bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Bcl‐2 in Hematological Cancer Formation
Authors: Link Tina , Fortier Abigail , Jain Supriya , Kepner Alisa , Koh Ally , Stanovski Lisa , Ul‐Islam Misha , Walter Ellie 
Published: November 2023

B cell leukemia/lymphoma 2 (BCL‐2) is a 26 kilodalton, amphipathic, anti‐apoptotic protein located on the outer mitochondrial membrane. BCL‐2 interacts with other anti‐apoptotic and pro‐apoptotic factors in order to regulate apoptosis. If a structural change occurs on the BCL‐2 gene, overexpression of the BCL‐2 protein can result. This causes a change in the ratio of BCL‐2 to its BCL‐2 Homologous 3 (BH3) ligands, preventing the release of the apoptotic signal. The primary change associated with BCL‐2 overexpression in lymphomas is a t(14;18) chromosomal translocation, which causes an increase in BCL‐2 transcription. The upregulation of the oncogene allows for the survival of the mutated B cells by raising the apoptotic threshold, which then dislodges regular lymphocytes in the bone marrow or lymph nodes. Lack of cell death due to overexpression of BCL‐2 has been linked to hematological malignancies, specifically lymphomas and myelomas. Both B‐lymphocytic cancers can result from alterations of the immunoglobulin heavy chain locus on chromosome 14. While BCL2 translocation can cause Follicular Lymphoma, BCL2 over‐expression is observed in many other tumor types and likely contributes to cancer progression and drug resistance in these tumors. Treatments to inhibit BCL‐2 include attempts at activation and overexpression of the pro apoptotic BH3 proteins, BCL‐2 Associated X Protein (BAX) and BCL‐2 Associated K protein (BAK), and creation of BH3 mimetics. Indeed, the FDA approved the BCL2 inhibitor Venetoclax (ABT‐199) earlier this year for the treatment of Chronic Lymphocytic Leukemia. These treatments could help cause cancer cell death by binding to targeted anti‐apoptotic proteins and causing the signal for apoptosis to be released. This could neutralize the cancerous effects of the BCL‐2 oncogene upregulation. Walton High School SMART (Students Modeling A Research Topic) Team used the computer program Jmol to create a 3D printed model of BCL‐2 and composed a review poster of our secondary research to investigate and demonstrate the relationship between the structure and function of this molecule.Support or Funding InformationDiane Munzenmaier, Ph.D., Center for Biomolecular Modeling, Milwaukee School of Engineering
CINOBUFOTALIN INHIBITS OVARIAN CANCER CELL METASTASIS VIA APOPTOTIC SIGNALING AND TARGETING THE mTOR PATHWAY
Authors: Afroze Syeda Humayra , Peddaboina Chander , Dean DeShaundra M , McDowell Anthony , McCormick Timothy C , Newell‐Rogers Karen , Zawieja David C , Kuehl Thomas J , Uddin Mohammad N 
Published: November 2023

ObjectiveCinobufotalin (CINO), a cardiotonic steroid (CTS) or bufadienolide, is extracted from the skin secretions of the traditional Chinese medicine giant toads (Chan su). CINO has been used as a cardiotonic, diuretic and a hemostatic agent. Previously we have shown that CINO inhibits the cytotrophoblast cell function. Recently other study has shown that CINO inhibits A549, a lung cancer cell function. We demonstrated that CINO inhibited the proliferation, migration and invasion of three ovarian cancer cell lines; SK‐OV‐3, CRL‐1978 and CRL‐11731. In this study we evaluated the cellular mechanisms by which CINO inhibits ovarian cancer cell function.Study DesignWe evaluated the effect of CINO on three ovarian cancer cells SK‐OV‐3, CRL‐1978, and CRL‐11731 function in vitro. Each Cell lines were treated with different concentrations of CINO (0.1, 1, 5 and 10 μM). For each cell line cell proliferation, migration and invasion were measured by using a CellTiter Assay (Promega), Cytoselect Assay (Cell Biolabs) and by using a FluoroBlock Assay (BD) respectively. Proliferating Cell Nuclear Antigen (PCNA), mTOR and pAKT were also evaluated in cell lysates of CINO treated these 3 ovarian cancer cells by western blot analysis. Cell Cycle arrest and Cell viability were determined by fluorescence‐activated cell sorting (FACS) analysis. We also performed Annexin V staining on CINO treated these 3 ovarian cancer cell lines by immunofluorescence to evaluate the pro‐apoptotic protein expression. In addition mitochondrial membrane potential (MMP) has also been measured for all these 3 ovarian cell lines after CINO treatment using MMP kit, by FACS analysis.ResultsConcentration of CINO at 0.5μM inhibit SK‐OV‐3, CRL‐1978, and CRL‐11731 ovarian cancer cells proliferation, migration and invasion without cell death and loss of cell viability but cell viability differs for each cell line. Each cell lines differ in response to CINO doses for PCNA expression as well as Annexin V pro‐apoptotic protein expression. CINO decreases mitochondrial membrane potential for SK‐OV‐3 but not for CRL‐1978 and CRL‐11731 increases in response to CINO treatment. CINO causes the decrease in mTOR and pAKT expression in SK‐OV‐3 and CRL‐1978 but not in CRL‐11731.ConclusionCINO is cell specific, as each cancer cell line responds differently. These data demonstrate that the mode of action of CINO is different on these 3 types of ovarian cancer cells. Moreover, it also can be suggested that CINO inhibits the ovarian cancer cell metastasis via apoptotic signaling and targeting the mTOR pathway.Support or Funding InformationThe Discovery Foundation Grant (MNU), Grimes Foundation Endowment Award (MNU)
Characterizing the specificity of E3 ubiquitin ligase MARCH1 for CD98
Authors: Najera Susana Sarai , Ablack Jailal , Ginsberg Mark 
Published: November 2023

CD98 is a type II transmembrane protein that supports adhesive signaling and amino acid‐transport enabling rapid cell proliferation. Our genetic ablation studies have revealed that CD98 is essential for proliferation of multiple cell types including cancer cells. CD98 is found at the cell surface where it interacts with integrins and its light chains to amplify adhesive signaling and amino acid transport, respectively. Surface expression of CD98 is regulated post‐translationally by MARCH1 and MARCH8, members of a novel class of E3 ubiquitin‐ligases. MARCH1 limits CD98 dependent proliferation of lymphocytes, and potentially cancer cells, by selectively targeting CD98 for ubiquitination and subsequent lysosomal degradation. However, the mechanism by which MARCH1 targets and interacts with CD98 is still poorly understood. Our goal is to decipher how MARCH1 achieves specificity for its substrates using CD98 as a model substrate. Since MARCH1 isoforms have unique N‐termini, we initially hypothesized that the N‐terminus of MARCH1 mediated the interaction with CD98. Ectopic expression of either isoform of MARCH1 in HeLa cells resulted in down regulation of CD98 from the cell surface, while ectopic expression of a catalytically inactive mutant of MARCH1 had no effect. Thus, the N‐terminus alone does not determine MARCH1 specificity for CD98. We next expanded our hypothesis to include other domains of MARCH1 that could be responsible for imparting specificity for CD98. To test this hypothesis, we generated chimeras of MARCH1 and MARCH3 (MARCH3 has no specificity for CD98) to determine which regions of MARCH1 impart specificity for CD98. Our preliminary results confirmed that the N‐terminus of MARCH1 alone, is not sufficient for targeting of CD98. Likewise, the RING domain, the transmembrane domains and C‐terminus of MARCH1 on their own are not sufficient to impart CD98 specificity to MARCH3. Together these observations suggest that the domains of MARCH1 likely work in combination to selectively interact with CD98 and target it for degradation.Support or Funding InformationResearch reported in this poster was supported by the National Cancer Institute of the National Institutes of Health under award numbers: U54CA132384 & U54CA132379
The potential role of NMDA receptor regulating TGF‐β/Smad pathway in radiation‐induced resistance in glioblastoma multiforme
Authors: Liu Chan‐Chuan , Wu Sheng‐Nan , Sze Chun‐I 
Published: November 2023

Glioblastoma multiforme (GBM) is the most lethal primary brain cancer with poor prognosis despite the aggressive therapeutic treatment. Infiltrative growth, resistance to therapy, genetic heterogeneity, and the existence of cancer stem cells lead to tumor recurrence and poor outcome. The current standard of therapy is surgery followed by radiation and/or chemotherapy. Unfortunately, tumor cells establish resistance to the therapy. Glutamate is a neurotransmitter that induces post‐synaptic excitation of neurons for the cell communication. Glutamate activating ionotropic and metabotropic receptor is also essential in tissue development. Glutamate receptors are classified into NMDA, AMPA and kainate receptors. The activation of glutamate receptor involves in regulation of cell death and survival, proliferation, and migration. Previous studies revealed that activation of NMDA receptor via glutamate regulates proliferation and invasion in gliomas. In addition, cross talk between NMDA receptor and TGF‐β/Smad signaling leads to cell differentiation. We hypothesize that NMDA receptor may play role in the generation of radiation‐induced resistance in GBM.Our laboratory had been using growth inhibition 50% and consecutive irradiation to establish radiation resistant GBM cell lines. Parts of radiation resistant cells showed morphological changes and we defined these cells as transformed cells. Immunocytochemistry staining showed the different expression patterns of NMDA receptor subunits, and its adaptor protein PSD95 between transformed cells and non‐transformed cells. In Western blotting results, we found that the expression of NMDA receptor and PSD95 are decreased in the development of radiation resistance. We also found that NMDA receptor related signaling proteins, such as nNOS, Akt, and ERK, are also decreased. In our previous discovery, radiation activates TGF‐β/Smad signaling which leads to mesenchymal differentiation. On the other hand, inhibition of TGF‐β/Smad pathway reverses mesenchymal differentiation in radiation resistant cell lines. In our current studies, TGF‐β treatment results in parental GBM cell lines reduced expression of the NMDA receptor and PSD95, however, the inhibition of TGF‐β receptor have no influence on these changes. We conclude that the decrement of NMDA receptor and its related pathways help GBM cells establish radiation resistance which suggests that NMDA receptor may regulate TGF‐β/Smad signaling.
An Ageless Wonder: Decreasing Cardiac CapZ Prevents Myocardial Decline in Aged Mice
Authors: Pyle W. Glen , Lorenzen‐Schmidt Ilka , Reitz Cristine J , Alibhai Faisal J , Martino Tami A. 
Published: November 2023

Ageing populations are a worldwide trend: by 2030 the number of people over 65 years of age will double and this cohort will continue to increase by another 50% by 2050. Age is the primary risk factor for cardiovascular disease with a 22% increase in risk every 5 years after age 65. Mitigating the functional decline of the heart that comes with advanced age will reduce susceptibility to heart failure and prolong lifespans. The actin capping protein ‘CapZ’ is a key regulator of molecular remodeling in ageing skeletal muscle. Whether it plays a similar role in the heart has never been investigated. Our objective was to determine if cardiac CapZ is altered in the hearts of aged mice, and to ascertain the impact of these changes. Wildtype C57Bl6 mice of both sexes were aged to 21 months and cardiac function determined by echocardiography. Left ventricular ejection fraction decreased from 77.2 ± 0.3% to 69.7 ± 0.8% (p<0.05) in male mice at 4 and 21 months of age respectively, whereas female mice showed no significant changes with ageing. Left ventricular fractional shortening showed a similar reduction in aged male mice, declining from 40.2 ± 0.3% to 34.1 ± 0.6% (p<0.05) in male mice aged 4 and 21 months. Again, females maintained fractional shortening with ageing. Immunoblot analysis revealed no differences in cardiac CapZ expression in male myocardial samples between 4 and 21 month old mice. Samples from female hearts, however, showed a significant reduction in CapZ protein levels. To determine if the natural reduction in CapZ protein levels seen in female hearts mitigated the functional decline of ageing, we measured myocardial function of CapZ‐deficient transgenic mice by echocardiography. While female CapZ‐deficient transgenic mice again exhibited no differences in cardiac performance between 4 and 21 month old animals, the functional decline that characterized aged male wildtype mice was significantly alleviated in CapZ‐deficient transgenic mice. It has been proposed that one factor contributing to the age‐dependent decline in cardiac performance is a reduction in beta‐adrenergic reserve. We have previously shown that CapZ influences beta‐adrenergic signaling in the heart. We measured the myocardial response to beta‐adrenergic stimulation of Langendorff‐perfused murine hearts and discovered that aged CapZ‐deficient transgenic mice retain a robust positive inotropic response to myocardial beta‐adrenergic activation, unlike aged wildtype mice. These studies represent the first report showing that decreased cardiac CapZ is an endogenous mechanism by which the hearts of female mice protect themselves against ageing, and demonstrate that reduced cardiac CapZ is sufficient to mitigate the functional decline of advanced age.Support or Funding InformationCIHR Catalyst Grant
The role of the sphingolipid metabolism pathway in healthy aging
Authors: Hrobuchak Hannah P , Ardasheva Anastasia P , Servello Dustin P , Nolan Abby P , Chan Jason P 
Published: November 2023

Animals are exposed to stressors, such as temperature and chemical stress, which may contribute to their aging. Organisms respond to stress by mobilizing cell‐signaling pathways, which are critical to control cell function and stress response. One specific type of pathway is the sphingolipid‐signaling pathway, which interconverts between different sphingolipids that can function as critical cell survival and cell death factors. Two critical enzymes are sphingosine kinase and ceramide kinase, which converts a cell death signals to cell survival factors (ceramide‐1‐phosphate and sphingosine‐1‐phosphate). However, less is known regarding the role of these lipid signaling pathways in healthspan, or healthy aging in animals. To examine how these enzymes mediate healthspan, we use the genetic model organism C. elegans to examine sphingolipid metabolism. We identified that sphingosine kinase, in particular, is important for mediating lifespan and healthspan. Through neuromuscular and heat stress assays, we determined that locomotor behavior and thermoregulatory responses of animals declined at greater rates when they lacked sphingosine kinase. However, we found that there is no differences in locomotor response in ceramide kinase mutants. Further examination of how sphingosine kinase and ceramide kinase loss at developmental and post‐reproductive windows will help us better understand the role of lipids in aging.Support or Funding InformationNational Institute On Aging of the National Institutes of Health, Award Number R15AG052933
Immunohistofluorescence Localization of Biogenic Amine Receptors in Ganglia and Tissues of the Bivalve Mollusc, Mytilus edulis
Authors: Cummings Delilah , Jacobs Maxine , Carroll Margaret A , Catapane Edward J 
Published: November 2023

Biogenic amines serve as neurotransmitters in both sensory and motor systems in a variety of invertebrates. In bivalves, particularly Mytilus edulis, the physiological roles of serotonin and dopamine have been well studied especially with respect to gill and heart. Recently, in the related bivalve Crassostrea virginica, other biogenic amines, including GABA and histamine along with their receptors, were demonstrated to be present, and these amines are being shown to have physiological roles. In light of this we hypothesize that M. edulis would have receptors for the biogenic amines, serotonin, dopamine, histamine and GABA. We tested this using immunohistofluorescence techniques. Primary antibodies to serotonin, dopamine, histamine and GABA receptors, and secondary FITC‐labeled antibodies were purchased from Abcam and Santa Cruz Biotechnology, and used to visualize the presence or absence of these biogenic amine receptors in cerebral ganglia, visceral ganglia, and the animals' innervated gill and mantle. Briefly, tissues were dissected, snap frozen, sectioned on a cryostat, fixed with EDAC (N‐Ethyl‐N′‐(3‐dimethylaminopropyl) carbodiimide hydrochloride), treated with blockers, and incubated with primary, then secondary antibodies. Controls were treated the same way except without primary antibodies. Treated cryostat sections were viewed with a Zeiss epilume fluorescence microscope fitted with a ProgRes C3 Peltier cooled camera, or with a Leica epilume fluorescence microscope fitted with a Leica DFC400 camera. Both microscopes had 100 watt mercury lamps and FITC excitation/emission filters (Ex 470 nm, Em 525 nm). The results show serotonin 2A‐like, dopamine D2‐like, histamine H2‐like and GABA R1‐6‐like receptors present in cerebral ganglia, visceral ganglia, gill and in the sensory tentacles of the mantle rim. This comparative study provides new knowledge of the neurobiology of M. edulis by confirming the presence of these biogenic amine receptors in the animals' nervous system and innervated organs, in a comparable manner to what has been reported in C. virginica. It also provides a foundation to further study the physiological roles for histamine and GABA in M. edulis.Support or Funding InformationThis work was supported in part by grant 2R25GM06003 of the Bridge Program of NIGMS, grant 690340047 of PSC‐CUNY and a Carnegie Foundation award.
High‐throughput Functional Annotation of the Caenorhabditis elegans Neural Network
Authors: Aoki Wataru , Yokoyama Haruki , Matsukura Hidenori , Ueda Mitsuyoshi 
Published: November 2023

In order to investigate the relationship between behavior and neural networks, such a technology that can artificially control the activity of the particular neuron is required. Once we artificially manipulate the activity of neurons, we will be able to examine in detail how the neural networks function by evaluating how the artificial manipulation influences the behavior.Optogenetics is a technology that is able to freely control the activity of neurons with opsin by light irradiation. Optogenetics provides a methodology that can precisely analyze the relationship between behavior and neural network, and has become a trigger of rapid progress of brain research in the circuit‐centric approaches.However, there are some weaknesses in the optogenetics.1. Optogenetics requires a hypothesis, because we have to consider in advance which group of neurons is related to particular behavior when choosing promoter regulating opsin expression. For this reason, although it is a powerful method for validation and development of existing hypothesis, a completely new discovery is hard to be achieved. 2. Throughput is low. We need to create different transgenic animals depending on the hypothesis to be tested. 3. Single cell level analysis is difficult. Cell‐specific promoter does not usually exist. In addition, it is difficult to converge light on the single neuron of moving animals.In order to overcome these weaknesses, we propose a new method that does not need any hypothesis, can be high throughput and can analyze in single cell resolution (Aoki et al. submitted). A key technology for this methodology is Cre/lox system. By applying Cre/lox system, we have established a methodology to randomize single neuron that expresses opsin, among the all neurons that construct the neural network.In this methodology, we conduct behavior experiment after opsin expressions are randomized, and identify the neuron that influences the behavior “later”. Therefore, it does not require the hypothesis in advance. In addition, since we can create a large number of individuals that have randomized opsin expression pattern, from single individual, our method is feasible for high throughput analysis. Since expression of opsin is determined in each cell at random, single cell level analysis is feasible.Here we present stochastic expression of opsin and high‐throughput annotation of phenotype using C. elegans as an experimental model.
Attenuation of Glia Maturation Factor Enhances Mitochondrial Biogenesis, Decreases Oxidative Stress and Apoptotic Cell Death in N27 Dopaminergic Neurons
Authors: Selvakumar Govindhasamy Pushpavathi , Kempuraj Duraisamy , Thangavel Ramasamy , Ahmed Mohammad Ejaz , Zaheer Smita , Iyer Shankar S , Zaheer Asgar 
Published: November 2023

Parkinson's disease (PD) is a progressive loss of dopamine producing neurons in the brain, that leads to motor complications. But the exact molecular events underlying the neuronal death are still not known. Glia maturation factor (GMF), a brain specific protein, plays a crucial role in PD progression. Mitochondrial abnormalities and oxidative stress triggers apoptotic dopaminergic degeneration in PD. Peroxisome proliferator activated receptor‐gamma coactivator‐(PPARGC‐1α or PGC‐1 α) acts as a key regulating factor for the mitochondrial biogenesis and respiration in dopaminergic cells in PD. We examined if attenuation of GMF expression enhances the expression of PGC‐1 α and reduces oxidative stress, mitochondrial dysfunction and apoptotic cell death in dopaminergic neurons. N27 dopaminergic neuronal cells were treated with GMF (100 ng/ml) for 24 h and analyzed for PGC‐1α expression by immunocytochemistry. Cell lysates were also prepared from these cells to estimate oxidative stress markers (glutathione and superoxide dismutase) using microplate reader, and expression of apoptotic (Bax cytochrome‐c, caspases 3, 7, 8,9) and anti‐apoptotic (Bcl‐2) markers by immunoblotting. GMF treatment for 24 h reduces the expression of PGC‐1α and mitochondrial complexes in N27 cells as compared with control cells. GMF also increased oxidative stress and depolarized mitochondrial membrane potential (MMP) in these cells. Furthermore, GMF increases Bax and reduces Bcl‐2 expression, stimulates the release of cytochrome‐c leading to activation of initiator procaspase‐9 and effector procaspases in N27 cells. These results suggest a crucial role of GMF in mitochondrial abnormalities and oxidative stress‐mediated apoptosis through PGC‐1α in dopaminergic neurons. We conclude that GMF is a critical factor for PD pathology and may be a potential therapeutic target to inhibit PD progression.Support or Funding InformationThis work was supported by Veteran Affairs Merit Award I01BX002477 and National Institutes of Health Grants AG048205 and NS073670