214 research outputs found
Modularity map of the network of human cell differentiation
Cell differentiation in multicellular organisms is a complex process whose
mechanism can be understood by a reductionist approach, in which the individual
processes that control the generation of different cell types are identified.
Alternatively, a large scale approach in search of different organizational
features of the growth stages promises to reveal its modular global structure
with the goal of discovering previously unknown relations between cell types.
Here we sort and analyze a large set of scattered data to construct the network
of human cell differentiation (NHCD) based on cell types (nodes) and
differentiation steps (links) from the fertilized egg to a crying baby. We
discover a dynamical law of critical branching, which reveals a fractal
regularity in the modular organization of the network, and allows us to observe
the network at different scales. The emerging picture clearly identifies
clusters of cell types following a hierarchical organization, ranging from
sub-modules to super-modules of specialized tissues and organs on varying
scales. This discovery will allow one to treat the development of a particular
cell function in the context of the complex network of human development as a
whole. Our results point to an integrated large-scale view of the network of
cell types systematically revealing ties between previously unrelated domains
in organ functions.Comment: 32 pages, 7 figure
Two-photon resonant excitation of interatomic coulombic decay in neon dimers
The recent availability of intense and ultrashort extreme ultraviolet sources opens up the possibility of investigating ultrafast electronic relaxation processes in matter in an unprecedented regime. In this work we report on the observation of two-photon excitation of interatomic Coulombic decay (ICD) in neon dimers using the tunable intense pulses delivered by the free electron laser FERMI. The unique characteristics of FERMI (narrow bandwidth, spectral stability, and tunability) allow one to resonantly excite specific ionization pathways and to observe a clear signature of the ICD mechanism in the ratio of the ion yield created by Coulomb explosion. The present experimental results are explained by ab initio electronic structure and nuclear dynamics calculations
On a Generalization of Zaslavsky's Theorem for Hyperplane Arrangements
We define arrangements of codimension-1 submanifolds in a smooth manifold
which generalize arrangements of hyperplanes. When these submanifolds are
removed the manifold breaks up into regions, each of which is homeomorphic to
an open disc. The aim of this paper is to derive formulas that count the number
of regions formed by such an arrangement. We achieve this aim by generalizing
Zaslavsky's theorem to this setting. We show that this number is determined by
the combinatorics of the intersections of these submanifolds.Comment: version 3: The title had a typo in v2 which is now fixed. Will appear
in Annals of Combinatorics. Version. 2: 19 pages, major revision in terms of
style and language, some results improved, contact information updated, final
versio
Cisplatin and Oxaliplatin Toxicity: Importance of Cochlear Kinetics as a Determinant for Ototoxicity
Background
Cisplatin is a commonly used platinum anti-cancer drug. Regrettably cisplatin
has dose-limiting ototoxic side effects, e.g. the drug can induce an irreversible
hearing loss. The ototoxic mechanisms of cisplatin have not been
elucidated in the human ear and no clinically useful oto-protectors are yet
available. Cisplatin is a necessary part of many treatment regimes. Its beneficial
therapeutic effects might be reduced if cisplatin was excluded from the
treatment in order to protect the hearing function. In this work the ototoxic
effects of cisplatin are studied with the aim to better understand the mechanisms
behind the irreversible hearing loss induced by this drug. Oxaliplatin is
a second generation platinum-derivative anti-cancer drug, free from ototoxic
side effects in clinical practice. The effects of oxaliplatin on the inner ear have
been studied in this work and the results are compared with cisplatin treatment.
The two drugs differ regarding both anti-cancer effects and side effects,
which could be attributed to differences in pharmacokinetic factors, cellular
uptake and apoptotic mechanisms. The thioredoxin redox system with the
enzyme thioredoxin reductase (TrxR) was studied in cochleae due to a suggested
DNA-independent apoptotic mechanism of the hair cells. The cochlear
pharmacokinetics of cisplatin was assessed and the transport protein organic
cation transporter 2 (OCT2) was studied in relation to the ototoxic effect of
cisplatin.
Material and methods
Cultured human colon carcinoma cells and cell cultures of rat organ of Corti
were used for apoptosis studies in vitro following exposure to cisplatin and
oxaliplatin. Cisplatin and oxaliplatin were administered i.v. to guinea pigs,
followed by in vivo sampling of blood, cerebrospinal fluid (CSF) and scala
tympani (ST) perilymph. Liquid chromatography with post-column derivatization
was used to determine the concentration of parent drug in the samples.
Electrophysiological hearing thresholds and the loss of hair cells were assessed
to evaluate their ototoxic effects. Phenformin, a potential blocker of
OCT2 was administered and the ototoxic side effect of cisplatin was evaluated.
For immunohistochemical studies, cochlea from rat, guinea pig and pig
were used, where TrxR and OCT2 were evaluated in the cochlea. TrxR-assays
were used to measure the TrxR activity in cochlear tissue, both in vivo and in
vitro.
Results
The results from the in vitro studies showed that addition of either cisplatin
or oxaliplatin to the culture medium in organ of Corti cell cultures caused a
similar amount of outer hair cell loss and inhibition of TrxR activity. Cisplatin
exposure to cultured human colon carcinoma cells also reduced the activity
of TrxR. The results from the in vivo studies showed that a considerable concentration
of cisplatin was present in ST perilymph as compared with weak
concentrations of oxaliplatin after high dose oxaliplatin i.v. Ten minutes after
cisplatin administration, its concentration in ST perilymph was 4-fold higher
in the basal turn of the cochlea as compared to the apex. Cisplatin could be
analysed in ST perilymph for up to 120 min. Phenformin i.v. did not reduce
the ototoxic side-effect of cisplatin. Positive immunoreactivity to TrxR was
evident in both hair cells and spiral ganglion cells. Futhermore, OCT2 was
expressed in the supporting cells of organ of Corti and in the spiral ganglion
cells.
Conclusion
The transport of cisplatin to the vulnerable cells of hearing seems to be of major
importance for the ototoxic effects. An early high concentration of cisplatin
in the base of the cochlea and delayed elimination of cisplatin from ST perilymph
may be related to the cisplatin-induced loss of outer hair cells in the
basal turn of the cochlea. Cisplatin and oxaliplatin both cause similar ototoxic
effects when the organ of Corti is directly exposed in vitro. The thioredoxin
redox system with the TrxR enzyme may well play a critical role in cisplatininduced
ototoxicity. The presence of OCT2 in the supporting cells indicates
that this transport protein is primarily not involved in the uptake of cisplatin
from the systemic circulation but rather from the deeper compartments of
the cochlea. The knowledge elicited in this work will hopefully suggest objectives
for further studies in order to develop oto-protective treatments to
preserve the hearing of cisplatin treated patients
Apoptosis of the fibrocytes type 1 in the spiral ligament and blood labyrinth barrier disturbance cause hearing impairment in murine cerebral malaria
<p>Abstract</p> <p>Background</p> <p>Experimental murine malaria has been shown to result in significant hearing impairment. Microscopic evaluation of the temporal bones of these animals has revealed regular morphology of the cochlea duct. Furthermore, the known vascular pathologic changes being associated with malaria could not be found. Immunohistochemistry for ICAM1 showed a strong marking in the <it>stria vascularis</it>, indicating a disturbance of the endocochlear potential. The aim of this study was to evaluate the role of apoptosis and the disturbance of the blood labyrinth barrier in the murine malaria associated hearing impairment.</p> <p>Methods</p> <p>The temporal bones of seven mice with cerebral malaria-four with hearing impairment, three without hearing impairment-were evaluated with immunohistochemistry for cleaved caspase 3 to detect apoptosis and connexin 26, a gap junction protein being a cornerstone in the endocochlear potassium recirculation. Furthermore five animals with cerebral malaria were treated with Evans blue prior to sacrification to detect disturbances of the blood labyrinth barrier.</p> <p>Results</p> <p>Cleaved caspase 3 could clearly be detected by immunohistochemistry in the fibrocytes of the spiral ligament, more intensively in animals with hearing impairment, less intensively in those without. Apoptosis signal was equally distributed in the spiral ligament as was the connexin 26 gap junction protein. The Evans blue testing revealed a strong signal in the malaria animals and no signal in the healthy control animals.</p> <p>Conclusion</p> <p>Malfunction of the fibrocytes type 1 in the spiral ligament and disruption of the blood labyrinth barrier, resulting in a breakdown of the endocochlear potential, are major causes for hearing impairment in murine cerebral malaria.</p
Readthrough of nonsense mutations in Rett syndrome: evaluation of novel aminoglycosides and generation of a new mouse model
Thirty-five percent of patients with Rett syndrome carry nonsense mutations in the MECP2 gene. We have recently shown in transfected HeLa cells that readthrough of nonsense mutations in the MECP2 gene can be achieved by treatment with gentamicin and geneticin. This study was performed to test if readthrough can also be achieved in cells endogenously expressing mutant MeCP2 and to evaluate potentially more effective readthrough compounds. A mouse model was generated carrying the R168X mutation in the MECP2 gene. Transfected HeLa cells expressing mutated MeCP2 fusion proteins and mouse ear fibroblasts isolated from the new mouse model were treated with gentamicin and the novel aminoglycosides NB30, NB54, and NB84. The localization of the readthrough product was tested by immunofluorescence. Readthrough of the R168X mutation in mouse ear fibroblasts using gentamicin was detected but at lower level than in HeLa cells. As expected, the readthrough product, full-length Mecp2 protein, was located in the nucleus. NB54 and NB84 induced readthrough more effectively than gentamicin, while NB30 was less effective. Readthrough of nonsense mutations can be achieved not only in transfected HeLa cells but also in fibroblasts of the newly generated Mecp2R168X mouse model. NB54 and NB84 were more effective than gentamicin and are therefore promising candidates for readthrough therapy in Rett syndrome patients
New live screening of plant-nematode interactions in the rhizosphere
Abstract Free living nematodes (FLN) are microscopic worms found in all soils. While many FLN species are beneficial to crops, some species cause significant damage by feeding on roots and vectoring viruses. With the planned legislative removal of traditionally used chemical treatments, identification of new ways to manage FLN populations has become a high priority. For this, more powerful screening systems are required to rapidly assess threats to crops and identify treatments efficiently. Here, we have developed new live assays for testing nematode responses to treatment by combining transparent soil microcosms, a new light sheet imaging technique termed Biospeckle Selective Plane Illumination Microscopy (BSPIM) for fast nematode detection, and Confocal Laser Scanning Microscopy for high resolution imaging. We show that BSPIM increased signal to noise ratios by up to 60 fold and allowed the automatic detection of FLN in transparent soil samples of 1.5 mL. Growing plant root systems were rapidly scanned for nematode abundance and activity, and FLN feeding behaviour and responses to chemical compounds observed in soil-like conditions. This approach could be used for direct monitoring of FLN activity either to develop new compounds that target economically damaging herbivorous nematodes or ensuring that beneficial species are not negatively impacted
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