1) The document discusses wound healing and fibrosis, providing background on the normal wound healing process and how uncontrolled wound healing can lead to fibrosis. It describes key cell types, proteins, and signaling pathways involved in wound healing and fibrosis.
2) Three case studies are summarized that used RT2 Profiler PCR Arrays to study fibrosis. One looked at IL-17 signaling and collagen expression in scleroderma fibroblasts. Another examined the role of SOCS3 in left ventricular remodeling after myocardial infarction. A third studied membrane type 1-matrix metalloproteinase induction and its relationship to left ventricular remodeling and fibrosis.
3) RT2 Profiler PCR Arrays are introduced as pathway-focused gene expression profiling tools for
Objective: To probe into the influence of miR-21 on the proliferation as well as apoptosis of oral squamous cell carcinoma (OSCC) and its causative role.
Study Design: We adopted microarray for detecting the differentially expressed genes in OSCC tumor tis-sues and paracancerous tissues. We assessed the link of miR-21 expression with tumor size, lymph node metastasis, and tumor differentiation. We employed CCK-8 and EdU assay for detecting the impact of miR-21 inhibitor and miR-21 mimic on Cal-27 cell proliferation, as well as TUNEL and AnnexinV-FITC/PI double staining for detecting miR-21 expression on cell apoptosis. We forecasted the possible target of miR-21 via TargetScan, as well as detected the interaction of miR-21 with PTEN via luciferase reporter experiment. The function of miR-21 expression in PTEN signaling pathway was monitored via western blot. We constructed PTEN overexpression plasmid and conducted rescue experiment to evaluate overexpressed PTEN on miR-21–induced proliferation.
Results: Microarray and RT-qPCR indicated that miR-21 expression increased demonstrably in OSCC. Subsequently, statistical analysis showed that miR-21 expression was plainly correlated with tumor size, lymph node metastasis, tumor differentiation, and smoking history. CCK-8 and EdU method exhibited that miR-21 mimics manifestly promoted Cal-27 cell proliferation, while miR-21 inhibitor blatantly inhibited Cal-27 cell proliferation. TUNEL and V-FITC/PI double staining assay showed that miR-21 inhibitor conspicuously promoted Cal-27 cell apoptosis. CCK-8 and EdU assay exhibited that overexpressed PTEN abolished the pro-proliferation influence of miR-21 mimic. TUNEL and V-FITC/PI experiments pointed out that knocking down PTEN abrogated the pro-apoptosis impact of miR-21 inhibitor.
Conclusion: miR-21 contributes to OSCC cell proliferation via targeting PTEN and inhibits its apoptosis.
Keywords: Akt/PKB signaling pathway; apoptosis; biomarkers, tumor; carcinoma, squamous cell; cell line, tumor; cell proliferation; microRNAs; miR-21; miRNA-21; mouth neoplasms; oral cancer; oral squamous cell carcinoma; proliferation; real time PCR
Objective: Ischemia-reperfusion (I/R) leads to reactive oxygen species formation and cell death in kidney tissue with injury and organ transplantation. Simvastatin (SIM) is an antioxidant, anti-inflammatory, and anticoagulant agent. Alterations in I/R-induced acute kidney injury model with SIM treatment were analyzed.
Study Design: Wistar rats (n=28) were grouped into Sham, Ischemia, I/R, and I/R+SIM treated. Left rat kidney renal vessels were clamped for 60 minutes for ischemia, and the I/R group had 6 hours of reperfusion. 10 mg/kg SIM was given orally for 28 days. MDA, GSH, and MPO were analyzed. Kidney tissues were paraffin embedded, and primary antibodies TNF-α and caspase-3 were applied for immunohistochemistry.
Results: In the I/R group, intense inflammatory cell infiltration around the vessels and necrosis in the glomerular structures were observed. In the treated group, proximal and distal tubular cells were found to be close to normal. Immunoexpression of caspase-3 in the ischemia group was positive in degenerative glomeruli. In the treated group, TNF-α expression was negative in the glomerular structures. MDA and MPO levels were significantly increased in ischemia and I/R.
Conclusion: We suggest that SIM treatment improved kidney tissue structure and function in a model of I/R injury.
Keywords: caspase-3; immunohistochemistry; ischemia/reperfusion; kidney; MPO; simvastatin
Objective: To probe into the influence of miR-21 on the proliferation as well as apoptosis of oral squamous cell carcinoma (OSCC) and its causative role.
Study Design: We adopted microarray for detecting the differentially expressed genes in OSCC tumor tis-sues and paracancerous tissues. We assessed the link of miR-21 expression with tumor size, lymph node metastasis, and tumor differentiation. We employed CCK-8 and EdU assay for detecting the impact of miR-21 inhibitor and miR-21 mimic on Cal-27 cell proliferation, as well as TUNEL and AnnexinV-FITC/PI double staining for detecting miR-21 expression on cell apoptosis. We forecasted the possible target of miR-21 via TargetScan, as well as detected the interaction of miR-21 with PTEN via luciferase reporter experiment. The function of miR-21 expression in PTEN signaling pathway was monitored via western blot. We constructed PTEN overexpression plasmid and conducted rescue experiment to evaluate overexpressed PTEN on miR-21–induced proliferation.
Results: Microarray and RT-qPCR indicated that miR-21 expression increased demonstrably in OSCC. Subsequently, statistical analysis showed that miR-21 expression was plainly correlated with tumor size, lymph node metastasis, tumor differentiation, and smoking history. CCK-8 and EdU method exhibited that miR-21 mimics manifestly promoted Cal-27 cell proliferation, while miR-21 inhibitor blatantly inhibited Cal-27 cell proliferation. TUNEL and V-FITC/PI double staining assay showed that miR-21 inhibitor conspicuously promoted Cal-27 cell apoptosis. CCK-8 and EdU assay exhibited that overexpressed PTEN abolished the pro-proliferation influence of miR-21 mimic. TUNEL and V-FITC/PI experiments pointed out that knocking down PTEN abrogated the pro-apoptosis impact of miR-21 inhibitor.
Conclusion: miR-21 contributes to OSCC cell proliferation via targeting PTEN and inhibits its apoptosis.
Keywords: Akt/PKB signaling pathway; apoptosis; biomarkers, tumor; carcinoma, squamous cell; cell line, tumor; cell proliferation; microRNAs; miR-21; miRNA-21; mouth neoplasms; oral cancer; oral squamous cell carcinoma; proliferation; real time PCR
Objective: Ischemia-reperfusion (I/R) leads to reactive oxygen species formation and cell death in kidney tissue with injury and organ transplantation. Simvastatin (SIM) is an antioxidant, anti-inflammatory, and anticoagulant agent. Alterations in I/R-induced acute kidney injury model with SIM treatment were analyzed.
Study Design: Wistar rats (n=28) were grouped into Sham, Ischemia, I/R, and I/R+SIM treated. Left rat kidney renal vessels were clamped for 60 minutes for ischemia, and the I/R group had 6 hours of reperfusion. 10 mg/kg SIM was given orally for 28 days. MDA, GSH, and MPO were analyzed. Kidney tissues were paraffin embedded, and primary antibodies TNF-α and caspase-3 were applied for immunohistochemistry.
Results: In the I/R group, intense inflammatory cell infiltration around the vessels and necrosis in the glomerular structures were observed. In the treated group, proximal and distal tubular cells were found to be close to normal. Immunoexpression of caspase-3 in the ischemia group was positive in degenerative glomeruli. In the treated group, TNF-α expression was negative in the glomerular structures. MDA and MPO levels were significantly increased in ischemia and I/R.
Conclusion: We suggest that SIM treatment improved kidney tissue structure and function in a model of I/R injury.
Keywords: caspase-3; immunohistochemistry; ischemia/reperfusion; kidney; MPO; simvastatin
BACKGROUND: Sequential Epstein-Barr virus (EBV)–positive B cell lymphoma to the initial diagnosis of angioimmunoblastic T cell lymphoma (AITL) is very rare, the exact mechanism and standard therapy of which is still being explored. CASE: A 50-year-old man was admitted to our hospital in January 2014 with a three-week history of enlargement of multiple lymph nodes. His initial pathological evaluation indicated AILT. The reactivation of EBV was observed during the immunosuppression therapy for AITL, accompanied by onset of subcutaneous nodules proven to be EBV-positive diffuse large B cell lymphoma (DLBCL) based on the pathological findings of rebiopsy. The patient was successfully treated with chidamide, a histone deacetylase (HDAC) inhibitor, and rituximab.
Conclusion: The sufficient surveillance for serum EBV and repeat biopsy is necessary for patients with AITL, and this treatment modality may become an active option.
Keywords: angioimmunoblastic T cell lymphoma, Epstein-Barr virus, HDAC inhibitor, non-Hodgkin lymphoma, peripheral T cell lymphoma
Aim of this ppt presentation:
To understand the standard of care for both GBM and anaplastic glioma.
To know what is the new advances and modifications to the standard of care?
Contents:
Introduction: 2 slides.
GBM:
Epidemiology: 1 slide.
Molecular biology & New trends: 5 slides
EORTC/NCIC trial: 10 slides.
MGMT: 1 slide.
Evidence-based medicine: 6 slides.
Avastin in GBM: 2 slides.
Novocure (TTF): 2 slides.
Gliadel (BCNU) wafers: 1 slide.
Anaplastic astrocytoma: 7 slides
Take home message.
Conferència a càrrec d'Enriqueta Felip. Oncòloga de l'Hospital Vall d'Hebron de Barcelona. Líder en la investigació de càncer de pulmó al VHIO. Membre de la Junta Directiva de la Societat Espanyola d'Oncologia Mèdica i de l'European Society for Medical Oncology. En el marc de la Jornada "Els nous reptes de la Medicina de Precisió" organitzada el 12 de novembre per la Societat Catalana de Gestió Sanitària
Stem Cells in A New Era of Cell based Therapies - Creative BiolabsCreative-Biolabs
A stem cell can replicate itself or differentiate into cells that carry out the specific functions of the body. The application of stem cells in regenerative medicine and disease therapeutics is one of the most exciting advances in medical science today. In cell-based therapies, stem cells may play two roles. The first role is as drug-delivery vehicles. The second role is as therapeutic agents themselves. Stem cells also offer opportunities for scientific advances that go far beyond cell-based therapies. Creative Biolabs is dedicated to facilitate the research of stem cells in both basic science and therapeutics development. Please contact us if you are interested in our services or products.
Developing a Rapid Clinical Sequencing System to Classify Meningioma: Meet th...QIAGEN
Meningioma’s display a broad spectrum of clinical, histological and cytogenetic features even within the same WHO grade often posing a challenge for classification and prognostic stratification. In this webinar, we will describe our experience of using targeted amplicon sequencing to develop rapid clinical sequencing system to identify and confirm the meningioma genotype in just two weeks. In addition the details of the three meningioma categories and the genes involved will be discussed.
Direct Sanger CE Sequencing of Individual Ampliseq Cancer Panel Targets from ...Thermo Fisher Scientific
The introduction of defined Ion AmpliSeq™ panels for detection and characterization of actionable mutations occurring in tumor tissue has the potential to revolutionize translational oncology research. The Ion Ampliseq™ cancer hot spot panel version 2 (CHP v2) by Ion Torrent includes 207 actionable sequences from a single target and mutation targets present in 50 genes and the more comprehensive Ion Oncomine™ cancer panel (OCP) developed by Life Technologies Compendia Bioscience™ contains over 2000 mutations. A hallmark of these Ion Torrent Ampliseq cancer panels is the low amount of input DNA needed which is critical when the clinical specimen material is limited such as with fine needle biopsy or FFPE samples. Typically, 10 ng of DNA obtained from these sources is sufficient to produce informative sequencing data. Often, cancer-causing or promoting mutations are detected at relatively low allele frequencies like 10-20 % compared to the major normal allele. Many researchers wish to verify these findings of low frequency mutations by an orthologous method such as traditional dye-fluorescent Sanger sequencing on a capillary electrophoresis (CE) instrument such as the Applied Biosystems 3500 genetic analyzer. To that end, we have developed a workflow that enables the amplification and traditional Sanger sequencing of individual Ion AmpliSeq targets directly from the AmpliSeq library starting material.
The method requires a retainer of 1 μl (~ 5%) of the original AmpliSeq preamplification material. A dilution of this aliquot is used as template source for individualized PCR/sequencing reactions. We show that a random selection of 48 targets from the CHPv2 panel could be successfully amplified and Sanger-sequenced from an Ion Torrent Ampliseq library originally prepared from 10 ng of FFPE
DNA. Furthermore, we show the successful Sanger-re-sequencing of all individual 24 targets covering the TP53 exons from the same sample processed and pre-amplified with the OncoMine AmpliSeq panel.
Taken together, this method will enable researchers to reflex-test potential mutations of interest from very material-limited specimen using Sanger CE sequencing
Treating cancer effectively requires an understanding of the molecular alterations driving each patient’s tumor. Targeted sequencing efforts that characterize prevalent somatic alterations and require limited sample input may provide an effective diagnostic approach. Herein, we describe the design and characterization of the Oncomine™ Cancer Research Panel (OCP) that includes recurrent somatic alterations in solid tumors derived from the Oncomine™ cancer database. Using Ion AmpliSeq™ technology, we designed a DNA panel that includes assays for 73 oncogenes with 1,826 recurrent hotspot mutations, 26 tumor suppressor genes enriched for deleterious mutations, as well as 75 genes subject to recurrent focal copy gain or loss. A complementary RNA panel includes 183 assays for relevant gene fusions involving 22 fusion driver genes. Recommended sample inputs were 10 ng of nucleic acid per pool. Sequencing libraries were analyzed on an Ion Torrent™ Personal Genome Machine™. Initial testing revealed an average read depth of > 1,500X with > 95% uniformity and on target frequency. The panel was shown to reliably detect known hotspots, insertions/deletions, gene copy changes, and gene fusions in molecular standards, cell lines and formalin-fixed paraffin embedded samples. Retrospective analysis of large sample cohorts has been completed and the results of analysis of 100 lung cancer and 100 prostate cancer cases will be summarized. In addition, a prospective cohort of 100 samples from the University of Michigan Molecular Diagnostics laboratory was profiled with OCP. Overall, we achieved >95% sensitivity and specificity for detection of KRAS, EGFR and BRAF mutations and ALK gene fusions.
Objective: Tongue squamous cell carcinoma (TSCC) is a prominent type of oral cancer. Despite the numerous research studies on SCC and microRNAs (miRs), the relation between TSCC and miR-135b-5p is poorly discussed. This experiment aims to find out the possible effect of miR-135b-5p on TSCC with the network of its downstream genes.
Study Design: TSCC tissues and adjacent normal tissues were harvested. Then, expression of miR-135b-5p and AT-rich interactive domain‑containing protein 1A gene (ARID1A) and the phosphatidyl inositol 3-kinase/protein kinase B (PI3K/AKT) pathway was analyzed. After the transfection of miR-135b-5p inhibitor and its negative control into TSCC cells, functional assays were employed to measure cell proliferation, apoptosis, and cycle. Next, the target relation between miR-135b-5p and ARID1A was confirmed. In addition, the fact that miR-135b-5p promoted TSCC development via mediating ARID1A was demonstrated by functional rescue experiment.
Results: miR-135b-5p was upregulated in TSCC tissues and cells, while ARID1A was suppressed (p< 0.05). Silenced miR-135b-5p discouraged TSCC cell proliferation, improved apoptosis, induced cell cycle arrest, and increased ARID1A expression while inactivating the PI3K/AKT axis (p<0.05). Furthermore, knockdown of ARID1A reversed the impacts on TSCC cell proliferation and apoptosis exerted by silencing miR-135b-5p.
Conclusion: This research supported that silenced miR-135b-5p impeded TSCC proliferation and apoptosis by promoting ARID1A and inactivating the PI3K/AKT axis, which may provide some indications for TSCC alleviation.
Keywords: apoptosis; ARID1A; ARID1A protein, human; carcinoma, squamous cell; cell line, tumor; cell proliferation; drug resistance, neoplasm; microRNA-135b-5p; microRNAs; PI3K/AKT pathway; neoplasm metastasis; neoplastic stem cells; proliferation; protein binding; tongue; tongue squamous cell carcinoma
In this presentation we showcase the latest advancements in myeloid genomic profiling: The Ion Torrent Oncomine Myeloid Assay GX.
Learn how this solution addresses key challenges in myeloid molecular testing and see recent data from the University of Pennsylvania.
Learn more at www.oncomine.com/myeloid
In this OncomineWorld 2021 presentation, learn how next-generation sequencing is playing a crucial role in cancer research. To watch the full presentation, visit www.oncomine.com/events.
Next-Generation Sequencing Clinical Research Milestones InfographicQIAGEN
DNA mutations have been implicated in several diseases, particularly cancer. NGS presents an ideal technology to efficiently profile the multitude of mutations in a high throughput manner. In 2001 the first draft of human genome was published. Since then many major milestones have been reached. Do you know when PIK3CA was identified in colon cancer? When was Olaparib for ovarian cancer treatment? This infographic traces the major clinical research milestones starting from the first draft of the human genome.
Sequencing 60,000 Samples: An Innovative Large Cohort Study for Breast Cancer...QIAGEN
This slidedeck focuses on the design of a large cohort study for assessing breast cancer risk and how an innovative digital sequencing approach is able to solve the previously unmet challenges of this type of NGS study design. Our speaker, Dr. Fergus J. Couch of the Mayo Clinic, presents on the design of this NCI-funded project, which comprises the sequencing of 60,000 samples to assess the risk of breast cancer through association with targeted genes. The design and size of the study requires an accurate, robust and high-throughput sequencing method. The investigators are using a digital DNA sequencing approach from QIAGEN that incorporates molecular barcodes to tag and remove PCR duplicates and increase NGS assay sensitivity. The approach also uses proprietary chemistry that enables uniform sequencing to efficiently utilize sequencing power and deliver optimized results.
BACKGROUND: Sequential Epstein-Barr virus (EBV)–positive B cell lymphoma to the initial diagnosis of angioimmunoblastic T cell lymphoma (AITL) is very rare, the exact mechanism and standard therapy of which is still being explored. CASE: A 50-year-old man was admitted to our hospital in January 2014 with a three-week history of enlargement of multiple lymph nodes. His initial pathological evaluation indicated AILT. The reactivation of EBV was observed during the immunosuppression therapy for AITL, accompanied by onset of subcutaneous nodules proven to be EBV-positive diffuse large B cell lymphoma (DLBCL) based on the pathological findings of rebiopsy. The patient was successfully treated with chidamide, a histone deacetylase (HDAC) inhibitor, and rituximab.
Conclusion: The sufficient surveillance for serum EBV and repeat biopsy is necessary for patients with AITL, and this treatment modality may become an active option.
Keywords: angioimmunoblastic T cell lymphoma, Epstein-Barr virus, HDAC inhibitor, non-Hodgkin lymphoma, peripheral T cell lymphoma
Aim of this ppt presentation:
To understand the standard of care for both GBM and anaplastic glioma.
To know what is the new advances and modifications to the standard of care?
Contents:
Introduction: 2 slides.
GBM:
Epidemiology: 1 slide.
Molecular biology & New trends: 5 slides
EORTC/NCIC trial: 10 slides.
MGMT: 1 slide.
Evidence-based medicine: 6 slides.
Avastin in GBM: 2 slides.
Novocure (TTF): 2 slides.
Gliadel (BCNU) wafers: 1 slide.
Anaplastic astrocytoma: 7 slides
Take home message.
Conferència a càrrec d'Enriqueta Felip. Oncòloga de l'Hospital Vall d'Hebron de Barcelona. Líder en la investigació de càncer de pulmó al VHIO. Membre de la Junta Directiva de la Societat Espanyola d'Oncologia Mèdica i de l'European Society for Medical Oncology. En el marc de la Jornada "Els nous reptes de la Medicina de Precisió" organitzada el 12 de novembre per la Societat Catalana de Gestió Sanitària
Stem Cells in A New Era of Cell based Therapies - Creative BiolabsCreative-Biolabs
A stem cell can replicate itself or differentiate into cells that carry out the specific functions of the body. The application of stem cells in regenerative medicine and disease therapeutics is one of the most exciting advances in medical science today. In cell-based therapies, stem cells may play two roles. The first role is as drug-delivery vehicles. The second role is as therapeutic agents themselves. Stem cells also offer opportunities for scientific advances that go far beyond cell-based therapies. Creative Biolabs is dedicated to facilitate the research of stem cells in both basic science and therapeutics development. Please contact us if you are interested in our services or products.
Developing a Rapid Clinical Sequencing System to Classify Meningioma: Meet th...QIAGEN
Meningioma’s display a broad spectrum of clinical, histological and cytogenetic features even within the same WHO grade often posing a challenge for classification and prognostic stratification. In this webinar, we will describe our experience of using targeted amplicon sequencing to develop rapid clinical sequencing system to identify and confirm the meningioma genotype in just two weeks. In addition the details of the three meningioma categories and the genes involved will be discussed.
Direct Sanger CE Sequencing of Individual Ampliseq Cancer Panel Targets from ...Thermo Fisher Scientific
The introduction of defined Ion AmpliSeq™ panels for detection and characterization of actionable mutations occurring in tumor tissue has the potential to revolutionize translational oncology research. The Ion Ampliseq™ cancer hot spot panel version 2 (CHP v2) by Ion Torrent includes 207 actionable sequences from a single target and mutation targets present in 50 genes and the more comprehensive Ion Oncomine™ cancer panel (OCP) developed by Life Technologies Compendia Bioscience™ contains over 2000 mutations. A hallmark of these Ion Torrent Ampliseq cancer panels is the low amount of input DNA needed which is critical when the clinical specimen material is limited such as with fine needle biopsy or FFPE samples. Typically, 10 ng of DNA obtained from these sources is sufficient to produce informative sequencing data. Often, cancer-causing or promoting mutations are detected at relatively low allele frequencies like 10-20 % compared to the major normal allele. Many researchers wish to verify these findings of low frequency mutations by an orthologous method such as traditional dye-fluorescent Sanger sequencing on a capillary electrophoresis (CE) instrument such as the Applied Biosystems 3500 genetic analyzer. To that end, we have developed a workflow that enables the amplification and traditional Sanger sequencing of individual Ion AmpliSeq targets directly from the AmpliSeq library starting material.
The method requires a retainer of 1 μl (~ 5%) of the original AmpliSeq preamplification material. A dilution of this aliquot is used as template source for individualized PCR/sequencing reactions. We show that a random selection of 48 targets from the CHPv2 panel could be successfully amplified and Sanger-sequenced from an Ion Torrent Ampliseq library originally prepared from 10 ng of FFPE
DNA. Furthermore, we show the successful Sanger-re-sequencing of all individual 24 targets covering the TP53 exons from the same sample processed and pre-amplified with the OncoMine AmpliSeq panel.
Taken together, this method will enable researchers to reflex-test potential mutations of interest from very material-limited specimen using Sanger CE sequencing
Treating cancer effectively requires an understanding of the molecular alterations driving each patient’s tumor. Targeted sequencing efforts that characterize prevalent somatic alterations and require limited sample input may provide an effective diagnostic approach. Herein, we describe the design and characterization of the Oncomine™ Cancer Research Panel (OCP) that includes recurrent somatic alterations in solid tumors derived from the Oncomine™ cancer database. Using Ion AmpliSeq™ technology, we designed a DNA panel that includes assays for 73 oncogenes with 1,826 recurrent hotspot mutations, 26 tumor suppressor genes enriched for deleterious mutations, as well as 75 genes subject to recurrent focal copy gain or loss. A complementary RNA panel includes 183 assays for relevant gene fusions involving 22 fusion driver genes. Recommended sample inputs were 10 ng of nucleic acid per pool. Sequencing libraries were analyzed on an Ion Torrent™ Personal Genome Machine™. Initial testing revealed an average read depth of > 1,500X with > 95% uniformity and on target frequency. The panel was shown to reliably detect known hotspots, insertions/deletions, gene copy changes, and gene fusions in molecular standards, cell lines and formalin-fixed paraffin embedded samples. Retrospective analysis of large sample cohorts has been completed and the results of analysis of 100 lung cancer and 100 prostate cancer cases will be summarized. In addition, a prospective cohort of 100 samples from the University of Michigan Molecular Diagnostics laboratory was profiled with OCP. Overall, we achieved >95% sensitivity and specificity for detection of KRAS, EGFR and BRAF mutations and ALK gene fusions.
Objective: Tongue squamous cell carcinoma (TSCC) is a prominent type of oral cancer. Despite the numerous research studies on SCC and microRNAs (miRs), the relation between TSCC and miR-135b-5p is poorly discussed. This experiment aims to find out the possible effect of miR-135b-5p on TSCC with the network of its downstream genes.
Study Design: TSCC tissues and adjacent normal tissues were harvested. Then, expression of miR-135b-5p and AT-rich interactive domain‑containing protein 1A gene (ARID1A) and the phosphatidyl inositol 3-kinase/protein kinase B (PI3K/AKT) pathway was analyzed. After the transfection of miR-135b-5p inhibitor and its negative control into TSCC cells, functional assays were employed to measure cell proliferation, apoptosis, and cycle. Next, the target relation between miR-135b-5p and ARID1A was confirmed. In addition, the fact that miR-135b-5p promoted TSCC development via mediating ARID1A was demonstrated by functional rescue experiment.
Results: miR-135b-5p was upregulated in TSCC tissues and cells, while ARID1A was suppressed (p< 0.05). Silenced miR-135b-5p discouraged TSCC cell proliferation, improved apoptosis, induced cell cycle arrest, and increased ARID1A expression while inactivating the PI3K/AKT axis (p<0.05). Furthermore, knockdown of ARID1A reversed the impacts on TSCC cell proliferation and apoptosis exerted by silencing miR-135b-5p.
Conclusion: This research supported that silenced miR-135b-5p impeded TSCC proliferation and apoptosis by promoting ARID1A and inactivating the PI3K/AKT axis, which may provide some indications for TSCC alleviation.
Keywords: apoptosis; ARID1A; ARID1A protein, human; carcinoma, squamous cell; cell line, tumor; cell proliferation; drug resistance, neoplasm; microRNA-135b-5p; microRNAs; PI3K/AKT pathway; neoplasm metastasis; neoplastic stem cells; proliferation; protein binding; tongue; tongue squamous cell carcinoma
In this presentation we showcase the latest advancements in myeloid genomic profiling: The Ion Torrent Oncomine Myeloid Assay GX.
Learn how this solution addresses key challenges in myeloid molecular testing and see recent data from the University of Pennsylvania.
Learn more at www.oncomine.com/myeloid
In this OncomineWorld 2021 presentation, learn how next-generation sequencing is playing a crucial role in cancer research. To watch the full presentation, visit www.oncomine.com/events.
Next-Generation Sequencing Clinical Research Milestones InfographicQIAGEN
DNA mutations have been implicated in several diseases, particularly cancer. NGS presents an ideal technology to efficiently profile the multitude of mutations in a high throughput manner. In 2001 the first draft of human genome was published. Since then many major milestones have been reached. Do you know when PIK3CA was identified in colon cancer? When was Olaparib for ovarian cancer treatment? This infographic traces the major clinical research milestones starting from the first draft of the human genome.
Sequencing 60,000 Samples: An Innovative Large Cohort Study for Breast Cancer...QIAGEN
This slidedeck focuses on the design of a large cohort study for assessing breast cancer risk and how an innovative digital sequencing approach is able to solve the previously unmet challenges of this type of NGS study design. Our speaker, Dr. Fergus J. Couch of the Mayo Clinic, presents on the design of this NCI-funded project, which comprises the sequencing of 60,000 samples to assess the risk of breast cancer through association with targeted genes. The design and size of the study requires an accurate, robust and high-throughput sequencing method. The investigators are using a digital DNA sequencing approach from QIAGEN that incorporates molecular barcodes to tag and remove PCR duplicates and increase NGS assay sensitivity. The approach also uses proprietary chemistry that enables uniform sequencing to efficiently utilize sequencing power and deliver optimized results.
Assessment of immunomolecular_expression_and_prognostic_role_of_tlr7_among_pa...dr.Ihsan alsaimary
Dr. ihsan edan abdulkareem alsaimary
PROFESSOR IN MEDICAL MICROBIOLOGY AND MOLECULAR IMMUNOLOGY
ihsanalsaimary@gmail.com
mobile : 009647801410838
university of basrah - college of medicine - basrah -IRAQ
Src jbbr-20-120 Dr. ihsan edan abdulkareem alsaimary PROFESSOR IN MEDICAL M...dr.Ihsan alsaimary
Dr. ihsan edan abdulkareem alsaimary
PROFESSOR IN MEDICAL MICROBIOLOGY AND MOLECULAR IMMUNOLOGY
ihsanalsaimary@gmail.com
mobile : 009647801410838
university of basrah - college of medicine - basrah -IRAQ
Integrating Disruptive Technologies Into Translational Research Hinxton Hal...Mike Romanos
Introduction to a session with the same title. Addresses the challenges in drug discovery and how disruptive technologies can help. Focusses on use of RNAi and stem cells in translational studies
Use of TEG® in Acute Traumatic Brain Injurykathleenmccann
Acute traumatic coagulopathy complicate 25 – 75% of cases of traumatic brain injury, incidence increasing with severity of injury. The complex processes that cause this are a culmination of the activation of anticoagulant and fibrinolytic pathways. Much has been published about the clinical outcomes of brain injury and the associated coagulopathy with platelet dysfunction. The mechanism, however, is still not understood.
Clinical management of these patients to date relies on traditional serum based screens: PT, INR, PTT, platelet counts, fibrinogen, and D-dimer assays. These values do not reflect whole blood hemostasis nor do they reflect a cell based model of hemostasis. Thromboelastography with platelet mapping, however, provides real-time data on:
Coagulation pathway function
Fibrinogen function
Platelet function
Fibrinolysis
We purport the expanded use of TEG® as a viscoelastic point-of-care test of whole blood which more accurately reflects hemostasis. It determines the hemostatic integrity of whole blood in vitro by giving real-time information on thrombus initiation, amplification, propagation, and termination. It has long been used in management in transplant patients and during cardiac surgery. It has recently become increasingly used in the management of trauma patients, including those presenting with traumatic brain injury.
We show that TEG® is useful not only in guiding treatment, but also in studying the mechanisms underlying acute traumatic coagulopathy and platelet dysfunction in cases of hemorrhagic brain injury, in both the animal model and in the clinical setting.
Webinar analyzing complex genomic variants in somatic cancer Lisa Owen
Next generation sequencing (NGS) technologies facilitate the accurate detection of genetic and genomic variants. Yet the process of analyzing and classifying more complex alterations remains challenging.
In this slide deck, which is a companion to the webinar presented on February 21, 2019 and located here (https://www.pieriandx.com/analyzingcomplexgenomicvariants) you’ll learn how to analyze complex genomic alterations, such as gene fusions, splice-site mutations, and co-occurring variants within the context of somatic cancer.
1. Sample & Assay Technologies
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Uncovering the mechanisms of wound
healing and fibrosis
2. Sample & Assay Technologies
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Title, Location, Date
2
3. Sample & Assay Technologies
Background: process of wound healing
Wound healing — a 4-step process
Step 1: Hemostasis (clotting)
Step 2: Inflammation
Vasoconstriction
Enhanced blood vessel permeability due to release
of histamine and other factors
Coagulation cascade is initiated by interaction of
coagulation factor FVII with TF
Von Willebrand factor helps platelets bind at wound
sites, where they activate and degranulate
Step 3: Proliferative phase
Inflammatory cells infiltrate (PMN, macrophages)
and kill any microbes accompanying the injury
Leukocytes produce cytokines, chemokines, and
growth factors, some of which (IL-1beta, TGF-beta,
TNF) recruit fibroblasts
Step 4: Wound closing/tissue remodeling
Fibroblasts are recruited and activated, and secrete Wound contraction via myofibroblasts at the edges
ECM components (type III collagen, fibronectin)
Formation of granulation tissue (fibroblasts,
inflammatory cells, new blood vessels, fibronectin,
hyaluronan, collagen, endothelial cells)
Type III collagen is replaced by Type I, and fibers
are rearranged and crosslinked.
Remodeling will continue for weeks to months.
Epithelialization
Title, Location, Date
3
4. Sample & Assay Technologies
Backgrounds: wound healing and fibrosis
Uncontrolled wound healing response results in fibrosis
Fibrosis develops if any stage in the tissue repair program is dysregulated.
Chronic inflammation
The tissue-damaging agent is not removed
The repair process is not regulated properly
Review, Integrating mechanisms of pulmonary fibrosis. JEM Vol. 208, No. 7
Title, Location, Date
4
6. Sample & Assay Technologies
Background: Fibrosis and wound healing
Fibrosis affects a wide range of tissues, including:
Lungs (idiopathic pulmonary fibrosis, cystic fibrosis)
Heart (post-myocardial infarction, endomyocardial
fibrosis)
Liver (cirrhosis)
Skin (Scleroderma, nephrogenic systemic fibrosis)
Joints (arthrofibrosis)
Kidney (renal fibrosis)
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7. Sample & Assay Technologies
Background: Fibrosis and diseases
Cardiac fibrosis
Can occur in response to left ventricular pressure-overload, as “reactive
interstitial fibrosis”, which leads to cardiac hypertrophy and necrosis
Alternatively, “replacement fibrosis” could occur in response to
myocardial infarction, inflammation, and myocyte death.
Systemic scleroderma - skin
Autoimmune disease of the skin and, in some cases, the internal organs
Arteriole endothelial and smooth muscle cell apoptosis, followed by
inflammation and fibrosis
Cause is unknown, but skin fibrosis can be treated
Liver cirrhosis
Develops as a result of chronic liver disease (alcoholic, fatty liver
disease, autoimmune disease, or hepatitis virus, etc.), or idiopathic
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8. Sample & Assay Technologies
Background: Fibrosis and diseases
Pulmonary fibrosis
Idiopathic (possibly linked to Surfactant protein C mutation) or as a result of injury or disease,
including:
Inhalation of particulates and gases
Smoking
Infections
Drugs (bleomycin, amiodarone, etc.)
Involvement by inflammatory mediators such as TNF, IL-beta, and IL-17, has been implicated, as well
as Th2 cytokines such as IL-13 and IL-4
Review, Integrating mechanisms of pulmonary fibrosis. JEM Vol. 208, No. 7
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9. Sample & Assay Technologies
Pulmonary fibrosis: role of T helper cells and macrophages
Review, Integrating mechanisms of pulmonary fibrosis. JEM Vol. 208, No. 7
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10. Sample & Assay Technologies
Researching the causes of fibrosis
Three recent case studies in scleroderma and cardiac fibrosis
Scleroderma and cytokines
SOCS3 and myocardial
infarction
Matrix metalloproteinases and
LV remodeling
Impaired IL-17 signaling
pathway contributes to the
increased collagen
expression in scleroderma
fibroblasts. Nakashima, T. et
al. (2012) Journal of
Immunology
Cardiac-specific deletion of
SOCS-3 prevents
development of left
ventricular remodeling after
acute myocardial infarction.
Oba, T. et al. (2012) Journal
of the American College of
Cardiology
Pressure overloaddependent membrane type Imatrix metalloproteinase
induction: relationship to LV
remodeling and fibrosis. Zile,
M.R. et al. (2012) American
Journal of Physiology, Heart
and Circulation Physiology
Used RT2 Profiler PCR Array
for Human Extracellular
Matrix and Adhesion
Molecules
Used RT2 Profiler PCR Array
for Mouse Common
Cytokines
Used a Custom RT2 Profiler
PCR Array.
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11. Sample & Assay Technologies
RT2 Profiler PCR Arrays
Pathway-focused gene expression profiling
-
84 (or 370) real-time PCR assays for genes related to specific pathways
Gene lists chosen by our experts – bioinformatics and text mining
Each assay is wet-lab tested for specificity and sensitivity
More than 140 pathways available, including many for fibrosis-related
processes
Integrated controls for normalization, reverse transcription, genomic DNA
contamination, and the PCR process, plus free data analysis tools
PCR Array Overview, Nov 15, 9:30am
https://www2.gotomeeting.com/register/263258378
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12. Sample & Assay Technologies
RT2 Profiler PCR Arrays for fibrosis and wound healing
Fibrosis (for Human, Rat, Mouse, Rabbit, and more)
http://sabiosciences.com/rt_pcr_product/HTML/PAHS-120Z.html
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13. Sample & Assay Technologies
RT2 Profiler PCR Arrays for fibrosis and wound healing
Wound healing (for Human, Mouse, Rat, Pig, Rabbit, and more)
http://sabiosciences.com/rt_pcr_product/HTML/PAHS-121Z.html
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14. Sample & Assay Technologies
RT2 Profiler PCR Arrays for fibrosis and wound healing
ECM & Adhesion Molecules (for Human, Mouse, Rat)
http://sabiosciences.com/rt_pcr_product/HTML/PAHS-013Z.html
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15. Sample & Assay Technologies
RT2 Profiler PCR Arrays for fibrosis and wound healing
Additional pathways available
Common Cytokines
Inflammatory Cytokines & Receptors
TGF-beta Signaling Pathway
TGF-beta Signaling Targets
Endothelial Cell Biology
Epithelial-to-Mesenchymal Transition
For complete list, see http://www.sabiosciences.com/ArrayList.php
Other species and custom array
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16. Sample & Assay Technologies
An application example using RT2 Profiler PCR Arrays
Cytokine expression changes in human PBMC on PMA/ionomycin
treatment.
Plot and Chart Format:
Heat Map
Scatter Plot
Volcano Plot
Clustergram
Multigroup Plot
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17. Sample & Assay Technologies
Researching the causes of fibrosis
3 recent applications in scleroderma, cardiac and lung fibrosis
Scleroderma and cytokines
SOCS3 and myocardial
infarction
Impaired IL-17 signaling pathway
contributes to the increased
collagen expression in
scleroderma fibroblasts.
Nakashima, T. et al. (2012)
Journal of Immunology
Pressure overload-dependent
membrane type I-matrix
Cardiac-specific deletion of
SOCS-3 prevents development of metalloproteinase induction:
relationship to LV remodeling and
left ventricular remodeling after
acute myocardial infarction. Oba, fibrosis. Zile, M.R. et al. (2012)
American Journal of Physiology,
T. et al. (2012) Journal of the
Heart and Circulation Physiology
American College of Cardiology
Used RT2 Profiler PCR Array for
Human Extracellular Matrix and
Adhesion Molecules
Matrix metalloproteinases and
LV remodeling
Used a Custom RT2 Profiler PCR
Array.
Used RT2 Profiler PCR Array for
Mouse Common Cytokines
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18. Sample & Assay Technologies
Application 1: Fibroblast collagen production and IL-17
Background and research question
Fibroblasts from SSc patients show intrinsic TGF-beta1
activation, and other cytokines are also implicated in disease
progression.
Previous studies yielded conflicting reports on the association of
IL-17 with SSc, and the authors sought to clarify its
involvement.
?
Are IL-17A&F and IL-17RA expressed differently
in SSc vs. healthy subjects?
Is IL-17 involved in regulating ECM during SSc?
Nakashima, T. et al. (2012) J. Immunol. 188, 3573.
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19. Sample & Assay Technologies
Application 1: Fibroblast collagen production and IL-17
Approach
Cytokines and receptors were measured in serum, fibroblast cultures,
and tissue samples by ELISA, immunoblotting, and
immunohistochemistry
An RT2 Profiler PCR Array for Human Extracellular Matrix and Adhesion
Molecules was used to profile ECM gene expression.
An RT2 miRNA PCR Array was used to profile miRNA expression
siRNA against TGF-beta1, Smad3, and IL-17RA were used to assess the
effects of TGF-beta signaling on IL-17 receptor expression and IL-17
signaling on miR-129-5p expression, respectively.
Nakashima, T. et al. (2012) J. Immunol. 188, 3573.
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20. Sample & Assay Technologies
Application 1: Fibroblast collagen production and IL-17
Major findings
IL-17A levels were higher in sera and involved skin of SSc patients, and
IL-17RA was lower at both the protein and mRNA level in cultured
fibroblasts. This was rescued by siRNA for TGF-beta1 or Smad
knockdown.
The RT2 Profiler PCR Array showed that IL-17A treatment caused
downregulation of pro-fibrotic CTGF. Alpha1(I) collagen expression
remained the same, but was lower by immunoblotting.
An RT2 miRNA PCR Array showed that miR-129-5p (among others) was
downregulated in SSc fibroblasts.
IL-17RA
CTGF
miR-129-5p
collagen
Nakashima, T. et al. (2012) J. Immunol. 188, 3573.
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21. Sample & Assay Technologies
Application 2: SOCS3 and myocardial infarction
Fibrosis research with RT2 Profiler PCR Arrays
Scleroderma and cytokines
SOCS3 and myocardial
infarction
Impaired IL-17 signaling pathway
contributes to the increased
collagen expression in
scleroderma fibroblasts.
Nakashima, T. et al. (2012)
Journal of Immunology
Pressure overload-dependent
membrane type I-matrix
Cardiac-specific deletion of
SOCS-3 prevents development of metalloproteinase induction:
relationship to :LV remodeling
left ventricular remodeling after
acute myocardial infarction. Oba, and fibrosis. Zile, M.R. et al.
(2012) American Journal of
T. et al. (2012) Journal of the
Physiology, Heart and Circulation
American College of Cardiology
Physiology
Used RT2 Profiler PCR Array for
Human Extracellular Matrix and
Adhesion Molecules
Used RT2 Profiler PCR Array for
Mouse Common Cytokines
Title, Location, Date
Matrix metalloproteinases and
LV remodeling
Used a Custom RT2 Profiler PCR
Array.
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22. Sample & Assay Technologies
Application 2: SOCS3 and myocardial infarction
Rationale and research question
Left ventricular remodeling after acute myocardial infarction
(AMI), including fibrosis, contributes to heart failure.
Previous work had shown that cytokines activating the
JAK/STAT pathways could prevent LV remodeling in animal
models after AMI.
? SOCS3 acts in a negative feedback loop induced
by JAK/STAT-activating cytokines – could inhibition of
SOCS3 prevent LV remodeling?
Oba, T. et al. (2012) Am. Coll. Cardiol. 59, 838.
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24. Sample & Assay Technologies
Application 2: SOCS3 and myocardial infarction
Approach
Made cardiac-specific SOCS3 knockout mice, induced AMI, and
observed LV remodeling in knockouts vs wild-types
Performed western blot analysis, TUNEL staining for apoptosis,
echocardiograph, and real-time PCR
Used Mouse Common Cytokines RT2 Profiler PCR Array to
profile cytokines in the system
Oba, T. et al. (2012) Am. Coll. Cardiol. 59, 838.
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25. Sample & Assay Technologies
Application 2: SOCS3 and LV remodeling
Major findings
Survival was enhanced in SOCS3 knockouts after AMI induced by
coronary ligation – 100% survived to 14 days, compared to 55% of
controls
LV remodeling was diminished in knockouts, as was apoptosis
RT2 Profiler PCR Array showed expression of multiple JAK-STATactivating cytokines following AMI, and many were diminished in SOCS3
knockouts (including G-CSF, IL-11, and IL-6)
Western blot showed greater activation of STAT3, AKT, and ERK
pathways in knockouts
Mallory-AZAN staining showed smaller fibrotic areas in knockout hearts,
and MMPs, TGF-beta2, and collagen showed lower expression as well
Conclusions
Cardiomyocyte SOCS3 may drive fibrosis development/LV
remodeling following AMI, and may be a useful therapeutic target.
Oba, T. et al. (2012) Am. Coll. Cardiol. 59, 838.
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26. Sample & Assay Technologies
Application 3: MT1-MMP, LV remodeling, and fibrosis
Fibrosis research with RT2 Profiler PCR Arrays
Scleroderma and cytokines
SOCS3 and myocardial
infarction
Matrix metalloproteinases and
LV remodeling
Impaired IL-17 signaling pathway
contributes to the increased
collagen expression in
scleroderma fibroblasts.
Nakashima, T. et al. (2012)
Journal of Immunology
Cardiac-specific deletion of
SOCS-3 prevents development of
left ventricular remodeling after
acute myocardial infarction. Oba,
T. et al. (2012) Journal of the
American College of Cardiology
Pressure overload-dependent
membrane type I-matrix
metalloproteinase induction:
relationship to :LV remodeling
and fibrosis. Zile, M.R. et al.
(2012) American Journal of
Physiology, Heart and Circulation
Physiology
Used RT2 Profiler PCR Array for
Human Extracellular Matrix and
Adhesion Molecules
Used RT2 Profiler PCR Array for
Mouse Common Cytokines
Used a Custom RT2 Profiler PCR
Array.
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27. Sample & Assay Technologies
Application 3: MT1-MMP, LV remodeling, and fibrosis
Research question
Myocardial fibrosis develops during chronic pressure-overload
(PO), which causes LV hypertrophy
Membrane type I MMP (MT1-MMP) is implicated in fibrosis
development, and its transcription is enhanced by mechanical
forces
? Could mechanical forces from chronic PO
increase MT1-MMP expression and fibrosis?
Zile, M. et al. (2012) Am. J. Physiol. Heart Circ. Physiol. 302, H1429
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28. Sample & Assay Technologies
Application 3: MT1-MMP, LV remodeling, and fibrosis
Approach
Developed an MT1-MMP promoter reporter mouse and used
transverse aortic constriction to model PO
Used a Custom RT2 Profiler PCR Array for MT1-MMP,
procollagen type I, CTGF, TGF-betaR1, and other profibrotic
genes in myocardial samples
Measured LV by echocardiography and collagen by light
microscopy
Isolated papillary muscles from reporter mice and subjected to
stimulation, then observed expression of MT1-MMP as well as
transcription factors
Zile, M. et al. (2012) Am. J. Physiol. Heart Circ. Physiol. 302, H1429
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29. Sample & Assay Technologies
Application 3: MT1-MMP, LV remodeling, and fibrosis
Major findings
PO led to LV hypertrophy and collagen volume fraction increase.
MT1-MMP protein abundance increased over the course of 4 weeks after
PO, and MT1-MMP promoter activity increased at 1 and 4 weeks, with a
dip at week 2.
The RT2 Profiler PCR Array showed increases in various profibrotic
genes, including the TGF-beta receptor, collagens, serine protease
inhibitors, LTBP, and CTGF, one week following PO.
Increases in mechanical load led to strong increases in MT1-MMP
expression in isolated papillary muscles, as well as expression of
transcription factors including NFkappB, RELA, and c-Fos.
Conclusions
Mechanical forces during PO may activate MT1-MMP transcription via
NFkappaB or c-Fos, exacerbating fibrosis through TGF-beta signaling. The
temporal associations in this study suggest that further research into MT1MMP as a driver of LV remodeling is warranted.
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30. Sample & Assay Technologies
RT2 Profiler PCR Arrays for fibrosis research
Interested in trying RT2 Profiler PCR Arrays?
Visit us at www.sabiosciences.com/rt_pcr_product to learn more!
Call
1-888-503-3187 for more information
Email:
support@SABiosciences.com (US & Canada)
sabio@qiagen.com (International customer)
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