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.

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

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.

Combination of preoperative embolization and surgery in the treatment of a giant congenital neuroid nevus of the forehead and scalp in a child

2009 ◽  
Vol 3 (3) ◽  
pp. 215-219
Author(s):  
Alexander Margulis ◽  
Benjamin Z. Koplewitz ◽  
Alexander Maly ◽  
José Cohen ◽  
Ravit Yanko-Arzi ◽  
...  
Keyword(s):  
Male Infant ◽  
Large Mass ◽  
Melanocytic Nevus ◽  
Preoperative Embolization ◽  
Congenital Melanocytic Nevus ◽  
Complete Excision ◽  
Melanocytic Nevi ◽  
Giant Congenital Melanocytic Nevi ◽  
Congenital Melanocytic Nevi ◽  
Staged Reconstruction

Congenital melanocytic nevi of neuroectodermal origin are composed primarily of melanocytes and occasional neural elements. A subset of large/giant congenital melanocytic nevi is characterized by neuroidal differentiation resembling the histological pattern of neurofibroma (neuroid congenital melanocytic nevi). The authors report the case of a male infant born with a neuroid congenital melanocytic nevus presenting as a large mass covering extensive portions of his scalp and forehead. The atypical feeding arteries to the nevus were larger in diameter then the patient's internal carotid arteries and formed a rich network of large blood vessels in the nevus. Selective preoperative embolization using calibrated particles was followed by a complete excision and staged reconstruction with tissue expanders. The authors believe that this multidisciplinary approach allowed for a safe surgical procedure with minimal blood loss for this most unusual pediatric head and neck tumor.

scholarly journals Giant Congenital Melanocytic Nevi and Neurocutaneous Melanosis

2015 ◽  
Vol 2015 ◽  
pp. 1-5 ◽  
Author(s):  
Catarina Araújo ◽  
Cristina Resende ◽  
Francisco Pardal ◽  
Celeste Brito
Keyword(s):  
Multiorgan Failure ◽  
Melanocytic Nevus ◽  
Congenital Melanocytic Nevus ◽  
Cerebral Magnetic Resonance Imaging ◽  
Neurocutaneous Melanosis ◽  
Melanocytic Nevi ◽  
Giant Congenital Melanocytic Nevi ◽  
Congenital Melanocytic Nevi ◽  
Leptomeningeal Infiltration ◽  
Expansive Lesion

Introduction. The major medical concern with giant congenital melanocytic nevi CMN is high risk of developing cutaneous melanoma, leptomeningeal melanoma, and neurocutaneous melanocytosis.Case Report. A 30-year-old woman with a giant congenital melanocytic nevus covering nearly the entire right thoracodorsal region and multiple disseminated melanocytic nevi presented with neurological symptoms. Cerebral magnetic resonance imaging revealed a large expansive lesion in the left frontal region. Postsurgically pathological diagnosis revealed characteristics of melanoma. Immunohistochemical examination showed S100(+), HMB45(+), MelanA(+), and MiTF(+). She received radiotherapy with temozolomide followed by two more chemotherapy cycles with temozolomide. She followed a rapidly progressive course, reflecting widespread leptomeningeal infiltration, and she died of multiorgan failure seven months after diagnosis of cerebral melanoma.Discussion. This patient was diagnosed as having a neurocutaneous melanosis with malignant widespread leptomeningeal infiltration. Diffuse spinal involvement is unusual and is described in only another patient.

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