Abstract:
:We describe a rapid and specific differential amplification system which can detect five of the most common cystic fibrosis mutations from a single cell. In the first round of the polymerase chain reaction (PCR), regions of exons 4, 10 and 11 of the cystic fibrosis transmembrane conductance regulator (CFTR) gene containing the mutations delta F508, G551D, R553X, G542X and 621+1G > T were co-amplified in a single multiplex PCR. To identify potential contamination, we included external amplification primers for the polymorphic human tyrosine hydroxylase (HUMTH01) locus as a fingerprint for the sample. In the second round of PCR, detection of any of the five mutations was achieved using the amplification refractory mutation system (ARMS) in two separate reactions, each containing nested amplification primers for either wild type or mutant sequence. A separate second round PCR for the fingerprinting was performed with nested HUMTH01 primers. Using this procedure we have successfully and accurately detected five cystic fibrosis mutations in 30 single cells with a failed amplification rate of 7% and a contamination rate of 4.6% and that PCR failure or possible contamination will also be easily detected. This procedure allows detection of the five most common point mutations and small deletions responsible for cystic fibrosis from a single cell in < 8 h which could be applicable to preimplantation diagnosis in human embryos.
journal_name
Mol Hum Reprodjournal_title
Molecular human reproductionauthors
Scobie G,Woodroffe B,Fishel S,Kalsheker Ndoi
10.1093/molehr/2.3.203subject
Has Abstractpub_date
1996-03-01 00:00:00pages
203-7issue
3eissn
1360-9947issn
1460-2407journal_volume
2pub_type
杂志文章abstract::Preterm premature rupture of the membranes (PPROM) has been considered to be closely associated with chorioamnionitis. However, the detailed mechanism is not well understood. Alpha 1 antitrypsin (AAT) was reported to decrease in concentration in amniotic fluid obtained from patients with PPROM. However, the origin of ...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gan071
更新日期:2009-01-01 00:00:00
abstract:STUDY QUESTION:Are there novel hyaladherins in human sperm? SUMMARY ANSWER:Zona pellucida-binding protein 2 (ZPBP2), containing a Link-like hyaluronic acid (HA)-binding domain, and several other proteins containing BX7B motifs, such as ADAM32 and Midkine, may be novel hyaladherins with HA-binding properties. WHAT IS ...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gax053
更新日期:2017-12-01 00:00:00
abstract::The etiology and pathogenesis of endometriosis are complex with both genetic and environmental factors contributing to disease risk. Genome-wide association studies (GWAS) have identified multiple signals in the estrogen receptor 1 (ESR1) region associated with endometriosis and other reproductive traits and diseases....
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gaaa082
更新日期:2021-01-22 00:00:00
abstract::Tenascin-C is an extracellular matrix glycoprotein whose monomers include eight consecutive fibronectin type III-like repeats, encoded by exons 10-16, and which are subject to alternative splicing. Transcripts containing these exons are expressed during tissue wounding and active tissue remodelling. Human fetal membra...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/5.11.1066
更新日期:1999-11-01 00:00:00
abstract::We have previously reported that endometrial expression of matrix metalloproteinase (MMP)-26 mRNA comes to a maximum in the early secretory phase. Since tissue inhibitor of metalloproteinase (TIMP)-4 is a potent inhibitor of MMP-26, the objective of this study was to identify the pattern of TIMP-4 mRNA expression in t...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gah092
更新日期:2004-09-01 00:00:00
abstract::Fetal membranes overlying the cervix in patients prior to and during labour, and within the rupture tear after spontaneous delivery at term, exhibit altered morphology. In this study we report that in comparison to mid-zone fetal membranes biopsies, these regions are characterized by increased expression of the matric...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/7.5.463
更新日期:2001-05-01 00:00:00
abstract::We have investigated the possible roles of oncostatin M (OSM), which is a member of the interleukin-6 family of cytokines, in endometrial and endometriotic stromal cell growth. Endometrial and endometriotic stromal cells were collected from the uterus or ovarian chocolate cysts. We observed the expression of mRNA tran...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/7.7.665
更新日期:2001-07-01 00:00:00
abstract:STUDY QUESTION:How does hCG signal in human endometrial stromal cells (ESCs) and what is its role in regulating ESC function? SUMMARY ANSWER:hCG signaling in ESCs activates the extracellular signal-regulated protein kinases 1 and 2 (Erk1/2) pathway through exchange protein activated by cyclic AMP (cAMP) (Epac) and tra...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gax015
更新日期:2017-06-01 00:00:00
abstract::After meiosis, during the final stages of spermatogenesis, the haploid male genome undergoes major structural changes, resulting in a shift from a nucleosome-based genome organization to the sperm-specific, highly compacted nucleoprotamine structure. Recent data support the idea that region-specific programming of the...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gar063
更新日期:2012-01-01 00:00:00
abstract::Within the context of an oocyte in vitro maturation (IVM) program for reproductive treatment, oocyte cumulus complexes (COCs) derived from follicles <6 mm in patients with PCOS were matured in vitro. Key transcripts related to meiotic maturation (FSHR, LHCGR, EGFR, PGR) and oocyte competence (AREG, ADAMTS, HAS2, PTGS2...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gas038
更新日期:2013-01-01 00:00:00
abstract::This study was undertaken to evaluate the effects of gamma-aminobutyric acid (GABA) and GABAergic agonists and antagonists on sperm kinematic parameters and hyperactivation, evaluated by a computer-assisted semen analysis (CASA) system, and intracellular cAMP content in 22 normozoospermic semen samples. Because of the...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/2.10.733
更新日期:1996-10-01 00:00:00
abstract::Oocyte vitrification has been introduced into clinical settings without extensive pre-clinical safety testing. In this study, we analysed major safety aspects of human oocyte vitrification in a high security closed system: (i) chromosomal meiotic segregation, (ii) embryonic developmental kinetics and (iii) DNA (hydrox...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gav013
更新日期:2015-06-01 00:00:00
abstract::Mitochondrial DNA (mtDNA) deletions are present in both human oocytes and embryos. It has been found that these tissues contain a mtDNA mutation which is present in high amounts in patients with Kearns-Sayre syndrome (KSS) and progressive external ophthalmoplegia. In the present study, the frequency of this KSS deleti...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/4.9.887
更新日期:1998-09-01 00:00:00
abstract::The endocrine disrupting chemical o, p'-dichlorodiphenyltrichloroethane (DDT) can affect reproductive organs, tissues and cells in several species. Treatment of human endometrial endothelial cells (HEECs) with 50 microM o,p'-DDT decreased their proliferation compared with the control. Microarray analyses revealed that...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gam091
更新日期:2008-02-01 00:00:00
abstract::Recent data demonstrated that CD4+CD25+ regulatory T (Treg) cells and an enzyme called indoleamine 2,3-dioxygenase (IDO) mediate maternal tolerance to the fetus. Interestingly, Treg cells express the CTLA-4 molecule on their surface, and B7 (CD80/86) ligation by CTLA-4 enhanced IDO activity of dendritic cells (DCs) an...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gah246
更新日期:2005-12-01 00:00:00
abstract::The requisite endpoint of mammalian oocyte maturation, whether in vivo or in vitro, is a metaphase II oocyte which is able to be fertilized and which can eventually support normal embryonic development. Oocytes which have been matured in vivo basically fulfill these criteria. On the other hand, a completely different ...
journal_title:Molecular human reproduction
pub_type: 杂志文章,评审
doi:10.1093/molehr/4.1.41
更新日期:1998-01-01 00:00:00
abstract::During early gestation, a considerable increase in different leukocyte subsets can be observed in the decidualized endometrium concomitantly to the invasion of cytotrophoblast cells (CTB). To date, it is still in question which factors induce this accumulation of immune cells and whether it is evoked by an in situ pro...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gat005
更新日期:2013-06-01 00:00:00
abstract::Successful intracytoplasmic sperm injection (ICSI) is dependent upon the competence of the oocyte to respond to the injection of a spermatozoon in the presence of calcium. This study determined if oocytes that failed to become fertilized (absence of any pronuclei) failed to undergo cytoplasmic activation, as ascertain...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/2.12.959
更新日期:1996-12-01 00:00:00
abstract::In the implantation, trophoblasts penetrate maternal decidua by secreting proteases. It has been reported that cathepsins are highly expressed in the mouse villi, and play an important role in normal embryonal growth and decidualization. In this study, we evaluated cathepsins and their endogenous inhibitors, cystatins...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gah172
更新日期:2005-05-01 00:00:00
abstract::Successful pregnancy in humans depends on deep invasion of the maternal decidua by extravillous trophoblast cells (EVTs), a process regulated by autocrine and paracrine signals in the decidual-trophoblast microenvironment. Here we examined whether trophoblast invasion is affected by decidual differentiation of endomet...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gar015
更新日期:2011-07-01 00:00:00
abstract::Stromal cell-derived factor-1 (SDF-1 or CXCL12) is the physiologic ligand for the chemokine receptor CXCR4. CXCR4-mediated signalling regulates cell migration and apoptosis in certain haematopoietic and neuronal cells. Using gene profiling, we determined that CXCR4 is the only chemokine receptor for which mRNA express...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gah118
更新日期:2004-12-01 00:00:00
abstract:STUDY QUESTION:Which different pathways and functions are altered in rhesus monkey oocytes that fail to mature after an ovulatory stimulus? SUMMARY ANSWER:Failed to mature (FTM) oocytes complete a large portion of the transition in transcriptome composition associated with normal maturation, but also manifest numerous...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gay032
更新日期:2018-10-01 00:00:00
abstract::In recent years, the impact of sperm DNA damage on fertility has become an important issue. The different technologies developed to check sperm DNA fragmentation lead to the same conclusion: DNA damage negatively impacts upon reproductive processes. Oocyte DNA repair capacity is one of the cues to understanding embryo...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gam038
更新日期:2007-08-01 00:00:00
abstract::Medium-chain acyl-CoA dehydrogenase (MCAD) deficiency is the most common defect in fatty acid oxidation. The disease is inherited in an autosomal recessive fashion (carrier frequency around 1 in 70) and probably affects as many as 1 in 10000 new-borns. Affected children usually present within the two first years of li...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/6.12.1165
更新日期:2000-12-01 00:00:00
abstract::The molecular mechanism of stress-associated reproductive dysfunction is complex and largely unknown. This study was designed to systematically analyze molecular effects of systemic in vivo blockade of α1-adrenergic receptors (α1-ADRs) on stress-induced disturbance of cAMP/cGMP signaling in testosterone-producing Leyd...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gat052
更新日期:2014-01-01 00:00:00
abstract::Oocytes are usually the largest cells in the body and as such offer unique opportunities for single-cell analysis. Unfortunately, these cells are also some of the rarest in the mammalian female, usually necessitating single-cell analysis. In cases of infertility in humans, determining the quality of the oocyte is ofte...
journal_title:Molecular human reproduction
pub_type: 杂志文章,评审
doi:10.1093/molehr/gav064
更新日期:2016-03-01 00:00:00
abstract::A major challenge in the comprehension of the endometrial transformations leading to the completion of each menstrual cycle in humans is in the identification of specific molecular pathways underlying these monthly turnovers. Towards this goal we compared, by the differential display technique, the relative expression...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gag084
更新日期:2003-11-01 00:00:00
abstract::Patients with rare overgrowth disorders, such as Beck-Wiedemann syndrome and Simpson-Golabi-Behmel syndrome, are predisposed to embryonal tumours, including Wilms' tumour of the kidney. Therefore, these disorders offer a link between hyperplastic growth and cancer. Genetic lesions at chromosome 11p15 have been associa...
journal_title:Molecular human reproduction
pub_type: 杂志文章,评审
doi:10.1093/molehr/3.2.157
更新日期:1997-02-01 00:00:00
abstract::Endometriosis is a gynaecological disease with a certain genetic background, but the locations of possible genomic aberrations are still poorly clarified. Intercellular adhesion molecule-1 (ICAM-1), which is a surface glycoprotein that promotes adhesion in immunological and inflammatory reactions, seems to play a role...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gag002
更新日期:2003-01-01 00:00:00
abstract::In the uterus, epithelial cell proliferation changes during the estrous cycle and pregnancy. Uncontrolled epithelial cell proliferation results in implantation failure and/or cancer development. Transforming growth factor-β (TGF-β) signaling is a fundamental regulator of diverse biological processes and is indispensab...
journal_title:Molecular human reproduction
pub_type: 杂志文章
doi:10.1093/molehr/gau033
更新日期:2014-08-01 00:00:00