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Abstract Background We recently found that epiplakin 1 (EPPK1) alterations were present in 12% of lung adenocarcinoma (LUAD) cases and were associated with a poor prognosis in early-stage LUAD when combined with other molecular alterations. This study aimed to identify a probable crucial role for EPPK1 in cancer development. Methods EPPK1 mRNA and protein expression was analyzed with clinical variables. Normal bronchial epithelial cell lines were exposed to cigarette smoke for 16 weeks to determine whether EPPK1 protein expression was altered after exposure. Further, we used CRISPR-Cas9 to knock out (KO) EPPK1 in LUAD cell lines and observed how the cancer cells were altered functionally and genetically. Results EPPK1 protein expression was associated with smoking and poor prognosis in early-stage LUAD. Moreover, a consequential mesenchymal-to-epithelial transition was observed, subsequently resulting in diminished cell proliferation and invasion after EPPK1 KO. RNA sequencing revealed that EPPK1 KO induced downregulation of 11 oncogenes, 75 anti-apoptosis, and 22 angiogenesis genes while upregulating 8 tumor suppressors and 12 anti-cell growth genes. We also observed the downregulation of MYC and upregulation of p53 expression at both protein and RNA levels following EPPK1 KO. Gene ontology enrichment analysis of molecular functions highlighted the correlation of EPPK1 with the regulation of mesenchymal cell proliferation, mesenchymal differentiation, angiogenesis, and cell growth after EPPK1 KO. Conclusions Our data suggest that EPPK1 is linked to smoking, epithelial to mesenchymal transition, and the regulation of cancer progression, indicating its potential as a therapeutic target for LUAD.

Abstract Lung adenocarcinoma (LUAD) is the predominant type of lung cancer in the U.S. and exhibits a broad variety of behaviors ranging from indolent to aggressive. Identification of the biological determinants of LUAD behavior at early stages can improve existing diagnostic and treatment strategies. Extracellular matrix (ECM) remodeling and cancer-associated fibroblasts play a crucial role in the regulation of cancer aggressiveness and there is a growing need to investigate their role in the determination of LUAD behavior at early stages. We analyzed tissue samples isolated from patients with LUAD at early stages and used imaging-based biomarkers to predict LUAD behavior. Single-cell RNA sequencing and histological assessment showed that aggressive LUADs are characterized by a decreased number of ADH1B+ CAFs in comparison to indolent tumors. ADH1B+ CAF enrichment is associated with distinct ECM and immune cell signatures in early-stage LUADs. Also, we found a positive correlation between the gene expression of ADH1B+ CAF markers in early-stage LUADs and better survival. We performed TCGA dataset analysis to validate our findings. Identified associations can be used for the development of the predictive model of LUAD aggressiveness and novel therapeutic approaches.

Introduction: Lung cancer is the deadliest cancer in the United States and worldwide, and lung adenocarcinoma (LUAD) is the most prevalent histologic subtype in the United States. LUAD exhibits a wide range of aggressiveness and risk of recurrence, but the biological underpinnings of this behavior are poorly understood. Past studies have focused on the biological characteristics of the tumor itself, but the ability of the immune response to contain tumor growth represents an alternative or complementary hypothesis. Emerging technologies enable us to investigate the spatial distribution of specific cell types within the tumor nest and characterize this immune response. This study aimed to investigate the association between immune cell density within the primary tumor and recurrence-free survival (RFS) in stage I and II LUAD. Methods: This study is a prospective collection with retrospective evaluation. A total of 100 patients with surgically resected LUAD and at least 5-year follow-ups, including 69 stage I and 31 stages II tumors, were enrolled. Multiplexed immunohistochemistry panels for immune markers were used for measurement. Results: Cox regression models adjusted for sex and EGFR mutation status revealed that the risk of recurrence was reduced by 50% for the unit of one interquartile range (IQR) change in the tumoral T-cell (adjusted hazard ratio per IQR increase = 0.50, 95% confidence interval: 0.27–0.93) and decreased by 64% in mast cell density (adjusted hazard ratio per IQR increase = 0.36, confidence interval: 0.15–0.84). The analyses were reported without the type I error correction for the multiple types of immune cell testing. Conclusions: Analysis of the density of immune cells within the tumor and surrounding stroma reveals an association between the density of T-cells and RFS and between mast cells and RFS in early-stage LUAD. This preliminary result is a limited study with a small sample size and a lack of an independent validation set.

Background 18F-fluorodeoxyglucose (FDG) PET/CT is recommended for evaluation of intermediate-risk indeterminate pulmonary nodules (IPNs). While highly sensitive, the specificity of FDG remains suboptimal for differentiating malignant from benign nodules, particularly in areas where fungal lung diseases are prevalent. Thus, a cancer-specific imaging probe is greatly needed. In this study, we tested the hypothesis that a PET radiotracer (S)-4-(3-[18F]-fluoropropyl)-L-glutamic acid (FSPG) improves the diagnostic accuracy of IPNs compared to 18F-FDG PET/CT. Methods This study was conducted at a major academic medical center and an affiliated VA medical center. Twenty-six patients with newly discovered IPNs 7-30mm diameter or newly diagnosed lung cancer completed serial PET/CT scans utilizing 18F-FDG and 18F-FSPG, without intervening treatment of the lesion. The scans were independently reviewed by two dual-trained diagnostic radiology and nuclear medicine physicians. Characteristics evaluated included quantitative SUVmax values of the pulmonary nodules and metastases. Results A total of 17 out of 26 patients had cancer and 9 had benign lesions. 18F-FSPG was negative in 6 of 9 benign lesions compared to 7 of 9 with 18F-FDG. 18F-FSPG and 18F-FDG were positive in 14 of 17 and 12 of 17 malignant lesions, respectively. 18F-FSPG detected brain and intracardiac metastases missed by 18F-FDG PET in one case, while 18F-FDG detected a metastasis to the kidney missed by 18F-FSPG. Conclusion In this pilot study, there was no significant difference in overall diagnostic accuracy between 18F-FSPG and 18F-FDG for the evaluation of IPNs and staging of lung cancer. Additional studies will be needed to determine the clinical utility of this tracer in the management of IPNs and lung cancer.

Introduction: Blood-based next-generation sequencing assays of circulating tumor DNA (ctDNA) have the ability to detect tumor-associated mutations in patients with SCLC. We sought to characterize the relationship between ctDNA mean variant allele frequency (VAF) and radiographic total-body tumor volume (TV) in patients with SCLC. Methods: We identified matched blood draws and computed tomography (CT) or positron emission tomography (PET) scans within a prospective SCLC blood banking cohort. We sequenced plasma using our previously developed 14-gene SCLC-specific ctDNA assay. Three-dimensional TV was determined from PET and CT scans using MIM software and reviewed by radiation oncologists. Univariate association and multivariate regression analyses were performed to evaluate the association between mean VAF and total-body TV. Results: We analyzed 75 matched blood draws and CT or PET scans from 25 unique patients with SCLC. Univariate analysis revealed a positive association between mean VAF and total-body TV (Spearman’s ρ = 0.292, p < 0.01), and when considering only treatment-naive and pretreatment patients (n = 11), there was an increase in the magnitude of association (ρ = 0.618, p = 0.048). The relationship remained significant when adjusting for treatment status and bone metastases (p = 0.046). In the subgroup of patients with TP53 variants, univariate analysis revealed a significant association (ρ = 0.762, p = 0.037) only when considering treatment-naive and pretreatment patients (n = 8). Conclusions: We observed a positive association between mean VAF and total-body TV in patients with SCLC, suggesting mean VAF may represent a dynamic biomarker of tumor burden that could be followed to monitor disease status.

Animal miRNAs commonly mediate mRNA degradation and/or translational repression by binding to their target mRNAs. Key factors for miRNA-mediated mRNA degradation are the components of the miRNA effector complex (AGO1 and GW182) and the general mRNA degradation machinery (deadenylation and decapping enzymes). The CCR4-NOT1 complex required for the deadenylation of target mRNAs is directly recruited to the miRNA effector complex. However, it is unclear whether the following decapping step is only a consequence of deadenylation occurring independent of the miRNA effector complex or e.g. decapping activators can get recruited to the miRNA effector complex. In this study we performed split-affinity purifications in Drosophila cells and provide evidence for the interaction of the decapping activator HPat with the miRNA effector complex. Furthermore, in knockdown analysis of various mRNA degradation factors we demonstrate the importance of NOT1 for this interaction. This suggests that deadenylation and/or the recruitment of NOT1 protein precedes the association of HPat with the miRNA effector complex. Since HPat couples deadenylation and decapping, the recruitment of HPat to the miRNA effector complex provides a mechanism to commit the mRNA target for degradation.

In recent years, unmanned aerial vehicles (UAVs) have become increasingly important as a tool for quickly collecting high-resolution (spatial and spectral) imagery of the Earth's surface. The final products are highly dependent on the choice of values for various parameters in flight planning, the type of sensors, and the processing of the data. In this paper ground control points (GCPs) were first measured using the Global Navigation Satellite System (GNSS) Real-Time Kinematic (RTK) method, and then due to the low height accuracy of the GNSS RTK method all points were measured using a detailed leveling method. This study aims to provide a basic assessment of quality, including four main aspects: (1) the difference between an RGB sensor and a five-band multispectral sensor on accuracy and the amount of data, (2) the impact of the number of GCPs on the accuracy of the final products, (3) the impact of different altitudes and cross flight strips, and (4) the accuracy analysis of multi-altitude models. The results suggest that the type of sensor, flight configuration, and GCP setup strongly affect the quality and quantity of the final product data while creating a multi-altitude model does not result in the expected quality of data. With its unique combination of sensors and parameters, the results and recommendations presented in this paper can assist professionals and researchers in their future work.

Extreme precipitation events, which are common natural hazards, are expected to increase in frequency due to global warming, leading to various types of floods, including pluvial floods. In this study, we investigated the probabilities of maximum 3-day precipitation amount (Rx3day) occurrences during spring in the Vojvodina region, covering both past (1971–2019) and future (2020–2100) periods. We utilized an ensemble of eight downscaled, bias-corrected regional climate models from the EURO-CORDEX project database, selecting the RCP8.5 scenario to examine future Rx3day amounts. The probabilities of occurrences of Rx3day were modeled using the GEV distribution, while the number of events where Rx3day in spring exceeds specific thresholds was modeled using the Poisson distribution. The results indicate that Rx3day with a ten-year return period during the spring months is expected to increase by 19% to 33%. Additionally, the probabilities of having more than one event where Rx3day exceeds thresholds are projected to rise by 105.6% to 200.0% in the future compared to the historical period. The analysis comparing the design values of Rx3day with future projections for the period 2020–2100 revealed that 51 drainage systems are likely to function without difficulties under future climate conditions. However, for the remaining 235 drainage systems, an increased risk of pluvial flooding was identified, as their design precipitation amounts are lower than the future projections. This study reveals that analyzing extreme rainfall events in the context of climate change yields crucial information that facilitates effective planning and policy making in water management, particularly flood protection.

With the continuous trend of urbanization, increase in industrial capacities, and expansion of agricultural areas, there is also a rise in the amount of wastewater. One of the effective and economical solutions for wastewater treatment has proven to be Constructed Wetlands (CWs). Defining the locations where CWs can be built is not an easy task and there are several criteria that need to be considered. The Geographical Information Systems (GIS) and Multi-Criteria Decision Analysis—Analytic Hierarchy Process (AHP) are combined to select CW locations. AHP is one of the most commonly used methods in many environmental decision making problems, involving various conflicting criteria. In this case, conflicts arise between the evaluation of criteria that influence the selection of CW locations. The evaluation of selected criteria and sub-criteria resulted in a suitability map indicating that the first class represents 44%, the second class 37%, and the third class 16% of the total area. The fourth and fifth classes represent 3% of the total area. The criteria with the highest significance are land use, floodplains and distance of the location from populated places. This study has important implications for sustainable wastewater management in Serbia and provides guidelines for selecting locations for CWs.

Liquid-level control in tanks is widely used in various industrial sectors. Due to problems arising from the case that the liquid level is above or below the recommended, the process of its control is of great significance. The proportional integral differential (PID) controller is one of the most widely used controllers in applications that require accuracy and optimal automatic control. In this paper, single and coupled ship fuel tank systems are discussed. In addition, mathematical models and their linearization using two different approaches are presented. The two control approaches are used to maintain the specified liquid level in tanks. The first is based on the design of standard PID controllers, with the application of the Ziegler–Nichols, Takahashi tuning, and Auto-tuning methods. The second approach, proposed by the authors, is based on the application of the Dahlin algorithm. The simple modification of the Dahlin controller, which can avoid the “ringing” of the control signal, is suggested in the paper. Moreover, the lower limit of the decrease in the desired time constant, which ensures the desired behavior of the system, is determined. The comparisons provided for different performance indexes show the advantages of the modified Dahlin algorithm approach compared to the standard PID controllers. Furthermore, the comparison with the literature-known approaches is realized.