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Item Effect of Epigallocatechin-3-gallate on Skeletal and Cognitive Phenotypes in a Down Syndrome Mouse Model(2014) Abeysekera, Irushi Shamalka; Roper, Randall J.; Goodlett, Charley; Li, Jiliang; Yost, RobertDown syndrome (DS), a genetic disorder that affects ~1 in 700 live births, is caused by trisomy of human chromosome 21 (Hsa21). Individuals with DS are affected by a wide spectrum of phenotypes which vary in severity and penetrance. However, cognitive and skeletal impairments can be commonly observed in all individuals with DS. To study these phenotypes, we utilized the Ts65Dn mouse model that carries three copies of approximately half the gene orthologs found on Hsa21 and exhibit similar phenotypes as observed in humans with DS. Individuals with DS and Ts65Dn mice have deficits in bone mineral density (BMD), bone architecture, bone strength, learning and memory. Over-expression of DYRK1A, a serine-threonine kinase encoded on Hsa21, has been linked to deficiencies in DS bone homeostasis and cognition. Epigallocatechin-3-gallate (EGCG), an aromatic polyphenol found in high concentrations in green tea, is a selective inhibitor of DYRK1A activity. Normalization of DYRK1A activity by EGCG therefore may have the potential to ameliorate skeletal and cognitive deficits. We hypothesized that supplements containing EGCG obtained from health food stores/ online vendors will not be as effective as EGCG from a chemical company in correcting bone deficits associated with DS. Our results suggest that EGCG improves the bone mineral density of trisomic femurs significantly better than the supplements while the EGCgNOW supplement from NOW FOODS improves trabecular and cortical bone structure. The results from HPLC analysis of supplements showed the presence of other catechins in EGCgNOW and degradation analysis revealed the rapid degradation of supplements. Therefore we hypothesize that the presence of EGCG degradation products and other green tea catechins in supplements may play a role in the differential skeletal effects we observed. We further hypothesized that a three week treatment of adolescent mice with EGCG will lead to an improvement in the learning and memory deficits that are observed in trisomic animals in comparison to control mice. However, our results indicate that three weeks of low-dose EGCG treatment during adolescence is insufficient to improve hippocampal dependent learning and memory deficits of Ts65Dn mice. The possibility remains that a higher dose of EGCG that begins at three weeks but lasts throughout the behavioral test period may result in improvement in learning and memory deficit of Ts65Dn mice.Item Nitrous oxide emission from riparian buffers in agricultural landscapes of Indiana(2014-02-25) Fisher, Katelin Rose; Babbar-Sebens, Meghna; Jacinthe, Pierre-André; Vidon, Philippe G.Riparian buffers have well documented capacity to remove nitrate (NO3-) from runoff and subsurface flow paths, but information on field-scale N2O emission from these buffers is lacking. This study monitored N2O fluxes at two agricultural riparian buffers in the White River watershed (Indiana) from December 2009 to May 2011 to assess the impact of landscape and hydrogeomorphologic factors on emission. Soil chemical and biochemical properties were measured and environmental variables (soil temperature and moisture) were monitored in an attempt to identify key drivers of N2O emission. The study sites included a mature riparian forest (WR) and a riparian grass buffer (LWD); adjacent corn fields were also monitored for land-use comparison. With the exception of net N mineralization, most soil properties (particle size, bulk density, pH, denitrification potential, organic carbon, C:N) showed little correlation with N2O emission. Analysis of variance (ANOVA) identified season, land-use (riparian buffer vs. crop field), and site geomorphology as major drivers of N2O emission. At both study sites, N2O emission showed strong seasonal variability; the largest emission peaks in the riparian buffers (up to 1,300 % increase) and crop fields (up to 3,500 % increase) occurred in late spring/early summer as a result of flooding, elevated soil moisture and N-fertilization. Nitrous oxide emission was found to be significantly higher in crop fields than in riparian buffers at both LWD (mean: 1.72 and 0.18 mg N2O-N m-2 d-1) and WR (mean: 0.72 and 1.26 mg N2O-N m-2 d-1, respectively). Significant difference (p=0.02) in N2O emission between the riparian buffers was detected, and this effect was attributed to site geomorphology and the greater potential for flooding at the WR site (no flooding occurred at LWD). More than previously expected, the study results demonstrate that N2O emission in riparian buffers is largely driven by landscape geomorphology and land-stream connection (flood potential).Item The role of STAT3 in osteoclast mediated bone resorption(2014-08-01) Himes, Evan; Li, Jiliang; Yost, Robert; Kacena, Melissa A.; Atkinson, SimonSignal Transducer and Activator of Transcription 3 (STAT3) is known to be related to bone metabolism. Mutation of STAT3 causes a rare disorder in which serum levels of IgE are elevated. This causes various skeletal problems similar to osteoporosis. To examine the effect of STAT3 in the osteoclast, we obtained two osteoclast specific STAT3 knockout mouse models: one using the CTSK promoter to drive Cre recombinase and another using a TRAP promoter. Examination of these mice at 8 weeks of age revealed a decreased trabecular bone volume in CTSK specific STAT3 knockout mice along with a slight decrease in osteoclast number in both CTSK and TRAP specific STAT3 knockout females. We also noticed changes in bone mineral density and bone mechanical strength in females. These data suggest that STAT3 plays a part in the function of the osteoclast.Item Self-assembling peptide-laden electrospun scaffolds for guided mineralized tissue regeneration(Elsevier, 2022) de Souza Araújo, Isaac J.; Ferreira, Jessica A.; Daghrery, Arwa; Ribeiro, Juliana S.; Castilho, Miguel; Puppin-Rontani, Regina M.; Bottino, Marco C.; Biomedical and Applied Sciences, School of DentistryObjectives: Electrospun scaffolds are a versatile biomaterial platform to mimic fibrillar structure of native tissues extracellular matrix, and facilitate the incorporation of biomolecules for regenerative therapies. Self-assembling peptide P11-4 has emerged as a promising strategy to induce mineralization; however, P11-4 application has been mostly addressed for early caries lesions repair on dental enamel. Here, to investigate P11-4's efficacy on bone regeneration, polymeric electrospun scaffolds were developed, and then distinct concentrations of P11-4 were physically adsorbed on the scaffolds. Methods: P11-4-laden and pristine (P11-4-free) electrospun scaffolds were immersed in simulated body fluid and mineral precipitation identified by SEM. Functional groups and crystalline phases were analyzed by FTIR and XRD, respectively. Cytocompatibility, mineralization, and gene expression assays were conducted using stem cells from human exfoliated deciduous teeth. To investigate P11-4-laden scaffolds potential to induce in vivo mineralization, an established rat calvaria critical-size defect model was used. Results: We successfully synthesized nanofibrous (∼ 500 nm fiber diameter) scaffolds and observed that functionalization with P11-4 did not affect the fibers' diameter. SEM images indicated mineral precipitation, while FTIR and XRD confirmed apatite-like formation and crystallization for P11-4-laden scaffolds. In addition, P11-4-laden scaffolds were cytocompatible, highly stimulated cell-mediated mineral deposition, and upregulated the expression of mineralization-related genes compared to pristine scaffolds. P11-4-laden scaffolds led to enhanced in vivo bone regeneration after 8 weeks compared to pristine PCL. Significance: Electrospun scaffolds functionalized with P11-4 are a promising strategy for inducing mineralized tissues regeneration in the craniomaxillofacial complex.