2,197 research outputs found
Engineering Translation in Mammalian Cell Factories to Increase Protein Yield: The Unexpected Use of Long Non-Coding SINEUP RNAs
No full textAbstractMammalian cells are an indispensable tool for the production of recombinant proteins in contexts where function depends on post-translational modifications. Among them, Chinese Hamster Ovary (CHO) cells are the primary factories for the production of therapeutic proteins, including monoclonal antibodies (MAbs). To improve expression and stability, several methodologies have been adopted, including methods based on media formulation, selective pressure and cell- or vector engineering. This review presents current approaches aimed at improving mammalian cell factories that are based on the enhancement of translation. Among well-established techniques (codon optimization and improvement of mRNA secondary structure), we describe SINEUPs, a family of antisense long non-coding RNAs that are able to increase translation of partially overlapping protein-coding mRNAs. By exploiting their modular structure, SINEUP molecules can be designed to target virtually any mRNA of interest, and thus to increase the production of secreted proteins. Thus, synthetic SINEUPs represent a new versatile tool to improve the production of secreted proteins in biomanufacturing processes
Structural Properties of Polyglutamine Aggregates Investigated via Molecular Dynamics Simulations
No full textPolyglutamine (polyQ) beta-stranded aggregates constitute the hallmark of Huntington disease. The disease is fully penetrant when Q residues are more than 36-40 ("disease threshold"). Here, based on a molecular dynamics study on polyQ helical structures of different shapes and oligomeric states, we suggest that the stability of the aggregates increases with the number of monomers, while it is rather insensitive to the number of Qs in each monomer. However, the stability of the single monomer does depend on the number of side-chain intramolecular H-bonds, and therefore oil the number of Qs. If such number is lower than that of the disease threshold, the beta-stranded monomers are unstable and hence may aggregate with lower probability, consistently with experimental findings. Our results provide a possible interpretation of the apparent polyQ length dependent-toxicity, and they do not support the so-called "structural threshold hypothesis", which supposes a transition from random coil to a beta-sheet structure only above the disease threshold
"The love that made hell, paradise." Ouida re-writing the Paolo and Francesca theme in Held in Bondage
No full textThe bestselling Victorian author Ouida reveals in her novels, and, in particular, Held in Bondage, an extraordinary knowledge od Dante, by using characters and themes from the Commedia. The Paolo and Francesca theme actually constitutes part of the plot of the novel and is to be found in many of her other works, short stories and non-fiction writing
Entrapment into liposomes of fusicoccin binding sites
No full textFusicoccin (FC) binding sites solubilised from microsomal fractions of spinach leaves have been entrapped into soybean lecithin liposomes with an 80% yield. The investigation of the properties of these proteoliposomes has demonstrated that the rates of FC-binding and of exchange between radioactive and cold FC are intermediate between those observed with membrane-bound and with solubilised binding sites. It appears that the entrapped proteins are preferentially outside-oriented since they are inactivated by trypsin treatment; moreover, Triton X-100 permeabilization of the proteoliposomes demonstrated that one fifth of these sites are inside-oriented
HERStory Makers 2023: Francesca Fotheringham
No full textFrancesca Fotheringham is a postdoctoral research associate at the University of Edinburgh studying educational psychology with a focus on neurodiversity. She took part in HERStory Makers 2023.What is HERStory Makers?HERStory Makers is a social media competition for female-identifying early career researchers to share their research, their career journeys, and to inspire the next generation. Winners are selected by public vote. HERStory Makers is also part of EXPLORATHON, Scotland's contribution to European Researchers' Night.In 2022-23, EXPLORATHON Francescasupported by the Engineering & Physical Sciences Research Council [grant number EP/X020762/1].Author contributions to contentFrancesca conceived, planned, and recorded the video content. Kirsty Ross edited the video content to insert HERStory Maker credits, added subtitles, and reduce video length to below Twitter/X limit of 2 mins and 20 secs.</p
Medicina illuminata. La Biblioteca Lancisiana di Roma
No full textL'articolo presenta i codici miniati della Biblioteca Lancisiana di Roma. La prima parte, del coautore, è dedicata alla Biblioteca. La seconda parte, di F. Manzari, tratta dei manoscritti miniati, costituiti da due codici con le opere di Avicenna e dal Liber fraternitatis della Confraternita dell'Ospedale di Santo Spirito in Sassia a Roma.The article introduces the illuminated manuscripts of the Biblioteca Lancisiana in Rome. The first part of the article, by the co-author, is dedicated to the Library. The second part, by Francesca Manzari, illustrates the manuscipts; these are two manuscripts with the works of Avicenna and the Liber fraternitatis of the Confraternity of the Hospital of Santo Spirito in Sassia in Rome
Ser46 phosphorylation and prolyl-isomerase Pin1-mediated isomerization of p53 are key events in p53-dependent apoptosis induced by mutant huntingtin.
No full textHuntington disease (HD) is a neurodegenerative disorder caused by
a CAG repeat expansion in the gene coding for huntingtin protein.
Several mechanisms have been proposed by which mutant huntingtin
(mHtt) may trigger striatal neurodegeneration, including mitochondrial
dysfunction, oxidative stress, and apoptosis. Furthermore,
mHtt induces DNA damage and activates a stress response. In this
context, p53 plays a crucial role in mediatingmHtt toxic effects. Here
we have dissected the pathway of p53 activation by mHtt in human
neuronal cells and in HD mice, with the aim of highlighting critical
nodes that may be pharmacologically manipulated for therapeutic
intervention. We demonstrate that expression of mHtt causes increased
phosphorylation of p53 on Ser46, leading to its interaction
with phosphorylation-dependent prolyl isomerase Pin1 and consequent
dissociation from the apoptosis inhibitor iASPP, thereby
inducing the expression of apoptotic target genes. Inhibition of
Ser46 phosphorylation by targeting homeodomain-interacting protein
kinase 2 (HIPK2), PKCδ, or ataxia telangiectasia mutated kinase,
as well as inhibition of the prolyl isomerase Pin1, prevents mHttdependent
apoptosis of neuronal cells. These results provide a rationale
for the use of small-molecule inhibitors of stress-responsive
protein kinases and Pin1 as a potential therapeutic strategy for HD
treatment
Amyloid formation by mutant huntingtin: threshold, progressivity and recruitment of normal polyglutamine proteins
No full textHuntington's disease (HD) is caused by an expanded CAG trinucleotide repeat
encoding a tract of consecutive glutamines near the amino terminus of huntingtin,
a large protein of unknown function. It has been proposed that the expanded
polyglutamine stretch confers a new property on huntingtin and thereby causes
cell and region-specific neurodegeneration. Genotype-phenotype correlations
predict that this novel property appears above a threshold length (approximately
38 glutamines), becomes progressively more evident with increasing polyglutamine
length, is completely dominant over normal huntingtin and is not appreciably
worsened by a double genetic dose in HD homozygotes. Recently, an amino terminal
fragment of mutant huntingtin has been found to form self-initiated fibrillar
aggregates in vitro. We have tested the capacity for aggregation to assess
whether this property matches the criteria expected for a fundamental role in HD
pathogenesis. We find that that in vitro aggregation displays a threshold and
progressivity for polyglutamine length remarkably similar to the HD disease
process. Moreover, the mutant huntingtin amino terminus is capable of recruiting
into aggregates normal glutamine tract proteins, such as the amino terminal
segments of both normal huntingtin and of TATA-binding protein (TBP). Our
examination of in vivo aggregates from HD post-mortem brains indicates that they
contain an amino terminal segment of huntingtin of between 179 and 595 residues.
They also contain non-huntingtin protein, as evidenced by immunostaining for TBP.
Interestingly, like the in vitro aggregates, aggregates from HD brain display
Congo red staining with green birefringence characteristic of amyloid. Our data
support the view that the expanded polyglutamine segment confers on huntingtin a
new property that plays a determining role in HD pathogenesis and could be a
target for treatment. Moreover, the new property might have its toxic
consequences by interaction with one or more normal polyglutamine-containing
proteins essential for the survival of target neurons
Disequilibrium of multiple DNA markers on the human Y chromosome
No full textWe characterized four DNA polymorphisms on the Y chromosomes of 123 males from
five Caucasian populations. Three markers on the male specific portion of the
chromosome varied appreciably in frequency among the populations. When combined,
these markers define a limited number of haplotypes compared with the maximum
expected on the basis of random association. The associations found in the five
groups are qualitatively similar and are thus considered to be relatively stable
on an evolutionary time-scale and possibly to predate the divergence of Caucasian
populations. However, the haplotype frequencies varied markedly among
populations, even between weakly isolated areas such as northern vs. southern
Sardinia. This may indicate rapid progression towards fixation of alternative
types of Y chromosomes. We also report data suggesting that the same associations
no longer hold when examining a marker as close as 275 bp from the boundary of
the pseudoautosomal region on the Y chromosome
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