Effect of Irrigation of Sodium Hypochlorite at Different Temperatures and Concentration on the Elimination of Enterococcus Faecalis from Rootcanals: An In Vitro Study

Sixty single rooted anterior teeth were collected, cleaned and stored in normal saline. They were standardized to a length of 15mm and the root canals preparation was done using Protaper instruments as per the manufacturer’s instructions. The teeth were then divided into groups and inoculated with Enterococcus faecalis. Followed by irrigation with varying concentrations of Sodium hypochlorite (1%, 3% and 5%) at temperatures of (240Celsius, 370Celsius,450Celsius). Sampling was done by inserting paper points followed by culturing. After 48hrs of incubation typical Enterococcus faecalis colonies were counted in all plates, results tabulated and statistically analyzed. CONCLUSION: On completion of the study on the effect of varying concentration of sodium hypochlorite at three different temperatures on Enterococcus faecalis within the canals. The following conclusions were made: 1. 5% sodium hypochlorite at 450Celsius was found to be the most effective irrigant used. 2. Statistical analysis with ANOVA at 5% level of significance showed there is a significant difference between 5% sodium hypochlorite at 450Celsius and other two groups of 5% sodium hypochlorite at 370Celsius and 240Celsius which means that 5%sodium hypochlorite at 450 is better. 3. Based on the results of the study chair side pre-warming sodium hypochlorite before use with a temperature controlled device is recommended. 4. 5% Sodium hypochlorite at 240Celsius and 370Celsius were also able to inhibit the growth of Enterococcus faecalis efficiently in comparison to the control group. Effectiveness of sodium hypochlorite as an irrigant at lower concentration could be enhanced by increasing the temperature. 5. The effect of varying concentrations of sodium hypochlorite as an irrigant at varying temperature in a clinical setting should be evaluated

PREDICTION OF ANTI-PARKINSON POTENTIAL OF PHYTOCONSTITUENTS USING PREDICTION OF ACTIVITY SPECTRA OF SUBSTANCES SOFTWARE

Objective: Neurodegenerative disorders are group of diseased conditions in which there is loss of neuron cells occur. The main objective of this study to find/search out the phytochemical with the help of prediction of activity spectra of substances (PASSs), those show maximum activity over the selected targets of the Parkinson's disease (PD).Methods: PASSs is a valuable software which is used in this study, to predict the anti-Parkinson activity of different compounds. Canonical simplified molecular-input line-entry system is used for the prediction of anti-Parkinson activity which is obtained from PubChem website. The predicted activity also compared with marketed compound like levodopa.Results: From the study, it was found that resveratrol was the only compound which has the activity on all the selected targets. On the other hand, stemazole and celastrol were found to have the least active compounds as both have the activity only on a single target.Conclusion: In this research work, we tried to compile the information regarding the PASS predicted anti-Parkinson activity of some important phytoconstituents. We found that resveratrol can be a target for further investigation in the development of drug therapy for PD

Preparation of highly and generally enriched mammalian tissues for solid state NMR.

An appreciable level of isotope labelling is essential for future NMR structure elucidation of mammalian biomaterials, which are either poorly expressed, or unexpressable, using micro-organisms. We present a detailed protocol for high level (13)C enrichment even in slow turnover murine biomaterials (fur keratin), using a customized diet supplemented with commercial labelled algal hydrolysate and formulated as a gel to minimize wastage, which female mice consumed during pregnancy and lactation. This procedure produced approximately eightfold higher fur keratin labelling in pups, exposed in utero and throughout life to label, than in adults exposed for the same period, showing both the effectiveness, and necessity, of this approach.The authors would like to acknowledge funding from the Biotechnology and Biological Sciences Research Council for DGR and RR; Engineering and Physical Sciences Research Council for WYC and VWCW; National Institute of Health Research for RAB.This is the final version of the article. It first appeared from Springer via http://dx.doi.org/10.1007/s10858-015-9977-

Collagen labelling with an azide-proline chemical reporter in live cells.

We have developed a strategy for selective imaging of collagen in live foetal ovine osteoblasts. Our approach involves the incorporation of an azide-tagged proline in the biosynthesis of collagen followed by labelling using a strain-promoted [3+2] azide-alkyne cycloaddition reaction

Time to Turn on the Blue Lights: A Systematic Review and Meta-analysis of Photodynamic Diagnosis for Bladder Cancer.

CONTEXT: White light (WL) cystoscopy and transurethral resection of bladder tumour (TURBT) comprise the current gold standard technique for detecting and grading bladder cancer. However, with WL cystoscopy, recurrence following initial TURBT is high, and identification of smaller tumours and carcinoma in situ is poor. Photodynamic diagnosis (PDD) has been developed to improve the detection of bladder. OBJECTIVE: To assess the effect of PDD-guided TURBT compared with WL on recurrence rates (RRs) in non-muscle-invasive bladder cancer (NMIBC). EVIDENCE ACQUISITION: A systematic review of the literature from inception to April 2020 using Medline, EMBASE, and CENTRAL was undertaken. Randomised control trials comparing TURBT undertaken with PDD to WL that reported RRs of at least 12 mo were included in the analysis. The primary outcomes were RRs at 12 and 24 mo. The secondary outcomes were reported adverse effects. The Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology was used to assess the certainty of the evidence. EVIDENCE SYNTHESIS: Twelve randomised controlled trials (2288 patients) were included for the meta-analysis. PDD was found to reduce RRs at 12 mo (RR 0.73, confidence interval [CI] 0.60-0.88) and 24 mo (RR 0.75, CI 0.62-0.91). There was an increased risk of recurrence for patients undergoing WL at 12 mo (hazard ratio [HR] 1.14, CI 1.05-1.23) and 24 mo (HR 1.25, CI 1.15-1.35). Two studies reported recurrence data at 60 mo showing statistically significant outcomes in favour of PDD: one showed lower RRs for PDD (49% PDD vs 68% WL), whilst the other showed increased recurrence-free survival (68.2% PDD vs 57.3% WL). Adverse effects appeared to be minimal, though poorly reported. A GRADE analysis showed the evidence to be of moderate certainty overall. CONCLUSIONS: This systematic review found that PDD reduced RRs and improved recurrence-free survival compared with WL in NMIBC over at least 2-yr follow-up. These effects may persist up to 5 yr. Further research in a pragmatic study looking at longer-term outcomes beyond 24 mo will help guide recommendations on clinical adoption. PATIENT SUMMARY: This review suggests that photodynamic diagnosis, compared with white light cystoscopy, improves recurrence-free survival in non-muscle-invasive bladder cancer over at least 2 yr of follow-up. However, confirmatory pragmatic studies with longer-term outcomes are required for its clinical adoption

Proline provides site-specific flexibility for in vivo collagen.

Fibrillar collagens have mechanical and biological roles, providing tissues with both tensile strength and cell binding sites which allow molecular interactions with cell-surface receptors such as integrins. A key question is: how do collagens allow tissue flexibility whilst maintaining well-defined ligand binding sites? Here we show that proline residues in collagen glycine-proline-hydroxyproline (Gly-Pro-Hyp) triplets provide local conformational flexibility, which in turn confers well-defined, low energy molecular compression-extension and bending, by employing two-dimensional 13C-13C correlation NMR spectroscopy on 13C-labelled intact ex vivo bone and in vitro osteoblast extracellular matrix. We also find that the positions of Gly-Pro-Hyp triplets are highly conserved between animal species, and are spatially clustered in the currently-accepted model of molecular ordering in collagen type I fibrils. We propose that the Gly-Pro-Hyp triplets in fibrillar collagens provide fibril "expansion joints" to maintain molecular ordering within the fibril, thereby preserving the structural integrity of ligand binding sites.BBSRC, EPSRC, Raymond and Beverly Sackler Fund for Physics of Medicine, Wellcome Trust, ER

NMR spectroscopy of native and in vitro tissues implicates polyADP ribose in biomineralization.

Nuclear magnetic resonance (NMR) spectroscopy is useful to determine molecular structure in tissues grown in vitro only if their fidelity, relative to native tissue, can be established. Here, we use multidimensional NMR spectra of animal and in vitro model tissues as fingerprints of their respective molecular structures, allowing us to compare the intact tissues at atomic length scales. To obtain spectra from animal tissues, we developed a heavy mouse enriched by about 20% in the NMR-active isotopes carbon-13 and nitrogen-15. The resulting spectra allowed us to refine an in vitro model of developing bone and to probe its detailed structure. The identification of an unexpected molecule, poly(adenosine diphosphate ribose), that may be implicated in calcification of the bone matrix, illustrates the analytical power of this approach

Mapping quantitative trait loci (QTLs) associated with resistance to major pathotype-isolates of pearl millet downy mildew pathogen

Downy mildew (DM) caused by Sclerospora graminicola is the most devastating disease of pearl millet. It may lead to annual grain yield losses of up to ~80% and substantial deterioration of forage quality and production. The present study reports construction of the linkage map integrating simple sequence repeat (SSR) markers, for detection of quantitative trait loci (QTLs) associated withDMresistance in pearl millet. Amapping population comprising of 187 F8 recombinant inbred lines (RILs) was developed from the cross (ICMB 89111-P6 × ICMB 90111-P6). The RILs were evaluated for disease reaction at a juvenile stage in the greenhouse trials. Genotyping data was generated from 88 SSR markers on RILs and used to construct genetic linkage map comprising of 53 loci on seven linkage groups (LGs) spanning a total length of 903.8 cM with an average adjacent marker distance of 18.1 cM. Linkage group 1 (LG1; 241.1 cM) was found to be longest and LG3 the shortest (23.0 cM) in length. The constructed linkage map was used to detect five large effect QTLs for resistance to three different pathotype-isolates of S. graminicola from Gujarat (Sg445), Haryana (Sg519) and Rajasthan (Sg526) states of India. One QTL was detected for isolate Sg445 resistance, and two each for Sg519 and Sg526 resistance on LG4 with LOD scores ranging from 5.1 to 16.0, explaining a wide range (16.7% to 78.0%) of the phenotypic variation (R2). All the five co-localized QTLs on LG4 associated with the DM resistance to the three pathotype-isolates were contributed by the resistant parent ICMB 90111-P6. The QTLs reported here may be useful for the breeding programs aiming to develop DM resistant pearl millet cultivars with other desirable traits using genomic selection (GS) approaches

Classification of Clinical Isolates of Klebsiella pneumoniae Based on Their in vitro Biofilm Forming Capabilities and Elucidation of the Biofilm Matrix Chemistry With Special Reference to the Protein Content

Klebsiella pneumoniae is a human pathogen, capable of forming biofilms on abiotic and biotic surfaces. The limitations of the therapeutic options against Klebsiella pneumoniae is actually due to its innate capabilities to form biofilm and harboring determinants of multidrug resistance. We utilized a newer approach for classification of biofilm producing Klebsiella pneumoniae isolates and subsequently we evaluated the chemistry of its slime, more accurately its biofilm. We extracted and determined the amount of polysaccharides and proteins from representative bacterial biofilms. The spatial distribution of sugars and proteins were then investigated in the biofilm matrix using confocal laser scanning microscopy (CLSM). Thereafter, the extracted matrix components were subjected to sophisticated analysis incorporating Fourier transform infrared (FTIR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, one-dimensional gel-based electrophoresis (SDS-PAGE), high performance liquid chromatography (HPLC), and MALDI MS/MS analysis. Besides, the quantification of its total proteins, total sugars, uronates, total acetyl content was also done. Results suggest sugars are not the only/major constituent of its biofilms. The proteins were harvested and subjected to SDS-PAGE which revealed various common and unique protein bands. The common band was excised and analyzed by HPLC. MALDI MS/MS results of this common protein band indicated the presence of different proteins within the biofilm. The 55 different proteins were identified including both cytosolic and membrane proteins. About 22 proteins were related to protein synthesis and processing while 15 proteins were identified related to virulence. Similarly, proteins related to energy and metabolism were 8 and those related to capsule and cell wall synthesis were 4. These results will improve our understanding of Klebsiella biofilm composition and will further help us design better strategies for controlling its biofilm such as techniques focused on weakening/targeting certain portions of the slime which is the most common building block of the biofilm matrix