scholarly journals Regulation of acetylcholine receptor alpha subunit variants in human myasthenia gravis. Quantification of steady-state levels of messenger RNA in muscle biopsy using the polymerase chain reaction.

1994 ◽  
Vol 94 (1) ◽  
pp. 16-24 ◽  
Author(s):  
T Guyon ◽  
P Levasseur ◽  
F Truffault ◽  
C Cottin ◽  
C Gaud ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Steady State ◽  
Myasthenia Gravis ◽  
Acetylcholine Receptor ◽  
Muscle Biopsy ◽  
Messenger Rna ◽  
Alpha Subunit ◽  
Chain Reaction ◽  
Steady State Levels ◽  
Polymerase Chain

Ectopic transcription of the parathyroid hormone gene in lymphocytes, lymphoblastoid cells and tumour tissue

1992 ◽  
Vol 135 (2) ◽  
pp. 249-256 ◽  
Author(s):  
O. Handt ◽  
A. Reis ◽  
J. Schmidtke
Keyword(s):  
Vitamin D ◽  
Polymerase Chain Reaction ◽  
Steady State ◽  
Parathyroid Hormone ◽  
Phorbol Esters ◽  
Rat Tissues ◽  
Chain Reaction ◽  
Lymphoblastoid Cells ◽  
Steady State Levels ◽  
Polymerase Chain

ABSTRACT Using reverse transcription polymerase chain reaction, steady-state levels of parathyroid hormone (PTH) mRNA were investigated in a number of human, bovine and rat tissues. Transcripts were consistently detected in parathyroid glands as well as in lymphocytes, lymphoblastoid cells and several tumours. Levels of transcription were not measurably increased in lymphoblastoid cells and tumour tissues compared with unstimulated peripheral lymphocytes. The level of 'ectopic' transcription of the PTH gene in lymphoblastoid cells appeared to be resistant to the administration of both vitamin D and phorbol esters. Journal of Endocrinology (1992) 135, 249–256

The effect of castration on steady state levels of luteinizing hormone-releasing hormone (LHRH) mRNA and proLHRH processing: time course study utilizing semi-quantitative reverse transcription/polymerase chain reaction

1996 ◽  
Vol 148 (3) ◽  
pp. 509-515 ◽  
Author(s):  
N V Emanuele ◽  
J Jurgens ◽  
N La Paglia ◽  
D W Williams ◽  
M R Kelley
Keyword(s):  
Polymerase Chain Reaction ◽  
Steady State ◽  
Luteinizing Hormone ◽  
Reverse Transcription ◽  
Male Rats ◽  
Chain Reaction ◽  
Luteinizing Hormone Releasing Hormone ◽  
Time Points ◽  
Steady State Levels ◽  
Polymerase Chain

Abstract Many studies have consistently shown that castration induces a prompt increase in serum levels and pituitary content of the gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), as well as a concomitant rise in steady state levels of the messenger RNAs directing their synthesis. The reports of effects of castration on the overall physiology of hypothalamic luteinizing hormone-releasing hormone (LHRH) — steady state levels of LHRH mRNA, post-translational processing and secretion — have, however, not been consistent. The goal of the studies reported here was to provide the first analysis of the effect of castration, at multiple post-operative time points, on steady state levels of LHRH mRNA and on the levels of hypothalamic proLHRH. All these data are correlated with hypothalamic levels of the mature LHRH decapeptide and with serum and pituitary levels of immunoreactive LH and FSH. Adult male rats were either castrated or sham-castrated (controls) and then sacrificed at 1, 3, 5, 7, 14, 21 or 28 days postoperatively. As expected, there was a prompt and sustained rise in serum immunoreactive LH and FSH in castrates compared with sham-operated animals. Intrapituitary LH levels rose above levels in the sham-operated animals by 14 days post castration. Intra-pituitary FSH showed a biphasic response, first falling significantly below control levels, then rising above control levels at 21 days. Steady state levels of LHRH mRNA in castrates, measured by reverse transcription/polymerase chain reaction, were increased about 2-fold above control levels by 1 day postoperatively, but were virtually identical to control levels at each of the other time points despite marked changes in the gonadotropins. ProLHRH content in castrates was 1·8-times that seen in controls at 1 day post castration (P<0·05), concomitant with the rise in steady state levels of LHRH mRNA at that time point. However, proLHRH content in castrates was no different from that seen in controls at each of the later time points examined. LHRH content was unchanged through 7 days after castration, but then fell significantly to 57% of control levels in hypothalami from animals gonadectomized 14 to 21 days previously (P<0·001 vs control), and to 54% of sham-operated levels at 28 days postoperatively (P<0·001). We conclude that: (1) changes in steady state levels of LHRH mRNA after castration are small and transient and (2) increased proLHRH coupled with unchanged LHRH levels at 1 day post castration, and castrate animal pro-LHRH at control levels coupled with falling LHRH at later post-castration time points indicate that the effect of gonadectomy on post-translational processing of pro-LHRH to LHRH is, likewise, small and transient. In aggregate our data suggest that most of the increase in serum LH and FSH seen in male rats after castration is not mediated at the hypothalamic level. Journal of Endocrinology (1996) 148, 509–515

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

scholarly journals Expression of DROSHA and DICER genes in peripheral blood leukocytes in lung sarcoidosis

2018 ◽  
Vol 90 (3) ◽  
pp. 21-24
Author(s):  
I E Malysheva ◽  
O V Balan ◽  
E L Tikhonovich ◽  
T O Volkova
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Peripheral Blood ◽  
Messenger Rna ◽  
Peripheral Blood Leukocytes ◽  
Chain Reaction ◽  
Blood Leukocytes ◽  
Healthy Donors ◽  
Polymerase Chain

Aim. To study the expression level of the genes DROSHA and DICER in peripheral blood leukocytes (PBL) of patients with sarcoidosis of the lungs Materials and methods. The study included 32 patients diagnosed with persistent lung sarcoidosis (mean age 41.56±1.27 years) and 36 healthy donors (control; mean age 42.79±1.95 years). The level of expression of messenger RNA (mRNA) of the genes DROSHA and DICER were determined in PBL of healthy donors and patients with sarcoidosis of the lung by polymerase chain reaction in real time. Results. As a result of the conducted researches it is established that the level of drosha gene expression in PBL patients with sarcoidosis of lungs is significantly reduced in comparison with the control (p

scholarly journals A beginner’s guide to RT-PCR, qPCR and RT-qPCR

2020 ◽  
Vol 42 (3) ◽  
pp. 48-53 ◽  
Author(s):  
Grace Adams
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
Life Science ◽  
Science Research ◽  
Quantitative Real Time Pcr ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Mrna Quantitation ◽  
Polymerase Chain

The development of the polymerase chain reaction (PCR), for which Kary Mullis received the 1992 Novel Prize in Chemistry, revolutionized molecular biology. At around the time that prize was awarded, research was being carried out by Russel Higuchi which led to the discovery that PCR can be monitored using fluorescent probes, facilitating quantitative real-time PCR (qPCR). In addition, the earlier discovery of reverse transcriptase (in 1970) laid the groundwork for the development of RT-PCR (used in molecular cloning). The latter can be coupled to qPCR, termed RT-qPCR, allowing analysis of gene expression through messenger RNA (mRNA) quantitation. These techniques and their applications have transformed life science research and clinical diagnosis.

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