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.

Studies on Almond Gum and Gelucire-Based Pellets Prepared by Extrusion and Spheronization for Sustained Release.

Objectives: The aim of the investigation was to prepare sustained release (SR) pellets of diltiazem hydrochloride employing almond gum and gelucire. The study was performed to explore the suitability of almond gum in the preparation of pellets of diltiazem hydrochloride without the use of microcrystalline cellulose and role and effectiveness of hydrophobic gelucire (43/01) in controlling the drug release. Materials and Methods: Pellets were prepared by extrusion-spheronization of the blend previously obtained by incorporation of the drug in a mixture of melted gelucire 43/01 and almond gum. A 32 factorial design was employed to study the effect of two independent variables, almond gum and gelucire, on the size, friability and drug release from pellets. Scanning electron microscopy, differential scanning calorimetry and infrared spectroscopy were performed to characterize pellets. Results: Free flowing spherical pellets could be prepared. The 32 factorial study revealed that as the proportion of almond gum increased, the size of pellets increased, while increasing gelucire had opposite effect. The yield of pellets prepared in different formulations is in the range of 86 to 92%. The size of the pellets varied from 1128 to 1458 µ. Higher amounts of gelucire resulted in pellets with greater friability, whereas increasing the amount of almond gum yielded pellets with low friability. The pellets exhibited SR of diltiazem and the presence of gelucire in the matrix of the pellets had an enhanced sustaining effect on release. Conclusion: Dispersion of the drug in gelucire before it was converted to pellets resulted in extended release of drug. The drug release rate changed with changes in the proportion of pellet composition. The results of the study suggest that employing gelucire (43/01) in the preparation of pellets is a useful approach in the design of SR products of highly water-soluble drug such as diltiazem hydrochloride.

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.

Enhancement of Vancomycin Potential against Pathogenic Bacterial Strains via Gold Nano-Formulations: A Nano-Antibiotic Approach.

The remarkable rise of antibiotic resistance among pathogenic bacteria poses a significant threat to human health. Nanoparticles (NPs) have recently emerged as novel strategies for conquering fatal bacterial diseases. Furthermore, antibiotic-functionalized metallic NPs represent a viable nano-platform for combating bacterial resistance. In this study, we present the use of vancomycin-functionalized gold nanoparticles (V-GNPs) to battle pathogenic bacterial strains. A facile one-pot method was adopted to synthesize vancomycin-loaded GNPs in which the reducing properties of vancomycin were exploited to produce V-GNPs from gold ions. UV-Visible spectroscopy verified the production of V-GNPs via the existence of a surface plasmon resonance peak at 524 nm, whereas transmission electron microscopy depicted a size of ~24 nm. Further, dynamic light scattering (DLS) estimated the hydrodynamic diameter as 77 nm. The stability of V-GNPs was investigated using zeta-potential measurements, and the zeta potential of V-GNPs was found to be -18 mV. Fourier transform infrared spectroscopy confirmed the efficient loading of vancomycin onto GNP surfaces; however, the loading efficiency of vancomycin onto V-GNPs was 86.2%. Finally, in vitro antibacterial studies revealed that V-GNPs were much more effective, even at lower concentrations, than pure vancomycin. The observed antibacterial activities of V-GNPs were 1.4-, 1.6-, 1.8-, and 1.6-fold higher against Gram-negative Escherichia coli, Klebsiella oxytoca, and Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus, respectively, compared to pure vancomycin. Collectively, V-GNPs represented a more viable alternative to pure vancomycin, even at a lower antibiotic dose, in conquering pathogenic bacteria.

Assigning secondary structure in proteins using AI.

Knowledge about protein structure assignment enriches the structural and functional understanding of proteins. Accurate and reliable structure assignment data is crucial for secondary structure prediction systems. Since the 1980s, various methods based on hydrogen bond analysis and atomic coordinate geometry, followed by machine learning, have been employed in protein structure assignment. However, the assignment process becomes challenging when missing atoms are present in the protein files. Our method proposed a multi-class classifier program named DLFSA for assigning protein secondary structure elements (SSE) using convolutional neural networks (CNNs). A fast and efficient GPU-based parallel procedure extracts fragments from protein files. The model implemented in this work is trained with a subset of the protein fragments and achieves 88.1% and 82.5% train and test accuracy, respectively. The model uses only Cα coordinates for secondary structure assignments. The model has been successfully tested on a few full-length proteins also. Results from the fragment-based studies demonstrate the feasibility of applying deep learning solutions for structure assignment problems.

Current Status of Brain Tumor in the Kingdom of Saudi Arabia and Application of Nanobiotechnology for Its Treatment: A Comprehensive Review.

OBJECTIVE: Brain tumors are the most challenging of all tumors and accounts for about 3% of all cancer allied deaths. The aim of the present review is to examine the brain tumor prevalence and treatment modalities available in the Kingdom of Saudi Arabia. It also provides a comprehensive analysis of the application of various nanotechnology-based products for brain cancer treatments along with their prospective future advancements. METHODS: A literature review was performed to identify and summarize the current status of brain cancer in Saudi Arabia and the scope of nanobiotechnology in its treatment. RESULTS: Depending upon the study population data analysis, gliomas, astrocytoma, meningioma, and metastatic cancer have a higher incidence rate in Saudi Arabia than in other countries, and are mostly treated in accordance with conventional treatment modalities for brain cancer. Due to the poor prognosis of cancer, it has an average survival rate of 2 years. Conventional therapy includes surgery, radiotherapy, chemotherapy, and a combination thereof, but these do not control the disease's recurrence. Among the various nanomaterials discussed, liposomes and polymeric nanoformulations have demonstrated encouraging outcomes for facilitated brain cancer treatment. CONCLUSIONS: Nanomaterials possess the capacity to overcome the shortcomings of conventional therapies. Polymer-based nanomaterials have shown encouraging outcomes against brain cancer when amalgamated with other nano-based therapies. Nonetheless, nanomaterials could be devised that possess minimal toxicity towards normal cells or that specifically target tumor cells. In addition, rigorous clinical investigations are warranted to prepare them as an efficient and safe modality for brain cancer therapy.

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.

Corpus callosum dermoid cyst: A rare entity.

Dermoid cyst is a congenital lesion that arises due to embryogenic impairment. It accounts for less than 0.3% of all intracranial masses. Herein we report a 30-year-old male who presented with a short history of headache and behavioral disorder. Physical and radiological examination diagnosed it to be a lipoma/epidermoid arising from corpus callosum. The lesion was removed endoscopically and histopathologic examination was performed which confirmed it to be a dermoid cyst. Rarity of the lesion and difficulty in pre-operative diagnosis prompted us to bring forward this case report.

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.