Molecular Profiling for Supernatants and Matched Cell Pellets of Pleural Effusions in Non-Small-Cell Lung Cancer.

Pleural effusion (PE) is commonly observed in advanced lung cancer patients. Cell-free total nucleic acid (cfTNA) isolated from cancer patients' plasma has allowed noninvasive tumor genome analyses; however, there are limited studies of detection and characterization of cfTNA in PE. Herein, we included 47 advanced non-small-cell lung cancer patients with PE, who had lung cancer driver mutations tested on tumor tissue specimens either at diagnosis or during disease progression. The supernatant and cell pellet of each PE were evaluated for molecular profiles in parallel on an Ion Torrent next-generation sequencing platform. Somatic mutations were detected in 89.1% supernatant cfTNA, but in only 54.3% of cell pellets. The overall concordance rate between supernatants and formalin-fixed, paraffin-embedded cell pellets at the mutation level was 53.3%. By contrast, 41.7% and 5.0% of somatic alterations were detected in supernatants and cell pellets, respectively. Furthermore, joint analysis of supernatants and cell pellets from PE showed a high concordance (88.3%) of variant detection with their respective tumor tissue specimens. Low-frequency T790M mutations in three cases (0.29%, 0.41%, and 1.56%) were detected in supernatants but not in the matched cell pellets or tumor tissues. In conclusion, pleural effusion-derived cfTNA can effectively be used in clinical practice for molecular analysis by next-generation sequencing, even in cases where corresponding cell pellets or tumor tissues yield insufficient material.

Transcript-Level Analysis in Combination with Real-Time PCR Elucidates Heat Adaptation Mechanism of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) Larvae.

Tribolium castaneum (Herbst) ranks as one of the most prevalent insects in food processing and storage facilities worldwide. Heat treatment has been revisited to disinfest food processing and storage facilities due to increasingly strict regulation on chemicals. The effect of acclimation of T. castaneum larvae to sublethal high temperatures of 36 and 42℃ for 10 h on their heat adaptation was investigated, and transcript-level analysis combinating with real-time PCR (RT-qPCR) was applied for elucidating the heat adaptation mechanism of T. castaneum larvae. Short-term sublethal high temperature acclimation could greatly enhance the thermal adaptability in T. castaneum larvae. In total, 575, 875, and 1017 differentially expressed genes (DEGs) were, respectively, determined in comparisons between the 28 and 36℃ treatments, the 28 and 42℃ treatments, and the 36 and 42℃ treatments. Fifty-three and 96 genes were commonly up- and down-regulated in both the 36 and 42℃ treatments relative to 28℃, respectively. The results of RT-qPCR analysis further confirmed the RNA-seq analysis. The current results are in favor of enhancing the insecticidal effectiveness of extreme high temperature treatment and elucidating the heat adaptation mechanism in T. castaneum larvae.

The effect of acclimation on heat tolerance of Lasioderma serricorne (Fabricius) (Coleoptera: Anobiidae).

Lasioderma serricorne (Fabricius) (Coleoptera: Anobiidae) is an important economic insect of various stored products worldwide. With the restricted use of methyl bromide and the increasing negative effects of repeated use of phosphine fumigation, heat treatment becomes the main approach to control stored-product insects. We investigated effect of the acclimation to sublethal high temperature of 36°C and 42°C for different times on heat tolerance of L. serricorne adults, pupae, larvae, and eggs to lethal high temperature of 50°C in laboratory. The acclimation of L. serricorne pupae, larvae, and eggs, but not adults, to 36°C improved their survival when exposed to lethal high temperature of 50°C. The acclimation of all life stages of L. serricorne to 42°C significantly improved their survival when exposed to 50°C. For eggs, larvae, and adults, the protective effectiveness of acclimation to 42°C was much more profound than that of to 36°C. LT50 and LT90 of all life stages increased with increasing acclimation time to 42°C in general. The LT50 of L. serricorne adults, pupae, larvae, and eggs acclimation to 42°C for 20h were 2.2, 2.2, 3.4, and 4.8 times higher than that of insects without acclimation, respectively. The results suggest that acclimation to sublethal high temperatures can significantly improve the heat tolerance of L. serricorne by the means of increasing their survival when confronting lethal high temperatures. In the face of global warming, the protective effects induced by acclimation to sublethal high temperatures should be fully considered when design or apply heat treatment to control L. serricorne.