138 research outputs found
Spectroscopy and dissociative recombination of the lowest rotational states of H3+
The dissociative recombination of the lowest rotational states of H3+ has
been investigated at the storage ring TSR using a cryogenic 22-pole
radiofrequency ion trap as injector. The H3+ was cooled with buffer gas at ~15
K to the lowest rotational levels, (J,G)=(1,0) and (1,1), which belong to the
ortho and para proton-spin symmetry, respectively. The rate coefficients and
dissociation dynamics of H3+(J,G) populations produced with normal- and para-H2
were measured and compared to the rate and dynamics of a hot H3+ beam from a
Penning source. The production of cold H3+ rotational populations was
separately studied by rovibrational laser spectroscopy using chemical probing
with argon around 55 K. First results indicate a ~20% relative increase of the
para contribution when using para-H2 as parent gas. The H3+ rate coefficient
observed for the para-H2 source gas, however, is quite similar to the H3+ rate
for the normal-H2 source gas. The recombination dynamics confirm that for both
source gases, only small populations of rotationally excited levels are
present. The distribution of 3-body fragmentation geometries displays a broad
part of various triangular shapes with an enhancement of ~12% for events with
symmetric near-linear configurations. No large dependences on internal state or
collision energy are found.Comment: 10 pages, 9 figures, to be published in Journal of Physics:
Conference Proceeding
L1CAM Expression is Related to Non-Endometrioid Histology, and Prognostic for Poor Outcome in Endometrioid Endometrial Carcinoma
The majority of endometrial carcinomas are classified as Type I endometrioid endometrial carcinomas (EECs) and have a good prognosis. Type II non-endometrioid endometrial carcinomas (NEECs) have a significant worse outcome. Yet, 20Â % of the EECs are associated with an unexplained poor outcome. The aim of this study was to determine if L1CAM expression, a recently reported biomarker for aggressive tumor behavior in endometrial carcinoma, was associated with clinicopathological features of EECs. A total of 103 patients diagnosed as EEC at the Radboud University Medical Centre, based on the pathology report were selected. L1CAM status of these tumors was determined, and histologic slides were reviewed by two expert pathologists. L1CAM-positivity was observed in 17Â % (18/103). Review of the diagnostic slides revealed that 11 out of these 18 L1CAM-positive tumors (61Â %) contained a serous- or mixed carcinoma component that was not initially mentioned in the pathology report. L1CAM-expression was associated with advanced age, poor tumor grade, and lymphovascular space invasion. A worse five year progression free survival rate was observed for patients with L1CAM-positive tumors (55.6Â % for the L1CAM-positive group, compared to 83.3Â % for the L1CAM-negative group PÂ =Â 0.01). L1CAM expression carries prognostic value for histologically classified EEC and supports the identification of tumors with a NEEC component. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12253-016-0047-8) contains supplementary material, which is available to authorized users
Protocol for a systematic review of guidelines for rigour in the design, conduct and analysis of biomedical experiments involving laboratory animals
Objective: Within the last years, there has been growing awareness of the negative repercussions of unstandardized planning, conduct and reporting of preclinical and biomedical research. Several initiatives have set the aim of increasing validity and reliability in reporting of studies and publications, and publishers have formed similar groups. Additionally, several groups of experts across the biomedical spectrum have published experience and opinion-based guidelines and guidance on potential standardized reporting. While all these guidelines cover reporting of experiments, an important step prior to this should be rigours planning and conduction of studies. The aim of this systematic review is to identify and harmonize existing experimental design, conduct and analysis guidelines relating to internal validity and reproducibility of preclinical animal research. The review will also identify literature describing risks of bias pertaining to the design, conduct and analysis of preclinical biomedical research. Search strategy: PubMed, Embase and Web of Science will be searched systematically to identify guidelines published in English language in peer-reviewed journals before January 2018 (box 1). All articles or systematic reviews in English language that describe or review guidelines on the internal validity and reproducibility of animal studies will be included. Google search for guidelines published on the websites of major funders and professional organisations can be found in (Box 2). Screening and annotation: Unique references will be screened in two phases: screening for eligibility based on title and abstract, followed by screening for definitive inclusion based on full text. Screening will be performed in SyRF (http://syrf.org.uk). Each reference will be randomly presented to two independent reviewers. Disagreements between reviewers will be resolved by additional screening of the reference by a third, senior researcher. Data management and reporting: All data, including extracted text and guidelines, will be stored in the SyRF platform. Elements of the included guidelines will be identified using a standardized extraction form. Reporting will follow the PRISMA guidelines as far as applicable
Assignment of resonances in dissociative recombination of HD+ ions: high-resolution measurements compared with accurate computations
The collision-energy resolved rate coefficient for dissociative recombination
of HD+ ions in the vibrational ground state is measured using the photocathode
electron target at the heavy-ion storage ring TSR. Rydberg resonances
associated with ro-vibrational excitation of the HD+ core are scanned as a
function of the electron collision energy with an instrumental broadening below
1 meV in the low-energy limit. The measurement is compared to calculations
using multichannel quantum defect theory, accounting for rotational structure
and interactions and considering the six lowest rotational energy levels as
initial ionic states. Using thermal equilibrium level populations at 300 K to
approximate the experimental conditions, close correspondence between
calculated and measured structures is found up to the first vibrational
excitation threshold of the cations near 0.24 eV. Detailed assignments,
including naturally broadened and overlapping Rydberg resonances, are performed
for all structures up to 0.024 eV. Resonances from purely rotational excitation
of the ion core are found to have similar strengths as those involving
vibrational excitation. A dominant low-energy resonance is assigned to
contributions from excited rotational states only. The results indicate strong
modifications in the energy dependence of the dissociative recombination rate
coefficient through the rotational excitation of the parent ions, and underline
the need for studies with rotationally cold species to obtain results
reflecting low-temperature ionized media.Comment: 15 pages, 10 figures. Paper to appear in Phys. Rev. A (version as
accepted
Resonant structure of low-energy H3+ dissociative recombination
New high-resolution dissociative recombination rate coefficients of
rotationally cool and hot H3+ in the vibrational ground state have been
measured with a 22-pole trap setup and a Penning ion source, respectively, at
the ion storage ring TSR. The experimental results are compared with
theoretical calculations to explore the dependence of the rate coefficient on
ion temperature and to study the contributions of different symmetries to probe
the rich predicted resonance spectrum. The break-up energy was investigated by
fragment imaging to derive internal temperatures of the stored parent ions
under differing experimental conditions. A systematic experimental assessment
of heating effects is performed which, together with a survey of other recent
storage-ring data, suggests that the present rotationally cool rate-coefficient
measurement was performed at 380^{+50}_{-130} K and that this is the lowest
rotational temperature so far realized in storage-ring rate-coefficient
measurements on H3+. This partially supports the theoretical suggestion that
higher temperatures than assumed in earlier experiments are the main cause for
the large gap between the experimental and theoretical rate coefficients. For
the rotationally hot rate-coefficient measurement a temperature of below 3250K
is derived. From these higher-temperature results it is found that increasing
the rotational ion temperature in the calculations cannot fully close the gap
between the theoretical and experimental rate coefficients.Comment: 12 pages, 7 figures (11 subfigures), 3 table
Pathologic and Phenotypic Alterations in a Mouse Expressing a Connexin47 Missense Mutation That Causes Pelizaeus-Merzbacher–Like Disease in Humans
Gap junction channels are intercellular conduits that allow diffusional exchange of ions, second messengers, and metabolites. Human oligodendrocytes express the gap junction protein connexin47 (Cx47), which is encoded by the GJC2 gene. The autosomal recessive mutation hCx47M283T causes Pelizaeus-Merzbacher–like disease 1 (PMLD1), a progressive leukodystrophy characterized by hypomyelination, retarded motor development, nystagmus, and spasticity. We introduced the human missense mutation into the orthologous position of the mouse Gjc2 gene and inserted the mCx47M282T coding sequence into the mouse genome via homologous recombination in embryonic stem cells. Three-week-old homozygous Cx47M282T mice displayed impaired rotarod performance but unchanged open-field behavior. 10-15-day-old homozygous Cx47M282T and Cx47 null mice revealed a more than 80% reduction in the number of cells participating in glial networks after biocytin injections into oligodendrocytes in sections of corpus callosum. Homozygous expression of mCx47M282T resulted in reduced MBP expression and astrogliosis in the cerebellum of ten-day-old mice which could also be detected in Cx47 null mice of the same age. Three-month-old homozygous Cx47M282T mice exhibited neither altered open-field behavior nor impaired rotarod performance anymore. Adult mCx47M282T expressing mice did not show substantial myelin alterations, but homozygous Cx47M282T mice, additionally deprived of connexin32, which is also expressed in oligodendrocytes, died within six weeks after birth and displayed severe myelin defects accompanied by astrogliosis and activated microglia. These results strongly suggest that PMLD1 is caused by the loss of Cx47 channel function that results in impaired panglial coupling in white matter tissue
Interaction and uptake of exosomes by ovarian cancer cells
<p>Abstract</p> <p>Background</p> <p>Exosomes consist of membrane vesicles that are secreted by several cell types, including tumors and have been found in biological fluids. Exosomes interact with other cells and may serve as vehicles for the transfer of protein and RNA among cells.</p> <p>Methods</p> <p>SKOV3 exosomes were labelled with carboxyfluoresceine diacetate succinimidyl-ester and collected by ultracentrifugation. Uptake of these vesicles, under different conditions, by the same cells from where they originated was monitored by immunofluorescence microscopy and flow cytometry analysis. Lectin analysis was performed to investigate the glycosylation properties of proteins from exosomes and cellular extracts.</p> <p>Results</p> <p>In this work, the ovarian carcinoma SKOV3 cell line has been shown to internalize exosomes from the same cells via several endocytic pathways that were strongly inhibited at 4°C, indicating their energy dependence. Partial colocalization with the endosome marker EEA1 and inhibition by chlorpromazine suggested the involvement of clathrin-dependent endocytosis. Furthermore, uptake inhibition in the presence of 5-ethyl-N-isopropyl amiloride, cytochalasin D and methyl-beta-cyclodextrin suggested the involvement of additional endocytic pathways. The uptake required proteins from the exosomes and from the cells since it was inhibited after proteinase K treatments. The exosomes were found to be enriched in specific mannose- and sialic acid-containing glycoproteins. Sialic acid removal caused a small but non-significant increase in uptake. Furthermore, the monosaccharides D-galactose, α-L-fucose, α-D-mannose, D-N-acetylglucosamine and the disaccharide β-lactose reduced exosomes uptake to a comparable extent as the control D-glucose.</p> <p>Conclusions</p> <p>In conclusion, exosomes are internalized by ovarian tumor cells via various endocytic pathways and proteins from exosomes and cells are required for uptake. On the other hand, exosomes are enriched in specific glycoproteins that may constitute exosome markers. This work contributes to the knowledge about the properties and dynamics of exosomes in cancer.</p
Full-Length L1CAM and Not Its Δ2Δ27 Splice Variant Promotes Metastasis through Induction of Gelatinase Expression
Tumour-specific splicing is known to contribute to cancer progression. In the case of the L1 cell adhesion molecule (L1CAM), which is expressed in many human tumours and often linked to bad prognosis, alternative splicing results in a full-length form (FL-L1CAM) and a splice variant lacking exons 2 and 27 (SV-L1CAM). It has not been elucidated so far whether SV-L1CAM, classically considered as tumour-associated, or whether FL-L1CAM is the metastasis-promoting isoform. Here, we show that both variants were expressed in human ovarian carcinoma and that exposure of tumour cells to pro-metastatic factors led to an exclusive increase of FL-L1CAM expression. Selective overexpression of one isoform in different tumour cells revealed that only FL-L1CAM promoted experimental lung and/or liver metastasis in mice. In addition, metastasis formation upon up-regulation of FL-L1CAM correlated with increased invasive potential and elevated Matrix metalloproteinase (MMP)-2 and -9 expression and activity in vitro as well as enhanced gelatinolytic activity in vivo. In conclusion, we identified FL-L1CAM as the metastasis-promoting isoform, thereby exemplifying that high expression of a so-called tumour-associated variant, here SV-L1CAM, is not per se equivalent to a decisive role of this isoform in tumour progression
Tumor-derived microRNAs induce myeloid suppressor cells and predict immunotherapy resistance in melanoma
The accrual of myeloid-derived suppressor cells (MDSCs) represents a major obstacle to effective immunotherapy in cancer patients, but the mechanisms underlying this process in the human setting remain elusive. Here, we describe a set of microRNAs (miR-146a, miR-155, miR-125b, miR-100, let-7e, miR-125a, miR-146b, miR-99b) that are associated with MDSCs and with resistance to treatment with immune checkpoint inhibitors in melanoma patients. The miRs were identified by transcriptional analyses as being responsible for the conversion of monocytes into MDSCs (CD14+HLA-DRneg cells) mediated by melanoma extracellular vesicles (EVs) and were shown to recreate MDSC features upon transfection. In melanoma patients, these miRs are increased in circulating CD14+ monocytes, plasma and tumor samples, where they correlate with the myeloid cell infiltrate. In plasma, their baseline level clusters with the clinical efficacy of CTLA-4 or PD-1 blockade. Hence, MDSC-related miRs represent an indicator of MDSC activity in cancer patients and a potential blood marker of a poor immunotherapy outcome
- …