Ganetespib with Methotrexate Acts Synergistically to Impede NF-κB/p65 Signaling in Human Lung Cancer A549 Cells.

Among the various types of cancer, lung cancer accounts for the highest number of fatalities across the globe. A combination of different cancer chemotherapeutics is regarded as an effective strategy for clinical management of different cancers. Ganetespib (GAN) is a well-established hsp90 inhibitor with enhanced pharmacological properties in comparison with its first-generation counterparts. Previous preclinical studies have shown that GAN exerts significant effects against cancer cells; however, its therapeutic effects against non-small cell lung cancer (NSCLC) A549 cells, achieved by modulating the expression of the NF-κB/p65 signaling pathway, remains unexplored. In this study, the combinatorial effect of GAN and methotrexate (MTX) against lung carcinomas was investigated through both in silico and in vitro studies. A combinatorial treatment regimen of GAN/MTX exerted more significant cytotoxic effects (p < 0.001) against A549 cells than individual treatments. The GAN/MTX combination also instigated nuclear fragmentation followed by augmentation in intracellular ROS levels (p < 0.001). The elevated ROS in A549 cells upon exposure to GAN/MTX combinatorial regimen was concomitantly accompanied with a remarkable reduction in mitochondrial viability. In addition, it was observed that the GAN/MTX combination succeeded in elevating caspase-3 activity and downregulating the expression levels of anti-apoptotic mediators Bcl2 and survivin in NSCLC A549 cells. Most importantly, the GAN/MTX combinatorial regimen impeded the activation of the NF-kB/p65 signaling pathway via repression of the expression of E-cadherin and N-cadherin, which was confirmed by molecular docking studies. Collectively, these findings demonstrated the synergistic effect of the GAN/MTX combinatorial regimen in suppressing the growth of A549 cells by modulating the NF-κB/p65 signaling pathway.

Integrated breeding approaches to enhance the nutritional quality of food legumes.

Global food security, both in terms of quantity and quality remains as a challenge with the increasing population. In parallel, micronutrient deficiency in the human diet leads to malnutrition and several health-related problems collectively known as "hidden hunger" more prominent in developing countries around the globe. Biofortification is a potential tool to fortify grain legumes with micronutrients to mitigate the food and nutritional security of the ever-increasing population. Anti-nutritional factors like phytates, raffinose (RFO's), oxalates, tannin, etc. have adverse effects on human health upon consumption. Reduction of the anti-nutritional factors or preventing their accumulation offers opportunity for enhancing the intake of legumes in diet besides increasing the bioavailability of micronutrients. Integrated breeding methods are routinely being used to exploit the available genetic variability for micronutrients through modern "omic" technologies such as genomics, transcriptomics, ionomics, and metabolomics for developing biofortified grain legumes. Molecular mechanism of Fe/Zn uptake, phytate, and raffinose family oligosaccharides (RFOs) biosynthesis pathways have been elucidated. Transgenic, microRNAs and genome editing tools hold great promise for designing nutrient-dense and anti-nutrient-free grain legumes. In this review, we present the recent efforts toward manipulation of genes/QTLs regulating biofortification and Anti-nutrient accumulation in legumes using genetics-, genomics-, microRNA-, and genome editing-based approaches. We also discuss the success stories in legumes enrichment and recent advances in development of low Anti-nutrient lines. We hope that these emerging tools and techniques will expedite the efforts to develop micronutrient dense legume crop varieties devoid of Anti-nutritional factors that will serve to address the challenges like malnutrition and hidden hunger.

Epigenomics as Potential Tools for Enhancing Magnitude of Breeding Approaches for Developing Climate Resilient Chickpea.

Epigenomics has become a significant research interest at a time when rapid environmental changes are occurring. Epigenetic mechanisms mainly result from systems like DNA methylation, histone modification, and RNA interference. Epigenetic mechanisms are gaining importance in classical genetics, developmental biology, molecular biology, cancer biology, epidemiology, and evolution. Epigenetic mechanisms play important role in the action and interaction of plant genes during development, and also have an impact on classical plant breeding programs, inclusive of novel variation, single plant heritability, hybrid vigor, plant-environment interactions, stress tolerance, and performance stability. The epigenetics and epigenomics may be significant for crop adaptability and pliability to ambient alterations, directing to the creation of stout climate-resilient elegant crop cultivars. In this review, we have summarized recent progress made in understanding the epigenetic mechanisms in plant responses to biotic and abiotic stresses and have also tried to provide the ways for the efficient utilization of epigenomic mechanisms in developing climate-resilient crop cultivars, especially in chickpea, and other legume crops.

Mapping QTLs for Alternaria blight in Linseed (Linum usitatissimum L.).

A SSR-based linkage map of linseed constructed based on 154 individual lines of F 2 mapping population derived from JRF-4 (disease-resistant) and Chambal (disease susceptible) genotypes. QTLs for Alternaria blight and other yield related traits identified. Out of 1720 SSRs, 216 SSRs were found polymorphic among the parents but due to segregation distortion 18 SSRs could not be used for linkage map construction. Total 191 SSRs were used to construct the linkage map and distributed in 15 linkage groups covering genome length of 1802.4 cM. A total of 10 QTLs were identified for 4 phenotypic traits including 4 QTLs for capsules/plant, 2 for capsule weight/plant, 2 for seed weight/plant and 2 for Alternaria blight resistance. This study laid a foundation for further validation and fine mapping with more advance and large set of marker for different QTL identification and marker-assisted selection in linseed.

Estimation of genetic diversity and population structure in Tinospora cordifolia using SSR markers.

Thirty polymorphic SSRs, derived from RNA sequencing of Tinospora cordifolia (willd.), were utilized for genetic diversity and population structure evaluation among 96 accessions collected from ten different geographical regions of India. A total of 7611 SSRs were identified from 268149 transcripts. Of all SSR loci, 69.07% of them were tri-nucleotide repeat motifs, followed by di-nucleotide repeat motifs (12.82%). A total of 230 alleles were generated by 30 SSRs with an average of 7.67 alleles per locus with comparatively higher polymorphic information content (average 0.68). The expected (He) and observed (Ho) heterozygosity means were 0.71 and 0.12, respectively. All the loci showed significant deviation from Hardy-Weinberg Equilibrium (HWE). The neighbor joining clustering based on jaccard's coefficient grouped all the 96 accessions into three major cluster which was also in congruence with model-based structure plot. The result of molecular variance (AMOVA) revealed higher genetic variance within populations than among populations. The result reflects an existence of high level of genetic diversity in the collected accessions of T. cordifolia. The accessions Tc131, Tc31, Tc129, Tc38, Tc16, Tc59, Tc60, Tc17, Tc106 and Tc130 was found to be potential and diverse in nature and the SSRs TCSSR-18, TCSSR-37, TCTSSR-59, TCTSSR-92, TCTSSR-123 and TCTSSR-126 as potential markers. These accessions and newly developed SSR markers provide valuable resource and could be strategically utilized for further genetic improvement of T. cordifolia.

Mapping QTLs for 15 morpho-metric traits in Arabidopsis thaliana using Col-0 × Don-0 population.

Genome wide quantitative trait loci (QTL) mapping was conducted in Arabidopsis thaliana using F2 mapping population (Col-0 × Don-0) and SNPs markers. A total of five linkage groups were obtained with number of SNPs varying from 45 to 59 per linkage group. The composite interval mapping detected a total of 36 QTLs for 15 traits and the number of QTLs ranged from one (root length, root dry biomass, cauline leaf width, number of internodes and internode distance) to seven (for bolting days). The range of phenotypic variance explained (PVE) and logarithm of the odds ratio of these 36 QTLs was found be 0.19-38.17% and 3.0-6.26 respectively. Further, the epistatic interaction detected one main effect QTL and four epistatic QTLs. Five major QTLs viz. Qbd.nbri.4.3, Qfd.nbri.4.2, Qrdm.nbri.5.1, Qncl.nbri.2.2, Qtd.nbri.4.1 with PVE > 15.0% might be useful for fine mapping to identify genes associated with respective traits, and also for development of specialized population through marker assisted selection. The identification of additive and dominant effect QTLs and desirable alleles of each of above mentioned traits would also be important for future research.

Genetic diversity and population structure analysis of Indian garlic (Allium sativum L.) collection using SSR markers.

Genetic diversity was assessed among 53 Indian garlic accessions using SSR markers. Initially, 24 SSR primer pairs were used for screening three selected garlic accessions. Out of 24 SSR primer pairs, 10 primer pairs which consistently showed good amplification and polymorphism were selected for DNA profiling. SSR primer pairs showed PIC values ranging from 0.30 to 0.99. Based on AMOVA we found that the greater part of the genetic diversity was expected due to intra population with 84% variation and only 16% of variation was due to among populations suggesting presence of genetic structure. The results of cluster analysis and principal component analysis largely correspond to each other. Population structure analysis revealed genetic differentiation of accessions. The results of present study revealed existence of significant variability in Indian garlic germplasm.

Assessment of genetic diversity in Lepidium sativum L. using inter simple sequence repeat (ISSR) marker.

Lepidium sativum L. is a fast-growing, edible and medicinal plant that belongs to the family Brassicaceae. Indian system of medicinal and health (ISHM) recognizes this plant as a source of several medicinal and nutraceutical factors. Ninety-four accessions collected from 19 states of India were assessed for genetic diversity using inter-simple sequence repeat (ISSR) marker. Ten ISSR primers amplified a total of 172 bands across the 94 accessions and out of these, 139 bands were found to be polymorphic and 33 as monomorphic. The percentage polymorphism varied from 60.00 to 91.30% with an average of 80.10%. The polymorphism information content (PIC) varied from 0.14 to 0.39 with an average of 0.27. The Jaccard similarity coefficient ranged from 0.11 to 0.89 with minimum between accession LS61 and LS60 and maximum between accession LS95 and LS81. Cluster analysis based on UPGMA grouped all the 94 accessions into three major clusters with accessions per cluster ranging from 12 to 45. Similar to UPGMA clustering, PCA also differentiated all the accessions into three major groups. Model-based clustering determined three sub-populations (K = 3). Further, analysis of molecular variance showed that 67% of allelic diversity was attributed to individual accessions within populations while 33% was distributed among populations. This preliminary study shows that significant variability exists in the collected accessions.

Assessment of genetic diversity and population structure in gladiolus (Gladiolus hybridus Hort.) by ISSR markers.

ISSR (Inter simple sequence repeat) markers were used to assess the genetic diversity and population structure in 53 indigenous and exotic genotypes of gladiolus (Gladiolus hybridus Hort.). Molecular markers analysis showed PIC ranges from 0.42 (ISSR 861) to 0.99 (ISSR 855, ISSR 856 and ISSR 889) with an average 0.812, marker index ranged from 0.99 (ISSR 889) to 9.26 (ISSR 851) with an average 4.66 and resolving power of the primers ranged from 0.03 (ISSR 889) to 11.58 (ISSR 861) with an average value 3.80. The dendrogram based UPGMA clustering showed that all the 53 genotypes grouped into three main clusters. Nei's gene diversity (Na) varied from 0.929 to 1.717, effective number of alleles (Ne) varied from 1.262 to 1.369, Shannon's information index (I) ranged from 0.251 to 0.359 and gene diversity (He) was in the range from 0.167 to 0.229. Population structure analysis revealed three groups in which 32 genotypes were admixture types.

A Scanning Electron Microscopic Evaluation of the Effectiveness of Etidronic Acid, SmearClear and MTAD in Removing the Intracanal Smear Layer.

STATEMENT OF THE PROBLEM: Root canal therapy should not simply be the extirpation of the pulp and widening of the canal. But one should also focus on how to completely remove the loosely-attached smear layer because it has adverse effects on the final outcome of the treatment. PURPOSE: This study compared the efficacy of Etidronic acid, SmearClear and MTAD to remove the smear layer created during instrumentation in different regions of the root canal. MATERIALS AND METHOD: Fifty single-rooted mandibular premolars were decoronated from the cementoenamel junction and instrumented using the ProTaper universal rotary file system along with copious irrigation by 1.0% sodium hypochlorite and distilled water. On the basis of the type of chelating agent used for irrigation, samples (n=10) were then randomized into five groups as: Group I- 9% etidronic acid, Group II- 18% etidronic acid, Group III- SmearClear, Group IV- MTAD and Group V- normal saline. Subsequent to irrigation, all samples were rinsed, dried and sectioned longitudinally for evaluation of the smear layer removal under scanning electron microscope (2000X). Data were statistically analyzed by two-way analysis of variance and Tukey's post hoc test with statistical significance set at p< 0.5. RESULTS: The result showed that SmearClear was the most efficient in removing the smear layer. However, etidronic acid was found inferior than both SmearClear and MTAD. CONCLUSION: Chelators are essential for complete smear layer removal in association with organic solvent.

Genetic diversity, population structure and association analysis in linseed (Linum usitatissimum L.).

The present investigation aimed to explore the level of genetic diversity, determine the population structure in a larger set of germplasm of linseed using microsatellite marker and identify linked markers through association mapping. A total of 168 accessions of linseed were evaluated for major agro-economic traits and SSRs markers deployed for diversity assessment. A total of 337 alleles were amplified by 50 SSRs ranging from 2 to 13 with an average of 6.74 ± 2.8 alleles per loci. The neighbor joining based clustering grouped all the accessions into three major clusters that were also confirmed by scatter plot of PCoA. While model based clustering determined four sub-populations (K = 4). Further, analysis of molecular variance analysis considering three population showed that maximum variation (79%) was within the population. We identified one putative SSR marker (Lu_3043) linked with days to 50% flowering through both GLM and MLM analysis of association mapping. The results of this preliminary study revealed genetic diversity, population structure in linseed and linked marker which could be utilized in future breeding program.

Endodontic Management of a Maxillary First Molar with Seven Root Canals Using Spiral Computed Tomography.

The main objective of this case report is to present a rare root canal configuration of maxillary molar with seven root canals; three mesiobuccal, two palatal and two distobuccal canals diagnosed during treatment procedure confirmed by spiral computed tomography. A thorough knowledge of root canal morphology, proper clinical and radiographic examination, and use of dental operating microscopes are necessary for successful clinical outcomes. This article highlights the variations in the morphology of maxillary first molar and use of the latest techniques in successful diagnosis and negotiation of the additional canals.

The effectiveness of eucalyptus oil, orange oil, and xylene in dissolving different endodontic sealers.

OBJECTIVE: The objective of this study was to evaluate the dissolution effectiveness of eucalyptus oil, orange oil, xylene, and distilled water on three different endodontic sealers. MATERIALS AND METHODS: About 240 samples of root canal sealers (eighty for each sealer) were prepared and divided into four groups of 20 each for immersion in different organic solvents. Each group was further subdivided into two subgroups (n = 10) for 2 and 10 min of immersion time. The mean percentage of weight loss was determined for each sealer in each solvent at both time periods. Data were statistically analyzed by two factor analysis of variance and significance of mean difference was obtained by Tukey's post hoc test (P < 0.05). RESULTS: The lowest level of solubility was observed for Adseal followed by Apexit Plus and Endomethasone N at both time periods in all solvents. Apexit Plus showed no significant (P > 0.05) difference in its dissolution in all the organic solvents except distilled water at both the time periods. The solubility profile of Endomethasone N and Adseal did not differ significantly among eucalyptus oil, orange oil, and xylene at 2 min and between eucalyptus oil and orange oil at 10 min. However, at 10 min, Endomethasone N and Adseal showed a more pronounced solubility in xylene as compared to both eucalyptus oil and orange oil. CONCLUSIONS: In general, xylene was the most effective in dissolving root canal sealers than other organic solvents. Essential oils (eucalyptus oil and orange oil) were found similar in their ability to dissolve Apexit Plus and Endomethasone N.

Accidental aspiration/ingestion of foreign bodies in dentistry: A clinical and legal perspective.

The potential of foreign body aspiration or ingestion is a worldwide health problem in dentistry. The general dental practitioners should be extremely attentive in handling of minor instruments during any intervention related to the oral cavity, especially in the supine or semi-recumbent position of the patient. Aspiration cases are usually more critical and less common than ingestion. We report a case of iatrogenic aspiration of an endodontic broach, which gets disclosed during the recording of past dental history of the patient. The patient was asymptomatic during that time. A quick posterior-anterior chest radiograph was taken which revealed the presence of broach in the lower lobe of the left lung. The patient was immediately referred to the pulmonary medicine department where the fiberoptic bronchoscope retrieval was planned, and the same was carried out successfully under local anesthesia. Although such accidents have rare occurrence, the associated risks and morbidity are too high to be overlooked, especially from the viewpoint of special care, resources, and the associated financial cost required for their management. Moreover, practitioners are also liable for malpractice litigation given the fact that such cases are avoidable. This article also discusses relevant review literature, risk factors, symptoms, and management of such iatrogenic accidents along with drawing attention to the significance of preventive measures and their role in avoiding meritorious legal and ethical issues.

Efficacy of etidronic acid, BioPure MTAD and SmearClear in removing calcium ions from the root canal: An in vitro study.

OBJECTIVE: The purpose of this study was to quantify the amount of calcium ions removed from the root canal by etidronic acid (HEBP), BioPure MTAD, and SmearClear using atomic absorption spectrophotometer. MATERIALS AND METHODS: Fifty (n = 50) freshly extracted human mandibular premolar teeth were collected and decoronated at the cementoenamel junction. The canals were prepared in a crown down fashion using the rotary system and copiously irrigated with 1.0% sodium hypochlorite. All specimens were rinsed with the deionized water. Based on the type of chelating agent used, the samples (n = 10) were randomly divided into five (four test and one negative control) groups. Accordingly, Group I - 9% HEBP, Group II - 18% HEBP, Group III - SmearClear, Group IV - BioPure MTAD, and Group V - normal Saline. Subsequent to irrigation, the solution was collected in a test tube and subjected to atomic absorption spectrophotometer for the quantification of calcium ions removed from the root canal. RESULTS: The mean concentration of calcium ions removed from the root canal (mean ± standard deviation) in all groups (I-V) were 13.32 ± 0.54 µg/ml, 16.36 ± 0.27 µg/ml, 20.04 ± 0.24 µg/ml, 18.15 ± 0.39 µg/ml, and 8.74 ± 0.49 µg/ml, respectively. CONCLUSIONS: SmearClear was the most effective agent for the removal of calcium ions from the root canal. Hence, its combined use with an organic solvent can be recommended for efficient smear layer removal.

Diversity analysis in Indian genotypes of linseed (Linum usitatissimum L.) using AFLP markers.

AFLP fingerprinting of 45 Indian genotypes of linseed was carried out to determine the genetic relationship among them. Sixteen primer combinations produced 1142 fragments with 1129 as polymorphic and 13 as monomorphic fragments. Polymorphic fragments varied from 44 (E-ACA/M-CTA) to 94 (E-AGC/M-CAC) with an average of 70.6 fragments per primer combination. The frequency of polymorphism varied from 93.7% to 100% with an average of 98.8% across all the genotypes. The PIC value ranged from 0.19 to 0.31 with an average of 0.23 per primer combination. The primer pair E-AGC/M-CAC showed the maximum PIC value (0.31) followed by E-AGC/M-CAG (0.29), E-AAC/M-CAG (0.26) and E-AGC/M-CTA (0.25). Resolving power (RP) and marker index (MI) varied from 13.73 to 43.50 and 8.81 to 28.91 respectively. The Jaccard's similarity coefficient varied from 0.16 to 0.57 with an average of 0.26 ± 0.05. The maximum genetic similarities (57%) were detected between genotypes Him Alsi-1 and Him Alsi-2, followed by Him Alsi-1 and GS41 and GS41 and LC-54. The genotypes R-552, Himani, RKY-14, Meera, Indira Alsi-32 and Suyog were found to be more divergent genotypes. The NJ clustering grouped all the 45 genotypes into three major clusters. In general the genotypes of cluster III had high oil content and those of cluster I had low oil content. At the population level, within population variance was much higher than between populations variance.

Trends in management of myofacial pain.

We systematically reviewed the myofascial pain publications in the literature. The aim of this article is to review the methods of management and their outcome and factors associated with prognosis. The topics of interest in the diagnostic process are myofascial trigger points electromyography, jaw tracking, joint sound recorder, sonography, and vibratography, exclusion of other orofacial pain and temporomandibular disorders. Management modalities are occlusal therapy, physiotherapy, multidimensional rehabilitation antinociceptive therapy, anti-inflammatory and analgesics, muscle relaxants, stretch, and spray technique, transcutaneous electric nerve stimulation, and in severe cases botulinum toxin may be tried. The disease required interdisciplinary interaction in terms of occlusal therapy, antinociceptive therapy and physiotherapy because management of the disease may be influenced by the specialist primarily treating the patients.

Large-scale resource development in Gossypium hirsutum L. by 454 sequencing of genic-enriched libraries from six diverse genotypes.

The sequence information has been proved to be an essential genomic resource in case of crop plants for their genetic improvement and better utilization by humans. To dissect the Gossypium hirsutum genome for large-scale development of genomic resources, we adopted hypomethylated restriction-based genomic enrichment strategy to sequence six diverse genotypes. Approximately 5.2-Gb data (more than 18.36 million reads) was generated which, after assembly, represents nearly 1.27-Gb genomic sequences. We predicted a total of 93,363 gene models (21,399 full length) and identified 35,923 gene models which were validated against already sequenced plant genomes. A total of 1,093 transcription factor-encoding genes, 3,135 promoter sequences and 78 miRNA (including 17 newly identified in Gossypium) were predicted. We identified significant no. of molecular markers including 47,093 novel simple sequence repeats and 66,364 novel single nucleotide polymorphisms. In addition, we developed NBRI-Comprehensive Cotton Genomics database, a web resource to provide access of cotton-related genomic resources developed at NBRI. This study contributes considerable amount of genomic resources and suggests a potential role of genic-enriched sequencing in genomic resource development for orphan crop plants.

Development and characterization of genomic and expressed SSRs for levant cotton (Gossypium herbaceum L.).

Four microsatellite-enriched genomic libraries for CA(15), GA(15), AAG(8) and ATG(8) repeats and transcriptome sequences of five cDNA libraries of Gossypium herbaceum were explored to develop simple sequence repeat (SSR) markers. A total of 428 unique clones from repeat enriched genomic libraries were mined for 584 genomic SSRs (gSSRs). In addition, 99,780 unigenes from transcriptome sequencing were explored for 8,900 SSR containing sequences with 12,471 expressed SSRs. The present study adds 1,970 expressed SSRs and 263 gSSRs to the public domain for the use of genetic studies of cotton. When 150 gSSRs and 50 expressed SSRs were tested on a panel of four species of cotton, 68 gSSRs and 12 expressed SSRs revealed polymorphism. These 200 SSRs were further deployed on 15 genotypes of levant cotton for the genetic diversity assessment. This is the first report on the successful use of repeat enriched genomic library and expressed sequence database for microsatellite markers development in G. herbaceum.