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Hester J. Scheffer Jantien A. Vogel Willemien van den Bos Robert E. Neal II Krijn P. van Lienden Marc G. H. Besselink Martin J. C. van Gemert Cees W. M. van der Geld Martijn R. Meijerink John H. Klaessens Rudolf M. Verdaasdonk 《PloS one》2016,11(2)
Purpose
Irreversible electroporation (IRE) uses short duration, high-voltage electrical pulses to induce cell death via nanoscale defects resulting from altered transmembrane potential. The technique is gaining interest for ablations in unresectable pancreatic and hepatobiliary cancer. Metal stents are often used for palliative biliary drainage in these patients, but are currently seen as an absolute contraindication for IRE due to the perceived risk of direct heating of the metal and its surroundings. This study investigates the thermal and tissue viability changes due to a metal stent during IRE.Methods
IRE was performed in a homogeneous tissue model (polyacrylamide gel), without and with a metal stent placed perpendicular and parallel to the electrodes, delivering 90 and 270 pulses (15–35 A, 90 μsec, 1.5 cm active tip exposure, 1.5 cm interelectrode distance, 1000–1500 V/cm, 90 pulses/min), and in-vivo in a porcine liver (4 ablations). Temperature changes were measured with an infrared thermal camera and with fiber-optic probes. Tissue viability after in-vivo IRE was investigated macroscopically using 5-triphenyltetrazolium chloride (TTC) vitality staining.Results
In the gel, direct stent-heating was not observed. Contrarily, the presence of a stent between the electrodes caused a higher increase in median temperature near the electrodes (23.2 vs 13.3°C [90 pulses]; p = 0.021, and 33.1 vs 24.8°C [270 pulses]; p = 0.242). In-vivo, no temperature difference was observed for ablations with and without a stent. Tissue examination showed white coagulation 1mm around the electrodes only. A rim of vital tissue remained around the stent, whereas ablation without stent resulted in complete tissue avitality.Conclusion
IRE in the vicinity of a metal stent does not cause notable direct heating of the metal, but results in higher temperatures around the electrodes and remnant viable tissue. Future studies should determine for which clinical indications IRE in the presence of metal stents is safe and effective. 相似文献13.
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WT Ismaya A Efthyani DS Retnoningrum X Lai BW Dijkstra RR Tjandrawinata 《Biotechnic & histochemistry》2017,92(6):411-416
The light subunit of mushroom, Agaricus bisporus, tyrosinase (LSMT), has been identified as an extrinsic component of the enzyme. Its function is unknown, but it can cross an epithelial cell layer, which suggests that it can be absorbed by the intestine. A similar capability has been demonstrated for the HA-33 component of the progenitor toxin from Clostridium botulinum, which is the closest structural homolog of LSMT. Unlike HA-33, LSMT appears to be non-immunogenic as shown by preliminary tests in Swiss Webster mice. We investigated the immunogenicity and histopathology of LSMT in mice to determine its safety in vivo. LSMT did not evoke generation of antibodies after prolonged periods of intraperitoneal administration. Histopathological observations confirmed the absence of responses in organs after twelve weekly administrations of LSMT. We found that LSMT is not toxic and is less immunogenic than the C. botulinum HA-33 protein, which supports further research and development for pharmaceutical application. 相似文献
15.
Andrew D. Stephens Rachel A. Haggerty Paula A. Vasquez Leandra Vicci Chloe E. Snider Fu Shi Cory Quammen Christopher Mullins Julian Haase Russell M. Taylor II Jolien S. Verdaasdonk Michael R. Falvo Yuan Jin M. Gregory Forest Kerry Bloom 《The Journal of cell biology》2013,200(6):757-772
The mechanisms by which sister chromatids maintain biorientation on the metaphase spindle are critical to the fidelity of chromosome segregation. Active force interplay exists between predominantly extensional microtubule-based spindle forces and restoring forces from chromatin. These forces regulate tension at the kinetochore that silences the spindle assembly checkpoint to ensure faithful chromosome segregation. Depletion of pericentric cohesin or condensin has been shown to increase the mean and variance of spindle length, which have been attributed to a softening of the linear chromatin spring. Models of the spindle apparatus with linear chromatin springs that match spindle dynamics fail to predict the behavior of pericentromeric chromatin in wild-type and mutant spindles. We demonstrate that a nonlinear spring with a threshold extension to switch between spring states predicts asymmetric chromatin stretching observed in vivo. The addition of cross-links between adjacent springs recapitulates coordination between pericentromeres of neighboring chromosomes. 相似文献
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Tianbao Yang Gul Shad Ali Lihua Yang Liqun Du ASN Reddy BW Poovaiah 《Plant signaling & behavior》2010,5(8):991-994
Recently we reported that CRLK1, a novel calcium/calmodulin-regulated receptor-like kinase plays an important role in regulating plant cold tolerance. Calcium/calmodulin binds to CRLK1 and upregulates its activity. Gene knockout and complementation studies revealed that CRLK1 is a positive regulator of plant response to chilling and freezing temperatures. Here we show that MEKK1, a member of MAP kinase kinase kinase family, interacts with CRLK1 both in vitro and in planta. The cold triggered MAP kinase activation in wild-type plants was abolished in crlk1 knockout mutants. Similarly, the cold induced expression levels of genes involved in MAP kinase signaling are also altered in crlk1 mutants. These results suggest that calcium/calmodulinregulated CRLK1 modulates cold acclimation through MAP kinase cascade in plants.Key words: calcium, calmodulin, cold stress, MAPK, Arabidopsis, protein phosphorylationCalcium, a universal second messenger in eukaryotic cells, mediates changes in external and internal signals leading to the physiological responses.1–4 Calcium/calmodulin (Ca2+/CaM)-dependent protein kinases (CaMKs) are very important players in calcium/calmodulin mediated signaling in mammalian cells.5 In plants, Ca2+/CaM-dependent protein phosphorylation was observed more than 25 years ago.6 Several calmodulin-regulated protein kinases have been identified and characterized.7,8 For example, plants have a unique chimeric Ca2+/CaM-dependent protein kinase (CCaMK), which exhibits Ca2+-dependent autophosphorylation and Ca2+/CaM-dependent substrate phosphorylation.9 CCaMK is required for bacterial and fungal symbioses in plants.10–12 Recently, we characterized a novel plant-specific calcium/CaM-regulated receptor-like kinase, CRLK1.13 Ca2+/CaM binds to CRLK1 and stimulates its kinase activity. Functional studies with CRLK1 indicate that CRLK1 acts as a positive regulator in plant response to chilling and freezing temperatures. To further define the CRLK1-mediated signal pathway, we isolated CRLK1 interacting proteins by co-immunoprecipitation using an anti-CRLK1 antibody. Since cold increases the amount of CRLK1 protein, wildtype plants (WT) were treated at 4°C for 1 hr before co-immunoprecipitation. The resulting CRLK1 immunocomplex was separated by SDS-PAGE. We observed several bands of different sizes only in the wild-type but not in the crlk1 knockout mutant plants (Fig. 1A). Furthermore, the intensity of these bands increased upon cold treatment, suggesting that they are the putative partners or associated proteins of the CRLK1 immunocomplex.Open in a separate windowFigure 1CRLK1 Interacts with MEKK1. (A) One-dimension SDS-PAGE of anti-CRLK1 immunocomplexes from 3-week-old WT or crlk1 plants with or without cold treatment. One mg of total protein was used for immunoprecipitation. (B) A list of putative CRLK1-interacting proteins determined by MALDI-TOF-MS analysis. (C) CRLK1 interacts with MEKK1 as shown by GST pull-down assay. (D) BiFC analysis show that CR LK associates with MEKK1 in vivo. Upper row shows that CRLK and MEKK1 associate both on cell membrane and in endosomes. The middle and last rows are controls. Bar = 10 µm.To determine the identities of these proteins, mass spectrometric analysis was performed with the total immunocomplex.14 In addition to CRLK1, there were 12 other proteins which matched the Arabidopsis database. Several of them appeared in the pull-down complex from WT, but not from crlk1 mutants. These putative interacting proteins included MEKK1, another unknown protein kinase, a type 2C phosphatase and CaM (Fig. 1B). MEKK1 is one of the 60 putative MAPKKKs in the Arabidopsis genome, and sits on the top of mitogen-activated protein kinase (MAPK) cascade. The MAPK signaling consists of a cascade of three consecutively acting protein kinases, a MAP kinase kinase kinase (MAPKKK), a MAP kinase kinase (MAPKK) and a MAP kinase (MAPK). Plants possess multiple MAPKKKs, MAPKKs and MAPKs, which respond to different upstream signals and activate distinct downstream pathways.15–17 The specific MAPK module responding to lower temperature has been determined in Arabidopsis.18,19 MEKK1, a member of MAPKKKs, specifically interacts and phosphorylates MKK2 and regulates COR genes expression in response to cold stress.19 MEKK1 has been shown to play a role in mediating reactive oxygen species homeostasis.20,21 Therefore we selected MEKK1 from the putative CRLK1 partners for further studies. 相似文献
18.
Phylogenetic evidence for role-reversals of gender-associated mitochondrial DNA in Mytilus (Bivalvia: Mytilidae) 总被引:1,自引:0,他引:1
Hoeh WR; Stewart DT; Saavedra C; Sutherland BW; Zouros E 《Molecular biology and evolution》1997,14(9):959-967
Distinct gender-associated mitochondrial DNA (mtDNA) lineages (i.e.,
lineages which are transmitted either through males or through females)
have been demonstrated in two families of bivalves, the Mytilidae (marine
mussels) and the Unionidae (freshwater mussels), which have been separated
for more than 400 Myr. The mode of transmission of these M (for
male-transmitted) and F (for female-transmitted) molecules has been
referred to as doubly uniparental inheritance (DUI), in contrast to
standard maternal inheritance (SMI), which is the norm in animals. A
previous study suggested that at least three origins of DUI are required to
explain the phylogenetic pattern of M and F lineages in freshwater and
marine mussels. Here we present phylogenetic evidence based on partial
sequences of the cytochrome c oxidase subunit I gene and the 16S RNA gene
that indicates the DUI is a dynamic phenomenon. Specifically, we
demonstrate that F lineages in three species of Mytilus mussels, M. edulis,
M. trossulus, and M. californianus, have spawned separate lineages which
are now associated only with males. This process is referred to as
"masculinization" of F mtDNA. By extension, we propose that DUI may be a
primitive bivalve character and that periodic masculinization events
combined with extinction of previously existing M types effectively reset
the time of divergence between conspecific gender-associated mtDNA
lineages.
相似文献
19.
Marcel A.G. van der Heyden Paul M.P. Van Bergen en Henegouwen Nancy de Ruiter Marina A.M. Verdaasdonk Jan G. van den Tweel Gert Rijksen Johannes Boonstra Piet Joling 《Experimental cell research》1997,234(2):521
NIH-3T3 fibroblasts expressing epidermal growth factor receptors (EGFRs) lacking the actin binding domain (ABD) were analyzed for their EGF-induced capacity to invade a bone marrow stromal cell (BMSC) monolayer. The fibroblasts display a reduction in the percentage of cytoskeleton-associated EGFRs. Furthermore, EGF-induced tyrosine kinase activity is unaffected by the mutation. Cells expressing the mutant EGFRs hardly invade a BMSC monolayer upon EGF stimulation in contrast to cells expressing wild-type EGFRs. Using the same cells no difference was observed in PDGF-induced invasion, which ligand was as potent in both cell types as EGF was in wild-type cells. Inhibition of both the phosphatidyl inositol-3-kinase (PI-3-K) and lipoxygenase pathways in wild-type cells mimicked the effect of the ABD deletion. Our results point to an important role for the ABD of the EGFR in EGF-induced tissue invasion. 相似文献
20.
DNA fingerprints from hypervariable mitochondrial genotypes 总被引:4,自引:0,他引:4
Conventional surveys of restriction-fragment polymorphisms in mitochondrial
DNA of menhaden fish (Brevoortia tyrannus/patronus complex) and chuckwalla
lizards (Sauromalus obesus) revealed exceptionally high levels of genetic
variation, attributable to differences in mtDNA size as well as in
restriction sites. The observed probabilities that any two randomly drawn
individuals differed detectably in mtDNA genotype were 0.998 and 0.983 in
the two species, respectively. Thus, the variable gel profiles provided
unique mtDNA "fingerprints" for most conspecific animals assayed. mtDNA
fingerprints differ from nuclear DNA fingerprints in several empirical
respects and should find special application in the genetic assessment of
maternity.
相似文献