scholarly journals Conservation of endo-glucanase 16 (EG16) activity across highly divergent plant lineages

2021 ◽  
Author(s):  
Hila Behar ◽  
Kazune Tamura ◽  
Edward R. Wagner ◽  
Daniel Cosgrove ◽  
Harry Brumer
Keyword(s):  
Physcomitrella Patens ◽  
Ex Vivo ◽  
Evolutionary Relationship ◽  
Distinct Group ◽  
Plant Evolution ◽  
Beta Glucan ◽  
Genome Survey ◽  
Sister Clade ◽  
Dynamic Structures

Plant cell walls are highly dynamic structures that are composed predominately of polysaccharides. As such, endogenous carbohydrate active-enzymes (CAZymes) are central to the synthesis and subsequent modification of plant cells during morphogenesis. The endo-glucanase 16 (EG16) members constitute a distinct group of plant CAZymes, angiosperm orthologs of which were recently shown to have dual beta-glucan/xyloglucan hydrolase activity.  Molecular phylogeny indicates that EG16 members comprise a sister clade with a deep evolutionary relationship to the widely studied apoplastic xyloglucan endo-transglycosylases/hydrolases (XTH).  A cross-genome survey indicated that EG16 members occur as a single ortholog across species and are widespread in early-diverging plants, including the non-vascular bryophytes, for which functional data were previously lacking. Remarkably, enzymological characterization of an EG16 ortholog from the model moss Physcomitrella patens (PpEG16) revealed that EG16 activity and sequence/structure are highly conserved across 500 million years of plant evolution, vis-à-vis orthologs from grapevine and poplar.  Ex vivo biomechanical assays demonstrated that the application of EG16 gene products caused abrupt breakage of etiolated hypocotyls rather than slow extension, thereby indicating a mode-of-action distinct from endogenous expansins and microbial endo-glucanases.  The biochemical data presented here will inform future genomic, genetic, and physiological studies of these enzymes.

Efficacy of single-source rapid kV-switching dual-energy CT for characterization of non-uric acid renal stones: a prospective ex vivo study using anthropomorphic phantom

2019 ◽  
Vol 45 (4) ◽  
pp. 1092-1099
Author(s):  
Roberto Cannella ◽  
Mohammed Shahait ◽  
Alessandro Furlan ◽  
Feng Zhang ◽  
Joel D. Bigley ◽  
...  
Keyword(s):  
Uric Acid ◽  
Ex Vivo ◽  
Renal Stones ◽  
Dual Energy ◽  
Dual Energy Ct ◽  
Single Source ◽  
Anthropomorphic Phantom ◽  
Ex Vivo Study

scholarly journals Morpho-molecular ex vivo detection and grading of non-muscle-invasive bladder cancer using forward imaging probe based multimodal optical coherence tomography and Raman spectroscopy

The Analyst ◽  
2020 ◽  
Vol 145 (4) ◽  
pp. 1445-1456 ◽  
Author(s):  
Fabian Placzek ◽  
Eliana Cordero Bautista ◽  
Simon Kretschmer ◽  
Lara M. Wurster ◽  
Florian Knorr ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Optical Coherence Tomography ◽  
Imaging System ◽  
Ex Vivo ◽  
Large Field ◽  
Optical Coherence ◽  
Fiber Optic Probe ◽  
Optic Probe ◽  
Muscle Invasive

Characterization of bladder biopsies, using a combined fiber optic probe-based optical coherence tomography and Raman spectroscopy imaging system that allows a large field-of-view imaging and detection and grading of cancerous bladder lesions.

scholarly journals Consequences of SUR2[A478V] Mutation in Skeletal Muscle of Murine Model of Cantu Syndrome

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1791
Author(s):  
Rosa Scala ◽  
Fatima Maqoud ◽  
Nicola Zizzo ◽  
Giuseppe Passantino ◽  
Antonietta Mele ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Katp Channel ◽  
Ex Vivo ◽  
Channel Modulation ◽  
Flexor Digitorum Brevis ◽  
Inflammatory Edema ◽  
Flexor Digitorum ◽  
Channel Currents

(1) Background: Cantu syndrome (CS) arises from gain-of-function (GOF) mutations in the ABCC9 and KCNJ8 genes, which encode ATP-sensitive K+ (KATP) channel subunits SUR2 and Kir6.1, respectively. Most CS patients have mutations in SUR2, the major component of skeletal muscle KATP, but the consequences of SUR2 GOF in skeletal muscle are unknown. (2) Methods: We performed in vivo and ex vivo characterization of skeletal muscle in heterozygous SUR2[A478V] (SUR2wt/AV) and homozygous SUR2[A478V] (SUR2AV/AV) CS mice. (3) Results: In SUR2wt/AV and SUR2AV/AV mice, forelimb strength and diaphragm amplitude movement were reduced; muscle echodensity was enhanced. KATP channel currents recorded in Flexor digitorum brevis fibers showed reduced MgATP-sensitivity in SUR2wt/AV, dramatically so in SUR2AV/AV mice; IC50 for MgATP inhibition of KATP currents were 1.9 ± 0.5 × 10−5 M in SUR2wt/AV and 8.6 ± 0.4 × 10−6 M in WT mice and was not measurable in SUR2AV/AV. A slight rightward shift of sensitivity to inhibition by glibenclamide was detected in SUR2AV/AV mice. Histopathological and qPCR analysis revealed atrophy of soleus and tibialis anterior muscles and up-regulation of atrogin-1 and MuRF1 mRNA in CS mice. (4) Conclusions: SUR2[A478V] “knock-in” mutation in mice impairs KATP channel modulation by MgATP, markedly so in SUR2AV/AV, with atrophy and non-inflammatory edema in different skeletal muscle phenotypes.

scholarly journals Design of a force-measuring setup for colorectal compression anastomosis and first ex-vivo results

2021 ◽  
Author(s):  
Jana Steger ◽  
Isabella Patzke ◽  
Maximilian Berlet ◽  
Stefanie Ficht ◽  
Markus Eblenkamp ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Tissue Fixation ◽  
Force Transmission ◽  
Tissue Thickness ◽  
Colon Tissue ◽  
Device Development ◽  
Compression Anastomosis ◽  
Technical Requirements ◽  
Significant Difference

Abstract Purpose The introduction of novel endoscopic instruments is essential to reduce trauma in visceral surgery. However, endoscopic device development is hampered by challenges in respecting the dimensional restrictions, due to the narrow access route, and by achieving adequate force transmission. As the overall goal of our research is the development of a patient adaptable, endoscopic anastomosis manipulator, biomechanical and size-related characterization of gastrointestinal organs are needed to determine technical requirements and thresholds to define functional design and load-compatible dimensioning of devices. Methods We built an experimental setup to measure colon tissue compression piercing forces. We tested 54 parameter sets, including variations of three tissue fixation configurations, three piercing body configurations (four, eight, twelve spikes) and insertion trajectories of constant velocities (5 mms−1, 10 mms−1,15 mms−1) and constant accelerations (5 mms−2, 10 mms−2, 15 mms−2) each in 5 samples. Furthermore, anatomical parameters (lumen diameter, tissue thickness) were recorded. Results There was no statistically significant difference in insertion forces neither between the trajectory groups, nor for variation of tissue fixation configurations. However, we observed a statistically significant increase in insertion forces for increasing number of spikes. The maximum mean peak forces for four, eight and twelve spikes were 6.4 ± 1.5 N, 13.6 ± 1.4 N and 21.7 ± 5.8 N, respectively. The 5th percentile of specimen lumen diameters and pierced tissue thickness were 24.1 mm and 2.8 mm, and the 95th percentiles 40.1 mm and 4.8 mm, respectively. Conclusion The setup enabled reliable biomechanical characterization of colon material, on the base of which design specifications for an endoscopic anastomosis device were derived. The axial implant closure unit must enable axial force transmission of at least 28 N (22 ± 6 N). Implant and applicator diameters must cover a range between 24 and 40 mm, and the implant gap, compressing anastomosed tissue, between 2 and 5 mm.

scholarly journals Characterization of Intrinsically Radiolabeled Poly(lactic-co-glycolic acid) Nanoparticles for ex Vivo Autologous Cell Labeling and in Vivo Tracking

2021 ◽  
Author(s):  
Massis Krekorian ◽  
Gerwin G. W. Sandker ◽  
Kimberley R. G. Cortenbach ◽  
Oya Tagit ◽  
N. Koen van Riessen ◽  
...  
Keyword(s):  
Glycolic Acid ◽  
Ex Vivo ◽  
Cell Labeling ◽  
Autologous Cell

scholarly journals Peculiar Evolutionary History of miR390-Guided TAS3-Like Genes in Land Plants

2013 ◽  
Vol 2013 ◽  
pp. 1-14 ◽  
Author(s):  
Maria S. Krasnikova ◽  
Denis V. Goryunov ◽  
Alexey V. Troitsky ◽  
Andrey G. Solovyev ◽  
Lydmila V. Ozerova ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Physcomitrella Patens ◽  
Land Plant ◽  
Plant Evolution ◽  
Marchantia Polymorpha ◽  
History Of ◽  
Molecular Machinery ◽  
Specific Mrnas ◽  
Land Plant Evolution ◽  
Phylogenetic Profiling

PCR-based approach was used as a phylogenetic profiling tool to probe genomic DNA samples from representatives of evolutionary distant moss taxa, namely, classes Bryopsida, Tetraphidopsida, Polytrichopsida, Andreaeopsida, and Sphagnopsida. We found relatives of allPhyscomitrella patensmiR390 and TAS3-like loci in these plant taxa excluding Sphagnopsida. Importantly, cloning and sequencing ofMarchantia polymorphagenomic DNA showed miR390 and TAS3-like sequences which were also found among genomic reads ofM. polymorphaat NCBI database. Our data suggest that the ancient plant miR390-dependent TAS molecular machinery firstly evolved to target AP2-like mRNAs in Marchantiophyta and only then both ARF- and AP2-specific mRNAs in mosses. The presented analysis shows that moss TAS3 families may undergone losses of tasiAP2 sites during evolution toward ferns and seed plants. These data confirm that miR390-guided genes coding for ARF- and AP2-specific ta-siRNAs have been gradually changed during land plant evolution.

About Puncture Testing Applied for Mechanical Characterization of Fetal Membranes

2014 ◽  
Vol 136 (11) ◽  
Author(s):  
Wilfried Bürzle ◽  
Edoardo Mazza ◽  
John J. Moore
Keyword(s):  
Deformation Behavior ◽  
Mechanical Characterization ◽  
Ex Vivo ◽  
Fetal Membrane ◽  
Membrane Rupture ◽  
Resistance To Deformation ◽  
Ex Vivo Testing ◽  
Insight Into

Puncture testing has been applied in several studies for the mechanical characterization of human fetal membrane (FM) tissue, and significant knowledge has been gained from these investigations. When comparing results of mechanical testing (puncture, inflation, and uniaxial tension), we have observed discrepancies in the rupture sequence of FM tissue and significant differences in the deformation behavior. This study was undertaken to clarify these discrepancies. Puncture experiments on FM samples were performed to reproduce previous findings, and numerical simulations were carried out to rationalize particular aspects of membrane failure. The results demonstrate that both rupture sequence and resistance to deformation depend on the samples' fixation. Soft fixation leads to slippage in the clamping, which reduces mechanical loading of the amnion layer and results in chorion rupturing first. Conversely, the stiffer, stronger, and less extensible amnion layer fails first if tight fixation is used. The results provide a novel insight into the interpretation of ex vivo testing as well as in vivo membrane rupture.

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