Project description:The mammary glands of adult female mice were divided into ductal tissue and terminal end buds (TEBs). Basal and luminal epithelial cells were FACS sorted and nuclei extracted for 75bp paired-end ATAC-seq profiling using an Illumina NextSeq 500 sequencer.
Project description:The mammary glands of adult female mice were divided into ductal tissue and terminal end buds (TEBs). Basal and luminal epithelial cells were FACS sorted and RNA extracted for 75bp paired-end RNA-seq profiling using an Illumina NextSeq 500 sequencer.
Project description:Bone marrow-derived macrophages (BMDMs) were isolated from wild-type (WT) and Lipocalin 10 knockout (Lcn10-KO) mice and subjected to RNA-seq to investigate the transcriptional alterations associated with Lcn10 deficiency in macrophages. Total RNA was extracted from cultured BMDMs, and sequencing libraries were generated and sequenced on an Illumina NextSeq 500 platform using a single-end 85 base pair sequencing strategy. The resulting transcriptomic data were subsequently used for differential gene expression analysis and downstream pathway enrichment analyses to identify biological processes and signaling pathways potentially regulated by Lcn10 in macrophages.
Project description:Patients affected by type 1 diabetes are recruited in the departments of Diabetology and Healthy Volunteers (HV) are selected based on internal records in the same hospital. Total RNA from whole blood has been extracted following a two-step procedure. First, RNA from blood collected on PAX-Gene tubes has been extracted using Maxwell 16 LEV simplyRNA blood kit (Promega) following manufacturer recommendations and second b-globin, dominant RNA from red blood cells, has been removed using the GLOBINclear kit (Ambion) on extracted RNA. RNA sequencing has been performed from using the TruSeq Stranded mRNA preparation kit (Illumina) on 500 ng b-globin depleted RNA with a RNA Integrity Number > 8 (measured on Bioanalyzer following manufacturer recommendations), and then sequenced following a pair-end 2x75 bp protocol on NextSeq 500 or HiSeq 4000 (Illumina) at LIGAN Equipex (Lille, France).
Project description:Background: Mobile phones have become ubiquitous. There are concerns about possible health hazards posed by mobile phone-based radio frequency electromagnetic field (RF-EMF). Such exposure may be recognized as external stressor at molecular level which can be identified up by shot gun proteomics approach. Objective: To study molecular effects of mobile phone-based RF-EMF exposure using shot gun proteomics. Methods: 40 individuals (30 mobile phone users and 10 non-users) were recruited based on the inclusion and exclusion criteria. Mobile phone users had mobile phone usage of more than a year, an age group of 18-65 years with no major medical or surgical illness and normal results of hemogram, liver function tests (LFT), kidney function tests (KFT) and random/fasting sugars. Non-users had no mobile phone, Wi-Fi, and microwave usage. Both mobile phone users and non-users were non-alcoholic and non-smoker. RF-EMF exposure from mobile phone and miscellaneous sources was assessed using questionnaire. Calling-time duration per day was used as a measure of mobile phone exposure and the users were classified accordingly. Five biological replicates (1 set with both genders, 2 sets each of males and females) of proteomics experiments were designed. Human plasma samples were immunodepleted of high abundant proteins, concentrated, reduced, alkylated, digested, and labelled by i-TRAQ reagents for mass spectrometry analysis. Bioinformatics study was done on differentially expressed proteins for gene ontology (GO) classification, Protein-protein interactions (PPIs) and pathway enrichment. Results: The study group was quite young (median age 24 years). Pathway enrichment studies found Dopamine mediated receptor signaling, Adrenaline and Noradrenalin synthesis, Plasminogen activating cascade, Cholecystokinin receptor (CCKR) signaling, Gonadotropin-releasing hormone (GnRH) pathway and Vitamin-D signaling and its metabolism to be implicated. Conclusion: The involvement of the differentially expressed proteins in various signalling cascades have implications physiologically albeit large sample size studies incorporating non-users need to be conducted to draw out clear inferences.
Project description:Background: Mobile phones have become ubiquitous. There are concerns about possible health hazards posed by mobile phone-based radio frequency electromagnetic field (RF-EMF). Such exposure may be recognized as external stressor at molecular level which can be identified up by shot gun proteomics approach. Objective: To study molecular effects of mobile phone-based RF-EMF exposure using shot gun proteomics. Methods: 40 individuals (30 mobile phone users and 10 non-users) were recruited based on the inclusion and exclusion criteria. Mobile phone users had mobile phone usage of more than a year, an age group of 18-65 years with no major medical or surgical illness and normal results of hemogram, liver function tests (LFT), kidney function tests (KFT) and random/fasting sugars. Non-users had no mobile phone, Wi-Fi, and microwave usage. Both mobile phone users and non-users were non-alcoholic and non-smoker. RF-EMF exposure from mobile phone and miscellaneous sources was assessed using questionnaire. Calling-time duration per day was used as a measure of mobile phone exposure and the users were classified accordingly. Five biological replicates (1 set with both genders, 2 sets each of males and females) of proteomics experiments were designed. Human plasma samples were immunodepleted of high abundant proteins, concentrated, reduced, alkylated, digested, and labelled by i-TRAQ reagents for mass spectrometry analysis. Bioinformatics study was done on differentially expressed proteins for gene ontology (GO) classification, Protein-protein interactions (PPIs) and pathway enrichment. Results: The study group was quite young (median age 24 years). Pathway enrichment studies found Dopamine mediated receptor signaling, Adrenaline and Noradrenalin synthesis, Plasminogen activating cascade, Cholecystokinin receptor (CCKR) signaling, Gonadotropin-releasing hormone (GnRH) pathway and Vitamin-D signaling and its metabolism to be implicated. Conclusion: The involvement of the differentially expressed proteins in various signalling cascades have implications physiologically albeit large sample size studies incorporating non-users need to be conducted to draw out clear inferences.
Project description:Background: Mobile phones have become ubiquitous. There are concerns about possible health hazards posed by mobile phone-based radio frequency electromagnetic field (RF-EMF). Such exposure may be recognized as external stressor at molecular level which can be identified up by shot gun proteomics approach. Objective: To study molecular effects of mobile phone-based RF-EMF exposure using shot gun proteomics. Methods: 40 individuals (30 mobile phone users and 10 non-users) were recruited based on the inclusion and exclusion criteria. Mobile phone users had mobile phone usage of more than a year, an age group of 18-65 years with no major medical or surgical illness and normal results of hemogram, liver function tests (LFT), kidney function tests (KFT) and random/fasting sugars. Non-users had no mobile phone, Wi-Fi, and microwave usage. Both mobile phone users and non-users were non-alcoholic and non-smoker. RF-EMF exposure from mobile phone and miscellaneous sources was assessed using questionnaire. Calling-time duration per day was used as a measure of mobile phone exposure and the users were classified accordingly. Five biological replicates (1 set with both genders, 2 sets each of males and females) of proteomics experiments were designed. Human plasma samples were immunodepleted of high abundant proteins, concentrated, reduced, alkylated, digested, and labelled by i-TRAQ reagents for mass spectrometry analysis. Bioinformatics study was done on differentially expressed proteins for gene ontology (GO) classification, Protein-protein interactions (PPIs) and pathway enrichment. Results: The study group was quite young (median age 24 years). Pathway enrichment studies found Dopamine mediated receptor signaling, Adrenaline and Noradrenalin synthesis, Plasminogen activating cascade, Cholecystokinin receptor (CCKR) signaling, Gonadotropin-releasing hormone (GnRH) pathway and Vitamin-D signaling and its metabolism to be implicated. Conclusion: The involvement of the differentially expressed proteins in various signalling cascades have implications physiologically albeit large sample size studies incorporating non-users need to be conducted to draw out clear inferences.
Project description:Background: Mobile phones have become ubiquitous. There are concerns about possible health hazards posed by mobile phone-based radio frequency electromagnetic field (RF-EMF). Such exposure may be recognized as external stressor at molecular level which can be identified up by shot gun proteomics approach. Objective: To study molecular effects of mobile phone-based RF-EMF exposure using shot gun proteomics. Methods: 40 individuals (30 mobile phone users and 10 non-users) were recruited based on the inclusion and exclusion criteria. Mobile phone users had mobile phone usage of more than a year, an age group of 18-65 years with no major medical or surgical illness and normal results of hemogram, liver function tests (LFT), kidney function tests (KFT) and random/fasting sugars. Non-users had no mobile phone, Wi-Fi, and microwave usage. Both mobile phone users and non-users were non-alcoholic and non-smoker. RF-EMF exposure from mobile phone and miscellaneous sources was assessed using questionnaire. Calling-time duration per day was used as a measure of mobile phone exposure and the users were classified accordingly. Five biological replicates (1 set with both genders, 2 sets each of males and females) of proteomics experiments were designed. Human plasma samples were immunodepleted of high abundant proteins, concentrated, reduced, alkylated, digested, and labelled by i-TRAQ reagents for mass spectrometry analysis. Bioinformatics study was done on differentially expressed proteins for gene ontology (GO) classification, Protein-protein interactions (PPIs) and pathway enrichment. Results: The study group was quite young (median age 24 years). Pathway enrichment studies found Dopamine mediated receptor signaling, Adrenaline and Noradrenalin synthesis, Plasminogen activating cascade, Cholecystokinin receptor (CCKR) signaling, Gonadotropin-releasing hormone (GnRH) pathway and Vitamin-D signaling and its metabolism to be implicated. Conclusion: The involvement of the differentially expressed proteins in various signalling cascades have implications physiologically albeit large sample size studies incorporating non-users need to be conducted to draw out clear inferences.
Project description:Background: Mobile phones have become ubiquitous. There are concerns about possible health hazards posed by mobile phone-based radio frequency electromagnetic field (RF-EMF). Such exposure may be recognized as external stressor at molecular level which can be identified up by shot gun proteomics approach. Objective: To study molecular effects of mobile phone-based RF-EMF exposure using shot gun proteomics. Methods: 40 individuals (30 mobile phone users and 10 non-users) were recruited based on the inclusion and exclusion criteria. Mobile phone users had mobile phone usage of more than a year, an age group of 18-65 years with no major medical or surgical illness and normal results of hemogram, liver function tests (LFT), kidney function tests (KFT) and random/fasting sugars. Non-users had no mobile phone, Wi-Fi, and microwave usage. Both mobile phone users and non-users were non-alcoholic and non-smoker. RF-EMF exposure from mobile phone and miscellaneous sources was assessed using questionnaire. Calling-time duration per day was used as a measure of mobile phone exposure and the users were classified accordingly. Five biological replicates (1 set with both genders, 2 sets each of males and females) of proteomics experiments were designed. Human plasma samples were immunodepleted of high abundant proteins, concentrated, reduced, alkylated, digested, and labelled by i-TRAQ reagents for mass spectrometry analysis. Bioinformatics study was done on differentially expressed proteins for gene ontology (GO) classification, Protein-protein interactions (PPIs) and pathway enrichment. Results: The study group was quite young (median age 24 years). Pathway enrichment studies found Dopamine mediated receptor signaling, Adrenaline and Noradrenalin synthesis, Plasminogen activating cascade, Cholecystokinin receptor (CCKR) signaling, Gonadotropin-releasing hormone (GnRH) pathway and Vitamin-D signaling and its metabolism to be implicated. Conclusion: The involvement of the differentially expressed proteins in various signalling cascades have implications physiologically albeit large sample size studies incorporating non-users need to be conducted to draw out clear inferences.