTRACT We explored N 6-methyladenosine (m6A) RNA methylation as one of the gene regulatory mechanisms in heart failure (HF) biology. Understanding the different physiological mechanisms will facilitate the prevention and individualized treatment of HF. The Gene Expression Omnibus (GEO) database served as the source of the data. In GSE116250, differential analysis between ischemic cardiomyopathy (ICM), dilated cardiomyopathy (DCM) and controls yielded differentially expressed m6A regulators. Differential analysis between HF and controls in GSE131296 identifies m6A-modified genes and then performs enrichment analysis. Protein–protein interaction (PPI) network analysis was performed for the differentially expressed ICM- or DCM-associated genes in GSE116250 and GSE55296, respectively. Finally, the diagnostic genes for ICM and DCM were predicted using receiver operating characteristic (ROC) curve. YTHDC1, HNRNPC and HNRNPA2B1 were significantly downregulated in GSE116250 in DCM and ICM compared with controls. A total of 195 genes were identified in GSE131296 as subject to m6A alteration. These genes may play a role in HF through the MAPK signaling pathway and p53 signaling pathway. PPI network analysis identified CCL5, CXCR4 and CCL2 as key genes for ICM and IL-6 as a key gene for DCM. Through ROC curves, we identified m6A-modified APLP1, KLF2 as potential diagnostic genes for ICM, and m6A-modified FGF7, FREM1 and C14orf132 as potential diagnostic genes for DCM. Our findings support m6A modifying mechanisms in HF etiology that contribute to the treatment of HF. Thus, our data suggest that m6A methylation may be an interesting target for therapeutic intervention.

Direct and indirect activation of the adenosine triphosphate-sensitive potassium channel to induce spinal cord ischemic metabolic tolerance.

The mitochondrial adenosine triphosphate-sensitive potassium channel is central to pharmacologically induced tolerance to spinal cord injury. We hypothesized that both direct and nitric oxide-dependent indirect activation of the adenosine triphosphate-sensitive potassium channel contribute to the induction of ischemic metabolic tolerance. Spinal cord injury was induced in adult male C57BL/6 mice through 7minutes of thoracic aortic crossclamping. Pretreatment consisted of intraperitoneal injection 3 consecutive days before injury. Experimental groups were sham (no pretreatment or ischemia, n=10), spinal cord injury control (pretreatment with normal saline, n=27), Nicorandil 1.0mg/kg (direct and indirect adenosine triphosphate-sensitive potassium channel opener, n=20), Nicorandil 1mg/kg+carboxy-PTIO 1mg/kg (nitric oxide scavenger, n=21), carboxy-PTIO (n=12), diazoxide 5mg/kg (selective direct adenosine triphosphate-sensitive potassium channel opener, n=25), and DZ 5mg/kg+carboxy-PTIO 1mg/kg, carboxy-PTIO (n=23). Limb motor function was assessed using the Basso Mouse Score (0-9) at 12-hour intervals for 48hours after ischemia. Motor function was significantly preserved at all time points after ischemia in the Nicorandil pretreatment group compared with ischemic control. The addition of carboxy-PTIO partially attenuated Nicorandil's motor-preserving effect. Motor function in the Nicorandil+carboxy-PTIO group was significantly preserved compared with the spinal cord injury control group (P<.001), but worse than in the Nicorandil group (P=.078). Motor preservation in the diazoxide group was similar to the Nicorandil+carboxy-PTIO group. There was no significant difference between the diazoxide and diazoxide+carboxy-PTIO groups. Both direct and nitric oxide-dependent indirect activation of the mitochondrial adenosine triphosphate-sensitive potassium channel play an important role in pharmacologically induced motor function preservation.