scholarly journals A d-2-hydroxyglutarate biosensor based on specific transcriptional regulator DhdR

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Dan Xiao ◽  
Wen Zhang ◽  
Xiaoting Guo ◽  
Yidong Liu ◽  
Chunxia Hu ◽  
...  
Keyword(s):  
Culture Medium ◽  
Transcriptional Regulator ◽  
High Specificity ◽  
Transcriptional Factor ◽  
Cell Culture Medium ◽  
Allosteric Effect ◽  
Specificity And Sensitivity ◽  
Lipopolysaccharide Biosynthesis ◽  
Homogeneous Assay

Abstractd-2-Hydroxyglutarate (d-2-HG) is a metabolite involved in many physiological metabolic processes. When d-2-HG is aberrantly accumulated due to mutations in isocitrate dehydrogenase or d-2-HG dehydrogenase, it functions in a pro-oncogenic manner and is thus considered a therapeutic target and biomarker in many cancers. In this study, DhdR from Achromobacter denitrificans NBRC 15125 is identified as an allosteric transcriptional factor that negatively regulates d-2-HG dehydrogenase expression and responds to the presence of d-2-HG. Based on the allosteric effect of DhdR, a d-2-HG biosensor is developed by combining DhdR with amplified luminescent proximity homogeneous assay (AlphaScreen) technology. The biosensor is able to detect d-2-HG in serum, urine, and cell culture medium with high specificity and sensitivity. Additionally, this biosensor is used to identify the role of d-2-HG metabolism in lipopolysaccharide biosynthesis of Pseudomonas aeruginosa, demonstrating its broad usages.

scholarly journals A D-2-hydroxyglutarate biosensor based on specific transcriptional regulator DhdR

2021 ◽  
Author(s):  
Dan Xiao ◽  
Wen Zhang ◽  
Xiaoting Guo ◽  
Yidong Liu ◽  
Chunxia Hu ◽  
...  
Keyword(s):  
Cell Culture ◽  
Transcriptional Regulator ◽  
High Specificity ◽  
Allosteric Effect ◽  
Transcription Regulator ◽  
Specificity And Sensitivity ◽  
Lipopolysaccharide Biosynthesis ◽  
Domains Of Life ◽  
Homogeneous Assay

AbstractD-2-Hydroxyglutarate (D-2-HG) is a metabolite in many physiological metabolic processes. When D-2-HG is aberrantly accumulated due to mutations in isocitrate dehydrogenases or D-2-HG dehydrogenase, it functions in a pro-oncogenic manner and is thus considered a therapeutic target and biomarker in many cancers. In this study, DhdR from Achromobacter denitrificans NBRC 15125 was identified as an allosteric transcription factor that negatively regulates D-2-HG dehydrogenase expression and responds to presence of D-2-HG. It is the first known transcription regulator specifically responding to D-2-HG across all domains of life. Based on the allosteric effect of DhdR, a D-2-HG biosensor was developed by combining DhdR with amplified luminescent proximity homogeneous assay technology. The biosensor was able to detect D-2-HG in serum, urine, and cell culture with high specificity and sensitivity. Additionally, this biosensor was also successfully used to identify the role of D-2-HG metabolism in lipopolysaccharide biosynthesis of Pseudomonas aeruginosa, demonstrating its broad usages.

scholarly journals Cellular discrimination using in vitro Raman micro spectroscopy: the role of the nucleolus

The Analyst ◽  
2015 ◽  
Vol 140 (17) ◽  
pp. 5908-5919 ◽  
Author(s):  
Z. Farhane ◽  
F. Bonnier ◽  
A. Casey ◽  
A. Maguire ◽  
L. O'Neill ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
High Specificity ◽  
Cancer Cell Lines ◽  
Cellular Organelle ◽  
Specificity And Sensitivity

Raman micro spectroscopy is employed to discriminate between cell lines. Results show the importance of the nuclear sub-cellular organelle, the nucleoli, to differentiate between cancer cell lines with high specificity and sensitivity.

scholarly journals The Role of Breast Cancer Stem Cell-Related Biomarkers as Prognostic Factors

Diagnostics ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 721
Author(s):  
Clarence Ching Huat Ko ◽  
Wai Kit Chia ◽  
Gayathri Thevi Selvarajah ◽  
Yoke Kqueen Cheah ◽  
Yin Ping Wong ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Stem Cell ◽  
Cancer Stem Cell ◽  
High Specificity ◽  
Breast Cancer Diagnosis ◽  
Breast Cancer Stem Cell ◽  
Prognostic Indicators ◽  
Specificity And Sensitivity

Breast cancer is one of the leading causes of cancer-related deaths in women worldwide, and its incidence is on the rise. A small fraction of cancer stem cells was identified within the tumour bulk, which are regarded as cancer-initiating cells, possess self-renewal and propagation potential, and a key driver for tumour heterogeneity and disease progression. Cancer heterogeneity reduces the overall efficacy of chemotherapy and contributes to treatment failure and relapse. The cell-surface and subcellular biomarkers related to breast cancer stem cell (BCSC) phenotypes are increasingly being recognised. These biomarkers are useful for the isolation of BCSCs and can serve as potential therapeutic targets and prognostic tools to monitor treatment responses. Recently, the role of noncoding microRNAs (miRNAs) has extensively been explored as novel biomarker molecules for breast cancer diagnosis and prognosis with high specificity and sensitivity. An in-depth understanding of the biological roles of miRNA in breast carcinogenesis provides insights into the pathways of cancer development and its utility for disease prognostication. This review gives an overview of stem cells, highlights the biomarkers expressed in BCSCs and describes their potential role as prognostic indicators.

scholarly journals von Willebrand Factor and Oxidative Stress Parameters in Acute Coronary Syndromes

2011 ◽  
Vol 2011 ◽  
pp. 1-11 ◽  
Author(s):  
Zoran Koprivica ◽  
Dusica Djordjevic ◽  
Milena Vuletic ◽  
Vladimir Zivkovic ◽  
Nevena Barudzic ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Von Willebrand Factor ◽  
Acute Coronary Syndromes ◽  
High Specificity ◽  
Independent Effect ◽  
Specificity And Sensitivity ◽  
Coronary Syndromes ◽  
Von Willebrand ◽  
Willebrand Factor

Considering the role of von Willebrand factor (vWf) in hemostasis, and the role of oxidative stress in the development of endothelial dysfunction and atherosclerotic disease, the aim of our study was to investigate the relationship between vWf, parameters of oxidative stress and different types of acute coronary syndromes (ACS). Levels of vWf activity (vWfAct), vWf antigen (vWfAg), nitric oxide (estimated through nitrites–NO2 −), superoxide anion radical (O2 −), hydrogen peroxide (H2O2), index of lipid peroxidation (estimated through thiobarbituric acid reactive substances–TBARS), superoxide dismutase (SOD) and catalase (CAT) activity of 115 patients were compared with those of 40 healthy controls. ACS patients had significantly higher vWfAct and vWfAg levels, as well as TBARS levels, while their levels of NO2 −, H2O2, SOD and CAT activities were lower than controls'. vWfAg showed high specificity and sensitivity as a test to reveal healthy or diseased subjects. Multivariant logistic regression marked only vWfAg and TBARS as parameters that were under independent effect of ACS type. The results of our study support the implementation of vWf in clinical rutine and into therapeutic targets, and suggest that ACS patients are in need of antioxidant supplementation to improve their impaired antioxidant defence.

scholarly journals Diagnostic and Prognostic Value of Three microRNAs in Environmental Asbestiform Fibers-Associated Malignant Mesothelioma

2021 ◽  
Vol 11 (11) ◽  
pp. 1205
Author(s):  
Veronica Filetti ◽  
Carla Loreto ◽  
Luca Falzone ◽  
Claudia Lombardo ◽  
Emanuele Cannizzaro ◽  
...  
Keyword(s):  
Malignant Mesothelioma ◽  
Asbestos Exposure ◽  
High Specificity ◽  
Digital Pcr ◽  
Diagnostic Value ◽  
Diagnostic Significance ◽  
Diagnostic Potential ◽  
Specificity And Sensitivity ◽  
Diagnostic And Prognostic Value

Fluoro-edenite (FE) is an asbestiform fiber identified in Biancavilla (Sicily, Italy). Environmental exposure to FE has been associated with a higher incidence of malignant mesothelioma (MM). The present study aimed to validate the predicted diagnostic significance of hsa-miR-323a-3p, hsa-miR-101-3p, and hsa-miR-20b-5p on a subset of MM patients exposed to FE and matched with healthy controls. For this purpose, MM tissues vs. nonmalignant pleura tissues were analyzed through droplet digital PCR (ddPCR) to evaluate differences in the expression levels of the selected miRNAs and their MM diagnostic potential. In addition, further computational analysis has been performed to establish the correlation of these miRNAs with the available online asbestos exposure data and clinic-pathological parameters to verify the potential role of these miRNAs as prognostic tools. ddPCR results showed that the three analyzed miRNAs were significantly down-regulated in MM cases vs. controls. Receiver operating characteristic (ROC) analysis revealed high specificity and sensitivity rates for both hsa-miR-323a-3p and hsa-miR-20b-5p, which thus acquire a diagnostic value for MM. In silico results showed a potential prognostic role of hsa-miR-101-3p due to a significant association of its higher expression and increased overall survival (OS) of MM patients.

scholarly journals Role of Fnac In Evaluation of Parotid Gland Swelling

2019 ◽  
Vol 17 (1) ◽  
pp. 30-34
Author(s):  
Krisna Rani Majumder ◽  
Syed Serajul Karim ◽  
Md Monjurul Alam
Keyword(s):  
Parotid Gland ◽  
High Specificity ◽  
Needle Aspiration ◽  
Low Grade ◽  
Major Salivary Gland ◽  
Small Series ◽  
Specificity And Sensitivity ◽  
Parotid Neoplasm ◽  
Benign Tumours

Objectives: To correlate the findings of FNAC with final histological report in parotid gland swelling and to find out the histological types of parotid neoplasm. Methods: An attempt has been made to find out age and sex distribution, FNAC findings and histological type of parotid gland neoplasm. A total 30 cases were studied from the inpatient departments of General Surgery and Otolaryngology, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka since July, 2011 to April, 2013. Result: Regarding investigations in this series Fine Needle Aspiration Cytology (FNAC) was done in all 30(100%). In this series out of 30 cases of parotid neoplasm 23 (76.66%) cases were benign and rest 07 (23.33%) were malignant. Among the benign tumours commonest was pleomorphic adenoma 20 (66.66%) and regarding malignancy Mucoepidermoid Carcinoma was most common 3 (10%) then adenoid cystic carcinoma 2 (6.66%). Out of 7, 3 were stage 3(42.85%) and 3 were stage 4(42.85%). In histological grading 3 were low grade (42.85%) and 4 were high grade (57 .14%). All the patient underwent partotidectomy. Few were treated by radical parotidectomy plus radiotherapy. Early diagnosis and treatment of major salivary gland tumour is likely to lead a fare outcome. Conclusion: FNAC is a useful preoperative diagnostic tool for parotid gland swelling with high specificity and sensitivity. Though it is a small series to comment on the whole spectrum of the parotid gland swelling in Bangladesh, yet this study has been designed to see the role of FNAC in neoplastic parotid lesion which will be beneficial for proper management. Journal of Surgical Sciences (2013) Vol. 17 (1) : 30-34

scholarly journals Loss of the ClpXP Protease Leads to Decreased Resistance to Cell-Envelope Targeting Antimicrobials in Bacillus anthracis Sterne

2021 ◽  
Vol 12 ◽  
Author(s):  
Lang Zou ◽  
Christopher R. Evans ◽  
Vuong D. Do ◽  
Quinn P. Losefsky ◽  
Diem Q. Ngo ◽  
...  
Keyword(s):  
Cell Culture ◽  
Bacillus Anthracis ◽  
Culture Medium ◽  
Cell Envelope ◽  
Mammalian Cell Culture ◽  
Antibiotic Sensitivity ◽  
Cell Culture Medium ◽  
Intrinsic Resistance ◽  
Cellular Density

The ClpX ATPase is critical for resistance to cell envelope targeting antibiotics in Bacillus anthracis, however, it is unclear whether this is due to its function as an independent chaperone or as part of the ClpXP protease. In this study, we demonstrate that antibiotic resistance is due to formation of the ClpXP protease through construction of a ClpX complementation plasmid that is unable to interact with ClpP. Additionally, we genetically disrupted both clpP genes, clpP1 and clpP2, found in B. anthracis Sterne and find that the loss of either increases susceptibility to cell envelope targeting antimicrobials, although neither has as strong of a phenotype as loss of clpX and neither clpP gene is essential for virulence in a G. mellonella model of infection. Lastly, we looked at changes to cell envelope morphology that could contribute to increased antibiotic sensitivity. We find no difference in cell charge or cell lysis, although we do see increased hydrophobicity in the ΔclpX strain, decreased cellular density and slightly thinner cells walls. We also see significant cell division defects in ΔclpX, although only when cells are grown in the mammalian cell culture medium, RPMI. We conclude that the intrinsic resistance of B. anthracis to cell wall active antimicrobials is dependent on formation of the ClpXP protease and that this could be due, at least in part, to the role of ClpX in regulating cell envelope morphology.

scholarly journals Direct and Indirect Control of Late Sporulation Genes by GerR of Bacillus subtilis

2010 ◽  
Vol 192 (13) ◽  
pp. 3406-3413 ◽  
Author(s):  
Giuseppina Cangiano ◽  
Antonio Mazzone ◽  
Loredana Baccigalupi ◽  
Rachele Isticato ◽  
Patrick Eichenberger ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Rna Polymerase ◽  
Chromatin Immunoprecipitation ◽  
Transcriptional Regulator ◽  
Transcriptional Factor ◽  
Negative Regulator ◽  
Promoter Regions ◽  
Indirect Control ◽  
Sporulation Genes

ABSTRACT GerR is a sporulation-specific transcriptional factor of Bacillus subtilis that has been identified as a negative regulator of genes transcribed by σE-containing RNA polymerase and as a positive effector of the expression of three late sporulation genes. Here we confirmed that gerR transcription is dependent on σE-containing RNA polymerase but also observed that it requires the transcriptional regulator SpoIIID. The study of the role of GerR in regulating the expression of several late sporulation genes allowed us to observe that its effect is strongly positive on spoVIF, cotC, and cotG, weakly positive on cotB, and negative on cotU. The results of chromatin immunoprecipitation (ChIP) experiments indicated that GerR binds to the promoter regions of some, but not all, of the GerR-controlled genes, leading us to propose that GerR controls late sporulation genes in two ways: (i) directly, by acting on the transcription of cotB, cotU and spoVIF; and (ii) indirectly, through the activation of SpoVIF, which stabilizes the transcriptional activator GerE and consequently induces the expression of the GerE-dependent genes cotC and cotG.

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