E. coli bacteriophages Lambda and PF-2 display differing patterns of adsorption rates following treatment with hydrogen peroxide.
Madeleine Bruderer and Archibald Landrum
The intestinal immune system is tasked with maintaining a balance between tolerating the intestinal microbiota and defending against invading pathogens. The intestinal microbiota is a complex community comprised of bacteria, fungi, archaea, and viruses, namely bacteriophage. During times of intestinal inflammation, such as in IBD, bacterial communities are seen to be greatly impacted. However, much less is known about how this intestinal inflammation impacts bacteriophage communities, and more specifically how innate inflammatory products influence their infection potential. The goal of this study was to evaluate the impact of innate immune products, specifically hydrogen peroxide, on the adsorption potential of a temperate and virulent bacteriophage both specific to E. coli. To do this, adsorption kinetics of a virulent and temperate bacteriophage were measured in the presence of varying doses of hydrogen peroxide. For PF-2, a virulent bacteriophage, hydrogen peroxide caused a delay in adsorption in a dose-dependent manner. However, for Lambda, a temperate bacteriophage, no effect on adsorption was observed regardless of dose.
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New Petrologic and Single Crystal 40Ar/39Ar Data for the Western Blue Ridge and Implications for Understanding the Extent and Conditions of Peak Alleghanian Metamorphism
H. Cole Burton
The Western Blue Ridge (WBR) province has a polymetamorphic history representing Ordovician, Devonian and Carboniferous stages of Appalachian orogeny. Regional U/Pb ages for zircon and monazite commonly record Middle to Late Ordovician ages; 40Ar/39Ar ages for amphiboles and micas are generally Silurian to Carboniferous, and have variously been interpreted as recording cooling following Ordovician metamorphism or superimposed stages of Acadian and/or Alleghanian metamorphism. The Murphy Synform contains the structurally and stratigraphically highest metamorphic sequences of the WBR, and it has been correlated with the Talladega Belt of Georgia and Alabama, that contains fossils as young as Early Mississippian. We find that an amphibolite from the Marble Hill Hornblende Schist of the Murphy Group (collected from cored drilled near Tate, GA) contains pargasitic hornblende with the formula (Na.35, K.31)Ca1.89(Fe2.44,Mg1.97,Al0.64)(Al1.53Si6.47)Si8O22(OH)2 and a slight chemical zoning from core to rim reflecting increasing Tschermak substitution.40Ar/39Ar incremental heating analysis of single pargasitic hornblende crystals yields a spectrum that commences with ages of ~ 305 Ma to define a plateau age of 318±2 Ma (7 steps with 80% of the 39ArK released, 2σ). The new data support regional structural, stratigraphic and paleontological data indicating rocks of the Murphy Synform — and the WBR from the Talladega Belt to Great Smoky Mountain National Park — experienced Alleghanian folding, peak metamorphism and isograd development. The content of this abstract for presentation within Auburn University is reproduced in its entirety from an original submission by Burton and Hames (2020) to the Annual Meeting of the Geological Society of America.
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Electromyographic Analysis Of Shoulder Rotation Strength Testing Positions
Molly M. Cassidy
Shoulder strength is essential for upper extremity function during overhand sport tasks. Clinicians perform shoulder rotational strength tests in a variety of positions to examine shoulder function; however, shoulder function may vary across positions. PURPOSE: The purpose of this study was to compare measures of shoulder function (peak torque and muscle activation) between two shoulder rotational strength testing positions. METHODS: Nine physically active participants (6 females, 3 males, age: 21.4±2.4 y, height: 170.4±7.9 cm, weight: 71.6±10.1 kg) performed isometric shoulder internal and external rotation strength tests using an isokinetic dynamometer in the following positions: (1) supine with arm abducted at 90° in the frontal plane and (2) seated with arm abducted at 90° in the frontal plane and internally rotated 45°. The elbow was flexed 90° in both positions. Electromyographic data were collected on the posterior (PD) and anterior deltoid (AD). Maximum voluntary contraction (MVC) testing established baseline muscle activity to which subsequent trials were normalized. A 2(position) x 2(direction) repeated measures analysis of variance (RM·ANOVA) compared torque values between testing positions for external and internal rotation tests. A second 2(muscle) x 2(position) RM·ANOVA compared muscle activation (%MVC) between testing positions for AD and PD muscles. RESULTS: The first RM∙ANOVA did not reveal a significant position by direction interaction. The second RM∙ANOVA did reveal a significant muscle by position interaction [F(1, 8) = 8.700, p = 0.018]. Post hoc analysis showed a difference between supine (mean: 20.4, SD: ± 2.6) and seated (mean: 32.6, SD: ± 2.4) positions for AD activation, where greater activation was measured in the seated position (p = 0.011). CONCLUSION: Although peak torque differences were not observed between positions, clinicians should note the AD may have greater contribution to overall shoulder strength in the seated compared to the supine position.
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Rapid Diagnosis of Infectious Diseases and Environmental Compounds Detection
Mathew Dudich
IM-MS (ion mobility - mass spectrometry) can be utilized to analyze complex mixtures rapidly and efficiently. Doing so can identify exactly what is in a certain sample. Herein, we have used IM-MS platform coupled with Liquid Chromatography (LC) separation to analyze a mixture of sulfonamides. This study was conducted to demonstrate the rapid separation power of IM-MS method using 4 sulfa drugs compared to LC-MS analytical method which is time consuming. Time-efficiency was the goal. Sulfonamides - or more commonly "sulfa drugs" - are antibiotics most notably used for treatment of bacterial infections including eye infections and urinary tract infections. These drugs work by attacking a mechanism that creates folic acid in bacteria and create a bacteriostatic effect. The 4 sulfa drugs under this study are sulfadimethoxine, sulfachloropyridazine, sulfamethazine, and sulfamethizole. An LC-MS analytical gradient method was developed to separate these 4 sulfa drugs. The starting gradient employed span from 10% to 90% acetonitrile within 10 minutes to elute all the sample components. The resolution and separation time were optimized by varying the solvent strength, flow rates, and column temperature, which in turn resulted in an analysis time of 3 minutes. Using the IM-MS method however, all 4 sulfa drugs were separated within only one minute. Creation of a faster method contributes to faster analysis of complex sample mixtures. Research of this kind is paramount for analyzing the makeup of drugs quickly in order for them to reach consumers in a timely fashion.
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Novel Streptomyces species from marine sponges produce metabolites that kill methicillin-resistant Staphylococcus aureus (MRSA)
Dory Fawwal
Streptomyces species are well-known microbial producers of medically relevant antimicrobial compounds that are important for their survival in natural environments. In an effort to characterize the antimicrobial potential of novel Streptomyces species S. poriferae and S. tardus derived from marine sediments and sponges, three Streptomyces isolates grown in various fermentation media were screened for activity against bacterial and fungal pathogens. Sequence-based bioinformatic analyses were also performed to compare the phylogenetic relatedness of the isolates to previously described Streptomyces species and to further explore the biosynthetic potential of the novel isolates by mining genomes for biosynthetic gene clusters (BGCs) that can encode natural products like antibiotics. Antimicrobial activity assays demonstrated that supernatants from the Streptomyces isolates inhibited the growth of Gram-positive pathogens including MRSA and in some cases inhibition was also observed for fungal pathogens. The antimicrobial activity observed was dependent on the duration of incubation and the fermentation media used. Furthermore, genome analyses revealed that the isolates harbored on average 30 BGCs, many of which were predicted to be unique, and phylogenetic analyses confirmed the novelty of the Streptomyces species. These results demonstrate that novel species from the genus Streptomyces were capable of producing antimicrobial metabolites active against human pathogens. These newly discovered antibiotics may be useful in treating human disease and work towards determining the structure of these antibiotics is ongoing.
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Identifying Heme Intermediates of Protein-based Cofactor Formation in
Catalase-Peroxidase (KatG)
Madeleine Forbes
Catalase-peroxidase (KatG) is a multifunctional, heme-dependent enzyme that catalyzes a unique combination of catalase and peroxidase activities. The enzyme aids in the detoxification of peroxides and serves as a key virulence factor for a range of bacterial and fungal pathogens. Cooperation of both activities cannot be achieved without two novel active-site structures: 1) a novel covalent structure formed between a Met, a Tyr, and a Trp (i.e., the MYW cofactor) and 2) an Arg residue near the active site whose conformation is pH-dependent (i.e., the Arg switch). However, both the mechanism of the MYW cofactor formation and the specific role of the Arg switch in that process are unknown. Pretreating the reconstituted enzyme with various equivalents of peracetic acid (prR418N) allowed for the formation of the cofactor to be monitored. Kinetics of the heme intermediates and of the reaction was measured by stopped-flow spectroscopy. Comparison of kinetic data and heme intermediates among the three different forms, rR418N, prR418N, and hR418N, showed a ferryl-oxo intermediate present in rR418N and absent in hR418N. Ongoing studies will compare this variant to the wild-type enzyme to further investigate the participation of the Arg switch in the formation of the MYW adduct.
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Troriluzole attenuates learning and memory deficits in 3xTG-AD mice
Emma Granberry
Alzheimer's disease (AD) poses a public health crisis, creating an urgent need for effective treatments and disease-modifying therapies. We tested a novel 3rd generation tripeptide prodrug of the glutamate modulator, riluzole, that offers improved bioavailability, safety, and dosing. We examined the effects of this drug, troriluzole, on synaptic function using the triple transgenic (3xTg) AD model. 3xTg mice received troriluzole (25 mg/kg/day) from 5-8 months of age. At eight months of age, drug-treated 3xTg mice were compared to vehicle-treated 3xTg and nontransgenic controls for alterations in synaptic plasticity using acute hippocampal slices. Basal synaptic transmission was reduced by 50-60% in vehicle-treated 3xTg slices compared to controls, and treatment partially attenuated these basal synaptic transmission deficits. To determine if these differences were related to presynaptic modifications, the probability of neurotransmitter release was examined by measuring paired pulse facilitation (PPF). PPF was significantly reduced in vehicle-treated 3xTg slices, indicating an increase in presynaptic release probability. Drug treatment restored PPF in 3xTg mice to that of controls. The amount of readily releasable pool (RRP) of synaptic vesicles was also significantly reduced in 3xTg mice, indicating increased vesicle docking. This treatment significantly increased RRP in 3xTg mice. We next measured long-term potentiation (LTP), a cellular correlate of learning and memory. LTP was reduced by 25% in vehicle-treated 3xTg mice compared to controls. Notably, drug treatment completely restored the synaptic plasticity deficits observed in 3xTg mice. These results highlight the potential of troriluzole as a novel therapy for AD.
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The role of FMO3 in Nonalcoholic hepatic steatosis
Victoria Jiminez
Type 2 diabetic patients display abnormally high levels of metabolic products associated with gut dysbiosis. One such metabolite, Trimethylamine N-Oxide (TMAO), is related to increased incidence of cardiovascular diseases (CVD) in human patients. Trimethylamine (TMA), a gut metabolite, comes from the metabolic degenerative product that is produced from gut microbial metabolism. Oxidation of TMA to TMAO is observed in elevated levels of diabetics and obese patients and has a direct correlation with increased risk for major adverse cardiovascular events. Mechanistically, the liver enzyme flavine-containing monooxygenase 3 is significantly involved in development of TMAO and hepatic gluconeogenesis. We predict that FMO3 is significantly involved in hepatic steatosis and hepatic insulin resistance. We have observed in leptin deficient (db/db) diabetic mice an increase (>3fold) of FMO3 when compared to wild type age (4 months) matched mice. Further in human liver cell lines (HEPG2) cells over expressing FMO3 (3 fold) resulted in significant lipid accumulation and reduced glucose uptake. These observations were compared to HEPG2 cells over expressing FMO1 and FMO5 which showed relatively no change in steatosis or changes in glucose uptake when compared to vehicle treated HEPG2 cells. Further we observed a significant increase in TMAO in FMO3 over expressing cells after treating with TMA. Lastly we observed an increase in steatosis in HEPG2 cells treated with TMAO. Our data is preliminary and further evaluation will determine the significance of FMO3 and TMAO on development of NAFLD. We are currently developing novel FMO3 inhibitors for potentially mitigating development of NASH.
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Associations between a baseball pitcher's arm slot and
shoulder joint loads
Luke Maddox
The shoulder is one of the most commonly injured areas among pitchers. A pitcher's arm slot has been shown to influence the loads placed on the pitching arm. Examining the relationship between a pitcher's arm slot and shoulder joint loads could provide additional insight into whether specific arm slots place pitchers at a greater risk of injury. Therefore, the purpose of this study was to investigate the associations between arm slot and peak shoulder joint loads during the baseball pitch. The kinematics of 22 youth baseball pitchers (75.0±7.9kg; 1.8±0.05m; 16.2±0.8yrs) were tracked using an electromagnetic motion capture system. Arm and trunk angles were examined at three different pitching events [foot contact (FC), maximum shoulder external rotation (MER), and ball release (BR)]. Shoulder joint loads were quantified using peak shoulder rotation torque and peak shoulder anterior force. Spearman's rank-order correlations were used to examine the associations between arm slot, trunk angle, and shoulder joint loads. Correlation analysis revealed no associations between shoulder joint loads and arm angle or trunk angle at FC, MER, or BR (all p > .212). The lack of association between shoulder joint loads and arm slot measures was surprising and in contradiction with previous research reporting that sidearm pitchers experience less shoulder anterior force than three-quarters and overhand pitchers. These findings suggest no specific arm slot consistently increases the loads placed on the shoulder. Pitchers should choose the arm slot that allows them to perform their best.
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Associations Between Cannabis Use Motives And Prodromal Symptoms
Regan Moss
Use of psychoactive substances is prevalent in people with prodromal syndromes and psychosis; in particular, cannabis use is rampant among schizophrenia patients. Importantly, however, cannabis may exacerbate psychotic symptom severity, resulting in worse outcomes among users. Although the co-occurence of cannabis use and psychosis is well-established in existing scholarship, few studies have highlighted the underlying motivations of cannabis use and their implications for subsequent psychopathology. In this study, we aimed to (1) assess cannabis use motivations among college students who completed the Marijuana Motives Measure (MMM), a five-factor explanatory model of cannabis use motivations (i.e., conformity, coping, enhancement, expansion, and social use motives) and (2) explore associations between cannabis use motives and scores on the Prodromal Questionnaire-Brief (PQB) Version, which measures prodromal symptoms. Confirmatory Factor Analysis (CFA) was used to assess the fit of the MMM five-factor model. In addition, simple linear models were used to explore associations between cannabis use motives and prodromal symptoms. In exploratory assessments, we highlighted the associations between MMM factors and cannabis use characteristics (onset age, use severity) as indicated by the Marijuana Smoking History Questionnaire (MSHQ). Regression analyses highlighted the five factors as predictors between cannabis motives and prodromal symptoms. In particular, Expansion (i.e., perceptual and cognitive enhancement) was positively associated with prodromal symptoms.. Determining relationships between cannabis use and prodromal symptom severity is an important step toward understanding the nuanced relationship between cannabis and psychosis.
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Immunologic and stress responses of hatchling American alligators (Alligator mississippiensis) exposed to ecologically relevant estrogen levels
Regan Moss
Elevated estrogen concentrations are commonly observed in many ecological systems. Given the role of estrogen in mediating stress (the process by which environmental demands strain an organism's adaptive capacity), it is critical to understand how acute exposure to environmental estrogens affect the stress and immune responses of wildlife. We experimentally examined the impact of environmentally relevant concentrations of estrogen on circulating glucocorticoid levels and several innate immune parameters in Alligator mississippiensis. This long-lived top-tier predator provides a model system for assessing the physiological effects of estrogenic compounds. Hatchling alligators were randomly placed in one of two treatment groups where different concentrations of estrogen (E2) were delivered via their diet; low E2 (0.5 µg/kg E2 , n = 8) or high E2 (1 µg/kg E2 group, n = 8), and compared to a control group (no E2, n = 7). Several biomarkers of elevated stress were used to monitor the impact of E2: white blood cell (WBC) counts, glucocorticoid levels, and general antibody response. Natural antibody levels were greater in individuals from the high and low E2 treatments compared to the control individuals. Mean H:L (heterophils: lymphocytes) ratios and blood estradiol concentration were significantly higher in the experimental groups than those of control animals. These results suggest that individuals exposed to environmental estrogen concentrations may exhibit increased immune and stress responses. Future studies may expand on this by monitoring biomarkers in wild populations.
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Novel PPAR-delta-gamma agonist mitigate hepatic steatosis via hepatocyte growth
factor signaling.
Christopher Purvis
Introduction: The clinical correlation between the high incidence of diabetes type 2 and Nonalcoholic fatty liver disease (NAFLD), is emerging as the most significant form of chronic liver disease in the world today. Therefore, there exists a critical need to develop novel therapies to help mitigate the development of NAFLD and its progression towards NASH.
Hypothesis: We predict that AU580, a novel dual PPAR-delta-gamma agonist developed in the Amin lab, mitigates development of hepatic steatosis in a rodent model of diabetes (db/db – leptin receptor deficient).
Methods: Db/db mice were given compound AU580 I.P. for 30 days (5mg/kg). Livers were extracted and histologically scored. Hepatic steatosis was developed using BSA-conjugated fatty acid in HEPG2 human liver cell line. Mouse liver and cell culture models of hepatic steatosis was measured by Oil-redO. HGF and downstream signaling was assessed by western analysis.
Magnetic resonance Imaging was accomplished using the 7T to measure levels of steatosis.
Results: We observed by western analysis enhanced hepatocyte growth factor (HGF) and p-cMET signaling in livers from AU580 treated db/db mice and human liver cell culture lines (HEPG2 cell). This signaling helps explain the mechanism for reduced lipid accumulation in our mice as measured by MRI, triglyceride measurements and Oil redO staining.
Conclusion: Our data is preliminary and further evaluation will determine the significance of AU580 on development of NAFLD and potentially for mitigating development of NASH. In this line of investigation, we are currently exploring the role of AU580 mitigating inflammation and fibrosis.
Hypothesis: We predict that AU580, a novel dual PPAR-delta-gamma agonist developed in the Amin lab, mitigates development of hepatic steatosis in a rodent model of diabetes (db/db – leptin receptor deficient).
Methods: Db/db mice were given compound AU580 I.P. for 30 days (5mg/kg). Livers were extracted and histologically scored. Hepatic steatosis was developed using BSA-conjugated fatty acid in HEPG2 human liver cell line. Mouse liver and cell culture models of hepatic steatosis was measured by Oil-redO. HGF and downstream signaling was assessed by western analysis.
Magnetic resonance Imaging was accomplished using the 7T to measure levels of steatosis.
Results: We observed by western analysis enhanced hepatocyte growth factor (HGF) and p-cMET signaling in livers from AU580 treated db/db mice and human liver cell culture lines (HEPG2 cell). This signaling helps explain the mechanism for reduced lipid accumulation in our mice as measured by MRI, triglyceride measurements and Oil redO staining.
Conclusion: Our data is preliminary and further evaluation will determine the significance of AU580 on development of NAFLD and potentially for mitigating development of NASH. In this line of investigation, we are currently exploring the role of AU580 mitigating inflammation and fibrosis.
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Evaluating the Effects of Chemotherapeutics on Dopaminergic Cells
Anna Solomonik
Chemotherapy-induced cognitive impairment, also known as "chemo brain", is a medical complication of cancer treatment that is characterized by a general decline in cognition. It affects visual and verbal memory, attention, complex problem-solving skills, and motor function. It is estimated that one-third of patients who undergo chemotherapy treatment experience cognitive impairment. Alterations in the release and uptake of dopamine and serotonin neurotransmitters that play important roles in cognition could potentially contribute to impaired intellectual performance in those impacted by chemo brain. Chemotherapeutics can increase oxidative stress, decrease mitochondrial function, and increase apoptosis which causes dopaminergic neurodegeneration ultimately leading to motor disorders such as Parkinson's disease. This study was done to investigate in vitro dopaminergic neurotoxic effects of the chemotherapeutics Doxorubicin and Cyclophosphamide. Rat dopaminergic neuronal cells (N27) were used for in vitro studies to assess the neurotoxicity. Cyclophosphamide and Doxorubicin exhibited dose-dependent and time-dependent in vitro dopaminergic neurotoxicity. They induced oxidative stress and apoptosis (increased BAX expression) in the dopaminergic neurons without affecting the mitochondrial functions. With regards to oxidative stress, Cyclophosphamide and Doxorubicin increased the generation of ROS and nitrite content resulting in lipid peroxidation. They also depleted glutathione, decreased catalase activity, and increased glutathione peroxidase, SOD, and MAO activity. Future studies to assess the in vivo neurotoxic effects of Cyclophosphamide and Doxorubicin using a rodent animal model will be performed. Therefore, this study supports the need for additional neurochemical, behavioral, and biochemical analysis to identify the underlying mechanisms of chemotherapy-induced cognitive disorders.
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KSHV viral copy number
Jake Tatum
Kaposi sarcoma-associated herpesvirus (KSHV) is an oncogenic herpesvirus that displays two phases of infection cycle. During latency the viral DNA persist as an episome attached to the host chromatin and limited expression of viral genes. During the lytic phase, the full repertoire of genes is expressed, and viral progeny are produced. The goal of my project is to set up a quantitative real-time PCR assay aimed at measuring the KSHV episome load in naturally infected and recombinant cell lines at the latent and lytic stages of KSHV infection. My model system is KSHV positive body cavity-based lymphoma cells (BCBL-1), which allows the controlled chemical manipulation of KSHV latent and lytic stages of infection. I am using two reference plasmids to create standard curves for my assay. These include a plasmid that codes for viral lytic gene K6, which allows me to estimate the viral episome copy number, and another plasmid that carries the endogenous retrovirus gene ERV-3, which allows to estimate the number of cells in analyzed samples. The PCR values (CT) obtained for my experimental samples will be compared to the created K6 and ERV-3 standard curves to deduce the total viral copy number in each experiment. This assay will be used in the lab to address how specific mutations introduced within KSHV genome affect viral replication and progeny virion production.
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Specific soybean cultivar growth is improved by co-inoculation of seeds with
Bacillus velezensis
Dylan Warner
There is a need to improve sustainability in agriculture in reducing the indiscriminate use of fertilizers and antibiotics. While the use of biological agents like beneficial bacteria to improve plant growth and reduce disease has increased in recent years, these "probiotics for plants" often fail in field trials because the living microorganism does not survive when introduced into a complex microbiota associated with the plant rhizosphere. As a potential solution, our previous studies have shown that the plant growth promoting rhizobacteria (PGPR) within the species Bacillus velezensis can grow using pectin, a complex carbohydrate produced by plant roots. By coating soybean seeds with the spores of B. velezensis (the probiotic) and pectin-rich orange peel (OP) powder (the prebiotic), we observed significantly improved effects on soybean plants that had increased root and shoot growth. We refer to this treatment as a symbiotic because it combines both prebiotic and probiotic treatments. Surprisingly, from this symbiotic we also observed a large increase in the number of nodules in soybeans treated with both B. velezensis spores and OP. The previous research was conducted only in a single soybean cultivar. In the current study, we selected 20 commercial soybean cultivars in order to evaluate cultivar responsiveness to this seed treatment that included both B. velezensis AP193 and OP, whereas control seeds were uninoculated. Once they reached the R2-R4 growth phase they were harvested. These data suggest there is significant variation among soybean cultivars as to their response to the B. velezensis and OP symbiotic treatment.
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Changes in vascular collagen density and cerebral blood flow in cats with a phosphoprotein enriched in astrocytes 15 kDa (PEA15) loss-of function mutation.
Anna Paige Wilson
Introduction: Phosphoprotein enriched in astrocytes (PEA15) is an important intracellular signaling molecule that can affect brain development in gyrencephalic species. In domestic cats, PEA15 deficiency leads to cerebral dysgenesis that presents as motor and sensory abnormalities. Preliminary RNAseq data suggests that PEA15 deficient cats have increased gene transcripts associated with endothelial cells and collagen deposition in their brain which may contribute to altered vascular perfusion during development.
Hypothesis/Objective: We predict that PEA15 deficiency results in increased collagen surrounding cerebral vessels, which may alter cerebral blood flow and perfusion.
Methods: In order to evaluate vascular collagen deposition, we stained histological sections of the cerebral cortex with Masson Trichrome and collected images of sub-meningeal cerebral vessels. A morphometric analysis software, QUPATH, was used to analyze vessels from four unaffected (PEA15+/+) and six affected (PEA15-/-) cats. A total of 3-5 vessels were measured in each brain tissue section. A standard g-ratio approach was used to evaluate the amount of collagen to the total vessel area. This ratio normalizes the region of interest (collagen) to the total area of the vessel to account for differences in vessel size.
Conclusion: There was a significant increase in the collagen stained regions of sub-meningeal cerebral vessels in affected cats compared to unaffected (p = 0.0381). This increase could lead to decreased cerebral perfusion during development and contribute to the cerebral dysgenesis observed in PEA15 deficient cats. Future studies will compare collagen thickness to in vivo brain perfusion.
Hypothesis/Objective: We predict that PEA15 deficiency results in increased collagen surrounding cerebral vessels, which may alter cerebral blood flow and perfusion.
Methods: In order to evaluate vascular collagen deposition, we stained histological sections of the cerebral cortex with Masson Trichrome and collected images of sub-meningeal cerebral vessels. A morphometric analysis software, QUPATH, was used to analyze vessels from four unaffected (PEA15+/+) and six affected (PEA15-/-) cats. A total of 3-5 vessels were measured in each brain tissue section. A standard g-ratio approach was used to evaluate the amount of collagen to the total vessel area. This ratio normalizes the region of interest (collagen) to the total area of the vessel to account for differences in vessel size.
Conclusion: There was a significant increase in the collagen stained regions of sub-meningeal cerebral vessels in affected cats compared to unaffected (p = 0.0381). This increase could lead to decreased cerebral perfusion during development and contribute to the cerebral dysgenesis observed in PEA15 deficient cats. Future studies will compare collagen thickness to in vivo brain perfusion.
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