scholarly journals Lead and cadmium influences on the metallothionein expression level in Lymnaea stagnalis L. adults

1970 ◽  
pp. 281-285
Author(s):  
S. E. Dromashko ◽  
S. N. Shevtsova ◽  
A. S. Babenko
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Lymnaea Stagnalis ◽  
Test System ◽  
Expression Level ◽  
Pond Snail ◽  
Cadmium Ions ◽  
Lead And Cadmium ◽  
Expression Evaluation

Aim. The article deals with the assessment of some heavy metals effects on metallothionein expression in such a test system as the great pond snail L. stagnalis. Methods. There are methods of mollusk specimen cultivation and gene expression level estimation (PCR in real-time mode), as well as data analysis. Results. A significant increase in MT gene expression was observed in both young (4.5 months) and older (7 months) mollusks on the 7th day of exposure to 0.003 mg/L of lead ions as well as significant increase in MT gene expression was observed in older mollusks on the 7th day of exposure to 0.0001–0.001 mg/L of cadmium ions. Conclusions. The expediency of L. stagnalis adults using at the age of 3 to 5 months with other test systems for biotesting liquid metal-containing waste based on the gene expression evaluation in a short (no more than 7 day) experiment using the Real Time PCR has been shown.Keywords: Lymnaea stagnalis, lead/cadmium, metallothioneins, gene expression, real time PCR.

Real-Time PCR Gene Expression Profiling of Microarray Gene Signatures in Acute Leukemia.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4401-4401
Author(s):  
Ebrahim Sakhinia ◽  
Mahboubeh Farahangpour ◽  
John A. Liu Yin ◽  
Gerard Brady ◽  
Judith A. Hoyland ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Cyclin D3 ◽  
Expression Level ◽  
Tumor Type ◽  
Gene Signatures ◽  
Significant Difference ◽  
The Mean ◽  
Microarray Gene

Abstract Cancer subtype discovery and classification using microarray gene signatures has the potential to transform pathological diagnosis but measurement of indicator genes in routine practice remains difficult. We tested use of real-time PCR measurement of indicator genes for AML and ALL (Golub et al, Science, 1999) as a method for validation and application of microarray gene signatures. Mononuclear cells (MC) were isolated from whole bone marrow (BM) aspirates by density gradient centrifugation and sorted into unselected (total), CD34+ve and CD34-ve fractions. The mRNA in each fraction was globally amplified using a PolyA PCR method. We measured the expression profile of the 17 top ranked genes (cystatin C, leptin receptor, fumarylacetoacetate, CD33, HoxA9, adipsin, proteoglycan 1, LTC4 synthase, LYN, C-myb, MB-1, cyclin D3, SNF2, RbAp48, proteasome iota, HkrT-1 and E2A) from Golub et al (1999) by real-time PCR. All values were calibrated against control standards and normalized to the mean of three housekeeping genes (IF2-beta, GAPDH and human ribosomal protein S9). Data for all 17 genes were obtained for 4 (ALL), 26 (AML), 12 (AML remission) and 9 (morphologically normal) BM samples, each fractionated into three fractions (total MC, CD34+ve MC & CD34−ve MC). There was no significant difference in the mean of three housekeeping gene expression levels between the diagnostic groups. Comparison of the expression level of the other genes confirmed ability to separate AML and ALL, whilst the direction of expression change (increased or decreased) for each gene between AML and ALL was the same as found by Golub et al. In particular, c-myb showed largest significant increase in ALL vs AML in the total BM fraction, whilst cystain c was increased in AML in the CD34−ve fraction. hSNF2b was significantly increased in the ALL total B.M fraction and Hox-A9 was significantly increased in the AML CD34+ve B.M fraction. Furthermore expression level of LYN and CD33 was significantly increased in AML compared to remission AML, indicating ability of the method to determine activity status of disease. In addition, several of the genes provided better separation between AML and ALL when measured in the CD34+ve and −ve fractions indicating more prominent expression in cells of different maturity and that prior fractionation is diagnostically more informative. The results demonstrate ability of the method to validate gene expression signatures by an independent method, which is simple, sensitive and robust, allowing translation to routine clinical use. Whilst the present study used AML and ALL, in principle the method could be extended to any other tumor type for which gene signatures exist.

scholarly journals Evaluation of GNMT Gene Expression in Prostate Cancer Tissues using Real-Time PCR

2021 ◽  
Author(s):  
Niloofar Dehghani ◽  
Masoud Salehipour ◽  
Babak Javanmard
Keyword(s):  
Gene Expression ◽  
Prostate Cancer ◽  
Benign Prostatic Hyperplasia ◽  
Real Time ◽  
Real Time Pcr ◽  
Prostatic Hyperplasia ◽  
Expression Level ◽  
P Value ◽  
Cancer Tissue ◽  
Tissue Samples

Introduction: Prostate cancer is the second most common cancer and the leading cause of cancer-related deaths worldwide. In the present study, the expression level of glycine N-methyl transferase gene (GNMT) was investigated in prostate cancer tissue. The GNMT enzyme is encoded by the GNMT gene. Increased GNMT gene expression increases the conversion of glycine to sarcosine and results in the elevated levels of sarcosine in blood and urine. Methods: The expression level of GNMT gene in tissue samples of patients with prostate cancer was compared with those with benign prostatic hyperplasia using Real-Time PCR technique. Results: The GNMT gene expression level increased significantly in prostate cancer patients compared with those with benign prostatic hyperplasia (p-value <0.001). In addition, the expression level of GNMT gene was stage-dependent and  significant increases were observed in all stages of prostate cancer compared with those with benign prostatic hyperplasia (p-value <0.001). Conclusion: The concentration of sarcosine is controlled by GNMT and it seems that increasing the expression level of GNMT gene increases the level of sarcosine concentration. Thus, it appears that increased levels of GNMT expression occur in the early stages of prostate cancer. Therefore, periodic measurement of GNMT expression levels can detect prostate cancer before it forms a cancer cell and invades other tissues.

scholarly journals A new set of endogenous control genes for use in quantitative real-time PCR experiments show that formin Ldia2dex transcripts are enriched in the early embryo of the pond snail Lymnaea stagnalis (Panpulmonata)

2019 ◽  
Author(s):  
Harriet F Johnson ◽  
Angus Davison
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Lymnaea Stagnalis ◽  
Model Organism ◽  
Early Embryo ◽  
Pond Snail ◽  
Endogenous Control ◽  
Quantitative Real Time Pcr ◽  
Nonsense Mediated Decay ◽  
Left Right Asymmetry

ABSTRACT Although the pond snail Lymnaea stagnalis is an emerging model organism for molecular studies in a wide variety of fields, there are a limited number of verified endogenous control genes for use in quantitative real-time PCR. As part of a larger study on snail chirality, or left–right asymmetry, we assayed gene expression in pond snail embryos. We evaluated six candidate control genes, by comparing their expression in three tissues (ovotestis, foot and embryo) and used three software programmes (geNorm, Normfinder and Bestkeeper) to do so. The specific utility of these control genes was then tested by investigating the relative expression of six experimental transcripts, including formin Ldia2, a gene that has been associated with chiral variation in L. stagnalis. All six control genes were found to be suitable for use in the three tissues tested. Of the six experimental genes, it was found that all were relatively depleted in the early embryo compared with other tissues, except the formin Ldia2 gene. Instead, transcripts of the wild-type Ldia2dex were enriched in the embryo, whereas a nonfunctional frameshifted version, Ldia2sin, was severely depleted. These differences in Ldia2sin expression were less evident in the ovotestis and were not evident in the foot tissue, possibly because nonsense-mediated decay is obscured in actively transcribing tissues. Our work provides a set of control genes that may be useful to the wider community and illustrates how these genes may be used to assay differences in expression in a variety of tissues.

scholarly journals Expression level of DREB gene of local corn cultivars from Kisar Island-Maluku, Indonesia, using quantitative real time polymerase chain reaction

2018 ◽  
Vol 9 (1) ◽  
Author(s):  
Hermalina Sinay ◽  
Estri Laras Arumingtyas
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Flag Leaf ◽  
Block Design ◽  
Rna Extraction ◽  
Expression Level ◽  
Total Rna ◽  
Deep Yellow ◽  
Dreb Gene

The research objective was to determine the expression level of dehydration responsive element binding (DREB) gene of local corn cultivars from Kisar Island Maluku, Indonesia. The study was designed as randomized block design with single factor consist of six local corn cultivars obtained from farmers in Kisar Island and one reference varieties which has been released as a drought-tolerant varieties and obtained from Cereal Crops Research Institute Maros South Sulawesi. Isolation of total RNA from the second leaf after the flag leaf at the 65 days after planting were carried out according to the protocols of the R and ABlueTM Total RNA Extraction Kit, and was used as a template for cDNA synthesis. Amplification of cDNA from total RNA was carried out according to the protocol of One-Step Reverse Transcriptase PCR Premix Kit. Real Time-PCR was performed usingcDNA from reverse transcription following the procedures of Real MODTM Green Real-Time PCR Master Mix Kit. The real time-PCR data were analyzed using relative quantification method based on the critical point/cycle threshold. The highest DREB gene expression was showed by Deep Yellow local corn cultivar, and the lowest one was showed by Rubby Brown Cob cultivar. The DREB gene expression level of deep yellow local corn cultivar was even higher than Srikandi variety as a reference variety.

scholarly journals A set of endogenous control genes for use in quantitative real-time PCR experiments reveal that the wild-type formin Ldia2 is enriched in the early pond snail embryo

2019 ◽  
Author(s):  
Harriet F. Johnson ◽  
Angus Davison
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Lymnaea Stagnalis ◽  
Model Organism ◽  
Early Embryo ◽  
Pond Snail ◽  
Endogenous Control ◽  
Wild Type ◽  
Quantitative Real Time Pcr ◽  
Nonsense Mediated Decay

ABSTRACTAlthough the pond snail Lymnaea stagnalis is an emerging model organism for molecular studies in a wide variety of fields including development, biomineralisation and neurophysiology, there are a limited number of verified endogenous control genes for use in quantitative real-time PCR (qRT-PCR). As part of larger study on snail chirality or left-right asymmetry, we wished to assay relative gene expression in pond snail embryos, so we evaluated six new candidate control genes, by comparing their expression in three tissues (ovotestis, foot, and embryo) and across three programs (geNorm, Normfinder and Bestkeeper). The specific utility of these control genes was then tested by investigating the relative expression of six experimental transcripts, including the formin Ldia2, a gene that has been associated with chirality in L. stagnalis. All six control genes were found to be suitable for use. Of the six experimental genes that were tested, it was found that all were relatively depleted in the early embryo compared with other tissues, except the formin gene Ldia2. Instead, transcripts of the wild type Ldia2dex were enriched in the embryo, whereas a non-functional frameshifted version Ldia2sin was severely depleted. These differences in Ldia2sin expression were less evident in the ovotestis and not evident in the foot tissue, suggesting that nonsense-mediated decay may be obscured in actively transcribing tissues. This work therefore provides a set of control genes that may be useful to the wider community, and shows how they may be used to assay differences in expression in the early embryo.

Respon Morfologis dan Ekspresi Gen Aquaporin pada Padi IR 64 yang Mengalami Cekaman Kekeringan pada Fase Reproduktif (Morphological Responses and Aquaporin Gene Expression in Rice IR64 under Drought Stress at the Reproductive Stage)

2018 ◽  
Vol 7 (2) ◽  
Author(s):  
Made Pharmawati ◽  
Ni Nyoman Wirasiti ◽  
Luh Putu Wrasiati
Keyword(s):  
Gene Expression ◽  
Drought Stress ◽  
Real Time ◽  
Real Time Pcr ◽  
Control Plant ◽  
Reproductive Stage ◽  
Limiting Factors ◽  
Rice Grain ◽  
Aquaporin Gene ◽  
Peg Treatment

Abstrak Cekaman kekeringan merupakan faktor pembatas penting bagi pertumbuhan dan produktivitas tanaman termasuk padi.      Penelitian ini bertujuan menganalisis respon padi IR64 terhadap cekaman kekeringan dengan pemberian polietilen glikol (PEG) pada fase reproduktif.  Penelitian juga bertujuan menganalisis ekspresi gen aquaporin akibat cekaman kekeringan.  Bibit padi ditanam dalam pot dan perlakuan PEG dengan konsentrasi 108g/L (-0.25MPa) dan 178g/L (-0.52 MPa) diberikan saat munculnya panikula. Perlakuan diberikan selama 2 minggu, kemudian tanaman disiram kembali.  Ekspresi gen diamati pada akhir perlakuan dengan semi kuantitatif real time PCR.  Ekstraksi RNA menggunakan RNeasy plant mini kit, sedangkan sintesis cDNA menggunakan Transcriptor First Strand cDNA Kit.  Hasil penelitian menunjukkan bahwa jumlah malai dan berat total malai berkurang akibat cekaman kekeringan.  Persentase gabah kosong mencapai 84,6% pada perlakuan PEG-0,52 MPa, sedangkan pada perlakuan PEG -0,25 MPa persentase gabah kosong sebesar 67,8%.  Pada kontrol persentase gabah kosong adalah 10,3%.  Ekspresi gen OsPIP2;7 sedikit menurun pada perlakuan PEG -0,52 MPa.Kata kunci: ekspresi gen, IR64, kekeringan, padi, PEG  Abstract Drought stress is one of the limiting factors of plant growth and productivity including rice.  The aim of this study was to analyze responses of IR64 rice to polyethylene glycol (PEG)-induced-drought stress at the reproductive stage.  This study also aimed to analyze the expression of aquaporin under drought stress.  Rice seedlings were grown in pot system and PEG treatment at concentration of -0.25MPa (108g/L) and -0.52 MPa (178g/L) were given when the panicles arose.  Treatments were conducted for 2 weeks, after that the plants were rewatered.  Gene expression was evaluated at the end of PEG treatment using semi quantitative real time PCR. RNA was extracted using RNeasy plant mini kit, while cDNA synthesis was done using Transcriptor First Strand cDNA Kit.  The results showed that the number and weight of rice ear were less in plant treated with PEG than in control.  The percentage of empty rice grain reached 84.6% at PEG -0.52 MPa, while at PEG -0.25 MPa the percentage of empty grain was 67.8%.  In control plant, the percentage of empty grain was 10.3%.  Drought stress did not alter the expression of OsPIP2;7.  Keywords: drought, gene expression, IR64, PEG, rice

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