Quantitative PCR Analysis for Bcl-2/IgH in a Phase III Study of Yttrium-90 Ibritumomab Tiuxetan As Consolidation of First Remission in Patients With Follicular Lymphoma

2009 ◽  
Vol 27 (36) ◽  
pp. 6094-6100 ◽  
Author(s):  
Lindsey Goff ◽  
Karin Summers ◽  
Sameena Iqbal ◽  
Jens Kuhlmann ◽  
Michael Kunz ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Follicular Lymphoma ◽  
Quantitative Polymerase Chain Reaction ◽  
Phase Iii ◽  
Pcr Analysis ◽  
Control Group ◽  
Ibritumomab Tiuxetan ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Yttrium 90

Purpose The randomized First-Line Indolent Trial (FIT) was conducted in patients with advanced follicular lymphoma (FL), to evaluate the safety and efficacy of yttrium-90 (90Y) ibritumomab tiuxetan given as consolidation of complete or partial remission. This study of minimal residual disease was undertaken in parallel, to determine the rate of conversion from bcl-2 polymerase chain reaction (PCR) –detectable to –undetectable status and the corresponding effect on progression-free survival (PFS). Patients and Methods Blood samples from 414 patients (90Y-ibritumomab, n = 208; control, n = 206) were evaluated using real-time quantitative polymerase chain reaction (RQ-PCR); 186 were found to have the bcl-2 rearrangement and were thus eligible for inclusion in the RQ-PCR analysis. Results Overall, 90% of treated patients converted from bcl-2 PCR–detectable to –undetectable disease status, compared with 36% in the control group. Treatment significantly prolonged median PFS in patients converting to bcl-2 PCR-undetectable status (40.8 v 24.0 months in the control group; P < .01, hazard ratio [HR], 0.399). In patients who had bcl-2 PCR-detectable disease at random assignment, treatment significantly prolonged median PFS (38.4 v 8.2 months in the control group; P < .01, HR, 0.293). Conclusion Eradication of PCR-detectable disease occurred more frequently after treatment with 90Y-ibritumomab tiuxetan and was associated with prolongation of PFS.

scholarly journals Establishing and evaluation of a polymerase chain reaction for the detection of Echinococcus multilocularis in human tissue

2021 ◽  
Vol 15 (2) ◽  
pp. e0009155 ◽  
Author(s):  
Johannes Grimm ◽  
Julian Krickl ◽  
Annika Beck ◽  
Juliane Nell ◽  
Monika Bergmann ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Polymerase Chain Reaction ◽  
Echinococcus Multilocularis ◽  
Human Tissue ◽  
Pcr Analysis ◽  
Control Group ◽  
Chain Reaction ◽  
Robust Detection ◽  
Polymerase Chain ◽  
Laminated Layer

Background Alveolar echinococcosis (AE) is caused by metacestode larva of the tapeworm Echinococcus multilocularis. AE diagnostics currently rely on imaging techniques supported by serology, but unequivocal detection of AE is difficult. Although polymerase chain reaction (PCR)-based methods to detect tapeworm DNA in biopsies have been suggested for several species, no validated protocol adhering to accepted guidelines has so far been presented for AE diagnostics. We herein established a PCR protocol for metacestode biopsies and technically evaluated the method using isolated parasite DNA and cells, biopsies of clinically relevant material, and formalin fixed paraffin-embedded (FFPE) human tissue blocks. We compared the results with an immunochemical (IHC) approach using the monoclonal antibody Em2G11 specific for the antigen Em2 of E. mulitlocularis. Methodology/Principal findings Based on tapeworm 12S rDNA sequences we established and validated a PCR protocol for robust detection of as little as 50 parasite cells per specimen and report 127 cases of positive identification of Echinococcus species in samples from humans and animals. For further validation, we analyzed 45 liver, heart, brain, and soft tissue samples as well as cytological probes of aspirates of FFPE-material from 18 patients with clinically confirmed AE. Of each patient we analyzed (i) fully viable lesions with laminated layer; (ii) tissue with mAbEm2G11-positive small particles of E. multilocularis (spems); (iii) mAbEm2G11-negative tissue adjacent to the main lesion; and (iv) lymph node tissue with mAbEm2G11-positive spems. To identify the areas for the PCR-based approach, we performed IHC-staining with the monoclonal antibody Em2G11. Micro-dissected tissue of these areas was then used for PCR-analysis. 9 of 15 analyzed samples with viable E. multilocularis lesions with laminated layer were positive by PCR. Of this group, all samples preserved for less than 6 years (6/6) were tested positive. 11 of 15 samples of spems and 7 of 9 samples of the control group mAbEm2G11-negative tissue were negative by PCR. We further show that all probes from lymph nodes with spems are PCR negative. Conclusions/Significance We present a sensitive PCR method for the detection of E. multilocularis in human tissue, particularly in fresh biopsy material and tissue blocks stored for less than 5 years. While the diagnostic sensitivity of material containing only spems was higher using IHC, PCR detection was possible in IHC negative liver tissue and in patients with negative serology. Our results support the view that spems do not contain parasitic DNA or viable cells of the parasite. spems thus most probably do not directly contribute to metastasis formation during AE.

Decreased expression of fibulin-4 in aortic wall of aortic dissection

Vascular ◽  
2013 ◽  
Vol 22 (1) ◽  
pp. 35-41 ◽  
Author(s):  
P Huawei ◽  
C Qian ◽  
T Chuan ◽  
L Lei ◽  
W Liang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Aortic Dissection ◽  
Aortic Wall ◽  
Quantitative Polymerase Chain Reaction ◽  
Artery Bypass ◽  
Elastic Fiber ◽  
Elastic Fibers ◽  
Control Group ◽  
Chain Reaction ◽  
Polymerase Chain

In this research, we will examine the expression of Fibulin-4 in aortic wall to find out its role in aortic dissection development. The samples of aortic wall were obtained from 10 patients operated for acute ascending aortic dissection and five patients for chronic ascending aortic dissection. Another 15 pieces of samples from patients who had coronary artery bypass were as controls. The aortic samples were stained with aldehyde magenta dyeing to evaluate the arrangement of elastic fibers. The Fibulin-4 protein and mRNA expression were both determined by Western blot and realtime quantitative polymerase chain reaction. Compared with the control group, both in acute and chronic ascending aortic dissection, elastic fiber fragments increased and the expression of fibulin-4 protein significantly decreased ( P = 0.045 < 0.05). The level of fibulin-4 mRNA decreased in acute ascending aortic dissection ( P = 0.034 < 0.05), while it increased in chronic ascending aortic dissection ( P = 0.004 < 0.05). The increased amounts of elastic fiber fragments were negatively correlated with the expression of fibulin-4 mRNA in acute ascending aortic dissection. In conclusion, in aortic wall of ascending aortic dissection, the expression of fibulin-4 protein decreased and the expression of fibulin-4 mRNA was abnormal. Fibulin-4 may play an important role in the pathogenesis of aortic dissection.

scholarly journals Genetic Polymorphisms of Metabolic Enzyme Genes Associated with Leukocyte Mitochondrial DNA Copy Number in PAHs Exposure Workers

2020 ◽  
Author(s):  
Xinling Li ◽  
Xiaoran Duan ◽  
Hui Zhang ◽  
Mingcui Ding ◽  
Yanbin Wang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetic Polymorphisms ◽  
Peripheral Blood ◽  
Quantitative Polymerase Chain Reaction ◽  
Metabolic Enzymes ◽  
Control Group ◽  
Peripheral Blood Leukocytes ◽  
Chain Reaction ◽  
Blood Leukocytes ◽  
Polymerase Chain

Abstract Background: PAHs exposure had been reported to be a risk factor of mtDNAcn in our early study. However, the effect of metabolic enzymes’ genetic polymorphisms on mtDNAcn in PAHs-Exposure workers has not been fully evaluated.Methods: We investigated the effects of metabolic enzymes’ genetic polymorphisms on mtDNAcn among 544 coke oven workers and 238 office staffs. The mtDNAcn of peripheral blood leukocytes was measured using Real-time quantitative polymerase chain reaction method. Polymerase chain reaction and restriction fragment length was used to detect five polymorphisms in GSTT1, GSTM1, GSTP1 rs1695, CYP2E1 rs6413432, and CYP2E1 rs3813867.Results: The mtDNAcn in peripheral blood leukocytes was significantly lower in the exposure group than that in the control group (P < 0.001). The 1-OHPYR had an increasing trend with the genotypes AA→AG→GG of GSTP1 rs1695 in the control group. Generalized linear model indicated that the influencing factors of mtDNAcn were PAHs-exposure [b(95% CI) = -0.420 (-0.469, -0.372), P < 0.001], male [β(95% CI) = -0.058 (-0.103, -0.012), P = 0.013] ,and AA genotype for GSTP1 rs1695 [β(95% CI) = -0.051 (-0.095, -0.008), P = 0.020].Conclusions: The male was susceptibility to PAHs exposure. The AA genotype of GSTP1 rs1695 may influence the toxicity of PAHs and associated with the decreased of mtDNAcn.

scholarly journals Identification and Characterization of Osmoregulation Related MicroRNAs in Gills of Hybrid Tilapia Under Three Types of Osmotic Stress

2021 ◽  
Vol 12 ◽  
Author(s):  
Huanhuan Su ◽  
Jiajia Fan ◽  
Dongmei Ma ◽  
Huaping Zhu
Keyword(s):  
Polymerase Chain Reaction ◽  
Osmotic Stress ◽  
Expression Profiles ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Protein Translation ◽  
Control Group ◽  
Alkali Stress ◽  
Chain Reaction ◽  
Polymerase Chain

Researchers have increasingly suggested that microRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally regulate gene expression and protein translation in organs and respond to abiotic and biotic stressors. To understand the function of miRNAs in osmotic stress regulation of the gills of hybrid tilapia (Oreochromis mossambicus ♀ × Oreochromis urolepis hornorum ♂), high-throughput Illumina deep sequencing technology was used to investigate the expression profiles of miRNAs under salinity stress (S, 25‰), alkalinity stress (A, 4‰) and salinity–alkalinity stress (SA, S: 15‰, A: 4‰) challenges. The results showed that 31, 41, and 27 upregulated and 33, 42, and 40 downregulated miRNAs (P &lt; 0.05) were identified in the salt stress, alkali stress, and saline–alkali stress group, respectively, which were compared with those in the control group (C). Fourteen significantly differently expressed miRNAs were selected randomly and then validated by a quantitative polymerase chain reaction. On the basis of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis, genes related to osmoregulation and biosynthesis were enriched in the three types of osmotic stress. In addition, three miRNAs and three predicted target genes were chosen to conduct a quantitative polymerase chain reaction in the hybrid tilapia and its parents during 96-h osmotic stress. Differential expression patterns of miRNAs provided the basis for research data to further investigate the miRNA-modulating networks in osmoregulation of teleost.

The Role of Quantitative Polymerase Chain Reaction in the Management of Follicular Lymphoma Patients

2005 ◽  
Vol 91 (1) ◽  
pp. 59-66 ◽  
Author(s):  
Roberto A Perego ◽  
Roberto Cairoli ◽  
Giorgia Cornacchini ◽  
Cristina Bianchi ◽  
Matteo Corizzato ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Follicular Lymphoma ◽  
Quantitative Pcr ◽  
Quantitative Polymerase Chain Reaction ◽  
Prognostic Significance ◽  
Tumor Burden ◽  
Competitive Pcr ◽  
Molecular Remission ◽  
Chain Reaction ◽  
Polymerase Chain

Aims and background In order to increase the prognostic significance of polymerase chain reaction (PCR) data it has been suggested that quantitative PCR can be used to measure tumor burden. However, this option has not yet been definitely supported or refuted in patients with follicular lymphoma (FL). We decided to evaluate whether knowledge of the quantitative level of minimal residual disease and its variations can be of use in the management of FL patients. Methods We used qualitative and competitive PCR to study 11 patients with refractory or relapsed FL harboring the t(14;18) translocation who underwent autologous (nine patients) or allogeneic (two patients) stem cell transplantation (SCT). Competitive PCR was performed with a multiple competitor carrying specific sequences including Bcl2/lgH MBR and mcr, and the β-globin gene. Results After a median post-SCT follow-up of 44 months (range, 12-62), overall survival was 91% and disease-free survival 82%. The quantitative PCR data showed that: 1) effective chemotherapy before SCT substantially (1-2 log) reduced the tumor burden in the bone marrow (BM); 2) the increase in rearranged DNA detected in BM was associated with disease progression and relapse; 3) a PCR-negative autograft seemed to lead to lasting molecular remission even when it was performed in patients with a low level of BM infiltration before transplant; and 4) allo-SCT made and maintained the BM PCR negative even in the presence of a greater tumor burden before SCT. Six of the nine patients having CR after SCT (four auto and two allo) are in continuous molecular remission. Conclusions In FL patients qualitative and quantitative PCR may provide data that can be helpful for the prognostic evaluation of tumor progression and the early detection of impending relapse by highlighting biological features such as the quality of the infused material, the tumor burden at transplant, and the behavior of tumor cells after transplant.

scholarly journals High Expression MicroRNA-206 Inhibits the Growth of Tumor Cells in Human Malignant Fibrous Histiocytoma

2021 ◽  
Vol 9 ◽  
Author(s):  
Dejian Li ◽  
Kai Zhao ◽  
Ziwen Zhao ◽  
Bo Jiang ◽  
Xianxu Gong ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Tumor Cells ◽  
Malignant Fibrous Histiocytoma ◽  
Target Genes ◽  
Fibrous Histiocytoma ◽  
Quantitative Polymerase Chain Reaction ◽  
Control Group ◽  
Gene Probe ◽  
Chain Reaction ◽  
Polymerase Chain

Background: Malignant fibrous histiocytoma (MFH) is a common type of soft tissue sarcoma and a serious threat to human health. MFH often relapses locally after the curettage is related to the residual cancer stem cells (CSCs). Currently, the dysregulation of microRNA (miRNA) has been found to be closely related to the recurrence of CSCs. However, whether dysregulations of miRNAs exist in MFH, CSCs remained unknown.Methods: In this study, miRNAs in MFH CSCs and MFH common cells were examined by gene probe. Then, target genes and their functions involved in the signal pathway were predicted by the relevant database. Finally, the miRNAs’ target regulatory network was constructed. Furthermore, the miRNAs and target genes were identified by quantitative polymerase chain reaction, whereas miRNA analogs and antagonists were transfected in tumor cells to investigate cell proliferation ability further.Results: Results showed that a total of 47 miRNAs were found, including 16 that were upregulated and 31 that were downregulated. The screened differential miRNA showed a different expression in the cell resistant strains compared with the control group. Quantitative polymerase chain reaction analysis confirmed that the relative abundance of seven miRNAs and four target genes varied significantly. The encouraging issue is that we found Hsa-miR-206 significantly inhibited MFH proliferative activity.Conclusion: Hsa-miR-206 played a key role in regulating MFH CSC properties that might be a representative marker and target for the diagnosis and treatment of MFH in the future.

scholarly journals MicroRNA microarray analysis to detect biomarkers of aortic dissection from paraffin-embedded tissue samples

2020 ◽  
Vol 31 (2) ◽  
pp. 239-247
Author(s):  
Jun Ji ◽  
Qiong Xu ◽  
Xia He ◽  
Xiao-ling Chen ◽  
Jianan Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Real Time ◽  
Expression Profiles ◽  
Quantitative Polymerase Chain Reaction ◽  
Control Group ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract OBJECTIVES The aim of this study was to explore the differential expression profiles of microRNAs (miRNAs) in paraffin-embedded acute aortic dissection (AAD) tissues to find potential biomarkers for this disease. METHODS A total of 92 paraffin-embedded tissue specimens were collected from 92 patients with AAD who underwent surgical replacement. Among these specimens, 54 had partial normal aortic segments (smooth intima surface, non-atherosclerotic lesions) in proximal crevasse of aorta. Samples of these segments were taken 1 cm away from aortic lesions as the control group, after eliminating the tunica adventitia tissues. miRNA expression profiles were obtained by miRNA microarray analysis. Differentially expressed miRNAs were found by comparing the AAD group with the control group and were verified by fluorescence real-time quantitative polymerase chain reaction and by fluorescence in situ hybridization. RESULTS A total of 71 differentially expressed miRNAs were detected. Twenty-two were up-regulated and 49 were down-regulated. Four up-regulated miRNAs (hsa-miR-636, hsa-miR-142-3p, hsa-miR-425-3p, hsa-miR-191-3p) were selected for validation by real-time fluorescence quantitative polymerase chain reaction and fluorescence in situ hybridization. In the fluorescence real-time quantitative polymerase chain reaction analysis, only hsa-miR-636 showed a statistically significant difference in the AAD versus control comparison (3.3-fold, P = 0.012). The fluorescence in situ hybridization validation showed that the expression level of hsa-miR-636 was significantly increased in the AAD versus control comparison (P &lt; 0.001), with average optical densities of 61.29 ± 16.83 in the AAD group and 9.30 ± 3.98 in the control group. CONCLUSIONS Hsa-miR-636 is involved in the pathogenesis of AAD and may be a potential biomarker for this disease.

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