Investigation of the P13K/AKT/FOXO and the Ras/Raf/MEK/ERK Signalling Pathways in Ovarian Development

The formation and development of ovarian follicles is regulated by the interactions of germ cells and somatic cells, intraovarian growth regulatory factors and the pituitary gonadotrophins, FSH and LH. Despite the fact that granulosa cells of large follicles are highly proliferative, granulosa cell tumours are rare suggesting that potent regulatory mechanisms, including apoptotic factors, control proliferation and differentiation of these cells. Two critical pathways that intercommunicate to influence ovarian cell proliferation and differentiation are the PI3K1AKT/FOXOI pathway and the Ras/Raf/MEKIERK cascade. As FSH can stimulate both pathways, a super-ovulatory regimen of hormones PMSG/hCG (FSHlLH analogues) administered to immature wildtype mice allowed the investigation of key components in these pathways at different stages of follicular development. In this project cell cycle regulators and apoptotic markers were investigated to determine the role they play in ovarian development. Specific PI3K and Ras pathway components were studied to decipher how they behaved throughout follicular growth. Results indicated that the exact phosphorylation and expression pattern of proteins from these pathways happens in a very transient, coordinated and timely way. Granulosa cell culture work displayed that extracellular ligands as well as FSH have the ability to stimulate both the PI3K pathway and the Ras pathway in quite a time-dependent manner. The transcription factor CEBPP is present from once hCG is administered to 5 days post hCG treatment suggesting it plays a role at many stages of follicular development as well as at the corpora lute a regression stage. Some mutant mouse models were investigated to detennine the importance of key components in the PI3K pathway and the Ras pathway. This novel insight into normal ovarian development suggests that it is the specific time dependent expression and cross talk of these pathway components that ultimately results in the coordinated and synchronised growth of immature follicles to the corpora lute a stage

Antiviral TRIMs: Friend or Foe in Autoimmune and Autoinflammatory Disease?

The concept that viral sensing systems, via their ability to drive pro-inflammatory cytokine and interferon production, contribute to the development of autoimmune and autoinflammatory disease is supported by a wide range of clinical and experimental observations. Recently, the tripartite motif-containing proteins (TRIMs) have emerged as having key roles in antiviral immunity — either as viral restriction factors or as regulators of pathways downstream of viral RNA and DNA sensors, and the inflammasome. Given their involvement in these pathways, we propose that TRIM proteins contribute to the development and pathology of autoimmune and autoinflammatory conditions, thus making them potential novel targets for therapeutic manipulation

Archeota, Spring/Summer 2022

Archeota is a platform for SJSU iSchool students to contribute to the archival conversation. It is written BY students, FOR students. It provides substantive content on archival concerns and issues and promotes professional development in the field of archival studies. Archeota upholds the core values of the archival profession. Contents: From Ashes to Archive: Photojournalist Elizabeth Sunflower’s Body of Work By Laura Darlington Accessibility in Archival Spaces: Breaking Down Barriers for Archival Workers with Disabilities By Kate Goodwin Queer Zine Archive Project: Building a Community Archive of Living History By Alice Wynne Brewster Kahle’s Vision for the Future of Libraries Interview with the Founder of the Internet Archive By Claire Kelley Text, Prose & RocknRoll Podcast: Preserving the Diverse History of Popular Music By Sharon Kosach Farewell to Our Spring 2022 Graduates Interviews With SAA Student Chapter Leaders A Jew in a Catholic Domain: Internship at Schools of the Sacred Heart San Francisco By Max Rosen Summer Reading Recommendations Time to Curl Up With a Good Book! Past Event Archive SJSU SAA Student Chapter events AY 2021-2022https://scholarworks.sjsu.edu/saasc_archeota/1015/thumbnail.jp

Discrimination of immune cell activation using Ramanmicro-spectroscopy in anin-vitro & ex-vivo model

Activation and proliferation of immune cells such as lymphocytes and monocytes are appropriate inflammatory responses to invading pathogens and are key to overcoming an infection. In contrast, uncontrolled and prolonged activation of these cellular signalling pathways can be deleterious to the body and result in the development of autoimmune conditions. The understanding of cellular activatory status therefore plays a significant role in disease diagnosis and progression. Conventional automated approaches such as enzyme linked immunosorbent assays (ELISA) and immune-labelling techniques are time-consuming and expensive, relying on a commercially available and specific antibody to identify cell activation. Developing a label-free method for assessing molecular changes would therefore offer a quick and cost-efficient alternative in biomedical research. Here Raman spectroscopy is presented as an effective spectroscopic method for the identification of activated immune cells using both cell lines and primary cells (including purified monocyte and lymphocyte subgroups and mixed peripheral blood mononuclear cell (PBMC) populations) obtained from healthy donors. All cell lines and primary cells were exposed to different stimulants and cellular responses confirmed by flow cytometry or ELISA. Machine learning models of cell discrimination using Raman spectra were developed and compared to reference flow-cytometry, with spectral discrimination levels comparing favourably with the reference method. Spectral signatures of molecular expression after activation were also extracted with results demonstrating alignment with expected profiles. High performance classification models constructed in these in-vitro and ex-vivo studies enabled identification of the spectroscopic discrimination of immune cell subtypes in their resting and activated state. Further spectral fitting analysis identified a number of potential spectral biomarkers that elucidate the spectral classification

A Novel Pool of Microparticle Cholesterol Is Elevatedin Rheumatoid Arthritis but Not in Systemic Lupus Erythematosus Patients

Microparticles are sub-micron, membrane-bound particles released from virtually allcells and which are present in the circulation. In several autoimmune disorders their amountand composition in the circulation is altered. Microparticle surface protein expression has beenexplored as a differentiating tool in autoimmune disorders where the clinical pictures can overlap.Here, we examine the utility of a novel lipid-based marker—microparticle cholesterol, present in allmicroparticles regardless of cellular origin—to distinguish between rheumatoid arthritis (RA) andsystemic lupus erythematosus (SLE). We first isolated a series of microparticle containing lipoproteindeficient fractions from patient and control plasma. There were no significant differences in thesize, structure or protein content of microparticles isolated from each group. Compared to controls,both patient groups contained significantly greater amounts of platelet and endothelial cell-derivedmicroparticles. The cholesterol content of microparticle fractions isolated from RA patients wassignificantly greater than those from either SLE patients or healthy controls. Our data indicate thatcirculating non-lipoprotein microparticle cholesterol, which may account for 1–2% of measuredcholesterol in patient samples, may represent a novel differentiator of disease, which is independentof cellular origi

Herpes Simplex Virus 1 Targets IRF7 via ICP0 to Limit Type I IFN Induction

Herpes simplex keratitis (HSK), caused by herpes simplex virus type 1 (HSV‑1) infection, is the commonest cause of infectious blindness in the developed world. Following infection the virus is initially suspended in the tear film, where it encounters a multi‑pronged immune response comprising enzymes, complement, immunoglobulins and crucially, a range of anti‑viral and pro‑inflammatory cytokines. However, given that HSV‑1 can overcome innate immune responses to establish lifelong latency throughout a susceptible individual’s lifetime, there is significant interest in understanding the mechanisms employed by HSV‑1 to downregulate the anti‑viral type I interferon (IFN) mediated immune responses. This study aimed to investigate the interactions between infected cell protein (ICP)0 and key elements of the IFN pathway to identify possible novel targets that contribute to viral immune evasion. Reporter gene assays demonstrated the ability of ICP0 to inhibit type I IFN activity downstream of pathogen recognition receptors (PRRs) which are known to be involved in host antiviral defences. Further experiments identified interferon regulatory factor (IRF)7, a driver of type I IFN, as a potential target for ICP0. These findings increase our understanding of the pathogenesis of HSK and suggest IRF7 as a potential therapeutic target

Btk Regulated Macrophage Polarization in Response to Lipopolysaccharide

Bacterial Lipopolysaccharide (LPS) is a strong inducer of inflammation and does so by inducing polarization of macrophages to the classic inflammatory M1 population. Given the role of Btk as a critical signal transducer downstream of TLR4, we investigated its role in M1/M2 induction. In Btk deficient (Btk−\−) mice we observed markedly reduced recruitment of M1 macrophages following intraperitoneal administration of LPS. Ex vivo analysis demonstrated an impaired ability of Btk−/− macrophages to polarize into M1 macrophages, instead showing enhanced induction of immunosuppressive M2-associated markers in response to M1 polarizing stimuli, a finding accompanied by reduced phosphorylation of STAT1 and enhanced STAT6 phosphorylation. In addition to STAT activation, M1 and M2 polarizing signals modulate the expression of inflammatory genes via differential activation of transcription factors and regulatory proteins, including NF-κB and SHIP1. In keeping with a critical role for Btk in macrophage polarization, we observed reduced levels of NF-κB p65 and Akt phosphorylation, as well as reduced induction of the M1 associated marker iNOS in Btk−/− macrophages in response to M1 polarizing stimuli. Additionally enhanced expression of SHIP1, a key negative regulator of macrophage polarisation, was observed in Btk−/− macrophages in response to M2 polarizing stimuli. Employing classic models of allergic M2 inflammation, treatment of Btk−/− mice with either Schistosoma mansoni eggs or chitin resulted in increased recruitment of M2 macrophages and induction of M2-associated genes. This demonstrates an enhanced M2 skew in the absence of Btk, thus promoting the development of allergic inflammation

Higher Education Exchange: 2012

This annual publication serves as a forum for new ideas and dialogue between scholars and the larger public. Essays explore ways that students, administrators, and faculty can initiate and sustain an ongoing conversation about the public life they share.The Higher Education Exchange is founded on a thought articulated by Thomas Jefferson in 1820: "I know no safe depository of the ultimate powers of the society but the people themselves; and if we think them not enlightened enough to exercise their control with a wholesome discretion, the remedy is not to take it from them, but to inform their discretion by education."In the tradition of Jefferson, the Higher Education Exchange agrees that a central goal of higher education is to help make democracy possible by preparing citizens for public life. The Higher Education Exchange is part of a movement to strengthen higher education's democratic mission and foster a more democratic culture throughout American society.Working in this tradition, the Higher Education Exchange publishes interviews, case studies, analyses, news, and ideas about efforts within higher education to develop more democratic societies

Genetics of SLE: Functional Relevance for Monocytes/Macrophages in Disease

Genetic studies in the last 5 years have greatly facilitated our understanding of how the dysregulation of diverse components of the innate immune system contributes to pathophysiology of SLE. A role for macrophages in the pathogenesis of SLE was first proposed as early as the 1980s following the discovery that SLE macrophages were defective in their ability to clear apoptotic cell debris, thus prolonging exposure of potential autoantigens to the adaptive immune response. More recently, there is an emerging appreciation of the contribution both monocytes and macrophages play in orchestrating immune responses with perturbations in their activation or regulation leading to immune dysregulation. This paper will focus on understanding the relevance of genes identified as being associated with innate immune function of monocytes and macrophages and development of SLE, particularly with respect to their role in (1) immune complex (IC) recognition and clearance, (2) nucleic acid recognition via toll-like receptors (TLRs) and downstream signalling, and (3) interferon signalling. Particular attention will be paid to the functional consequences these genetic associations have for disease susceptibility or pathogenesis