Carbonic anhydrase inhibition for the management of cerebral ischemia: in vivo evaluation of sulfonamide and coumarin inhibitors.

Ischemia of brain areas is a global health problem, causing death or long-term disability. Current pharmacological options have limited impact on ischemic damages. Recently, a relationship between hypoxia and carbonic anhydrase (CA) over-expression has been highlighted suggesting CA inhibition as a possible target. This study aimed to evaluate the pharmacological profile of sulfonamide and coumarin CA inhibitors in rats underwent permanent middle cerebral artery occlusion (pMCAO). The neurological score of pMCAO rats was dramatically reduced 24 h after occlusion. Repeated subcutaneous injections of the CA inhibitors 4 and 7 (1 mg kg(-1)) were able to increase the neurological score by 40%. Compound 7 showed the tendency to reduce the volume of hemisphere infarction. The standard CA inhibitor acetazolamide was ineffective. The properties of novel CA inhibitors to improve neurological functionalities after cerebral ischemic insult are shown. The CA involvement in cerebral hypoxic phenomena deserves deeper investigations.

Neurological scoring and gait kinematics to assess functional outcome in an ovine model of ischaemic stroke.

Background: Assessment of functional impairment following ischaemic stroke is essential to determine outcome and efficacy of intervention in both clinical patients and pre-clinical models. Although paradigms are well described for rodents, comparable methods for large animals, such as sheep, remain limited. This study aimed to develop methods to assess function in an ovine model of ischaemic stroke using composite neurological scoring and gait kinematics from motion capture. Methods: Merino sheep (n = 26) were anaesthetised and subjected to 2 hours middle cerebral artery occlusion. Animals underwent functional assessment at baseline (8-, 5-, and 1-day pre-stroke), and 3 days post-stroke. Neurological scoring was carried out to determine changes in neurological status. Ten infrared cameras measured the trajectories of 42 retro-reflective markers for calculation of gait kinematics. Magnetic resonance imaging (MRI) was performed at 3 days post-stroke to determine infarct volume. Intraclass Correlation Coefficients (ICC's) were used to assess the repeatability of neurological scoring and gait kinematics across baseline trials. The average of all baselines was used to compare changes in neurological scoring and kinematics at 3 days post-stroke. A principal component analysis (PCA) was performed to determine the relationship between neurological score, gait kinematics, and infarct volume post-stroke. Results: Neurological scoring was moderately repeatable across baseline trials (ICC > 0.50) and detected marked impairment post-stroke (p < 0.05). Baseline gait measures showed moderate to good repeatability for the majority of assessed variables (ICC > 0.50). Following stroke, kinematic measures indicative of stroke deficit were detected including an increase in stance and stride duration (p < 0.05). MRI demonstrated infarction involving the cortex and/or thalamus (median 2.7 cm3, IQR 1.4 to 11.9). PCA produced two components, although association between variables was inconclusive. Conclusion: This study developed repeatable methods to assess function in sheep using composite scoring and gait kinematics, allowing for the evaluation of deficit 3 days post-stroke. Despite utility of each method independently, there was poor association observed between gait kinematics, composite scoring, and infarct volume on PCA. This suggests that each of these measures has discreet utility for the assessment of stroke deficit, and that multimodal approaches are necessary to comprehensively characterise functional impairment.

Introducing Adam's Scale of Posterior Stroke (ASPOS): A Novel Validated Tool to Assess and Predict Posterior Circulation Strokes.

BACKGROUND: Assessing the severity of posterior circulation strokes, due to the variety of symptoms, is a significant clinical problem. Current clinimetric scales show lower accuracy in the measurement of posterior stroke severity, compared with that of anterior strokes. The aim of the study was to design a validated tool, termed Adam's Scale of Posterior Stroke (ASPOS), for better assessment and prediction of posterior stroke. METHODS: This prospective, observational study involved 126 posterior circulation ischemic stroke subjects. Four researchers, previously trained in ASPOS, randomized the stroke severity using a novel tool and other appropriate stroke scales (The National Institute of Health Stroke Scale-NIHSS, modified Rankin Scale-mRS, Glasgow Coma Scale, Barthel Index, or Israeli Vertebrobasilar Stroke Scale-IVBSS) to assess the psychometric properties, reliability, and validity of ASPOS and investigate its predictive value. RESULTS: ASPOS reached a Cronbach's alpha coefficient of 0.7449, indicating good internal consistency. The Bland-Altman analysis showed a good coefficient of repeatability (CR) of 0.46, a 95% confidence interval (CI) of 0.41-0.53, and excellent intraclass correlation coefficients or weighted kappa values (>0.90), reflecting high reliability and reproducibility. Highly significant correlations with other scales confirmed the construct and predictive validity of ASPOS. A total ASPOS score of three points indicated a significantly increased probability of severe stroke based on the NIHSS, compared to a total ASPOS of 1-2 points (odds ratio (OR) 141; 95% CI: 6.72-2977.66; p = 0.0014). CONCLUSIONS: We developed a novel, valid, and reliable tool to assess posterior circulation strokes. This can contribute to a more comprehensive estimation of posterior stroke and, additionally, due to its predictive properties, it can be used to more accurately select candidates for specific treatments.

Neurological Exam in Rats Following Stroke and Traumatic Brain Injury.

Using the appropriate model for testing neurological symptoms in rats is essential for the assessment of functional outcome. A number of tests have been developed to quantify the severity of neurological deficits. These tests should meet criteria such as validity, specificity, sensitivity, and utility. Although analysis of motor function shows homology in primates and rodents, the total neurological exam scores may not always reflect the clinical outcome. Therefore, the selection of the appropriate tests has critical importance when evaluating therapeutic strategies. This chapter describes Toklu's modified neurological exam score method which can be used practically to assess neurological symptoms following traumatic brain injury (TBI) and stroke. The method is a combination of balance, muscle strength, coordination, and reflex.

Balinese Cultivar of Purple Sweet Potato Improved Neurological Score and BDNF and Reduced Caspase-Independent Apoptosis among Wistar Rats with Ischemic Stroke.

BACKGROUND: Ischemic stroke occurs due to the abrupt occlusion in the brain which leads to neuronal death. Neuronal death in ischemic stroke is due to increase production of reactive oxygen species (ROS). Neuronal death occurs via necrosis and apoptosis mechanisms. Apoptosis can either occur via extrinsic or intrinsic pathway. Meanwhile, the intrinsic pathway can be caspase-dependent or independent. Anthocyanin is a natural pigment with antioxidant, anti-inflammatory, anti-cancer, and neuroprotective properties. Balinese cultivate of purple potato extract contains a high level of anthocyanin and has been proven for its antioxidant activity. AIM: Antioxidant effect of Balinese cultivates purple potato extract has not been studied on an animal model with ischemic stroke. Accordingly, we would like to study the effect of antioxidant properties from Balinese cultivate of purple potato extract by assessing the neurological score, BNDF concentration, and caspase-independent apoptosis by measuring AIF concentration on Wistar rats with ischemic stroke. METHODS: This was an experimental study using male Wistar rats age between 12-14 weeks weigh between 200 to 250 g. RESULTS: This study demonstrated a significant difference of neurological score on day 3 among control versus treatment groups. Balinese cultivate of purple potato extract markedly reduced AIF, increased BDNF, and suppressed apoptosis among treatment group when compared with the control group. CONCLUSION: We have proven the efficacy of antioxidant activity of anthocyanin derived from Balinese cultivar of purple sweet potato by elevated AIF levels, lower apoptosis rate, improved neurological score on day-3 to day-7 post-stroke, as well as increased BDNF levels.

Genetic deletion of mGlu2 metabotropic glutamate receptors improves the short-term outcome of cerebral transient focal ischemia.

We have recently shown that pharmacological blockade of mGlu2 metabotropic glutamate receptors protects vulnerable neurons in the 4-vessel occlusion model of transient global ischemia, whereas receptor activation amplifies neuronal death. This raised the possibility that endogenous activation of mGlu2 receptors contributes to the pathophysiology of ischemic neuronal damage. Here, we examined this possibility using two models of transient focal ischemia: (i) the monofilament model of middle cerebral artery occlusion (MCAO) in mice, and (ii) the model based on intracerebral infusion of endothelin-1 (Et-1) in rats. Following transient MCAO, mGlu2 receptor knockout mice showed a significant reduction in infarct volume and an improved short-term behavioural outcome, as assessed by a neurological disability scale and the "grip test". Following Et-1 infusion, Grm2 gene mutated Hannover Wistar rats lacking mGlu2 receptors did not show changes in the overall infarct volume as compared to their wild-type counterparts, although they showed a reduced infarct area in the agranular insular cortex. Interestingly, however, mGlu2 receptor-deficient rats performed better than wild-type rats in the adhesive tape test, in which these rats did not show the laterality preference typically observed after focal ischemia. These findings support the hypothesis that activation of mGlu2 receptors is detrimental in the post-ischemic phase, and support the use of mGlu2 receptor antagonists in the experimental treatment of brain ischemia.

Neuroprotective effect of epidermal growth factor plus growth hormone-releasing peptide-6 resembles hypothermia in experimental stroke.

BACKGROUND: Combined therapy with epidermal growth factor (EGF) and growth hormone-releasing peptide 6 (GHRP-6) in stroke models has accumulated evidence of neuroprotective effects from several studies, but needs further support before clinical translation. Comparing EGF + GHRP-6 to hypothermia, a gold neuroprotection standard, may contribute to this purpose. OBJECTIVES: The aims of this study were to compare the neuroprotective effects of a combined therapy based on EGF + GHRP-6 with hypothermia in animal models of (a) global ischemia representing myocardial infarction and (b) focal brain ischemia representing ischemic stroke. METHODS: (a) Global ischemia was induced in Mongolian gerbils by a 15-min occlusion of both carotid arteries, followed by reperfusion. (b) Focal brain ischemia was achieved by intracerebral injection of endothelin 1 in Wistar rats. In each experiment, three ischemic treatment groups - vehicle, EGF + GHRP-6, and hypothermia - were compared to each other and to a sham-operated control group. End points were survival, neurological scores, and infarct volume. RESULTS: (a) In global ischemia, neurological score at 48-72 h, infarct volume, and neuronal density of hippocampal CA1 zone in gerbils treated with EGF + GHRP-6 were similar to the hypothermia-treated group. (b) In focal ischemia, the neurologic score and infarct volume of rats receiving EGF + GHRP-6 were also similar to animals in the hypothermia group. DISCUSSION: With hypothermia being a good standard neuroprotectant reference, these results provide additional proof of principle for EGF and GHRP-6 co-administration as a potentially neuroprotective stroke therapy.

Clinical hypoxic-ischemic encephalopathy score of the Iberoamerican Society of Neonatology (Siben): A new proposal for diagnosis and management / Escore clínico de encefalopatia hipóxico-isquêmica da Sociedad Iberoamericana de Neonatología (Siben): uma nova proposta para seu diagnóstico e manejo perinatal

Summary Hypoxic ischemic encephalopathy is a major complication of perinatal asphyxia, with high morbidity, mortality and neurologic sequelae as cerebral palsy, mostly in poor or developing countries. The difficulty in the diagnosis and management of newborns in these countries is astonishing, thus resulting in unreliable data on this pathology and bad outcomes regarding mortality and incidence of neurologic sequelae. The objective of this article is to present a new clinical diagnostic score to be started in the delivery room and to guide the therapeutic approach, in order to improve these results.

Resumo A encefalopatia hipóxico-isquêmica é a principal complicação da asfixia perinatal, com alta morbidade, mortalidade e incidência de sequelas neurológicas, como a paralisia cerebral, principalmente em países pobres e/ou em desenvolvimento. Nessas regiões, as dificuldades no diagnóstico e no manejo desses recém-nascidos é surpreendente, o que resulta em dados pouco confiáveis e em péssimos desfechos tanto no que se refere à mortalidade como à incidência de sequelas neurológicas. O objetivo deste artigo é apresentar um novo escore para o diagnóstico clínico ser iniciado na sala de parto e uma abordagem terapêutica com o intuito de melhorar esses resultados.

Research progress of prevention and cue effects of caffeoylquinic acids from Erigeron breviscapus on ischemic stroke / 中草药

This paper summarized the chemical structures and amounts of caffeoylquinic acids in Erigeron breviscapus, as well as its prevention and cure effects on ischemic stroke in clinical and possible pharmacological mechanism. The results showed 22 caffeoylquinic acids reported from E. breviscapus, accounting for 29% of all compounds from this herb; The average content of total polyphenols was 36.93%, and more than 95% of components in Erigeron Breviscapus Injection are caffeoylquinic acids, higher than that of scutellarin. Several high quality clinical studies confirmed that Erigeron Breviscapus Injection enhanced treatment performance and improve the neurological score in the treatment after ischemic stroke and had good safety. In pharmacological research, caffeoylquinic acid compounds display anti-oxidant, anti-free radical, anticoagulation, and anti-fibrosis effects, which can protect neuro, vascular endothelial cells, glial cells, and astrocytes. They are also able to inhibit inflammatory, suppress cytokines IL-1, TNF-α, and enhance SOD & GSH-Px, which play a role in different treatment stages of ischemic stroke. So, caffeoylquinic acid is a kind of important chemical in E. breviscapus.

Establish and optimize the model of experimental allergic encephalomyelitis in mice / 中华行为医学与脑科学杂志

Objective To explore the appropriate dosage of drugs inducing experimental allergic en cephalomyelitis (EAE) in mice,and evaluate the modified model mice.Methods Different doses of myelin oligodendrocyte glycoprotein (MOG35-55:200 μg,100 μg,50 μg,25 μg),together with different doses of inactivation of mycobacterium tuberculosis (H37RA:800 μg,250 μg,100 μg) and pertussis toxin (500 ng,200 ng),were used to induce the EAE model.After immunized,the clinical disease severity of EAE mice was measured by the standard EAE grading clinical score daily.The open field test was used to detect the locomotion of mice.The Western blot and immunofluorescence were used to detect the level of myelin basic proteins (MBP) in different brain regions of mice.Results Compared with the EAE mice induced with high-dose drugs,the mice with low-dose drugs (25 μg MOG35-55,100 μg H37RA,200 ng pertussis toxin) had low neu rological scores.And they displayed normal locomotion compared with the control mice (day 16:group EAE (8.885±0.772) cm/s vs control group (8.933±0.567) cm/s,P＞0.05;day 31:group EAE (11.130±0.630) cm/s vs control group (10.670±0.959) cm/s,P＞0.05;day 55:group EAE (7.686±0.428) cm/s vs control group (8.313±0.918) cm/s,P＞0.05).Moreover,there was a significant decrease of MBP in the parahippocampal cortex (PHC) and fimbria-fornix of EAE mice induced with low-dose of drugs (PHC:group EAE (0.369±0.096) vs control group (1.000±0.163),P＜0.05;fimbria-fomix:group EAE (0.494±0.071) vs control group (1.000±0.143),P＜0.05).Conclusion The EAE mice induced with low-dose drugs(25 μg MOG35-55,100 μg H37RA,200 ng pertussis toxin) have low neurological scores,normal locomotion,and myelin impairment in the central neuronal system.And it can be used in the cognitive behavioral research of demyelination disease,such as multiple sclerosis.

CRMP2 alleviates neurological deficit by reducing neuron apoptosis in rats after cerebral ischemia/reperfusion injury / 中国药理学通报

Aim To investigate the influence of the overexpression of CRMP2 on neural cell apoptosis after ischemia reperfusion injury in rats and its possible mechanism. Methods A total of 192 male adult SD rats were divided into four groups: sham group, cere-bral ischemia/reperfusion group( MCAO group) , cere-bral ischemia with blank plasmid control group( MCAO+GFP group ) , cerebral ischemia with CRMP2 eu-karyotic plasmid group ( MCAO + CRMP2/GFP group) . One day after injecting eukaryotic plasmid, the rats were operated for 120-min ischemia through MCA occlusion and reperfused. At 48 h and 1 wk, the expression of CRMP2 , p53 , Caspase-3 , Caspase-8 and BCL2 in brain tissue was tested by RT-PCR and West-ern blot. Apoptotic cells were observed by TUNEL test. TTC staining was use to detect cerebral infarction volume. The neural function of the rats were also eval-uated. Results Compared with the sham group, the expression levels of CRMP2 and BCL2 in MCAO group and MCAO +GFP group were significantly decreased ( P <0. 01 ) , while p53 , Caspase-3 , Caspase-8 and TUNEL positive cells were elevated(P<0. 01). Inter-vention of CRMP2 eukaryotic plasmid resulted in the increased expression of CRMP2 and BCL2 ( P<0. 01 ) and the decreased p53 , Caspase-3 and Caspase-8 ex-pression. In TUNEL test, overexpression of CRMP2 obviously decreased the number of TUNEL positive cells(P<0. 01). The expression of BDNF was upregu-lated after cerebral ischemic injury ( P<0. 01 ) , while overexpression of CRMP2 increased BDNF more signif-icantly ( P <0. 01 ) . TTC staining showed cerebral in-farction Volume of MCAO + CRMP2/GFP group was obviously decreased ( P <0. 01 ) , and neurologic defi-cits were significantly improved ( P <0. 01 ) . Conclu-sion The overexpression of CRMP2 reduces nerve cell apoptosis possibly by regulating the mitochondrial ap-optosis pathway after cerebral ischemia/reperfusion in-jury to protect nervous system.

Effects of Instant Hyperbaric Oxygen Therapy on Brain Edema and Expression of Aquaporin-4 after Cerebral Ischemia in Rats / 中国康复理论与实践

Objective To observe the effects of instant hyperbaric oxygen (HBO) therapy on brain edema and aquaporin-4 (AQP4) ex-pression after cerebral ischemia in rats. Methods 18 healthy adult male Sprague-Dawley rats were randomly divided into sham group (n=6), control group (n=6) and HBO group (n=6). Middle cerebral artery occlusion model was established with modified Longa method in the con-trol group and HBO group. HBO was administered after cerebral ischemia immediately in HBO group for 60 min. They were observed with the neurological function score, brain tissue water content and AQP4 experssion 6 h after operation. Results Compared with the control group, the neurological score in HBO group improved significantly (P<0.05), while the brain tissue water content and brain AQP4 expres-sion decreased (P<0.05). Conclusion HBO may improve the neurological function and brain edema after cerebral ischemia, which may re-late with inhibiting the expression of AQP4.

Effects of Instant Hyperbaric Oxygen Therapy on Brain Edema and Expression of Aquaporin-4 after Cerebral Ischemia in Rats / 中国康复理论与实践

@#Objective To observe the effects of instant hyperbaric oxygen (HBO) therapy on brain edema and aquaporin-4 (AQP4) expression after cerebral ischemia in rats. Methods 18 healthy adult male Sprague-Dawley rats were randomly divided into sham group (n=6),control group (n=6) and HBO group (n=6). Middle cerebral artery occlusion model was established with modified Longa method in the control group and HBO group. HBO was administered after cerebral ischemia immediately in HBO group for 60 min. They were observed with the neurological function score, brain tissue water content and AQP4 experssion 6 h after operation. Results Compared with the control group, the neurological score in HBO group improved significantly (P<0.05), while the brain tissue water content and brain AQP4 expression decreased (P<0.05). Conclusion HBO may improve the neurological function and brain edema after cerebral ischemia, which may relate with inhibiting the expression of AQP4.