It is well documented that pet cats develop age-related diseases similar to humans with chronic age-related diseases, including Alzheimer’s disease (AD). Since pet cats live in the same environment as their owners and by extension are subjected to the same environmental stressors, older pet cats are an excellent mammalian model to study therapeutic targets to slow or reverse brain aging. However, aging within the brains of pet cats is not well characterized, partly because valid reagents have not been identified. This study was designed to test several human-specific antibody reagents that identify non-neuronal cells, aging pathways, and Aβ amyloid and phosphorylated tau (pTau) seen at autopsy in brains from patients with AD. Archived brain samples, collected from pet cats at autopsy, were graciously provided by the veterinary pathology departments at University of California Davis campus and University of Pennsylvania. Immunohistochemistry staining was done to detect: 1) Microglia, a non-neuronal inflammatory reactive cell type, using an IBA1-specific marker; 2) An inflammatory pathway using an MCP-specific marker; 3) Amyloid plaques using E610, an Aβ42-specific marker; and 4) pTau fibrillary tangles using AT8, a pTau-specific marker. A digital imaging software program was used to generate a heat map to visualize staining and quantify results. It was found that brain samples from older pet cats had increased inflammation as determined by high staining intensity of microglia and MCP1. Brains from several cats showed evidence of amyloid plaques and pTau tangles. These observations suggest that the human-based reagents tested can identify analogous cell types, pathways, and pathogenic components of AD in brains from pet cats. These prototype reagents can now be used to begin the task of characterizing neuropathology in deceased pet cats donated to the Cat Alzheimer’s disease Program at the University of Washington.

Early-Stage Alzheimer’s Disease (ESAD) is characterized by the development of beta-amyloid aggregates (Aβ42) and phosphorylated tau (pTau) leading to mild cognitive decline and variable personality changes. Because specific diagnostic criteria have not yet been established for ESAD at middle age, there is no way of knowing who might be susceptible and who might be resilient to more severe neuropathology and dementia in later years. The geroscience concept assumes pathways associated with aging are also associated with age-related diseases including ESAD. Therefore, a simple skin biopsy procedure shown to predict resilience to aging in middle-aged mice should be able to predict resilience to ESAD in middle-aged mice. An adeno-associated-viral (AAV) vector system carrying pathogenic components of AD, Aβ42, and pTau, was used to induce ESAD in 23-month-old C57BL/6 mice. Before receiving the AAV vector, 2 mm ear punch biopsies were performed, and the rate of closure was measured over 3 weeks. The study ended when mice were 26 months of age, and the closure rate for each mouse was calculated and correlated with behavioral and neuropathological features of EASD. Preliminary observations will help address the question of whether the healing rate of a simple skin wound can predict susceptibility to the burden of AAV-mediated ESAD. It is expected increases in physical resilience will be associated with increased wound closure, and thus, mice with increased wound closure will have greater resilience to the onset of ESAD neuropathology. This could have highly impactful implications for the early treatment of ESAD in human patients thus preventing the irreversible and fatal progression of dementia associated with late-stage AD. In addition, DNA from skin biopsy cores could be used to obtain DNA methylation signatures for determining biological age thus providing an enriched, translationally relevant data set.
 

Alzheimer’s Disease (AD) is a progressive brain disorder that debilitates memory, learning, and decision-making. Early-stage AD represents the initial phase where individuals are still able to function independently, but with increasing age, their condition steadily progresses to dementia and loss of independence. Because a significant number of the aging population is affected by AD, understanding the neuroinflammatory processes would help develop more effective strategies for treatment. Examining markers such as MCP-1 and TNF-alpha, known to be associated with inflammatory response, will help identify the modulatory processes that lead to mild cognitive impairment associated with early-stage AD. Subsequently, higher levels of inflammation markers within the brain leads to mild cognitive impairment. This research study involved 40 C57BL/6 mice, 20 males and 20 females (21 months old), retro-orbitally infected with 80 µL of neurotrophic AAV-AD vector or AAV-Sham for a duration of 2 months before humane euthanasia. Brains were collected, and specific regions were examined by immunohistochemistry (IHC) and digital imaging to assess the expression levels and distribution of the inflammation markers. Preliminary observations showed that hippocampal regions of the brain from mice with early-stage AD had higher staining intensity for MCP-1and TNF-alpha compared to respective areas in Sham mice, suggesting increased inflammation is a very early lesion that develops in the presence of AD pathogenic components that might be controlled by anti-inflammatory drugs. The preliminary data suggests that the characteristics of AD manifest in part due to the neuroinflammatory response of brain factors that change with onset AD.

Brain-derived neurotrophic factor (BDNF) plays a critical role in neuronal function with potential implications for cognitive health, including involvement in adult neurogenesis. A decline in BDNF levels is associated with mild impairments in learning and memory. The hippocampus, known for its involvement in learning and memory processes, serves as a focal point for investigation in the brain due to its responsiveness to environmental stimuli, including exercise. There is an existing knowledge gap concerning whether running promotes an increase in BDNF levels within the hippocampus at very old ages, despite BDNF's importance in neuronal function and its potential implications for cognitive health. This study was designed to investigate whether physical exercise influences BDNF levels in the hippocampus of aged mice. Aged C57BL/6 mice were allowed access to running wheels, or locked running wheels, for three days, after which their brains were collected, post-euthanasia for neuropathology assessment. Immunohistochemistry (IHC) was performed with an anti-BDNF antibody by measuring BDNF presence, since lack of BDNF levels signifies lost neurons. QuPath digital imaging techniques were employed to provide a quantitative measure of the potential impact of running on hippocampal BDNF expression. Both the average and the variance of total distance run during voluntary wheel running decreased with age. Elevated BDNF levels were observed in the hippocampus of running mice compared to sedentary counterparts. The study provides insight into the potential impact of exercise on neurotrophic support in the aging brain. Such findings suggest a beneficial effect of exercise on neurotrophic support in the aging brain, and indicates the need for further investigations into lifestyle stratergies for promoting resilience to brain aging and cognitive decline in older adults.

Alzheimer's disease (AD) is a neurodegenerative age-related disease characterized by the presence of amyloid-beta aggregates and hyperphosphorylated tau tangles. It has been well documented that cognitive decline and changes in age-related pathways are associated with disease progression. Mitochondria play an important role in degradation of amyloid protein through a mitochondrial protein-mediated quality control system. This pathway can break down with increasing age and lead to the overwhelming presence of amyloid, disrupting normal mitochondrial activity. This damage leads to the formation of more Aβ plaques and neuroinflammation, contributing to the pathogenesis of AD. Mitochondrial regulators may be potential therapeutic drug targets but models are needed to help identify and characterize them. In this regard, an Adeno-Associated-Viral (AAV) vector was used to induce AD protein expression in the brains of old mice. 40 Male and 40 Females mice aged 24 months were infected with either the AAV-AD or AAV-SHAM vector and given 3 months for expression of the proteins to build. Mice were euthanized and brain tissue collected into formalin, with the hippocampus cut into slides for immunohistochemistry (IHC). Data generated from these mice has shown trends in decreased synaptic integrity, increased inflammation and DNA damage associated with expression of the vector proteins. Utilizing the same model, this experiment aims to understand how expression of the AAV-AD proteins may be associated with known roles of mitochondria and characterized pathways in the early stages of AD. IHC was performed using antibodies specific for PITRM1, a mitochondria protein degradation regulator, and PINK1, responsible for mitochondrial-mediated cell death (mitophagy). Imaging software “ImageJ” will be used for quantitative analysis of the stains. This study will help clarify an association between varying levels of AD protein expression and mitochondrial regulation, providing valuable information for enhancing therapies aimed at preventing the progression of early stage AD.

Alzheimer's Disease (AD) is incredibly complex such that development of neuropathology and cognitive impairement is driven by multiple pathways. Therefore, targeting these pathways simultaneously, could provide a more effective treatment for AD compared to any single drug. Rapamycin, acarbose, and phenylbutyrate each have independent but overlapping effects on multiple pathways involved in cellular respones to pathogenic beta amyloid such as inflammation, glucose homeostasis, synaptic integrity, autophagy, and DNA damage. To test the safety and effectiveness of a cocktail of these three drugs, a proof of concept experiment was undertaken in transgenic mice carrying mutations for genes associated with early onset AD (5xFAD). These mice express neuronal amyloid plaques, a major feature of AD neuropathlogy. Transgenic and wild type mice were given either a control feed or feed containing the drug cocktail starting at 4 months of age and continued until 12 months of age. Medicated transgenic mice showed significantly less cognitive impairement in a spatial navigation learning task and reduced amyloid plaque levels in the hippocampal brain region compared to untreated transgenic mice. Immunohistochemistry will be used to identify specific biomarkers for inflammation, synaptic integrity, glucose homeostasis, autophagy, and DNA damage in the hippocampus of treated and untreated transgenic mice. Observation from this study will suggest the need to conduct additional preclincial experiments testing this specfic drug combination for a successful approach to treat Alzheimer's Disease. 

On the morning of September 10th, 1990, the body of seventeen-year-old Maria Soledad-Morales was found on the outskirts of San Fernando de Valle, the capital city of Catamarca province in Argentina. What followed in the aftermath of the murder and botched investigation was the Catamarcazo, one of the largest and most publicly salient protest movements in the decades following the Argentinian Dirty War, the military dictatorship that murdered 30,00 people between 1977 and 1983. In this project, I examine genealogies of resistance among women-led protest movements in Argentina from 1977 to 1992 and argue that linkages between movements contributed to a modern Argentinian understanding of gendered violence in the present day. Through analysis of newspapers, interview transcripts, and photographs, I link activist strategies of the 1977 - 1983 Mothers of the Plaza de Mayo with the Catamarcazo movement of 1990 - 1991. I discuss how the Catamarcazo drew and built upon the strategies of Las Madres and introduced consciousness of gendered violence to the Argentinian public in the first nationally acknowledged protest movement centered on violence against women. The purpose of this study is to establish and recognize the continual development of feminist activism within Argentinian history and shed light on the subversive, revolutionary tactics used to combat state repression and gendered violence. By analyzing histories of feminist activism in Argentina, we can gain a greater understanding of how the strategies of feminist movements are built upon and expanded over time, and how the strategies of a past movement can be modified to serve a current movement.

More than 53 million adults or 22% of the US suffer from chronic inflammation. Along with being a critical factor in the onset and progression of aging and cell senescence of the central nervous system, inflammation is also a central hallmark of neurodegenerative diseases and brain injury. Modulation of neuroinflammation has the therapeutic potential to decelerate aging processes in the brain. Studies consistently show that cytokines such as Interleukin (IL)-6, Tumor Necrosis factor (TNF)-α, Monocyte Chemoattractant Protein -1 (MCP-1), and their receptors, are upregulated in aged tissues and cells. This study investigated effects of GHK-Cu, a naturally occurring peptide that has regenerative properties, as a potential therapeutic measure to reduce the expression of these proteins. Mouse SIM-A9 microglial cells and N2A neuroblastoma cells were used as a model system, and bacterial endotoxin (Lipopolysaccharide, designated as LPS) was used as a stressor to trigger an inflammatory response. Cell morphology, viability, and cytokine ELISA assays including IL-6, MCP-1, and TNF-α, provided data suggesting that GHK-Cu peptide is a potent factor in enhancing resistance of neuronal and microglial cell lines to LPS-induced stress by reducing the expression of key inflammatory cytokines.Understanding the mechanisms by which GHK-Cu modulates inflammation can pave the way for the development of novel treatments targeting inflammation-associated diseases, aging mechanisms and various forms of dementia.

Mild cognitive decline with increasing age commonly affects millions of people beginning as early as middle age. It can progress to more severe levels of cognitive impairment including dementia associated with Alzheimer’s disease and irreversible brain damage with eventual death. Therefore, treatment before the onset of dementia would be the most effective way to prevent the devastating loss of normal daily living and death as an outcome. However, few drugs have been shown to be successful in preventing the progression of mild cognitive decline to more severe cognitive dysfunction. One candidate drug we are testing is the naturally occurring peptide GHK (glycyl-L-histidyl-L-lysine), which is known to have regenerative and anti-inflammatory properties in the brain. In order to test this peptide, we treated middle-aged male and female C57BL/6 mice with GHK as a copper complex (GHK-Cu) or saline using a novel intranasal atomizer daily for two months. We then conducted behavioral tests to assess learning and memory, and then mice were euthanized to collect brain samples for special stains for biomarkers of brain aging including the presence of non-neuronal microglia, brain-derived neurotrophic factor, and synapse integrity. Our preliminary observations from behavioral tests show that mice treated with intranasal GHK-Cu performed better in learning and memory tests than mice treated with intranasal saline. The brain aging biomarker tests I completed show that the neuropathology markers associated with aging are less severe in mice treated with intranasal GHK-Cu. Such a positive outcome provides the rationale to do further preclinical testing as a way to move toward clinical studies designed to treat mild cognitive decline and prevent the devastating progression of irreversible neurodegeneration.