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Introduction: Treatment of mechanical exposure of the pulp during caries excavation presents a clinical challenge. In this case series of 15 patients, with a follow-up period of over a year, the outcome of direct pulp capping with Biodentine (septodont) after mechanical pulp exposure was assessed. Aim of Study: The aim of this study is to evaluate the outcome of direct pulp capping with Biodentine in deeply carious teeth when pulp was mechanically exposed during caries excavation and cavity preparation. Vital pulps exposed during caries excavation in molar teeth were treated with 3% sodium hypochlorite for 2 min. If adequate hemostasis was achieved, the pulp tissue was capped with Biodentine, which covered the entire pulpal floor. This was followed by the placement of a layer of resin-modified glass ionomer cement and a final layer of composite resin (Filtek Z350-3M) to complete the restoration. The patients were recalled periodically and evaluated for any evidence of pulpal/periapical pathology. Results: In the follow-up period that ranged from 12 to 24 months, all teeth were asymptomatic. Conclusion: Biodentine appears to be a suitable material for direct pulp capping under clinical conditions. However, long-term follow-up studies and controlled trials involving a large sample size are warranted.

Endodontic treatments are routinely done with the help of radiographs. However, radiographs represent only a two-dimensional image of an object. Failure to identify aberrant anatomy can lead to endodontic failure. This case report presents the use of three-dimensional imaging with cone beam computed tomography (CBCT) as an adjunct to digital radiography in identification and management of mandibular second molar with three mesial canals.

Complex root canal system with atypical variations is a common finding among mandibular premolars. Endodontic treatment in these teeth may not be successful due to the failure to recognise and treat multiple canals. This paper presents endodontic treatment of a mandibular second premolar with three roots and three canals.

Heat shock transcription factors (HSFs) contribute significantly to thermotolerance acclimation. Here, we identified and cloned a putative HSF gene (HSFA2h) of 1218 nucleotide (acc. no. KP257297.1) from wheat cv. HD2985 using a de novo transcriptomic approach and predicted sHSP as its potential target. The expression of HSFA2h and its target gene (HSP17) was observed at the maximum level in leaf tissue under heat stress (HS), as compared to the control. The HSFA2h-pRI101 binary construct was mobilized in Arabidopsis, and further screening of T3 transgenic lines showed improved tolerance at an HS of 38 °C compared with wild type (WT). The expression of HSFA2h was observed to be 2.9- to 3.7-fold higher in different Arabidopsis transgenic lines under HS. HSFA2h and its target gene transcripts (HSP18.2 in the case of Arabidopsis) were observed to be abundant in transgenic Arabidopsis plants under HS. We observed a positive correlation between the expression of HSFA2h and HSP18.2 under HS. Evaluation of transgenic lines using different physio-biochemical traits linked with thermotolerance showed better performance of HS-treated transgenic Arabidopsis plants compared with WT. There is a need to further characterize the gene regulatory network (GRN) of HSFA2h and sHSP in order to modulate the HS tolerance of wheat and other agriculturally important crops.

For many years, the impact of Particulate Matter (PM) in the ambient air has been one of the major concerns for the environment and human health. The consideration of the heterogeneity and complexity of different size fractions is notably important for the assessment of PM toxicological effects. The aim of the study was to present a comprehensive size-composition-morphology characterization and to assess the oxidative potential, genotoxicity, and mutagenicity of the atmospheric PM fractions, collected by using MOUDI near a busy roadside in Lucknow, India. Physicochemical characterization of ambient coarse particles (1.8–10 µm), fine particles (0.32–1.8 µm), quasi-ultrafine (0.1–0.32 µm) and ultrafine particles (≤0.1 µm) along with SRM 1649b was done using TEM, SEM, DLS, NTA, ICP-MS, and IC in parallel with the estimation of exogenous Reactive Oxygen Species (ROS) by acellular assays. In this study, two different acellular assays, dithiothreitol (DTT) and the CM-H2DCFDA assay, indicated stronger mass-normalized bioactivity for different size ranges. Enrichment factor analysis indicated that the different size fractions were highly enriched with elements of anthropogenic origin as compared to elements of crustal origin. The endotoxin concentration in different size fractions was also estimated. Cellular studies demonstrated significant uptake, cytotoxicity, ultrastructural changes, cellular ROS generation, and changes in the different phases of the cell cycle (Sub G1, G1, S, G2/M) exposed to different size fractions. The Comet assay and the Micronucleus assay were used to estimate genotoxicity. Mutagenic potential was revealed by the HGPRT gene forward mutation assay in V-97 cells. Conclusively, our results clearly indicate that the genotoxic and mutagenic potential of the coarse PM was greater than the other fractions, and interestingly, the ultrafine PM has higher bioactivity as compared to quasi-ultrafine PM.

Wheat, being sensitive to terminal heat, causes drastic reduction in grain quality and yield. MAPK cascade regulates the network of defense mechanism operated inside plant system. Here, we have identified 21 novel MAPKs through gel-based proteomics and RNA-seq data analysis. Based on digital gene expression, two transcripts (transcript_2834 and transcript_8242) showing homology with MAPK were cloned and characterized from wheat (acc. nos. MK854806 and KT835664). Transcript_2834 was cloned in pET28a vector and recombinant MAPK protein of ∼40.3 kDa was isolated and characterized to have very high in-vitro kinase activity under HS. Native MAPK showed positive correlation with the expression of TFs, HSPs, genes linked with antioxidant enzyme (SOD, CAT, GPX), photosynthesis and starch biosynthesis pathways in wheat under HS. Wheat cv. HD3086 (thermotolerant) having higher expression and activity of MAPK under HS showed significant increase in accumulation of proline, H2O2, starch, and granule integrity, compared with BT-Schomburgk (thermosusceptible).

Wheat is highly prone to terminal heat stress (HS) under late-sown conditions. Delayed- sowing is one of the preferred methods to screen the genotypes for thermotolerance under open field conditions. We investigated the effect of terminal HS on the thermotolerance of four popular genotypes of wheat i.e. WR544, HD2967, HD2932, and HD2285 under field condition. We observed significant variations in the biochemical parameters like protein content, antioxidant activity, proline and total reducing sugar content in leaf, stem, and spike under normal (26 ± 2°C) and terminal HS (36 ± 2°C) conditions. Maximum protein, sugars and proline was observed in HD2967, as compared to other cultivars under terminal HS. Wheat cv. HD2967 showed more adaptability to the terminal HS. Differential protein-profiling in leaves, stem and spike of HD2967 under normal (26 ± 2°C) and terminal HS (36 ± 2°C) showed expression of some unique protein spots. MALDI-TOF/MS analysis showed the DEPs as RuBisCO (Rub), RuBisCO activase (Rca), oxygen evolving enhancer protein (OEEP), hypothetical proteins, etc. Expression analysis of genes associated with photosynthesis (Rub and Rca) and starch biosynthesis pathway (AGPase, SSS and SBE) showed significant variations in the expression under terminal HS. HD2967 showed better performance, as compared to other cultivars under terminal HS. SSS activity observed in HD2967 showed more stability under terminal HS, as compared with other cultivars. Triggering of different biochemical parameters in response to terminal HS was observed to modulate the plasticity of carbon assimilatory pathway. The identified DEPs will enrich the proteomic resources of wheat and will provide a potential biochemical marker for screening wheat germplasm for thermotolerance. The model hypothesized will help the researchers to work in a more focused way to develop terminal heat tolerant wheat without compromising with the quality and quantity of grains.

Heat stress is one of the major problems in agriculturally important cereal crops, especially wheat. Here, we have constructed a subtracted cDNA library from the endosperm of HS-treated (42°C for 2 h) wheat cv. HD2985 by suppression subtractive hybridization (SSH). We identified ~550 recombinant clones ranging from 200 to 500 bp with an average size of 300 bp. Sanger’s sequencing was performed with 205 positive clones to generate the differentially expressed sequence tags (ESTs). Most of the ESTs were observed to be localized on the long arm of chromosome 2A and associated with heat stress tolerance and metabolic pathways. Identified ESTs were BLAST search using Ensemble, TriFLD and TIGR databases and the predicted CDS were translated and aligned with the protein sequences available in pfam and InterProScan 5 databases to predict the differentially expressed proteins (DEPs). We observed eight different types of post-translational modifications (PTMs) in the DEPs corresponds to the cloned ESTs—147 sites with phosphorylation, 21 sites with sumoylation, 237 with palmitoylation, 96 sites with S-nitrosylation, 3066 calpain cleavage sites, and 103 tyrosine nitration sites, predicted to sense the heat stress and regulate the expression of stress genes. Twelve DEPs were observed to have transmembrane helixes (TMH) in their structure, predicted to play the role of sensors of HS. Quantitative Real-Time PCR of randomly selected ESTs showed very high relative expression of HSP17 under HS; up-regulation was observed more in wheat cv. HD2985 (thermotolerant), as compared to HD2329 (thermosusceptible) during grain-filling. The abundance of transcripts was further validated through northern blot analysis. The ESTs and their corresponding DEPs can be used as molecular marker for screening or targeted precision breeding program. PTMs identified in the DEPs can be used to elucidate the thermotolerance mechanism of wheat – a novel step towards the development of ‘climate-smart’ wheat.

RuBisCo activase (Rca) is a catalytic chaperone involved in modulating the activity of RuBisCo (key enzyme of photosynthetic pathway). Here, we identified eight novel transcripts from wheat through data mining predicted to be Rca and cloned a transcript of 1.4 kb from cv. HD2985, named as TaRca1 (GenBank acc. no. KC776912). Single copy number of TaRca1 was observed in wheat genome. Expression analysis in diverse wheat genotypes (HD2985, Halna, PBW621 and HD2329) showed very high relative expression of TaRca1 in Halna under control and HS-treated, as compared to other cultivars at different stages of growth. TaRca1 protein was predicted to be chloroplast-localized with numerous potential phosphorylation sites. Nothern blot analysis showed maximum accumulation of TaRca1 transcript in thermotolerant cv. during mealy-ripe stage, as compared to thermosusceptible. Decrease in the photosynthetic parameters was observed in all the cultivars, except PBW621 in response to HS. We observed significant increase in the Rca activity in all the cultivars under HS at different stages of growth. HS causes decrease in the RuBisCo activity; maximum reduction was observed during pollination stage in thermosusceptible cvs. as validated through immunoblotting. We observed uniform carbon distribution in different tissues of thermotolerant cvs., as compared to thermosusceptible. Similarly, tolerance level of leaf was observed maximum in Halna having high Rca activity under HS. A positive correlation was observed between the transcript and activity of TaRca1 in HS-treated Halna. Similarly, TaRca1 enzyme showed positive correlation with the activity of RuBisCo. There is, however, need to manipulate the thermal stability of TaRca1 enzyme through protein engineering for sustaining the photosynthetic rate under HS – a novel approach towards development of ‘climate-smart’ crop.