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Sample records for amnion

  1. Staged closure of a giant omphalocele with amnion preservation, modified technique

    Directory of Open Access Journals (Sweden)

    Akram H. Aljahdali

    2017-04-01

    Full Text Available Closure of a giant omphalocele can be challenging. Preservation of the amnion in staged closure is not commonly practiced. Here, we describe 2 cases of giant omphalocele treated with a modified amnion preservation, staged closure technique. This paper demonstrates the feasibility and safety of this technique, and the versatility of amnion to adapt to an escharization strategy if closure is not achievable.

  2. Time-dependent mechanical behavior of human amnion: Macroscopic and microscopic characterization

    OpenAIRE

    2014-01-01

    © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved. Characterizing the mechanical response of the human amnion is essential to understand and to eventually prevent premature rupture of fetal membranes. In this study a large set of macroscopic and microscopic mechanical tests have been carried out on fresh unfixed amnion to gain insight into the time dependent material response and the underlying mechanisms. Creep and relaxation responses of amnion were characterized in...

  3. Gamma radiation sterilized amnion: use in ophthalmology

    Energy Technology Data Exchange (ETDEWEB)

    Martinez P, M. E. [ININ, Carretera Mexico-Toluca s/n, Ocoyoacac 52750, Estado de Mexico (Mexico); Leon T, Y. [Hospital General Regional 220, IMSS, Paseo Tollocan No. 620, Col. Vertice, Toluca 50150, Estado de Mexico (Mexico); Vazquez M, L., E-mail: esther.martinez@inin.gob.m [Hospital General de Mexico, Dr. Balmis 148, Col. Doctores, 06720 Mexico D. F. (Mexico)

    2010-10-15

    Amnion processed at the Radio sterilized Tissue Bank at the National Institute of Nuclear Research, sterilized with {sup 60}Co gamma radiation, have been used in Mexico since 2005 either as a graft to replace the damaged ocular surface, or as a patch to prevent unwanted inflammatory reactions. Patients from the Hospital General de Mexico (HGM) and Instituto Mexicano del Seguro Social (IMSS), suffering diverse pathologies such as keratoconjunctivitis; recurrent pterygium associated with symblepharon; corneal neuro trophic ulcers, chemical and thermal burns, and corneal thinning s, had been successfully treated with irradiated amnion. In the HGM, a clinical prospective study on lesions of the ocular surface of 17 eyes from 15 patients, affected with the above mentioned pathologies, was successful in 88.2%. The results have proven to be excellent as much for cosmetic purposes as for functional ones. Without the treatment, the patients could have suffered a healing after-effect or loss of sight. At IMSS, a controlled clinical randomized trial with 108 eyes from 100 patients, affected with primary nasal pterygium, was performed in 2009. These eyes were treated with radio sterilized amnion and intraoperative mitomycin C to prevent recurrence after excision of the primary pterygium. The preliminary results do not shown adverse reaction, inflammation and pain were significantly reduced radio sterilized amnion also offer security because they do no express antigens HLA-A, B or Dr and the sterile irradiated tissue do not provoke rejection or transmit an infective disease. (Author)

  4. Gamma radiation sterilized amnion: use in ophthalmology

    International Nuclear Information System (INIS)

    Martinez P, M. E.; Leon T, Y.; Vazquez M, L.

    2010-10-01

    Amnion processed at the Radio sterilized Tissue Bank at the National Institute of Nuclear Research, sterilized with 60 Co gamma radiation, have been used in Mexico since 2005 either as a graft to replace the damaged ocular surface, or as a patch to prevent unwanted inflammatory reactions. Patients from the Hospital General de Mexico (HGM) and Instituto Mexicano del Seguro Social (IMSS), suffering diverse pathologies such as keratoconjunctivitis; recurrent pterygium associated with symblepharon; corneal neuro trophic ulcers, chemical and thermal burns, and corneal thinning s, had been successfully treated with irradiated amnion. In the HGM, a clinical prospective study on lesions of the ocular surface of 17 eyes from 15 patients, affected with the above mentioned pathologies, was successful in 88.2%. The results have proven to be excellent as much for cosmetic purposes as for functional ones. Without the treatment, the patients could have suffered a healing after-effect or loss of sight. At IMSS, a controlled clinical randomized trial with 108 eyes from 100 patients, affected with primary nasal pterygium, was performed in 2009. These eyes were treated with radio sterilized amnion and intraoperative mitomycin C to prevent recurrence after excision of the primary pterygium. The preliminary results do not shown adverse reaction, inflammation and pain were significantly reduced radio sterilized amnion also offer security because they do no express antigens HLA-A, B or Dr and the sterile irradiated tissue do not provoke rejection or transmit an infective disease. (Author)

  5. Human amnion as a biological dressing used to prevent prolonged air leakage in thoracic surgery

    International Nuclear Information System (INIS)

    Mijewski, M.; Uhrynowska-Tyszkiewicz, I.; Piech, K.; Gogowski, M.

    2008-01-01

    Full text: Prolonged air leakage lasting 7 days or more is one of the most common complications in thoracic surgery. This complication may result in increased morbidity and prolonged hospital stay. Amnion allografts have been used to minimise this complication. The aim of our study was to evaluate the usefulness of human amnion grafts in the treatment of air leakage following thoracic surgery. Deep-frozen, radiation-sterilized (35 kGy) human amnion grafts prepared at the Central Tissue Bank in Warsaw (Poland) were used. Amnion allografts were applied to 69 patients who had surgery: 36 thoracotomies, and 33 rethoracotomies had been performed. During lung ventilation the air leakage sites were identified and covered by the amnion flap. Air leakage were evaluated during the postoperative period. Retrospectively we analysed air leakage duration in 170 thoracothomies and rethoracotomies without amnion transplantation. The separation of lung tissue and the liberation of pleural adhesions may be result in the lung and visceral pleura injury. Deep-frozen and radiation-sterilized human amnion is biocompatible, flexible, strong and airtight. It may be easily attached to the lung parenchyma and allows coverage of the area of the lung parenchyma deprived of the visceral pleura. The use of human amnion allografts is simple and safe. After treatment with amnion in 85% of the cases air leakage last less than 7 days, and only its traces were observed. Our results suggest that the human amnion grafts applied for the prevention of air leakage in lung surgery is a safe, simple and effective method. (Author)

  6. Amnion: a potent graft source for cell therapy in stroke.

    Science.gov (United States)

    Yu, Seong Jin; Soncini, Maddalena; Kaneko, Yuji; Hess, David C; Parolini, Ornella; Borlongan, Cesar V

    2009-01-01

    Regenerative medicine is a new field primarily based on the concept of transplanting exogenous or stimulating endogenous stem cells to generate biological substitutes and improve tissue functions. Recently, amnion-derived cells have been reported to have multipotent differentiation ability, and these cells have attracted attention as a novel cell source for cell transplantation therapy. Cells isolated from amniotic membrane can differentiate into all three germ layers, have low immunogenicity and anti-inflammatory function, and do not require the destruction of human embryos for their isolation, thus circumventing the ethical debate commonly associated with the use of human embryonic stem cells. Accumulating evidence now suggests that the amnion, which had been discarded after parturition, is a highly potent transplant material in the field of regenerative medicine. In this report, we review the current progress on the characterization of MSCs derived from the amnion as a remarkable transplantable cell population with therapeutic potential for multiple CNS disorders, especially stroke.

  7. Monovalent cations transfer through isolated human amnion: a new pharmacological model

    Energy Technology Data Exchange (ETDEWEB)

    Bara, M.; Guiet-Bara, A.; Durlach, J.

    1985-04-01

    Transfer of monovalent cations through the isolated human amnion consists of different factors: paracellular, coupling, ATPase dependent cellular transfer, leak cellular transfer. Understanding this transfer permits testing of the action of various substances. Physiological substances (Mg, taurine) increase ionic transfer and there is a vicarious effect between Mg and taurine. The tocolytic agents MgSO/sub 4/ and ethanol do not exhibit a good effect on the transfer: decrease with ethanol; equality between entry and exit fluxes with MgSO/sub 4/. On the other hand, amphotericin B increases mother-to-fetus transfer. Polluting metals (Pb, Cd, Hg, As) dramatically reduce exchanges and almost completely inhibit amnion permeability. Ingestion of ethanol also exhibits a dramatic effect on the exchange between mother and fetus through the amnion. Study of ionic transfer in vitro can be considered a pharmacological model to investigate the modifications of mother-fetus exchanges by various substances.

  8. Function and failure of the fetal membrane: Modelling the mechanics of the chorion and amnion.

    Directory of Open Access Journals (Sweden)

    Stefaan W Verbruggen

    Full Text Available The fetal membrane surrounds the fetus during pregnancy and is a thin tissue composed of two layers, the chorion and the amnion. While rupture of this membrane normally occurs at term, preterm rupture can result in increased risk of fetal mortality and morbidity, as well as danger of infection in the mother. Although structural changes have been observed in the membrane in such cases, the mechanical behaviour of the human fetal membrane in vivo remains poorly understood and is challenging to investigate experimentally. Therefore, the objective of this study was to develop simplified finite element models to investigate the mechanical behaviour and rupture of the fetal membrane, particularly its constituent layers, under various physiological conditions. It was found that modelling the chorion and amnion as a single layer predicts remarkably different behaviour compared with a more anatomically-accurate bilayer, significantly underestimating stress in the amnion and under-predicting the risk of membrane rupture. Additionally, reductions in chorion-amnion interface lubrication and chorion thickness (reported in cases of preterm rupture both resulted in increased membrane stress. Interestingly, the inclusion of a weak zone in the fetal membrane that has been observed to develop overlying the cervix would likely cause it to fail at term, during labour. Finally, these findings support the theory that the amnion is the dominant structural component of the fetal membrane and is required to maintain its integrity. The results provide a novel insight into the mechanical effect of structural changes in the chorion and amnion, in cases of both normal and preterm rupture.

  9. Clinical studies on using lyophilised radiation sterilised amnion membranes as dressing for leprosy wound

    International Nuclear Information System (INIS)

    Tarusarraya, P.; Basril, A.; Hilmy, N.

    1999-01-01

    Leprosy is a chronic disease caused by a bacillus Mycobacterium leprae and characterised by the formation of nodules or of macules that enlarge and spread accompanied by loss of sensation with eventual paralysis and production of deformities and mutilation. The wound of the disease can be treated by using conventional method such as Zinc Oxide (ZnO) Ointment with sterile gauze. Human amnion membranes have been used as a biological burn dressing with good results for several decades. A comparison study on using that conventional dressings and radiation sterilised lyophilised amnion membranes has been done at Sitanala Leprosarium to observe the effectiveness of using amnion membranes as leprosy wound dressing. Number of patients observed were 85, age from 12 to 60 years old. The locations of the wounds observed were at the leg and arm, with two types of wound i.e. reaction and simple ulcer. Parameter observed was the length of the healing time of the wounds. Results show that the average length of the healing time of the wound can be reduced from 64 days to 30 days when using amnion membranes compared to using the conventional wound dressing. The length of the healing time of the simple ulcer is longer that those of reaction wound using both of the dressings

  10. DREAM Is Involved in the Genesis of Inflammation-Induced Prolabour Mediators in Human Myometrial and Amnion Cells

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    Priyanka Goradia

    2018-01-01

    Full Text Available Preterm birth is the primary cause of perinatal morbidity and mortality worldwide. Inflammation induces a cascade of events leading to preterm birth by activating nuclear factor-κB (NF-κB. In nongestational tissues, downstream regulatory element antagonist modulator (DREAM regulates NF-κB activity. Our aims were to analyse DREAM expression in myometrium and fetal membranes obtained at term and preterm and to determine the effect of DREAM inhibition on prolabour mediators in primary myometrial and amnion cells. DREAM mRNA expression was significantly higher in fetal membranes obtained after spontaneous labour compared to nonlabour and in amnion from women with histological preterm chorioamnionitis when compared to amnion from women without chorioamnionitis. In primary myometrial and amnion cells, the effect of DREAM silencing by siRNA was a significant decrease in the expression of proinflammatory cytokine IL-6, the chemokines IL-8 and MCP-1, the adhesion molecule ICAM-1, MMP-9 mRNA expression and activity, and NF-κB transcriptional activity when stimulated with the proinflammatory cytokine IL-1β, the bacterial products fsl-1 or flagellin, or the viral dsRNA analogue poly(I:C. These data suggest that, in states of heightened inflammation, DREAM mRNA expression is increased and that, in myometrial and amnion cells, DREAM regulates proinflammatory and prolabour mediators which may be mediated via NF-κB.

  11. Use of gamma-irradiated amnion as a biological dressing in the management of radiation induced ulcers

    International Nuclear Information System (INIS)

    Astrid Lobo Gajiwala; Cynthia D Lima; Urmila Samant; Vilas Mayekar

    2008-01-01

    Full text: The treatment of moist skin desquamation (ulceration) following radiation therapy focuses on promoting healing, preventing microbial infections, protecting from mechanical trauma and providing pain relief. Various dressings like hydrocolloid, 1% gentian violet and other topical applications are routinely used. This study evaluated the use of freeze-dried, irradiated amnion from the Tata Memorial Hospital Tissue Bank in the management of radiation ulcers from 2001 to 2007. During this period, 2095 amnion dressings were used in 554 patients (238 male and 316 female). The ulcer was cleaned with normal saline and the amnion applied directly on the affected area. No topical antibiotics were used. The ulcer sites were in the regions of the groin and perianal area (40.79%), head and neck (30.50%), breast and axilla (18.59%), limbs (5.78%), chest (2.70%) and back (1.62%). Healing occurred within 4 days to 14 days using a single application of amnion in 291 patients (52.53%), 2 dressings in 129 patients (23.28%) and 3 dressings in 74 patients (13.36%). The remaining 60 patients (10.83%) required four or more dressings which were used over a period of a month. The amnion dressing was found to be convenient to apply and adhered well to the affected areas without the use of any adhesives unlike with routine hydro-colloid dressing. This was particularly advantageous in sites with irregular contours such as the groin and perianal areas and the head and neck regions (71.29%). Patients experienced almost immediate pain relief and the need for analgesics was reduced. In the majority of patients healing occurred earlier than was usually obtained with routine dressings. The amnion dressing proved to be advantageous over co ventional dressings because of its convenience of use and cost effectiveness due to shorter duration of healing, fewer dressing changes and diminished use of analgesics and antibiotics. (Author)

  12. Radiation sterilisation dose determination for lyophilised amnion membranes and lyophilised bone grafts

    International Nuclear Information System (INIS)

    Hilmy, N.; Basril, A.; Febrida, A.

    1999-01-01

    Radiation sterilisation of medical products is now well established in commercial scale and at present there are more than two hundred irradiation facilities in operation throughout the world. It is a cold sterilisation process without toxic chemical residues, high degree of safety and easy to control, so that it is a safe technology to sterilise human tissue grafts. According to ISO (International Organisation of Standard) No. 11137, radiation sterilisation dose should be established based on the number of product's bioburden (number of product's contaminated microbes before irradiation). Bioburden of lyophilised amnion membranes and lyophilised bone grafts produced by Batan Research Tissue Bank (BRTB) have been determined since 1990 and 1994 consecutively by using 100 up to 120 pieces of samples per year. Results show that the average bioburden of the amnion membranes were 1.4 ( 0.2 x 103; 1.2 (0.2 x 103; 1.2 ( 1.2 x 103; 4.5 ( 0.5 x 102; 1.8 ( 0.9 x 102; 2.4 ( 2.3 x 102; 1.7(l.5 x 102; 1.5 ( 1.7 x 102 cells per sample, calculated in 1990 to 1997 consecutively and the average bioburden of the bonegrafts were 1.5 (0.4x 101; 0.25 (0.12 x 101; 0; 0 cells per sample, calculated in 1994 to 1997 consecutively. Morphological of those contaminants were found to be Gram positive coccoid forms (98%) and Gram positive vegetative rod (2%) with the D10 - values of 0.25 to 0.50 kGy. No spore forming bacteria and Gram negative bacteria were found in those contaminations. The highest bioburden of lyophilised amnion membranes and lyophilised bone grafts were found to be 4900 and 80 cells per sample consecutively, and the lowest was found to be 0 cell per sample in both of materials observed. According to ISO 11137 radiation sterilisation doses for amnion membranes were ranging between 21 to 25 kGy and for bone grafts was around 15 kGy with the Sterility Assurance Level (SAL) of 106. Since 1990, radiation sterilisation dose used for lyophilised amnion membranes produced by BRTB

  13. The advantages of the application of amnion membrane in the treatment of burns.

    Science.gov (United States)

    Andonovska, D; Dzokic, Gj; Spasevska, L; Trajkovska, T; Popovska, K; Todorov, I; Petrovski, P; Kondov, G; Sapova, B; Marcikic, G; Atanasova, E; Obocki, E; Ugrinovska, J; Andonovski, D; Andonovski, D; Vasilevska, V; Mircevska-Zogovska, E

    2008-07-01

    A crucial and important factor for successful treatment of burns is the early covering of the burned area with skin substitutes. The covering of the burn requires material that restores the epidermal function and integrates itself into the process of healing. Biological dressings are the golden standard for the temporary covering of burns. All biological skin substitutes are susceptible to early graft reaction and the only exception is the amnion membrane. The importance of the amnion membrane as a biological dressing for burns amounts to: a barrier to bacterial colonization, hastens the epithelisation, and control of water loss. Amnioplasty is a method of application of amnion membrane on the recipient site. In this comparative study, 60 patients with dermal and sub-dermal burns were included. Research was made on an examination group of 30 patients with burns where the method of amnioplasty was applied, and for this amnion membrane conserved in 76% alcohol was used. The control group was made up of 30 patients with burns treated conventionally, and standard methods for the local treatment of burns were applied: exposition, occlusive dressing and initial excision with skin grafting. Pathohistological and microbiological analyses of the bioptical material were made. The degree of the burns was determined through a pathohistological analysis of the bioptical material taken the third day, and in some of the subjects where re-epithelialization was determined on the seventh day, the further re-epithelialization was observed clinically. Pathohistological examination enabled discrimination between bacterial colonization and the invasive bacterial infection. Furthermore, the type of bacterial colonization and infection was determined, which was confirmed with microbiological analysis. The analysis of the results from the microbiological and pathohistological researches of the bioptical material according to the bacterial colonization and infection showed that, although

  14. The growth of human fibroblasts and A431 epidermoid carcinoma cells on gamma-irradiated human amnion collagen substrata.

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    Liu, B; Harrell, R; Lamb, D J; Dresden, M H; Spira, M

    1989-10-15

    Human fibroblasts and A431 human epidermoid carcinoma cells were cultured on gamma-irradiated human amnion collagen as well as on plastic dishes and non-irradiated collagen coated dishes. The morphology, attachment, growth and short-term cytotoxicity of these culture conditions have been determined. Both irradiated and non-irradiated amnion collagen enhanced the attachment and proliferation of fibroblasts as compared to the plastic dishes. No differences in these properties were observed for A431 cells cultured on irradiated collagen when compared with culture on non-irradiated collagen substrates. Cytotoxicity assays showed that irradiated and non-irradiated collagens were not cytotoxic for either fibroblasts or A431 cells. The results demonstrated that amnion collagen irradiated at doses of 0.25-2.0 Mrads is optimal for cell growth.

  15. A comparative in vitro study of the viability of human keratinocytes grown on irradiated human amnion membrane and fibrin glue scaffolds

    International Nuclear Information System (INIS)

    Dorai, A.A.; Lim, C.K.; Azman, W.S.; Halim, A.S.

    2008-01-01

    Full text: The dried irradiated human amnion membrane has been used as a biological dressing for various clinical conditions. Being another biological membrane its potential as a scaffold to grow human keratinocytes is not known yet. To compare the growth patterns and cell viability of keratinocytes using fibrin glue and air dried amnion membrane as a scaffold. Keratinocytes were obtained from skin samples of six patients undergoing elective surgery. Fibrin glue (Tisseel, Baxter ) was diluted and used to coat the wells. Human dried amnion membrane was obtained and placed into the 24 well plates. Keratinocytes were seeded into the fibrin and amnion scaffold. Cell viability assay (MTT) was performed after 24, 48 and 72 hours. Finally the measurements were done by the Enzyme-Linked Immunosorbent Assay (ELISA) reader at 570 nm. Six patients consented for the study. The cells growing on the amnion scaffold showed a decreasing trend (20.67%, 17.94% and 16.78% respectively for 24, 48 and 72 hours). The cells growing on the fibrin scaffold showed a steady increase in number at 24, 48 and 72 hours (73.03%, 74.12% and 79.66%). The percentage of growth of normal human keratinocytes were significantly greater in the fibrin scaffold group (Mann - Whitney p = 0.002) for 24, 48 and 72 hours. The air dried irradiated human amnion membrane can be used as a scaffold to grow keratinocytes but however the growth pattern does not sustain with time. Fibrin glue supports the growth of human keratinocytes and shows an increasing pattern of growth with time. (Author)

  16. Timing of Histologic Progression from Chorio-Deciduitis to Chorio-Deciduo-Amnionitis in the Setting of Preterm Labor and Preterm Premature Rupture of Membranes with Sterile Amniotic Fluid.

    Science.gov (United States)

    Park, Chan-Wook; Park, Joong Shin; Norwitz, Errol R; Moon, Kyung Chul; Jun, Jong Kwan; Yoon, Bo Hyun

    2015-01-01

    Histologic chorio-deciduitis and chorio-deciduo-amnionitis (amnionitis) in extra-placental membranes are known to represent the early and advanced stages of ascending intra-uterine infection. However, there are no data in humans about the time required for chorio-deciduitis to develop and for chorio-deciduitis without amnionitis to progress to chorio-deciduitis with amnionitis, and the effect of prolongation of pregnancy on the development of chorio-deciduitis and amnionitis in patients with preterm labor and intact membranes (PTL) and preterm premature rupture of membranes (preterm-PROM). We examined these issues in this study. The study population consisted of 289 women who delivered preterm (133 cases with PTL, and 156 cases with preterm-PROM) and who had sterile amniotic fluid (AF) defined as a negative AF culture and the absence of inflammation as evidenced by a matrix metalloproteinase-8 (MMP-8) level membranes (i.e., inflammation-free extra-placental membranes, choroi-deciduitis only, and chorio-deciduitis with amnionitis) in patients with PTL and preterm-PROM. Amniocentesis-to-delivery interval was longer in cases of chorio-deciduitis with amnionitis than in cases of chorio-deciduitis only in both PTL (median [interquartile-range (IQR)]; 645.4 [319.5] vs. 113.9 [526.9] hours; P = 0.005) and preterm-PROM (131.3 [135.4] vs. 95.2 [140.5] hours; Pmembranes. Moreover, prolongation of pregnancy is an independent predictor of the development of both chorio-deciduitis and amnionitis in cases of PTL with sterile AF.

  17. Evaluation of the cranial base in amnion rupture sequence involving the anterior neural tube: implications regarding recurrence risk.

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    Jones, Kenneth Lyons; Robinson, Luther K; Benirschke, Kurt

    2006-09-01

    Amniotic bands can cause disruption of the cranial end of the developing fetus, leading in some cases to a neural tube closure defect. Although recurrence for unaffected parents of an affected child with a defect in which the neural tube closed normally but was subsequently disrupted by amniotic bands is negligible; for a primary defect in closure of the neural tube to which amnion has subsequently adhered, recurrence risk is 1.7%. In that primary defects of neural tube closure are characterized by typical abnormalities of the base of the skull, evaluation of the cranial base in such fetuses provides an approach for making a distinction between these 2 mechanisms. This distinction has implications regarding recurrence risk. The skull base of 2 fetuses with amnion rupture sequence involving the cranial end of the neural tube were compared to that of 1 fetus with anencephaly as well as that of a structurally normal fetus. The skulls were cleaned, fixed in 10% formalin, recleaned, and then exposed to 10% KOH solution. After washing and recleaning, the skulls were exposed to hydrogen peroxide for bleaching and photography. Despite involvement of the anterior neural tube in both fetuses with amnion rupture sequence, in Case 3 the cranial base was normal while in Case 4 the cranial base was similar to that seen in anencephaly. This technique provides a method for determining the developmental pathogenesis of anterior neural tube defects in cases of amnion rupture sequence. As such, it provides information that can be used to counsel parents of affected children with respect to recurrence risk.

  18. A roentgenographic assessment of regenerative efficacy of bioactive Gengigel® in conjunction with amnion membrane in grade II furcation defect

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    S Harveen Kalra

    2015-01-01

    Full Text Available Background: Nowadays, techniques are being developed to guide and instruct the specialized cellular components of the periodontium to participate in the regenerative process. This approach of reconstruction makes use of understanding of the development of the periodontium and the cellular processes that are involved. Hyaluronic acid is a naturally occurring non-sulfated high molecular weight glycosaminoglycan that forms a critical component of the extracellular matrix and contributes significantly to tissue hydrodynamics, cell migration, and proliferation. Hence, its administration to periodontal wound sites could achieve comparable beneficial effects in periodontal tissue regeneration. Hence, the purpose of the present case report was to assess roentgenographically, the regenerative capacity of Gengigel® in conjunction with bioactive amnion guided tissue regeneration (GTR membrane in a patient with Grade II furcation defect. Case Presentation: A patient complained of bleeding gums from the lower back tooth region, reportedly found Grade II furcation in the lower right mandibular first molar. After Phase, I therapy, Gengigel® along with bioactive amnion membrane was placed in the furcation area during the surgical phase. Roentgenographic assessment was done at 4 months and 6 months postoperatively. It resulted in complete defect-fill and loss of radiolucency at 6 months. Conclusion: Surgical placement of Gengigel® along with amnion membrane in the furcation defect can significantly improve the periodontal defect morphology.

  19. Amnion as a surrogate tissue reporter of the effects of maternal preeclampsia on the fetus.

    Science.gov (United States)

    Suzuki, Masako; Maekawa, Ryo; Patterson, Nicole E; Reynolds, David M; Calder, Brent R; Reznik, Sandra E; Heo, Hye J; Einstein, Francine Hughes; Greally, John M

    2016-01-01

    Preeclampsia, traditionally characterized by high blood pressure and proteinuria, is a common pregnancy complication, which affects 2-8 % of all pregnancies. Although children born to women with preeclampsia have a higher risk of hypertension in later life, the mechanism of this increased risk is unknown. DNA methylation is an epigenetic modification that has been studied as a mediator of cellular memory of adverse exposures in utero. Since each cell type in the body has a unique DNA profile, cell subtype composition is a major confounding factor in studies of tissues with heterogeneous cell types. The best way to avoid this confounding effect is by using purified cell types. However, using purified cell types in large cohort translational studies is difficult. The amnion, the inner layer of the fetal membranes of the placenta, is derived from the epiblast and consists of two cell types, which are easy to isolate from the delivered placenta. In this study, we demonstrate the value of using amnion samples for DNA methylation studies, revealing distinctive patterns between fetuses exposed to proteinuria or hypertension and fetuses from normal pregnancies. We performed a genome-wide DNA methylation analysis, HpaII tiny fragment Enrichment by Ligation-mediated PCR (HELP)-tagging, on 62 amnion samples from the placentas of uncomplicated, normal pregnancies and from those with complications of preeclampsia or hypertension. Using a regression model approach, we found 123, 85, and 99 loci with high-confidence hypertension-associated, proteinuria-associated, and hypertension- and proteinuria-associated DNA methylation changes, respectively. A gene ontology analysis showed DNA methylation changes to be selecting genes with different biological processes in exposure status. We also found that these differentially methylated regions overlap loci previously reported as differentially methylated regions in preeclampsia. Our findings support prior observations that preeclampsia

  20. Regenerative and Antibacterial Properties of Acellular Fish Skin Grafts and Human Amnion/Chorion Membrane: Implications for Tissue Preservation in Combat Casualty Care.

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    Magnusson, Skuli; Baldursson, Baldur Tumi; Kjartansson, Hilmar; Rolfsson, Ottar; Sigurjonsson, Gudmundur Fertram

    2017-03-01

    Improvised explosive devices and new directed energy weapons are changing warfare injuries from penetrating wounds to large surface area thermal and blast injuries. Acellular fish skin is used for tissue repair and during manufacturing subjected to gentle processing compared to biologic materials derived from mammals. This is due to the absence of viral and prion disease transmission risk, preserving natural structure and composition of the fish skin graft. The aim of this study was to assess properties of acellular fish skin relevant for severe battlefield injuries and to compare those properties with those of dehydrated human amnion/chorion membrane. We evaluated cell ingrowth capabilities of the biological materials with microscopy techniques. Bacterial barrier properties were tested with a 2-chamber model. The microstructure of the acellular fish skin is highly porous, whereas the microstructure of dehydrated human amnion/chorion membrane is mostly nonporous. The fish skin grafts show superior ability to support 3-dimensional ingrowth of cells compared to dehydrated human amnion/chorion membrane (p fish skin is a bacterial barrier for 24 to 48 hours. The unique biomechanical properties of the acellular fish skin graft make it ideal to be used as a conformal cover for severe trauma and burn wounds in the battlefield. Reprint & Copyright © 2017 Association of Military Surgeons of the U.S.

  1. Amnion and Chorion Membranes: Potential Stem Cell Reservoir with Wide Applications in Periodontics.

    Science.gov (United States)

    Gupta, Akanksha; Kedige, Suresh D; Jain, Kanu

    2015-01-01

    The periodontal therapy usually aims at elimination of disease causing bacteria and resolution of inflammation. It involves either resective or regenerative surgery to resolve the inflammation associated defects. Over the years, several methods have been used for achievement of periodontal regeneration. One of the oldest biomaterials used for scaffolds is the fetal membrane. The amniotic membranes of developing embryo, that is, amnion (innermost lining) and chorion (a layer next to it), have the properties with significant potential uses in dentistry. This paper reviews the properties, mechanism of action, and various applications of these placental membranes in general and specifically in Periodontics.

  2. Human amnion epithelial cells expressing HLA-G as novel cell-based treatment for liver disease.

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    Strom, Stephen C; Gramignoli, Roberto

    2016-09-01

    Despite routine liver transplantation and supporting medical therapies, thousands of patients currently wait for an organ and there is an unmet need for more refined and widely available regenerative strategies to treat liver diseases. Cell transplants attempt to maximize the potential for repair and/or regeneration in liver and other organs. Over 40years of laboratory pre-clinical research and 25years of clinical procedures have shown that certain liver diseases can be treated by the infusion of isolated cells (hepatocyte transplant). However, like organ transplants, hepatocyte transplant suffers from a paucity of tissues useful for cell production. Alternative sources have been investigated, yet with limited success. The tumorigenic potential of pluripotent stem cells together with their primitive level of hepatic differentiation, have limited the use of stem cell populations. Stem cell sources from human placenta, and the amnion tissue in particular are receiving renewed interest in the field of regenerative medicine. Unlike pluripotent stem cells, human amnion epithelial (AE) cells are easily available without ethical or religious concerns; they do not express telomerase and are not immortal or tumorigenic when transplanted. In addition, AE cells have been reported to express genes normally expressed in mature liver, when transplanted into the liver. Moreover, because of the possibility of an immune-privileged status related to their expression of HLA-G, it might be possible to transplant human AE cells without immunosuppression of the recipient. Copyright © 2016. Published by Elsevier Inc.

  3. Influence of physiologically active complex isolated from human amnion on lipid peroxide oxidation state and antioxidant activity of blood in rats after irradiation in different doses

    International Nuclear Information System (INIS)

    Borshchevs'ka, M.Yi.; Popov, V.V.; Abramova, L.P.; Kuz'myinova, Yi.A.

    1995-01-01

    The authors have studied the influence of physiologically active complex isolated from human amnion on the state of lipid peroxide oxidation according to diene conjugate and malonic dialdehyde amount and antioxidant enzyme activity (catalase and glutationperoxidase) in the blood of the rats exposed to single total irradiation in different doses (4 and 6 Gy) was studied. Definite changes of peroxide process intensity and reduction of the enzymes activity were shown to be observed in the blood of experimental animals even at long terms after the radiation exposure. Under the background of radiation exposure, administration of physiologically active complex isolated from human amnion produced protective effect on antioxidant enzyme activity which promoted normalization of peroxidation processes within the post-radiation period

  4. Amnioinfusion in preterm PROM: effects on amnion and cord histology.

    Science.gov (United States)

    Locatelli, A; Andreani, M; Ghidini, A; Verderio, M; Pizzardi, A; Vergani, P; Salafia, C M

    2008-02-01

    To investigate the effects of transabdominal amnioinfusion (TA) on the histology of amnion (A) and umbilical cord (UC). From a cohort of 56 singleton pregnancies with premature rupture of membranes (PROM) at

  5. Pregnancy outcome of monochorionic twins: does amnionicity matter?

    Science.gov (United States)

    Dias, Thiran; Contro, Elena; Thilaganathan, Basky; Khan, Hina; Zanardini, Cristina; Mahsud-Dornan, Samina; Bhide, Amar

    2011-12-01

    To compare the fetal loss rate of monochorionic (MC) twin pregnancies according to their amnionicity. A retrospective review of all MC pregnancy outcomes in a tertiary centre. Pregnancy outcomes were compared for monochorionic monoamniotic (MCMA) versus monochorionic diamniotic (MCDA) pregnancies. 29 MCMA and 117 MCDA twin pregnancies were identified. The overall fetal loss rate was significantly higher in MCMA (23/52, 44.2%) compared to MCDA pregnancies (28/233, 12%, Chi squared = 30.03, p fetal survival rate in MCDA twins were significantly higher than in MCMA twins (Log-rank Chi-squared = 27.9, p fetal losses in some MCMA twins. After exclusion of identifiable causes, the difference in fetal survival was not significant in the two groups (Log-rank chi-squared = 0.373, p = .54). The loss rate for MCMA twins is high and occurs mainly due to discordant congenital abnormality, conjoint twins or twin reversed arterial perfusion (TRAP) sequence. Although the fetal loss rate in MCDA is lower than in MCMA pregnancies, the majority of fetal loss in MCDA pregnancies cannot be predicted at the first scan at presentation. The data of this study questions the widespread policy of a difference in the scheduling of elective delivery for MCMA and MCDA twins.

  6. Human amnion mesenchymal stem cells promote proliferation and osteogenic differentiation in human bone marrow mesenchymal stem cells.

    Science.gov (United States)

    Wang, Yuli; Yin, Ying; Jiang, Fei; Chen, Ning

    2015-02-01

    Human amnion mesenchymal stem cells (HAMSCs) can be obtained from human amniotic membrane, a highly abundant and readily available tissue. HAMSC sources present fewer ethical issues, have low immunogenicity, anti-inflammatory properties, considerable advantageous characteristics, and are considered an attractive potential treatment material in the field of regenerative medicine. We used a co-culture system to determine whether HAMSCs could promote osteogenesis in human bone marrow mesenchymal stem cells (HBMSCs). We isolated HAMSCs from discarded amnion samples and collected them using pancreatin/collagenase digestion. We cultured HAMSCs and HBMSCSs in basal medium. Activity of alkaline phosphatase (ALP), an early osteogenesis marker, was increased in the co-culture system compared to the control single cultures, which we also confirmed by ALP staining. We used immunofluorescence testing to investigate the effects of co-culturing with HAMSCs on HBMSC proliferation, which revealed that the co-culturing enhanced EdU expression in HBMSCs. Western blotting and quantitative real-time PCR indicated that co-culturing promoted osteogenesis in HBMSCs. Furthermore, Alizarin red S staining revealed that extracellular matrix calcium levels in mineralized nodule formation produced by the co-cultures were higher than that in the controls. Using the same co-culture system, we further observed the effects of HAMSCs on osteogenic differentiation in primary osteoblasts by Western blotting, which better addressed the mechanism for HAMSCs in bone regeneration. The results showed HAMSCs are osteogenic and not only play a role in promoting HBMSC proliferation and osteogenic differentiation but also in osteoblasts, laying the foundation for new regenerative medicine methods.

  7. The correlation between histologic placentitis and amnionitis and the amnioniotic fluid's inflammatory cytokines in case of spontaneous pre-term labor with intact membrane

    Directory of Open Access Journals (Sweden)

    Agus Abadi

    2001-12-01

    Full Text Available Pre-term labor is presumed to result from spreading of lower genital infection to upper part, subsequently to decidual and choioamniotic tissues. Host response to this injury include the expression of protein which is responsible to the inflammatory reactions. The expression of the inflammatory cytokines such as IL-1β, IL-6, IL-8 and TNF-α increase in case of infection.These cytokines may play an essential role in the pathophysiology of spontaneous pretem labor with intact membrane.An observational analytic cohort study was caried out on cases of spontaneous pre-tefln labor with intact membrane. The objectives of this study are to examine the relationship between l the histologic amnionitis and placentitis and the incidence of preterm delivery,2 the expression of amniotic fluid's IL-1β, IL-6, IL-8 and TNF-α and the incidence of preterm delivery, 3 the level of amniotic fluid's IL-1β, IL-6, IL-8 and TNF-α and the grade of histologic amnionitis and placentitis in case of pre-term labor with intact membrane. Cases of spontaneous Pre'teftn labor with intact membrane which underwent transabdominal amniocentesis at admission and managed as standard procedure for pre-term labor with intact membrane. Atl of the cases were observed until the delivery of the baby, eithir preterm or term. The membrane and the placentawere cut postnatally and then the histologic acute inflammation eyaluated based on the criteria of Salafia.The level of amniotic fluid IL-1β, IL-6, IL-8 and TNF-α were analyzed quantitatively by Elisa method. This study showed thet the degree of histologic amnionitis and placentitis, and the level of amniotic fluid's IL-1β, IL-6, IL-8 and TNF-α were significantly higher in pre-term compared to terrn deliveries (p<0.05 and lhere were a positive correlation between the grade of histoLogic inflammation and the level of amniotic fluid's cytokines (Spearmann Rank Conelation test; p<0,05 in cases of preterm labor with intact membrane. The

  8. Syndecan expressions in the human amnion and chorionic plate

    Directory of Open Access Journals (Sweden)

    T. Lorenzi

    2010-10-01

    Full Text Available The syndecan family consists of four distinct membrane glycoproteins in mammals. Syndecans control cell proliferation, differentiation, adhesion and migration through participation in cell-cell interactions, anchorage of cells to the extracellular environment, and modulation of multiple growth factors. Therefore, syndecans may play a pivotal role in the regulation of cell behaviour depending on the cellular microenvironment. Here, we demonstrate that syndecan-1, syndecan-2 and syndecan-4 are expressed in fetal membrane tissue with different immunolocalizations. Syndecan-1 is expressed in the amniotic epithelium, localizing at basolateral cell surfaces. Syndecan-2 and syndecan-4, in contrast, are mostly localized in intracellular compartments, in the extravillous cytotrophoblastic cells and in some fibroblasts of the chorionic plate as well as in the amniotic epithelial cells. In the latter, syndecan-4 is mainly localized in the apical part of the cells. Our results strongly suggest a key role of syndecan-1, syndecan-2 and syndecan-4 in the determination of structural and functional characteristics of human amnion and chorionic plate. Since the solute exchanges between fetus and mother take place in fetal membranes, our data suggest that syndecans are important players in the placenta for the establishment of the fetal-maternal inter-communication.

  9. Brief communication: sliding displacement of amnion and chorion following controlled laser wounding suggests a mechanism for short-term sealing of ruptured membranes.

    Science.gov (United States)

    Behzad, F; Dickinson, M R; Charlton, A; Aplin, J D

    1994-10-01

    The Erbium-YAG laser was used to produce narrow wounds of defined depth in term amniochorion. The charring effect of the laser meant that sites could be readily localized in histological sections. During brief post-wounding incubations, sliding displacement of the amnion relative to the chorion occurred through the plane of the spongy layer. This suggests a possible short-term mechanism whereby a spontaneous rupture could be sealed in vivo.

  10. Amnion s and radio-sterilized porcine skin use as potential matrices for the development of human skin substitutes

    International Nuclear Information System (INIS)

    Martinez P, M. E.; Reyes F, M. L.; Reboyo B, D.; Velasquillo M, M. C.; Sanchez S, R.; Brena M, A. M.; Ibarra P, J. C.

    2014-10-01

    The injuries by burns constitute a primordial problem of public health; they cause a high mortality index, severe physical and psychological disability, etc. The autologous skin transplant is the replacement therapy recommended for its treatment, but in patients that present a high percentage of burnt skin; this is not possible to carry out. Another strategy is the transplant of donated skin; however, due to the little donation that exists in our country is not very feasible to apply this treatment. A challenge of the tissues engineering is to develop biological skin substitutes, based on cells and amnion s, favoring the cutaneous regeneration and quick repair of injuries, diminishing this way the hospitalization expenses. At present skin substitutes that can equal to the same skin do not exist. On the other hand, the mesenchymal stromal cells (Msc) represent an alternative to achieve this objective; since has been demonstrated that the Msc participate in the tissue repair by means of inhibition of pro-inflammatory cytokines and differentiation to dermal fibroblasts and keratinocytes. To apply the Msc in cutaneous injuries a support material is required that to allow transplanting these cells to a lesion or burn. The radio-sterilized human amnion and the radio-sterilized porcine skin, processed by the Radio-Sterilized Tissues Bank of the Instituto Nacional de Investigaciones Nucleares (ININ), are biomaterials that are used as temporary cutaneous coverings. We suppose that these two matrices will be appropriate for the growth and maintenance in cultivation of the Msc, to generate two biological skin substitutes, in collaboration with the Biotechnology Laboratory of the Instituto Nacional de Rehabilitacion. (Author)

  11. Comparison of the effects of taurine and magnesium on electrical characteristics of artificial and natural membranes. V. Study on the human amnion of the antagonism between magnesium, taurine and polluting metals

    Energy Technology Data Exchange (ETDEWEB)

    Bara, M.; Guiet-Bara, A.; Durlach, J.

    1985-01-01

    The effects of metal pollutants (Pb, Cd, Hg, As) were studied on strips of human amnion isolated from the placental zone put in between two Ussing chambers with Hanks' solution at 37 degrees C and pH 7.4. The total conductance Gt through the human isolated amnion was decreased on the fetal side by Pb and As; on the maternal side by Cd, Hg and As. When Gt was decreased by metal pollutants, Mg or taurine (TA) were added in the external medium to induce an antagonism between Mg or TA and metal pollutants. The addition of Mg increased significantly the Gt reduced by Pb, Cd and Hg, but had no effect on the Gt reduced by As. The addition of taurine increased significantly the Gt reduced by Cd and Hg, but had no effect on the Gt reduced by Pb and As. Dixon's kinetics (Gt as a function of the Mg or TA concentration when the metal pollutant concentration increased) indicate that there is a competitive inhibition between Mg-Pb and Mg-Cd (the inhibition constant Ki is lower with Pb (= 2.5) than with Cd (= 11.4) and suggests a greater antagonism between Mg-Pb than between Mg-Cd). Moreover, there appears to be a noncompetitive inhibition between Mg-Hg, TA-Cd and TA-Hg. These results indicate that Mg and TA, on the fetal side, exert an action on the same sites and that, on the maternal side, their action takes place on the same sites and also on different ones. Also, TA can be considered as a partial magnesium agonist, at least in the human amnion.

  12. Retinoid inhibition of in vitro invasion of human amnion basement membrane by human tumor cells

    International Nuclear Information System (INIS)

    Fazely, F.; Ledinko, N.; Smith, D.J.

    1986-01-01

    The biological activity of retinoids was assayed in an in vitro quantitative assay of human tumor cell invasion using human amnion basement membrane (BM). The effects measured were the inhibition of tumor cell migration through the BM and tumor cell degradative enzyme activity on 14 C-proline labeled collagenous and noncollagenous components of the BM. The human lung carcinoma A549 or the human Ewing's sarcoma TC-106 cell lines treated with retinoids for two days were incubated on the BM in the absence of retinoids. A dose-dependent inhibition of cell invasion was produced by retinoids. Among the retinoids tested, the most powerful was retinol acetate which inhibited invasion by 50% of A549 cells at a concentration of 0.009 μg/mL, and of TC-106 cells at 0.07 μg/mL. Retinol acetate inhibited A549 and TC-106 cell growth by approximately 50% at levels over 100-fold higher than those needed for antiinvasive activity. Retinol acetate was about 20 times more potent than retinoic acid and 30 times more potent than retinol palmitate. The model system will be useful for investigating antiinvasive activity of other retinoids as well as other compounds

  13. Effect of barbiturates on radiosensitivity of cells: a comparative study of electrophoretic mobility, colony forming ability and thymidine uptake on human amnion cells

    International Nuclear Information System (INIS)

    Lalwani, N.D.; Chaubal, K.A.

    1980-01-01

    Suspensions of human amnion cells were 60 Co γ-irradiated in the presence of phenobarbital or thiobarbital (50 μg/ml). The barbiturates protected the cells against the dose-dependent reduction in electrophoretic mobility (EPM) observed 4 hours after irradiation of untreated cells, although there was an initial decrease in the EPM of treated cells followed by recovery. Treated irradiated cells exhibited greater colony-forming ability than the untreated cells. Pentobarbital and phenobarbital had similar effects, but thiobarbital was not so effective. 3 H-TdR uptake increased within 4 hours of irradiation for the treated cells. The reproductive capacity of the cells was retained at doses as high as 500 rad. The results are discussed with reference to the effects of anaesthetics on cell membranes. (U.K.)

  14. Enantioselective cytotoxicity of the insecticide bifenthrin on a human amnion epithelial (FL) cell line

    International Nuclear Information System (INIS)

    Liu Huigang; Zhao Meirong; Zhang Cong; Ma Yun; Liu Weiping

    2008-01-01

    Synthetic pyrethroids (SPs) are used in preference to organochlorines and organophosphates due to their high efficiency, low toxicity to mammals, and ready biodegradability. Previous studies reported that enantioselective toxicity of SPs occurs in aquatic toxicity. Several studies have indicated that SPs could lead to oxidative damage in humans or animals which was associated with their toxic effects. Little is known about the differences in the effects of chronic toxicity induced by individual stereoisomers of chiral SPs. The present study was therefore undertaken to evaluate the enantioselectivity in cytotoxicity, genotoxicity caused by bifenthrin (BF) on human amnion epithelial (FL) cell lines and pesticidal activity on target organism. The cell proliferation and cytoflow analysis indicated that 1S-cis-BF presented more toxic effects than 1R-cis-BF above the concentration of 7.5 mg L -1 (p > 0.05). FL cells incubated with 1S-cis-BF exhibited a dose-dependent accumulation of intracellular reactive oxygen species (ROS). In the comet assay, the number of cells with damaged DNA incubated with 1S-cis-BF was more than that with 1R-cis-BF (p 50 values of enantiomer to the target pest on Pieris rapae L. show that 1R-cis-BF was 300 times more active than 1S-cis-BF. These results indicate that the enantioselective toxicity and activity of BF between non-target organism and target organism was reversal. These implications together suggest that assessment of the environmental safety and new pesticides development with chiral centers should consider enantioselectivity

  15. Experiment of amnion epithelial cell suspension liquid used for acute rabbit corneal alkali burn

    Directory of Open Access Journals (Sweden)

    Yan-Yan Zhang

    2017-10-01

    Full Text Available AIM: To investigate the effects of amnion epithelial cell(AECsuspension liquid on the biological behavior of the rabbit's corneal epithelium, combined with the in vitro and in vivo experiments. METHODS: The rabbit's corneal epithelium were cultured in the lower chamber of transwell, and AEC suspension liquid was dropwised in the upper chamber. There was only culture medium in the upper chamber of the control group. The proliferation of rabbit's corneal epithelium was observed with CCK-8 automated colorimetry and the expression of PCNA was detected by immunocytochemistry. We used the scratch wound assay to detect the migration of corneal epithelial cell(CEC. The in vivo models were established by placing a 10mm diameter corneal trephine in the center of the cornea, within 1mol/L NaOH for 1min. We divided those into three groups: treatment group of AEC suspension liquid eye drop, AEC suspension liquid subconjunctival injection and the control group without any treatment. Using the slit-lamp biomicroscope and fluorescence staining to observe the cornea per week. After 28d we took the eyeballs with the HE staining. The expression of VEGF was detected by immunohistochemistry. RESULTS: The activity of CEC with AEC treatment was much higher than the control group(PPIn vivo, the inflammation of the corneal and the CNV of the AEC group were all significantly reduced compared with the control group(PPCONCLUSION: AEC suspension liquid can promote the proliferation and migration of the rabbit's corneal epithelium. The potential of AEC suspension liquid as a therapy for acute corneal alkali burn.

  16. Contribution of aquaporin 9 and multidrug resistance-associated protein 2 to differential sensitivity to arsenite between primary cultured chorion and amnion cells prepared from human fetal membranes

    International Nuclear Information System (INIS)

    Yoshino, Yuta; Yuan, Bo; Kaise, Toshikazu; Takeichi, Makoto; Tanaka, Sachiko; Hirano, Toshihiko; Kroetz, Deanna L.; Toyoda, Hiroo

    2011-01-01

    Arsenic trioxide (arsenite, As III ) has shown a remarkable clinical efficacy, whereas its side effects are still a serious concern. Therefore, it is critical to understand the effects of As III on human-derived normal cells for revealing the mechanisms underlying these side effects. We examined the effects of As III on primary cultured chorion (C) and amnion (A) cells prepared from human fetal membranes. A significant dose-dependent As III -mediated cytotoxicity was observed in the C-cells accompanied with an increase of lactate dehydrogenase (LDH) release. Higher concentrations of As III were required for the A-cells to show cytotoxicity and LDH release, suggesting that the C-cells were more sensitive to As III than the A-cells. The expression levels of aquaporin 9 (AQP9) were approximately 2 times higher in the C-cells than those in the A-cells. Both intracellular arsenic accumulation and its cytotoxicity in the C-cells were significantly abrogated by sorbitol, a competitive AQP9 inhibitor, in a dose-dependent manner. The protein expression levels of multidrug resistance-associated protein (MRP) 2 were downregulated by As III in the C-cells, but not in the A-cells. No significant differences in the expression levels of MRP1 were observed between C- and A-cells. The protein expression of P-glycoprotein (P-gp) was hardly detected in both cells, although a detectable amount of its mRNA was observed. Cyclosporine A, a broad-spectrum inhibitor for ABC transporters, and MK571, a MRP inhibitor, but not PGP-4008, a P-gp specific inhibitor, potently sensitized both cells to As III -mediated cytotoxicity. These results suggest that AQP9 and MRP2 are involved in controlling arsenic accumulation in these normal cells, which then contribute to differential sensitivity to As III cytotoxicity between these cells. -- Highlights: ► Examination of effect of As III on primary cultured chorion (C) and amnion (A) cells. ► Dose-dependent As III -mediated cytotoxicity in C

  17. Amnion s and radio-sterilized porcine skin use as potential matrices for the development of human skin substitutes; Uso de amnios y piel porcina radioesterilizados como matrices potenciales para el desarrollo de sustitutos de piel humana

    Energy Technology Data Exchange (ETDEWEB)

    Martinez P, M. E.; Reyes F, M. L.; Reboyo B, D. [ININ, Carretera Mexico-Toluca s/n, 52750 Ocoyoacac, Estado de Mexico (Mexico); Velasquillo M, M. C.; Sanchez S, R.; Brena M, A. M.; Ibarra P, J. C., E-mail: esther.martinez@inin.gob.mx [Instituto Nacional de Rehabilitacion, Calz. Mexico-Xochimilco No. 289, Col. Arenal de Guadalupe, 14389 Mexico D. F. (Mexico)

    2014-10-15

    The injuries by burns constitute a primordial problem of public health; they cause a high mortality index, severe physical and psychological disability, etc. The autologous skin transplant is the replacement therapy recommended for its treatment, but in patients that present a high percentage of burnt skin; this is not possible to carry out. Another strategy is the transplant of donated skin; however, due to the little donation that exists in our country is not very feasible to apply this treatment. A challenge of the tissues engineering is to develop biological skin substitutes, based on cells and amnion s, favoring the cutaneous regeneration and quick repair of injuries, diminishing this way the hospitalization expenses. At present skin substitutes that can equal to the same skin do not exist. On the other hand, the mesenchymal stromal cells (Msc) represent an alternative to achieve this objective; since has been demonstrated that the Msc participate in the tissue repair by means of inhibition of pro-inflammatory cytokines and differentiation to dermal fibroblasts and keratinocytes. To apply the Msc in cutaneous injuries a support material is required that to allow transplanting these cells to a lesion or burn. The radio-sterilized human amnion and the radio-sterilized porcine skin, processed by the Radio-Sterilized Tissues Bank of the Instituto Nacional de Investigaciones Nucleares (ININ), are biomaterials that are used as temporary cutaneous coverings. We suppose that these two matrices will be appropriate for the growth and maintenance in cultivation of the Msc, to generate two biological skin substitutes, in collaboration with the Biotechnology Laboratory of the Instituto Nacional de Rehabilitacion. (Author)

  18. Inflammatory gene regulatory networks in amnion cells following cytokine stimulation: translational systems approach to modeling human parturition.

    Directory of Open Access Journals (Sweden)

    Ruth Li

    Full Text Available A majority of the studies examining the molecular regulation of human labor have been conducted using single gene approaches. While the technology to produce multi-dimensional datasets is readily available, the means for facile analysis of such data are limited. The objective of this study was to develop a systems approach to infer regulatory mechanisms governing global gene expression in cytokine-challenged cells in vitro, and to apply these methods to predict gene regulatory networks (GRNs in intrauterine tissues during term parturition. To this end, microarray analysis was applied to human amnion mesenchymal cells (AMCs stimulated with interleukin-1β, and differentially expressed transcripts were subjected to hierarchical clustering, temporal expression profiling, and motif enrichment analysis, from which a GRN was constructed. These methods were then applied to fetal membrane specimens collected in the absence or presence of spontaneous term labor. Analysis of cytokine-responsive genes in AMCs revealed a sterile immune response signature, with promoters enriched in response elements for several inflammation-associated transcription factors. In comparison to the fetal membrane dataset, there were 34 genes commonly upregulated, many of which were part of an acute inflammation gene expression signature. Binding motifs for nuclear factor-κB were prominent in the gene interaction and regulatory networks for both datasets; however, we found little evidence to support the utilization of pathogen-associated molecular pattern (PAMP signaling. The tissue specimens were also enriched for transcripts governed by hypoxia-inducible factor. The approach presented here provides an uncomplicated means to infer global relationships among gene clusters involved in cellular responses to labor-associated signals.

  19. Contribution of aquaporin 9 and multidrug resistance-associated protein 2 to differential sensitivity to arsenite between primary cultured chorion and amnion cells prepared from human fetal membranes

    Energy Technology Data Exchange (ETDEWEB)

    Yoshino, Yuta [Department of Clinical Molecular Genetics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan); Yuan, Bo, E-mail: yuanbo@toyaku.ac.jp [Department of Clinical Molecular Genetics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan); Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, 1550 4th St, RH584E Box 2911 San Francisco, CA 94158-2911 (United States); Kaise, Toshikazu [Laboratory of Environmental Chemodynamics, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan); Takeichi, Makoto [Yoneyama Maternity Hospital, 2-12 Shin-machi, Hachioji, Tokyo 192-0065 (Japan); Tanaka, Sachiko; Hirano, Toshihiko [Department of Clinical Pharmacology, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan); Kroetz, Deanna L. [Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, 1550 4th St, RH584E Box 2911 San Francisco, CA 94158-2911 (United States); Toyoda, Hiroo [Department of Clinical Molecular Genetics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392 (Japan)

    2011-12-15

    Arsenic trioxide (arsenite, As{sup III}) has shown a remarkable clinical efficacy, whereas its side effects are still a serious concern. Therefore, it is critical to understand the effects of As{sup III} on human-derived normal cells for revealing the mechanisms underlying these side effects. We examined the effects of As{sup III} on primary cultured chorion (C) and amnion (A) cells prepared from human fetal membranes. A significant dose-dependent As{sup III}-mediated cytotoxicity was observed in the C-cells accompanied with an increase of lactate dehydrogenase (LDH) release. Higher concentrations of As{sup III} were required for the A-cells to show cytotoxicity and LDH release, suggesting that the C-cells were more sensitive to As{sup III} than the A-cells. The expression levels of aquaporin 9 (AQP9) were approximately 2 times higher in the C-cells than those in the A-cells. Both intracellular arsenic accumulation and its cytotoxicity in the C-cells were significantly abrogated by sorbitol, a competitive AQP9 inhibitor, in a dose-dependent manner. The protein expression levels of multidrug resistance-associated protein (MRP) 2 were downregulated by As{sup III} in the C-cells, but not in the A-cells. No significant differences in the expression levels of MRP1 were observed between C- and A-cells. The protein expression of P-glycoprotein (P-gp) was hardly detected in both cells, although a detectable amount of its mRNA was observed. Cyclosporine A, a broad-spectrum inhibitor for ABC transporters, and MK571, a MRP inhibitor, but not PGP-4008, a P-gp specific inhibitor, potently sensitized both cells to As{sup III}-mediated cytotoxicity. These results suggest that AQP9 and MRP2 are involved in controlling arsenic accumulation in these normal cells, which then contribute to differential sensitivity to As{sup III} cytotoxicity between these cells. -- Highlights: Black-Right-Pointing-Pointer Examination of effect of As{sup III} on primary cultured chorion (C) and amnion

  20. Human amnion-derived mesenchymal stem cells protect against UVA irradiation-induced human dermal fibroblast senescence, in vitro

    Science.gov (United States)

    Zhang, Chunli; Yuchi, Haishen; Sun, Lu; Zhou, Xiaoli; Lin, Jinde

    2017-01-01

    The aim of the present study was to determine if human amnion-derived mesenchymal stem cells (HAMSCs) exert a protective effect on ultraviolet A (UVA) irradiation-induced human dermal fibroblast (HDF) senescence. A senescence model was constructed as follows: HDFs (104–106 cells/well) were cultured in a six-well plate in vitro and then exposed to UVA irradiation at 9 J/cm2 for 30 min. Following the irradiation period, HDFs were co-cultured with HAMSCs, which were seeded on transwells. A total of 72 h following the co-culturing, senescence-associated β-galactosidase staining was performed and reactive oxygen species (ROS) content and mitochondrial membrane potential (Δψm) were detected in the HDFs via flow cytometric analysis. The results demonstrated that the percentage of HDFs, detected via staining with X-gal, were markedly decreased when co-cultured with human HAMSCs, compared with the group that were not co-cultured. The ROS content was decreased and the mitochondrial membrane potential (Δψm) recovered in cells treated with UVA and HAMSCs, compared with that of cells treated with UVA alone. Reverse transcription-quantitative polymerase chain reaction revealed the significant effects of HAMSCs on the HDF senescence marker genes p53 and matrix metalloproteinase-1 mRNA expression. In addition to this, western blot analysis verified the effects of HAMSCs on UVA induced senescence, providing a foundation for novel regenerative therapeutic methods. Furthermore, the results suggested that activation of the extracellular-signal regulated kinase 1/2 mitogen activated protein kinase signal transduction pathway, is essential for the HAMSC-mediated UVA protective effects. The decrease in ROS content additionally indicated that HAMSCs may exhibit the potential to treat oxidative stress-mediated UVA skin senescence in the future. PMID:28627622

  1. Comparative study of regenerative effects of mesenchymal stem cells derived from placental amnion, chorion and umbilical cord on dermal wounds.

    Science.gov (United States)

    Ertl, Juliane; Pichlsberger, Melanie; Tuca, Alexandru-Cristian; Wurzer, Paul; Fuchs, Jakob; Geyer, Stefan H; Maurer-Gesek, Barbara; Weninger, Wolfgang J; Pfeiffer, Dagmar; Bubalo, Vladimir; Parvizi, Daryousch; Kamolz, Lars-Peter; Lang, Ingrid

    2018-05-01

    Mesenchymal stem/stromal cells derived from human term placentas (PMSCs) are novel therapeutic agents and more topical than ever. Here we evaluated the effects of three types of PMSCs on wound healing in an in vivo mouse model: Amnion-derived MSCs (AMSCs), blood vessel-derived MSCs (BV-MSCs) from the chorionic plate and Wharton's jelly-derived MSCs (WJ-MSCs) from the umbilical cord. We topically applied PMSCs onto skin wounds in mice using the dermal substitute Matriderm ® as carrier and evaluated wound healing parameters. In addition, we investigated the effects of all PMSC types under co-application with placental endothelial cells (PLECs). After 8 days, we compared the percent of wound closure and the angiogenic potential between all groups. AMSCs, BV-MSCs and WJ-MSCs significantly induced a faster healing and a higher number of blood vessels in the wound when compared to controls (Matriderm ® -alone). PLECs did not further improve the advantageous effects of PMSC-treatment. Quantitative data and 3D analysis by high resolution episcopic microscopy confirmed a lower density of vessels in Matriderm ® /PMSCs/PLECs co-application compared to Matriderm ® /PMSCs treatment. Results indicate that all three PMSC types exert similar beneficial effects on wound closure and neovascularization in our mouse model. Using Matriderm ® as carrier for PMSCs propagates rapid cell migration towards the wound area that allows a fast and clinically practicable method for stem cell application. These promising effects warrant further investigation in clinical trials. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

  2. Self-organized amniogenesis by human pluripotent stem cells in a biomimetic implantation-like niche

    Science.gov (United States)

    Shao, Yue; Taniguchi, Kenichiro; Gurdziel, Katherine; Townshend, Ryan F.; Xue, Xufeng; Yong, Koh Meng Aw; Sang, Jianming; Spence, Jason R.; Gumucio, Deborah L.; Fu, Jianping

    2017-04-01

    Amniogenesis--the development of amnion--is a critical developmental milestone for early human embryogenesis and successful pregnancy. However, human amniogenesis is poorly understood due to limited accessibility to peri-implantation embryos and a lack of in vitro models. Here we report an efficient biomaterial system to generate human amnion-like tissue in vitro through self-organized development of human pluripotent stem cells (hPSCs) in a bioengineered niche mimicking the in vivo implantation environment. We show that biophysical niche factors act as a switch to toggle hPSC self-renewal versus amniogenesis under self-renewal-permissive biochemical conditions. We identify a unique molecular signature of hPSC-derived amnion-like cells and show that endogenously activated BMP-SMAD signalling is required for the amnion-like tissue development by hPSCs. This study unveils the self-organizing and mechanosensitive nature of human amniogenesis and establishes the first hPSC-based model for investigating peri-implantation human amnion development, thereby helping advance human embryology and reproductive medicine.

  3. Radiosensitivity of angiogenic and mitogenic factors in human amniotic membrane

    International Nuclear Information System (INIS)

    Deocaris, Custer C.; De Guzman, Zenaida M.; Deocaris, Chester C.; Jacinto, Sonia D.

    2003-01-01

    Amniotic membrane as a temporary biological dressing remains as a beneficial and cost-effective means of treating burns in developing countries. This medical application is attributed mainly to placental structural and biochemical features that are important for maintaining proper embryonic development. Since fresh amnions are nevertheless for straightforward clinical use and for preservation, radiation-sterilization is been performed to improve the safety of this placental material. However, like any other sterilization method, gamma-radiation may induce physical and chemical changes that may influence the biological property of the material. Thus, the aim of this study is to compare the effects of various levels of radiation-sterilization protocols for human amnions on angiogenic (neovascularization) and epithelial-mitogenic activities, both of which are physiological processes fundamental to wound healing. Water-soluble extract of non-irradiated amnions demonstrates a strong stimulatory effect on both cell proliferation and angiogenesis. No change in biological activity is seen in amnions irradiated at 25 kGy, the sterilization dose used by the Philippine Nuclear Research Institute (PNRI) for the production of radiation-sterilized human amniotic membranes (RSHAM). However, it appears that amniotic angiogenic factors are more radiosensitive than its mitogenic components, evident from the depressed vascularization of the chorioallantoic membrane (CAM) exposed to 35 kGy-irradiated amnions. The dose of 35 kGy is at present the medical sterilization dose used at the Central Tissue Bank in Warsaw (Poland) for the preparation of their amnion allografts. (Authors)

  4. Soft Tissue Repair with Easy-Accessible Autologous Newborn Placenta or Umbilical Cord Blood in Severe Malformations: A Primary Evaluation

    Science.gov (United States)

    2017-01-01

    Disrupted organogenesis leads to permanent malformations that may require surgical correction. Autologous tissue grafts may be needed in severe lack of orthotopic tissue but include donor site morbidity. The placenta is commonly discarded after birth and has a therapeutic potential. The aim of this study was to determine if the amnion from placenta or plasma rich of growth factors (PRGF) with mononuclear cells (MNC) from umbilical cord blood (UCB), collected noninvasively, could be used as bio-constructs for autologous transplantation as an easy-accessible no cell culture-required method. Human amnion and PRGF gel were isolated and kept in culture for up to 21 days with or without small intestine submucosa (SIS). The cells in the constructs showed a robust phenotype without induced increased proliferation (Ki67) or apoptosis (caspase 3), but the constructs showed decreased integrity of the amnion-epithelial layer at the end of culture. Amnion-residing cells in the SIS constructs expressed CD73 or pan-cytokeratin, and cells in the PRGF-SIS constructs expressed CD45 and CD34. This study shows that amnion and UCB are potential sources for production of autologous grafts in the correction of congenital soft tissue defects. The constructs can be made promptly after birth with minimal handling or cell expansion needed. PMID:29403534

  5. Soft Tissue Repair with Easy-Accessible Autologous Newborn Placenta or Umbilical Cord Blood in Severe Malformations: A Primary Evaluation

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    Åsa Ekblad

    2017-01-01

    Full Text Available Disrupted organogenesis leads to permanent malformations that may require surgical correction. Autologous tissue grafts may be needed in severe lack of orthotopic tissue but include donor site morbidity. The placenta is commonly discarded after birth and has a therapeutic potential. The aim of this study was to determine if the amnion from placenta or plasma rich of growth factors (PRGF with mononuclear cells (MNC from umbilical cord blood (UCB, collected noninvasively, could be used as bio-constructs for autologous transplantation as an easy-accessible no cell culture-required method. Human amnion and PRGF gel were isolated and kept in culture for up to 21 days with or without small intestine submucosa (SIS. The cells in the constructs showed a robust phenotype without induced increased proliferation (Ki67 or apoptosis (caspase 3, but the constructs showed decreased integrity of the amnion-epithelial layer at the end of culture. Amnion-residing cells in the SIS constructs expressed CD73 or pan-cytokeratin, and cells in the PRGF-SIS constructs expressed CD45 and CD34. This study shows that amnion and UCB are potential sources for production of autologous grafts in the correction of congenital soft tissue defects. The constructs can be made promptly after birth with minimal handling or cell expansion needed.

  6. A multicenter, randomized, controlled clinical trial evaluating the use of dehydrated human amnion/chorion membrane allografts and multilayer compression therapy vs. multilayer compression therapy alone in the treatment of venous leg ulcers.

    Science.gov (United States)

    Serena, Thomas E; Carter, Marissa J; Le, Lam T; Sabo, Matthew J; DiMarco, Daniel T

    2014-01-01

    Venous leg ulcers produce significant clinical and economic burdens on society and often require advanced wound therapy. The purpose of this multicenter, randomized, controlled study is to evaluate the safety and efficacy of one or two applications of dehydrated human amnion/chorion membrane allograft and multilayer compression therapy vs. multilayer compression therapy alone in the treatment of venous leg ulcers. The primary study outcome was the proportion of patients achieving 40% wound closure at 4 weeks. Of the 84 participants enrolled, 53 were randomized to receive allograft and 31 were randomized to the control group of multilayer compression therapy alone. At 4 weeks, 62% in the allograft group and 32% in the control group showed a greater than 40% wound closure (p = 0.005), thus showing a significant difference between the allograft-treated groups and the multilayer compression therapy alone group at the 4-week surrogate endpoint. After 4 weeks, wounds treated with allograft had reduced in size a mean of 48.1% compared with 19.0% for controls. Venous leg ulcers treated with allograft had a significant improvement in healing at 4 weeks compared with multilayer compression therapy alone. © 2014 by the Wound Healing Society.

  7. Amniotic membrane seeded with mesenchymal adipose-derived stem cell for coverage of wound in third degree burn: An experimental study

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    Mohammad Javad Fatemi

    2014-09-01

    Conclusion: Acellular amnion seeded with adipose-derived stem cell can result in faster wound healing and better histopathology characteristic. The amnion as a scaffold and the fat derived stem cells as healing accelerator are recommended for coverage of the 3rd degree burn wounds after excision and it may reduce the need for skin graft.

  8. The Role of Progesterone and a Novel Progesterone Receptor, Progesterone Receptor Membrane Component 1, in the Inflammatory Response of Fetal Membranes to Ureaplasma parvum Infection.

    Directory of Open Access Journals (Sweden)

    Liping Feng

    Full Text Available Ureaplasma parvum (U. parvum is gaining recognition as an important pathogen for chorioamnionitis and preterm premature rupture of membranes. We aimed to investigate the roles of progesterone (P4 and a novel progesterone receptor, progesterone receptor membrane component 1 (PGRMC1, in the response of fetal membranes to U. parvum. Fetal membrane cells (amnion, chorion and decidua were isolated and confirmed to be free of Mycoplasmataceae. Cells were treated with U. parvum (5x106 CFU, and adherence was quantified by qPCR. Amnion and chorion cells were transfected with scrambled siRNA or validated PGRMC1 siRNA for 72h. Cells were then treated with U. parvum for 4h with or without pretreatment with P4 (10-7 M or ethanol for 1h. Interleukin-8 (IL-8, matrix metalloproteinase 9 (MMP9 and cyclooxygenase (COX-2 mRNA expression were quantified by qRT-PCR. Culture medium was harvested and analyzed for IL-8 and prostaglandin (PGE2 secretion by ELISA and MMP9 activity by zymography. U. parvum had a mean adherence of 15.0±0.6%, 16.9± 3.7% and 4.7±0.3% in cultured amnion, chorion and decidua cells, respectively. Exposure to U. parvum elicited significant inflammatory responses including induction of IL-8, COX-2, PGE2 and MMP9. A possible role of PGRMC1 was identified in the inhibition of U. parvum-stimulated COX-2 and MMP9 mRNA expression in chorion cells and MMP9 activity in amnion cells. On the other hand, it might enhance the U. parvum-stimulated IL-8 protein secretion in amnion cells. P4, mediated through PGRMC1, significantly inhibited U. Parvum-induced MMP9 mRNA and COX-2 mRNA expression in chorion cells. P4 appeared to attenuate U. parvum induced IL-8 mRNA expression in chorion cells, but this P4 effect might not mediated through PGRMC1. In summary, U. parvum preferentially adheres to and induces inflammatory responses in chorion and amnion cells. P4 and PGRMC1 appear to differentially modulate the inflammatory responses induced by U. parvum among

  9. Stem cell therapy to protect and repair the developing brain: a review of mechanisms of action of cord blood and amnion epithelial derived cells

    Directory of Open Access Journals (Sweden)

    Margie eCastillo-Melendez

    2013-10-01

    Full Text Available In the research, clinical and wider community there is great interest in the use of stem cells to reduce the progression, or indeed repair brain injury. Perinatal brain injury may result from acute or chronic insults sustained during fetal development, during the process of birth, or in the newborn period. The most readily identifiable outcome of perinatal brain injury is cerebral palsy, however this is just one consequence in a spectrum of mild to severe neurological deficits. As we review, there are now clinical trials taking place worldwide targeting cerebral palsy with stem cell therapies. It will likely be many years before strong evidence-based results emerge from these trials. With such trials underway, it is both appropriate and timely to address the physiological basis for the efficacy of stem-like cells in preventing damage to, or regenerating, the newborn brain. Appropriate experimental animal models are best placed to deliver this information. Cell availability, the potential for immunological rejection, ethical and logistical considerations, together with the propensity for native cells to form terratomas, make it unlikely that embryonic or fetal stem cells will be practical. Fortunately, these issues do not pertain to the use of human amnion epithelial cells (hAECs, or umbilical cord blood (UCB stem cells that are readily and economically obtained from the placenta and umbilical cord discarded at birth. These cells have the potential for transplantation to the newborn where brain injury is diagnosed or even suspected. We will explore the novel characteristics of hAECs and undifferentiated UCB cells, as well as UCB-derived endothelial progenitor cells and mesenchymal stem cells, and how immunomodulation and anti-inflammatory properties are principal mechanisms of action that are common to these cells, and which in turn may ameliorate the cerebral hypoxia and inflammation that are final pathways in the pathogenesis of perinatal brain

  10. Role of human amnion-derived mesenchymal stem cells in promoting osteogenic differentiation by influencing p38 MAPK signaling in lipopolysaccharide -induced human bone marrow mesenchymal stem cells

    International Nuclear Information System (INIS)

    Wang, Yuli; Wu, Hongxia; Shen, Ming; Ding, Siyang; Miao, Jing; Chen, Ning

    2017-01-01

    Periodontitis is a chronic inflammatory disease induced by bacterial pathogens, which not only affect connective tissue attachments but also cause alveolar bone loss. In this study, we investigated the anti-inflammatory effects of Human amnion-derived mesenchymal stem cells (HAMSCs) on human bone marrow mesenchymal stem cells (HBMSCs) under lipopolysaccharide (LPS)-induced inflammatory conditions. Proliferation levels were measured by flow cytometry and immunofluorescence staining of 5-ethynyl-2′-deoxyuridine (EdU). Osteoblastic differentiation and mineralization were investigated using chromogenic alkaline phosphatase activity (ALP) activity substrate assays, Alizarin red S staining, and RT-PCR analysis of HBMSCs osteogenic marker expression. Oxidative stress induced by LPS was investigated by assaying reactive oxygen species (ROS) level and superoxide dismutase (SOD) activity. Here, we demonstrated that HAMSCs increased the proliferation, osteoblastic differentiation, and SOD activity of LPS-induced HBMSCs, and down-regulated the ROS level. Moreover, our results suggested that the activation of p38 MAPK signal transduction pathway is essential for reversing the LPS-induced bone-destructive processes. SB203580, a selective inhibitor of p38 MAPK signaling, significantly suppressed the anti-inflammatory effects in HAMSCs. In conclusion, HAMSCs show a strong potential in treating inflammation-induced bone loss by influencing p38 MAPK signaling. - Highlights: • LPS inhibites osteogenic differentiation in HBMSCs via suppression of p38 MAPK signaling pathway. • HAMSCs promote LPS-induced HBMSCs osteogenic differentiation through p38 MAPK signaling pathway. • HAMSCs reverse LPS-induced oxidative stress in LPS-induced HBMSCs through p38 MAPK signaling pathway.

  11. Studies of hemidesmosomes in human amnion: the use of a detergent extraction protocol for compositional and ultrastructural analysis and preparation of a hemidesmosome-enriched fraction from tissue.

    Science.gov (United States)

    Behzad, F; Jones, C J; Ball, S; Alvares, T; Aplin, J D

    1995-01-01

    A method is described for the sequential detergent and high ionic strength extraction of human amnion with the progressive enrichment of the intermediate filament (IF) cytoskeleton and its associated structures including hemidesmosomes (HD). TEM of the extracted epithelium in situ reveals IF bundles beneath the apical cell surface, around the nucleus and at the lateral edges of the cells where association with desmosomes occurs. IF bundles are also very prominent within basal cell processes where they loop through the cytoplasm adjacent to the HDs. A novel connecting filament network is observed running between the IFs and the hemidesmosomal dense plaque. The adjacent IF network contains both cytokeratin and vimentin, the latter revealed much more fully as a result of the extraction protocol. The hemidesmosomal plasma membrane contains integrin subunits alpha 6 and beta 4 and these are quantitatively retained as the basal cell surface during extraction, while nonjunctional plasma membrane is solubilised. Integrin beta 1 is found at the basolateral cell surface but, like actin, is extracted quantitatively and is not present in HDs. The extracted epithelial cells may be recovered by scraping and the IF network depolymerised to produce a particulate fraction containing short residual IFs, associated thin filaments and plaque material. This fraction contains immunoreactive cytokeratin and vimentin. Integrin alpha 6 beta 4 has been used as a biochemical criterion of the presence of HD material in the fraction. Both subunits are highly enriched. The fraction also contains the hemidesmosomal components HD1, BP230 and BP180. This method is likely to be useful in further characterisation of the HD.

  12. Potential antitumor therapeutic strategies of human amniotic membrane and amniotic fluid-derived stem cells.

    Science.gov (United States)

    Kang, N-H; Hwang, K-A; Kim, S U; Kim, Y-B; Hyun, S-H; Jeung, E-B; Choi, K-C

    2012-08-01

    As stem cells are capable of self-renewal and can generate differentiated progenies for organ development, they are considered as potential source for regenerative medicine and tissue replacement after injury or disease. Along with this capacity, stem cells have the therapeutic potential for treating human diseases including cancers. According to the origins, stem cells are broadly classified into two types: embryonic stem cells (ESCs) and adult stem cells. In terms of differentiation potential, ESCs are pluripotent and adult stem cells are multipotent. Amnion, which is a membranous sac that contains the fetus and amniotic fluid and functions in protecting the developing embryo during gestation, is another stem cell source. Amnion-derived stem cells are classified as human amniotic membrane-derived epithelial stem cells, human amniotic membrane-derived mesenchymal stem cells and human amniotic fluid-derived stem cells. They are in an intermediate stage between pluripotent ESCs and lineage-restricted adult stem cells, non-tumorigenic, and contribute to low immunogenicity and anti-inflammation. Furthermore, they are easily available and do not cause any controversial issues in their recovery and applications. Not only are amnion-derived stem cells applicable in regenerative medicine, they have anticancer capacity. In non-engineered stem cells transplantation strategies, amnion-derived stem cells effectively target the tumor and suppressed the tumor growth by expressing cytotoxic cytokines. Additionally, they also have a potential as novel delivery vehicles transferring therapeutic genes to the cancer formation sites in gene-directed enzyme/prodrug combination therapy. Owing to their own advantageous properties, amnion-derived stem cells are emerging as a new candidate in anticancer therapy.

  13. Methylation of Promoter Regions of Genes of the Human Intrauterine Renin Angiotensin System and Their Expression

    Directory of Open Access Journals (Sweden)

    Shane D. Sykes

    2015-01-01

    Full Text Available The intrauterine renin angiotensin system (RAS is implicated in placentation and labour onset. Here we investigate whether promoter methylation of RAS genes changes with gestation or labour and if it affects gene expression. Early gestation amnion and placenta were studied, as were term amnion, decidua, and placenta collected before labour (at elective caesarean section or after spontaneous labour and delivery. The expression and degree of methylation of the prorenin receptor (ATP6AP2, angiotensin converting enzyme (ACE, angiotensin II type 1 receptor (AGTR1, and two proteases that can activate prorenin (kallikrein, KLK1, and cathepsin D, CTSD were measured by qPCR and a DNA methylation array. There was no effect of gestation or labour on the methylation of RAS genes and CTSD. Amnion and decidua displayed strong correlations between the percent hypermethylation of RAS genes and CTSD, suggestive of global methylation. There were no correlations between the degree of methylation and mRNA abundance of any genes studied. KLK1 was the most methylated gene and the proportion of hypermethylated KLK1 alleles was lower in placenta than decidua. The presence of intermediate methylated alleles of KLK1 in early gestation placenta and in amnion after labour suggests that KLK1 methylation is uniquely dynamic in these tissues.

  14. Amniotic Mesenchymal Stromal Cells Exhibit Preferential Osteogenic and Chondrogenic Differentiation and Enhanced Matrix Production Compared With Adipose Mesenchymal Stromal Cells.

    Science.gov (United States)

    Topoluk, Natasha; Hawkins, Richard; Tokish, John; Mercuri, Jeremy

    2017-09-01

    Therapeutic efficacy of various mesenchymal stromal cell (MSC) types for orthopaedic applications is currently being investigated. While the concept of MSC therapy is well grounded in the basic science of healing and regeneration, little is known about individual MSC populations in terms of their propensity to promote the repair and/or regeneration of specific musculoskeletal tissues. Two promising MSC sources, adipose and amnion, have each demonstrated differentiation and extracellular matrix (ECM) production in the setting of musculoskeletal tissue regeneration. However, no study to date has directly compared the differentiation potential of these 2 MSC populations. To compare the ability of human adipose- and amnion-derived MSCs to undergo osteogenic and chondrogenic differentiation. Controlled laboratory study. MSC populations from the human term amnion were quantified and characterized via cell counting, histologic assessment, and flow cytometry. Differentiation of these cells in comparison to commercially purchased human adipose-derived mesenchymal stromal cells (hADSCs) in the presence and absence of differentiation media was evaluated via reverse transcription polymerase chain reaction (PCR) for bone and cartilage gene transcript markers and histology/immunohistochemistry to examine ECM production. Analysis of variance and paired t tests were performed to compare results across all cell groups investigated. The authors confirmed that the human term amnion contains 2 primary cell types demonstrating MSC characteristics-(1) human amniotic epithelial cells (hAECs) and (2) human amniotic mesenchymal stromal cells (hAMSCs)-and each exhibited more than 90% staining for MSC surface markers (CD90, CD105, CD73). Average viable hAEC and hAMSC yields at harvest were 2.3 × 10 6 ± 3.7 × 10 5 and 1.6 × 10 6 ± 4.7 × 10 5 per milliliter of amnion, respectively. As well, hAECs and hAMSCs demonstrated significantly greater osteocalcin ( P = .025), aggrecan ( P

  15. Increased oxidative stress in human fetal membranes overlying the cervix from term non-labouring and post labour deliveries.

    Science.gov (United States)

    Chai, M; Barker, G; Menon, R; Lappas, M

    2012-08-01

    Enzymatic breakdown of the collagen-rich extracellular matrix (ECM) that connects the amnion and chorion layers of the fetal membranes is one of the key events leading to rupture of membranes. Oxidant stress caused by increased formation of reactive oxygen species and/or reduced antioxidant capacity may predispose to membrane rupture, a major cause of preterm birth. The aim of this study was to determine the effect of human labour and supracervical (SC) apposition on antioxidant enzymes and 8-isoprostane (a marker of lipid peroxidation). To determine the effect of human labour on oxidative stress status, fetal membranes from the SC site (SCS) were collected from women at term Caesarean section (no labour), and from the site of membrane rupture (SOR) after spontaneous labour onset and delivery (post labour). To determine the effect of SC apposition on oxidative stress status, amnion was collected from the SCS and a distal site (DS) in women at term Caesarean section in the absence of labour. The release of 8-isoprostane was significantly higher in amnion from the SCS compared to DS, and in fetal membranes from the SOR compared to the SCS. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity were lower in amnion from the SC compared to DS. SOD gene expression and enzyme activity were lower in fetal membranes after labour. There was no difference in expression or activity in catalase, GPx and glutathione reductase (GSR) between no labour and post labour fetal membranes. In primary amnion cells, SOD supplementation significantly augmented IL-1β induced MMP-9 expression and activity. In summary, non-labouring SC fetal membranes are characterised by reduced antioxidant enzyme activity when compared to distal membranes, and, as such, may be more susceptible to oxidative damage and thus membrane rupture. Copyright © 2012 Elsevier Ltd. All rights reserved.

  16. In vitro secretion profiles of interleukin (IL-1beta, IL-6, IL-8, IL-10, and TNF alpha after selective infection with Escherichia coli in human fetal membranes

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    Maida-Claros Rolando

    2007-12-01

    Full Text Available Abstract Background Chorioamniotic membranes infection is a pathologic condition in which an abnormal secretion of proinflammatory cytokines halts fetal immune tolerance. The aim of the present study was to evaluate the functional response of human chorioamniotic membranes, as well as the individual contribution of the amnion and choriodecidua after stimulation with Escherichia coli, a pathogen associated with preterm labor. Methods Explants of chorioamniotic membranes from 10 women (37–40 weeks of gestation were mounted and cultured in a Transwell system, which allowed us to test the amnion and choriodecidua compartments independently. Escherichia coli (1 × 10 6 CFU/mL was added to either the amniotic or the choriodecidual regions or both; after a 24-h incubation, the secretion of IL-1beta, IL-6, TNFalpha, IL-8, and IL-10 in both compartments was measured using a specific ELISA. Data were analyzed by Kruskal-Wallis one-way analysis of variance. Results After stimulation with Escherichia coli, the choriodecidua compartment showed an increase in the secretion of IL-1beta (21-fold, IL-6 (2-fold, IL-8 (6-fold, and IL-10 (37-fold, regardless of which side of the membrane was stimulated; TNFalpha secretion augmented (22-fold also but only when the stimulus was applied simultaneously to both sides. When the amnion was stimulated directly, the level of IL-1beta (13-fold rose significantly; however, the increase in IL-8 secretion was larger (20-fold, regardless of the primary site of infection. TNFalpha secretion in the amnion compartment rose markedly only when Escherichia coli was simultaneously applied to both sides. Conclusion Selective stimulation of fetal membranes with Escherichia coli results in a differential production of IL-1beta, IL-6, TNFalpha, IL-8, and IL-10. These tissues were less responsive when the amnion side was stimulated. This is in agreement with the hypothesis that the choriodecidua may play a primary role during an ascending

  17. Grafting with Cryopreserved Amniotic Membrane versus Conservative Wound Care in Treatment of Pressure Ulcers: A Randomized Clinical Trial.

    Science.gov (United States)

    Dehghani, Mehdi; Azarpira, Negar; Mohammad Karimi, Vahid; Mossayebi, Hamid; Esfandiari, Elaheh

    2017-10-01

    To compare the healing process of pressure ulcers treated with cryopreserved human amniotic membrane allograft and routine pressure ulcer care in our hospital. From January 2012 to December 2013, in a prospective randomized clinical trial (IRCT201612041335N2), 24 patients with second and third stage of pressure ulcers were enrolled in this study. All patients needed split-thickness skin grafts for pressure ulcer-wound coverage. Selected patients had symmetric ulcers on both upper and lower extremities. The patients were randomly divided into two groups: amnion and control. In the amnion group, the ulcer was covered with cryopreserved amniotic membrane and in the control group it was treated with local Dilantin powder application. The duration and success rate of complete healing was compared between the two groups. The study group was composed of 24 pressure ulcers in 24 patients (19 males and 5 females) with a mean age of 44±12.70 years. The demographic characteristics, ulcer area, and underlying diseases were similar in both groups. The early sign of response, such as decrease in wound discharge, was detected 12-14 days after biological dressing. Complete pressure ulcer healing occurred only in the amnion group ( p pressure ulcers.

  18. Using Amniotic Membrane as Wound Covering After Cesarean Section Operation

    International Nuclear Information System (INIS)

    Manjas, Menkher; Helmi, Helfial

    2002-01-01

    Early mobilization and good wound operation healing are the other aim of all treatment for cesarean section operation. Especially for wound healing we can use amniotic membrane which is soft, easy to shape wound surface, satisfactory adhesive properties, good elasticity and sufficient transparency which allows wound control without secondary redressing. From July 1999 until December 1999 total of 196 patients undergoing cesarean section with amnion as would covering were evaluated for injection of amnion, sign of wound injection, and duration of wound healing. Amniotic membrane gives best results in wound healing, no sing of rejection and there is no different results between emergency operation and elective operation, clean and dirty operation

  19. Human Amniotic Membrane Dressing: an Excellent Method for Outpatient Management of Burn Wounds

    Directory of Open Access Journals (Sweden)

    Ali Akbar Mohammadi

    2009-03-01

    Full Text Available Background: Burns are among the most common traumas indeveloping countries, which consume large amounts of medicalresources. It is important to find an appropriate materialfor dressing of burn wounds that improves healing and is readilyavailable, easily applicable, and economical.Methods: In a single-blind randomized controlled clinicaltrial from March to October 2006, 211 patients with less than20% burn were enrolled into two groups. The first group contained104 patients with average burn of 11.90± 3.80% of totalbody surface area (TBSA for whom amnion dressing wasused. The second group composed of 107 patients with averageburn of 12.30± 4.14% of TBSA treated with routine silversulfadiazine dressing.Results: Amniotic membrane usage was accompanied by accelerationin wound healing, less need for skin graft, and lesspain. The mean healing time in superficial parts of burnwounds in the amnion group was significantly shorter than thecontrol group (9.50±2.13 v 14.30±2.60 days; P value < 0.01.The extent of the wound with granulation tissue which neededskin graft was less in the amnion group (2.10 ± 2.21% v 4.20±1.44%; P value < 0.01.Conclusion: Widespread use of amniotic membrane dressingis recommended for limited burn wound management.

  20. Biomechanical assays amniotic membrane preserved in glycerol correlating with optical coherence tomography (OCT) and thermal gravimetric analysis (TG)

    International Nuclear Information System (INIS)

    Soares, Fernando Augusto N.; Santin, Stefany P.; Martino Junior, Antonio C.; Machado, Luci Diva B.; Freitas, Anderson Z.; Mathor, Monica B.

    2013-01-01

    Amnion or amniotic membranes (AM) are interchangeable terms used in the literature being internal part of the fetal membranes, non-vascular and multicellular tissue. The amnion has been widely used as a graft ophthalmic surgical as well as carrier substrate stem cell tissue equivalent for ocular surface reconstruction. The AM reduces scar formation and inflammation on the ocular surface, promotes epithelization also been used as a biological bandage covering the wound or burns, reducing dehydration and allowing regeneration of these areas. The amnion has usually 0.02 to 0.5 mm thick and consists of five subsequent layers: epithelium, basement membrane, compact layer, fibroblast layer and spongy layer. The mechanical strength from the membrane structure as well as the elasticity are factors attractive to the use of amnion as a surgical graft. Higher levels of rigidity and strength may improve the graft resistance necessary to resist the stress induced during growth of the new tissue formed. The amniotic membrane is obtained at elective caesarean section and subsequently, under sterile conditions, sectioned and separated from chorion and placenta, and free blood clots. The serological tests are done at the time of collection of tissue and 6 months after delivery to confirm the results. There are different methods for storing MA in tissue banks as fresh, high concentrations of glycerol, among others. The use of fresh membrane has some limitations due to the need to rapid use and high risk of contamination, however the amniotic membrane in glycerol has antiviral and antibacterial property which is dependent on the concentration, time and temperature. The AM used in transplants must be sterile to prevent the transmission of any disease. Although sterilization by radiation is an effective procedure, it can interfere on the membrane structure. Thus, verification of potential changes caused by ionizing radiation in amnion was made using the tensile test by calculating the

  1. Biomechanical assays amniotic membrane preserved in glycerol correlating with optical coherence tomography (OCT) and thermal gravimetric analysis (TG)

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Fernando Augusto N.; Santin, Stefany P.; Martino Junior, Antonio C.; Machado, Luci Diva B.; Freitas, Anderson Z.; Mathor, Monica B., E-mail: fernandonevessoares@yahoo.com.br [Instituto de Pesquisas Energetias Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2013-07-01

    Amnion or amniotic membranes (AM) are interchangeable terms used in the literature being internal part of the fetal membranes, non-vascular and multicellular tissue. The amnion has been widely used as a graft ophthalmic surgical as well as carrier substrate stem cell tissue equivalent for ocular surface reconstruction. The AM reduces scar formation and inflammation on the ocular surface, promotes epithelization also been used as a biological bandage covering the wound or burns, reducing dehydration and allowing regeneration of these areas. The amnion has usually 0.02 to 0.5 mm thick and consists of five subsequent layers: epithelium, basement membrane, compact layer, fibroblast layer and spongy layer. The mechanical strength from the membrane structure as well as the elasticity are factors attractive to the use of amnion as a surgical graft. Higher levels of rigidity and strength may improve the graft resistance necessary to resist the stress induced during growth of the new tissue formed. The amniotic membrane is obtained at elective caesarean section and subsequently, under sterile conditions, sectioned and separated from chorion and placenta, and free blood clots. The serological tests are done at the time of collection of tissue and 6 months after delivery to confirm the results. There are different methods for storing MA in tissue banks as fresh, high concentrations of glycerol, among others. The use of fresh membrane has some limitations due to the need to rapid use and high risk of contamination, however the amniotic membrane in glycerol has antiviral and antibacterial property which is dependent on the concentration, time and temperature. The AM used in transplants must be sterile to prevent the transmission of any disease. Although sterilization by radiation is an effective procedure, it can interfere on the membrane structure. Thus, verification of potential changes caused by ionizing radiation in amnion was made using the tensile test by calculating the

  2. Case report

    African Journals Online (AJOL)

    abp

    2015-11-19

    Nov 19, 2015 ... There was no history of rupture of amnion membranes. Fetal heart ... aggressive hydration therapy including blood products, by central venous catheterization, she had persistant bradycardia and hypotension. Vasopressin ...

  3. Clinical application of amniotic membranes on a patient with epidermolysis bullosa.

    Science.gov (United States)

    Martínez Pardo, M E; Reyes Frías, M L; Ramos Durón, L E; Gutiérrez Salgado, E; Gómez, J C; Marín, M A; Luna Zaragoza, D

    1999-01-01

    The case of a patient with dystrophic epidermolysis bullosa treated with radiosterilised amniotic membranes is presented. The disorder is a congenital disease characterised by a poor desmosomal junction in the keratinocyte membrane. After proper donor screening, amnios were collected at Hospital Central Sur de Alta Especialidad (HCSAE), PEMEX and microbiological analysis was performed at Universidad Nacional Autónoma de México, FQUNAM, (Biology Dept. of the Chemistry Faculty, National Autonomous University of Mexico), before and after radiation sterilisation. Processing, packaging and sterilisation were performed at Instituto Nacional de Investigaciones Nucleares, ININ, (National Nuclear Research Institute). The patient, a ten-year-old boy with severe malnutrition, extensive loss of skin and pseudomonad infection in the whole body, was treated with gentle debridement in a Hubbard bath. Later amnion application was performed with sterilised amnios by using two different processes, in one of which the amnion was sterilised with paracetic acid, preserved in glycerol, kindly donated by the German Institute for Tissue and Cell Replacement and applied by Dr. Johannes C. Bruck, IAEA visiting expert, and the other amnion was processed at ININ: air dried and sterilised by gamma radiation at dose of 30 kGy. After spontaneous epithelisation was successfully promoted for seven days, the pain was alleviated and mobility was improved in a few hours and the patient's general condition was so improved that in a month he was discharged. Unfortunately, because this disease is revertive and has malignant degeneration, the prognosis is not good.

  4. Thymic size at birth in preterm infants with severe respiratory ...

    African Journals Online (AJOL)

    To determine whether the thymic size in preterm infants with severe respiratory distress syndrome (RDS) can be used to predict ... chorio-amnionitis is associated with a small thymus at birth.4,7. In those ..... Acta Paediatr 2000; 89: 975-978. 6.

  5. Inhibition of PIM1 kinase attenuates inflammation-induced pro-labour mediators in human foetal membranes in vitro.

    Science.gov (United States)

    Lim, Ratana; Barker, Gillian; Lappas, Martha

    2017-06-01

    Does proviral integration site for Moloney murine leukaemic virus (PIM)1 kinase play a role in regulating the inflammatory processes of human labour and delivery? PIM1 kinase plays a critical role in foetal membranes in regulating pro-inflammatory and pro-labour mediators. Infection and inflammation have strong causal links to preterm delivery by stimulating pro-inflammatory cytokines and collagen degrading enzymes, which can lead to rupture of membranes. PIM1 has been shown to have a role in immune regulation and inflammation in non-gestational tissues; however, its role has not been explored in the field of human labour. PIM1 expression was analysed in myometrium and/or foetal membranes obtained at term and preterm (n = 8-9 patients per group). Foetal membranes, freshly isolated amnion cells and primary myometrial cells were used to investigate the effect of PIM1 inhibition on pro-labour mediators (n = 5 patients per treatment group). Foetal membranes, from term and preterm, were obtained from non-labouring and labouring women, and from preterm pre-labour rupture of membranes (PPROM) (n = 9 per group). Amnion was collected from women with and without preterm chorioamnionitis (n = 8 per group). Expression of PIM1 kinase was determined by qRT-PCR and western blotting. To determine the effect of PIM1 kinase inhibition on the expression of pro-inflammatory and pro-labour mediators induced by bacterial products lipopolysaccharide (LPS) (10 μg/ml) and flagellin (1 μg/ml) and pro-inflammatory cytokine tumour necrosis factor (TNF) (10 ng/ml), chemical inhibitors SMI-4a (20 μM) and AZD1208 (50 μM) were used in foetal membrane explants and siRNA against PIM1 was used in primary amnion cells. Statistical significance was set at P membranes after spontaneous term labour compared to no labour at term and in amnion with preterm chorioamnionitis compared to preterm with no chorioamnionitis. There was no change in PIM1 expression with preterm labour or PPROM

  6. Evolution of the Radio-sterilized Tissues Bank of the Instituto Nacional de Investigaciones Nucleares

    International Nuclear Information System (INIS)

    Martinez P, M. E.; Reyes F, M. L.; Luna Z, D.

    2013-10-01

    The Radio-sterilized Tissues Bank (RTB) of the Instituto Nacional de Investigaciones Nucleares (ININ) was established in 1997, as national project supported by the IAEA, who provided equipment, training and expert missions. In July of 1999, the Secretaria de Salud (Mexico) granted the sanitary license to the RTB. The first radio-sterilized tissue was the amnion. Later on, the process of frozen and lyophilized pig skin was development. Both tissue types are used as biological dressings in patient with burns, ulcers or on injuries difficult to heal, the amnion is also used for the damage treatment in the ocular surface. In 2003, the ININ improved the facilities of the RTB and in August of that same year the certification of the Quality Administration System was obtained under the standard ISO 9001:2000, at the present time ISO 9001:2008. The support of the IAEA granted to Mexico was of 1997-2004 and 2009-2012, by means of regional and inter regional projects where was obtained equipment and training mainly, for personal of the ININ and other institutions. With the last project supported by the IAEA, two very important documents were generated for the tissues bank of Argentina, Brazil, Colombia, Costa Rica, Cuba, Ecuador, El Salvador, Mexico, Peru, Uruguay and Venezuela, strengthening the interaction among these countries. To offer more radio-sterilized tissues to the medical community, in June of 2007 the RTB signed and agreement with the Secretaria de Salud of the Mexico State by means of the Centro Estatal de Transplantes del Estado de Mexico, for the process of muscle-skeletal tissue, skin and amnion. At present, besides amnion and pig skin, in the RTB there is in existence powdered and chips bone; concluding the validation stage the human skin process is. As a social function of the ININ, the RTB has contributed to the health improvement of patients of more than 50 hospital institutions, mainly those that assist to patient of low resources in different cities of

  7. Evolution of the Radio-sterilized Tissues Bank of the Instituto Nacional de Investigaciones Nucleares; Evolucion del Banco de Tejidos Radioesterilizados del Instituto Nacional de Investigaciones Nucleares

    Energy Technology Data Exchange (ETDEWEB)

    Martinez P, M. E.; Reyes F, M. L.; Luna Z, D., E-mail: esther.martinez@inin.gob.mx [ININ, Carretera Mexico-Toluca s/n, 52750 Ocoyoacac, Estado de Mexico (Mexico)

    2013-10-15

    The Radio-sterilized Tissues Bank (RTB) of the Instituto Nacional de Investigaciones Nucleares (ININ) was established in 1997, as national project supported by the IAEA, who provided equipment, training and expert missions. In July of 1999, the Secretaria de Salud (Mexico) granted the sanitary license to the RTB. The first radio-sterilized tissue was the amnion. Later on, the process of frozen and lyophilized pig skin was development. Both tissue types are used as biological dressings in patient with burns, ulcers or on injuries difficult to heal, the amnion is also used for the damage treatment in the ocular surface. In 2003, the ININ improved the facilities of the RTB and in August of that same year the certification of the Quality Administration System was obtained under the standard ISO 9001:2000, at the present time ISO 9001:2008. The support of the IAEA granted to Mexico was of 1997-2004 and 2009-2012, by means of regional and inter regional projects where was obtained equipment and training mainly, for personal of the ININ and other institutions. With the last project supported by the IAEA, two very important documents were generated for the tissues bank of Argentina, Brazil, Colombia, Costa Rica, Cuba, Ecuador, El Salvador, Mexico, Peru, Uruguay and Venezuela, strengthening the interaction among these countries. To offer more radio-sterilized tissues to the medical community, in June of 2007 the RTB signed and agreement with the Secretaria de Salud of the Mexico State by means of the Centro Estatal de Transplantes del Estado de Mexico, for the process of muscle-skeletal tissue, skin and amnion. At present, besides amnion and pig skin, in the RTB there is in existence powdered and chips bone; concluding the validation stage the human skin process is. As a social function of the ININ, the RTB has contributed to the health improvement of patients of more than 50 hospital institutions, mainly those that assist to patient of low resources in different cities of

  8. Radiation dose setting for sterilization of health care items in relation to product microbiological quality

    International Nuclear Information System (INIS)

    Norimah Yusof; Nagi Marsit; Asnah Hassan

    1997-01-01

    Radiation dose of 25 k gray is no longer a generally accepted dose for sterilization. ISO document no. 11137 stated that a manufacturer can decide the dose to sterilize his product depending on the product's microbiological quality (number and type of the contaminants) and the sterility assurance level (SAL) should attain in relation to its usage. Five health care products were selected for the microbiological studies including bio burden counts, identification of most commonly found microorganisms and the radioresistance (D sub 10 value) of the selected isolates. Radiation dose was then determined by two methods, namely Method for Dose Validation of ISO 11137, and calculation based on log survival or population cycle reduction. At a given SAL of 10 sup -6 the radiation sterilization dose obtained by both methods was influenced by microbiological quality of the product. Sterilization dose set by the ISO Method I (Cotton Ball 19.4 kGy, Syringe 20.4 kGy, Suture 15. 0 kGy, Surgical Glove 24.9 kGy and Amnion 17.8 kGy) was higher than the dose calculated according to the log cycle reduction concept in all the products (Cotton Ball 14. 0 kGy, Syringe 15.5 kGy, Suture 11. 6 kGy, Surgical Glove 18. 0 kGy and Amnion 12.6 kGy). The ISO method has limitation on bio products such as amnion and other high valued products which are produced in small number with low bio burden and microorganism spectrum different from those commonly found on medical items

  9. Clinical analysis of amniotic membrane patches and grafts for acute ocular surface burn

    Directory of Open Access Journals (Sweden)

    Lin Li

    2015-01-01

    Full Text Available AIM: To investigate the effect and value of amniotic membrane patches and grafts for acute ocular surface burn at different degrees.METHODS: A retrospective analysis of 28 cases(28 eyesaffected by ocular chemical or thermal burn with different degree were included in our hospital from March 2007 to March 2012. Amniotic membrane patched was undergone in 13 eyes with fresh amnion that the patients corneal burns degree Ⅱ or Ⅲ with partial limbal buns at degree Ⅳ. Amniotic membrane grafts was performed in 15 eyes with fresh amnion that the patients all corneal burns at degree Ⅲ with the whole limbal necrosis without severe eyelid defect. The follow-up time ranged 6~24mo. The postoperative visual acuity, the condition of amniotic membrane transplant, renovation of cornea and complications were observed. RESULTS: Postoperative corrected visual acuity was improved in 20 eyes(71%, it was not changed in 5 eyes(18%, the visual acuity declined in 3 eyes(11%. The amniotic membrane survived in 23 eyes and the survival rate was up to 82%. The cornea of 4 eyes recovered to transparent, nebula emceed in 8 eyes eventually, corneal macula emerged in 10 eyes, 4 eyes ended up with leukoma, 2 eyes developed corneal melting after therapy, then received lamellar keratoplasty. Corneal surface become epithelization after amnion patches or grafts, but any of them have recurrent epithelial erosion, and become stable epithalization after repeat operation.CONCLUSION: Amniotic membrane patches and grafts is an effective method to deal with acute ocular surface burn.

  10. Chitosan Cross-linked Reconstituted Amniotic Collagen Membrane ...

    Indian Academy of Sciences (India)

    First page Back Continue Last page Overview Graphics. Chitosan Cross-linked Reconstituted Amniotic Collagen Membrane – An Excellent Cell Substratum. The KERATINOCYTE proliferation and Differentiation into multiple layers is due to the presence of type - IV collagen in the amnion. Cultured FIBROBLASTS had good ...

  11. Short Communications

    African Journals Online (AJOL)

    This method of amnion formation has been reported in only a few bat species,. e.g. the phyllostomatids and desmodontids. Both these groups have been considered exceptional among bats in exhibiting completely interstitial implantation (Hamlett 1935; Wimsatt. 1954). In members of both these groups amniogenesis by.

  12. Plazentaanatomie und -physiologie mit uteroplazentarem Kreislauf

    NARCIS (Netherlands)

    Engel, N.; Schreiber, J. U.; Kranke, Peter

    2017-01-01

    Die Plazenta besteht neben der Nabelschnur aus parenchymatösem Gewebe, der sog. Eihaut (Amnion) und der Zottenhaut (Chorion). Sie fungiert als Schnittstelle zwischen Mutter und Fetus und ermöglicht den physiologischen Transfer von Gasen, Nährstoffen und Metaboliten. Die Funktion der Plazenta ist als

  13. The vanishing twin: morphologic and cytogenetic evaluation of an ultrasonographic phenomenon

    DEFF Research Database (Denmark)

    Rudnicki, M; Vejerslev, L O; Junge, Jette

    1991-01-01

    Twin pregnancy was observed by ultrasonographic examination in the 6th week of gestation. After singleton term delivery a thickening of the membranes opposite to the main placenta showed degenerated chorionic villi embedded between one layer of amnion and chorion; no fetal parts were observed. Vi...

  14. Tissue Banking: Current procedures, ethical consideration and ...

    African Journals Online (AJOL)

    Tissue banking provides safe and effective cells and tissues for transplantation in reconstruction surgery. Bone, amnion, skin, cartilage, heart valves and xenograft tissues are the most commonly used biological tissues. Acquisition of tissue is dependent on elaborate donor screening criteria based on medical and social ...

  15. Schizamniogenesis in the rusty bat, Pipistrellus rusticus | van der ...

    African Journals Online (AJOL)

    Rusty bats are seasonally monoestrous, carrying a single foetus in each of the two uterine horns. Implantation is superficial with amniogenesis initiated very early during embryogenesis. Contrary to most other bat species where the amnion is formed by folding, it is formed by cavitation in the rusty bat.

  16. Amniogenesis in Schreiber's long-fingered bat Miniopterus ...

    African Journals Online (AJOL)

    Schreiber's long-fingered bat, Miniopterus schreibersii natalensis is seasonally monoestrous, carrying a single foetus in the right uterine horn. Implantation is superficial, the amnion being a pleuramnion. Lateral folds, originating from the ends of the caudal and cephalic folds, are the main contributors in the formation of the ...

  17. Cat amniotic membrane multipotent cells are nontumorigenic and are safe for use in cell transplantation

    Directory of Open Access Journals (Sweden)

    Vidane AS

    2014-08-01

    Full Text Available Atanasio S Vidane,1 Aline F Souza,1 Rafael V Sampaio,1 Fabiana F Bressan,2 Naira C Pieri,1 Daniele S Martins,2 Flavio V Meirelles,2 Maria A Miglino,1 Carlos E Ambrósio2 1Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil; 2Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, São Paulo, Brazil Abstract: Amnion-derived mesenchymal stem cells (AMSCs are multipotent cells with an enhanced ability to differentiate into multiple lineages. AMSCs can be acquired through noninvasive methods, and therefore are exempt from the typical ethical issues surrounding stem cell use. The objective of this study was to isolate and characterize AMSCs from a cat amniotic membrane for future application in regenerative medicine. The cat AMSCs were harvested after mechanical and enzymatic digestion of amnion. In culture medium, the cat AMSCs adhered to a plastic culture dish and displayed a fibroblast-like morphology. Immunophenotyping assays were positive for the mesenchymal stem cell-specific markers CD73 and CD90 but not the hematopoietic markers CD34, CD45, and CD79. Under appropriate conditions, the cat AMSCs differentiated into osteogenic, chondrogenic, and adipogenic cell lineages. One advantage of cat AMSCs was nonteratogenicity, assessed 4 weeks post injection of undifferentiated AMSCs into immunodeficient mice. These findings suggest that cat amniotic membranes may be an important and useful source of mesenchymal stem cells for clinical applications, especially for cell or tissue replacement in chronic and degenerative diseases. Keywords: amnion, cats, cell differentiation, fetal membranes, mesenchymal cells

  18. Inflammatory Response of Human Gestational Membranes to Ureaplasma parvum Using a Novel Dual-Chamber Tissue Explant System.

    Science.gov (United States)

    Potts, Lauren C; Feng, Liping; Seed, Patrick C; Jayes, Friederike L; Kuchibhatla, Maragatha; Antczak, Brian; Nazzal, Matthew K; Murtha, Amy P

    2016-05-01

    Preterm premature rupture of membranes (PPROM) is often associated with intra-amniotic inflammation and infection. Current understanding of the pathogenesis of PPROM includes activation of pro-inflammatory cytokines and proteolytic enzymes leading to compromise of membrane integrity. The impact of exposure to bacterial pathogens, including Ureaplasma parvum, on gestational membranes is poorly understood. Our objective was to develop a dual-chamber system to characterize the inflammatory response of gestational membranes to U. parvum in a directional nature. Full-thickness human gestational membrane explants, with either choriodecidua or amnion oriented superiorly, were suspended between two washers in a cylindrical device, creating two distinct compartments. Brilliant green dye was introduced into the top chamber to assess the integrity of the system. Tissue viability was evaluated after 72 h using a colorimetric cell proliferation assay. Choriodecidua or amnion was exposed to three doses of U. parvum and incubated for 24 h. Following treatment, media from each compartment were used for quantification of U. parvum (quantitative PCR), interleukin (IL)-8 (enzyme-linked immunosorbent assay), and matrix metalloproteinase (MMP)-2 and MMP-9 activity (zymography). We observed that system integrity and explant viability were maintained over 72 h. Dose-dependent increases in recovered U. parvum, IL-8 concentration, and MMP-2 activity were detected in both compartments. Significant differences in IL-8 concentration and MMP-9 activity were found between the choriodecidua and amnion. This tissue explant system can be used to investigate the inflammatory consequences of directional bacterial exposure for gestational membranes and provides insight into the pathogenesis of PPROM and infectious complications of pregnancy. © 2016 by the Society for the Study of Reproduction, Inc.

  19. Amniogenesis in the African yellow bat, Scotophilus dinganii | van ...

    African Journals Online (AJOL)

    The African yellow house bat, Scotophilus dinganii, was found to be seasonally monestrus, carrying a single foetus in each of the two uterine horns of the bicornuate uterus. Implantation was superficial, with amniogenesis initiated early during embryogenesis. The amnion in S. dinganii was a schizamnion, and it is deduced ...

  20. Development of Genetic Therapies for the Hemidesmosol Subtypes of Junction Epidermolysis Bullosa

    National Research Council Canada - National Science Library

    Christiano, Angela

    2003-01-01

    ... for in vitro epithelial reprogramming, in which we have begun to utilize different epithelial cell types as donor cells, in addition to keratinocytes, specifically, amnion and cornea cells. Collectively, we have shown that the use of gene delivery combined with epithelial cell-skin equivalent models, show significant promise toward developing a cellular.

  1. Classics revisited

    DEFF Research Database (Denmark)

    Carter, A M

    2018-01-01

    hybridus). He was able to get early stages before twinning occurred and show it was preceded by inversion of the germ layers. By the primitive streak stage there were separate embryonic shields and partition of the amnion. There was, however, a single exocoelom and all embryos were enclosed in a common set...

  2. Infection-Induced Thrombin Production: A Potential Novel Mechanism for Preterm Premature Rupture of Membranes (PPROM).

    Science.gov (United States)

    Feng, Liping; Allen, Terrence K; Marinello, William P; Murtha, Amy P

    2018-04-13

    Preterm premature rupture of membranes (PPROM) is a leading contributor to maternal and neonatal morbidity and mortality. Epidemiologic and experimental studies have demonstrated that thrombin causes fetal membrane weakening and subsequently PPROM. Although blood is suspected as the likely source of thrombin in fetal membranes and amniotic fluid of patients with PPROM, this has not been proven. Ureaplasma Parvum (U. parvum) is emerging as a pathogen involved in prematurity, including PPROM, but until now, prothrombin production directly induced by bacteria in fetal membranes has not been described. This study was designed to investigate whether U. parvum exposure can induce prothrombin production in fetal membranes cells. Primary fetal membrane cells (amnion epithelial, chorion trophoblast, and decidua stromal) or full-thickness fetal membrane tissue explants from elective, term, uncomplicated cesarean deliveries were harvested. Cells or tissue explants were infected with live U. parvum (1 x 10 5 , 1 x 10 6 , or 1 x 10 7 colony forming units (cfu)/ml) or lipopolysaccharide (Escherichia coli J5, L-5014, Sigma, 100 ng/ml or 1000 ng/ml) for 24 hours. Tissue explants were fixed for immunohistochemistry staining of thrombin/prothrombin. Fetal membrane cells were fixed for confocal immunofluorescent staining of the biomarkers of fetal membrane cell types and thrombin/prothrombin. Protein and mRNA were harvested from the cells and tissue explants for Western blot or qRT-PCR to quantify thrombin/prothrombin protein or mRNA production, respectively. Data are presented as mean values ± standard errors of mean. Data were analyzed using one-way ANOVA with post hoc Dunnett's test. Prothrombin production and localization was confirmed by Western blot and immunostainings in all primary fetal membrane cells and tissue explants. Immunofluorescence observations revealed a perinuclear localization of prothrombin in amnion epithelial cells. Localization of prothrombin in chorion and

  3. Human gestation-associated tissues express functional cytosolic nucleic acid sensing pattern recognition receptors.

    Science.gov (United States)

    Bryant, A H; Menzies, G E; Scott, L M; Spencer-Harty, S; Davies, L B; Smith, R A; Jones, R H; Thornton, C A

    2017-07-01

    The role of viral infections in adverse pregnancy outcomes has gained interest in recent years. Innate immune pattern recognition receptors (PRRs) and their signalling pathways, that yield a cytokine output in response to pathogenic stimuli, have been postulated to link infection at the maternal-fetal interface and adverse pregnancy outcomes. The objective of this study was to investigate the expression and functional response of nucleic acid ligand responsive Toll-like receptors (TLR-3, -7, -8 and -9), and retinoic acid-inducible gene 1 (RIG-I)-like receptors [RIG-I, melanoma differentiation-associated protein 5 (MDA5) and Laboratory of Genetics and Physiology 2(LGP2)] in human term gestation-associated tissues (placenta, choriodecidua and amnion) using an explant model. Immunohistochemistry revealed that these PRRs were expressed by the term placenta, choriodecidua and amnion. A statistically significant increase in interleukin (IL)-6 and/or IL-8 production in response to specific agonists for TLR-3 (Poly(I:C); low and high molecular weight), TLR-7 (imiquimod), TLR-8 (ssRNA40) and RIG-I/MDA5 (Poly(I:C)LyoVec) was observed; there was no response to a TLR-9 (ODN21798) agonist. A hierarchical clustering approach was used to compare the response of each tissue type to the ligands studied and revealed that the placenta and choriodecidua generate a more similar IL-8 response, while the choriodecidua and amnion generate a more similar IL-6 response to nucleic acid ligands. These findings demonstrate that responsiveness via TLR-3, TLR-7, TLR-8 and RIG-1/MDA5 is a broad feature of human term gestation-associated tissues with differential responses by tissue that might underpin adverse obstetric outcomes. © 2017 British Society for Immunology.

  4. Biological wound dressings sterilized with gamma radiation: Mexican clinical experience

    Science.gov (United States)

    Martínez-Pardo, M. E.; Ley-Chávez, E.; Reyes-Frías, M. L.; Rodríguez-Ferreyra, P.; Vázquez-Maya, L.; Salazar, M. A.

    2007-11-01

    Biological wound dressings sterilized with gamma radiation, such as amnion and pig skin, are a reality in Mexico. These tissues are currently processed in the tissue bank and sterilized in the Gamma Industrial Irradiation Plant; both facilities belong to the Instituto Nacional de Investigaciones Nucleares (ININ) (National Institute of Nuclear Research). With the strong support of the International Atomic Energy Agency, the bank was established at the ININ and the Mexican Ministry of Health issued its sanitary license on July 7, 1999. The Quality Management System of the bank was certified by ISO 9001:2000 on August 1, 2003; the scope of the system is "Research, Development and Processing of Biological Tissues Sterilized with Gamma Radiation". At present, more than 150 patients from 16 hospitals have been successfully treated with these tissues. This paper presents a brief description of the tissue processing, as well as the present Mexican clinical experience with children and adult patients who underwent medical treatment with radiosterilized amnion and pig skin, used as biological wound dressings on burns and ocular surface disorders.

  5. Infections in the tissue material and their impact on the loss of transplants in the Laboratory of in vitro Cell and Tissue Culture with Tissue Bank in the years 2011-2015.

    Science.gov (United States)

    Kitala, D; Klama-Baryła, A; Kawecki, M; Kraut, M; Łabuś, W; Glik, J; Ples, M; Tomanek, E; Nowak, M

    2017-03-01

    Radiation sterilization eliminates microbiological infections but causes the degradation of the cell factor. The negative result of microbiological examination for tissue transplants is one of the conditions for approval for distribution in patients. The study attempts to verify impact of the presence of microbes onto material for transplant loss. In the 2011-2015 period, we analyzed 293 donors of skin and amnion. Microbiological sampling was performed. The total of 21 strains of bacteria, molds and fungi was identified in collected tissue. The widest spectrum of strains was found in skin (17), followed by amnia (8). The total number of positive findings was 147 and was again highest in skin (129), while the number of positive findings in amnia was 18 only. The general percentage of fungal infections was very low. The presence of fungal strains was only observed in allogeneic skin (2%). Large number of microorganisms isolated from the skin before sterilization was observed, so it seems impossible to use allogeneic intravital skin. However, the intravital application of allogeneic amnion obtained from cesarean section remains to be considered.

  6. Biological wound dressings sterilized with gamma radiation: Mexican clinical experience

    Energy Technology Data Exchange (ETDEWEB)

    Martinez-Pardo, M.E. [Instituto Nacional de Investigaciones Nucleares, Apdo. postal 18-1027, Col. Escandon 11801 Mexico DF (Mexico)], E-mail: memp@nuclear.inin.mx; Ley-Chavez, E. [ISSEMYM Toluca, Mexico DF (Mexico); Reyes-Frias, M.L. [Instituto Nacional de Investigaciones Nucleares, Apdo. postal 18-1027, Col. Escandon 11801 Mexico DF (Mexico); Rodriguez-Ferreyra, P. [Hospital ' Dr. Nicolas San Juan' , Toluca, Mexico DF (Mexico); Vazquez-Maya, L.; Salazar, M.A. [Hospital General de Mexico, Mexico DF (Mexico)

    2007-11-15

    Biological wound dressings sterilized with gamma radiation, such as amnion and pig skin, are a reality in Mexico. These tissues are currently processed in the tissue bank and sterilized in the Gamma Industrial Irradiation Plant; both facilities belong to the Instituto Nacional de Investigaciones Nucleares (ININ) (National Institute of Nuclear Research). With the strong support of the International Atomic Energy Agency, the bank was established at the ININ and the Mexican Ministry of Health issued its sanitary license on July 7, 1999. The Quality Management System of the bank was certified by ISO 9001:2000 on August 1, 2003; the scope of the system is 'Research, Development and Processing of Biological Tissues Sterilized with Gamma Radiation'. At present, more than 150 patients from 16 hospitals have been successfully treated with these tissues. This paper presents a brief description of the tissue processing, as well as the present Mexican clinical experience with children and adult patients who underwent medical treatment with radiosterilized amnion and pig skin, used as biological wound dressings on burns and ocular surface disorders.

  7. Biological wound dressings sterilized with gamma radiation: Mexican clinical experience

    International Nuclear Information System (INIS)

    Martinez-Pardo, M.E.; Ley-Chavez, E.; Reyes-Frias, M.L.; Rodriguez-Ferreyra, P.; Vazquez-Maya, L.; Salazar, M.A.

    2007-01-01

    Biological wound dressings sterilized with gamma radiation, such as amnion and pig skin, are a reality in Mexico. These tissues are currently processed in the tissue bank and sterilized in the Gamma Industrial Irradiation Plant; both facilities belong to the Instituto Nacional de Investigaciones Nucleares (ININ) (National Institute of Nuclear Research). With the strong support of the International Atomic Energy Agency, the bank was established at the ININ and the Mexican Ministry of Health issued its sanitary license on July 7, 1999. The Quality Management System of the bank was certified by ISO 9001:2000 on August 1, 2003; the scope of the system is 'Research, Development and Processing of Biological Tissues Sterilized with Gamma Radiation'. At present, more than 150 patients from 16 hospitals have been successfully treated with these tissues. This paper presents a brief description of the tissue processing, as well as the present Mexican clinical experience with children and adult patients who underwent medical treatment with radiosterilized amnion and pig skin, used as biological wound dressings on burns and ocular surface disorders

  8. Membrane Vesicles of Group B Streptococcus Disrupt Feto-Maternal Barrier Leading to Preterm Birth.

    Directory of Open Access Journals (Sweden)

    Manalee Vishnu Surve

    2016-09-01

    Full Text Available Infection of the genitourinary tract with Group B Streptococcus (GBS, an opportunistic gram positive pathogen, is associated with premature rupture of amniotic membrane and preterm birth. In this work, we demonstrate that GBS produces membrane vesicles (MVs in a serotype independent manner. These MVs are loaded with virulence factors including extracellular matrix degrading proteases and pore forming toxins. Mice chorio-decidual membranes challenged with MVs ex vivo resulted in extensive collagen degradation leading to loss of stiffness and mechanical weakening. MVs when instilled vaginally are capable of anterograde transport in mouse reproductive tract. Intra-amniotic injections of GBS MVs in mice led to upregulation of pro-inflammatory cytokines and inflammation mimicking features of chorio-amnionitis; it also led to apoptosis in the chorio-decidual tissue. Instillation of MVs in the amniotic sac also resulted in intrauterine fetal death and preterm delivery. Our findings suggest that GBS MVs can independently orchestrate events at the feto-maternal interface causing chorio-amnionitis and membrane damage leading to preterm birth or fetal death.

  9. Membrane Vesicles of Group B Streptococcus Disrupt Feto-Maternal Barrier Leading to Preterm Birth

    Science.gov (United States)

    Sthanam, Lakshmi Kavitha; Srivastava, Rohit; Basu, Bhakti; Dutta, Suryendu; Sen, Shamik; Modi, Deepak

    2016-01-01

    Infection of the genitourinary tract with Group B Streptococcus (GBS), an opportunistic gram positive pathogen, is associated with premature rupture of amniotic membrane and preterm birth. In this work, we demonstrate that GBS produces membrane vesicles (MVs) in a serotype independent manner. These MVs are loaded with virulence factors including extracellular matrix degrading proteases and pore forming toxins. Mice chorio-decidual membranes challenged with MVs ex vivo resulted in extensive collagen degradation leading to loss of stiffness and mechanical weakening. MVs when instilled vaginally are capable of anterograde transport in mouse reproductive tract. Intra-amniotic injections of GBS MVs in mice led to upregulation of pro-inflammatory cytokines and inflammation mimicking features of chorio-amnionitis; it also led to apoptosis in the chorio-decidual tissue. Instillation of MVs in the amniotic sac also resulted in intrauterine fetal death and preterm delivery. Our findings suggest that GBS MVs can independently orchestrate events at the feto-maternal interface causing chorio-amnionitis and membrane damage leading to preterm birth or fetal death. PMID:27583406

  10. Histological chorioamnionitis in preterm prelabor rupture of the membranes is associated with increased expression of galectin-3 by amniotic epithelium.

    Science.gov (United States)

    Stefanoska, Ivana; Tadić, Jasmina; Vilotić, Aleksandra; Jovanović Krivokuća, Milica; Abu Rabi, Tamara; Vićovac, Ljiljana

    2017-09-01

    Gal-3, which can regulate immune responses upon infection and inflammation, was not studied so far in intrauterine infection leading to preterm prelabor rupture of the membranes (PPROM), although gal-1 was reported to be implicated in the process. Gal-3 mRNA and protein expression in amnion and its changes during histological chorioamnionitis were studied here. Fetal membranes were obtained from women with PPROM with (n =15) and without histological chorioamnionitis (n =15) during second and third trimester. Immunohistochemical reactivity was evaluated semiquantitatively and analyzed using t-test. Galectin profile of amniotic epithelia was determined by polymerase chain reaction (PCR) and change assessed in gal-3 in PPROM with (n =5) or without histological chorioamnionitis (n =5) by real-time PCR. Human amniotic epithelium was found to express gal-1, gal-3, gal-7 and gal-8 mRNA. Gal-3 mRNA and protein is increased in fetal membranes and in the amniotic epithelium in patients with chorionamnionitis. Histological chorioamnionitis is associated with increased gal-3 expression and strong immunoreactivity of the amnion. Gal-3 may participate in the regulation of the inflammatory responses to chorioamniotic infection and/or direct interaction with pathogens.

  11. Establishment of donor Chimerism Using Allogeneic Bone Marrow with AMP Cell Co-infusion

    Science.gov (United States)

    2017-09-01

    AWARD NUMBER: W81XWH-15-1-0234 TITLE: Establishment of donor Chimerism Using Allogeneic Bone Marrow with AMP Cell Co-infusion PRINCIPAL...14/2017 4. TITLE AND SUBTITLE Establishment of donor Chimerism Using Allogeneic Bone Marrow with AMP Cell Co-infusion 5a. CONTRACT NUMBER 5b. GRANT...tolerance induction of all types of allografts. In this study, we investigate whether co-infusion of amnion- derived multipotent progenitor (AMP) cells

  12. Application of Biological Tissue Grafts for Burns in Zambia

    International Nuclear Information System (INIS)

    Chishimba, Gershom

    2001-01-01

    The author discusses the advances made in the use of Biological Tissue Grafts for the treatment of burns.The paper outlines research activities and clinical trials done in the use of gamma radiation sterilised Amnion membranes and Pig skin grafts in the zambian Heath Care System for treatment of Burns.Ethical issues of Tissue Banking are also discussed in relation to religious and cultural beliefs and Good Manufacturing Practices

  13. Experiences using IAEA Code of practice for radiation sterilization of tissue allografts: Validation and routine control

    Energy Technology Data Exchange (ETDEWEB)

    Hilmy, N. [Batan Research Tissue Bank (BRTB), Centre for Research and Development of Isotopes and Radiation Technology, P.O. Box 7002, JKSKL, Jakarta 12070 (Indonesia)], E-mail: nazly@batan.go.id; Febrida, A.; Basril, A. [Batan Research Tissue Bank (BRTB), Centre for Research and Development of Isotopes and Radiation Technology, P.O. Box 7002, JKSKL, Jakarta 12070 (Indonesia)

    2007-11-15

    Problems of tissue allografts in using International Standard (ISO) 11137 for validation of radiation sterilization dose (RSD) are limited and low numbers of uniform samples per production batch, those are products obtained from one donor. Allograft is a graft transplanted between two different individuals of the same species. The minimum number of uniform samples needed for verification dose (VD) experiment at the selected sterility assurance level (SAL) per production batch according to the IAEA Code is 20, i.e., 10 for bio-burden determination and the remaining 10 for sterilization test. Three methods of the IAEA Code have been used for validation of RSD, i.e., method A1 that is a modification of method 1 of ISO 11137:1995, method B (ISO 13409:1996), and method C (AAMI TIR 27:2001). This paper describes VD experiments using uniform products obtained from one cadaver donor, i.e., cancellous bones, demineralized bone powders and amnion grafts from one life donor. Results of the verification dose experiments show that RSD is 15.4 kGy for cancellous and demineralized bone grafts and 19.2 kGy for amnion grafts according to method A1 and 25 kGy according to methods B and C.

  14. RESEARCH ON REDUCING PREMATURITY RUPTURE OF MEMBRANE

    OpenAIRE

    Maria URSACHI (BOLOTA); Emil ANTON; Sorana Caterina ANTON

    2016-01-01

    The membranes surrounding the amniotic cavity are composed from amnion and chorion, tightly adherent layers which are composed of several cell types, including epithelial cells, trophoblasts cells and mesenchyme cells, embedded in a collagenous matrix. They retain amniotic fluid, secret substances into the amniotic fluid, as well as to the uterus and protect the fetus against upward infections from urogenital tract. Normally, the membranes it breaks during labor. Premature rupture of the amn...

  15. The impact of the International Atomic Energy Agency (IAEA) program on radiation and tissue banking in Mexico.

    Science.gov (United States)

    Martínez-Pardo, María Esther; Morales Pedraza, Jorge; Sánchez Ramírez, Omar

    2009-05-01

    Tissue banking started in Mexico in 1948-1949, when two bone banks were established, one at the Infantile Hospital of Mexico and other at the Central Military Hospital. Mexico has benefited for the implementation of the IAEA program since through it has been able to settle down and to consolidate the Tissue Bank at the Instituto Nacional de Investigaciones Nucleares ININ (National Institute for Nuclear Research). This is the only bank in Latin America that has a Quality Management System in force, certified under ISO 9001:2000 since August 1, 2003. The first tissue processed was amnion. The main products of the BTR are amnion and pig skin. Both are biological tissues which their main use is as a wound dressing in patients with burns, scars, diabetic ulcers, epidermolysis bullosa, damaged ocular surface, etc. The General Health Law, published in 1984 and reformed in June 19, 2007, describes the procedure for the disposal of organs, tissues and human cadavers in its fourteenth title and in the Regulation for Sanitary Control. During the period 2001-2005, the ININ Tissue Bank produced 292 sterilised tissues (amnion, 86,668 cm(2), and frozen pig skin, 164,220 cm(2), at an estimated cost of 1,012,668 Mexican pesos. Until 2006, one hundred eighty five (185) patients have been treated with the use of sterilised tissues produced by the ININ Tissue Bank. The radiation source used for sterilisation of tissues is an industrial Cobalt-60 irradiator model JS-6500 AECL, which belongs to ININ. This equipment is located in other building, close to the BTR, in the Centro Nuclear de México "Dr. Nabor Carrillo Flores" (Nuclear Center of Mexico). Until 2006, six hospitals use in a routine way the sterilised tissues produced by the ININ Tissue Bank, for the treatment of burns originated by diverse agents like flame, electricity, liquids in boil, chemical reagents, as well as for the reconstruction of the ocular surface. Two of these hospitals treat patients of very low economic

  16. Radiosterilization of freeze-dried human amniotic Membrane and its use in the treatment of burn wound. Algerian experience

    International Nuclear Information System (INIS)

    Djefal, A.; Mahlous, M.; Nacer Khodja, A.; Larbi, M.; Larbi Daho Bachir, M.

    2001-01-01

    The present study evaluates the usefulness of human amniotic membrane as biological dressing and its efficacy in the treatment of burns comparatively to the conventional dressing. We reported the practical methods of preparation, preservation and radiation sterilisation of amnion, and the clinical results of its successful use in the treatment of 80 cases of superficial and intermediate depth dermal burns. The increased rate of healing, pain relief, good adhesion to the bed wound and absence of infection were observed

  17. Myocytes of chorionic vessels from placentas with meconium-associated vascular necrosis exhibit apoptotic markers.

    Science.gov (United States)

    King, Erin L; Redline, Raymond W; Smith, Steven D; Kraus, Frederick T; Sadovsky, Yoel; Nelson, D Michael

    2004-04-01

    Meconium-associated vascular necrosis (MAVN) is a histological abnormality of human placental chorionic vessels that is associated with poor neonatal outcome. We tested the hypothesis that MAVN shows apoptosis in the walls of chorionic vessels. Archival placental specimens with MAVN (n = 5) were compared with specimens from uncomplicated pregnancies at term (n = 5) and from placentas with intense chorionic vasculitis associated with acute chorioamnionitis with (n = 5) or without (n = 5) a clinical history of meconium in the amniotic fluid. Sections from all placentas were processed by the TUNEL method, and 2 observers who were blinded to specimen diagnosis quantified the immunofluorescent TUNEL staining in both the amnion-facing and villous-facing walls of the larger chorionic vessels in each specimen. Compared with the other 3 groups, only the amnion-facing wall of chorionic vessels in MAVN showed a significantly greater number of apoptotic cells. This was verified by morphological criteria and caspase 3 staining. There were limited or no detectable TUNEL-stained cells in either the villous-facing walls of vessels in the MAVN specimens or in any of the vessels of the placentas from uncomplicated pregnancies. There was a negligible level of apoptosis in chorionic vessels of placentas with intense chorionic vasculitis, with or without meconium, despite the inflammatory response or presence of meconium. We conclude that apoptosis contributes to the pathophysiology of MAVN.

  18. Primordial germ cells and amnion development in the avian embryo

    NARCIS (Netherlands)

    De Melo Bernardo, Ana

    2016-01-01

    Primordial germ cells (PGCs) are the progenitors of the gametes, responsible for transmitting genetic information from generation to generation. Although there is a long history of gamete biology research, there is still a lot to be learned about many of the mechanisms underlying germ cell

  19. Amniotic band syndrome: A clinical brief

    Directory of Open Access Journals (Sweden)

    Dasaradha Ramireddy Malireddy

    2017-01-01

    Full Text Available Amniotic band syndrome (ABS results from bands of amnion entangling fetal parts. They may manifest as constriction rings or complex congenital anomalies resulting in stillbirth. Karyotyping is important for exclusion of inherited disorders and proper counseling. Two case reports one stillbirth and the other with constriction ring of fingers and mild hydronephrosis are presented. The aim of this paper is to make awareness and stress the need for doing thorough work-up in all cases of constriction bands.

  20. Human Decidua-Derived Mesenchymal Cells Are a Promising Source for the Generation and Cell Banking of Human Induced Pluripotent Stem Cells

    OpenAIRE

    Shofuda, Tomoko; Kanematsu, Daisuke; Fukusumi, Hayato; Yamamoto, Atsuyo; Bamba, Yohei; Yoshitatsu, Sumiko; Suemizu, Hiroshi; Nakamura, Masato; Sugimoto, Yoshikazu; Furue, Miho Kusuda; Kohara, Arihiro; Akamatsu, Wado; Okada, Yohei; Okano, Hideyuki; Yamasaki, Mami

    2012-01-01

    Placental tissue is a biomaterial with remarkable potential for use in regenerative medicine. It has a three-layer structure derived from the fetus (amnion and chorion) and the mother (decidua), and it contains huge numbers of cells. Moreover, placental tissue can be collected without any physical danger to the donor and can be matched with a variety of HLA types. The decidua-derived mesenchymal cells (DMCs) are highly proliferative fibroblast-like cells that express a similar pattern of CD a...

  1. Uterine overdistention induces preterm labor mediated by inflammation: observations in pregnant women and nonhuman primates

    Science.gov (United States)

    Waldorf, Kristina M. Adams; Singh, Natasha; Mohan, Aarthi R.; Young, Roger C.; Ngo, Lisa; Das, Ananya; Tsai, Jesse; Bansal, Aasthaa; Paolella, Louis; Herbert, Bronwen R.; Sooranna, Suren R.; Gough, G. Michael; Astley, Cliff; Vogel, Keith; Baldessari, Audrey E.; Bammler, Theodor K.; MacDonald, James; Gravett, Michael G.; Rajagopal, Lakshmi; Johnson, Mark R.

    2015-01-01

    OBJECTIVE Uterine overdistention is thought to induce preterm labor in women with twin and multiple pregnancies, but the pathophysiology remains unclear. We investigated for the first time the pathogenesis of preterm birth associated with rapid uterine distention in a pregnant nonhuman primate model. STUDY DESIGN A nonhuman primate model of uterine overdistention was created using preterm chronically catheterized pregnant pigtail macaques (Macaca nemestrina) by inflation of intraamniotic balloons (N = 6), which were compared to saline controls (N = 5). Cesarean delivery was performed due to preterm labor or at experimental end. Microarray, quantitative reverse transcriptase polymerase chain reaction, Luminex (Austin, TX), and enzyme-linked immunosorbent assay were used to measure messenger RNA (mRNA) and/or protein levels from monkey (amniotic fluid, myometrium, maternal plasma) and human (amniocytes, amnion, myometrium) tissues. Statistical analysis employed analysis of covariance and Wilcoxon rank sum. Biomechanical forces were calculated using the law of Laplace. RESULTS Preterm labor occurred in 3 of 6 animals after balloon inflation and correlated with greater balloon volume and uterine wall stress. Significant elevations of inflammatory cytokines and prostaglandins occurred following uterine overdistention in an “inflammatory pulse” that correlated with preterm labor (interleukin [IL]-1β, tumor necrosis factor [TNF]-α, IL-6, IL-8, CCL2, prostaglandin E2, prostaglandin F2α, all P < .05). A similar inflammatory response was observed in amniocytes in vitro following mechanical stretch (IL1β, IL6, and IL8 mRNA multiple time points, P < .05), in amnion of women with polyhydramnios (IL6 and TNF mRNA, P < .05) and in amnion (TNF-α) and myometrium of women with twins in early labor (IL6, IL8, CCL2, all P < .05). Genes differentially expressed in the nonhuman primate after balloon inflation and in women with polyhydramnios and twins are involved in tissue

  2. Expression and function of NOD-like receptors by human term gestation-associated tissues.

    Science.gov (United States)

    Bryant, Aled H; Bevan, Ryan J; Spencer-Harty, Samantha; Scott, Louis M; Jones, Ruth H; Thornton, Catherine A

    2017-10-01

    Nucleotide-binding oligomerization domain (NOD)-like receptors or NOD-like receptors (NLRs) have been implicated in several disease pathologies associated with inflammation. Since local and systemic inflammation is a hallmark of both term and preterm labour, a role for NLRs at the materno-fetal interface has been postulated. Gene expression and immunolocalisation of NLR family members in human placenta, choriodecidua, and amnion were examined. Tissue explants were used to examine the response to activators of NOD1 (Tri-DAP), NOD2 (MDP) and NLRP3 (nigericin). Cell/tissue-free supernatants were examined for the production of interleukin (IL)-1β, IL-6, IL-8 and IL-10 using specific ELISAs. Expression of transcripts for NOD1, NOD2, NLRP3, NLRC4, NLRX1, NLRP1 and NAIP and protein expression of NOD1, NOD2 and NLRP3 were a broad feature of all term gestation-associated tissues. Production of cytokines was increased significantly in response to all ligands in placenta and choriodecidua, except for MDP-induced IL-10. Similarly, there was a significant in the amnion except for MDP induced IL-1β and IL-10 response to either agonist. IL-1β production was dependent on caspase-1 regardless of agonist used or tissue examined. Term human gestation-associated tissues express functional NLRs which likely play a role in both sterile and pathogen-driven inflammatory responses at the materno-fetal interface. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Dietary Flavonoids as Therapeutics for Preterm Birth: Luteolin and Kaempferol Suppress Inflammation in Human Gestational Tissues In Vitro

    Science.gov (United States)

    Wall, Courtney; Lim, Ratana; Poljak, Marin; Lappas, Martha

    2013-01-01

    Infection/inflammation is commonly associated with preterm birth (PTB), initiating uterine contractions and rupture of fetal membranes. Proinflammatory cytokines induce matrix metalloproteinases (MMPs) that degrade the extracellular matrix (ECM) and prostaglandins which initiate uterine contractions. Nuclear factor-κB (NF-κB) and activator-protein- (AP-)1 have key roles in the formation of these prolabour mediators. In nongestational tissues, dietary flavonoids such as luteolin and kaempferol inhibit NF-κB, AP-1, and their downstream targets. The aim of this study was to determine if luteolin and kaempferol reduce infection-induced prolabour mediators in human gestational tissues. Fetal membranes were incubated with LPS, and primary amnion cells and myometrial cells were incubated with IL-1β in the absence or presence of luteolin or kaempferol. Luteolin and kaempferol significantly reduced LPS-induced secretion of proinflammatory cytokines (IL-6 and IL-8) and prostaglandins (PGE2 and PGF2α) in fetal membranes, IL-1β-induced COX-2 gene expression and prostaglandin production in myometrium, and IL-1β-induced MMP-9 activity in amnion and myometrial cells. Luteolin and kaempferol decreased IL-1β-induced NF-κB p65 DNA binding activity and nuclear c-Jun expression. In conclusion, luteolin and kaempferol inhibit prolabour mediators in human gestational tissues. Given the central role of inflammation in provoking preterm labour, phytophenols may be a therapeutic approach to reduce the incidence of PTB. PMID:23840918

  4. Amniotic membrane for burn trauma

    International Nuclear Information System (INIS)

    Jamaluddin Zainol; Hasim Mohammad

    1999-01-01

    Amniotic membranes are derived from human placentae at birth. They have two layers mainly the amniotic and the chorionic surfaces which are separated by a thin layer of connective tissues. The two layers are separated during procurement, the placenta and the chorionic side are discarded and the amnion membranes are then further processed. Amnion membranes are normally procured from placentae which are normally free of infections, i.e; the mothers are antenatally screened for sexually transmitted diseases or AlDs related diseases. Intrapartum the mother should not be having chorioamnionitis or jaundice. Sometimes the amniotic membranes are acquired from fresh elective caeserian sections. After processing, the amniotic membranes are packed in two layers of polypropylene and radiated with cobalt 60 at a dose of about 25 kGy. The amniotic membranes are clinically used to cover burn surfaces especially effective for superficial or partial thickness burns. The thin membranes adhered well to the trauma areas and peeled off automatically by the second week. No change of dressing were necessary during these times because of the close adherence, there were less chance of external contamination or infections of these wounds. Due to their flexibility they are very useful to cover difference contours of the human body for example the face, body, elbows or knees. However our experience revealed that amniotic membranes are not useful for third degree bums because the membranes dissolves by the enzymes present in the wounds

  5. Stemcell Information: SKIP000652 [SKIP Stemcell Database[Archive

    Lifescience Database Archive (English)

    Full Text Available both neonatal and maternal cells. 羊膜(母胎・胎児)由来線維芽細胞(HFM-1)由来iPS細胞. human ES-like Research Grade Retrovirus Oc... SKIP000652 ... amnion 羊膜 Normal HiPS-RIKEN-3D HiPS-RIKEN-3D ... Fetus Male ... -- No Human iPS cel...l line. Parent cell line of HFM-1(amniotic membrane cells).HFM-1 were derived from ...2010.00091.x Establishment of induced pluripotent stem cells from human neonatal tissues. Fujioka T, Shimizu

  6. Stemcell Information: SKIP000655 [SKIP Stemcell Database[Archive

    Lifescience Database Archive (English)

    Full Text Available both neonatal and maternal cells. 羊膜(母胎・胎児)由来線維芽細胞(HFM-1)由来iPS細胞. human ES-like Research Grade Lentivirus Oc... SKIP000655 ... amnion 羊膜 Normal HiPS-RIKEN-4B HiPS-RIKEN-4B ... Fetus Male ... -- No Human iPS cel...l line. Parent cell line of HFM-1(amniotic membrane cells).HFM-1 were derived from ...4.2010.00091.x Establishment of induced pluripotent stem cells from human neonatal tissues. Fujioka T, Shimi

  7. The MRC-5 human embryonal lung fibroblast two-dimensional gel cellular protein database: quantitative identification of polypeptides whose relative abundance differs between quiescent, proliferating and SV40 transformed cells

    DEFF Research Database (Denmark)

    Celis, J E; Dejgaard, K; Madsen, Peder

    1990-01-01

    interferon-induced proteins, were not detected in the master MRC-5 images. The identity of 36 of the transformation-sensitive proteins whose levels are up or down regulated by two times or more was determined and additional information can be transferred from the master transformed human epithelial amnion......, this comprehensive database will outline an integrated picture of the expression levels and properties of the thousands of protein components of organelles, pathways and cytoskeletal systems that may be directly or indirectly involved in properties associated with the transformed state. Udgivelsesdato: 1990-Dec...

  8. Stemcell Information: SKIP000653 [SKIP Stemcell Database[Archive

    Lifescience Database Archive (English)

    Full Text Available both neonatal and maternal cells. 羊膜(母胎・胎児)由来線維芽細胞(HFM-1)由来iPS細胞. human ES-like Research Grade Retrovirus Oc... SKIP000653 ... amnion 羊膜 Normal HiPS-RIKEN-3E HiPS-RIKEN-3E ... Fetus Male ... -- No Human iPS cel...l line. Parent cell line of HFM-1(amniotic membrane cells).HFM-1 were derived from ...2010.00091.x Establishment of induced pluripotent stem cells from human neonatal tissues. Fujioka T, Shimizu

  9. Evaluation of Amnion-derived Multipotent Progenitor (AMP) Cells and Amnion-derived Cellular Cytokine Solution (ST266) in Promoting Craniomaxillofacial Regenerative Bone Healing in Critical Size Calvarial Defects

    Science.gov (United States)

    2017-10-10

    well as diseases that can lead to eventual bone loss such as periodontitis (currently being evaluated in human clinical trials). A previous report...28 using ST266 treatment in a rabbit periodontal model revealed that ST266 treatment reduced inflammation and had increased new bone formation [32...The increase in new bone formation may have arisen due to the reduction in chronic inflammation by ST266 treatment, thus allowing the initiation

  10. Hepatitis A virus cellular receptor 2 (HAVCR2) is decreased with viral infection and regulates pro-labour mediators OA.

    Science.gov (United States)

    Liong, Stella; Lim, Ratana; Barker, Gillian; Lappas, Martha

    2017-07-01

    Intrauterine infection caused by viral infection has been implicated to contribute to preterm birth. Hepatitis A virus cellular receptor 2 (HAVCR2) regulates inflammation in non-gestational tissues in response to viral infection. The aims of this study were to determine the effect of: (i) viral dsRNA analogue polyinosinic:polycytidylic acid (poly(I:C)) on HAVCR2 expression; and (ii) HAVCR2 silencing by siRNA (siHAVCR2) in primary amnion and myometrial cells on poly(I:C)-induced inflammation. In human foetal membranes and myometrium, HAVCR2 mRNA and protein expression was decreased when exposed to poly(I:C). Treatment of primary amnion and myometrial cells with poly(I:C) significantly increased the expression and release of pro-inflammatory cytokines TNF, IL1A, IL1B and IL6; the expression of chemokines CXCL8 and CCL2; the expression and secretion of adhesion molecules ICAM1 and VCAM1; and PTGS2 and PTGFR mRNA expression and the release of prostaglandin PGF 2α . This increase was significantly augmented in cells transfected with siHAVCR2. Furthermore, mRNA expression of anti-inflammatory cytokines IL4 and IL10 was significantly decreased. Collectively, our data suggest that HAVCR2 regulates cytokines, chemokines, prostaglandins and cell adhesion molecules in the presence of viral infection. This suggests a potential for HAVCR2 activators as therapeutics for the management of preterm birth associated with viral infections. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. High plasticity in epithelial morphogenesis during insect dorsal closure

    Directory of Open Access Journals (Sweden)

    Kristen A. Panfilio

    2013-09-01

    Insect embryos complete the outer form of the body via dorsal closure (DC of the epidermal flanks, replacing the transient extraembryonic (EE tissue. Cell shape changes and morphogenetic behavior are well characterized for DC in Drosophila, but these data represent a single species with a secondarily reduced EE component (the amnioserosa that is not representative across the insects. Here, we examine DC in the red flour beetle, Tribolium castaneum, providing the first detailed, functional analysis of DC in an insect with complete EE tissues (distinct amnion and serosa. Surprisingly, we find that differences between Drosophila and Tribolium DC are not restricted to the EE tissue, but also encompass the dorsal epidermis, which differs in cellular architecture and method of final closure (zippering. We then experimentally manipulated EE tissue complement via RNAi for Tc-zen1, allowing us to eliminate the serosa and still examine viable DC in a system with a single EE tissue (the amnion. We find that the EE domain is particularly plastic in morphogenetic behavior and tissue structure. In contrast, embryonic features and overall kinetics are robust to Tc-zen1RNAi manipulation in Tribolium and conserved with a more distantly related insect, but remain substantially different from Drosophila. Although correct DC is essential, plasticity and regulative, compensatory capacity have permitted DC to evolve within the insects. Thus, DC does not represent a strong developmental constraint on the nature of EE development, a property that may have contributed to the reduction of the EE component in the fly lineage.

  12. Optimal treatment of partial thickness burns in children: a systematic review.

    Science.gov (United States)

    Vloemans, A F P M; Hermans, M H E; van der Wal, M B A; Liebregts, J; Middelkoop, E

    2014-03-01

    A large part of the patient population of a burn centre consists of children, most of whom are younger than four years. The majority of these young children suffer from superficial and deep partial thickness scald burns that may easily deepen to full thickness burns. A proper wound therapy, that prevents infection and ensures a moist wound condition, might prevent the deterioration of the wound. Therefore, we performed a systematic review of wound management and dressing materials to select the best treatment option for children with burns. A search in Medline and Embase revealed 51 articles for a critical appraisal. The articles were divided into randomized controlled trials, cohort studies and a group of case-reports. Total appraisal did not differ much amongst the groups; the level of evidence was highest in the randomized controlled trials and lowest in the case-reports. In 16 out of 34 comparative studies, silver sulfadiazine or a silver sulfadiazine/chlorhexidine-gluconate combination was the standard of wound care treatment. The competitor dressing was Biobrane(®) in six studies and amnion membrane in three. Tulle gauze, or tulle gauze impregnated with an antibacterial addition were the standard of care treatment in seven studies. In general, membranous dressings like Biobrane(®) and amnion membrane performed better than the standard of care on epithelialization rate, length of hospital stay and pain for treatment of partial thickness burns in children. However, hardly any of the studies investigated long-term results like scar formation. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

  13. The tissue bank at the national nuclear research institute in Mexico.

    Science.gov (United States)

    Esther Martínez-Pardo, María; Lourdes Reyes-Frías, Ma

    2003-01-01

    The Instituto Nacional de Investigaciones Nucleares (ININ, The National Nuclear Research Institute) received during 1997-1998 strong support of the International Atomic Energy Agency (IAEA), to establish the first and only one tissue bank (BTR ININ tissue bank) in Mexico that uses ionising radiation as sterilising agent. In that time, the BTR staff was trained in different tissue banks in several countries. Basic equipment for tissue processing donated by the IAEA was received in 1998. In July, 1999 the Mexican Health Secretariat gave the Sanitary License No. 1062000001 to the BTR to operate as an official organ and tissue bank. In August, 2001 the ININ and the Hospital Materno Infantil (HMI-ISSEMYM) signed an agreement to collaborate in amnion processing. The hospital is responsible for donor selection, serology tests, tissue procurement and washing, since this hospital is the BTR amnion supplier. The tissues are collected by ININ weekly with complete documentation. The BTR is responsible for processing: cleaning, air drying, packaging, labelling, microbiological control and sterilisation by gamma irradiation. The sterilised tissue is kept under quarantine for 6 months to obtain the results of the donor second serology test. From March to June, 2002 the BTR has processed 347.86 units (50 cm(2) each), is say, 17,393 cm(2). In addition, the pig skin xenograft process has been implemented and a protocol for clinical applications of it is running at the Hospital Central Sur de Alta Especialidad (PEMEX). Also the ININ tissue bank present status and perspectives are described.

  14. Activation of AMPK in human fetal membranes alleviates infection-induced expression of pro-inflammatory and pro-labour mediators.

    Science.gov (United States)

    Lim, R; Barker, G; Lappas, M

    2015-04-01

    In non-gestational tissues, the activation of adenosine monophosphate (AMP)-activated kinase (AMPK) is associated with potent anti-inflammatory actions. Infection and/or inflammation, by stimulating pro-inflammatory cytokines and matrix metalloproteinase (MMP)-9, play a central role in the rupture of fetal membranes. However, no studies have examined the role of AMPK in human labour. Fetal membranes, from term and preterm, were obtained from non-labouring and labouring women, and after preterm pre-labour rupture of membranes (PPROM). AMPK activity was assessed by Western blotting of phosphorylated AMPK expression. To determine the effect of AMPK activators on pro-inflammatory cytokines, fetal membranes were pre-treated with AMPK activators then stimulated with bacterial products LPS and flagellin or viral dsDNA analogue poly(I:C). Primary amnion cells were used to determine the effect of AMPK activators on IL-1β-stimulated MMP-9 expression. AMPK activity was decreased with term labour. There was no effect of preterm labour. AMPK activity was also decreased in preterm fetal membranes, in the absence of labour, with PROM compared to intact membranes. AMPK activators AICAR, phenformin and A769662 significantly decreased IL-6 and IL-8 stimulated by LPS, flagellin and poly(I:C). Primary amnion cells treated with AMPK activators significantly decreased IL-1β-induced MMP-9 expression. The decrease in AMPK activity in fetal membranes after spontaneous term labour and PPROM indicates an anti-inflammatory role for AMPK in human labour and delivery. The use of AMPK activators as possible therapeutics for threatened preterm labour would be an exciting future avenue of research. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Ultrasonographic diagnosis of limb-body wall complex: a case report

    Energy Technology Data Exchange (ETDEWEB)

    Goo, Dong Erk; Yoo, Shi Joon; Cho, Kyoung Sik; Choi, Kee Young; Kim, Am [University of Ulsan College of Medicine, Asan Medical Center, Ulsan (Korea, Republic of)

    1993-12-15

    Limb-body wall complex (LBWC) is a rare congenital process believed to result from early rupture of the amnion, possibly as a consequence of vascular disruption. Characteristic fetal deformities include limb defects, scoliosis, thoracoschisis and/or abdominoschisis, and severe craniofacial anomalies. This complex should be distinguished from other body-wall defect including omphalocele and gastroschisis since the prognosis of LBWC is uniformly poor. We report a case of LBWC which was diagnosed by prenatal ultrasonography in 26 weeks of gestation. Prenatal ultrasonograms are correlated with the gross pathologic findings. The present case demonstrates the superb capability of modern ultrasonography in the diagnosis of complex fetal anomalies

  16. Ultrasonographic diagnosis of limb-body wall complex: a case report

    International Nuclear Information System (INIS)

    Goo, Dong Erk; Yoo, Shi Joon; Cho, Kyoung Sik; Choi, Kee Young; Kim, Am

    1993-01-01

    Limb-body wall complex (LBWC) is a rare congenital process believed to result from early rupture of the amnion, possibly as a consequence of vascular disruption. Characteristic fetal deformities include limb defects, scoliosis, thoracoschisis and/or abdominoschisis, and severe craniofacial anomalies. This complex should be distinguished from other body-wall defect including omphalocele and gastroschisis since the prognosis of LBWC is uniformly poor. We report a case of LBWC which was diagnosed by prenatal ultrasonography in 26 weeks of gestation. Prenatal ultrasonograms are correlated with the gross pathologic findings. The present case demonstrates the superb capability of modern ultrasonography in the diagnosis of complex fetal anomalies

  17. The impact of the International Atomic Energy Agency (IAEA) program on radiation and tissue banking in Peru.

    Science.gov (United States)

    Gamero, Emma Castro; Morales Pedraza, Jorge

    2009-05-01

    The tissue bank "Rosa Guerzoni Chambergo" (RGCTB) located at the Child's Health Institute was inaugurated in 1996, with the financial and technical support of the IAEA program on radiation and tissue banking. Since 1998, the biological bandage of fresh and lyophilised pigskin, amnion and bone tissue is processed routinely in this bank. In all cases, the tissue is sterilised with the use of Cobalt-60 radiation, process carried out at the Laboratories of Irradiation of the Peruvian Institute of Nuclear Energy (IPEN). The tissue bank in the Child's Health Institute helped to save lives in an accident occurred in Lima, when a New Year's fireworks celebration ran out of control in January 2002. Nearly 300 people died in the tragic blaze and hundreds more were seriously burned and injured. Eight Lima hospitals and clinics suddenly were faced with saving the lives of severely burned men, women and children. Fortunately, authorities were ready to respond to the emergency. More than 1,600 dressings were sterilised and supplied to Lima surgeons. The efforts helped save the lives of patients who otherwise might not have survived the Lima fire. Between 1998 and September 2007, 35,012 tissue grafts were produced and irradiated. Radiation sterilised tissues are used by 20 national medical institutions as well as 17 private health institutions. The tissue bank established in Peru with the support of the IAEA is now producing the following tissues: pigskin dressings, fresh and freeze-dried; bone allografts, chips, wedges and powdered, and amnion dressings air-dried. It is also now leading the elaboration of national standards, assignment being entrusted by ONDT (Organización Nacional de Donación y Transplantes; National Organisation on Donation and Transplant). This among other will permit the accreditation of the tissue bank. In this task is also participating IPEN.

  18. Phenotypic Screening Identifies Synergistically Acting Natural Product Enhancing the Performance of Biomaterial Based Wound Healing

    Directory of Open Access Journals (Sweden)

    Srinivasan Sivasubramanian

    2017-07-01

    Full Text Available The potential of multifunctional wound heal biomaterial relies on the optimal content of therapeutic constituents as well as the desirable physical, chemical, and biological properties to accelerate the healing process. Formulating biomaterials such as amnion or collagen based scaffolds with natural products offer an affordable strategy to develop dressing material with high efficiency in healing wounds. Using image based phenotyping and quantification, we screened natural product derived bioactive compounds for modulators of types I and III collagen production from human foreskin derived fibroblast cells. The identified hit was then formulated with amnion to develop a biomaterial, and its biophysical properties, in vitro and in vivo effects were characterized. In addition, we performed functional profiling analyses by PCR array to understand the effect of individual components of these materials on various genes such as inflammatory mediators including chemokines and cytokines, growth factors, fibroblast stimulating markers for collagen secretion, matrix metalloproteinases, etc., associated with wound healing. FACS based cell cycle analyses were carried out to evaluate the potential of biomaterials for induction of proliferation of fibroblasts. Western blot analyses was done to examine the effect of biomaterial on collagen synthesis by cells and compared to cells grown in the presence of growth factors. This work demonstrated an uncomplicated way of identifying components that synergistically promote healing. Besides, we demonstrated that modulating local wound environment using biomaterials with bioactive compounds could enhance healing. This study finds that the developed biomaterials offer immense scope for healing wounds by means of their skin regenerative features such as anti-inflammatory, fibroblast stimulation for collagen secretion as well as inhibition of enzymes and markers impeding the healing, hydrodynamic properties complemented

  19. Comparative evaluation of coronally advanced flap using amniotic membrane and platelet-rich fibrin membrane in gingival recession: An 18-month clinical study

    Directory of Open Access Journals (Sweden)

    Mohd Rehan

    2018-01-01

    Full Text Available Background: An amnion membrane is a placenta-derived tissue that consists of numerous growth factors, proteins, and stem cell reserves which help in accelerated wound healing and regeneration. Platelet-rich fibrin (PRF also releases growth factors after activation from the platelets and gets trapped within fibrin matrix which has been shown to stimulate the mitogenic response in the periosteum for bone repair and regeneration during normal wound healing. This preliminary, controlled, randomized clinical trial with an 18-month follow-up was aimed to evaluate the effectiveness of coronally advanced flap (CAF with either PRF membrane or bioresorbable amniotic membrane (AM in treatment of localized gingival recession defects. Materials and Methods: Sixteen healthy adult patients presenting with Miller Class I recession defects were treated surgically with CAF along with AM (Group I or PRF (Group II for coverage of the recession defects. For all patients, plaque index, gingival index, bleeding on probing, clinical attachment level, depth of recession, width of recession, width of attached gingiva, and gingival thickness were evaluated at 6 months and 18 months postoperatively. Statistical analysis was done using paired t-test, repeated measure analysis of variance test, Bonferroni test for intragroup comparison and unpaired t-test for intergroup comparison. Results: The results showed statistically nonsignificant (P < 0.01 difference in all clinical parameters at the 6- and 18-month follow-ups in both groups. Gingival recession in both PRF and amnion group when evaluated individually, significantly reduced from baseline to 6 months (P = 0.000 and from baseline to 18 months (P = 0.000. However, the mean value from 6 months to 18 months was statistically nonsignificant. Conclusion: The present study demonstrated that both CAF + PRF and CAF + AM are equally effective in providing clinically significant outcomes with respect to root coverage with AM

  20. [HIV and pregnancy: 2013 guidelines from the French expert working group].

    Science.gov (United States)

    Mandelbrot, L; Berrébi, A; Matheron, S; Blanche, S; Tubiana, R; Rouzioux, C; Faucher, P; Partisani, M; Boyer, V; Taeron, C; Faye, A; Bujan, L; Dabis, F; Warszawski, J; Morlat, P

    2014-09-01

    With effective antiretroviral therapy, the risk of mother to child transmission (MTCT) is now under 1%. The 2013 French guidelines emphasize early antiretroviral lifelong antiretroviral therapy. Thus, the current trend for women living with HIV is to take antiretroviral therapy before, during and after their pregnancies. A major issue today is the choice of antiretroviral drugs, to maximize the benefits and minimize the risks of fetal exposure. This requires interdisciplinary care. The use of effective therapies permits gradual but profound changes in obstetric practice. When maternal plasma viral load is controlled (pregnancy is above 400 copies/mL. Intravenous zidovudine during labor is recommended only if the last maternal viral load is>400 copies/mL or in case of complications such as preterm delivery, bleeding or chorio-amnionitis during labor. In case of premature rupture of membranes before 34 weeks, a multidisciplinary decision should be made, based on gestational age and control of maternal viral load; if the woman is under antiretroviral therapy and especially if her viral load is undetectable, steroids and antibiotics should be offered and pregnancy can be continued except in case of signs or symptoms of chorio-amnionitis. Breastfeeding is not recommended in women living with HIV in France, as in industrialized countries. Prophylaxis in the newborn is usually zidovudine for 1 month. In case of significant exposure to HIV perinatally, in particular when, maternal viral load is>1000 copies/mL, prophylactic combination therapy is recommended. Monitoring of the child is necessary to determine whether or not it is free of HIV infection and to monitor possible adverse effects of perinatal exposure to antiretroviral drugs. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  1. Bronchial stump closure with amniotic membrane in animal model

    Directory of Open Access Journals (Sweden)

    Gholamreza Mohajeri

    2014-01-01

    Full Text Available Background: Coverage of the bronchial stumps (BSs with adjacent tissues can improve healing and reduce bronchial complications in complex thoracic surgery. There is no evidence for the application of human amnion allograft for prevention of air leak from the BS. The comparison of the amniotic membrane (AM and pleural patch for BS healing after lobectomy in dogs was our aim in this study. Materials and Methods: A total of eight males and females 12-24-month-old dogs between 17 and 22 kg body-weight were used in this study in 2010, Isfahan University of Medical Sciences. Animals were separated into two groups: group A (n = 4; amniotic membrane and group P (n = 4; pleural patch according to the BS closure technique performed. After lobectomy of the right middle lobe, the BS was closed, while a small bronchopleural fistula (BPF was created by inserting a catheter via edges of closed stump. Then, it was covered with a piece of AM3 × 3 cm in group A and with a pedicle graft of pleura in group P. Rethoracotomy was performed after 15 days of observation, and the BS was removed for histological examination. Histological healing was classified as complete or incomplete healing. Neoangiogenesis was measured by Von Willebrand expression using immunohistochemistry (IHC. Data were analyzed by SPSS version 15 using Fisher′s exact test, Mann-Whitney test, and T tests. Results: BPF complications were not seen during observation period. There was no significant difference in histological healing between two groups. Similarly, no significant difference was observed between the groups in terms of neoangiogenesis based on IHC examination (P value = 0.69. Conclusion: Human amnion allograft could be as effective as pleural patch for BS wrapping following pulmonary resections.

  2. Production of reference enteroviruses.

    Science.gov (United States)

    Kalter, S S; Rodriguez, A R; Armour, V

    1968-10-01

    Forty-five human enterovirus reagents of certified purity and quality were prepared for use as seed viruses and as immunizing antigens. One of the reagents was ampouled as "untreated" seed virus, whereas 14 were ampouled as "MgCl(2)-stabilized" reagents. The remaining 30 reagents were ampouled as "untreated" seed viruses and as "MgCl(2)-stabilized" reagents. Thirty of the reagents were propagated on primary African green monkey kidney cells, 3 on primary baboon kidney cells, 3 on primary rhesus monkey kidney cells, and the remaining 9 on human amnion cells. Forty-two of the viral antigens were concentrated for use in the production of high-titered specific antisera in large animals.

  3. Ionization radiation in sterilization of the tissue transplants

    International Nuclear Information System (INIS)

    Uhrynowska-Tyszkiewicz, I.; Kaminski, A.

    2007-01-01

    Established in 1963, the Central Tissue Bank in Warsaw is a multi-tissue bank located in the Department of Transplantology of the Medical University in Warsaw. Allografts such as bone, cartilage, tendons, ligaments, sclera, skin and amnion are preserved mainly by deep-freezing and/or lyophilization and subsequently radiation-sterilized with a dose of 35 kGy with gamma rays in a 60 Co source (at the Institute of Applied Radiation Chemistry in Lodz) or with electron beam 10 MeV accelerator (at the Institute of Nuclear Chemistry and Technology in Warsaw). This is the oldest working tissue bank in the world, which for almost 40 years now has routinely used ionizing radiation for sterilization of tissue allografts

  4. The fine structure of human germ layers in vivo: clues to the early differentiation of embryonic stem cells in vitro.

    Science.gov (United States)

    Sathananthan, Henry; Selvaraj, Kamala; Clark, Joan

    2011-08-01

    The fine structure of the three germ layers in human ectopic embryos (stage 7) have been documented by digital light and electron microscopy. The formation of ectoderm, endoderm and mesoderm and notochordal cells, and also the extraembryonic membranes, amnion and yolk sac, are imaged. The germ layers give rise to all the cells and tissues of the human body. Possible clues to the early differentiation of embryonic stem cells (ESC) in vitro were obtained, since these events are more or less mimicked in cultures of ESC derived from the inner cell mass of human blastocysts. The findings are discussed with reference to previous studies on the fine structure of ESC using the same technique. Copyright © 2011. Published by Elsevier Ltd.

  5. The treatment of tubal pregnancy by interventional technique through uterine artery

    International Nuclear Information System (INIS)

    Li Lizhao; Zhang Yonghui; Chen Zhu; Ge Xiaobing; Li Jiehua

    2004-01-01

    Objective: To study the feasibility and the clinical effect of the treatment of tubule pregnancy by using interventional technique through selective uterine artery. Methods: By using seldinger's method, 48 cases received interventional treatment, followed by perfusion of methotrexate (MTX) 100 mg through Tube 4-5 F. The concentration of serum β-HCG, the changes of pelvic cavity, and the open condition of amnion by ultrasonic examination. Results: The cure rate of 47 cases was 97.92%. No serious reaction. After treatment, the mean time that the serum β-HCG concentration returned to normal was 14-28 days and the mean time was 18 days. Conclusion: The treatment of tubule pregnancy by interventional technique proved no harmful effect to reproductive organs and quick recovery. It is worth spreading

  6. Morphological analysis of amnion stored in glycerol sterilized with different doses of ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Fernando Augusto N.; Santin, Stefany P.; Martino Junior, Antonio C.; Machado, Luci Diva B.; Mathor, Monica B., E-mail: fernandonevessoares@yahoo.com.br [Instituto de Pesquisas Energeticas e Nucleares (CTR/IPEN/CNEN-SP), Sao Paulo, RJ (Brazil). Centro de Tecnologia das Radiacoes

    2013-07-01

    The amniotic membrane (AM) is the innermost layer of the fetal membranes (placenta), widely used in transplantation being a tissue that combines anti-inflammatory, antimicrobial and antifibrotic, and limited immunogenicity. The tissue can be used a bandage biological for treatment of burns and skin wounds, chronic ulcers, reconstructions from different body areas, including ophthalmic repairs. In the last decades the amniotic membrane has been used widely also as a carrier substrate to transfer tissues cultured 'in vitro'. The use of fresh membrane has some limitations, the main ones are being necessary your quick use and the inability to obtain full security for certain infections. Other types of preservation require a processing thereof. The radiosterilization is an alternative for ensuring quality and safety of tissues used in transplants, and other clinical applications in order to minimize the risk of contamination of the receptor tissue. Therefore, the objective of this study was to test various doses of radiation using two sources of ionizing radiation: the cobalt-60 irradiator Gamma and Electron Beam Accelerator (E.B.). A tissue analysis was done by visual and tactile qualitative analysis, semi-quantitative (solid colorimetry) and light microscopy to observe morphological and physic-chemical changes after the irradiation of AM preserved in glycerol, comparing the results obtained with the sample not irradiated. It was noted that at higher doses, for both radiation sources, irradiated membranes suffered greater color change, becoming yellowish and thereby reducing their initial malleability. (author)

  7. Morphological analysis of amnion stored in glycerol sterilized with different doses of ionizing radiation

    International Nuclear Information System (INIS)

    Soares, Fernando Augusto N.; Santin, Stefany P.; Martino Junior, Antonio C.; Machado, Luci Diva B.; Mathor, Monica B.

    2013-01-01

    The amniotic membrane (AM) is the innermost layer of the fetal membranes (placenta), widely used in transplantation being a tissue that combines anti-inflammatory, antimicrobial and antifibrotic, and limited immunogenicity. The tissue can be used a bandage biological for treatment of burns and skin wounds, chronic ulcers, reconstructions from different body areas, including ophthalmic repairs. In the last decades the amniotic membrane has been used widely also as a carrier substrate to transfer tissues cultured 'in vitro'. The use of fresh membrane has some limitations, the main ones are being necessary your quick use and the inability to obtain full security for certain infections. Other types of preservation require a processing thereof. The radiosterilization is an alternative for ensuring quality and safety of tissues used in transplants, and other clinical applications in order to minimize the risk of contamination of the receptor tissue. Therefore, the objective of this study was to test various doses of radiation using two sources of ionizing radiation: the cobalt-60 irradiator Gamma and Electron Beam Accelerator (E.B.). A tissue analysis was done by visual and tactile qualitative analysis, semi-quantitative (solid colorimetry) and light microscopy to observe morphological and physic-chemical changes after the irradiation of AM preserved in glycerol, comparing the results obtained with the sample not irradiated. It was noted that at higher doses, for both radiation sources, irradiated membranes suffered greater color change, becoming yellowish and thereby reducing their initial malleability. (author)

  8. Transport and Biodistribution of Dendrimers Across Human Fetal Membranes: Implications for Intravaginal Administration of Dendrimers

    Science.gov (United States)

    Menjoge, Anupa R.; Navath, Raghavendra S.; Asad, Abbas; Kannan, Sujatha; Kim, Chong Jai; Romero, Roberto; Kannan, Rangaramanujam M.

    2010-01-01

    Dendrimers are emerging as promising topical antimicrobial agents, and as targeted nanoscale drug delivery vehicles. Topical intravaginal antimicrobial agents are prescribed to treat the ascending genital infections in pregnant women. The fetal membranes separate the extra-amniotic space and fetus. The purpose of the study is to determine if the dendrimers can be selectively used for local intravaginal application to pregnant women without crossing the membranes into the fetus. In the present study, the transport and permeability of PAMAM (poly(amidoamine)) dendrimers, across human fetal membrane (using a side-by-side diffusion chamber), and its biodistribution (using immunofluorescence) are evaluated ex-vivo. Transport across human fetal membranes (from the maternal side) was evaluated using Fluorescein (FITC), an established transplacental marker (positive control, size~ 400 Da) and fluorophore-tagged G4-PAMAM dendrimers (~ 16 kDa). The fluorophore-tagged G4-PAMAM dendrimers were synthesized and characterized using 1H NMR, MALDI TOF-MS and HPLC analysis. Transfer was measured across the intact fetal membrane (chorioamnion), and the separated chorion and amnion layers. Over a five hour period, the dendrimer transport across all the three membranes was less than transport of FITC was relatively fast with as much as 49% transport across the amnion. The permeability of FITC (7.9 × 10-7 cm2/s) through the chorioamnion was 7-fold higher than that of the dendrimer (5.8 × 10-8 cm2/s). The biodistribution showed that the dendrimers were largely present in interstitial spaces in the decidual stromal cells and the chorionic trophoblast cells (in 2.5 to 4 h) and surprisingly, to a smaller extent internalized in nuclei of trophoblast cells and nuclei and cytoplasm of stromal cells. Passive diffusion and paracellular transport appear to be the major route for dendrimer transport. The overall findings further suggest that entry of drugs conjugated to dendrimers would be

  9. Tissue banking and clinical research on radiation and ethylene oxide sterilization of tissue grafts

    International Nuclear Information System (INIS)

    Pe Khin

    1987-06-01

    The research works carried out in Rangoon, Burma under the Agency supported project RC4420/RB have dealt with an elucidation of the radiation interaction(s) with the species of biomolecules such as proteins, lipids, collagens, connective tissues present in the cleaned and freeze-dried non-viable tissue grafts. Radiation as a cool process furthermore effectively helps to destroy the microbial bioburden as the undesirable contaminants which may associate the tissue grafts. Radiation also concomitantly helps to suppress the tissue-specific immunogenicity. All these attributes of radiation induced effects have proved successful towards the development of a sterilization process. A series of non-viable tissue grafts, such as bone, nerve, fascia, dura, cartilage, chorion-amnion (as dressings in burn wounds) and tympanic membrane have been successfully attempted in Burma and many more possibilities seem to still remain unexplored. Radiation sterilization modality has proved as a blessing for the promotion of clinical surgical applications of tissue allografts in the corrective/reconstructive surgery on the disability cases due to diseases which accompany tissue losses. The investigator in Burma has reported on the case histories where freeze dried radiation sterilized tissue allografts have been successfully used in the osteogenic inductions (bone grafts); midear tympanoplasty; partial recovery of nerve sensation throught nerve allografts; rapid healing of high degree burn wounds through the use of amnion dressings. Besides, there have been a widespread surgical use of radiation sterilized dura and fascia as allografts. A national tissue banking facility has been established in Burma surrounding the processing and clinical utilization of tissue allografts which has involved over ten hospital centres throughout the country. Radiation induced effects on the biomolecules of clinical significance in the tissue grafts have been researched to help gain insight into a better

  10. Retinoid inhibition of in vitro invasion of human amnion basement membrane by human tumor cells

    International Nuclear Information System (INIS)

    Fazely, F.

    1988-01-01

    The effects measured were the inhibition of tumor cell migration through the basement membrane (BM) and tumor cell degradative enzyme activity on 3 H-proline labeled collagenous and non collagenous components of the BM. The human lung carcinoma A549 or the human Ewing's sarcoma TC-106 cell lines treated with retinoids for two days were incubated on the BM in the absence of retinoids. A dose-dependent inhibition of cell invasion was produced by retinoids. Among the retinoids tested the most powerful was retinol acetate which inhibited invasion by 50% of A549 cells at a concentration of 0.09 μg/ml, and TC-106 cells at 0.08 μg/ml. Retinol acetate inhibited A549 and TC-106 cell growth by approximately 50% at levels almost 100-fold higher than those needed for antiinvasive activity. Retinol acetate was about 20 times more potent than retinoic acid and 30 times more than retinol palmitate. Furthermore, A549 cells treated with retinol acetate, under conditions whereby an anti-invasive state was induced,showed an increase in the number of cellular retinoic acid binding proteins (CRABP), a decrease in the activity of type IV collagenase and ectosialyltransferase, and no change in the activity of transglutaminase

  11. [Perinatal sources of stem cells].

    Science.gov (United States)

    Piskorska-Jasiulewicz, Magdalena Maria; Witkowska-Zimny, Małgorzata

    2015-03-08

    Recently, stem cell biology has become an interesting topic. Several varieties of human stem cells have been isolated and identified in vivo and in vitro. Successful application of hematopoietic stem cells in hematology has led to the search for other sources of stem cells and expanding the scale of their application. Perinatal stem cells are a versatile cell population, and they are interesting for both scientific and practical objectives. Stem cells from perinatal tissue may be particularly useful in the clinic for autologous transplantation for fetuses and newborns, and after banking in later stages of life, as well as for in utero transplantation in the case of genetic disorders. In this review paper we focus on the extraction and therapeutic potential of stem cells derived from perinatal tissues such as the placenta, the amnion, amniotic fluid, umbilical cord blood and Wharton's jelly.

  12. Cell-mediated immunity to herpes simplex in humans: lymphocyte cytotoxicity measured by 51Cr release from infected cells

    International Nuclear Information System (INIS)

    Russell, A.S.; Percy, J.S.; Kovithavongs, T.

    1975-01-01

    We assessed cell-mediated immunity to herpes simplex virus type 1 antigen in patients suffering from recurrent cold sores and in a series of healthy controls. Paradoxically, all those subject to recurrent herpetic infections had, without exception, evidence of cell-mediated immunity to herpes antigens. This was demonstrated by lymphocyte transformation and specific 51 Cr release from infected human amnion cells after incubation with peripheral blood mononuclear cells. Where performed, skin tests with herpes antigen were also positive. In addition, serum from these patients specifically sensitized herpes virus-infected cells to killing by nonimmune, control mononuclear cells. These tests were negative in the control patients except in a few cases, and it is suggested that these latter may be the asymptomatic herpes virus carriers previously recognized or that they may have experienced a genital infection. (U.S.)

  13. Molecular cloning and expression of a novel keratinocyte protein (psoriasis-associated fatty acid-binding protein [PA-FABP]) that is highly up-regulated in psoriatic skin and that shares similarity to fatty acid-binding proteins

    DEFF Research Database (Denmark)

    Madsen, Peder; Rasmussen, H H; Leffers, H

    1992-01-01

    termed PA-FABP (psoriasis-associated fatty acid-binding protein). The deduced sequence predicted a protein with molecular weight of 15,164 daltons and a calculated pI of 6.96, values that are close to those recorded in the keratinocyte 2D gel protein database. The protein comigrated with PA-FABP...... as determined by 2D gel analysis of [35S]-methionine-labeled proteins expressed by transformed human amnion (AMA) cells transfected with clone 1592 using the vaccinia virus expression system and reacted with a rabbit polyclonal antibody raised against 2D gel purified PA-FABP. Structural analysis of the amino...... acid sequence revealed 48%, 52%, and 56% identity to known low-molecular-weight fatty acid-binding proteins belonging to the FABP family. Northern blot analysis showed that PA-FABP mRNA is indeed highly up-regulated in psoriatic keratinocytes. The transcript is present in human cell lines of epithelial...

  14. Perinatal sources of stem cells

    Directory of Open Access Journals (Sweden)

    Magdalena Maria Piskorska-Jasiulewicz

    2015-03-01

    Full Text Available Recently, stem cell biology has become an interesting topic. Several varieties of human stem cells have been isolated and identified in vivo and in vitro. Successful application of hematopoietic stem cells in hematology has led to the search for other sources of stem cells and expanding the scale of their application. Perinatal stem cells are a versatile cell population, and they are interesting for both scientific and practical objectives. Stem cells from perinatal tissue may be particularly useful in the clinic for autologous transplantation for fetuses and newborns, and after banking in later stages of life, as well as for in utero transplantation in the case of genetic disorders. In this review paper we focus on the extraction and therapeutic potential of stem cells derived from perinatal tissues such as the placenta, the amnion, amniotic fluid, umbilical cord blood and Wharton’s jelly.

  15. A "two-objective, one-area" procedure in absorption microphotometry and its application using an inverted microscope.

    Science.gov (United States)

    Chaubal, K A

    1988-08-01

    A 'two-objective, one-area' method and related equations are suggested to measure absorbance of microscopic stained objects. In such work, the measuring field invariably includes an image of the object and some clear area surrounding the image. The total intensity in the two areas is measured photometrically, using two different objectives, and substituted in the equation for absorbance. The equation is independent of the term representing intensity from the clear area and hence the error in the measurement of absorbance is reduced. The limitations of the 'two-objective, one-area' method are discussed and its pragmatic operation described with an experimental setup involving an inverted microscope. The method permits measurement of intensity in a part of a stained cell while the rest of the cell remains in the field of view. The method is applied to measure absorbance in Giemsa stained ascites cells and Feulgen stained liver and Human Amnion cells.

  16. The Consequences of Chorioamnionitis: Preterm Birth and Effects on Development

    Directory of Open Access Journals (Sweden)

    Robert Galinsky

    2013-01-01

    Full Text Available Preterm birth is a major cause of perinatal mortality and long-term morbidity. Chorioamnionitis is a common cause of preterm birth. Clinical chorioamnionitis, characterised by maternal fever, leukocytosis, tachycardia, uterine tenderness, and preterm rupture of membranes, is less common than subclinical/histologic chorioamnionitis, which is asymptomatic and defined by inflammation of the chorion, amnion, and placenta. Chorioamnionitis is often associated with a fetal inflammatory response. The fetal inflammatory response syndrome (FIRS is defined by increased systemic inflammatory cytokine concentrations, funisitis, and fetal vasculitis. Clinical and epidemiological studies have demonstrated that FIRS leads to poor cardiorespiratory, neurological, and renal outcomes. These observations are further supported by experimental studies that have improved our understanding of the mechanisms responsible for these outcomes. This paper outlines clinical and experimental studies that have improved our current understanding of the mechanisms responsible for chorioamnionitis-induced preterm birth and explores the cellular and physiological mechanisms underlying poor cardiorespiratory, neural, retinal, and renal outcomes observed in preterm infants exposed to chorioamnionitis.

  17. Molecular cloning and expression of a novel keratinocyte protein (psoriasis-associated fatty acid-binding protein [PA-FABP]) that is highly up-regulated in psoriatic skin and that shares similarity to fatty acid-binding proteins

    DEFF Research Database (Denmark)

    Madsen, Peder; Rasmussen, H H; Leffers, H

    1992-01-01

    termed PA-FABP (psoriasis-associated fatty acid-binding protein). The deduced sequence predicted a protein with molecular weight of 15,164 daltons and a calculated pI of 6.96, values that are close to those recorded in the keratinocyte 2D gel protein database. The protein comigrated with PA......-FABP as determined by 2D gel analysis of [35S]-methionine-labeled proteins expressed by transformed human amnion (AMA) cells transfected with clone 1592 using the vaccinia virus expression system and reacted with a rabbit polyclonal antibody raised against 2D gel purified PA-FABP. Structural analysis of the amino...... with epidermal growth factor (EGF), pituitary extract, and 10% fetal calf serum] revealed a strong up-regulation of PA-FABP, psoriasin, calgranulins A and B, and a few other proteins that are highly expressed in psoriatic skin. The levels of these proteins exceeded by far those observed in non-cultured normal...

  18. Treatment of burn injuries with keratinocyte cultures

    International Nuclear Information System (INIS)

    Syring, C.; Maenig, H.J.; Von Versen, R.; Bruck, J.

    1999-01-01

    The German Institute for Cell and Tissue Replacement (DIZG) provides burned patients with skin and amnion for a temporary wound closure. Severely burned patients (>60% BSA for adults, >40% BSA for children) were supplied with autologous and allogenic grafts from cultured keratinocytes. The keratinocyte culture is done under GMP-conditions using the method of Rheinwald and Green. The 3T3 fibroblasts were irradiated with 60 Gy and used as feeder cells to produce keratinocyte sheets within 3 weeks. In this time up to 6.000 cm are available. The sheets were harvested by detachment with dispase (1,2 U/ml), fixed to gauze and transported to the hospital. The DIZG has a 3 years experience in the treatment of burns with keratinocyte sheets. The sheets were transplanted to patients in different hospitals, the total transplanted area is about 30.000 cm. This paper describes the experiences with ten severely burned patients treated with keratinocyte sheet

  19. Is there a yet unreported unbalanced chromosomal abnormality without phenotypic consequences in proximal 4p?

    Science.gov (United States)

    Liehr, T; Bartels, I; Zoll, B; Ewers, E; Mrasek, K; Kosyakova, N; Merkas, M; Hamid, A B; von Eggeling, F; Posorski, N; Weise, A

    2011-01-01

    Unbalanced chromosomal abnormalities (UBCA) are reported for >50 euchromatic regions of almost all human autosomes. UBCA are comprised of a few megabases of DNA, and carriers are in many cases clinically healthy. Here we report on a partial trisomy of chromosome 4 of the centromere-near region of the short arm of chromosome 4 present as a small supernumerary marker chromosome (sSMC). The sSMC was present in >70% of amnion cells and in 60% of placenta. Further delineation of the size of the duplicated region was done by molecular cytogenetics and array comparative genomic hybridization. Even though the sSMC lead to a partial trisomy of ~9 megabase pairs, a healthy child was born, developing normally at 1 year of age. No comparable cases are available in the literature. Thus, we discuss here the possibility of having found a yet unrecognized chromosomal region subject to UBCA. Copyright © 2010 S. Karger AG, Basel.

  20. Possible Genetic Origin of Limb-Body Wall Complex.

    Science.gov (United States)

    Gajzer, David C; Hirzel, Alicia Cristina; Saigal, Gaurav; Rojas, Claudia Patricia; Rodriguez, Maria Matilde

    2015-01-01

    Limb body wall complex (LBWC) is characterized by multiple severe congenital malformations including an abdominal and/or thoracic wall defect covered by amnion, a short or absent umbilical cord with the placenta almost attached to the anterior fetal wall, intestinal malrotation, scoliosis, and lower extremity anomalies. There is no consensus about the etiology of LBWC and many cases with abnormal facial cleft do not meet the requirements for the true complex. We describe a series of four patients with LBWC and other malformations in an attempt to explain their etiology. There are several reports of fetuses with LBWC and absent gallbladder and one of our patients also had polysplenia. Absent gallbladder and polysplenia are associated with laterality genes including HOX, bFGF, transforming growth factor beta/activins/BMP4, WNT 1-8, and SHH. We postulate that this severe malformation may be due to abnormal genes involved in laterality and caudal development.

  1. Preparation of denatured protein bone sterilized with gamma radiation; Preparacion de hueso desproteinizado esterilizado con radiacion gamma

    Energy Technology Data Exchange (ETDEWEB)

    Luna Z, D [ININ, 52045 Ocoyoacac, Estado de Mexico (Mexico)

    2005-07-01

    The bone is one of the tissues more transplanted in the entire world by that the bone necessity for transplant every day becomes bigger. In the Bank of tissues Radio sterilized of the ININ the amnion and the pig skin are routinely processed. The tissue with which will be continued is with bone. Due to that in our country it doesn't have enough bone of human origin for the necessities required in the bone transplant, an option is the bone of bovine. Of this bone one can obtain denatured protein bone, with the same characteristics of the denatured protein human bone, the one which has been proven that it has good acceptance and incorporation in the human body when is transplanted. The method for the obtaining of the denatured protein bone of bovine, with the confirmation of the final product by means of X-ray diffraction is described. The radiosterilization of this bone with gamma rays and the determination of the lead content. (Author)

  2. Amniotic band syndrome (ABS): can something be done during pregnancy in African poor countries? Three cases and review of the literature.

    Science.gov (United States)

    Mian, D B; Nguessan, K L P; Aissi, G; Boni, S

    2014-01-01

    Amniotic band syndrome (ABS) is a fetal congenital malformation, affecting mainly the limbs, but also the craniofacial area and internal organs. Two mains pathogenic mechanisms are proposed in its genesis. Firstly the early amnion rupture (exogenous theory) leading to fibrous bands, which wrap up the fetal body; secondly, the endogenous theory privileges vascular origin, mesoblastic strings not being a causal agent. The authors believe that the second theory explain the occurrence of ABS. The outcome of the disease during pregnancy depends on the gravity of the malformations. Interruption of the pregnancy is usually proposed when diagnosis of severe craniofacial and visceral abnormalities is confirmed. Whereas minor limb defects can be repaired with postnatal surgery. In case of an isolated amniotic band with a constricted limb, in utero lysis of the band can be considered to avoid a natural amputation. In an African country, such treatment is not possible as far as the antenatal diagnosis.

  3. Mechanisms leading to increased risk of preterm birth in growth-restricted guinea pig pregnancies.

    Science.gov (United States)

    Palliser, Hannah K; Kelleher, Meredith A; Welsh, Toni N; Zakar, Tamas; Hirst, Jonathan J

    2014-02-01

    Intrauterine growth restriction (IUGR) is a risk factor for preterm labor; however, the mechanisms of the relationship remain unknown. Prostaglandin (PG), key stimulants of labor, availability is regulated by the synthetic enzymes, prostaglandin endoperoxidases 1 and 2 (PTGS1 and 2), and the metabolizing enzyme, 15-hydroxyprostaglandin dehydrogenase (HPGD). We hypothesized that IUGR increases susceptibility to preterm labor due to the changing balance of synthetic and metabolizing enzymes and hence greater PG availability. We have tested this hypothesis using a surgically induced IUGR model in guinea pigs, which results in significantly shorter gestation. Myometrium, amnion, chorion, and placentas were collected from sham operated or IUGR pregnancies, and PTGS1 and HPGD protein expression were quantified throughout late gestation (>62 days) and labor. The PTGS1 expression was significantly upregulated in the myometrium of IUGR animals, and chorionic HPGD expression was markedly decreased (P production over metabolism in IUGR pregnancies leads to a greater susceptibility to preterm birth.

  4. Preterm labor and preterm premature rupture of membranes have a different pattern in the involved compartments of acute histologoic chorioamnionitis and/or funisitis: Patho-physiologic implication related to different clinical manifestations.

    Science.gov (United States)

    Park, Chan-Wook; Park, Joong Shin; Moon, Kyung Chul; Jun, Jong Kwan; Yoon, Bo Hyun

    2016-06-01

    It is unknown whether histo-topographic findings about the involved compartments (i.e., choriodecidua, amnion, chorionic-plate) of acute-histologic chorioamnionitis (acute-HCA) and/or funisitis according to the presence or absence of intra-amniotic inflammation (IAI) and/or fetal inflammatory response syndrome (FIRS) are different between preterm labor and intact membranes (PTL) and preterm premature rupture of membranes (preterm-PROM). The involved compartments of acute-HCA and/or funisitis were examined in 161 singleton preterm-births ( 0.1). However, IAI(+)/FIRS(+) group had a significantly higher rate of inflammation in each compartment than IAI(+)/FIRS(-) group in both PTL and preterm-PROM (each-for P < 0.05). We first demonstrated that PTL and preterm-PROM had a different pattern in the involved compartments of acute-HCA and/or funisitis in the IAI(-)/FIRS(--) group and in the change of involved compartments from IAI(-)/FIRS(-) to IAI(+)/FIRS(-). © 2016 Japanese Society of Pathology and John Wiley & Sons Australia, Ltd.

  5. Preparation of denatured protein bone sterilized with gamma radiation; Preparacion de hueso desproteinizado esterilizado con radiacion gamma

    Energy Technology Data Exchange (ETDEWEB)

    Luna Z, D. [ININ, 52045 Ocoyoacac, Estado de Mexico (Mexico)]. e-mail: dlz@nuclear.inin.mx

    2005-07-01

    The bone is one of the tissues more transplanted in the entire world by that the bone necessity for transplant every day becomes bigger. In the Bank of tissues Radio sterilized of the ININ the amnion and the pig skin are routinely processed. The tissue with which will be continued is with bone. Due to that in our country it doesn't have enough bone of human origin for the necessities required in the bone transplant, an option is the bone of bovine. Of this bone one can obtain denatured protein bone, with the same characteristics of the denatured protein human bone, the one which has been proven that it has good acceptance and incorporation in the human body when is transplanted. The method for the obtaining of the denatured protein bone of bovine, with the confirmation of the final product by means of X-ray diffraction is described. The radiosterilization of this bone with gamma rays and the determination of the lead content. (Author)

  6. Preparation of denatured protein bone sterilized with gamma radiation

    International Nuclear Information System (INIS)

    Luna Z, D.

    2005-01-01

    The bone is one of the tissues more transplanted in the entire world by that the bone necessity for transplant every day becomes bigger. In the Bank of tissues Radio sterilized of the ININ the amnion and the pig skin are routinely processed. The tissue with which will be continued is with bone. Due to that in our country it doesn't have enough bone of human origin for the necessities required in the bone transplant, an option is the bone of bovine. Of this bone one can obtain denatured protein bone, with the same characteristics of the denatured protein human bone, the one which has been proven that it has good acceptance and incorporation in the human body when is transplanted. The method for the obtaining of the denatured protein bone of bovine, with the confirmation of the final product by means of X-ray diffraction is described. The radiosterilization of this bone with gamma rays and the determination of the lead content. (Author)

  7. Elaboration of amniotic membrane dressing dried by air and irradiated - Peruvian experience

    International Nuclear Information System (INIS)

    Gamero, E.C.; Perez Caballero, N.

    1999-01-01

    The purpose of this work is to prepare dressings from the amniotic membrane to be used in cases of skin damage principally due to superficial and intermediate second-degree burns. The amnion is a transparent membrane that lines the chorion. It is resistant and rich in collagen. Due to these characteristics it can be well used as biological dressing as it diminishes the loss of fluids, electrolytes and proteins, it also protects the growing epithelium and adheres well to the surface of the wound, improves mobility of the patient, diminishing pain and stimulating neovascularization. The ISN-IPEN Tissue Bank promoted by IAEA has processed amniotic membrane since July 1997. Initially dressings were prepared using antibiotics, after IAEA training at the MINT of Malaysia, it is processed dried by air, lyophylized and in both presentations, sterilized by gamma-rays. Amniotic membranes are procured from Lima Maternity. Tissues must comply with VDRL, HIV, Hepatitis B and C exclusion tests. The process is held in a laminar flow hood and amnion already separated from the chorion is washed with sterile distilled water, a solution of 0.05% sodium hypochlorite, and normal saline. Then it is cut into appropriate sizes and double packed in PE films. The dressings are then carried to the Peruvian Institute of Nuclear Energy for irradiation, depending on the number of samples either irradiated with gamma-rays at the Gammacell 220 or at the Irradiation Facility located in Santa Anita. The delivered dose is 25 kGy. The product is only released if it complies with the end product quality controls. Meanwhile, microbiological tests are carried out during all the processing stages, in order to monitor the microbial load during production. In conclusion we can state that dressings prepared as above mentioned have the following advantages: not complicated preparation; reliable and safe for clinical use; diminish infection rates and days spent in the hospital; easy to storage; and can be

  8. RESEARCH ON REDUCING PREMATURITY RUPTURE OF MEMBRANE

    Directory of Open Access Journals (Sweden)

    Maria URSACHI (BOLOTA

    2016-12-01

    Full Text Available The membranes surrounding the amniotic cavity are composed from amnion and chorion, tightly adherent layers which are composed of several cell types, including epithelial cells, trophoblasts cells and mesenchyme cells, embedded in a collagenous matrix. They retain amniotic fluid, secret substances into the amniotic fluid, as well as to the uterus and protect the fetus against upward infections from urogenital tract. Normally, the membranes it breaks during labor. Premature rupture of the amniotic sac (PRAS is defined as rupture of membranes before the onset of labor. Premature rupture of the fetal membrane, which occurs before 37 weeks of gestation, usually, refers to preterm premature rupture of membranes. Despite advances in the care period, premature rupture of membranes and premature rupture of membranes preterm continue to be regarded as serious obstetric complications. On the term 8% - 10% of pregnant women have premature rupture of membranes; these women are at increased risk of intrauterine infections, where the interval between membrane rupture and expulsion is rolled-over. Premature rupture of membranes preterm occurs in approximately 1% of all pregnancies and is associated with 30% -40% of preterm births. Thus, it is important to identify the cause of pre-term birth (after less than 37 completed weeks of "gestation" and its complications, including respiratory distress syndrome, neonatal infection and intraventricular hemorrhage. Objectives: the development of the protocol of the clinical trial on patients with impending preterm birth, study clinical and statistical on the socio-demographic characteristics of patients with imminent preterm birth; clinical condition of patients and selection of cases that could benefit from the application of interventional therapy; preclinical investigation (biological and imaging of patients with imminent preterm birth; the modality therapy; clinical investigation of the effectiveness of short

  9. Caracterização das membranas fetais em búfalas no terço inicial da gestação Characterization of the fetal membranes in buffalo cows in the first three months of gestation

    Directory of Open Access Journals (Sweden)

    Adriana C. Morini

    2008-09-01

    Full Text Available Este estudo visou à caracterização das membranas fetais em búfalas (Bubalus bubalis, Linnaeus 1758 no terço inicial da gestação. As membranas fetais foram analisadas macroscópica e microscopicamente (luz e microscopia eletrônica de transmissão. O córion possui uma camada simples de células circulares, com núcleos de forma esférica, denominadas trofobláticas; há outro tipo celular, as células trofoblásticas gigantes, com dois ou mais núcleos. Ambas possuem uma grande quantidade de vesículas no citoplasma e retículo endoplasmático à microscopia de transmissão. O alantóide possui vasos preenchidos com eritrócitos, e contêm células alongadas, que formam um epitélio estratificado simples. O âmnion é uma membrana transparente, ou esbranquiçada; constituído por epitélio estratificado simples. A diferença principal entre o alantóide e o âmnion é que o último é avascular. O saco vitelínico é uma membrana opaca que desaparece durante a gestação; é a única membrana que não está em contato com as outras e apresenta três tipos diferentes de células que dão forma a três camadas distintas (endoderma, mesotélio, mesênquima.The aim of the study was to characterize the fetal membranes in buffalo cows (Bubalus bubalis, Linnaeus 1758 in the first 3 months of the gestation. The fetal membranes were analyzed gross and microscopically (light and ultrastructural analysis. The corion is a simple layer of circle cells, with spherical nucleus, named trofoblastics; there is another cellular type, named trophoblastic giant cells, with two or more nucleus. Both of them have many cytoplasmic and reticule endoplasmic vesicles on transmission electron microscopy. The allantoic has many vessels filled with eritrocytes, elongated cells which form a stratified simple epithelium. The amnion is transparent or sometimes whitish, consisting of a stratified simple epithelium. The main difference between the allantoic and amnion is

  10. Injectible candidate sealants for fetal membrane repair: Bonding and toxicity in vitro

    Science.gov (United States)

    Bilic, Grozdana; Brubaker, Carrie; Messersmith, Phillip B.; Mallik, Ajit S.; Quinn, Thomas M.; Haller, Claudia; Done, Elisa; Gucciardo, Leonardo; Zeisberger, Steffen M.; Zimmermann, Roland; Deprest, Jan; Zisch, Andreas H.

    2010-01-01

    Objective This study was undertaken to test injectible surgical sealants that are biocompatible with fetal membranes, eventually for closure of iatrogenic membrane defects. Study Design Dermabond, Histoacryl, Tissucol fibrin glue, and three types of in situ forming poly(ethylene glycol)-based polymer hydrogels were tested for acute toxicity upon direct contact with fetal membranes for 24h. For determination of elution toxicity, extracts of sealants were incubated on amnion cell cultures for 72h. Bonding and toxicity was assessed through morphological and/or biochemical analysis. Results Extracts of all adhesives were non-toxic for cultured cells. However, only Tissucol and one type of poly(ethylene glycol)-based hydrogel, mussel-mimetic tissue adhesive, showed efficient, non-disruptive, non-toxic bonding to fetal membranes. Mussel-mimetic tissue adhesive applied over membrane defects created with a 3.5 mm trocar accomplished leak-proof closure that withstood membrane stretch in an in vitro model. Conclusion A synthetic hydrogel-type tissue adhesive emerged as potential sealing modality for iatrogenic membrane defects that merits further evaluation in vivo. PMID:20096254

  11. Postnatal and antenatal laboratory diagnosis of glutaric aciduria II in a South African family

    International Nuclear Information System (INIS)

    Henderson, H.E.; Balla, R.; De Jong, G.; Piek, C.J.; Mienie, L.J.; Erasmus, E.

    1987-01-01

    Glutaric aciduria type II (Ga II) was proved in a neonate who presented shortly after birth with respiratory distress, metabolic acidosis, non-ketotic hypoglycaemia and a sweaty-feet-like odour. The diagnosis was based on elevated levels of glutaric and other acids in the urine and on studies on cultured skin fibroblasts where defective metabolism of fatty acids of varying chain length was demonstrated. Antenatal diagnosis was performed on a subsequent pregnancy in this family where an abnormal amniotic fluid organic acid profile together with defective fatty acid oxidation in cultured amnion cells was indicative of GA II in the fetus. A measure of efficency of butyrate oxidation was obtained by analysis of the amount of 14 C label incorporated into trichloro-acetic acid precipitable macromolecules. The incorporation of ( 3 H) leucine was taken as a general indicator of metabolism in the cell population. This is the first report of this genetic disorder in a South African family and it should be considered in suspected organic acidaemia in the neonatal period

  12. Challenges in validating the sterilisation dose for processed human amniotic membranes

    International Nuclear Information System (INIS)

    Yusof, Norimah; Hassan, Asnah; Firdaus Abd Rahman, M.N.; Hamid, Suzina A.

    2007-01-01

    Most of the tissue banks in the Asia Pacific region have been using ionising radiation at 25 kGy to sterilise human tissues for save clinical usage. Under tissue banking quality system, any dose employed for sterilisation has to be validated and the validation exercise has to be a part of quality document. Tissue grafts, unlike medical items, are not produced in large number per each processing batch and tissues relatively have a different microbial population. A Code of Practice established by the International Atomic Energy Agency (IAEA) in 2004 offers several validation methods using smaller number of samples compared to ISO 11137 (1995), which is meant for medical products. The methods emphasise on bioburden determination, followed by sterility test on samples after they were exposed to verification dose for attaining of sterility assurance level (SAL) of 10 -1 . This paper describes our experience in using the IAEA Code of Practice in conducting the validation exercise for substantiating 25 kGy as sterilisation dose for both air-dried amnion and those preserved in 99% glycerol

  13. Challenges in validating the sterilisation dose for processed human amniotic membranes

    Energy Technology Data Exchange (ETDEWEB)

    Yusof, Norimah [Malaysian Nuclear Agency, Bangi, 43000 Kajang, Selangor (Malaysia)], E-mail: norimah@mint.gov.my; Hassan, Asnah [Malaysian Nuclear Agency, Bangi, 43000 Kajang, Selangor (Malaysia); Firdaus Abd Rahman, M.N.; Hamid, Suzina A. [National Tissue Bank, Hospital Universiti Sains Malaysia, Kubang Kerian, 16130 Kelantan (Malaysia)

    2007-11-15

    Most of the tissue banks in the Asia Pacific region have been using ionising radiation at 25 kGy to sterilise human tissues for save clinical usage. Under tissue banking quality system, any dose employed for sterilisation has to be validated and the validation exercise has to be a part of quality document. Tissue grafts, unlike medical items, are not produced in large number per each processing batch and tissues relatively have a different microbial population. A Code of Practice established by the International Atomic Energy Agency (IAEA) in 2004 offers several validation methods using smaller number of samples compared to ISO 11137 (1995), which is meant for medical products. The methods emphasise on bioburden determination, followed by sterility test on samples after they were exposed to verification dose for attaining of sterility assurance level (SAL) of 10{sup -1}. This paper describes our experience in using the IAEA Code of Practice in conducting the validation exercise for substantiating 25 kGy as sterilisation dose for both air-dried amnion and those preserved in 99% glycerol.

  14. Discovery of HeLa Cell Contamination in HES Cells: Call for Cell Line Authentication in Reproductive Biology Research.

    Science.gov (United States)

    Kniss, Douglas A; Summerfield, Taryn L

    2014-08-01

    Continuous cell lines are used frequently in reproductive biology research to study problems in early pregnancy events and parturition. It has been recognized for 50 years that many mammalian cell lines contain inter- or intraspecies contaminations with other cells. However, most investigators do not routinely test their culture systems for cross-contamination. The most frequent contributor to cross-contamination of cell lines is the HeLa cell isolated from an aggressive cervical adenocarcinoma. We report on the discovery of HeLa cell contamination of the human endometrial epithelial cell line HES isolated in our laboratory. Short tandem repeat analysis of 9 unique genetic loci demonstrated molecular identity between HES and HeLa cells. In addition, we verified that WISH cells, isolated originally from human amnion epithelium, were also contaminated with HeLa cells. Inasmuch as our laboratory did not culture HeLa cells at the time of HES cell derivations, the source of contamination was the WISH cell line. These data highlight the need for continued diligence in authenticating cell lines used in reproductive biology research. © The Author(s) 2014.

  15. Inhibition of Various Cancer Cells Proliferation of Bornyl Acetate and Essential Oil from Inula graveolens (Linnaeus Desf

    Directory of Open Access Journals (Sweden)

    Tunay Karan

    2018-05-01

    Full Text Available Inula species are medicinal and aromatic plants used for folk medicine extensively. In this work, hydrodistilled essential oil of Inula graveolens (Linnaeus Desf. analyzed by GC-MS which revealed that bornyl acetate was the major product (69.15%. Camphene was the second major compound (11.11%. Antiproliferative activity of the essential oil and bornyl acetate was investigated on HeLa (human cervix carcinoma, HT29 (human colon carcinoma , A549 (human lung carcinoma, MCF7 ( human breast adenocarcinoma cancer cells and FL (human amnion cells normal cells. The cytotoxicity was executed by a Lactate Dehydrogenase (LDH Cytotoxicity Detection Kit. Essential oil and bornyl acetate displayed the outstanding activities on HeLa (IC 50, 64.1, 72.0 µg/mL, HT29 (IC 50, 24.6, 60.5 µg/mL, A549 (IC 50, 28.3, 44.1 µg/mL, MCF-7 (IC 50, 66.5, 85.6 µg/mL, and FL (IC 50, 42.1, 50.6 µg/mL cell lines respectively.

  16. Evidence for increased SOX3 dosage as a risk factor for X-linked hypopituitarism and neural tube defects.

    Science.gov (United States)

    Bauters, Marijke; Frints, Suzanna G; Van Esch, Hilde; Spruijt, Liesbeth; Baldewijns, Marcella M; de Die-Smulders, Christine E M; Fryns, Jean-Pierre; Marynen, Peter; Froyen, Guy

    2014-08-01

    Genomic duplications of varying lengths at Xq26-q27 involving SOX3 have been described in families with X-linked hypopituitarism. Using array-CGH we detected a 1.1 Mb microduplication at Xq27 in a large family with three males suffering from X-linked hypopituitarism. The duplication was mapped from 138.7 to 139.8 Mb, harboring only two annotated genes, SOX3 and ATP11C, and was shown to be a direct tandem copy number gain. Unexpectedly, the microduplication did not fully segregate with the disease in this family suggesting that SOX3 duplications have variable penetrance for X-linked hypopituitarism. In the same family, a female fetus presenting with a neural tube defect was also shown to carry the SOX3 copy number gain. Since we also demonstrated increased SOX3 mRNA levels in amnion cells derived from an unrelated t(X;22)(q27;q11) female fetus with spina bifida, we propose that increased levels of SOX3 could be a risk factor for neural tube defects. © 2014 Wiley Periodicals, Inc.

  17. Onset of human preterm and term birth is related to unique inflammatory transcriptome profiles at the maternal fetal interface

    Directory of Open Access Journals (Sweden)

    Radek Bukowski

    2017-09-01

    Full Text Available Background Preterm birth is a main determinant of neonatal mortality and morbidity and a major contributor to the overall mortality and burden of disease. However, research of the preterm birth is hindered by the imprecise definition of the clinical phenotype and complexity of the molecular phenotype due to multiple pregnancy tissue types and molecular processes that may contribute to the preterm birth. Here we comprehensively evaluate the mRNA transcriptome that characterizes preterm and term labor in tissues comprising the pregnancy using precisely phenotyped samples. The four complementary phenotypes together provide comprehensive insight into preterm and term parturition. Methods Samples of maternal blood, chorion, amnion, placenta, decidua, fetal blood, and myometrium from the uterine fundus and lower segment (n = 183 were obtained during cesarean delivery from women with four complementary phenotypes: delivering preterm with (PL and without labor (PNL, term with (TL and without labor (TNL. Enrolled were 35 pregnant women with four precisely and prospectively defined phenotypes: PL (n = 8, PNL (n = 10, TL (n = 7 and TNL (n = 10. Gene expression data were analyzed using shrunken centroid analysis to identify a minimal set of genes that uniquely characterizes each of the four phenotypes. Expression profiles of 73 genes and non-coding RNA sequences uniquely identified each of the four phenotypes. The shrunken centroid analysis and 10 times 10-fold cross-validation was also used to minimize false positive finings and overfitting. Identified were the pathways and molecular processes associated with and the cis-regulatory elements in gene’s 5′ promoter or 3′-UTR regions of the set of genes which expression uniquely characterized the four phenotypes. Results The largest differences in gene expression among the four groups occurred at maternal fetal interface in decidua, chorion and amnion. The gene expression profiles showed

  18. A Human Amnion-Derived Extracellular Matrix-Coated Cell-Free Scaffold for Cartilage Repair: In Vitro and In Vivo Studies.

    Science.gov (United States)

    Nogami, Makiko; Kimura, Tomoatsu; Seki, Shoji; Matsui, Yoshito; Yoshida, Toshiko; Koike-Soko, Chika; Okabe, Motonori; Motomura, Hiraku; Gejo, Ryuichi; Nikaido, Toshio

    2016-04-01

    Extracellular matrix (ECM) derived from human amniotic mesenchymal cells (HAMs) has various biological activities. In this study, we developed a novel HAM-derived ECM-coated polylactic-co-glycolic acid (ECM-PLGA) scaffold, examined its property on mesenchymal cells, and investigated its potential as a cell-free scaffold for cartilage repair. ECM-PLGA scaffolds were developed by inoculating HAM on a PLGA. After decellularization by irradiation, accumulated ECM was examined. Exogenous cell growth and differentiation of rat mesenchymal stem cells (MSCs) on the ECM-PLGA were analyzed in vitro by cell attachment/proliferation assay and reverse transcription-polymerase chain reaction. The cell-free ECM-PLGA scaffolds were implanted into osteochondral defects in the trochlear groove of rat knees. After 4, 12, or 24 weeks, the animals were sacrificed and the harvested tissues were examined histologically. The ECM-PLGA contained ECM that mimicked natural amniotic stroma that contains type I collagen, fibronectin, hyaluronic acid, and chondroitin sulfates. The ECM-PLGA showed excellent properties of cell attachment and proliferation. MSCs inoculated on the ECM-PLGA scaffold showed accelerated type II collagen mRNA expression after 3 weeks in culture. The ECM-PLGA implanted into an osteochondral defect in rat knees induced gradual tissue regeneration and resulted in hyaline cartilage repair, which was better than that in the empty control group. These in vitro and in vivo experiments show that the cell-free scaffold composed of HAM-derived ECM and PLGA provides a favorable growth environment for MSCs and facilitates the cartilage repair process. The ECM-PLGA may become a "ready-made" biomaterial for cartilage repair therapy.

  19. Prenatal ultrasound diagnosis of omphalocele

    International Nuclear Information System (INIS)

    Rio Romero, Luskenia del; Blanco Figueredo, Nadia; Rodriguez Dominguez, Zulay

    2014-01-01

    Omphalocele is an abdominal wall defect at the midline characterized by herniation of abdominal contents and covered by peritoneum and amnion. The aim of this paper is to present a case of omphalocele with gestational age of 23 weeks and prenatal diagnosis by ultrasonography. Using ultrasound diagnosis in a patient inquest made 40 years of age in the second trimester (gestational age 23 weeks) showed a level of the anterior fetal echogenic image that sticks through the abdominal wall and then locate the cord umbilical. Stomach is seen displaced and loss of normal anatomy of the abdominal circumference. Genetic counseling was conducted at the Municipal Center for Genetics of Manzanillo. Pathologically the fetus presented short and wide neck, low-set ears, defect omphalomesenteric of ductal closure, hernia sac occupied by the caudate lobe of the liver and gallbladder bed, wide base heart dissection showing cava-cava absence of interventricular septum was observed pulmonary valve stenosis most dilation of supravalvular pulmonary artery, large defect and aorta intraventricular septum ride, which speaks in favor of a heart rate troncoconal fallop trilogy over the omphalocele. Prenatal diagnosis by ultrasonography is an efficient and reliable method for prenatal diagnosis of omphalocele

  20. Effect of substratum, serum and linoleic acid on the lipid synthesis of isolated alveolar type II cells

    International Nuclear Information System (INIS)

    Cott, G.R.; Edeen, K.E.; Hale, S.G.; Mason, R.J.

    1986-01-01

    The authors examined the effect of cellular substratum (plastic or amnionic basement membrane (ABM)) and serum additive (fetal bovine (FBS), pork, horse, rat or human) on phospholipid synthesis in alveolar type II cells. The cells were isolated from adult rats, cultured for 48 hours under the various substratum and serum conditions, and then incubated for an additional 2 hours with [1- 14 C] acetate. ABM consistently caused a significant increase in the percent of radiolabel incorporated into phosphatidylcholine (PC) and/or phosphatidylglycerol (PG). Serum also had a significant effect with the highest values of PC and saturated PC being obtained with rat serum and the highest PG values with horse serum. The fatty acid composition of the sera used varied according to species with the largest variations in percent linoleic acid. Supplementing media with linoleic acid resulted in a marked increase in saturated PC values and a fall in PG values. Therefore, they conclude that: 1) ABM improves differentiated function, 2) FBS supplementation may not be optimal, and 3) the different effects of linoleic acid supplementation on PC, saturated PC, and PG values suggests an independent regulation of synthesis for these lipid species in vitro

  1. A tensile machine with a novel optical load cell for soft biological tissues application.

    Science.gov (United States)

    Faturechi, Rahim; Hashemi, Ata; Abolfathi, Nabiollah

    2014-11-01

    The uniaxial tensile testing machine is the most common device used to measure the mechanical properties of industrial and biological materials. The need for a low-cost uniaxial tension testing device for small research centers has always been the subject of research. To address this need, a novel uniaxial tensile testing machine was designed and fabricated to measure the mechanical properties of soft biological tissues. The device is equipped with a new low-cost load cell which works based on the linear displacement/force relationship of beams. The deflection of the beam load cell is measured optically by a digital microscope with an accuracy of 1 µm. The stiffness of the designed load cell was experimentally and theoretically determined at 100 N mm(-1). The stiffness of the load cell can be easily adjusted according to the tissue's strength. The force-time behaviour of soft tissue specimens was obtained by an in-house image processing program. To demonstrate the efficiency of the fabricated device, the mechanical properties of amnion tissue was measured and compared with available data. The obtained results indicate a strong agreement with that of previous studies.

  2. The expression and function of fatty acid transport protein-2 and -4 in the murine placenta.

    Directory of Open Access Journals (Sweden)

    Takuya Mishima

    Full Text Available The uptake and trans-placental trafficking of fatty acids from the maternal blood into the fetal circulation are essential for embryonic development, and involve several families of proteins. Fatty acid transport proteins (FATPs uniquely transport fatty acids into cells. We surmised that placental FATPs are germane for fetal growth, and are regulated during hypoxic stress, which is associated with reduced fat supply to the fetus.Using cultured primary term human trophoblasts we found that FATP2, FATP4 and FATP6 were highly expressed in trophoblasts. Hypoxia enhanced the expression of trophoblastic FATP2 and reduced the expression of FATP4, with no change in FATP6. We also found that Fatp2 and Fatp4 are expressed in the mouse amnion and placenta, respectively. Mice deficient in Fatp2 or Fatp4 did not deviate from normal Mendelian distribution, with both embryos and placentas exhibiting normal weight and morphology, triglyceride content, and expression of genes related to fatty acid mobilization.We conclude that even though hypoxia regulates the expression of FATP2 and FATP4 in human trophoblasts, mouse Fatp2 and Fatp4 are not essential for intrauterine fetal growth.

  3. ASPHYXIA, INTRACRANIAL HEMORRHAGES AND BRAIN EDEMA OF RISK CHILDREN IN THE ADVISORY INSTITUTE IN BITOLA FROM 1989-1994

    Directory of Open Access Journals (Sweden)

    M. ILIEVSKA,

    1997-09-01

    Full Text Available 3986 files have been examined in the Advisory Institute for a five year period in relation to the present risk factors in the pre, peri and postnatal period, the occurrence of asphyxia, I.H. (intracranial hemorrhages and brain edema and their outcome for the children. There were 958 or 32% risk children, out of them 206 or 22% were with asphyxia, 25 or 3% were with brain edema and 14 or 1,5% were with intracranial hemorrhages.The analysis for the risk factors shows that 119 of them were abortive , and from them 15% were born with asphyxia; 124 were SFD and 21% of them with asphyxia; 272 children weighed over 4500 gr., 7% of them with asphyxia and 0.4% with I.H., there were 68 twins, 12% of them with asphyxia. Out of the children with no risk registered, 6 were born with I.H., or 0,2%.Mothers under the age of 18 gave birth to 13% children with asphyxia; treated for sterility and anemia during pregnancy 15%; with increased blood pressure 14%; and 5% with maintained pregnancy.The highest delivery risk is present with children born with vacuum extraction (30% or every third child is with asphyxia and 3% with I.H. and with children delivered by caesarean section (14% with asphyxia.As for the position of the fetus-Citus pedalicus gave 55% children with asphyxia, and Situs pelvicus 12%.The worst damage is suffered by infants with premature amnion disruption (62% are with asphyxia; with the umbilical cord round the neck-56% with asphyxia and 6% with I.H.; and with muddled amniotic fluid and placenta pelvia-50%.The order of risk factors related to asphyxia, I.H. and brain edema is as follows: the first is premature amnion disruption, then follows the umbilical cord round the neck, the muddled amniotic fluid, and placenta previa and Citus pedalicus-which are obstetric problems. The next are the vacuum extraction and S.C. As for the gestatory period the order is as follows: first the abortive, then the twins and hypertrofic infants. The outcome of the

  4. Evolução morfométrica dos anexos embrionários e fetais bovinos obtidos por monta natural, com 10 a 70 dias da gestação Biometrics evolution of the embryonic and fetal annexes in cows obtained by natural mating, at 10 to 70 days of gestation

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    Antônio C. de Assis Neto

    2009-10-01

    Full Text Available O período inicial da gestação de bovinos é caracterizado por grandes perdas embrionárias. Considerando a importância deste fator no âmbito da reprodução animal foram estudados os anexos embrionários e fetais bovinos fecundados por monta natural de 15-70 dias de gestação, com o objetivo de estabelecer parâmetros morfométricos da placenta na fase inicial da gestação. Com uso de um paquímetro foram realizadas mensurações do comprimento (crânio caudal, largura (latero lateral e altura (dorso ventral das membranas corioalantóide e amniótica. O início da formação dos cotilédones foi observado e quantificado, assim como, o peso placentário. O peso médio do saco gestacional aumentou com o evoluir da idade gestacional, entretanto, o crescimento foi acelerado a partir de 20-30 dias de gestação. O comprimento crânio caudal e dorso ventral da membrana corioalantóide e do âmnio apresentaram crescimento lento e gradual com o evoluir dos períodos gestacionais analisados. Com 30-40 dias de gestação, os primeiro cotilédones já eram visualizados e contatos com facilidade na superfície coriônica. Os períodos de crescimento coincidiram com os maiores índices de perdas gestacionais em bovinos. Os parâmetros aqui analisados poderão servir para futuras investigações dos anexos embrionários de organismos manipulados em laboratório.The main goal of this morphometrical study was to characterize the development of the extra-embryonic membranes of 15 to 70-day-old bovine embryos obtained by natural mating. With a millimeter paquimeter the cranio-caudal, latero-lateral and dorso-ventral measurements of chorion and amnion were determined. The development of the cotyledons and weight of the gestational sac were observed and quantified. The weight of the gestational sac increased during gestation; however, the growth was faster then 20 to 30 days. The cranial-caudal and dorsal-ventral length of the choriallantois and the amnion

  5. Combination of hydroxyapatite, platelet rich fibrin and amnion membrane as a novel therapeutic option in regenerative periapical endodontic surgery: Case series

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    Uday Kiran Uppada

    2017-01-01

    Conclusion: The results of this case seriessubstantiatesthe credibility of using a combination ofamnion membrane with a bone graft and PRF to enhance radiographic healing outcome with decreased post-operative discomfort and present a viable regenerative treatment modality in periapical surgery.

  6. DNA methylation dynamics in human induced pluripotent stem cells over time.

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    Koichiro Nishino

    2011-05-01

    Full Text Available Epigenetic reprogramming is a critical event in the generation of induced pluripotent stem cells (iPSCs. Here, we determined the DNA methylation profiles of 22 human iPSC lines derived from five different cell types (human endometrium, placental artery endothelium, amnion, fetal lung fibroblast, and menstrual blood cell and five human embryonic stem cell (ESC lines, and we followed the aberrant methylation sites in iPSCs for up to 42 weeks. The iPSCs exhibited distinct epigenetic differences from ESCs, which were caused by aberrant methylation at early passages. Multiple appearances and then disappearances of random aberrant methylation were detected throughout iPSC reprogramming. Continuous passaging of the iPSCs diminished the differences between iPSCs and ESCs, implying that iPSCs lose the characteristics inherited from the parent cells and adapt to very closely resemble ESCs over time. Human iPSCs were gradually reprogrammed through the "convergence" of aberrant hyper-methylation events that continuously appeared in a de novo manner. This iPS reprogramming consisted of stochastic de novo methylation and selection/fixation of methylation in an environment suitable for ESCs. Taken together, random methylation and convergence are driving forces for long-term reprogramming of iPSCs to ESCs.

  7. Efficacy of Intrauterine Device in the Treatment of Intrauterine Adhesions

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    Umme Salma

    2014-01-01

    Full Text Available The primary purpose of this paper is to assess the efficacy of the use of the intrauterine device (IUD as an adjunctive treatment modality, for intrauterine adhesions (IUAs. All eligible literatures were identified by electronic databases including PubMed, Scopus, and Web of Science. Additional relevant articles were identified from citations in these publications. There were 28 studies included for a systematic review. Of these, 5 studies were eligible for meta-analysis and 23 for qualitative assessment only. Twenty-eight studies related to the use of IUDs as ancillary treatment following adhesiolysis were identified. Of these studies, 25 studies at least one of the following methods were carried out as ancillary treatment: Foley catheter, hyaluronic acid gel, hormonal therapy, or amnion graft in addition to the IUD. There was one study that used IUD therapy as a single ancillary treatment. In 2 studies, no adjunctive therapy was used after adhesiolysis. There was a wide range of reported menstrual and fertility outcomes which were associated with the use of IUD combined with other ancillary treatments. At present, the IUD is beneficial in patients with IUA, regardless of stage of adhesions. However, IUD needs to be combined with other ancillary treatments to obtain maximal outcomes, in particular in patients with moderate to severe IUA.

  8. Amniotic membranes as prosthetic material: experimental utilization data of a rat model.

    Science.gov (United States)

    Zachariou, Z

    1997-10-01

    Prosthetic materials are applied for closing big tissue defects, the repair of traumatized organs, or hernias. Because nonabsorbable synthetic materials are rigid, possess a defined and unchangeable size, and foreign body reaction (FBR) may occur, biological materials may be an alternative. In experimental studies in rats the authors implanted the fetal parts of the human amniotic membranes and examined the utilization and FBR induced in a standardized model. In addition amnion (AM) was combined with vicryl-net (VN) for higher implant stability. Fifteen, 30 and 90 days after implantation, macroscopic appearance was examined, and light microscopy and immunohistology testing of the specimens were performed. Adhesions to parenchymal organs and omentum were present irrespective of the side facing the abdominal cavity. AM induced a rapid FBR, which diminished with time. Chorion (CH) and parts of the AM were resorbed within the examined period after infiltration with recipient cells and neovascularisation. The combined implant, AM, and VN showed best results because disadvantages of one material could be compensated for by the advantages of the other. The studies show that AM, in its anatomic definition, combined with VN proves to be a safe and reliable prosthetic material for the use in tissue defects.

  9. The Evolutionary Economics of Embryonic-Sac Fluids in Squamate Reptiles.

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    Bonnet, Xavier; Naulleau, Guy; Shine, Richard

    2017-03-01

    The parchment-shelled eggs of squamate reptiles take up substantial water from the nest environment, enabling the conversion of yolk into neonatal tissue and buffering the embryo against the possibility of subsequent dry weather. During development, increasing amounts of water are stored in the embryonic sacs (i.e., membranes around the embryo: amnion, allantois, and chorion). The evolution of viviparity (prolonged uterine retention of developing embryos) means that embryonic-sac fluid storage now imposes a cost (increased maternal burdening), confers less benefit (because the mother buffers fetal water balance), and introduces a potential conflict among uterine siblings (for access to finite water supplies). Our data on nine species of squamate reptiles and published information on three species show that the embryonic-sac fluids comprise around 33% of neonatal mass in viviparous species versus 94% in full-term eggs of oviparous squamates. Data on parturition in 149 vipers (Vipera aspis, a viviparous species) show that larger offspring store more fluids in their fetal sacs and that an increase in litter size is associated with a decrease in fluid-sac mass per offspring. Overall, the evolutionary transition from oviparity to viviparity may have substantially altered selective forces on offspring packaging and created competition among offspring for access to water reserves during embryonic development.

  10. Multilineage Potential Research of Bovine Amniotic Fluid Mesenchymal Stem Cells

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    Yuhua Gao

    2014-02-01

    Full Text Available The use of amnion and amniotic fluid (AF are abundant sources of mesenchymal stem cells (MSCs that can be harvested at low cost and do not pose ethical conflicts. In human and veterinary research, stem cells derived from these tissues are promising candidates for disease treatment, specifically for their plasticity, their reduced immunogenicity, and high anti-inflammatory potential. This work aimed to obtain and characterize bovine amniotic fluid mesenchymal stem cells (AFMSC. The bovine AF from the amniotic cavity of pregnant gilts in the early stages of gestation (3- and 4-m-old bovine embryos was collected. AFMSCs exhibit a fibroblastic-like morphology only starting from the fourth passage, being heterogeneous during the primary culture. Immunofluorescence results showed that AFMSCs were positive for β-integrin, CD44, CD73 and CD166, but negative for CD34, CD45. Meanwhile, AFMSCs expressed ES cell markers, such as Oct4, and when appropriately induced, are capable of differentiating into ectodermal and mesodermal lineages. This study reinforces the emerging importance of these cells as ideal tools in veterinary medicine; future studies aimed at a deeper evaluation of their immunological properties will allow a better understanding of their role in cellular therapy.

  11. Use of hyperdry amniotic membrane in operations for cleft palate: a study in rats.

    Science.gov (United States)

    Tsuno, Hiroaki; Noguchi, Makoto; Okabe, Motonori; Tomihara, Kei; Yoshida, Toshiko; Nikaido, Toshio

    2015-04-01

    The growth of maxillary bone and the development of dentition are often impaired in patients who have had pushback operations for repair of a cleft palate. There has been considerable discussion about the most suitable technique or material used in such repairs to resolve the problem. Hyperdry amniotic membrane, a new preservable material derived from human amnion, has recently been introduced in several procedures. We have evaluated its use during pushback surgery in animal studies to try to correct the inhibition of growth and development of the maxilla. Mucosal defects were created in 3-week-old rats, and then covered with hyperdry amniotic membrane or not. Healing was assessed by histological and morphological examination at 1 week and 7 weeks postoperatively. In the group treated with hyperdry amniotic membrane, submucosal tissue was reconstructed successfully during the early postoperative period. Lateral palatal growth was not inhibited as much, and medial inclination of the teeth was less, after a period of growth using this material. The results suggest that hyperdry amniotic membrane is a suitable new dressing material for use in the treatment of cleft palate. Copyright © 2015 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  12. Time of initial detection of fetal and extra-fetal structures by ultrasonographic examination in Miniature Schnauzer bitches.

    Science.gov (United States)

    Kim, Bang Sil; Son, Chang Ho

    2007-09-01

    Serial ultrasonographic examinations were performed daily on 9 Miniature Schnauzer bitches from the 15th day of gestation until parturition to determine the time the gestational structures were first detected. The gestational age was timed from the day of ovulation (day 0), which was estimated to occur when the plasma progesterone concentration was >4.0 ng/ml. The gestational length in 9 Miniature Schnauzer bitches was found to be 63.0 +/- 1.7 (range 61-65) days. The initial detection of the fetal and extra-fetal structures were as follows: gestational sac at day 18.0 +/- 0.9 (17-19); zonary placenta in the uterine wall at day 24.9 +/- 1.1 (23-26); yolk sac membrane at day 25.0 +/- 0.9 (24-26); amnionic membrane at day 27.7 +/- 1.0 (26- 29); embryo initial detection at day 22.6 +/- 0.5 (22-23); heartbeat at day 23.4 +/- 0.5 (23-24); fetal movement at day 32.5 +/- 0.8 (32-34); stomach at day 31.2 +/- 1.6 (29-33); urinary bladder at day 32.6 +/- 1.8 (31-35); skeleton at day 34.9 +/- 1.6 (34-38) and kidney at day 42.2 +/- 0.7 (41-43).

  13. Effects of in ovo feeding of carbohydrates and arginine on hatchability, body weight, energy metabolism and perinatal growth in duck embryos and neonates.

    Science.gov (United States)

    Tangara, M; Chen, W; Xu, J; Huang, F R; Peng, J

    2010-10-01

    1. The objective of this study was to test the hypothesis that in ovo feeding of carbohydrates and arginine into the duck amnion may improve the glycogen store and perinatal growth. At 23 d of incubation, fertile eggs were injected with 1·2 ml of sodium chloride (NaCl), sucrose + maltose (CHO), arginine (Arg) or sucrose + maltose + arginine (CHO + Arg), with controls not injected. Body weight, liver and muscle glycogen levels, and hepatic glucose-6-phosphatase activity were determined at 25 d of incubation, at hatch, and at 3 and 7 d posthatch. 2. At hatch and 7 d of age, the body weights were greater in the in ovo-feeding treatments than the controls. Arg and CHO + Arg significantly enhanced liver glycogen level at hatch compared with controls. CHO and CHO + Arg significantly increased muscle glycogen level at 25 d of incubation over controls. CHO and Arg decreased glucose-6-phosphatase at 25 d of incubation, whereas NaCl and CHO + Arg increased glucose-6-phosphatase at hatch relative to controls. 3. In ovo feeding of carbohydrates and arginine at 23 d of incubation may improve glycogen reserves, which may, in turn, provide the energy needed for perinatal growth.

  14. Antioxidative Defense Enzymes in Placenta Protect Placenta and Fetus in Inherited Thrombophilia from Hydrogen Peroxide

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    Jelena Bogdanovic Pristov

    2009-01-01

    Full Text Available Our aim was to investigate the activities of antioxidative defense enzymes in the placenta, fetal blood and amnion fluid in inherited thrombophilia. Thrombophilia was associated with nearly threefold increase of activity (p < 0.001 of the placental catalase (81.1 ± 20.6 U/mg of proteins in controls and 270.0 ± 69.9 U/mg in thrombophilic subjects, glutathione (GSH peroxidase (C: 20.2 ± 10.1 U/mg; T: 60.0 ± 15.5 U/mg, and GSH reductase (C: 28.9 ± 5.6 U/mg; T: 72.7 ± 23.0 U/mg. The placental activities of superoxide dismutating enzymes—MnSOD and CuZnSOD, did not differ in controls and thrombophilia. Likewise, the activities of catalase and SOD in the fetal blood, and the level of ascorbyl radical which represents a marker of oxidative status of amniotic fluid, were similar in controls and thrombophilic subjects. From this we concluded that in thrombophilia, placental tissue is exposed to H2O2-mediated oxidative stress, which could be initiated by pro-thrombic conditions in maternal blood. Increased activity of placental H2O2-removing enzymes protects fetus and mother during pregnancy, but may increase the risk of postpartum thrombosis.

  15. Mesenchymal Stem Cells from Human Amniotic Membrane and Umbilical Cord Can Diminish Immunological Response in an in vitro Allograft Model.

    Science.gov (United States)

    Dabrowski, Filip A; Burdzinska, Anna; Kulesza, Agnieszka; Chlebus, Marcin; Kaleta, Beata; Borysowski, Jan; Zolocinska, Aleksandra; Paczek, Leszek; Wielgos, Miroslaw

    2017-01-01

    Mesenchymal stem cells (MSCs) are gaining rising interest in gynecology and obstetrics. MSCs immunomodulatory properties are suitable enough to reduce perinatal morbidity caused by inflammation in premature neonates. The aim of this study was to evaluate and compare the ability to inhibit allo-activated lymphocytes proliferation by MSCs derived from different sources: amniotic membrane (AM), umbilical cord (UC) and adipose tissue (AT). MSCs were isolated from AM (n = 7) and UC (n = 6) and AT (n = 6) of healthy women. Cells were characterized by flow cytometry and differentiation assay. To evaluate the potential of fetal and adult MSCs to diminish immunological response, mixed lymphocytes reaction (MLR) was performed. Amnion and UC-derived cells displayed typical MSCs characteristics. Addition of MSCs to MLR significantly inhibited the proliferation of stimulated lymphocytes. The effect was observed regardless of the MSCs type used (p < 0.01 in all groups). Comparative analysis revealed no significant differences in this action between tested MSCs types. Additionally, no type of MSCs significantly stimulated allogeneic lymphocytes. The results prove the immunosuppressive capacities of fetal-derived MSCs in vitro. In the future, they may be potentially used to treat premature newborn as well as in immunomodulation in post-transplant therapy. © 2016 S. Karger AG, Basel.

  16. BAKTERI PATOGEN PERIODONTAL PUTATIF PADA IBU HAMIL SEBAGAI PENYEBAB KELAHIRAN BAYI PREMATUR DENGAN BERAT BADAN LAHIR RENDAH

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    Ayuningtyas Herlianti

    2015-08-01

    Full Text Available Preterm low birth weight (PLBW infants are more likely to die during the neonatal period, and PLBW survivors face neurodevelopmental problems, respiratory problems, and congenital anomalies. It is said that the poor periodontal health of the pregnant woman is a potential risk factor of PLBW. This investigation aims to study the oral hygiene and gingival inflammation condition of 2nd and 3rd trimester of pregnancy women and relate to the putative periodontal pathogen bacteriae as the probable cause of PLBW. The plaque index (PII, calculus index (CI, and bleeding on probing (BOP of the subjects of both groups were measured and compared. Maternal infection during pregnancy has been demonstrated to perturb cytokines and hormone-regulated gestation because of the translocation of the infection. The level of gingival crevicular fluid prostaglandin E2 (PG E2, interleukin 1β (IL-1β, IL-6, tumor necrotizing factors α (TNF-α are usually highly correlated with those in amnion fluid of pregnant women without bacterial vaginosis (BV and with intact placental membrane. It is concluded that the periodontal health of pregnant women must be in good condition to prevent the risk of having PLBW still need to be anticipated due to the presence of the inflammation signs.

  17. Amniotic Fluid β2- Microglobulin Measurements

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    Emine Aydın

    2016-02-01

    Full Text Available OBJECTIVE: To determine β2-microglobulin levels in amniotic fluid during the course of second trimester. STUDY DESIGN: One hundred patient’s amniotic fluid β2-microglobulin levels had been evaluated retrospectively (March-October 2009. The most common amniocentesis indication was advanced maternal age (33.3%. Others were; high risk result for triple test (18.5%, high risk result for double test (6.48%, ventriculomegaly (5.55%, obstetric history for fetus with down syndrome (4.62%, the presence of soft markers on ultrasound (13.8%, others (17.8%. Patients average gravida was 2.66 (range: 1-6, parity was 0.75 (range: 0-3, abortion was 0.65 (range: 0-3. RESULTS: All patients were at second trimester and the average gestational week was 17.7 (range 15- 22. Patients were divided into four groups (15th, 16th, 17-18th and 19-20th gestational weeks. We have demonstrated that amniotic fluid β2-microglobulin levels are increased progressively throughout the second trimester. We have specified normal β2-microglobulin values of each gestational week/period in order to be used in clinical practice. CONCLUSION: We have demonstrated that amnionic fluid β2-microglobulin levels are increased progressively throughout the second trimester.

  18. Release of LHRH-activity from human fetal membranes upon exposure to PGE/sub 2/, oxytocin and isoproterenol

    Energy Technology Data Exchange (ETDEWEB)

    Poisner, A.M.; Poisner, R.; Becca, C.R.; Conn, P.M.

    1986-03-01

    The authors have previously reported that superfused chorion laeve (fetal membranes) release LHRH-like immunoreactivity upon exposure to angiotensin II. They have now studied the effects of other agonists on the release of LHRH-activity and something of its chemical nature. Fetal membranes were obtained from placentas delivered by cesarean section, the amnion stripped from the chorion, and the chorion superfused in an Amicon thin-channel device with the maternal surface facing up. The whole device was submerged in a 37 C water bath and perfused with a modified Locke's solution at 0.4 - 1.0 ml/min. LHRH-activity was measured by radioimmunoassay using three different antisera against LHRH. The release of LHRH-activity was stimulated by 6-10 min exposure to PGE/sub 2/, oxytocin, and isoproterenol. Extracts of chorion were studied using gel filtration on Sephacryl S-200 and ultrafiltration with Amicon PM-10 filters. The bulk of the LHRH-activity appeared as a higher molecular weight form (about 70,000 daltons). Since oxytocin has been reported to release PGE/sub 2/ from chorion, it may release LHRH-activity by virtue of liberating endogenous PGE/sub 2/. The chemical nature of the LHRH-activity is presently under investigation.

  19. Amnioinfusion for preterm rupture of membranes.

    Science.gov (United States)

    Hofmeyr, G J

    2000-01-01

    Preterm rupture of membranes places a fetus at risk of cord compression and amnionitis. Amnioinfusion aims to prevent or relieve umbilical cord compression by infusing a solution into the uterine cavity. The objective of this review was to assess the effects of amnioinfusion for preterm rupture of membranes on maternal and perinatal outcomes. The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Randomised trials of amnioinfusion compared to no amnioinfusion in women with preterm rupture of membranes. Eligibility and trial quality were assessed by the reviewer. One trial of 66 women was included. It had some methodological flaws. No significant differences between amnioinfusion and no amnioinfusion were detected for caesarean section (relative risk 0.32, 95% confidence interval 0.07 to 1.40); low Apgar scores (relative risk 0.28, 95% confidence interval 0.03 to 2.33) or neonatal death (relative risk 0.55, 95% confidence interval 0.05 to 5.77). In the amnioinfusion group, the number of severe fetal heart rate decelerations per hour during the first stage of labour were reduced (weighted mean difference -1.20, 95% confidence interval -1.83 to -0.57). These outcomes are consistent with those found in the Cochrane review on amnioinfusion for cord compression. There is not enough evidence concerning the use of amnioinfusion for preterm rupture of membranes.

  20. Efficacy of Tectonic Corneal Patch Graft for Progressive Peripheral Corneal Thinning

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    Cafer Tanrıverdio

    2014-12-01

    Full Text Available Objectives: To report the results of tectonic corneal patch graft (TCPG in patients with progressive peripheral corneal thinning (PCT. Materials and Methods: In this study, we included 8 patients who underwent TCPG for PCT or perforated corneal ulceration at Ankara Training and Research Hospital. Results: We performed TCPG in 7 patients for PCT and in 1 patient for perforated corneal ulceration. Mean age was 57.2±16.7 (38- 82 years. Postoperative follow-up time ranged from 6 to 24 months (mean 13.9±6.7. Possible etiologies leading to progressive PCT were trachoma, infectious corneal ulcer, and rheumatoid arthritis-severe dry eye in 2 patients each. Other 2 patients had a progressive PCT following ocular surgery. One of the patients with infectious corneal ulcer also had a trauma caused by a scissor. Amnion membrane transplantation was performed in 3 patients prior to TCPG. While the anatomic success was achieved in all 8 patients, best-corrected visual acuity (BCVA was 0.1 or better in 4 patients (50%. Postoperative BCVA was better than preoperative BCVA in 6 patients (75%. Local peripheral anterior synechiae developed in two eyes. Conclusion: TCPG is a useful therapeutic option in selected cases of corneal thinning and perforations because it effectively restores the integrity of the globe and allows acceptable visual results. (Turk J Ophthalmol 2014; 44: 440-4

  1. Amniotic Band Syndrome, Perinatal Hospice, and Palliative Care versus Active Management

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    Shadi Rezai

    2016-01-01

    Full Text Available Introduction. Amniotic band syndrome and sequence are a relatively rare condition in which congenital anomalies occur as a result of the adherence and entrapment of fetal parts with coarse fibrous bands of the amniotic membrane. A large percentage of reported cases have an atypical gestational history. The frequency of this obstetric complication is not affected by fetal gender, genetic abnormality, or prenatal infection. Case. A 21-year-old, G1P0 female parturient at 18 weeks and 5 days with a single intrauterine gestation during a routine ultrasound evaluation was noted to have amniotic band sequence. The pregnancy was subsequently complicated by preterm premature rupture of membranes with oligohydramnios, resulting in a surviving neonate scheduled for rehabilitative treatment. Conclusion. Amniotic band syndrome is an uncommon congenital anomaly resulting in multiple disfiguring and disabling manifestations. Several theories are proposed with most involving early rupture of the amnion and entanglement of fetal parts by amniotic bands. This syndrome can be manifested by development of multiple malformations, with the majority of the defects being limb abnormalities of a disorganized nature, as in the case we present. In the absence of a clear etiology of consequential congenital abnormalities, obstetric management guidelines should use shared decision models to focus on the quality of life for the offspring.

  2. Placenta-an alternative source of stem cells

    International Nuclear Information System (INIS)

    Matikainen, Tiina; Laine, Jarmo

    2005-01-01

    The two most promising practical applications of human stem cells are cellular replacement therapies in human disease and toxicological screening of candidate drug molecules. Both require a source of human stem cells that can be isolated, purified, expanded in number and differentiated into the cell type of choice in a controlled manner. Currently, uses of both embryonic and adult stem cells are investigated. While embryonic stem cells are pluripotent and can differentiate into any specialised cell type, their use requires establishment of embryonic stem cell lines using the inner cell mass of an early pre-implantation embryo. As the blastocyst is destroyed during the process, ethical issues need to be carefully considered. The use of embryonic stem cells is also limited by the difficulties in growing large numbers of the cells without inducing spontaneous differentiation, and the problems in controlling directed differentiation of the cells. The use of adult stem cells, typically derived from bone marrow, but also from other tissues, is ethically non-controversial but their differentiation potential is more limited than that of the embryonic stem cells. Since human cord blood, umbilical cord, placenta and amnion are normally discarded at birth, they provide an easily accessible alternative source of stem cells. We review the potential and current status of the use of adult stem cells derived from the placenta or umbilical cord in therapeutic and toxicological applications

  3. Polimorfisme Gen MMP-9, Ekspresi MMP-9, dan Indeks Apoptosis Sel Serviks pada Kehamilan 21–36 Minggu

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    Udin Sabarudin

    2011-12-01

    Full Text Available Over expression and premature activation of matrix metalloproteinase (MMP can lead to degradation of amnion chorionic membrane which clinically called premature rupture of membrane (PROM. Increasing MMP activity caused by matrix metalloproteinase-9 (MMP-9 gene polymorphism (C-1562T will be followed by apoptosis. This study was aimed to find the differences between MMP-9 expression and cervical apoptotic index (AI and also MMP-9 (C-1562T polymorphism on 21–36 weeks of pregnancy with or without PROM. This was case control study and conducted in Dr. Hasan Sadikin Hospital and Bandung Networking Hospitals (May−November 2009. There were no significant correlation between MMP-9 expression and cervical AI in every variable on both groups. Three cases of PROM were found in mothers below 20 years of age. Women with 28−34 weeks of pregnancy had a greater risk for PROM than 21−28 weeks. Pregnant women with body mass index (BMI 19−26, had risk to have PROM. Only one sample that showed a MMP-9 (C-1562T polymorphism in the premature labor with PROM group. It can be concluded that there are no significant correlation between MMP-9 expression and cervical cells AI on both groups as well as MMP-9 (C-1562T polymorphism which can alter MMP-9 expression.

  4. The roles of the cyclo-oxygenases types one and two in prostaglandin synthesis in human fetal membranes at term.

    Science.gov (United States)

    Sawdy, R J; Slater, D M; Dennes, W J; Sullivan, M H; Bennett, P R

    2000-01-01

    The aim of this study was to determine the relative contributions of cyclo-oxygenase (COX) types 1 and 2 to prostaglandin synthesis at term. Fetal membranes were collected from 6 pregnancies after elective caesarean section at term, prior to labour. The presence of COX-1 and COX-2 protein was determined using Western analysis. The relative contributions of the two isoforms of COX to prostaglandin synthesis were determined by incubation of fetal membrane discs with either a COX-2 selective inhibitor, SC236, or a COX-1 selective inhibitor, SC560, and measurement of prostaglandin release during 24 h using enzyme-linked immuno-sorbent assay (ELISA). Both COX-1 and COX-2 protein were demonstrated in amnion and chorion-decidua. The COX-2 selective inhibitor, SC-236, significantly reduced prostaglandin synthesis, both in its COX-2 specific and higher, non-specific concentration ranges. The COX-1 selective inhibitor, SC-560, had no effect upon prostaglandin synthesis in its COX-1 specific concentration range, but did significantly reduce prostaglandin synthesis at higher, non-selective concentrations. Fetal membranes contain both COX-1 and COX-2 at term, but only COX-2 contributes towards prostaglandin synthesis. COX-2 selective NSAI drugs will be as effective as non-selective agents in inhibition of fetal membrane prostaglandin synthesis and may represent a new strategy for tocolysis. Copyright 2000 Harcourt Publishers Ltd.

  5. Creating a placental inflammatory composite index that has a high prognostic relevance to child morbidity.

    Science.gov (United States)

    Chen, Yan; Zou, Lile; Zhao, Yanjun; Wu, Ting; Ye, Jiangfeng; Zhang, Huijuan; Zhang, Jun

    2017-07-01

    Selecting pathologic measures of placental inflammation that affect pregnancy and childhood health is largely empirical. We aimed to systematically select several core inflammation-related placental measures to construct a novel placental inflammatory evaluation criterion with a high prognostic relevance to child morbidity. We used data from the US Collaborative Perinatal Project (1959-1976), a longitudinal birth cohort study that recruited women during pregnancy and followed the children until 7 years of age. Bootstrap resampling, least absolute shrinkage and selection operator, and receiver-operator curve were used to select placental pathologic measures that were closely related to child morbidity to form a placental inflammatory composite index. Twenty-six candidate placental inflammation-related measures were ranked based on their close association with adverse neonatal outcomes. The top five placental measures were: (i) neutrophilic infiltration in umbilical artery; (ii) placental weight-birthweight ratio; (iii) necrosis in decidua capsularis; (iv) bacterial colony in epithelium of amnion; and (v) opacity of membranes and fetal surface. Several composite indexes were constructed. A five-measure composite index that had the highest prognostic relevance was chosen. Compared with subjects without any of the five abnormal measures, those with any lesion ranging from 1 to 5 had a 1.2- to 4.6-fold risk of adverse child outcomes, respectively. Our composite index is simple, evidence-based, and has predictive value for child morbidity. It may be used as a novel placental inflammatory evaluation criterion. © 2017 Japan Society of Obstetrics and Gynecology.

  6. Repairing Fetal Membranes with a Self-adhesive Ultrathin Polymeric Film: Evaluation in Mid-gestational Rabbit Model.

    Science.gov (United States)

    Pensabene, Virginia; Patel, Premal P; Williams, Phillip; Cooper, Trisha L; Kirkbride, Kellye C; Giorgio, Todd D; Tulipan, Noel B

    2015-08-01

    Preterm premature rupture of membranes causes 40% of all preterm births, affecting 150000 women each year in the United States. Prenatal diagnostic procedures and surgical interventions increase incidence of adverse events, leading to iatrogenic membrane rupture after a fetoscopic procedure in 45% of cases. We propose an ultrathin, self-adherent, poly-L-lactic acid patch ("nanofilm") as a reparative wound closure after endoscopic/fetoscopic procedures. These nanofilms are compatible with application in wet conditions and with minimally invasive instrumentation. Ex vivo studies to evaluate the nanofilm were conducted using human chorion-amnion (CA) membranes. A custom-built inflation device was used for mechanical characterization of CA membranes and for assessment of nanofilm adhesion and sealing of membrane defects up to 3 mm in size. These ex vivo tests demonstrated the ability of the nanofilm to seal human CA defects ranging in size from 1 to 3 mm in diameter. In vivo survival studies were conducted in 25 mid-gestational rabbits, defects were created by perforating the uterus and the CA membranes and subsequently using the nanofilm to seal these wounds. These in vivo studies confirmed the successful sealing of defects smaller than 3 mm observed ex vivo. Histological analysis of whole harvested uteri 7 days after surgery showed intact uterine walls in 59% of the nanofilm repaired fetuses, along with increased uterine size and intrauterine development in 63% of the cases. In summary, we have developed an ultrathin, self-adhesive nanofilm for repair of uterine membrane defects.

  7. Setting up a Tissue Bank in India: The Tata Memorial Hospital Experience.

    Science.gov (United States)

    Gajiwala, A L

    2003-01-01

    In India, the procurement of tissues for transplantation is governed by the Transplantation of Human Organs Act, 1994. However, although this law exists, it is primarily applied to organ transplantation and rules and regulations that are specific to tissue banking have yet to be developed.The Tata Memorial Hospital (TMH) Tissue Bank was started in 1988 as part of an International Atomic Energy Agency (IAEA) programme to promote the use of ionising radiation for the sterilisation of biological tissues. It represents the Government of India within this project and was the first such facility in the country. It is registered with the Health Services Maharashtra State and provides lyophilised amnion, dura mater, skin and bone that have been terminally sterilised with exposure to 25 kGy of gamma radiation from a Cobalt 60 source. These are obtained either from cadavers or live donors.To date the TMH Tissue Bank has provided 6328 allografts for use as biological dressings or in various reconstructive procedures.The TMH Tissue Bank has helped initiate a Tissue Bank at the Defence Laboratory (DL), Jodhpur. At present these are the only two Banks in the country using radiation for terminal sterilisation of banked tissues.The availability of safe, clinically useful and cost effective grafts have resulted in changes in surgical treatment with a concomitant increase in demand for grafts and an interest in developing more tissue banks. The availability of donor tissue however, continues to be a major limitation.

  8. Necrotising enterocolitis in preterm infants: epidemiology and antibiotic consumption in the Polish neonatology network neonatal intensive care units in 2009.

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    Jadwiga Wójkowska-Mach

    Full Text Available The aim of this study was to describe the epidemiology of necrotising enterocolitis (NEC, antibiotic consumption and the usefulness of microbiological tests in very low birth weight (VLBW Polish newborns.Prospective surveillance was performed in the year 2009 by local infection control teams. The study covered 910 infants hospitalized in six Polish neonatal intensive care units. Two kinds of indicators were used for the description of antibiotic usage: the duration of treatment (days of treatment, DOTs and the defined daily dose (DDD.NEC incidence was 8.7% and fatality rate was 19%. Chorioamnionitis, late gestational age and low birth weight were identified as risk factors for NEC. Catheterization, mechanical ventilation and other selected procedures were used considerably longer in newborns with NEC than in the remaining neonates. Total usage of antibiotics reached 2.9 DDDs or 1.437 days; the average use of drugs per case of NEC amounted to 0.47 DDD or 23.2 DOTs. The level of antibiotic usage was analysed with correlation to microbiological tests performed and it was non-significantly greater in the group of children with NEC in whom the tests were performed.A high risk of developing NEC is closely associated with VLBW and with inflammation of the amnion during labour. We observed no relationship between the consumption of antibiotics in neonates with NEC and positive results of microbiological testing indicating sepsis accompanying NEC or gut colonization with pathogens.

  9. Identification of Fetal Inflammatory Cells in Eosinophilic/T-cell Chorionic Vasculitis Using Fluorescent In Situ Hybridization.

    Science.gov (United States)

    Katzman, Philip J; Li, LiQiong; Wang, Nancy

    2015-01-01

    Eosinophilic/T-cell chorionic vasculitis (ETCV) is an inflammatory lesion of placental fetal vessels. In contrast to acute chorionic vasculitis, inflammation in ETCV is seen in chorionic vessel walls opposite the amnionic surface. It is not known whether inflammation in ETCV consists of maternal cells from the intervillous space or fetal cells migrating from the vessel. We used fluorescent in situ hybridization (FISH) to differentiate fetal versus maternal cells in ETCV. Placentas with ETCV, previously identified for a published study, were used. Infant sex in each case was identified using the electronic medical record. For male infants, 3-μm sections were cut from archived tissue blocks from placentas involving ETCV and stained with fluorescent X- and Y-chromosome centromeric probes. A consecutive hematoxylin/eosin-stained section was used for correlation. FISH analysis was performed on 400 interphase nuclei at the site of ETCV to determine the proportion of XX, XY, X, and Y cells. Of 31 ETCV cases, 20 were female and 10 were male (1 sex not recorded). Six of 10 cases with male infants had recuts with visible ETCV. In these 6 cases the average percentages (ranges) of XY cells, X-only cells, and Y-only cells in the region of inflammation were 81 (70-90), 11 (6-17), and 8 (2-14), respectively. There was a 2:1 female:male infant ratio in ETCV. Similar to acute chorionic vasculitis, the inflammation in ETCV is of fetal origin. It is still unknown, however, whether the stimulus for ETCV is of fetal or maternal origin.

  10. Comparative gene expression analysis of Dtg, a novel target gene of Dpp signaling pathway in the early Drosophila melanogaster embryo.

    Science.gov (United States)

    Hodar, Christian; Zuñiga, Alejandro; Pulgar, Rodrigo; Travisany, Dante; Chacon, Carlos; Pino, Michael; Maass, Alejandro; Cambiazo, Verónica

    2014-02-10

    In the early Drosophila melanogaster embryo, Dpp, a secreted molecule that belongs to the TGF-β superfamily of growth factors, activates a set of downstream genes to subdivide the dorsal region into amnioserosa and dorsal epidermis. Here, we examined the expression pattern and transcriptional regulation of Dtg, a new target gene of Dpp signaling pathway that is required for proper amnioserosa differentiation. We showed that the expression of Dtg was controlled by Dpp and characterized a 524-bp enhancer that mediated expression in the dorsal midline, as well as, in the differentiated amnioserosa in transgenic reporter embryos. This enhancer contained a highly conserved region of 48-bp in which bioinformatic predictions and in vitro assays identified three Mad binding motifs. Mutational analysis revealed that these three motifs were necessary for proper expression of a reporter gene in transgenic embryos, suggesting that short and highly conserved genomic sequences may be indicative of functional regulatory regions in D. melanogaster genes. Dtg orthologs were not detected in basal lineages of Dipterans, which unlike D. melanogaster develop two extra-embryonic membranes, amnion and serosa, nevertheless Dtg orthologs were identified in the transcriptome of Musca domestica, in which dorsal ectoderm patterning leads to the formation of a single extra-embryonic membrane. These results suggest that Dtg was recruited as a new component of the network that controls dorsal ectoderm patterning in the lineage leading to higher Cyclorrhaphan flies, such as D. melanogaster and M. domestica. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Fate of egg proteins during the development of Columba livia domestica embryo.

    Science.gov (United States)

    Shbailat, Seba Jamal; Aslan, Ibtisam Omar

    2018-01-01

    The transfer of egg white into the yolk and consumption of yolk proteins by the embryo are largely unexplored in the pigeon Columba livia domestica. Here, we investigated the route of egg white transfer as well as the degradation and uptake of yolk proteins by the pigeon embryo. Initially, we tested the electrophoretic patterns of proteins in different egg compartments throughout development. Then, we used lysozyme as a reference protein to follow the egg white transfer, and we measured its activity using Micrococcus lysodeikticus as a substrate. Moreover, we determined the general protease activity during different developmental stages in the yolk using casein. Finally, we examined the expression of aminopeptidase-N (APN) and oligopeptide transporter PepT1 genes in the yolk sac membrane (YSM) from incubation day 8 until day 17. Several electrophoretic bands of presumptive egg white proteins appeared in different egg compartments. Also, lysozyme activity was detected chronologically in the egg compartments. It appeared on day 12 in the amniotic and intestinal fluids and on day 14 in the yolk. Moreover, protease activity in the yolk increased significantly on day 14 and thereafter. APN expression was largest on day 8 and reduced generally afterward, whereas PepT1 expression peaked between days 13 and 15 but then reduced substantially. Our results suggest that the egg white proteins move through the amnion and intestine into the yolk where they undergo degradation by the activated proteases. Furthermore, the YSM appears to have a role in protein consumption, and this role decreases toward hatch. © 2018 Wiley Periodicals, Inc.

  12. Interleukin 18 messenger RNA and proIL-18 protein expression in chorioamniotic membranes from pregnant women with preterm prelabor rupture of membranes.

    Science.gov (United States)

    Polettini, Jossimara; Vieira, Eliane Passarelli; Santos, Mariana Perlati dos; Peraçoli, José Carlos; Witkin, Steven S; da Silva, Márcia Guimarães

    2012-04-01

    To quantify the expression of IL-18 mRNA and protein in the chorioamniotic membranes of pregnant women with PPROM and correlate expression with histological chorioamnionitis. A case control study that included 42 pregnant women not in labor in the following groups: PPROM (n=28) and controls with intact membranes submitted to selective cesarean section at term (n=14). Expression of IL-18 mRNA in chorioamniotic membranes was determined by real-time polymerase chain reaction, and IL-18 protein expression was measured by western blot. Histopathological analyses and immunolocalization of IL-18 by immunohistochemistry were also performed. Analyses were performed using the Mann-Whitney or Fisher's exact tests and the group effect was considered significant if the adjusted p-values were <0.05 and the magnitude of change was greater than 2-fold for mRNA expression. IL-18 mRNA was present in 100% of samples and no difference in expression was observed between term vs. PPROM membranes (fold-change 0.12; p=0.88). In the PPROM group, no difference was observed in IL-18 mRNA regarding gestational age (fold-change 0.11; p=0.42) or the presence of histological chorioamnionitis (fold-change 0.26; p=0.15). ProIL-18 was present in all samples. IL-18 was immunolocalized to amnion, chorion and decidua cells, with intense immunohistochemical staining at the choriodecidual junction. Chorioamniotic membranes are sources of IL-18 mRNA and proIL-18, and their expression is unrelated to PPROM or histological chorioamnionitis. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  13. Topical application of ST266 reduces UV-induced skin damage

    Directory of Open Access Journals (Sweden)

    Guan L

    2017-11-01

    Full Text Available Linna Guan,1 Amanda Suggs,1 Emily Galan,1 Minh Lam,1 Elma D Baron1,2 1Department of Dermatology, Case Western Reserve University, 2Cleveland Veterans Affairs Medical Center, Cleveland, OH, USA Abstract: Ultraviolet radiation (UVR has a significant impact on human skin and is the major environmental factor for skin cancer formation. It is also believed that 80% of the signs of skin aging are attributed to UVR. UVR induces inflammatory changes in the skin via the increase in oxidative stress, DNA damage vascular permeability, and fluctuation in a myriad of cytokines. Acutely, UVR causes skin inflammation and DNA damage, which manifest as sunburn (erythema. ST266 is the secretome of proprietary amnion-derived cells that have been shown to reduce inflammation and accelerate healing of various wounds by promoting migration of keratinocytes and fibroblasts in preclinical animal studies. We hypothesized that ST266 has anti-inflammatory effects that can be used to reduce ultraviolet (UV erythema and markers of inflammation. In this study, we examined the in vivo effects of ST266 on post UV-irradiated skin by measuring erythema, level of cyclobutane pyrimidine dimer (CPD, and expression level of xeroderma pigmentosum, complementation group A (XPA. We demonstrated that ST266 has the potential to reduce the acute effects of UV-induced skin damage when applied immediately after the initial exposure. In addition, ST266 is shown to reduce erythema, increase XPA DNA repair protein, and decrease damaged DNA. Keywords: ST266, photoaging, erythema, CPD, XPA, UV-induced DNA damage

  14. [Quantitative determination of fetal fibronectin in cervical smears: a new marker for evaluating the risk of premature labor].

    Science.gov (United States)

    Hampl, M; Friese, K; Hofmann, I; Melchert, F

    1994-12-01

    There is a well-known correlation between ascending infection and preterm labour. Nevertheless, up to now an exact method to identify patients which are at high risk for preterm labour does not exist. Fetal fibronectin is an extracellular matrix protein, which is produced by fetal membranes. High concentrations are present in amniotic fluid, but not in cervical secretions in uncomplicated pregnancies (only in 3-4%). If there is an inflammatory mediated damage to fetal membranes (amnion/chorion) or a mechanical disruption caused by preterm contractions, fetal fibronectin should be released into the cervix and vagina. A prospective clinical study measuring quantitatively the content of fFN in cervicovaginal secretions in patients with preterm labour (n = 43), preterm rupture of membranes (n = 15) and 20 controls was undertaken. In 16 patients frequent specimen could be obtained over a period of several weeks. 14 of the 34 patients with preterm labour, which could be observed until delivery, delivered before term ( 75 ng/ml (sensitivity: 92.4%). 13 patients were negative for fFN and only one of these patients delivered before term (92% chance of term delivery in absence of fFN in cervicovaginal fluids). In 8 patients with beta-adrenergic treatment fFN was present. All patients delivered before term. The 15 patients with PROM had very high concentrations of fFN with a medium concentration of 967.3 ng/ml. 18 of the 20 control patients were negative for fFN (< 75 ng/ml), 2 had a slightly elevated concentration of 84 and 85 ng/ml.(ABSTRACT TRUNCATED AT 250 WORDS)

  15. Analysis of circulating hem-endothelial marker RNA levels in preterm infants

    Directory of Open Access Journals (Sweden)

    Kuint Jacob

    2009-06-01

    Full Text Available Abstract Background Circulating endothelial cells may serve as novel markers of angiogenesis. These include a subset of hem-endothelial progenitor cells that play a vital role in vascular growth and repair. The presence and clinical implications of circulating RNA levels as an expression for hematopoietic and endothelial-specific markers have not been previously evaluated in preterm infants. This study aims to determine circulating RNA levels of hem-endothelial marker genes in peripheral blood of preterm infants and begin to correlate these findings with prenatal complications. Methods Peripheral blood samples from seventeen preterm neonates were analyzed at three consecutive post-delivery time points (day 3–5, 10–15 and 30. Using quantitative reverse transcription-polymerase chain reaction we studied the expression patterns of previously established hem-endothelial-specific progenitor-associated genes (AC133, Tie-2, Flk-1 (VEGFR2 and Scl/Tal1 in association with characteristics of prematurity and preterm morbidity. Results Circulating Tie-2 and SCL/Tal1 RNA levels displayed an inverse correlation to gestational age (GA. We observed significantly elevated Tie-2 levels in preterm infants born to mothers with amnionitis, and in infants with sustained brain echogenicity on brain sonography. Other markers showed similar expression patterns yet we could not demonstrate statistically significant correlations. Conclusion These preliminary findings suggest that circulating RNA levels especially Tie2 and SCL decline with maturation and might relate to some preterm complication. Further prospective follow up of larger cohorts are required to establish this association.

  16. Synthesis of a smart pH-responsive magnetic nanocomposite as high loading carrier of pharmaceutical agents.

    Science.gov (United States)

    Berah, Razieh; Ghorbani, Mohsen; Moghadamnia, Ali Akbar

    2017-06-01

    To create facile external controlled drug delivery system, a magnetic porous carrier based on Tin oxide nanoparticles was synthesized by an inexpensive and versatile hydrothermal strategy and used for in-vitro process. Magnetic nanocomposites were qualified by Fourier Transform Infrared (FTIR), Scanning Electron Microscopy (SEM), X-Ray Diffraction (XRD), Vibrational Sample Magnetometer (VSM) and Transmission Electron Microscopy (TEM). Results showed that nanoparticles were synthesized successfully with good dispersion of magnetic nanoparticles in cavity, uniform particle size distribution with average size of 65nm and high magnetization of 33.75 emu/mg. Furthermore, the nano-porosity and magnetism allowed high efficiency and remote controlled drug release. In this study, anti-migraine Sumatriptan was used as drug sample and the effect of drug concentration, Fe/Sn ratio and loading time on drug absorption were investigated. The best result was checked for stability at body temperature and different body pH. The sample with drug concentration of 0.25(mg/ml), Fe/Sn=0.22 and loading time of 1.5h had the highest drug efficiency (70%). Finally, in order to simulate the in vivo process for in-vitro step, Amnion was used and drug diffusion rate was measured in different intervals and different pH values. The result illustrated that after 25h, diffusion reached 65% at pH=2 and 56% at pH=7, and then became constant. Based on the above mentioned results, the carrier has an acceptable in vitro yield and therefore could be chosen for future in vivo researches. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Human Decidua-Derived Mesenchymal Cells Are a Promising Source for the Generation and Cell Banking of Human Induced Pluripotent Stem Cells

    Science.gov (United States)

    Shofuda, Tomoko; Kanematsu, Daisuke; Fukusumi, Hayato; Yamamoto, Atsuyo; Bamba, Yohei; Yoshitatsu, Sumiko; Suemizu, Hiroshi; Nakamura, Masato; Sugimoto, Yoshikazu; Furue, Miho Kusuda; Kohara, Arihiro; Akamatsu, Wado; Okada, Yohei; Okano, Hideyuki; Yamasaki, Mami; Kanemura, Yonehiro

    2013-01-01

    Placental tissue is a biomaterial with remarkable potential for use in regenerative medicine. It has a three-layer structure derived from the fetus (amnion and chorion) and the mother (decidua), and it contains huge numbers of cells. Moreover, placental tissue can be collected without any physical danger to the donor and can be matched with a variety of HLA types. The decidua-derived mesenchymal cells (DMCs) are highly proliferative fibroblast-like cells that express a similar pattern of CD antigens as bone marrow-derived mesenchymal cells (BM-MSCs). Here we demonstrated that induced pluripotent stem (iPS) cells could be efficiently generated from DMCs by retroviral transfer of reprogramming factor genes. DMC-hiPS cells showed equivalent characteristics to human embryonic stem cells (hESCs) in colony morphology, global gene expression profile (including human pluripotent stem cell markers), DNA methylation status of the OCT3/4 and NANOG promoters, and ability to differentiate into components of the three germ layers in vitro and in vivo. The RNA expression of XIST and the methylation status of its promoter region suggested that DMC-iPSCs, when maintained undifferentiated and pluripotent, had three distinct states: (1) complete X-chromosome reactivation, (2) one inactive X-chromosome, or (3) an epigenetic aberration. Because DMCs are derived from the maternal portion of the placenta, they can be collected with the full consent of the adult donor and have considerable ethical advantages for cell banking and the subsequent generation of human iPS cells for regenerative applications. PMID:26858858

  18. DNA repair capacity and rate of excision repair in UV-irradiated mammalian cells

    International Nuclear Information System (INIS)

    Inoue, Masao; Takebe, Hiraku.

    1978-01-01

    Repair capacities of five mammalian cell strains were measured by colony-forming ability, HCR of UV-irradiated virus, UDS, pyrimidine dimer excision, and semi-conservative DNA replication. Colony-forming ability of UV-irradiated cells was high for human amnion FL cells and mouse L cells, slightly low for African green monkey CV-1 cells, and extremely low for xeroderma pigmentosum cells. HCR of UV-irradiated Herpes simplex virus was high in CV-1 cells, FL and normal human fibroblast cells, low in both XP and L cells. The amount of UDS was high in FL and normal human fibroblast cells, considerably low in CV-1 cells, and essentially no UDS was observed in XP cells. Rate of UDS after UV-irradiation was slower for CV-1 cells than FL and human fibroblast cells. Rate of the excision of thymine-containing dimers from the acid-insoluble fraction during post-irradiation incubation of the cells was rapid in FL and normal human cells and slow in CV-1 cells, and no excision took place in XP cells. Semi-conservative DNA synthesis was reduced after UV-irradiation in all cell lines, but subsequently recovered in FL, normal human and CV-1 cells. The onset of recovery was 4 h after UV-irradiation for FL and normal human cells, but about 6 h for CV-1 cells. The apparent intermediate repair of CV-1 cells except for HCR may be related to the slow rate of excision repair. ''Patch and cut'' model is more favorable than ''cut and patch'' model to elucidate these results. (auth.)

  19. Photographic-Based Optical Evaluation of Tissues and Biomaterials Used for Corneal Surface Repair: A New Easy-Applied Method.

    Directory of Open Access Journals (Sweden)

    Miguel Gonzalez-Andrades

    Full Text Available Tissues and biomaterials used for corneal surface repair require fulfilling specific optical standards prior to implantation in the patient. However, there is not a feasible evaluation method to be applied in clinical or Good Manufacturing Practice settings. In this study, we describe and assess an innovative easy-applied photographic-based method (PBM for measuring functional optical blurring and transparency in corneal surface grafts.Plastic compressed collagen scaffolds (PCCS and multilayered amniotic membranes (AM samples were optically and histologically evaluated. Transparency and image blurring measures were obtained by PBM, analyzing photographic images of a standardized band pattern taken through the samples. These measures were compared and correlated to those obtained applying the Inverse Adding-Doubling (IAD technique, which is the gold standard method.All the samples used for optical evaluation by PBM or IAD were histological suitable. PCCS samples presented transmittance values higher than 60%, values that increased with increasing wavelength as determined by IAD. The PBM indicated that PCCS had a transparency ratio (TR value of 80.3 ± 2.8%, with a blurring index (BI of 50.6 ± 4.2%. TR and BI obtained from the PBM showed a high correlation (ρ>|0.6| with the diffuse transmittance and the diffuse reflectance, both determined using the IAD (p<0.005. The AM optical properties showed that there was a largely linear relationship between the blurring and the number of amnion layers, with more layers producing greater blurring.This innovative proposed method represents an easy-applied technique for evaluating transparency and blurriness of tissues and biomaterials used for corneal surface repair.

  20. Surgical tissue banking - the role of nuclear technology

    International Nuclear Information System (INIS)

    Hasim Mohamad

    2002-01-01

    Before 1980s amniotic membranes were mainly used clinically either in fresh forms or preserved in glycerol without sterilization. There were problems of graft infections and difficulty in maintaining standard of graft quality and uniformity of the amnion when applied on the recipient patients. Nuclear technology has helped to sterilize these membranes such that they can be processed, dried and then double packed before finally irradiated with gamma rays emitted from the radionuclide cobalt-60. The irradiated amniotic membranes can be kept at room temperature and are readily available for used by doctors without relying on glycerol for preservation or freezers for storage. Similarly allograft bones can be processed and be stored in similar manner otherwise they need to be frozen at -80 to -160 degree C. Radiation technology has benefited many patients in many countries in the Asia Pacific region through sterilization of tissues. Bone such as femoral heads can be processed into radiation sterilized bone chips or grounded bones and has been successfully used for filling up of cavities left by benign bone tumors, bone cysts etc. These materials are safe and cheap and save the recipients in-term of money and a second surgery and on the extreme case, patients may avoid amputation if an irradiated long bone is available. Orthopedic, neuro- and dental surgeons have reported many such successful outcomes. Also bum patients have been shown to benefit from amniotic membranes or porcine skin which were processed and irradiated prior to treatment. In some countries in this region, amniotic membranes have been used as en effective wound dressing for leprosy patients as well. Reports of these cases are to be presented. (Author)

  1. Effects of in ovo feeding of creatine pyruvate on the hatchability, growth performance and energy status in embryos and broiler chickens.

    Science.gov (United States)

    Zhao, M M; Gao, T; Zhang, L; Li, J L; Lv, P A; Yu, L L; Gao, F; Zhou, G H

    2017-10-01

    The effects of in ovo feeding (IOF) of creatine pyruvate (CrPyr) on the hatchability, growth performance and energy status of embryos and broilers (Arbor Acres) were investigated. Five treatments were arranged as non-injected treatment (Control), 0.6 ml physiological saline (0.75%) injected treatment (Saline), and IOF treatments injected with 0.6 ml physiological saline (0.75%) containing 3, 6 or 12 mg CrPyr (CrPyr3, CrPyr6 or CrPyr12) into the amnion per fertile egg on day 17.5 of incubation. After hatching, 80 male chicks from each treatment with similar weight close to the average BW of their pooled group were selected and randomly assigned into eight replicates of 10 chicks each. The results showed that the hatchability was not affected among groups, whereas the hatching weight of broilers in CrPyr12 was significantly higher than the control and saline groups (P0.05). Irrespective of dosage, the concentrations of creatine and phosphocreatine, and activities of creatine kinase in embryos were enhanced in CrPyr treatments at 19 E when compared with the control and saline groups (P<0.05). The activities of glucose-6-phosphatase in liver in CrPyr6 and CrPyr12 treatments were higher than the control and saline groups at 19 E (P<0.05). In conclusion, these results indicated that IOF of CrPyr, especially at the level of 12 mg/egg, could improve energy status of embryos and hatchlings, which was useful for enhancing hatching weight, BW and pectoral muscle weight until the end of the experiments at 21 days post-hatch in broilers.

  2. Endocrine gland-derived endothelial growth factor (EG-VEGF) is a potential novel regulator of human parturition.

    Science.gov (United States)

    Dunand, C; Hoffmann, P; Sapin, V; Blanchon, L; Salomon, A; Sergent, F; Benharouga, M; Sabra, S; Guibourdenche, J; Lye, S J; Feige, J J; Alfaidy, N

    2014-09-01

    EG-VEGF is an angiogenic factor that we identified as a new placental growth factor during human pregnancy. EG-VEGF is also expressed in the mouse fetal membrane (FM) by the end of gestation, suggesting a local role for this protein in the mechanism of parturition. However, injection of EG-VEGF to gravid mice did not induce labor, suggesting a different role for EG-VEGF in parturition. Here, we searched for its role in the FM in relation to human parturition. Human pregnant sera and total FM, chorion, and amnion were collected during the second and third trimesters from preterm no labor, term no labor, and term labor patients. Primary human chorion trophoblast and FM explants cultures were also used. We demonstrate that circulating EG-VEGF increased toward term and significantly decreased at the time of labor. EG-VEGF production was higher in the FM compared to placentas matched for gestational age. Within the FM, the chorion was the main source of EG-VEGF. EG-VEGF receptors, PROKR1 and PROKR2, were differentially expressed within the FM with increased expression toward term and an abrupt decrease with the onset of labor. In chorion trophoblast and FM explants collected from nonlaboring patients, EG-VEGF decreased metalloproteinase-2 and -9 activities and increased PGDH (prostaglandin-metabolizing enzyme) expression. Altogether these data demonstrate that EG-VEGF is a new cytokine that acts locally to ensure FM protection in late pregnancy. Its fine contribution to the initiation of human labor is exhibited by the abrupt decrease in its levels as well as a reduction in its receptors. © 2014 by the Society for the Study of Reproduction, Inc.

  3. Amniotic Fluid Cells Show Higher Pluripotency-Related Gene Expression Than Allantoic Fluid Cells.

    Science.gov (United States)

    Kehl, Debora; Generali, Melanie; Görtz, Sabrina; Geering, Diego; Slamecka, Jaroslav; Hoerstrup, Simon P; Bleul, Ulrich; Weber, Benedikt

    2017-10-01

    Amniotic fluid represents an abundant source of multipotent stem cells, referred as broadly multipotent given their differentiation potential and expression of pluripotency-related genes. However, the origin of this broadly multipotent cellular fraction is not fully understood. Several sources have been proposed so far, including embryonic and extraembryonic tissues. In this regard, the ovine developmental model uniquely allows for direct comparison of fetal fluid-derived cells from two separate fetal fluid cavities, the allantois and the amnion, over the entire duration of gestation. As allantoic fluid mainly collects fetal urine, cells originating from the efferent urinary tract can directly be compared with cells deriving from the extraembryonic amniotic tissues and the fetus. This study shows isolation of cells from the amniotic [ovine amniotic fluid cells (oAFCs)] and allantoic fluid [ovine allantoic fluid cells (oALCs)] in a strictly paired fashion with oAFCs and oALCs derived from the same fetus. Both cell types showed cellular phenotypes comparable to standard mesenchymal stem cells (MSCs), with trilineage differentiation potential, and expression of common ovine MSC markers. However, the expression of MSC markers per single cell was higher in oAFCs as measured by flow cytometry. oAFCs exhibited higher proliferative capacities and showed significantly higher expression of pluripotency-related genes OCT4, STAT3, NANOG, and REX1 by quantitative real-time polymerase chain reaction compared with paired oALCs. No significant decrease of pluripotency-related gene expression was noted over gestation, implying that cells with high differentiation potential may be isolated at the end of pregnancy. In conclusion, this study suggests that cells with highest stem cell characteristics may originate from the fetus itself or the amniotic fetal adnexa rather than from the efferent urinary tract or the allantoic fetal adnexa.

  4. ACR Appropriateness Criteria® Multiple Gestations.

    Science.gov (United States)

    Glanc, Phyllis; Nyberg, David A; Khati, Nadia J; Deshmukh, Sandeep Prakash; Dudiak, Kika M; Henrichsen, Tara Lynn; Poder, Liina; Shipp, Thomas D; Simpson, Lynn; Weber, Therese M; Zelop, Carolyn M

    2017-11-01

    Women with twin or higher-order pregnancies will typically have more ultrasound examinations than women with a singleton pregnancy. Most women will have at minimum a first trimester scan, a nuchal translucency evaluation scan, fetal anatomy scan at 18 to 22 weeks, and one or more scans in the third trimester to evaluate growth. Multiple gestations are at higher risk for preterm delivery, congenital anomalies, fetal growth restriction, placenta previa, vasa previa, and velamentous cord insertion. Chorionicity and amnionicity should be determined as early as possible when a twin pregnancy is identified to permit triage of the monochorionic group into a closer surveillance model. Screening for congenital heart disease is warranted in monochorionic twins because they have an increased rate of congenital cardiac anomalies. In addition, monochorionic twins have a higher risk of developing cardiac abnormalities in later gestation related to right ventricular outflow obstruction, in particular the subgroups with twin-twin transfusion syndrome or selective intrauterine growth restriction. Monochorionic twins have unique complications including twin-to-twin transfusion syndrome, twin embolization syndrome, and acardius, or twin-reversed arterial perfusion sequence. The American College of Radiology Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed annually by a multidisciplinary expert panel. The guideline development and revision include an extensive analysis of current medical literature from peer reviewed journals and the application of well-established methodologies (RAND/UCLA Appropriateness Method and Grading of Recommendations Assessment, Development, and Evaluation or GRADE) to rate the appropriateness of imaging and treatment procedures for specific clinical scenarios. In those instances where evidence is lacking or equivocal, expert opinion may supplement the available evidence to recommend imaging or

  5. Prophylactic amnioinfusion in preganancies complicated by chorioamnionitis: a prospective randomized trial.

    Science.gov (United States)

    Parilla, B V; McDermott, T M

    1998-01-01

    The objective of this article is to prospectively investigate the efficacy of amnioinfusion as a means to reduce febrile morbidity in pregnancies complicated by chorioamnionitis. All laboring patients with a temperature > or =100.1 degrees F were approached for study participation. Exclusion criteria included amnionitis diagnosed at greater than 8 cm dilation, multiple gestation, placental abruption, or a nonreassuring fetal heart rate tracing. Consenting patients were randomized to receive antibiotics (ampicillin or penicillin with gentamicin) and acetaminophen with or without amnioinfusion. All patients received intrauterine pressure catheter placement. For study patients, normal saline at room temperature was infused at 10 mL/min for 60 min, then 3 mL/min until delivery. Postpartum endometritis was defined as a temperature = 100.4 degrees F accompanied by uterine tenderness more than 12 hr after delivery. Statistical analysis was performed using the Student's t-test for continuous data and Chi-square for discrete variables. Thirty-six patients were enrolled, and complete data were available for 34 patients (17 in each group). There were no differences between groups with respect to maternal age, gravidity, race, or gestational age. There were also no differences between groups in duration of rupture of membranes, temperature at randomization, interval from randomization to delivery, cesarean section rate, or umbilical cord arterial pH. The mean temperature at the time of delivery was 99.8+/-0.9 degrees F for the amnioinfusion group versus 100.5+/-1.0 degrees F for the control group (p=0.046). Three of 17 amnioinfusion patients and 3 of 17 control patients had postpartum endometritis. There was 1 neonatal infection in the treatment group and no neonatal infections among the control patients. Prophylactic amnioinfusion was associated with a decline in temperature at the time of delivery. No untoward effects from the amnioinfusion were identified.

  6. Amnioinfusion for umbilical cord compression in labour.

    Science.gov (United States)

    Hofmeyr, G J

    2000-01-01

    Amnioinfusion aims to prevent or relieve umbilical cord compression during labour by infusing a solution into the uterine cavity. The objective of this review was to assess the effects of amnioinfusion on maternal and perinatal outcome for potential or suspected umbilical cord compression or potential amnionitis. The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. Randomised trials of amnioinfusion compared with no amnioinfusion in women with babies at risk of umbilical cord compression; and women at risk of intrauterine infection. Eligibility and trial quality were assessed by the reviewer. Twelve studies were included. Transcervical amnioinfusion for potential or suspected umbilical cord compression was associated with the following reductions: fetal heart rate decelerations (relative risk 0.54, 95% confidence interval 0.43 to 0.68); caesarean section for suspected fetal distress (relative risk 0.35, 95% confidence interval 0.24 to 0.52); neonatal hospital stay greater than 3 days (relative risk 0.40, 95% confidence interval 0. 26 to 0.62); maternal hospital stay greater than 3 days (relative risk 0.46, 95% 0.29 to 0.74). Transabdominal amnioinfusion showed similar results. Transcervical amnioinfusion to prevent infection in women with membranes ruptured for more than 6 hours was associated with a reduction in puerperal infection (relative risk 0.50, 95% confidence interval 0.26 to 0.97). Amnioinfusion appears to reduce the occurrence of variable heart rate decelerations and lower the use of caesarean section. However the studies were done in settings where fetal distress was not confirmed by fetal blood sampling. The results may therefore only be relevant where caesarean sections are commonly done for abnormal fetal heart rate alone. The trials reviewed are too small to address the possibility of rare but serious maternal adverse effects of amnioinfusion.

  7. Quantitative and subcellular localization analysis of the nuclear isoform dUTP pyrophosphatase in alkylating agent-induced cell responses

    International Nuclear Information System (INIS)

    Hu, Xiaolan; Yu, Yingnian; Li, Qian; Wu, Danxiao; Tan, Zhengning; Wang, Cheng; Wang, Jvping; Wu, Meiping

    2011-01-01

    Highlights: → MNNG-induced appearance of DUT-N in the extracellular fluid has cellular specificity. → MNNG alters the subcellular distribution of DUT-N in human cells in different ways. → DUT-N may be a potential biomarker to assess the risk of alkylating agents exposure. -- Abstract: Our previous proteome analysis showed that the nuclear isoform of dUTP pyrophosphatase (DUT-N) was identified in the culture medium of human amnion FL cells after exposure to the alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). These results suggest that DUT-N may be a potential early biomarker to assess the risk of alkylating agents exposure. DUT-N is one of the two isoforms of deoxyuridine triphosphate nucleotidohydrolase (dUTPase). Our current knowledge of DUT-N expression in human cells is very limited. In the current study, we first investigated the appearance of DUT-N in the culture medium of different human cell lines in response to a low concentration of MNNG exposure. We verified that the MNNG-induced appearance of DUT-N in the extracellular environment is cell-specific. Western blot analysis confirmed that the intracellular DUT-N changes responded to MNNG in a concentration-dependent and cell-specific manner. Furthermore, subcellular fraction experiments showed that 0.25 μM MNNG treatment dramatically increased the DUT-N expression levels in the cytoplasmic extracts prepared from both FL and HepG2 cells, increased DUT-N levels in nuclear extracts prepared from HepG2 cells, and decreased DUT-N levels in nuclear extracts from FL cells. Morphological studies using immunofluorescence showed that a low concentration of MNNG could alter the distribution of DUT-N in FL and HepG2 cells in different ways. Taken together, these studies indicate a role of DUT-N in alkylating agent-induced cell responses.

  8. Effect of Intravenous Dexamethasone on Preparing the Cervix and Labor Induction

    Directory of Open Access Journals (Sweden)

    Fatemeh Laloha

    2015-10-01

    Full Text Available The use of corticosteroids is one of the methods put forward for the strengthening and speeding up the process of labor. After identification of glucocorticoid receptors in human amnion, the role of corticosteroids in starting the process of labor has been studied in numerous studies. The purpose of this study was to determine the effect of intravenous Dexamethasone on preparing the cervix and on labor induction. A randomized, clinical, and double – blind trial was conducted on 172 women divided into a control and an experimental group. The inclusion criteria were that they had to be primparous, in or before the 40th week of pregnancy, and with Bishop scores (B.S.s of 4 or lower. The exclusion criteria were diabetes, preeclampsia, macrosomia, twin pregnancy, rupture of the membrane (ROM, breech, and women suffering from background diseases. The B.S.s of the women was measured in charge of the study, and each woman was intravenously injected with eight milligrams of Dexamethasone or eight milligrams of distilled water. Four hours after the injections, the B.S.s of the participants was measured, and they were put under the conditions of labor induction using oxytocin. Information was collected in checklists A and B. The patients were compared with respect to B.S., the time the induction started, the average interval between the start of induction and the beginning of the active phase of childbirth, and the average length of time between the start of the active phase and the second stage of childbirth. The first and five minutes Apgar scores of the two groups of women were compared. The frequencies, the means,  and the standard deviations were calculated using the SPSS – 16 software, and analysis of the results was performed with the Student’s t- test and the chi-square test with PPPP

  9. Intraventricular hemorrhage risk factors in very low birth weight newborns: a case-control study

    Directory of Open Access Journals (Sweden)

    Laura Martins

    2009-09-01

    Full Text Available Objective: Identification of variables that affect the risk of severe intraventricular hemorrhage (IVH in very low birth weight (VLBW newborns. Methods: Analytic case-control study, in a population consisting of all VLBW newborns admitted to the Neonatal Intensive Care Unit of a maternity hospital, between January 2002 and December 2007. The authors considered as cases all VLBW newborns with severe IVH (grade ≥ 3, and control all VLBW newborns without IVH. Independent variables included obstetric, perinatal and neonatal diagnosis and therapy. Bivariate analysis and multivariate logistic regression analysis were performed. Rresults: During this period, of the 864 VLBW newborns admitted to the Neonatal Intensive Care Unit, 9.7% had severe IVH. With bivariate analysis an association between severe IVH, gestational age and birth weight was found. Prenatal care and pre-eclampsia were associated with a decrease in the incidence of severe IVH. Amnionitis, being outborn, vaginal delivery, male gender, intubation in the delivery room, surfactant, hyaline membrane disease, pneumothorax, necrotizing enterocolitis (NEC perforation and oscillatory high frequency ventilation were associated with an increased incidence of severe IVH. By multivariate logistic regression, the variables associated with increased risk of severe IVH were: pneumothorax (OR = 3.8; 95%CI = 1.7-8.3, NEC with perforation (OR = 8.8; 95%CI = 1.7-45.0, vaginal delivery (OR = 2.0; 95%CI = 1.0-4.1 and high frequency ventilation (OR = 4.8; 95%CI = 1.3-17.3. The following were protective of severe IVH: gestational age (OR = 0.61; 95%CI = 0.52-0.72, patent ductus arteriosus treatment with indomethacin (OR = 0.26; 95%CI = 0.11-0.6 and fertility treatment (OR = 0.24; 95%CI = 0.06-0.94. Cconclusion: These data outline the importance of improvement of pre and neonatal care to reduce severe IVH.

  10. Limbal Fibroblasts Maintain Normal Phenotype in 3D RAFT Tissue Equivalents Suggesting Potential for Safe Clinical Use in Treatment of Ocular Surface Failure.

    Science.gov (United States)

    Massie, Isobel; Dale, Sarah B; Daniels, Julie T

    2015-06-01

    Limbal epithelial stem cell deficiency can cause blindness, but transplantation of these cells on a carrier such as human amniotic membrane can restore vision. Unfortunately, clinical graft manufacture using amnion can be inconsistent. Therefore, we have developed an alternative substrate, Real Architecture for 3D Tissue (RAFT), which supports human limbal epithelial cells (hLE) expansion. Epithelial organization is improved when human limbal fibroblasts (hLF) are incorporated into RAFT tissue equivalent (TE). However, hLF have the potential to transdifferentiate into a pro-scarring cell type, which would be incompatible with therapeutic transplantation. The aim of this work was to assess the scarring phenotype of hLF in RAFT TEs in hLE+ and hLE- RAFT TEs and in nonairlifted and airlifted RAFT TEs. Diseased fibroblasts (dFib) isolated from the fibrotic conjunctivae of ocular mucous membrane pemphigoid (Oc-MMP) patients were used as a pro-scarring positive control against which hLF were compared using surrogate scarring parameters: matrix metalloproteinase (MMP) activity, de novo collagen synthesis, α-smooth muscle actin (α-SMA) expression, and transforming growth factor-β (TGF-β) secretion. Normal hLF and dFib maintained different phenotypes in RAFT TE. MMP-2 and -9 activity, de novo collagen synthesis, and α-SMA expression were all increased in dFib cf. normal hLF RAFT TEs, although TGF-β1 secretion did not differ between normal hLF and dFib RAFT TEs. Normal hLF do not progress toward a scarring-like phenotype during culture in RAFT TEs and, therefore, may be safe to include in therapeutic RAFT TE, where they can support hLE, although in vivo work is required to confirm this. dFib RAFT TEs (used in this study as a positive control) may be useful toward the development of an ex vivo disease model of Oc-MMP.

  11. Transport and biodistribution of dendrimers across human fetal membranes: implications for intravaginal administration of dendrimer-drug conjugates.

    Science.gov (United States)

    Menjoge, Anupa R; Navath, Raghavendra S; Asad, Abbas; Kannan, Sujatha; Kim, Chong J; Romero, Roberto; Kannan, Rangaramanujam M

    2010-06-01

    Dendrimers are emerging as promising topical antimicrobial agents, and as targeted nanoscale drug delivery vehicles. Topical intravaginal antimicrobial agents are prescribed to treat the ascending genital infections in pregnant women. The fetal membranes separate the extra-amniotic space and fetus. The purpose of the study is to determine if the dendrimers can be selectively used for local intravaginal application to pregnant women without crossing the membranes into the fetus. In the present study, the transport and permeability of PAMAM (poly (amidoamine)) dendrimers, across human fetal membrane (using a side by side diffusion chamber), and its biodistribution (using immunofluorescence) are evaluated ex-vivo. Transport across human fetal membranes (from the maternal side) was evaluated using Fluorescein (FITC), an established transplacental marker (positive control, size approximately 400 Da) and fluorophore-tagged G(4)-PAMAM dendrimers (approximately 16 kDa). The fluorophore-tagged G(4)-PAMAM dendrimers were synthesized and characterized using (1)H NMR, MALDI TOF MS and HPLC analysis. Transfer was measured across the intact fetal membrane (chorioamnion), and the separated chorion and amnion layers. Over a 5 h period, the dendrimer transport across all the three membranes was less than dendrimer (5.8 x 10(-8) cm(2)/s). The biodistribution showed that the dendrimers were largely present in interstitial spaces in the decidual stromal cells and the chorionic trophoblast cells (in 2.5-4 h) and surprisingly, to a smaller extent internalized in nuclei of trophoblast cells and nuclei and cytoplasm of stromal cells. Passive diffusion and paracellular transport appear to be the major route for dendrimer transport. The overall findings further suggest that entry of drugs conjugated to dendrimers would be restricted across the human fetal membranes when administered topically by intravaginal route, suggesting new ways of selectively delivering therapeutics to the mother

  12. Annexin A2 in amniotic fluid: correlation with histological chorioamnionitis, preterm premature rupture of membranes, and subsequent preterm delivery.

    Science.gov (United States)

    Namba, Fumihiko; Ina, Shihomi; Kitajima, Hiroyuki; Yoshio, Hiroyuki; Mimura, Kazuya; Saito, Shigeru; Yanagihara, Itaru

    2012-01-01

    The aim of this study was to determine whether amniotic fluid levels of annexin A2, a phospholipid-binding protein that is abundant in amnion and regulates fibrin homeostasis, are associated with histological chorioamnionitis, preterm premature rupture of the membranes, and subsequent preterm delivery. Amniotic fluid was obtained from 55 pregnant women with preterm labor and/or preterm premature rupture of the membranes before 32weeks of gestation, and amniotic fluid levels of annexin A2 were measured with a sandwich enzyme-linked immunosorbent assay. Amniotic fluid levels of annexin A2 in patients with histological chorioamnionitis was higher than that in the remainder (P=0.053), whereas amniotic fluid levels of annexin A2 in patients with preterm premature rupture of the membranes was significantly higher than that in the remainder (P=0.002). Amniotic levels of annexin A2 was a fair test (area under receiver-operator characteristic curve=0.679), and amniotic fluid levels of annexin A2>878.2ng/mL had a sensitivity of 68.8%, a specificity of 65.2%, a positive predictive value of 73.3%, and a negative predictive value of 60.0% for predicting delivery within 2weeks after amniotic fluid sampling. Furthermore, the combined use of amniotic fluid cut-off levels of 878.2ng/mL for annexin A2 and 13.3ng/mL for interleukin-8 improved the specificity (91.3%) and the positive predictive value (89.5%). We identified amniotic fluid levels of annexin A2, especially in combination with amniotic fluid levels of interleukin-8, as a novel predictive marker for preterm delivery. © 2011 The Authors. Journal of Obstetrics and Gynaecology Research © 2011 Japan Society of Obstetrics and Gynecology.

  13. Comprehensive two-dimensional gel protein databases offer a global approach to the analysis of human cells: the transformed amnion cells (AMA) master database and its link to genome DNA sequence data

    DEFF Research Database (Denmark)

    Celis, J E; Gesser, B; Rasmussen, H H

    1990-01-01

    , mitochondria, Golgi, ribosomes, intermediate filaments, microfilaments and microtubules), levels in fetal human tissues, partial protein sequences (containing information on 48 human proteins microsequenced so far), cell cycle-regulated proteins, proteins sensitive to interferons alpha, beta, and gamma, heat...

  14. Response of human limbal epithelial cells to wounding on 3D RAFT tissue equivalents: effect of airlifting and human limbal fibroblasts.

    Science.gov (United States)

    Massie, Isobel; Levis, Hannah J; Daniels, Julie T

    2014-10-01

    Limbal epithelial stem cell deficiency can cause blindness but may be treated by human limbal epithelial cell (hLE) transplantation, normally on human amniotic membrane. Clinical outcomes using amnion can be unreliable and so we have developed an alternative tissue equivalent (TE), RAFT (Real Architecture for 3D Tissue), which supports hLE expansion, and stratification when airlifted. Human limbal fibroblasts (hLF) may be incorporated into RAFT TEs, where they support overlying hLE and improve phenotype. However, the impact of neither airlifting nor hLF on hLE function has been investigated. hLE on RAFT TEs (±hLF and airlifting) were wounded using heptanol and re-epithelialisation (fluorescein diacetate staining), and percentage putative stem cell marker p63α and proliferative marker Ki67 expression (wholemount immunohistochemistry), measured. Airlifted, hLF- RAFT TEs were unable to close the wound and p63α expression was 7 ± 0.2% after wounding. Conversely, non-airlifted, hLF- RAFT TEs closed the wound within 9 days and p63α expression was higher at 22 ± 5% (p < 0.01). hLE on both hLF- and hLF+ RAFT TEs (non-airlifted) closed the wound and p63α expression was 26 ± 8% and 36 ± 3% respectively (ns). Ki67 expression by hLE increased from 1.3 ± 0.5% before wounding to 7.89 ± 2.53% post-wounding for hLF- RAFT TEs (p < 0.01), and 0.8 ± 0.08% to 17.68 ± 10.88% for hLF+ RAFT TEs (p < 0.05), suggesting that re-epithelialisation was a result of proliferation. These data suggest that neither airlifting nor hLF are necessarily required to maintain a functional epithelium on RAFT TEs, thus simplifying and shortening the production process. This is important when working towards clinical application of regenerative medicine products. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Correlates of vaginal colonization with group B streptococci among pregnant women

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    Tsering Chomu Dechen

    2010-01-01

    Full Text Available Background: A study of genital colonization by group B streptococcus (GBS was conducted in pregnant women in their third trimester, which is a known risk factor of morbidity and mortality among newborns. Aims: The present study was undertaken to study the prevalence and the correlates of vaginal colonization by GBS among pregnant women. Setting and Design: This observational cross-sectional study was conducted during September 2002 to March 2004 on 524 pregnant women. Materials and Methods: Three high vaginal swabs were obtained from all the pregnant women admitted at term and in preterm labor. Two swabs were used for aerobic culture and the third one for gram staining. The first set of swabs was cultured on 5% Sheep blood agar plates. The second set of swabs were inoculated into Todd-Hewitt broth and then subcultured in 5% Sheep blood agar plates. The main outcome measures were the presence of GBS infection in comparison to the age group, gravida, gestational age, premature rupture of membrane (PROM, preterm labor and association with febrile spells of the present pregnancy. Results: The culture positivity rate of GBS was 4.77% and coexistent organisms isolated were Candida species (36%, Staphylococcus aureus (8% and Enterococcus species (8%. Culture positivity in the age group of 18-25 years was 5.71%, of which 5.74% were in their first pregnancy. The correlation between age group and gravida with GBS culture positivity was statistically insignificant. The culture positivity in <36 weeks of gestational age was 6.93%. This relation was statistically significant. Twenty-eight percent developed PROM. Sixty-four percent of culture positives had preterm labor. Conclusion: GBS infection among pregnant women was significantly correlated with the gestational age, PROM and preterm labor. In pregnancy GBS colonization causes asymptomatic bacteriuria or UTI. It is a well known cause of puerperal infections with amnionitis,endometritis and sepsis being

  16. Multicentre epidemiological survey on the incidence of Streptococcus agalactiae in pregnancy

    Directory of Open Access Journals (Sweden)

    Leonardo Lodolo

    2010-03-01

    Full Text Available ß-hemolytic streptococci group B (GBS cause infections of the urinary tract, endometrium, amnion, and superinfection of wounds, with complications in 2% -4% of cases in pregnant woman.The infection transmitted to newborns is a cause of sepsis and meningitis with high incidence of mortality.A recent report on a multicentric study of the ISS, showed that detection of S. agalactiae in pregnant women in different realities of healthcare, ranging from 3.9% to 19.4%. This survey covers a sample of 29607 women screened in a total of 36852 childbirths, with a positivity of 4183 subjects equal to 14.1%. Thanks to the GBS Study Group of Piemonte and Valle d’Aosta microbiology laboratories, with the support of 42 study centres have been involved. Collection techniques, types of medium, environmental of incubation of the culture, possible enrichment in broth, of the swabs of 28491 women during 2006/2007 have been considered. 24.36% were vaginal swab while 75.64% vagino/rectal. According to data collected at 31 centers with eligible data (22,175 pregnant GBS positivity was 12.7% (CI [Confidence Interval] + / -95%: 13.1% -12.2% with wide variability between individual centers (2% -22.6%. The enrichment broth is used by 6 out of 9 centers with positivity> 15% (M: 20.05%, from 6 to 14 with positivity between 10-15% (M: 12.61%, from none to 8 centers with less than 10% positivity (M: 4.95%.This multicentre survey underlined the preference for vagino/rectal swab, as recommended by the literature. The frequency of GBS positivity of pregnant women on the whole samples is similar to that found in other national surveys, even if with large differences between the different centers.The sensitivity tends to increase with the use of enrichment techniques, whose use is uneven between the centers, perhaps indicating the need for greater uniformity of protocols based on the results of microbiological analysis EBM (Evidence Based Medicine.

  17. Effects of in ovo injection of carbohydrates on embryonic metabolism, hatchability, and subsequent somatic characteristics of broiler hatchlings.

    Science.gov (United States)

    Zhai, W; Gerard, P D; Pulikanti, R; Peebles, E D

    2011-10-01

    The effects of the in ovo injection of different carbohydrate solutions on the internal egg temperature (IT), hatchability, and time of hatch of embryonated Ross × Ross 708 broiler hatching eggs were determined. In addition, the BW, liver weight, yolk sac weight (YSW), and yolk-free BW (YFBW) of the embryos on d 19.5 of incubation and of the chicks on day of hatch were determined. Eggs containing live embryos were injected in the amnion on d 18.5 of incubation using an automated multiple-egg injector. Solution injections delivered 1.2 mL of physiological saline (0.85%) alone or with a supplemental carbohydrate. The following supplemental carbohydrates were separately dissolved in saline at a concentration of 0.3 g/mL: glucose, fructose, sucrose, maltose, and dextrin. Temperature transponders were implanted in the air cells of embryonated and nonembryonated eggs after in ovo injection for the detection of IT at 6, 14, and 22 h after injection. The IT of embryonated eggs was significantly greater than that of nonembryonated eggs at all 3 times after the treatment period. Eggs that were injected with saline with or without supplemental carbohydrates experienced a reduction in IT when compared with control eggs whose shells were perforated without solution delivery, and the decrease in IT was associated with a delay in hatch time. Liver weight was negatively related to YSW and positively related to YFBW, and YSW was negatively related to YFBW. Although the saline and carbohydrate solution injections increased chick BW compared with noninjected controls, chick YFBW was decreased in the maltose- and sucrose-injected groups. In conclusion, the injection of 1.2 mL of saline with or without supplemental carbohydrates lowered embryonic metabolism, as reflected by a lower IT and a delay in time of hatch. However, effects of the different carbohydrate solutions on yolk absorption and tissue deposition in yolk-free embryos varied. These results suggest that lower volumes for

  18. Comparative Characterization of Cells from the Various Compartments of the Human Umbilical Cord Shows that the Wharton's Jelly Compartment Provides the Best Source of Clinically Utilizable Mesenchymal Stem Cells.

    Directory of Open Access Journals (Sweden)

    Arjunan Subramanian

    Full Text Available The human umbilical cord (UC is an attractive source of mesenchymal stem cells (MSCs with unique advantages over other MSC sources. They have been isolated from different compartments of the UC but there has been no rigorous comparison to identify the compartment with the best clinical utility. We compared the histology, fresh and cultured cell numbers, morphology, proliferation, viability, stemness characteristics and differentiation potential of cells from the amnion (AM, subamnion (SA, perivascular (PV, Wharton's jelly (WJ and mixed cord (MC of five UCs. The WJ occupied the largest area in the UC from which 4.61 ± 0.57 x 106 /cm fresh cells could be isolated without culture compared to AM, SA, PV and MC that required culture. The WJ and PV had significantly lesser CD40+ non-stem cell contaminants (26-27% compared to SA, AM and MC (51-70%. Cells from all compartments were proliferative, expressed the typical MSC-CD, HLA, and ESC markers, telomerase, had normal karyotypes and differentiated into adipocyte, chondrocyte and osteocyte lineages. The cells from WJ showed significantly greater CD24+ and CD108+ numbers and fluorescence intensities that discriminate between MSCs and non-stem cell mesenchymal cells, were negative for the fibroblast-specific and activating-proteins (FSP, FAP and showed greater osteogenic and chondrogenic differentiation potential compared to AM, SA, PV and MC. Cells from the WJ offer the best clinical utility as (i they have less non-stem cell contaminants (ii can be generated in large numbers with minimal culture avoiding changes in phenotype, (iii their derivation is quick and easy to standardize, (iv they are rich in stemness characteristics and (v have high differentiation potential. Our results show that when isolating MSCs from the UC, the WJ should be the preferred compartment, and a standardized method of derivation must be used so as to make meaningful comparisons of data between research groups.

  19. Zinc finger proteins and other transcription regulators as response proteins in benzo[a]pyrene exposed cells

    International Nuclear Information System (INIS)

    Gao Zhihua; Jin Jinghua; Yang Jun; Yu Yingnian

    2004-01-01

    Proteomic analysis, which combines two-dimensional electrophoresis (2-DE) and mass spectrometry (MS), is an important approach to screen proteins responsive to specific stimuli. Benzo[a]pyrene (B[a]P), a prototype of polycyclic hydrocarbons (PAHs), is a potent procarcinogen generated from the combustion of fossil fuel and cigarette smoke. To further probe the molecular mechanism of mutagenesis and carcinogenesis, and to find potential molecular markers involved in cellular responses to B[a]P exposure, we performed proteomic analysis of whole cellular proteins in human amnion epithelial cells after B[a]P-treatment. Image visualization and statistical analysis indicated that more than 40 proteins showed significant changes following B[a]P-treatment (P<0.05). Among them, 20 proteins existed only in the control groups, while six were only present in B[a]P-treated cells. In addition, the expression of 10 proteins increased whereas 11 decreased after B[a]P-treatment. These proteins were subjected to in-gel tryptic digestion followed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) analysis. Using peptide mass fingerprinting (PMF) to search the nrNCBI database, we identified 22 proteins. Most of these proteins have unknown functions and have not been previously connected to a response to B[a]P exposure. To further annotate the characteristics of these proteins, GOblet analysis was carried out and results indicated that they were involved in multiple biological processes including regulation of transcription, cell proliferation, cell aging and other processes. However, expression changes were noted in a number of transcription regulators, including eight zinc finger proteins as well as SNF2L1 (SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 1), which is closely linked to the chromatin remodeling process. These data may provide new clues to further understand the implication of

  20. Effects of in ovo injection of carbohydrates on somatic characteristics and liver nutrient profiles of broiler embryos and hatchlings.

    Science.gov (United States)

    Zhai, W; Bennett, L W; Gerard, P D; Pulikanti, R; Peebles, E D

    2011-12-01

    Effects of the in ovo injection of commercial diluent supplemented with dextrin or with dextrin in combination with various other carbohydrates on the somatic characteristics and liver nutrient profiles of Ross × Ross 708 broiler embryos and chicks were investigated. Results include information concerning the gluconeogenic energy status of the liver before and after hatch. Eggs containing live embryos were injected in the amnion on d 18 of incubation using an automated multiple-egg injector for the delivery of the following carbohydrates dissolved in 0.4 mL of commercial diluent: 1) 6.25% glucose and 18.75% dextrin; 2) 6.25% sucrose and 18.75% dextrin; 3) 6.25% maltose and 18.75% dextrin; and 4) 25% dextrin. Also, a noninjected control and a 0.4-mL diluent-injected control were included. Body weight relative to set egg weight on d 19 of incubation (E19) was increased by the injection of all carbohydrate solutions, and on the day of hatch was increased by the injection of diluent, sucrose and dextrin, and maltose and dextrin solutions. Hatchability of the fertilized eggs, residual yolk sac weight, and liver weight were not affected by any injection treatment; however, as compared with the 0.4 mL diluent-injected group, all of the supplementary carbohydrates, except for the glucose and dextrin combination group, increased liver glycogen and glucose concentrations on E19. Furthermore, all carbohydrates, except for the 25% dextrin treatment, decreased liver fat concentration on E19. From E19 to the day of hatch, liver glycogen concentrations dropped dramatically from an average of 3.2 to 0.6%. Despite treatment differences observed on E19 for liver glycogen, glucose, and fat concentrations, these differences were lost by the day of hatch. Nevertheless, liver glycogen and glucose concentrations were positively correlated on the day of hatch. In conclusion, the in ovo injection of various supplemental carbohydrates dissolved in 0.4 mL of commercial diluent altered the

  1. Fetal alcohol exposure and development of the integument

    Directory of Open Access Journals (Sweden)

    Longhurst WD

    2016-05-01

    Full Text Available William D Longhurst,1 Jordan Ernst,2 Larry Burd3 1Center for Emergency Medicine, University Hospitals Case Medical Center, Case Western Reserve University, Cleveland, OH, USA; 2University of North Dakota School of Medicine and Health Sciences, Grand Forks, ND, USA; 3Department of Pediatrics, North Dakota Fetal Alcohol Syndrome Center, University of North Dakota School of Medicine and Health Sciences, Grand Forks, ND, USA Background: The physiology of fetal alcohol exposure changes across gestation. Early in pregnancy placental, fetal, and amniotic fluid concentrations of alcohol exposure are equivalent. Beginning in mid-pregnancy, the maturing fetal epidermis adds keratins which decrease permeability resulting in development of a barrier between fetal circulation and the amniotic fluid. Barrier function development is essential for viability in late pregnancy and in the extra-uterine environment. In this paper we provide a selected review of the effects of barrier function on fetal alcohol exposure. Methods: We utilized a search of PubMed and Google for all years in all languages for MeSH on Demand terms: alcohol drinking, amnion, amniotic fluid, epidermis, ethanol, female, fetal development, fetus, humans, keratins, permeability, and pregnancy. We also reviewed the reference lists of relevant papers and hand-searched reference lists of textbooks for additional references. Results: By 30 gestational weeks, development of barrier function alters the pathophysiology of ethanol dispersion between the fetus and amniotic fluid. Firstly, increases in the effectiveness of barrier function decreases the rate of diffusion of alcohol from fetal circulation across fetal skin into the amniotic fluid. This reduces the volume of alcohol entering the amniotic fluid. Secondly, barrier function increases the duration of fetal exposure by decreasing the rate of alcohol diffusion from amniotic fluid back into fetal circulation. Ethanol is then transported into

  2. Effects of in ovo feeding of cationic amino acids on hatchability, hatch weights, and organ developments in domestic pigeon squabs (Columba livia).

    Science.gov (United States)

    Zhang, X Y; Li, L L; Miao, L P; Zhang, N N; Zou, X T

    2018-01-01

    .8, P < 0.05). In conclusion, cationic amino acids injection into amnion can improve the embryonic development, which may be mediated by the increment of relative organ weight. © 2017 Poultry Science Association Inc.

  3. Perinatal Outcomes Associated With Isolated Velamentous Cord Insertion in Singleton and Twin Pregnancies.

    Science.gov (United States)

    Sinkin, Joshua A; Craig, Wendy Y; Jones, Michael; Pinette, Michael G; Wax, Joseph R

    2018-02-01

    To evaluate perinatal outcomes in singleton and twin pregnancies with pathologically confirmed velamentous cord insertion without vasa previa. This retrospective case-control study included all nonanomalous singleton and twin pregnancies with pathologically confirmed velamentous cord insertion delivered in a single institution between January 1, 2005, and July 1, 2015, and having an ultrasound examination by maternal-fetal medicine. For each case, the next 2 consecutive deliveries matched for gestational age at delivery ± 1 week and, in twins, amnionicity and chorionicity served as controls. Primary outcomes included surgical delivery for a nonreassuring intrapartum fetal heart rate tracing, umbilical arterial cord pH of less than 7.2, 5-minute Apgar score of less than 7, birth weight below the 10th percentile, neonatal intensive care unit admission, fetal or neonatal death, and cord avulsion necessitating manual placental extraction. Outcomes were available for 53 singletons with 103 matched controls and 33 twin pregnancies with 65 matched controls. In singletons, velamentous cord insertion was associated with cord pH of less than 7.2 (odds ratio [OR] 3.5; 95% confidence interval [CI], 1.1-11.2; P = .039), 5-minute Apgar score of less than 7 (OR, 5.3; 95% CI, 0.99-28.1; P = .045), and cord avulsion requiring manual placental extraction (7.5% versus 0%; P = .012). Associations were suggested with increased surgical delivery for a nonreassuring intrapartum fetal heart rate tracing (OR, 2.4; 95% CI, 0.9-6.9; P = .14), birth weight below the 10th percentile (OR, 2.1; 95% CI, 0.8-5.9; P = .21), and fetal or neonatal death (3.8% versus 0%; P = .11). Velamentous cord insertions were also associated with placental abruption in singletons (7.5% versus 0%; P = .013). Among twins, velamentous cord insertion was associated with fetal or neonatal death (9.1% versus 0%; P = .036). Isolated confirmed velamentous cord insertion is associated

  4. Level, causes, and risk factors of stillbirth: a population-based case control study from Chandigarh, India.

    Science.gov (United States)

    Newtonraj, Ariarathinam; Kaur, Manmeet; Gupta, Madhu; Kumar, Rajesh

    2017-11-13

    Globally, India ranks first in the absolute number of stillbirths. Hence, the level, causes, and risk factors of stillbirths were estimated to facilitate designing of prevention strategy. Capture and recapture method was used to identify 301 stillbirths from 1st July 2013 to 31st August 2014 in Chandigarh Union Territory of India. Verbal autopsies (n = 181) were done at household level to identify causes of stillbirths. Risk factors were determined using case-control approach. Women who had a stillbirth in the past 3 months were enrolled as cases (n = 181) and those who had live-birth in same neighbourhood were included as controls (n = 181). Statistical differences in the distribution of characteristics of cases and controls were tested by t test and chi square test respectively for quantitative and categorical variables. In logistic regression models adjusted odds ratios (aOR) and 95% confidence intervals (CIs) were estimated for various risk factors. Stillbirth rate was estimated to be 16/1000 birth. Antepartum causes were more common (68%) than intrapartum causes (32%). Among maternal conditions, hypertension (18.2%) and chorio-amnionitis (13.8%), and among foetal conditions, growth restriction (19.9%) and congenital anomalies (18.8%) were the leading causes. In about half of the stillbirths foetal (48%) and maternal (44.7%) causes were unidentifiable. Risk factors of stillbirths were: higher maternal age (aOR 1.1, 95%CI 1.0-1.2), vaginal delivery (aOR 8.1, 95%CI 2.6-26), induced labour (aOR 2.6, 95%CI 1.5-4.5), green or light brown liquor (aOR 2.0, 95%CI 1.1-3.8), preterm delivery (aOR 6.4, 95%CI 3.7-11) and smaller household size (aOR 1.2, 95% CI 1.1-1.3). Stillbirth rate was high in Chandigarh Union Territory of India. Major causes and risk factors amenable to interventions were infections, hypertension, congenital malformations, foetal growth restriction, pre-maturity and household size. Therefore, better maternity ante-natal and intra-natal care is

  5. Zinc ferrite nanoparticles activate IL-1b, NFKB1, CCL21 and NOS2 signaling to induce mitochondrial dependent intrinsic apoptotic pathway in WISH cells

    Energy Technology Data Exchange (ETDEWEB)

    Saquib, Quaiser; Al-Khedhairy, Abdulaziz A.; Ahmad, Javed; Siddiqui, Maqsood A.; Dwivedi, Sourabh; Khan, Shams T. [Department of Zoology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451 (Saudi Arabia); Chair for DNA Research, Department of Zoology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451 (Saudi Arabia); Musarrat, Javed, E-mail: musarratj1@yahoo.com [Chair for DNA Research, Department of Zoology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451 (Saudi Arabia); Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim University, Aligarh 202002, U.P. (India)

    2013-12-01

    The present study has demonstrated the translocation of zinc ferrite nanoparticles (ZnFe{sub 2}O{sub 4}-NPs) into the cytoplasm of human amnion epithelial (WISH) cells, and the ensuing cytotoxicity and genetic damage. The results suggested that in situ NPs induced oxidative stress, alterations in cellular membrane and DNA strand breaks. The [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) and neutral red uptake (NRU) cytotoxicity assays indicated 64.48 ± 1.6% and 50.73 ± 2.1% reduction in cell viability with 100 μg/ml of ZnFe{sub 2}O{sub 4}-NPs exposure. The treated WISH cells exhibited 1.2-fold higher ROS level with 0.9-fold decline in membrane potential (ΔΨm) and 7.4-fold higher DNA damage after 48 h of ZnFe{sub 2}O{sub 4}-NPs treatment. Real-time PCR (qPCR) analysis of p53, CASP 3 (caspase-3), and bax genes revealed 5.3, 1.6, and 14.9-fold upregulation, and 0.18-fold down regulation of bcl 2 gene vis-à-vis untreated control. RT{sup 2} Profiler™ PCR array data elucidated differential up-regulation of mRNA transcripts of IL-1b, NFKB1, NOS2 and CCL21 genes in the range of 1.5 to 3.7-folds. The flow cytometry based cell cycle analysis suggested the transfer of 15.2 ± 2.1% (p < 0.01) population of ZnFe{sub 2}O{sub 4}-NPs (100 μg/ml) treated cells into apoptotic phase through intrinsic pathway. Over all, the data revealed the potential of ZnFe{sub 2}O{sub 4}-NPs to induce cellular and genetic toxicity in cells of placental origin. Thus, the significant ROS production, reduction in ΔΨm, DNA damage, and activation of genes linked to inflammation, oxidative stress, proliferation, DNA damage and repair could serve as the predictive toxicity and stress markers for ecotoxicological assessment of ZnFe{sub 2}O{sub 4}-NPs induced cellular and genetic damage. - Highlights: • First report on the molecular toxicity of ZnFe{sub 2}O{sub 4}-NPs in cells of placental origin • WISH cells treated with ZnFe{sub 2}O{sub 4}-NPs exhibited cytoplasmic

  6. IGF-I and NEFA concentrations in fetal fluids of term pregnancy dogs.

    Science.gov (United States)

    Meloni, Tea; Comin, Antonella; Rota, Alessandro; Peric, Tanja; Contri, Alberto; Veronesi, Maria Cristina

    2014-06-01

    between amnion and allantois, but in both fetal fluids, higher NEFA levels were detected in samples belonging to small breeds when compared with medium and large. These data strongly indicated that, also in the dog, a relation between fetal fluids IGF-I and NEFA concentrations and breed size exists. Further research is needed to elucidate the possible role of IGF-I and NEFA in the pathologic conditions related to canine fetal growth. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. [Clinical study of 12 cases with obstetric mirror syndrome].

    Science.gov (United States)

    Wu, Lin-lin; Wang, Chen-hong; Li, Zhi-quan

    2012-03-01

    To discuss the clinical features, management, pregnancy outcome and prognosis of obstetric mirror syndrome. The clinical data of 12 cases with obstetric mirror syndrome at Shenzhen Maternity and Child Healthcare Hospital from April 2008 to December 2010 were collected to retrospectively analyze the clinical features, management, pregnancy outcome and prognosis. (1) ETIOLOGY: 12 cases with obstetric mirror syndrome included 9 cases of Bart's hydrops fetalis, 2 cases with fetal complicated congenital cardiac anomalies, and 1 case of unknown etiology. (2) Gestational age at diagnosis and at delivery: gestational age at diagnosis ranged from 28 to 36 weeks [mean (31.5 ± 4.7) weeks], and gestational age at delivery ranged from 28(+3) to 38 weeks [mean (32.9 ± 2.9) weeks]. There were no significant differences between the gestational age at diagnosis and at delivery in consistence with severe preeclampsia group and mild preeclampsia group [(31.8 ± 2.3) weeks vs. (30.9 ± 7.2) weeks, (32.5 ± 2.3) weeks vs. (33.5 ± 3.9) weeks, P > 0.05]. (3) The patients with obstetric mirror syndrome can present a preeclampsia-like syndrome: maternal extremity edema in 12 cases, headache and visual disturbance in 1 case, proteinuria in 11 cases, elevated blood pressure in 5 cases, elevated uric acid in 9 cases, hypoproteinemia in 12 cases, elevated creatinine in 3 case, elevated liver enzyme in 1 case, thrombocytopenia in 2 cases. The major complications included 1 case of HELLP syndrome, acute pulmonary edema, placental abruption, amnionic fluid embolism, DIC respectively, 3 cases of acute kidney failure and 6 cases of postpartum hemorrhage. (4) Sonographic findings: 1) Hydrops fetalis: fetal ultrasound revealed pleural fluid, fetal ascites, skin edema, scalp edema, encephalocolele enlargement, hydropericardium and increased cardio-chest ratio. 2) Placenta megaly: the placental pathological examination revealed edematous and large in 12 cases. Placental thickness was beyond 4 cm in

  8. Membrana amniótica preservada em glicerina no reparo de feridas cutâneas de membros locomotores de eqüinos Equine amniotic membrane preserved in glycerin in reparing the equine limbs cütaneous wounds

    Directory of Open Access Journals (Sweden)

    Valdemir Alves de Oliveira

    1998-12-01

    treated with amniotic membrane, and the other, the control group, treated only with a damp gauze changed every 48 hours. In all the wounds the procedure was to evaluate the measurement of the área, degree of secretion, development of exuberant granulation tissue and the complete time of epitheliazation. Bacteriologic and histopatologic exams were made in a sample of the treated and contrai groups. The biochemical determination of proteins and the identification of fibroblastic growth factor were realized in the amnionic membrane. In all the treated wounds were observed shorter healing time, less formation of exuberant granulation tissue, smaller incidence of pathogenic bacterias and a smaller increase of the wound area.

  9. First birth Caesarean section and subsequent fertility: a population-based study in the USA, 2000-2008.

    Science.gov (United States)

    Kjerulff, K H; Zhu, J; Weisman, C S; Ananth, C V

    2013-12-01

    Is first birth Caesarean delivery associated with a lower likelihood of subsequent childbearing when compared with first birth vaginal delivery? In this study of US women whose first delivery was in 2000, those who had a Caesarean delivery were less likely to have a subsequent live birth than those who delivered vaginally. Some studies have reported lower birth rates subsequent to Caesarean delivery in comparison with vaginal delivery, while other studies have reported no difference. We conducted a retrospective cohort study of 52 498 women who had a first singleton live birth in the State of Pennsylvania, USA in 2000 and were followed to the end of 2008 via Pennsylvania birth certificate records to identify subsequent live births during the 8- to 9-year follow-up period. Birth certificate records of first singleton births were linked to the hospital discharge data for each mother and newborn, and linked to all birth certificate records for each mother's subsequent deliveries which occurred in 2000 to the end of 2008. Poisson regression models were used to evaluate the association between first birth factors and whether or not there was a subsequent live birth during the follow-up period. Over an average of 8.5 years of follow-up, 40.2% of women with a Caesarean first birth did not have a subsequent live birth, compared with 33.1% of women with a vaginal first birth (risk ratio (RR): 1.21, 95% confidence interval (CI): 1.18-1.25). Adjustment for the demographic confounders of maternal age, race, education, marital status and health insurance coverage attenuated the RR to 1.16 (95% CI: 1.13-1.19). Specific pregnancy and childbirth-related complications associated with not having a subsequent live birth included diabetes-related disorders, abnormalities of organs and soft tissues of the pelvis, fetal abnormalities, premature or prolonged rupture of membranes, hypertensive disorders, amnionitis, fetal distress and other maternal health problems. However, adjustment

  10. The epidemiologic evidence linking prenatal and postnatal exposure to endocrine disrupting chemicals with male reproductive disorders: a systematic review and meta-analysis.

    Science.gov (United States)

    Bonde, Jens Peter; Flachs, Esben Meulengracht; Rimborg, Susie; Glazer, Clara Helene; Giwercman, Aleksander; Ramlau-Hansen, Cecilia Høst; Hougaard, Karin Sørig; Høyer, Birgit Bjerre; Hærvig, Katia Keglberg; Petersen, Sesilje Bondo; Rylander, Lars; Specht, Ina Olmer; Toft, Gunnar; Bräuner, Elvira Vaclavik

    2016-12-01

    More than 20 years ago, it was hypothesized that exposure to prenatal and early postnatal environmental xenobiotics with the potential to disrupt endogenous hormone signaling might be on the causal path to cryptorchidism, hypospadias, low sperm count and testicular cancer. Several consensus statements and narrative reviews in recent years have divided the scientific community and have elicited a call for systematic transparent reviews. We aimed to fill this gap in knowledge in the field of male reproductive disorders. The aim of this study was to systematically synthesize published data on the risk of cryptorchidism, hypospadias, low sperm counts and testicular cancer following in utero or infant exposure to chemicals that have been included on the European Commission's list of Category 1 endocrine disrupting chemicals defined as having documented adverse effects due to endocrine disruption in at least one intact organism. A systematic literature search for original peer reviewed papers was performed in the databases PubMed and Embase to identify epidemiological studies reporting associations between the outcomes of interest and exposures documented by biochemical analyses of biospecimens including maternal blood or urine, placenta or fat tissue as well as amnion fluid, cord blood or breast milk; this was followed by meta-analysis of quantitative data. The literature search resulted in 1314 references among which we identified 33 papers(28 study populations) fulfilling the eligibility criteria. These provided 85 risk estimates of links between persistent organic pollutants and rapidly metabolized compounds (phthalates and Bisphenol A) and male reproductive disorders. The overall odds ratio (OR) across all exposures and outcomes was 1.11 (95% CI 0.91-1.35). When assessing four specific chemical subgroups with sufficient data for meta-analysis for all outcomes, we found that exposure to one of the four compounds, p,p'-DDE, was related to an elevated risk: OR 1.35 (95

  11. Selected Abstracts of the 20th National Congress of the Italian Society of Perinatal Medicine (Società Italiana di Medicina Perinatale, SIMP; Catania (Italy; March 22-24, 2018; Session “Perinatal infections”

    Directory of Open Access Journals (Sweden)

    --- Various Authors

    2018-03-01

    Full Text Available 20th National Congress of the Italian Society of Perinatal Medicine (Società Italiana di Medicina Perinatale, SIMP Catania (Italy • March 22nd-24th, 2018SIMP PRESIDENTIrene CetinCONGRESS PRESIDENTSNicola Chianchiano, Angela Motta SCIENTIFIC COMMITTEEMariarosaria Di Tommaso, Gianpaolo Donzelli, Luca Ramenghi----------Guest Editors: Irene Cetin, Nicola Chianchiano, Angela Motta ----------Session “Perinatal infections”ABS 1. ROLE OF ABNORMAL CTG FEATURES IN CLINICAL AND SUBCLINICAL CHORIO­AMNIONITIS • L. Galli, V. Whelehan, A. Archer, A. Dall’Asta, T. Frusca, E. ChandraharanABS 2. HISTOLOGICAL CHORIOAMNIONITIS AT TERM: WHAT CLINICAL VALUE? • A. Pintucci, S. Consonni, L.E. Gioia, P. Colombo, P. Di Lorenzo, I. Vaglio Tessitore, F. Moltrasio, G. Bovo, A. LocatelliABS 3. VALNOCTAMIDE RESCUES CYTO­MEGALO­VIRUS-INDUCED ABNORMAL BRAIN ON­TOGENY AND DEAFNESS • S. Ornaghi, J. Bai, W. Tan, J. Santos-Sacchi, D. Navaratnam, A.N. van den Pol, M.J. Paidas, P. VerganiABS 4. ZIKA TESTING IN PREGNANT AND PREG­NANCY PLANNING WOMEN • G. Liuzzi, C. Castilletti E. Nicastri, F. Vairo, A. Corpolongo E. Lalle, M. Iannetta, L. Bordi, L. Scorzolini, F. Carletti, S. Quartu, F. Colavita, M.R. Capobianchi, G. IppolitoABS 5. MEASLES IS AGAIN A PROBLEM • C.I. Palermo, C.M. Costanzo, R. Russo, M. Di Franco, G. ScaliaABS 6. OUTCOMES ASSOCIATED WITH FETAL PARVOVIRUS B19 INFECTION: A SYSTEMATIC REVIEW AND META-ANALYSIS • D. Buca, F. Bascietto, M. Liberati, D. Murgano, A. Iacovelli, M.E. Flacco, L. Manzoli, A. Familiari, G. Scambia, F. D’AntonioABS 7. HIV INFECTION AMONG MIGRANT PEOPLE: A SINGLE CENTER EXPERIENCE • A. Giudicepietro, L. Sarno, G.M. Maruotti, L.L. Mazzarelli, M.C. De Angelis, A. Sirico, F. Zullo, P. Martinelli, M. SansoneABS 8. FETAL PARVOVIRUS B19 INFECTION • V. Giardini, S. Lazzarin, A. Martinelli, S. Cozzolino, M. Verderio, P. VerganiABS 9. MASTITIS AND BREAST ABSCESSES IN BREASTFEEDING: MILK CULTURE, ANTIBIOTIC TREATMENT AND FOLLOW

  12. [Effects of human amniotic epithelial stem cells-derived exosomes on healing of wound with full-thickness skin defect in rats].

    Science.gov (United States)

    Zhao, B; Wu, G F; Zhang, Y J; Zhang, W; Yang, F F; Xiao, D; Zeng, K X; Shi, J H; Su, L L; Hu, D H

    2017-01-20

    Objective: To investigate the effects of human amniotic epithelial stem cells-derived exosomes on healing of wound with full-thickness skin defect in rats. Methods: (1) Human amniotic epithelial stem cells were isolated from the amnion tissue of 5 full-term pregnant women in Department of Obstetrics of our hospital by the method of trypsin digestion, and their morphology was observed. The third passage of cells were stained with rhodamine-phalloidin for cytoskeleton observation. The third passage of cells were identified with flow cytometry through the detection of expressions of cell surface markers CD29, CD31, CD34, CD90, CD105, SSEA3, SSEA4 and immunity-related marker human leukocyte antigen-D related site (HLA-DR). The third passage of cells were also assessed the ability of adipogenic and osteogenic differentiation. (2) The third passage of human amniotic epithelial stem cells were cultured in DMEM medium supplemented with 10% exosome-free fetal bovine serum. Exosomes were isolated from culture supernatant by the method of ultracentrifugation and represented with scanning electron microscope for morphologic observation. (3) Six adult SD rats were anesthetized, and four 1 cm×1 cm sized wounds with full-thickness skin defect were made on the back of each rat. The wounds on the back of each rat were divided into control group, 25 μg/mL exosomes group, 50 μg/mL exosomes group, and 100 μg/mL exosomes group according to the random number table (with 6 wounds in each group), and a total volume of 100 μL phosphate buffered saline, 25 μg/mL exosomes, 50 μg/mL exosomes, and 100 μg/mL exosomes were evenly injected around the wound through multiple subcutaneous sites, respectively. The wound healing rate was calculated based on measurement on post injury day (PID) 7, 14, and 21. On PID 21, the healed wound tissue of each group was collected and stained with HE to observe and count skin accessories, and the arrangement of collagen fibers was observed with Masson

  13. Estudo por microscopia eletrônica do epitélio de neovaginas confeccionadas com membrana amniótica e pesquisa de receptores para estrogênios Electron microscope study of the epithelium of neovagina constructed with amniotic membrane: estrogen receptor evaluation

    Directory of Open Access Journals (Sweden)

    Mauri José Piazza

    1999-06-01

    cortes histológicos obtidos em biópsias de neovaginas revestidas com membrana amniótica.Purpose: to verify changes in amnion epithelia after neovaginoplasty and the presence of estrogen receptor in this tissue using electron microscopy (TEM. Methods: a group consisting of 33 young patients with vaginal agenesy, most of them amenorreic, either incapable or having difficulty to adequately perform sexual activity, underwent surgical correction of the anomaly by the McIndoe-Banister technique. In each woman, a cavity was created through an opening of the vesicorectal space. A mold made of artificial sponge with a condom recovered by amniotic membrane was introduced into the cavity. After eight days, the mold was removed, leaving a developing neoepithelium from the amniotic membrane. Results: The TEM conducted in samples of tissue obtained from neovaginas showed similar results in all three layers of these epithelia, either in the cytoplasm or the cellular microstructures in comparison with normal women. The intensity of staining of the estrogen receptors on the vaginal neoepithelium of the group of women who underwent the surgical procedure was similar to that of normal women. The estrogen receptors were heterogeneously distributed in the three layers of the tissue in all women. The analysis of the vaginal neoepithelium obtained from amniotic membrane revealed, through the TEM, all the characteristics of a regular vaginal epithelium.

  14. Relação entre a quantidade de AgNORS, atividade proliferativa e o estágio de desenvolvimento placentário em equinos Relationship between the amount of AgNORs, proliferative activity and stage of placental development in horses

    Directory of Open Access Journals (Sweden)

    Ana C.F. Mançanares

    2012-12-01

    silver nitrate and are related to the activity of rRNA synthesis and to the agility and speed of cell proliferation in the tissues studied. The objective of this study was to relate the amount of AgNORs, proliferative activity and stage of pregnancy in horses, using the coloring of Silver Nitrate. The embryonic attachments were collected, fixed in 10% buffered formaldehyde, embedded in paraplast and stained by silver nitrate. The groups were determined according to the gestational age. The amount of the corium NOR found in early pregnancy indicates the onset of cell activity, and in that the pregnancy progresses, the amount of NOR increases, suggesting higher activity and increased synthesis of their importance in maintaining the fetus. Contrary to what occurs in the corium, the quantification of NORs was higher in late pregnancy than in the beginning, suggesting the stabilization of these membranes in late pregnancy. The chorionic girdle and the yolk sac were found in early pregnancy and had lots of NORs, suggesting synthesis function and proliferation in early pregnancy, since their functions is maintenance of the embryo until the complete formation of the true placenta (chorio-allantoic membranes. We conclude that the membranes that develop in a progressive manner in accordance with the growing embryo/fetal (chorion, amnion and allantoic membranes have an increased number of NORs and the membranes that involute after the formation of the embryo/fetus (yolk sac and chorionic girdle have a decrease in number, suggesting a reduction in proliferative activity in these membranes.

  15. Evidence for extended age dependent maternal immunity in infected children: mother to child transmission of HIV infection and potential interventions including sulfatides of the human fetal adnexa and complementary or alternative medicines.

    Science.gov (United States)

    Bhargav, Hemant; Huilgol, Vidya; Metri, Kashinath; Sundell, I Birgitta; Tripathi, Satyam; Ramagouda, Nagaratna; Jadhav, Mahesh; Raghuram, Nagarathna; Ramarao, Nagendra Hongasandra; Koka, Prasad S

    2012-01-01

    The two neighboring southwestern states of India, Karnataka and Maharashtra, have high incidence of HIV/AIDS and are among the six most high prevalence HIV infected states. In Karnataka state, the northern districts of Bagalkot, Belgaum and Bijapur (the three Bs) and in Maharashtra state, the southern districts of Sangli, Satara, and Solapur (the three Ss) are the areas with the highest incidence of HIV/AIDS. We have evaluated the incidence of maternal to child transmission (MTCT) of HIV-1 infection in Belgaum District which is more than 500 kilometers distance by road from the campus in greater Bangalore (Karnataka State). We have obtained the prenatal CD4 counts of HIV infected pregnant mothers. We have also screened the HIV infected children in two orphanages (rehabilitation centres for HIV infected children) in Belgaum District. The clinical conditions of these infected children were assessed for their CD4 counts, anti-retroviral therapy (ART) intake status, outpatient illnesses and body composition. We have observed that there is an influence of the age factor on the CD4 counts of the HIV infected children. Further, in view of the role of our recently found involvement of sulfatide, 3-O- galactosylceramide, in inhibition of HIV-1 replication and enhancement of hematopoiesis which is otherwise inhibited due to such infection, we have discussed the possible role of sulfatides that biologically occur in the fetal adnexa (placentatrophoblasts /amnion/chorion-umbilical cord), in containing HIV infection as a potential safer alternative to the ART regimens currently approved to be clinically practiced. Lastly, we have discussed the complementary and alternative medicine (CAM) therapies such as evidence based yoga and ayurveda as add-on to ART in potential elimination of MTCT of HIV infection. Out of a total of 150 children delivered by HIV infected mothers, 13 children were found to be positive as determined by the dried blood smear (DBS) for virological testing