scholarly journals OCT4B1 Regulates the Cellular Stress Response of Human Dental Pulp Cells with Inflammation

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Lu Liu ◽  
Rong Huang ◽  
Ruiqi Yang ◽  
Xi Wei
Keyword(s):  
In Situ Hybridization ◽  
Survival Rate ◽  
Western Blot ◽  
Real Time ◽  
Real Time Pcr ◽  
Dental Pulp ◽  
Immunofluorescence Staining ◽  
Dental Pulp Cells ◽  
Pulp Cells

Introduction. Infection and apoptosis are combined triggers for inflammation in dental tissues. Octamer-binding transcription factor 4-B1 (OCT4B1), a novel spliced variant of OCT4 family, could respond to the cellular stress and possess antiapoptotic property. However, its specific role in dental pulpitis remains unknown. Methods. To investigate the effect of OCT4B1 on inflammation of dental pulp cells (DPCs), its expression in inflamed dental pulp tissues and DPCs was examined by in situ hybridization, real-time PCR, and FISH assay. OCT4B1 overexpressed DPCs model was established, confirmed by western blot and immunofluorescence staining, and then stimulated with Lipopolysaccharide (LPS). Apoptotic rate was determined by Hoechst/PI staining and FACS. Cell survival rate was calculated by CCK8 assay. Results. In situ hybridization, real-time PCR, and FISH assay revealed that OCT4B1 was extensively expressed in inflamed dental pulp tissues and DPCs with LPS stimulation. Western blot and immunofluorescence staining showed the expression of OCT4B1 and OCT4B increased after OCT4B1 transfection. Hoechst/PI staining and FACS demonstrated that less red/blue fluorescence was detected and apoptotic percentage decreased (3.45%) after transfection. CCK8 demonstrated that the survival rate of pCDH-OCT4B1-flag cells increased. Conclusions. OCT4B1 plays an essential role in inflammation and apoptosis of DPCs. OCT4B might operate synergistically with OCT4B1 to reduce apoptosis.

scholarly journals XPC Promotes Pluripotency of Human Dental Pulp Cells through Regulation of Oct-4/Sox2/c-Myc

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Lu Liu ◽  
Zhengjun Peng ◽  
Zhezhen Xu ◽  
Xi Wei
Keyword(s):  
Western Blot ◽  
Real Time ◽  
Real Time Pcr ◽  
Dental Pulp ◽  
Dental Pulp Cells ◽  
Multilineage Differentiation ◽  
Human Dental Pulp Cells ◽  
Human Dental Pulp ◽  
Pulp Cells

Introduction. Xeroderma pigmentosum group C (XPC), essential component of multisubunit stem cell coactivator complex (SCC), functions as the critical factor modulating pluripotency and genome integrity through interaction with Oct-4/Sox2. However, its specific role in regulating pluripotency and multilineage differentiation of human dental pulp cells (DPCs) remains unknown. Methods. To elucidate the functional role XPC played in pluripotency and multilineage differentiation of DPCs, expressions of XPC in DPCs with long-term culture were examined by real-time PCR and western blot. DPCs were transfected with lentiviral-mediated human XPC gene; then transfection rate was investigated by real-time PCR and western blot. Cell cycle, apoptosis, proliferation, senescence, multilineage differentiation, and expression of Oct-4/Sox2/c-Myc in transfected DPCs were examined. Results. XPC, Oct-4, Sox2, and c-Myc were downregulated at P7 compared with P3 in DPCs with long-term culture. XPC genes were upregulated in DPCs at P2 after transfection and maintained high expression level at P3 and P7. Cell proliferation, PI value, and telomerase activity were enhanced, whereas apoptosis was suppressed in transfected DPCs. Oct-4/Sox2/c-Myc were significantly upregulated, and multilineage differentiation in DPCs with XPC overexpression was enhanced after transfection. Conclusions. XPC plays an essential role in the modulation of pluripotency and multilineage differentiation of DPCs through regulation of Oct-4/Sox2/c-Myc.

Quantification of an Eikelboom type 021N bulking event with fluorescence in situ hybridization and real-time PCR

2005 ◽  
Vol 68 (5) ◽  
pp. 695-704 ◽  
Author(s):  
Han Vervaeren ◽  
Kurt De Wilde ◽  
Jorg Matthys ◽  
Nico Boon ◽  
Lutgarde Raskin ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Real Time ◽  
Real Time Pcr

scholarly journals A 2,000-year-old specimen with intraerythrocytic Bartonella quintana

2020 ◽  
Author(s):  
R Barbieri ◽  
B-H-A Mai ◽  
T Chenal ◽  
M-L Bassi ◽  
D Gandia ◽  
...  
Keyword(s):  
Blood Cells ◽  
Dental Pulp ◽  
Glycophorin A ◽  
Dental Pulp Cells ◽  
Chain Reaction ◽  
Bartonella Quintana ◽  
Pulp Cells ◽  
Polymerase Chain ◽  
Indirect Tests

ABSTRACTPhotogrammetry and cascading microscopy investigations of dental pulp specimens collected from 2,000-year-old individuals buried in a Roman necropolis in Besançon, France, revealed unprecedented preserved tissular and cellular morphology. Photogrammetry yielded 3-D images of the smallest archaeological human remain ever recovered. Optical microscopy examinations after standard hematoxylin-phloxine-saffron staining and anti-glycophorin A immunohistochemistry exposed dental pulp cells, in addition erythrocytes were visualized by electron microscopy, which indicated that the ancient dental pulp trapped a blood drop. Fluorescence in situ hybridization applied on red blood cells revealed the louse-borne pathogen Bartonella quintana, a finding confirmed by polymerase chain reaction assays. Through paleohistology and paleocytology, we demonstrate that ancient dental pulp preserved intact blood cells at the time of the individual’s death, offering an unprecedented opportunity to engage in direct and indirect tests to diagnose pathogens in ancient buried individuals.

scholarly journals Histological Lesions and Replication Sites of PCV3 in Naturally Infected Pigs

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1520
Author(s):  
Elisa Rigo De Conti ◽  
Talita Pilar Resende ◽  
Lacey Marshall-Lund ◽  
Albert Rovira ◽  
Fabio Augusto Vannucci
Keyword(s):  
In Situ Hybridization ◽  
Lymph Nodes ◽  
Real Time ◽  
Real Time Pcr ◽  
Systemic Vasculitis ◽  
Tissue Type ◽  
Lymphoid Tissues ◽  
Associated Diseases ◽  
Spleen And Lymph Nodes

Porcine circovirus type 3 (PCV3) has been recently described as a potential cause of abortions and systemic vasculitis in pigs. Although the virus has been detected by real-time PCR in several porcine tissues from countries worldwide, PCV3-associated diseases have not been satisfactorily clarified. The objective of this study was to investigate the association between the presence of PCV3 mRNA detected by in situ hybridization (ISH) within histological lesions and PCV3 DNA detected by real-time PCR in naturally infected pigs. A total of 25 PCV3 PCR-positive cases were analyzed. Formalin-fixed tissues from these cases were evaluated for histologic lesions and for ISH-RNA positive signals for PCV3. The most frequent tissue type with histopathologic lesions was heart, 76.2%, with lymphoplasmacytic myocarditis and epicarditis as the most frequent lesions observed. Lymphoplasmacytic interstitial pneumonia was also a frequent finding, 47.6%. There were also lesions in kidney, liver, spleen and lymph nodes. PCV3-ISH-RNA positive signals were mostly observed in association with lymphoplasmacytic inflammatory infiltrate in various tissues, including arteries. Based on our results, the minimum set of specimens to be submitted for histopathology and mRNA in situ hybridization to confirm or exclude a diagnosis of PCV3 are heart, lung and lymphoid tissues (i.e., spleen and lymph nodes), especially for differential diagnosis related with PCV2-associated diseases.

The Recovery Effect of the Translationally Controlled Tumor Protein in 2-Hydroxy-Ethyl Methacrylate-Treated Human Dental Pulp Cells

2013 ◽  
Vol 747 ◽  
pp. 153-156
Author(s):  
Anchana Kongsaengkaeo ◽  
Wilaiwan Chotigeat ◽  
Ureporn Kedjarune-Leggat
Keyword(s):  
Real Time ◽  
Dental Pulp ◽  
Impedance Analysis ◽  
Dental Pulp Cells ◽  
Recovery Effect ◽  
Human Dental Pulp Cells ◽  
Translationally Controlled Tumor Protein ◽  
Ethyl Methacrylate ◽  
Human Dental Pulp ◽  
Pulp Cells

2-Hydroxy-ethyl methacrylate (HEMA) is a major monomer released from resin-base dental restorative materials. This study examined the recovery effect of the Translationally Controlled Tumor Protein (TCTP) in human dental pulp cells (HDPCs) after exposed to HEMA. TCTP from banana prawn (Penaeus merguiensis) was cloned and the protein was purified. A real-time cell analyzer was used to evaluate cell survival. The cell suspensions were seeded into an E-plate 96 at 8,000 cells/ well. After 24 hours, HDPCs were treated with 8 mM HEMA mixed in culture medium (alpha modified Eagles medium, α-MEM) for 1 hour before the medium supplemented with TCTP at 0, 100 ng/ml, 1 μg/ml, 10 μg/ml was replaced and left for 23 hours. After that, cells were fed with fresh medium for 72 hours. The cell indexes were monitored every 15 minutes and the 1-way analysis of variance (ANOVA) was used for statistical analysis. Real-time xCELLigence impedance analysis indicated that the cell indexes reached to 0 after treated with HEMA for 1 hour. TCTP at 1 μg/ml was significantly (P < 0.05) increased the cell index of HEME-treated HDPCs from 0 to 0.02 and 0.04 after TCTP exposure for 1 hour and 23 hours, respectively. It was suggested that TCTP has an ability to recover the cytotoxic effect of HEMA-treated pulp cells.

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