4,207 research outputs found
Digital multiplexed mRNA analysis of functionally important genes in single human oocytes and correlation of changes in transcript levels with oocyte protein expression
Objective
To investigate functionally important transcripts in single human oocytes with the use of NanoString technology and determine whether observed differences are biologically meaningful.
Design
Analysis of human oocytes with the use of NanoString and immunoblotting.
Setting
University-affiliated reproductive medicine unit.
Patients
Women undergoing in vitro fertilization.
Intervention
Human oocytes were analyzed with the use of NanoString or immunoblotting.
Main Outcome Measures
The abundance of transcripts for ten functionally important genes—AURKA, AURKC, BUB1, BUB1B (encoding BubR1), CDK1, CHEK1, FYN, MOS, MAP2K1, and WEE2—and six functionally dispensable genes were analyzed with the use of NanoString. BubR1 protein levels in oocytes from younger and older women were compared with the use of immunoblotting.
Result(s)
All ten functional genes but none of the six dispensable genes were detectable with the use of NanoString in single oocytes. There was 3- to 5-fold variation in BUB1, BUB1B, and CDK1 transcript abundance among individual oocytes from a single patient. Transcripts for these three genes—all players within the spindle assembly checkpoint surveillance mechanism for preventing aneuploidy—were reduced in the same oocyte from an older patient. Mean BUB1B transcripts were reduced by 1.5-fold with aging and associated with marked reductions in BubR1 protein levels.
Conclusion(s)
The abundance of functionally important transcripts exhibit marked oocyte-to-oocyte heterogeneity to a degree that is sufficient to affect protein expression. Observed variations in transcript abundance are therefore likely to be biologically meaningful, especially if multiple genes within the same pathway are simultaneously affected
How to write an accounting history book review
The purpose of a book review is to communicate both the delight and the importance of a book. Books have been defined by the Supreme Court in terms of an author\u27s intellectual production and not in terms of the particular form which such production ultimately takes. Language : en
Quantum effects from a purely geometrical relativity theory
A purely geometrical relativity theory results from a construction that
produces from three-dimensional space a happy unification of Kaluza's
five-dimensional theory and Weyl's conformal theory. The theory can provide
geometrical explanations for the following observed phenomena, among others:
(a) lifetimes of elementary particles of lengths inversely proportional to
their rest masses; (b) the equality of charge magnitude among all charged
particles interacting at an event; (c) the propensity of electrons in atoms to
be seen in discretely spaced orbits; and (d) `quantum jumps' between those
orbits. This suggests the possibility that the theory can provide a
deterministic underpinning of quantum mechanics like that provided to
thermodynamics by the molecular theory of gases.Comment: 7 pages, LaTeX jpconf.cls (Institute of Physics Publishing), 6
Encapsulated PostScript figures (Fig. 6 is 1.8M uncompressed); Presented at
VI Mexican School on Gravitation and Mathematical Physics "Approaches to
Quantum Gravity
An orthologue of bacteroides fragilis NanH is the principal sialidase in tannerella forsythia
Sialidase activity is a putative virulence factor of the anaerobic periodontal pathogen Tannerella forsythia, but it is uncertain which genes encode this activity. Characterization of a putative sialidase, SiaHI, by others, indicated that this protein alone may not be responsible for all of the sialidase activity. We describe a second sialidase in T. forsythia (TF0035), an orthologue of Bacteroides fragilis NanH, and its expression in Escherichia coli. Sialidase activity of the expressed NanH was confirmed by using 2′-(4-methylumbelliferyl)-α-d-N-acetylneuraminic acid as a substrate. Biochemical characterization of the recombinant T. forsythia NanH indicated that it was active over a broad pH range, with optimum activity at pH 5.5. This enzyme has high affinity for 2′-(4-methylumbelliferyl)-α-d-N-acetylneuraminic acid (Km of 32.9 ± 10.3 μM) and rapidly releases 4-methylumbelliferone (Vmax of 170.8 ± 11.8 nmol of 4-methylumbelliferone min−1 mg of protein−1). E. coli lysates containing recombinant T. forsythia NanH cleave sialic acid from a range of substrates, with a preference for α2-3 glycosidic linkages. The genes adjacent to nanH encode proteins apparently involved in the metabolism of sialic acid, indicating that the NanH sialidase is likely to be involved in nutrient acquisition
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