NOTCH1 mutations have been reported to occur in 10% to 15% of head and neck squamous cell carcinomas (HNSCC). To determine the significance of these mutations, we embarked upon a comprehensive study ...of NOTCH signaling in a cohort of 44 HNSCC tumors and 25 normal mucosal samples through a set of expression, copy number, methylation, and mutation analyses. Copy number increases were identified in NOTCH pathway genes, including the NOTCH ligand JAG1. Gene set analysis defined a differential expression of the NOTCH signaling pathway in HNSCC relative to normal tissues. Analysis of individual pathway-related genes revealed overexpression of ligands JAG1 and JAG2 and receptor NOTCH3. In 32% of the HNSCC examined, activation of the downstream NOTCH effectors HES1/HEY1 was documented. Notably, exomic sequencing identified 5 novel inactivating NOTCH1 mutations in 4 of the 37 tumors analyzed, with none of these tumors exhibiting HES1/HEY1 overexpression. Our results revealed a bimodal pattern of NOTCH pathway alterations in HNSCC, with a smaller subset exhibiting inactivating NOTCH1 receptor mutations but a larger subset exhibiting other NOTCH1 pathway alterations, including increases in expression or gene copy number of the receptor or ligands as well as downstream pathway activation. Our results imply that therapies that target the NOTCH pathway may be more widely suitable for HNSCC treatment than appreciated currently.
is a neurological disorder that affects millions of people worldwide. Anti-epileptic drugs (AEDs) are critical for their management. However, the therapeutic window is narrow, and traditional ...laboratory-based therapeutic drug monitoring (TDM) methods can be time consuming and unsuitable for point-of-care testing. To address this issue, we developed a disposable sensor chip based on molecularly imprinted polymer-modified carbon paste electrodes (MIP-CPs) for the TDM of AEDs such as phenobarbital (PB), carbamazepine (CBZ), and levetiracetam (LEV). In this work, functional monomers (methacrylic acid) and crosslinking monomers (methylene bisacrylamide and ethylene glycol dimethacrylate) were copolymerized in the presence of the AED template and grafted on the graphite particles by simple radical photopolymerization. The grafted particles were mixed with silicon oil, dissolving ferrocene as a redox marker to make the MIP-carbon paste (CP). Disposable sensor chips were fabricated by packing the MIP-CP into the base made of poly (ethylene glycol terephthalate) (PET) film. The sensor's sensitivity was determined using differential pulse voltammetry (DPV), carried out on a single sensor chip for each operation. Linearity was obtained from 0-60 μg/mL in PB and LEV and 0-12 μg/mL in CBZ, covering their respective therapeutic range. The time taken for each measurement was around 2 min. The experiment using whole bovine blood and bovine plasma indicated that the existence of species that interfered had a negligible effect on the test's sensitivity. This disposable MIP sensor provides a promising approach for point-of-care testing and facilitating the management of epilepsy. Compared with existing tests, this sensor offers a faster and more accurate way to monitor AEDs, which is crucial for optimizing therapy and improving patient outcomes. Overall, the proposed disposable sensor chip based on MIP-CPs represents a significant advancement in AED monitoring, with the potential for rapid, accurate, and convenient point-of-care testing.
Non small cell lung cancer (NSCLC) is the leading cause of cancer-related mortality world-wide and the majority of cases are diagnosed at late stages of disease. There is currently no cost-effective ...screening test for NSCLC, and the development of such a test is a public health imperative. Recent studies have suggested that chest computed tomography screening of patients at high risk of lung cancer can increase survival from disease, however, the cost effectiveness of such screening has not been established. In this Phase I/II biomarker study we examined the feasibility of using serum miRNA as biomarkers of NSCLC using RT-qPCR to examine the expression of 180 miRNAs in sera from 30 treatment naive NSCLC patients and 20 healthy controls. Receiver operating characteristic curves (ROC) and area under the curve were used to identify differentially expressed miRNA pairs that could distinguish NSCLC from healthy controls. Selected miRNA candidates were further validated in sera from an additional 55 NSCLC patients and 75 healthy controls. Examination of miRNA expression levels in serum from a multi-institutional cohort of 50 subjects (30 NSCLC patients and 20 healthy controls) identified differentially expressed miRNAs. A combination of two differentially expressed miRNAs miR-15b and miR-27b, was able to discriminate NSCLC from healthy controls with sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of 100% in the training set. Upon further testing on additional 130 subjects (55 NSCLC and 75 healthy controls), this miRNA pair predicted NSCLC with a specificity of 84% (95% CI 0.73-0.91), sensitivity of 100% (95% CI; 0.93-1.0), NPV of 100%, and PPV of 82%. These data provide evidence that serum miRNAs have the potential to be sensitive, cost-effective biomarkers for the early detection of NSCLC. Further testing in a Phase III biomarker study in is necessary for validation of these results.
Implementation of highly sophisticated technologies, such as next-generation sequencing (NGS), into routine clinical practice requires compatibility with common tumor biopsy types, such as ...formalin-fixed, paraffin-embedded (FFPE) and fine-needle aspiration specimens, and validation metrics for platforms, controls, and data analysis pipelines. In this study, a two-step PCR enrichment workflow was used to assess 540 known cancer-relevant variants in 16 oncogenes for high-depth sequencing in tumor samples on either mature (Illumina GAIIx) or emerging (Ion Torrent PGM) NGS platforms. The results revealed that the background noise of variant detection was elevated approximately twofold in FFPE compared with cell line DNA. Bioinformatic algorithms were optimized to accommodate this background. Variant calls from 38 residual clinical colorectal cancer FFPE specimens and 10 thyroid fine-needle aspiration specimens were compared across multiple cancer genes, resulting in an accuracy of 96.1% (95% CI, 96.1% to 99.3%) compared with Sanger sequencing, and 99.6% (95% CI, 97.9% to 99.9%) compared with an alternative method with an analytical sensitivity of 1% mutation detection. A total of 45 of 48 samples were concordant between NGS platforms across all matched regions, with the three discordant calls each represented at <10% of reads. Consequently, NGS of targeted oncogenes in real-life tumor specimens using distinct platforms addresses unmet needs for unbiased and highly sensitive mutation detection and can accelerate both basic and clinical cancer research.
In the present work, an attempt has been made to detect carbon dioxide (CO
2
) using an in situ-loaded Bi
2
O
3
–polypyrrole nanocomposite sensor. The polypyrrole (PPy) was prepared by oxidative ...polymerization method from precursor’s monomer pyrrole and aqueous solution of ammonium persulphate. The prepared nanocomposites were characterized by X-ray diffractometry (XRD), scanning electron microscopy (SEM), ultraviolet–visible spectroscopy (UV–Vis), thermogravimetric–differential thermal analysis (TG–DTA) and Raman spectroscopy. Thick films of the synthesized PPy nanocomposites were deposited on glass substrate by screen printing technique. Resistive type gas sensors are extensively used in a number of modern applications such as emission control and energy efficiency. By considering this view, PPy nanocomposite films were prepared for gas sensing application. The XRD analysis revealed amorphous nature. The SEM micrograph showed that nanocomposite particles have irregular morphology and size. The nanocomposites showed a good sensing performance toward CO
2
gas. The highest value of CO
2
gas sensing response was shown by 25 wt% Bi
2
O
3
-loaded PPy nanocomposite sample (S5), which was found to be 0.065 for 50 ppm. Operating temperature for the optimized sample S5 toward CO
2
gas was found to be 363 K. Similarly, the optimized sample S5 showed an excellent stability response against CO
2
gas and atmospheric change for 30 days.
Introduction: Provisional restorative materials require good mechanical properties for long term restorations. Provisional restorative materials are continuously being updated and improved upon for ...these mechanical, biological and esthetic properties. Aim: To determine and compare the surface roughness and the strength of three different commercially available provisional restorative materials at different periods. Materials and Methods: This in-vitro study involved 120 samples of a stainless steel mould fabricated and equally divided. The study was carried out for one year in the Department of Prosthodontics and Crown & Bridge, School of Dental Sciences, Sharda University between June 2020 to June 2021. Materials compared were Dental Products of India (DPI) heat cure acrylic, Protemp 4 and Revotek LC. Each group was further categorised into four groups to measure the flexural strength (after 24 hours, storage in artificial saliva for seven days and 30 days) and surface roughness. A Universal Testing Machine determined the flexural strength and Perthometer for surface roughness. Statistical analysis was done using a one-way Analysis of Variance (ANOVA) test followed by a post-hoc test. Results: In the three types of materials studied, flexural strength after 24 hours, 7 days, and 30 days was statistically significantly higher (p-value=0.001) in DPI than Protemp 4 and Revotek LC whereas, DPI exhibited a decrease in surface roughness when compared to Protemp 4 and Revotek LC (p-value=0.002). Conclusion: Within the limitations of the study it was found that DPI demonstrated a higher flexural strength at 24 hours than Protemp 4 and Revotek LC. After storage in saliva for 7 days and 30 days, there was a decrease in the flexural strength of three commercially available provisional materials. The mean surface roughness of Revotek LC was more than DPI and Protemp 4.
Development of head and neck squamous cell carcinoma (HNSCC) is characterized by accumulation of mutations in several oncogenes and tumor suppressor genes. We have formerly described the mutation ...pattern of HNSCC and described NOTCH signaling pathway alterations. Given the complexity of the HNSCC, here we extend the previous study to understand the overall HNSCC mutation context and to discover additional genetic alterations. We performed high depth targeted exon sequencing of 51 highly actionable cancer-related genes with a high frequency of mutation across many cancer types, including head and neck. DNA from primary tumor tissues and matched normal tissues was analyzed for 37 HNSCC patients. We identified 26 non-synonymous or stop-gained mutations targeting 11 of 51 selected genes. These genes were mutated in 17 out of 37 (46%) studied HNSCC patients. Smokers harbored 3.2-fold more mutations than non-smokers. Importantly, TP53 was mutated in 30%, NOTCH1 in 8% and FGFR3 in 5% of HNSCC. HPV negative patients harbored 4-fold more TP53 mutations than HPV positive patients. These data confirm prior reports of the HNSCC mutational profile. Additionally, we detected mutations in two new genes, CEBPA and FES, which have not been previously reported in HNSCC. These data extend the spectrum of HNSCC mutations and define novel mutation targets in HNSCC carcinogenesis, especially for smokers and HNSCC without HPV infection.
Auxin levels are tightly regulated within the plant cell, and its storage in the isolated cavity of proteins is a measure adopted by cells to maintain the availability of auxin. We report the first ...crystal structure of Wrightia tinctoria 11S globulin (WTG) in complex with Indole-3-acetic acid (IAA), an auxin, at 1.7 Å resolution. WTG hexamers assemble as a result of the stacking interaction between the hydrophobic surfaces of two trimers, leaving space for the binding of charged ligands. The bound auxin is stabilized by non-covalent interactions, contributed by four chains in each cavity. The presence of bound ligand was confirmed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and high-resolution mass spectrometry (HRMS). Here, we hypothesize that the cleavage of globulins by endopeptidases leads to the movement of the hydrophilic loop region from the surface to the periphery, leaving space for the binding of auxin, and promotes hexamer formation. As the process of germination proceeds, there is a change in the pH, which induces the dissociation of the hexamer and the release of auxin. The compact hexameric assembly ensures the long-term, stable storage of the hormone. This suggests a role for globulin as a novel player in auxin homeostasis.
Introduction: Poor mechanical properties are among the main limitations of denture base resin. There has been a continuous attempt to improve the mechanical properties of denture base resins. ...Nanotechnology has evolved health care industry to a large scale and its applications are a boon to modern medicine and dental science. Nanoparticles are nowadays, extensively used in prosthodontics as they are incorporated in Polymethyl Methacrylate denture bases to alter the properties such as impact strength. Aim: To evaluate and compare the effect of titanium dioxide and silicon dioxide nanoparticles on impact strength of two commercially available heat cure acrylic resins. Materials and Methods: The in-vitro study was conducted in Department of Prosthodontics and Crown and Bridge, School of Dental Sciences, Sharda University Faridabad, Haryana, India between April 2019 to May 2021, that involved 120 samples. Materials compared were Dental Products of India (DPI) heat cure acrylic and Trevalon heat cure acrylic. Each group was further categorized into four groups to measure the impact strength i.e, without incorporation of nanoparticles and with incorporation of nanoparticles SiO2 and TiO2 and a combination of both. Samples obtained were tested for impact strength using Izod method. Statistical analysis was done using a one-way Analysis of Variance (ANOVA), Student’s t-test and Post-hoc Bonferroni test. Results: In the two types of materials studied, the mean impact strength of Trevalon was statistically significantly higher (p-value= 0.045) than DPI. After the addition of nanoparticles, i.e; SiO2 and TiO2 the mean impact strength was higher in Trevalon (8.66 kJ/m2 without the addition of nanoparticles, 5.79 kJ/m2 addition of 1% TiO2 nanoparticles, 5.77 kJ/m2 addition of 1% SiO2 nanoparticles and 5.75 kJ/m2 when a 1% combination of both the above was added) than DPI (7.19 kJ/m2 without the addition of nanoparticles, 5.86 kJ/m2 - addition of 1% TiO2 nanoparticles, 5.77 kJ/m2 addition of 1% SiO2 nanoparticles and 5.66 kJ/m2 when a 1% combination of both the above was incorporated). Conclusion: Mean Impact strength of Trevalon was higher than mean impact strength of DPI. Incorporation of TiO2 , SiO2 Nanoparticles or in combination decreases impact strength of both the commercially available heat cure denture base resin with statistically no significant difference.
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