scholarly journals Expression of HERV-K proviruses in human leukocytes

Blood ◽  
1993 ◽  
Vol 81 (9) ◽  
pp. 2369-2374
Author(s):  
I Brodsky ◽  
B Foley ◽  
D Haines ◽  
J Johnston ◽  
K Cuddy ◽  
...  
Keyword(s):  
Human Blood ◽  
Blood Cells ◽  
Open Reading Frames ◽  
Chain Reaction ◽  
Blood Leukocytes ◽  
Normal Individuals ◽  
Class 1 ◽  
Polymerase Chain ◽  
Ribonuclease Protection ◽  
Reading Frames

HERV-K is a 50-copy, human endogenous, class 1 retroviral element that contains some polycistrons with gag, pol, and env open reading frames. Although expression of HERV-K proviruses has been shown in cultured human cell lines, expression of these elements has not been shown in human blood leukocytes. Using both reverse transcriptase-polymerase chain reaction and ribonuclease protection techniques, we show HERV-K pol gene expression in human blood leukocytes. Expression in blood leukocytes from 7 normal individuals was from a variety of different HERV-K proviruses, while restricted expression was observed in blood cells of 5 leukemia patients and 3 polycythemia vera patients. Evidence is presented suggesting that the restricted expression in leukemia blood cells is a result of gene regulation, not gene amplification.

scholarly journals Expression of HERV-K proviruses in human leukocytes

Blood ◽  
1993 ◽  
Vol 81 (9) ◽  
pp. 2369-2374 ◽  
Author(s):  
I Brodsky ◽  
B Foley ◽  
D Haines ◽  
J Johnston ◽  
K Cuddy ◽  
...  
Keyword(s):  
Human Blood ◽  
Blood Cells ◽  
Open Reading Frames ◽  
Chain Reaction ◽  
Blood Leukocytes ◽  
Normal Individuals ◽  
Class 1 ◽  
Polymerase Chain ◽  
Ribonuclease Protection ◽  
Reading Frames

Abstract HERV-K is a 50-copy, human endogenous, class 1 retroviral element that contains some polycistrons with gag, pol, and env open reading frames. Although expression of HERV-K proviruses has been shown in cultured human cell lines, expression of these elements has not been shown in human blood leukocytes. Using both reverse transcriptase-polymerase chain reaction and ribonuclease protection techniques, we show HERV-K pol gene expression in human blood leukocytes. Expression in blood leukocytes from 7 normal individuals was from a variety of different HERV-K proviruses, while restricted expression was observed in blood cells of 5 leukemia patients and 3 polycythemia vera patients. Evidence is presented suggesting that the restricted expression in leukemia blood cells is a result of gene regulation, not gene amplification.

scholarly journals Prevalence of GII.4 Sydney Norovirus Strains and Associated Factors of Acute Gastroenteritis in Children: 2019/2020 Season in Guangzhou, China

2021 ◽  
Author(s):  
Lei Duan ◽  
Xiaohan Yang ◽  
Jia Xie ◽  
Wenli Zhan ◽  
Changbin Zhang ◽  
...  
Keyword(s):  
Acute Gastroenteritis ◽  
Continuous Monitoring ◽  
Gene Sequence ◽  
Phylogenetic Analyses ◽  
Open Reading Frames ◽  
Severe Diarrhoea ◽  
Chain Reaction ◽  
Women And Children ◽  
Polymerase Chain ◽  
Reading Frames

AbstractNorovirus, the leading cause of non-bacterial acute gastroenteritis (AGE) worldwide, is constantly mutating. Continuous monitoring of the evolution of epidemic genotypes and emergence of novel genotypes is, therefore, necessary. This study determined the prevalence and clinical characteristics of norovirus strains in AGE in Guangzhou, China in 2019/2020 season. This study included children aged 2–60 months diagnosed with AGE in Guangzhou Women and Children Hospital, from August 2019 to January 2020. Norovirus was detected by real-time polymerase chain reaction and clinical data were obtained. Genotyping and phylogenetic analyses were performed with partial gene sequence fragments located within the open reading frames 1 and 2. During the study period, 168 children (61.3% males) were confirmed as norovirus infectious AGE. The main symptoms were diarrhoea and vomiting and 38 patients (22.6%) had seizures. Norovirus was mainly prevalent in October and November, and GII.4 Sydney[P31] was the major genotype circulating in Guangzhou. The phylogenetic tree showed that the Guangzhou strains had high homology with the strains circulating in 2017–2019 worldwide. GII.4 Sydney was the main prevalent norovirus genotype in Guangzhou from August 2019 to January 2020, which had more severe diarrhoea than those of other genotypes. These findings provide a valuable reference for the prevention, control, and treatment of norovirus in the future.

scholarly journals Analysis of pFQ12, a 22.4-kbFrankiaplasmid

2001 ◽  
Vol 47 (7) ◽  
pp. 608-617 ◽  
Author(s):  
Theodore R John ◽  
Jeffrey M Rice ◽  
Jerry D Johnson
Keyword(s):  
Amino Acids ◽  
Woody Plants ◽  
Codon Bias ◽  
Open Reading Frames ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Rich Region ◽  
Filter Hybridization ◽  
Polymerase Chain ◽  
Reading Frames

Frankia are gram-positive, filamentous bacteria capable of fixing atmospheric dinitrogen in symbiosis with a wide variety of woody plants and shrubs. Some isolates of Frankia harbor plasmids of 8.5 (pFQ11) and 22.4 kb (pFQ12) that have no known function but are transmitted through many generations in culture. We have sequenced the 22 437-bp pFQ12 plasmid that is present in isolates CpI1 and ArI3. This sequence, with 76% G+C, is almost totally unrelated to that of pFQ11 found in the same cells. However, four regions of identity, 40-90 bp each, are dispersed around the plasmids. The 22.4-kb plasmid has >50 open reading frames (ORFs) that encode putative proteins of more than 100 amino acids, with the largest being 2226 amino acids. Twenty of these ORFs are likely to encode proteins based on their codon bias as determined by two different algorithms. Transcripts from nine of these regions have been identified by reverse transcriptase-polymerase chain reaction (RT-PCR) or filter hybridization. The two Frankia plasmids each encode a protein similar to the korSA protein that regulates transmission of pSAM2 in Streptomyces. The origin of replication (ORI) region of pFQ12 was localized by intrastrand AT and GC equivalence switch. It includes a 40-bp, intergenic, A+T-rich region that has a strong identity in pFQ11.Key words: ORI analysis, RT-PCR, Glimmer, DNA sequence.

Detection of human cytomegalovirus DNA in peripheral blood leukocytes by the polymerase chain reaction

Transfusion ◽  
2003 ◽  
Vol 33 (9) ◽  
pp. 783-784 ◽  
Author(s):  
Ian.S. Bevan ◽  
Matthew R. Walker ◽  
Richard A. Daw ◽  
Andreas Bitsch ◽  
Gregor Bein ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Cytomegalovirus ◽  
Peripheral Blood ◽  
Peripheral Blood Leukocytes ◽  
Chain Reaction ◽  
Blood Leukocytes ◽  
Polymerase Chain

scholarly journals Peripheral Blood Leukocytes and Serum Nested Polymerase Chain Reaction Are Complementary Methods for Monitoring Active Cytomegalovirus Infection in Transplant Patients

2013 ◽  
Vol 24 (3) ◽  
pp. e69-e74 ◽  
Author(s):  
PD Andrade ◽  
MT Fioravanti ◽  
EBV Anjos ◽  
C De Oliveira ◽  
DM Albuquerque ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Positive Predictive Value ◽  
Predictive Value ◽  
Cytomegalovirus Infection ◽  
Clinical Symptoms ◽  
Peripheral Blood Leukocytes ◽  
Active Infection ◽  
Chain Reaction ◽  
Blood Leukocytes ◽  
Polymerase Chain

BACKGROUND: Human cytomegalovirus is an important cause of morbidity and mortality in immunocompromised patients. Qualitative polymerase chain reaction (PCR) has proven to be a sensitive and effective technique in defining active cytomegalovirus infection, in addition to having low cost and being a useful test for situations in which there is no need for quantification. Real-time PCR has the advantage of quantification; however, the high cost of this methodology makes it impractical for routine use.OBJECTIVE: To apply a nested PCR assay to serum (sPCR) and to evaluate its efficiency to diagnose active cytomegalovirus infection compared with PCR of peripheral blood leukocytes (L-PCR).METHODS: Samples of 37 patients were prospectively evaluated. An internal control was created and applied to sPCR to exclude false-negative results.RESULTS: In total, 21 patients (57%) developed active cytomegalovirus infection. After analyzing the two methods for the diagnosis of active infection, higher sensitivity and negative predictive value of the L-PCR versus sPCR (100% versus 62%), and higher specificity and positive predictive value of sPCR versus L-PCR (81% versus 50% and 72%, respectively) were observed. Discordant results were observed in 11 patients who were L-PCR-positive but sPCR-negative for active cytomegalovirus infection, five of whom developed clinical symptoms of cytomegalovirus. Clinical symptoms were observed in 14 patients, 12 of whom were diagnosed with active infection by nested L-PCR (P=0.007) and seven by nested sPCR (P=0.02). Higher specificity and a positive predictive value for sPCR were observed.CONCLUSION: Nested L-PCR and sPCR were considered to be complementary methods for the diagnosis and management of symptomatic cytomegalovirus infection.

scholarly journals Detection of the molecular abnormality in chronic myeloid leukemia by use of the polymerase chain reaction

Blood ◽  
1988 ◽  
Vol 72 (6) ◽  
pp. 2063-2065
Author(s):  
A Dobrovic ◽  
KJ Trainor ◽  
AA Morley
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Myeloid Leukemia ◽  
Blood Cells ◽  
Myeloid Leukemia ◽  
Lymphoblastic Leukemia ◽  
Leukemic Cell ◽  
Great Sensitivity ◽  
Chain Reaction ◽  
Molecular Abnormality ◽  
Polymerase Chain

The bcr-abl translocation characteristic of chronic myeloid leukemia (CML) was detected by the polymerase chain reaction (PCR) modified to use mRNA as the starting material. Amplification of a sequence spanning the bcr-abl junction was obtained by using peripheral blood cells from all of 20 patients with classic CML, one patient with acute lymphoblastic leukemia probably secondary to CML, and two cell lines derived from patients with CML. The presence of bcr exon 3 in the mRNA was determined from the size of the amplified sequence; it was present in 14 and absent in seven patients. One leukemic cell per 1,000 nonleukemic cells could be readily detected, thus indicating the great sensitivity of the method. This technique is of routine value in CML both for diagnosis and for following the course of treatment.

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 Congenital plasminogen deficiency caused by a Ser572 to Pro mutation

Blood ◽  
1993 ◽  
Vol 82 (2) ◽  
pp. 475-480 ◽  
Author(s):  
H Azuma ◽  
Y Uno ◽  
T Shigekiyo ◽  
S Saito
Keyword(s):  
Japanese Patient ◽  
Polymorphism Analysis ◽  
Chain Reaction ◽  
Japanese Family ◽  
Her Family ◽  
Restriction Fragment Polymorphism ◽  
Normal Individuals ◽  
Digestion Pattern ◽  
Polymerase Chain ◽  
Flanking Regions

Abstract We used a polymerase chain reaction (PCR) strategy and restriction fragment polymorphism analysis to evaluate all 19 exons of the plasminogen (PLG) gene in a Japanese patient with congenital PLG deficiency and her family members. She presented with cerebral infarction. Sequence analysis following amplification of each exon and its flanking regions showed a single T to C transition in exon 14, which changed a Ser572 codon (TCC) to Pro572 codon (CCC). Since this mutation generates a new Fok I site, the Fok I digestion pattern of the PCR-amplified exon 14 fragments from each family member was analyzed. In all cases, the patterns were consistent with the activities and antigen levels of plasma PLG in those members. Furthermore, all PCR- amplified exon 14 fragments from 15 normal individuals were not restricted with Fok I endonuclease. We conclude that a T to C transition in exon 14 identified in the propositus is responsible for PLG deficiency inherited in this Japanese family with thrombotic episodes.

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