NanoLuc Bioluminescence-Driven Photodynamic Activation of Cholecystokinin 1 Receptor with Genetically-Encoded Protein Photosensitizer MiniSOG

In contrast to reversible activation by agonist, cholecystokinin 1 receptor (CCK1R) is permanently activated by singlet oxygen generated in photodynamic action, with sulphonated aluminium phthalocyanine or genetically encoded mini singlet oxygen generator (miniSOG) as photosensitizer. In these works, a halogen light source was used to power photodynamic action. For possible in vivo application of photodynamic CCK1R physiology, bearing a cumbersome light-delivery device connected to an external light source by experimental animals might interfere with their behavior. Therefore, in the present work, the possibility of bioluminescence-driven miniSOG photodynamic CCK1R activation was examined, as monitored by Fura-2 calcium imaging. In parallel experiments, it was found that, after plasma membrane (PM)-localized expression of miniSOGPM in AR4-2J cells, light irradiation with blue light-emitting diode (LED) (450 nm, 85 mW·cm−2, 1.5 min) induced persistent calcium oscillations that were blocked by CCK1R antagonist devazepide 2 nM. NanoLuc was expressed bicistronically with miniSOGPM via an internal ribosome entry site (IRES) sequence (pminiSOGPM-IRES-NanoLuc). The resultant miniSOGPM-IRES-NanoLuc-AR4-2J cells were found to generate strong bioluminescence upon addition of NanoLuc substrate coelenterazine. Strikingly, coelenterazine 5 microM was found to trigger long-lasting calcium oscillations (a hallmark for permanent CCK1R activation) in perifused miniSOGPM-IRES-NanoLuc-AR4-2J cells. These data indicate that NanoLuc bioluminescence can drive miniSOGPM photodynamic CCK1R activation, laying the foundation for its future in vivo applications.

Evaluation of biochemical changes in dental tissues after different office bleaching methods

The color of the teeth is an important topic for many people and can be influenced by intrinsic and extrinsic stains. There is an increasing demand for whitening of the teeth year by year. The most popular way of whitening is “bleaching,” which is the result of the breakdown of pigments located in the enamel and/or the dentin, caused by reactive oxygen species (ROS) released from bleaching agents. These bleaching agents could increase matrix metalloproteinase (MMP)-mediated collagen degradation in dentin. The aim of this study was to compare biochemical changes and oxidative stress levels of the human premolar dentin–pulp complex after three different bleaching methods containing hydrogen peroxide (H2O2) bleaching agents. Individuals, whose first four premolars were extracted for orthodontic purposes, included into the study. Group 1—Laser: bleaching gel containing 46% H2O2 (LaserWhite20 whitening gel, Biolase Technology Inc., San Clemente, CA, USA) and a diode laser activation (Ezlase 940 nm system). Group 2: 35% H2O2 containing whitening gel (Whiteness HP Maxx, FGM) and halogen light source activation (Optilux 501, Kerr, Orange, CA, USA). Group 3: 35% H2O2 containing whitening gel (Whiteness HP Maxx, FGM). Group 4—Control: No whitening treatment. According to the test results, there were no significant differences among groups in the values of cathepsin B and MMP proteolytic activities ( p > 0.05). The total ROS values released from the dentin tissue were higher than those obtained from the pulp tissue ( p < 0.05). There were significant differences among the bleaching groups in the ROS values released from the dentin tissue.

PENGARUH WAKTU PERENDAMAN TiO2 DALAM LARUTAN EKSTRAK ANTOSIANIN BUNGA ROSELLA PADA KINERJA DYE SENSITIZED SOLAR CELL (DSSC)

The use of solar panels is still quite expensive in the making, for it needs to be developed solar panels are cheap based DSSC (Dye Sensitized Solar Cell). The use of DSSC is very good developed in Indonesia which is famous for its biological richness. This study used rosella flowers as photosensitizer obtained from the extraction process in the form of maceration for 24 hours. The components used to construct a DSSC outline can be divided into four parts, namely substrate, dye, semiconductor titanium dioxide (TiO2), and electrolyte. In this research the Ti O2 layer was immersed in a solution of rosella flower extract with different time variations, ie 1 hour, 2 hours, and 3 hours. Light sources used are sunlight and 150 watt halogen lamps. It was found that at 3 hours of immersion using the sunlight source was the best result of maximally current (Imax) 0.3 mA, maximum voltage (V) 582,4 mV and its efficiency 0,258%. The results obtained were lower when the DSSC was given a halogen light source, i.e. Maximum current (Imax) 0.08 mA, maximal voltage (Vmax) 125,6 mV and its efficiency 0,167%. The greater the intensity of the light illumination the greater the output power of the DSSC.The use of solar panels is still quite expensive in the making, for it needs to be developed solar panels are cheap based DSSC (Dye Sensitized Solar Cell). The use of DSSC is very good developed in Indonesia which is famous for its biological richness. This study used rosella flowers as photosensitizer obtained from the extraction process in the form of maceration for 24 hours. The components used to construct a DSSC outline can be divided into four parts, namely substrate, dye, semiconductor titanium dioxide (TiO2), and electrolyte. In this research the TiO2 layer was immersed in a solution of rosella flower extract with different time variations, ie 1 hour, 2 hours, and 3 hours. Light sources used are sunlight and 150 watt halogen lamps. It was found that at 3 hours of immersion using the sunlight source was the best result of maximally current (Imax) 0.3 mA, maximum voltage (Vmax) 582,4 mV and its efficiency 0,258%. The results obtained were lower when the DSSC was given a halogen light source, i.e. Maximum current (Imax) 0.08 mA, maximal voltage (Vmax) 125,6 mV and its efficiency 0,167%. The greater the intensity of the light illumination the greater the output power of the DSSC.

NPK DETECTION SPECTROSCOPY ON NON-AGRICULTURE SOIL

Soil is a medium for plant roots to grow, absorb water and necessary solutes for growth. Soil macronutrient testing is helpful for determining the nutrients content in soil before applying fertilizer for quality and process controls of agricultural produce and soil fertility. Spectroscopy is an emerging technology which is rapid and simple has been widely used in agricultural and food analysis processes. The capability of spectroscopy to characterize material from the transmission or absorbance has been used in this paper to measure nitrogen (N), phosphorus (P) and potassium (K) content in non-agriculture soil. The paper details preliminary characterization of soil spectroscopy with a Deuterium-Halogen light source and Ocean Optic spectrometer to measure the absorbance level of the macronutrients. The extracted nutrients were mixed with the colour reagent and specific colored solution was developed. Two soil samples have been employed for the experimental characterization, which are mud flood and kaolin. The result shows that high absorbance level of N at 450 nm in wavelength, P at 750 nm for both samples. The absorbance level of K was measured high at 500nm for mud flood and 450nm for kaolin. In addition, the tested macronutrients give similar wavelength of peak absorbance level at 970 nm for both samples. For future works, the optical measurements will be implemented using visible and near infrared LED and the photodetector in order to replace the spectrometer usage for soil spectroscopy. This would lead to achieve the primary objective of this research in developing a simple and low cost spectroscopy uses light-emitting diode (LED).

Effect of halogene light source on marginal microleakage of resin composite restorations

Introduction. Although the technology of production resin composite materials has been improved in the past years, polymerization shrinkage and microleakage still remain the main problem of the resin composite restorations. The most important issues are polymerization control, light source choice, quality and polymerization technique. The aim of this study was to evaluate the effect of the halogen light source on the marginal microleakage of the resin composite restorations class V. Material and Methods. The study included 40 extracted human teeth (20 intact and 20 with caries lesion). Class V cavity was prepared in each tooth. The light curing unit used in this study had irradiance 540 mW/cm2. Cavities were restored with the two resin composite materials, Filtek Supreme (3M ESPE) and Luksogal - Galenika. Microleakage was determined using colored solution of 50% silver-nitrate. Results were obtained using stereomicroscope with 6 times magnification. Color penetration was evaluated on the occlusal and gingival wall. Results. Linear color penetration was observed in all cavities restored using Luksogal. There was significant difference (p<0.01) in color penetration between the occlusal (20.30 ?m) and gingival wall (34.00 ?m). Color penetration was shown in all cavities prepared on intact and caries teeth and restored using Filtek Supreme. Also, color penetration on the occlusal wall (13.80 ?m) was significantly lower (p<0.01) than the penetration on the gingival wall (33.00 ?m). Conclusion. All the tested cavities showed marginal leakage, regardless the tooth or material used. Greater microleakage was noticed when Luksogal was used.

Microleakage under Orthodontic Brackets Using High-Intensity Curing Lights

Objective: To compare the microleakage of the enamel-adhesive-bracket complex at the occlusal and gingival margins of brackets bonded with high-intensity light curing lights and conventional halogen lights. Materials and Methods: Forty-five freshly extracted human maxillary premolar teeth were randomly separated into three groups of 15 teeth each. Stainless steel brackets were bonded in all groups according to the manufacturer's recommendations. Specimens (15 per group) were cured for 40 seconds with a conventional halogen light, 20 seconds with light-emitting diode (LED), and 6 seconds with plasma arc curing light (PAC). After curing, the specimens were further sealed with nail varnish, stained with 0.5% basic-fuchsine for 24 hours, sectioned and examined under a stereomicroscope, and scored for microleakage for the enamel-adhesive and bracket-adhesive interfaces from both the occlusal and gingival margins. Statistical analyses were performed using Kruskal-Wallis and Mann-Whitney U-tests with a Bonferroni correction. Results: The type of light curing unit did not significantly affect the amount of microleakage at the gingival or occlusal margins of investigated interfaces (P &gt;.05). The gingival sides in the LED and PAC groups exhibited higher microleakage scores compared with those observed on occlusal sides for the enamel-adhesive and adhesive-bracket interfaces. The halogen light source showed similar microleakage at the gingival and occlusal sides between both adhesive interfaces. Conclusions: High-intensity curing units did not cause more microleakage than conventional halogen lights. This supports the use of all these curing units in routine orthodontic practice.

Optical mapping of Langendorff-perfused human hearts: establishing a model for the study of ventricular fibrillation in humans

Our objective was to establish a novel model for the study of ventricular fibrillation (VF) in humans. We adopted the established techniques of optical mapping to human ventricles for the first time to determine whether human VF is the result of wave breaks and singularity point formation and is maintained by high-frequency rotors and fibrillatory conduction. We describe the technique of acquiring optical signals in human hearts during VF, their characteristics, and the feasibility of possible analyses that could be performed to elucidate mechanisms of human VF. We used explanted hearts from five cardiomyopathic patients who underwent transplantation. The hearts were Langendorff perfused with Tyrode solution (95% O2-5% CO2), and the potentiometric dye di-4-ANEPPS was injected as a bolus into the coronary circulation. Fluorescence was excited at 531 ± 20 nm with a 150-W halogen light source; the emission signal was long-pass filtered at 610 nm and recorded with a mapping camera. Fractional change of fluorescence varied between 2% and 12%. Average signal-to-noise ratio was 40 dB. The mean velocity of VF wave fronts was 0.25 ± 0.04 m/s. Submillimetric spatial resolution (0.65–0.85 mm), activation mapping, and transformation of the data to phase-based analysis revealed reentrant, colliding, and fractionating wave fronts in human VF. On many occasions the VF wave fronts were as large as the entire vertical length (8 cm) of the mapping field, suggesting that there are a limited number of wave fronts on the human heart during VF. Phase transformation of the optical signals allowed the first demonstration ever of phase singularity point, wave breaks, and rotor formation in human VF. This method provides opportunities for potential analyses toward elucidation of the mechanisms of VF and defibrillation in humans.