scholarly journals Effects of High Hydrostatic Pressure on Expression Profiles of In Vitro Produced Vitrified Bovine Blastocysts

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Zongliang Jiang ◽  
Patrick Harrington ◽  
Ming Zhang ◽  
Sadie L. Marjani ◽  
Joonghoon Park ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Expression Profiles

scholarly journals 32 EFFECTS OF HIGH HYDROSTATIC PRESSURE ON EXPRESSION PROFILES OF IN VITRO-PRODUCED, VITRIFIED BOVINE BLASTOCYSTS

2016 ◽  
Vol 28 (2) ◽  
pp. 146
Author(s):  
Z. Jiang ◽  
P. Harrington ◽  
M. Zhang ◽  
S. Marjani ◽  
L. Kuo ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Protein Folding ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Developmental Competence ◽  
Significant Difference

High hydrostatic pressure (HHP) has been used to enhance stress tolerance and to promote embryo survival before they are subjected to insulting procedures such as cryopreservation. However, the molecular mechanisms of the beneficial effects of HHP are poorly understood. Here in vitro-produced bovine blastocysts were treated with 40, 60, and 80 MPa of HHP for 1 h at either 25 or 37°C, followed by 3 different recovery periods (0, 1, and 2 h) after HHP before vitrification by the solid surface vitrification method (Dinnyes et al. 2000). The re-expansion rates after vitrification-warming were significantly (P < 0.05) higher in embryos treated with 40 or 60 MPa than controls, demonstrating that HHP promotes the in vitro developmental competence of vitrified bovine embryos. However, 80 MPa resulted in significantly reduced re-expansion rates, suggesting that this pressure started to be lethal to bovine blastocysts. In addition, no significant difference was found on re-expansion rates between 25 and 37°C; data were therefore combined for the 2 temperatures. Microarray analysis revealed a total of 399 differentially expressed transcripts, representing 254 unique genes, among different treatment groups. Gene ontology analysis revealed that HHP at 40 and 60 MPa promoted embryo competence through down-regulation of genes involved in cell death and apoptosis, and up-regulation of RNA processing, cellular growth, and proliferation. Moreover, gene expression was also changed by the length of the recovery time after HHP. The significantly over-represented groups are apoptosis and cell death in the 1-h group, and protein folding, response to unfolded protein, and cell cycle in the 2-h group. Although 80 MPa also up-regulated expression of genes for apoptosis, but it also significantly down-regulated genes for protein folding and cell cycle, which may explain why these embryos stopped developing. Taken together, these data suggest that HHP induces specific responses in vitrified bovine blastocysts and promotes their developmental competence through modest transcriptional reprogramming.

scholarly journals High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification

Reproduction ◽  
2007 ◽  
Vol 135 (1) ◽  
pp. 13-17 ◽  
Author(s):  
Y Du ◽  
C S Pribenszky ◽  
M Molnar ◽  
X Zhang ◽  
H Yang ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Developmental Competence

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.

Verification of the Inactivation of Melanocytic Nevus in vitro Using a Newly Developed Portable High Hydrostatic Pressure Device

2016 ◽  
Vol 201 (3) ◽  
pp. 170-179 ◽  
Author(s):  
Naoki Morimoto ◽  
Chizuru Jinno ◽  
Atsushi Mahara ◽  
Natsuko Kakudo ◽  
Toshia Fujisato ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Physical Method ◽  
Melanocytic Nevus ◽  
Explant Culture ◽  
Cultured Fibroblasts ◽  
Giant Congenital Melanocytic Nevi ◽  
Tissue Specimens ◽  
Congenital Melanocytic Nevi

High hydrostatic pressure (HHP) technology is a physical method for inactivating tissue. We reported that nevus specimens were inactivated after HHP at 200 MPa and that the inactivated nevus could be used as autologous dermis for covering skin defects. In this study, we verified the inactivation of nevus specimens using a newly developed portable HHP device which will be used in a clinical trial. Nevus tissue specimens were obtained from 5 patients (mean age 7.2 years, range 1-19). We cultured fibroblasts and nevus cells from the tissue specimens and then evaluated their inactivation after HHP at 200 MPa by confirming the attachment of the suspensions and by the live/dead staining of the suspensions, through the dissociation of the cells on chamber slides and by the live/dead staining of the remaining cells. The cells were also quantitatively evaluated by WST-8 assay. We then confirmed the inactivation of the nevus specimens after HHP using explant culture. Our results indicated that fibroblasts and nevus cells were inactivated after HHP at 200 MPa, with the exception of a small percentage of green-colored cells, which reflected the remaining activity of the cellular esterases after HHP. No cells migrated from the nevus specimens after HHP at 200 MPa. We verified the inactivation of fibroblasts and nevus cells cultured from nevus specimens, and in the nevus samples themselves after pressurization at 200 MPa using this device. This device could be used in clinical trials for giant congenital melanocytic nevi and may thus become useful in various medical fields.

High hydrostatic pressure influences the in vitro response to xenobiotics in Dicentrarchus labrax liver

2016 ◽  
Vol 173 ◽  
pp. 43-52 ◽  
Author(s):  
Benjamin Lemaire ◽  
Eric Mignolet ◽  
Cathy Debier ◽  
Pedro Buc Calderon ◽  
Jean Pierre Thomé ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Dicentrarchus Labrax ◽  
In Vitro Response

28 HIGH HYDROSTATIC PRESSURE IMPROVED DEVELOPMENTAL COMPETENCE OF PORCINE OOCYTES FOR HANDMADE CLONING

2008 ◽  
Vol 20 (1) ◽  
pp. 94 ◽  
Author(s):  
Y. Du ◽  
L. Lin ◽  
C. Pribenszky ◽  
M. Molnár ◽  
P. M. Kragh ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Nuclear Transfer ◽  
Technical Assistance ◽  
Developmental Competence ◽  
Control Groups ◽  
Donor Cells ◽  
And Control ◽  
Handmade Cloning

High hydrostatic pressure (HHP) has been introduced into the field of embryology recently, with the possible mechanism that a sublethal HHP could induce the synthesis of molecular chaperons to protect the embryos from further stresses. Improved cryotolerance has been achieved successfully in HHP-treated mouse (Pribenszky 2005 Anim. Reprod. Sci. 87, 143–150) and bovine (Pribenszky 2005 Reprod. Domest. Anim. 40, 338) embryos, and the semen of bull (Pribenszky 2007 Reprod. Fertil. Dev. 19, 181–182) and boar (Pribenszky 2005 Reprod. Fertil. Dev. 18, 162–163). The objective of the present study was to apply this new technique to in vitro-matured (IVM) porcine oocytes and further investigate its effect in the procedure of handmade cloning (HMC). After 40 h IVM, cumulus–oocyte complexes (COCs) were loaded in 0.5-mL straws by a 2-mL syringe, with HEPES-buffered TCM199 as the loading medium. COCs were then treated with 20 MPa (200 times greater than atmospheric pressure) for 60 min by a pressurizing device (Cryo-Innovation Inc., Budapest, Hungary), with an interval of 120 min between HHP treatment and subsequent HMC. Two different cell lines (from Day 40 fetuses of Yucatan and Danish Landrace breeds (LW1-2)) were used as donor cells for nuclear transfer. A total of 592 reconstructed embryos were produced from both HHP-treated and control groups and were in vitro cultured for 6 days to evaluate the developmental competence through to blastocyst formation. The effect of donor cells on blastocyst development was also investigated. SPSS 11.0 program (SPSS, Inc., Chicago, IL, USA) was used for statistical analysis; values with P < 0.05 were regarded as significant. Blastocyst rates of the different groups are shown in Table 1. Our results indicated that COCs treated with HHP had a much higher blastocyst rate than those untreated (P < 0.01) and this improvement was not affected by using different donor cells for nuclear transfer. In conclusion, the sublethal HHP treatment could improve the in vitro developmental competence of porcine IVM oocytes when they are used for HMC. Further in vivo experiments are required to investigate the long-term effect of HHP on embryo development. Table 1. Day 6 blastocyst rates of HHP-treated and control groups with different donor cells for nuclear transfer The authors thank Ruth Kristensen and Janne Adamsen for their help and excellent technical assistance.

Effects of high hydrostatic pressure on bacterial cytoskeleton FtsZ polymers in vivo and in vitro

Microbiology ◽  
2004 ◽  
Vol 150 (6) ◽  
pp. 1965-1972 ◽  
Author(s):  
Akihiro Ishii ◽  
Takako Sato ◽  
Masaaki Wachi ◽  
Kazuo Nagai ◽  
Chiaki Kato
Keyword(s):  
Cell Division ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Elevated Pressure ◽  
Immunofluorescent Staining ◽  
E Coli ◽  
Division Process ◽  
Indirect Immunofluorescent

Some rod-shaped bacteria, including Escherichia coli, exhibit cell filamentation without septum formation under high-hydrostatic-pressure conditions, indicating that the cell-division process is affected by hydrostatic pressure. The effects of elevated pressure on FtsZ-ring formation in E. coli cells were examined using indirect immunofluorescence microscopy. Elevated pressure of 40 MPa completely inhibited colony formation of E. coli cells under the cultivation conditions used, and the cells exhibited obviously filamentous shapes. In the elongated cells, normal cell-division processes appeared to be inhibited, because no FtsZ rings were observed by indirect immunofluorescent staining. In addition, it was observed that hydrostatic pressure dissociated the E. coli FtsZ polymers in vitro. These results suggest that high hydrostatic pressure directly affects cell survival and morphology through the dissociation of the cytoskeletal frameworks.

scholarly journals High Hydrostatic Pressure for Disinfection of Bone Grafts and Biomaterials: An Experimental Study

2009 ◽  
Vol 3 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Hans Gollwitzer ◽  
Wolfram Mittelmeier ◽  
Monika Brendle ◽  
Patrick Weber ◽  
Thomas Miethke ◽  
...  
Keyword(s):  
Hydrostatic Pressure ◽  
Bacterial Growth ◽  
High Hydrostatic Pressure ◽  
Chronic Osteomyelitis ◽  
Bone Grafts ◽  
Biological Properties ◽  
Human Bone ◽  
Bacterial Inactivation ◽  
Bacterial Survival

Background:Autoclaving, heat, irradiation or chemical detergents are used to disinfect autografts, allografts and biomaterials for tissue reconstruction. These methods are often associated with deterioration of mechanical, physical, and biological properties of the bone grafts and synthetic implants. High hydrostatic pressure has been proposed as a novel method preserving biomechanical and biological properties of bone, tendon and cartilage. This is the first study to assess the inactivation of clinically relevant bacteria on biomaterials and human bone by high hydrostatic pressure.Methods:Bacterial suspensions ofStaphylococcus aureus,Pseudomonas aeruginosaandEnterococcus faecium, implants covered with infected blood, human bone infectedin vitro, and biopsies of patients with chronic osteomyelitis were subjected to different protocols of high hydrostatic pressure up to 600 MPa. Bacterial survival after high hydrostatic pressure treatment was determined and compared with bacterial growth in untreated controls.Results:S. aureusandP. aeruginosain suspension were completely inactivated by high hydrostatic pressure (> 5log levels), whereasE. faeciumshowed barotolerance up to 600 MPa. Blood and adherence to metal implants did not significantly alter inactivation of bacteria, and complete disinfection was achieved with barotolerant bacteria (S. aureusandP. aeruginosa). However, osteoarthritic bone demonstrated a non-homogeneous baroprotective effect, with single bone samples resistant to treatment resulting in unaltered bacterial growth, and complete disinfection of artificially infected bone specimens was achieved in 66% forS. aureus, 60% forP. aeruginosaand 0% forE. faecium. Human bone samples of patients with chronic osteomyelitis could be completely disinfected in 2 of 37 cases.Conclusion:High hydrostatic pressure offers new perspectives for disinfection of sensitive biomaterials and bone grafts, and contamination by blood did not significantly affect bacterial inactivation rates. However, a significant baroprotective effect was demonstrated in bone. Effectiveness is currently limited to colonization and / or infection with barosensitive micro-organisms.

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