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Item Changes in immunofluorescence staining during islet regeneration in a cystic fibrosis-related diabetes (CFRD) ferret model(Taylor & Francis, 2024) Mohammed, Sawash M.; Bone, Robert N.; Aquino, Jacqueline Del Carmen; Mirmira, Raghavendra G.; Evans-Molina, Carmella; Ismail, Heba M.; Anatomy, Cell Biology and Physiology, School of MedicineBackground: Knockout (KO) ferrets with the cystic fibrosis transmembrane conductance regulator (CFTR) exhibit distinct phases of dysglycemia and pancreatic remodeling prior to cystic fibrosis-related diabetes (CFRD) development. Following normoglycemia during the first month of life (Phase l), hyperglycemia occurs during the subsequent 2 months (Phase Il) with decreased islet mass, followed by a period of near normoglycemia (Phase Ill) in which the islets regenerate. We aimed to characterize islet hormone expression patterns across these Phases. Methods: Immunofluorescence staining per islet area was performed to characterize islet hormone expression patterns in age matched CFTR KO and wild type (WT) ferrets, focusing on the first three phases. Results: In Phase I, insulin staining intensity was higher in CF (p < 0.01) than WT but decreased in Phase III (p < 0.0001). Glucagon was lower in CF during Phases I and increased in Phase III, while proinsulin decreased (p < 0.0001) Phases II and III. CF sections showed lower proinsulin-to-insulin ratio in Phase I (p < 0.01) and in Phase III (p < 0.05) compared to WT. Conversely, glucagon-to-insulin ratio was lower in CF in Phase I (p < 0.0001) but increased in Phase III (p < 0.0001). Mender's coefficient overlap showed higher overlap of insulin over proinsulin in CF sections in Phase II (p < 0.001) and Phase III (p < 0.0001) compared to WT. Mender's coefficient rate was higher in CF sections during Phase II (p < 0.001). Conclusion: CF ferret islets revealed significant immunofluorescent staining changes compared to WT during various phases of disease, providing insights into CRFD pathophysiology.Item Subcutaneous Nerve Stimulation Reduces Sympathetic Nerve Activity in Ambulatory Dogs with Myocardial Infarction(Elsevier, 2020-07) Yuan, Yuan; Zhao, Ye; Wong, Johnson; Tsai, Wei-Chung; Jiang, Zhaolei; Kabir, Ryan A.; Han, Seongwook; Shen, Changyu; Fishbein, Michael C.; Chen, Lan S.; Chen, Zhenhui; Everett, Thomas H., IV.; Chen, Peng-Sheng; Medicine, School of MedicineBackground: Subcutaneous nerve stimulation (ScNS) remodels the stellate ganglion and reduces stellate ganglion nerve activity (SGNA) in dogs. Acute myocardial infarction (MI) increases SGNA through nerve sprouting. Objective: The purpose of this study was to test the hypothesis that ScNS remodels the stellate ganglion and reduces SGNA in ambulatory dogs with acute MI. Methods: In the experimental group, a radio transmitter was implanted during the first sterile surgery to record nerve activity and an electrocardiogram, followed by a second sterile surgery to create MI. Dogs then underwent ScNS for 2 months. The average SGNA (aSGNA) was compared with that in a historical control group (n = 9), with acute MI monitored for 2 months without ScNS. Results: In the experimental group, the baseline aSGNA and heart rate were 4.08±0.35 μV and 98±12 beats/min, respectively. They increased within 1 week after MI to 6.91±1.91 μV (P=.007) and 107±10 beats/min (P=.028), respectively. ScNS reduced aSGNA to 3.46±0.44 μV (P<.039) and 2.14±0.50 μV (P<.001) at 4 and 8 weeks, respectively, after MI. In comparison, aSGNA at 4 and 8 weeks in dogs with MI but no ScNS was 8.26±6.31 μV (P=.005) and 10.82±7.86 μV (P=0002), respectively. Immunostaining showed confluent areas of remodeling in bilateral stellate ganglia and a high percentage of tyrosine hydroxylase-negative ganglion cells. Terminal deoxynucleotidyl transferase dUTP nick end labeling was positive in 26.61%±11.54% of ganglion cells in the left stellate ganglion and 15.94%±3.62% of ganglion cells in the right stellate ganglion. Conclusion: ScNS remodels the stellate ganglion, reduces SGNA, and suppresses cardiac nerve sprouting after acute MI.Item Using TMAs (Tissue MicroArrays) to Evaluate GLS, GLUL, and CAV 1 Immunostaining in Breast Cancer(Office of the Vice Chancellor for Research, 2016-04-08) Finnearty, Courtney; Shajahan-Haq, Ayesha; Nakshatri, Hari; Sandusky, George E.Approximately 1 out of 8 women in the United States will develop invasive breast cancer over the course of their lifetime. Breast cancer has a greater potential of being cured if diagnosed in the earlier phases. We evaluated three well-recognized biomarkers, GLUL, GLS, and Cav 1 (glutamine synthetase, glutaminase, caveolin-1) in 14 TMA (tissue Microarrays). The tissues were normal breast and various subtypes of breast carcinoma by immunohistochemistry (IHC) to determine expression and localization in cancerous tissues in breast carcinoma cases. Approximately 80 to 90 breast biopsies in each of the 14 breast TMA immunostaining were evaluated with the GLUL, GLS, and Cav 1 antibodies. With GLS, immunostaining was seen in most tumor cells (mainly cytoplasm and nucleus) and the stain was clean with no background except in cases that had lymphocytes in the core along with the tumor cells. With GLUL, immunostaining was seen in most tumor cells (mainly cytoplasm and nucleus) and the stain was clean with no background except in cases that had lymphocytes in the core along with the tumor cells. Cav1 was seen only in the endothelial cells in blood vessel walls and some smooth muscle cells in small arterioles in the stroma and surrounding normal ducts, DCIS, and some invasive carcinoma tumor clusters. This information from the immunostains was obtained after analyzing 14 tissue microarrays which is not only time effective but cost effective when analyzing multiple research cases from cancer patients. The data for the three antibodies are currently being analyzed by the biostatistics core group and correlated with the severity of the breast cancer disease with multiple patient demographics.