Project description:Arabidopsis plants (Col-0) were exposed to heat stress (40oC) or microwave irradiation (23W) for 1h. Shoots were sampled and transcripts significantly up- or down-regulated by heat stress or microwave irradiation were investigated.
Project description:Reports that low-intensity microwave radiation can induce heat-shock reporter gene expression in the nematode, Caenorhabditis elegans, have recently been reinterpreted as a subtle thermal effect caused by very slight heating. This study used a microwave exposure system (1.0 GHz, 0.5 W power input; SAR 0.9-3 mW kg-1 for 6-well plates) that minimises the temperature differential between sham and exposed conditions to ≤ 0.1°C. Comparable measurement and simulation studies of SAR distribution within this exposure system are presented. We compared 5 Affymetrix gene-arrays of pooled triplicate RNA populations from sham-exposed L4/adult worms against 5 gene-arrays of pooled RNA from microwave-exposed worms (taken from the same source population in each run). Few genes showed consistent expression changes across all 5 comparisons, and all such expression changes appeared modest after applying standard normalisation procedures (≤ 30% up- or down-regulated). The number of statistically significant differences in gene expression (846) was less than the false-positive rate expected by chance (1131). As one example, an apparent up-regulation of the vit-3 vitellogenin gene by microwave exposure was not mirrored by similar changes affecting the other co-regulated members of the same vit gene family. We conclude that the pattern of gene expression in L4/adult C elegans is not substantially perturbed by low-intensity microwave radiation, and that the minor changes observed in this study may well be explicable as false positives. As a check on the sensitivity of the Affymetrix gene-arrays used, we also compared RNA samples from N2 worms subjected to a sub-heat-shock treatment (28ºC) against controls kept at 26 ºC (but using only 2 gene arrays per condition). After similar normalisation, many more genes (3712) showed substantial expression changes (i.e. > 2-fold at p < 0.05), including a group of six heat-shock genes which were strongly but unexpectedly down-regulated (by > 10-fold). However, further replication and confirmation by real-time RT-PCR would be needed to establish how many of these changes might also be false positives. Experimenter name: Adam Dawe Experimenter phone: +27 21 959 2364 adam@sanbi.ac.za Experimenter institute: South African National Bioinformatics Institute Experimenter address: University of Western Cape, Old Chemistry Building, University of Western Cape, Modderdam Road, Bellville 7530, Capetown Experimenter zip/postal_code: 7530 Experimenter country: South Africa Keywords: Microwave radiation, gene expression, gene arrays, Caenorhabditis elegans
Project description:Reports that low-intensity microwave radiation can induce heat-shock reporter gene expression in the nematode, Caenorhabditis elegans, have recently been reinterpreted as a subtle thermal effect caused by very slight heating. This study used a microwave exposure system (1.0 GHz, 0.5 W power input; SAR 0.9-3 mW kg-1 for 6-well plates) that minimises the temperature differential between sham and exposed conditions to ≤ 0.1°C. Comparable measurement and simulation studies of SAR distribution within this exposure system are presented. We compared 5 Affymetrix gene-arrays of pooled triplicate RNA populations from sham-exposed L4/adult worms against 5 gene-arrays of pooled RNA from microwave-exposed worms (taken from the same source population in each run). Few genes showed consistent expression changes across all 5 comparisons, and all such expression changes appeared modest after applying standard normalisation procedures (≤ 30% up- or down-regulated). The number of statistically significant differences in gene expression (846) was less than the false-positive rate expected by chance (1131). As one example, an apparent up-regulation of the vit-3 vitellogenin gene by microwave exposure was not mirrored by similar changes affecting the other co-regulated members of the same vit gene family. We conclude that the pattern of gene expression in L4/adult C elegans is not substantially perturbed by low-intensity microwave radiation, and that the minor changes observed in this study may well be explicable as false positives. As a check on the sensitivity of the Affymetrix gene-arrays used, we also compared RNA samples from N2 worms subjected to a sub-heat-shock treatment (28ºC) against controls kept at 26 ºC (but using only 2 gene arrays per condition). After similar normalisation, many more genes (3712) showed substantial expression changes (i.e. > 2-fold at p < 0.05), including a group of six heat-shock genes which were strongly but unexpectedly down-regulated (by > 10-fold). However, further replication and confirmation by real-time RT-PCR would be needed to establish how many of these changes might also be false positives. Experimenter name: Adam Dawe; Experimenter phone: +27 21 959 2364; adam@sanbi.ac.za; Experimenter institute: South African National Bioinformatics Institute; Experimenter address: University of Western Cape, Old Chemistry Building, University of Western Cape, Modderdam Road, Bellville 7530, Capetown; Experimenter zip/postal_code: 7530; Experimenter country: South Africa Experiment Overall Design: 14 samples were used in this experiment
Project description:High temperature stress results in yield loss and alterations to seed composition during seed filling in oilseed rape (Brassica napus). However, the mechanism underlying this heat response is poorly understood. In this study, we employed a microarray analysis with silique walls and seeds from the developing siliques (20 days after flowering) of Brassica napus that had undergone heat stress. Two-condition experiment, control vs heat stress, 2 time points
Project description:We introduce a new reagent – Trimethylacetic anhydride (TMA) for highly efficient histone derivatization, which is facilitated by microwave irradiation to reduce sample preparation duration. The derivatization of unmodified NH2- groups at histone lysine residues is necessary prior LC-MS analysis using bottom-up approach. Our methodology shows high labelling efficiency and demonstrates its utilization for histone posttranslational modification abundance monitoring. Therefore, to be an alternative to conventionally used methods.
Project description:Terahertz (THz) is an electromagnetic wave with a spectrum between microwave and infrared. Due to its unique characteristics, THZ has great application prospects in both medical imaging and biomedicine. However, its effects on the angiogenesis capacity of human umbilical vein endothelial cells (HUVECs) are still unclear. RNA sequencing indicated that the expression of genes related to cytoskeleton increased significantly after THz irradiation.
Project description:Insufficient microwave ablation (IMWA) is linked to aggressive hepatocellular carcinoma (HCC) progression. An increase in lactate level after sublethal heat stress (HS) has been confirmed in HCC. However, the role of lactate-related histone lactylation in the progression of HCC underwent sublethal HS remains unclear. Here, we found that the metastatic potential of HCC is enhanced in a lactate-dependent manner after IMWA. Meanwhile, sublethal HS triggers an upregulation of H3K18la modification, as validated in a cell-derived xenograft mouse model and human HCC samples. By employing an integration analysis of proteomic and transcriptomic profiling, we revealed that HCC cells exhibit an upregulation of intracellular iron ion homeostasis and develop resistance to platinum-based drugs after exposure to sublethal HS. We subsequently integrated proteomic and transcriptomic data with H3K18la-specific chromatin immunoprecipitation (ChIP) sequencing to identify candidate genes involved in sublethal heat treatment-induced HCC cell metastasis. Mechanically, the upregulation of H3K18la modification enhances the transcriptional activity of NFS1, a key player in iron-sulfur clusters biosynthesis, thereby reducing the susceptibility of HCC to ferroptosis post IMWA. Knocking down NFS1 diminished the metastatic potential of sublethal heat-treated HCC cell. Additionally, the deficiency of NFS1 exhibited a synergistic effect with oxaliplatin, leading to a significant inhibition of the metastatic capability of HCC cells both in vitro and in vivo, regardless of HS treatment. In conclusion, our study reveals the oncogenic role of Histone lactylation in HCC after IMVA, we also bridge histone lactylation with ferroptosis, which provides novel therapeutic targets for HCC following microwave ablation, particularly when combined with oxaliplatin-based chemotherapy.
Project description:Terahertz (THz) is an electromagnetic wave with a spectrum between microwave and infrared. Due to its unique characteristics, THZ has great application prospects in both medical imaging and biomedicine. however, its effects on the angiogenesis capacity of human umbilical vein endothelial cells (HUVECs) are still unclear. The assay for transposase-accessible chromatin (ATAC) sequencing indicated that the expression of genes related to cytoskeletal increased significantly after THz irradiation.
Project description:High temperature stress results in yield loss and alterations to seed composition during seed filling in oilseed rape (Brassica napus). However, the mechanism underlying this heat response is poorly understood. In this study, we employed a microarray analysis with silique walls and seeds from the developing siliques (20 days after flowering) of Brassica napus that had undergone heat stress.