Transcriptome analysis of Bacillus subtilis in response to air plasma treatment
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ABSTRACT: In this study, we present a follow up investigation on the effects of argon plasma on vegetative growing Bacillus subtilis 168 (see GSE27113). A growth chamber system suitable for low temperature gas plasma treatment of bacteria in liquid medium enabled a first glimpse on the complex cellular response. In order to gain further knowledge, a second kind of plasma treatment was applied and combined proteomic and transcriptomic analyses were used to investigate the specific stress response of B. subtilis cells to treatment with not only argon but also air plasma. Besides similar cellular responses, due to both argon and air plasma treatment, a variety of different responses such as growth retardation and morphological changes were observed. Furthermore, we analyzed the specific response to DNA damages and oxidative stress. The biological findings such as oxidative stress responses were supported by the detection of reactive plasma species by OES and FTIR measurements.
ORGANISM(S): Bacillus subtilis subsp. subtilis str. 168 Bacillus subtilis
Project description:In this study, we present a follow up investigation on the effects of argon plasma on vegetative growing Bacillus subtilis 168 (see GSE27113). A growth chamber system suitable for low temperature gas plasma treatment of bacteria in liquid medium enabled a first glimpse on the complex cellular response. In order to gain further knowledge, a second kind of plasma treatment was applied and combined proteomic and transcriptomic analyses were used to investigate the specific stress response of B. subtilis cells to treatment with not only argon but also air plasma. Besides similar cellular responses, due to both argon and air plasma treatment, a variety of different responses such as growth retardation and morphological changes were observed. Furthermore, we analyzed the specific response to DNA damages and oxidative stress. The biological findings such as oxidative stress responses were supported by the detection of reactive plasma species by OES and FTIR measurements. Cells were grown in LB medium. At OD540 of 0.5, air plasma treatment was set for 15 min with a discharge power of 7.4 W. Samples for mircroarray analysis were taken 5 min after the 15 min plasma treatment. Microarray hybridizations were performed with RNA from three biological replicates. The individual samples were labeled with Cy5; a reference pool containing equal amounts of RNA from all samples was labeled with Cy3.
Project description:In this study, we introduce for the first time a growth chamber system suitable for physical plasma treatment of bacteria in liquid medium. Bacillus subtilis 168 cells were treated with argon plasma in order to investigate their specific stress response usong a proteomic and transcriptomic approach. The treatment with three different discharge voltages revealed not only growth differences, but also clear cellular stress responses. B. subtilis faces severe cell wall stress, which was made visible alsoelectron microscopy, DNA damages and oxidative stress. The biological findings could be supported by the reactive plasma species, found by plasma diagnostics, i.e. optical emission spectroscopy (OES) and Fourier transformed infrared spectroscopy (FTIR).
Project description:In this study, we introduce for the first time a growth chamber system suitable for physical plasma treatment of bacteria in liquid medium. Bacillus subtilis 168 cells were treated with argon plasma in order to investigate their specific stress response usong a proteomic and transcriptomic approach. The treatment with three different discharge voltages revealed not only growth differences, but also clear cellular stress responses. B. subtilis faces severe cell wall stress, which was made visible alsoelectron microscopy, DNA damages and oxidative stress. The biological findings could be supported by the reactive plasma species, found by plasma diagnostics, i.e. optical emission spectroscopy (OES) and Fourier transformed infrared spectroscopy (FTIR). Cells were grown in LB medium. At OD540 of 0.5, argon plasma treatment was set for 15 min with a discharge power of 5 W. Samples for mircroarray analysis were taken 5 min after the 15 min plasma treatment. Microarray hybridizations were performed with RNA from three biological replicates. The individual samples were labeled with Cy5; a reference pool containing equal amounts of RNA from all samples was labeled with Cy3.
Project description:Treatment of tumor progression and metastasis continues to be of major importance in the field of cancer. It is reported that cancer cells often show a pronounced sensitivity towards oxidative stress. Cold plasma offers the ability to deliver a delicate mix of reactive oxygen and nitrogen species directly into cells or tissues. Using a well-described argon plasma jet, we investigated the biological responses of plasma on tumor cell death, cell migration, and expression of adhesion-associated genes as well as cytoskeletal modifications. Using the human melanoma cancer cell line SK-Mel-147 we were able to show that plasma induced only little apoptosis but had profound effects on tumor cell motility. Plasma treatment of cells was associated with an inhibition of migration and disorganization of the actin cytoskeleton which were mediated through multiple signaling pathways, as transcriptome-wide gene analysis suggested. Specifically, changes in cell adhesion were regulated by differential expression of cell junction and cell-matrix proteins. These results provide evidence that plasma may be able to disturb the migration and adhesion in metastatic SK-Mel-147 cells. Microarrays were used to analyze and investigate the biological effects of cold plasma on human melanoma cell line SK-Mel-147. Using an argon plasma jet kinpen, regulated transcripts were analyzed and further described in Schmidt et al. (2015): M-bM-^@M-^\Human melanoma cell migration and adhesion is decreased by cold plasma treatmentM-bM-^@M-^]. A study using total RNA recovered from human SK-Mel-147, treated with cold plasma as well as H2O2-treated and untreated SK-Mel-147 controls.
Project description:Treatment of tumor progression and metastasis continues to be of major importance in the field of cancer. It is reported that cancer cells often show a pronounced sensitivity towards oxidative stress. Cold plasma offers the ability to deliver a delicate mix of reactive oxygen and nitrogen species directly into cells or tissues. Using a well-described argon plasma jet, we investigated the biological responses of plasma on tumor cell death, cell migration, and expression of adhesion-associated genes as well as cytoskeletal modifications. Using the human melanoma cancer cell line SK-Mel-147 we were able to show that plasma induced only little apoptosis but had profound effects on tumor cell motility. Plasma treatment of cells was associated with an inhibition of migration and disorganization of the actin cytoskeleton which were mediated through multiple signaling pathways, as transcriptome-wide gene analysis suggested. Specifically, changes in cell adhesion were regulated by differential expression of cell junction and cell-matrix proteins. These results provide evidence that plasma may be able to disturb the migration and adhesion in metastatic SK-Mel-147 cells. Microarrays were used to analyze and investigate the biological effects of cold plasma on human melanoma cell line SK-Mel-147. Using an argon plasma jet kinpen, regulated transcripts were analyzed and further described in Schmidt et al. (2015): “Human melanoma cell migration and adhesion is decreased by cold plasma treatment”.
Project description:Oxidative stress illustrates an imbalance between radical formation and removal. Frequent redox stress is critically involved in a variety of human pathologies including cancer, psoriasis, and chronic wounds. However, reactive species pursue a dual role being involved in signaling on the one hand and oxidative damage on the other. Using a HaCaT keratinocyte cell culture model, we here aimed at investigating the cellular and transcriptional response to periodic, low dose oxidative challenge over three months. Chronic redox stress was generated by frequent incubation with cold physical plasma treated cell culture medium. Using mRNA microarray technology, we found both acute ROS stress responses as well as numerous adaptions on the transcriptional level and over several weeks of redox challenge. This included an altered expression of 260 genes that function in inflammation and redox homeostasis, such as, signaling molecules, cytokines, and anti-oxidant enzymes. Apoptotic signaling was affected to a minor extend, especially in p53 down-stream targets. Strikingly, the anti-apoptotic heat shock protein HSP27 was strongly upregulated. These results suggest a variety of adaptive responses relating involved a number of cellular processes elicited by frequent redox stress over several months. They may help to better understand inflammatory responses in redox related diseases and possibly allow uncovering new biomarkers of ROS-stress. Microarrays were used to analyze and investigate the biological effects of repeated exposure of cold physical plasma on human HaCaT keratinocytes. Using an argon plasma jet kinpen, regulated transcripts were analyzed and further described in Schmidt et al. (submittes): âLong-term exposure to cold plasma-generated ROS âan in vitro model for redox-related diseases of the skinâ. HaCaT keratinocytes exposed to plasma treated medium - time course
Project description:Investigation of gene expression in cultured human skin epithelial keratinocytes (HaCaT) following non-thermal plasma treatment for 60 s, compared to untreated control. Non-thermal atmospheric pressure plasma has recently gained attention in the field of biomedical and clinical applications. In the area of plasma medicine research one promising approach is to promote wound healing by stimulation of cells involved. To understand basic molecular and cellular mechanisms triggered by plasma treatment we investigated biological effects of an argon plasma jet (kinpen) on human epithelial skin cells. Consequently, whole-genome microarrays were used to analyze this interaction in detail and identified a statistically significant modification of 3,274 genes including 1,828 up- and 1,446 down-regulated genes. Particularly, plasma-treated cells are characterized by differential expression of a considerable number of genes involved in the response to stress. In this regard, we found a plasma-dependent regulation of oxidative stress answer and increased expression of enzymes of the antioxidative defense system (e.g. 91 oxidoreductases). Our results demonstrate that plasma induces cell reactions of stress-sensing but also of proliferative nature. Consistent with gene expression changes as well as Ingenuity Pathway Analysis prediction, we propose that stimulating doses of plasma may protect epithelial skin cells in wound healing by promoting proliferation and differentiation. In conclusion, gene expression profiling may become an important tool in identifying plasma-related changes of gene expression. Our results underline the enormous clinical potential of plasma as a biomedical tool for stimulation of epithelial skin cells We investigated biological effects of an argon plasma jet on HaCaTs. Microarray were used to analyzed this interaction in detail. The transcripts analyzed in this study are further described in Schmidt et al. (2013): Non-thermal plasma treatment is associated with changes in transcriptome of human epithelial skin cells. Accepted in Journal Free Radical Research A study using total RNA recovered from at least 8 non-thermal plasma treated samples (300 ms*M-BM-5l/cell) and untreated HaCaT controls.
Project description:Investigation of gene expression in cultured human skin epithelial keratinocytes (HaCaT) following non-thermal plasma treatment for 60 s, compared to untreated control. Non-thermal atmospheric pressure plasma has recently gained attention in the field of biomedical and clinical applications. In the area of plasma medicine research one promising approach is to promote wound healing by stimulation of cells involved. To understand basic molecular and cellular mechanisms triggered by plasma treatment we investigated biological effects of an argon plasma jet (kinpen) on human epithelial skin cells. Consequently, whole-genome microarrays were used to analyze this interaction in detail and identified a statistically significant modification of 3,274 genes including 1,828 up- and 1,446 down-regulated genes. Particularly, plasma-treated cells are characterized by differential expression of a considerable number of genes involved in the response to stress. In this regard, we found a plasma-dependent regulation of oxidative stress answer and increased expression of enzymes of the antioxidative defense system (e.g. 91 oxidoreductases). Our results demonstrate that plasma induces cell reactions of stress-sensing but also of proliferative nature. Consistent with gene expression changes as well as Ingenuity Pathway Analysis prediction, we propose that stimulating doses of plasma may protect epithelial skin cells in wound healing by promoting proliferation and differentiation. In conclusion, gene expression profiling may become an important tool in identifying plasma-related changes of gene expression. Our results underline the enormous clinical potential of plasma as a biomedical tool for stimulation of epithelial skin cells We investigated biological effects of an argon plasma jet on HaCaTs. Microarray were used to analyzed this interaction in detail. The transcripts analyzed in this study are further described in Schmidt et al. (2013): Non-thermal plasma treatment is associated with changes in transcriptome of human epithelial skin cells. Accepted in Journal Free Radical Research
Project description:Human Peptidoglycan Recognition Proteins (PGRPs) kill bacteria, likely by over-activating stress responses in bacteria. To gain insight into the mechanism of PGRP killing of Bacillus subtilis and bacterial defense against PGRP killing, gene expression in B. subtilis treated with a control protein (bovine serum albumin, BSA), human recombinant PGRP (PGLYRP4), gentamicin (aminoglycoside antibiotic), and CCCP (membrane potential decoupler) were compared. Each treatment induced unique and somewhat overlapping pattern of gene expression. PGRP highly increased expression of genes for oxidative and disulfide stress, detoxification and efflux of Cu, As, and Zn, several transporters, repair of damaged proteins and DNA, energy generation, histidine and cysteine synthesis, envelope lysis and remodeling, and other stress responses. PGRP also caused marked decrease in the expression of genes for phosphate uptake and utilization, Fe uptake, and motility. Gene expression microarray in B. subtilis exposed to a human bactericidal innate immunity protein, PGRP, showed induction of oxidative stress response and defense genes, with different expression pattern than B. subtilis exposed to an aminoglycoside antibiotic and a membrane potential decoupler. B. subtilis was treated with PGRP (human recombinant PGLYRP4), gentamicin, or CCCP (carbonyl cyanide 3-chlorophenylhydrazone). RNA was obtained from each culture and assayed for gene expression using custom whole genome Affymetrix 900513 GeneChip B. subtilis Genome Array. Each experiment was repeated 3 times.
Project description:Oxidative stress illustrates an imbalance between radical formation and removal. Frequent redox stress is critically involved in a variety of human pathologies including cancer, psoriasis, and chronic wounds. However, reactive species pursue a dual role being involved in signaling on the one hand and oxidative damage on the other. Using a HaCaT keratinocyte cell culture model, we here aimed at investigating the cellular and transcriptional response to periodic, low dose oxidative challenge over three months. Chronic redox stress was generated by frequent incubation with cold physical plasma treated cell culture medium. Using mRNA microarray technology, we found both acute ROS stress responses as well as numerous adaptions on the transcriptional level and over several weeks of redox challenge. This included an altered expression of 260 genes that function in inflammation and redox homeostasis, such as, signaling molecules, cytokines, and anti-oxidant enzymes. Apoptotic signaling was affected to a minor extend, especially in p53 down-stream targets. Strikingly, the anti-apoptotic heat shock protein HSP27 was strongly upregulated. These results suggest a variety of adaptive responses relating involved a number of cellular processes elicited by frequent redox stress over several months. They may help to better understand inflammatory responses in redox related diseases and possibly allow uncovering new biomarkers of ROS-stress. Microarrays were used to analyze and investigate the biological effects of repeated exposure of cold physical plasma on human HaCaT keratinocytes. Using an argon plasma jet kinpen, regulated transcripts were analyzed and further described in Schmidt et al. (submittes): “Long-term exposure to cold plasma-generated ROS –an in vitro model for redox-related diseases of the skin”.