scholarly journals Ex vivo Induction of Apoptotic Mesenchymal Stem Cell by High Hydrostatic Pressure

2020 ◽  
Author(s):  
Tien Minh Le ◽  
Naoki Morimoto ◽  
Nhung Thi My Ly ◽  
Toshihito Mitsui ◽  
Sharon Claudia Notodihardjo ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
Stromal Cells ◽  
High Hydrostatic Pressure ◽  
Tissue Repair ◽  
Ex Vivo ◽  
Morphological Changes ◽  
Fluid Pressure ◽  
Physical Technique ◽  
Transmission Electron ◽  
The Impact

Abstract Among promising solutions for tissue repair and wound healing, mesenchymal stem (or stromal) cells (MSCs) have been a focus of attention and have become the most clinically studied experimental cell therapy. Recent studies reported the importance of apoptosis in MSC-mediated immunomodulation, in which apoptotic MSCs (apoMSCs) were shown to be superior to living MSCs. Nowadays, high hydrostatic pressure (HHP), a physical technique that uses only fluid pressure, has been developed and applied in various bioscience fields, including biotechnology, biomaterials, and regenerative medicine, as its safe and simply operation. In the current study, we investigated the impact of HHP treatment on human bone marrow-MSC survival and proliferation. Based on the detection of executioner caspase activation, phosphatidylserine exposure, DNA fragmentation (TUNEL) and irrefutable ultrastructural morphological changes on transmission electron microscopy (TEM), our data revealed that HHP treatment induced complete apoptosis in MSCs. Notably, this technique might provide manipulated products for use in cell-based therapies as manufacturing capability expands. We hope that our findings will contribute to the improvement of MSCs or EVs in translational research development.

scholarly journals Ex vivo Inducing Apoptotic Mesenchymal Stem Cell by High Hydrostatic Pressure

2020 ◽  
Author(s):  
Tien Minh Le ◽  
Naoki Morimoto ◽  
Nhung Thi My Ly ◽  
Toshihito Mitsui ◽  
Sharon Claudia Notodihardjo ◽  
...  
Keyword(s):  
Stem Cell ◽  
Hydrostatic Pressure ◽  
Mesenchymal Stem Cell ◽  
High Hydrostatic Pressure ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Morphological Changes ◽  
Fluid Pressure ◽  
Live Cell ◽  
Tunel Staining

Abstract Background: Apoptosis was reported to take crucial role in mesenchymal stem cell (MSC)-mediated immunomodulation, in which apoptotic MSCs were shown to be superior compared to living MSCs. Furthermore, extracellular vesicles (Evs) derived from MSCs were revealed more specific advantages for patient safety such as lower propensity to trigger innate and adaptive immune responses. As a safety and simple operation, high hydrostatic pressure (HHP), a physical technique that uses only fluid pressure to inactivate cells or tissues, has been developed and applied in a lot of field of biosciences, including biotechnology, biomaterials, or tissue engineering. Methods: MSCs isolated from human bone marrow were suspended cultured in appropriate medium and subjected to pressurization at 50 MPa for 36 h. Then cells were collected and investigated apoptotic pathway by transmission electron microscopy (TEM), phosphatidylserine membrane translocations, cleaved caspase-3/7 and terminal deoxy-nucleotidyl transferase dUTP nick-end labeling (TUNEL) staining. Besides, viability assays and live cell imaging were also used for assessement of cell survival after pressurization. Results: We found that HHP at 50 MPa for ≥36 h completely induced MSC death by Live/Dead assay, live cell imaging and WST-8 assay up to 7 days after pressurization. The large amount of apoptotic MSCs death was found based on morphological changes in TEM, phosphatidylserine exposure, caspase activation and detection of DNA fragmentations via TUNEL staining. Conclusions: In the current study, our data revealed that HHP treatment was convenient processing which safety and effectively induced MSCs undergo apoptosis. Especially, by capable of manufacture expanding, this technique might provide numbers of manipulated products using for industrial cell-based therapies.

The impact of high hydrostatic pressure on structure and dynamics of β-lactoglobulin

2013 ◽  
Vol 1830 (10) ◽  
pp. 4974-4980 ◽  
Author(s):  
Daniela Russo ◽  
Maria Grazia Ortore ◽  
Francesco Spinozzi ◽  
Paolo Mariani ◽  
Camille Loupiac ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Structure And Dynamics ◽  
The Impact ◽  
Β Lactoglobulin

Influence of hydrostatic pressure gradients on regulation of plasma volume after exercise

1998 ◽  
Vol 85 (2) ◽  
pp. 667-675 ◽  
Author(s):  
Gary W. Mack ◽  
Roger Yang ◽  
Alan R. Hargens ◽  
Kei Nagashima ◽  
Andrew Haskell
Keyword(s):  
Hydrostatic Pressure ◽  
Osmotic Pressure ◽  
Plasma Volume ◽  
Fluid Pressure ◽  
Colloid Osmotic Pressure ◽  
Upright Position ◽  
Upright Posture ◽  
Immediate Recovery ◽  
Before And After ◽  
The Impact

The impact of posture on the immediate recovery of intravascular fluid and protein after intense exercise was determined in 14 volunteers. Forces which govern fluid and protein movement in muscle interstitial fluid pressure (PISF), interstitial colloid osmotic pressure (COPi), and plasma colloid osmotic pressure (COPp) were measured before and after exercise in the supine or upright position. During exercise, plasma volume (PV) decreased by 5.7 ± 0.7 and 7.0 ± 0.5 ml/kg body weight in the supine and upright posture, respectively. During recovery, PV returned to its baseline value within 30 min regardless of posture. PV fell below this level by 60 and 120 min in the supine and upright posture, respectively ( P < 0.05). Maintenance of PV in the upright position was associated with a decrease in systolic blood pressure, an increase in COPp (from 25 ± 1 to 27 ± 1 mmHg; P < 0.05), and an increase in PISF (from 5 ± 1 to 6 ± 2 mmHg), whereas COPi was unchanged. Increased PISFindicates that the hydrostatic pressure gradient favors fluid movement into the vascular space. However, retention of the recaptured fluid in the plasma is promoted only in the upright posture because of increased COPp.

scholarly journals Identification of Sigma Factor σB-Controlled Genes and Their Impact on Acid Stress, High Hydrostatic Pressure, and Freeze Survival in Listeria monocytogenes EGD-e

2004 ◽  
Vol 70 (6) ◽  
pp. 3457-3466 ◽  
Author(s):  
Henrike H. Wemekamp-Kamphuis ◽  
Jeroen A. Wouters ◽  
Patrick P. L. A. de Leeuw ◽  
Torsten Hain ◽  
Trinad Chakraborty ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Hydrostatic Pressure ◽  
Sigma Factor ◽  
High Hydrostatic Pressure ◽  
Glutamate Decarboxylase ◽  
Expression Profiles ◽  
Pressure Treatment ◽  
Wild Type ◽  
High Hydrostatic Pressure Treatment ◽  
The Impact

ABSTRACT The gene encoding the alternative sigma factor σB in Listeria monocytogenes is induced upon exposure of cells to several stresses. In this study, we investigated the impact of a sigB null mutation on the survival of L. monocytogenes EGD-e at low pH, during high-hydrostatic-pressure treatment, and during freezing. The survival of ΔsigB mutant exponential-phase cells at pH 2.5 was 10,000-fold lower than the survival of EGD-e wild-type cells. Moreover, the ΔsigB mutant failed to show an acid tolerance response. Upon preexposure for 1 h to pH 4.5, the survival at pH 2.5 was 100,000-fold lower for the ΔsigB mutant than for the wild type. The glutamate decarboxylase (GAD) acid resistance system is important in survival and adaptation of L. monocytogenes in acidic conditions. The σB dependence of the gad genes (gadA, gadB, gadC, gadD, and gadE) was analyzed in silico. Putative σB-dependent promoter sites were found upstream of the gadCB operon (encoding a glutamate/γ-aminobutyrate antiporter and a glutamate decarboxylase, respectively) and the lmo2434 gene (gadD, encoding a putative glutamate decarboxylase). Reverse transcriptase PCR revealed that expression of the gadCB operon and expression of gadD are indeed σB dependent. In addition, a proteomics approach was used to analyze the protein expression profiles upon acid exposure. Although the GAD proteins were not recovered, nine proteins accumulated in the wild type but not in the ΔsigB strain. These proteins included Pfk, GalE, ClpP, and Lmo1580. Exposure to pH 4.5, in order to preload cells with active σB and consequently with σ B-dependent general stress proteins, also provided considerable protection against high-hydrostatic-pressure treatment and freezing. The combined data argue that the expression of σB-dependent genes provides L. monocytogenes with nonspecific multiple-stress resistance that may be relevant for survival in the natural environment as well as during food processing.

scholarly journals High Hydrostatic Pressure vs. Thermal Pasteurization: The Effect on the Bioactive Compound Profile of a Citrus Maqui Beverage

Foods ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2416
Author(s):  
Francisco J. Salar ◽  
Paula M. Periago ◽  
Vicente Agulló ◽  
Cristina García-Viguera ◽  
Pablo S. Fernández
Keyword(s):  
Hydrostatic Pressure ◽  
Shelf Life ◽  
High Hydrostatic Pressure ◽  
Storage Period ◽  
Bioactive Compound ◽  
Processing Technique ◽  
Microbial Inactivation ◽  
Processing Technologies ◽  
The Impact ◽  
Thermal Pasteurization

The effects of high hydrostatic pressure (HHP) compared to thermal pasteurization (TP) were studied in healthy citrus-maqui beverages. The impact of the processing technologies on the microbiological and phytochemical profile was assessed by applying two HHP treatments at 450 and 600 MPa for 180 s and TP at 85 °C for 15 s. The shelf life under refrigeration (4 °C) and room temperature (20 °C) was monitored for 90 days. All treatments ensured microbiological stability at both storage temperatures. Aside from that, the physicochemical parameters were not significantly different after processing or throughout the storage period. Regarding color parameters, an increase in the reddish coloration was observed during storage for those beverages treated by HHP. In general, phenolic compounds were little affected by the processing technique, even when treatment under HHP was more stable than by TP during storage. On the other hand, vitamin C showed great degradation after processing under any condition. It can be concluded that HHP is an effective alternative to thermal treatments, achieving effective microbial inactivation and extending the shelf life of the juices by contributing to a better preservation of color and bioactive compounds.

Relationship between Autophagy and Ventilator-induced Diaphragmatic Dysfunction

2015 ◽  
Vol 122 (6) ◽  
pp. 1349-1361 ◽  
Author(s):  
Ilan Azuelos ◽  
Boris Jung ◽  
Martin Picard ◽  
Feng Liang ◽  
Tong Li ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Messenger Rna ◽  
Ex Vivo ◽  
Diaphragm Muscle ◽  
Autophagosome Formation ◽  
Diaphragmatic Dysfunction ◽  
Transmission Electron ◽  
Diaphragmatic Weakness ◽  
The Impact

Abstract Background: Mechanical ventilation (MV) is associated with atrophy and weakness of the diaphragm muscle, a condition termed ventilator-induced diaphragmatic dysfunction (VIDD). Autophagy is a lysosomally mediated proteolytic process that can be activated by oxidative stress, which has the potential to either mitigate or exacerbate VIDD. The primary goals of this study were to (1) determine the effects of MV on autophagy in the diaphragm and (2) evaluate the impact of antioxidant therapy on autophagy induction and MV-induced diaphragmatic weakness. Methods: Mice were assigned to control (CTRL), MV (for 6 h), MV + N-acetylcysteine, MV + rapamycin, and prolonged (48 h) fasting groups. Autophagy was monitored by quantifying (1) autophagic vesicles by transmission electron microscopy, (2) messenger RNA levels of autophagy-related genes, and (3) the autophagosome marker protein LC3B-II, with and without administration of colchicine to calculate the indices of relative autophagosome formation and degradation. Force production by mouse diaphragms was determined ex vivo. Results: Diaphragms exhibited a 2.2-fold (95% CI, 1.8 to 2.5) increase in autophagic vesicles visualized by transmission electron microscopy relative to CTRL after 6 h of MV (n = 5 per group). The autophagosome formation index increased in the diaphragm alone (1.5-fold; 95% CI, 1.3 to 1.8; n = 8 per group) during MV, whereas prolonged fasting induced autophagosome formation in both the diaphragm (2.5-fold; 95% CI, 2.2 to 2.8) and the limb muscle (4.1-fold; 95% CI, 1.8 to 6.5). The antioxidant N-acetylcysteine further augmented the autophagosome formation in the diaphragm during MV (1.4-fold; 95% CI, 1.2 to 1.5; n = 8 per group) and prevented MV-induced diaphragmatic weakness. Treatment with the autophagy-inducing agent rapamycin also largely prevented the diaphragmatic force loss associated with MV (n = 6 per group). Conclusions: In this model of VIDD, autophagy is induced by MV but is not responsible for diaphragmatic weakness. The authors propose that autophagy may instead be a beneficial adaptive response that can potentially be exploited for therapy of VIDD.

scholarly journals Effect of Tannic Acid on the Protective Properties of the in situ Formed Pellicle

2016 ◽  
Vol 51 (1) ◽  
pp. 34-45 ◽  
Author(s):  
Susann Hertel ◽  
Sandra Pötschke ◽  
Sabine Basche ◽  
Judith Delius ◽  
Wiebke Hoth-Hannig ◽  
...  
Keyword(s):  
Tannic Acid ◽  
Ex Vivo ◽  
Oral Exposure ◽  
Dependent Manner ◽  
Pellicle Formation ◽  
Protective Properties ◽  
Transmission Electron ◽  
Bovine Enamel ◽  
The Impact

Objectives: In the present in situ/ex vivo study the impact of tannic acid on the erosion-protective properties of the enamel pellicle was tested. Additionally, the antiadherent and antibacterial effects of tannic acid were evaluated. Methods: The pellicle was formed in situ on bovine enamel samples fixed on individual splints worn by 6 subjects. Following 1 min of pellicle formation the volunteers rinsed for 10 min with tannic acid. After further oral exposure for 19 min, 109 min, and 8 h overnight, respectively, slabs were incubated in HCl ex vivo (pH 2.0, 2.3, 3.0) over 120 s. Subsequently, kinetics of calcium and phosphate release were measured photometrically. Samples after a 1-min fluoride mouth rinse as well as enamel samples with and without a 30-min in situ pellicle served as controls. Antiadherent effects were evaluated after a 1-min rinse with tannic acid and oral exposure of the slabs overnight. DAPI (4′,6-diamidino-2-phenylindole) combined with concanavalin A staining and live/dead staining was used for fluorescence microscopic visualization and quantification of adherent bacteria and glucans. Modification of the pellicle's ultrastructure by tannic acid was evaluated by transmission electron microscopy (TEM). Results: Tannic acid significantly improved the erosion-protective properties of the pellicle in a pH-dependent manner. Bacterial adherence and glucan formation on enamel were significantly reduced after rinses with tannic acid as investigated by fluorescence microscopy. TEM imaging indicated that rinsing with tannic acid yielded a sustainable modification of the pellicle; it was distinctly more electron dense. Conclusion: Tannic acid offers an effective and sustainable approach for the prevention of caries and erosion.

scholarly journals Preliminary evidence that maternal immune activation specifically increases diagonal domain volume in the rat brain during early postnatal development

2018 ◽  
Author(s):  
Tobias C. Wood ◽  
Michelle E. Edye ◽  
Michael K. Harte ◽  
Joanna C. Neill ◽  
Eric P. Prinssen ◽  
...  
Keyword(s):  
Rat Brain ◽  
Immune Activation ◽  
Diffusion Tensor ◽  
Ex Vivo ◽  
Morphological Changes ◽  
Female Offspring ◽  
Preliminary Evidence ◽  
Poly I:C ◽  
Maternal Immune Activation ◽  
The Impact

AbstractMaternal immune activation (MIA) is consistently associated with elevated risk for multiple psychiatric disorders in the affected offspring. Related to this, an important goal of our work is to explore the impact of MIA effects across the lifespan. In this context, we recently reported the effects of poly (I:C)-induced MIA at gestational day (GD)15, immediately prior to birth, at GD21 and again at post-natal day (PD)21, providing a systematic assessment of plasma IL-6, body temperature and weight alterations in pregnant rats following poly (I:C) exposure and preliminary evidence for gross morphological changes and microglial neuropathology in both male and female offspring at GD21 and PD21. Here, we sought to complement and extend these data by characterising in more detail the meso-scale impact of gestational poly (I:C) exposure at GD15 on the neuroanatomy of the juvenile (PD21) rat brain using high-resolution, ex vivo anatomical magnetic resonance imaging (MRI) in combination with atlas-based segmentation. Our preliminary data suggest subtle neuroanatomical effects of gestational poly (I:C) exposure (n=10) relative to saline controls (n=10) at this time-point. Specifically, we report here preliminary evidence for a significant increase in the relative volume of the diagonal domain in poly (I:C) offspring (p<0.01; q<0.1), particularly in female offspring. This occurred in the absence of any microstructural alterations as detectable using diffusion tensor imaging (DTI). Longitudinal in vivo studies, informed by the effect sizes from this dataset are now required to establish both the functional relevance and cellular mechanisms of the apparent DD volume increase.

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