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  Detection of Anorexia Nervosa in Primary Care: Effects of Patient Ethnicity, History of Mental Illness, Physician Gender, and Years of Experience.

  Ashley Higgins, Stacey C. Cahn, Robert A. DiTomasso, and Harry J. Morris

  Poster presentation accepted for the 47th Annual Association for the Advancement of Behavioral and Cognitive Therapies, Philadelphia, PA.

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  Introducing Blended Learning to Medical Students in a Clinical Training Environment

  Erik Langenau and Douglas Koch

  Third year medical students typically learn through supervised patient interaction in a clinical setting, often based in academic or hospital-based training sites. As one of the five medical schools in the Philadelphia area, PCOM strives to find the best training sties for its students in a number of specialties. Due to the increasing number of students in the region as well as a decreasing number of available training sites (as a result of reduced hospitalization rates and lengths of stay), finding suitable training sites for students has been a challenge. In order to better meet the academic needs of our students, new educational models are required. These models need to address a number of key challenges. 1.Inconsistent clinical exposure to patients whose demographics and presenting problems vary by site. 2.Inconsistent training and quality of clinical preceptors at each of the clinical sites. 3.Insufficient clinic training sites to accommodate expanding class size.

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  Analysis of autophagy and inflammasome regulation in neuronal cells and monocytes infected with Chlamydia pneumoniae: Implications for Alzheimer’s disease

  Brian J. Balin, Christine J. Hammond, Juliana Zoga, Ahmad B. Cader, Annette K. Slutter, Jonathan M. Anzman, Ian Kohler, Susan T. Hingley, and Denah M. Appelt

  Objectives: Our laboratory has been studying the role of infection with the obligate intracellular bacterium Chlamydia pneumoniae in sporadic late-onset Alzheimer disease (LOAD). This infection may be a trigger for the pathology observed in LOAD as a function of initiating changes in gene regulation following entry of the organism into the brain. As such, we are analyzing how this infection can promote changes in autophagy and inflammasome gene regulation as both have been shown to be altered in LOAD. Methods: Human SKNMC neuronal cells and THP1 monocytes were infected in vitro for 24-72 hrs with a laboratory strain of Chlamydia pneumoniae followed by RNA extraction, cDNA synthesis and analysis using Real-Time PCR microarrays for autophagy and inflammasome genes. Results: Gene expression for autophagy and inflammasome pathways was altered dramatically following infection. Genes encoding for co-regulation of autophagy, apoptosis, and the cell cycle that were significantly changed included: BCL2L1, FAS, PIK3CG, APP, and TP53. In addition, ATG3, and GABARAP, genes encoding for protein transport & ubiquitination and autophagic vacuole formation were significantly deregulated. Of the inflammasome genes, 4 NOD-like receptor genes were significantly up-regulated. IL-1beta, AIM2, CCL2, and CCL7 genes were all dramatically up-regulated in monocytes during the 72 hrs of infection. Conclusions: Our data suggest that Chlamydia pneumoniae-infected human SKNMC neuronal cells and THP1 monocytes exhibit specific changes in gene regulation for both autophagy and inflammasome pathways. These gene changes appear to correlate with pathologic changes previously reported in AD and further support the contention that infection with Chlamydia pneumoniae plays a role in LOAD pathogenesis.

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  What Factors Contribute to or Inhibit Publishing Among Medical Librarians

  Skye Bickett, Christine Willis, and Melissa Wright

  OBJECTIVE: To discover what factors influence librarians to publish or present their work. The study aims to determine what type of publications and presentations are most prevalent, what external factors impact the decision to publish or present, what internal factors affect the decision, and why one chooses to do one rather than the other or both.

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  Chlamydia pneumoniae infection of neuronal cells induces changes in calcium-associated gene expression consistent with Alzheimer’s disease

  Christopher Andrew Cappellini, Ahmad B. Cader, Keith G. Williams, Juliana Zoga, and Susan T. Hingley

  Background and Significance: Previous studies have shown that cells infected with Chlamydia pneumoniae (Cpn) exhibit altered gene expression consistent with that observed in Alzheimer’s disease (AD). Furthermore, AD neurodegeneration has been linked to dysregulation of intracellular calcium and calcium-related processes. Therefore, we hypothesize that one mechanism by which pathogenesis evolves in AD is through infection-induced changes in expression of calcium-related genes. Objectives: To determine if infection of neuronal cells with Cpn alters expression of calciumrelated genes associated with neurodegeneration. Methods: SK-N-MC neuronal cells were infected with Cpn (AR39 strain; MOI=1) for 3 to 72 hours, then calcium-related genes were screened with real-time PCR microarrays (SABiosciences PAHS-066). Results: Following infection, approximately 29 genes displayed regulation changes of 2-fold or greater, including genes pertaining to neurotransmitters, cell cycle and immune regulators, and other calcium-responsive elements. Genes involved in synaptic function and memory such as AREG, ATF3, EGR2 and GEM were initially up regulated, then fell to baseline or below by 72 hours. Many of the affected genes have been implicated in AD pathogenesis. Conclusions: Our data suggest that Cpn alters calcium-related gene expression in host neurons consistent with calcium dysfunction previously documented in AD. This study may elucidate how, in its effort to establish a favorable environment, Cpn could affect cellular processes that contribute to AD pathogenesis.

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  Pediatric Obesity Class: Teaching Kids How to Live Healthy Lifestyle

  Sara Wilson Reece

  Childhood obesity has reached epidemic proportions in the United States. At The Longstreet Clinic, PC, Diabetes Education and Medical Nutritional Therapy Department, and Pediatric Department have joined together to create the Pediatric Obesity Class to address the epidemic of pediatric obesity in the local community. In the class, the child and parent are taught about calories, how to develop healthy eating habits and become physically active as a family. Visual aids as well as hands on activities for both nutritional and physical activity components of class are utilized. Thus far, approximately 120 children along with their parent(s) have participated in the class. Pediatric department tracks post-class height and weight measurements. Currently, additional outcomes are being considered to measure the effectiveness of the class.

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  Implementation of Active Learning Components into an OTC Course

  Sara Wilson Reece and Nicole Phipps

  When faculty is preparing for didactic class, it is important for the faculty to consider that students only retain about 10% of what they hear, while they retain up to 90% of what they say or do.1 Active learning facilitates learning by enhancing a student’s learning experience in that he/she surpasses understanding of a subject into analysis, synthesis, and evaluation of the subject. Incorporation of active learning strategies such as minute writes and the use of i>clicker audience response system, have been shown to improve a student’s performance on free-recall quizzes and comprehensive exams up to two letter grades.

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  Autophagy and apoptotic genes implicated in Alzheimer’s disease are modulated following infection of neuronal cells with Chlamydia pneumoniae

  Denah M. Appelt, Ian Kohler, Annette K. Slutter, Juliana Zoga, Susan T. Hingley, and Brian J. Balin

  Background: The focus of the current studies was to determine the relationship between the molecular mechanisms interconnecting autophagy and apoptosis following Chlamydia pneumoniae infection in neuronal cells. Dysfunctions in apoptosis and autophagy have been implicated in the neurodegeneration associated with Alzheimer’s disease (AD). Autophagy in AD pathogenesis has been shown to play a role in amyloid processing through the endosomal-lysosomal system. Apoptosis may contribute to the neuronal cell loss observed in AD; however, there is limited evidence of the apoptotic process proceeding to terminal completion. Although Aβ1-42 has been shown to induce apoptosis in neurons and may be an early factor in AD, our previous investigations demonstrated that neurons infected with Chlamydia pneumoniae are resistant to apoptosis, and that Aβ1-42 is induced following this infection. Thus, these studies address infection as an initiator/trigger or inhibitor for the processes of autophagy and apoptosis observed in Alzheimer’s disease. Methods: SKNMC neuronal cells obtained from ATCC were infected with the AR39 strain of Chlamydia pneumoniae at an MOI=1 for 24, 48, and 72hrs and were analyzed using Real-time PCR arrays from SABiosciences specific for autophagy and apoptosis genetic markers. Results: Some major genes associated with apoptosis such as BID, DAPK1, TP53, TP73 were down regulated by 72hrs post-infection. Genes associated with the regulation of autophagic vacuole formation such as ATG3, ATG4B, ATG4C, ATG9A, ATG9B, ATG12, IRGM, and BECN1 were up-regulated within 72hrs post-infection. With regards to genes involved with co-regulation of autophagy and apoptosis, BNIP3 was significantly up-regulated within 48-72hrs post-infection. Of the genes linking autophagosomes to lysosomes, FAM176A was up-regulated throughout 24-72hrs post-infection. Conclusions: Modulation of autophagy and apoptosis genes occurs in neuronal cells at 24, 48, and 72hrs post- infection with Chlamydia pneumoniae. These genetic changes lead to dysfunction in these basic cellular processes; dysfunction in these processes has been shown to contribute to the neuropathology of late-onset Alzheimer’s disease. This work will allow future studies to further focus on the apoptotic and autophagic pathways to better understand how a pathogen such as Chlamydia pneumoniae plays a role in the development of late-onset Alzheimer’s disease.

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  Predictors of Successful Weight Loss Following Bariatric Surgery.

  F. R. Broder, Stacey C. Cahn, Robert A. DiTomasso, R. J. Petrucci, and P. S. Geno

  Poster presentation accepted for the 46th Annual Association for the Advancement of Behavioral and Cognitive Therapies, National Harbor, Maryland.

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  Reducing the Stigma of Mental Illness Among Medical Students

  Victoria Lawn, Matthew Jaffa, Catherine Babbitt-Cook, Burton Mark, Jane Dumsha, Marcus G. Bell, and Denah M. Appelt

  Abstract: The American Osteopathic Association House of Delegates Resolution 205 recommends “increased awareness of depression amongst U.S. Medical students” due to the increasing body of research describing the rise of depression, burn-out and suicide ideation among medical students. There is consequently a need to understand mental health issues as a component of professional development. Hypothesis: A student-led symposium addressing mental and emotional health topics relevant to medical students would reduce the stigma associated with mental illness. Materials and Methods: A 2-hour student-run “Patient Perspective” was held during the second neuroscience block at an osteopathic medical school in the northeastern United States. One week before the program, a student-developed online Wellness Survey measured prevalence of mental illness, common feelings during medical school, coping mechanisms used for stress, and use of mental health resources. Immediately before and after the program, students were asked to report their familiarity with mental illness and their feelings regarding a vignette about a mentally ill woman using “Mental Illness Among Us” pre and post surveys provided by the University of California San Francisco School of Medicine and adapted for the event. During the program, data from the online survey were shared, student organizers discussed emotional wellness and positive coping mechanisms in the context of the profession, and student panelists shared their experiences with mental health issues. A faculty psychiatrist spoke about mental health resources, and attendees received pamphlets listing these resources. The event concluded with student-led breakout sessions at which stress during medical school and strategies for promoting positive coping mechanisms were discussed, followed by the post survey. Results: 113 students completed the pre survey, 89 of whom completed the post survey. For these 89, differences between post and pre responses were universally in the direction of increasing acceptance and decreasing stigma of those with mental illness; all differences were statistically significant. The largest shift regarded students’ reluctance to disclose their own theoretical mental illness to colleagues. Conclusion: Incorporating an emotional health symposium into medical students’ training may increase understanding and acceptance of those who may have mental illness and reduce stigma associated with mental illness.

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  Insulin Pump School: Back to the Basics of Pump Therapy

  Sara Wilson Reece and Cheryl Williams

  The Longstreet Clinic (TLC), P.C. is a regional multidisciplinary physician practice. The Internal Medicine (IM) and Family Medicine (FM) departments are located in Gainesville and Oakwood (Hall County), Georgia. • Hall County is located in Northeast Georgia 50 miles northeast of Atlanta. It lies at the southern edge of the Chattahoochee National Forest and the foothills of the Blue Ridge Mountains. The population of Hall County is approximately 187,700. • Diabetes Education and Medical Nutritional Therapy department provides diabetes education and management services for IM and FM patients with diabetes. • Team of CDEs: RN, RD, PharmD • RN and PharmD are Certified Pump Trainers • Statistics • 20% of total IM and FM patients have diagnosis of diabetes. • 47% of patients with diabetes on insulin therapy. • 3% of patients on insulin therapy are on insulin pump therapy. • After a few insulin pump patients had severe hypoglycemic episodes while driving, the need for review of insulin pump survival skills was identified in order to prevent possible future incidents

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  Analysis of Chlamydia pneumoniae-infected monocytes following incubation with a novel peptide, acALY18, implicates the inflammasome in clearance of infection

  Brian J. Balin, James D. Thacker, Charles Lim, Corey M. Caruthers, Susan T. Hingley, Juliana Zoga, and Denah M. Appelt

  Chlamydia pneumoniae infection may be a trigger for the pathology observed in sporadic lateonset Alzheimer’s disease as a function of initiating neuroinflammation following entry of the organism into the brain. We have hypothesized that one entry mechanism may be by bloodborne infected monocytes trafficking the infection into the brain. This study focuses on infection of monocytes in vitro followed by analysis using immunofluorescence labeling and RT-PCR-microarray techniques. The microarrays utilized consisted of an Alzheimer’s disease pathway array and an innate and adaptive immunity array from SAbiosciences. Analysis by real time PCR for both gene arrays was performed on uninfected and C. pneumoniae-infected THP1 monocytes at 48 hr post-infection. In addition, we analyzed innate and adaptive immunity gene regulation changes following treatment of infected cells with a unique peptide, acALY18, derived from the endogenously expressed endoplasmic reticulum protein TRPC1. The peptide appears to stimulate the innate immune system through activation of the inflammasome. C. pneumoniae prominently infected THP1 monocytes at 24-48hr. Numerous large inclusions were identified using specific chlamydial monoclonal antibodies. Monocyte gene expression changes induced by infection with C. pneumoniae revealed significant up-regulation of 45 genes in the Alzheimer’s disease pathway. These included genes involved in: b-amyloid processing and clearance, apoptosis, proteases and protein kinases, and lipid metabolism. In contrast, infection resulted in significant changes in 30 genes governing innate and adaptive immunity including those for: the inflammatory response, host defense against bacteria, cytokines, chemokines, and an antibacterial humoral response. Intriguingly, following incubation of C. pneumoniae-infected cells with the acALY18 peptide (25-50nM) at 24hr post-infection, there was significant clearance of the organism from the monocytes as well as up-regulation of 38 genes. Our data suggest that C. pneumoniae infection of monocytes has a profound effect on gene regulation for both innate and adaptive immunity and for Alzheimer’s disease. Stimulating the innate immune response using the novel peptide, acALY18, promotes clearance of C. pneumoniae from infected monocytes; thereby implicating the inflammasome as a key component in eradicating this infection.

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  Analysis of Chlamydia pneumoniae and AD-like Pathology in the Brains of BALB/c Mice Following Direct Intra-cranial Infection

  Jessica Rachel Barton, Christine J. Hammond, Amy L. Brady, Denah M. Appelt, Brian J. Balin, and Christopher Scott Little

  Alzheimer’s disease (AD) is an age-related progressive neurodegenerative disorder and the most common form of dementia. The pathology in the central nervous system (CNS) impairs memory and cognition, hindering the capabilities and the quality of life of the individual. This project continues studying the role of infection and Alzheimer’s disease, as previous studies in this laboratory have done, and contributes to the overall understanding of the possible causes of this disease. In this study, BALB/c mice were infected, via direct intracranial injection, with a respiratory isolate (AR-39) of Chlamydia pneumoniae. Their brains were analyzed at 7 and 14 days post-infection, via immunohistochemistry, for the presence of C. pneumoniae, amyloid deposits and activated glial cells. The goal of this project was to measure the location and degree of C. pneumoniae burden, amyloid deposition and glial cell activation in the CNS following direct intracranial injection and to compare this data with results obtained from previous studies in this laboratory. We hypothesized that C. pneumoniae antigen and activated inflammatory cells will be observed in the infected mouse brains following direct intracranial injection and Aβ deposition will be observed in areas where inflammation occurs. C. pneumoniae, amyloid deposits and activated glial cells were detected in the brains following direct intracranial infection with C. pneumoniae. In infected mice there was an approximate 3.5-fold increase of C. pneumoniae antigen burden compared to uninfected mice at day 7 and there was an approximate 5.5-fold increase of C. pneumoniae antigen burden compared to uninfected mice at day 14. The burden of C. pneumoniae antigen, in the infected mice, increased 1.009-fold (no change) from day 7 to day 14 post-infection. The amyloid burden in infected mice increased approximately 3-fold compared to uninfected mice at day 7 and increased greater than10-fold compared to uninfected mice at day 14. The burden of amyloid, in the infected mice, increased 7-fold from day 7 to 14. From 7 to 14 days post-infection the C. pneumoniae and amyloid deposits located near the injection site spread distally from this location to other regions of the brain. Global activation of glia was observed in the CNS of infected mice at both 7 and 14 days post-infection. This data confirms that C. pneumoniae is capable of establishing an infection in the CNS. Although deposits were observed, the lack of a substantial amount of amyloid deposits suggested that the generation of deposits may require longer than 14 days following C. pneumoniae infection. As early as 7 days post-infection, inflammation is observed in response to the presence of C. pneumoniae and/or soluble amyloid in the CNS and the contribution of both infection with C. pneumoniae and the presence of soluble amyloid elicit the inflammatory response that presumably precedes and contributes to amyloid deposition

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  Infection of neuronal cells by Chlamydia pneumoniae and Herpes simplex virus type 1 alters expression of genes associated with Alzheimer’s disease

  Morgan M. Devins, Fiora D. Zoga, Brian J. Balin, Denah M. Appelt, and Susan T. Hingley

  Several studies have suggested an infectious etiology for Alzheimer’s disease (AD). We have been investigating a potential role for both Chlamydia pneumoniae and Herpes simplex virus type 1 (HSV1) in the initiation of sporadic late-onset AD. Our current study focuses on investigation of gene expression using Alzheimer-specific Real-Time PCR microarrays on RNA derived from SKNMC human neuronal cells infected with C. pneumoniae and/or HSV1. There are distinct differences in the patterns of gene regulation by the two pathogens. For example, C. pneumoniae induces expression of genes involved in amyloid production and processing, such as β-amyloid precursor protein (APP), β-site APP-cleaving enzyme 1 (BACE1), a γ-secretase complex protein (nicastrin [NCSTN]), NEDD8 activating enzyme E1 (NAE1), as well as a mitochondria-associated protein (hydroxysteroid (17-β) dehydrogenase 10 [HSD17B10]), α-2-macroglobulin (A2M) and the metallopeptidase ADAM9. Conversely, HSV1 tends to down-regulate expression of many genes, including those encoding a component of the γ-secretase complex (anterior pharynx defective 1 homolog A [APH1A]), low density lipoprotein related proteins (LRP1, LRP6, and LRP8), β-synuclein (SNCB) and ubiquinols (UQCRC1, UQCRC2). Co-infection with C. pneumoniae and HSV-1 produced a greater down-regulation of gene expression than that seen with HSV1 alone for several genes, including APP-like proteins (APLP1, APLP2) and kinases (cell division cycle 2 protein [CDC2], cyclin-dependent kinase [CDK5] and CDC2-related kinase [CDKL1]). Our data indicate that both C. pneumoniae and HSV1 can modulate expression of genes associated with AD, and thus could contribute to AD pathology, however these two pathogens likely act via different pathways. Furthermore, for several genes, co-infection with both C. pneumoniae and HSV1 appears to exacerbate the changes in gene expression seen with HSV1 alone.

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  Gestational Diabetes Clinic for Indigent Latinos

  Sara Wilson Reece

  Gestational diabetes mellitus (GDM), “carbohydrate intolerance of variable severity with onset or fist recognition during pregnancy,” results from insulin resistance and relative insulin deficiency usually in second trimester.¹ • Gestational diabetes (GDM) impacts between 4% and 9% of all pregnancies. • Gestational diabetes (GDM) results in increased fetal complications of macrosomia, shoulder dystocia and neonatal hyperglycemia as well as maternal risks of preeclampsia and polyhydramnois. • Women who are Hispanic or Asian decent are at highest risk of developing GDM. • Diagnosis ² • Perform 75-gramoral glucose tolerance test (OGTT) at 24 – 28 weeks of gestation in women not previously diagnosed with diabetes • Diagnosis of GDM made when any of the following values are exceeded: • Fasting ≥ 92 mg/dL • 1 hour ≥ 180 mg/dL • 2 hour ≥ 153 mg/dL • Upon diagnosis of gestational diabetes, medical nutrition therapy, self-monitoring of blood glucose and fetal monitoring are initiated. • Both Landon and Australian Carbohydrate Intolerance Study in Pregnant Women (ACHOIS) studies support active management of gestational diabetes even in the mild form to decrease fetal complications.

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  Chlamydophila (Chlamydia) pneumoniae promotes Ab 1-42 amyloid processing in Neuronal Cells: A Pathogenic Trigger for Alzheimer's Disease

  Elizabeth K. Ruszak, Karen C. McCarthy, Marcus G. Bell, Brian J. Balin, and Denah M. Appelt

  Background: Previously, our laboratory identified Chlamydophila (Chlamydia) pneumoniae (Cpn) in autopsied sporadic AD brains. Furthermore, we have developed a BALB/c mouse model that demonstrated infection-induced amyloid plaques similar to those found in AD, and demonstrated that Cpn infection of neuronal cells inhibited apoptotic pathways of cell death. Hypothesis: Our current studies address whether infection with Cpn in neuronal cells triggers abnormal cleavage of the beta amyloid precursor protein (bAPP) into Ab1-42, thereby contributing to amyloid plaque formation characteristic of the pathology identified in AD. Materials and Methods: Human neuroblastoma cells were infected with the respiratory strain AR39 Cpn in vitro, then amyloid processing was analyzed and quantitated using immunocytochemistry, Western blotting and ELISA assays. Results: Cpn was shown to infect neuronal cells and induce intracellular amyloid processing. Cpn infection yielded cytoplasmic labeling of Ab 1-42 that was increased relative to uninfected cells. The ELISA assay revealed that in neuronal cell lysates, Ab 1-42 in the infected cells was increased 3 to 16-fold over the uninfected cells, from 24 to 72hr post infection. Western blot analysis confirmed an increase in Ab 1-42 in the infected neuronal cell lysates. Conclusions: These data suggest that infection of neuronal cells with Chlamydophila (Chlamydia) pneumoniae alters the processing of bAPP, thereby producing Ab1-42. Therefore, these studies and previous research reported by our laboratory support the implication of Cpn as a pathogenic agent in perpetuating the hallmark amyloid plaque formations observed in AD. This concept holds major therapeutic considerations for future studies.

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  Herpes Simplex Virus 1 and Chlamydophila (Chlamydia) pneumoniae promote Ab 1-42 amyloid processing in murine astrocytes linking an infectious process to Alzheimer's disease

  Laura R. Triplett, Adam L. Dore, Kevin S. Kralik, Brian J. Balin, Susan T. Hingley, and Denah M. Appelt

  Background: Several studies have suggested an infectious etiology for Alzheimer's disease (AD). Previously, our laboratory identified Chlamydia pneumoniae (Cpn) from autopsied sporadic AD brains, as well as developed a BALB/c mouse model that demonstrated infection-induced amyloid plaques similar to those found in AD. Hypothesis: We propose that an additional pathogen such as herpes simplex virus type 1 (HSV1), also may be a contributing factor in toin the pathology seen in AD. HSV1, in addition to Cpn, may be triggering the abnormal cleavage of the beta amyloid precursor protein (bAPP) into Ab1-42 , thereby contributing to amyloid plaque formation. Our current study examines amyloid processing following infection of primary and C8-DIA murine astrocytes with Cpn and HSV1. Materials and Methods: Immunocytochemistry and western analysis was used to analyze the outcome of infection by these two pathogens. Results: Cpn infection resulted in an increase in cytoplasmic labeling of Ab 1-42 relative to uninfected cells, while increased nuclear labeling of Ab 1-42 was observed following HSV1 infection. Co-infections with Cpn and HSV1 resulted in amyloid labeling resembling that of HSV1 infection alone, though Ab 1-42 labeling appeared decreased specifically in Cpn-infected cells of the co-infected monolayers. Conclusions: These data suggest that infection of astrocytic cells by HSV1 and (Cpn) alter the processing of bAPP, thereby producing Ab1-42. Therefore, these studies, inaddition to the previous research reported by our laboratory, support an emerging linkage of the infectious processs to the neuropathology characteristic of Alzheimer's disease.