It has been estimated that approximately 300,000 Canadians are afflicted with schizophrenia (SCZ). Due to the severity of symptoms as well as critical age of onset, the quality of life among SCZ patients can be poor; thus, further therapeutic research is of great interest. In addition to the more-common rodent models of SCZ (amphetamine sensitization, PCP sensitization, etc.), our lab has proposed the medial prefrontal cortex (mPFC) synapsin-II knockdown (KD) model. Prior to this study, the cognitive effects of mPFC synapsin-II KD had yet to be reported. Using a 14-day continuous infusion of antisense (AS) deoxyoligonucleotides for synapsin-II to the mPFC, the results of the KD model have been recreated for further study. The 5-choice-serial-reaction-time-task was utilized to determine the effects of mPFC synapsin-II KD on attention and vigilance. In addition to reductions in both PPI (p<0.05) and social interaction (p<0.05), as well as hyper-locomotion (p<0.05); rats treated with synapsin II AS performed significantly worse on the 5-CSRTT than did control (mismatch/aCSF) animals. The AS animals were significantly less likely to make correct responses (p<0.001), and significantly more likely to commit omissions (p<0.0001) and perseverative responses (p<0.05) than were control animals. Additionally, an [18F]FDG tracer and PET/CT scans were used to determine differences in brain metabolism due to synapsin-II KD. Results revealed significant reductions in [18F]FDG among AS treated rats (p<0.05) when compared to control animals. This reduction appeared to be a global result, however it followed 13-days of treatment which may account for the widespread effect. The mPFC synapsin-II KD model of SCZ has showcased cognitive and metabolic effects similar to that of SCZ. These findings, in conjunction with past research, provide evidence for the synapsin-II KD model as a viable rodent model of SCZ. Further research utilizing this model will provide valuable insight into the pathogenesis of SCZ and potential therapeutics. / Thesis / Master of Health Sciences (MSc)

Remote ischemic conditioning (RIC) is a noninvasive procedure where blood flow to a limb is repetitively reduced, sometimes called an “exercise memetic”. RIC delivered before (pre-RIC) or after (post-RIC) stroke is reportedly neuroprotective in preclinical stroke models. A review of the preclinical RIC literature revealed that studies almost exclusively use male subjects and a single stroke model (MCAO) that produces a large injury (~34% of hemisphere). To improve clinical translation, efficacy should be demonstrated in multiple stroke models and both sexes. Furthermore, the restorative potential of RIC (delivered past the neuroprotection window) to improve stroke recovery remains to be investigated. In male and female Sprague-Dawley rats (n=129) a standardized session (5min inflation, 5min deflation, 4 repetitions) of RIC was delivered using a pressurized cuff on the hindlimb. RIC was either delivered once 18h before, once 4hr acutely after or daily for 28 days beginning day 5 after endothelin-1 (ET-1) stroke. Infarct volumes were assessed 24hrs after stroke using MRI. To determine if RIC efficacy varied across stroke size, a hierarchical cluster analysis was used to divide rats into subgroups based on stroke size (small/large). RIC was effective in ET-1 which produced smaller strokes (“small”:5.2%, “large”:18.0% of hemisphere) than MCAO (~34%). This is more comparable to injury sizes seen clinically (4.5-14.0%). “Small” (42±4mm3) strokes were reduced by 39% (p=0.010, d=0.29) and “large” (146±8mm3) strokes were reduced by and 35% (p<.00001, d=1.41). Pre-RIC reduced infarct volume by 41% (p=<0.0001, d=0.92) versus 29% (p=0.009, d=0.43) in post-RIC. Interestingly, RIC is more effective in males, with double the infarct volume reduction of 46% (p<0.0001, d=0.94) compared with 23% (p=0.013, d=0.42) in females. Although RIC did not show restorative potential to improve motor stroke recovery, RIC is neuroprotective now with stronger clinically relevant evidence. RIC is effective across stroke models, stroke sizes and sex. Application of RIpreC to prevent stroke following a transient ischemic attack or recurrent stroke (especially in males with “large" strokes) would have the greatest potential.

Covert stroke (CS) comprises lesions in the brain often associated by risk factors such as a diet high in fat, salt, cholesterol and sugar (HFSCS). Developing a rodent model for CS incorporating these characteristics is useful for developing and testing interventions. The purpose of this thesis was to determine if magnetic resonance (MR) can detect brain abnormalities to confirm this model will have the desired anatomical effects. Ex vivo MR showed brain abnormalities for rats with the induced lesions and fed the HFSCS diet. Spectra acquired on the fixed livers had an average percent area under the fat peak relative to the water peak of (20±4)% for HFSCS and (2±2)% for control. In vivo MR images had significant differences between surgeries to induce the lesions (p=0.04). These results show that applying MR identified abnormalities in the rat model and therefore is important in the development of this CS rodent model.

Covert stroke (CS) comprises lesions in the brain often associated by risk factors such as a diet high in fat, salt, cholesterol and sugar (HFSCS). Developing a rodent model for CS incorporating these characteristics is useful for developing and testing interventions. The purpose of this thesis was to determine if magnetic resonance (MR) can detect brain abnormalities to confirm this model will have the desired anatomical effects. Ex vivo MR showed brain abnormalities for rats with the induced lesions and fed the HFSCS diet. Spectra acquired on the fixed livers had an average percent area under the fat peak relative to the water peak of (20±4)% for HFSCS and (2±2)% for control. In vivo MR images had significant differences between surgeries to induce the lesions (p=0.04). These results show that applying MR identified abnormalities in the rat model and therefore is important in the development of this CS rodent model.

Indiana University-Purdue University Indianapolis (IUPUI) / The vascular network highly coordinates with the central nervous system (CNS) on exchanging oxygen, nutrients and information transfer. The resemblance of the two systems at anatomical, cellular, and molecular levels also demonstrates their interdependence. The spinal cord is an integrated part of the CNS. Traumatic spinal cord injury (SCI) causes rapid systemic vascular responses and local neural tissue damage at the initial phase. The early disruption of the spinal vasculature breaks the supply-and-demand balance and facilitates the deterioration of the spinal cord tissue and functional deficits. Therefore, it is important to dissect the mechanism underlying vascular injury-mediated histological and functional consequences in order to develop potential therapeutic strategies. To visualize dynamic vascular changes after an acute SCI, a novel duo-color in vivo imaging technique was successfully developed in adult rats at the cervical level. This technique overcomes previous technical hurdles allowing real-time observation of vascular changes in live animals. Correlated with histological measures, in vivo vascular outcomes revealed a temporospatial relationship with neuronal and axonal loss, myelin disruption, inflammation, and glial responses. For the first time, we defined a “transitional zone” where significant blood vessel dilation and vascular leakage were observed simultaneously with vascular changes occurred at the injury epicenter acutely after SCI. These vascular changes at the transitional zone happened before any other cellular damage after SCI, suggesting a time window to prevent further neuronal damage in this region. Targeting the observed vascular leakage can work as a proof of concept that early vascular dysfunction contributes to the secondary neural tissue damage. Indeed, intravenous delivery of ferulic acid conjugated with glycol chitosan (FA-GC) to the injured sites immediate after SCI resulted in reduced vascular leakage, ventral horn neuronal loss, and partial recovery of forelimb function following a clinically-relevant contusive SCI at the 7th cervical spinal cord level. In conclusion, this work elucidated a novel role and mechanism of early vascular damage in the “transitional zone” prior to the secondary damage of neural tissue in this region and provided a novel treatment strategy for early neuroprotection and functional recovery. / 2021-11-04

Evidence suggests that there is an immunological response of the recipient to xenograft bioprosthetic heart valves. Information on the impact of HMG-CoA reductase inhibitors (statins) and their anti-inflammatory properties on bioprosthetic valve failure remains limited. We sought to examine the efficacy of statin therapy in a rodent model of bioprosthetic valve implantation. To mimic the human scenario, fresh or glutaraldehyde-fixed aortic valve root conduits from Lewis rats or Hartley guinea pigs were microsurgically implanted intravascularly into the infra-renal aorta of Lewis rats. The syngeneic control group consisted of a fresh rat valve conduit implanted into a rat. The xenogeneic control group consisted of a glutaraldehyde-fixed guinea pig valve conduit implanted into a rat. Treatment groups consisted of xenogeneic groups treated with either daily steroids or statins. Overall, steroid treatment attenuated the inflammatory response observed within the xenogeneic glutaraldehyde-fixed valve conduits. Treatment with statins did not decrease this inflammatory response.

Evidence suggests that there is an immunological response of the recipient to xenograft bioprosthetic heart valves. Information on the impact of HMG-CoA reductase inhibitors (statins) and their anti-inflammatory properties on bioprosthetic valve failure remains limited. We sought to examine the efficacy of statin therapy in a rodent model of bioprosthetic valve implantation. To mimic the human scenario, fresh or glutaraldehyde-fixed aortic valve root conduits from Lewis rats or Hartley guinea pigs were microsurgically implanted intravascularly into the infra-renal aorta of Lewis rats. The syngeneic control group consisted of a fresh rat valve conduit implanted into a rat. The xenogeneic control group consisted of a glutaraldehyde-fixed guinea pig valve conduit implanted into a rat. Treatment groups consisted of xenogeneic groups treated with either daily steroids or statins. Overall, steroid treatment attenuated the inflammatory response observed within the xenogeneic glutaraldehyde-fixed valve conduits. Treatment with statins did not decrease this inflammatory response.

Les tauopathies sont des maladies neurodégénératives caractérisées par l’agrégation de protéine Tau. Mis à part ce marqueur pathologique commun, les tauo-pathies présentent une grande diversité, tant sur le plan clinique qu’anatomo-pathologique, qui pourrait résulter de mécanismes pathologiques différents. Un mécanisme commun pourrait impliquer de petits agrégats de type oligomériques comme étant l’espèce toxique dans les tauopathies.L’objectif de ce projet est de déveloper plusieurs modèles de tauopathies, génétiques ou sporadiques, par transfer de gène de la protéine Tau humaine à l’aide de vecteurs adéno-associés. Trois vecteurs sont utilisés, chacun induisant un phénotype spécifique. Tout d’abord, la surexpression de hTAUWT induit une hyperphosphorylation de la protéine associée à une forte mort neuronale, mais en absence d’agrégation.Sa co-expression avec le peptide pro-agrégant TauRD-ΔK280 dans le groupe hTAUProAggr promeut son agrégation, ce qui est associé à des effets neuroprotecteurs. La construction hTAUP301L induit à la fois la formation d’agrégats solubles et matures, associée à un niveau intermédiaire de toxicité. Ces résultats supportent l’idée que les formes solubles oligomériques jouent un rôle crucial dans la pathologie associée à Tau.Ces modèles rapides, obtenus après expression de Tau à des niveaux similaires, récapitulent donc la variabilité phénotypique observée dans les tauopathies humaines. Ils devraient servir dans le futur pour étudier les mécanismes sous-tendant la toxicité des différentes espèces de Tau. Ils pourraient aussi être utiles pour étudier la spécificité et la sélectivité de composés développés pour l’imagerie de Tau par tomographie par émission de positon. / Tauopathies are neurodegenerative diseases characterized by the aggregation of Tau protein. Despite this common hallmark, tauopathies exhibit a wide variety of clinical and anatomo-pathological presentations, which may possibly result from different pathological mechanisms. One hypothesized common mechanism, however, implicates small oligomeric aggregates as drivers of Tau-induced toxicity.The aim of this project was to develop models of sporadic and genetic tauopathies, using adeno-associated viruses to mediate gene transfer of human Tau to the rat brain. Three different constructs were used, each giving rise to a specific phenotype. First, hTAUWT overexpression led to a strong hyperphosphorylation of the protein which was associated with neurotoxicity in absence of any significant aggregation. Its co-expression with the pro-aggregation peptide TauRD-ΔK280 in the hTAUProAggr group strongly promoted its aggregation, with neuroprotective effects. hTAUP301L construct led to aggregation into soluble species as well as mature aggregates accompanied with an intermediate toxicity. Those results support the hypothesis that soluble oligomeric species are key players of Tau-induced neurodegeneration.Those fast developing models, obtained through similar overexpression of human Tau, thus recapitulated the phenotypic variability observed in human tauopathies. Those should prove useful in the future to study mechanisms underlying the toxicity of various Tau species. Those could also serve to study the specificity and selectivity of Tau-directed tracers for positon emission tomography (PET) imaging.

Endogenous neural stem/progenitor cells therapy is one of the most advanced clinical trial worldwide. Generally, drug is given to the targeted area through the traditional strategies such as intraventricular or intravenous delivery method. However, those methods always come with undesired side-effects such as over-dose of drug and offensive injection are not applicable to the large-scale clinical application. In this study, the clinical feasibility of blended biosynthesized cellulose duraplasty was studied. Our results showed that physical properties of BBC can be controlled through the optimized fabrication process. In addition, the time length of Middle cerebral artery occlusion rat model was tested through the 60 vs 90 mins occlusion time behavioral assessments of rat and the data indicated that 60 mins length can induce significant motor functional impairment. Finally, the EGF & EPO-loaded BBC duraplasty was implanted over the removed area and the ELISA test revealed that BBC duraplasty can release and delivery the growth factors to the targeted area (subvertical zone) at least 3 days after implantation. In summary, our BBC duraplasty is showing the potential prospection to be a clinical-applicable duraplasty to replace the traditional commercial duraplasty in the future stroke recovery therapy.

Alcohol is the most abused drug in the United States and most frequently performed assay in forensic laboratories. Alcohol is routinely present in biological specimens from fatal residential fires and forensic toxicologists must interpret if these individuals are impaired by determination of their blood alcohol concentrations on post-incineration blood collected at autopsy. There is no known data available to confirm or refute blood alcohol concentrations and impairment in fire-related deaths. Ethyl glucuronide (EtG), a non-volatile minor ethanol metabolite, may provide a better biomarker for ethanol consumption prior to burn injury. The literature does not address the possibility that ethanol or EtG concentrations are altered in fire deaths. A Sprague Dawley rat model was employed to determine if ethanol and EtG concentrations in blood, liver, heart, and kidney were altered after burn injuries using two incineration models with varying durations and temperatures. Blood and tissues were analyzed for ethanol by gas chromatography and EtG by enzyme immunoassay. Other measurements including organ weights, lower hindquarter weights, and blood glucose concentrations were chosen for analysis to determine the mechanism by which the blood and organ ethanol and EtG concentrations are altered in burnt corpses. The rodent provided an excellent model for studying the biotransformation of ethanol to EtG and the effects of burn injury on ethanol and EtG concentrations. Our study revealed that blood ethanol concentrations were not significantly altered by burn injury but tissue ethanol concentrations were altered by burn injury. EtG concentrations were found to be altered in blood and tissue specimens in both incineration models. Our data suggest that the change in ethanol and EtG concentrations may be correlated to higher core body temperatures from burn injury and not changes in organ weight. Determining if blood ethanol concentrations are altered in burnt corpses is important for forensic toxicologists to conclude if victims were impaired at the time of death. The knowledge gained from these experiments will help forensic toxicologists by confirming the current interpretation that blood ethanol concentrations are not altered in fire deaths and provide a better understanding for the interpretation of impairment in burn deaths.