scholarly journals Glucocorticoid receptor expression and glucocorticoid therapeutic effect in nasal polyps

2010 ◽  
Vol 33 (3) ◽  
pp. 181 ◽  
Author(s):  
Peng Li ◽  
Yuan Li ◽  
Yong-Qi Li ◽  
Qin-Tai Yang ◽  
Ge-Hua Zhang
Keyword(s):  
Glucocorticoid Receptor ◽  
Mrna Expression ◽  
Therapeutic Effect ◽  
Nasal Mucosa ◽  
Nasal Polyps ◽  
Receptor Expression ◽  
Rt Pcr ◽  
Tissue Samples ◽  
Polymerase Chain ◽  
Sensitive Group

Purpose: To investigate the expression and quantity of glucocorticoid receptor-α and -β in polyp tissues taken from the patients treated were subsequently treated with topical glucocorticoid (GC). Methods: Eighty patients with nasal polyps were initially enrolled in the study. All polyp specimens were obtained prior to treatment. Patients then received daily topical GC spray treatment for one month. Polyp specimens were tested for glucocorticoid receptor (GR) GR-α and GR-β mRNA expression using fluorescent quantitative-reverse transcription-polymerase chain reaction (FQ-RT-PCR). Thirty healthy nasal mucosa tissue samples were tested at the same time. Results: Forty patients finished the study and were divided into two groups: GC-sensitive (n=26) and GC-insensitive (n=14), according to treatment results. GR-β mRNA expression in the nasal polyp tissues of the GC-insensitive group (5.72±0.58×102 copies/μg) was higher than that in the GC-sensitive group (4.82±0.28×102 copies/μg, P < 0.05) and in the normal nasal mucosa group (4.44±0.35×102 copies/μg, P < 0.01). There was also a difference in the relative expression of GR-α and GR-β between the GC-sensitive group (GR-α/GR-β= 829.42±67.36) and the GC-insensitive group (535.7±89) (P < 0.01). Conclusion: GR-β mRNA was highly expressed in patients with nasal polyps. Down- regulation of GR-α mRNA suggests the existence of glucocorticoid insensitivity. Expression of GR-β may plays an important role in the evaluation of the glucocorticoid therapeutic effect in patients with nasal polyps.

Glucocorticoid-induced glucocorticoid-receptor expression and promoter usage is not linked to glucocorticoid resistance in childhood ALL

Blood ◽  
2006 ◽  
Vol 108 (3) ◽  
pp. 1045-1049 ◽  
Author(s):  
Wim J. E. Tissing ◽  
Jules P. P. Meijerink ◽  
Bas Brinkhof ◽  
Mathilde J. C. Broekhuis ◽  
Renee X. Menezes ◽  
...  
Keyword(s):  
Glucocorticoid Receptor ◽  
Lymphoblastic Leukemia ◽  
Receptor Expression ◽  
Leukemic Cells ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Childhood All ◽  
Taqman Technology ◽  
Promoter Usage ◽  
Polymerase Chain

Abstract Glucocorticoid (GC) resistance is an adverse prognostic factor in childhood acute lymphoblastic leukemia (ALL), but little is known about causes of GC resistance. Up-regulation of the glucocorticoid receptor (GR) has been suggested as an essential step to the induction of apoptosis in leukemic cells. In this study we investigated whether baseline mRNA expression levels of the 5 different GR promoter transcripts (1A1, 1A2, 1A3, 1B, and 1C) or differences in the degree of regulation of the GR or GR promoter transcripts upon GC exposure are related to GC resistance. Therefore, mRNA levels of the 5 GR promoter transcripts and of the GR were measured by quantitative real-time reverse transcriptase–polymerase chain reaction (RT-PCR; Taqman) technology in primary ALL cells prior to and after 3, 8, and 24 hours of prednisolone exposure. GR expression is induced upon GC exposure in primary ALL patient samples, which is opposite to what is found in tissues in which GCs do not induce apoptosis. GC resistance in childhood ALL cannot be attributed to an inability of resistant cells to up-regulate the expression of the GR upon GC exposure, nor to differences in GR promoter usage (at baseline and upon GC exposure).

scholarly journals Expression Profile of IL-35 mRNA in Gingiva of Chronic Periodontitis and Aggressive Periodontitis Patients: A Semiquantitative RT-PCR Study

2013 ◽  
Vol 35 ◽  
pp. 819-823 ◽  
Author(s):  
Nagaraj B. Kalburgi ◽  
Akshay Muley ◽  
B. M. Shivaprasad ◽  
Arati C. Koregol
Keyword(s):  
Mrna Expression ◽  
Chronic Periodontitis ◽  
T Regulatory Cells ◽  
Periodontal Diseases ◽  
Aggressive Periodontitis ◽  
Gingival Tissue ◽  
Regulatory Cells ◽  
Rt Pcr ◽  
Tissue Samples ◽  
Polymerase Chain

Background. Proinflammatory and anti-inflammatory cytokines play a key role in the pathogenesis of periodontal diseases. Secretion of bioactive IL-35 has been described by T regulatory cells () and is required for their maximal suppressive activity. are involved in the modulation of local immune response in chronic periodontitis patients.Objective. Hence, the present study was aimed to investigate the expression of IL-35 mRNA in chronic periodontitis and aggressive periodontitis patients.Materials and Methods. The present study was carried out in 60 subjects, which included 20 chronic periodontitis patients, 20 aggressive periodontitis patients, and 20 periodontally healthy controls. IL-35 mRNA expression in gingival tissue samples of all subjects was semiquantitatively analyzed using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR).Results. The present study demonstrated the expression of IL-35 mRNA in gingival tissues of all the three groups. IL-35 mRNA expression was highest in chronic periodontitis subjects () as compared to the aggressive periodontitis group () and least seen in healthy patients ().Conclusion. The increased expression of IL-35 in chronic and aggressive periodontitis suggests its possible role in pathogenesis of periodontitis. Future studies done on large samples with intervention will strengthen our result.

scholarly journals The expression of endothelin type A and B receptors in the lateral wall of the mouse cochlea

2007 ◽  
Vol 12 (4) ◽  
Author(s):  
Yan Luo ◽  
Yuedi Tang ◽  
Qingjie Xia ◽  
Jin Liu
Keyword(s):  
Mrna Expression ◽  
Real Time ◽  
Lateral Wall ◽  
Type A ◽  
Mouse Tissue ◽  
Receptor Subtypes ◽  
Biological Functions ◽  
Rt Pcr ◽  
Tissue Samples ◽  
Polymerase Chain

AbstractEndothelin (ET), originally characterized as a vasoconstrictive peptide, has been found to have many different biological functions, including acting as a local hormonal regulator of pressure, fluid, ions and neurotransmitters in the inner ear. The objective of this study was to examine and quantify the mRNA expression of the endothelin type A and B receptors (ETAR and ETBR) in the strial vascularies (StV) and non-strial tissues (NSt) of the cochlear lateral wall using the real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) technique. The mouse tissue samples were harvested and RNA was extracted. RT was performed to obtain cDNA, and then the mRNA expression of each gene was measured via real-time PCR. We found that both receptor subtypes were expressed in the cochlear lateral wall, with a predominance of ETAR over ETBR. We showed that the mRNA expression of the two receptor subtypes was higher in the StV with a 1.8 times higher level of ETAR and an 8.1 times higher level of ETBR mRNAs than in the adjacent NSt of the lateral wall tissue. This study shows the existence and the quantity of ET receptor subtypes in the StV and NSt of the mouse cochlea. Our results suggest that an endothelin-mediated response via two different receptors, ETAR and ETBR, may play an important role in the physiological functions of the cochlear lateral wall by maintaining the homeostatic environment of the cochlea.

Gene expressions of TS and MDR-1 are predictive factors for chemosensitivity and survival in esophageal cancer with fluoropyrimidine-based chemotherapy

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 14120-14120
Author(s):  
K. Uchida ◽  
K. Hayashi ◽  
K. Kudo ◽  
H. Kuramochi ◽  
M. Yamamoto
Keyword(s):  
Esophageal Cancer ◽  
Mrna Expression ◽  
Overall Response Rate ◽  
Response Rate ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Gene Expressions ◽  
Polymerase Chain ◽  
Relative Mrna Expression ◽  
Mdr 1

14120 Background: Fluoropyrimidines are widely used in chemotherapy regimens for esophageal cancer. To test the hypotheses of whether the relative mRNA expression of the enzyme TS, DPD and MDR-1 were associated with response to in esophageal cancer. We investigated the associations reported between intratumoral TS, DPD and MDR-1 gene expressions and the response with 5-FU based chemotherapy for patients with esophageal cancer. Methods: Twenty six pts with esophageal cancer were treated with 5-FU based chemotherapy (14 pts were treated with chemotherapy only, and 12 pts were chemoradiotherapy). mRNA was isolated from frozen tumor specimens, and relative expression levels of each gene/β-actin were measured using a quantitative reverse transcription polymerase chain reaction (RT-PCR) (Taqman®) system. Results: The overall response rate was 50.0%. Each mRNA expression was detectable in 26 patients. TS expressions were significantly lower in the responding tumors compared to the non-responders respectively (P=0.009). There were no significant associations between MDR-1 expression and the response. By setting up a cut-off level for TS and MDR-1 of Median, combining the two gene expression two-dimensionally revealed a relationship with the response (P=0.037). Additionally, MDR-1 expressions were statistically significant predictors of prolonged survival (P=0.0084). Conclusions: These results suggest that intratumoral TS and MDR-1 expressions are prognostic factors for survival after 5-FU based chemotherapy in esophageal cancer. No significant financial relationships to disclose.

scholarly journals Differential Expression of Neurokinin-1 Receptor by Human Mucosal and Peripheral Lymphoid Cells

2000 ◽  
Vol 7 (3) ◽  
pp. 371-376 ◽  
Author(s):  
Triona Goode ◽  
Joe O'Connell ◽  
Wen-Zhe Ho ◽  
Gerald C. O'Sullivan ◽  
J. Kevin Collins ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Blood Lymphocytes ◽  
Lymphoid Cells ◽  
Receptor Expression ◽  
Rt Pcr ◽  
Peripheral Blood Mononuclear ◽  
Neurokinin 1 Receptor ◽  
Neurokinin 1

ABSTRACT Substance P (SP) has been implicated in peripheral and mucosal neuroimmunoregulation. However, confusion remains regarding immunocyte expression of the receptor for SP, neurokinin-1 receptor (NK-1R), and whether there is differential NK-1R expression in the mucosal versus the peripheral immune system. In the same assay systems, we examined the expression of NK-1R in human lamina propria mononuclear cells (LPMC), peripheral blood mononuclear cells (PBMC), peripheral blood lymphocytes (PBL), monocytes, and monocyte-derived macrophages (MDM). Using standard reverse transcription (RT)-PCR, mRNA expression of both the long and the short isoforms of the NK-1R was evident in LPMC but not in PBMC, PBL, monocytes, or MDM. However, by using nested RT-PCR NK-1R mRNA expression was detected in PBMC, PBL, monocytes, and MDM. This level of expression was found to represent one NK-1R mRNA transcript in >1,000 cells. In contrast, by using competitive RT-PCR we demonstrate that LPMC express a more biologically significant level of eight NK-1R mRNA transcripts per cell. Flow cytometric detection of NK-1R expression at the protein level was evident in LPMC but not in PBMC. These findings illustrate the extreme sensitivity of nested RT-PCR and the advantages of competitive RT-PCR in comparative studies of receptor expression in different cell populations. This study suggests that, under normal conditions, readily detectable expression of NK-1R in human mononuclear cells occurs at the mucosal level rather than in the peripheral circulation.

scholarly journals Aquaporin Expression in the Fetal Porcine Urinary Tract Changes During Gestation

2018 ◽  
pp. 283-292 ◽  
Author(s):  
L. K. JAKOBSEN ◽  
K. F. TRELBORG ◽  
P. S. KINGO ◽  
S. HØYER ◽  
K.-E. ANDERSSON ◽  
...  
Keyword(s):  
Urinary Tract ◽  
Mrna Expression ◽  
Western Blotting ◽  
Gestational Age ◽  
Renal Pelvis ◽  
Rt Pcr ◽  
Tissue Samples ◽  
Protein Levels ◽  
Water Handling ◽  
Aqp1 Expression

The expression of aquaporins (AQPs) in the fetal porcine urinary tract and its relation to gestational age has not been established. Tissue samples from the renal pelvis, ureter, bladder and urethra were obtained from porcine fetuses. Samples were examined by RT-PCR (AQPs 1-11), QPCR (AQPs positive on RT-PCR), and immunohistochemistry. Bladder samples were additionally examined by Western blotting. RNA was extracted from 76 tissue samples obtained from 19 fetuses. Gestational age was 60 (n=11) or 100 days (n=8). PCR showed that AQP1, 3, 9 and 11 mRNA was expressed in all locations. The expression of AQP3 increased significantly at all four locations with gestational age, whereas AQP11 significantly decreased. AQP1 expression increased in the ureter, bladder and urethra. AQP9 mRNA expression increased in the urethra and bladder, but decreased in the ureter. AQP5 was expressed only in the urethra. Immunohistochemistry showed AQP1 staining in sub-urothelial vessels at all locations. Western blotting analysis confirmed increased AQP1 protein levels in bladder samples during gestation. Expression levels of AQP1, 3, 5, 9 and 11 in the urinary tract change during gestation, and further studies are needed to provide insights into normal and pathophysiological water handling mechanisms in the fetus.

Mycoplasma pneumoniaeandChlamydophila pneumoniae: a comparative study in patients with nasal polyposis and healthy controls

2015 ◽  
Vol 129 (9) ◽  
pp. 865-869 ◽  
Author(s):  
D G Ioannidis ◽  
V A Lachanas ◽  
Z Florou ◽  
J G Bizakis ◽  
E Petinaki ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Chronic Rhinosinusitis ◽  
Nasal Polyps ◽  
Inferior Turbinate ◽  
Sinus Surgery ◽  
Control Group ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Polymerase Chain

AbstractIntroduction:The role played byMycoplasma pneumoniaeandChlamydophila pneumoniaein the pathogenesis of chronic rhinosinusitis with nasal polyps has been the object of ongoing debate. We used real-time polymerase chain reaction to investigate the prevalence of both microorganisms in the nasal tissue samples of patients and controls.Methods:We extracted DNA from nasal polyp samples obtained during functional endoscopic sinus surgery and the inferior turbinate samples of controls undergoing septoplasty. We used the highly sensitive real-time polymerase chain reaction to detect the presence ofM pneumoniaeandC pneumoniaeDNA.Results:Patients with chronic rhinosinusitis with nasal polyps consisted of 62 individuals (39 men; mean age 51 years); the control group consisted of 24 individuals (13 men; mean age 45 years). All samples from both groups were negative forM pneumoniaeandC pneumoniaeDNA.Conclusion:We have demonstrated that the likelihood ofM pneumoniaeandC pneumoniaeacting as an ongoing inflammatory stimulus in chronic rhinosinusitis with nasal polyps is slim.

Keratinocyte Growth Factor and its Receptor Messenger RNA Expression in Nasal Mucosa and Nasal Polyps

1998 ◽  
Vol 107 (10) ◽  
pp. 885-890 ◽  
Author(s):  
Toshio Ishibashi ◽  
Tadashi Tanaka ◽  
Shin-Ichi Ishimoto ◽  
Ken-Ichi Nibu ◽  
Kimitaka Kaga
Keyword(s):  
Growth Factor ◽  
Mrna Expression ◽  
Nasal Mucosa ◽  
Nasal Polyp ◽  
Messenger Rna ◽  
Nasal Polyps ◽  
Keratinocyte Growth Factor ◽  
Basic Fgf ◽  
Inflammatory Conditions

To examine the potential biologic role of fibroblast growth factors (FGFs) in nasal polyps and nasal mucosa during chronic inflammatory conditions, we investigated messenger RNA (mRNA) expression of three members of the FGF family — Acidic FGF, basic FGF, and keratinocyte growth factor (KGF) — in nasal polyp tissues, as well as in hyperplastic nasal mucosa. Using the sensitive method reverse transcription-polymerase chain reaction (RT-PCR), we demonstrated that of the examined FGFs, KGF had the most abundant mRNA expression in nasal polyps and nasal mucosa. We also found that significantly higher levels of KGF mRNA were expressed in nasal polyps than in nasal mucosa, whereas mRNA expression of acidic FGF and basic FGF was relatively low in these tissues. In addition, we showed that KGF receptor mRNA was present in most of the nasal mucosa; however, none or little was expressed in nasal polyps. These results suggest that KGF might play an important role in nasal epithelial proliferation and that excessive synthesis of KGF in nasal polyp stroma may contribute to hypertrophy of the nasal mucosa in patients with chronic sinusitis associated with nasal polyposis.

scholarly journals Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays

2000 ◽  
Vol 25 (2) ◽  
pp. 169-193 ◽  
Author(s):  
SA Bustin
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Housekeeping Gene ◽  
Absolute Quantification ◽  
Clear Understanding ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Quantitative Measurements ◽  
Polymerase Chain

The reverse transcription polymerase chain reaction (RT-PCR) is the most sensitive method for the detection of low-abundance mRNA, often obtained from limited tissue samples. However, it is a complex technique, there are substantial problems associated with its true sensitivity, reproducibility and specificity and, as a quantitative method, it suffers from the problems inherent in PCR. The recent introduction of fluorescence-based kinetic RT-PCR procedures significantly simplifies the process of producing reproducible quantification of mRNAs and promises to overcome these limitations. Nevertheless, their successful application depends on a clear understanding of the practical problems, and careful experimental design, application and validation remain essential for accurate quantitative measurements of transcription. This review discusses the technical aspects involved, contrasts conventional and kinetic RT-PCR methods for quantitating gene expression and compares the different kinetic RT-PCR systems. It illustrates the usefulness of these assays by demonstrating the significantly different levels of transcription between individuals of the housekeeping gene family, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH).

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