Rabbit Brain Research Articles

Histopathological effect of energy drinks (Red Bull) on Brain, Liver, Kidney, and Heart in Rabbits

Background: Red bull has been known as a healthy drink within many populations. The chemical composition of energy drinks can produce multiple adverse effects, including serious behavioral effects. Objective: To study the effect of energy drinks (Red Bull) on brain, liver, kidney, and heart in rabbits. Methods: thirty males albino rabbits were used, animals divided into 3 groups (A, B, and C), each group contain 10 rabbits. Group A treated with high dose of red bull (10 cc), Group B treated with low dose (5 cc), and Group C is control group. Results: The results showed that control group had no any pathological changes while both Groups A and B showed many pathological changes, Group A showed renal vascular congestion, hemorrhage of interstitial tissue, focal atrophy, and degeneration of lining epithelium of proximal and distal convoluted tubules, while Group B showed renal vascular congestion, glomerular capillary congestion, hemorrhage of interstitial tissue with swelling of lining epithelium of proximal and distal convoluted tubules that appeared as pale cytoplasm with star-shaped lumen, and swelling of glomeruli; this reveals that there is dose–response relationship between treated groups with low and high dose of red bull. Conclusion: Energy drinks have dose–response relationship with adverse effect; it has become apparent that the consumption of these energy drinks will seriously harm the body.

Trajectory of inflammatory and microglial activation markers in the postnatal rabbit brain following intrauterine endotoxin exposure

BackgroundMaternal infection is a risk factor for periventricular leukomalacia and cerebral palsy (CP) in neonates. We have previously demonstrated hypomyelination and motor deficits in newborn rabbits, as seen in patients with cerebral palsy, following maternal intrauterine endotoxin administration. This was associated with increased microglial activation, primarily involving the periventricular region (PVR). In this study we hypothesized that maternal intrauterine inflammation leads to a pro-inflammatory environment in the PVR that is associated with microglial activation in the first 2 postnatal weeks. MethodsTimed pregnant New Zealand white rabbits underwent laparotomy on gestational day 28 (G28). They were randomly divided to receive lipopolysaccharide (LPS; 20μg/kg in 1mL saline) (Endotoxin group) or saline (1mL) (control saline, CS group), administrated along the wall of the uterus. The PVR from the CS and Endotoxin kits were harvested at G29 (1day post-injury), postnatal day1 (PND1, 3day post-injury) and PND5 (7days post-injury) for real-time PCR, ELISA and immunohistochemistry. Kits from CS and Endotoxin groups underwent longitudinal MicroPET imaging, with [11C]PK11195, a tracer for microglial activation. ResultsWe found that intrauterine endotoxin exposure resulted in pro-inflammatory microglial activation in the PVR of rabbits in the first postnatal week. This was evidenced by increased TSPO (translocator protein) expression co-localized with microglia/macrophages in the PVR, and changes in the microglial morphology (ameboid soma and retracted processes). In addition, CD11b level significantly increased with a concomitant decline in the CD45 level in the PVR at G29 and PND1. There was a significant elevation of pro-inflammatory cytokines and iNOS, and decreased anti-inflammatory markers in the Endotoxin kits at G29, PND1 and PND5. Increased [11C]PK11195 binding to the TSPO measured in vivo by PET imaging in the brain of Endotoxin kits was present up to PND14–17. ConclusionsOur results indicate that a robust pro-inflammatory microglial phenotype/brain milieu commenced within 24h after LPS exposure and persisted through PND5 and in vivo TSPO binding was found at PND14–17. This suggests that there may be a window of opportunity to treat after birth. Therapies aimed at inducing an anti-inflammatory phenotype in microglia might promote recovery in maternal inflammation induced neonatal brain injury.

Quantitative analysis of TNF-α, IL-4, and IL-10 expression, nitric oxide response, and apoptosis in Encephalitozoon cuniculi–infected rabbits

The expression of tumor necrosis factor (TNF) -α, interleukin (IL) −4 and IL-10, as well as apoptosis and nitric oxide (NO) levels were measured in the brain and kidneys of immunocompetent and immunosuppressed New Zealand White rabbits infected with Encephalitozoon cuniculi. All of the animals had clinical signs histopathological lesions compatible with encephalitozoonosis and were E. cuniculi-positive by using a carbon immunoassay test. Encephalitozoon cuniculi infection promoted the expression of TNF-α and NO production in the kidneys of infected rabbits, and a synergic effect was observed in animal treated with dexamethasone. The IL-4 expression was similar in the brain and kidneys of infected rabbits, regardless of their immunologic status. The IL-10 mRNA expression in the brain of infected immunosuppressed rabbits was elevated when compared with positive controls. Apoptosis of granuloma mononuclear-like cells was detected in immunocompetent E. cuniculi–infected rabbits, but it was more evident in infected-immunosuppressed animals. Nitric oxide levels were elevated both in immunocompetent and immunosuppressed infected animals, but it was more apparent in the kidneys. These data suggest that modulation of the immune response by E. cuniculi could contribute to the survival of the parasite within phagocytic cells in granulomas via an as yet undetermined mechanism.

Oxidative stress prediction: A preliminary approach using a response surface based technique

A response surface was built to predict the lipid peroxidation level, generated in an iron-ascorbate in vitro model, of any organ, which is correlated with the oxidative stress injury in biological membranes. Oxidative stress studies are numerous, usually performed on laboratory animals. However, ethical concerns require validated methods to reduce the use of laboratory animals. The response surface described here is a validated method to replace animals. Tissue samples of rabbit liver, kidney, heart, skeletal muscle and brain were oxidized with different concentrations of FeCl3 (0.1 to 8mM) and ascorbate (0.1mM), during different periods of time (0 to 90min) at 37°C. Experimental data obtained, with lipid content and antioxidant activity of each organ, allowed constructing a multidimensional surface capable of predicting, by interpolation, the lipid peroxidation level of any organ defined by its antioxidant activity and fat content, when exposed to different oxidant conditions. To check the predictive potential of the technique, two more experiments were carried out. First, in vitro oxidation data from lung tissue were collected. Second, the antioxidant capacity of kidney homogenates was modified by adding melatonin. Then, the response surface generated could predict lipid peroxidation levels produced in these new situations. The potential of this technique could be reinforced using collaborative databases to reduce the number of animals in experimental procedures.

Efficacy of α-lipoic acid against cadmium toxicity on metal ion and oxidative imbalance, and expression of metallothionein and antioxidant genes in rabbit brain.

To explore the protective efficacy of α-lipoic acid (ALA) against Cd-prompted neurotoxicity, young male New Zealand rabbits (Oryctolagus cuniculus) were divided randomly into four groups. Group 1 (control) received demineralized water. Group 2 (Cd) administered cadmium chloride (CdCl2) 3mg/kg bwt. Group 3 (ALA) administered ALA 100mg/kg bwt. Group 4 (Cd+ALA) administered ALA 1h after Cd. The treatments were administered orally for 30 consecutive days. Cd-induced marked disturbances in neurochemical parameters were indicated by the reduction in micro- and macro-elements (Zn, Fe, Cu, P, and Ca), with the highest reduction in Cd-exposed rabbits, followed by Cd+ALA group and then ALA group. In the brain tissues, Cd has significantly augmented the lipid hydroperoxides (LPO) and reduced the glutathione (GSH) and total antioxidant capacity (TAC), and glutathione peroxidase and glutathione S-transferase enzyme activities but had an insignificant effect on the antioxidant redox enzymes. Administration of ALA effectively restored LPO and sustained GSH and TAC contents. Moreover, Cd downregulated the transcriptional levels of Nrf2, MT3, and SOD1 genes, and upregulated that of Keap1 gene. ALA treatment, shortly following Cd exposure, downregulated Keap1, and upregulated Nrf2 and GPx1, while maintained MT3 and SOD1 mRNA gene expression in the rabbits' brain. These data indicated the ALA effectiveness in protecting against Cd-induced oxidative stress and the depletion of cellular antioxidants in the brain of rabbits perhaps due to its antioxidant, free radical scavenging, and chelating properties.

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OBJECTIVES/SPECIFIC AIMS: To explore the racial differences in rotational thromboelastometry findings using whole blood and plasma samples from healthy volunteers. METHODS/STUDY POPULATION: We studied a cohort of patients at Tulane University Hospitals who came into the pre-op clinic to get blood drawn for labs. The cohort included a total of 44 patients who were otherwise healthy adult volunteers with no history of cardiovascular nor thromboembolic events, 30 African Americans and 14 Caucasians. Patients who required lab work for their upcoming surgery were asked to participate in the study by giving a sample of blood collecting in a light blue-top sodium citrate tube. We excluded patients who were currently on any anticoagulation or antiplatelet medications. We also excluded those with current or previous history of cancer, those with known bleeding disorder, and those who were on chronic transfusion protocol, or had received a blood transfusion within the last 21 days. Data collection was carried out after informed consent was obtained; we collected citrated whole-blood (WB) samples. WB samples were processed within 3 hours of phlebotomy. Platelet free plasma, obtained after centrifugation at 2500 cGy of whole blood for 20 minutes, was kept frozen at −70°C. Frozen plasma was thawed at 37°C for 5 minutes before testing. Samples were recalcified with star-tem reagent, and then the in-tem reagent was added. The latter contains an optimized concentration of ellagic acid and partial thromboplastin phospholipid from rabbit brain. Thromboelastometry (ROTEM) parameters including clotting time, clot formation time, alpha angle, maximum clot firmness, and Lysis Index after 30 and 45 minutes were determined. Data was then retrieved from the ROTEM database and put into an Excel sheet to be analyzed. RESULTS/ANTICIPATED RESULTS: Our results showed that the CFT was higher in both the plasma and the WB of Caucasians when compared with African Americans with a difference between means 137.5±233.7 (p=0.56) and 11±7.85 (p=0.168), respectively; while MCF was increased in the WB and plasma of AA with a difference between means of 1.719±1.974 (p=0.38) and 5.37±2.49 (p=0.037), respectively. In other words, the plasma of Caucasians did seem to take longer to reach the maximum firmness (however not statistically significant p>0.05), while the maximum clot firmness was significantly higher in plasma of AA. In summary and compatibly with the previously published data, our results showed significantly increased prothrombotic profile in the plasma of African Americans when compared with Caucasians. DISCUSSION/SIGNIFICANCE OF IMPACT: This reinforces the role of the whole vascular system and the interaction between its different components in the pathophysiology of thromboembolic events. In one case control study, African ethnicity was associated with increased risk of DVT in parallel with significantly increased peak thrombin on thrombin generation when compared with Caucasians. With our preliminary results, we confirm these data using another tool for the assessment of the plasma in addition to comparing WB samples too. More prospective studies, with higher number of subjects evaluating the value of the results in predicting the risk of development of thromboembolic events in different ethnicities, are needed for better understanding of this disease. In addition, thromboelastometry might require adjustment for ethnicity in studies evaluating ethnically diverse populations.

POCT PT INR — Is it adequate for patient care? A comparison of the Roche Coaguchek XS vs. Stago Star vs. Siemens BCS in patients routinely seen in an anticoagulation clinic

BackgroundIn this study we examined the difference in patient INR values as measured by the POCT CoaguChek XS device and central laboratory Stago Evolution and Siemens BCS XP analyzers. MethodsThis study composed of 100 warfarin therapy patients and 20 coagulation normal subjects, showed that the difference between the POCT and clinical laboratory values increased with increasing INR and was exacerbated by the use of different thromboplastin reagents by the POCT and central lab. ResultsThe CoaguChek XS and on-site Stago analyzers which used human recombinant (ISI=1.01) and rabbit brain thromboplastin (ISI=1.25), respectively, showed reasonable agreement for INR<3.0 (k=0.62) but significant difference for INR≥3.0 (k=0.10). In contrast, the CoaguChek XS and Siemens BCS XP, which both employed human recombinant thromboplastin (BCS ISI=1.02), showed greater agreement for the complete range INR values (INR<3.0 k=0.84; INR≥3.0 k=0.70). ECAA Poller calibrant data showed the automated instruments were performing as expected, indicating that ISI calibrations were correct but insufficient to standardize the INR values for the different thromboplastin methods across the full range of measured INRs. Central lab verification of POCT INR>5.0 with the Stago Evolution prevented adverse treatment events for the warfarin therapy patients in the six months preceding and following this investigation.

Recombinant Lactobacillus casei expressing Clostridium perfringens toxoids α, β2, ε and β1 gives protection against Clostridium perfringens in rabbits

The present study used Lactobacillus casei ATCC 393 as antigen delivery system to express C. perfringens toxoids α-β2-ε-β1 to construct the recombination Lactobacillus casei pPG-2-α-β2-ε-β1/L. casei 393. After being induced by 1% xylose, the specificity and integrity of recombinant strain were determined by Western-blotting. Rabbits as native animal model were immunized orally with pPG-2-α-β2-ε-β1/L. casei 393 and the titers of specific IgG and sIgA were determined by ELISA. The result showed that oral administration with the recombinants could elicit both local mucosal and systemic immune responses. The proliferation of spleen lymphocytes in rabbits immunized with pPG-2-α-β2-ε-β1/L. casei 393 was observed. Levels of IL-4 and IFN-γ produced were significantly higher in lymphocytes isolated from the vaccine group than those from the control groups. Flow cytometry assay showed that both the percentages of CD4+T cells and CD8+T cells from the vaccine group were significantly increased than the control groups. All these results showed that immunizing with recombinants can elicit both humoral immunity and cellular immunity. Besides, in order to determine the effectiveness of oral immunization with pPG-2-α-β2-ε-β1/L. casei 393, rabbits of vaccine group and control groups were challenged with 1×LD100 unit of culture filtrate of C. perfringens type C and type D toxins respectively. After challenge, 100% of the immunized rabbits survived, while the rabbits of the control group were killed within 48h. Observation on histopathology showed that histopathological changes were obviously found in heart, liver, spleen, lung, kidney, intestine and brain of rabbits from the control groups, while no apparent histopathological change was observed in the vaccine group. All the results show that pPG-2-α-β2-ε-β1/L. casei 393 can eliciteffective immunoprotection against C. perfringens. All of these suggest that the use of pPG-2-α-β2-ε-β1/L. casei 393 can be regarded as candidate for the development of a vaccine against C. perfringens.

Magnetic inductive phase shift: a new method to differentiate hemorrhagic stroke from ischemic stroke on rabbit

BackgroundThe major therapy for ischemic stroke is thrombolytic treatment, but severe consequences occur when this method is used to treat hemorrhagic stroke. Currently, computed tomography and magnetic resonance imaging are used to differentiate between two types of stroke, but these two methods are ineffective for pre-hospital care.MethodsWe developed a new brain diagnostic device for rabbits based on electromagnetic induction to non-invasively differentiate two types of stroke. The device includes two coils and a phase difference measurement system that detects the magnetic inductive phase shift (MIPS) value to reflect the tissue’s condition. The hemorrhage model was established through the injection of autologous blood into the internal capsule of a rabbit’s brain. Ischemia was induced in the brain of a rabbit by bilateral carotid artery occlusion. Two types of animal models were measured with our device.ResultsThe MIPS value gradually decreased with increasing injected blood and increased with ischemia time. The MIPS changes induced by the two types of strokes were exact opposites, and the absolute values of MIPS variation in the hemorrhagic and the ischemic groups were significantly larger than those of the normal control group (P < 0.05).ConclusionsThe tested technique can differentiate ischemic stroke from hemorrhagic stroke on rabbit brain in a non-invasive, continuous, and bulk monitoring manner by using a simple and inexpensive apparatus.

The protective effect of PI3K/AKt/GSK3β signaling pathway on brain during resuscitation with neck cooling

Objective To study the changes of PI3K/Akt/GSK3β signaling pathway during resuscitation with neck cooling in order to explore the relationship between the protective effect of neck cooling and the phosphorylation of PI3K/Akt and GSK3β. Methods Thirty rabbits were randomly(random number) divided into five groups, and models of cadiac arrest were induced by ventricular fibrillation(VF, the positive electrode in the right ventricle and negative pole on the apex of heart) for 4 min. In sham group, a electrode was placed into right ventricle without electric current conducted, and CA was not induced. The rabbits were sacrificed and specimens were taken at 24 hours after modeling. In normothermia treat group(NT group), resuscitation was carried out to restoration of spontaneous circulation(ROSC), and the rabbits were sacrificed and specimens were taken at 24 hours after modeling. In intra-arrest therapeutic hypothermia group (IATH group), rapid neck cooling was initiated at the same time with CPR, and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC. Rabbits were sacrificed and specimens were taken at 24 hours after modeling. In recovery period cooling + LY294002 group(PATH+ LY294002 group), LY294002 was injected intra-ventricularly at 20 minutes before resuscitation. Rapid neck cooling was started at the same time with CPR, and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC. The rabbits were sacrificed and specimens were taken at 24 hours after modeling. In post-arrest therapeutic hypothermia group (PATH group), rapid neck cooling was begun after CPR for 1 hour, and the target brain temperature was set at 34 ℃ maintained for 4 hours after ROSC. The rabbits were sacrificed and specimens were taken at 24 hours after modeling. Animals were sacrificed by using overdose anesthetic drug. Western blot was used to detect the level of Akt p-Akt GSK-3β p-GSK-3β (ser9) protein, and TUNEL was used to observe apoptosis of tissues in each group. Multiple comparisons were performed with one-way analysis of variance (ANOVA). Results Compared with Sham group, Akt (Thr-308) phosphorylation (P-AKT) and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in NT group was significantly reduced, and showed a gradual reduction trend (P<0.05); the P-AKT and P-GSK-3β levels in the brain neuron cytoplasm in 24 hours after CPR resuscitation in IATH group were significantly enhanced compared with NT group (P<0.05); the levels of these two kinds of protein at one hour after resuscitation in PATH group were significantly enhanced compared with NT group (P<0.05), but lower in IATH group. Intra-ventricularly injection of LY294002 made the effect of hypothermia lost, indicating that LY294002 inhibited the phosphorylation of Akt. Apoptosis cells were significantly reduced in IATH group and normothermia theatment group compared with PATH group and LY294002 group(P<0.05). Conclusions Neck cooling can reduce apoptosis in rabbit brain cells after recovery, and the protective effect on brain is best in intra-arrest therapeutic hypothermia group. LY294002 specifically block the PI3K/Akt pathway, and the protective effect of cooling on the brain can be abolished, indicating hypothermia protects the neurological function via activation of PI3K/Akt pathway. Neck cooling protects the neurological function by activating PI3K/Akt/GSK-3β, promoting the Akt activation, and increasing the expression of P-GSK3β. Specific Akt inhibitor LY294002 inhibits Akt phosphorylation of brain tissue recovery and further inhibit the phosphorylation of GSK-3β, thus abolishing protective effect of cooling on neurological function. Key words: Cardiac arrest; Cardiopulmonary resuscitation; Hypothermia; PI3K/Akt/GSK3β; Rabbit

298 - Intra-arterial delivery of human mesenchymal stem cells into the brains of New Zealand white rabbits

298 - Intra-arterial delivery of human mesenchymal stem cells into the brains of New Zealand white rabbits

The neural pathway for lacrimal gland tear secretion in New Zealand White rabbits

ObjectiveWe investigated the neural pathway for tear secretion from the lacrimal gland of New Zealand White rabbits. MethodsNine healthy adult New Zealand White rabbits were randomly divided into three experimental groups, namely, an irritant-stimulated group, a non-stimulated group, and a saline-stimulated group. Sanitized dry cotton swabs with menthol were used to wipe both of the rabbits’ eyelids in the irritant-stimulated group, and the non-stimulated group and saline- stimulated group were compared as controls. The animals in the three groups were killed 2h later and the expressions of c-Fos in the frontal cortex, hippocampus, hypothalamus, pons, and medulla oblongata of the rabbits were detected using immunofluorescence labeling. According to the distribution of c-Fos protein expression, 12 healthy adult New Zealand rabbits were similarly divided into three groups for retrograde tract tracing via pseudorabies virus (PRV) injection into the lacrimal gland. Immunofluorescence labeling was used to analyze PRV-infected neurons in the brains of rabbits after survival for 30h, 38h, and 46h. ResultsThe most c-Fos-positive immunolabeled cells were observed in the menthol-stimulated group, whereas fewer c-Fos-positive immunolabeled cells were observed in the saline-stimulated group.The non-treated group showed the least c-Fos-positive immunolabeled cells. At 30h after PRV injection, PRV-positive neurons were found only in the superior salivary nucleus of the pons (SSN). At 38h, PRV-infected neurons were observed in the lateral nucleus of the superior olive (LSO) and the medial nucleus of the superior olive (MSO). At 46h, PRV-infected neurons were found in the nucleus of the trapezoid body (Tz) and the hypothalamic paraventricular nucleus (PVN), and their distributions were dense in the LSO and MSO. ConclusionsMenthol-induced c-Fos protein expression and PRV-mediated tract tracing suggest that in New Zealand White rabbits, the neural pathway that regulates tear secretion from the lacrimal gland proceeds from the PVN to the superior olivary complex of the pons to the SSN and finally to the lacrimal gland.

In Vivo Bioimpedance Spectroscopy Characterization of Healthy, Hemorrhagic and Ischemic Rabbit Brain within 10 Hz-1 MHz.

Acute stroke is a serious cerebrovascular disease and has been the second leading cause of death worldwide. Conventional diagnostic modalities for stroke, such as CT and MRI, may not be available in emergency settings. Hence, it is imperative to develop a portable tool to diagnose stroke in a timely manner. Since there are differences in impedance spectra between normal, hemorrhagic and ischemic brain tissues, multi-frequency electrical impedance tomography (MFEIT) shows great promise in detecting stroke. Measuring the impedance spectra of healthy, hemorrhagic and ischemic brain in vivo is crucial to the success of MFEIT. To our knowledge, no research has established hemorrhagic and ischemic brain models in the same animal and comprehensively measured the in vivo impedance spectra of healthy, hemorrhagic and ischemic brain within 10 Hz–1 MHz. In this study, the intracerebral hemorrhage and ischemic models were established in rabbits, and then the impedance spectra of healthy, hemorrhagic and ischemic brain were measured in vivo and compared. The results demonstrated that the impedance spectra differed significantly between healthy and stroke-affected brain (i.e., hemorrhagic or ischemic brain). Moreover, the rate of change in brain impedance following hemorrhagic and ischemic stroke with regard to frequency was distinct. These findings further validate the feasibility of using MFEIT to detect stroke and differentiate stroke types, and provide data supporting for future research.

Bioamine-crosslinked gellan gum hydrogel for neural tissue engineering

Neural tissue engineering and three-dimensional in vitro tissue modeling require the development of biomaterials that take into account the specified requirements of human neural cells and tissue. In this study, an alternative method of producing biomimetic hydrogels based on gellan gum (GG) was developed by replacing traditional crosslinking methods with the bioamines spermidine and spermine. These bioamines were proven to function as crosslinkers for GG hydrogel at +37 °C, allowing for the encapsulation of human neurons. We studied the mechanical and rheological properties of the formed hydrogels, which showed biomimicking properties comparable to naïve rabbit brain tissue under physiologically relevant stress and strain. Human pluripotent stem cell-derived neuronal cells demonstrated good cytocompatibility in the GG-based hydrogels. Moreover, functionalization of GG hydrogels with laminin resulted in cell type-specific behavior: neuronal cell maturation and neurite migration.

Safety of Intra-Arterial Injection with Tumor-Activated T Cells to the Rabbit Brain Evaluated by MRI and SPECT/CT

Glioblastoma multiforme (GBM) is the most common and most severe form of malignant gliomas. The prognosis is poor with current combinations of pharmaceutical, radiotherapy, and surgical therapy. A continuous search for new treatments has therefore been ongoing for many years. Therapy with tumor-infiltrating lymphocytes (TILs) is a clinically promising strategy to treat various cancers, including GBM. An endovascular intraarterial injection of TILs as a method of delivery may, instead of intravenous infusion, result in better retention of effector cells within the tumor. Prior to clinical trials of intra-arterial injections with any cells, preclinical safety data with special emphasis on embolic–ischemic events are necessary to obtain. We used native rabbits as a model for intra-arterial injections with routine clinical catheter material and a clinical angiography suite. We selectively infused a total dose of 20 million activated T cells at a cell concentration of 4,000 cells/ml over 8 min of injection time. The rabbits were evaluated for ischemic lesions by 9.4 T magnetic resonance imaging (MRI) (n = 6), and for tracking of injected cells, single-photon emission computed tomography/computed tomography (SPECT/CT) was used (n = 2). In this study, we show that we can selectively infuse activated T cells to a CNS volume of 3.5 cm3 (estimated from the volumetric MRI) without catastrophic embolic– ischemic adverse events. We had one adverse event with a limited basal ganglia infarction, probably due to catheter-induced mechanical occlusion of one of the lateral lenticulostriatal arteries. The cells pass through the native brain without leaving SPECT signals. The cells then, over the first hours, end up in the liver to a large extent and to a lesser degree by the spleen, pancreas, and kidneys. Virtually no uptake could be detected in the lungs. This indicates a difference in biodistribution as opposed to other cell types when infused intravenously.

Rabbit Model of Human Gliomas: Implications for Intra-Arterial Drug Delivery.

The prognosis for malignant brain tumors remains poor despite a combination of surgery, radiotherapy, and chemotherapy. This is partly due to the blood-brain barrier, a major obstacle that prevents therapeutic agents from effectively reaching the tumor. We have recently developed a method for precise and predictable opening of the blood-brain barrier via the intra-arterial administration of mannitol, a hyperosmolar agent, in a rabbit model, whose vascular anatomy facilitates the use of standard interventional neuroradiology techniques and devices. To date, however, no protocols are available that enable human glioma modeling in rabbits. In this article, we report on the xenotransplantation of a human glioblastoma (GBM-1) in adult New Zealand rabbits. We induced multi-drug immunosuppression (Mycophenolate Mofetil, Dexamethasone, Tacrolimus) and stereotactically implanted GBM-1 tumor cells into rabbit brains. The rabbits were followed for 42 days, monitored by MRI and body weight measurements, and underwent postmortem histopathological analysis. On MRI, brain tumors were identified on T2-weighted scans. On histopathology, tumors were detected with hematoxylin/eosin and their human origin was confirmed with immunohistochemistry against human-specific antigens. Our method for human glioma modeling in rabbits provides the foundation to test novel treatment strategies, including intra-arterial therapeutic agent delivery.

Effects of Acute Toluene Toxicity on Different Regions of Rabbit Brain.

The acute phase effects of toluene on the brain have been investigated in this study using rabbit brain via histopathological, immunohistochemical, and biochemical methods. A total of 20 male rabbits were used as control and experimental groups. Moreover, nerve growth factor (NGF), tumor necrosis factor-alpha (TNF-alpha), dopamine (DA), and glial fibrillary acidic protein (GFAP) tests were performed in order to designate the severity of the biochemical damage. In the biochemical evaluation of the prefrontal cortex, hippocampus, hypothalamus, substantia nigra, and entorhinal cortex, the TNF-alpha levels in the brain were found to be significantly higher than in the control group. Levels of dopamine, secreted from the substantia nigra, nerve growth factor (NGF) developed from the hippocampal neurons, and GFAP, secreted from astrocyte cells, were detected to be significantly lower in the toluene-administration group than in the control group (p < 0.05). In addition, areas of focal vacuolar degeneration (abscess formation), gliosis, and perivascular demyelination, many pyknotic cells and necrosis were observed. In the toluene-administration group compared to the control group, distinct excessive expansions of the blood vessels and severe degeneration in the structure of cells and also dispersed cell borders were observed. Furthermore, abnormal malformations of the nuclei structure of the oligodendrocyte cells were seen. Bodies of the sequential neurons of the hippocampus in the toluene-administration group were distinctly structurally damaged compared to the control group. In addition, cytoplasm of the cortex cell showed serious immune reactivity in the experimental group.

Identification and Characterization of a New Mutation in Coagulation Factor VII (Carmel mutation), with Different Reactivity Towards Tissue Factors of Different Origin

Inherited factor VII (FVII) deficiency is a rare autosomal recessive disorder manifested by bleeding tendency in homozygotes or compound heterozygotes. The bleeding tendency is variable and does not always correlate with the measured amount of FVII. More than 30 mutations causing FVII deficiency have been identified. The aim of this study was to identify the molecular defect in a family with FVII abnormality and to characterize the functional defect and its clinical consequences.An 80 year old woman showed various prolonged prothrombin time (PT) dependent on the origin of tissue factors (TF) in the reagents, in the range of 5-18%. FVII activity measured using placenta human TF was ~4% and less than 1% using recombinant human TF or rabbit brain TF. Direct sequencing of PCR-amplified FVII genomic DNA exons demonstrated a single homozygous nucleotide exchange G>A in exon 6 predicting missense mutation Tyr164Cys, in close vicinity to the active site of the protease and to the binding site of FX.Sequencing of exon 6 in six members of the family comprising three generations was performed. All of them exhibited low activity of FVII. Direct sequencing of PCR-amplified exon 6 demonstrated that her son, daughter and grand-daughter are heterozygote to the novel mutation, Tyr164Cys, henceforth, Carmel mutation. The grand -daughter (third generation) was identified as a compound heterozygote carrying in addition an Ala244Val mutation along with a Arg353Gln polymorphism, a well-known linkage previously characterized in the Israeli population, inherited from the paternal side. Her FVII activity was 3-7% and 8% FVII antigen. In the individuals heterozygote for Tyr164Cys we detected 41-54% activity and 46-66% antigenic FVII.The measurement of VIIa in the proband and her grand-daughter could not demonstrate any activity in plasma, possibly because of perturbation of the interaction between the mutated FVII and TF or FX which are part of the assay. The heterozygote for Tyr164Cys showed approximately 30% FVIIa of normal controls .Despite the dramatic laboratory presentation, the clinical manifestation of the Tyr164Cys genotype is very mild, suggesting that a low level of FVII is sufficient to prevent bleeding. The homozygous patient reached an old age, gave birth to two children and did not suffer any major bleedings during her life. The compound heterozygote presents a similar phenotype with slightly higher FVII activity and antigen, no measurable FVIIa activity and mild clinical presentation. Further investigation into this novel mutation is needed in order to characterize the interaction of the FVII with TF, phospholipids and FX and to understand the reason for such low amount of FVII protein in plasma. DisclosuresNo relevant conflicts of interest to declare.

Toxicity of Fe3O4 nanoparticles on oxidative stress status, stromal enzymes and mitochondrial respiration and swelling of Oryctolagus cuniculus brain cortex

Fe3O4 nanoparticles are the most widely used metal oxide nanoparticles especially, in biomedical applications. Although, nanoparticles can enter to the different organs, little is known so far on the neurotoxic potential and oxidative stress of Fe3O4. Here the understanding of the effect of Fe3O4 nanoparticles on the general Redox state of rabbit brain and the effect on mitochondrial swelling and respiration were assessed. Fe3O4 resulted in increase of brain markers, lipid peroxidation, protein and ROS formation. Mitochondrial enzymes and swelling were elevated with decreased respiration level. Caspase 3 activity and TNF-α level were also increased. Finally, our study suggested that the mitochondrial disease and dysfunction with elevated oxidative stress in rabbit brain treated with 200 and 300 μg/kg per Os is the original of neurotoxicity and maybe the original cause of neurodegenerative disease.

Ex-Vivo Characterization of Bioimpedance Spectroscopy of Normal, Ischemic and Hemorrhagic Rabbit Brain Tissue at Frequencies from 10 Hz to 1 MHz.

Stroke is a severe cerebrovascular disease and is the second greatest cause of death worldwide. Because diagnostic tools (CT and MRI) to detect acute stroke cannot be used until the patient reaches the hospital setting, a portable diagnostic tool is urgently needed. Because biological tissues have different impedance spectra under normal physiological conditions and different pathological states, multi-frequency electrical impedance tomography (MFEIT) can potentially detect stroke. Accurate impedance spectra of normal brain tissue (gray and white matter) and stroke lesions (ischemic and hemorrhagic tissue) are important elements when studying stroke detection with MFEIT. To our knowledge, no study has comprehensively measured the impedance spectra of normal brain tissue and stroke lesions for the whole frequency range of 1 MHz within as short as possible an ex vivo time and using the same animal model. In this study, we established intracerebral hemorrhage and ischemic models in rabbits, then measured and analyzed the impedance spectra of normal brain tissue and stroke lesions ex vivo within 15 min after animal death at 10 Hz to 1 MHz. The results showed that the impedance spectra of stroke lesions significantly differed from those of normal brain tissue; the ratio of change in impedance of ischemic and hemorrhagic tissue with regard to frequency was distinct; and tissue type could be discriminated according to its impedance spectra. These findings further confirm the feasibility of detecting stroke with MFEIT and provide data supporting further study of MFEIT to detect stroke.