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Item ADF/Cofilin Activation Regulates Actin Polymerization and Tension Development in Canine Tracheal Smooth Muscle(2009-09-03T15:28:09Z) Zhao, Rong; Gunst, Susan J.; Atkinson, Simon J.; Elmendorf, Jeffrey S.; Sturek, Michael S.The contractile activation of airway smooth muscle tissues stimulates actin polymerization and the inhibition of actin polymerization inhibits tension development. Actin depolymerizing factor (ADF) and cofilin are members of a family of actin–binding proteins that mediate the severing of F–actin when activated by dephosphorylation at serine 3. The role of ADF/cofilin activation in the regulation of actin dynamics and tension development during the contractile activation of airway smooth was evaluated in intact canine tracheal smooth muscle tissues. Two–dimensional gel electrophoresis revealed that ADF and cofilin exist in similar proportions in the muscle tissues and that approximately 40% of the total ADF/cofilin in unstimulated tissues is phosphorylated (inactivated). Phospho–ADF/cofilin decreased concurrently with tension development in response to stimulation with acetylcholine (ACh) or potassium depolarization indicating the activation of ADF/cofilin. Expression of an inactive phospho–cofilin mimetic (cofilin S3E), but not WT cofilin in the smooth muscle tissues inhibited endogenous ADF/cofilin dephosphorylation and ACh–induced actin polymerization. Expression of cofilin S3E in the tissues depressed tension development in response to ACh, but it did not affect myosin light chain phosphorylation. The ACh–induced dephosphorylation of ADF/cofilin required the Ca2+–dependent activation of calcineurin (PP2B). Expression of Slingshot (SSH) inactive phosphatase (C393S) decreased force development and cofilin dephosphorylation. Activation of ADF/cofilin was also required for the relaxation of tracheal muscle tissues induced by forskolin and isoproterenol. Cofilin activation in response to forskolin was not Ca2+–dependent and was not inhibited by calcineurin inhibitors, suggesting it was regulated by a different mechanism. Cofilin activation is required for actin dynamics and tension development in response to the contractile stimulation of tracheal smooth muscle and is regulated by both contractile and relaxing stimuli. These concepts are critical to understanding the mechanisms of smooth muscle contraction and relaxation, which may provide novel targets for therapeutic intervention in the treatment of abnormal airway responsiveness.Item Effect of interim fixed prosthodontics materials and flowable composite resins on polymerization of polyvinyl siloxane impressions(2017) Alsayed, Hussain D.; Platt, Jeffrey A.; Levon, John A.; Haug, Steven P.; Brown, David T.Item Effect of lithium disilicate ceramic thickness, shade and translucency on transmitted irradiance and knoop microhardness of a light cured luting resin cement(Springer, 2021-07-31) Borges, Lincoln Pires Silva; Borges, Gilberto Antônio; Correr, Américo Bortolazzo; Platt, Jeffrey A.; Kina, Sidney; Correr-Sobrinho, Lourenço; Costa, Ana Rosa; Biomedical and Applied Sciences, School of DentistryThis in vitro study evaluates the influence of pressed lithium disilicate thickness, shade and translucency on the transmitted irradiance and the Knoop microhardness (KHN) of a light-cured resin cement at two depths. One hundred and thirty-five ceramic discs of IPS e.max Press (Ivoclar Vivadent) were fabricated and divided into twenty-seven groups (n = 5) according to the association between translucency: HT (hight translucency), LT (low translucency), and MO (medium opacity); shade: BL2, A1 and A3.5; and thickness: 0.5 mm, 1.5 mm, and 2.0 mm. One side of each ceramic disc was finished, polished and glazed. The irradiance (mW/cm²) of a multiwave LED light curing unit (Valo, Ultradent) was evaluated with a potentiometer (Ophir 10ª-V2-SH, Ophir Optronics) without (control group) or with interposition of ceramic samples. The microhardness of Variolink Esthetic LC resin cement (Ivoclar Vivadent) was evaluated after 24 h at two depths (100 μm and 700 μm). Data were submitted to ANOVA followed by Tukey's test (α = 0.05). Irradiance and KHN were significantly influenced by ceramic thickness (p < 0.0001), shade (p < 0.001), translucency (p < 0.0001) and depth (p < 0.0001). Conclusions: the interposition of increasing ceramic thicknesses significantly reduced the irradiance and microhardness of resin cement. Increased depth in the resin cement showed significantly reduced microhardness for all studied groups. Increased ceramic opacity reduced the KHN of the resin cement at both depths for all ceramic thicknesses and shades.Item The effect of polymerization methods and fiber types on the mechanical behavior of fiber-reinforced composite resin(2015) Huang, Nan-Chieh; Chu, Tien-Min Gabriel; Hara, Anderson T.; Brown, David T.; Bottino, Marco C.; Levon, John A.Background: Interim restoration for a lost anterior tooth is often needed for temporary esthetic and functional purposes. Materials for interim restorations usually have less strength than ceramic or gold and can suffer from fracture. Several approaches have been proposed to reinforce interim restorations, among which fiber reinforcement has been regarded as one of the most effective methods. However, some studies have found that the limitation of this method is the poor polymerization between the fibers and the composite resin, which can cause debonding and failure. 64 Purpose: The purpose of this study was to investigate the effects of different polymerization methods as well as fiber types on the mechanical behavior of fiberreinforced composite resin. Material and Methods: A 0.2-mm thick fiber layer from strip fibers or mesh fibers embedded in uncured monomers w as fabricated with polymerization (two-step method) or without polymerization (one-step method), on top of which a 1.8-mm composite layer was added to make a bar-shape sample, followed by a final polymerization. Seventy-five specimens were fabricated and divided into one control group and four experimental groups (n=15), according to the type of glass fiber (strip or mesh) and polymerization methods (one-step or two-step). Specimens were tested for flexural strength, flexural modulus, and microhardness. The failure modes of specimens were observed by scanning electron microscopy (SEM). Results: The fiber types showed significant effect on the flexural strength of test specimens (F = 469.48; p < 0.05), but the polymerization methods had no significant effect (F = 0.05; p = 0.82). The interaction between these two variables was not significant (F = 1.73; p = 0.19). In addition, both fiber types and polymerization steps affected the flexural modulus of test specimens (F = 9.71; p < 0.05 for fiber type, and F = 12.17; p < 0.05 for polymerization method). However, the interaction between these two variables was not significant (F = 0.40; p = 0.53). Both fiber types and polymerization steps affected the Knoop hardness number of test specimens (F = 5.73; p < 0.05 for polymerization method. and F = 349.99; p < 0.05 for fiber type) and the interaction between these two variables was also significant (F = 5.73; p < 0.05). SEM images revealed the failure mode tended to become repairable while fiber reinforcement was 65 existed. However, different polymerization methods did not change the failure mode. Conclusion: The strip fibers showed better mechanical behavior than mesh fibers and were suggested for use in composite resin reinforcement. However, different polymerization methods did not have significant effect on the strength and the failure mode of fiber-reinforced compositeItem Effect of post-irradiation polymerization on selected mechanical properties of six direct resins(J-STAGE, 2022) Sochacki, Sabrina; Karczewski, Ashley; Platt, Jeffrey; Roberts, Howard W.; Biomedical Sciences and Comprehensive Care, School of DentistryThis study evaluated the post-irradiation mechanical property development of six resin composite-based restorative materials from the same manufacturer starting at 1 h post irradiation, followed by 24 h, 1 week, and 1 month after fabrication. Samples were stored in 0.2M phosphate buffered saline until testing. Flexural strength, flexural modulus, flexural toughness, modulus of resiliency, fracture toughness, and surface microhardness were performed at each time interval. Mean data was analyzed by Kruskal Wallis and Dunn's post hoc testing at a 95% level of confidence (α=0.05). Results were material specific but overall, all resin composite material mechanical properties were found to be immature at 1 h after polymerization as compared to that observed at 24 h. It may be prudent that clinicians advise patients, especially those receiving complex posterior composite restorations, to guard against overly stressing these restorations during the first 24 h.Item Effect of Rapid High-Intensity Light-Curing on Increasing Transdentinal Temperature and Cell Viability: An In Vitro Study(MDPI, 2024-05-22) Miranda, Samille Biasi; Lins, Rodrigo Barros Esteves; Santi, Marina Rodrigues; Denucci, Giovanna Corrêa; Silva, Cleyton Cézar Souto; da Silva, Silvana de Fátima Ferreira; Marques, Daniela de Araújo Viana; Montes, Marcos Antônio Japiassú Resende; Comprehensive Care and Allied Professions, School of DentistryBackground: This study investigated effects of rapid high-intensity light-curing (3 s) on increasing transdentinal temperature and cell viability. Methods: A total of 40 dentin discs (0.5 mm) obtained from human molars were prepared, included in artificial pulp chambers (4.5 × 5 mm), and subjected to four light-curing protocols (n = 5), with a Valo Grand light curing unit: (i) 10 s protocol with a moderate intensity of 1000 mW/cm2 (Valo-10 s); (ii) 3 s protocol with a high intensity of 3200 mW/cm2 (Valo-3 s); (iii) adhesive system + Filtek Bulk-Fill Flow bulk-fill composite resin in 10 s (FBF-10 s); (iv) adhesive system + Tetric PowerFlow bulk-fill composite resin in 3 s (TPF-3 s). Transdentinal temperature changes were recorded with a type K thermocouple. Cell viability was assessed using the MTT assay. Data were analyzed using one-way ANOVA and Tukey tests for comparison between experimental groups (p < 0.05). Results: The 3 s high-intensity light-curing protocol generated a higher temperature than the 10 s moderate-intensity standard (p < 0.001). The Valo-10 s and Valo-3 s groups demonstrated greater cell viability than the FBF-10s and TPF-3 s groups and statistical differences were observed between the Valo-3 s and FBF-10 s groups (p = 0.023) and Valo-3 s and TPF-3 s (p = 0.025), with a potential cytotoxic effect for the FBF-10 s and TPF-3 s groups. Conclusions: The 3 s rapid high-intensity light-curing protocol of bulk-fill composite resins caused a temperature increase greater than 10 s and showed cell viability similar to and comparable to the standard protocol.Item In vitro evaluation of polymerization energy for bulk fill composites(2016-05) AlRasheed, Rawan S.; Platt, Jeffery A.; Wallace, Joseph; Matis, Bruce A.; Cook, Norman Blaine; Chu, Tien-Min GabrielRecently, the concept of “bulk-fill” resin-based composites (RBCs) has been re-emphasized, with claimed improvements in depth of cure (DOC) with similar mechanical properties and comparable adaptation to walls and margins relative to conventional composite. More research is needed to carefully examine the properties of these new materials. The objective of this study was to measure the light energy, microhardness (VHN), and elastic modulus across the depth of one conventional and three bulk-fill RBCs. Materials and Methods: Three commercially available bulk-fill RBCs (Tetric EvoCeram Bulk Fill [TE], SonicFill [SF], X-tra fill[XF]) and one conventional RBC (Premise [PR]) were evaluated (n = 10). DOC (using Vickers’s microhardness), elastic modulus (using atomic force microscopy), and the mean irradiance and total light energy transmitted through different thicknesses of RBC were measured by a spectrometer. The effects of group, location, and curing depth on VHN were analyzed using mixed-model ANOVA. Elastic modulus and light energy comparisons were made using two-way ANOVA, with a significance level of 5 percent. Results: There was a significant difference in the depths for the mean irradiance and total energy between different depths in all materials. All materials achieved the manufacturers’ claimed DOC. XF had the highest DOC with 7 mm and a light energy of 0.56± 0.02 J/cm2 at 7 mm. PR had the lowest DOC with 3 mm and a light energy of 0.84 ±0.12 J/cm2 at 3 mm. The elastic modulus showed significant variation in depth profiles that were different than the DOC. Significance: The manufacturers’ claims for bulk-fill DOC were achieved using a microhardness method. However, this method failed to detect the quality of the polymerization. Assessment of the elastic modulus using AFM is a promising method for greater understanding of the polymerization.Item Involvement of Collapsin Response Mediator Protein 2 in Posttraumatic Sprouting in Acquired Epilepsy(2014) Wilson, Sarah Marie; Oxford, Gerry S.; Khanna, Rajesh; Jen, Joanna; Xu, Zao C.; Jin, Xiao-MingPosttraumatic epilepsy, the development of temporal lobe epilepsy (TLE) following traumatic brain injury, accounts for 20% of symptomatic epilepsy. Reorganization of mossy fibers within the hippocampus is a common pathological finding of TLE. Normal mossy fibers project into the CA3 region of the hippocampus where they form synapses with pyramidal cells. During TLE, mossy fibers are observed to innervate the inner molecular layer where they synapse onto the dendrites of other dentate granule cells, leading to the formation of recurrent excitatory circuits. To date, the molecular mechanisms contributing to mossy fiber sprouting are relatively unknown. Recent focus has centered on the involvement of tropomycin-related kinase receptor B (TrkB), which culminates in glycogen synthase kinase 3β (GSK3β) inactivation. As the neurite outgrowth promoting collapsin response mediator protein 2 (CRMP2) is rendered inactive by GSK3β phosphorylation, events leading to inactivation of GSK3β should therefore increase CRMP2 activity. To determine the involvement of CRMP2 in mossy fiber sprouting, I developed a novel tool ((S)-LCM) for selectively targeting the ability of CRMP2 to enhance tubulin polymerization. Using (S)-LCM, it was demonstrated that increased neurite outgrowth following GSK3β inactivation is CRMP2 dependent. Importantly, TBI led to a decrease in GSK3β-phosphorylated CRMP2 within 24 hours which was secondary to the inactivation of GSK3β. The loss of GSK3β-phosphorylated CRMP2 was maintained even at 4 weeks post-injury, despite the transience of GSK3β-inactivation. Based on previous work, it was hypothesized that activity-dependent mechanisms may be responsible for the sustained loss of CRMP2 phosphorylation. Activity-dependent regulation of GSK3β-phosphorylated CRMP2 levels was observed that was attributed to a loss of priming by cyclin dependent kinase 5 (CDK5), which is required for subsequent phosphorylation by GSK3β. It was confirmed that the loss of GSK3β-phosphorylated CRMP2 at 4 weeks post-injury was likely due to decreased phosphorylation by CDK5. As TBI resulted in a sustained increase in CRMP2 activity, I attempted to prevent mossy fiber sprouting by targeting CRMP2 in vivo following TBI. While (S)-LCM treatment dramatically reduced mossy fiber sprouting following TBI, it did not differ significantly from vehicle-treated animals. Therefore, the necessity of CRMP2 in mossy fiber sprouting following TBI remains unknown.Item LAYER BY LAYER NANOASSEMBLY OF COPPER INDIUM GALLIUM SELENIUM (CIGS) NANOPARTICLES FOR SOLAR CELL APPLICATION(2011-12) Hemati, Azadeh; El-Mounayri, Hazim; Agarwal, Mangilal; Varahramyan, KodyIn this research thesis, copper indium gallium selenium (CIGS) nanoparticles were synthesized from metal chlorides, functionalized to disperse in water, and further used in layer by layer (LbL) nanoassembly of CIGS films. CIGS nanoparticles were synthesized through the colloidal precipitation in an organic solvent. The peak and average sizes of the synthesized particles were measured to be 68 nm and 75 nm in chloroform, and 30 nm and 115 nm in water, respectively. Two methods were used to disperse the particle in water. In the first method the stabilizing agent oleylamine (OLA) was removed through multiple cleaning processes, and in the second method ligand exchange was performed with polystyrene sulfonate (PSS). Zeta potential of CIGS nanoparticles dispersed in water was measured to be +61 mV. The surface charge of the nanoparticles was reversed by raising the pH of the solution, which was measured to be −43.3 mV at 10.5 pH. In a separate process, the CIGS nanoparticles dispersed in water were coated with PSS. The resulting dispersion was observed to be stable and the surface charge was measured to be −56.9 mV. The LbL deposition process of CIGS nanoparticles was characterized by depositing thin films on quartz crystal microbalance (QCM). LbL depositions was conducted using (i) oppositely charged CIGS nanoparticles, (ii) positively charged CIGS nanoparticles and PSS, and (iii) PSS-coated CIGS (CIGS-PSS) and polyethyleneimine (PEI). The average thickness of each bi-layer of the above mentioned depositions were measured to be 2.2 nm, 1.37 nm, and 10.12 nm, respectively. The results from the QCM have been observed to be consistent with the film thickness results obtained from atomic force microscopy (AFM). Various immersion times versus thickness of the film were also studied. For electrical characterization, the CIGS films were deposited on indium tindioxide (ITO)-coated glass substrates. Current versus voltage (I/V) measurements were carried out for each of the films using the Keithley semiconductor characterization instruments and micromanipulator probing station. It was observed that the conductivity of the films was increased with the deposition of each additional layer. The I/V characteristics were also measured under the light illumination and after annealing to study the photovoltaic and annealing effects. It was observed that under light illumination, the resistivity of a 12-layer CIGS film decreased by 93% to 0.54 MΩ.m, and that of the same number of layers of PSS-coated CIGS and PEI film decreased by 60% to 0.97 MΩ.m under illumination. The resistivity of an 8-layer CIGS and PSS film decreased by 76.4% to 0.1 MΩ.m, and that of the same layers of PSS-coated CIGS and PEI decreased by 87% to 0.07 MΩ.m after annealing. The functionalized nanoparticles and the LbL CIGS films were implemented in the solar cell devices. Several configurations of CIGS films (p-type), and ZnO and CdS films (n-type) were considered. Poly(3,4-ethylenedioxythiophene) (PEDOT), molybdenum (Mo), and ITO were used as back contacts and ITO was used as front contact for all the devices. The devices were characterized the Keithley semiconductor characterization instruments and micromanipulator probing station. For a CIGS and n-ZnO films device with PEDOT as back contact and ITO as front contact, the current density at 0 V and under light illumination was measured to be 60 nA/cm2 and the power density was measured to be 0.018 nW/cm2. For a CIGS and CdS films device with ITO as both back and front contact, the current density at 0 V and under light illumination was measured to be 50 nA/cm2 and the power density was measured to be 0.01 nW/cm2. For a drop-casted CIGS and CdS films device with Mo as back contact and ITO as front contact, the current density of 50 nA/cm2 at 0 V and power density of 0.5 nW/cm2 under light illumination was measured. For the LbL CIGS and chemical bath deposited CdS films device with ITO as both back and front contact, the current density of 0.04 mA/cm2 at 0 V and power density of 1.6 μW/cm2 under light illumination was measured. Comparing to Device-III, an increase by 99% in the power density was observed by using the CIGS LbL film in the device structure. The novel aspects of this research include, (i) functionalization of the CIGS nanoparticles to disperse in water including coating with PSS, (ii) electrostatic LbL deposition of CIGS films using oppositely charged nanoparticles and polymers, and (iii) the utilization of the fabricated LbL CIGS films to develop solar cells. In addition, the n-type cadmium sulfide film (CdS) and zinc oxide (ZnO) buffer layer were also deposited through LbL process after the respective particles were functionalized with PSS coating in separate experiments.Item Performance of Multiple Emission Peak Light Emitting Diode Light Curing Unit: Degree of Conversion and Microhardness of Resin-Based Pit and Fissure Sealant(2022-07) Ba Armah, Ibrahim; Soto, Armando E.; Platt, Jeffrey A.; Al Dehailan, Laila A.Background: The light-cured resin-based pit and fissure sealants success and longevity are enhanced by sufficient curing. Multiple emission peak Light Emitting Diode Light Curing Units offer a wider range of wavelengths and different levels of irradiances to ensure sufficient curing.The irradiance is considered a main curing factor that can affect the material properties. Purpose: The aim of this study was to assess the effect of different settings of a multiwave LED LCU on the degree of conversion and microhardness of a pit and fissure sealant comparing the irradiance of 1000 mW/cm2 to 1400 mW/cm2 and 3200 mW/cm2 irradiances of the LCU using manufacturer’s guidelines for curing times at 2, 4 and, 6 mm distances. Methods: A multiwave LED light curing unit was evaluated on three different irradiance levels 1000 mW/cm2 (S), 1400 mW/cm2 (H), and 3200 mW/cm2 (X). A total of 90 samples made from the fissure sealant were fabricated and divided into eighteen groups (n=5/group). Samples were cured following manufacturer’s guidelines of curing times for each curing mode at 2, 4, or 6 mm distance between the light tip and top of samples. The DC was measured using (ATR-FTIR) spectroscopy. The KHN test was performed on five different locations of each specimen using a hardness tester (Leco LM247AT, MI, USA, software; Confident V 2.5.2). Results: The top DC for H-8 was significantly higher than S-10 at 2 and 4mm, H-20 DC was significantly lower than S-30 at only 2mm. The bottom DC for H-8 was significantly higher than S-10 at 2mm only, H-20 DC was significantly lower than S-30 at 4 and 6mm only. H-8 KHN at top surface was significantly lower than S-10 at 2mm only, H-20 was significantly lower than S-30 at 2 and 6mm only. H-8 KHN at bottom surface was significantly lower than S-10 at 4 and 6mm but significantly higher at 2mm. H-20 was significantly lower than S-30 at 2mm but significantly higher at 4 and 6mm. The top DC for X-3 was significantly lower than S-10 at all curing distances with no significant difference at all curing distances between X-9 and S-30. The bottom DC for X-3 was significantly higher than S-10 at all curing distances with no significant difference between X-9 and S-30. X-3 KHN at top surface was significantly lower than S-10s at all curing distances. X-9 was significantly lower than S-30 at 6mm only. X-3 KHN at bottom surface was significantly lower than S-10 at 2 and 4mm only with no significant difference at all curing distances between X-9 and S-30. Conclusions: Using a multiwave LED LCU to polymerize Delton Opaque resin-based fissure sealants will result in an optimal DC and KHN values for any irradiance level if the curing distance is kept at 4 mm or less and with at least two cycles of the shortest curing time recommended by the manufacturer. Using a multiwave LED LCU with 1000, 1400 or 3200 mW/cm2 irradiance levels with shortest curing times recommended resulted in unsatisfactory DC and KHN levels. LED LCU with high and extra high irradiance levels (1400 and 3200 mW/cm2) can result in high DC and KHN levels when used adequately. Xtra Power mode (3200 mW/cm2) used on shortest curing time (3 seconds) resulted in significantly lower mechanical properties and for that reason it is not recommended to be used.