Development of an Enzyme-Linked Immunosorbent Assay (ELISA) for the Quantification of ARID1A in Tissue Lysates

ARID1A is a subunit of the mammalian SWI/SNF complex, which is thought to regulate gene expression through restructuring chromatin structures. Its gene ARID1A is frequently mutated and ARID1A levels are lowered in several human cancers, especially gynecologic ones. A functional ARID1A loss may have prognostic or predictive value in terms of therapeutic strategies but has not been proposed based on a quantitative method. Hardly any literature is available on ARID1A levels in tumor samples. We developed an indirect enzyme-linked immunosorbent assay (ELISA) for ARID1A based on the current EMA and FDA criteria. We demonstrated that our ELISA provides the objective, accurate, and precise quantification of ARID1A concentrations in recombinant protein solutions, cell culture standards, and tissue lysates of tumors. A standard curve analysis yielded a ‘goodness of fit’ of R2 = 0.99. Standards measured on several plates and days achieved an inter-assay accuracy of 90.26% and an inter-assay precision with a coefficient of variation of 4.53%. When tumor lysates were prepared and measured multiple times, our method had an inter-assay precision with a coefficient of variation of 11.78%. We believe that our suggested method ensures a high reproducibility and can be used for a high sample throughput to determine the ARID1A concentration in different tumor entities. The application of our ELISA on various tumor and control tissues will allow us to explore whether quantitative ARID1A measurements in tumor samples are of predictive value

Intracranial Intracerebral Schwannoma: a Case Report and Review of the Literature

Intracranial schwannomas are relatively uncommon, accounting for approximately 8% of all intracranial tumors, while intracerebral schwannomas represent an even rarer entity, responsible for roughly 1% of all intracranial schwannomas. After reviewing the relevant literature, we discussed the clinical journey of a 74-year-old woman who presented with a 3-week history of dizziness and nausea. Magnetic resonance imaging revealed a right temporal mass lesion with perifocal edema. The initial suspicion was the diagnosis of a glioblastoma or metastasis, prompting surgical intervention. During the surgery, a gross total resection of a noninvasive tumor was successfully performed. The patient’s postoperative recovery was uneventful. Histopathological examination and confrmatory immunohistochemistry played a crucial role in reaching the fnal diagnosis of an intracerebral temporal schwannoma, highlighting the diagnostic challenges posed by radiologically indistinguishable features from metastasis and gliomas. Despite these challenges, complete surgical removal remains the most preferred treatment option, resulting in a favorable long-term prognosis without the need for adjuvant or neoadjuvant chemotherapy. Intracerebral schwannomas are exceedingly rare brain tumors, often found on the brain’s surface or adjacent ventricles. Early and accurate diagnosis can be challenging due to radiological features overlapping with other intracranial pathologies. Nonetheless, histopathological examination and immunohistochemistry remain indispensable tools in establishing a defnitive diagnosis and guiding efective treatment strategies. With complete surgical excision, patients with intracerebral schwannomas can expect a positive outcome and a promising long-term prognosis. Further research and case studies are warranted to enhance our understanding of these rare tumors and improve patient outcomes

Bridging the rodent to human translational gap: Marmosets as model systems for the study of Alzheimer\u27s disease.

INTRODUCTION: Our limited understanding of the mechanisms that trigger the emergence of Alzheimer\u27s disease (AD) has contributed to the lack of interventions that stop, prevent, or fully treat this disease. We believe that the development of a non-human primate model of AD will be an essential step toward overcoming limitations of other model systems and is crucial for investigating primate-specific mechanisms underlying the cellular and molecular root causes of the pathogenesis and progression of AD. METHODS: A new consortium has been established with funding support from the National Institute on Aging aimed at the generation, characterization, and validation of Marmosets As Research Models of AD (MARMO-AD). This consortium will study gene-edited marmoset models carrying genetic risk for AD and wild-type genetically diverse aging marmosets from birth throughout their lifespan, using non-invasive longitudinal assessments. These include characterizing the genetic, molecular, functional, behavioral, cognitive, and pathological features of aging and AD. RESULTS: The consortium successfully generated viable founders carrying DISCUSSION: By establishing marmoset models of AD, we will be able to investigate primate-specific cellular and molecular root causes that underlie the pathogenesis and progression of AD, overcome limitations of other model organisms, and support future translational studies to accelerate the pace of bringing therapies to patients

Permafrost Active Layer Microbes From Ny Ålesund, Svalbard (79°N) Show Autotrophic and Heterotrophic Metabolisms With Diverse Carbon-Degrading Enzymes

The active layer of permafrost in Ny Ålesund, Svalbard (79°N) around the Bayelva River in the Leirhaugen glacier moraine is measured as a small net carbon sink at the brink of becoming a carbon source. In many permafrost-dominating ecosystems, microbes in the active layers have been shown to drive organic matter degradation and greenhouse gas production, creating positive feedback on climate change. However, the microbial metabolisms linking the environmental geochemical processes and the populations that perform them have not been fully characterized. In this paper, we present geochemical, enzymatic, and isotopic data paired with 10 Pseudomonas sp. cultures and metagenomic libraries of two active layer soil cores (BPF1 and BPF2) from Ny Ålesund, Svalbard, (79°N). Relative to BPF1, BPF2 had statistically higher C/N ratios (15 ± 1 for BPF1 vs. 29 ± 10 for BPF2; n = 30, p < 10–5), statistically lower organic carbon (2% ± 0.6% for BPF1 vs. 1.6% ± 0.4% for BPF2, p < 0.02), statistically lower nitrogen (0.1% ± 0.03% for BPF1 vs. 0.07% ± 0.02% for BPF2, p < 10–6). The d13C values for inorganic carbon did not correlate with those of organic carbon in BPF2, suggesting lower heterotrophic respiration. An increase in the δ13C of inorganic carbon with depth either reflects an autotrophic signal or mixing between a heterotrophic source at the surface and a lithotrophic source at depth. Potential enzyme activity of xylosidase and N-acetyl-β-D-glucosaminidase increases twofold at 15°C, relative to 25°C, indicating cold adaptation in the cultures and bulk soil. Potential enzyme activity of leucine aminopeptidase across soils and cultures was two orders of magnitude higher than other tested enzymes, implying that organisms use leucine as a nitrogen and carbon source in this nutrient-limited environment. Besides demonstrating large variability in carbon compositions of permafrost active layer soils only ∼84 m apart, results suggest that the Svalbard active layer microbes are often limited by organic carbon or nitrogen availability and have adaptations to the current environment, and metabolic flexibility to adapt to the warming climate.Peer Reviewe

The Grizzly, April 2, 2009

Monologues Give Vaginas Voice • Smelly, Spicy, Stimulating Aphrodisiacs: Fact or Fiction? • 26th Annual Airband • Spring Break Fun in the Sun with Habitat for Humanity • TA Eliana Henriquez • Restaurant Reviews: Irish Joe\u27s Cafe; Rock Bottom Brewery • Opinions: George W. Bush Legacy; Letter from a Local Resident • Pre-Season Update: UC Men\u27s Golf Gears Up for a New Seasonhttps://digitalcommons.ursinus.edu/grizzlynews/1784/thumbnail.jp

Bisulfite profiling of the MGMT promoter and comparison with routine testing in glioblastoma diagnostics

Background: Promoter methylation of the DNA repair gene O6 -methylguanine-DNA methyltransferase (MGMT) is an acknowledged predictive epigenetic marker in glioblastoma multiforme and anaplastic astrocytoma. Patients with methylated CpGs in the MGMT promoter beneft from treatment with alkylating agents, such as temozolomide, and show an improved overall survival and progression-free interval. A precise determination of MGMT promoter methyla‑ tion is of importance for diagnostic decisions. We experienced that diferent methods show partially divergent results in a daily routine. For an integrated neuropathological diagnosis of malignant gliomas, we therefore currently apply a combination of methylation-specifc PCR assays and pyrosequencing. Results: To better rationalize the variation across assays, we compared these standard techniques and assays to deep bisulfte sequencing results in a cohort of 80 malignant astrocytomas. Our deep analysis covers 49 CpG sites of the expanded MGMT promoter, including exon 1, parts of intron 1 and a region upstream of the transcription start site (TSS). We observed that deep sequencing data are in general in agreement with CpG-specifc pyrosequencing, while the most widely used MSP assays published by Esteller et al. (N Engl J Med 343(19):1350–1354, 2000. https://doi.org/ 10.1056/NEJM200011093431901) and Felsberg et al. (Clin Cancer Res 15(21):6683–6693, 2009. https://doi.org/10.1158/ 1078-0432.CCR-08-2801) resulted in partially discordant results in 22 tumors (27.5%). Local deep bisulfte sequencing (LDBS) revealed that CpGs located in exon 1 are suited best to discriminate methylated from unmethylated samples. Based on LDBS data, we propose an optimized MSP primer pair with 83% and 85% concordance to pyrosequencing and LDBS data. A hitherto neglected region upstream of the TSS, with an overall higher methylation compared to exon 1 and intron 1 of MGMT, is also able to discriminate the methylation status. Conclusion: Our integrated analysis allows to evaluate and redefne co-methylation domains within the MGMT pro‑ moter and to rationalize the practical impact on assays used in daily routine diagnostics

The Grizzly, November 13, 2008

Berman Expands to Envelop Art and Imagination • Election Night Brings Cheer to Liberal Ursinus College • Changes Expected for This Year\u27s New Member Education • Passage of Prop 8 Leaves Same-Sex Couples Concerned • Days After Election: Obama Gets to Work • Forensics: Not the Science, but the Ursinus Debate Team • Escape Velocity\u27s Eternalmotion Strikes a Chord at UC • Finding the Phillies Parade: An Unforgettable Adventure • History Attached to a Philly Row Home • Opinions: Barack Obama: Our Next President • Kait Sutherland Finishes Out Hockey Reign at Ursinushttps://digitalcommons.ursinus.edu/grizzlynews/1775/thumbnail.jp

The Grizzly, September 11, 2008

Republican National Convention Rallies Supporters • Studies Show Half of College Students Contemplate Suicide • Start of New Now on View Art Film Festival • Science, Strife and Speculation of Hurricane Season \u2708 • UC Senior Implements First Ever Bike Share Program • Cinematic Guild Brings Creative Filmography to Campus • Introducing Cassandra Aguilar • The Grizzly Catches Up with Dean Nolan • Celebrating Herencia Latina Fall Events • Molly Maguire\u27s: A Bit O\u27 Irish Spirit • UC Women\u27s Soccer Fights for Another Strong Season • Football\u27s New Motto: No Interceptions, No Exceptionshttps://digitalcommons.ursinus.edu/grizzlynews/1767/thumbnail.jp

The Grizzly, September 4, 2008

Democrats Deem 2008 Election A Fight for the Future • Current Look at Issues Surrounding Iraqi Sovereignty • Changing Face of Ursinus: Recent Renovations • Ursinus College Journeys Abroad: Tuebingen, Germany • Another Season of Reviewing Begins with a Shining Culinary Success • Incredible Medical Internship for Senior • UC Men\u27s Soccer Team Intends on Silencing the Criticshttps://digitalcommons.ursinus.edu/grizzlynews/1766/thumbnail.jp

Regulation of nerve growth and patterning by cell surface protein disulphide isomerase.

Contact repulsion of growing axons is an essential mechanism for spinal nerve patterning. In birds and mammals the embryonic somites generate a linear series of impenetrable barriers, forcing axon growth cones to traverse one half of each somite as they extend towards their body targets. This study shows that protein disulphide isomerase provides a key component of these barriers, mediating contact repulsion at the cell surface in chick half-somites. Repulsion is reduced both in vivo and in vitro by a range of methods that inhibit enzyme activity. The activity is critical in initiating a nitric oxide/S-nitrosylation-dependent signal transduction pathway that regulates the growth cone cytoskeleton. Rat forebrain grey matter extracts contain a similar activity, and the enzyme is expressed at the surface of cultured human astrocytic cells and rat cortical astrocytes. We suggest this system is co-opted in the brain to counteract and regulate aberrant nerve terminal growth