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

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.https://digitalcommons.pcom.edu/posters/1007/thumbnail.jp

Anterior Talofibular Ligament Abnormality on Routine Magnetic Resonance Imaging of the Ankle

The anterior talo­fibular ligament (ATFL) extends from the anteroinferior border of the ­fibula to the talar neck. Primary restraint to ankle inversion in plantar‑exion. Injury (acute or chronic) can be diagnosed with physical exam, stress X-Rays, ultrasound or magnetic resonance imaging (MRI). Purpose: MRI abnormalities in asymptomatic individuals known in other areas of orthopaedics (shoulder and spine). Purpose of our study: determine the prevalence of ATFL abnormalities found on MRI in asymptomatic individuals. Asymptomatic individuals - those undergoing MRI for pathology unrelated to lateral ankle trauma, instability, or inversion injuries

Infection with Chlamydia Pneumoniae Alters Calcium-associated Gene Regulation and Processes in Neuronal Cells and Monocytes: Implications for Alzheimer’s Disease

Background: First proposed by Khachaturian in 1994, the calcium hypothesis postulates that sustained disturbance of intracellular calcium is the leading cause of neurodegenerative disorders. Studies showing alteration in calcium signaling in both sporadic and familial Alzheimer’s disease (AD) support this hypothesis. Intracellular calcium signaling is tightly regulated in time, intensity, and space, and is responsible for a variety of neuronal functions. Calcium influx from the extracellular environment modulates calcium levels, as do intracellular stores in the endoplasmic reticulum. The focus of this study was to test various calcium related genes in both monocytes and neuronal cells. Previous studies have shown that cells infected with Chlamydia pneumoniae (Cpn) exhibit altered protein processing, such as amyloid and tau modification, consistent with those found in AD. We expect to see significant alterations in calcium genes, as well as their protein products in Cpn infected cells. Every calcium gene has a unique function in the cell. Determining which genes are up or down regulated following infection may provide insight into how the neurodegeneration process observed in AD is initiated by Cpn infections

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

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.https://digitalcommons.pcom.edu/posters/1005/thumbnail.jp

Sistem Pengelolaan Cashflow Keuangan Online Di Pabrik Usantex Surakarta

Today, many companies are utilizing information systems in managing business activity data. One of them is the Usantex Surakarta Factory, which already uses an online system for managing cashflow data. However, in some parts the system still has shortcomings. First, in the cashflow menu section which only counts 50 cash in and out data that appears on the page, it should count along with previous transactions. Then, the debt data that is still not displayed based on the debtor. Therefore, in this final project develops a new system using the PHP programming language and PhpMyAdmin database system. This new online system will solve the problems in the previous system as mentioned above. System development method uses Waterfall, which consists of 5 stages including requirements analysis, system design, writing program code, testing, and implementation and maintenance. The new online system testing uses the black box method to ensure all system functions are running as expected and testing the user to find out the suitability of the system with Usantex through a questionnaire using the likert scale calculation. Based on the test results it can be concluded that the system runs according to its function and the percentage of the results of the questionnaire gets an average score of 92.5% so that it is in accordance with the request from Usantex

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

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.https://digitalcommons.pcom.edu/posters/1009/thumbnail.jp

Changes in Expression of Genes Associated with Autophagy and Apoptosis in Neuronal Cells Infected with HSV-1may Suggest Infection-induced Mechanisms of Neurodegeneration

Background:This study investigates the potential role of herpes simplex virus type 1 (HSV-1) in the pathogenesis of neurodegenerative disorders, such as Alzheimer’s disease (AD), by exploring changes in gene expression related to antiviral immunity and the autophagic pathway. Autophagy is a process that recycles organelles and proteins to create more energy for the cell. This pathway has been linked to neurodegeneration, as malfunctions in the completion of this process lead to a decline in overall cellular health and function. Interestingly, HSV-1 has been shown to block the completion of autophagy, which would potentially contribute to the cytopathic changes observed in AD

Bone stress injury of the ankle in professional ballet dancers seen on MRI

<p>Abstract</p> <p>Background</p> <p>Ballet Dancers have been shown to have a relatively high incidence of stress fractures of the foot and ankle. It was our objective to examine MR imaging patterns of bone marrow edema (BME) in the ankles of high performance professional ballet dancers, to evaluate clinical relevance.</p> <p>Methods</p> <p>MR Imaging was performed on 12 ankles of 11 active professional ballet dancers (6 female, 5 male; mean age 24 years, range 19 to 32). Individuals were imaged on a 0.2 T or 1.5 T MRI units. Images were evaluated by two musculoskeletal radiologists and one orthopaedic surgeon in consensus for location and pattern of bone marrow edema. In order to control for recognized sources of bone marrow edema, images were also reviewed for presence of osseous, ligamentous, tendinous and cartilage injuries. Statistical analysis was performed to assess the strength of the correlation between bone marrow edema and ankle pain.</p> <p>Results</p> <p>Bone marrow edema was seen only in the talus, and was a common finding, observed in nine of the twelve ankles imaged (75%) and was associated with pain in all cases. On fluid-sensitive sequences, bone marrow edema was ill-defined and centered in the talar neck or body, although in three cases it extended to the talar dome. No apparent gender predilection was noted. No occult stress fracture could be diagnosed. A moderately strong correlation (phi = 0.77, p= 0.0054) was found between edema and pain in the study population.</p> <p>Conclusion</p> <p>Bone marrow edema seems to be a specific MRI finding in the talus of professional ballet dancers, likely related to biomechanical stress reactions, due to their frequently performed unique maneuvers. Clinically, this condition may indicate a sign of a bone stress injury of the ankle.</p

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

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.https://digitalcommons.pcom.edu/posters/1001/thumbnail.jp

'Treatment of the Sportsman's groin': British Hernia Society's 2014 position statement based on the Manchester Consensus Conference

&lt;b&gt;Introduction&lt;/b&gt; The aim was to produce a multidisciplinary consensus to determine the current position on the nomenclature, definition, diagnosis, imaging modalities and management of Sportsman's groin (SG).&lt;p&gt;&lt;/p&gt; &lt;b&gt;Methods&lt;/b&gt; Experts in the diagnosis and management of SG were invited to participate in a consensus conference held by the British Hernia Society in Manchester, UK on 11–12 October 2012. Experts included a physiotherapist, a musculoskeletal radiologist and surgeons with a proven track record of expertise in this field. Presentations detailing scientific as well as outcome data from their own experiences were given. Records were made of the presentations with specific areas debated openly.&lt;p&gt;&lt;/p&gt; &lt;b&gt;Results&lt;/b&gt; The term ‘inguinal disruption’ (ID) was agreed as the preferred nomenclature with the term ‘Sportsman's hernia’ or ‘groin’ rejected, as no true hernia exists. There was an overwhelming agreement of opinion that there was abnormal tension in the groin, particularly around the inguinal ligament attachment. Other common findings included the possibility of external oblique disruption with consequent small tears noted as well as some oedema of the tissues. A multidisciplinary approach with tailored physiotherapy as the initial treatment was recommended with any surgery involving releasing the tension in the inguinal canal by various techniques and reinforcing it with a mesh or suture repair. A national registry should be developed for all athletes undergoing surgery.&lt;p&gt;&lt;/p&gt; &lt;b&gt;Conclusions&lt;/b&gt; ID is a common condition where no true hernia exists. It should be managed through a multidisciplinary approach to ensure consistent standards and outcomes are achieved