scholarly journals QUANTITATIVE REAL-TIME PCR (RT-QPCR) COMPARING THE RELATIVE EXPRESSION LEVELS OF GENE TRANSCRIPTS INVOLVED IN A CRYPTIC THREE-WAY TRANSLOCATION T(9;11;19): AN ORIGINAL CASE OF AN INFANT WITH DISMAL PROGNOSIS ACUTE LYMPHOBLASTIC LEUKEMIA

2021 ◽  
Vol 43 ◽  
pp. S300
Author(s):  
GM Ferreira ◽  
RRC Matos ◽  
KC Monteso ◽  
MM Rocha ◽  
MT Bizarro ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Real Time ◽  
Real Time Pcr ◽  
Lymphoblastic Leukemia ◽  
Quantitative Real Time Pcr ◽  
Expression Levels ◽  
Relative Expression ◽  
Dismal Prognosis ◽  
Gene Transcripts ◽  
Original Case

scholarly journals Relative Expression Levels of SERCA in Panulirus argus and Homarus americanas tissues using Quantitative Real Time PCR

2006 ◽  
Vol 20 (5) ◽  
Author(s):  
Shyamala Arunachalam ◽  
Anita Mandal ◽  
Melisa Miller ◽  
Ella Meleshkevitch ◽  
Prabir K Mandal ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Panulirus Argus ◽  
Quantitative Real Time Pcr ◽  
Expression Levels ◽  
Relative Expression

scholarly journals Expression Analysis of Circulating miR-22, miR-122, miR-217 and miR-367 as Promising Biomarkers of Acute Lymphoblastic Leukemia

2021 ◽  
Author(s):  
Fatemeh hosseinpour-soleimani ◽  
Gholamreza Khamisipour ◽  
Zahra Derakhshan ◽  
Bahram Ahmadi
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Roc Analysis ◽  
Operating Characteristic ◽  
Lymphoblastic Leukemia ◽  
Area Under The Curve ◽  
Diagnostic Value ◽  
Healthy Individuals ◽  
Quantitative Real Time Pcr ◽  
Expression Levels

Abstract Background Currently, the role of serum-based biomarkers such as microRNAs in cancer diagnosis has been extensively established. This study aimed to determine expression levels of bioinformatically selected miRNAs and whether they can be used as biomarkers or a new therapeutic target in patients with Acute Lymphoblastic Leukemia (ALL). Materials and Methods The expression levels of serum miR-22, miR-122, miR-217, and miR-367 in 21 ALL patients and 21 healthy controls were measured using quantitative real-time PCR. The receiver operating characteristic (ROC) curve and the associated area under the curve (AUC) was used to assess candidate miRNAs' diagnostic value as a biomarker. Results The results showed that miR-217 was markedly decreased in patients with ALL compared to controls. Moreover, miR-22, miR-122, and miR-367 were found to be upregulated. Furthermore, ROC analysis showed that serum miR-217 and miR-367 could differentiate ALL patients from the healthy individuals, while miR-22 has approximate discriminatory power that requires further investigation. Conclusion Collectively, the results suggested that miR-217 may play a tumor suppressor role in ALL, whereas miR-22, miR-122, and miR-367 could function as an oncogene. Overall, miR-22, miR-217, and miR-367 could be considered possible biomarkers for the early diagnosis of ALL.

Discovery of Subtype-Specific Mirnas and New MiR-Genes In Pediatric Acute Lymphoblastic Leukemia

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3232-3232 ◽  
Author(s):  
Diana Schotte ◽  
Renee X de Menezes ◽  
Farhad Akbari Moqadam ◽  
Ellen Lange-Turenhout ◽  
Ronald W Stam ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Mirna Expression ◽  
Lymphoblastic Leukemia ◽  
Cell Type ◽  
Quantitative Real Time Pcr ◽  
Aberrant Expression ◽  
Stem Loop ◽  
Expression Levels ◽  
Genetic Subtypes ◽  
Pediatric All

Abstract Abstract 3232 MicroRNAs (miRNAs) are non-coding RNAs that regulate the activity of protein-coding genes and some miRNAs have been shown to be dysregulated in cancer. The function of miRNAs differs per cell type and prominent miRNAs in e.g. B-cell lymphoma may not be important in acute lymphoblastic leukemia (ALL). To identify which miRNAs may be relevant in pediatric ALL the expression level of 397 different miRNAs was measured in leukemic cells (>90% purity) of 81 pediatric ALL and 17 normal hematopoietic control samples by stem-loop reverse-transcriptase (RT) quantitative real-time PCR. Except for BCRABL1-positive and B-other ALL, all major subtypes including T-ALL, MLL-rearranged, TELAML1-positive, E2APBX1-positive and hyperdiploid (>50 chromosomes) ALL presented with unique miRNA signatures that differ from each other and from those in healthy hematopoietic cells. The expression levels highly varied amongst subtypes and, for example, differed by 4000-fold for miR-708 in T-ALL (P<0.0001) and 1700-fold for miR-383 in TELAML1-positive ALL (P<0.0001) compared to other precursor B-ALL cases. Some subtypes shared similarities in their miRNA expression signatures suggesting a common underlying biology; e.g. miR-196b was highly expressed in 9/12 MLL-rearranged and 14/22 T-ALL patients. In particular, all T-ALL cases carrying CALM-AF10, MLL-AF6, SET-NUP214 or an inversion of chromosome 7 expressed high levels of miR-196b similar to MLL-rearranged ALL. These genetic subtypes have all been functionally linked to upregulation of HOXA cluster genes. In correspondence, miR-196b expression levels were also highly correlated with HOXA (Rs>0.7, p<0.003), which was not seen for HOXB and HOXC cluster genes. High levels of miR-196b coincide with reduced methylation of the DNA at CpG islands directly upstream of miR-196b in MLL-rearranged cases compared to non-MLL precursor B-ALL and normal bone marrow cases, suggesting an epigenetic origin for miR-196b overexpression in these patients. High-throughput sequencing (Solexa technology) of small RNA libraries representing 7 subtypes of ALL and 3 normal hematopoietic tissue types followed by application of stringent computational miRNA precursor prediction algorithms resulted into the identification of 28 novel and 431 candidate novel miR-genes (besides 554 known miR-genes) for which the expression levels differed between genetic subtypes of pediatric ALL and normal hematopoietic cells. Stem-loop RT-quantitative real-time PCR confirmed aberrant expression levels of newly discovered miR-genes in a different set of patients. These data may point to leukemia-associated miRNAs for which the (aberrant) expression level and activity may be cell type (e.g. MLL-subtype) specific. These subtype-specific (known and novel) miRNAs may have stayed unrecognized when analyzed in a set of genetically unspecified ALL cases. In conclusion, genetic subtypes of ALL display unique miRNA expression signatures. Functional studies are in progress for these discriminative miRNAs since this may provide new insights into the biology of ALL subtypes. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Selection of reference genes for quantitative real-time PCR analysis in cucumber (Cucumis sativus L.), pumpkin (Cucurbita moschata Duch.) and cucumber–pumpkin grafted plants

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6536 ◽  
Author(s):  
Li Miao ◽  
Xing Qin ◽  
Lihong Gao ◽  
Qing Li ◽  
Shuzhen Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Reference Gene ◽  
Real Time Pcr ◽  
Reference Genes ◽  
Stable Expression ◽  
Quantitative Real Time Pcr ◽  
Graft Union ◽  
Expression Levels ◽  
Qrt Pcr

Background Quantitative real-time PCR (qRT-PCR) is a commonly used high-throughput technique to measure mRNA transcript levels. The accuracy of this evaluation of gene expression depends on the use of optimal reference genes. Cucumber–pumpkin grafted plants, made by grafting a cucumber scion onto pumpkin rootstock, are superior to either parent plant, as grafting conveys many advantages. However, although many reliable reference genes have been identified in both cucumber and pumpkin, none have been obtained for cucumber–pumpkin grafted plants. Methods In this work, 12 candidate reference genes, including eight traditional genes and four novel genes identified from our transcriptome data, were selected to assess their expression stability. Their expression levels in 25 samples, including three cucumber and three pumpkin samples from different organs, and 19 cucumber–pumpkin grafted samples from different organs, conditions, and varieties, were analyzed by qRT-PCR, and the stability of their expression was assessed by the comparative ΔCt method, geNorm, NormFinder, BestKeeper, and RefFinder. Results The results showed that the most suitable reference gene varied dependent on the organs, conditions, and varieties. CACS and 40SRPS8 were the most stable reference genes for all samples in our research. TIP41 and CACS showed the most stable expression in different cucumber organs, TIP41 and PP2A were the optimal reference genes in pumpkin organs, and CACS and 40SRPS8 were the most stable genes in all grafted cucumber samples. However, the optimal reference gene varied under different conditions. CACS and 40SRPS8 were the best combination of genes in different organs of cucumber–pumpkin grafted plants, TUA and RPL36Aa were the most stable in the graft union under cold stress, LEA26 and ARF showed the most stable expression in the graft union during the healing process, and TIP41 and PP2A were the most stable across different varieties of cucumber–pumpkin grafted plants. The use of LEA26, ARF and LEA26+ARF as reference genes were further verified by analyzing the expression levels of csaCYCD3;1, csaRUL, cmoRUL, and cmoPIN in the graft union at different time points after grafting. Discussion This work is the first report of appropriate reference genes in grafted cucumber plants and provides useful information for the study of gene expression and molecular mechanisms in cucumber–pumpkin grafted plants.

Gene-Related Strain Variation of Staphylococcus aureus for Homologous Resistance Response to Acid Stress

2014 ◽  
Vol 77 (10) ◽  
pp. 1794-1798 ◽  
Author(s):  
SOOMIN LEE ◽  
SOOYEON AHN ◽  
HEEYOUNG LEE ◽  
WON-IL KIM ◽  
HWANG-YONG KIM ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Real Time ◽  
Real Time Pcr ◽  
Acid Resistance ◽  
Transcriptional Analysis ◽  
Pcr Analysis ◽  
Quantitative Real Time Pcr ◽  
Resistance Response ◽  
Expression Levels ◽  
Acid Shock

This study investigated the effect of adaptation of Staphylococcus aureus strains to the acidic condition of tomato in response to environmental stresses, such as heat and acid. S. aureus ATCC 13565, ATCC 14458, ATCC 23235, ATCC 27664, and NCCP10826 habituated in tomato extract at 35°C for 24 h were inoculated in tryptic soy broth. The culture suspensions were then subjected to heat challenge or acid challenge at 60°C and pH 3.0, respectively, for 60 min. In addition, transcriptional analysis using quantitative real-time PCR was performed to evaluate the expression level of acid-shock genes, such as clpB, zwf, nuoF, and gnd, from five S. aureus strains after the acid habituation of strains in tomato at 35°C for 15 min and 60 min in comparison with that of the nonhabituated strains. In comparison with the nonhabituated strains, the five tomato-habituated S. aureus strains did not show cross protection to heat, but tomato-habituated S. aureus ATCC 23235 showed acid resistance. In quantitative real-time-PCR analysis, the relative expression levels of acid-shock genes (clpB, zwf, nuoF, and gnd) were increased the most in S. aureus ATCC 23235 after 60 min of tomato habituation, but there was little difference in the expression levels among the five S. aureus strains after 15 min of tomato habituation. These results indicate that the variation of acid resistance of S. aureus is related to the expression of acid-shock genes during acid habituation.

In Acute Promyelocytic Leukemia (APL) with Multiple Relapses Additional Prognostic Information Is Provided by FLT3 and Telomerase (hTERT) Quantitative Real-Time PCR.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4408-4408
Author(s):  
Paolo Bernasconi ◽  
Silvia Calatroni ◽  
Barbara Rocca ◽  
Paola Maria Cavigliano ◽  
Carlo Castagnola ◽  
...  
Keyword(s):  
Real Time ◽  
Induction Chemotherapy ◽  
Real Time Pcr ◽  
Clinical Relapse ◽  
Htert Expression ◽  
Quantitative Real Time Pcr ◽  
Molecular Remission ◽  
Expression Levels ◽  
Flt3 Expression ◽  
Multiple Relapses

Abstract The present study, carried out in two APL with multiple relapses, was aimed at determining 1) which correlation does exist between FLT3/hTERT expression levels and PML-RARA results; 2) whether high FLT3 and hTERT expression levels might be predictive of relapse; 3) whether FLT3 expression is better than FLT3 Internal Tandem Duplication (ITD) for evaluating disease outcome. Relative quantifications of FLT3/hTERT transcripts were performed by real-time PCR using SybrGreen I. For FLT3 calibration total RNA from a normal subject was used, for hTERT total RNA from K562 cells. In both cases the ΔΔCt method was used for quantification. On clinical diagnosis one patient with a WBC of 124.0x109/L and a PML-RARA fusion at PML BCR1 presented the FLT3/hTERT genes highly expressed. On qualitative PCR the patient also showed the ITD of FLT3. He was treated with the AIDA protocol and succeeded in achieving a haematological but not molecular remission. During CR FLT3/hTERT expression remained high and the ITD was never detected. Fourteen months later when on first clinical relapse FLT3 expression abruptly increased, the ITD reappeared, hTERT levels were still high. A re-induction chemotherapy induced a second haematological but not molecular remission lasting five months. FLT3 as well as hTERT expression levels became similar to those of the control, and FLT3 ITD disappeared. A progressive increase of FLT3 expression and an abrupt increase of hTERT expression preceded the second relapse which was accompanied by the reappearance of the ITD. After re-induction chemotherapy FLT3/hTERT expression dropped down to values of the control. A third CR was obtained but the patient remained PML/RARA and Flt3 ITD positive and soon after died of a CNS relapse. The other patient was treated in another Centre and came to our observation in haematological CR. At that time he was PML-RARA negative with high FLT3/hTERT expression. Eight months later he was still in clinical but not molecular CR having a PML-RARA fusion at BCR3, high FLT3/hTERT expression levels and presenting FLT3 ITD. One month later when clinical relapse occurred FLT3 expression levels were unchanged, hTERT expression dropped down to normal values and FLT3 ITD was still present. A re-induction chemotherapy induced a second CR with alternatively positive and negative PML-RARA results, high FLT3 and low hTERT expression levels. The patient underwent an allogeneic bone marrow transplant from an unrelated donor but five months later he relapsed for the second time with an abrupt rise of hTERT expression that preceded a quick increase of FLT3 expression. A third clinical but not molecular CR was achieved after chemotherapy, but the patient remained PML-RARA positive with a normal FLT3/hTERT expression. Two months later a rapid increase of hTERT expression preceded that of FLT3 and the occurrence of the third relapse. In conclusion i) increased FLT3 and hTERT levels during CR are associated with alternative positive/negative PML-RARA results on nested RT PCR and are always predictive of pending relapse; ii) on disease recurrence a marked elevation of hTERT expression often preceded that of FLT3; iii) quantitative real-time PCR of the FLT3 gene was more effective in predicting disease outcome than the ITD, this last being discovered only when FLT3 expression was already high.

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