scholarly journals MicroRNA Analysis of Human Stroke Brain Tissue Resected during Decompressive Craniectomy/Stroke-Ectomy Surgery

Genes ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1860
Author(s):  
Andrew P. Carlson ◽  
William McKay ◽  
Jeremy S. Edwards ◽  
Radha Swaminathan ◽  
Karen S. SantaCruz ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Brain Tissue ◽  
Stroke Patient ◽  
Mirna Expression ◽  
Strong Association ◽  
Bioinformatic Analysis ◽  
Experimental Models ◽  
Aberrant Expression ◽  
Tissue Samples ◽  
Human Stroke

Background: Signaling pathways mediated by microRNAs (miRNAs) have been identified as one of the mechanisms that regulate stroke progression and recovery. Recent investigations using stroke patient blood and cerebrospinal fluid (CSF) demonstrated disease-specific alterations in miRNA expression. In this study, for the first time, we investigated miRNA expression signatures in freshly removed human stroke brain tissue. Methods: Human brain samples were obtained during craniectomy and brain tissue resection in severe stroke patients with life-threatening brain swelling. The tissue samples were subjected to histopathological and immunofluorescence microscopy evaluation, next generation miRNA sequencing (NGS), and bioinformatic analysis. Results: miRNA NGS analysis detected 34 miRNAs with significantly aberrant expression in stroke tissue, as compared to non-stroke samples. Of these miRNAs, 19 were previously identified in stroke patient blood and CSF, while dysregulation of 15 miRNAs was newly detected in this study. miRNA direct target gene analysis and bioinformatics approach demonstrated a strong association of the identified miRNAs with stroke-related biological processes and signaling pathways. Conclusions: Dysregulated miRNAs detected in our study could be regarded as potential candidates for biomarkers and/or targets for therapeutic intervention. The results described herein further our understanding of the molecular basis of stroke and provide valuable information for the future functional studies in the experimental models of stroke.

scholarly journals Signaling Pathways in Cervical Cancer Chemoresistance: Are microRNAs and Long-Noncoding RNAs the Main Culprits?

2020 ◽  
Author(s):  
Seyyed Reza Mousavi ◽  
Nima Hemmat ◽  
Hossein Bannazadeh Baghi ◽  
Afshin Derakhshani ◽  
Stefania Tommasi ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Drug Resistance ◽  
Signaling Pathways ◽  
Molecular Mechanisms ◽  
Bioinformatic Analysis ◽  
Aberrant Expression ◽  
Recurrent Cervical Cancer ◽  
Response To Chemotherapy ◽  
Wide Range ◽  
Non Coding Rnas

Cervical cancer is known as one of the most important cancers in women worldwide. Chemotherapy is a standard treatment for advanced/recurrent cervical cancer in which the prognosis of the disease is really poor and the 1-year survival chance in these patients is maximally 20%. However, resistance to anticancer drugs is a major problem in treating cancer. Cervical cancer stem cells are considered as a fundamental cause of chemo and radio-resistance and also relapse after primary successful treatment. Signaling pathways include a wide range of molecular mechanisms contribute to drug resistance. Recently, microRNAs (miRNAs) are announced as a group of molecular biomarkers involving in response to chemotherapy in cancer patients. As the miRNAs, there are some long non-coding RNAs (LncRNAs) which their aberrant expression is considered as a biomarker for monitoring chemoresistance. In this review, we summarized current reports about the involvement of signaling pathways during chemoresistance in cervical cancer. Then, genes that have been demonstrated their involvement during drug resistance in cervical cancer were tabulated. Further, miRNAs that have been reported as biomarkers during treatment are listed. By bioinformatic analysis, we predictedmiR-335-5p and miR-16-5p as the most potential biomarkers for monitoring resistance to chemotherapy. Finally, long non-coding RNAs that have been introduced in recent studies as novel biomarkers during the response to chemotherapy were mentioned.

scholarly journals Phosphoproteins Involved in the Inhibition of Apoptosis and in Cell Survival in the Leiomyoma

2019 ◽  
Vol 8 (5) ◽  
pp. 691 ◽  
Author(s):  
Blendi Ura ◽  
Lorenzo Monasta ◽  
Giorgio Arrigoni ◽  
Ilaria Battisti ◽  
Danilo Licastro ◽  
...  
Keyword(s):  
Cell Survival ◽  
Signaling Pathways ◽  
Protein Identification ◽  
Bioinformatic Analysis ◽  
Two Dimensional Gel Electrophoresis ◽  
Tissue Samples ◽  
New Class ◽  
Metal Affinity ◽  
Phosphorylated Proteins

Uterine leiomyomas are benign smooth muscle cell tumors originating from the myometrium. In this study we focus on leiomyoma and normal myometrium phosphoproteome, to identify differentially phosphorylated proteins involved in tumorigenic signaling pathways, and in anti-apoptotic processes and cell survival. We obtained paired tissue samples of seven leiomyomas and adjacent myometria and analyzed the phosphoproteome by two-dimensional gel electrophoresis (2-DE) combined with immobilized metal affinity chromatography (IMAC) and Pro-Q Diamond phosphoprotein gel stain. We used mass spectrometry for protein identification and Western blotting for 2-DE data validation. Quantities of 33 proteins enriched by the IMAC approach were significantly different in the leiomyoma if compared to the myometrium. Bioinformatic analysis revealed ten tumorigenic signaling pathways and four phosphoproteins involved in both the inhibition of apoptosis and cell survival. Our study highlights the involvement of the phosphoproteome in leiomyoma growth. Further studies are needed to understand the role of phosphorylation in leiomyoma. Our data shed light on mechanisms that still need to be ascertained, but could open the path to a new class of drugs that not only can block the growth, but could also lead to a significant reduction in tumor size.

scholarly journals IL-19 Promotes Nasal Mucosal Tissue Remodeling in Chronic Rhinosinusitis Patients By Upregulating Fibronectin and Collagen Ι Expressions in Fibroblasts Via NF-κB-Smad2/3 Signaling Pathways

2021 ◽  
Author(s):  
Hongwei Bao ◽  
Xia Li ◽  
Xiaoping Lai ◽  
Xiaoping Chen ◽  
Yue Li ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Chronic Rhinosinusitis ◽  
Nasal Polyps ◽  
Chronic Sinusitis ◽  
Interstitial Fibrosis ◽  
Tissue Remodeling ◽  
Control Group ◽  
Mucosal Tissue ◽  
Aberrant Expression ◽  
Tissue Samples

Abstract BackgroundChronic sinusitis without nasal polyps (CRSsNP), a subtype of chronic rhinosinusitis, is characterized by tissue remodeling, mainly by interstitial fibrosis. Fibroblasts are essential effectors of tissue remodeling, mainly expressing fibronectin (FN), collagen Ι (Col Ι), and other extracellular matrices. Our previous study found that IL-19 can promote the aberrant expression of extracellular matrix, and pre-experiments revealed that the Smad2/3 pathway plays a crucial role in tissue remodeling. However, the exact mechanism how IL-19 participants in tissue remodeling remains unclear. This study aimed to determine the roles of IL-19 in regulating the expression of FN and Col Ι, and the roles of Smad2/3 and NF-κB signaling pathways in fibroblasts.MethodsNasal mucosal tissue samples were collected from patients with chronic sinusitis with nasal polyps (CRSwNP), patients with CRSsNP, and controls to analyze the expression of IL-19, FN, and Col Ι by RT-qPCR, immunohistochemistry, immunofluorescence, or western blotting. Cultured primary human nasal mucosal fibroblasts were treated with human recombinant IL-19 with or without Smad2/3 and NF-κB pathway inhibitor or agonist. They were analyzed for Smad2/3 and NF-κB pathway activation, and FN and Col Ι expression in fibroblasts by western blot, immunohistochemistry, and immunofluorescence.ResultsIL-19 co-localized with FN and Col Ι in nasal mucosal tissues and the expression levels of FN and Col Ι were elevated in CRSsNP, compared with CRSwNP and the control group. IL-19 activated the Smad2/3 and NF-κB pathways and promoted FN and Col Ι production in fibroblasts. NF-κB functions as an upstream pathway of the Smad2/3 pathway. The inhibitors of Smad2/3 and NF-κB could significantly attenuate the expression of FN and Col Ι induced by IL-19. ConclusionIL-19 promotes FN and Col Ι expression in fibroblasts via the NF-κB-Smad2/3 signaling pathway, leading to fibrosis and collagen deposition in patients with CRSsNP.

scholarly journals Differential Notch and TGFβ Signaling in Primary Colorectal Tumors and Their Corresponding Metastases

2008 ◽  
Vol 30 (1) ◽  
pp. 1-11
Author(s):  
Liesbeth M. Veenendaal ◽  
Onno Kranenburg ◽  
Niels Smakman ◽  
Annemarie Klomp ◽  
Inne H. M. Borel Rinkes ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Lung Metastases ◽  
Tumor Development ◽  
Distant Metastases ◽  
Normal Mucosa ◽  
Normal Colonic Mucosa ◽  
Colorectal Tumors ◽  
Vimentin Expression ◽  
Aberrant Expression ◽  
Tissue Samples

Background: Loss of epithelial morphology and the acquisition of mesenchymal characteristics may contribute to metastasis formation during colorectal tumorigenesis. The Wnt, Notch and TGFβ signaling pathways control tissue homeostasis and tumor development in the gut. The relationship between the activity of these pathways and the expression of epithelial and mesenchymal markers was investigated in a series of primary colorectal tumors and their corresponding metastases.Methods: Tissue samples of primary colorectal tumors, normal colonic mucosa, and regional and systemic metastases were processed for immunohistochemistry in a tissue microarray format. The expression of mesenchymal (vimentin, fibronectin) and epithelial (E-cadherin) markers was related to markers of Wnt (β-catenin), Notch (HES1) and TGFβ (phospho-SMAD2) signalling. In addition, the KRAS mutation status was assessed.Results: When compared to normal mucosa, primary colorectal tumors showed a marked increase in the levels of cytoplasmic vimentin and nuclear β-catenin, phospho-SMAD2 and HES1. Increased vimentin expression correlated with the presence of oncogenic KRAS and with nuclear β-catenin. The corresponding liver, lymph node, brain and lung metastases did not express vimentin and displayed significantly lower levels of nuclear phospho-SMAD2 and HES1, while retaining nuclear β-catenin.Conclusions: Primary colorectal carcinomas display aberrant expression of vimentin, and have activated Notch and TGFβ signaling pathways. Surprisingly, many regional and distant metastases have lost nuclear HES1 and pSMAD2, suggesting that the activity of the Notch and TGFβ pathways is reduced in secondary colorectal tumors.

Effect of aberrant expression of miR-421, miR-29b_1_5p and miR-27b_5p on gastric adenoma development.

2018 ◽  
Vol 36 (4_suppl) ◽  
pp. 77-77
Author(s):  
Yu Jin Kim ◽  
Sang Woo Kim ◽  
Yong Chan Lee ◽  
Pumsoo Kim
Keyword(s):  
Gastric Cancer ◽  
Mirna Expression ◽  
High Grade Dysplasia ◽  
Low Grade ◽  
Gastric Adenoma ◽  
High Grade ◽  
Aberrant Expression ◽  
Tissue Samples ◽  
Altered Expression ◽  
Qrt Pcr

77 Background: Gastric cancer (GC) is the fourth leading cause of cancer-related death in the world. Some gastric adenomas may progress to adenocarcinoma in a short time. Several studies have examined adenomas the intermediate step of gastric carcinoma development. MicroRNAs have an important role in progression of gastric cancer but less is known about their role in premalignant adenomas. Changes in the expression pattern of miRNAs can be informative and highly significant in the gastric adenoma-carcinoma sequence progression as well. However, to date precancerous lesions (such as adenomas) have been investigated less frequently than cancers compared to healthy tissues. The aim of the present study was to perform a miRNA microarray analysis of normal, low-grade dysplasia, and high-grade dysplasia using fresh frozen tissues. Changes in miRNA expression patterns were also verified from samples obtained from the same patients using TaqMan MicroRNA Assays. Methods: Human tissue samples were obtained from nine patients who underwent endoscopic submucosal dissection (ESD) due to gastric adenoma or early gastric cancer at Severance Hospital of Yonsei University. Tumor and non-tumor mucosa tissue in each patient were taken just before ESD using biopsy forceps and frozen in the nitrogen tank. The miRNA expression profiles were analyzed by Affymetrix Gene-Chip miRNA 4.0 array (Homo sapiens). qRT-PCR was performed using TaqMan MicroRNA Assays (Applied Biosystems). Results: There were 28 (up-regulation) and 10 (down-regulation) miRNAs which showed significantly different expression between control group and low-/high-grade dysplasia group. miRNAs showing altered expression in normal and low-/high-grade dysplasia by microarrays were selected, and the expression tendencies were detected by real-time PCR validation. Three miRNAs (miR-421, miR-29b-1-5p, miR-27b-5p) were selected the continually upregulated from normal to low-/high-grade dysplasia based qRT-PCR in tissue samples. Conclusions: We identified new, less known miRNAs (miR-421, miR-29b-1-5p, miR-27b-5p), with altered expression in different grade dysplasia compare with normal samples.

scholarly journals Safety profile of the transcription factor EB (TFEB)-based gene therapy through intracranial injection in mice

2020 ◽  
Vol 11 (1) ◽  
pp. 241-250
Author(s):  
Zhenyu Li ◽  
Guangqian Ding ◽  
Yudi Wang ◽  
Zelong Zheng ◽  
Jianping Lv
Keyword(s):  
Gene Therapy ◽  
Transcription Factor ◽  
Neurodegenerative Diseases ◽  
Brain Tissue ◽  
Inflammatory Responses ◽  
Tissue Samples ◽  
Protein Levels ◽  
Virus Serotype ◽  
Transcription Factor Eb ◽  
The Brain

AbstractTranscription factor EB (TFEB)-based gene therapy is a promising therapeutic strategy in treating neurodegenerative diseases by promoting autophagy/lysosome-mediated degradation and clearance of misfolded proteins that contribute to the pathogenesis of these diseases. However, recent findings have shown that TFEB has proinflammatory properties, raising the safety concerns about its clinical application. To investigate whether TFEB induces significant inflammatory responses in the brain, male C57BL/6 mice were injected with phosphate-buffered saline (PBS), adeno-associated virus serotype 8 (AAV8) vectors overexpressing mouse TFEB (pAAV8-CMV-mTFEB), or AAV8 vectors expressing green fluorescent proteins (GFPs) in the barrel cortex. The brain tissue samples were collected at 2 months after injection. Western blotting and immunofluorescence staining showed that mTFEB protein levels were significantly increased in the brain tissue samples of mice injected with mTFEB-overexpressing vectors compared with those injected with PBS or GFP-overexpressing vectors. pAAV8-CMV-mTFEB injection resulted in significant elevations in the mRNA and protein levels of lysosomal biogenesis indicators in the brain tissue samples. No significant changes were observed in the expressions of GFAP, Iba1, and proinflammation mediators in the pAAV8-CMV-mTFEB-injected brain compared with those in the control groups. Collectively, our results suggest that AAV8 successfully mediates mTFEB overexpression in the mouse brain without inducing apparent local inflammation, supporting the safety of TFEB-based gene therapy in treating neurodegenerative diseases.

scholarly journals The m6A RNA methylation regulates oncogenic signaling pathways driving cell malignant transformation and carcinogenesis

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Mohammad Burhan Uddin ◽  
Zhishan Wang ◽  
Chengfeng Yang
Keyword(s):  
Signaling Pathways ◽  
Malignant Transformation ◽  
Noncoding Rna ◽  
Rna Modification ◽  
Oncogenic Signaling ◽  
Aberrant Expression ◽  
Rna Methylation ◽  
Rna Molecules ◽  
M6a Rna Methylation ◽  
Oncogenic Signaling Pathways

AbstractThe m6A RNA methylation is the most prevalent internal modification in mammalian mRNAs which plays critical biological roles by regulating vital cellular processes. Dysregulations of the m6A modification due to aberrant expression of its regulatory proteins are frequently observed in many pathological conditions, particularly in cancer. Normal cells undergo malignant transformation via activation or modulation of different oncogenic signaling pathways through complex mechanisms. Accumulating evidence showing regulation of oncogenic signaling pathways at the epitranscriptomic level has added an extra layer of the complexity. In particular, recent studies demonstrated that, in many types of cancers various oncogenic signaling pathways are modulated by the m6A modification in the target mRNAs as well as noncoding RNA transcripts. m6A modifications in these RNA molecules control their fate and metabolism by regulating their stability, translation or subcellular localizations. In this review we discussed recent exciting studies on oncogenic signaling pathways that are modulated by the m6A RNA modification and/or their regulators in cancer and provided perspectives for further studies. The regulation of oncogenic signaling pathways by the m6A modification and its regulators also render them as potential druggable targets for the treatment of cancer.

Evaluating the dynamics of brain tissue oxygenation using near-infrared spectroscopy on various experimental models

2019 ◽  
Vol 16 (11) ◽  
pp. 115602
Author(s):  
D M Kustov ◽  
A S Sharova ◽  
V I Makarov ◽  
A V Borodkin ◽  
T A Saveleva ◽  
...  
Keyword(s):  
Infrared Spectroscopy ◽  
Brain Tissue ◽  
Near Infrared Spectroscopy ◽  
Near Infrared ◽  
Tissue Oxygenation ◽  
Experimental Models ◽  
Brain Tissue Oxygenation

scholarly journals A Validated UPLC–MS-MS Assay for the Rapid Determination of Lorcaserin in Plasma and Brain Tissue Samples

2015 ◽  
Vol 40 (2) ◽  
pp. 133-139 ◽  
Author(s):  
Amal A. Bajrai ◽  
Essam Ezzeldin ◽  
Khalid A. Al-Rashood ◽  
Mohammad Raish ◽  
Muzaffar Iqbal
Keyword(s):  
Brain Tissue ◽  
Rapid Determination ◽  
Tissue Samples

Visualisation of GABAergic neurons and synapses in the rat brain using immunohistochemistry for two forms of glutamate decarboxylase

2021 ◽  
Vol 21 (2) ◽  
pp. 63-73
Author(s):  
Valeria A. Razenkova ◽  
Dmitrii E. Korzhevskii
Keyword(s):  
Brain Tissue ◽  
Glutamate Decarboxylase ◽  
Brain Structures ◽  
Gabaergic Neurons ◽  
Laboratory Animals ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Background Staining ◽  
The Central Nervous System ◽  
Rat Forebrain

BACKGROUND: Taking into account the importance of GABAergic brain system research and also the opportunity to achieve specific and accurate results in laboratory studies using immunohistochemical approaches, it seems important to have a reliable method of visualization GABA-synthesizing cells, their projections and synapses, for the morphofunctional analysis of GABAergic system both in normal conditions and in the experimental pathology. AIM: The aim of the study was to visualize analyze GABAergic neurons and synapses within rats brain using three different antibody types against glutamate decarboxylase and to identify the optimal conditions for reaction performing. MATERIALS AND METHODS: The study was performed on paraffin brain tissue sections of 5 adult Wistar rats. Immunohistochemical reactions using three antibody types against glutamate decarboxylase isoform 67 (GAD67) and glutamate decarboxylase isoform 65 (GAD65) were performed. Additional controls on C57/Bl6 mice and Chinchilla rabbits brain samples were also carried out. RESULTS: Antibodies used in the research made it possible to achieve high quality of GABAergic structures visualizing without increasing background staining. At the same time different antibody types are distinct in their efficacy to perform immunohistochemistry reaction on laboratory animal brain tissue samples. By performing additional controls, we discovered that there is necessary to adsorb secondary reagents immunoglobulins in order to eliminate nonspecific staining. It was found that GAD67 and GAD65 distribution in rat forebrain structures is different. It was stated that GAD67 immunohistochemistry most completely reveals GABAergic brain structures compared to GAD65 immunhistochemistry. The possibility of determining morphological features of GABAergic neurons and synaptic terminals, as well as performing quantitative analysis, was demonstrated. CONCLUSIONS: The approach proposed makes it possible to specifically visualize GABAergic structures of the central nervous system of different laboratory animals. This could be useful both in fundamental studies and in pathology research.

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