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Abstract Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal lung disease with a poor prognosis. Current/available clinical prediction tools have limited sensitivity and accuracy when evaluating clinical outcomes of IPF. Research has shown that focal adhesion kinase (FAK), produced by the protein tyrosine kinase 2 (PTK2) gene, is crucial in IPF development. FAK activation is a characteristic of lesional fibroblasts; Thus, FAK may be a valuable therapeutic target or prognostic biomarker for IPF. This study aimed to create a gene signature based on PTK2-associated genes and microarray data from blood cells to predict disease prognosis in patients with IPF. PTK2 levels were found to be higher in lung tissues of IPF patients compared to healthy controls, and PTK2 inhibitor Defactinib was found to reduce TGFβ-induced FAK activation and increase α-smooth muscle actin. Although the blood PTK2 levels were higher in IPF patients, blood PTK level alone could not predict IPF prognosis. From 196 PTK2-associated genes, 11 genes were prioritized to create a gene signature (PTK2 molecular signature) and a risk score system using univariate and multivariate Cox regression analysis. Patients were divided into high-risk and low-risk groups using PTK2 molecular signature. Patients in the high-risk group experienced decreased survival rates compared to patients in the low-risk group across all discovery and validation cohorts. Further functional enrichment and immune cell proportion analyses revealed that the PTK2 molecular signature strongly reflected the activation levels of immune pathways and immune cells. These findings suggested that PTK2 is a molecular target of IPF and the PTK2 molecular signature is an effective IPF prognostic biomarker.

Background Cellular immunotherapies using autologous tumor-infiltrating lymphocytes (TIL) can induce durable regression of epithelial cancers in selected patients with treatment-refractory metastatic disease. As the genetic engineering of T cells with tumor-reactive T-cell receptors (TCRs) comes to the forefront of clinical investigation, the rapid, scalable, and cost-effective detection of patient-specific neoantigen-reactive TIL remains a top priority.Methods We analyzed the single-cell transcriptomic states of 31 neoantigen-specific T-cell clonotypes to identify cell surface dysfunction markers that best identified the metastatic transcriptional states enriched with antitumor TIL. We developed an efficient method to capture neoantigen-reactive TCRs directly from resected human tumors based on cell surface co-expression of CD39, programmed cell death protein-1, and TIGIT dysfunction markers (CD8+ TILTP).Results TILTP TCR isolation achieved a high degree of correlation with single-cell transcriptomic signatures that identify neoantigen-reactive TCRs, making it a cost-effective strategy using widely available resources. Reconstruction of additional TILTP TCRs from tumors identified known and novel antitumor TCRs, showing that at least 39.5% of TILTP TCRs are neoantigen-reactive or tumor-reactive. Despite their substantial enrichment for neoantigen-reactive TCR clonotypes, clonal dynamics of 24 unique antitumor TILTP clonotypes from four patients indicated that most in vitro expanded TILTP populations failed to demonstrate neoantigen reactivity, either by loss of neoantigen-reactive clones during TIL expansion, or through functional impairment during cognate neoantigen recognition.Conclusions While direct usage of in vitro-expanded CD8+ TILTP as a source for cellular therapy might be precluded by profound TIL dysfunction, isolating TILTP represents a streamlined effective approach to rapidly identify neoantigen-reactive TCRs to design engineered cellular immunotherapies against cancer.

Abstract Background Vitamin C is one of the most important micronutrient required for various physiological roles in the human system. Evidences suggest that there is an inadequate status of vitamin C in diabetes mellitus. The objectives of this study is to understand the prevalence of vitamin C deficiency in established type II diabetes mellitus patients and to study the correlation between various variables of diabetes mellitus with serum vitamin C levels. Methods A prospective cross-sectional study to assess the prevalence of vitamin C deficiency was carried out in diabetes patients. Fasting blood sugar levels, glycated hemoglobin, serum malondialdehyde levels, and lipid profile levels were correlated with serum vitamin C levels. Results The prevalence rate of hypovitaminosis C is found to be 55.13% among the enrolled patients. There is a significant increase in the systolic blood pressure levels in patients with inadequate as well as deficient vitamin C levels (p < 0.05). Inverse relationship exists between fasting blood sugar and vitamin C levels (p < 0.001). Similarly total cholesterol levels were also inversely related to the vitamin C levels (p = 0.0031). Body mass index, glycated hemoglobin, and fasting blood sugar levels are important predictors of vitamin C deficiencies. Conclusion Vitamin C deficiency is well established among diabetes mellitus patients. Deficiency of vitamin C levels has an impact on the serum malondialdehyde levels suggesting increased oxidative stress. The higher oxidative stress would have led to increase in glycated hemoglobin. Further research must be carried out to understand the beneficial effects of vitamin C supplementation in diabetes mellitus.

A simple, selective and sensitive stability indicating high-performance thin-layer chromatographic method was developed and validated for estimation of cyclobenzaprine hydrochloride (CBP) in bulk drug and commercial tablets according to ICH guidelines. A precoated silica gel 60F254 HPTLC plates were used for chromatographic separation using mobile phase toluene: ethyl acetate: methanol: glacial acetic acid in the ratio 4:2:3.5:0.5 v/v/v/v. A TLC scanner was set at detection wavelength 290 nm for densitometric analysis of analyte on absorbance mode. The degradants were resolved satisfactorily in a mixture of stressed sample having Rf values 0.48 ± 0.05, 0.52 ± 0.05, 0.86 ± 0.05, 0.66 ± 0.05 and 0.27 ± 0.05. The calibration curve of drug was found linear in the concentration range 200–1000 ng/band. The (r2 > 0.999) correlation coefficient value indicated good correlation between the analyte concentrations and peak areas. The repeatability and intermediate precision study revealed the % RSD value less than 2.0 and was found to be satisfactory. The accuracy of developed method was ascertained by performing the recovery study using standard addition method and expressed by percent recovery (100.32%) which was found satisfactory. CBP was subjected to various stress conditions as per International Conference on Harmonization (ICH) guidelines. The drug showed significant degradation during hydrolytic and oxidative stress condition. CBP remained stable in thermal and photolytic stress testing. The validated chromatographic method was further utilized to isolate the alkaline degradation product using preparative HPTLC technique and extensive FT-IR, ESI-MS/TOF studies were performed to ascertain the structure of degradant.

A sensitive stability indicating high-performance thin-layer chromatographic method was developed and validated for quantitative determination of Acyclovir in tablets. Chromatographic separation was performed on a precoated silica gel 60F254 HPTLC plates using Toluene: n-Butanol: Methanol: Water (5.0:3.0:1.0:1.0 v/v/v/v) as a mobile phase. A TLC scanner set at 259 nm was used for direct evaluation of the chromatograms in reflectance/absorbance mode. Acyclovir and degradant were satisfactorily resolved with Rf values of 0.62 ± 0.05, 0.78 ± 0.05, respectively. Calibration curve was a polynomial in the concentration range of 200–1000 ng/band. The high correlation coefficient (r2 > 0.9991) values indicated clear correlations between the investigated compound concentrations and their peak areas within the test ranges. The method was validated according to ICH guidelines. The repeatability and intermediate precision, expressed by the RSD, were less than 2.0%. The accuracy and validity of the method were further ascertained by performing recovery studies via a standard addition method. The accuracy of the method expressed as percent recovery was satisfactory (99.85%). The drug was subjected to the International Conference on Harmonization (ICH)-prescribed hydrolytic, oxidative, photolytic and thermal stress conditions. The method was validated according to ICH guidelines. The drug showed instability in alkaline and oxide while it remained stable in heat and UV radiation conditions. The proposed HPTLC method was utilized to investigate the alkaline degradation of Acyclovir (ACY). The performance of the method was validated according to the present ICH guidelines for specificity, limit of detection, limit of quantification, linearity, accuracy, precision, ruggedness and robustness.

Clathrin-mediated endocytosis (CME) involves nanoscale bending and inward budding of the plasma membrane, by which cells regulate both the distribution of membrane proteins and the entry of extracellular species. Extensive studies have shown that CME proteins actively modulate the plasma membrane curvature. However, the reciprocal regulation of how the plasma membrane curvature affects the activities of endocytic proteins is much less explored, despite studies suggesting that membrane curvature itself can trigger biochemical reactions. This gap in our understanding is largely due to technical challenges in precisely controlling the membrane curvature in live cells. In this work, we use patterned nanostructures to generate well-defined membrane curvatures ranging from +50 nm to -500 nm radius of curvature. We find that the positively curved membranes are CME hotspots, and that key CME proteins, clathrin and dynamin, show a strong preference towards positive membrane curvatures with a radius

BACKGROUND. Therapeutic vaccinations against cancer have mainly targeted differentiation antigens, cancer-testis antigens, and overexpressed antigens and have thus far resulted in little clinical benefit. Studies conducted by multiple groups have demonstrated that T cells recognizing neoantigens are present in most cancers and offer a specific and highly immunogenic target for personalized vaccination. METHODS. We recently developed a process using tumor-infiltrating lymphocytes to identify the specific immunogenic mutations expressed in patients' tumors. Here, validated, defined neoantigens, predicted neoepitopes, and mutations of driver genes were concatenated into a single mRNA construct to vaccinate patients with metastatic gastrointestinal cancer. RESULTS. The vaccine was safe and elicited mutation-specific T cell responses against predicted neoepitopes not detected before vaccination. Furthermore, we were able to isolate and verify T cell receptors targeting [KRAS.sup.G12D] mutation. We observed no objective clinical responses in the 4 patients treated in this trial. CONCLUSION. This vaccine was safe, and potential future combination of such vaccines with checkpoint inhibitors or adoptive T cell therapy should be evaluated for possible clinical benefit in patients with common epithelial cancers. TRIAL REGISTRATION. Phase I/II protocol (NCT03480152) was approved by the IRB committee of the NIH and the FDA. FUNDING. Center for Clinical Research, NCI, NIH., Introduction Protective vaccination against infectious diseases has proven to be one of the most effective health measures in medicine; however, therapeutic vaccination against established diseases such as persistent infections and [...]

The biodegradability and lightweight qualities of magnesium make it an appealing material for usage in aerospace and automotive applications and orthopedics. Magnesium is susceptible to corrosion, which restricts its use. Magnesium should be protected against corrosion in order to be used to its maximal potential. Corrosion inhibitors are one of the most well-liked and affordable techniques to stop corrosion in magnesium and its alloys. Chemicals, which are frequently used as corrosion inhibitors include biotoxic ones, which raise serious toxicity issues. There is a great deal of interest in creating a green corrosion inhibitor, which is non-toxic and safe for the environment due to the toxicity of the current inhibitors. This paper discusses the common green corrosion inhibitors. The application of ecologically friendly corrosion inhibitors for magnesium and its alloy is still being pursued by a relatively small number of scientists. The research utilizing green corrosion inhibitors for magnesium alloy has been reviewed in detail. In addition, the performances are tabulated and analyzed. [ABSTRACT FROM AUTHOR]