High concentration of trichlorfon (1 mM) disrupts axonal cytoskeleton and decreases the expression of plasticity-related proteins in SH-SY5Y cells

Reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) can exert opposed effects depending on the dosage: low levels can be involved in signalling and adaptive processes, while higher levels can exert deleterious effects in cells and tissues. Our aim was to emulate a chronic ex vivo oxidative stress situation through a 2 h exposure of immune cells to sustained H2O2 produced by glucose oxidase (GOX), at high or low production rate, in order to determine dissimilar responses of peripheral blood mononuclear cells (PBMCs) and neutrophils on ROS and cytokine production, and mitochondrial dynamics-related proteins, pro/anti-inflammatory and anti-oxidant gene expression. Immune cells were obtained from subjects with metabolic syndrome. H2O2 at low concentrations can trigger a transient anti-inflammatory adiponectin secretion and reduced gene expression of toll-like receptors (TLRs) in PBMCs but may act as a stimulator of proinflammatory genes (IL6, IL8) and mitochondrial dynamics-related proteins (Mtf2, NRF2, Tfam). H2O2 at a high concentration enhances the expression of pro-inflammatory genes (TLR2 and IL1β) and diminishes the expression of mitochondrial dynamics-related proteins (Mtf1, Tfam) and antioxidant enzymes (Cu/Zn SOD) in PBMCs. The GOX treatments produce dissimilar changes in immune cells: Neutrophils were more resistant to H2O2 effects and exhibited a more constant response in terms of gene expression than PBMCs. We observe emerging roles of H2O2 in mitochondrial dynamics and redox and inflammation processes in immune cells.

Download Full-text

High-Efficient Production of Adipose-Derived Stem Cell (ADSC) Secretome Through Maturation Process and Its Non-scarring Wound Healing Applications

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2021.681501 ◽

2021 ◽

Vol 9 ◽

Author(s):

Young-Hyeon An ◽

Dae Hyun Kim ◽

Eun Jung Lee ◽

Dabin Lee ◽

Mihn Jeong Park ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Tissue Repair ◽

High Dose ◽

Efficient Production ◽

Skin Defect ◽

High Concentration ◽

Tissue Repair And Regeneration ◽

High Efficient ◽

Related Proteins

Recently, the stem cell-derived secretome, which is the set of proteins expressed by stem cells and secreted into the extracellular space, has been demonstrated as a critical contributor for tissue repair. In this study, we have produced two sets of high concentration secretomes from adipose-derived mesenchymal stem cells (ADSCs) that contain bovine serum or free of exogenous molecules. Through proteomic analysis, we elucidated that proteins related to extracellular matrix organization and growth factor-related proteins are highly secreted by ADSCs. Additionally, the application of ADSC secretome to full skin defect showed accelerated wound closure, enhanced angiogenic response, and complete regeneration of epithelial gaps. Furthermore, the ADSC secretome was capable of reducing scar formation. Finally, we show high-dose injection of ADSC secretome via intraperitoneal or transdermal delivery demonstrated no detectable pathological conditions in various tissues/organs, which supports the notion that ADSC secretome can be safely utilized for tissue repair and regeneration.

Download Full-text

The Activation of ROS/NF-κB/MMP-9 Pathway Promotes Calcium-Induced Kidney Crystal Deposition

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/8836355 ◽

2021 ◽

Vol 2021 ◽

pp. 1-20

Author(s):

Yue Wu ◽

Jiaqiao Zhang ◽

Cong Li ◽

Henglong Hu ◽

Baolong Qin ◽

...

Keyword(s):

Stone Formation ◽

Rat Kidney ◽

Idiopathic Hypercalciuria ◽

Renal Tubules ◽

Sprague Dawley ◽

High Concentration ◽

Crystal Deposition ◽

Ectopic Tissue ◽

High Calcium ◽

Related Proteins

Idiopathic hypercalciuria is an important risk factor for the formation of calcium-containing kidney stones. Matrix metalloproteinase-9 (MMP-9) is closely related to cell and tissue remodeling and is involved in ectopic tissue calcification. However, little is known about its role in kidney stone formation. In this study, we found that the expression of MMP-9 and that of osteoblastic-related proteins was increased in normal rat kidney epithelial-like (NRK-52E) cells following treatment with a high concentration of calcium, while the knockout or overexpression of MMP-9 could, respectively, significantly inhibit or upregulate the expression of osteoblastic-related proteins and calcium crystal deposition. In addition, apoptosis and calcium crystal deposition were significantly reduced in Sprague–Dawley rats with 1,25(OH)2D3-induced hypercalciuria following MMP-9 inhibitor I treatment. Furthermore, inhibiting reactive oxygen species (ROS) production or the nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) pathway significantly reduced calcium-induced MMP-9 expression and calcium crystal deposition. In summary, our results suggested that a high calcium concentration promotes epithelial–osteoblastic transformation and calcium crystal deposition in renal tubule cells by regulating the ROS/NF-κB/MMP-9 axis and identified a novel role for MMP-9 in regulating calcium-induced calcium crystal deposition in renal tubules.

Download Full-text

Cytomembrane Preservation in Tissue Dehydrated Only With Glutaraldehyde and Epon 812 Resin

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100072046 ◽

1973 ◽

Vol 31 ◽

pp. 320-321

Author(s):

Daniel C. Pease

Keyword(s):

Diffraction Pattern ◽

X Ray Diffraction ◽

High Concentration ◽

Unpublished Study ◽

X Ray ◽

Sectioned Material

A previous study demonstrated that tissue could be successfully infiltrated with 50% glutaraldehyde, and then subsequently polymerized with urea to create an embedment which retained cytomembrane lipids in sectioned material. As a result, the 180-190 Å periodicity characteristic of fresh, mammalian myelin was preserved in sections, as was a brilliant birefringence, and the capacity to bind OsO4 vapor in the hydrophobic bilayers. An associated (unpublished) study, carried out in co-operation with Drs. C.K. Akers and D.F. Parsons, demonstrated that the high concentration of glutaraldehyde (and urea) did not significantly alter the X-ray diffraction pattern of aldehyde-fixed, myelin. Thus, by itself, 50% glutaraldehyde has little effect upon cytomembrane systems and can be used with confidence for the first stages of dehydration.

Download Full-text

Effects of carbamylcholine on chick embryo aggregate retina cell cultures

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100103814 ◽

1988 ◽

Vol 46 ◽

pp. 350-351

Author(s):

Glenn M. Cohen ◽

Radharaman Ray

Keyword(s):

Cell Cultures ◽

Single Cells ◽

Retinal Cell ◽

Cell Aggregates ◽

High Concentration ◽

In Ovo ◽

Sterile Culture ◽

Retinal Tissue ◽

Synaptic Junctions ◽

And Control

Retinal,cell aggregates develop in culture in a pattern similar to the in ovo retina, forming neurites first and then synapses. In the present study, we continuously exposed chick retinal cell aggregates to a high concentration (1 mM) of carbamylcholine (carbachol), an acetylcholine (ACh) analog that resists hydrolysis by acetylcholinesterase (AChE). This situation is similar to organophosphorus anticholinesterase poisoning in which the ACh level is elevated at synaptic junctions due to inhibition of AChE, Our objective was to determine whether continuous carbachol exposure either damaged cholino- ceptive neurites, cell bodies, and synaptic elements of the aggregates or influenced (hastened or retarded) their development.The retinal tissue was isolated aseptically from 11 day embryonic White Leghorn chicks and then enzymatically (trypsin) and mechanically (trituration) dissociated into single cells. After washing the cells by repeated suspension and low (about 200 x G) centrifugation twice, aggregate cell cultures (about l0 cells/culture) were initiated in 1.5 ml medium (BME, GIBCO) in 35 mm sterile culture dishes and maintained as experimental (containing 10-3 M carbachol) and control specimens.

Download Full-text

The Fine Structure of Phloem Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100119867 ◽

1985 ◽

Vol 43 ◽

pp. 632-635

Author(s):

James Cronshaw

Keyword(s):

Structural Studies ◽

High Concentration ◽

Long Distance ◽

Phloem Tissue ◽

Sieve Elements ◽

Long Distance Transport ◽

P Protein ◽

Companion Cells ◽

Electron Microscopical ◽

Distance Transport

Long distance transport in plants takes place in phloem tissue which has characteristic cells, the sieve elements. At maturity these cells have sieve areas in their end walls with specialized perforations. They are associated with companion cells, parenchyma cells, and in some species, with transfer cells. The protoplast of the functioning sieve element contains a high concentration of sugar, and consequently a high hydrostatic pressure, which makes it extremely difficult to fix mature sieve elements for electron microscopical observation without the formation of surge artifacts. Despite many structural studies which have attempted to prevent surge artifacts, several features of mature sieve elements, such as the distribution of P-protein and the nature of the contents of the sieve area pores, remain controversial.

Download Full-text

Tem analysis of multiple-energy arsenic implanted and Rta'd silicon

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100126135 ◽

1987 ◽

Vol 45 ◽

pp. 246-247

Author(s):

R.A. Herring

Keyword(s):

Device Fabrication ◽

High Concentration ◽

Tem Analysis ◽

Defect Clusters ◽

High Fluences ◽

Small Defect ◽

Before And After ◽

The Matrix ◽

The Difference ◽

Factor Type

Rapid thermal annealing (RTA) of ion-implanted Si is important for device fabrication. The defect structures of 2.5, 4.0, and 6.0 MeV As-implanted silicon irradiated to fluences of 2E14, 4E14, and 6E14, respectively, have been analyzed by electron diffraction both before and after RTA at 1100°C for 10 seconds. At such high fluences and energies the implanted As ions change the Si from crystalline to amorphous. Three distinct amorphous regions emerge due to the three implantation energies used (Fig. 1). The amorphous regions are separated from each other by crystalline Si (marked L1, L2, and L3 in Fig. 1) which contains a high concentration of small defect clusters. The small defect clusters were similar to what had been determined earlier as being amorphous zones since their contrast was principally of the structure-factor type that arises due to the difference in extinction distance between the matrix and damage regions.

Download Full-text

Influence of nascent polypeptide positive charges on translation dynamics

Biochemical Journal ◽

10.1042/bcj20200303 ◽

2020 ◽

Vol 477 (15) ◽

pp. 2921-2934

Author(s):

Rodrigo D. Requião ◽

Géssica C. Barros ◽

Tatiana Domitrovic ◽

Fernando L. Palhano

Keyword(s):

Mrna Decay ◽

Translation Termination ◽

Published Data ◽

Translation Efficiency ◽

Amino Acid Residues ◽

High Concentration ◽

Strong Argument ◽

Translation Arrest ◽

Exit Tunnel ◽

Positively Charged

Protein segments with a high concentration of positively charged amino acid residues are often used in reporter constructs designed to activate ribosomal mRNA/protein decay pathways, such as those involving nonstop mRNA decay (NSD), no-go mRNA decay (NGD) and the ribosome quality control (RQC) complex. It has been proposed that the electrostatic interaction of the positively charged nascent peptide with the negatively charged ribosomal exit tunnel leads to translation arrest. When stalled long enough, the translation process is terminated with the degradation of the transcript and an incomplete protein. Although early experiments made a strong argument for this mechanism, other features associated with positively charged reporters, such as codon bias and mRNA and protein structure, have emerged as potent inducers of ribosome stalling. We carefully reviewed the published data on the protein and mRNA expression of artificial constructs with diverse compositions as assessed in different organisms. We concluded that, although polybasic sequences generally lead to lower translation efficiency, it appears that an aggravating factor, such as a nonoptimal codon composition, is necessary to cause translation termination events.

Download Full-text