Investigating the mechanism of interactive regulation of B-cell lymphoma-2/Beclin 1 through electroacupuncture intervention during reperfusion in myocardial ischemia-reperfusion injury in a rat model.

To observe the regulation of B-cell lymphoma-2 (Bcl-2)/Beclin 1 interaction through electroacupuncture (EA) intervention during reperfusion and to investigate the EA mechanism of apoptosis-autophagy interactive regulation against myocardial ischemia-reperfusion injury (MIRI). A total of 48 adult Sprague Dawley (SD) rats were randomly divided into the sham-operated group (group Sham), the model group (group Model), the EA group (group EA), and the JNK inhibitor (SP600125) group (group JNK), with 12 rats in each group. Biospecimens were collected randomly from six rats in each group four hours after reperfusion. Evans Blue and triphenyl tetrazolium chloride double-staining were applied to observe each group's myocardial damage area and risk area. We collected 4 ml of blood by abdominal aortic method to detect serum troponin cTnI level by enzyme-linked immunosorbent assay (ELISA). For the remaining six in each group, a part of myocardial tissue below the ligation line was stored in 4% paraformaldehyde for immunohistochemistry and TUNEL staining; the other amount of myocardial tissue was detected by Western blotting to determine the expression levels of Bcl-2, Beclin1, and the phosphorylation levels of Thr69, Ser70, and Ser87 in Bcl-2. In results: electroacupuncture (EA) intervention during reperfusion significantly reduced the myocardial infarction area, cTnI level, and myocardial apoptosis, upregulated Bcl-2 expression, downregulated Beclin 1 expression and inhibited phosphorylation levels of Thr69, Ser70, and Ser87 in Bcl-2. We concluded that EA effectively inhibited apoptosis by upregulating Bcl-2 expression and inhibiting the phosphorylation of Thr69, Ser70, and Ser87 in Bcl-2. This reduced the separation of Bcl-2 and Beclin 1, restrains excessive autophagy, alleviates MIRI, and has a protective effect on myocardial tissue.

Hawthorn revisited: time- and dose-dependent cardioprotective action of WS-1442 special extract in the reperfusion-induced arrhythmia model in rats in vivo.

Standardized WS-1442 extract from Crataegus oxycantha (hawthorn) leaves and berries is one of the most widely studied preparations received from hawthorn. This popular substance is known from its positive influence on the cardiovascular system. The current research aimed to evaluate the optimal dose of standardized WS-1442 extract and the most beneficial period for its use. The study analysis was based on experiments previously conducted on male Sprague-Dawley rats (n = 152). The animals were divided into subgroups to examine the relationship between the dose-dependent (n = 96) and time-dependent (n = 56) effects of the mentioned extract. The research was performed based on the modified early reperfusion-induced arrhythmias model in vivo. The following parameters were assessed during the study: efficiency of mortality index reduction, reduction of ventricular arrhythmias incidences as well as the influence of standardized WS-1442 extract on hemodynamic parameters and amount of biochemical marker of cardiac tissue damage (creatine kinase). The current study revealed the dose- and time-dependent cardioprotective effect of standardized WS-1442 extract. It was expressed by mortality index reduction, decrease in the incidence and duration of severe ventricular arrhythmias and decline in the total amount of creatine kinase. Analyzed data coming from a model of reperfusion-induced arrhythmias in rats suggests that standardized WS-1442 extract is a potent cardioprotective agent whose action depends on both dose and intake time.

Effects of Shaoyao-Gancao Decoction on Infarcted Cerebral Cortical Neurons: Suppression of the Inflammatory Response following Cerebral Ischemia-Reperfusion in a Rat Model.

The mechanisms by which Shaoyao-Gancao decoction (SGD) inhibits the production of inflammatory cytokines in serum and brain tissue after cerebral ischemia-reperfusion (CI-RP) in rats were investigated. A right middle cerebral artery occlusion was used to induce CI-RP after which the rats were divided into model (n = 39), SGD (n = 28), clopidogrel (n = 25) and sham operated (n = 34) groups. The Bederson scale was used to evaluate changes in behavioral indices. The levels of IL-1ß, TNF-α, MCP-1, IL-10, RANTES, VEGF, and TGF-ß1 in the serum and infarcted brain tissues were measured. Nissl body and immunohistochemical staining methods were used to detect biochemical changes in neurons, microglial cells, and astrocytes. Serum levels of VEGF, TNF-α, MCP-1, IL-1ß, and IL-10 increased significantly 24 h after CI-RP. In brain tissue, levels of TNF-α and IL-1ß significantly increased 24 h after CI-RP, whereas levels of TGF-ß1 and MCP-1 were significantly higher 96 h after CI-RP (P < 0.05). SGD or clopidogrel after CI-RP reduced TNF-α and IL-1ß levels in brain tissue and serum levels of MCP-1, IL-1ß, and IL-10. SGD increased the number of NeuN-positive cells in infarcted brain tissue and reduced the number of IBA1-positive and GFAP-positive cells. The efficacy of SGD was significantly higher than that of clopidogrel.

Electroacupuncture modulates stromal cell-derived factor-1α expression and mobilization of bone marrow endothelial progenitor cells in focal cerebral ischemia/reperfusion model rats.

Stromal cell-derived factor-1α(SDF-1α) plays a crucial role in regulating the mobilization, migration and homing of endothelial progenitor cells(EPCs). Electroacupuncture(EA), a modern version of Traditional Chinese Medicine, can improve neurological recovery and angiogenesis in cerebral ischemic area. This study aimed to investigate the effects of electroacupuncture(EA) on the mobilization and migration of bone marrow EPCs and neurological functional recovery in rats model after focal cerebral ischemia/reperfusion and the potentially involved mechanisms. Sprague-Dawley rats received filament occlusion of the right middle cerebral artery for 2h followed by reperfusion for 12h, 1d, 2d, 3d, 7d respectively. Rats were randomly divided into sham group, model group and EA group. After 2h of the reperfusion, EA was given at the "Baihui" (GV 20)/Siguan ("Hegu" (LI 4)/"Taichong" (LR 3)) acupoints in the EA group. Modified neurological severity score (mNSS) was used to assess the neurological functional recovery. EPCs number and SDF-1α level in bone marrow(BM) and peripheral blood(PB) were detected by using fluorescence-activated cell sorting (FACS) analysis and quantitative real time polymerase chain reaction (qRT-PCR) respectively. An mNSS test showed that EA treatment significantly improved the neurological functional outcome. EPCs number in PB and BM were obviously increased in the EA group. After cerebral ischemia, the SDF-1α level was decreased in BM while it was increased in PB, which implied a gradient of SDF-1α among BM and PB after ischemia. It suggested that the forming of SDF-1α concentration gradient can induce the mobilization and homing of EPCs. Eletroacupuncture as a treatment can accelerate and increase the forming of SDF-1α concentration gradient to further induce the mobilization of EPCs and angiogenesis in ischemic brain and improve the neurological function recovery.

The Effects of Vaccinium myrtillus Extract on Hamster Pial Microcirculation during Hypoperfusion-Reperfusion Injury.

INTRODUCTION: The present study was aimed to assess the in vivo hamster pial microvessel alterations due to 30 min transient bilateral common carotid artery occlusion (BCCAO) and reperfusion (60 min); moreover, the neuroprotective effects of Vaccinium myrtillus extract, containing 34.7% of anthocyanins, were investigated. MATERIALS AND METHODS: Two groups of male hamsters were used: the first fed with control diet and the other with Vaccinium myrtillus supplemented diet. Hamster pial microcirculation was visualized by fluorescence microscopy through an open cranial window. Pial arterioles were classified according to Strahler's method. RESULTS: In age-matched control diet-fed hamsters, BCCAO caused a decrease in diameter of all arterioles. At the end of reperfusion, the reduction of diameter in order 3 arterioles was by 8.4 ± 3.1%, 10.8 ± 2.3% and 12.1 ± 1.1% of baseline in the 2, 4 and 6 month control diet-fed hamsters, respectively. Microvascular permeability and leukocyte adhesion were markedly enhanced, while perfused capillary length (PCL) decreased. The response to acetylcholine and papaverine topical application was impaired; 2'-7'-dichlorofluoresceine-diacetate assay demonstrated a significant ROS production. At the end of BCCAO, in age-matched Vaccinium myrtillussupplemented diet-fed hamsters, the arteriolar diameter did not significantly change compared to baseline. After 60 min reperfusion, order 3 arterioles dilated by 9.3 ± 2.4%, 10.6 ± 3.1% and 11.8 ± 2.7% of baseline in the 2, 4 and 6 month Vaccinium myrtillus supplemented diet-fed hamsters, respectively. Microvascular leakage and leukocyte adhesion were significantly reduced in all groups according to the time-dependent treatment, when compared with the age-matched control diet-fed hamsters. Similarly, the reduction in PCL was progressively prevented. Finally, the response to acetylcholine and papaverine topical application was preserved and there was no significant increase in ROS production in all groups. CONCLUSIONS: In conclusion, Vaccinium myrtillusextract protected pial microcirculation during hypoperfusion-reperfusion, preventing vasoconstriction, microvascular permeability, leukocyte adhesion, reduction in PCL and preserving the endothelium function.

Relative neuroprotective effects hyperbaric oxygen treatment and TLR4 knockout in a mouse model of temporary middle cerebral artery occlusion.

PURPOSE: To examine the effects of hyperbaric oxygen (HBO) therapy and knockout of toll-like receptor 4 (TLR4) on the outcome of temporary middle cerebral artery occlusion (MCAO) in a mouse model. MATERIALS AND METHODS: MCAO was induced in anesthetized male C57Bl/6 mice (WT) and TLR4 knockout mice (TLR4(-/-)) using an intra-arterial filament method. After 30 or 90 min, the filament was removed, and the mice were given either no treatment (WT and TLR4(-/-) groups) or HBO (WT only). Mice were euthanized 24 h after MCAO, and the brain infarct area was examined using 2,3,5-triphenyltetrazolium chloride (TTC) staining. RESULTS: In the WT group, without treatment, lesion volume was 120 ± 13 mm(3) in the mice subjected to 30 min' MCAO and 173 ± 23 mm(3) in the mice subjected to 90 min' MCAO. Respective values with HBO treatment were 66.5 ± 36.7 mm(3) and 53.2 ± 17.2 mm(3). The difference was significant only for 90-minute MCAO (p < 0.01, nonparametric test). In the TLR4(-/-) group (all untreated), lesion volume was 95.9 ± 17.9 after 90 min of MCAO, which was significantly lower than in the untreated WT animals (p < 0.05, nonparametric test). CONCLUSIONS: A single treatment of HBO immediately after MCAO followed by 24 h' reperfusion significantly reduces edema and may improve perfusion. TLR4 knockout protects mice from MCAO damage, but to a lesser extent than HBO treatment.

By pass arterial femorodistal bajo anestesia local / Distal femoral bypass under local anaesthesia

Introducción: El objetivo de este trabajo es transmitir nuestra experiencia en by pass arterial femorodistal bajo anestesia local para pacientes de alto riesgo. Material y métodos De enero a mayo de 2010 hasta 8 pacientes han sido intervenidos en nuestro centro de cirugía de revascularización en miembros inferiores bajo anestesia local. Las características comunes de los pacientes eran la edad avanzada, cardiopatía isquémica crónica en tratamiento con antiagregantes plaquetarios y la enfermedad pulmonar obstructiva crónica (EPOC).Resultados Los 8 pacientes (100%) toleraron de forma adecuada el procedimiento sin recurrir a sedación ni a procedimientos anestésicos invasivos. La arteria receptora fue la tibial posterior en 6 de ellos (75%) y la poplítea y peronea en uno (12%). En 7 casos logramos permeabilidad precoz del injerto y uno presentó trombosis precoz con amputación supracondílea; la técnica empleada fue femoropoplíteo en un caso, femorotibial posterior en 6 casos y poplíteo-peroneo en otro. El injerto utilizado fue la vena safena mayor, invertida en un paciente (12%) e in situ en 7 (88%) con valvulotomo BARD™, 3mm. Las complicaciones postoperatorias fueron del 0% para hemorragia, infección y muerte; en 3 pacientes (37%) amputación menor y, tras un periodo de seguimiento medio de 3 meses (1-4 meses), 7 casos están libres de amputación mayor. Conclusiones La revascularización del territorio distal de los miembros inferiores puede llevarse a cabo mediante anestesia local de forma segura y eficaz, sin necesidad de retirar antiagregantes ni asumir los riesgos de la anestesia general. La anatomía desfavorable (obesidad) puede limitar el procedimiento (AU)

Introduction: The aim of this article is to present our experience in performing distal femoralby pass under local anaesthesia for high risk patients. Material and methods: Lower limb revascularisation surgery under local anaesthesia was performed on 8 patients in our centre between January and May 2010. The common characteristics of the patients were, advanced age, chronic is chaemic heart disease on antiplatelet treatment, and chronic obstructive pulmonary disease (COPD). Results: All 8 patients (100%) tolerated the procedure well without having to resort to sedation or invasive an aesthetic procedures. The receiving artery was the posterior tibialin 6 cases (75%) and the popliteal and peroneal in 1 (12%). Early patency of the graft was achieved 7 patients and 1 had early thrombosis with a supracondylar amputation. The technique used was femoral-popliteal in 1 case, femoral-posterior tibial in 6 cases, and popliteal-peroneal in 1 case. The saphenous vein was the graft used, inverted in 1 patient(12%), and in situ in 7 (88%) with a 3 mm BARDTM valvotomy. There were no post-operative complications as regards haemorrhage, infections or death. A minor amputation was performed on 3 patients (37%), and after a mean of 3 months (1-4 months) follow-up,7 cases were free of major amputation. Conclusions: Revascularisation of the distal zone of the lower limbs can be safely and effectively performed using local anaesthesia, avoiding the risks of general anaesthesia and without the need to stop antiplatelet treatment. Anatomical problems (obesity) may limit the procedure (AU)

Neuroprotective effects of Schisandrin B against transient focal cerebral ischemia in Sprague-Dawley rats.

Fruits of Schisandra have been traditionally used in East Asia for the treatment of dyspnea, cough, dysentery, insomnia, tonic-clonic seizures, and amnesia. Schisandrin B, a dibenzocyclooctadiene derivative isolated from Fructus Schisandrae, has been shown to produce antioxidant effect on rodent liver and heart. In the present study, we investigated the neuroprotective effects of Schisandrin B, a constituent drug of the fruit of Schisandra, against focal cerebral ischemia in rats. Schisandrin B (10, 30 mg/kg, i.p.) was twice administered 30 min before the onset of ischemia and 2h after reperfusion. Schisandrin B 10 and 30 mg/kg treated groups showed infarct volumes reduced by 25.7% and 53.4%, respectively, 2h after occlusion. Also, Schisandrin B treated animal treatment abrogated protein expression of TNF-α and IL-1ß and degradation of MMP-2 and MMP-9 in ischemic hemispheres. These results suggest that Schisandrin B treatment provides a neuroprotective effect to rats after transient focal cerebral ischemia by inhibiting inflammation and by protecting against metalloproteinase degradation.

Evaluation of the neuroprotective effect of minocycline in a rabbit spinal cord ischemia model.

OBJECTIVE: To investigate whether postischemic administration of minocycline attenuates hind-limb motor dysfunction and gray and white matter injuries after spinal cord ischemia. DESIGN: A prospective, randomized, laboratory investigation. SETTING: Laboratory in university, single institution. PARTICIPANTS: Male New Zealand White rabbits. INTERVENTION: Spinal cord ischemia was induced by an occlusion of the infrarenal aorta for 15 minutes. The groups were administered minocycline 1 hour after reperfusion (M-1; n = 8), minocycline 3 hours after reperfusion (M-3; n = 8), saline 1 hour after reperfusion (control [C]; n = 8), or saline and no occlusion (sham; n = 4). Minocycline was administered intravenously at 10 mg/kg 6 times at 12-hour intervals until 60 hours after the initial administration. MEASUREMENT AND MAIN RESULTS: Hind-limb motor function was assessed using the Tarlov score. For histologic assessments, gray and white matter injuries were evaluated 72 hours after reperfusion using the number of normal neurons and the percentage of areas of vacuolation, respectively. Motor function 72 hours after reperfusion was significantly better in group M-1 than in group C. The number of neurons in the anterior horn was significantly larger in group M-1 than in groups M-3 or C but did not differ significantly between groups M-3 and C. No significant difference was noted in the percentage of areas of vacuolation among the ischemia groups. CONCLUSIONS: Minocycline administration beginning at 1 hour after reperfusion improved hind-limb motor dysfunction and attenuated gray matter injury in a rabbit spinal cord ischemia model.

The effects of hypothermia and L-arginine on skeletal muscle function in ischemia-reperfusion injury.

OBJECTIVES: Ischemia-reperfusion (I/R) injury can have detrimental effects on skeletal muscle. We have shown that vessel permeability can be minimized in a hypothermic setting and also by administering the nitric oxide synthase (NOS) stimulator, L-arginine, at physiologic temperatures. The purpose of this study was to examine and compare skeletal muscle contractility after an I/R insult during hypothermic conditions, warm conditions, and also with the administration of L-arginine at physiologic temperatures. We hypothesized that hypothermia and L-arginine administration will also demonstrate protective effects to skeletal muscle contractility. METHODS: Using Sprague-Dawley rats, the extensor digitorum longus muscle was rotated on its vascular pedicle to a thermo-controlled stage. Ischemia was established using an atraumatic femoral artery tourniquet. Reperfusion was performed under control and experimental conditions including local hypothermia and intravenous L-arginine. After harvesting experimental muscles, contractility was then quantified by using a tissue bath stimulator with force transducers. RESULTS: Warm reperfusion resulted in marked decrease in muscle contractility compared with sham animals. Local hypothermia showed statistically significant preservation of contractility compared with the sham group. This protective effect was recapitulated by the application of NOS inducers (L-arginine) at warm conditions. CONCLUSIONS: These findings demonstrate that hypothermia and L-arginine are protective of skeletal muscle contractility after an I/R injury. The results presented may have profound effects on future therapeutic recommendations and suggest possible pathways for clinical intervention to modulate I/R injury, which is commonplace in orthopaedic trauma and reconstructive surgery.

Functional electrical stimulation improves brain perfusion in cranial trauma patients.

OBJECTIVE: Demonstrate brain perfusion changes due to neuronal activation after functional electrical stimulation (FES). METHOD: It was studied 14 patients with hemiplegia who were submitted to a program with FES during fourteen weeks. Brain perfusion SPECT was performed before and after FES therapy. These patients were further separated into 2 groups according to the hemiplegia cause: cranial trauma and major vascular insults. All SPECT images were analyzed using SPM. RESULTS: There was a significant statistical difference between the two groups related to patient's ages and extent of hypoperfusion in the SPECT. Patients with cranial trauma had a reduction in the hypoperfused area and patients with major vascular insult had an increase in the hypoperfused area after FES therapy. CONCLUSION: FES therapy can result in brain perfusion improvement in patients with brain lesions due to cranial trauma but probably not in patients with major vascular insults with large infarct area.

Functional electrical stimulation improves brain perfusion in cranial trauma patients / A terapia com estímulo elétrico funcional pode melhorar a perfusão cerebral no SPECT em pacientes com trauma craniano

OBJECTIVE: Demonstrate brain perfusion changes due to neuronal activation after functional electrical stimulation (FES). METHOD: It was studied 14 patients with hemiplegia who were submitted to a program with FES during fourteen weeks. Brain perfusion SPECT was performed before and after FES therapy. These patients were further separated into 2 groups according to the hemiplegia cause: cranial trauma and major vascular insults. All SPECT images were analyzed using SPM. RESULTS: There was a significant statistical difference between the two groups related to patient's ages and extent of hypoperfusion in the SPECT. Patients with cranial trauma had a reduction in the hypoperfused area and patients with major vascular insult had an increase in the hypoperfused area after FES therapy. CONCLUSION: FES therapy can result in brain perfusion improvement in patients with brain lesions due to cranial trauma but probably not in patients with major vascular insults with large infarct area.

OBJETIVO: Demonstrar mudanças na perfusão cerebral devido à ativação neuronal depois de estimulação elétrica funcional (EEF). MÉTODO: Foram estudados 14 pacientes com hemiplegia submetidos a quatorze semanas de um programa com EEF. O SPECT de perfusão cerebral foi realizado antes e depois da terapia com EEF. Estes pacientes foram separados em 2 grupos com relação à causa da hemiplegia: trauma craniano e acidente vascular cerebral (AVC). As imagens de SPECT foram analisadas usando SPM. RESULTADOS: Houve diferença estatisticamente significativa entre os dois grupos relacionada a idade dos pacientes e extensão da hipoperfusão. Os pacientes com trauma craniano tiveram redução na área de hipoperfusão e pacientes com AVC tiveram aumento na área de hipoperfusão após terapia com EEF. CONCLUSÃO: A terapia com EEF pode levar a melhora na perfusão cerebral em pacientes com lesões cerebrais secundárias a trauma craniano; entretanto, provavelmente não em pacientes com extensas áreas de infarto secundárias a AVC.

Characterizing photothrombotic distal middle cerebral artery occlusion and YAG laser-induced reperfusion model in the Izumo strain of spontaneously hypertensive rats.

No study has systematically studied the relevance of original Izumo strain of spontaneously hypertensive rats (SHR/Izm) as a stroke model. Furthermore, both SHR/Izm and stroke-prone SHR/Izm (SHRSP/Izm) are commercially available, and recent progress in genetic studies allowed us to use several congenic strains of rats constructed with SHR/Izm and SHRSP/Izm as the genetic background strains. A total of 166 male SHR/Izm and 17 male SHRSP/Izm were subjected to photothrombotic middle cerebral artery (MCA) occlusion with or without YAG laser-induced reperfusion. The pattern of distal MCA was recorded. Infarct volumes were determined with 2,3,5-triphenyltetrazolium chloride. At 24 or 48 h after MCA occlusion, infarct volumes in the permanent occlusion and 2-h occlusion groups (88 ± 22 [SD] and 87 ± 25 mm³, respectively) were significantly larger than that in the 1-h occlusion group (45 ± 14 mm³), indicating the presence of sizeable zone of penumbra. Infarct size in SHRSP/Izm determined at 24 h after MCA occlusion was fairly large (124.0 ± 34.8 mm³, n = 10). Infarct volume in SHR/Izm with simple distal MCA was 76 ± 19 mm³, which was significantly smaller than 95 ± 22 mm³ in the other SHR/Izm with more branching MCA. These data suggest that this stroke model in SHR/Izm is useful in the preclinical testing of stroke therapies and elucidating the pathophysiology of cerebral ischemia/reperfusion.

Prevenção do estresse oxidativo na síndrome de isquemia e reperfusão renal em ratos com suplementação nutricional com antioxidantes / Prevention of oxidative stress in renal ischemia-reperfusion syndrome in rats with nutritional antioxidant supplementation

OBJETIVO: Verificar o potencial efeito protetor de suplementação com vitaminas antioxidantes em um modelo de síndrome de isquemia-reperfusão renal em ratos. MÉTODOS: Foram utilizados 29 ratos Wistar, divididos em três grupos: Grupo I e II (n=10 cada), submetidos a indução do estresse oxidativo pela aplicação de 60 minutos de isquemia renal, seguidos de 10 minutos de reperfusão; adicionalmente, os animais do Grupo II foram pré-tratados por doze dias com vitaminas antioxidantes (vitamina C 11,43mg/kg e vitamina E 28,57mg/kg) antes da submissão à isquemia; Grupo III (n=9), correspondendo aos animais Sham, que foram manipulados de forma equivalente aos outros grupos, porém sem indução do estresse oxidativo e sem suplementação antioxidante. Findo isso, as amostras de sangue e os rins foram colhidos para avaliação dos níveis do malondialdeído, do ácido úrico e da capacidade antioxidante total. RESULTADOS: Para o malondialdeído e ácido úrico do Grupo I foi observado um aumento estatisticamente significante (p<0,01) em relação ao Grupo III, o qual não apresentou diferença em relação ao Grupo II. Para os níveis de capacidade antioxidante total, foi encontrada uma diminuição nos animais do Grupo I em relação aos Grupos II e III (p<0,01). CONCLUSÃO: Esses dados confirmam não apenas a efetiva participação do estresse oxidativo neste modelo de síndrome de isquemia e reperfusão renal em ratos, como também que o uso de vitaminas antioxidantes, associadas à dieta, pode proteger os animais das alterações oxidativas.

OBJECTIVE: The objective of this study was to verify the potential protective effect of antioxidant vitamin supplementation in a model of renal ischemic-reperfusion injury in rats. METHODS: Twenty-nine Wistar rats were divided into three groups: groups I and II (n=10 each), were submitted to 60 minutes of renal ischemia, followed by 10 minutes of reperfusion; additionally, animals of group II were treated for twelve days with antioxidant vitamins (vitamin C 11.43mg/kg and vitamin E 28.5 mg/kg) before being submitted to ischemia; In group III (n=9), the animals were treated like the other groups but not submitted to ischemic-reperfusion injury and not given antioxidant supplements. Subsequently, blood samples and the kidneys were collected for assessment of malondialdehyde, uric acid and total antioxidant capacity. RESULTS: The malondialdehyde and uric acid of group I was significantly higher than those of group III (p<0.01), which in turn did not differ from group II. The levels of total antioxidant capacity of the animals of group I was lower than those of groups II and III (p<0.01). CONCLUSION: These data confirmed the effective participation of oxidative stress in this model of renal ischemia-reperfusion syndrome in rats and showed that the use of antioxidant supplementation can protect the animals from oxidative changes.

Reperfusion-induced temporary appearance of therapeutic window in penumbra after 2 h of photothrombotic middle cerebral artery occlusion in rats.

To explore the effects of reperfusion on evolution of focal ischemic injury, spontaneously hypertensive male rats were subjected to photothrombotic distal middle cerebral artery occlusion (MCAO) with or without YAG laser-induced reperfusion. The volume of fodrin breakdown zone, water content, and brain tissue levels of sodium (Na(+)) and potassium (K(+)) were measured in the ischemic core and penumbra. Reperfusion attenuated fodrin breakdown, and the volume containing fodrin breakdown product at 3 h after reperfusion (5 h after MCAO) (30+/-7 mm(3)) was significantly smaller than the 42+/-3 mm(3) of the permanent occlusion group. After 3 to 6 h of ischemia, Na(+) increased, and K(+) decreased in the ischemic core. Reperfusion after 2 h of MCA occlusion did not mitigate the ischemia-induced changes in brain tissue electrolytes and water content at 3 to 6 h of ischemia. Even in reperfusion after comparatively long periods of occlusion where brain infarction size, assessed 3 days after MCAO, was not significantly reduced by reperfusion, and the precipitating indicators of the ischemic core (Na(+), K(+), water content) did not improve, temporary improvement or a delay in progression of ischemic injury was discernible in the penumbra. These results indicate the possibility that treatment with reperfusion is permissive to the effects of neuroprotection.

A double blind, single centre, sub-chronic reperfusion trial evaluating FX06 following haemorrhagic shock in pigs.

OBJECTIVE: Haemorrhagic shock causes ischaemia and subsequent fluid resuscitation causes reperfusion injury, jointly resulting in high morbidity and mortality. We tested whether the anti-inflammatory fibrin-derived peptide, Bbeta(15-42), also called FX06, is tissue protective in a model of haemorrhagic shock. METHODS: In a pig model, we standardised the severity of haemorrhagic shock by achieving a cumulative oxygen deficit of approximately 100ml/kg body weight by withdrawing blood over a period of 1h. This was followed by resuscitation with shed blood and full electrolyte solution, and pigs were monitored for 3 days. At reperfusion, 17 pigs were randomly assigned to FX06 or solvent treatment. RESULTS: FX06-treated pigs demonstrated improved cardiac function (stroke volume index: 67ml/m(2) versus 33ml/m(2)), decreased troponin T release in the early reperfusion (0.24ng/ml versus 0.78ng/ml), decreased AST levels after 24h (106U/l versus 189U/l) and decreased creatinine levels after 24h (108micromol/l versus 159micromol/l). Furthermore, FX06-treated pigs demonstrated preservation of the gut/blood barrier, while controls demonstrated high endotoxin plasma levels indicating translocation of bacteria and/or its products (0.2EU/ml versus 24.3EU/ml) after 24h. This study also demonstrates a significantly improved neurological performance in the FX06 group as determined by S100beta serum levels (0.72microg/l versus 1.25microg/l) after 48h and neurological deficit scores (11 versus 70) after 24h. CONCLUSION: FX06 - when administered as an adjunct to fluid resuscitation therapy - is organ protective in pigs. Further investigations are warranted to reveal the protective mechanism of FX06.

Experimental renal preservation by normothermic resuscitation perfusion with autologous blood.

BACKGROUND: Normothermic perfusion (NP) has the potential to improve metabolic support and maintain the viability of ischaemically damaged organs. This study investigated the effects of NP compared with current methods of organ preservation in a model of controlled non-heart-beating donor (NHBD) kidneys. METHODS: Porcine kidneys (n = 6 in each group) were subjected to 10 min warm ischaemia and then preserved as follows: 2 h cold storage (CS) in ice (CS2 group), 18 h CS (CS18 group), 18 h cold machine perfusion (CP group) or 16 h CS + 2 h NP (NP group). Renal haemodynamics and function were measured during 3 h reperfusion with autologous blood using an isolated organ perfusion system. RESULTS: Increasing CS from 2 to 18 h reduced renal blood flow (mean(s.d.) area under the curve (AUC) 444(57) versus 325(70) ml per 100 g; P = 0.004), but this was restored by NP (563(119) ml per 100 g; P = 0.035 versus CS18). Renal function was also better in CS2, CP and NP groups than in the CS18 group (mean(s.d.) serum creatinine fall 92(6), 79(9) and 64(17) versus 44(13) per cent respectively; P = 0.001). The AUC for serum creatinine was significantly lower with CS for 2 h than for 18 h (mean(s.d.) 1102(2600) versus 2156(401) micromol/l.h; P = 0.001), although values in CP and NP groups were not significantly different from those in the CS2 group (1354(300) and 1756(280) micromol/l.h respectively). Two hours of NP increased the adenosine 3'-triphosphate : adenosine 3'-diphosphate ratio to a significantly higher level than the preperfusion values in all other groups (P = 0.046). CONCLUSION: NP with oxygenated blood was able to restore depleted ATP levels and reverse some of the deleterious effects of CS.

A novel way of liver preservation improves rat liver viability upon reperfusion.

BACKGROUND/AIM: Currently, the liver is cold-preserved at 0 approximately 4 degrees C for experimental and clinical purposes. Here, we investigated whether milder hypothermia during the initial phase of the preservation period was beneficial for liver viability upon reperfusion. METHODS: In the first set of experiments, rat livers were preserved either conventionally in clinically used histidine-trypthopan-ketoglutarate (HTK) solution (Group A: 45 min and Group B: 24 h) or by slow cooling HTK solution (from 13 degrees C to 3 degrees C) during the initial 45 min of preservation (Group C: 24 h). In the second set of experiments, additional groups of livers were evaluated: Group BB--preservation according to Group B and Group CC--preservation according to Group C. Further, some livers were preserved at 13 degrees C for 24 h. Livers were then reperfused using a blood-free perfusion model. RESULTS: Bile production was approximately 2-fold greater in Group C compared to Group B. Alanine transaminase (ALT) and aspartate transaminase (AST) release into perfusate were 2 approximately 3-fold higher in Group B compared to Group C. No significant differences were found in ALT and AST release between Group C and Group A. Livers in Group CC compared to Group BB exhibited significantly lower portal resistance, greater oxygen consumption and bromosulfophthalein excretion into bile and lower lactate dehydrogenase (LDH) release into perfusate. Histological evaluation of tissue sections in Group BB showed parenchymal dystrophy of hepatocytes, while dystrophy of hepatocytes was absent in Group CC. Livers preserved at 13 degrees C for 24 h exhibited severe ischemic injury. CONCLUSION: These results suggest that the conventional way of liver preservation is not suitable at least for rat livers and that slow cooling of HTK solution during the initial phase of cold storage can improve liver viability during reperfusion.

S-100B release during carotid endarterectomy under local anesthesia.

The neuronal protein S-100B has been found to be an indicator of cellular brain damage. The aim of the study was to evaluate whether cross-clamping of the carotid artery for carotid endarterectomy (CEA) under local anesthesia is associated with the same S-100B release pattern as during general anesthesia, where an increase in S-100B concentration in the jugular vein blood of 120% has been reported. In 45 consecutive patients undergoing CEA under local anesthesia, serum S-100B samples were drawn before surgery (T1), before carotid cross-clamping (T2), before cerebral reperfusion (T3), after reperfusion but before the end of surgery (T4), and 6 hr postoperatively (T5). At T1 and T5, blood samples were drawn only from the radial artery. Intraoperatively (T2-T4), samples were collected from the internal jugular vein additionally. S-100B levels were determined using an immunoluminometric assay (LIAISON) Sangtec 100; Sangtec, Bromma, Sweden). In eight patients, it was necessary to insert an intraluminal shunt because of signs of cerebral ischemia. In the remaining 37 patients, median carotid clamping time was 40 min. There were no neurological complications. There were no differences in baseline S-100B levels regarding gender and symptomatology. Median baseline (T1) and postoperative (T5) S-100B levels were identical (0.077 microg/L). All blood samples from the jugular vein showed significantly higher median S-100B levels than the corresponding arterial blood samples. Only slight increases of 13% and 18% were found during cross-clamping (T3) compared to the first intraoperative measurement (T2) in the venous and arterial samples, respectively, which was followed by decreases of 5% and 18%, respectively (T3-T4). S-100B release did not differ at any time point between patients who needed and patients who did not need a shunt, in either the arterial or the venous blood samples. During uncomplicated CEA under local anesthesia, there is no relevant increase of S-100B. These results are different from those reported when CEA is done under general anesthesia.

Neuroprotective role of Z-ligustilide against forebrain ischemic injury in ICR mice.

Radix Angelica sinensis, known as Danggui in Chinese, has been used to treat cardiovascular and cerebrovascular diseases in Traditional Chinese Medicine for a long time. Modern phytochemical studies showed that Z-ligustilide (LIG) is the main lipophilic component of Danggui. In this study, we examined whether LIG could protect ischemia/reperfusion-induced brain injury by minimizing oxidative stress and anti-apoptosis. Transient forebrain cerebral ischemia (FCI) was induced by the bilateral common carotid arteries occlusion for 30 min. LIG was intraperitoneally injected to ICR mice at the beginning of reperfusion. As determined via 2,3,5-triphenyl tetrazolium chloride (TTC) staining at 24 h following ischemia, the infarction volume in the FCI mice treated without LIG (22.1 +/- 2.6%) was significantly higher than that in the FCI mice treated with 5 mg/kg (11.8 +/- 5.2%) and 20 mg/kg (2.60 +/- 1.5%) LIG (P < 0.05 or P < 0.01). LIG treatment significantly decreased the level of malondialdehyde (MDA) and increased the activities of the antioxidant enzyme glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) in the ischemic brain tissues (P < 0.05 or P < 0.01 vs. FCI group). In addition, LIG provided a great increase in Bcl-2 expression as well as a significant decrease in Bax and caspase-3 immunoreactivities in the ischemic cortex. The findings demonstrated that LIG could significantly protect the brain from damage induced by transient forebrain cerebral ischemia. The antioxidant and anti-apoptotic properties of LIG may contribute to the neuroprotective potential of LIG in cerebral ischemic damage.