Comparison of Root End Sealing Ability of Three Retrograde Filling Materials in Teeth with Root Apices Resected at 90° using Dye Penetration Method under Fluorescent Microscope: An In-vitro Study

Background: Inadequate seal at the apex is the substantial cause for surgical endodontic dissatisfaction. The retrograde filling material which is used should prevent the egress of potential contaminants into periradicular tissue. Objectives: To evaluate the ability of MTA Angelus, Zirconomer and Bioactive bone cement to seal the root end as retrograde filling material; and to compare root end sealing ability of these three different retrograde filling materials with apices resected at 900 angles using dye penetration method under fluorescent microscope. Methodology: Thirty six extracted upper anterior teeth are to be cut horizontally at the CEJ. After following the standard protocols of, “cleaning, shaping and obturation” with gutta percha and “AH Plus sealer”, the samples will be resected 3mm at the apical end at 90 degree angle along the long axis of the tooth with the help of diamond disc. A root end cavity of depth 3mm will be made with Diamond coated ultrasonic surgical tip S12 90 ND. The teeth will be randomly categorized in 3 groups: Group 1: MTA angelus; Group 2: Zirconomer; Group 3: Bioactive Bone Cement. Following which the roots to be coated with nail varnish except the tip. Each material will be compressed in the root end cavity with the help of small pluggers. All the samples which are retrofilled will be kept in acrydine orange for a duration of 24 hours, following which cleaning & bucco-lingual sectioning(vertical) will be done. Fluorescent microscope will be used for observation of sectioned root samples. Expected Results: Bioactive bone cement is expected to have better sealing ability of the retrograde cavity preparation with minimal or no microleakage followed by MTA Angelus and then Zirconomer. Conclusion: If this study proves correct, this would be helpful for the clinicians to choose better and the most efficient retrograde filling material with best sealing ability and minimal microleakage in a retrograde preparation which will aid in success of the root canal treatment further resolving the infection.

Application of Transposon Systems in The Transgenesis of Bovine Somatic And Germ Cells

Background: Several DNA transposons, PiggyBac (PB), Sleeping beauty (SB) and Tol2 have been applied as effective means for transgenesis in many species. Cattle are not typical experimental animals, and relatively little verification has been studied in this species. Thus, the goal of this study was the applicability of three transposon systems in somatic and embryo cells in cattle, while also determining which of the three systems is appropriate for each type of cell. To conduct the experiment, green fluorescent protein (GFP)-expressing transposon systems were used for electroporation and microinjection in the somatic cells and embryo stage, respectively. After transfection, GFP-positive cells or blastocysts were observed through a fluorescent microscope and transfection efficiency was calculated by FACS.Results: In the bovine somatic cells experiment, the PB (63.97 ± 11.56) showed higher efficiency as compared to the other two systems (SB: 50.74 ± 13.02 and Tol2: 16.55 ± 5.96). Unlike the results of the somatic cells, Tol2 (75.00%) and SB (70.00%) in the embryo were more efficient as compared to PB (42.86 %).Conclusions: These results demonstrate that all three transposon systems can be used in bovine somatic cells and embryos as a gene engineering experimental method and which type of transposon system is appropriate to apply depending on the cell type.

Attenuation of Quorum Sensing Controlled Virulence Factors and Biofilm Formation by Edible Fruit Extract of Coccinia indica against Pseudomonas aeruginosa

Background: Pseudomonas aeruginosa is a Gram-negative opportunistic human pathogen that mainly infects immunocompromised individuals and patients with urinary tract infection and chronic infections of the respiratory pathways, including cystic fibrosis. Many quorum sensing (QS) controlled components such as bio surfactants and swarming motilities play an important role in the establishment of biofilms. Targeting these factors through anti-QS strategies prevent biofilm formation and treating infections. Coccinia indica commonly called little gourd is used to treat diabetes, wound, burn infections and has antioxidant, antibacterial and antitussive properties. Methods: The methanolic fruit extract of C. indica was prepared and screened for anti-QS and anti-biofilm formation activity. Pyocyanin inhibition, rhamnolipid, crystal violet staining assay tests was performed and the extract was observed under fluorescent microscope. Results: The results obtained are as follows - the fruit extract inhibits the pyocyanin at 58.13% and 42.27% at 0.5 mg/ml and 1.0 mg/ml, biofilm at 69.86% and 49.06% at 0.5 mg/ml and 1.0 mg/ml, inhibits rhamnolipid assay and under fluorescent microscope it is seen scattered whereas control produce biofilm matrix like appearance. Conclusion: Since less study has been made on the quorum sensing and biofilm activity of C.indica our study aimed to fulfil it and it was found that it exhibits good biofilm formation and thus can be used for treating infections.

Suppression of Ripe Rot on ‘Zesy002’ Kiwifruit with Commercial Agrochemicals

Ripe rot caused by Botryosphaeria dothidea is one of the serious diseases of postharvest kiwifruit. In order to control ripe rot on Actinidia chinensis cultivar ‘Zesy002’, several commercial agrofungicides were selected by an antifungal test on an artificial medium. Furthermore, disease suppression by the selected fungicides was evaluated on the kiwifruit by inoculation with a conidial suspension of B. dothidea. On the artificial media containing boscalid + fludioxonil was shown to be the most effective antifungal activity. However, in the bio-test pyraclostrobin + boscalid and iminoctadine-tris were the most effective agrochemicals on the fruit. On the other hand, the infection structures of B. dothidea on kiwifruit treated with pyraclostrobin + boscalid were observed with a fluorescent microscope. Most of the fungal conidia had not germinated on the kiwifruit treated with the agrochemicals whereas on the untreated fruit the fungal conidia had mostly germinated. Electron microscopy of the fine structures showed morphological changes to the conidia and branch of hyphae on the kiwifruit pre-treated with pyraclostrobin + boscalid, indicating its suppression effect on fungal growth. Based on this observation, it is suggested that ripe rot by B. dothidea may be suppressed through the inhibition of conidial germination on the kiwifruit treated with the agrochemicals.

Phototoxicities Caused by Continuous Light Exposure Were Not Induced in Retinal Ganglion Cells Transduced by an Optogenetic Gene

The death of photoreceptor cells is induced by continuous light exposure. However, it is unclear whether light damage was induced in retinal ganglion cells with photosensitivity by transduction of optogenetic genes. In this study, we evaluated the phototoxicities of continuous light exposure on retinal ganglion cells after transduction of the optogenetic gene mVChR1 using an adeno-associated virus vector. Rats were exposed to continuous light for a week, and visually evoked potentials (VEPs) were recorded. The intensities of continuous light (500, 1000, 3000, and 5000 lx) increased substantially after VEP recordings. After the final recording of VEPs, retinal ganglion cells (RGCs) were retrogradely labeled with a fluorescein tracer, FluoroGold, and the number of retinal ganglion cells was counted under a fluorescent microscope. There was no significant reduction in the amplitudes of VEPs and the number of RGCs after exposure to any light intensity. These results indicated that RGCs were photosensitive after the transduction of optogenetic genes and did not induce any phototoxicity by continuous light exposure.

Eye Protection in ENT Practice During the COVID-19 Pandemic

Objectives There is a lack of evidence-based guidelines with regard to eye protection for aerosol-generating procedures in otolaryngology practice. In addition, some recommended personal protective equipment (PPE) is not compatible with commonly used ENT equipment. This study aims to investigate the degree of eye protection that commonly used PPE gives. Study Design Simulation model. Setting Simulation laboratory. Methods A custom-built setup was utilized to simulate the clinical scenario of a patient cough in proximity of a health care worker. A system that sprays a xanthan-fluorescein mixture was set up and calibrated to simulate a human cough. A mannequin with cellulose paper placed on its forehead, eyes, and mouth was fitted with various PPE combinations and exposed to the simulated cough. The degree of contamination on the cellulose papers was quantified with a fluorescent microscope able to detect aerosols ≥10 µm. Results When no eye protection was worn, 278 droplets/aerosols reached the eye area. The use of the surgical mask with an attached upward-facing shield alone resulted in only 2 droplets/aerosols reaching the eye area. In this experiment, safety glasses and goggles performed equally, as the addition of either brought the number of droplets/aerosols reaching the eye down to 0. Conclusion When used with an upward-facing face shield, there was no difference in the eye protection rendered by safety goggles or glasses in this study. Safety glasses may be considered a viable alternative to safety goggles in aerosol-generating procedures.

The Surface Antigen 1(SAG1) of Toxoplasma Gondii Interacts With RACK1 Inducing Autophagy and Maintaining The Viability of Host Cells

BackgroundIt has been well known that the surface antigen 1(SAG 1) of T.gondii plays an important role in the invasion of Tachyzoite into host cells. However whether it also play a role in the intracellular parasitism of T.gondii remains unclear. The main purpose of this study was to determine the effect of SAG1 on host cells and investigate the underlying mechanism. MethodsSAG1 was overexpressed in human embryonic kidney cell 293(HEK) by transfection. Autophagy was determined by fluorescent microscope and flow cytometry (FCM) in HEK293 cells co-transfected with Flag-SAG1 and EGFP-LC3. The interaction of SAG1 and RACK1 was measured by co-immunoprecipitation(Co-IP), GST pulldown and fluorescent microscope. The expression of cytokines including IL-1β, IL-6, and IL-12 was determined by qRT-PCR. The expression of LC3, Ki67 and RACK1 was detected by Western blot. Cellular senescence was measured by β-galactosidase staining.ResultsWe found that overexpression of SAG1 in human embryonic kidney cells (HEK293) induces non-canonical autophagy and inhibition of autophagy using hydroxychloroquine (HCQ) significantly decreases the cell viability of HEK293 cells. Mechanically, we identified RACK1, an intracellular multifunctional protein as a binding partner of SAG1. Depletion of RACK1 inhibited SAG1 induces non-autophagy and decreases the enhanced expression of cytokines including IL-1β, IL-6, IL-12 and TNF-α in SAG1 overexpressing cells. ConclusionThese data showed that SAG1 could induce non-canonical autophagy and facilitate the expression of IL-1β, IL-6, IL-12 and TNF-α by interacting with RACK1, maintaining the viability of host cells. Our results suggests a new contribution of SAG1 in the intracellular parasitism.

A modular microscopic smartphone attachment for imaging and quantification of multiple fluorescent probes using machine learning

A smartphone-based fluorescent microscope capable of imaging multiple fluorescent probes at different magnification levels.