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Sample records for amniotic fluid stem

  1. Stem cells from amniotic fluid--Potential for regenerative medicine.

    Science.gov (United States)

    Loukogeorgakis, Stavros P; De Coppi, Paolo

    2016-02-01

    Regenerative medicine has recently been established as an emerging field focussing on repair, replacement or regeneration of cells, tissues and whole organs. The significant recent advances in the field have intensified the search for novel sources of stem cells with potential for therapy. Recently, researchers have identified the amniotic fluid as an untapped source of stem cells that are multipotent, possess immunomodulatory properties and do not have the ethical and legal limitations of embryonic stem cells. Stem cells from the amniotic fluid have been shown to differentiate into cell lineages representing all three embryonic germ layers without generating tumours, which make them an ideal candidate for tissue engineering applications. In addition, their ability to engraft in injured organs and modulate immune and repair responses of host tissues suggest that transplantation of such cells may be useful for the treatment of various degenerative and inflammatory diseases affecting major tissues/organs. This review summarises the evidence on amniotic fluid cells over the past 15 years and explores the potential therapeutic applications of amniotic fluid stem cells and amniotic fluid mesenchymal stem cells. Copyright © 2015. Published by Elsevier Ltd.

  2. Amniotic fluid as a source of engraftable stem cells

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    Cesar V Borlongan

    2017-01-01

    Full Text Available The ability of stem cells to differentiate into various lineages has made them powerful tools of regenerative medicine and applicable to multiple human diseases. Of particular interest, amniotic fluid-derived stem cells (AFSC have been characterized to express both adult and embryonic cell markers, indicating them as cells within an intermediate stage between embryonic and adult phenotype. AFSC can differentiate into cells of all three germ layers, including hepatic, myogenic, osteogenic, and neurogenic cell types. Furthermore, AFSC have minimal replicative senescence, retaining the ability to divide effectively for over 250 doublings. These facts indicate that amniotic fluid may exist as a promising donor source of stem cells for the treatment of multiple clinically relevant conditions. Of particular interest is the convenience of harvesting stem cells from the amniotic fluid stem for the treatment of newborns, as well as for banking or cryopreserving purposes to be used at a later date. Importantly, the promise of amniotic fluid as a source of stem cells merits ongoing research into their potential therapeutic applications. This paper is a review article. Referred literature in this paper has been listed in the references section. The datasets supporting the conclusions of this article are available online by searching various databases, including PubMed. Some original points in this article come from the laboratory practice in our research center and the authors' experiences.

  3. 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.

  4. Amniotic Fluid Stem Cells: A New Era in Regenerative Medicine.

    Science.gov (United States)

    Srivastava, Mala; Ahlawat, Neha; Srivastava, Ankita

    2018-02-01

    Regenerative medicine has become an emerging field which focuses on repair, replacement or regeneration of cells, tissues and the entire organs. The regeneration may occur in patient's own body by using their system as a bioreactor, e.g., cell therapy that involves transplantation of stem cells capable of proliferating, differentiating and replacing damaged host cells. As the field of regenerative medicine advances, and sources of stem cells has been intensified. Though embryonic and adult tissues can be used for isolation of pluripotent stem cells, the amniotic fluid (AF) has been proposed as an alternative source of stem cells for tissue regeneration. AF cells could be banked and used for either allogeneic or autologous transplantation.

  5. Concise Review: Amniotic Fluid Stem Cells: The Known, the Unknown, and Potential Regenerative Medicine Applications.

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    Loukogeorgakis, Stavros P; De Coppi, Paolo

    2017-07-01

    The amniotic fluid has been identified as an untapped source of cells with broad potential, which possess immunomodulatory properties and do not have the ethical and legal limitations of embryonic stem cells. CD117(c-Kit)+ cells selected from amniotic fluid have been shown to differentiate into cell lineages representing all three embryonic germ layers without generating tumors, making them ideal candidates for regenerative medicine applications. Moreover, their ability to engraft in injured organs and modulate immune and repair responses of host tissues, suggest that transplantation of such cells may be useful for the treatment of various degenerative and inflammatory diseases. Although significant questions remain regarding the origin, heterogeneous phenotype, and expansion potential of amniotic fluid stem cells, evidence to date supports their potential role as a valuable stem cell source for the field of regenerative medicine. Stem Cells 2017;35:1663-1673. © 2016 AlphaMed Press.

  6. Development of a novel method for amniotic fluid stem cell storage.

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    Zavatti, Manuela; Beretti, Francesca; Casciaro, Francesca; Comitini, Giuseppina; Franchi, Fabrizia; Barbieri, Veronica; Bertoni, Laura; De Pol, Anto; La Sala, Giovanni B; Maraldi, Tullia

    2017-08-01

    Current procedures for collection of human amniotic fluid stem cells (hAFSCs) indicate that cells cultured in a flask for 2 weeks can then be used for research. However, hAFSCs can be retrieved directly from a small amount of amniotic fluid that can be obtained at the time of diagnostic amniocentesis. The aim of this study was to determine whether direct freezing of amniotic fluid cells is able to maintain or improve the potential of a sub-population of stem cells. We compared the potential of the hAFSCs regarding timing of freezing, cells obtained directly from amniotic fluid aspiration (D samples) and cells cultured in a flask before freezing (C samples). Colony-forming-unit ability, proliferation, morphology, stemness-related marker expression, senescence, apoptosis and differentiation potential of C and D samples were compared. hAFSCs isolated from D samples expressed mesenchymal stem cells markers until later passages, had a good proliferation rate and exhibited differentiation capacity similar to hAFSCs of C samples. Interestingly, direct freezing induced a higher concentration of cells positive for pluripotency stem cell markers, without teratoma formation in vivo. This study suggests that minimal processing may be adequate for the banking of amniotic fluid cells, avoiding in vitro passages before the storage and exposure to high oxygen concentration, which affect stem cell properties. This technique might be a cost-effective and reasonable approach to the process of Good Manufacturing Process accreditation for stem-cell banks. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  7. Characteristics of human amniotic fluid mesenchymal stem cells and their tropism to human ovarian cancer.

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    Liru Li

    Full Text Available The mesenchymal stem cells (MSCs derived from amniotic fluid (AF have become an attractive stem cells source for cell-based therapy because they can be harvested at low cost and avoid ethical disputes. In human research, stem cells derived from AF gradually became a hot research direction for disease treatment, specifically for their plasticity, their reduced immunogenicity and their tumor tropism regardless of the tumor size, location and source. Our work aimed to obtain and characterize human amniotic fluid mesenchymal stem cells (AFMSCs and detect their ovarian cancer tropsim in nude mice model. Ten milliliters of twenty independent amniotic fluid samples were collected from 16-20 week pregnant women who underwent amniocentesis for fetal genetic determination in routine prenatal diagnosis in the first affiliated hospital of Harbin medical university. We successfully isolated the AFMSCs from thirteen of twenty amniotic fluid samples. AFMSCs presented a fibroblastic-like morphology during the culture. Flow cytometry analyses showed that the cells were positive for specific stem cell markers CD73,CD90, CD105, CD166 and HLA-ABC (MHC class I, but negative for CD 45,CD40, CD34, CD14 and HLA-DR (MHC class II. RT-PCR results showed that the AFMSCs expressed stem cell marker OCT4. AFMSCs could differentiate into bone cells, fat cells and chondrocytes under certain conditions. AFMSCs had the high motility to migrate to ovarian cancer site but didn't have the tumorigenicity. This study enhances the possibility of AFMSCs as drug carrier in human cell-based therapy. Meanwhile, the research emphasis in the future can also put in targeting therapy of ovarian cancer.

  8. Characteristics of human amniotic fluid mesenchymal stem cells and their tropism to human ovarian cancer.

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    Li, Liru; Wang, Dejun; Zhou, Jun; Cheng, Yan; Liang, Tian; Zhang, Guangmei

    2015-01-01

    The mesenchymal stem cells (MSCs) derived from amniotic fluid (AF) have become an attractive stem cells source for cell-based therapy because they can be harvested at low cost and avoid ethical disputes. In human research, stem cells derived from AF gradually became a hot research direction for disease treatment, specifically for their plasticity, their reduced immunogenicity and their tumor tropism regardless of the tumor size, location and source. Our work aimed to obtain and characterize human amniotic fluid mesenchymal stem cells (AFMSCs) and detect their ovarian cancer tropsim in nude mice model. Ten milliliters of twenty independent amniotic fluid samples were collected from 16-20 week pregnant women who underwent amniocentesis for fetal genetic determination in routine prenatal diagnosis in the first affiliated hospital of Harbin medical university. We successfully isolated the AFMSCs from thirteen of twenty amniotic fluid samples. AFMSCs presented a fibroblastic-like morphology during the culture. Flow cytometry analyses showed that the cells were positive for specific stem cell markers CD73,CD90, CD105, CD166 and HLA-ABC (MHC class I), but negative for CD 45,CD40, CD34, CD14 and HLA-DR (MHC class II). RT-PCR results showed that the AFMSCs expressed stem cell marker OCT4. AFMSCs could differentiate into bone cells, fat cells and chondrocytes under certain conditions. AFMSCs had the high motility to migrate to ovarian cancer site but didn't have the tumorigenicity. This study enhances the possibility of AFMSCs as drug carrier in human cell-based therapy. Meanwhile, the research emphasis in the future can also put in targeting therapy of ovarian cancer.

  9. Nuclear Nox4 Role in Stemness Power of Human Amniotic Fluid Stem Cells

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    Tullia Maraldi

    2015-01-01

    Full Text Available Human amniotic fluid stem cells (AFSC are an attractive source for cell therapy due to their multilineage differentiation potential and accessibility advantages. However the clinical application of human stem cells largely depends on their capacity to expand in vitro, since there is an extensive donor-to-donor heterogeneity. Reactive oxygen species (ROS and cellular oxidative stress are involved in many physiological and pathophysiological processes of stem cells, including pluripotency, proliferation, differentiation, and stress resistance. The mode of action of ROS is also dependent on the localization of their target molecules. Thus, the modifications induced by ROS can be separated depending on the cellular compartments they affect. NAD(PH oxidase family, particularly Nox4, has been known to produce ROS in the nucleus. In the present study we show that Nox4 nuclear expression (nNox4 depends on the donor and it correlates with the expression of transcription factors involved in stemness regulation, such as Oct4, SSEA-4, and Sox2. Moreover nNox4 is linked with the nuclear localization of redox sensitive transcription factors, as Nrf2 and NF-κB, and with the differentiation potential. Taken together, these results suggest that nNox4 regulation may have important effects in stem cell capability through modulation of transcription factors and DNA damage.

  10. The potential of mesenchymal stem cells derived from amniotic membrane and amniotic fluid for neuronal regenerative therapy.

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    Kim, Eun Young; Lee, Kyung-Bon; Kim, Min Kyu

    2014-03-01

    The mesenchymal stem cells (MSCs), which are derived from the mesoderm, are considered as a readily available source for tissue engineering. They have multipotent differentiation capacity and can be differentiated into various cell types. Many studies have demonstrated that the MSCs identified from amniotic membrane (AM-MSCs) and amniotic fluid (AF-MSCs) are shows advantages for many reasons, including the possibility of noninvasive isolation, multipotency, self-renewal, low immunogenicity, anti-inflammatory and nontumorigenicity properties, and minimal ethical problem. The AF-MSCs and AM-MSCs may be appropriate sources of mesenchymal stem cells for regenerative medicine, as an alternative to embryonic stem cells (ESCs). Recently, regenerative treatments such as tissue engineering and cell transplantation have shown potential in clinical applications for degenerative diseases. Therefore, amnion and MSCs derived from amnion can be applied to cell therapy in neuro-degeneration diseases. In this review, we will describe the potential of AM-MSCs and AF-MSCs, with particular focus on cures for neuronal degenerative diseases.

  11. A microfluidic device for separation of amniotic fluid mesenchymal stem cells utilizing louver-array structures.

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    Wu, Huei-Wen; Lin, Xi-Zhang; Hwang, Shiaw-Min; Lee, Gwo-Bin

    2009-12-01

    Human mesenchymal stem cells can differentiate into multiple lineages for cell therapy and, therefore, have attracted considerable research interest recently. This study presents a new microfluidic device for bead and cell separation utilizing a combination of T-junction focusing and tilted louver-like structures. For the first time, a microfluidic device is used for continuous separation of amniotic stem cells from amniotic fluids. An experimental separation efficiency as high as 82.8% for amniotic fluid mesenchymal stem cells is achieved. Furthermore, a two-step separation process is performed to improve the separation efficiency to 97.1%. These results are based on characterization experiments that show that this microfluidic chip is capable of separating beads with diameters of 5, 10, 20, and 40 microm by adjusting the volume-flow-rate ratio between the flows in the main and side channels of the T-junction focusing structure. An optimal volume-flow-rate ratio of 0.5 can lead to high separation efficiencies of 87.8% and 85.7% for 5-microm and 10-microm beads, respectively, in a one-step separation process. The development of this microfluidic chip may be promising for future research into stem cells and for cell therapy.

  12. Amniotic fluid water dynamics

    NARCIS (Netherlands)

    Beall, M. H.; van den Wijngaard, J. P. H. M.; van Gemert, M. J. C.; Ross, M. G.

    2007-01-01

    Water arrives in the mammalian gestation from the maternal circulation across the placenta. It then circulates between the fetal water compartments, including the fetal body compartments, the placenta and the amniotic fluid. Amniotic fluid is created by the flow of fluid from the fetal lung and

  13. Autologous Transplantation of Amniotic Fluid-Derived Mesenchymal Stem Cells into Sheep Fetuses.

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    Shaw, S W Steven; Bollini, Sveva; Nader, Khalil Abi; Gastaldello, Annalisa; Mehta, Vedanta; Filppi, Elisa; Cananzi, Mara; Gaspar, H Bobby; Qasim, Waseem; De Coppi, Paolo; David, Anna L

    2016-03-01

    Long-term engraftment and phenotype correction has been difficult to achieve in humans after in utero stem cell transplantation mainly because of allogeneic rejection. Autologous cells could be obtained during gestation from the amniotic fluid with minimal risk for the fetus and the mother. Using a sheep model, we explored the possibility of using amniotic fluid mesenchymal stem cells (AFMSCs) for autologous in utero stem cell/gene therapy. We collected amniotic fluid (AF) under ultrasound-guided amniocentesis in early gestation pregnant sheep ( n = 9, 58 days of gestation, term = 145 days). AFMSCs were isolated and expanded in all sampled fetal sheep. Those cells were transduced using an HIV vector encoding enhanced green fluorescent protein (GFP) with 63.2% (range 38.3-96.2%) transduction efficiency rate. After expansion, transduced AFMSCs were injected into the peritoneal cavity of each donor fetal sheep at 76 days under ultrasound guidance. One ewe miscarried twin fetuses after amniocentesis. Intraperitoneal injection was successful in the remaining 7 fetal sheep giving a 78% survival for the full procedure. Tissues were sampled at postmortem examination 2 weeks later. PCR analysis detected GFP-positive cells in fetal tissues including liver, heart, placenta, membrane, umbilical cord, adrenal gland, and muscle. GFP protein was detected in these tissues by Western blotting and further confirmed by cytofluorimetric and immunofluorescence analyses. This is the first demonstration of autologous stem cell transplantation in the fetus using AFMSCs. Autologous cells derived from AF showed widespread organ migration and could offer an alternative way to ameliorate prenatal congenital disease.

  14. Amniotic Fluid Stem Cells: A Novel Source for Modeling of Human Genetic Diseases

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    Ivana Antonucci

    2016-04-01

    Full Text Available In recent years, great interest has been devoted to the use of Induced Pluripotent Stem cells (iPS for modeling of human genetic diseases, due to the possibility of reprogramming somatic cells of affected patients into pluripotent cells, enabling differentiation into several cell types, and allowing investigations into the molecular mechanisms of the disease. However, the protocol of iPS generation still suffers from technical limitations, showing low efficiency, being expensive and time consuming. Amniotic Fluid Stem cells (AFS represent a potential alternative novel source of stem cells for modeling of human genetic diseases. In fact, by means of prenatal diagnosis, a number of fetuses affected by chromosomal or Mendelian diseases can be identified, and the amniotic fluid collected for genetic testing can be used, after diagnosis, for the isolation, culture and differentiation of AFS cells. This can provide a useful stem cell model for the investigation of the molecular basis of the diagnosed disease without the necessity of producing iPS, since AFS cells show some features of pluripotency and are able to differentiate in cells derived from all three germ layers “in vitro”. In this article, we describe the potential benefits provided by using AFS cells in the modeling of human genetic diseases.

  15. Amniotic fluid inflammatory cytokines

    DEFF Research Database (Denmark)

    Abdallah, Morsi; Larsen, Nanna; Grove, Jakob

    2013-01-01

    The aim of the study was to analyze cytokine profiles in amniotic fluid (AF) samples of children developing autism spectrum disorders (ASD) and controls, adjusting for maternal autoimmune disorders and maternal infections during pregnancy.......The aim of the study was to analyze cytokine profiles in amniotic fluid (AF) samples of children developing autism spectrum disorders (ASD) and controls, adjusting for maternal autoimmune disorders and maternal infections during pregnancy....

  16. The effect of meconium exposure on the expression and differentiation of amniotic fluid mesenchymal stem cells.

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    Jensen, T J; Shui, J E; Finck, C M

    2017-01-01

    The goal of this study was to determine if exposure to meconium would alter the phenotype of amniotic fluid mesenchymal stem cells (AF-MSCs) and the ability of these cells to be differentiated into distal airway type cells. Meconium was collected, lyophilized and resuspended in PBS at 3 different concentrations (high, medium, and low). AF-MSCs were cultured in the presence of this meconium suspension for 8 hours and then analyzed for changes in gene expression. Additionally, AF-MSCs exposed to meconium were differentiated for 14 days using modified small airway growth medium (mSAGM) and gene expression was determined. As a spontaneous differentiation control, meconium exposed AF-MSCs were cultured in amniotic fluid stem cell medium (AF medium). After 8 hours of exposure in culture, AF-MSCs had increased expression of distal airway genes aquaporin 5 (AQP5) and surfactant protein c (SPC) when cultured in AF medium containing meconium. These gene expression levels were similar to that of AF-MSCs that were differentiated in mSAGM for 14 days. Furthermore, there was an up regulation of pluripotency genes NANOG and OCT4 in response to low meconium concentration for 8 hours. Following 14 days of culture in mSAGM, there was an upregulation of TTF1, SPC and AQP5 expression in the control, as well as in the low and medium meconium exposed groups indicating that these cells were still able to be differentiated. High meconium concentration did, however, appear to influence the level of distal airway gene expression after 14 days in mSAGM. After 14 days in AF medium, there was significant downregulation in pluripotency and mesenchymal markers as well as distal airway gene expression in all groups. The phenotype of AF-MSCs is modulated by meconium exposure; however, the cells were still able to differentiate into distal airway gene and protein expression. This result supports the hypothesis that progenitor cells exist in the amniotic fluid and the presence of meconium may affect

  17. Stem cells isolated from human dental pulp and amniotic fluid improve skeletal muscle histopathology in mdx/SCID mice

    OpenAIRE

    Pisciotta, Alessandra; Riccio, Massimo; Carnevale, Gianluca; Lu, Aiping; De Biasi, Sara; Gibellini, Lara; La Sala, Giovanni B.; Bruzzesi, Giacomo; Ferrari, Adriano; Huard, Johnny; De Pol, Anto

    2015-01-01

    Introduction Duchenne muscular dystrophy (DMD), caused by a lack of the functional structural protein dystrophin, leads to severe muscle degeneration where the patients are typically wheelchair-bound and die in their mid-twenties from cardiac or respiratory failure or both. The aim of this study was to investigate the potential of human dental pulp stem cells (hDPSCs) and human amniotic fluid stem cells (hAFSCs) to differentiate toward a skeletal myogenic lineage using several different proto...

  18. Human amniotic fluid stem cell injection therapy for urethral sphincter regeneration in an animal model

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    Kim Bum

    2012-08-01

    Full Text Available Abstract Background Stem cell injection therapies have been proposed to overcome the limited efficacy and adverse reactions of bulking agents. However, most have significant limitations, including painful procurement, requirement for anesthesia, donor site infection and a frequently low cell yield. Recently, human amniotic fluid stem cells (hAFSCs have been proposed as an ideal cell therapy source. In this study, we investigated whether periurethral injection of hAFSCs can restore urethral sphincter competency in a mouse model. Methods Amniotic fluids were collected and harvested cells were analyzed for stem cell characteristics and in vitro myogenic differentiation potency. Mice underwent bilateral pudendal nerve transection to generate a stress urinary incontinence (SUI model and received either periurethral injection of hAFSCs, periurethral injection of Plasma-Lyte (control group, or underwent a sham (normal control group. For in vivo cell tracking, cells were labeled with silica-coated magnetic nanoparticles containing rhodamine B isothiocyanate (MNPs@SiO2 (RITC and were injected into the urethral sphincter region (n = 9. Signals were detected by optical imaging. Leak point pressure and closing pressure were recorded serially after injection. Tumorigenicity of hAFSCs was evaluated by implanting hAFSCs into the subcapsular space of the kidney, followed two weeks later by retrieval and histologic analysis. Results Flow activated cell sorting showed that hAFSCs expressed mesenchymal stem cell (MSC markers, but no hematopoietic stem cell markers. Induction of myogenic differentiation in the hAFSCs resulted in expression of PAX7 and MYOD at Day 3, and DYSTROPHIN at Day 7. The nanoparticle-labeled hAFSCs could be tracked in vivo with optical imaging for up to 10 days after injection. Four weeks after injection, the mean LPP and CP were significantly increased in the hAFSC-injected group compared with the control group. Nerve regeneration and

  19. Amniotic-fluid-derived mesenchymal stem cells overexpressing interleukin-1 receptor antagonist improve fulminant hepatic failure.

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    Yu-Bao Zheng

    Full Text Available Uncontrolled hepatic immunoactivation is regarded as the primary pathological mechanism of fulminant hepatic failure (FHF. The major acute-phase mediators associated with FHF, including IL-1β, IL-6, and TNF-α, impair the regeneration of liver cells and stem cell grafts. Amniotic-fluid-derived mesenchymal stem cells (AF-MSCs have the capacity, under specific conditions, to differentiate into hepatocytes. Interleukin-1-receptor antagonist (IL-1Ra plays an anti-inflammatory and anti-apoptotic role in acute and chronic inflammation, and has been used in many experimental and clinical applications. In the present study, we implanted IL-1Ra-expressing AF-MSCs into injured liver via the portal vein, using D-galactosamine-induced FHF in a rat model. IL-1Ra expression, hepatic injury, liver regeneration, cytokines (IL-1β, IL-6, and animal survival were assessed after cell transplantation. Our results showed that AF-MSCs over-expressing IL-1Ra prevented liver failure and reduced mortality in rats with FHF. These animals also exhibited improved liver function and increased survival rates after injection with these cells. Using green fluorescent protein as a marker, we demonstrated that the engrafted cells and their progeny were incorporated into injured livers and produced albumin. This study suggests that AF-MSCs genetically modified to over-express IL-1Ra can be implanted into the injured liver to provide a novel therapeutic approach to the treatment of FHF.

  20. An update clinical application of amniotic fluid-derived stem cells (AFSCs) in cancer cell therapy and tissue engineering.

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    Gholizadeh-Ghaleh Aziz, Shiva; Fathi, Ezzatollah; Rahmati-Yamchi, Mohammad; Akbarzadeh, Abolfazl; Fardyazar, Zahra; Pashaiasl, Maryam

    2017-06-01

    Recent studies have elucidated that cell-based therapies are promising for cancer treatments. The human amniotic fluid stem (AFS) cells are advantageous cells for such therapeutic schemes that can be innately changed to express therapeutic proteins. HAFSCs display a natural tropism to cancer cells in vivo. They can be useful in cancer cells targeting. Moreover, they are easily available from surplus diagnostic samples during pregnancy and less ethical and legal concern are associated with the collection and application than other putative cells are subjected. This review will designate representatives of amniotic fluid and stem cell derived from amniotic fluid. For this propose, we collect state of human AFS cells data applicable in cancer therapy by dividing this approach into two main classes (nonengineered and engineered based approaches). Our study shows the advantage of AFS cells over other putative cells types in terms differentiation ability to a wide range of cells by potential and effective use in preclinical studies for a variety of diseases. This study has shown the elasticity of human AFS cells and their favorable potential as a multipotent cell source for regenerative stem cell therapy and capable of giving rise to multiple lineages including such as osteoblasts and adipocyte.

  1. Calcitonin-Induced Effects on Amniotic Fluid-Derived Mesenchymal Stem Cells

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    Caterina Morabito

    2015-05-01

    Full Text Available Background/Aims: Mesenchymal stem cells from human amniotic fluid (huAFMSCs can differentiate into multiple lineages and are not tumorigenic after transplantation, making them good candidates for therapeutic purposes. The aim was to determine the effects of calcitonin on these huAFMSCs during osteogenic differentiation, in terms of the physiological role of calcitonin in bone homeostasis. Methods: For huAFMSCs cultured under different conditions, we assayed: expression of the calcitonin receptor, using immunolabelling techniques; proliferation and osteogenesis, using colorimetric and enzymatic assays; intracellular Ca2+ and cAMP levels, using videomicroscopy and spectrophotometry. Results: The calcitonin receptor was expressed in proliferating and osteo-differentiated huAFMSCs. Calcitonin triggered intracellular Ca2+ increases and cAMP production. Its presence in cell medium also induced dose-dependent inhibitory effects on proliferation and increased osteogenic differentiation of huAFMSCs, as also indicated by enhancement of specific markers and alkaline phosphatase activity. Conclusions: These data show that huAFMSCs represent a potential osteogenic model to study in-vitro cell responses to calcitonin (and other members of the calcitonin family. This leads the way to the opening of new lines of research that will add new insight both in cell therapies and in the pharmacological use of these molecules.

  2. [Comparative studies on different cryopreservation protocols of human amniotic fluid-derived mesenchymal stem cells].

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    Wang, Yiru; Bai, Jing; Chen, Jie; Liu, Lifeng; Wang, Yu

    2012-02-01

    To isolate and culture human amniotic fluid-derived mesenchymal stem cells (HAFMSCs), to investigate a better cryopreservation protocol of HAFMSCs and to observe the bio-characteristics and the multi-potential of HAFMSCs after cryopreservation for the further fundamental researches and clinical applications. HAFMSCs were isolated from the amniotic fluid of pregnant women during the second trimester by the improved two-step method. HAFMSCs were cryopreserved with different cryopreservation protocols (containing different contents of FBS and DMSO at cryoprotectant) in liquid nitrogen for 12 weeks. The bio-characteristics of the HAFMSCs after cryopreservation were analyzed. The growth characteristics were observed by MTT method and the growth curves were drawn. The surface antigens of HAFMSCs were detected using flow cytometry, including CD29, CD34, CD44, CD45, CD73, and CD90. The adipogenic and osteogenic differentiation abilities of HAFMSCs were observed. The mRNA levels of Oct-4 and Nanog of the HAFMSCs were compared between before and after cryopreservation. At 12 weeks after cryopreservation, different protocols had different effects on the cell viability; the better formula of cryoprotectant was 50% DMEM, 40% FBS, and 10% DMSO. After cryopreservation, the cells proliferated rapidly and the growth curves showed "S" shape, which was the same as the cells before cryopreservation. Phenotype showed that HAFMSCs were positive for the surface markers CD29, CD44, CD73, and CD90, and negative for CD34 and CD45. After 21 days of adipogenic differentiation, the lipid droplets were observed by oil red O staining. After 21 days of osteogenic differentiation, the calcium mineralization were verified by von Kossa staining. There was no significant difference (P > 0.05) in the mRNA levels of Oct-4 and Nanog between before and after cryopreservation. HAFMSCs have rapid proliferation and multi-potential in vitro. The cells have high viabilities and no changes of the bio

  3. Upregulation of Nanog and Sox-2 genes following ectopic expression of Oct-4 in amniotic fluid mesenchymal stem cells.

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    Wang, Kai-Hung; Kao, An-Pei; Chang, Chia-Cheng; Lin, Ta-Chin; Kuo, Tsung-Cheng

    2015-01-01

    Octamer-binding transcription factor 4 (Oct-4), an important gene regulating stem cell pluripotency, is well-known for its ability to reprogram somatic cells in vitro, either alone or in concert with other factors. The aim of this study was to assess the effect of ectopic expression of Oct human amniotic fluid stem cells. We developed a novel method for isolation of putative human amniotic fluid-derived multipotent stem cells. These cells showing mesenchymal stem cell phenotypes (human amniotic fluid-derived mesenchymal stem cells, hAFMSCs) were transfected with a plasmid carrying genes for Oct-4 and the green fluorescent protein (GFP). The stably transfected cells, hAFMSCs-Oct4/GFP, were selected by using G418 and found to express the GFP reporter gene under the control of Oct-4 promoter. We found that hAFMSCs developed by our method possess very high self-renewal ability (about 78 cumulative population doublings) and multilineage differentiation potency. Significantly, the hAFMSCs-Oct4/GFP cells showed enhanced expression of the three major pluripotency genes Oct-4, Nanog, and Sox-2, and increased colony-forming ability and growth rate compared with the parental hAFMSCs. We demonstrated that the ectopic expression of Oct-4 gene in hAFMSCs with high self-renewal ability could upregulate Nanog and Sox-2 gene expression and enhance cell growth rate and colony-forming efficiency. Therefore, the ectopic expression of Oct-4 could be a strategy to develop pluripotency in hAFMSCs for clinical applications. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

  4. Direct Reprogramming of Human Amniotic Fluid Stem Cells by OCT4 and Application in Repairing of Cerebral Ischemia Damage.

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    Qin, Mingde; Chen, Ruihua; Li, Hong; Liang, Hansi; Xue, Qun; Li, Fang; Chen, Ying; Zhang, Xueguang

    2016-01-01

    Amniotic fluid stem cells (AFSCs) are a type of fetal stem cell whose stemness encompasses both embryonic and adult stem cells, suggesting that they may be easily and efficiently reprogrammed into induced pluripotent stem cells (iPSCs). To further simplify the reprogramming process, the creation of AFSC-derived iPSCs using a single factor is desirable. Here we report the generation of one-factor human AFSC-iPSCs (AiPSCs) from human AFSCs by ectopic expression of the transcription factor OCT4. Just like human embryonic stem cells, AiPSCs exhibited similar epigenetic status, global gene expression profiles, teratoma formation and in vitro & in vivo pluripotency. Our results indicate that the OCT4 is necessary and sufficient to directly reprogram human AFSCs into pluripotent AiPSCs. Moreover, reflecting the similar memory characteristics of AFSCs and neural stem cells, we show that AiPSC membrane-derived vesicles (MVs) repair cerebral ischemia damage. We anticipate that the successful generation of one-factor AiPSCs will facilitate the creation of patient-specific pluripotent stem cells without the need for transgenic expression of oncogenes. Moreover, MVs from tissue-specific AiPSCs have potential in tissue repair, representing a novel application of iPSCs.

  5. Prenatally engineered autologous amniotic fluid stem cell-based heart valves in the fetal circulation.

    Science.gov (United States)

    Weber, Benedikt; Emmert, Maximilian Y; Behr, Luc; Schoenauer, Roman; Brokopp, Chad; Drögemüller, Cord; Modregger, Peter; Stampanoni, Marco; Vats, Divya; Rudin, Markus; Bürzle, Wilfried; Farine, Marc; Mazza, Edoardo; Frauenfelder, Thomas; Zannettino, Andrew C; Zünd, Gregor; Kretschmar, Oliver; Falk, Volkmar; Hoerstrup, Simon P

    2012-06-01

    Prenatal heart valve interventions aiming at the early and systematic correction of congenital cardiac malformations represent a promising treatment option in maternal-fetal care. However, definite fetal valve replacements require growing implants adaptive to fetal and postnatal development. The presented study investigates the fetal implantation of prenatally engineered living autologous cell-based heart valves. Autologous amniotic fluid cells (AFCs) were isolated from pregnant sheep between 122 and 128 days of gestation via transuterine sonographic sampling. Stented trileaflet heart valves were fabricated from biodegradable PGA-P4HB composite matrices (n = 9) and seeded with AFCs in vitro. Within the same intervention, tissue engineered heart valves (TEHVs) and unseeded controls were implanted orthotopically into the pulmonary position using an in-utero closed-heart hybrid approach. The transapical valve deployments were successful in all animals with acute survival of 77.8% of fetuses. TEHV in-vivo functionality was assessed using echocardiography as well as angiography. Fetuses were harvested up to 1 week after implantation representing a birth-relevant gestational age. TEHVs showed in vivo functionality with intact valvular integrity and absence of thrombus formation. The presented approach may serve as an experimental basis for future human prenatal cardiac interventions using fully biodegradable autologous cell-based living materials. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. [Amniotic fluid, amniocentesis, postmaturity].

    Science.gov (United States)

    Almazán Díaz, M

    1979-01-01

    The exchange among the elements of amniotic liquid is a dinamic process which keeps stable the concentration of their components, showing a variable relation with the maternal plasma in the differents stages of gestation. Thus the formations of the kidneys modifies the amniotic liquid with the urine excretion and then, factors as swallowing; the secretions of the skin and bronchi are factors which are implicated in its production. The amniocentesis is important in the study of high risk fetus, particulary with a prognostic value in the pregnancy sensitized to the Rh factor. The non cultivated celle give us information about the sexual chromatine and enzymes, to get a quick diagnosis, but this has to be confirmed with a culture cells for a final diagnosis, especially in diseases caused by inborn errors of metabolism.

  7. Amniotic fluid stem cells with low γ-interferon response showed behavioral improvement in Parkinsonism rat model.

    Directory of Open Access Journals (Sweden)

    Yu-Jen Chang

    Full Text Available Amniotic fluid stem cells (AFSCs are multipotent stem cells that may be used in transplantation medicine. In this study, AFSCs established from amniocentesis were characterized on the basis of surface marker expression and differentiation potential. To further investigate the properties of AFSCs for translational applications, we examined the cell surface expression of human leukocyte antigens (HLA of these cells and estimated the therapeutic effect of AFSCs in parkinsonian rats. The expression profiles of HLA-II and transcription factors were compared between AFSCs and bone marrow-derived mesenchymal stem cells (BMMSCs following treatment with γ-IFN. We found that stimulation of AFSCs with γ-IFN prompted only a slight increase in the expression of HLA-Ia and HLA-E, and the rare HLA-II expression could also be observed in most AFSCs samples. Consequently, the expression of CIITA and RFX5 was weakly induced by γ-IFN stimulation of AFSCs compared to that of BMMSCs. In the transplantation test, Sprague Dawley rats with 6-hydroxydopamine lesioning of the substantia nigra were used as a parkinsonian-animal model. Following the negative γ-IFN response AFSCs injection, apomorphine-induced rotation was reduced by 75% in AFSCs engrafted parkinsonian rats but was increased by 53% in the control group after 12-weeks post-transplantation. The implanted AFSCs were viable, and were able to migrate into the brain's circuitry and express specific proteins of dopamine neurons, such as tyrosine hydroxylase and dopamine transporter. In conclusion, the relative insensitivity AFSCs to γ-IFN implies that AFSCs might have immune-tolerance in γ-IFN inflammatory conditions. Furthermore, the effective improvement of AFSCs transplantation for apomorphine-induced rotation paves the way for the clinical application in parkinsonian therapy.

  8. Stem cells isolated from human dental pulp and amniotic fluid improve skeletal muscle histopathology in mdx/SCID mice.

    Science.gov (United States)

    Pisciotta, Alessandra; Riccio, Massimo; Carnevale, Gianluca; Lu, Aiping; De Biasi, Sara; Gibellini, Lara; La Sala, Giovanni B; Bruzzesi, Giacomo; Ferrari, Adriano; Huard, Johnny; De Pol, Anto

    2015-08-28

    Duchenne muscular dystrophy (DMD), caused by a lack of the functional structural protein dystrophin, leads to severe muscle degeneration where the patients are typically wheelchair-bound and die in their mid-twenties from cardiac or respiratory failure or both. The aim of this study was to investigate the potential of human dental pulp stem cells (hDPSCs) and human amniotic fluid stem cells (hAFSCs) to differentiate toward a skeletal myogenic lineage using several different protocols in order to determine the optimal conditions for achieving myogenic commitment and to subsequently evaluate their contribution in the improvement of the pathological features associated with dystrophic skeletal muscle when intramuscularly injected into mdx/SCID mice, an immune-compromised animal model of DMD. Human DPSCs and AFSCs were differentiated toward myogenic lineage in vitro through the direct co-culture with a myogenic cell line (C2C12 cells) and through a preliminary demethylation treatment with 5-Aza-2'-deoxycytidine (5-Aza), respectively. The commitment and differentiation of both hDPSCs and hAFSCs were evaluated by immunofluorescence and Western blot analysis. Subsequently, hDPSCs and hAFSCs, preliminarily demethylated and pre-differentiated toward a myogenic lineage for 2 weeks, were injected into the dystrophic gastrocnemius muscles of mdx/SCID mice. After 1, 2, and 4 weeks, the gastrocnemius muscles were taken for immunofluorescence and histological analyses. Both populations of cells engrafted within the host muscle of mdx/SCID mice and through a paracrine effect promoted angiogenesis and reduced fibrosis, which eventually led to an improvement of the histopathology of the dystrophic muscle. This study shows that hAFSCs and hDPSCs represent potential sources of stem cells for translational strategies to improve the histopathology and potentially alleviate the muscle weakness in patients with DMD.

  9. Minimal modulation of the host immune response to SIS matrix implants by mesenchymal stem cells from the amniotic fluid.

    Science.gov (United States)

    Lesage, F; Pranpanus, S; Bosisio, F M; Jacobs, M; Ospitalieri, S; Toelen, J; Deprest, J

    2017-12-01

    Surgical restoration of soft tissue defects often requires implantable devices. The clinical outcome of the surgery is determined by the properties inherent to the used matrix. Mesenchymal stem cells (MSC) modulate the immune processes after in vivo transplantation and their addition to matrices is associated with constructive remodeling. Herein we evaluate the potential of MSC derived from the amniotic fluid (AF-MSC), an interesting MSC source for cell therapeutic applications in the perinatal period, for immune modulation when added to a biomaterial. We implant cell free small intestinal submucosa (SIS) or SIS seeded with AF-MSC at a density of 1 × 10 5 /cm 2 subcutaneously at the abdominal wall in immune competent rats. The host immune response is evaluated at 3, 7 and 14 days postoperatively. The matrix-specific or cellular characteristics are not altered after 24 h of in vitro co-culture of SIS with AF-MSC. The host immune response was not different between animals implanted with cell free or AF-MSC-seeded SIS in terms of cellular infiltration, vascularity, macrophage polarization or scaffold replacement. Profiling the mRNA expression level of inflammatory cytokines at the matrix interface shows a significant reduction in the expression of the pro-inflammatory marker Tnf-α and a trend towards lower iNos expression upon AF-MSC-seeding of the SIS matrix. Anti-inflammatory marker expression does not alter upon cell seeding of matrix implants. We conclude that SIS is a suitable substrate for in vitro culture of AF-MSC and fibroblasts. AF-MSC addition to SIS does not significantly modulate the host immune response after subcutaneous implantation in rats.

  10. Improved neurological outcome by intramuscular injection of human amniotic fluid derived stem cells in a muscle denervation model.

    Directory of Open Access Journals (Sweden)

    Chun-Jung Chen

    Full Text Available The skeletal muscle develops various degrees of atrophy and metabolic dysfunction following nerve injury. Neurotrophic factors are essential for muscle regeneration. Human amniotic fluid derived stem cells (AFS have the potential to secrete various neurotrophic factors necessary for nerve regeneration. In the present study, we assess the outcome of neurological function by intramuscular injection of AFS in a muscle denervation and nerve anastomosis model.Seventy two Sprague-Dawley rats weighing 200-250 gm were enrolled in this study. Muscle denervation model was conducted by transverse resection of a sciatic nerve with the proximal end sutured into the gluteal muscle. The nerve anastomosis model was performed by transverse resection of the sciatic nerve followed by four stitches reconnection. These animals were allocated to three groups: control, electrical muscle stimulation, and AFS groups.NT-3 (Neurotrophin 3, BDNF (Brain derived neurotrophic factor, CNTF (Ciliary neurotrophic factor, and GDNF (Glia cell line derived neurotrophic factor were highly expressed in AFS cells and supernatant of culture medium. Intra-muscular injection of AFS exerted significant expression of several neurotrophic factors over the distal end of nerve and denervated muscle. AFS caused high expression of Bcl-2 in denervated muscle with a reciprocal decrease of Bad and Bax. AFS preserved the muscle morphology with high expression of desmin and acetylcholine receptors. Up to two months, AFS produced significant improvement in electrophysiological study and neurological functions such as SFI (sciatic nerve function index and Catwalk gait analysis. There was also significant preservation of the number of anterior horn cells and increased nerve myelination as well as muscle morphology.Intramuscular injection of AFS can protect muscle apoptosis and likely does so through the secretion of various neurotrophic factors. This protection furthermore improves the nerve

  11. Amniotic fluid stem cells inhibit the progression of bleomycin-induced pulmonary fibrosis via CCL2 modulation in bronchoalveolar lavage.

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    Orquidea Garcia

    Full Text Available The potential for amniotic fluid stem cell (AFSC treatment to inhibit the progression of fibrotic lung injury has not been described. We have previously demonstrated that AFSC can attenuate both acute and chronic-fibrotic kidney injury through modification of the cytokine environment. Fibrotic lung injury, such as in Idiopathic Pulmonary Fibrosis (IPF, is mediated through pro-fibrotic and pro-inflammatory cytokine activity. Thus, we hypothesized that AFSC treatment might inhibit the progression of bleomycin-induced pulmonary fibrosis through cytokine modulation. In particular, we aimed to investigate the effect of AFSC treatment on the modulation of the pro-fibrotic cytokine CCL2, which is increased in human IPF patients and is correlated with poor prognoses, advanced disease states and worse fibrotic outcomes. The impacts of intravenous murine AFSC given at acute (day 0 or chronic (day 14 intervention time-points after bleomycin injury were analyzed at either day 3 or day 28 post-injury. Murine AFSC treatment at either day 0 or day 14 post-bleomycin injury significantly inhibited collagen deposition and preserved pulmonary function. CCL2 expression increased in bleomycin-injured bronchoalveolar lavage (BAL, but significantly decreased following AFSC treatment at either day 0 or at day 14. AFSC were observed to localize within fibrotic lesions in the lung, showing preferential targeting of AFSC to the area of fibrosis. We also observed that MMP-2 was transiently increased in BAL following AFSC treatment. Increased MMP-2 activity was further associated with cleavage of CCL2, rendering it a putative antagonist for CCL2/CCR2 signaling, which we surmise is a potential mechanism for CCL2 reduction in BAL following AFSC treatment. Based on this data, we concluded that AFSC have the potential to inhibit the development or progression of fibrosis in a bleomycin injury model during both acute and chronic remodeling events.

  12. Amniotic Fluid Stem Cells Inhibit the Progression of Bleomycin-Induced Pulmonary Fibrosis via CCL2 Modulation in Bronchoalveolar Lavage

    Science.gov (United States)

    Garcia, Orquidea; Carraro, Gianni; Turcatel, Gianluca; Hall, Marisa; Sedrakyan, Sargis; Roche, Tyler; Buckley, Sue; Driscoll, Barbara; Perin, Laura; Warburton, David

    2013-01-01

    The potential for amniotic fluid stem cell (AFSC) treatment to inhibit the progression of fibrotic lung injury has not been described. We have previously demonstrated that AFSC can attenuate both acute and chronic-fibrotic kidney injury through modification of the cytokine environment. Fibrotic lung injury, such as in Idiopathic Pulmonary Fibrosis (IPF), is mediated through pro-fibrotic and pro-inflammatory cytokine activity. Thus, we hypothesized that AFSC treatment might inhibit the progression of bleomycin-induced pulmonary fibrosis through cytokine modulation. In particular, we aimed to investigate the effect of AFSC treatment on the modulation of the pro-fibrotic cytokine CCL2, which is increased in human IPF patients and is correlated with poor prognoses, advanced disease states and worse fibrotic outcomes. The impacts of intravenous murine AFSC given at acute (day 0) or chronic (day 14) intervention time-points after bleomycin injury were analyzed at either day 3 or day 28 post-injury. Murine AFSC treatment at either day 0 or day 14 post-bleomycin injury significantly inhibited collagen deposition and preserved pulmonary function. CCL2 expression increased in bleomycin-injured bronchoalveolar lavage (BAL), but significantly decreased following AFSC treatment at either day 0 or at day 14. AFSC were observed to localize within fibrotic lesions in the lung, showing preferential targeting of AFSC to the area of fibrosis. We also observed that MMP-2 was transiently increased in BAL following AFSC treatment. Increased MMP-2 activity was further associated with cleavage of CCL2, rendering it a putative antagonist for CCL2/CCR2 signaling, which we surmise is a potential mechanism for CCL2 reduction in BAL following AFSC treatment. Based on this data, we concluded that AFSC have the potential to inhibit the development or progression of fibrosis in a bleomycin injury model during both acute and chronic remodeling events. PMID:23967234

  13. Amniotic fluid embolism mortality rate.

    Science.gov (United States)

    Benson, Michael D

    2017-11-01

    The objective of this study was to determine the mortality rate of amniotic fluid embolism (AFE) using population-based studies and case series. A literature search was conducted using the two key words: 'amniotic fluid embolism (AFE)' AND 'mortality rate'. Thirteen population-based studies were evaluated, as well as 36 case series including at least two patients. The mortality rate from population-based studies varied from 11% to 44%. When nine population-based studies with over 17 000 000 live births were aggregated, the maternal mortality rate was 20.4%. In contrast, the mortality rate of AFE in case series varies from 0% to 100% with numerous rates in between. The AFE mortality rate in population-based studies varied from 11% to 44% with the best available evidence supporting an overall mortality rate of 20.4%. Data from case series should no longer be used as a basis for describing the lethality of AFE. © 2017 Japan Society of Obstetrics and Gynecology.

  14. Amniotic Fluid Cells Proliferation in Normal and Down Syndrome Subjects

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    Honcea Adina

    2016-02-01

    Full Text Available Down Syndrome/Trisomy 21 is the most common chromosomal anomaly, and it represents the most common congenital cause of infants’ intellectual disability. Subjects with this syndrome are affected by degenerative processes caused by accelerated aging or unknown ethyologies. In recent years, accumulating evidence revealed increased potential of amniotic fluid-derived stem cells to be used in regenerative therapy. Our aim was to assess differences in immunophenotype, cell morphology and proliferation of amniotic fluid cells from normal and Down Syndrome pregnancies using a quantitative cytometry approach. Results revealed the emergence of a population of small sized cells in Down Syndrome derived amniotic fluid cells that are readily visible upon microscopic inspection. Hence, the fluorescence–based quantitative image cytometry determinations showed a tendency of decrease in both cell and nuclei size in trisomy, with no significant modification in nuclei circularity, as measured following actin cytoskeleton and nuclei labeling. The propensity of Ki67 positive cells was found to be increased in Down Syndrome derived cells (48.92% as compared to normal specimens (28.68%. However, cells in S and G2/M cell cycle phases decreased from 32.91% to 4.49% in diseased cells. Further studies are devoted to understanding the molecular basis of the observed differences in the proliferation ability of Down Syndrome amniotic cells, in order to evaluate the potential therapeutic effect of amniotic fluid stem cells for tissue regeneration in subjects with trisomy and to find correlations between amniotic cells phenotype and patient prognosis.

  15. Clone-derived human AF-amniotic fluid stem cells are capable of skeletal myogenic differentiation in vitro and in vivo.

    Science.gov (United States)

    Ma, Xiaorong; Zhang, Shengli; Zhou, Junmei; Chen, Baisong; Shang, Yafeng; Gao, Tongbing; Wang, Xue; Xie, Hua; Chen, Fang

    2012-08-01

    Stem cell-based therapy may be the most promising method to cure skeletal muscle degenerative diseases such as Duchenne muscular dystrophy (DMD) and trauma in the future. Human amniotic fluid is enriched with early-stage stem cells from developing fetuses and these cells have cardiomyogenic potential both in vitro and in vivo. In the present study, we investigated the characteristics of human amniotic fluid-derived AF-type stem (HAF-AFS) cells by flow cytometry, immunofluorescence staining, reverse-transcription polymerase chain reaction, and osteogenic and adipogenic differentiation analysis. After confirming the stemness of HAF-AFS cells, we tested whether HAF-AFS cells could differentiate into skeletal myogenic cells in vitro and incorporate into regenerating skeletal muscle in vivo. By temporary exposure to the DNA demethylation agent 5-aza-2'-deoxycytidine (5-Aza dC) or co-cultured with C2C12 myoblasts, HAF-AFS cells differentiated into skeletal myogenic cells, expressing skeletal myogenic cell-specific markers such as Desmin, Troponin I (Tn I) and α-Actinin. Four weeks after transplantation into cardiotoxin-injured and X-ray-irradiated tibialis anterior (TA) muscles of NOD/SCID mice, HAF-AFS cells survived, differentiated into myogenic precursor cells and fused with host myofibres. The findings that HAF-AFS cells differentiate into myogenic cells in vitro and incorporate in skeletal muscle regeneration in vivo hold the promise of HAF-AFS cell-based therapy for skeletal muscle degenerative diseases. Copyright © 2012 John Wiley & Sons, Ltd.

  16. Non-integrating episomal plasmid-based reprogramming of human amniotic fluid stem cells into induced pluripotent stem cells in chemically defined conditions.

    Science.gov (United States)

    Slamecka, Jaroslav; Salimova, Lilia; McClellan, Steven; van Kelle, Mathieu; Kehl, Debora; Laurini, Javier; Cinelli, Paolo; Owen, Laurie; Hoerstrup, Simon P; Weber, Benedikt

    2016-01-01

    Amniotic fluid stem cells (AFSC) represent an attractive potential cell source for fetal and pediatric cell-based therapies. However, upgrading them to pluripotency confers refractoriness toward senescence, higher proliferation rate and unlimited differentiation potential. AFSC were observed to rapidly and efficiently reacquire pluripotency which together with their easy recovery makes them an attractive cell source for reprogramming. The reprogramming process as well as the resulting iPSC epigenome could potentially benefit from the unspecialized nature of AFSC. iPSC derived from AFSC also have potential in disease modeling, such as Down syndrome or β-thalassemia. Previous experiments involving AFSC reprogramming have largely relied on integrative vector transgene delivery and undefined serum-containing, feeder-dependent culture. Here, we describe non-integrative oriP/EBNA-1 episomal plasmid-based reprogramming of AFSC into iPSC and culture in fully chemically defined xeno-free conditions represented by vitronectin coating and E8 medium, a system that we found uniquely suited for this purpose. The derived AF-iPSC lines uniformly expressed a set of pluripotency markers Oct3/4, Nanog, Sox2, SSEA-1, SSEA-4, TRA-1-60, TRA-1-81 in a pattern typical for human primed PSC. Additionally, the cells formed teratomas, and were deemed pluripotent by PluriTest, a global expression microarray-based in-silico pluripotency assay. However, we found that the PluriTest scores were borderline, indicating a unique pluripotent signature in the defined condition. In the light of potential future clinical translation of iPSC technology, non-integrating reprogramming and chemically defined culture are more acceptable.

  17. Amniotic fluid embolism: A diagnostic dilemma.

    Science.gov (United States)

    Kulshrestha, Ashish; Mathur, Megha

    2011-01-01

    Amniotic fluid embolism (AFE) is a rare obstetric catastrophe with an incidence of 7.7 per 100 000 deliveries and mortality as high as 60% to 80%. We describe a case of perioperative cardiac arrest in a young parturient undergoing an emergent cesarean section. Just after delivery of live healthy male baby, patient developed disseminated intravascular coagulation not responding to resuscitation with fluids and blood products. Her autopsy revealed edematous lungs with amniotic fluid debris within pulmonary vessels thus establishing the diagnosis of AFE. Amniotic fluid embolism is life threatening and difficult to predict or prevent condition, which should be always be kept in mind in a parturient with sudden cardiovascular collapse, so that resuscitation commences immediately, as early intervention is essential for a positive outcome.

  18. Glial cell line-derived neurotrophic factor induced the differentiation of amniotic fluid-derived stem cells into vascular endothelial-like cells in vitro.

    Science.gov (United States)

    Zhang, Ruyu; Lu, Ying; Li, Ju; Wang, Jia; Liu, Caixia; Gao, Fang; Sun, Dong

    2016-02-01

    Amniotic fluid-derived stem cells (AFSCs) are a novel source of stem cells that are isolated and cultured from second trimester amniocentesis. Glial cell line-derived neurotrophic factor (GDNF) acts as a tissue morphogen and regulates stem cell proliferation and differentiation. This study investigated the effect of an adenovirus-mediated GDNF gene, which was engineered into AFSCs, on the cells' biological properties and whether GDNF in combination with AFSCs can be directionally differentiated into vascular endothelial-like cells in vitro. AFSCs were isolated and cultured using the plastic adherence method in vitro and identified by the transcription factor Oct-4, which is the primary marker of pluripotent stem cells. AFSCs were efficiently transfected by a GFP-labeled plasmid system of an adenovirus vector carrying the GDNF gene (Ad-GDNF-GFP). Transfected AFSCs stably expressed GDNF. Transfected AFSCs were cultured in endothelial growth medium-2 containing vascular endothelial growth factor. After 1 week, AFSCs were positive for von Willebrand factor (vWF) and CD31, which are markers of endothelial cells, and the recombinant GDNF group was significantly higher than undifferentiated controls and the GFP only group. These results demonstrated that AFSCs differentiated into vascular endothelial-like cells in vitro, and recombinant GDNF promoted differentiation. The differentiation-induced AFSCs may be used as seed cells to provide a new manner of cell and gene therapies for transplantation into the vascular injury site to promote angiogenesis.

  19. Role of Echogenic Amniotic Fluid Particles and Optical Density in ...

    African Journals Online (AJOL)

    This study was aimed to correlate echogenic amniotic fluid particle size (AFPS) in late third trimester to fetal lung maturity and amniotic fluid optical density (AFOD) at labor. AFPS were measured with specified criteria by real time transabdominal USG (3.5MHz) while Amniotic Fluid Index (AFI) was measured during routine ...

  20. Amniotic fluid stem cells restore the muscle cell niche in a HSA-Cre, Smn(F7/F7) mouse model.

    Science.gov (United States)

    Piccoli, Martina; Franzin, Chiara; Bertin, Enrica; Urbani, Luca; Blaauw, Bert; Repele, Andrea; Taschin, Elisa; Cenedese, Angelo; Zanon, Giovanni Franco; André-Schmutz, Isabelle; Rosato, Antonio; Melki, Judith; Cavazzana-Calvo, Marina; Pozzobon, Michela; De Coppi, Paolo

    2012-08-01

    Mutations in the survival of motor neuron gene (SMN1) are responsible for spinal muscular atrophy, a fatal neuromuscular disorder. Mice carrying a homozygous deletion of Smn exon 7 directed to skeletal muscle (HSA-Cre, Smn(F7/F7) mice) present clinical features of human muscular dystrophies for which new therapeutic approaches are highly warranted. Herein we demonstrate that tail vein transplantation of mouse amniotic fluid stem (AFS) cells enhances the muscle strength and improves the survival rate of the affected animals. Second, after cardiotoxin injury of the Tibialis Anterior, only AFS-transplanted mice efficiently regenerate. Most importantly, secondary transplants of satellite cells (SCs) derived from treated mice show that AFS cells integrate into the muscle stem cell compartment and have long-term muscle regeneration capacity indistinguishable from that of wild-type-derived SC. This is the first study demonstrating the functional and stable integration of AFS cells into the skeletal muscle, highlighting their value as cell source for the treatment of muscular dystrophies. Copyright © 2012 AlphaMed Press.

  1. Escalated regeneration in sciatic nerve crush injury by the combined therapy of human amniotic fluid mesenchymal stem cells and fermented soybean extracts, Natto

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    Pan Hung-Chuan

    2009-08-01

    Full Text Available Abstract Attenuation of inflammatory cell deposits and associated cytokines prevented the apoptosis of transplanted stem cells in a sciatic nerve crush injury model. Suppression of inflammatory cytokines by fermented soybean extracts (Natto was also beneficial to nerve regeneration. In this study, the effect of Natto on transplanted human amniotic fluid mesenchymal stem cells (AFS was evaluated. Peripheral nerve injury was induced in SD rats by crushing a sciatic nerve using a vessel clamp. Animals were categorized into four groups: Group I: no treatment; Group II: fed with Natto (16 mg/day for 7 consecutive days; Group III: AFS embedded in fibrin glue; Group IV: Combination of group II and III therapy. Transplanted AFS and Schwann cell apoptosis, inflammatory cell deposits and associated cytokines, motor function, and nerve regeneration were evaluated 7 or 28 days after injury. The deterioration of neurological function was attenuated by AFS, Natto, or the combined therapy. The combined therapy caused the most significantly beneficial effects. Administration of Natto suppressed the inflammatory responses and correlated with decreased AFS and Schwann cell apoptosis. The decreased AFS apoptosis was in line with neurological improvement such as expression of early regeneration marker of neurofilament and late markers of S-100 and decreased vacuole formation. Administration of either AFS, or Natto, or combined therapy augmented the nerve regeneration. In conclusion, administration of Natto may rescue the AFS and Schwann cells from apoptosis by suppressing the macrophage deposits, associated inflammatory cytokines, and fibrin deposits.

  2. In vitro culture of mouse embryos amniotic fluid ID human

    African Journals Online (AJOL)

    1989-07-15

    Jul 15, 1989 ... Human amniotic fluid was compared with Ham's F-10 culture medium as a possible alternative for use in in vitro fertilisation. The cleavage success of mouse embryos in human amniotic fluid (experimental group) was 92% compared with 86% in. Ham's F-10 medium. It is concluded that human amniotic.

  3. Senescence-Associated Molecular and Epigenetic Alterations in Mesenchymal Stem Cell Cultures from Amniotic Fluid of Normal and Fetus-Affected Pregnancy

    Directory of Open Access Journals (Sweden)

    Jūratė Savickienė

    2016-01-01

    Full Text Available Human amniotic-fluid-derived mesenchymal stem cells (AF-MSCs are interesting for their multilineage differentiation potential and wide range of therapeutic applications due to the ease of culture expansion. However, MSCs undergo replicative senescence. So far, the molecular mechanisms that underlie fetal diseases and cell senescence are still poorly understood. Here, we analyzed senescence-associated morphologic, molecular, and epigenetic characteristics during propagation of MSCs derived from AF of normal and fetus-affected pregnancy. AF-MSCs cultures from both cell sources displayed quite similar morphology and expression of specific cell surface (CD44, CD90, and CD105 and stemness (Oct4, Nanog, Sox2, and Rex1 markers but had interindividual variability in proliferation capability and time to reach senescence. Within passages 4 and 8, senescent cultures exhibited typical morphological features, senescence-associated β-galactosidase activity, increased levels of p16, and decreased levels of miR-17 and miR-21 but showed differential expression of p21, p53, and ATM dependently on the onset of cell senescence. These differences correlated with changes in the level of chromatin modifiers (DNMT1 and HDAC1 and polycomb group proteins (EZH2, SUZ12, and BMI1 paralleling with changes in the expression of repressive histone marks (H3K9me3 and H3K27me3 and stemness markers (Oct4, Nanog, Sox2, and Rex1. Therefore epigenetic factors are important for AF-MSCs senescence process that may be related with individuality of donor or a fetus malignancy status.

  4. 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.

  5. Intravenous grafts of amniotic fluid-derived stem cells induce endogenous cell proliferation and attenuate behavioral deficits in ischemic stroke rats.

    Directory of Open Access Journals (Sweden)

    Naoki Tajiri

    Full Text Available We recently reported isolation of viable rat amniotic fluid-derived stem (AFS cells [1]. Here, we tested the therapeutic benefits of AFS cells in a rodent model of ischemic stroke. Adult male Sprague-Dawley rats received a 60-minute middle cerebral artery occlusion (MCAo. Thirty-five days later, animals exhibiting significant motor deficits received intravenous transplants of rat AFS cells or vehicle. At days 60-63 post-MCAo, significant recovery of motor and cognitive function was seen in stroke animals transplanted with AFS cells compared to vehicle-infused stroke animals. Infarct volume, as revealed by hematoxylin and eosin (H&E staining, was significantly reduced, coupled with significant increments in the cell proliferation marker, Ki67, and the neuronal marker, MAP2, in the dentate gyrus (DG [2] and the subventricular zone (SVZ of AFS cell-transplanted stroke animals compared to vehicle-infused stroke animals. A significantly higher number of double-labeled Ki67/MAP2-positive cells and a similar trend towards increased Ki67/MAP2 double-labeling were observed in the DG and SVZ of AFS cell-transplanted stroke animals, respectively, compared to vehicle-infused stroke animals. This study reports the therapeutic potential of AFS cell transplantation in stroke animals, possibly via enhancement of endogenous repair mechanisms.

  6. Human amniotic membrane as newly identified source of amniotic fluid pulmonary surfactant

    OpenAIRE

    Lemke, Angela; Castillo-S?nchez, Jos? Carlos; Prodinger, Florian; Ceranic, Asja; Hennerbichler-Lugscheider, Simone; P?rez-Gil, Jes?s; Redl, Heinz; Wolbank, Susanne

    2017-01-01

    Pulmonary surfactant (PS) reduces surface tension at the air-liquid interface in the alveolar epithelium of the lung, which is required for breathing and for the pulmonary maturity of the developing foetus. However, the origin of PS had never been thoroughly investigated, although it was assumed to be secreted from the foetal developing lung. Human amniotic membrane (hAM), particularly its epithelial cell layer, composes the amniotic sac enclosing the amniotic fluid. In this study, we therefo...

  7. In vitro culture of mouse embryos amniotic fluid ID human

    African Journals Online (AJOL)

    1989-07-15

    Jul 15, 1989 ... Because human amniotic fluid is a physiological, balanced ultrafiltrate, it has been considered as an inexpensive alternative culture medium in. IVF. A study of the development of mouse embryos in human amniotic fluid was undertaken to assess the suitability of this as an optional culture medium in human ...

  8. Amniotic fluid embolism in an HIV-positive parturient

    African Journals Online (AJOL)

    2010-02-01

    Feb 1, 2010 ... Case Studies: Amniotic fluid embolism in an HIV-positive parturient. 2010;16(4). S Afr J Anaesthesiol Analg. Abstract. We present a case of a parturient infected with human immunodeficiency virus, who developed amniotic fluid embolism during the delivery of her twins by elective Caesarean section.

  9. Phthalates and perfluorooctanesulfonic acid in human amniotic fluid

    DEFF Research Database (Denmark)

    Jensen, Morten Søndergaard; Nørgaard-Pedersen, Bent; Toft, Gunnar

    2012-01-01

    Measures of prenatal environmental exposures are important, and amniotic fluid levels may directly reflect fetal exposures during hypothesized windows of vulnerability.......Measures of prenatal environmental exposures are important, and amniotic fluid levels may directly reflect fetal exposures during hypothesized windows of vulnerability....

  10. Fetal Therapy Model of Myelomeningocele with Three-Dimensional Skin Using Amniotic Fluid Cell-Derived Induced Pluripotent Stem Cells

    Directory of Open Access Journals (Sweden)

    Kazuhiro Kajiwara

    2017-06-01

    Full Text Available Myelomeningocele (MMC is a congenital disease without genetic abnormalities. Neurological symptoms are irreversibly impaired after birth, and no effective treatment has been reported to date. Only surgical repairs have been reported so far. In this study, we performed antenatal treatment of MMC with an artificial skin using induced pluripotent stem cells (iPSCs generated from a patient with Down syndrome (AF-T21-iPSCs and twin-twin transfusion syndrome (AF-TTTS-iPSCs to a rat model. We manufactured three-dimensional skin with epidermis generated from keratinocytes derived from AF-T21-iPSCs and AF-TTTS-iPSCs and dermis of human fibroblasts and collagen type I. For generation of epidermis, we developed a protocol using Y-27632 and epidermal growth factor. The artificial skin was successfully covered over MMC defect sites during pregnancy, implying a possible antenatal surgical treatment with iPSC technology.

  11. Amniotic fluid embolism and isolated coagulopathy: atypical presentation of amniotic fluid embolism.

    LENUS (Irish Health Repository)

    Awad, I T

    2012-02-03

    A 41-year-old multigravida presented at 32 weeks of gestation with polyhydramnios and an anencephalic fetus. Abnormal bleeding as a result of disseminated intravascular coagulation complicated an emergency Caesarean section for severe abdominal pain thought to be due to uterine rupture. Massive transfusion with blood products was necessary and the abdomen packed to control bleeding. The patient was transferred to the intensive care unit where she made a slow but complete recovery. Amniotic fluid embolism with atypical presentation of isolated coagulopathy is the likely diagnosis in this case. The case serves to demonstrate that amniotic fluid embolism may present with symptoms and signs other than the classical pattern of dyspnoea, cyanosis and hypotension.

  12. Does reduction of amniotic fluid affect fetal movements?

    NARCIS (Netherlands)

    Sival, D A; Visser, G H; Prechtl, H F

    The effect of the amount of amniotic fluid on the form of fetal general movements was studied longitudinally in 19 pregnancies complicated by premature rupture of the amniotic membranes (PROM). Before birth, general movements were studied weekly by means of 1-h ultrasound observations, performed

  13. Pup retrieval and maternal attraction to canine amniotic fluids.

    Science.gov (United States)

    Dunbar, I; Ranson, E; Buehler, M

    1981-10-01

    Three purebred female beagles were observed with both their first and second litters. Dams were given three separate simultaneous-choice retrieval tests: In addition, the maternal response to amniotic fluids was observed when one pup in the litter was treated with amniotic fluids and all other pups were treated with water. There was no evidence to suggest that pups were retrieved preferentially on the basis of their sex. One female retrieved her own but not alien pups, whereas another female made no such discrimination and readily retrieved alien pups in addition to her own. Retrieval behavior developed at the time of each whelping and normally lasted until the pups were about 5 days old, although a bitch would retrieve younger (alien) pups up to 14 days post partum. Maternal bitches were strongly attracted towards amniotic fluids: they investigated pups daubed with amniotic fluids to a significantly greater extent than control pups treated with water. The attraction of maternal females towards amniotic fluids developed at the time of each whelping and persisted for up to 30 days, well beyond the time that a whelping bitch would normally be exposed to her own fetal fluids. A possible role for amniotic fluids in the development of maternal behaviour and the establishment of the maternal/puppy bond is discussed. Copyright © 1981. Published by Elsevier B.V.

  14. Testosterone determination in amniotic fluid for sec diagnosis

    International Nuclear Information System (INIS)

    Quiroa C, M.M.

    1985-11-01

    This study was carried on 50 samples of amniotic fluid obtained from pregnant patients with gestation old of 38 to 40 weeks; diagnosis of the foetus sex was made by measuring the testosterone levels by radioimmunoassay technique. It was found that 94% of the cases were correctly diagnosed. The testosterone levels found in the amniotic fluid of male and female foetus were significantly different (L<0.01) these confirm the efficacy of the method. (author)

  15. [Biologic tests for the diagnosis of amniotic fluid embolism].

    Science.gov (United States)

    Van Cortenbosch, B; Huel, C; Houfflin Debarge, V; Luton, D; Lambaudie, E; Porquet, D; Guibourdenche, J

    2007-01-01

    Amniotic fluid embolism is a rare, unpredictable and often lethal complication of pregnancy and childbirth. Because of its variable presentation, an early biologic test would help to establish the diagnosis. We investigated in maternal serum 4 components of amniotic fluid, i.e., alpha-fetoprotein (AFP), l'insuline like growth factor binding protein-1 (IGFBP-1), fetal fibronectin (fFN) and placental alpha1-microglobulin (PAMG-1). On the 6 cesareans controls involved, none of the makers increased after membranes section. PAMG-1 is unsuitable because its detection is always positive or doubtful even in the baseline. On the 7 cases suspected of amniotic fluid embolism, no detectable increase in any of those markers was noted in 3 cases, which is not in favour of this diagnosis. In the remaining cases, IGFBP-1 and fFN became detectable, confirming histological evidences of amniotic fluid embolism in 2 cases. The follow up of those markers in maternal blood confirmed the suspicion of amniotic fluid embolism at 21 wg in one case of ongoing pregnancy. These preliminary results point out the potential interest to assay maternal serum AFP, IGFBP-1 and fFN to confirm amniotic fluid embolism using rapid laboratory tests.

  16. Human amniotic membrane as newly identified source of amniotic fluid pulmonary surfactant.

    Science.gov (United States)

    Lemke, Angela; Castillo-Sánchez, José Carlos; Prodinger, Florian; Ceranic, Asja; Hennerbichler-Lugscheider, Simone; Pérez-Gil, Jesús; Redl, Heinz; Wolbank, Susanne

    2017-07-25

    Pulmonary surfactant (PS) reduces surface tension at the air-liquid interface in the alveolar epithelium of the lung, which is required for breathing and for the pulmonary maturity of the developing foetus. However, the origin of PS had never been thoroughly investigated, although it was assumed to be secreted from the foetal developing lung. Human amniotic membrane (hAM), particularly its epithelial cell layer, composes the amniotic sac enclosing the amniotic fluid. In this study, we therefore aimed to investigate a potential contribution of the cellular components of the hAM to pulmonary surfactant found in amniotic fluid. We identified that cells within the native membrane contain lamellar bodies and express all four surfactant proteins as well as ABCA3. Lipidomic profiling by nanoESI - MS/MS revealed the presence of the essential lipid species as found in PS. Also, the biophysical activity of conditioned cell culture supernatant obtained from hAM was tested with captive bubble surfactometry. hAM supernatant showed the ability to reduce surface tension, similar to human PS obtained from bronchoalveolar lavage. This means that hAM produces the essential PS-associated components and can therefore contribute as second potential source of PS in amniotic fluid aside from the foetal lung.

  17. Amniotic fluid as a vital sign for fetal wellbeing

    Science.gov (United States)

    Magann, Everett F

    2015-01-01

    Abstract Introduction: Amniotic fluid, once thought to merely provide protection and room for necessary movement and growth for the fetus, is now understood to be a highly complex and dynamic system that is studied as a data point to interpret fetal wellbeing. Methods: Assessment of amniotic fluid volume is now routine when performing a sonographic evaluation of fetal status and is an important consideration in the assessment and management of perinatal morbidity and mortality. 1 , 2 In this review, we will cover the dynamics that affect amniotic fluid volume, review methods for measurement and quantification of volume, review definitions for normative data as related to neonatal outcomes, and provide evidence based guidance on the workup and management options for oligoydramnios and polyhydramnios in singleton and twin pregnancies. Conclusions: When abnormalities of fluid exist, appropriate workup to uncover the underlying etiology should be initiated as adverse fetal outcomes are sometimes associated with these variations from normalcy. PMID:28191176

  18. Trans-amniotic stem cell therapy (TRASCET) minimizes Chiari-II malformation in experimental spina bifida.

    Science.gov (United States)

    Dionigi, Beatrice; Brazzo, Joseph A; Ahmed, Azra; Feng, Christina; Wu, Yaotang; Zurakowski, David; Fauza, Dario O

    2015-06-01

    We sought to study the impact of trans-amniotic stem cell therapy (TRASCET) in the Chiari-II malformation in experimental spina bifida. Sprague-Dawley fetuses (n=62) exposed to retinoic acid were divided into three groups at term (21-22 days gestation): untreated isolated spina bifida (n=21), isolated spina bifida treated with intra-amniotic injection of concentrated, syngeneic, labeled amniotic fluid mesenchymal stem cells (afMSCs) on gestational day 17 (n=28), and normal controls (n=13). Analyses included measurements of brainstem and cerebellar placement on high resolution MRI and histology. Statistical comparisons included ANOVA. In parallel to the expected induced coverage of the spina bifida in the afMSC-treated group (Pspina bifida by TRASCET minimizes the Chiari-II malformation in the retinoic acid rodent model, further suggesting it as a practical alternative for the prenatal management of spina bifida. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Staphylococcus lugdunensis cultured from the amniotic fluid at Caesarean Section.

    Directory of Open Access Journals (Sweden)

    Zbigniew Marchocki

    Full Text Available Staphylococcus lugdunensis is a virulent coagulase-negative staphylococcus. It behaves like and can be mistaken in culture for Staphylococcus aureus. While originally thought to be a skin commensal rarely responsible for opportunistic infection, it was rapidly established as a significant human pathogen. It has been mainly associated with native and prosthetic valve endocarditis, osteomyelitis, and skin and soft tissue cellulitis, but has also been reported as a cause of fasciitis as well as peritonitis. Staphylococcus lugdunensis has been reported as a cause of endometritis but has not been previously isolated from amniotic fluid. Here, amniotic fluid samples were collected in the course of a larger study on amniotic fluid bacteriology, with prior ethical approval and informed patient consent. Amniotic fluid was obtained at Caesarean Section by direct needle aspiration from the intact amnion. Analysis with Staphylococcal API test kits led to identification of Staphylococcus lugdunensis in two cases. The clinical significance of the finding in these reported cases is undetermined. Staphylococcus lugdunensis has been shown to be a cause of serious and potentially fatal morbidities, but this is the first report of its culture from amniotic fluid. As caesarean delivery is accepted as the single most important factor associated with post-partum infectious complications in both mother and neonate, the identification of this pathogen is a new concern.

  20. Isolation, Characterization, Cryopreservation of Human Amniotic Stem Cells and Differentiation to Osteogenic and Adipogenic Cells.

    Directory of Open Access Journals (Sweden)

    Shiva Gholizadeh-Ghaleh Aziz

    Full Text Available Human stem cells and progenitor cells can be used to treat cancer and replace dysfunctional cells within a tissue or organ. The objective of this study was to identify the appropriate cells type in regenerative medicine and targeted therapy. As an alternative to embryonic and bone marrow stem cells, we examined human amniotic fluid stem cells (hAFSCs, one of the potential source of multipotent stem cells isolated from both cell pellet (using single-stage method, and supernatant of human amniotic fluid. Source of isolation and unique property of the cells emphasize that these cells are one of the promising new tools in therapeutic field. Double sources for isolation and availability of the left over samples in diagnostic laboratory at the same time have less legal and ethical concerns compared with embryonic stem cell studies. Cells were isolated, cultured for 18th passage for 6 months and characterized using qPCR and flow cytometry. Cells showed good proliferative ability in culture condition. The cells successfully differentiated into the adipogenic and osteogenic lineages. Based on these findings, amniotic fluid can be considered as an appropriate and convenient source of human amniotic fluid stem cells. These cells provide potential tools for therapeutic applications in the field of regenerative medicine. To get a better understanding of crosstalk between Oct4/NANOG with osteogenesis and adipogenesis, we used network analysis based on Common Targets algorithm and Common Regulators algorithm as well as subnetwork discovery based on gene set enrichment. Network analysis highlighted the possible role of MIR 302A and MIR let-7g. We demonstrated the high expression of MIR 302A and low expression of MIR let7g in hAFSCs by qPCR.

  1. Dielectric properties of human placenta, umbilical cord and amniotic fluid

    Science.gov (United States)

    Peyman, A.; Gabriel, C.; Benedickter, H. R.; Fröhlich, J.

    2011-04-01

    The dielectric properties of freshly delivered human placenta, umbilical cord and amniotic fluid have been acquired at 37 °C and in the frequency range of 200 MHz-10 GHz. The experimental data were fitted to a Cole-Cole expression. The results show that dielectric properties of the umbilical cord are significantly higher than placenta due to the presence of high water content Wharton's jelly. The results also demonstrate large differences in the dielectric properties of amniotic and cerebrospinal fluids. The data presented can be used in numerical simulations of the exposure of pregnant women to electromagnetic fields.

  2. Dielectric properties of human placenta, umbilical cord and amniotic fluid

    Energy Technology Data Exchange (ETDEWEB)

    Peyman, A [Physical Dosimetry Department, Health Protection Agency, Chilton, Didcot OX11 0RQ (United Kingdom); Gabriel, C [MCL-P, Newbury RG14 5PY, Berkshire (United Kingdom); Benedickter, H R; Froehlich, J, E-mail: Azadeh.peyman@hpa.org.uk [Electromagnetic Fields and Microwave Electronics Laboratory, Swiss Federal Institute of Technology, Zurich (Switzerland)

    2011-04-07

    The dielectric properties of freshly delivered human placenta, umbilical cord and amniotic fluid have been acquired at 37 deg. C and in the frequency range of 200 MHz-10 GHz. The experimental data were fitted to a Cole-Cole expression. The results show that dielectric properties of the umbilical cord are significantly higher than placenta due to the presence of high water content Wharton's jelly. The results also demonstrate large differences in the dielectric properties of amniotic and cerebrospinal fluids. The data presented can be used in numerical simulations of the exposure of pregnant women to electromagnetic fields. (note)

  3. In vitro culture of mouse embryos in human amniotic fluid | Coetzee ...

    African Journals Online (AJOL)

    Human amniotic fluid was compared with Ham's F-10 culture medium as a possible alternative for use in in vitro fertilisation. The cleavage success of mouse embryos in human amniotic fluid (experimental group) was 92% compared with 86% in Ham's F-10 medium. It is concluded that human amniotic fluid is a viable ...

  4. Amniotic Fluid MMP-9 and Neurotrophins in Autism Spectrum Disorders

    DEFF Research Database (Denmark)

    Abdallah, Morsi; Pearce, Brad D; Larsen, Nanna

    2012-01-01

    Evidence suggests that some developmental disorders, such as autism spectrum disorders (ASDs), are caused by errors in brain plasticity. Given the important role of matrix metalloproteinases (MMPs) and neurotrophins (NTs) in neuroplasticity, amniotic fluid samples for 331 ASD cases and 698...

  5. Effects of fructose feeding on maternal and amniotic fluid ...

    African Journals Online (AJOL)

    Dr Olaleye

    pregnancies using corticotrophin releasing hormone. (CRH), a stress marker produced by the placenta and C. - reactive protein (CRP) an inflammatory response marker by evaluating the effect of fructose-induced insulin resistance on maternal serum and amniotic fluid insulin, CRH, CRP, IGF-1 and IGF-2 on days 12, 15.

  6. Effects of fructose feeding on maternal and amniotic fluid ...

    African Journals Online (AJOL)

    Effects of fructose feeding on maternal and amniotic fluid corticotropin releasing hormone and C-reactive protein in pregnant female Sprague-dawley rats. ... The results suggest that stress and inflammatory mechanisms may be implicated in the role maternal nutrition plays in developmental plasticity. Keywords: Insulin ...

  7. Clinical relevance of sonographically estimated amniotic fluid volume: polyhydramnios.

    Science.gov (United States)

    Sandlin, Adam T; Chauhan, Suneet P; Magann, Everett F

    2013-05-01

    Polyhydramnios is an excessive amount of amniotic fluid within the amniotic cavity. The etiology of polyhydramnios may be idiopathic, the consequence of fetal structural anomalies, or the consequence of various fetal and maternal conditions. The clinical importance of polyhydramnios is found in its association with adverse pregnancy outcomes and the risk of perinatal mortality. The antenatal management of polyhydramnios can be challenging as there are no formalized guidelines on the topic. The purpose of this review is to provide a literature-based overview on the subject of polyhydramnios in singleton pregnancies, demonstrate its clinical implications, and describe a practical approach to its management.

  8. Clinical significance of amniotic fluid glucose concentration in polyhydramnios.

    Science.gov (United States)

    Ogawa, Masaki; Sato, Akira; Mori, Koutaro; Nagao, Daisuke; Kamada, Kumiko; Sato, Megumi; Terada, Yukihiro

    2012-07-01

    To evaluate whether amniotic fluid glucose concentration can predict poor neonatal outcome in cases with polyhydramnios as an overall assessment. In this retrospective study, we have reviewed 42 consecutive pregnant women with singleton gestations complicated with polyhydramnios who gave birth at one tertiary care center between January 2003 and September 2010. Perinatal clinical findings were reviewed, and the neonatal outcome, such as livebirth or stillbirth and early neonatal death, was compared. A p value less than 0.05 was considered to be significant. Thirteen of 42 neonates had a poor outcome, including stillbirth or early neonatal death. Multiple logistic regression analysis showed that amniotic fluid glucose (p = 0.042) was significant factor influencing poor neonatal prognosis [odds ratio 0.66; 95% confidence interval 0.44-0.98]. Receiver operating characteristics curve and sensitivity and specificity curve showed that the optimal cut-off value of amniotic fluid glucose concentration for predicting poor neonatal outcome was 17 mg/dl. Amniotic glucose concentration less than 17 mg/dl and the presence of congenital heart anomaly were better predictors for a poor postnatal outcome in cases with polyhydramnios.

  9. Carcinoembryonic antigen as a biomarker for meconium-stained amniotic fluid.

    Science.gov (United States)

    Mor, Amir; Tal, Reshef; Irani, Mohamad; McCalla, Sandra; Haberman, Shoshana; Garg, Deepika; Wajntraub, Birgitta

    2016-03-01

    To assess whether elevated carcinoembryonic antigen (CEA) concentration in amniotic fluid can indicate meconium-stained amniotic fluid (MSAF). In a prospective cohort study, women with a term singleton pregnancy who were in labor but had intact membranes were recruited at a center in Israel over a 5-month period in 2013. Only women who subsequently underwent artificial rupture of membranes following a clear medical indication were included. Samples of amniotic fluid, urine, and serum were collected. Amniotic fluid was examined by sight and classified as clear, MSAF, or undetermined. CEA concentration in the samples was measured. Among 81 participants, 45 had clear amniotic fluid, 28 had MSAF, and eight had undetermined amniotic fluid. Mean CEA concentration was more than 10 times higher in MSAF (2658 μg/L, standard error 250) than in clear amniotic fluid (238 μg/L, standard error 29; P<0.001). Receiver operating characteristic curve analysis demonstrated a sensitivity of 96% and a specificity of 100% for distinguishing MSAF from clear amniotic fluid at a CEA cutoff of 799.2 μg/L. CEA concentrations in urine and serum were all within the normal range (≤5 μg/L), irrespective of amniotic fluid status. High CEA concentrations in amniotic fluid can assist in the diagnosis of MSAF. These findings could provide the basis for a bedside test to detect MSAF following rupture of membranes. Copyright © 2015 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

  10. RIA of alpha-fetoprotein in serum and amniotic fluid

    International Nuclear Information System (INIS)

    Fingerova, H.; Talas, M.; Stroufova, A.

    1979-01-01

    An own modification of the double antibody radioimmunoassay for AFP using 125 I-labelled AFP as a tracer, rabbit anti-AFP obtained from SEVAC, Prague and precipitating antibodies prepared by the authors is described. The AFP levels measured in the serum and the amniotic fluid using the method were in agreement with those obtained by the means of the AFPK RIA kit by SORIN in the Institute for the Care of Mother and Child in Prague. The AFP concentrations found in the cord serum and the amniotic fluid were confirmed also by the rocket electroimmunoassay according to Laurell. The described AFP RIA seems suitable for the clinical application in prenatal screening for congenital malformations, in difficult pregnancies, in hepatology and the diagnosis and the evaluation of therapy of some human malignancies. (author)

  11. Amniotic Fluid Glucose Concentration: A Marker for Infection in Preterm Labor and Preterm Premature Rupture of Membranes

    Directory of Open Access Journals (Sweden)

    Gary A. Dildy

    1994-01-01

    Full Text Available Amniotic fluid Gram stain and culture have been utilized as laboratory tests of microbial invasion of the amniotic cavity. The Gram stain of amniotic fluid has a low sensitivity in the detection of clinical infection or microbial invasion of the amniotic cavity, and amniotic fluid culture results are not immediately available for management decisions. Glucose concentration is used to diagnose infection in other sites such as cerebrospinal fluid.

  12. Results of six years of cytogenetic studies in amniotic fluid

    Directory of Open Access Journals (Sweden)

    Enelis Reyes Reyes

    2015-10-01

    Full Text Available Background: research into different genetic diseases is one of the preventive programs of paramount importance at public health level. The early detection of chromosomopathies and the establishment of an appropriate strategy reduce the morbidity-morality rate and improve the patients’ quality of life.Objective: to describe the behavior of the results of the cytogenetic studies in the amniotic fluid of pregnant women from Las Tunas province during six years: from 2008 to 2014.Methods: a retrospective and descriptive study was carried out to assess the results of cytogenetic studies in amniotic liquid during six years: from 2008 to 2014. The statistical records were checked and the results, the indication criteria, the behavior of the age groups in women advanced in age and the diagnosed chromosomopathies were assessed.Results: the samples with results that exceeded the non-conclusive and positive women prevailed; 2, 3 positive cases of chromosomopathies were diagnosed out of 100 studied women at risk; pregnant women of advanced gestational years prevailed as indication criterion, being the 37 to 40 years old age group the predominant one; in the positive cases, numeric chromosomopathies of the type trisomy 21 or Down’s syndrome prevailed, with a frequency of 1, 2 out of 100 pregnant women at risk.Conclusions: the program of the cytogenetic diagnosis in the amniotic fluid has been an effective tool to detect congenital prenatal defects by chromosomopathies, very useful in the process of genetic advice.

  13. Amniotic fluid cathepsin-G in pregnancies complicated by the preterm prelabor rupture of membranes.

    Science.gov (United States)

    Musilova, Ivana; Andrys, Ctirad; Drahosova, Marcela; Soucek, Ondrej; Pliskova, Lenka; Stepan, Martin; Bestvina, Tomas; Maly, Jan; Jacobsson, Bo; Kacerovsky, Marian

    2017-09-01

    The aim of this study was to evaluate the amniotic fluid cathepsin-G concentrations in women with preterm prelabor rupture of membranes (PPROM) based on the presence of the microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI). A total of 154 women with singleton pregnancies complicated by PPROM were included in this study. Amniotic fluid samples were obtained by transabdominal amniocentesis. Amniotic fluid cathepsin-G concentrations were assessed by ELISA. MIAC was determined using a non-cultivation approach. IAI was defined as an amniotic fluid bedside interleukin-6 concentration ≥ 745 pg/mL. Women with MIAC had higher amniotic fluid cathepsin-G concentrations than women without MIAC (with MIAC: median 82.7 ng/mL, versus without MIAC: median 64.7 ng/mL; p = 0.0003). Women with IAI had higher amniotic fluid cathepsin-G concentrations than women without this complication (with IAI: median 103.0 ng/mL, versus without IAI: median 66.2 ng/mL; p G concentrations than women with colonization (MIAC without IAI) and women without both MIAC and IAI (p G concentrations in pregnancies complicated by PPROM. Amniotic fluid cathepsin-G appears to be a potential marker of IAI.

  14. Term amniotic fluid: an unexploited reserve of mesenchymal stromal cells for reprogramming and potential cell therapy applications.

    Science.gov (United States)

    Moraghebi, Roksana; Kirkeby, Agnete; Chaves, Patricia; Rönn, Roger E; Sitnicka, Ewa; Parmar, Malin; Larsson, Marcus; Herbst, Andreas; Woods, Niels-Bjarne

    2017-08-25

    Mesenchymal stromal cells (MSCs) are currently being evaluated in numerous pre-clinical and clinical cell-based therapy studies. Furthermore, there is an increasing interest in exploring alternative uses of these cells in disease modelling, pharmaceutical screening, and regenerative medicine by applying reprogramming technologies. However, the limited availability of MSCs from various sources restricts their use. Term amniotic fluid has been proposed as an alternative source of MSCs. Previously, only low volumes of term fluid and its cellular constituents have been collected, and current knowledge of the MSCs derived from this fluid is limited. In this study, we collected amniotic fluid at term using a novel collection system and evaluated amniotic fluid MSC content and their characteristics, including their feasibility to undergo cellular reprogramming. Amniotic fluid was collected at term caesarean section deliveries using a closed catheter-based system. Following fluid processing, amniotic fluid was assessed for cellularity, MSC frequency, in-vitro proliferation, surface phenotype, differentiation, and gene expression characteristics. Cells were also reprogrammed to the pluripotent stem cell state and differentiated towards neural and haematopoietic lineages. The average volume of term amniotic fluid collected was approximately 0.4 litres per donor, containing an average of 7 million viable mononuclear cells per litre, and a CFU-F content of 15 per 100,000 MNCs. Expanded CFU-F cultures showed similar surface phenotype, differentiation potential, and gene expression characteristics to MSCs isolated from traditional sources, and showed extensive expansion potential and rapid doubling times. Given the high proliferation rates of these neonatal source cells, we assessed them in a reprogramming application, where the derived induced pluripotent stem cells showed multigerm layer lineage differentiation potential. The potentially large donor base from caesarean section

  15. Peptidome analysis of amniotic fluid from pregnancies with preeclampsia.

    Science.gov (United States)

    Qian, Yating; Zhang, Lei; Rui, Can; Ding, Hongjuan; Mao, Pengyuan; Ruan, Hongjie; Jia, Ruizhe

    2017-11-01

    Preeclampsia (PE), a life‑threatening, complicated pregnancy‑associated disease, has recently become a research focus in obstetrics. However, the peptidome of the amniotic fluid in PE patients has rarely been investigated. The present study used peptidomic profiling to perform a comparative analysis of human amniotic fluid between normal and PE pregnancies. Centrifugal ultrafiltration and liquid chromatography‑tandem mass spectrometry (LC‑MS/MS) was combined with isotopomeric dimethyl labels to gain a deeper understanding of the role of proteins and the peptidome in the onset of PE. Following ultrafiltration and LC‑MS/MS, 352 peptides were identified. Of these, 23 peptides were observed to be significantly differentially expressed (6 downregulated and 17 upregulated; Pamniotic fluid of PE and normal pregnancies. A comprehensive peptidome analysis is more efficient than a simple biomarker analysis at revealing deficiencies and improving the detection rate in diseases. These analyses therefore provide a substantial advantage in applications aimed at the discovery of disease‑specific biomarkers.

  16. Human β-Defensin-2: A natural anti-microbial peptide present in amniotic fluid participates in the host response to microbial invasion of the amniotic cavity

    Science.gov (United States)

    Soto, Eleazar; Espinoza, Jimmy; Nien, Jyh Kae; Kusanovic, Juan; Erez, Offer; Richani, Karina; Santolaya-Forgas, Joaquin; Romero, Roberto

    2012-01-01

    OBJECTIVE Human β-defensin-2 (HBD-2) is a potent anti-microbial peptide that is part of the innate immune response. The purpose of this study was to determine whether HBD-2 is present in amniotic fluid and if its concentration changes with microbial invasion of the amniotic cavity (MIAC) and labor. STUDY DESIGN Amniotic fluid was retrieved by amniocentesis from 318 patients in the following groups: 1) mid-trimester (n=75); 2) term not in labor (n=28) and in labor (n=51); 3) preterm labor and intact membranes without MIAC who delivered at term (n=36), who delivered preterm without MIAC (n=52), and preterm labor with MIAC who delivered preterm (n=25); and 4) preterm premature rupture of membranes (PPROM) with (n=25) and without MIAC (n=26). MIAC was defined as a positive amniotic fluid culture for microorganisms. Amniotic fluid HBD-2 concentrations were determined using a sensitive and specific ELISA. Non-parametric statistics were used for analysis. RESULTS 1) HBD-2 was detected in all amniotic fluid samples; 2) the concentration of HBD-2 did not change with gestational age from midtrimester to term (p=0.8); 3) Intra-amniotic infection was associated with a significant increase in amniotic fluid concentrations of HBD-2 in both women with preterm labor and intact membranes, and women with preterm PROM (p100 cell per ml) had a higher median amniotic fluid concentration of HBD-2 than those without this condition (panti-microbial peptide, and this may account for some of the anti-microbial activity of amniotic fluid; 2) Amniotic fluid HBD-2 concentrations are increased in women with MIAC, regardless of the membrane status (intact membranes or PROM); and 3) We propose that amniotic fluid HBD-2 is part of the innate immune system within the amniotic cavity. PMID:17437194

  17. Does amniotic fluid volume affect fetofetal transfusion in monochorionic twin pregnancies? Modelling two possible mechanisms

    NARCIS (Netherlands)

    Umur, Asli; van Gemert, Martin J. C.; Ross, Michael G.

    2002-01-01

    Clinical evidence suggests that increased amniotic fluid volume due to polyhydrarnnios increases placental vascular resistance. We have sought to model the possible effects of an increased amniotic fluid volume oil the net fetofetal transfusion in monochorionic twin pregnancies. We wanted to compare

  18. A cryopreservative procedure for storing cultivated and uncultivated amniotic fluid cells in liquid nitrogen.

    OpenAIRE

    Pentz, S; Hörler, H

    1980-01-01

    The cryopreservation of cultured and uncultured amniotic fluid cells is of special importance in prenatal diagnosis. The freezing procedure presented gave a cell recovery of about 95% for cultivated human amniotic fluid cells combined with a rapid appearance of mitosis after inoculation.

  19. A study of chromosomal aberrations in amniotic fluid cell cultures.

    Science.gov (United States)

    Wolstenholme, J; Crocker, M; Jonasson, J

    1988-06-01

    This paper represents the analysis of 1916 routine amniotic fluid specimens harvested by an in situ fixation technique in a prospective study with regard to cultural chromosome anomalies. Excluding constitutional abnormalities, 2.9 per cent of 19,432 cells analysed showed some form of chromosome anomaly, terminal deletions (57 per cent) and chromatid/chromosome breaks and gaps (18 per cent) being the most frequent, followed by interchange aberrations (13 per cent) and trisomy (5 per cent). No case was found of more than one colony from the same culture showing the same anomaly without it being present in other cultures from the same fluid. The wholly abnormal colonies had a surplus of trisomies and from the mathematical considerations presented one may infer that these are likely to reflect the presence of abnormal cells in the amniotic fluid. Partly abnormal colonies appeared at a frequency that would correspond to virtual absence of selection against chromosomally abnormal cells when cultured in vitro. The aberrations found were similar to those seen as single cell anomalies, except for chromatid breaks and exchanges. The data suggest a basic preferential induction of trisomy for chromosomes 2, 18, 21, and the Y-chromosome. Structural aberrations showed a marked clustering of breakpoints around the centromeres. The frequency of mutant cells was low (1.4 X 10(-3)) before culture was initiated. At harvest, the frequency of abnormal cells was much higher (3 X 10(-2)) corresponding to 3 X 10(-3) mutations per cell per generation accumulating over approximately ten generations in vitro.

  20. Antibiotics for neonates born through meconium-stained amniotic fluid.

    Science.gov (United States)

    Kelly, Lauren E; Shivananda, Sandesh; Murthy, Prashanth; Srinivasjois, Ravisha; Shah, Prakeshkumar S

    2017-06-28

    Approximately 1 in 10 pregnancies is affected by meconium passage at delivery, which can result in meconium aspiration syndrome (MAS). MAS can cause respiratory complications and, very rarely, death. Antibiotics have been prescribed for neonates exposed to meconium in amniotic fluid, with the intention of preventing infection due to potential bacterial contaminants. We conducted this review to assess the efficacy and safety of antibiotics for:1. prevention of infection, morbidity, and mortality among infants born through meconium-stained amniotic fluid (MSAF) who are asymptomatic at birth; and2. prevention of infection, morbidity, and mortality among infants born through MSAF who have signs and symptoms compatible with meconium aspiration syndrome (MAS). We performed a literature search using the following databases: MEDLINE (1966 to July 2016); Embase (1980 to July 2016); the Cumulative Index to Nursing and Allied Health Literature (CINAHL; 1982 to July 2016); and the Cochrane Central Register of Controlled Trials (CENTRAL; 2016, Issue 7) in the Cochrane Library. We also searched clinical trials databases, conference proceedings, and reference lists of retrieved articles. We included randomised and quasi-randomised controlled trials that compared antibiotics administered via any route versus placebo or no treatment for prevention of infection among neonates exposed to MSAF, or who developed MAS. We excluded cohort, case control, and any other non-randomised studies and applied no language restrictions. We included studies of term and preterm infants, and we included studies examining use of any antibacterial antibiotics. We included studies that reported on any outcomes of interest. We assessed the methodological quality of included trials by reviewing information provided in study reports and obtained by personal communication with study authors. We extracted data on relevant outcomes, estimated effect size, and reported values as risk ratios (RRs), risk

  1. Amniotic fluid and development of the immature intestine

    DEFF Research Database (Denmark)

    Østergaard, Mette Viberg

    in demand. Amniotic Fluid (AF) is the natural source of fetal EN throughout gestation in ammals. Fetal AF swallowing stimulates somatic and gastrointestinal growth during fetal development, and modulates the development of the intestinal mucosa. In addition, AF protects the fetus against infections......Premature birth is a health concern worldwide as it is associated with increased morbidity and mortality of the newborn. In the most developed countries, improved neonatal care has allowed for the survival of ever smaller premature infants and raised the survival rates of infants born very...... and maintains homeostasis in the amnion sac through anti-inflammatory mechanisms. The growth promoting and protective effects of AF in utero are ascribed to bioactive proteins including growth factors, anti-inflammatory cytokines, and antimicrobial peptides, which may exert similar beneficial effects ex utero...

  2. Amniotic fluid phthalate levels and male fetal gonad function

    DEFF Research Database (Denmark)

    Jensen, Morten Søndergaard; Anand-Ivell, Ravinder; Nørgaard-Pedersen, Bent

    2015-01-01

    BACKGROUND: Prenatal exposure to phthalates may pose a threat to human male reproduction. However, additional knowledge about the in vivo effect in humans is needed, and reported associations with genital abnormalities are inconclusive. We aimed to study prenatal di(2-ethylhexyl) phthalate (DEHP......) and diisononyl phthalate (DiNP) exposure in relation to cryptorchidism, hypospadias, and human fetal Leydig cell function. METHODS: We studied 270 cryptorchidism cases, 75 hypospadias cases, and 300 controls. Second-trimester amniotic fluid samples were available from a Danish pregnancy-screening biobank (n = 25......,105) covering 1980-1996. We assayed metabolites of DEHP and DiNP (n = 645) and steroid hormones (n = 545) by mass spectrometry. We assayed insulin-like factor 3 by immunoassay (n = 475) and analyzed data using linear or logistic regression. RESULTS: Mono(2-ethyl-5-carboxypentyl) phthalate (5cx-MEPP, DEHP...

  3. Amniotic fluid and development of the immature intestine

    DEFF Research Database (Denmark)

    Østergaard, Mette Viberg

    in demand. Amniotic Fluid (AF) is the natural source of fetal EN throughout gestation in ammals. Fetal AF swallowing stimulates somatic and gastrointestinal growth during fetal development, and modulates the development of the intestinal mucosa. In addition, AF protects the fetus against infections...... gastrointestinal structure and function in preterm pigs”. Accordingly, the aim was to test the effects of enteral administration of AF as MEN during PN, as a supplement to a suboptimal enteral diet, or both in a preterm pig model of NEC. To evaluate the effects of AF, NEC sensitivity, intestinal digestive...... function and innate immunity of the small intestine, and to establish an experimental model to investigate the inflammatory responses of the small intestine to microbial-associated molecular patterns (MAMPs) ex vivo. The expression of certain host defense genes was affected by gestational age...

  4. Prenatal sex determination by radioimmunoassay of testosterone with and without chromatography of the amniotic fluid

    International Nuclear Information System (INIS)

    Distler, W.; Boniver-Ollmann, U.; Tigges, J.; Terinde, R.; Claussen, U.

    1979-01-01

    Amniotic fluid testosterone measured by radioimmunoassay (RIA) without chromatography (immunoreactive testosterone) seems not to be a definitive test for prenatal sex determination in all cases. In this study testosterone (T) levels measured by RIA with chromatography of the amniotic fluid samples were compared with immunoreactive testosterone (iT) values, to determine the predictive accuracy of the two methods. In 111 amniotic fluid samples between 15 and 19 weeks of gestation iT and T were measured parallelly. There are significant differences between iT- and T-means of both sexes (p [de

  5. A comparison between placental and amniotic mesenchymal stem cells for transamniotic stem cell therapy (TRASCET) in experimental spina bifida.

    Science.gov (United States)

    Feng, Christina; D Graham, Christopher; Connors, John Patrick; Brazzo, Joseph; Zurakowski, David; Fauza, Dario O

    2016-06-01

    We compared placental-derived and amniotic fluid-derived mesenchymal stem cells (pMSCs and afMSCs, respectively) in transamniotic stem cell therapy (TRASCET) for experimental spina bifida. Pregnant dams (n=29) exposed to retinoic acid for the induction of fetal spina bifida were divided into four groups. Three groups received volume-matched intraamniotic injections of either saline (n=38 fetuses) or a suspension of 2×10(6) cells/mL of syngeneic, labeled afMSCs (n=73) or pMSCs (n=115) on gestational day 17 (term=21-22days). Untreated fetuses served as controls. Animals were killed before term. Statistical comparisons were by Fisher's exact test (pcell source for TRASCET as a potential alternative in the prenatal management of spina bifida. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Matrix metalloproteinase-2 is elevated in midtrimester amniotic fluid prior to the development of preeclampsia

    Directory of Open Access Journals (Sweden)

    Daniel-Spiegel Etty

    2009-08-01

    Full Text Available Abstract Objective To evaluate levels of matrix metalloproteinases (MMP and their inhibitors (TIMP in second trimester amniotic fluid of women with hypertensive disorders compared to normotensive women. Study Design Amniotic fluid was obtained from 133 women undergoing genetic second trimester amniocentesis. Zymography was performed for MMP characterization and an MMP-2 ELISA kit was used to determine MMP-2 levels. TIMP-2 expression was evaluated using western blot. Results Mean amniotic fluid MMP-2 and TIMP-2 levels were significantly higher in women who developed a hypertensive disorder compared to normotensive women (P Conclusion Higher amniotic fluid MMP-2 and TIMP-2 levels are found in women who eventually develop preeclampsia.

  7. Consequences of meconium stained amniotic fluid: What does the evidence tell us?

    NARCIS (Netherlands)

    Hutton, E.K.; Thorpe, J.

    2014-01-01

    Background: Meconium stained amniotic fluid (MSAF) is common and associated with meconium aspiration syndrome (MAS). Other consequences of meconium passage before birth are less well understood. Methods: We reviewed the literature for original papers reporting on outcomes associated with MSAF.

  8. Maternal and fetal characteristics associated with meconium-stained amniotic fluid

    DEFF Research Database (Denmark)

    Balchin, Imelda; Whittaker, John C; Lamont, Ronald F

    2011-01-01

    To estimate the rates of meconium-stained amniotic fluid (AF) and adverse outcome in relation to gestational age and racial group, and to investigate the predictors of meconium-stained AF.......To estimate the rates of meconium-stained amniotic fluid (AF) and adverse outcome in relation to gestational age and racial group, and to investigate the predictors of meconium-stained AF....

  9. Peptidomic Analysis of Amniotic Fluid for Identification of Putative Bioactive Peptides in Ventricular Septal Defect

    OpenAIRE

    Xing Li; Li-Jie Wu; Meng Gu; Yu-Mei Chen; Qi-Jun Zhang; Hua Li; Zi-Jie Cheng; Ping Hu; Shu-Ping Han; Zhang-Bin Yu; Ling-Mei Qian

    2016-01-01

    Background: Ventricular septal defect (VSD) is one of the most common congenital heart diseases and to date the role of peptides in human amniotic fluid in the pathogenesis of VSD have been rarely investigated. Methods: To gain insight into the mechanisms of protein and peptides in cardiovascular development, we constructed a comparative peptidomic profiling of human amniotic fluid between normal and VSD fetuses using a stable isobaric labeling strategy involving tandem mass tag reagents, fol...

  10. Amniotic fluid paraoxonase-1 activity, thyroid hormone concentration and oxidant status in neural tube defects.

    Science.gov (United States)

    Sak, Sibel; Agacayak, Elif; Tunc, Senem Yaman; Icen, Mehmet Sait; Findik, Fatih Mehmet; Sak, Muhammet Erdal; Yalinkaya, Ahmet; Gul, Talip

    2016-09-01

    The aim of this study was to investigate the potential association between neural tube defects and paraoxonase-1 activity in amniotic fluid. We studied total oxidant status, total antioxidant capacity, paraoxonase-1 activity and thyroid hormone amniotic fluid concentration in fetuses with neural tube defects. The present study was performed at the Department of Obstetrics and Gynaecology and the Department of Clinical Biochemistry of Dicle University between September 2011 and June 2013. The study group included 37 amniotic fluid samples from pregnant women (16-20 weeks of gestation) with fetuses affected by neural tube defects. The control group consisted of 36 pregnant women who were diagnosed with a high-risk pregnancy according to first or second trimester aneuploidy screening and were later confirmed on amniocentesis to have genetically normal fetuses. Amniotic fluid paraoxonase-1 activity and total oxidant status were significantly higher (P = 0.023, P = 0.029, respectively) whereas free T4 was significantly lower (P = 0.022) in fetuses with neural tube defects compared with control subjects. In fetuses with neural tube defects, amniotic fluid paraoxonase-1 activity correlated positively with total oxidant status (r = 0.424**, P = 0.010), and amniotic fluid total antioxidant capacity correlated positively with free t4 (r = 0.381*, P = 0.022). This is the first study in the literature to show an association between paraoxonase-1 activity and thyroid hormone concentration and neural tube defects. © 2016 Japan Society of Obstetrics and Gynecology.

  11. An assessment of the accuracy of visual diagnosis of meconium-stained amniotic fluid

    International Nuclear Information System (INIS)

    Sanlialp, C.; Caglar, G.S.; Tapisiz, O.L.; Avsar, A.F.

    2004-01-01

    Objective: The assessment of meconium content in the amniotic fluid depends on visual observation by clinicians at the bedside. The aim of the present study was to compare visual evaluation of meconium-stained amniotic fluid with spectrophotometer evaluation. Study Design: Ten gram of meconium was added to 100 ml of amniotic fluid and mixed. The solution was serially two-fold diluted with amniotic fluid. The serially diluted tubes' absorbance spectrum was measured at 420 nm and thus a standard scale was established. Ninety five samples of meconium- stained amniotic fluid were collected from labouring women and the grade of meconium was deter- mined visually at the bedside. The samples' absorbance spectrum was measured at 420 nm and recorded. Spectrophotometer was considered gold standard and the ranges of optical density in the standard scale was used to test the accuracy of visual categorization of the samples. In the statistical analysis chi-square test was used and significance was p<0.05. Results: The accuracy rate of visual diagnosis of meconium-stained amniotic fluid were found as statistically significant (accuracy rate=54.74%, p<0.001). Visual evaluation was correct in 19.4% of thin, 53.1 % of moderate and 90.6% of thick meconium samples when examined with spectrophotometer. Conclusion: Visually diagnosed thin meconium can be moderate or thick meconium when examined objectively. The visual diagnosis at bedside is not always reliable and should be replaced with an objective method like spectrophotometry. (author)

  12. Nicotine and its metabolites in amniotic fluid at birth--assessment of prenatal tobacco smoke exposure.

    Science.gov (United States)

    Köhler, E; Avenarius, S; Rabsilber, A; Gerloff, C; Jorch, G

    2010-05-01

    Amniotic fluid was collected from 78 pregnant women at birth additionally with their urine prior to delivery as well as neonatal urine and meconium. The smoking markers, nicotine and its metabolites cotinine and trans-3'-hydroxycotinine (OH-cotinine), were determined using high-performance liquid chromatography (HPLC). The self-reported smoking status during pregnancy determined by means of a questionnaire was verified by measurement of maternal urine. In all smokers, nicotine metabolites were detected in amniotic fluid and in 80% of them nicotine as well. However, the sum of the nicotine metabolites (Sum(met)) was significantly lower (p amniotic fluid (704 +/- 464 nmol/L) than in meconium (921 +/- 588 nmol/L), neonatal urine (1139 +/- 813 nmol/L) and maternal urine (4496 +/- 3535 nmol/L). Concentrations of nicotine metabolites in amniotic fluid correlated well (p amniotic fluid but only in maternal and neonatal urine. Analysis of amniotic fluid at birth lends itself to verifying smoking habits during pregnancy and clearly discriminating from ETS exposure, but it is not a suitable approach to differentiating between ETS exposure and non-exposure.

  13. Amniotic fluid myeloperoxidase in pregnancies complicated by preterm prelabor rupture of membranes.

    Science.gov (United States)

    Kacerovsky, Marian; Tambor, Vojtech; Vajrychová, Marie; Lenco, Juraj; Hornychova, Helena; Musilova, Ivana; Menon, Ramkumar

    2013-03-01

    To determine amniotic fluid myeloperoxidase concentration in women with preterm prelabor rupture of the membranes with microbial invasion of the amniotic cavity and histological chorioamnionitis. One hundred eighty-one women with singleton pregnancies with a gestational age between 24+0 and 36+6 weeks were included in this study. Amniocenteses were performed, and myeloperoxidase concentration in the amniotic fluid was determined using ELISA. Women with microbial invasion of the amniotic cavity had higher median myeloperoxidase concentration than women without this condition (149.2 ng/mL vs. 54.6 ng/mL; p = 0.0006). Women with the presence of histological chorioamnionitis had higher median myeloperoxidase concentration than women without histological chorioamnionitis (103.7 ng/mL vs. 50.0 ng/mL; p = 0.0001). The presence of both microbial invasion of the amniotic cavity and histological chorioamnionitis was associated with higher median myeloperoxidase concentration (456.0 ng/mL vs. 52.9 ng/mL; p < 0.0001). The results remained significant after adjusting for gestational age. Increased amniotic fluid myeloperoxidase in microbial invasion of the amniotic cavity and histological chorioamnionitis confirm a role of myeloperoxidase in preterm prelabor rupture of the membranes pathophysiology.

  14. Adaptation of group A Streptococcus to human amniotic fluid.

    Directory of Open Access Journals (Sweden)

    Izabela Sitkiewicz

    Full Text Available BACKGROUND: For more than 100 years, group A Streptococcus has been identified as a cause of severe and, in many cases, fatal infections of the female urogenital tract. Due to advances in hospital hygiene and the advent of antibiotics, this type of infection has been virtually eradicated. However, within the last three decades there has been an increase in severe intra- and post-partum infections attributed to GAS. METHODOLOGY: We hypothesized that GAS alters its transcriptome to survive in human amniotic fluid (AF and cause disease. To identify genes that were up or down regulated in response to growth in AF, GAS was grown in human AF or standard laboratory media (THY and samples for expression microarray analysis were collected during mid-logarithmic, late-logarithmic, and stationary growth phases. Microarray analysis was performed using a custom Affymetrix chip and normalized hybridization values derived from three biological replicates were collected at each growth point. Ratios of AF/THY above a 2-fold change and P-value <0.05 were considered significant. PRINCIPAL FINDINGS: The majority of changes in the GAS transcriptome involved down regulation of multiple adhesins and virulence factors and activation of the stress response. We observed significant changes in genes involved in the arginine deiminase pathway and in the nucleotide de novo synthesis pathway. CONCLUSIONS/SIGNIFICANCE: Our work provides new insight into how pathogenic bacteria respond to their environment to establish infection and cause disease.

  15. Atypical Amniotic Fluid Embolism Managed with a Novel Therapeutic Regimen

    Directory of Open Access Journals (Sweden)

    Shadi Rezai

    2017-01-01

    Full Text Available Amniotic fluid embolism (AFE is the second leading cause of maternal mortality in the USA with an incidence of 1 : 15,200 births. The case fatality rate and perinatal mortality associated with AFE are 13–30% and 9–44%, respectively. This rare but devastating complication can be difficult to diagnose as many of the early signs and symptoms are nonspecific. Compounding this diagnostic challenge is a lack of effective treatment regimens which to date are mostly supportive. We present the case of a 26-year-old woman who suffered from suspected AFE and was successfully treated with the novel regimen of Atropine, Ondansetron, and Ketorolac (A-OK. The authors acknowledge that this case does not meet the new criteria proposed, by Clark in 2016, but feel that it is important to share this case report, due to dramatic patient response to the provided supportive therapy presented in this case report. We hope this case report will prompt further research into this novel approach to treating AFE with Atropine, Ondansetron, and Ketorolac.

  16. Proteomic profiling of the amniotic fluid to detect inflammation, infection, and neonatal sepsis.

    Directory of Open Access Journals (Sweden)

    Catalin S Buhimschi

    2007-01-01

    Full Text Available Proteomic analysis of amniotic fluid shows the presence of biomarkers characteristic of intrauterine inflammation. We sought to validate prospectively the clinical utility of one such proteomic profile, the Mass Restricted (MR score.We enrolled 169 consecutive women with singleton pregnancies admitted with preterm labor or preterm premature rupture of membranes. All women had a clinically indicated amniocentesis to rule out intra-amniotic infection. A proteomic fingerprint (MR score was generated from fresh samples of amniotic fluid using surface-enhanced laser desorption ionization (SELDI mass spectrometry. Presence or absence of the biomarkers of the MR score was interpreted in relationship to the amniocentesis-to-delivery interval, placental inflammation, and early-onset neonatal sepsis for all neonates admitted to the Newborn Special Care Unit (n = 104. Women with "severe" amniotic fluid inflammation (MR score of 3 or 4 had shorter amniocentesis-to-delivery intervals than women with "no" (MR score of 0 inflammation or even "minimal" (MR score of 1 or 2 inflammation (median [range] MR 3-4: 0.4 d [0.0-49.6 d] versus MR 1-2: 3.8 d [0.0-151.2 d] versus MR 0: 17.0 d [0.1-94.3 d], p 100 cells/mm3, whereas the combination of Gram stain and MR score was best for rapid prediction of intra-amniotic infection (positive amniotic fluid culture.High MR scores are associated with preterm delivery, histological chorioamnionitis, and early-onset neonatal sepsis. In this study, proteomic analysis of amniotic fluid was shown to be the most accurate test for diagnosis of intra-amniotic inflammation, whereas addition of the MR score to the Gram stain provides the best combination of tests to rapidly predict infection.

  17. Amniotic fluid cells are more efficiently reprogrammed to pluripotency than adult cells.

    Science.gov (United States)

    Galende, Elisa; Karakikes, Ioannis; Edelmann, Lisa; Desnick, Robert J; Kerenyi, Thomas; Khoueiry, Georges; Lafferty, James; McGinn, Joseph T; Brodman, Michael; Fuster, Valentin; Hajjar, Roger J; Polgar, Katalin

    2010-04-01

    Recently, cultured human adult skin cells were reprogrammed to induced pluripotent stem (iPS) cells, which have characteristics similar to human embryonic stem (hES) cells. Patient-derived iPS cells offer genetic and immunologic advantages for cell and tissue replacement or engineering. The efficiency of generating human iPS cells has been very low; therefore an easily and efficiently reprogrammed cell type is highly desired. Here, we demonstrate that terminally differentiated human amniotic fluid (AF) skin cells provide an accessible source for efficiently generating abundant-induced pluripotent stem (AF-iPS) cells. By induction of pluripotency with the transcription factor quartet (OCT3/4, SOX2, KLF4, and c-MYC) the terminally differentiated, cultured AF skin cells formed iPS colonies approximately twice as fast and yielded nearly a two-hundred percent increase in number, compared to cultured adult skin cells. AF-iPS cells were identical to hES cells for morphological and growth characteristics, antigenic stem cell markers, stem cell gene expression, telomerase activity, in vitro and in vivo differentiation into the three germ layers and for their capacity to form embryoid bodies (EBs) and teratomas. Our findings provide a biological interesting conclusion that these fetal AF cells are more rapidly, easily, and efficiently reprogrammed to pluripotency than neonatal and adult cells. AF-iPS cells may have a "young," more embryonic like epigenetic background, which may facilitate and accelerate pluripotency. The ability to efficiently and rapidly reprogram terminally differentiated AF skin cells and generate induced pluripotent stem cells provides an abundant iPS cell source for various basic studies and a potential for future patient-specific personalized therapies.

  18. Amniotic fluid biochemistry in isolated polyhydramnios: a series of 464 cases.

    Science.gov (United States)

    Allaf, Bichr; Dreux, Sophie; Schmitz, Thomas; Czerkiewicz, Isabelle; Le Vaillant, Claudine; Benachi, Alexandra; Houfflin-Debarge, Véronique; Maréchaud, Martine; Oury, Jean-François; Muller, Françoise

    2015-12-01

    To report amniotic fluid biochemistry in a large series of 464 cases of isolated polyhydramnios in order to analyze both the outcome and the benefit of amniotic fluid biochemistry. This retrospective cohort (2008-2012) included polyhydramnios cases for which amniotic fluid samples were sent to our laboratory for biochemical analysis (total protein, alpha-fetoprotein and gamma-glutamyl transpeptidase) so as to investigate the etiology. A Bartter index and an esophageal atresia index were defined. Final diagnoses were compared between groups to determine the association between these indices and the frequency and type of adverse outcomes. Among 464 cases of polyhydramnios considered isolated at ultrasound examination, severe fetal diseases were diagnosed in 136 (29.3%): 46 (9.9%) chromosomal anomalies, 28 (6%) Bartter syndrome, 23 (4.95%) other genetic syndromes, 22 (4.75%) swallowing disorders and 17 (3.7%) uro-nephrological disorders. Amniotic fluid biochemistry identified esophageal atresia with 66.6% (10/15) sensitivity and 100% specificity and Bartter syndrome with 85.7% (24/28) sensitivity and 84.2% specificity. Isolated polyhydramnios is associated with a high risk of severe fetal diseases. Molecular cytogenetics and amniotic fluid biochemistry are helpful tools. © 2015 John Wiley & Sons, Ltd.

  19. 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.

  20. Amniotic fluid gamma-glutamyl transpeptidase activity during the second trimester.

    Science.gov (United States)

    Legge, M; Potter, H C

    1986-03-12

    Gamma glutamyl transpeptidase (GGTP) activity was determined in second trimester amniotic fluid taken from normal fetuses and those with fetal abnormalities. GGTP activity decreased with advancing gestation. Increasing meconium contamination correlated with an increase in GGTP activity as did increasing fetal blood contamination. Maternal blood did not affect GGTP activity. Anencephaly did not significantly alter the GGTP activity, however, fetuses with spina bifida had significantly lower activity. Klinefelters and Turners syndromes both had GGTP activity close to the 50th percentile, and two trisomy 21 fetuses had GGTP activity below the 40th percentile. Two trisomy 18 fetuses and two translocation Downs syndromes (46 XY, t (14;21) had GGTP activities considerably lower than the 20th percentile as did a fetus with gastroschisis. Second trimester amniotic fluid GGTP activity may provide an easy preliminary test to screen amniotic fluids for the possibility of certain fetal chromosome abnormalities.

  1. Sources of amniotic fluid erythropoietin during normoxia and hypoxia in fetal sheep.

    Science.gov (United States)

    Brace, Robert A; Cheung, Cecilia Y; Davis, Lowell E; Gagnon, Robert; Harding, Richard; Widness, John A

    2006-07-01

    Erythropoietin is present in human amniotic fluid and has been suggested as a marker of fetal hypoxia. The objectives of the present study were to determine whether erythropoietin is present in ovine amniotic fluid, fetal urine, and/or lung liquid and whether concentrations in these compartments change in parallel with endogenous fetal plasma erythropoietin concentration when the latter is increased experimentally. In late gestation chronically catheterized fetal sheep, samples of amniotic fluid and plasma, urine and plasma, lung liquid, amniotic fluid, and plasma were collected before and up to 7 days after induction of 4 types of fetal hypoxia: (1) acute anemic hypoxia that was induced by a single fetal hemorrhage, (2) progressive anemic hypoxia that was induced by daily exchange transfusion, (3) acute hypoxic hypoxia that was induced by the reduction of maternal inspired oxygen content, or (4) chronic placental insufficiency that was induced by daily umbilicoplacental embolization for 4 days. Erythropoietin concentrations were determined by radioimmunoassay. Statistical testing included analysis of variance and least squares regression. Under basal, nonhypoxic conditions, amniotic fluid erythropoietin concentration averaged 33.2% +/- 1.6% (SE) of fetal plasma erythropoietin concentration, and basal fetal urine and lung liquid erythropoietin concentrations ranged from low (humans, basal amniotic fluid and plasma erythropoietin concentrations were correlated only weakly (r = 0.259; r2 = 6.7%; P = .0027; n = 132). Amniotic fluid erythropoietin concentration approximately doubled after 12 hours of severe hypoxic hypoxia or after 24 hours of embolization-induced severe hypoxia but was unchanged after 12 hours of mild-moderate hypoxic hypoxia or 24 hours of anemic hypoxia. Concomitant fetal plasma erythropoietin concentrations increased to 28.1 +/- 5.3, 12.5 +/- 2.7, 10.8 +/- 4.6, and 10.0 +/- 1.3 times basal values, respectively. During progressive fetal anemia

  2. Amniotic fluid chemokines and autism spectrum disorders: An exploratory study utilizing a Danish Historic Birth Cohort

    DEFF Research Database (Denmark)

    Abdallah, Morsi; Larsen, Nanna Brink; Grove, Jakob

    2012-01-01

    Elevated levels of chemokines have been reported in plasma and brain tissue of individuals with Autism Spectrum Disorders (ASD). The aim of this study was to examine chemokine levels in amniotic fluid (AF) samples of individuals diagnosed with ASD and their controls.......Elevated levels of chemokines have been reported in plasma and brain tissue of individuals with Autism Spectrum Disorders (ASD). The aim of this study was to examine chemokine levels in amniotic fluid (AF) samples of individuals diagnosed with ASD and their controls....

  3. Amniotic fluid and colostrum as potential diets in the critical care of preterm infants

    DEFF Research Database (Denmark)

    Støy, Ann Cathrine Findal; Viberg Østergaard, Mette; Torp Sangild, Per

    2015-01-01

    Amniotic fluid is the enteral “diet” of the developing fetus, while the first mammary gland secretion, colostrum, is the natural diet of the newborn mammal. Both diets contain nutrients but also growth factors, immune-modulating components, and antibacterial agents that support perinatal organ...... be species independent, maybe allowing colostrum from one species (i.e., lactating cows) to be used as the first enteral diet for infants for whom mother’s milk is lacking. The use of amniotic fluid and bovine colostrum in the critical care of neonates is still at an experimental stage, but animal studies...

  4. Biochemical composition of amniotic fluid in normal puppies at term of pregnancy: preliminary data

    Directory of Open Access Journals (Sweden)

    Barbara Bolis

    2017-05-01

    Full Text Available The full knowledge of the normal fetal fluids composition could be useful in the dog for the better management of newborns. The aim of the present study was to define the biochemical composition of amniotic fluid of puppies born by elective Caesarean section (CS at term of pregnancy. The study enrolled 24 purebred bitches, classified into small size (20kg. All the bitches were healthy and clinically monitored from mating until parturition. For all the bitches an elective CS at term of pregnancy was performed [1]. For each puppy, the amniotic fluid was collected, immediately centrifuged and frozen at – 20° C until analysis for ALB, AMY, TB, CHOL, CK, ALP, GGT, AST, ALT, LDH, Mg, Ca, K, Na, Trig, BUN, Glc, TP, CREA, LIP, Cl, and GLOB. Data were analyzed by ANCOVA to verify the possible effects of parity, breed body size and newborn gender on amniotic biochemical composition. A total of 69 amniotic fluid samples were collected. The amniotic mean±SD and min-max values for each parameter were defined. LDH (p<0.01 and CK activity (p<0.05, as well as Glc concentrations (p<0.0001 were negatively influenced by the parity. AMY activity was significantly (p<0.05 higher in large sized (44.2±20.87 U/L respect to small sized dogs (30.3±19.89 U/L, while lower (p<0.05 CHOL amniotic concentrations were found in small sized (3.0±2.71 mg/dl as compared to large sized (3.9±2.93 mg/dl dogs. Gender of the newborn did not influence the amniotic biochemical composition. The preliminary results of this study showed some similarities as well as some differences concerning the biochemical composition of the amniotic fluid in dogs at term of pregnancy if compared to data reported for the cat [2]. Furthermore, the results suggested that, in dogs, some amniotic parameters could be influenced by breed body size and by parity.

  5. Multidimensional proteomics analysis of amniotic fluid to provide insight into the mechanisms of idiopathic preterm birth.

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    Irina A Buhimschi

    2008-04-01

    Full Text Available Though recent advancement in proteomics has provided a novel perspective on several distinct pathogenetic mechanisms leading to preterm birth (inflammation, bleeding, the etiology of most preterm births still remains elusive. We conducted a multidimensional proteomic analysis of the amniotic fluid to identify pathways related to preterm birth in the absence of inflammation or bleeding.A proteomic fingerprint was generated from fresh amniotic fluid using surface-enhanced laser desorbtion ionization time of flight (SELDI-TOF mass spectrometry in a total of 286 consecutive samples retrieved from women who presented with signs or symptoms of preterm labor or preterm premature rupture of the membranes. Inflammation and/or bleeding proteomic patterns were detected in 32% (92/286 of the SELDI tracings. In the remaining tracings, a hierarchical algorithm was applied based on descriptors quantifying similarity/dissimilarity among proteomic fingerprints. This allowed identification of a novel profile (Q-profile based on the presence of 5 SELDI peaks in the 10-12.5 kDa mass area. Women displaying the Q-profile (mean+/-SD, gestational age: 25+/-4 weeks, n = 40 were more likely to deliver preterm despite expectant management in the context of intact membranes and normal amniotic fluid clinical results. Utilizing identification-centered proteomics techniques (fluorescence two-dimensional differential gel electrophoresis, robotic tryptic digestion and mass spectrometry coupled with Protein ANalysis THrough Evolutionary Relationships (PANTHER ontological classifications, we determined that in amniotic fluids with Q-profile the differentially expressed proteins are primarily involved in non-inflammatory biological processes such as protein metabolism, signal transduction and transport.Proteomic profiling of amniotic fluid coupled with non-hierarchical bioinformatics algorithms identified a subgroup of patients at risk for preterm birth in the absence of intra-amniotic

  6. Antioxidant Vitamin Status in the Serum and Amniotic Fluid of Women with Premature Rupture of the Fetal Membranes.

    Science.gov (United States)

    Barrett, Bridget M.

    The purpose of this study was to examine the status of antioxidant vitamins in women with premature rupture of the fetal membranes. Specimens of blood and amniotic fluid were obtained from 80 pregnant subjects included both smokers and non-smokers during the third trimester. The concentrations of ascorbic acid (ASA), beta -carotene, retinol and alpha -tocopherol in serum and amniotic fluid were determined. The experimental group consisted of those subjects with PROM while the control subjects were those with normal pregnancy. No statistical differences were found between the PROM and control groups in retinol and vitamin E concentrations in amniotic fluid and serum. Serum ASA concentrations of PROM subjects were not different from controls, but the PROM subjects had significantly lower amniotic fluid ASA concentrations. However, in a study with fewer subjects a lower serum ASA concentration in the PROM subjects was observed. The ratio of amniotic fluid ASA concentration to ASA serum concentration was significantly lower in PROM patients than in controls in both studies. This suggests that low levels of ASA in the amniotic fluid, but not in serum is better associated with PROM. A low amniotic fluid concentration of ASA may reflect an inefficient transfer and/or increased fetal utilization. Alterations in ASA concentration in the amniotic fluid may affect the integrity of the chorioamnion leading to PROM. beta -Carotene was not found in the amniotic fluid. Serum beta-carotene levels were significantly lower in the PROM group compared to the control group. Low concentrations of beta-carotene in maternal serum in smokers not only associated with poor maternal outcome (PROM) but also compromised the fetal outcome (decreased birth weight). Maintenance of adequate serum beta-carotene concentration and amniotic fluid ASA in smokers may result in better maternal and fetal outcome. This study demonstrated that nutrition is an important factor in the prevention of PROM.

  7. Relationship Between Ultrasound Estimated Amniotic Fluid Index and Fetal Weight in Healthy Pregnant African Women

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    Ademola A Adeyekun

    2013-01-01

    Full Text Available Introduction: Fetal weight (FW estimation in late pregnancy is an important guide in obstetric care. Amniotic fluid protects the fetus against traumatic and infective insults. There possibly exists a relationship between FW and amniotic fluid index (AFI that can be estimated by ultrasonography. Materials and Methods: Two hundred and fifty-eight low-risk pregnancies were prospectively studied by means of ultrasound over a 12-month period. FW was estimated using a combination of fetal parameters; bi-parietal diameter, fetal trunk cross sectional area, and femur length. AFI was assessed using the 4-quadrant method. Spearman′s correlation was used to test possible relationship between amniotic fluid indices and estimated FW pairs. The level of statistical significance was set at P ≤ 0.05. Results: The mean AF1 and estimated fetal weight (EFW pairs were as follows: at 27-29 weeks the values were 172.1 mm and 1,250.2 g; at 30-32 weeks AF1 and EFW values were 170.3 mm and 1,648.0 g; at 33-35 weeks values were 162.3 mm and 2,273.5 g; at 36-38 weeks values were 144.09 mm and 2,906.1 g; at 39-40 weeks AF1 and EFW values were 125.0 mm and 3,222.6 g. Overall, there was no statistically significant relationship between AFI and EFW ( P > 0.05; r = 0.241. Conclusion: While FW calculations and amniotic index showed variations in value in late pregnancy, there does not appear to be a linear relationship between ultrasound estimate of FW and amniotic index. The implication of this is that fetal size need not be taken into cognizance when alterations in amniotic fluid values are noted.

  8. What Is New in Amniotic Fluid Embolism?: Best Articles From the Past Year.

    Science.gov (United States)

    Benson, Michael D

    2017-05-01

    This month, we focus on current research in amniotic fluid embolism. Dr. Benson discusses four recent publications, and each is concluded with a "bottom line" that is the take-home message. The complete reference for each can be found in on this page, along with direct links to the abstracts.

  9. Concentration of amoxicillin in maternal serum, cord blood, amniotic fluid and the placenta after vaginal administration.

    Science.gov (United States)

    Zaręba-Szczudlik, Julia; Romejko-Wolniewicz, Ewa; Lewandowski, Zbigniew; Różańska, Hanna; Czajkowski, Krzysztof

    2015-01-01

    The aim of this study was to assess the amoxicillin concentration in maternal serum, cord blood, amniotic fluid and the placenta, 2 h following vaginal administration and the factors influencing the drug level. Twenty-eight full-term pregnant women who qualified for elective cesarean delivery were included in the study. Vaginal suppositories containing 250 mg of amoxicillin were administered 2 h prior to the operation. Amoxicillin levels were determined using the diffusion microbial assay. The amoxicillin level in amniotic fluid was significantly higher in comparison to that of maternal serum, cord blood or the placenta. Maternal age positively and gestational weight gain negatively correlated with the amoxicillin concentration in maternal serum. The maternal serum hemoglobin level and red blood cell count were positively correlated with amoxicillin concentration in the amniotic fluid. Neonatal birth weight was positively correlated with maternal serum and cord blood amoxicillin levels. Hypertensive women had significantly higher amoxicillin concentrations in amniotic fluid, and women with thrombocytopenia presented significantly higher cord blood amoxicillin concentrations. Amoxicillin presented poor concentration in maternal-fetal compartments after vaginal administration, but the factors influencing the drug level in different compartments require further investigation.

  10. Amniotic fluid RNA gene expression profiling provides insights into the phenotype of Turner syndrome.

    Science.gov (United States)

    Massingham, Lauren J; Johnson, Kirby L; Scholl, Thomas M; Slonim, Donna K; Wick, Heather C; Bianchi, Diana W

    2014-09-01

    Turner syndrome is a sex chromosome aneuploidy with characteristic malformations. Amniotic fluid, a complex biological material, could contribute to the understanding of Turner syndrome pathogenesis. In this pilot study, global gene expression analysis of cell-free RNA in amniotic fluid supernatant was utilized to identify specific genes/organ systems that may play a role in Turner syndrome pathophysiology. Cell-free RNA from amniotic fluid of five mid-trimester Turner syndrome fetuses and five euploid female fetuses matched for gestational age was extracted, amplified, and hybridized onto Affymetrix(®) U133 Plus 2.0 arrays. Significantly differentially regulated genes were identified using paired t tests. Biological interpretation was performed using Ingenuity Pathway Analysis and BioGPS gene expression atlas. There were 470 statistically significantly differentially expressed genes identified. They were widely distributed across the genome. XIST was significantly down-regulated (p Turner syndrome transcriptome from other aneuploidies we previously studied. Manual curation of the differentially expressed gene list identified genes of possible pathologic significance, including NFATC3, IGFBP5, and LDLR. Transcriptomic differences in the amniotic fluid of Turner syndrome fetuses are due to genome-wide dysregulation. The hematologic/immune system differences may play a role in early-onset autoimmune dysfunction. Other genes identified with possible pathologic significance are associated with cardiac and skeletal systems, which are known to be affected in females with Turner syndrome. The discovery-driven approach described here may be useful in elucidating novel mechanisms of disease in Turner syndrome.

  11. Stable isotope dilution analysis of orotic acid and uracil in amniotic fluid

    NARCIS (Netherlands)

    Jakobs, C.; Sweetman, L.; Nyhan, W.L.; Gruenke, L.; Craig, J.C.; Wadman, S.K.

    1984-01-01

    Rapid, sensitive and accurate stable isotope dilution assays were developed for the measurement of orotic acid and uracil in amniotic fluid. The method utilizes [15N2]orotic acid and [15N2]uracil as internal standards, isolation by liquid partition chromatography and quantitation by chemical

  12. Peptidomic Analysis of Amniotic Fluid for Identification of Putative Bioactive Peptides in Ventricular Septal Defect

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    Xing Li

    2016-05-01

    Full Text Available Background: Ventricular septal defect (VSD is one of the most common congenital heart diseases and to date the role of peptides in human amniotic fluid in the pathogenesis of VSD have been rarely investigated. Methods: To gain insight into the mechanisms of protein and peptides in cardiovascular development, we constructed a comparative peptidomic profiling of human amniotic fluid between normal and VSD fetuses using a stable isobaric labeling strategy involving tandem mass tag reagents, followed by nano liquid chromatography tandem mass spectrometry. Results: We identified and quantified 692 non-redundant peptides, 183 of which were differentially expressed in the amniotic fluid of healthy and VSD fetuses; 69 peptides were up regulated and 114 peptides were down regulated. These peptides were imported into the Ingenuity Pathway Analysis (IPA and identified putative roles in cardiovascular system morphogenesis and cardiogenesis. Conclusion: We concluded that 35 peptides located within the functional domains of their precursor proteins could be candidate bioactive peptides for VSD. The identified peptide changes in amniotic fluid of VSD fetuses may advance our current understanding of congenital heart disease and these peptides may be involved in the etiology of VSD.

  13. Peptidomic Analysis of Amniotic Fluid for Identification of Putative Bioactive Peptides in Ventricular Septal Defect.

    Science.gov (United States)

    Li, Xing; Wu, Li-Jie; Gu, Meng; Chen, Yu-Mei; Zhang, Qi-Jun; Li, Hua; Cheng, Zi-Jie; Hu, Ping; Han, Shu-Ping; Yu, Zhang-Bin; Qian, Ling-Mei

    2016-01-01

    Ventricular septal defect (VSD) is one of the most common congenital heart diseases and to date the role of peptides in human amniotic fluid in the pathogenesis of VSD have been rarely investigated. To gain insight into the mechanisms of protein and peptides in cardiovascular development, we constructed a comparative peptidomic profiling of human amniotic fluid between normal and VSD fetuses using a stable isobaric labeling strategy involving tandem mass tag reagents, followed by nano liquid chromatography tandem mass spectrometry. We identified and quantified 692 non-redundant peptides, 183 of which were differentially expressed in the amniotic fluid of healthy and VSD fetuses; 69 peptides were up regulated and 114 peptides were down regulated. These peptides were imported into the Ingenuity Pathway Analysis (IPA) and identified putative roles in cardiovascular system morphogenesis and cardiogenesis. We concluded that 35 peptides located within the functional domains of their precursor proteins could be candidate bioactive peptides for VSD. The identified peptide changes in amniotic fluid of VSD fetuses may advance our current understanding of congenital heart disease and these peptides may be involved in the etiology of VSD. © 2016 The Author(s) Published by S. Karger AG, Basel.

  14. Detection of otoacoustic emissions in chinchilla when the middle ear contains amniotic fluid.

    Science.gov (United States)

    Akinpelu, Olubunmi V; Funnell, W Robert J; Daniel, Sam J

    2015-04-01

    Otoacoustic emissions have frequently been used for newborn hearing screening. However, they have low specificities and high referral rates. The presence of amniotic fluid in the middle ear is one reason for these problems. The aim of this study was to determine the effects of human amniotic fluid on otoacoustic emissions and on middle-ear function. Forty-six chinchillas were randomly divided into eight groups based on the type (amniotic fluid or normal saline) and volume (0.5, 1, 1.5, 2 ml) of liquid introduced into the middle ear. Distortion product otoacoustic emission (DPOAE) and wideband reflectance (WBR) measurements were taken under inhalational anaesthesia before and after introduction of middle-ear liquid. The differences in these measurements were subjected to statistical analyses. Prospective controlled animal study. Significant reductions of DPOAE levels and increases in WBR occurred across all frequencies when there was liquid in the middle ear, and the changes became greater for increased volumes of liquid. Changes in the noise level had important effects on the otoacoustic-emission signal-to-noise ratio at the three lowest frequencies. Both human amniotic fluid and saline in the chinchilla middle ear resulted in changes in otoacoustic-emission detection patterns and WBR that may be relevant to newborn hearing screening. © 2014 The American Laryngological, Rhinological and Otological Society, Inc.

  15. [Lamellar body count in amniotic fluid for assessing fetal lung maturity].

    Science.gov (United States)

    Visnjevac, Jovana; Novakov-Mikić, Aleksandra; Nikolić, Aleksandra; Visnjevac, Nemanja

    2010-01-01

    Respiratory distress syndrome (RDS) of the newborn infant caused by immaturity of fetal lung is a very serious clinical problem. Surfactant is stored in the form of lamellar bodies. They are secreted into alveolar space and passed into amniotic fluid where they can be found. The similarity of lamellar body size to platelet size permits the use of a standard automated hematologic cell counter to estimate the number of lamellar bodies in amniotic fluid. We conducted a prospective clinical study from 2005-2006 on amniotic fluid samples. Amniotic fluid samples were collected near delivery by transvaginal amniotomy, amniotomy during Cesarean section and 72 hours before delivery by amniocentesis. A hematology analyzer (Nikon-Kohden) was used to determine the lamellar body counts. After birth of newborns we compared their complete clinical examination results particularly emphasizing the prediction of the method of RDS by lamellar body count. Maximally specific lamellar body cutoffs for maturity and immaturity were determined using ROC curves. Of 232 amniotic fluid samples which were tested, 112 samples were collected by transvaginal amniotomy, 88 were taken during Cesarean delivery and 32 samples were collected by amniocentesis. The incidence of RDS was 14.6%. ROC curves were used to identify cut points for the test. We found that LBC is a good screening test for predicting fetal lung maturity with the area under the curve of 0.751. LBC cutoff of 42 x 10(3)/microl, with sensitivity of 82.4% and specificity of 64.6%, proved best for predicting fetal lung maturity. LBC is a good screening test for predicting fetal lung maturity. The advantages of LBC are speed, objectivity, low price, low sample volume required and universal availability.

  16. Prevalence of human papillomavirus and herpes simplex virus in amniotic fluid from pregnant women of Eastern Poland

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    Mariusz Skoczyński

    2015-12-01

    Full Text Available Background: The exact route of human papillomavirus (HPV and herpes simplex virus (HSV transmission from a pregnant woman to her fetus has not been clearly established thus far. The data of many studies raise the possibility of intrauterine infection. In order to clarify our knowledge about virus vertical transmission in pregnant women, viral prevalence in amniotic fluid cannot. To the best of our knowledge, this is the first study on HPV DNA and HSV DNA detection in amniotic fluid in Poland.Material and methods: The study covered 138 samples of amniotic fluid from patients undergoing invasive prenatal diagnostic procedures (for medical indications during the second trimester of gestation. The aim of the study was to assess the prevalence of HPV and HSV in the amniotic fluid samples obtained from asymptomatic women with intact amniotic membranes. To identify viral DNA of HPV and HSV in collected material, polymerase chain reaction (PCR was performed.Results: We did not find HPV or HSV DNA in any of the examined specimens of amniotic fluid.Conclusion: Our investigation did not confirm the prenatal transmission of HPV and HSV to the amniotic fluid.

  17. Committee Opinion No.689 Summary: Delivery of a Newborn With Meconium-Stained Amniotic Fluid.

    Science.gov (United States)

    2017-03-01

    In 2006, the American Academy of Pediatrics and the American Heart Association published the 2005 guidelines on neonatal resuscitation. Before the 2005 guidelines, management of a newborn with meconium-stained amniotic fluid included suctioning of the oropharynx and nasopharynx on the perineum after the delivery of the head but before the delivery of the shoulders. The 2005 guidelines did not support this practice because routine intrapartum suctioning does not prevent or alter the course of meconium aspiration syndrome in vigorous newborns. However, the 2005 guidelines did support intubation of the trachea and suctioning of meconium or other aspirated material from beneath the glottis in nonvigorous newborns. In 2015, the guidelines were updated. Routine intubation and tracheal suctioning are no longer required. If the infant is vigorous with good respiratory effort and muscle tone, the infant may stay with the mother to receive the initial steps of newborn care. If the infant born through meconium-stained amniotic fluid presents with poor muscle tone and inadequate breathing efforts, the initial steps of resuscitation should be completed under the radiant warmer. Appropriate intervention to support ventilation and oxygenation should be initiated as indicated for each infant. Infants with meconium-stained amniotic fluid should no longer routinely receive intrapartum suctioning, whether they are vigorous or not. In addition, meconium-stained amniotic fluid is a condition that requires the notification and availability of an appropriately credentialed team with full resuscitation skills, including endotracheal intubation. Resuscitation should follow the same principles for infants with meconium-stained fluid as for those with clear fluid.

  18. Committee Opinion No 689: Delivery of a Newborn With Meconium-Stained Amniotic Fluid.

    Science.gov (United States)

    2017-03-01

    In 2006, the American Academy of Pediatrics and the American Heart Association published the 2005 guidelines on neonatal resuscitation. Before the 2005 guidelines, management of a newborn with meconium-stained amniotic fluid included suctioning of the oropharynx and nasopharynx on the perineum after the delivery of the head but before the delivery of the shoulders. The 2005 guidelines did not support this practice because routine intrapartum suctioning does not prevent or alter the course of meconium aspiration syndrome in vigorous newborns. However, the 2005 guidelines did support intubation of the trachea and suctioning of meconium or other aspirated material from beneath the glottis in nonvigorous newborns. In 2015, the guidelines were updated. Routine intubation and tracheal suctioning are no longer required. If the infant is vigorous with good respiratory effort and muscle tone, the infant may stay with the mother to receive the initial steps of newborn care. If the infant born through meconium-stained amniotic fluid presents with poor muscle tone and inadequate breathing efforts, the initial steps of resuscitation should be completed under the radiant warmer. Appropriate intervention to support ventilation and oxygenation should be initiated as indicated for each infant. Infants with meconium-stained amniotic fluid should no longer routinely receive intrapartum suctioning, whether they are vigorous or not. In addition, meconium-stained amniotic fluid is a condition that requires the notification and availability of an appropriately credentialed team with full resuscitation skills, including endotracheal intubation. Resuscitation should follow the same principles for infants with meconium-stained fluid as for those with clear fluid.

  19. Transport-associated pathway responses in ovine fetal membranes to changes in amniotic fluid dynamics.

    Science.gov (United States)

    Cheung, Cecilia Y; Anderson, Debra F; Brace, Robert A

    2017-11-01

    Current evidence suggests that amniotic fluid volume (AFV) is actively regulated by vesicular transport of amniotic fluid outward across the amnion and into the underlying fetal vasculature in the placenta. Our objective was to determine whether gene expression profiles of potential stimulators, inhibitors, and mediators of vesicular transport are altered in response to changes in intramembranous absorption (IMA) rate. Samples of ovine amnion and chorion were obtained from fetal sheep with normal, experimentally reduced or increased AFVs and IMA rates. Amnion and chorion levels of target mRNAs were determined by RT-qPCR In the amnion, caveolin-1 and flotillin-1 mRNA levels were unchanged during alterations in IMA rate. However, levels of both were significantly higher in amnion than in chorion. Tubulin- α mRNA levels in the amnion but not in chorion were reduced when IMA rate decreased, and amnion levels correlated positively with IMA rate ( P  fluid and thus modulate IMA rate. Further, our results are consistent with the concept that the amnion is the rate-limiting layer for amniotic fluid transport. © 2017 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.

  20. [Imbalance of system of glutamin - glutamic acid in the placenta and amniotic fluid at placental insufficiency].

    Science.gov (United States)

    Pogorelova, T N; Gunko, V O; Linde, V A

    2014-01-01

    Metabolism of glutamine and glutamic acid has been investigated in the placenta and amniotic fluid under conditions of placental insufficiency. The development of placental insufficiency is characterized by the increased content of glutamic acid and a decrease of glutamine in both placenta and amniotic fluid. These changes changes were accompanied by changes in the activity of enzymes involved in the metabolism of these amino acids. There was a decrease in glutamate dehydrogenase activity and an increase in glutaminase activity with the simultaneous decrease of glutamine synthetase activity. The compensatory decrease in the activity of glutamine keto acid aminotransferase did not prevent a decrease in the glutamine level. The impairments in the system glutamic acid-glutamine were more pronounced during the development of premature labor.

  1. Metal-based particles in human amniotic fluids of fetuses with normal karyotype and congenital malformation--a pilot study.

    Science.gov (United States)

    Barošová, H; Dvořáčková, J; Motyka, O; Kutláková, K Mamulová; Peikertová, P; Rak, J; Bielniková, H; Kukutschová, J

    2015-05-01

    This study explores the inorganic composition of amniotic fluid in healthy human fetuses and fetuses with congenital malformation with a special attention to presence of metal-based solid particles. Amniotic fluid originates from maternal blood and provides fetus mechanical protection and nutrients. In spite of this crucial role, the environmental impact on the composition of amniotic fluid remains poorly studied. The samples of human amniotic fluids were obtained by amniocentesis, including both healthy pregnancies and those with congenital malformations. The samples were analysed using several techniques, including Raman microspectroscopy, scanning electron microscopy with energy-dispersed spectrometry (SEM-EDS), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis. Several metal-based particles containing barium, titanium, iron, and other elements were detected by SEM-EDS and Raman microspectroscopy. XRD analysis detected only sodium chloride as the main component of all amniotic fluid samples. Infrared spectroscopy detected protein-like organic components. Majority of particles were in form of agglomerates up to tens of micrometres in size, consisting of mainly submicron particles. By statistical analysis (multiple correspondence analysis), it was observed that groups of healthy and diagnosed fetuses form two separate groups and therefore, qualitative differences in chemical composition may have distinct biological impact. Overall, our results suggest that metal-based nanosized pollutants penetrate into the amniotic fluid and may affect human fetuses.

  2. [Determination of tramadol and its active metabolite O-desmethyltramadol in plasma and amniotic fluid using LC/MS/MS].

    Science.gov (United States)

    Zhao, Li-mei; Chen, Xiao-yan; Cui, Jian-jun; Sunita, Maleku; Zhong, Da-fang

    2004-06-01

    To determinate tramadol and O-desmethyltramadol in human plasma and amniotic fluid by LC/MS/MS, and distribution of tramadol and O-desmethyltramadol in maternity and fetus were studied. Samples containing tramadol, O-desmethyltramadol and diphenhydramine (internal standard, IS ) were extracted using liquid-liquid extraction, followed by liquid chromatographic separation and on-line MS/MS using atmospheric pressure chemical ionization as an interface detection. The analytes were detected in the selected reaction monitoring mode. The calibration curves for tramadol and O-desmethytramadol in plasma and amniotic fluid were linear in the range from 8.0 to 800.0 microg x L(-1) (plasma) and 1.0 to 400.0 microg x L(-1) (amniotic fluid). The method was applied to the measurement of tramadol and O-desmethytramadol concentrations in maternal vein, umbilical vein, umbilical artery and amniotic fluid. Following intramuscular pre-operative administration 1.5 mg x kg(-1) doses of tramadol to parturients, plasma concentrations of tramadol were significantly higher than those in amniotic fluid. The concentrations of O-desmethyltramadol in plasma were lower, and were not detected in amniotic fluid. The method is shown to be accurate, robust and convenient, and suitable for clinical pharmacokinetics studies of tramadol and O-desmethyltramadol.

  3. Accuracy of real-time polymerase chain reaction for Toxoplasma gondii in amniotic fluid.

    Science.gov (United States)

    Wallon, Martine; Franck, Jacqueline; Thulliez, Philippe; Huissoud, Cyril; Peyron, François; Garcia-Meric, Patricia; Kieffer, François

    2010-04-01

    To provide clinicians with information about the accuracy of real-time polymerase chain reaction (PCR) analysis of amniotic fluid for the prenatal diagnosis of congenital Toxoplasma infection. This was a prospective cohort study of women with Toxoplasma infection identified by prenatal screening in three centers routinely carrying out real-time PCR for the detection of Toxoplasma gondii in amniotic fluid. The data available were gestational age at maternal infection, types and dates of maternal treatment, results of amniocentesis and neonatal work-up and definitive infectious status of the child. We estimated sensitivity, specificity and positive and negative predictive values both overall and per trimester of pregnancy at the time of maternal infection. Polymerase chain reaction analysis was carried out on amniotic fluid for 261 of the 377 patients included (69%). It was accurate with the exception of four negative results in children who were infected. Overall sensitivity and negative predictive value were 92.2% (95% confidence interval [CI] 81-98%) and 98.1% (95% CI 95-99.5%), respectively. There was no significant association with the trimester of pregnancy during which maternal infection occurred. Specificity and positive predictive values of 100% were obtained for all trimesters. Real-time PCR analysis significantly improves the detection of T. gondii on amniotic fluid. It provides an accurate tool to predict fetal infection and to decide on appropriate treatment and surveillance. However, postnatal follow-up remains necessary in the first year of life to fully exclude infection in children for whom PCR results were negative. III.

  4. RESPIRATORY PROBLEMS IN NEWBORNS ASSOCIATED WITH MECONIUM-STAINED AMNIOTIC FLUID

    OpenAIRE

    Sudha Menon

    2017-01-01

    BACKGROUND Perinatal morbidity and mortality resulting from aspiration of Meconium-Stained Amniotic Fluid (MSAF) is due to the respiratory problems ranging from mild respiratory distress to meconium aspiration syndrome and meconium pneumonitis. The aim of the study is to define the respiratory problems and to identify the determinants of respiratory distress in babies born through light, moderate and thick MSAF. MATERIALS AND METHODS This was a prospective observational st...

  5. [Cultivation of amniotic fluid cells for the purpose of diagnosing certain metabolic diseases].

    Science.gov (United States)

    Zolotukhina, T V; Tsvetkova, I V

    1980-03-01

    A method for preparing amniotic fluid cell culture has been developed. The optimal conditions for the culture involved the incubation period averaging 17.9 days. Normal activity values were obtained for 8 glycosidases whose deficiency is responsible for the development of hereditary accumulation diseases. The data obtained allow a prenatal diagnosis of glycolipidoses (Tay-Sachs, Fabrys, Gaucher's diseases, etc.). The evidence on prenatal diagnosis of Tay-Sach's disease is presented.

  6. Borderline amniotic fluid index and perinatal outcomes in the uncomplicated term pregnancy.

    Science.gov (United States)

    Choi, Soo Ran

    2016-01-01

    To determine perinatal outcomes in uncomplicated term pregnancies with a borderline amniotic fluid index (AFI). A retrospective review was conducted of uncomplicated singleton pregnancies at term (>37 weeks). Borderline and normal AFI were defined as 5.1 ≤ AFI ≤ 8.0 cm and 8.1 ≤ AFI ≤ 24 cm, respectively. Adverse perinatal outcomes, cesarean delivery for non-reassuring fetal heart rate testing, meconium-stained amniotic fluid, a 5-min Apgar score of borderline and normal AFI groups. Borderline AFI was not significantly associated with cesarean delivery for non-reassuring fetal heart rate testing (p = 0.513), meconium-stained amniotic fluid (p = 0.641), admission to the NICU (p = 0.368), or a 5-min Apgar score of borderline group. Multiple logistic regression analysis showed that borderline AFI was not associated with cesarean delivery for non-reassuring fetal heart rate testing (odds ratio [OR] = 0.72, 95% confidence interval [CI] 0.27-1.91, p = 0.52). In uncomplicated term pregnancies, a borderline AFI does not increase the risk of adverse perinatal outcomes.

  7. Amniotic fluid volume: Rapid MR-based assessment at 28-32 weeks gestation

    Energy Technology Data Exchange (ETDEWEB)

    Hilliard, N.J.; Hawkes, R.; Patterson, A.J.; Graves, M.J.; Priest, A.N.; Hunter, S.; Set, P.A.; Lomas, D.J. [Cambridge University Hospitals NHS Foundation Trust, Department of Radiology, Cambridge (United Kingdom); Lees, C. [Imperial College Healthcare NHS Trust, Department of Obstetrics and Fetal Medicine, London (United Kingdom)

    2016-10-15

    This work evaluates rapid magnetic resonance projection hydrography (PH) based amniotic fluid volume (AFV) estimates against established routine ultrasound single deepest vertical pocket (SDVP) and amniotic fluid index (AFI) measurements, in utero at 28-32 weeks gestation. Manual multi-section planimetry (MSP) based measurement of AFV is used as a proxy reference standard. Thirty-five women with a healthy singleton pregnancy (20-41 years) attending routine antenatal ultrasound were recruited. SDVP and AFI were measured using ultrasound, with same day MRI assessing AFV with PH and MSP. The relationships between the respective techniques were assessed using linear regression analysis and Bland-Altman method comparison statistics. When comparing estimated AFV, a highly significant relationship was observed between PH and the reference standard MSP (R{sup 2} = 0.802, p < 0.001). For the US measurements, SDVP measurement related most closely to amniotic fluid volume, (R{sup 2} = 0.470, p < 0.001), with AFI demonstrating a weaker relationship (R{sup 2} = 0.208, p = 0.007). This study shows that rapid MRI based PH measurement is a better predictor of AFV, relating more closely to our proxy standard than established US techniques. Although larger validation studies across a range of gestational ages are required this approach could form part of MR fetal assessment, particularly where poly- or oligohydramnios is suspected. (orig.)

  8. Evaluation of two real time PCR assays for the detection of bacterial DNA in amniotic fluid.

    Science.gov (United States)

    Girón de Velasco-Sada, Patricia; Falces-Romero, Iker; Quiles-Melero, Inmaculada; García-Perea, Adela; Mingorance, Jesús

    2018-01-01

    The aim of this study was to evaluate two non-commercial Real-Time PCR assays for the detection of microorganisms in amniotic fluid followed by identification by pyrosequencing. We collected 126 amniotic fluids from 2010 to 2015 for the evaluation of two Real-Time PCR assays for detection of bacterial DNA in amniotic fluid (16S Universal PCR and Ureaplasma spp. specific PCR). The method was developed in the Department of Microbiology of the University Hospital La Paz. Thirty-seven samples (29.3%) were positive by PCR/pyrosequencing and/or culture, 4 of them were mixed cultures with Ureaplasma urealyticum. The Universal 16S Real-Time PCR was compared with the standard culture (81.8% sensitivity, 97.4% specificity, 75% positive predictive value, 98% negative predictive value). The Ureaplasma spp. specific Real-Time PCR was compared with the Ureaplasma/Mycoplasma specific culture (92.3% sensitivity, 89.4% specificity, 50% positive predictive value, 99% negative predictive value) with statistically significant difference (p=0.005). Ureaplasma spp. PCR shows a rapid response time (5h from DNA extraction until pyrosequencing) when comparing with culture (48h). So, the response time of bacteriological diagnosis in suspected chorioamnionitis is reduced. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Maternal Plasma and Amniotic Fluid Chemokines Screening in Fetal Down Syndrome

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    Piotr Laudanski

    2014-01-01

    Full Text Available Objective. Chemokines exert different inflammatory responses which can potentially be related to certain fetal chromosomal abnormalities. The aim of the study was to determine the concentration of selected chemokines in plasma and amniotic fluid of women with fetal Down syndrome. Method. Out of 171 amniocentesis, we had 7 patients with confirmed fetal Down syndrome (15th–18th weeks of gestation. For the purpose of our control, we chose 14 women without confirmed chromosomal aberration. To assess the concentration of chemokines in the blood plasma and amniotic fluid, we used a protein macroarray, which allows the simultaneous determination of 40 chemokines per sample. Results. We showed significant decrease in the concentration of 4 chemokines, HCC-4, IL-28A, IL-31, and MCP-2, and increase in the concentration of CXCL7 (NAP-2 in plasma of women with fetal Down syndrome. Furthermore, we showed decrease in concentration of 3 chemokines, ITAC, MCP-3, MIF, and increase in concentration of 4 chemokines, IP-10, MPIF-1, CXCL7, and 6Ckine, in amniotic fluid of women with fetal Down syndrome. Conclusion. On the basis of our findings, our hypothesis is that the chemokines may play role in the pathogenesis of Down syndrome. Defining their potential as biochemical markers of Down syndrome requires further investigation on larger group of patients.

  10. Amniotic fluid soluble Toll-like receptor 4 in pregnancies complicated by preterm prelabor rupture of the membranes.

    Science.gov (United States)

    Kacerovsky, Marian; Andrys, Ctirad; Hornychova, Helena; Pliskova, Lenka; Lancz, Kinga; Musilova, Ivana; Drahosova, Marcela; Bolehovska, Radka; Tambor, Vojtech; Jacobsson, Bo

    2012-07-01

    To determine amniotic fluid soluble Toll-like receptor 4 (sTLR4) levels in women with preterm prelabor rupture of the membranes according to the presence of microbial invasion of the amniotic cavity and histological chorioamnionitis and its relation to neonatal outcome. One hundred two women with singleton pregnancies with a gestational age between 24 + 0 and 36 + 6 weeks were included in a prospective cohort study. Amniocenteses were performed, and the concentrations of sTLR4 in the amniotic fluid were determined using sandwich enzyme-linked immunosorbent assay technique. Women with the presence of microbial invasion of the amniotic cavity had higher sTLR4 levels [median 54.2 ng/mL, interquartile range (IQR) 10.15-289.9] than those without this condition (median 18.1 ng/mL, IQR 8.1-29.9; p = 0.001). Women with the presence of histological chorioamnionitis had a higher sTLR4 level (median 28.0 ng/mL, IQR 11.15-178.1) compared with women without histological chorioamnionitis (median 13.0 ng/mL, IQR 7.8-28.7; p = 0.003). A mixed linear model was used to adjust for confounders. The difference was found only between women with and without microbial invasion of the amniotic cavity in this model. Microbial invasion of the amniotic cavity was associated with higher amniotic fluid sTLR4 levels independent of confounders.

  11. RESPIRATORY PROBLEMS IN NEWBORNS ASSOCIATED WITH MECONIUM-STAINED AMNIOTIC FLUID

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    Sudha Menon

    2017-04-01

    Full Text Available BACKGROUND Perinatal morbidity and mortality resulting from aspiration of Meconium-Stained Amniotic Fluid (MSAF is due to the respiratory problems ranging from mild respiratory distress to meconium aspiration syndrome and meconium pneumonitis. The aim of the study is to define the respiratory problems and to identify the determinants of respiratory distress in babies born through light, moderate and thick MSAF. MATERIALS AND METHODS This was a prospective observational study conducted in a university tertiary maternity care institute in Kerala. 150 term pregnancies with meconium-stained amniotic fluid were selected of which 50 cases were each of light, moderate and thick meconium staining. RESULTS Meconium-stained amniotic fluid was seen frequently in the age group between 20-25 years age than above age of 25 years (p<0.0001. Post-dated pregnancies and pregnancies complicated by pregnancy-induced hypertension and anaemia also showed a trend in increase occurrence of meconium-stained liquor. Severe birth asphyxia as indicated by Apgar score of <3 was seen in 72.7% of thick meconium staining compared to 18.2% and 9.1% with moderate and light staining of meconium (P=0.004. Respiratory distress in newborn was severe (meconium aspiration syndrome in babies with thick meconium staining (80% compared to 15% with moderate meconium and 5% with light meconium staining (P<0.0001. The hazard ratio for death was 8 times higher with meconium aspiration syndrome compared to newborns with aspiration of moderate or light meconium-stained amniotic fluid aspiration. The odds ratio was also very high with aspiration of thick meconium-stained amniotic fluid than the other groups (OR 11.43; 95%, CI 1.33 to 98.35; Z statistic 2.22; P=0.03. CONCLUSION Meconium staining of the liquor is an important warning signal of foetal distress and the likelihood is increased if associated with alterations in the foetal heart rate. Increased morbidity and mortality was found with

  12. Polyhydramnios in Lrp4 knockout mice with bilateral kidney agenesis: Defects in the pathways of amniotic fluid clearance.

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    Tanahashi, Hiroshi; Tian, Qing-Bao; Hara, Yoshinobu; Sakagami, Hiroyuki; Endo, Shogo; Suzuki, Tatsuo

    2016-02-05

    Amniotic fluid volume during mid-to-late gestation depends mainly on the urine excretion from the foetal kidneys and partly on the fluid secretion from the foetal lungs during foetal breathing-like movements. Urine is necessary for foetal breathing-like movements, which is critical for foetal lung development. Bilateral renal agenesis and/or obstruction of the urinary tract lead to oligohydramnios, which causes infant death within a short period after birth due to pulmonary hypoplasia. Lrp4, which functions as an agrin receptor, is essential for the formation of neuromuscular junctions. Herein, we report novel phenotypes of Lrp4 knockout (Lrp4(-/-)) mice. Most Lrp4(-/-) foetuses showed unilateral or bilateral kidney agenesis, and Lrp4 knockout resulted in polyhydramnios. The loss of Lrp4 compromised foetal swallowing and breathing-like movements and downregulated the expression of aquaporin-9 in the foetal membrane and aquaporin-1 in the placenta, which possibly affected the amniotic fluid clearance. These results suggest that amniotic fluid removal was compromised in Lrp4(-/-) foetuses, resulting in polyhydramnios despite the impairment of urine production. Our findings indicate that amniotic fluid removal plays an essential role in regulating the amniotic fluid volume.

  13. Cryopreservation of human limbal stem cells ex vivo expanded on amniotic membrane.

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    Yeh, Hui-Jung; Yao, Chao-Ling; Chen, Hsin-I; Cheng, Huey-Chuan; Hwang, Shiaw-Min

    2008-04-01

    After cornea transplantation, the donor's limbal zone is currently discarded as medical waste. However, the limbal zone is rich in limbal stem cells and can be used in therapeutic applications of limbus loss. This study aimed to increase the availability of limbal stem cells and develop the optimal conditions of cryopreservation for ex vivo expanded limbal stem cells. Pieces of the limbus were cultured on amniotic membrane (AM) to outgrow limbal stem cells as cell sheets for 3 weeks. Different formulas of cryoprotectants were tested to preserve the expanded cell sheets in liquid nitrogen. Before and after cryopreservation, expanded cell sheets were assessed for cellular characteristics by viability, histologic examination, and expression of ABCG2, vimentin, and keratin 3. Expanded cell sheets usually exhibited 3-6 stratified layers after 3-week culture on AM and expressed specific markers of ABCG2 and vimentin for limbal stem cells. The effects of cryopreservation with different cryoprotectants were analyzed by histopathology, stem cell markers, and cell viability. The results showed that the optimal formula of cryoprotectants for expanded limbal cell sheets was 60% Dulbecco modified Eagle medium, 30% fetal bovine serum, and 10% dimethyl sulfoxide. After 8-week cryopreservation in liquid nitrogen, the characteristics of limbal stem cells were maintained, and the average viability of thawed cells was 53.8% +/- 5.8%. These results showed that limbal stem cells expanded on AM could be cryopreserved and provide a promising source without delay, if banking, for patients with limbal stem cell deficiency in the future.

  14. Isolamento, caracterização e diferenciação de células-tronco mesenquimais do líquido amniótico equino obtido em diferentes idades gestacionais Isolation, characterization and differentiation of mesenchymal stem cells derived from equine amniotic fluid obtained from different gestacional ages

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    Bruna De Vita

    2013-04-01

    potencial de diferenciação em linhagens extra mesodermais já relatados na literatura.The interest in stem cells derived from fetal annexes of many species has exponentially increased during the last decades, because they are adult stem cell sources with potential of differentiation in several cell lineages; which present little or no immunogenicity and are an alternative with great importance for storage cell banks. Despite the rising interest, studies for the equine species are still rare. The aim of this study was to isolate, characterize and differentiate mesenchymal stem cells derived from equine amniotic fluid obtained from initial, middle and late third of gestation (AF-MSCs, and compare their results. Twenty three samples from equine amniotic fluid were evaluated by morphological, immunocytochemical and immunophenotypical (Flow cytometer assays and osteogenic, adipogenic and chondrogenic in vitro differentiation. All samples demonstrated plastic adhesion and fibroblastoid morphology. The immunocytochemical assay demonstrated cells from all the studied groups were positive for CD44, PCNA and vimentin and negative for cytokeratin and Oct-4. Flow cytometry demonstrated expression of CD44 and CD90 and no expression of CD34, where CD44 and CD90 markers presented decreasing pattern of expression in relation to the gestational development. All samples collected from all gestational phases were capable to differentiate in osteogenic, chondrogenic and adipogenic lineages. Thus, cells obtained from equine amniotic fluid presented morphological and immunophenotypical characteristics and potential of differentiation typical of MSCs showing that the collection can be performed at any stage of pregnancy. However, more studies should be performed about the expression of pluripotent markers as Oct-4 and the differentiation potential for extra mesodermal lineages prior demonstrated in the literature.

  15. Intraamniotic inflammatory response to bacteria: analysis of multiple amniotic fluid proteins in women with preterm prelabor rupture of membranes.

    Science.gov (United States)

    Kacerovsky, Marian; Musilova, Ivana; Khatibi, Ali; Skogstrand, Kristin; Hougaard, David M; Tambor, Vojtech; Tosner, Jindrich; Jacobsson, Bo

    2012-10-01

    To analyse whether intraamniotic inflammation in response to bacteria is different below and above gestational age 32 weeks in pregnancies complicated by preterm prelabor rupture of membranes (PPROM). A prospective study was performed, and 115 women with singleton pregnancies complicated by PPROM at gestational ages between 24(0/7) and 36(6/7) weeks were included in the study. Transabdominal amniocenteses were performed. Amniotic fluid was analysed using polymerase chain reactions for genital mycoplasmas and cultured for aerobic and anaerobic bacteria. The concentrations of 26 proteins in the amniotic fluid were determined simultaneously using multiplex technology. Bacteria were found in the amniotic fluid of 43% (49/115) of the women. The women were stratified into two subgroups according to gestational age 32 weeks. The amniotic fluid levels of four (interleukin-6, interleukin-10, CC chemokine ligands 2, and 3) and one specific (CC chemokine ligands 2) proteins were higher in women with the presence of bacteria in the amniotic fluid below and above 32 gestational weeks, respectively. An intraamniotic inflammatory response to bacteria in pregnancies complicated by PPROM seems to be different below and above 32 weeks of gestation.

  16. Amniotic mesenchymal stem cells display neurovascular tropism and aid in the recovery of injured peripheral nerves.

    Science.gov (United States)

    Li, YongNan; Guo, Longzhe; Ahn, Hyun Sook; Kim, Moo Hyun; Kim, Sung-Whan

    2014-06-01

    Recently, we reported that human amniotic membrane-derived mesenchymal stem cells (AMMs) possess great angiogenic potential. In this study, we determined whether local injection of AMMs ameliorates peripheral neuropathy. AMMs were transplanted into injured sciatic nerves. AMM injection promoted significant recovery of motor nerve conduction velocity and voltage amplitude compared to human adipose-derived mesenchymal stem cells. AMM implantation also augmented blood perfusion and increased intraneural vascularity. Whole-mount fluorescent imaging analysis demonstrated that AMMs exhibited higher engraftment and endothelial incorporation abilities in the sciatic nerve. In addition, the higher expression of pro-angiogenic factors was detected in AMMs injected into the peripheral nerve. Therefore, these data provide novel therapeutic and mechanistic insights into stem cell biology, and AMM transplantation may represent an alternative therapeutic option for treating peripheral neuropathy. © 2014 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

  17. Proteomic Analysis of Early Mid-Trimester Amniotic Fluid Does Not Predict Spontaneous Preterm Delivery

    Science.gov (United States)

    Lenco, Juraj; Vajrychova, Marie; Link, Marek; Tambor, Vojtech; Liman, Victor; Bullarbo, Maria; Nilsson, Staffan; Tsiartas, Panagiotis; Cobo, Teresa; Kacerovsky, Marian; Jacobsson, Bo

    2016-01-01

    Objective The aim of this study was to identify early proteomic biomarkers of spontaneous preterm delivery (PTD) in mid-trimester amniotic fluid from asymptomatic women. Methods This is a case-cohort study. Amniotic fluid from mid-trimester genetic amniocentesis (14–19 weeks of gestation) was collected from 2008 to 2011. The analysis was conducted in 24 healthy women with subsequent spontaneous PTD (cases) and 40 randomly selected healthy women delivering at term (controls). An exploratory phase with proteomics analysis of pooled samples was followed by a verification phase with ELISA of individual case and control samples. Results The median (interquartile range (IQR: 25th; 75th percentiles) gestational age at delivery was 35+5 (33+6–36+6) weeks in women with spontaneous PTD and 40+0 (39+1–40+5) weeks in women who delivered at term. In the exploratory phase, the most pronounced differences were found in C-reactive protein (CRP) levels, that were approximately two-fold higher in the pooled case samples than in the pooled control samples. However, we could not verify these differences with ELISA. The median (25th; 75th IQR) CRP level was 95.2 ng/mL (64.3; 163.5) in women with spontaneous PTD and 86.0 ng/mL (51.2; 145.8) in women delivering at term (p = 0.37; t-test). Conclusions Proteomic analysis with mass spectrometry of mid-trimester amniotic fluid suggests CRP as a potential marker of spontaneous preterm delivery, but this prognostic potential was not verified with ELISA. PMID:27214132

  18. Maternal plasma and amniotic fluid sphingolipids profiling in fetal Down syndrome.

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    Karol Charkiewicz

    Full Text Available Sphingolipids can be potentially involved in the formation of the central and peripheral nervous systems, which are particularly connected with the pathogenesis of Down syndrome. The aim of the study was to determine the concentration of selected sphingolipids in the plasma and amniotic fluid of pregnant patients with fetal Down syndrome.Out of 190 amniocentesis we had 10 patients with confirmed Down syndrome. For the purpose of our control we chose 14 women without confirmed chromosomal aberration. To assess the concentration of 11 sphingolipids in the blood plasma and amniotic fluid we used an ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC/MS/MS.We showed a significant increase in the concentration of 2 ceramides, C22-Cer and C24:1-Cer, in the plasma of women with fetal Down syndrome. Furthermore we showed a decrease in the concentration of 7 ceramides--C16-Cer, C18-Cer, C18:1-Cer, C20-Cer, C22-Cer, C24:1-Cer, and C24-Cer--in the amniotic fluid of women with fetal Down syndrome. We created ROC curves for all significant sphingolipids in maternal plasma, which set the threshold values and allowed for predicting the likelihood of Down syndrome in the fetus with specific sensitivity and specificity. We demonstrated a significantly higher risk of Down syndrome when the plasma concentration of C22-Cer > 12.66 ng/100 ul (sens. 0.9, sp. 0.79, P value = 0.0007 and C24:1-Cer > 33,19 ng/100 ul (sens. 0.6, sp. 0.86, P value = 0.0194.On the basis of our findings, it seems that the sphingolipids may play a role in the pathogenesis of Down syndrome. Defining their potential as biochemical markers of Down syndrome requires further investigation on a larger group of patients.

  19. Metabolomics of Human Amniotic Fluid and Maternal Plasma during Normal Pregnancy.

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    Magdalena Orczyk-Pawilowicz

    Full Text Available Metabolic profiles of amniotic fluid and maternal blood are sources of valuable information about fetus development and can be potentially useful in diagnosis of pregnancy disorders. In this study, we applied 1H NMR-based metabolic profiling to track metabolic changes occurring in amniotic fluid (AF and plasma (PL of healthy mothers over the course of pregnancy. AF and PL samples were collected in the 2nd (T2 and 3rd (T3 trimester, prolonged pregnancy (PP until time of delivery (TD. A multivariate data analysis of both biofluids reviled a metabolic switch-like transition between 2nd and 3rd trimester, which was followed by metabolic stabilization throughout the rest of pregnancy probably reflecting the stabilization of fetal maturation and development. The differences were further tested using univariate statistics at α = 0.001. In plasma the progression from T2 to T3 was related to increasing levels of glycerol, choline and ketone bodies (3-hydroxybutyrate and acetoacetate while pyruvate concentration was significantly decreased. In amniotic fluid, T2 to T3 transition was associated with decreasing levels of glucose, carnitine, amino acids (valine, leucine, isoleucine, alanine, methionine, tyrosine, and phenylalanine and increasing levels of creatinine, succinate, pyruvate, choline, N,N-dimethylglycine and urocanate. Lactate to pyruvate ratio was decreased in AF and conversely increased in PL. The results of our study, show that metabolomics profiling can be used to better understand physiological changes of the complex interdependencies of the mother, the placenta and the fetus during pregnancy. In the future, these results might be a useful reference point for analysis of complicated pregnancies.

  20. Transcriptome adaptation of group B Streptococcus to growth in human amniotic fluid.

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    Izabela Sitkiewicz

    Full Text Available BACKGROUND: Streptococcus agalactiae (group B Streptococcus is a bacterial pathogen that causes severe intrauterine infections leading to fetal morbidity and mortality. The pathogenesis of GBS infection in this environment is poorly understood, in part because we lack a detailed understanding of the adaptation of this pathogen to growth in amniotic fluid. To address this knowledge deficit, we characterized the transcriptome of GBS grown in human amniotic fluid (AF and compared it with the transcriptome in rich laboratory medium. METHODS: GBS was grown in Todd Hewitt-yeast extract medium and human AF. Bacteria were collected at mid-logarithmic, late-logarithmic and stationary growth phase. We performed global expression microarray analysis using a custom-made Affymetrix GeneChip. The normalized hybridization values derived from three biological replicates at each growth point were obtained. AF/THY transcript ratios representing greater than a 2-fold change and P-value exceeding 0.05 were considered to be statistically significant. PRINCIPAL FINDINGS: We have discovered that GBS significantly remodels its transcriptome in response to exposure to human amniotic fluid. GBS grew rapidly in human AF and did not exhibit a global stress response. The majority of changes in GBS transcripts in AF compared to THY medium were related to genes mediating metabolism of amino acids, carbohydrates, and nucleotides. The majority of the observed changes in transcripts affects genes involved in basic bacterial metabolism and is connected to AF composition and nutritional requirements of the bacterium. Importantly, the response to growth in human AF included significant changes in transcripts of multiple virulence genes such as adhesins, capsule, and hemolysin and IL-8 proteinase what might have consequences for the outcome of host-pathogen interactions. CONCLUSIONS/SIGNIFICANCE: Our work provides extensive new information about how the transcriptome of GBS responds

  1. Proteomic Analysis of Early Mid-Trimester Amniotic Fluid Does Not Predict Spontaneous Preterm Delivery.

    Science.gov (United States)

    Hallingström, Maria; Lenco, Juraj; Vajrychova, Marie; Link, Marek; Tambor, Vojtech; Liman, Victor; Bullarbo, Maria; Nilsson, Staffan; Tsiartas, Panagiotis; Cobo, Teresa; Kacerovsky, Marian; Jacobsson, Bo

    2016-01-01

    The aim of this study was to identify early proteomic biomarkers of spontaneous preterm delivery (PTD) in mid-trimester amniotic fluid from asymptomatic women. This is a case-cohort study. Amniotic fluid from mid-trimester genetic amniocentesis (14-19 weeks of gestation) was collected from 2008 to 2011. The analysis was conducted in 24 healthy women with subsequent spontaneous PTD (cases) and 40 randomly selected healthy women delivering at term (controls). An exploratory phase with proteomics analysis of pooled samples was followed by a verification phase with ELISA of individual case and control samples. The median (interquartile range (IQR: 25th; 75th percentiles) gestational age at delivery was 35+5 (33+6-36+6) weeks in women with spontaneous PTD and 40+0 (39+1-40+5) weeks in women who delivered at term. In the exploratory phase, the most pronounced differences were found in C-reactive protein (CRP) levels, that were approximately two-fold higher in the pooled case samples than in the pooled control samples. However, we could not verify these differences with ELISA. The median (25th; 75th IQR) CRP level was 95.2 ng/mL (64.3; 163.5) in women with spontaneous PTD and 86.0 ng/mL (51.2; 145.8) in women delivering at term (p = 0.37; t-test). Proteomic analysis with mass spectrometry of mid-trimester amniotic fluid suggests CRP as a potential marker of spontaneous preterm delivery, but this prognostic potential was not verified with ELISA.

  2. Proteomic Analysis of Early Mid-Trimester Amniotic Fluid Does Not Predict Spontaneous Preterm Delivery.

    Directory of Open Access Journals (Sweden)

    Maria Hallingström

    Full Text Available The aim of this study was to identify early proteomic biomarkers of spontaneous preterm delivery (PTD in mid-trimester amniotic fluid from asymptomatic women.This is a case-cohort study. Amniotic fluid from mid-trimester genetic amniocentesis (14-19 weeks of gestation was collected from 2008 to 2011. The analysis was conducted in 24 healthy women with subsequent spontaneous PTD (cases and 40 randomly selected healthy women delivering at term (controls. An exploratory phase with proteomics analysis of pooled samples was followed by a verification phase with ELISA of individual case and control samples.The median (interquartile range (IQR: 25th; 75th percentiles gestational age at delivery was 35+5 (33+6-36+6 weeks in women with spontaneous PTD and 40+0 (39+1-40+5 weeks in women who delivered at term. In the exploratory phase, the most pronounced differences were found in C-reactive protein (CRP levels, that were approximately two-fold higher in the pooled case samples than in the pooled control samples. However, we could not verify these differences with ELISA. The median (25th; 75th IQR CRP level was 95.2 ng/mL (64.3; 163.5 in women with spontaneous PTD and 86.0 ng/mL (51.2; 145.8 in women delivering at term (p = 0.37; t-test.Proteomic analysis with mass spectrometry of mid-trimester amniotic fluid suggests CRP as a potential marker of spontaneous preterm delivery, but this prognostic potential was not verified with ELISA.

  3. Isolation of a somatomedin binding protein from human preterm amniotic fluid: development of a radioimmunoassay

    International Nuclear Information System (INIS)

    Drop, S.L.S.

    1983-01-01

    This thesis investigates the nature and biological behaviour of a somatomedin binding protein, identified in preterm amniotic fluid (AF). For that purpose a double antibody radioimmunoassay was developed. Purified AF binding protein (AFBP) was iodinated by the chloramine-T method, and dilutions of partially purified AFBP were designated as the standard, with the results expressed in μg equivalent protein/ml. The sensitivity of the assay was improved by adoption of the nonequilibrium procedure. AFBP values were twice as high in preterm AF as in term AF. (Auth.)

  4. Consequences of meconium stained amniotic fluid: what does the evidence tell us?

    Science.gov (United States)

    Hutton, Eileen K; Thorpe, Julia

    2014-07-01

    Meconium stained amniotic fluid (MSAF) is common and associated with meconium aspiration syndrome (MAS). Other consequences of meconium passage before birth are less well understood. We reviewed the literature for original papers reporting on outcomes associated with MSAF. Among preterm infants MSAF is more prevalent than previously believed and is associated with higher neonatal morbidity. Intrauterine exposure to meconium is associated with inflammation of tissues of the lung, chorionic plate and umbilical vessels and through various mechanisms may contribute to neonatal morbidity, independent of MAS. No compelling evidence supported an association between MSAF and increased neurological impairment, including early seizure activity. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Does amniotic fluid volume affect fetofetal transfusion in monochorionic twin pregnancies? Modelling two possible mechanisms

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    Umur, Asli; van Gemert, Martin J. C.; Ross, Michael G.

    2002-06-01

    Clinical evidence suggests that increased amniotic fluid volume due to polyhydramnios increases placental vascular resistance. We have sought to model the possible effects of an increased amniotic fluid volume on the net fetofetal transfusion in monochorionic twin pregnancies. We wanted to compare these effects with the results of previous simulations, which aimed to explain why the twin-twin transfusion syndrome (TTTS) placentas more often include bidirectional arteriovenous (AV) rather than AV plus arterioarterial (AA) anastomoses. We extended our mathematical model of TTTS by simulating two different mechanisms that increase the placental vascular resistance as a consequence of polyhydramnios. First, there is an increase in the placental capillary resistance and hence in deep AV and opposite AV (denoted as VA) resistances due to polyhydramnios. Second, there is an increase in the resistance of chorionic veins due to polyhydramnios, assuming that these veins act as Starling resistors. We then simulated the effects of polyhydramnios on different placental anastomotic patterns. The results were as follows. In the first mechanism (polyhydramnios affects AV-VA resistances), an increased amniotic fluid volume hardly affected bidirectional AV, but slightly decreased fetofetal transfusion in AV plus AA anastomoses. However, for these effects to change the natural development of the pregnancy, polyhydramnios needed to persist for approximately 4 weeks, and by comparing the effects of polyhydramnios with the effects of amnioreduction, amnioreduction was more beneficial for normalizing the donor amniotic fluid volume. Therefore, these beneficial effects due to polyhydramnios have no practical clinical significance. In the second mechanism (Starling resistor for chorionic veins), polyhydramnios slightly increased fetofetal transfusion and hence slightly increased TTTS severity in bidirectional AV and AV plus VV, but did not affect AV plus AA anastomoses. In conclusion, we

  6. The Effects of Intravenous Hydration on Amniotic Fluid Volume and Pregnancy Outcomes in Women with Term Pregnancy and Oligohydramnios: A Randomized Clinical Trial

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    Mahnaz Shahnazi

    2012-08-01

    Full Text Available Introduction: Amniotic fluid is an important factor in the prediction of fetal survival. The aim of this research was to evaluate the effects of intravenous hydration of mothers on amniotic fluid volume and in turn on pregnancy outcomes. Methods: The current single blind controlled clinical trial was conducted on 20 pregnant mothers with amniot-ic fluid index of lower or equal to 5 cm and gestational age of 37-41 weeks. The subjects were divided into two groups of case and control through simple random sampling. Am-niotic fluid index was measured in all participants. The case group received one liter of isotonic saline during 30 minutes by the bolus method. Reevaluations of amniotic fluid index in both groups were made 90 minutes after baseline measurement. Independent t-test and paired t-test were used to compare the two groups and mean amniotic fluid in-dex before and after treatment, respectively. Results: Hydration of mothers significantly increased the amniotic fluid index in the case group (mean change: 1.5 cm; 95%CI: 0.46 - 2.64; P = 0.01. The mean change of amniotic fluid index in the control group did not significantly increase (P = 0.06. The elevation of amniotic fluid index in the hydra-tion group (32% was significantly higher than the control group (1% (P = 0.03. Conclusion: In this study intravenous hydration increased amniotic fluid index of mothers with term pregnancy and oligohydramnios. Since it caused no complications for the moth-er and the fetus, it can be used as an effective method in management of oligohydramnios.

  7. Second trimester amniotic fluid myo-inositol concentrations in women later developing gestational diabetes mellitus or pregnancy-induced hypertension.

    Science.gov (United States)

    Santamaria, Angelo; Corrado, Francesco; Baviera, Giovanni; Carlomagno, Gianfranco; Unfer, Vittorio; D'anna, Rosario

    2016-01-01

    To evaluate myo-inositol concentrations in amniotic fluid in women later developing gestational diabetes and hypertension. A retrospective study was carried out with three groups of amniotic fluid samples (15-18 gestational weeks): 30 gestational hypertension pregnancies, 30 gestational diabetes pregnancies, and 30 normal pregnancy. A significant difference was observed in myo-inositol concentrations between the median gestational diabetes values (124.0 µmol/L, IQR 90.0-162.5) and the control group values (79.0 µmol/L, IQR 62.0-107.5), but also with gestational hypertension median values (79.0 µmol/L, IQR 67.75-92.0) (p inositol concentrations in amniotic fluid increased significantly in women later developing gestational diabetes compared to the control group.

  8. Human amniotic fluid contaminants alter thyroid hormone signalling and early brain development in Xenopus embryos

    Science.gov (United States)

    Fini, Jean-Baptiste; Mughal, Bilal B.; Le Mével, Sébastien; Leemans, Michelle; Lettmann, Mélodie; Spirhanzlova, Petra; Affaticati, Pierre; Jenett, Arnim; Demeneix, Barbara A.

    2017-03-01

    Thyroid hormones are essential for normal brain development in vertebrates. In humans, abnormal maternal thyroid hormone levels during early pregnancy are associated with decreased offspring IQ and modified brain structure. As numerous environmental chemicals disrupt thyroid hormone signalling, we questioned whether exposure to ubiquitous chemicals affects thyroid hormone responses during early neurogenesis. We established a mixture of 15 common chemicals at concentrations reported in human amniotic fluid. An in vivo larval reporter (GFP) assay served to determine integrated thyroid hormone transcriptional responses. Dose-dependent effects of short-term (72 h) exposure to single chemicals and the mixture were found. qPCR on dissected brains showed significant changes in thyroid hormone-related genes including receptors, deiodinases and neural differentiation markers. Further, exposure to mixture also modified neural proliferation as well as neuron and oligodendrocyte size. Finally, exposed tadpoles showed behavioural responses with dose-dependent reductions in mobility. In conclusion, exposure to a mixture of ubiquitous chemicals at concentrations found in human amniotic fluid affect thyroid hormone-dependent transcription, gene expression, brain development and behaviour in early embryogenesis. As thyroid hormone signalling is strongly conserved across vertebrates the results suggest that ubiquitous chemical mixtures could be exerting adverse effects on foetal human brain development.

  9. Amniotic fluid for ex vivo skin preservation: a comparative study of tissue preservation solutions.

    Science.gov (United States)

    Buseman, Jason; Rinker, Alexander B; Rinker, Brian

    2013-12-01

    Ex vivo skin preservation is important for skin banks, burn centers, and in research; however, the optimal preservation solution is not known. Human amniotic fluid (HAF), in addition to its role in fetal wound healing, has promise as an effective and readily available preservation solution. The purpose of this study was to compare the efficacy of several solutions, including HAF, in full-thickness skin preservation. Human amniotic fluid was obtained from patients undergoing amniocentesis. Full-thickness skin obtained during abdominoplasty was divided into 1-cm(2) samples. These specimens were preserved in either saline, HAF from a single patient, pooled HAF, University of Wisconsin solution, or custodial histidine-tryptophan-ketoglutarate solution at 4°C. There were 5 samples in each group. Specimens were examined for keratinocyte survival at 7, 14, 21, 28, and 35 days using the trypan blue assay. The first 200 cells identified were counted to calculate the degree of cell death. Comparisons were made between the groups, and a multivariable repeated-measures analysis was performed to determine statistical significance, which was defined as P skin banks, burn centers, and research.

  10. Prooxidant-Antioxidant Balance in Umbilical Cord Blood of Infants with Meconium Stained of Amniotic Fluid

    Directory of Open Access Journals (Sweden)

    Mohammad Hassan Arjmand

    2013-01-01

    Full Text Available Objective. Using a novel assay termed prooxidant-antioxidant balance (PAB assay to determine prooxidant-antioxidant balance in umbilical cord blood of infants with meconium stained of amniotic fluid (MSAF. Passage of meconium in amniotic fluid is associated with increase of neonatal mortality and morbidity. This complication occurs in about 15% of infants and is more widespread in postterm neonates. About 1.5 percent of neonates with MSAF develop meconium aspiration syndrome. Method. Sera of 29 umbilical cord blood of infants with MSAF and 32 healthy infants (HI were collected. Both groups had nonsmoker and non-alcoholic mothers with no diseases. The PAB was measured. Result. There was a significant increase of PAB value (32.8±15.9 HK in umbilical cord blood of infants with MSAF in comparison to HI (24.5±12.6 HK (P<0.05. There was no significant correlation between PAB value and age of mothers. Conclusion. The increased PAB value in infants with MSAF showed that these infants are exposed to oxidative stress. Further research with larger population is needed to demonstrate the oxidative stress in infants with MSAF.

  11. Associations of viscosity, stercobilin and bilirubin levels in meconium stained amniotic fluid to meconium aspiration syndrome

    Directory of Open Access Journals (Sweden)

    M. Sholeh Kosim

    2011-04-01

    Full Text Available Background Meconium-stained amniotic fluid (MSAF increases morbidity and mortality in neonates. Meconium aspiration syndrome (MAS occurs in 2-9% of neonates with MSAF. Viscosity of MSAF is associated with the amount of the meconium release. Objective To determine the associations between viscosity and the presence of stercobilin and bilirubin in MSAF with MAS in neonates. Methods This observational cohort study was perfonned with term babies who were born v.ith MSAF in Kariadi Hospital from August 2009 to May 2010. Amniotic fluid specimens were taken at birth and neonates were observed for respiratory symptoms until the 5th day of life. Analysis was done by chi-square test, Fisher's exact test and relative risk. Results The majority of the 48 subjects were male, Mth mean gestational age of 39.9 (SD 1.73 weeks. Classification of MSAF as thick or thin was done by macroscopic examination Mth Kappa test 0.741. The MSAF tested ositively for stercobilin and bilirubin in 12/48 and 17/48 subjects, respectively. Thick MSAF correlated significantly to MAS (P=0.03 Mth a relative risk of 10.1 (95% CI 1.2 to 87 .6, while stercobilin and bilirubin presence did not. Conclusion Thick MSAF was associated Mth lvtAS and was a risk factor for MAS. Stercobilin and bilirubin presence in MSAF were not associated with MAS.

  12. The association between mean platelet volume and infants with meconium stained amniotic fluid.

    Science.gov (United States)

    Tekin, M; Konca, C; Kahramaner, Z; Erdemir, A; Gulyuz, A; Uckardes, F; Turgut, M

    2016-07-01

    The exact pathophysiology of meconium passage into the amniotic fluid is unknown, but it is frequently associated with fetal hypoxia. The mean platelet volume (MPV) seems to be a marker of platelet production and consumption and may be related to the severity of some diseases associated with bone marrow, hypoxia, and perinatal infections. We aimed to investigate the association between MPV levels and meconium-stained amniotic fluid (MSAF) in infants. MPV, serum-reactive protein and hemoglobin levels, and leukocyte and thrombocyte counts were measured in 106 infants with MSAF and a comparison group of 78 healthy control infants. The mean MPV values of the infants with MSAF were statistically significantly lower than those of the control group (p 0.05). There was also no statistically significant difference in the MPV levels of the infants with meconium aspiration syndrome (MAS) compared to the infants with MSAF without MAS (p = 0.107). The optimal cut-off value for the MPV was 9.90 fl (area under the curve [AUC: 0.788]) in the infants with MSAF, with a sensitivity of 78.1% and specificity of 74.3%. Our data suggest that the MPV levels of infants with MSAF were significantly lower than those of healthy infants. This might be associated with a hypoxic process. However, the MPV levels of infants with MSAF and MAS were statistically similar. Thus, the MPV level could not be used to detect patients with or without severe disease.

  13. Amniotic fluid

    Science.gov (United States)

    Burton GJ, Sibley CP, Jauniaux ERM. Placental anatomy and physiology. In: Gabbe SG, Niebyl JR, Simpson JL, et al, eds. Obstetrics: Normal and Problem Pregnancies . 7th ed. Philadelphia, PA: Elsevier; 2017:chap 1. ...

  14. The effect of electromagnetic radiation of wireless connections on morphology of amniotic fluid

    Science.gov (United States)

    Novikov, Vsevolod O.; Titova, Natalia; Azarhov, Olexand; Wójcik, Waldemar; GrÄ dz, Å.»aklin; Mussabekova, Assel

    2016-09-01

    The article considers the effect of wireless networks on the morphology of amniotic fluid (AF) to demonstrate possible risks involving pregnant women. The analysis of AF thesiograms after exposure of the model fluid to Wi-Fi, 3G and β- radiation was chosen as the research method. A comparative analysis of facies structures is carried out, and depth maps of the facies structure are created. This comparative analysis permits an evaluation of the efficiency of morphological changes. It is shown that AF control facies differ in the concentration of areas with a narrow peripheral area and ellipsoidal formations of crystalloids in circumferences center. After exposure of different types of radiation onto AF, the facies structures collapse and form their own conglomerates. The obtained results show that the considered types of radiation have a negative effect on AF.

  15. The presence of vaginal Lactobacillus species does not contribute to a measureable difference in amniotic fluid lactate levels collected from the vaginal tract of laboring women.

    Science.gov (United States)

    Hall, Beverley; Wong, Diana; Healy, Clare; Tracy, Mark B; Tracy, Sally K; Rawlinson, William D

    2017-04-01

    Amniotic fluid lactate research is based on the hypothesis that a relationship exists between fatigued uterine muscles and raised concentrations of the metabolite lactate, which is excreted into the amniotic fluid during labor. To assess potentially confounding effects of lactate-producing organisms on amniotic fluid lactate measurements, we aimed to determine if the presence of vaginal Lactobacillus species was associated with elevated levels of amniotic fluid lactate, measured from the vaginal tract of women in labor. Results from this study contribute to a large prospective longitudinal study of amniotic fluid lactate at a teaching hospital in Sydney, Australia. Amniotic fluid lactate measurement was assessed at the time of routine vaginal examination, after membranes had ruptured, using a hand-held lactate meter StatStripXPress (Nova Biomedical). Vaginal swab samples were collected at the time of the first amniotic fluid lactate measurement and stored for later detection and quantification of Lactobacillus species using a TaqMan real-time PCR assay. Swab sample and amniotic fluid lactate results were paired and analyzed. The PCR assay detected Lactobacillus species in 48 of 388 (12%) vaginal swab specimens (8% positive, 4% low positive) collected from women in labor after membranes had ruptured. There was no significant difference in median and mean (respectively) amniotic fluid lactate levels with (8.35 mmol/L; 8.95 mmol/L) or without (8.5 mmol/L; 9.08 mmol/L) Lactobacillus species detected. There was no association between the presence or level of vaginal Lactobacillus species and the measurement of amniotic fluid lactate collected from the vaginal tract of women during labor. © 2017 Nordic Federation of Societies of Obstetrics and Gynecology.

  16. Amniotic Fluid Soluble Myeloid Differentiation-2 (sMD-2) as Regulator of Intra-amniotic Inflammation in Infection-induced Preterm Birth.

    Science.gov (United States)

    Dulay, Antonette T; Buhimschi, Catalin S; Zhao, Guomao; Oliver, Emily A; Abdel-Razeq, Sonya S; Shook, Lydia L; Bahtiyar, Mert O; Buhimschi, Irina A

    2015-06-01

    TLR4 mediates host responses to pathogens through a mechanism that involves protein myeloid differentiation-2 (MD-2) and its soluble form sMD-2. The role of sMD2 in intra-amniotic inflammation-induced preterm birth has not been previously explored. Human amniotic fluid (AF) sMD-2 was studied by Western blotting in 152 AF samples of patients who had an amniocentesis to rule-out infection (yes infection, n = 50; no infection, n = 50) or women with normal pregnancy outcome (second trimester genetic karyotyping, n = 26; third trimester lung maturity testing, n = 26). Histological localization and mRNA expression of MD2 in fetal membranes were studied by immunohistochemistry and RT-PCR. The ability of fetal membrane to release sMD-2 and inflammatory cytokines was studied in vitro. Human AF contains three sMD-2 proteoforms whose levels of expression were lower at term. Intra-amniotic infection upregulated sMD-2. MD-2 mRNA and immunohistochemistry findings concurred. In vitro, LPS and monensin increased, while cycloheximide decreased sMD-2 production. Recombinant sMD-2 modulated TNF-α and IL-6 levels in a dose- and time-dependent fashion. sMD2 proteoforms are constitutively present in human AF. The intensity of the intra-amniotic inflammatory response to bacteria or perhaps to other TLR4 ligands may be facilitated through synthesis and release of sMD2 by the amniochorion. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. More than fetal urine: enteral uptake of amniotic fluid as a major predictor for fetal growth during late gestation

    NARCIS (Netherlands)

    Bagci, S.; Brosens, E.; Tibboel, D.; Klein, A.; Ijsselstijn, H.; Wijers, C.H.W.; Roeleveld, N.; Blaauw, I. de; Broens, P.M.; Rooij, I.A.L.M. van; Holscher, A.; Boemers, T.M.; Pauly, M.; Munsterer, O.J.; Schmiedeke, E.; Schafer, M.; Ure, B.E.; Lacher, M.; Choinitzki, V.; Schumacher, J.; Zwink, N.; Jenetzky, E.; Katzer, D.; Arand, J.; Bartmann, P.; Reutter, H.M.

    2016-01-01

    The purpose of our study was to investigate the importance of amniotic fluid (AF) for fetal growth during late gestation using esophageal atresia (EA) patients as a model. In this retrospective cohort study, we compared the z-scores adapted for birth weights (BW z-scores) for each of 517 European

  18. More than fetal urine : enteral uptake of amniotic fluid as a major predictor for fetal growth during late gestation

    NARCIS (Netherlands)

    Bagci, Soyhan; Brosens, Erwin; Tibboel, Dick; De Klein, Annelies; Ijsselstijn, Hanneke; Wijers, Charlotte H. W.; Roeleveld, Nel; de Blaauw, Ivo; Broens, Paul M.; van Rooij, Iris A. L. M.; Hoelscher, Alice; Boemers, Thomas M.; Pauly, Marcus; Muensterer, Oliver J.; Schmiedeke, Eberhard; Schaefer, Mattias; Ure, Benno E.; Lacher, Martin; Choinitzki, Vera; Schumacher, Johannes; Zwink, Nadine; Jenetzky, Ekkehart; Katzer, David; Arand, Joerg; Bartmann, Peter; Reutter, Heiko M.

    The purpose of our study was to investigate the importance of amniotic fluid (AF) for fetal growth during late gestation using esophageal atresia (EA) patients as a model. In this retrospective cohort study, we compared the z-scores adapted for birth weights (BW z-scores) for each of 517 European

  19. Characterisation of non-maternal serum proteins in amniotic fluid at weeks 16 to 18 of gestation

    DEFF Research Database (Denmark)

    Drøhse, H; Christensen, H; Myrhøj, Vibeke

    1998-01-01

    of these proteins were chosen for isolation and further characterisation. With the use of immunological methods, SDS-PAGE and N-terminal sequencing we identified two of the proteins as C-terminal propeptides of procollagen Type I and Type III, which have not hitherto been described in amniotic fluid. The third...

  20. In vivo (1)H magnetic resonance spectroscopy of amniotic fluid and fetal lung at 1.5 T: technical challenges.

    Science.gov (United States)

    Kim, Dong-Hyun; Vahidi, Kiarash; Caughey, Aaron B; Coakley, Fergus V; Vigneron, Daniel B; Kurhanewicz, John; Mow, Ben; Joe, Bonnie N

    2008-10-01

    To identify the major technical challenges associated with in utero single-voxel proton spectroscopy of amniotic fluid and fetal lung and to evaluate the feasibility of performing in utero fetal spectroscopy for fetal lung maturity testing. Fetal magnetic resonance (MR) spectroscopy of amniotic fluid and fetal lung were performed at 1.5 T in 8 near-term pregnant women. Presence/absence of lactate and choline peaks was tabulated. Ex vivo spectra were obtained from amniotic fluid samples to investigate and refine sequence parameters. Spectroscopy failed in 3 of 8 cases due to maternal discomfort (n = 1) or fetal gastroschisis (n = 2). Both fetal motion and low signal-to-noise ratio were limiting factors for the remaining 5 clinical in vivo studies at 1.5 T. Ex vivo and in vivo studies suggested feasibility for detecting lactate from amniotic fluid within a reasonable clinical scan time (4-5 minutes). Lactate was detected in 3 of 5 patients. Choline detection was limited and was detected in 1 patient. Minor motion effects can be overcome but continuous fetal motion is problematic. Lactate detection seems clinically feasible, but choline detection requires additional technical development and, potentially, further imaging at a higher field strength because of the low signal-to-noise ratio at 1.5 T. (c) 2008 Wiley-Liss, Inc.

  1. Chest computed tomography of a patient revealing severe hypoxia due to amniotic fluid embolism: a case report

    Directory of Open Access Journals (Sweden)

    Inui Daisuke

    2010-02-01

    Full Text Available Abstract Introduction Amniotic fluid embolism is one of the most severe complications in the peripartum period. Because its onset is abrupt and fulminant, it is unlikely that there will be time to examine the condition using thoracic computed tomography (CT. We report a case of life-threatening amniotic fluid embolism, where chest CT in the acute phase was obtained. Case presentation A 22-year-old Asian Japanese primiparous woman was suspected of having an amniotic fluid embolism. After a Cesarean section for cephalopelvic disproportion, her respiratory condition deteriorated. Her chest CT images were examined. CT findings revealed diffuse homogeneous ground-glass shadow in her bilateral peripheral lung fields. She was therefore transferred to our hospital. On admission to our hospital's intensive care unit, she was found to have severe hypoxemia, with SpO2 of 50% with a reservoir mask of 15 L/min oxygen. She was intubated with the support of noninvasive positive pressure ventilation. She was successfully extubated on the sixth day, and discharged from the hospital on the twentieth day. Conclusion This is the first case report describing amniotic fluid embolism in which CT revealed an acute respiratory distress syndrome-like shadow.

  2. Affinity purification of native glycodelin from amniotic fluid for biological investigations and development of a glycodelin ELISA for clinical studies

    DEFF Research Database (Denmark)

    Sørensen, Steen; Myrhøj, Vibeke; Nguyen, Thanh Ha

    2017-01-01

    for functional studies because the carbohydrate part can be lacking or be insufficient in recombinant glycodelin from prokaryotic and eukaryotic cell systems. METHODS AND RESULTS: Native glycodelin was purified from amniotic fluid by a series of affinity chromatography steps and had many glycosylated forms...

  3. Analysis of perchlorate, thiocyanate, nitrate and iodide in human amniotic fluid using ion chromatography and electrospray tandem mass spectrometry

    International Nuclear Information System (INIS)

    Blount, Benjamin C.; Valentin-Blasini, Liza

    2006-01-01

    Because of health concerns surrounding in utero exposure to perchlorate, we developed a sensitive and selective method for quantifying iodide, as well as perchlorate and other sodium-iodide symporter (NIS) inhibitors in human amniotic fluid using ion chromatography coupled with electrospray ionization tandem mass spectrometry. Iodide and NIS inhibitors were quantified using a stable isotope-labeled internal standards (Cl 18 O 4 - , S 13 CN - and 15 NO 3 - with excellent assay accuracy of 100%, 98%, 99%, 95% for perchlorate, thiocyanate, nitrate and iodide, respectively, in triplicate analysis of spiked amniotic fluid sample). Excellent analytical precision (<5.2% RSD for all analytes) was found when amniotic fluid quality control pools were repetitively analyzed for iodide and NIS-inhibitors. Selective chromatography and tandem mass spectrometry reduced the need for sample cleanup, resulting in a rugged and rapid method capable of routinely analyzing 75 samples/day. Analytical response was linear across the physiologically relevant concentration range for the analytes. Analysis of a set of 48 amniotic fluid samples identified the range and median levels for perchlorate (0.057-0.71, 0.18 μg/L), thiocyanate (<10-5860, 89 μg/L), nitrate (650-8900, 1620 μg/L) and iodide (1.7-170, 8.1 μg/L). This selective, sensitive, and rapid method will help assess exposure of the developing fetus to low levels of NIS-inhibitors and their potential to inhibit thyroid function

  4. Prenatal cytogenetic diagnosis in Spain: analysis and evaluation of the results obtained from amniotic fluid samples during the last decade.

    Science.gov (United States)

    Mademont-Soler, Irene; Morales, Carme; Clusellas, Núria; Soler, Anna; Sánchez, Aurora

    2011-08-01

    Chromosome abnormalities are one of the main causes of congenital defects, and establishing their frequency according to the different clinical indications for invasive procedure during pregnancy is especially important for genetic counselling. We analyzed the results of 29,883 amniotic fluid samples referred to our laboratory for cytogenetic studies from 1998 to 2009, which constitutes the largest series of cytogenetic analysis performed on amniotic fluid samples in Spain. The number of samples received tended to increase from 1998 to 2005, but after 2005 it decreased substantially. Cytogenetic results were obtained in 99.5% of the samples, and the detected incidence of chromosome abnormalities was 2.9%. Of these, 48.1% consisted of classical autosomal aneuploidies, trisomy 21 being the most frequent one. The main clinical indications for amniocentesis were positive prenatal screening and advanced maternal age, but referral reasons with highest positive predictive values were, excluding parental chromosome rearrangement, increased nuchal translucency (9.2%) and ultrasound abnormalities (6.6%). In conclusion, performing the karyotype on amniotic fluid samples is a good method for the detection of chromosome abnormalities during pregnancy. The number of cytogenetic studies on amniotic fluid has now decreased, however, due to the implementation of first trimester prenatal screening for the detection of Down syndrome, which allows karyotyping on chorionic villus samples. Our results also show that both ultrasound abnormalities and increased nuchal translucency are excellent clinical indicators for fetal chromosome abnormality. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  5. Amniotic membrane-derived stem cells: immunomodulatory properties and potential clinical application

    Directory of Open Access Journals (Sweden)

    Insausti CL

    2014-03-01

    Full Text Available Carmen L Insausti,1 Miguel Blanquer,1 Ana M García-Hernández,1 Gregorio Castellanos,2 José M Moraleda11Unidad de Trasplante Hematopoyético y Terapia Celular, 2Servicio de Cirugía, Hospital Clínico Universitario Virgen de la Arrixaca, IMIB, Campus Mare Nostrum, Universidad de Murcia, El Palmar, Murcia, SpainAbstract: Epithelial and mesenchymal cells isolated from the amniotic membrane (AM possess stem cell characteristics, differentiation potential toward lineages of different germ layers, and immunomodulatory properties. While their expansion and differentiation potential have been well studied and characterized, knowledge about their immunomodulatory properties and the mechanisms involved is still incomplete. These mechanisms have been evaluated on various target cells of the innate and the adaptive system and in animal models of different inflammatory diseases. Some results have evidenced that the immunomodulatory effect of AM-derived cells is dependent on cell-cell contact, but many of them have demonstrated that these properties are mediated through the secretion of suppressive molecules. In this review, we present an update on the described immunomodulatory properties of the derived amniotic cells and some of the proposed involved mechanisms. Furthermore, we describe some assays in animal models of different inflammatory diseases which reveal the potential use of these cells to treat such diseases.Keywords: epithelial cells, mesenchymal cells, cell therapy, immunomodulation

  6. ROLE OF AMNIOINFUSION ON NEONATAL OUTCOME IN CASES WITH MECONIUMSTAINED AMNIOTIC FLUID

    Directory of Open Access Journals (Sweden)

    K. Chandramathy

    2016-10-01

    Full Text Available BACKGROUND Amnioinfusion is thought to dilute meconium present in the amniotic fluid and so reduce the risk of meconium aspiration in newborn. AIM The effect of amnioinfusion in meconium-stained amniotic fluid in reducing the incidence of meconium aspiration syndrome, hypoxic ischaemic encephalopathy and perinatal mortality in newborn. MATERIALS AND METHODS The study was carried out in the Departments of Obstetrics and Gynaecology and NICU at Government Medical College, Kozhikode, in a time duration of one year from January 2014 to December 2014. This is a prospective case control study. We have studied 210 antenatal women admitted to the labour room with grade 2/3 meconium-stained amniotic fluid after 36 weeks of gestation. Amnioinfusion was given in 140 cases and 70 cases given standard care. Patients were monitored with electronic foetal heart monitoring and caesarean section was done in case of foetal distress or those who are in early labour. There was no significant difference between study group and control group according to age, parity, gestational age, presence of complications like hypertension/pre-eclampsia, post-dated pregnancy, anaemia, etc. RESULTS Foetal heart rate decelerations occurred in 27 out of 140 cases (19% in the study group and 23 out of 70 (33% in control group (P <0.05. NEONATAL OUTCOME In our study those who received amnioinfusion, only 19% delivered babies with APGAR <9 at 1’ while those who do not received, 36% delivered babies with APGAR <9 at 1’ (P value of 0.009. Meconium aspiration syndrome occurred in 2.1% of cases in the infusion group and 11.4% in the non-infusion group (P<0.005. Respiratory distress was markedly reduced in amnioinfusion group 28% compared to 63% in controls (P=0.002. NICU admissions were 64% in control group compared to 22% in amnioinfusion group. Perinatal mortality and hypoxic ischaemic encephalopathy in the infusion group were nil as compared to 7% and 11% in the control group

  7. Phthalates and perfluorooctanesulfonic acid in human amniotic fluid: temporal trends and timing of amniocentesis in pregnancy.

    Science.gov (United States)

    Jensen, Morten Søndergaard; Nørgaard-Pedersen, Bent; Toft, Gunnar; Hougaard, David M; Bonde, Jens Peter; Cohen, Arieh; Thulstrup, Ane Marie; Ivell, Richard; Anand-Ivell, Ravinder; Lindh, Christian H; Jönsson, Bo A G

    2012-06-01

    Measures of prenatal environmental exposures are important, and amniotic fluid levels may directly reflect fetal exposures during hypothesized windows of vulnerability. We aimed to detect various phthalate metabolites and perfluorooctanesulfonic acid (PFOS) in human amniotic fluid, to study temporal exposure trends, and to estimate potential associations with gestational week of amniocentesis and maternal age and parity at amniocentesis. We studied 300 randomly selected second-trimester amniotic fluid samples from a Danish pregnancy-screening biobank covering 1980 through 1996. We used only samples from male offspring pregnancies. We assayed the environmental pollutants by liquid chromatography/triple quadrupole mass spectrometry and analyzed data using generalized linear regression models. We detected the di(2-ethylhexyl) phthalate (DEHP) metabolite mono(2-ethyl-5-carboxypentyl) phthalate (5cx-MEPP) at a median concentration of 0.27 ng/mL [interquartile range (IQR): 0.20-0.37 ng/mL], the diisononyl phthalate (DiNP) metabolite mono(4-methyl-7-carboxyheptyl) phthalate (7cx-MMeHP) at 0.07 ng/mL (IQR: 0.05-0.11 ng/mL), and PFOS at 1.1 ng/mL (IQR: 0.66-1.60 ng/mL). An increase of 1 calendar year was associated with 3.5% lower [95% confidence interval (CI): -4.8%, -2.1%] 5cx-MEPP levels and with 7.1% higher (95% CI: 5.3%, 9.0%) 7cx-MMeHP levels. For each later gestational week of amniocentesis, 5cx-MEPP was 9.9% higher (95% CI: 4.8%, 15.2%), 7cx-MMeHP was 8.6% higher (95: CI: 2.7%, 14.9%), and PFOS was 9.4% higher (95: CI: 3.3%, 15.9%). We observed no associations with maternal age or parity. Measured metabolite levels appeared to parallel decreasing DEHP exposure and increasing DiNP exposure during the study period. The environmental pollutant levels were positively associated with later gestational age at amniocentesis during pregnancy weeks 12-22.

  8. Reconstruction of limbal stem cell deficient corneal surface with induced human bone marrow mesenchymal stem cells on amniotic membrane.

    Science.gov (United States)

    Rohaina, Che Man; Then, Kong Yong; Ng, Angela Min Hwei; Wan Abdul Halim, Wan Haslina; Zahidin, Aida Zairani Mohd; Saim, Aminuddin; Idrus, Ruszymah B H

    2014-03-01

    The cornea can be damaged by a variety of clinical disorders or chemical, mechanical, and thermal injuries. The objectives of this study were to induce bone marrow mesenchymal stem cells (BMSCs) to corneal lineage, to form a tissue engineered corneal substitute (TEC) using BMSCs, and to treat corneal surface defects in a limbal stem cell deficiency model. BMSCs were induced to corneal lineage using limbal medium for 10 days. Induced BMSCs demonstrated upregulation of corneal stem cell markers; β1-integrin, C/EBPδ, ABCG2, and p63, increased protein expression of CK3 and p63 significantly compared with the uninduced ones. For TEC formation, passage 1 BMSCs were trypsinized and seeded on amniotic membrane in a transwell co-culture system and were grown in limbal medium. Limbal stem cell deficiency models were induced by alkaline injury, and the TEC was implanted for 8 weeks. Serial slit lamp evaluation revealed remarkable improvement in corneal regeneration in terms of corneal clarity and reduced vascularization. Histologic and optical coherence tomography analyses demonstrated comparable corneal thickness and achieved stratified epithelium with a compact stromal layer resembling that of normal cornea. CK3 and p63 were expressed in the newly regenerated cornea. In conclusion, BMSCs can be induced into corneal epithelial lineage, and these cells are viable for the formation of TEC, to be used for the reconstruction of the corneal surface in the limbal stem cell deficient model. Copyright © 2014 Mosby, Inc. All rights reserved.

  9. Biocompatibility of quantum dots (CdSe/ZnS ) in human amniotic membrane-derived mesenchymal stem cells in vitro.

    Science.gov (United States)

    Wang, Gongping; Zeng, Guangwei; Wang, Caie; Wang, Huasheng; Yang, Bo; Guan, Fangxia; Li, Dongpeng; Feng, Xiaoshan

    2015-06-01

    Amniotic membrane-derived mesenchymal stem cells (hAM-dMSCs) are a potential source of mesenchymal stem cells which could be used to repair skin damage. The use of mesenchymal stem cells to repair skin damage requires safe, effective and biocompatible agents to evaluate the effectiveness of the result. Quantum dots (QDs) composed of CdSe/ZnS are semiconductor nanocrystals with broad excitation and narrow emission spectra, which have been considered as a new chemical and fluorescent substance for non-invasively labeling different cells in vitro and in vivo. This study investigated the cytotoxic effects of QDs on hAM-dMSCs at different times following labeling. Using 0.75, 1.5 and 3.0 μL between quantum dots, labeled human amniotic mesenchymal stem cells were collected on days 1, 2 and 4 and observed morphological changes, performed an MTT cell growth assay and flow cytometry for mesenchymal stem cells molecular markers. Quantum dot concentration 0.75 μg/mL labeled under a fluorescence microscope, cell morphology was observed, The MTT assay showed cells in the proliferative phase. Flow cytometry expression CD29, CD31, CD34, CD44, CD90, CD105 and CD106. Within a certain range of concentrations between quantum dots labeled human amniotic mesenchymal stem cells has good biocompatibility.

  10. Amniotic fluid deficiency and congenital abnormalities both influence fluctuating asymmetry in developing limbs of human deceased fetuses.

    Directory of Open Access Journals (Sweden)

    Clara Mariquita Antoinette ten Broek

    Full Text Available Fluctuating asymmetry (FA, as an indirect measure of developmental instability (DI, has been intensively studied for associations with stress and fitness. Patterns, however, appear heterogeneous and the underlying causes remain largely unknown. One aspect that has received relatively little attention in the literature is the consequence of direct mechanical effects on asymmetries. The crucial prerequisite for FA to reflect DI is that environmental conditions on both sides should be identical. This condition may be violated during early human development if amniotic fluid volume is deficient, as the resulting mechanical pressures may increase asymmetries. Indeed, we showed that limb bones of deceased human fetuses exhibited increased asymmetry, when there was not sufficient amniotic fluid (and, thus, space in the uterine cavity. As amniotic fluid deficiency is known to cause substantial asymmetries and abnormal limb development, these subtle asymmetries are probably at least in part caused by the mechanical pressures. On the other hand, deficiencies in amniotic fluid volume are known to be associated with other congenital abnormalities that may disturb DI. More specifically, urogenital abnormalities can directly affect/reduce amniotic fluid volume. We disentangled the direct mechanical effects on FA from the indirect effects of urogenital abnormalities, the latter presumably representing DI. We discovered that both factors contributed significantly to the increase in FA. However, the direct mechanical effect of uterine pressure, albeit statistically significant, appeared less important than the effects of urogenital abnormalities, with an effect size only two-third as large. We, thus, conclude that correcting for the relevant direct factors allowed for a representative test of the association between DI and stress, and confirmed that fetuses form a suitable model system to increase our understanding in patterns of FA and symmetry development.

  11. Value of amniotic fluid IL-8 and Annexin A2 in prediction of preterm delivery in preterm labor and preterm premature rupture of membranes.

    Science.gov (United States)

    Jia, Xiaohui

    2014-01-01

    To investigate the clinical significance and value in the prediction of preterm delivery of combined amniotic fluid IL-8 and Annexin A2 levels in preterm premature rupture of membranes (PPROM) and preterm labor (PTL). Sixty pregnant women at < 32 gestational weeks who developed PTL were divided into a PPROM group and a non-PPROM group. Ten normal pregnant women served as a control group. IL-8 and Annexin A2 levels were measured in amniotic fluid samples from each patient. Amniotic fluid IL-8 and Annexin-A2 levels in PTL (PPROM and non-PPROM groups) were significantly higher than those of the controls (p < 0.05). The PPROM group displayed higher amniotic fluid Annexin-A2 levels than did the non-PPROM group, with a statistically significant difference (p < 0.05). The PPROM group showed higher amniotic fluid IL-8 levels than did the non-PPROM group; however, this was statistically insignificant (p = 0.56). Combined detection of amniotic fluid IL-8 and Annexin-A2 in the prediction of preterm delivery within 2 weeks of measurement showed sensitivity of 81.25%, specificity of 88.89% and PPV of 92.86%. Amniotic fluid IL-8 and Annexin-A2 levels are associated with the occurrence of PPROM and PTL. Combined detection of IL-8 and Annexin-A2 levels in identifying preterm delivery within 2 weeks in PTL and PPROM is of possible clinical and predictive value.

  12. 'Perinatal outcome in preterm premature rupture of membranes with Amniotic fluid index < 5 (AFI < 5

    Directory of Open Access Journals (Sweden)

    Hantoushzadeh Sedigheh

    2004-08-01

    Full Text Available Abstract Background Our purpose was to determine whether AFI Methods We performed a prospective cohort study of 95 singleton pregnancies complicated by preterm premature rupture of the membranes (PPROM with delivery between 26 and 34 weeks gestation. Patients were categorized in two groups on the basis of amniotic fluid index2 and Fisher exact tests. Continuous data were evaluated for normal distribution and tested for significance with the student t test. All 2-sided p values Results Both groups were similar with respect to selected demographics, gestational age at rupture of the membranes, gestational age at the delivery, birth weight. Both groups were similar with respect to selected variable, latency until delivery, early onset neonatal sepsis, RDS and neonatal death. Patients with AFI Conclusions An AFI

  13. Meconium "aspiration" (or respiratory distress associated with meconium-stained amniotic fluid?).

    Science.gov (United States)

    Vain, Nestor E; Batton, Daniel G

    2017-08-01

    The designation meconium aspiration syndrome (MAS) reflects a spectrum of disorders in infants born with meconium-stained amniotic fluid, ranging from mild tachypnea to severe respiratory distress and significant mortality. The frequency of MAS is highest among infants with post-term gestation, thick meconium, and birth asphyxia. Pulmonary hypertension is an important component in severe cases. Prenatal hypopharyngeal suctioning and postnatal endotracheal intubation and suctioning of vigorous infants are not effective. Intubation and suctioning of non-breathing infants is controversial and needs more investigation. Oxygen, mechanical ventilation, and inhaled nitric oxide are the mainstays of treatment. Surfactant is often used in infants with severe parenchymal involvement. High-frequency ventilation and extracorporeal membrane oxygenation are usually considered rescue therapies. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Amniotic fluid-derived mesenchymal stem cells as a novel ...

    African Journals Online (AJOL)

    CLEMENTINA

    2012-06-28

    Jun 28, 2012 ... Apparent structural disorder of liver lobules was detected in the physiological saline (PS) group, disorder hepatic strands (2) and central vein (1). (B) Increased infiltration of mononuclear cells, vacuolar degeneration, and necrosis were observed in the PS group, focal area of hepatic necrosis with dark.

  15. Amniotic fluid-derived mesenchymal stem cells as a novel ...

    African Journals Online (AJOL)

    AFMSCs were transplanted into injured liver via the portal vein in the rat FHF model. Therapeutic effect was evaluated after cell transfusion by histologic pathology, hepatic enzyme levels and animal survival. Cryostat sections were prepared and directly assessed for green fluorescent protein (GFP) expression and ...

  16. Fetoplacental Discrepancy with Normal Karyotype in Amniotic Fluid and Two Different Cell Lines in Placenta

    Directory of Open Access Journals (Sweden)

    Veronica Ortega

    2013-01-01

    Full Text Available We present a case of fetoplacental discrepancy in a second-trimester fetus with normal karyotype in amniotic fluid and two different Robertsonian translocations in placenta. A 41-year-old woman of Middle-Eastern origin, gravida 2, para 1, underwent amniocentesis at 16-week gestation because of advanced maternal age. Amniotic fluid karyotype showed a normal 46,XX karyotype with a homozygous inv(9. Parental chromosome analysis showed both parents to be carriers of inv(9 and the parents are not consanguineous. Fetal ultrasound was normal. The mother presented to the clinic 4 weeks later with intrauterine fetal demise. Chromosome analysis from the placenta showed two different cell lines: a balanced (15;21 Roberstonian translocation in 11 cells and an unbalanced (21;21 Robertsonian translocation in 9 cells. The karyotype was interpreted as mos 45,XX,inv(9(p11q13x2,der(15;21(q10;q10[11]/46,XX,inv(9(p11q13x2,+21,der(21;21(q10;q10. Mother was a carrier for the Cystic Fibrosis (delta F508, Factor V Leiden mutations, HbD-Los Angeles and HbQ-India variants. She also had a sibling with term stillbirth. Her husband’s history was unremarkable. Our case appears to be another example of confined placental mosaicism (CPM with normal fetal karyotype. However, we could not confirm the possibility that CPM contributed to the IUFD in our case given the complex medical history of the mother.

  17. Identification of Symptomatic Fetuses Infected with Cytomegalovirus Using Amniotic Fluid Peptide Biomarkers.

    Directory of Open Access Journals (Sweden)

    Cyrille Desveaux

    2016-01-01

    Full Text Available Cytomegalovirus (CMV is the most common cause of congenital infection, and is a major cause of sensorineural hearing loss and neurological disabilities. Evaluating the risk for a CMV infected fetus to develop severe clinical symptoms after birth is crucial to provide appropriate guidance to pregnant women who might have to consider termination of pregnancy or experimental prenatal medical therapies. However, establishing the prognosis before birth remains a challenge. This evaluation is currently based upon fetal imaging and fetal biological parameters, but the positive and negative predictive values of these parameters are not optimal, leaving room for the development of new prognostic factors. Here, we compared the amniotic fluid peptidome between asymptomatic fetuses who were born as asymptomatic neonates and symptomatic fetuses who were either terminated in view of severe cerebral lesions or born as severely symptomatic neonates. This comparison allowed us to identify a 34-peptide classifier in a discovery cohort of 13 symptomatic and 13 asymptomatic neonates. This classifier further yielded 89% sensitivity, 75% specificity and an area under the curve of 0.90 to segregate 9 severely symptomatic from 12 asymptomatic neonates in a validation cohort, showing an overall better performance than that of classical fetal laboratory parameters. Pathway analysis of the 34 peptides underlined the role of viral entry in fetuses with severe brain disease as well as the potential importance of both beta-2-microglobulin and adiponectin to protect the injured fetal brain infected with CMV. The results also suggested the mechanistic implication of the T calcium channel alpha-1G (CACNA1G protein in the development of seizures in severely CMV infected children. These results open a new field for potential therapeutic options. In conclusion, this study demonstrates that amniotic fluid peptidome analysis can effectively predict the severity of congenital CMV

  18. Nukbone® promotes proliferation and osteoblastic differentiation of mesenchymal stem cells from human amniotic membrane

    Energy Technology Data Exchange (ETDEWEB)

    Rodríguez-Fuentes, Nayeli; Rodríguez-Hernández, Ana G. [Depto. Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Mexico City 04510 (Mexico); Enríquez-Jiménez, Juana [Depto. Biología de la Reproducción, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán (INCMNSZ), México City 14000 (Mexico); Alcántara-Quintana, Luz E. [Subd. de Investigación, Centro Nacional de la Transfusión Sanguínea, Secretaria de Salud, Mexico City 07370 (Mexico); Fuentes-Mera, Lizeth [Depto. Biología Molecular e Histocompatibilidad, Hospital General “Dr. Manuel Gea González”, México City 4800 (Mexico); Piña-Barba, María C. [Depto. Materiales Metálicos y Cerámicos, Instituto de Investigaciones en Materiales, Universidad Nacional Autónoma de México (UNAM), México City 04510 (Mexico); Zepeda-Rodríguez, Armando [Depto. Biología Celular y Tisular, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), México City 04510 (Mexico); and others

    2013-05-10

    Highlights: •Nukbone showed to be a good scaffold for adhesion, proliferation and differentiation of stem cells. •Nukbone induced osteoblastic differentiation of human mesenchymal stem cells. •Results showed that Nukbone offer an excellent option for bone tissue regeneration due to properties. -- Abstract: Bovine bone matrix Nukbone® (NKB) is an osseous tissue-engineering biomaterial that retains its mineral and organic phases and its natural bone topography and has been used as a xenoimplant for bone regeneration in clinics. There are not studies regarding its influence of the NKB in the behavior of cells during the repairing processes. The aim of this research is to demonstrate that NKB has an osteoinductive effect in human mesenchymal stem cells from amniotic membrane (AM-hMSCs). Results indicated that NKB favors the AM-hMSCs adhesion and proliferation up to 7 days in culture as shown by the scanning electron microscopy and proliferation measures using an alamarBlue assay. Furthermore, as demonstrated by reverse transcriptase polymerase chain reaction, it was detected that two gene expression markers of osteoblastic differentiation: the core binding factor and osteocalcin were higher for AM-hMSCs co-cultured with NKB in comparison with cultivated cells in absence of the biomaterial. As the results indicate, NKB possess the capability for inducing successfully the osteoblastic differentiation of AM-hMSC, so that, NKB is an excellent xenoimplant option for repairing bone tissue defects.

  19. Comprehensive proteomic analysis of the human amniotic fluid proteome: gestational age-dependent changes.

    Science.gov (United States)

    Michaels, John-Edward A; Dasari, Surendra; Pereira, Leonardo; Reddy, Ashok P; Lapidus, Jodi A; Lu, Xinfang; Jacob, Thomas; Thomas, Archana; Rodland, Matthew; Roberts, Charles T; Gravett, Michael G; Nagalla, Srinivasa R

    2007-04-01

    Amniotic fluid (AF) is a significant contributor to fetal health and constitutes a potential rich source of biomarkers for diagnosis of maternal and fetal disorders. In this study, we performed a comprehensive survey of the proteins expressed in AF, combining gel and liquid-based fractionation approaches coupled with LC-MS/MS analysis. Two-dimensional Liquid Chromatography (2D-LC) analysis identified 118 nonredundant proteins with high confidence. One- and two-dimensional gel electrophoresis and in-gel digestion identified 101 proteins. Combining both sets resulted in 219 proteins, of which 96 are unique to AF; 70, 18, and 35 proteins are present in serum, cervico-vaginal fluid, and all three fluids, respectively. Fluorescence two-dimensional differential in-gel electrophoresis (2D-DIGE) comparison of first-, second-, and third-trimester AF samples revealed that maximal differences in the relative abundance of AF proteins occur between the first and second trimesters. A systematic analysis of proteins present both in AF and maternal serum could lead to the development of new noninvasive diagnostic procedures to monitor fetal status.

  20. Amniotic fluid CA-125 as a marker of intra-amniotic inflammation associated with preterm delivery: a preliminary single center study.

    Science.gov (United States)

    Seong, Won Joon

    2016-01-01

    The purpose of this study was to investigate whether amniotic fluid (AF) CA-125 in patients with preterm labor or preterm premature rupture of membranes can help predict intra-amniotic inflammation (IAI), microbial invasion of the amniotic cavity (MIAC) and imminent delivery. We recruited 36 women who admitted with impending preterm delivery and suspicious AF infection. AF matrix metalloproteinase-8 (MMP-8), white blood cell (WBC) count, glucose levels, and CA-125 levels were measured, and the MMP-8 bedside rapid test was also performed. AF culture and PCR were subsequently performed to confirm MIAC. We compared AF CA-125 levels according to the presence of IAI or MIAC and assessed its predictive value for delivery within 7 days of admission. AF CA-125 levels were significantly higher in the IAI group than in the non-IAI group (mean ± standard deviation: 5608 ± 864 vs 904 ± 84 IU/ml; p = 0.001). AF CA-125 levels showed a negative correlation with gestational age and a positive correlation with AF WBC counts and MMP-8 levels. AF CA-125 levels were higher in the MIAC group, though this difference was not statistically significant (p = 0.064). Delivery within 7 days of admission was significantly more common in patients with higher AF CA-125 levels (cut-off: 1650 IU/ml, sensitivity: 71.4 %, specificity: 86.4 %, p = 0.005). AF CA-125 levels are increased in patients with AF inflammation and can be a predictor of imminent preterm delivery.

  1. Elemental analysis of human amniotic fluid and placenta by TXRF and EDXRF: child weight and aging mother dependences

    International Nuclear Information System (INIS)

    Carvalho, M.L.; Custodio, P.J.; Reus, U.; Prange, A.

    2000-01-01

    This work is an attempt to evaluate the possible influence of the age's mother in trace elemental concentration in human amniotic fluid and placenta and whether these concentrations are correlated to the weight of the newborn infants. Total reflection x-ray fluorescence (TXRF) was used to analyze 16 amniotic fluid samples, and the placenta samples were analyzed by energy dispersive x-ray fluorescence (EDXRF). The whole samples were collected during the delivery, from healthy mothers and healthy infants. According to the age of the mother three different groups were considered: from 20-25, 25-30 and 30-40. Only two mothers were aged more than 35. The weight of the infants ranged from 2.56 to 4.05 kg. The organic matrices of the amniotic fluid samples were removed by wet ashing with HNO 3 in oxygen plasma. Yttrium was used as internal standard, for TXRF analysis. For EDXRF placenta samples were lyophilized and analyzed without any chemical treatment. Very low levels of Ni, and Sr were found in the amniotic fluid samples, independently of age of the mother and child weight. Cr, Mn, Se and Pb were at the level of the detection limit. Zn, considered one of the key elements in the newborn health, was not significantly different in the analyzed samples, however, was related, though weakly, to birth weigh. The obtained concentrations ranged from 0.11 to 0.92 mg/L and 30 to 65 μg/g in amniotic fluid and placenta respectively. The only two elements seemed to be significantly affected with age mother and newborn weight were Ca and Fe for both kind of samples: Ca levels were increased in heavier children and elder mothers, however Fe increased with the increase of the age mother but decreased for heavier babies. The same conclusions were obtained for placenta and amniotic fluid samples. Cu is closely associated in its function in the organism with Fe and has a similar behavior with this element, however not so pronounced. (author)

  2. Proteomic profiling of amniotic fluid in preterm labor using two-dimensional liquid separation and mass spectrometry.

    Science.gov (United States)

    Bujold, Emmanuel; Romero, Roberto; Kusanovic, Juan Pedro; Erez, Offer; Gotsch, Francesca; Chaiworapongsa, Tinnakorn; Gomez, Ricardo; Espinoza, Jimmy; Vaisbuch, Edi; Mee Kim, Yeon; Edwin, Samuel; Pisano, Mike; Allen, Beth; Podust, Vladimir N; Dalmasso, Enrique A; Rutherford, Jennifer; Rogers, Wade; Moser, Allan; Yoon, Bo Hyun; Barder, Tim

    2008-10-01

    Simultaneous analysis of the protein composition of biological fluids is now possible. Such an approach can be used to identify biological markers of disease and to understand the pathophysiology of disorders that have eluded classification, diagnosis, and treatment. The purpose of this study was to analyze the differences in protein composition of the amniotic fluid of patients in preterm labor. Amniotic fluid was obtained by amniocentesis from three groups of women with preterm labor and intact membranes: (1) women without intra-amniotic infection/inflammation (IAI) who delivered at term, (2) women without IAI who delivered a preterm neonate, and (3) women with IAI. Intra-amniotic infection was defined as a positive amniotic fluid culture for microorganisms. Intra-amniotic inflammation was defined as an elevated amniotic fluid interleukin (IL)-6 (> or =2.3 ng/mL). Two-dimensional (2D) chromatography was used for analysis. The first dimension separated proteins by isoelectric point, while the second, by the degree of hydrophobicity. 2D protein maps were generated using different experimental conditions (reducing agents as well as protein concentration). The maps were used to discern subsets of isoelectric point/hydrophobicity containing differentially expressed proteins. Protein identification of differentially expressed fractions was conducted with mass spectrometry. Enzyme-linked immunosorbent assays (ELISA) as well as surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS)-based on-chip antibody capture immunoassays were also used for confirmation of a specific protein that was differentially expressed. (1) Amniotic fluid protein composition can be analyzed using a combination of 2D liquid chromatography and mass spectrometry for the identification of proteins differentially expressed in patients in preterm labor. (2) While total insulin-like growth factor-binding protein-1 (IGFBP-1) concentration did not change, IGFBP-1

  3. Secreted phospholipase A2 is increased in meconium-stained amniotic fluid of term gestations: potential implications for the genesis of meconium aspiration syndrome.

    Science.gov (United States)

    Romero, Roberto; Yoon, Bo Hyun; Chaemsaithong, Piya; Cortez, Josef; Park, Chan-Wook; Gonzalez, Rogelio; Behnke, Ernesto; Hassan, Sonia S; Gotsch, Francesca; Yeo, Lami; Chaiworapongsa, Tinnakorn

    2014-07-01

    Meconium-stained amniotic fluid (MSAF) represents the passage of fetal colonic content into the amniotic cavity. Meconium aspiration syndrome (MAS) is a complication that occurs in a subset of infants with MSAF. Secreted phospholipase A2 (sPLA2) is detected in meconium and is implicated in the development of MAS. The purpose of this study was to determine if sPLA2 concentrations are increased in the amniotic fluid of women in spontaneous labor at term with MSAF. This was a cross-sectional study of patients in spontaneous term labor who underwent amniocentesis (n = 101). The patients were divided into two study groups: (1) MSAF (n = 61) and (2) clear fluid (n = 40). The presence of bacteria and endotoxin as well as interleukin-6 (IL-6) and sPLA2 concentrations in the amniotic fluid were determined. Statistical analyses were performed to test for normality and bivariate analysis. The Spearman correlation coefficient was used to study the relationship between sPLA2 and IL-6 concentrations in the amniotic fluid. Patients with MSAF have a higher median sPLA2 concentration (ng/mL) in amniotic fluid than those with clear fluid [1.7 (0.98-2.89) versus 0.3 (0-0.6), p meconium is aspirated before birth.

  4. Modulation of intestinal inflammation by minimal enteral nutrition with amniotic fluid in preterm pigs

    DEFF Research Database (Denmark)

    Østergaard, Mette V; Bering, Stine Brandt; Jensen, Michael L

    2014-01-01

    Background: Necrotizing enterocolitis (NEC) is a severe inflammatory disorder, associated with the difficult transition from parenteral to enteral feeding after preterm birth. We hypothesized that minimal enteral nutrition (MEN) with amniotic fluid (AF), prior to enteral formula feeding, would...... improve resistance to NEC in preterm pigs. Methods: Experiment 1: IEC-6 cells were incubated with porcine (pAF) and human AF (hAF) to test AF-stimulated enterocyte proliferation and migration in vitro. Experiment 2: Cesarean-delivered, preterm pigs were fed parenteral nutrition and MEN with pAF, h......AF, or control fluid (MEN-pAF, MEN-hAF, or MEN-CTRL; all n = 9) for 2 days before tissue collection. Experiment 3: Preterm pigs were fed MEN diets as in experiment 2, but followed by 2 days of enteral formula feeding, which predisposes to NEC (NEC-pAF, NEC-hAF, or NEC-CTRL; n = 10–12). Results: Both pAF and h...

  5. Concentrations of cobalt, rubidium, selenium and zinc in maternal and cord blood serum and amniotic fluid of women with normal and prolonged pregnancies

    International Nuclear Information System (INIS)

    Antoniou, K.; Lolis, D.; Vassilaki-Grimani, M.; Grimanis, A.P.

    1982-01-01

    Instrumental neutron activation analysis has been applied for the determination of Co, Rb, Se and Zn in maternal and umbilical cord serum and amniotic fluid of women with normal pregnancy and prolonged pregnancy. Significantly lower levels of Co, Se and Zn were found in maternal blood serum and cord serum of women with prolonged pregnancy as compared with those in sera of mothers with normal pregnancy. Zinc concentrations were also found significantly lower in amniotic fluids of women with prolonged pregnancy. (author)

  6. Maternal-Fetal Transfer and Amniotic Fluid Accumulation of Nucleoside Analogue Reverse Transcriptase Inhibitors in Human Immunodeficiency Virus-Infected Pregnant Women

    OpenAIRE

    Chappuy, Hélène; Tréluyer, Jean-Marc; Jullien, Vincent; Dimet, Jérôme; Rey, Elisabeth; Fouché, Maria; Firtion, Ghislaine; Pons, Gérard; Mandelbrot, Laurent

    2004-01-01

    This study was performed to investigate placental transfer of nucleoside analogue reverse transcriptase inhibitors (NRTIs) and their concentrations in amniotic fluid when given to human immunodeficiency virus (HIV)-infected pregnant women. A total of 100 HIV type 1-infected mothers receiving antiretroviral therapy, including one or more NRTIs, for clinical indications at the time of delivery were enrolled. Maternal blood samples and amniotic fluid were obtained during delivery or cesarean sec...

  7. Human umbilical cord mesenchymal stem cell-loaded amniotic membrane for the repair of radial nerve injury.

    Science.gov (United States)

    Li, Zhi; Qin, Hanjiao; Feng, Zishan; Liu, Wei; Zhou, Ye; Yang, Lifeng; Zhao, Wei; Li, Youjun

    2013-12-25

    In this study, we loaded human umbilical cord mesenchymal stem cells onto human amniotic membrane with epithelial cells to prepare nerve conduits, i.e., a relatively closed nerve regeneration chamber. After neurolysis, the injured radial nerve was enwrapped with the prepared nerve conduit, which was fixed to the epineurium by sutures, with the cell on the inner surface of the conduit. Simultaneously, a 1.0 mL aliquot of human umbilical cord mesenchymal stem cell suspension was injected into the distal and proximal ends of the injured radial nerve with 1.0 cm intervals. A total of 1.75 × 10(7) cells were seeded on the amniotic membrane. In the control group, patients received only neurolysis. At 12 weeks after cell transplantation, more than 80% of patients exhibited obvious improvements in muscular strength, and touch and pain sensations. In contrast, these improvements were observed only in 55-65% of control patients. At 8 and 12 weeks, muscular electrophysiological function in the region dominated by the injured radial nerve was significantly better in the transplantation group than the control group. After cell transplantation, no immunological rejections were observed. These findings suggest that human umbilical cord mesenchymal stem cell-loaded amniotic membrane can be used for the repair of radial nerve injury.

  8. Human umbilical cord mesenchymal stem cell-loaded amniotic membrane for the repair of radial nerve injury

    Science.gov (United States)

    Li, Zhi; Qin, Hanjiao; Feng, Zishan; Liu, Wei; Zhou, Ye; Yang, Lifeng; Zhao, Wei; Li, Youjun

    2013-01-01

    In this study, we loaded human umbilical cord mesenchymal stem cells onto human amniotic membrane with epithelial cells to prepare nerve conduits, i.e., a relatively closed nerve regeneration chamber. After neurolysis, the injured radial nerve was enwrapped with the prepared nerve conduit, which was fixed to the epineurium by sutures, with the cell on the inner surface of the conduit. Simultaneously, a 1.0 mL aliquot of human umbilical cord mesenchymal stem cell suspension was injected into the distal and proximal ends of the injured radial nerve with 1.0 cm intervals. A total of 1.75 × 107 cells were seeded on the amniotic membrane. In the control group, patients received only neurolysis. At 12 weeks after cell transplantation, more than 80% of patients exhibited obvious improvements in muscular strength, and touch and pain sensations. In contrast, these improvements were observed only in 55–65% of control patients. At 8 and 12 weeks, muscular electrophysiological function in the region dominated by the injured radial nerve was significantly better in the transplantation group than the control group. After cell transplantation, no immunological rejections were observed. These findings suggest that human umbilical cord mesenchymal stem cell-loaded amniotic membrane can be used for the repair of radial nerve injury. PMID:25206667

  9. Amniotic fluid concentrations of soluble scavenger receptor for hemoglobin (sCD163) in pregnancy complicated by preterm premature rupture of the membranes and histologic chorioamnionitis.

    Science.gov (United States)

    Kacerovsky, Marian; Drahosova, Marcela; Andrys, Ctirad; Hornychova, Helena; Tambor, Vojtech; Lenco, Juraj; Tosner, Jindrich; Krejsek, Jan

    2011-08-01

    To determine changes in the amniotic fluid, soluble form of scavenger receptor for hemoglobin (sCD163) concentrations during advancing gestation, and in patients with preterm premature rupture of membranes (PPROM) complicated by histological chorioamnionitis were studied. One hundred and fifty-two women with singleton pregnancies were enrolled. The concentration of sCD163 in amniotic fluid was determined using sandwich enzyme immunoassay technique. Women in the midtrimester had a significantly higher median amniotic fluid sCD163 concentration than those at term not in labor (308 ng/ml vs. 217 ng/ml; p = 0.04). Patients with PPROM and histological chorioamnionitis had a higher median amniotic fluid sCD163 level than those with PPROM without histological chorioamnionitis (885 ng/ml vs. 288 ng/ml; p < 0.0001). Amniotic fluid sCD163 concentrations decrease with advancing gestation. Amniotic fluid sCD163 concentrations are significantly higher in women with PPROM between 24 and 36 gestational weeks with histological chorioamnionitis than those without histological signs of inflammation.

  10. Gastric lavage for prevention of feeding problems in neonates with meconium-stained amniotic fluid: a randomised controlled trial.

    Science.gov (United States)

    Sharma, Preeti; Nangia, Sushma; Tiwari, Soumya; Goel, Ankita; Singla, Bhupesh; Saili, Arvind

    2014-05-01

    The role of gastric lavage in preventing retching, vomiting and secondary meconium aspiration syndrome in neonates with meconium-stained amniotic fluid is uncertain, and no there are no definitive guidelines. To evaluate the effect of gastric lavage in preventing retching, vomiting and secondary meconium aspiration syndrome in neonates with meconium-stained amniotic fluid. This was an open-label, parallel, randomized controlled trial conducted in the labour room, postnatal and neonatal wards of a tertiary-care teaching hospital. Vigorous neonates of ≧34 weeks gestation with meconium-stained amniotic fluid were randomised into two groups using block randomisation. Infants requiring oxygen, in respiratory distress or with major congenital malformations were excluded. Infants in the study group received elective gastric lavage in the labour room after initial stabilisation. No gastric lavage was done in the control group. The newborns were assessed for retching, vomiting and secondary meconium aspiration syndrome in the first 48 hrs of life or until discharge from the hospital, whichever was later. A total of 267 newborns were randomly assigned to the gastric lavage group and 269 to the no gastric lavage group. There were no statistical differences in overall feeding between the two groups (6·74% vs 10·78%). Feeding of two newborns in the no-lavage group had to be omitted for the initial few hours because of vomiting; this did not happen in any newborn in the lavage group. No newborn in either group developed secondary meconium aspiration syndrome. Gastric lavage in newborns with meconium-stained amniotic fluid does not prevent or reduce the occurrence of feeding problems or secondary meconium aspiration syndrome.

  11. Racial disparity in meconium-stained amniotic fluid and meconium aspiration syndrome in the United States, 1989-2000.

    Science.gov (United States)

    Sriram, Sudhir; Wall, Stephen N; Khoshnood, Babak; Singh, Jaideep K; Hsieh, Hui-Lung; Lee, Kwang-Sun

    2003-12-01

    To estimate the prevalence of meconium-stained amniotic fluid and meconium aspiration syndrome, as well as the differences in case fatality from meconium aspiration syndrome, between non-Hispanic black and non-Hispanic white infants. We studied non-Hispanic black and non-Hispanic white live births with weights greater than 2.5 kg and gestational ages greater than 35 weeks, using the linked US birth and infant death cohorts for three periods: 1989-1991, 1995-1997, and 1998-2000. We used logistic regression to estimate the risks of meconium-stained amniotic fluid and meconium aspiration syndrome and to estimate the case fatality of meconium aspiration syndrome by maternal race, birth weight, period, and pregnancy complications. Risk of meconium-stained amniotic fluid was 80% higher in non-Hispanic blacks when compared with non-Hispanic whites (birth weight-adjusted odds ratio [OR], 1.81, 95% confidence interval [CI] 1.80, 1.82). The prevalence of pregnancy complications did not explain this racial disparity. Risk of meconium aspiration syndrome in non-Hispanic blacks was 67% higher when compared with non-Hispanic whites (birth weight-adjusted OR 1.67, 95% CI 1.64, 1.70). The case fatality rate of meconium aspiration syndrome was similar between non-Hispanic blacks and non-Hispanic whites in the three periods, with rates of 15.5, 15.2, and 11.2 per 1000 in non-Hispanic blacks and 13.5, 11.2, and 10.1 per 1000 in non-Hispanic whites in 1989-1991, 1995-1997, and 1998-2000, respectively. Our results suggest that when compared with non-Hispanic whites, non-Hispanic blacks are at significantly greater risk for meconium-stained amniotic fluid and meconium aspiration syndrome but not for meconium aspiration syndrome case fatality.

  12. Meconium-Stained Amniotic Fluid as an Independent Risk Factor for Fever and Postpartum Infection in Term Pregnancy

    Directory of Open Access Journals (Sweden)

    M Valadan

    2008-06-01

    Full Text Available Background: The aim of this study was to statistically evaluate the hypothesis that the presence of meconium-stained amniotic fluid is associated with postpartum maternal infection.Methods: This prospective cohort study included 573 term pregnant women in labor, with no other medical problems, that underwent cesarean section for pregnancy termination. Women with prolonged active-phase labor, prolonged rupture of membranes, complicated cesarean section and pre-operative infections were excluded from this study.The subjects were divided into two groups: 280 women with meconium-stained amniotic fluid (M group, and 293 women with clear amniotic fluid (C group. A comparison was done regarding postpartum fever, endometritis and wound infection between the two groups. Students t-test and chi square test were used for data analysis, along with linear regression, with p<0.05 indicating significance.Results: Among the 573 women, a total of 82 women (14% had fever after cesarean; 42 women from the M group, and 40 women from the C group (p= 0.3. Among the 82 women who had fever, 33 women had continuous fever, 6.5% in group M and 2.7% in group C (p<0.05, RR: 1.98, 95% CI 1.3-3.1. Among the 573 women, 4% developed endometritis 5% in group M and 2% in group C (p<0.05, RR: 2.3, 95%, CI 1.3-3.4. Similarly, among the 573 women, a total of 5 women (1% developed wound infection, 1.7% in group M and 0.68% in group C (p=0.7.Conclusion: Meconium-stained amniotic fluid is associated with increased postpartum infection independent of other risk factors for infection.

  13. Amniotic Fluid Arginine from Gestational Weeks 13 to 15 Is a Predictor of Birth Weight, Length, and Head Circumference

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    Astrid Bjørke-Jenssen

    2017-12-01

    Full Text Available Arginine is a constituent of proteins and a precursor for polyamines and nitric oxide, and is essential for placentation, angiogenesis, and growth. Maternal plasma arginine concentrations are found to be lower in pregnancies complicated by fetal growth restriction, and arginine supplementation in later pregnancy is reported to increase birth weight. We measured arginine and the metabolites asymmetric dimethylarginine (ADMA and symmetric dimethylarginine (SDMA in the amniotic fluid obtained in pregnancy weeks 13 to 15 from 363 pregnancies with a documented normal outcome and related the concentrations to birth weight, length, and head circumference. Arginine was higher in the amniotic fluid from female (mean 40.8 (SD 10.6 µmol/L compared to male fetuses (37.4 (SD 11.2 µmol/L, p = 0.003. Despite the gender difference, arginine in the amniotic fluid from gestational weeks 13–15 was the strongest predictor for birth weight, length, and head circumference. ADMA was a strong predictor for birth weight and length, SDMA for birth weight, while Arg/ADMA and Arg/SDMA only predicted head circumference in multiple linear regression models. Due to increased arginine demands, pregnancy is considered a state of relative arginine deficiency. Our findings reflect the importance of a good maternal arginine status in early pregnancy, an observation that should be evaluated in an intervention study.

  14. The LARGE principle of cellular reprogramming: lost, acquired and retained gene expression in foreskin and amniotic fluid-derived human iPS cells.

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    Katharina Wolfrum

    Full Text Available Human amniotic fluid cells (AFCs are routinely obtained for prenatal diagnostics procedures. Recently, it has been illustrated that these cells may also serve as a valuable model system to study developmental processes and for application in regenerative therapies. Cellular reprogramming is a means of assigning greater value to primary AFCs by inducing self-renewal and pluripotency and, thus, bypassing senescence. Here, we report the generation and characterization of human amniotic fluid-derived induced pluripotent stem cells (AFiPSCs and demonstrate their ability to differentiate into the trophoblast lineage after stimulation with BMP2/BMP4. We further carried out comparative transcriptome analyses of primary human AFCs, AFiPSCs, fibroblast-derived iPSCs (FiPSCs and embryonic stem cells (ESCs. This revealed that the expression of key senescence-associated genes are down-regulated upon the induction of pluripotency in primary AFCs (AFiPSCs. By defining distinct and overlapping gene expression patterns and deriving the LARGE (Lost, Acquired and Retained Gene Expression Principle of Cellular Reprogramming, we could further highlight that AFiPSCs, FiPSCs and ESCs share a core self-renewal gene regulatory network driven by OCT4, SOX2 and NANOG. Nevertheless, these cell types are marked by distinct gene expression signatures. For example, expression of the transcription factors, SIX6, EGR2, PKNOX2, HOXD4, HOXD10, DLX5 and RAXL1, known to regulate developmental processes, are retained in AFiPSCs and FiPSCs. Surprisingly, expression of the self-renewal-associated gene PRDM14 or the developmental processes-regulating genes WNT3A and GSC are restricted to ESCs. Implications of this, with respect to the stability of the undifferentiated state and long-term differentiation potential of iPSCs, warrant further studies.

  15. Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication

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    Natascha Krömmelbein

    2016-02-01

    Full Text Available The human cytomegalovirus (HCMV replicates to high titers in primary human fibroblast cell cultures. A variety of primary human cells and some tumor-derived cell lines do also support permissive HCMV replication, yet at low levels. Cell lines established by transfection of the transforming functions of adenoviruses have been notoriously resistant to HCMV replication and progeny production. Here, we provide first-time evidence that a permanent cell line immortalized by adenovirus type 5 E1A and E1B (CAP is supporting the full HCMV replication cycle and is releasing infectious progeny. The CAP cell line had previously been established from amniotic fluid cells which were likely derived from membranes of the developing fetus. These cells can be grown under serum-free conditions. HCMV efficiently penetrated CAP cells, expressed its immediate-early proteins and dispersed restrictive PML-bodies. Viral DNA replication was initiated and viral progeny became detectable by electron microscopy in CAP cells. Furthermore, infectious virus was released from CAP cells, yet to lower levels compared to fibroblasts. Subviral dense bodies were also secreted from CAP cells. The results show that E1A/E1B expression in transformed cells is not generally repressive to HCMV replication and that CAP cells may be a good substrate for dense body based vaccine production.

  16. Multicenter proficiency study for detection of Toxoplasma gondii in amniotic fluid by nucleic acid amplification methods.

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    Kaiser, Karine; Van Loon, Anton M; Pelloux, Hervé; Ferrandiz, Josette; Picot, Stéphane; Wallace, Paul; Peyron, François

    2007-01-01

    A proficiency panel was designed to assess the performance of nucleic acid amplification technologies for the detection of Toxoplasma gondii in amniotic fluid. The proficiency panel consisted of five lyophilised coded samples in a range of concentration between 5 to 1000 parasites/ml and a negative control. The distribution also included a questionnaire on the applied methods. Thirty-three laboratories in 17 countries participated and returned a total of 38 data sets. The percentage of data sets achieving correct results on all panel samples was 42.1%, whereas two or more incorrect or equivocal results were reported for 36.8%. The lowest concentration (5 parasites/ml) was not identified correctly in 15 (39.5%) data sets. False positive results were reported by two laboratories both of which had not included a step in their procedure to rule out contamination. In 32 (84.2%) data sets an "in-house" method was used, and in 6 (15.8%) sets a commercial assay was applied. Overall, the results of this study demonstrate the need for improvements in both sensitivity and specificity of molecular detection methods of T. gondii and for the development of international reference materials to help laboratories with the development and validation of their assays.

  17. Accuracy Assessment of Interphase Fluorescence In-Situ Hybridization on Uncultured Amniotic Fluid Cells

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    Hamideh Karimi

    2007-01-01

    Full Text Available Background: Parental anxiety while waiting for the results of amniocentesis has been investigatedby many authors. It seems that the implementation of faster techniques such as fluorescence in-situhybridization (FISH will have some benefits in reducing this anxiety. Besides the patients' attitudesto choosing this method, gynecologists who are the persons responsible for treatment, must feelcomfortable about prescribing FISH techniques.Materials and Methods: This study, using a simple methodology, was undertaken to evaluate theresults of FISH tests on the amniotic fluid from 40 pregnant women undergoing cesarean surgery.Two sets of probes including X/Y cocktail and 13, 21 and 18 were applied on different slides.Results: The results of FISH tests were compared with the reports of the pediatrician about thehealth condition of the newborn. Complete conformity between the two sets of findings, haveconvinced our gynecologists of the benefit of prescribing this method to reduce the anxiety ofpatients at risk of having abnormal offspring due to chromosomal anuploidies.Conclusion: As has been documented by many authors, conventional chromosome analysis hasgreat advantages over fluorescence in situ hybridization of interphase amniocytes, but reducing theanxiety of parents is a good reason for employing the FISH technique.

  18. Low amniotic fluid index in high risk pregnancy and poor apgar score at birth

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    Sultana, S.; Akhtar, K.A.K.

    2008-01-01

    To determine the accuracy of antepartum Amniotic Fluid Index (AFI) of 5 cm was labeled as predictor of good outcome at birth. The subjects in both the groups were demographically matched and fulfilled the inclusion and exclusion criteria. The Apgar score was calculated at 5 minutes of birth. The newborns, with Apgar score 6 were labeled as healthy. AFI was compared with Apgar score, using Chi-square and a p-value was calculated to determine the statistical significance. Sensitivity, specificity, efficiency and the predictive values of AFI at a cut off point of < 5 cm as a predictor of adverse outcome at birth (Apgar score of < 6 at 5 minutes of birth) in high-risk pregnancy were calculated. Only 8 neonates of 50 women with low AFI had low Apgar score. Similarly, 6 neonates of 50 women with normal AFI had poor Apgar score. The diagnostic sensitivity, specificity, positive predictive value, negative predictive value and efficiency of AFI as test were 57.1%, 51.3%, 16%, 88% and 52% respectively. Low AFI is a poor predictor of adverse outcome for high-risk term patients. AFI is not a good screening test for high-risk pregnant women at term for birth of an infant with low Apgar score. (author)

  19. Amniotic fluid index and estimated fetal weight for prediction of fetal macrosomia: a prospective observational study.

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    El Khouly, Nabih I; Elkelani, Osama A; Saleh, Said A

    2017-08-01

    The purpose of this study was to assess the value of combining the estimated fetal weight (EFW) and amniotic fluid index (AFI) measured in term patients early in labor with intact membranes for prediction of macrosomia. In a single center, prospective observational study, 600 patients in the first stage of labor before rupture of membranes in whom ultrasonography was performed to measure AFI and EFW, and these data were analyzed statistically to evaluate prediction of fetal macrosomia. Macrosomia occurred in 64 cases (10.6%). The AFI was significantly higher in the macrosomic group (p = 0.001). It was noted that the area under receiver operating characteristic (ROC) curves for EFW was 0.93 and that of AFI was 0.67. Based on suggested combined EFW and AFI cutoffs of 4000 g and 164 mm, respectively, the positive predictive value (PPV) for combined parameters (92.3%) was higher than that of EFW (75%) and that of AFI (27%) and the likelihood ratio for combination (93.7%) was higher than that of EFW (24.7%) and that of AFI (21%). Combined use of EFW and AFI improves prediction of macrosomia at birth rather than the EFW alone.

  20. Polyhydramnios - frequency of congenital anomalies in relation to the value of the amniotic fluid index.

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    Kornacki, Jakub; Adamczyk, Magdalena; Wirstlein, Przemysław; Osiński, Maciej; Wender-Ożegowska, Ewa

    2017-01-01

    The aims of our study were to assess the correlation between the amniotic fluid index (AFI) value and the frequency and type of fetal anomalies. The material included 94 patients at the third trimester of pregnancy, 60 with mild polyhydramnios, 19 with moderate one, and 15 with severe one. Polyhydramnios was diagnosed if AFI was > 24 cm. All patients were divided into three groups based on the value of AFI: 1) mild polyhydramnios with AFI between 24.1 and 29.9 cm, 2) moderate polyhydramnios with AFI between 30-34.9 cm, and 3) severe polyhydramnios with AFI ≥ 35 cm. The incidence of fetal malformations correlated significantly with the degree of polyhydramnios and was the highest in patients with severe polyhydramnios (53.3%, p = 0.002). Congenital malformations of the gastrointestinal tract were the most frequent fetal anomalies in the whole group of patients (5.3%). Trisomy 18 was the most frequent aneuploidy found in women with polyhydramnios (2.1%). The incidence of fetal congenital anomalies significantly increases with the degree of polyhydramnios, being most frequent in severe one and rather rare in a mild one. Congenital malformations of the gastrointestinal tract were the most frequent anomalies in patients with polyhy-dramnios, especially in women with severe polyhydramnios.

  1. Maximal amniotic fluid index as a prognostic factor in pregnancies complicated by polyhydramnios.

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    Pri-Paz, S; Khalek, N; Fuchs, K M; Simpson, L L

    2012-06-01

    Polyhydramnios is present in approximately 2% of pregnancies and has been associated with a variety of adverse pregnancy outcomes. Our aim was to evaluate the association between the maximal amniotic fluid index (AFI) and the frequency of specific adverse outcomes. This was a retrospective chart review of 524 singleton pregnancies diagnosed with polyhydramnios and delivered in a single tertiary referral center between 2003 and 2008. Polyhydramnios was defined as either AFI ≥ 25 cm or a maximum vertical pocket (MVP) ≥ 8 cm even in the presence of AFI polyhydramnios, as based on the maximal AFI (≥ 35 cm; n = 67), was associated with the highest rates of prenatally diagnosed congenital anomalies (79%), preterm delivery (46%), small-for-gestational-age neonate (16%), aneuploidy (13%) and perinatal mortality (27%). No significant association between degree of polyhydramnios and adverse outcome was demonstrated in cases of idiopathic polyhydramnios (n = 253). There is an association between the frequencies of a variety of adverse pregnancy outcomes and the severity of polyhydramnios as reflected by the maximal AFI. Copyright © 2012 ISUOG. Published by John Wiley & Sons, Ltd.

  2. BDNF Val66Met polymorphism and protein levels in Amniotic Fluid

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    Calabrese Francesca

    2010-02-01

    Full Text Available Abstract Background Brain-Derived Neurotrophic Factor (BDNF is a neurotrophin which plays survival- and growth-promoting activity in neuronal cells and it is involved in cellular plasticity mechanisms as it controls activity dependent synaptic transmission. A functional polymorphism (Val66Met in the pro-region of BDNF, which affects the intracellular trafficking of proBDNF has been associated with memory and cognitive deficits as well as to an increased susceptibility for several psychiatric disorders especially those with a neurodevelopmental origin. To date, no study has evaluated the influence of the Val66Met polymorphism on BDNF levels in a peripheral system that may reflect fetal neurodevelopment. Therefore we investigated in amniotic fluids (AF obtained from 139 healthy women during 15-17 week of pregnancy, BDNF protein levels in correlation with the Val66Met polymorphism. Results Interestingly we found a significant BDNF protein levels reduction in 55 Met carriers (Val/Met and Met/Met (p = 0.002 as compared to 84 non carriers (Val/Val, and no effect of fetus gender, maternal age or gestation week on BDNF levels has been observed. Conclusion These results, although explorative, indicate that during fetal life the Val66Met genotype might influences BDNF protein levels in AF supporting the involvement of this polymorphism in behavioral and functional brain individual differences in the adulthood.

  3. Metabolite profiling of human amniotic fluid by hyphenated nuclear magnetic resonance spectroscopy.

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    Graça, Gonçalo; Duarte, Iola F; J Goodfellow, Brian; Carreira, Isabel M; Couceiro, Ana Bela; Domingues, Maria do Rosário; Spraul, Manfred; Tseng, Li-Hong; Gil, Ana M

    2008-08-01

    The metabolic profiling of human amniotic fluid (HAF) is of potential interest for the diagnosis of disorders in the mother or the fetus. In order to build a comprehensive metabolite database for HAF, hyphenated NMR has been used, for the first time, for systematic HAF profiling. Experiments were carried out using reverse-phase (RP) and ion-exchange liquid chromatography (LC), in order to detect less and more polar compounds, respectively. RP-LC conditions achieved good separation of amino acids, some sugars, and xanthines. Subsequent NMR and MS analysis enabled the rapid identification of 30 compounds, including 3-methyl-2-oxovalerate and 4-aminohippurate identified in HAF for the first time, to our knowledge. Under ion-exchange LC conditions, a different set of 30 compounds was detected, including sugars, organic acids, several derivatives of organic acids, and amino acids. In this experiment, five compounds were identified for the first time in HAF: D-xylitol, amino acid derivatives (N-acetylalanine, N-acetylglycine, 2-oxoleucine), and isovalerate. The nonendogenous nature of some metabolites (caffeine, paraxanthine, D-xylitol, sorbitol) is discussed. Hyphenated NMR has allowed the rapid detection of approximately 60 metabolites in HAF, some of which are not detectable by standard NMR due to low abundance (microM) and signal overlap thus enabling an extended metabolite database to be built for HAF.

  4. Comparative proteomic analysis of human amniotic fluid supernatants with Down syndrome using mass spectrometry.

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    Park, Jisook; Cha, Dong-Hyun; Jung, Jin Woo; Kim, Young Hwan; Lee, Sook Hwan; Kim, Youngjun; Kim, Kwang Pyo

    2010-06-01

    Down syndrome (DS) is an abnormality of the 21st chromosome that commonly occurs in children born to advanced age women. Amniotic fluid (AF) is usually collected from such women for prenatal diagnosis. This study analyzed human AF supernatants (AFS) by mass spectrometric (MS) approach to search for candidate biomarkers of DS pregnancy. AFS were collected from advanced age pregnant women at 16-18th weeks of gestation by amniocentesis for cytogenetic analysis. AFS from pregnancies carrying DS (n=4) or chromosomally normal (n=6) fetuses, as revealed by cytogenetic analysis, were then subjected to global protein profiling by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Affinity chromatography was applied prior to LC-ESI-MS/MS to minimize the masking effect of highly abundant albumin and immunoglobulin and thereby, increased the diversity of identified proteins. Hereby, at least 30 AFS proteins were newly identified and 44 AFS proteins were found to be differentially expressed between DS and normal cases. Six of these proteins were unique to DS cases while 11 proteins were unique to chromosomally normal cases. In addition, 19 AFS proteins were down-regulated and 8 were up-regulated in DS cases with varying fold changes. A western blot analysis confirmed the LC-ESI-MS/MS data that combined detection of Apolipoprotein A-II (apo A-II) and alpha-fetoprotein (AFP) could be a potential tool for diagnosing DS cases.

  5. Potential of NMR spectroscopy for the study of human amniotic fluid.

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    Graça, Gonçalo; Duarte, Iola F; Goodfellow, Brian J; Barros, António S; Carreira, Isabel M; Couceiro, Ana Bela; Spraul, Manfred; Gil, Ana M

    2007-11-01

    1D and 2D 800 MHz high-resolution nuclear magnetic resonance spectroscopy of human amniotic fluid (HAF) enabled the identification of approximately 50 metabolites. In addition, liquid chromatography-NMR and diffusion ordered spectroscopy (DOSY) allowed signal overlap to be reduced and the characterization of higher molecular weight (Mw) components, respectively. Indeed, the DOSY spectrum of a Mw >10 kDa HAF fraction enabled three protein families, differing in average Mw, to be detected and may therefore be of potential value in the study of disorder-related variations in HAF protein profiles. The effects of freeze-drying, storage at -20 or -70 degrees C, and freeze-thawing cycles on HAF compositional stability were investigated, as well as stability at room temperature (to account for overnight data acquisition runs). These data are the basis for establishing statistically validated correlations between HAF NMR data and any physiological disorders of the fetus/mother. Freeze-drying caused signal loss for urea, ethanol, and compounds resonating at 2.22 and 1.17 ppm. Storage at -70 degrees C or lower is recommended since only minor compositional changes were observed, affecting mainly acetate and pyruvate. Freeze-thaw cycles did not cause significant compositional changes, and room-temperature stability studies indicated a 4-5 h maximum period of handling/acquisition time to ensure HAF stability.

  6. Comparisons of human amniotic mesenchymal stem cell viability in FDA-approved collagen-based scaffolds: Implications for engineered diaphragmatic replacement.

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    Shieh, Hester F; Graham, Christopher D; Brazzo, Joseph A; Zurakowski, David; Fauza, Dario O

    2017-06-01

    We sought to examine amniotic fluid mesenchymal stem cell (afMSC) viability within two FDA-approved collagen-based scaffolds, as a prerequisite to clinical translation of afMSC-based engineered diaphragmatic repair. Human afMSCs were seeded in a human-derived collagen hydrogel and in a bovine-derived collagen sheet at 3 matching densities. Cell viability was analyzed at 1, 3, and 5days using an ATP-based 3D bioluminescence assay. Statistical comparisons were by ANOVA (Pcollagen hydrogel when compared with a collagen sheet. Cell viability can be further optimized by seeding density and time in 3D culture. These data further support the regulatory viability of clinical trials of engineered diaphragmatic repair. N/A (animal and laboratory study). Copyright © 2017 Elsevier Inc. All rights reserved.

  7. An epidermal stem cells niche microenvironment created by engineered human amniotic membrane.

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    Ji, Shi-zhao; Xiao, Shi-chu; Luo, Peng-fei; Huang, Guo-feng; Wang, Guang-yi; Zhu, Shi-hui; Wu, Min-juan; Xia, Zhao-fan

    2011-11-01

    How to amplify epidermal stem cells (ESCs) rapidly is a challenging crux in skin tissue engineering research. The present study describes the preparation of 3D micronized (300-600 μm) amniotic membrane (mAM) by means of repeated freeze-thawing cycles to deplete cell components and homogenized with a macrohomogenizer in liquid nitrogen. This newly prepared mAM not only possessed the characteristics of a microcarrier but completely retained the basement membrane structure and abundant active substances such as NGF, HGF, KGF, bFGF, TGF-β1 and EGF in the AM matrix. The result showed that mAM combined with rotary cell culture system (RCCS) was able to amplify ESCs quickly. The relative cell viability at day 7 and 14 was significantly higher than that of the conventional 2D plate culture (326 ± 28% and 535 ± 47% versus 232 ± 21% and 307 ± 32%, P human body favorable for ex vivo culture and amplification of ESCs but can be used as the dermal scaffold in constructing a skin substitute containing ESCs for the repair of full-thickness skin defects. Copyright © 2011 Elsevier Ltd. All rights reserved.

  8. Amniotic fluid "sludge"; prevalence and clinical significance of it in asymptomatic patients at high risk for spontaneous preterm delivery.

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    Adanir, Ilknur; Ozyuncu, Ozgur; Gokmen Karasu, Ayse Filiz; Onderoglu, Lutfu S

    2018-01-01

    The aim of our study is to determine prevalence and clinical significance of the presence of amniotic fluid "sludge" among asymptomatic patients at high-risk for spontaneous preterm delivery, prospectively. In our study, 99 patients at high risk for spontaneous preterm delivery were evaluated for the presence of amniotic fluid sludge with transvaginal ultrasonography at 20-22, 26-28, and 32-34 gestational weeks, prospectively; between August 2009 and October 2010 in Hacettepe University Hospital. And, these patients were followed up for their delivery weeks and pregnancy outcomes. We defined the high-risk group as the patients possessing one or more of the followings; a history of spontaneous preterm delivery, recent urinary tract infections, polyhydramnios, uterine leiomyomas, müllerian duct anomalies, and history of cone biyopsy or LEEP. Patients with multiple gestations, placenta previa, fetal anomalies, or symptoms of preterm labor at first examination were excluded. We have obtained ethical board approval from Hacettepe University (16.07.2009-HEK/No:09-141-59). The prevalence of amniotic fluid sludge in the study population was 19,6% (18/92). The rates of spontaneous preterm delivery at preterm delivery (p = 0.002). A higher proportion of neonates born to patients with amniotic fluid sludge had a neonatal morbidity (50% (9/18) vs. 24,3% (18/74), p = 0.044) and died in the perinatal period, (p = 0,013) than those born to patients without sludge. When we combined sludge and cervical lenght (CL) (preterm delivery; it catched more women with preterm delivery, (p = 0.000). While sensitivity of sludge was 37,5%, and sensitivity of CL was 34%, sensitivity of "sludge positive or CL ≤25 mm" was 56% for preterm birth (PTB) in high-risk group. The prevalence of amniotic fluid sludge is 19,6% and "sludge" is an independent risk factor for spontaneous preterm delivery among asymptomatic patients at high-risk for spontaneous preterm delivery. PTB is

  9. Second-trimester amniotic fluid corticotropin-releasing hormone and urocortin in relation to maternal stress and fetal growth in human pregnancy.

    Science.gov (United States)

    La Marca-Ghaemmaghami, Pearl; Dainese, Sara M; Stalla, Günter; Haller, Marina; Zimmermann, Roland; Ehlert, Ulrike

    2017-05-01

    This study explored the association between the acute psychobiological stress response, chronic social overload and amniotic fluid corticotropin-releasing hormone (CRH) and urocortin (UCN) in 34 healthy, second-trimester pregnant women undergoing amniocentesis. The study further examined the predictive value of second-trimester amniotic fluid CRH and UCN for fetal growth and neonatal birth outcome. The amniocentesis served as a naturalistic stressor, during which maternal state anxiety and salivary cortisol was measured repeatedly and an aliquot of amniotic fluid was collected. The pregnant women additionally completed a questionnaire on chronic social overload. Fetal growth parameters were obtained at amniocentesis using fetal ultrasound biometry and at birth from medical records. The statistical analyzes revealed that the acute maternal psychobiological stress response was unassociated with the amniotic fluid peptides, but that maternal chronic overload and amniotic CRH were positively correlated. Moreover, amniotic CRH was negatively associated with fetal size at amniocentesis and positively with growth in size from amniocentesis to birth. Hardly any studies have previously explored whether acute maternal psychological stress influences fetoplacental CRH or UCN levels significantly. Our findings suggest that (i) chronic, but not acute maternal stress may affect fetoplacental CRH secretion and that (ii) CRH is complexly involved in fetal growth processes as previously shown in animals.

  10. Therapeutic efficacy of amniotic membrane stem cells and adipose tissue stem cells in rats with chemically induced ovarian failure

    Science.gov (United States)

    Fouad, Hanan; Sabry, Dina; Elsetohy, Khaled; Fathy, Naglaa

    2015-01-01

    The present study was conducted to compare between the therapeutic efficacies of human amniotic membrane-derived stem cells (hAM-MSCs) vs. adipose tissue derived stem cells (AD-MSCs) in cyclophosphamide (CTX)-induced ovarian failure in rats. Forty-eight adult female rats were included in the study; 10 rats were used as control group. Thirty-eight rats were injected with CTX to induce ovarian failure and divided into four groups: ovarian failure (IOF) (IOF group), IOF + phosphate buffer saline (PBS group), IOF + hAM-MSCs group and IOF + AD-MSCs group. Serum levels of FSH and estradiol (E2) were assessed. Histopathological examination of the ovarian tissues was performed and quantitative gene expressions of Oct-4, Stra8 and integrin beta-1 genes were conducted by quantitative real time PCR. Results showed that IOF and IOF + PBS rat groups exhibited decreased ovarian follicles, increased interstitial fibrosis with significant decrease of serum E2, significant increase serum FSH level and significant down-regulation of Stra8 and integrin beta-1. In hAM-MSCs and AD-MSCs rat groups, there were increased follicles and corpora with evident the presence of oocytes, significant increase in serum E2, significant decrease in serum FSH levels (in hAM-MSCs treated group only) and significant up-regulation of the three studied genes with higher levels in hAM-MSCs treated rats group when compared to AD-MSCs treated rats group. In Conclusion, administration of either hAM-derived MSCs or AD-MSCs exerts a significant therapeutic efficacy in chemotherapy induced ovarian insult in rats. hAM-MSCs exert higher therapeutic efficacy as compared to AD-MSCs. PMID:26966564

  11. Activation of mitosis and angiogenesis in diabetes-impaired wound healing by processed human amniotic fluid.

    Science.gov (United States)

    Bazrafshan, Ameneh; Owji, Mohammad; Yazdani, Maryam; Varedi, Masoumeh

    2014-05-15

    Functional characterization of human amniotic fluid (AF) proteome, 845 proteins, has revealed that top three functions are cell proliferation, movement and differentiation, events fundamental to development, and tissue repair. Although these findings fortify the idea that AF components play roles in regeneration-like fetal wound healing, it is not known whether the components endure processing. Therefore, we processed AF and tested its effects on diabetes-impaired wound healing in an animal model. Through a germfree procedure, mature and premature AF samples were collected, respectively, from the mothers of full-term and preterm infants. Excisional wounds were generated on the dorsum of diabetic rats. Wounds were treated on day 3 and harvested on day 7 postwounding. Proliferating cell nuclear antigen and alpha-smooth muscles actin, markers for mitosis and angiogenesis, respectively, were assessed by in situ immunodetection method. Significant increases in the rate of wound closure and proliferating cell nuclear antigen-expressing cells were observed in AF-treated wounds when compared with that of sham and control wounds. Likewise, the number of large vessels was significantly increased in the wounds treated with the AF. However, population of myofibroblasts was not affected by the treatment. The mature and premature AF were almost equally effective. Our data, for the first time, show that processed AF accelerates diabetes-impaired wound healing by activating mitosis and angiogenesis, indicating that bioactive molecules in AF may endure processing. We believe that processed forms of this naturally designed "Cocktail" of bioactive molecules may have multiple clinical applications. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. Enhanced generation of retinal progenitor cells from human retinal pigment epithelial cells induced by amniotic fluid

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    Sanie-Jahromi Fatemeh

    2012-04-01

    Full Text Available Abstract Background Retinal progenitor cells are a convenient source of cell replacement therapy in retinal degenerative disorders. The purpose of this study was to evaluate the expression patterns of the homeobox genes PAX6 and CHX10 (retinal progenitor markers during treatment of human retinal pigment epithelium (RPE cells with amniotic fluid (AF, RPE cells harvested from neonatal cadaver globes were cultured in a mixture of DMEM and Ham's F12 supplemented with 10% FBS. At different passages, cells were trypsinized and co-cultured with 30% AF obtained from normal fetuses of 1416 weeks gestational age. Results Compared to FBS-treated controls, AF-treated cultures exhibited special morphological changes in culture, including appearance of spheroid colonies, improved initial cell adhesion and ordered cell alignment. Cell proliferation assays indicated a remarkable increase in the proliferation rate of RPE cells cultivated in 30% AF-supplemented medium, compared with those grown in the absence of AF. Immunocytochemical analyses exhibited nuclear localization of retinal progenitor markers at a ratio of 33% and 27% for CHX10 and PAX6, respectively. This indicated a 3-fold increase in retinal progenitor markers in AF-treated cultures compared to FBS-treated controls. Real-time PCR data of retinal progenitor genes (PAX6, CHX10 and VSX-1 confirmed these results and demonstrated AF's capacity for promoting retinal progenitor cell generation. Conclusion Taken together, the results suggest that AF significantly promotes the rate of retinal progenitor cell generation, indicating that AF can be used as an enriched supplement for serum-free media used for the in vitro propagation of human progenitor cells.

  13. Meconium-stained amniotic fluid and meconium aspiration syndrome: a prospective study.

    Science.gov (United States)

    Bhat, R Y; Rao, A

    2008-09-01

    The incidence of meconium aspiration syndrome (MAS), associated perinatal factors, morbidity and deaths varies widely. This study aimed to assess the perinatal attributes and morbidity associated with MAS. Over a 2-year period, all neonates born through meconium-stained amniotic fluid (MSAF) were observed for respiratory distress (RD). Birth details, chest radiograph (CXR) and clinical course were documented. Neonates with consistent CXR findings whose RD could not otherwise be explained were defined as MAS. Of 409 neonates born through MSAF, meconium was thick in 196 (47.9%). Fifty-five (13.4%) had RD and 45 (11.3%) were consistent with MAS. Six (1.5%) neonates died. Mean (SD) birthweight and gestation of MAS infants were 2721.9 (510.2) g and 38.67 (1.09) weeks, respectively. About one-third were of low birthweight and 28 were born by caesarean section. On univariate analysis, caesarean delivery, meconium in the trachea and thick meconium were the significant perinatal factors for the development of MAS. On multiple regression analysis, thick meconium was the only independent factor for MAS (OR 7.08, 95% CI 3.08-16.27, psyndrome was observed in 12 (26.7%), persistent pulmonary hypertension in seven and severe asphyxia and late sepsis in two each. Thirteen (28.9%) required mechanical ventilation and six (13.3%) died. MAS occurred in 11.3% of neonates born through MSAF. Thick meconium was found to be an important causative factor. Nearly one-third of infants with MAS required mechanical ventilation and 13.3% died.

  14. Amniotic liquid derived stem cells as reservoir of secreted angiogenic factors capable of stimulating neo-arteriogenesis in an ischemic model.

    Science.gov (United States)

    Mirabella, Teodelinda; Teodelinda, Mirabella; Cilli, Michele; Michele, Cilli; Carlone, Sebastiano; Sebastiano, Carlone; Cancedda, Ranieri; Ranieri, Cancedda; Gentili, Chiara; Chiara, Gentili

    2011-05-01

    Most urgent health problems are related to a blood vessel formation failure. The use of stem cells from different sources or species for both in vitro and in vivo engineering of endothelium does not necessarily imply their direct commitment towards a vascular phenotype. In the present study, we used human amniotic fluid stem cells (AFSC) to evoke a strong angiogenic response in murine recipients, in terms of host guided-regeneration of new vessels, and we demonstrated that the AFSC secretome is responsible for the vascularising properties of these cells. We indentified in AFSC conditioned media (ACM) pro-angiogenic soluble factors, such as MCP-1, IL-8, SDF-1, VEGF. Our in vitro results suggest that ACM are cytoprotective, pro-differentiative and chemoattractive for endothelial cells. We also tested ACM on a pre-clinical model of hind-limb ischemic mouse, concluding that ACM contain mediators that promote the neo-arteriogenesis, as remodelling of pre-existing collateral arteries to conductance vessels, thus preventing the capillary loss and the tissue necrosis of distal muscles. In line with the current regenerative medicine trend, in the present study we assert the concept that stem cell-secreted mediators can guide the tissue repair by stimulating or recruiting host reparative cells. Copyright © 2011 Elsevier Ltd. All rights reserved.

  15. Human amniotic epithelial cells as feeder layer to derive and maintain human embryonic stem cells from poor-quality embryos

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    Daniela Ávila-González

    2015-09-01

    Full Text Available Data from the literature suggest that human embryonic stem cell (hESC lines used in research do not genetically represent all human populations. The derivation of hESC through conventional methods involve the destruction of viable human embryos, as well the use of mouse embryonic fibroblasts as a feeder layer, which has several drawbacks. We obtained the hESC line (Amicqui-1 from poor-quality (PQ embryos derived and maintained on human amniotic epithelial cells (hAEC. This line displays a battery of markers of pluripotency and we demonstrated the capacity of these cells to produce derivates of the three germ layers.

  16. Evaluation of two commercially available ELISA kits for the determination of melatonin concentrations in amniotic fluid throughout pregnancy.

    Science.gov (United States)

    Bagci, Soyhan; Altuntas, Özlem; Katzer, David; Berg, Christoph; Willruth, Arne; Reutter, Heiko; Bartmann, Peter; Müller, Andreas; Zur, Berndt

    2017-01-01

    Background The aim of the present study is to evaluate the utility of extraction versus non-extraction-based commercial melatonin ELISA kits for determining the melatonin concentration in amniotic fluid obtained in early and late pregnancy. Methods Pregnancy duration less than 28 weeks was defined as early and from 28 weeks until delivery as late gestation. Nine samples were obtained in early and 18 in late pregnancy. Two commercially available melatonin ELISA kits (melatonin ELISA RE54021, including methanol-based extraction and direct saliva melatonin ELISA RE 54041, not including an extraction step, both from IBL-International, Germany) were used to determine melatonin concentrations in amniotic fluid. Results The mean melatonin concentration in ELISAs assayed by the non-extraction was significantly lower than those assayed after extraction. Subgroup analysis showed that there was no significant difference between melatonin concentration measured by non-extraction versus extraction ELISA in early pregnancy (11.2 ± 7.4 vs. 12.2 ± 7.7, respectively, P = 0.463) but that the mean melatonin concentration in late pregnancy was significantly lower when assayed by non-extraction ELISA than when assayed by extraction ELISA (14.8 ± 9.3 vs. 145.1 ± 179.3, respectively; P pregnancy was rather poor (r 2  = 0.271, P = 0.022), as opposed to the good correlation found in early pregnancy (r 2  = 0.929, P melatonin assay without an extraction step, such as direct saliva ELISA, does not seem to be a valid method to determine the melatonin concentration of amniotic fluid, especially in late gestation.

  17. Efficacy of betamethasone on the fetal motion and biophysical profile and amniotic fluid index in preterm fetuses.

    Science.gov (United States)

    Abbasalizadeh, Shamsi; Pharabar, Zahra Neghadan; Abbasalizadeh, Fatmeh; Ghojazadeh, Morteza; Goldust, Mohamad

    2013-11-15

    The term ofpreterm birth is used to define the premature neonates considering pregnancy age. In less than 34 week pregnancies, corticosteroids are prescribed to promote embryos' lung maturity. The presents study aimed at evaluating effects of betamethasone injection on feeling embryo motion by mother and index and biophysical profile in preterm pregnancies. In a descriptive-analytical study, 40 pregnant women with the pregnancy age of 30-34 weeks were evaluated. Embryo motion and index and biophysical profile of the amniotic fluid were checked before prescription of double dosage of muscular betamethasone (12 mg) at a 24 h time interval. The injection was repeated for 24 and 48 h after the first injection. The resulted outcomes were compared with those results related to before betamethasone injection. In this study, there was statistically meaningful relationship between embryo motions before injection of betamethasone and 12 h after its injection (p = 0.03). Also, there was a significant relationship between embryo motions 24 and 48 h after injection of betamethasone (p = 0.001). In other words, the embryo motions decreased 12 h after injection of betamethasone. They were improved 48 h after betamethasone injection. But, index and biophysical profile results of amniotic fluid were left unchanged. Application of betamethasone leads to evident but transient decrease in embryo motions. Although motion element of index and biophysical profile of amniotic fluid which is one of the tests used in evaluating the embryo health is fixed and normal, it can be concluded that injection of betamethasone may not affect embryo health.

  18. Detection and sequencing of Zika virus from amniotic fluid of fetuses with microcephaly in Brazil: a case study.

    Science.gov (United States)

    Calvet, Guilherme; Aguiar, Renato S; Melo, Adriana S O; Sampaio, Simone A; de Filippis, Ivano; Fabri, Allison; Araujo, Eliane S M; de Sequeira, Patricia C; de Mendonça, Marcos C L; de Oliveira, Louisi; Tschoeke, Diogo A; Schrago, Carlos G; Thompson, Fabiano L; Brasil, Patricia; Dos Santos, Flavia B; Nogueira, Rita M R; Tanuri, Amilcar; de Filippis, Ana M B

    2016-06-01

    The incidence of microcephaly in Brazil in 2015 was 20 times higher than in previous years. Congenital microcephaly is associated with genetic factors and several causative agents. Epidemiological data suggest that microcephaly cases in Brazil might be associated with the introduction of Zika virus. We aimed to detect and sequence the Zika virus genome in amniotic fluid samples of two pregnant women in Brazil whose fetuses were diagnosed with microcephaly. In this case study, amniotic fluid samples from two pregnant women from the state of Paraíba in Brazil whose fetuses had been diagnosed with microcephaly were obtained, on the recommendation of the Brazilian health authorities, by ultrasound-guided transabdominal amniocentesis at 28 weeks' gestation. The women had presented at 18 weeks' and 10 weeks' gestation, respectively, with clinical manifestations that could have been symptoms of Zika virus infection, including fever, myalgia, and rash. After the amniotic fluid samples were centrifuged, DNA and RNA were extracted from the purified virus particles before the viral genome was identified by quantitative reverse transcription PCR and viral metagenomic next-generation sequencing. Phylogenetic reconstruction and investigation of recombination events were done by comparing the Brazilian Zika virus genome with sequences from other Zika strains and from flaviviruses that occur in similar regions in Brazil. We detected the Zika virus genome in the amniotic fluid of both pregnant women. The virus was not detected in their urine or serum. Tests for dengue virus, chikungunya virus, Toxoplasma gondii, rubella virus, cytomegalovirus, herpes simplex virus, HIV, Treponema pallidum, and parvovirus B19 were all negative. After sequencing of the complete genome of the Brazilian Zika virus isolated from patient 1, phylogenetic analyses showed that the virus shares 97-100% of its genomic identity with lineages isolated during an outbreak in French Polynesia in 2013, and that in

  19. Amniotic fluid cathelicidin in PPROM pregnancies: from proteomic discovery to assessing its potential in inflammatory complications diagnosis.

    Science.gov (United States)

    Tambor, Vojtech; Kacerovsky, Marian; Andrys, Ctirad; Musilova, Ivana; Hornychova, Helena; Pliskova, Lenka; Link, Marek; Stulik, Jiri; Lenco, Juraj

    2012-01-01

    Preterm prelabor rupture of membranes (PPROM) complicated by microbial invasion of the amniotic cavity (MIAC) leading to histological chorioamnionitis (HCA) significantly impacts perinatal morbidity. Unfortunately, no well-established tool for identifying PPROM patients threatened by these disorders is available. We performed an unbiased exploratory analysis of amniotic fluid proteome changes due to MIAC and HCA. From among the top five proteins that showed the most profound and significant change, we sought to confirm results concerning cathelicidin (P49913, CAMP_HUMAN), since an ELISA kit was readily available for this protein. In our exploratory proteomic study, cathelicidin showed a ∼6-fold higher concentration in PPROM patients with confirmed MIAC and HCA. We verified significantly higher levels of cathelicidin in exploratory samples (women without both MIAC and HCA: median 1.4 ng/ml; women with both conditions confirmed: median 3.6 ng/ml; p = 0.0003). A prospective replication cohort was used for independent validation and for assessment of cathelicidin potential to stratify women with MIAC leading to HCA from women in whom at least one of these conditions was ruled out. We confirmed the association of higher amniotic fluid cathelicidin levels with MIAC leading to HCA (the presence of both MIAC and HCA: median 3.1 ng/ml; other women: median 1.4 ng/ml; p<0.0001). A cathelicidin concentration of 4.0 ng/ml was found to be the best cut-off point for identifying PPROM women with both MIAC and HCA. When tested on the validation cohort, a sensitivity of 48%, a specificity of 90%, a likelihood ratio of 5.0, and an area under receiver-operating characteristic curve of 71% were achieved for identification of women with MIAC leading to HCA. Our multi-stage study suggests cathelicidin as a candidate marker that should be considered for a panel of amniotic fluid proteins permitting identification of PPROM women with MIAC leading to HCA.

  20. Amniotic fluid cathelicidin in PPROM pregnancies: from proteomic discovery to assessing its potential in inflammatory complications diagnosis.

    Directory of Open Access Journals (Sweden)

    Vojtech Tambor

    Full Text Available BACKGROUND: Preterm prelabor rupture of membranes (PPROM complicated by microbial invasion of the amniotic cavity (MIAC leading to histological chorioamnionitis (HCA significantly impacts perinatal morbidity. Unfortunately, no well-established tool for identifying PPROM patients threatened by these disorders is available. METHODOLOGY/PRINCIPAL FINDINGS: We performed an unbiased exploratory analysis of amniotic fluid proteome changes due to MIAC and HCA. From among the top five proteins that showed the most profound and significant change, we sought to confirm results concerning cathelicidin (P49913, CAMP_HUMAN, since an ELISA kit was readily available for this protein. In our exploratory proteomic study, cathelicidin showed a ∼6-fold higher concentration in PPROM patients with confirmed MIAC and HCA. We verified significantly higher levels of cathelicidin in exploratory samples (women without both MIAC and HCA: median 1.4 ng/ml; women with both conditions confirmed: median 3.6 ng/ml; p = 0.0003. A prospective replication cohort was used for independent validation and for assessment of cathelicidin potential to stratify women with MIAC leading to HCA from women in whom at least one of these conditions was ruled out. We confirmed the association of higher amniotic fluid cathelicidin levels with MIAC leading to HCA (the presence of both MIAC and HCA: median 3.1 ng/ml; other women: median 1.4 ng/ml; p<0.0001. A cathelicidin concentration of 4.0 ng/ml was found to be the best cut-off point for identifying PPROM women with both MIAC and HCA. When tested on the validation cohort, a sensitivity of 48%, a specificity of 90%, a likelihood ratio of 5.0, and an area under receiver-operating characteristic curve of 71% were achieved for identification of women with MIAC leading to HCA. CONCLUSIONS: Our multi-stage study suggests cathelicidin as a candidate marker that should be considered for a panel of amniotic fluid proteins permitting identification

  1. Amniotic fluid embolism incidence, risk factors and outcomes: a review and recommendations

    Directory of Open Access Journals (Sweden)

    Knight Marian

    2012-02-01

    Full Text Available Abstract Background Amniotic fluid embolism (AFE is a rare but severe complication of pregnancy. A recent systematic review highlighted apparent differences in the incidence, with studies estimating the incidence of AFE to be more than three times higher in North America than Europe. The aim of this study was to examine population-based regional or national data from five high-resource countries in order to investigate incidence, risk factors and outcomes of AFE and to investigate whether any variation identified could be ascribed to methodological differences between the studies. Methods We reviewed available data sources on the incidence of AFE in Australia, Canada, the Netherlands, the United Kingdom and the USA. Where information was available, the risk factors and outcomes of AFE were examined. Results The reported incidence of AFE ranged from 1.9 cases per 100 000 maternities (UK to 6.1 per 100 000 maternities (Australia. There was a clear distinction between rates estimated using different methodologies. The lowest estimated incidence rates were obtained through validated case identification (range 1.9-2.5 cases per 100 000 maternities; rates obtained from retrospective analysis of population discharge databases were significantly higher (range 5.5-6.1 per 100 000 admissions with delivery diagnosis. Older maternal age and induction of labour were consistently associated with AFE. Conclusions Recommendation 1: Comparisons of AFE incidence estimates should be restricted to studies using similar methodology. The recommended approaches would be either population-based database studies using additional criteria to exclude false positive cases, or tailored data collection using existing specific population-based systems. Recommendation 2: Comparisons of AFE incidence between and within countries would be facilitated by development of an agreed case definition and an agreed set of criteria to minimise inclusion of false positive cases for

  2. Association of Parasite Load Levels in Amniotic Fluid With Clinical Outcome in Congenital Toxoplasmosis.

    Science.gov (United States)

    Yamamoto, Lidia; Targa, Lília S; Sumita, Laura M; Shimokawa, Paulo T; Rodrigues, Jonatas C; Kanunfre, Kelly A; Okay, Thelma S

    2017-08-01

    To correlate neonatal and infant clinical outcome with parasite load in amniotic fluid (AF). We conducted a retrospective cohort study of 122 children whose mothers had toxoplasmosis during pregnancy. The children were monitored from birth to 12 months old. Stored AF samples were obtained at maternal diagnosis and tested by quantitative polymerase chain reaction. Gestational age at maternal infection, quantitative polymerase chain reaction results, neonatal anti-Toxoplasma gondii immunoglobulin (Ig) M, and clinical outcome at 12 months were correlated. Maternal infection occurred in 18 of 122 (14.7%) and 104 of 122 (85.2%) women in the first and second trimesters, respectively. At birth, IgM was present in 107 of 122 (87.7%) neonates and 36 (29.5%) were symptomatic. Of these, half occurred in the first and the other half in the second trimester and 6 of 36 had severe infections (16.7% of symptomatic, 4.9% of total), all infected in the first trimester. Parasite load levels were highly variable (median 35 parasites/mL, range 2-30,473). Logistic regression correlated symptomatic infection with gestational age (odds ratio [OR] 0.47, CI 0.31-0.73) and parasite load (OR 2.04, CI 1.23-3.37), but not with positive IgM (OR 6.81, CI 0.86-53.9). Negative correlations were found between gestational age and parasite load (rs -0.780, CI -0.843 to -0.696), gestational age and symptoms (rs -0.664, CI -0.755 to -0.547), but not gestational age and IgM (rs -0.136, CI -0.311 to 0.048). Parasite load levels distributed by percentile showed that all symptomatic patients appeared from the 75th percentile and all severe infections from the 95th percentile. Load rankings showed doubled the OR for each 20 parasite/mL increment. Parasite load was associated with symptomatic infections (area under the curve 0.959, CI 0.908-0.987) as well as gestational age (area under the curve 0.918, CI 0.855-0.960) and both parameters combined (area under the curve 0.969, CI 0.920-0.992). Parasite load in

  3. Detection and Comparison of Cytomegalovirus DNA Levels in Amniotic Fluid and Fetal Ascites in a Second-Trimester Fetus With Massive Ascites, Hyperechogenic Bowel, Ventriculomegaly and Intrauterine Growth Restriction

    Directory of Open Access Journals (Sweden)

    Chih-Ping Chen

    2010-06-01

    Conclusion: A prenatal diagnosis of fetal ascites in association with ventriculomegaly, hyperechogenic bowel and intrauterine growth restriction should alert physicians to congenital CMV infection in addition to aneuploidy. The present case provides evidence that CMV DNA levels are higher in amniotic fluid (amniocytes and amniotic fluid supernatant than in ascites (ascitic cells and ascitic supernatant in cases of congenital CMV infection.

  4. Transplantation with cultured stem cells derived from the human amniotic membrane for corneal alkali burns: an experimental study.

    Science.gov (United States)

    Zeng, Wei; Li, Yanwei; Zeng, Guangwei; Yang, Bo; Zhu, Yu

    2014-01-01

    Amniotic membranes (AM) have been used in a wide range of clinical applications. We successfully extracted mesenchymal stem cells (MSCs) from human AM, but little is known about the use and efficacy of human amniotic membrane-derived mesenchymal stem cells (hAM-dMSCs) for the treatment of alkali burns. We utilized hAM-dMSCs transplantation, AM grafting, and their combined use in the treatment of alkali burns. An experimental model in rabbits was devised to analyze the use of these techniques with immunocytochemistry and ELISA. The survival and migration of hAM-dMSCs labeled by SPION in the host were assessed with Prussian blue staining. Compared with the control group, the treated groups demonstrated faster reconstruction of the corneal epithelium, and lower levels of corneal opacification and neovascularization within corneal alkali burns. Furthermore, dark blue-stained particles were detected in the limbus corneae at day 28. These results demonstrated the ability of hAM-dMSCs to enhance epithelial healing and reduce corneal opacification and neovascularization in corneal alkali wounds.

  5. Survey of Pregnancy Outcome in Preterm Premature Rupture of Membranes with Amniotic Fluid Index <5 and ≥5

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    Fatemeh Tavassoli

    2010-04-01

    Full Text Available Objectives: Preterm premature rupture of membranes (PPROM is among the most important causes of perinatal morbidity and mortality. The aim of this study is to survey the pregnancy outcomes in preterm premature rupture of membranes with an amniotic fluid index of 5.Methods: This prospective cohort study was performed on 137 pregnant women complicated by preterm premature rupture of membranes (PPROM with a gestational age of 28-34 weeks during October 2006 to October 2008. The patients were divided in two groups according to their amniotic fluid index; AFI<5 (77cases, AFI≥5 (60cases. The Chi-squared test for qualitative variables and T-student test for quantitative variables were used to analyze the results.Results: The results showed that there was no significant difference in terms of the number of pregnancies, gestational age at rupture of membranes and birthweight between the two groups. However, the results demonstrated that the patients with AFI<5 exhibited a significantly shorter latency period (p=0.049, a higher rate of cesarean due to fetal distress (p=0.008, a lower neonatal Apgar score in the first minute (p=0.0127 and a higher rate of neonatal death during the first week (p=0.045.Conclusion: Overall, PPROM with oligohydroamnios is associated with shorter latency, higher rate of C/S, higher rate of early neonatal death and lower neonatal Apgar.

  6. Comparative Study of Pregnancy Outcome in Premature Rupture of Membranes With Amniotic Fluid Indices of less and more than 5

    Directory of Open Access Journals (Sweden)

    F Tavassoli

    2010-03-01

    Full Text Available Introduction: Preterm premature rupture of the membranes (PPROM is one of the most important causes of perinatal morbidity and mortality. The aim of this study was to evaluate pregnancy outcome in preterm premature rupture of the membranes with amniotic fluid index less than 5 and more than 5. Methods: This prospective study was performed on 137 pregnant women with gestational age of 28-34 weeks with PPROM from Oct 2006 to Oct 2008. The patients were divided in two groups according to amniotic fluid index: AFI<5 (77 cases, AFI≥5 (60 cases. Chi-square test was used for qualitative variables, while T-student test was used for quantitative test. Results: Both groups were similar with respect to number of pregnancies, gestational age at rupture of the membranes and birth weight. In the group with AFI<5, applied latency was significantly shorter (PV=0.049, rate of cesarean section was higher due to fetal distress (PV=0.008, neonatal Apgar score in first minute was lower during the first week (PV=0.0127 and the rate of neonatal death was higher (PV=0.045 during the first week. Conclusion: An AFI<5 cm after PPROM is associated with earlier delivery, higher rate of cesarean due to fetal distress, higher rate of neonatal death, and lower neonatal Apgar score in first minute during the first week

  7. Windowed direct exponential curve resolution quantification of nuclear magnetic resonance spectroscopy with applications to amniotic fluid metabonomics

    International Nuclear Information System (INIS)

    Botros, L.L.

    2007-01-01

    This thesis presents a quantitative protocol of proton nuclear magnetic resonance ( 1 H NMR) that allows the determination of human amniotic fluid metabolite concentrations, which are then used in a metabonomic study to establish patient health during gestation. 1 H NMR free inductive decays (FIDs) of 258 human amniotic fluid samples from a 500MHz spectrometer are acquired. Quantitative analyses methods in both the frequency- and time-domain are carried out and compared. Frequency-domain analysis is accomplished by integration of the metabolite peaks before and after the inclusion of a known standard addition of alanine. Time-domain analysis is accomplished by the direct exponential curve resolution algorithm (DECRA). Both techniques are assessed by applications to calibration biological solutions and a simulated data set. The DECRA method proves to be a more accurate and precise route for quantitative analysis, and is included in the developed protocol. Well-defined peaks of various components are visible in the frequency-domain 1 H NMR spectra, including lactate, alanine, acetate, citrate, choline, glycine, and glucose. All are quantified with the proposed protocol. Statistical t-test and notched box and whisker plots are used to compare means of metabolite concentrations for diabetic and normal patients. Glucose, glycine, and choline are all found to correlate with gestational diabetes mellitus early in gestation. With further development, time-domain quantitative 1 H NMR has potential to become a robust diagnostic tool for gestational health. (author)

  8. Windowed direct exponential curve resolution quantification of nuclear magnetic resonance spectroscopy with applications to amniotic fluid metabonomics

    Energy Technology Data Exchange (ETDEWEB)

    Botros, L.L

    2007-07-01

    This thesis presents a quantitative protocol of proton nuclear magnetic resonance ({sup 1}H NMR) that allows the determination of human amniotic fluid metabolite concentrations, which are then used in a metabonomic study to establish patient health during gestation. {sup 1}H NMR free inductive decays (FIDs) of 258 human amniotic fluid samples from a 500MHz spectrometer are acquired. Quantitative analyses methods in both the frequency- and time-domain are carried out and compared. Frequency-domain analysis is accomplished by integration of the metabolite peaks before and after the inclusion of a known standard addition of alanine. Time-domain analysis is accomplished by the direct exponential curve resolution algorithm (DECRA). Both techniques are assessed by applications to calibration biological solutions and a simulated data set. The DECRA method proves to be a more accurate and precise route for quantitative analysis, and is included in the developed protocol. Well-defined peaks of various components are visible in the frequency-domain {sup 1}H NMR spectra, including lactate, alanine, acetate, citrate, choline, glycine, and glucose. All are quantified with the proposed protocol. Statistical t-test and notched box and whisker plots are used to compare means of metabolite concentrations for diabetic and normal patients. Glucose, glycine, and choline are all found to correlate with gestational diabetes mellitus early in gestation. With further development, time-domain quantitative {sup 1}H NMR has potential to become a robust diagnostic tool for gestational health. (author)

  9. Purification and antimicrobial activity studies of the N-terminal fragment of ubiquitin from human amniotic fluid.

    Science.gov (United States)

    Kim, Jin-Young; Lee, Sun Young; Park, Seong-Cheol; Shin, Song Yub; Choi, Sang Joon; Park, Yoonkyung; Hahm, Kyung-Soo

    2007-09-01

    A 4.3-kDa antimicrobial peptide was isolated from human amniotic fluid by dialysis, ultrafiltration, and C18 reversed-phase high performance liquid chromatography. This peptide, which we named Amniotic Fluid Peptide-1 (AFP-1), possessed antimicrobial activity but lacked hemolytic activity. In addition, AFP-1 potently inhibited the growth of a variety of bacteria (Escherichia coli, Salmonella typhimurium, Listeria monocytogenes and Staphylococcus aureus), filamentous fungi (Botrytis cinerea, Aspergillus fumigatus, Neurospora crassa and Fusarium oxysporum) and yeast cells (Candida albicans and Cryptococcus neoformans). Automated Edman degradation showed that the N-terminal sequence of AFP-1 was NH(2)-Met-Gln-Ile-Phe-Val-Lys-Thr-Leu-Thr-Gly-Lys-Thr-Ile-Thr-Leu-Glu-Val-Glu-. The partial sequence had 100% homology to the N-terminal sequence of ubiquitin. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that the molecular mass of AFP-1 was 4280.2 Da. Our data show an antimicrobial activity of ubiquitin N-terminal derived peptide that makes it suitable for use as an antimicrobial agent.

  10. Eicosanomic profiling reveals dominance of the epoxygenase pathway in human amniotic fluid at term in spontaneous labor.

    Science.gov (United States)

    Maddipati, Krishna Rao; Romero, Roberto; Chaiworapongsa, Tinnakorn; Zhou, Sen-Lin; Xu, Zhonghui; Tarca, Adi L; Kusanovic, Juan Pedro; Munoz, Hernan; Honn, Kenneth V

    2014-11-01

    Lipid mediators play an important role in reproductive biology, especially, in parturition. Enhanced biosynthesis of eicosanoids, such as prostaglandin E2 (PGE2) and PGF2α, precedes the onset of labor as a result of increased expression of inducible cyclooxygenase 2 (COX-2) in placental tissues. Metabolism of arachidonic acid results in bioactive lipid mediators beyond prostaglandins that could significantly influence myometrial activity. Therefore, an unbiased lipidomic approach was used to profile the arachidonic acid metabolome of amniotic fluid. In this study, liquid chromatography-mass spectrometry was used for the first time to quantitate these metabolites in human amniotic fluid by comparing patients at midtrimester, at term but not in labor, and at term and in spontaneous labor. In addition to exposing novel aspects of COX pathway metabolism, this lipidomic study revealed a dramatic increase in epoxygenase- and lipoxygenase-pathway-derived lipid mediators in spontaneous labor with remarkable product selectivity. Despite their recognition as anti-inflammatory lipid mediators and regulators of ion channels, little is known about the epoxygenase pathway in labor. Epoxygenase pathway metabolites are established regulators of vascular homeostasis in cardiovascular and renal physiology. Their presence as the dominant lipid mediators in spontaneous labor at term portends a yet undiscovered physiological function in parturition. © FASEB.

  11. Diagnóstico laboratorial do líquido amniótico Laboratory diagnosis of amniotic fluid

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    Sabrina Gonçalves Campana

    2003-09-01

    Full Text Available O presente trabalho tem como objetivos a definição e a fisiologia do líquido amniótico, ressaltando aspectos citológicos e principais técnicas para diagnóstico laboratorial das patologias mais freqüentes. A metodologia utilizada foi a revisão bibliográfica atualizada relacionando os aspectos citológicos com a idade gestacional e técnicas laboratoriais para diagnóstico das principais patologias em que são observadas alterações do líquido amniótico, concluindo-se que este é um importante componente do ambiente intra-uterino. Sua produção e absorção dependem de uma série de mecanismos interdependentes entre o feto, a placenta, as membranas e o organismo materno. Atualmente este fluido pode fornecer inúmeras informações sobre a saúde fetal, realizando-se diversas técnicas, entre elas a amniocentese e a dosagem de alfafetoproteína, que pode detectar defeitos do tubo neural e trissomia do cromossomo 21. A análise do líquido amniótico reforça a importância da realização adequada de um pré-natal, sendo importante relacionar os resultados laboratoriais com a clínica.This present paper aims the definition of the amniotic fluid and its physiology standing out cytological aspects and main techniques for laboratorial diagnosis of the most frequent pathologies. The methodology was based on updated bibliographical research relating the cytological aspects with the pregnancy age and laboratorial techniques for diagnosis of the main pathologies in which alterations of the amniotic fluid are observed, concluding that this is an important component of the intrauterine environment. Its production and absorption depend on a series of interdependent mechanisms among the fetus, the placenta, the membranes and the maternal organism. Currently this fluid can supply innumerable information on the fetal health by the use of diverse techniques, among which, amniocentesis and dosage of alpha-fetoprotein, which can detect defects of the

  12. A prenatal test for the cerebro-hepato-renal (Zellweger) syndrome by demonstration of the absence of catalase-containing particles (peroxisomes) in cultured amniotic fluid cells

    NARCIS (Netherlands)

    Wanders, R. J.; Schrakamp, G.; van den Bosch, H.; Tager, J. M.; Schutgens, R. B.

    1986-01-01

    In this paper we show that whereas acyl-CoA: dihydroxyacetone phosphate acyltransferase, a membrane-bound peroxisomal enzyme, is deficient in homogenates of cultured amniotic fluid cells of fetuses with Zellweger syndrome, catalase a soluble peroxisomal matrix enzyme is present in normal amounts.

  13. Effects of Having Preterm Infants Smell Amniotic Fluid, Mother's Milk, and Mother's Odor During Heel Stick Procedure on Pain, Physiological Parameters, and Crying Duration.

    Science.gov (United States)

    Küçük Alemdar, Dilek; Kardaş Özdemir, Funda

    2017-06-01

    The study aims to assess effects of having preterm infants smell amniotic fluid, mother's milk, and mother's odor during heel stick procedure on pain, physiological parameters, and crying duration. The study was a randomized controlled experimental research. The population of the study was made up of preterm infants receiving treatment and care at a neonatal intensive care unit, where the study was conducted between January 2015 and March 2016. The study was performed with 85 preterm infants who met the selection criteria. Infants were randomized into four groups: amniotic fluid, mother's milk, mother's odor, and control group. Data obtained were analyzed by percentage distributions, means, standard deviation, chi-square test, Kruskal-Wallis, and Dunnett's test. While no significant difference was found between the groups in terms of total preterm infant pain profile score before, during, and after the heel stick procedure (p > 0.05), a significant difference was found in terms of SO 2 values (p  0.05), the amniotic fluid group had the lowest score, followed by the mother's milk group, the mother's odor group, and the control group. Amniotic fluid, mother's milk, and mother's odor were not effective in preterm infants during painful procedures.

  14. Amniotic fluid l-ergothioneine concentrations in pregnant sheep after natural mating and transfer of vitrified/thawed in-vitro produced embryos.

    Science.gov (United States)

    Sotgia, Salvatore; Zinellu, Angelo; Arru, Dionigia; Nieddu, Stefano; Strina, Alessandro; Ariu, Federica; Pintus, Gianfranco; Carru, Ciriaco; Bogliolo, Luisa; Ledda, Sergio

    2015-10-01

    L-ergothioneine levels were measured in amniotic fluid of pregnant sheep after natural mating and transfer of vitrified/thawed in-vitro produced embryos. Amniotic fluids were collected between 60 and 65 and 80-85 days of gestation and analysed by an ultra-performance liquid chromatographic (UPLC)method with fluorescence detection. L-Ergothioneine concentrations ranged between 0.23 and 9.36 μmol/L and were significantly higher in pregnancy obtained by the transfer of vitrified/thawed in-vitro produced embryos. Conversely, no significant changes in amniotic fluid L-ergothioneine concentrations were observed according to the stages of pregnancy considered in this study. These findings suggest that L-ergothioneine concentrations, are not affected as much by the gestational age, but rather by the method used to induce the pregnancy. On the whole, the measurement of L-ergothioneine in amniotic fluid could serve as a useful biomarker of oxidative stress and/or inflammatory state in pregnancy.

  15. The Amniotic Membrane: Development and Potential Applications - A Review.

    Science.gov (United States)

    Favaron, P O; Carvalho, R C; Borghesi, J; Anunciação, A R A; Miglino, M A

    2015-12-01

    Foetal membranes are essential tissues for embryonic development, playing important roles related to protection, breathing, nutrition and excretion. The amnion is the innermost extraembryonic membrane, which surrounds the foetus, forming an amniotic sac that contains the amniotic fluid (AF). In recent years, the amniotic membrane has emerged as a potential tool for clinical applications and has been primarily used in medicine in order to stimulate the healing of skin and corneal diseases. It has also been used in vaginal reconstructive surgery, repair of abdominal hernia, prevention of surgical adhesions and pericardium closure. More recently, it has been used in regenerative medicine because the amniotic-derived stem cells as well as AF-derived cells exhibit cellular plasticity, angiogenic, cytoprotective, immunosuppressive properties, antitumoural potential and the ability to generate induced pluripotent stem cells. These features make them a promising source of stem cells for cell therapy and tissue engineering. In this review, we discussed the development of the amnion, AF and amniotic cavity in different species, as well as the applicability of stem cells from the amnion and AF in cellular therapy. © 2015 Blackwell Verlag GmbH.

  16. [Maternal deaths due to amniotic fluid embolism. Results from the French confidential enquiry into maternal deaths, 2010-2012].

    Science.gov (United States)

    Morau, E; Proust, A; Ducloy, J-C

    2017-12-01

    Amniotic fluid embolism (AFE) is an unpredictable, dreadful complication of pregnancy or childbirth. EA typically includes in the same lapse of time respiratory, haemodynamic, neurological and hemorrhagic symptoms (from early and severe coagulopathy). Immediate supportive treatment by a multidisciplinary team is the cornerstone of the management. Between 2010 an 2012 in France, 24 deaths were related to AFE giving a maternal mortality ratio of 1/100,000 live births (CI 95% 0.6-1.4). AFE ranks as the second leading cause of direct maternal death. Eight cases over 23 were classified as having some degree of substandard care. Substandard care included delays in performing aggressive surgical treatment or delays in the diagnosis and the treatment of the coagulopathy. Learning points focus on the importance to pay attention on premonitory symptoms, to early assess the clotting status and to train in multidisciplinary team. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  17. MicroRNA Expression Profile in the Prenatal Amniotic Fluid Samples of Pregnant Women with Down Syndrome.

    Science.gov (United States)

    Karaca, Emin; Aykut, Ayça; Ertürk, Biray; Durmaz, Burak; Güler, Ahmet; Büke, Barış; Yeniel, Ahmet Özgür; Ergenoğlu, Ahmet Mete; Özkınay, Ferda; Özeren, Mehmet; Kazandı, Mert; Akercan, Fuat; Sağol, Sermet; Gündüz, Cumhur; Çoğulu, Özgür

    2018-03-15

    Down syndrome, which is the most common human chromosomal anomaly that can affect people of any race and age, can be diagnosed prenatally in most cases. Prenatal diagnosis via culture method is time-consuming; thus, genetic analysis has thus been introduced and is continually being developed for rapid prenatal diagnosis. For this reason, the effective use of microRNA profiling for the rapid analysis of prenatal amniotic fluid samples for the diagnosis of Down syndrome was investigated. To evaluate the expression levels of 14 microRNAs encoded by chromosome 21 in amniotic fluid samples and their utility for prenatal diagnosis of Down syndrome. Case-control study. We performed invasive prenatal testing for 56 pregnant women; 23 carried fetuses with Down syndrome, and 33 carried fetuses with a normal karyotype. Advanced maternal age and increased risk for Down syndrome in the screening tests were indications for invasive prenatal testing. The age of gestation in the study and control groups ranged between 17 and 18 weeks. The expression levels of microRNA were measured by real-time polymerase chain reaction. The expression levels of microRNA-125b-2, microRNA-155 , and microRNA-3156 were significantly higher in the study group than in the control group. The presence of significantly dysregulated microRNAs may be associated with either the phenotype or the result of abnormal development. Further large-scale comparative studies conducted in a variety of conditions may bring novel insights in the field of abnormal prenatal conditions.

  18. Fetal Alcohol Syndrome (FAS) in C57BL/6 Mice Detected through Proteomics Screening of the Amniotic Fluid

    Science.gov (United States)

    Datta, Susmita; Turner, Delano; Singh, Reetu; Ruest, L. Bruno; Pierce, William M.; Knudsen, Thomas B.

    2009-01-01

    BACKGROUND Fetal Alcohol Syndrome (FAS), a severe consequence of the Fetal Alcohol Spectrum Disorders, is associated with craniofacial defects, mental retardation, and stunted growth. Previous studies in C57BL/6J and C57BL/6N mice provide evidence that alcohol-induced pathogenesis follows early changes in gene expression within specific molecular pathways in the embryonic headfold. Whereas the former (B6J) pregnancies carry a high-risk for dysmorphogenesis following maternal exposure to 2.9 g/kg alcohol (two injections spaced 4.0 h apart on gestation day 8), the latter (B6N) pregnancies carry a low-risk for malformations. The present study used this murine model to screen amniotic fluid for biomarkers that could potentially discriminate between FAS-positive and FAS-negative pregnancies. METHODS B6J and B6N litters were treated with alcohol (exposed) or saline (control) on day 8 of gestation. Amniotic fluid aspirated on day 17 (n = 6 replicate litters per group) was subjected to trypsin digestion for analysis by matrix-assisted laser desorption–time of flight mass spectrometry with the aid of denoising algorithms, statistical testing, and classification methods. RESULTS We identified several peaks in the proteomics screen that were reduced consistently and specifically in exposed B6J litters. Preliminary characterization by liquid chromatography tandem mass spectrometry and multidimensional protein identification mapped the reduced peaks to alpha fetoprotein (AFP). The predictive strength of AFP deficiency as a biomarker for FAS-positive litters was confirmed by area under the receiver operating characteristic curve. CONCLUSIONS These findings in genetically susceptible mice support clinical observations in maternal serum that implicate a decrease in AFP levels following prenatal alcohol damage. PMID:18240165

  19. Diagnostic Role of MicroRNA Expression Profile in the Prenatal Amniotic Fluid Samples of Pregnant Women with Down Syndrome

    Directory of Open Access Journals (Sweden)

    Emin Karaca

    2018-03-01

    Full Text Available Background: Down syndrome, which is the most common human chromosomal anomaly that can affect people of any race and age, can be diagnosed prenatally in most cases. Prenatal diagnosis via culture method is time-consuming; thus, genetic analysis has thus been introduced and is continually being developed for rapid prenatal diagnosis. For this reason, the effective use of microRNA profiling for the rapid analysis of prenatal amniotic fluid samples for the diagnosis of Down syndrome was investigated. Aims: To evaluate the expression levels of 14 microRNAs encoded by chromosome 21 in amniotic fluid samples and their utility for prenatal diagnosis of Down syndrome. Study Design: Case-control study. Methods: We performed invasive prenatal testing for 56 pregnant women; 23 carried fetuses with Down syndrome, and 33 carried fetuses with a normal karyotype. Advanced maternal age and increased risk for Down syndrome in the screening tests were indications for invasive prenatal testing. The age of gestation in the study and control groups ranged between 17 and 18 weeks. The expression levels of microRNA were measured by real-time polymerase chain reaction. Results: The expression levels of microRNA-125b-2, microRNA-155, and microRNA-3156 were significantly higher in the study group than in the control group. Conclusion: The presence of significantly dysregulated microRNAs may be associated with either the phenotype or the result of abnormal development. Further large-scale comparative studies conducted in a variety of conditions may bring novel insights in the field of abnormal prenatal conditions.

  20. Genital tract, cord blood, and amniotic fluid exposures of seven antiretroviral drugs during and after pregnancy in human immunodeficiency virus type 1-infected women.

    Science.gov (United States)

    Yeh, Rosa F; Rezk, Naser L; Kashuba, Angela D M; Dumond, Julie B; Tappouni, Hiba L; Tien, Hsiao-Chuan; Chen, Ya-Chi; Vourvahis, Manoli; Horton, Amanda L; Fiscus, Susan A; Patterson, Kristine B

    2009-06-01

    The objective of the study was to measure antiretroviral exposures in four physiological compartments during pregnancy, delivery, and postpartum. This prospective, open-label, longitudinal study collected paired blood plasma (BP) and genital tract (GT) aspirates antepartum, at delivery, and up to 12 weeks postpartum. Antiretroviral cord BP and amniotic fluid concentrations were also measured. Drug concentrations were analyzed by validated high-performance liquid chromatography/UV and liquid chromatography/tandem mass spectrometry methods, with secondary compartment concentrations presented as the percentage of BP. Fourteen women taking lamivudine plus zidovudine and either lopinavir-ritonavir (n = 7), nelfinavir (n = 6), or nevirapine (n = 1) were enrolled; four also received tenofovir. GT penetration relative to BP was highest for the nucleoside reverse transcriptase inhibitors compared to the protease inhibitors and nevirapine. Only antepartum nelfinavir GT penetration was significantly higher than in the second trimester (geometric mean ratio [GMR], 179.3) or third trimester (GMR, 41.9). Compared to nonpregnant historical controls, antepartum GT penetration was significantly lower (P or = 100%), with cord BP levels of the remaining drugs ranging from 49 to 86% of that of the respective BP level. Amniotic exposures for lamivudine, zidovudine, tenofovir, and nelfinavir were > or = 100%, nevirapine exposure was 53%, and lopinavir and ritonavir exposures were amniotic fluid exposures vary within and between antiretroviral drug classes and biologic sites. Measurement of antiretroviral exposure in maternal genital secretions, cord BP, and amniotic fluid may be needed to identify signals of subtherapeutic or supratherapeutic drug exposure.

  1. Amniotic mesenchymal stem cells enhance wound healing in diabetic NOD/SCID mice through high angiogenic and engraftment capabilities.

    Science.gov (United States)

    Kim, Sung-Whan; Zhang, Hong-Zhe; Guo, Longzhe; Kim, Jong-Min; Kim, Moo Hyun

    2012-01-01

    Although human amniotic mesenchymal stem cells (AMMs) have been recognised as a promising stem cell resource, their therapeutic potential for wound healing has not been widely investigated. In this study, we evaluated the therapeutic potential of AMMs using a diabetic mouse wound model. Quantitative real-time PCR and ELISA results revealed that the angiogenic factors, IGF-1, EGF and IL-8 were markedly upregulated in AMMs when compared with adipose-derived mesenchymal stem cells (ADMs) and dermal fibroblasts. In vitro scratch wound assays also showed that AMM-derived conditioned media (CM) significantly accelerated wound closure. Diabetic mice were generated using streptozotocin and wounds were created by skin excision, followed by AMM transplantation. AMM transplantation significantly promoted wound healing and increased re-epithelialization and cellularity. Notably, transplanted AMMs exhibited high engraftment rates and expressed keratinocyte-specific proteins and cytokeratin in the wound area, indicating a direct contribution to cutaneous closure. Taken together, these data suggest that AMMs possess considerable therapeutic potential for chronic wounds through the secretion of angiogenic factors and enhanced engraftment/differentiation capabilities.

  2. Transplantation of human umbilical cord blood or amniotic epithelial stem cells alleviates mechanical allodynia after spinal cord injury in rats.

    Science.gov (United States)

    Roh, Dae-Hyun; Seo, Min-Soo; Choi, Hoon-Seong; Park, Sang-Bum; Han, Ho-Jae; Beitz, Alvin J; Kang, Kyung-Sun; Lee, Jang-Hern

    2013-01-01

    Stem cell therapy is a potential treatment for spinal cord injury (SCI), and a variety of different stem cell types have been grafted into humans suffering from spinal cord trauma or into animal models of spinal injury. Although several studies have reported functional motor improvement after transplantation of stem cells into injured spinal cord, the benefit of these cells for treating SCI-induced neuropathic pain is not clear. In this study, we investigated the therapeutic effect of transplanting human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) or amniotic epithelial stem cells (hAESCs) on SCI-induced mechanical allodynia (MA) and thermal hyperalgesia (TH) in T13 spinal cord hemisected rats. Two weeks after SCI, hUCB-MSCs or hAESCs were transplanted around the spinal cord lesion site, and behavioral tests were performed to evaluate changes in SCI-induced MA and TH. Immunohistochemical and Western blot analyses were also performed to evaluate possible therapeutic effects on SCI-induced inflammation and the nociceptive-related phosphorylation of the NMDA NR1 receptor subunit. While transplantation of hUCB-MSCs showed a tendency to reduce MA, transplantation of hAESCs significantly reduced MA. Neither hUCB-MSC nor hAESC transplantation had any effect on SCI-induced TH. Transplantation of hAESCs also significantly reduced the SCI-induced increase in NMDA receptor NR1 subunit phosphorylation (pNR1) expression in the spinal cord. Both hUCB-MSCs and hAESCs reduced the SCI-induced increase in spinal cord expression of the microglial marker, F4/80, but not the increased expression of GFAP or iNOS. Taken together, these findings demonstrate that the transplantation of hAESCs into the injured spinal cord can suppress mechanical allodynia, and this effect seems to be closely associated with the modulation of spinal cord microglia activity and NR1 phosphorylation.

  3. The Effects of Intravenous Hydration on Amniotic Fluid Index in Pregnant Women with Preterm premature Rupture of Membranes: A Randomized Clinical Trial

    Science.gov (United States)

    Shahnazi, Mahnaz; Tagavi, Simin; Hajizadeh, Khadije; Farshbaf Khalili, Azize

    2013-01-01

    Introduction: Preterm premature rupture of membranes (PPROM) can result in fetal complications such as oligohydramnios. This study aimed to determine the effects of intravenous (IV) fluid bolus on amniotic fluid index (AFI) in pregnant women with PPROM. Methods: 24 women with PPROM during singleton live pregnancy of 28 to 34 weeks whose baseline AFI was ≤ 5cm were randomized into two groups. The study group received one liter intravenous fluid bolus of isotonic Ringer serum during 30-minute period. Reevaluations of amniotic fluid index in both groups were made 90 minutes and 48 hours after baseline measurement. Independent t-test and paired t-test were used to compare the two groups and mean amniotic fluid index before and after treatment, respectively. Results: The results of this study demonstrate that AFI decreased statistically significant in both the control and study groups. AFI decreased in both groups at 48 hours later. This decrease was not statistically significant in any group. The mean change in AFI (90 minutes and baseline) and (48 hours and baseline) between the two groups were not significant. The time between mean baseline measurements and delivery were 196.41 and 140.58 hours in the study and control groups, respectively. This difference was not statistically significant. Conclusion: This study did not find significant impact of hydration On AFI as a prophylactic method on oligohydramnios in pregnant women with PPROM. PMID:25276709

  4. Clinical significance of histologic chorioamnionitis with a negative amniotic fluid culture in patients with preterm labor and premature membrane rupture.

    Science.gov (United States)

    Park, Jeong Woo; Park, Kyo Hoon; Jung, Eun Young

    2017-01-01

    To evaluate the effect of histological chorioamnionitis (HCA) with a negative amniotic fluid (AF) culture on adverse pregnancy and neonatal outcomes and inflammatory status in the AF compartment in women with preterm labor or preterm premature rupture of membranes (PPROM). This is a retrospective cohort study of 153 women diagnosed as having a preterm labor or PPROM (20-34 weeks) who delivered singleton gestations within 48 hours of amniocentesis. AF obtained through amniocentesis was cultured, and interleukin (IL)-6, IL-8, and metalloproteinase-9 (MMP-9) levels were determined. The placentas were examined histologically. The prevalence of HCA with negative AF culture was 23.5% (36/153). The women with HCA but with a negative AF culture (group 2) and those with a positive AF culture (group 3) had a significantly lower mean gestational age at amniocentesis and delivery than those with a negative AF culture and without HCA (group 1). Women in group 3 had the highest levels of AF IL-6, IL-8, and MMP-9, followed by those in group 2, and those in group 1. Composite neonatal morbidity was significantly higher in groups 2 and 3 than in group 1, but this was no longer significant after adjusting for confounders caused mainly by the impact of gestational age. In the women who delivered preterm neonates, HCA with a negative AF culture was associated with increased risks of preterm birth, intense intra-amniotic inflammatory response, and prematurity-associated composite neonatal morbidity, and its risks are similar to the risk posed by positive AF culture.

  5. Clinical significance of histologic chorioamnionitis with a negative amniotic fluid culture in patients with preterm labor and premature membrane rupture.

    Directory of Open Access Journals (Sweden)

    Jeong Woo Park

    Full Text Available To evaluate the effect of histological chorioamnionitis (HCA with a negative amniotic fluid (AF culture on adverse pregnancy and neonatal outcomes and inflammatory status in the AF compartment in women with preterm labor or preterm premature rupture of membranes (PPROM.This is a retrospective cohort study of 153 women diagnosed as having a preterm labor or PPROM (20-34 weeks who delivered singleton gestations within 48 hours of amniocentesis. AF obtained through amniocentesis was cultured, and interleukin (IL-6, IL-8, and metalloproteinase-9 (MMP-9 levels were determined. The placentas were examined histologically.The prevalence of HCA with negative AF culture was 23.5% (36/153. The women with HCA but with a negative AF culture (group 2 and those with a positive AF culture (group 3 had a significantly lower mean gestational age at amniocentesis and delivery than those with a negative AF culture and without HCA (group 1. Women in group 3 had the highest levels of AF IL-6, IL-8, and MMP-9, followed by those in group 2, and those in group 1. Composite neonatal morbidity was significantly higher in groups 2 and 3 than in group 1, but this was no longer significant after adjusting for confounders caused mainly by the impact of gestational age.In the women who delivered preterm neonates, HCA with a negative AF culture was associated with increased risks of preterm birth, intense intra-amniotic inflammatory response, and prematurity-associated composite neonatal morbidity, and its risks are similar to the risk posed by positive AF culture.

  6. Anti-aging Effect of Transplanted Amniotic Membrane Mesenchymal Stem Cells in a Premature Aging Model of Bmi-1 Deficiency

    Science.gov (United States)

    Xie, Chunfeng; Jin, Jianliang; Lv, Xianhui; Tao, Jianguo; Wang, Rong; Miao, Dengshun

    2015-01-01

    To determine whether transplanted amniotic membrane mesenchymal stem cells (AMSCs) ameliorated the premature senescent phenotype of Bmi-1-deficient mice, postnatal 2-day-old Bmi-1−/− mice were injected intraperitoneally with the second-passage AMSCs from amniotic membranes of β-galactosidase (β-gal) transgenic mice or wild-type (WT) mice labeled with DiI. Three reinjections were given, once every seven days. Phenotypes of 5-week-old β-gal+ AMSC-transplanted or 6-week-old DiI+ AMSC-transplanted Bmi-1−/− mice were compared with vehicle-transplanted Bmi-1−/− and WT mice. Vehicle-transplanted Bmi-1−/− mice displayed growth retardation and premature aging with decreased cell proliferation and increased cell apoptosis; a decreased ratio and dysmaturity of lymphocytic series; premature osteoporosis with reduced osteogenesis and increased adipogenesis; redox imbalance and DNA damage in multiple organs. Transplanted AMSCs carried Bmi-1 migrated into multiple organs, proliferated and differentiated into multiple tissue cells, promoted growth and delayed senescence in Bmi-1−/− transplant recipients. The dysmaturity of lymphocytic series were ameliorated, premature osteoporosis were rescued by promoting osteogenesis and inhibiting adipogenesis, the oxidative stress and DNA damage in multiple organs were inhibited by the AMSC transplantation in Bmi-1−/− mice. These findings indicate that AMSC transplantation ameliorated the premature senescent phenotype of Bmi-1-deficient mice and could be a novel therapy to delay aging and prevent aging-associated degenerative diseases. PMID:26370922

  7. Phospholipid profile of amniotic fluid in ovine model of congenital diaphragmatic hernia (CDH): the effect of fetal tracheal occlusion.

    Science.gov (United States)

    Mimmi, Maria Chiara; Ballico, Maurizio; Amoroso, Francesco; Calcaterra, Valeria; Marotta, Mario; Peiro, Jose Luis; Pelizzo, Gloria

    2015-03-06

    Fetal endoscopic tracheal occlusion has been proposed as a prenatal intervention to ameliorate congenital diaphragmatic hernia (CDH) prognosis. Tracheal occlusion (TO) prevents pulmonary fluid egress, leading to tissue expansion, reversal of lung hypoplasia, and potential maturation. Fetal lung maturity strongly correlates with amniotic fluid (AF) phospholipidic composition. In this preliminary study, we characterized the AF phospholipidic profile in CDH-induced, TO-treated, and healthy fetal lambs to define the prenatal treatment benefits of TO on lung maturity. CDH induction was performed at 70 days of gestation, TO was carried out at 102 days of gestation, and caesarean section was carried out at 136 days of gestation. AF samples, taken at 102-136 days of gestation, were evaluated using mass spectrometry. The analysis focused on phosphatidylcholines (PCs) and sphingomyelins (SMs). The most abundant phosphatidylcholine species retrieved in healthy AF was POPC [PC(18:1/16:0)], while the level of DPPC [PC(16:0/16:0)] was extremely low at both gestational ages. CDH induction caused a decrease in POPC and many other PCs. A substantial return of some PCs, in particular POPC, PC(34:2) and PC(18:0/16:0), to a more physiological level was prompted by TO. SMs were unaltered. The AF phospholipidic profile could provide prenatal prognostic markers of CDH and possible indices of lung maturation after fetal treatment.

  8. Evaluation of five automated and one manual method for Toxoplasma and human DNA extraction from artificially spiked amniotic fluid.

    Science.gov (United States)

    Yera, H; Ménégaut, L; Brenier-Pinchart, M-P; Touafek, F; Bastien, P; Dalle, F

    2018-01-31

    Molecular detection of Toxoplasma gondii plays a crucial role in the prenatal and neonatal diagnosis of congenital toxoplasmosis (CT). Sensitivity of this diagnosis is partly related to the efficiency of parasite DNA extraction and amplification. DNA extraction methods with automated platforms have been developed. Therefore, it is essential to evaluate them in combination with adequate PCR amplification assays. In this multisite study, we investigated the suitability of two recent automated procedures for the isolation of Toxoplasma DNA from amniotic fluid (AF) (Magtration system 12GC, PSS and Freedom EVO VacS, Tecan), compared with three other automated procedures (MagNAPure Compact, Roche, BioRobot EZ1, Qiagen and modified NucliSens easyMAG, bioMérieux) and with the manual DNA extraction QIAamp DNA Mini kit (Qiagen). Two Toxoplasma PCR assays targeting the '529-bp' repeat DNA element were used, based upon dual hybridization (FRET) or hydrolysis (TaqMan) probes. A total of 1296 PCRs were performed including 972 Toxoplasma PCRs. We showed variable efficacy (4.2%-100% positive results) among the DNA extraction procedures in isolating up to five T. gondii cells/mL in AF samples. Moreover, for a given DNA extraction method, variable results were obtained among the two Toxoplasma PCR assays for detecting up to five T. gondii cells/mL: when using TaqMan PCR, all the automated systems yielded more than 60% positive results. Nevertheless, when testing the DNA extracts in triplicate, four out of six extraction methods allowed a satisfactory detection of low amounts of T. gondii DNA (≥33% of positive results) independently of the PCR assay used. Despite the influence of the subsequent PCR method used, this study should help microbiologists in the choice of DNA extraction methods for the detection of T. gondii in amniotic fluid. The extraction method should be checked as adequate for the PCR assay used. Copyright © 2018 European Society of Clinical Microbiology and

  9. Carcinogenicity, efficiency and biosafety analysis in xeno-free human amniotic stem cells for regenerative medical therapies.

    Science.gov (United States)

    Phermthai, Tatsanee; Thongbopit, Sasiprapa; Pokathikorn, Puttachart; Wichitwiengrat, Suparat; Julavijitphong, Suphakde; Tirawanchai, Nednapis

    2017-08-01

    Human amniotic mesenchymal stromal cells (hAMSCs) are a potent and attractive stem cell source for use in regenerative medicine. However, the safe uses of therapeutic-grade MSCs are equally as important as the efficiency of MSCs. To provide efficient, clinic-compliant (safe for therapeutic use) MSCs, hAMSC lines that completely eliminate the use of animal products and have been characterized for carcinogenicity and biosafety are required. Here, we have efficiently generated 10 hAMSC lines under human umbilical cord blood serum (hUCS)-supplemented medium (xeno-free culture) and fetal bovine serum (FBS)-supplemented medium (standard culture) and investigated carcinogenicity and immunosuppressive properties in the resultant hAMSC lines. All hAMSC lines were examined for efficiency (growth kinetics, cryopreservation, telomere length, phenotypic characterization, differentiation potential), carcinogenicity (proto-oncogene and tumor suppressor gene and epigenomic stability) and safety (immunosuppressive properties). Stem cell characteristics between the xeno-free hAMSC lines and the cell lines generated using the standard culture system showed no differences. Xeno-free hAMSC lines displayed normal growth proliferation potential, morphological, karyotypic, phenotypic differentiation properties and telomere lengths. Additionally, they retained normal immunosuppressive effects. As a marker of carcinogenicity and biosafety, proto-oncogenes expression levels showed no differences in xeno-free hAMSCs, and we detected no SNP mutations on hotspot codons of the P53 tumor suppressor gene and stable epigenomic imprinting in xeno-free hAMSC lines. Xeno-free hAMSC lines retain essential stem cell characteristics, with a high degree of certainty for meeting biosafety and carcinogenicity standards for a xeno-free system supplemented with allogenic hUCS. The cell lines are suitable and valuable for therapeutic purposes. Copyright © 2017 International Society for Cellular Therapy

  10. Perfluorooctane Sulfonate Concentrations in Amniotic Fluid, Biomarkers of Fetal Leydig Cell Function, and Cryptorchidism and Hypospadias in Danish Boys (1980-1996)

    DEFF Research Database (Denmark)

    Toft, Gunnar; Jönsson, Bo A; Bonde, Jens P

    2016-01-01

    BACKGROUND: Exposure to perfluorooctane sulfonate (PFOS) may potentially disturb fetal Leydig cell hormone production and male genital development. OBJECTIVES: We aimed to study the associations between levels of amniotic fluid PFOS, fetal steroid hormone, and insulin-like factor 3 (INSL3) and th...... Leydig cell function, and cryptorchidism and hypospadias in Danish boys (1980-1996). Environ Health Perspect 124:151-156; http://dx.doi.org/10.1289/ehp.1409288....

  11. Analysis of nifedipine in human plasma and amniotic fluid by liquid chromatography-tandem mass spectrometry and its application to clinical pharmacokinetics in hypertensive pregnant women.

    Science.gov (United States)

    Filgueira, Gabriela Campos de Oliveira; Filgueira, Osmany Alberto Silva; Carvalho, Daniela Miarelli; Marques, Maria Paula; Moisés, Elaine Christine Dantas; Duarte, Geraldo; Lanchote, Vera Lucia; Cavalli, Ricardo Carvalho

    2015-07-01

    Nifedipine is a dihydropyridine calcium channel blocker used for the treatment of hypertension in pregnant women. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for analysis of nifedipine in human plasma and amniotic fluid. Separation of nifedipine and nitrendipine (IS) was performed using a LiChroCART(®) RP-Select B column and a mixture of water:acetonitrile:glacial acetic acid (30:70:0.5 v/v) as the mobile phase. Aliquots of 500μL of biological samples were extracted at pH 13 using dichloromethane:n-pentane (3:7 v/v). The validated method was applied to a study of the pharmacokinetics of nifedipine in human plasma and amniotic fluid samples collected up to 12h after administration of the last slow-release nifedipine (20mg/12h) dose to 12 hypertensive pregnant women. The estimated pharmacokinetic parameters of nifedipine showed a mean AUC(0-12) of 250.2ngh/mL, ClT/F of 89.2L/h, Vd/F of 600.0L and t1/2 5.1h. The mean amniotic fluid/plasma concentration ratio was 0.05. The methods proved to be highly sensitive by showing a lower quantification limit of 0.1ng/mL for both matrices. And this study reports for the first time the complete development and validation of the method to quantify nifedipine in amniotic fluid using LC-MS-MS. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Maternal-fetal transfer and amniotic fluid accumulation of nucleoside analogue reverse transcriptase inhibitors in human immunodeficiency virus-infected pregnant women.

    Science.gov (United States)

    Chappuy, Hélène; Tréluyer, Jean-Marc; Jullien, Vincent; Dimet, Jérôme; Rey, Elisabeth; Fouché, Maria; Firtion, Ghislaine; Pons, Gérard; Mandelbrot, Laurent

    2004-11-01

    This study was performed to investigate placental transfer of nucleoside analogue reverse transcriptase inhibitors (NRTIs) and their concentrations in amniotic fluid when given to human immunodeficiency virus (HIV)-infected pregnant women. A total of 100 HIV type 1-infected mothers receiving antiretroviral therapy, including one or more NRTIs, for clinical indications at the time of delivery were enrolled. Maternal blood samples and amniotic fluid were obtained during delivery or cesarean section, and paired cord blood samples were obtained by venipuncture immediately after delivery. Drug concentrations were measured by using high-performance liquid chromatography. A significant relationship between concentrations in maternal and cord plasma samples was found for zidovudine, lamivudine, stavudine, and didanosine. The ratio between the concentrations in cord and maternal plasma samples (R) was high for zidovudine (R = 1.22), its glucuronide metabolite (3'-azido-3'-deoxythymidine-beta-d-glucuronide) (R = 1.01), stavudine (R = 1.32), lamivudine (R = 0.93), and abacavir (R = 1.03) and was low for didanosine (R = 0.38). The ratio between the concentrations in amniotic fluid and cord plasma samples was high for zidovudine (R = 2.24), its glucuronide metabolite (R = 2.83), stavudine (R = 4.87), and lamivudine (R = 3.99) and was lower for didanosine (R = 1.14). These findings indicate that most NRTIs cross the placenta by simple diffusion and are concentrated in the amniotic fluid, probably through fetal urinary excretion. The efficacy or toxicity of NRTIs may vary according to placental transfer.

  13. Utilization of a single antiserum for the direct radioimmunoassay of prostaglandins E and F in semen and prostaglandin F in amniotic fluid

    International Nuclear Information System (INIS)

    Clarke, A.H.; Ing, R.M.Y.; Jones, W.R.; Llewellyn-Jones, D.; Shutt, D.A.

    1974-01-01

    Antibodies to both prostaglandin F (PGF) and prostaglandin E (PGE) were raised in rabbits after they were immunized with prostaglandin F/sub 2a/ conjugated to bovine serum albumin (PGF/sub 2a/--BSA). The antisera were group specific although the antibodies to the F group of prostaglandins showed greater specificity than those to the E group. The antisera were sufficiently specific however to allow the direct radioimmunoassay of PGF and PGE in human semen and PGF in amniotic fluid during induced abortion. Specificity of the direct radioimmunoassay was checked by chromatographic separation of the prostaglandins prior to analysis. Estimation of the prostaglandins in the semen of 30 men attending the infertility clinic showed that 19 of the men had normal semen levels of PGE and PGF of 68 +- 7 (SE) and 6.0 +- 0.6 μg/ml respectively, as compared with data on normal fertile males, whilst the other 11 men had lower levels of 16 +- 2 (SE) and 0.8 +- 0.1 μg/ml respectively. Application of the method to amniotic fluid showed that the PGF concentration in amniotic fluid during the induction of abortion with extra-ovular saline increased from less than 0.6 ng/ml to 6.4 ng/ml when the induction-abortion intervals ranged from 6 to 48 hours. (U.S.)

  14. Non-invasively collected amniotic fluid as a source of possible biomarkers for premature rupture of membranes investigated by proteomic approach.

    Science.gov (United States)

    Consonni, Sara; Mainini, Veronica; Pizzardi, Agnese; Gianazza, Erica; Chinello, Clizia; Locatelli, Anna; Magni, Fulvio

    2014-02-01

    Preterm delivery is one of the main causes of perinatal morbidity and mortality and it accounts for 75 % of perinatal mortality and more than half of the long-term morbidity. We applied a proteomic approach based on mass spectrometry (MS) for biomarkers discovery of preterm premature rupture of membranes (pPROM) by investigating amniotic fluid (AF) invasively and non-invasively collected. Amniotic fluid was obtained from vagina of women with pPROM (group 1), PROM at term (group 2) and by genetic amniocentesis (group 3). Pre-fractionated AF proteome was analyzed through matrix assisted laser desorption ionization-time of flight (MALDI-TOF) MS. The characterization of proteins/peptides of interest was obtained by high performance liquid chromatography-electrospray tandem MS. Three peptides overexpressed in pPROM and able to discriminate the groups 1 and 2 were detected. One peptide was identified as the fragment Gly452LAVPDGPLGLPPKPro466 of the protein KIAA1522, expressed by fetal brain and liver. This peptide was overexpressed in a patient of the group 3, completely asymptomatic at the time of the amniocentesis, who later developed pPROM. Amniotic fluid invasively and non-invasively collected can be analyzed by MALDI-TOF MS to obtain proteomic profiles. Proteomic analysis identified a peptide with promising diagnostic capability for pPROM.

  15. (1)H HR-MAS spectroscopy for quantitative measurement of choline concentration in amniotic fluid as a marker of fetal lung maturity: inter- and intraobserver reproducibility study.

    Science.gov (United States)

    Joe, Bonnie N; Vahidi, Kiarash; Zektzer, Andrew; Chen, Mei-Hsiu; Clifton, Matthew S; Butler, Thomas; Keshari, Kayvan; Kurhanewicz, John; Coakley, Fergus; Swanson, Mark G

    2008-12-01

    To determine the intra- and interobserver reproducibility of human amniotic fluid metabolite concentration measurements (including potential markers of fetal lung maturity) detectable by MR spectroscopy. (1)H high-resolution magic angle spinning (HR-MAS) spectroscopy was performed at 11.7 T on 23 third-trimester amniotic fluid samples. Samples were analyzed quantitatively using 3-(trimethylsilyl)propionic-2,2,3,3-d(4) acid (TSP) as a reference. Four observers independently quantified eight metabolite regions (TSP, lactate doublet and quartet, alanine, citrate, creatinine, choline, and glucose) twice from anonymized, randomized spectra using a semiautomated software program. Excellent inter- and intraobserver reproducibility was found for all metabolites. Intraclass correlation as a measure of interobserver agreement for the four readers ranged from 0.654 to 0.995. A high correlation of 0.973 was seen for choline in particular, a major component of surfactant. Pearson correlation as a measure of intraobserver reproducibility ranged from 0.478 to 0.999. Quantification of choline and other metabolite concentrations in amniotic fluid by high-resolution MR spectroscopy can be performed with high inter- and intraobserver reproducibility. Demonstration of reproducible metabolite concentration measurements is a critical first step in the search for biomarkers of fetal lung maturity. (c) 2008 Wiley-Liss, Inc.

  16. A step-by-step diagnosis of exclusion in a twin pregnancy with acute respiratory failure due to non-fatal amniotic fluid embolism: a case report

    Directory of Open Access Journals (Sweden)

    Papaioannou Vasilios E

    2008-05-01

    Full Text Available Abstract Introduction Respiratory failure may develop during the later stages of pregnancy and is usually associated with tocolysis or other co-existing conditions such as pneumonia, sepsis, pre-eclampsia or amniotic fluid embolism syndrome. Case presentation We present the case of a 34-year-old healthy woman with a twin pregnancy at 31 weeks and 6 days who experienced acute respiratory failure, a few hours after administration of tocolysis (ritodrine, due to preterm premature rupture of the membranes. Her chest discomfort was significantly ameliorated after the ritodrine infusion was stopped and a Cesarean section was performed 48 hours later under spinal anesthesia; however, 2 hours after surgery she developed severe hypoxemia, hypotension, fever and mild coagulopathy. The patient was intubated and transferred to the intensive care unit where she made a quick and uneventful recovery within 3 days. As there was no evidence for drug- or infection-related thromboembolic or myocardial causes of respiratory failure, we conclude that our patient experienced a rare type of non-fatal amniotic fluid embolism. Conclusion In spite of the lack of solid scientific support for our diagnosis, we conclude that our patient suffered an uncommon type of amniotic fluid embolism syndrome and we believe that this report highlights the need for extreme vigilance and a high index of suspicion for such a diagnosis in any pregnant individual.

  17. Array-CGH analysis of cell-free fetal DNA in 10 mL of amniotic fluid supernatant.

    Science.gov (United States)

    Lapaire, Olav; Lu, Xin-Yan; Johnson, Kirby L; Jarrah, Zina; Stroh, Helene; Cowan, Janet M; Tantravahi, Umadevi; Bianchi, Diana W

    2007-07-01

    Previously, we showed that analysis of amniotic fluid (AF) supernatant cell-free fetal (cff) DNA using DNA microarrays (array-CGH) allows for detection of whole chromosome differences between test and reference DNA. Subsequent technical advances have increased both the yield and quality of extracted cffDNA. Here we determined whether array-CGH using smaller volumes of both fresh and frozen AF cffDNA could identify fetal aneuploidy. CffDNA was extracted from 10 mL of residual AF supernatant. The test AF samples (n = 10) included one with a normal karyotype, and nine with the following fetal aneuploidies: trisomies 13 (n = 1), 18 (n = 3), 21 (n = 2), trisomy 9 mosaicism (47,XX,+ 9[18]/46,XX[2]), triploidy (69,XXY) and Turner syndrome (45,X). Array-CGH using AF cffDNA from aneuploid fetuses, compared to euploid reference AF cffDNA, detected whole chromosome aneuploidy in 8 of 9 cases tested, including the case of trisomy 9 mosaicism. The case of triploidy was not detected. CffDNA extracted from 10 mL AF supernatant can be analyzed using array-CGH to correctly identify human chromosome abnormalities. This technology allows for rapid screening of AF samples for whole chromosomal changes by using routinely discarded supernatant, and may augment standard prenatal karyotyping techniques by providing additional molecular information.

  18. Mercury concentrations in human placenta, umbilical cord, cord blood and amniotic fluid and their relations with body parameters of newborns

    International Nuclear Information System (INIS)

    Kozikowska, Iwona; Binkowski, Łukasz J.; Szczepańska, Katarzyna; Sławska, Helena; Miszczuk, Katarzyna; Śliwińska, Magdalena; Łaciak, Tomasz; Stawarz, Robert

    2013-01-01

    Studies were conducted on samples taken from giving birth women (n = 40) living in Poland, representing three age groups: 19–25, 26–30 and 31–38 years old. Mercury concentrations were measured with CV-AAS in placenta, umbilical cord, cord blood and amniotic fluid. The placentas weight did not exceed the 750 g value and was heavier than 310 g. Mean values of Hg concentrations in blood, placenta and umbilical cord were similar (c.a. 9 μg/g). High levels of mercury were noted in cord blood which in 75% of all observations exceeded (up to 17 μg/L) the safe dose set by US EPA (5.8 μg/L). No statistically significant differences in medium level of Hg in all the studied tissues among age groups of women were observed. Positive correlations between Hg concentrations in placenta and umbilical cord and cord blood were revealed as well as some negative ones between mercury concentrations and pregnancy parameters. -- Highlights: •Concentrations of mercury in cord blood exceed the safety threshold level. •Maternal age was not an influential factor of Hg concentrations in studied samples. •Positive correlations between Hg levels in different tissues were observed. •Negative correlation between Hg concentrations and pregnancy parameters were noted. -- Maternal age was not an influential factor of mercury concentrations in studied samples. 75% of cord blood samples exceeded the Hg threshold concentration

  19. Dialkyl phosphates in amniotic fluid as a biomarker of fetal exposure to organophosphates in Crete, Greece; association with fetal growth.

    Science.gov (United States)

    Koutroulakis, D; Sifakis, S; Tzatzarakis, M N; Alegakis, A K; Theodoropoulou, E; Kavvalakis, M P; Kappou, D; Tsatsakis, A M

    2014-07-01

    The aim of this study was to evaluate fetal exposure to organophosphate pesticides (OPs) by measuring their non-specific dialkyl-phosphate metabolites (DAPs) in amniotic fluid (AF), and to examine the potential association between prenatal exposure and fetal growth. AF samples were collected from 415 women during the second gestational trimester. The determined OPs metabolites were DMP, DMTP, DEP, DETP, and DEDTP. DAPs were extracted by liquid-solid extraction, derivatized and analyzed by gas chromatography-mass spectrometry. 97.8% of AF samples were positive for at least one DAP. DAPs levels did not differ between urban and rural areas. Macrosomic neonates have significantly higher sum levels of DMPs (p=0.043), which exerted a linear positive association with birth-weight centile (b=4.43, p=0.016). Conclusively, as DAPs are detectable in AF they may be used as a potential biomarker of fetal exposure to OPs. Sum levels of DMPs appear to be associated with birth weight independently of other covariates. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Preterm prelabor rupture of membranes (PPROM) is not associated with presence of viral genomes in the amniotic fluid.

    Science.gov (United States)

    Bopegamage, Shubhada; Kacerovsky, Marian; Tambor, Vojtech; Musilova, Ivana; Sarmirova, Sona; Snelders, Eveline; de Jong, Arjan S; Vari, Sandor G; Melchers, Willem J G; Galama, Jochem M D

    2013-11-01

    The role of viral infections in preterm prelabor rupture of the membranes (PPROM) is not established. Studies on the presence of viral genomes in the amniotic fluid (AF) collected in pregnancies complicated by PPROM show contradictory outcomes. To investigate AF samples of PPROM pregnancies for the presence of viral genomes. AF samples from patients with PPROM were collected during a 4-year (2008-2012) observational study. 174 women were included with selection criteria of singleton pregnancy, PPROM, and maternal age of 18 years and above. PCR was used for detection of human cytomegalovirus (HCMV), herpes simplex virus (HSV), parvovirus B19, human adenoviruses (HAdV), enteroviruses (EV) and human parechovirus (HPeV). The selection of these viral targets was based on literature regarding screening of AF for presence of viral genomes. Only a single sample was positive out of the 174 tested AFs, HCMV DNA was detected. PPROM is not associated with active viral infections. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Determination of bisphenol-A levels in human amniotic fluid samples by liquid chromatography coupled with mass spectrometry.

    Science.gov (United States)

    Chen, Mei; Edlow, Andrea G; Lin, Tao; Smith, Nicole A; McElrath, Thomas F; Lu, Chensheng

    2011-07-01

    Bisphenol A (BPA) is one of the environmental endocrine-disrupting chemicals used widely in common consumer products. There is an increasing concern about human exposure to BPA, particularly in fetuses, due to the potential adverse effects related to the estrogenic activity of BPA. In assessing environmental exposure to BPA, it is essential to have a sensitive, accurate, and specific analytical method, particularly for low BPA levels in complex sample matrices. In this study, we developed and validated an accurate, sensitive, and robust liquid chromatography-mass spectrometry (LC-MS) method for determining the BPA concentrations in human amniotic fluid (AF). In this method, BPA and the internal standards (13)C(12) -BPA were extracted from 500 μL of human AF using solid-phase extraction. Calibration curves were linear over a concentration range of 0.3-100 ng/mL for BPA. The analytes were quantitatively determined using LC-MS operated in a negative electrospray ionization selected ion monitoring mode. This validated method has been used successfully in the clinical sample analysis of BPA in second-trimester AF specimens. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Hyaluronic acid enhances proliferation of human amniotic mesenchymal stem cells through activation of Wnt/β-catenin signaling pathway

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Ru-Ming; Sun, Ren-Gang; Zhang, Ling-Tao; Zhang, Qing-Fang; Chen, Dai-Xiong [Guizhou Center for Translational Medicine, Affiliated Hospital of Zunyi Medical University, 149 Dalian Road, Zunyi 563000 (China); Zhong, Jian-Jiang, E-mail: jjzhong@sjtu.edu.cn [State Key Laboratory of Microbial Metabolism, and School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240 (China); Xiao, Jian-Hui, E-mail: jhxiao@yahoo.com [Guizhou Center for Translational Medicine, Affiliated Hospital of Zunyi Medical University, 149 Dalian Road, Zunyi 563000 (China)

    2016-07-15

    This study investigated the pro-proliferative effect of hyaluronic acid (HA) on human amniotic mesenchymal stem cells (hAMSCs) and the underlying mechanisms. Treatment with HA increased cell population growth in a dose- and time-dependent manner. Analyses by flow cytometry and immunocytochemistry revealed that HA did not change the cytophenotypes of hAMSCs. Additionally, the osteogenic, chondrogenic, and adipogenic differentiation capabilities of these hAMSCs were retained after HA treatment. Moreover, HA increased the mRNA expressions of wnt1, wnt3a, wnt8a, cyclin D1, Ki-67, and β-catenin as well as the protein level of β-catenin and cyclin D1 in hAMSCs; and the nuclear localization of β-catenin was also enhanced. Furthermore, the pro-proliferative effect of HA and up-regulated expression of Wnt/β-catenin pathway-associated proteins - wnt3a, β-catenin and cyclin D1 in hAMSCs were significantly inhibited upon pre-treatment with Wnt-C59, an inhibitor of the Wnt/β-catenin pathway. These results suggest that HA may positively regulate hAMSCs proliferation through regulation of the Wnt/β-catenin signaling pathway. - Highlights: • Hyaluronic acid (HA) could promote the proliferation of hAMSCs. • HA treatment dose not affect the pluripotency of hAMSCs. • HA increases hAMSCs proliferation through activation of Wnt/β-catenin signaling.

  3. Diagnosis of subclinical amniotic fluid infection prior to rescue cerclage using gram stain and glucose tests: an individual patient meta-analysis.

    Science.gov (United States)

    Lisonkova, Sarka; Sabr, Yasser; Joseph, K S

    2014-02-01

    Microbial invasion of the amniotic cavity (MIAC) can affect outcomes following rescue cerclage. We carried out a study to compare the diagnostic performance of the Gram stain and glucose tests for detecting subclinical MIAC. We used individual-level data from published studies on Gram stain, glucose, and amniotic fluid culture among women with preterm labour. We calculated the sensitivity, specificity, area under the curve (AUC) and other indices, with amniotic fluid culture results used as the gold standard. The probability of infection using both tests as predictors was also estimated using logistic regression. The rate of culture-confirmed MIAC was 11.8% (34 of 288 women). The Gram stain test yielded a sensitivity of 65% (95% CI 46% to 78%) and a specificity of 99% (95% CI 98% to 100%). A positive Gram stain or glucose test had a sensitivity of 88% (95% CI 72% to 96%) and a specificity of 87% (95% CI 82% to 90%), while a positive Gram stain and a positive glucose test had a sensitivity of 62% (95% CI 44% to 77%) and a specificity of 100% (95% CI 98% to 100%). The AUC for the tests were Gram stain 0.82 (95% CI 0.74 to 0.90), glucose 0.86 (95% CI 0.80 to 0.93), and combined Gram stain and glucose 0.92 (95% CI 0.86 to 0.98). Using the tests, singly or in combination, provided greater clinically important calibration, risk-stratification, and classification accuracy than using no tests. Amniotic fluid Gram stain and/or glucose testing provides substantially improved performance for the diagnosis of subclinical MIAC compared with no testing.

  4. Amniotic mesenchymal stem cells mitigate osteoarthritis progression in a synovial macrophage-mediated in vitro explant coculture model.

    Science.gov (United States)

    Topoluk, Natasha; Steckbeck, Kathleen; Siatkowski, Sandra; Burnikel, Brian; Tokish, John; Mercuri, Jeremy

    2017-11-13

    Osteoarthritis (OA) is a disease of the synovial joint marked by chronic, low-grade inflammation leading to cartilage destruction. Regenerative medicine strategies for mitigating OA progression and/or promoting cartilage regeneration must be assessed using models that mimic the hallmarks of OA. More specifically, these models should maintain synovial macrophage phenotype in their native micro-environment. Herein, an in vitro coculture model of patient-matched human OA cartilage and synovium was assessed for viability, macrophage phenotype, and progressive cartilage destruction in the presence of an inflammatory milieu. Additionally, the influence of synovial macrophages and their polarization within the model was defined using depletion studies. Finally, the model was used to compare the ability of human amniotic stem cells (hAMSCs) and human adipose stem cells (hADSCs) to mitigate OA progression. OA cocultures demonstrated progressive and significant reductions in chondrocyte viability and cartilage glycosaminoglycan content within a proinflammatory environment. Selective depletion of synovial macrophages resulted in significant decreases in M1:M2 percentage ratio yielding significant reductions in concentrations of interleukin-1 beta, matrix metalloproteinase-13 and attenuation of cartilage damage. Finally, hAMSCs were found to be more chondroprotective versus hADSCs as indicated by significantly improved OA chondrocyte viability (89.8 ± 2.4% vs. 58.4 ± 2.4%) and cartilage glycosaminoglycan content (499.0 ± 101.9 μg/mg dry weight vs. 155.0 ± 26.3 μg/mg dry weight) and were more effective at shifting OA synovial macrophage M1:M2 ratio (1.3:1 vs. 5:1), respectively. Taken together, the coculture model mimics salient features of OA, including macrophage-mediated cartilage destruction that was effectively abrogated by hAMSCs but not hADSCs. Copyright © 2017 John Wiley & Sons, Ltd.

  5. Cell-free DNA fragmentation patterns in amniotic fluid identify genetic abnormalities and changes due to storage.

    Science.gov (United States)

    Peter, Inga; Tighiouart, Hocine; Lapaire, Olav; Johnson, Kirby L; Bianchi, Diana W; Terrin, Norma

    2008-09-01

    Circulating cell-free DNA (cfDNA) has become a promising biomarker in prenatal diagnosis. However, despite extensive studies in different body fluids, cfDNA predictive value is uncertain owing to the confounding factors that can affect its levels, such as gestational age, maternal weight, smoking status, and medications. Residual fresh and archived amniotic fluid (AF) supernatants were obtained from gravid women (mean gestational age 17 wk) carrying euploid (N=36) and aneuploid (N=29) fetuses, to characterize cfDNA-fragmentation patterns with regard to aneuploidy and storage time (-80 degrees C). AF cfDNA was characterized by the real-time quantitative polymerase chain reaction amplification of glyceraldehyde-3-phosphate dehydrogenase, gel electrophoresis, and pattern recognition of the DNA fragmentation. The distributions of cfDNA fragment lengths were compared using 6 measures that defined the locations and slopes for the first and last peaks, after elimination of the confounding variables. This method allowed for the unique classification of euploid and aneuploid cfDNA samples in AF, which had been matched for storage time. In addition, we showed that archived euploid AF samples gradually lose long cfDNA fragments: this loss accurately distinguishes them from the fresh samples. We present preliminary data using cfDNA-fragmentation patterns, to uniquely distinguish between AF samples of pregnant women with regard to aneuploidy and storage time, independent of gestational age and initial DNA amount. In addition to potential applications in prenatal diagnosis, these data suggest that archived AF samples consist of large amounts of short cfDNA fragments, which are undetectable using standard real-time polymerase chain reaction amplification.

  6. Mesenchymal stem cells in synovial fluid increase after meniscus injury.

    Science.gov (United States)

    Matsukura, Yu; Muneta, Takeshi; Tsuji, Kunikazu; Koga, Hideyuki; Sekiya, Ichiro

    2014-05-01

    Although relatively uncommon, spontaneous healing from a meniscus injury has been observed even within the avascular area. This may be the result of the existence of mesenchymal stem cells in synovial fluid. The purpose of this study was to investigate whether mesenchymal stem cells existed in the synovial fluid of the knee after meniscus injury. Synovial fluid was obtained from the knees of 22 patients with meniscus injury just before meniscus surgery and from 8 volunteers who had no history of knee injury. The cellular fraction of the synovial fluid was cultured for 14 days followed by analysis for multilineage potential and presentation of surface antigens characteristic of mesenchymal stem cells. Colony-forming efficiency and proliferation potential were also compared between the two groups. Cells with characteristics of mesenchymal stem cells were observed in the synovial fluid of injured knees to a much greater degree than in uninjured knees. The colony-forming cells derived from the synovial fluid of the knee with meniscus injury had multipotentiality and surface epitopes identical to mesenchymal stem cells. The average number of colony formation, obtained from 1 mL of synovial fluid, in meniscus-injured knees was 250, higher than that from healthy volunteers, which was 0.5 (p < 0.001). Total colony number per synovial fluid volume was positively correlated with the postinjury period (r = 0.77, p < 0.001). Mesenchymal stem cells were found to exist in synovial fluid from knees after meniscus injury. Mesenchymal stem cells were present in higher numbers in synovial fluid with meniscus injury than in normal knees. Total colony number per synovial fluid volume was positively correlated with the postinjury period. Our current human study and previous animal studies suggest the possibility that mesenchymal stem cells in synovial fluid increase after meniscus injury contributing to spontaneous meniscus healing.

  7. Karyotype analysis with amniotic fluid in 12365 pregnant women with indications for genetic amniocentesis and strategies of prenatal diagnosis.

    Science.gov (United States)

    Xiao, H; Yang, Y L; Zhang, C Y; Liao, E J; Zhao, H R; Liao, S X

    2016-01-01

    We explored the strategies of prenatal diagnosis by foetal karyotype analysis in pregnant women with indications for genetic amniocentesis. Karyotype analysis of amniotic fluid was performed on 12365 pregnant women with indications for genetic amniocentesis. The detection rates and distributions of abnormal karyotypes were observed in a variety of indications for genetic amniocentesis. The detection rates of abnormal karyotype were 57.4% in either a mother or father with chromosomal abnormality, 8.5% in the pregnant women with pathological ultrasound finding (PUF), 2.79% in the pregnant women with advanced age (35 years and over) and 2.23% in the women with abnormal maternal serum screening (MSS) tests. Foetal abnormal karyotype was found in 86 pregnant women with PUF; of the 86 pregnant women, 42 had trisomy 13, 18 or 21. Of the 12365 pregnant women, foetal abnormal karyotype was found in 428 (3.46%); of the 428 foetuses, only 154 had trisomy 13, 18 or 21. In the pregnant women with abnormal MSS, 111 foetuses had abnormal karyotype, but only 36 foetuses had trisomy 13, 18 or 21. We conclude that (1) ultrasound is an important approach to prevent the birth of foetuses with chromosomal disease. (2) Non-invasive prenatal DNA detection cannot completely replace invasive prenatal diagnosis and MSS. (3) The strategies of prenatal diagnosis: Genetic amniocentesis is strongly recommended for the pregnant women with indications for genetic amniocentesis. For pregnant women who refuse invasive prenatal diagnosis, non-invasive prenatal DNA detection is first performed. If the results of non-invasive prenatal DNA detection are negative, the pregnant women are followed up by ultrasound; if the results of non-invasive prenatal DNA detection are positive, the pregnant women should undergo invasive prenatal diagnosis.

  8. Transvaginal cervical length and amniotic fluid index: can it predict delivery latency following preterm premature rupture of membranes?

    Science.gov (United States)

    Mehra, Suwan; Amon, Erol; Hopkins, Sarah; Gavard, Jeffrey A; Shyken, Jaye

    2015-03-01

    We sought to determine whether transvaginal cervical length (TVCL), amniotic fluid index (AFI), or a combination of both can predict delivery latency within 7 days in women presenting with preterm premature rupture of membranes (PPROM). This was a prospective observational study of TVCL measurements in 106 singleton pregnancies with PPROM between 23-33 weeks. Delivery latency was defined as the period (in days) from the initial TVCL after PPROM to delivery of the infant, with our primary outcome being delivery within 7 days of TVCL. The independent predictability of significant characteristics for delivery within 7 days was determined using multiple logistic regression. Sensitivity, specificity, and predictive values were used to examine whether the presence of a short TVCL, AFI, or a combination of both affected the risk of delivery within 7 days. Delivery within 7 days occurred in 51/106 (48%) of pregnancies. Median duration (interquartile range) from PPROM to delivery and TVCL to delivery was 8 days (4.0-16.0) and 8 days (3.0-15.0), respectively. Using multiple regression TVCL as a continuous variable (odds ratio, 0.65; 95% confidence interval, 0.44-0.97; P 7 days for TVCL >2 cm alone was 61%. This predictive value changed when analyzed in conjunction with an AFI ≤5 cm and >5 cm at 42% and 89%, respectively. A shorter TVCL and an AFI ≤5 cm independently predict delivery within 7 days in women presenting with PPROM. The combination of an AFI >5 cm and TVCL >2 cm greatly improved the potential to remain undelivered at 7 days following cervical length assessment. These findings may be helpful for counseling and optimizing maternal and neonatal care in women with PPROM. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Modulation of amniotic fluid activin-a and inhibin-a in women with preterm premature rupture of the membranes and infection-induced preterm birth.

    Science.gov (United States)

    Rosenberg, Victor A; Buhimschi, Irina A; Dulay, Antonette T; Abdel-Razeq, Sonya S; Oliver, Emily A; Duzyj, Christina M; Lipkind, Heather; Pettker, Christian M; Buhimschi, Catalin S

    2012-02-01

    Activins and inhibins are important modulators of inflammatory processes. We explored activation of amniotic fluid (AF) activin-A and inhibin-A system in women with intra-amniotic infection and preterm premature rupture of the membranes (PPROM). We analyzed 78 AF samples: '2nd trimester-control' (n=12), '3rd trimester-control' (n=14), preterm labor with intact membranes [positive-AF-cultures (n=13), negative-AF-cultures (n=13)], and PPROM [positive-AF-cultures (n=13), negative-AF-cultures (n=13)]. Activin-A levels were evaluated ex-vivo following incubation of amniochorion and placental villous explants with Gram-negative lipopolysaccharide (LPS) or Gram-positive (Pam3Cys) bacterial mimics. Ability of recombinant activin-A and inhibin-A to modulate inflammatory reactions in fetal membranes was explored through explants' IL-8 release. Activin-A and inhibin-A were present in human AF and were gestational age-regulated. Activin-A was significantly upregulated by infection. Lower inhibin-A levels were seen in PPROM. LPS elicited release of activin-A from amniochorion, but not from villous explants. Recombinant activin-A stimulated IL-8 release from amniochorion, an effect that was not reversed by inhibin-A. Human AF activin-A and inhibin-A are involved in biological processes linked to intra-amniotic infection/inflammation-induced preterm birth. © 2011 John Wiley & Sons A/S.

  10. LC-MS/MS multiplex analysis of lysosphingolipids in plasma and amniotic fluid: A novel tool for the screening of sphingolipidoses and Niemann-Pick type C disease.

    Directory of Open Access Journals (Sweden)

    Magali Pettazzoni

    Full Text Available The biological diagnosis of sphingolipidoses currently relies on the measurement of specific enzymatic activities and/or genetic studies. Lysosphingolipids have recently emerged as potential biomarkers of sphingolipidoses and Niemann-Pick type C in plasma.We developed a sensitive and specific method enabling the simultaneous quantification of lysosphingolipids by LC-MS/MS: lysoglobotriaosylceramide for Fabry disease, lysohexosylceramide (i.e. lysoglucosylceramide and/or lysogalactosylceramide for Gaucher and Krabbe diseases, lysosphingomyelin and its carboxylated analogue lysosphingomyelin-509 for Niemann-Pick type A or B, and C diseases, lysoGM1 ganglioside for GM1gangliosidosis and lysoGM2 ganglioside for GM2 gangliosidosis.The diagnostic performances were validated in plasma samples analysing a large series of patients affected with sphingolipidoses and Niemann-Pick type C disease (n = 98, other inborn errors of metabolism (n = 23, and controls (n = 228. The multiplex measurement of lysosphingolipids allowed the screening of Fabry (including female patients and late-onset variants, Gaucher and infantile Krabbe, Niemann-Pick type A/B and C diseases with high sensitivity and specificity. LysoGM1 and LysoGM2 were elevated in most of the patients affected with GM1 and GM2 gangliosidosis respectively. In amniotic fluid supernatant from pregnancies presenting non-immune hydrops fetalis (n = 77, including previously diagnosed Gaucher (n = 5, GM1 gangliosidosis (n = 4 and galactosialidosis (n = 4 fetuses and from normal pregnancies (n = 15, a specific and dramatic increase of lysohexosylceramide was observed only in the Gaucher amniotic fluid samples.This multiplex assay which allows the simultaneous measurement of lysosphingolipids in plasma modifies the diagnostic strategy of sphingolipidoses and Niemann-Pick type C. Furthermore, in pregnancies presenting non-immune hydrops fetalis, lysohexosylceramide measurement in amniotic fluid offers a rapid

  11. High performance liquid chromatography-tandem mass spectrometric assay of dexmedetomidine in plasma, urine and amniotic fluid samples for pregnant ewe model.

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    Cui, Z; Chow, D S-L; Wu, L; Lazar, D A; Rodrigo, R; Olutoye, O O; Olutoye, O A

    2014-06-15

    Dexmedetomidine (DEX; Precedex(®)), approved by the Food and Drug Administration (FDA) in 1999 as a sedative for use in the intensive care unit, is a potent and highly selective α2-adrenoceptor agonist with significant sedative, analgesic and anxiolytic effects. However, the research of DEX use during pregnancy is limited and the impact of DEX on the fetal development is unclear. This article describes a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) assay suitable for various biomatrices of plasma, urine and amniotic fluid, as a prerequisite for pharmacokinetic characterization of DEX in the pregnant ewe model. DEX and testosterone (internal standard; IS) were extracted from 200μL of plasma, urine or amniotic fluid with ethyl acetate. The HPLC resolution was achieved on an Agilent ZORBAX SB-CN column with a gradient elution at a flow rate of 0.5mL/min using a mobile phase of 5-100% of acetonitrile with 0.5% formic acid (mobile phase B) in water (mobile phase A). The detection was performed by a triple quadrupole tandem mass spectrometer with positive electrospray ionization. The precursor/product transitions (m/z) in the positive ion mode [M+H](+) were m/z 201.5→95.4 for DEX and m/z 289.2→109.1 for IS. The method was validated in the concentration range of 25 (lower limit of quantification; LLOQ)-5000pg/mL for both maternal and fetal plasma, and of 50 (LLOQ)-5000pg/mL for urine and amniotic fluid, respectively. The intra- and inter-day precision and accuracy were within ±9%. The overall recoveries of DEX were 82.9-87.2%, 85.7-88.4%, 86.2-89.7% and 83.7-88.1% for maternal plasma, urine, fetal plasma and amniotic fluid, respectively. The percentage matrix factors in different biomatrices were less than 120%. Stability studies demonstrated that DEX was stable after three freeze/thaw cycles, in the autosampler tray at 20°C for 24h and during the 3h sample preparation at room temperature. The validated HPLC-MS/MS method has been

  12. Evaluation of the Cepheid Xpert GBS Assay for Rapid Detection of Group B Streptococci in Amniotic Fluids from Pregnant Women with Premature Rupture of Membranes

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    Bourgeois-Nicolaos, Nadege; Cordier, Anne-Gael; Guillet-Caruba, Christelle; Casanova, François; Benachi, Alexandra

    2013-01-01

    The Xpert GBS real-time PCR assay for the detection of group B streptococci (GBS) in antepartum screening samples was evaluated on amniotic fluid samples collected from 139 women with premature rupture of membrane at term. When any intrapartum positive result from the Xpert GBS or culture was considered a true positive, the sensitivities of the Xpert GBS and culture were 92.3% and 84.6%, respectively. This assay could enhance exact identification of candidates for intrapartum antibiotic prophylaxis. PMID:23390279

  13. Provision of Amniotic Fluid During Parenteral Nutrition Increases Weight Gain With Limited Effects on Gut Structure, Function, Immunity, and Microbiology in Newborn Preterm Pigs

    DEFF Research Database (Denmark)

    Østergaard, Mette Viberg; Liang Shen, Rene; Støy, Ann Cathrine Findal

    2016-01-01

    Background: Small enteral boluses with human milk may reduce the risk of subsequent feeding intolerance and necrotizing enterocolitis in preterm infants receiving parenteral nutrition (PN). We hypothesized that feeding amniotic fluid, the natural enteral diet of the mammalian fetus, will have...... and IL8 were increased in AF pigs, while blood biochemistry and amino acid levels were minimally affected. Conclusion: Enteral boluses of AF were well tolerated in the first 5 days of life in preterm pigs receiving PN. Enteral provision of AF before the initiation of milk feeding may stimulate body...

  14. Transplanted human amniotic membrane-derived mesenchymal stem cells ameliorate carbon tetrachloride-induced liver cirrhosis in mouse.

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    DingGuo Zhang

    Full Text Available BACKGROUND: Human amniotic membrane-derived mesenchymal stem cells (hAMCs have the potential to reduce heart and lung fibrosis, but whether could reduce liver fibrosis remains largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: Hepatic cirrhosis model was established by infusion of CCl₄ (1 ml/kg body weight twice a week for 8 weeks in immunocompetent C57Bl/6J mice. hAMCs, isolated from term delivered placenta, were infused into the spleen at 4 weeks after mice were challenged with CCl₄. Control mice received only saline infusion. Animals were sacrificed at 4 weeks post-transplantation. Blood analysis was performed to evaluate alanine aminotransferase (ALT and aspartate aminotransferase (AST. Histological analysis of the livers for fibrosis, hepatic stellate cells activation, hepatocyte apoptosis, proliferation and senescence were performed. The donor cell engraftment was assessed using immunofluorescence and polymerase chain reaction. The areas of hepatic fibrosis were reduced (6.2%±2.1 vs. control 9.6%±1.7, p<0.05 and liver function parameters (ALT 539.6±545.1 U/dl, AST 589.7±342.8 U/dl,vs. control ALT 139.1±138.3 U/dl, p<0.05 and AST 212.3±110.7 U/dl, p<0.01 were markedly ameliorated in the hAMCs group compared to control group. The transplantation of hAMCs into liver-fibrotic mice suppressed activation of hepatic stellate cells, decreased hepatocyte apoptosis and promoted liver regeneration. More interesting, hepatocyte senescence was depressed significantly in hAMCs group compared to control group. Immunofluorescence and polymerase chain reaction revealed that hAMCs engraftment into host livers and expressed the hepatocyte-specific markers, human albumin and α-fetoproteinran. CONCLUSIONS/SIGNIFICANCE: The transplantation of hAMCs significantly decreased the fibrosis formation and progression of CCl₄-induced cirrhosis, providing a new approach for the treatment of fibrotic liver disease.

  15. Combined HLA matched limbal stem cells allograft with amniotic membrane transplantation as a prophylactic surgical procedure to prevent corneal graft rejection after penetrating keratoplasty: case report

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    Paolo Capozzi

    2014-09-01

    Full Text Available Purpose. To determine if the use of combined HLA matched limbal stem cells allograft with amniotic membrane transplantation (AMT is a safe and effective prophylactic surgical procedure to prevent corneal graft after penetrating keratoplasty (PK. Methods. We report the case of a 17 years old patient with a history of congenital glaucoma, trabeculectomy and multiple corneal graft rejections, presenting total limbal cell deficiency. To reduce the possibility of graft rejection in the left eye after a new PK, a two step procedure was performed. At first the patient underwent a combined HLA matched limbal stem cells allograft (LAT and AMT and then, 10 months later, a new PK. Results. During 12 months of follow-up, the corneal graft remained stable and smooth, with no sign of graft rejection. Conclusions. In our patient, the prophylactic use of LAT from HLA-matched donors and AMT before PK, may result in a better prognosis of corneal graft survival.

  16. Amniotic Fluid Analysis

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  17. Amniotic Mesenchymal Stem Cells: A New Source for Hepatocyte-Like Cells and Induction of CFTR Expression by Coculture with Cystic Fibrosis Airway Epithelial Cells

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    Valentina Paracchini

    2012-01-01

    Full Text Available Cystic fibrosis (CF is a monogenic disease caused by mutations in the CF transmembrane conductance regulator (CFTR gene, with lung and liver manifestations. Because of pitfalls of gene therapy, novel approaches for reconstitution of the airway epithelium and CFTR expression should be explored. In the present study, human amniotic mesenchymal stem cells (hAMSCs were isolated from term placentas and characterized for expression of phenotypic and pluripotency markers, and for differentiation potential towards mesoderm (osteogenic and adipogenic lineages. Moreover, hAMSCs were induced to differentiate into hepatocyte-like cells, as demonstrated by mixed function oxidase activity and expression of albumin, alpha1-antitrypsin, and CK19. We also investigated the CFTR expression in hAMSCs upon isolation and in coculture with CF airway epithelial cells. Freshly isolated hAMSCs displayed low levels of CFTR mRNA, which even decreased with culture passages. Following staining with the vital dye CM-DiI, hAMSCs were mixed with CFBE41o- respiratory epithelial cells and seeded onto permeable filters. Flow cytometry demonstrated that 33–50% of hAMSCs acquired a detectable CFTR expression on the apical membrane, a result confirmed by confocal microscopy. Our data show that amniotic MSCs have the potential to differentiate into epithelial cells of organs relevant in CF pathogenesis and may contribute to partial correction of the CF phenotype.

  18. Comparison of placental alpha microglobulin-1 in vaginal fluid with intra-amniotic injection of indigo carmine for the diagnosis of rupture of membranes.

    Science.gov (United States)

    Sosa, Claudio G; Herrera, Edward; Restrepo, Juan C; Strauss, Alexander; Alonso, Justo

    2014-09-01

    The purpose of this study was to compare the diagnostic performance of an immunoassay for placental alpha microglobulin-1 (PAMG-1) in vaginal fluid with the intra-amniotic instillation of indigo carmine for the diagnosis of membrane rupture in patients with an equivocal status. A prospective multi-site study was performed involving women reporting signs, symptoms, or complaints suggestive of rupture of membrane (ROM) between 21 and 42 weeks of gestation without obvious leakage of fluid from the cervical os during sterile speculum examination and without confirmation of ROM by traditional methods. A total of 140 patients were recruited with a prevalence of ROM of 19.3%. The PAMG-1 test had a sensitivity of 100.0% [confidence interval (CI) 0.87-1.0], specificity of 99.1% [(CI) 0.95-0.99], positive predictive value of 96.3% [(CI) 0.82-0.99], negative predictive value of 100.0% [(CI) 0.97-1.0], and ± likelihood ratios of 74.6 [(CI) 20.31-274.51] and 0.0 [(CI) 0.00-0.98]. The PAMG-1 immunoassay in vaginal fluid yielded results that were comparable to those of the instillation of indigo carmine into the amniotic cavity; therefore, we propose that PAMG-1 is a sensitive and specific test to assess ROM in patients with an equivocal diagnosis based on simple tests. This finding prompts consideration for the use of the noninvasive PAMG-1 test in situations where the use of the invasive dye test is not practical.

  19. Gender-Typed Play and Amniotic Testosterone

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    Knickmeyer, Rebecca Christine; Wheelwright, Sally; Taylor, Kevin; Raggatt, Peter; Hackett, Gerald; Baron-Cohen, Simon

    2005-01-01

    Sex differences in play are apparent in a number of mammalian species, including humans. Prenatal testosterone may contribute to these differences. The authors report the first attempt to correlate gender-typed play in a normative sample of humans with measurements of amniotic testosterone (aT). Testosterone was measured in the amniotic fluid of…

  20. Term amniotic membrane is a high throughput source for multipotent mesenchymal stem cells with the ability to differentiate into endothelial cells in vitro

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    Pasquinelli Gianandrea

    2007-02-01

    Full Text Available Abstract Background Term Amniotic membrane (AM is a very attractive source of Mesenchymal Stem Cells (MSCs due to the fact that this fetal tissue is usually discarded without ethical conflicts, leading to high efficiency in MSC recovery with no intrusive procedures. Here we confirmed that term AM, as previously reported in the literature, is an abundant source of hMSCs; in particular we further investigated the AM differentiation potential by assessing whether these cells may also be committed to the angiogenic fate. In agreement with the recommendation of the International Society for Cellular Therapy, the mesenchymal cells herein investigated were named Amniotic Membrane-human Mesenchymal Stromal Cells (AM-hMSC. Results The recovery of hMSCs and their in vitro expansion potential were greater in amniotic membrane than in bone marrow stroma. At flow cytometry analysis AM-hMSCs showed an immunophenotypical profile, i.e., positive for CD105, CD73, CD29, CD44, CD166 and negative for CD14, CD34, CD45, consistent with that reported for bone marrow-derived MSCs. In addition, amniotic membrane-isolated cells underwent in vitro osteogenic (von Kossa stain, adipogenic (Oil Red-O stain, chondrogenic (collagen type II immunohistochemichal detection and myogenic (RT-PCR MyoD and Myogenin expression as well as desmin immunohistochemical detection differentiation. In angiogenic experiments, a spontaneous differentiation into endothelial cells was detected by in vitro matrigel assay and this behaviour has been enhanced through Vascular Endothelial Growth Factor (VEGF induction. According to these findings, VEGF receptor 1 and 2 (FLT-1 and KDR were basally expressed in AM-hMSCs and the expression of endothelial-specific markers like FLT-1 KDR, ICAM-1 increased after exposure to VEGF together with the occurrence of CD34 and von Willebrand Factor positive cells. Conclusion The current study suggests that AM-hMSCs may emerge as a remarkable tool for the cell

  1. Relationships between sex hormones assessed in amniotic fluid, and maternal and umbilical cord serum: what is the best source of information to investigate the effects of fetal hormonal exposure?

    Science.gov (United States)

    van de Beek, Cornelieke; Thijssen, Jos H H; Cohen-Kettenis, Peggy T; van Goozen, Stephanie H M; Buitelaar, Jan K

    2004-12-01

    Levels of testosterone (T) (total and free), androstenedione (A4), dehydroepiandrosterone sulphate (DHEAS), sex hormone-binding globulin (SHBG), and estradiol (E2) were measured by radioimmunoassay (RIA) in 156 normal pregnancies (77 male and 79 female fetuses). Samples were obtained from amniotic fluid, 2nd and 3rd trimester maternal serum, and umbilical cord serum at birth. During the critical period of brain differentiation, at the beginning of the second trimester of pregnancy, sex differences in T and A4 were found in amniotic fluid and not in maternal serum. This finding adds to the fact that mostly low and nonsignificant correlations were found for the different androgenic hormones between levels assessed in amniotic fluid and maternal plasma at this particular and very sensitive period of fetal brain development. On the other hand, high correlations were found for the same hormones between the samples of maternal serum in the 2nd and the 3rd trimester. Our data show that, of all available sources, amniotic fluid seems to be the best candidate to investigate the effects of early fetal androgen exposure.

  2. An Update on Maternal Hydration Strategies for Amniotic Fluid Improvement in Isolated Oligohydramnios and Normohydramnios: Evidence from a Systematic Review of Literature and Meta-Analysis.

    Science.gov (United States)

    Gizzo, Salvatore; Noventa, Marco; Vitagliano, Amerigo; Dall'Asta, Andrea; D'Antona, Donato; Aldrich, Clive J; Quaranta, Michela; Frusca, Tiziana; Patrelli, Tito Silvio

    2015-01-01

    Several trials aimed at evaluating the efficacy of maternal hydration (MH) in increasing amniotic-fluid-volume (AFV) in pregnancies with isolated oligohydramnios or normohydramnos have been conducted. Unfortunately, no evidences support this intervention in routine-clinical-practice. The aim of this systematic-literature-review and meta-analysis was to collect all data regarding proposed strategies and their efficacy in relation to each clinical condition for which MH-therapy was performed with the aim of increasing amniotic-fluid (AF) and improving perinatal outcomes. A systematic literature search was conducted in electronic-database MEDLINE, EMBASE, ScienceDirect and the Cochrane-Library in the time interval between 1991 and 2014. Following the identification of eligible trials, we estimated the methodological quality of each study (using QADAS-2) and clustered patients according to the following outcome measures: route of administration (oral versus intravenous versus combined), total daily dose of fluids administered (2000), duration of hydration therapy: (1 day, >1 day but 1 week), type of fluid administered (isotonic versus hypotonic versus combination). In isolated-oligohydramnios (IO), maternal oral hydration is more effective than intravenous hydration and hypotonic solutions superior to isotonic solutions. The improvement in AFV appears to be time-dependent rather than daily-dose dependent. Regarding normohydramnios pregnancies, all strategies seem equivalent though the administration of hypotonic-fluid appears to have a slightly greater effect than isotonic-fluid. Regarding perinatal outcomes, data is fragmentary and heterogeneous and does not allow us to define the real clinical utility of MH. Available data suggests that MH may be a safe, well-tolerated and useful strategy to improve AFV especially in cases of IO. In view of the numerous obstetric situations in which a reduced AFV may pose a threat, particularly to the fetus, the possibility of

  3. Secretome profiling of cytokines and growth factors reveals that neuro-glial differentiation is associated with the down-regulation of Chemokine Ligand 2 (MCP-1/CCL2) in amniotic fluid derived-mesenchymal progenitor cells.

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    Miceli, Marco; Dell'Aversana, Carmela; Russo, Rosita; Rega, Camilla; Cupelli, Lorenzo; Ruvo, Menotti; Altucci, Lucia; Chambery, Angela

    2016-02-01

    Secreted cytokines and growth factors play a key role in the modulation of stem cell proliferation, differentiation and survival. To investigate the interplay between the changes in their expression levels, we used the newly characterized human amniotic fluid derived-mesenchymal progenitor MePR-2B cell line differentiated to a neuro-glial phenotype and exploited the very high sensitivity and versatility of magnetic beads-based immunoassays. We found that a sub-set of proteins, including the cytokines IL-6, TNFα, IL-15, IFNγ, IL-8, IL-1ra, MCP-1/CCL2, RANTES and the growth factor PDGFbb, underwent a significant down-regulation following neuro-glial differentiation, whereas the expression levels of IL-12 p70, IL-5, IL-7, bFGF, VEGF and G-CSF were increased. The role of MCP-1/CCL2, previously identified as a regulator of neural progenitor stem cell differentiation, has been further investigated at transcriptional level, revealing that both the chemokine and its receptor are co-expressed in MePR-2B cells and that are regulated upon differentiation, suggesting the presence of an autocrine and paracrine loop in differentiating cells. Moreover, we demonstrated that exogenous CCL2 is capable to affect neuro-glial differentiation in MePR-2B cells, thus providing novel evidences for the potential involvement of chemokine-mediated signaling in progenitor/stem cells differentiation processes and fate specification. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Isolation and culture exploration of Anas platyrhynchos amniotic fluid stem cells in vitro

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    Mingming Ning

    2017-06-01

    Conclusion: DAFSCs can be isolated from matrix that have strong self-renewal capacity in vitro. DAFSCs can be induced into adipocyte in vitro. These testify that DAFSCs can be an ideal seeded cells having potentials for preservation and utilization of rare genetic resources. [J Adv Vet Anim Res 2017; 4(2.000: 140-146

  5. Term Amniotic membrane is a high throughput source for multipotent Mesenchymal Stem Cells with the ability to differentiate into endothelial cells in vitro

    DEFF Research Database (Denmark)

    Alviano, Francesco; Fossati, Valentina; Marchionni, Cosetta

    2007-01-01

    BACKGROUND: Term Amniotic membrane (AM) is a very attractive source of Mesenchymal Stem Cells (MSCs) due to the fact that this fetal tissue is usually discarded without ethical conflicts, leading to high efficiency in MSC recovery with no intrusive procedures. Here we confirmed that term AM...... II immunohistochemichal detection) and myogenic (RT-PCR MyoD and Myogenin expression as well as desmin immunohistochemical detection) differentiation. In angiogenic experiments, a spontaneous differentiation into endothelial cells was detected by in vitro matrigel assay and this behaviour has been...... enhanced through Vascular Endothelial Growth Factor (VEGF) induction. According to these findings, VEGF receptor 1 and 2 (FLT-1 and KDR) were basally expressed in AM-hMSCs and the expression of endothelial-specific markers like FLT-1 KDR, ICAM-1 increased after exposure to VEGF together with the occurrence...

  6. Zika virus detected in amniotic fluid and umbilical cord blood in an in vitro fertilization-conceived pregnancy in Venezuela.

    Science.gov (United States)

    Benjamin, Isaac; Fernández, Gissel; Figueira, José Valentin; Parpacén, Leticia; Urbina, María Teresa; Medina, Randolfo

    2017-06-01

    To describe the consequences of Zika virus infection at 10 weeks of gestation in an IVF-conceived pregnancy in Venezuela. A case report. Private assisted reproduction unit. A 36-year-old patient who conceived her first pregnancy through IVF and became infected with Zika virus at 10 weeks' gestation in Venezuela. In vitro fertilization with fresh ET. Clinical, laboratory, and imaging Zika diagnostic methods. Zika virus detection by real-time polymerase chain reaction (PCR) in maternal plasma, PCR in amniotic fluid and umbilical cord blood. Ultrasonography findings of anatomic abnormalities. Zika infection was confirmed at 10 weeks' gestation by real-time PCR; ultrasound results appeared normal. At 19 weeks' gestation, an ultrasound revealed biometry on three SDs below the means for all parameters but with no apparent anatomic abnormality. Zika virus was positive in maternal urine and amniotic fluid by PCR at 19 weeks' gestation. Ultrasound at 21 weeks + 4 days of gestation showed fetal cerebellar hypoplasia with ventricular dysmorphia, particularly marked on the left, consistent with microcephaly and ventriculomegaly. Because of the poor prognosis, pregnancy was interrupted at 24 weeks' gestation, in France. The PCR in umbilical cord blood taken in this procedure was positive for Zika virus. Initial ultrasound findings in pregnancy may not be informative. Only at 21 weeks + 4 days of gestation did an ultrasound reveal fetal microcephaly and ventriculomegaly. Combined clinical, laboratory, and imaging findings provided a complete picture of the severe damage caused by Zika infection. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  7. Determination of 2',3'-dideoxycytidine in maternal plasma, amniotic fluid, placental and fetal tissues by high-performance liquid chromatography.

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    Ding, Yan; Williamson, Leah N; White, Catherine A; Bartlett, Michael G

    2004-11-25

    2',3'-Dideoxycytidine (DDC) is a nucleoside reverse transcriptase inhibitor that has been shown to inhibit the human immunodeficiency virus (HIV). DDC is a candidate for treatment of pregnant women to prevent prenatal transmission of HIV/AIDS to their unborn children. A quick and simple high-performance liquid chromatography (HPLC) method has been developed and validated for the determination of DDC concentrations in samples collected from a pregnant rat model (maternal plasma, amniotic fluid, placental and fetal tissues). Extraction of DDC and its internal standard 2',3'-dideoxy-3'-thiacytidine (3TC) in plasma and amniotic fluid was carried out by protein precipitation. Extraction from placental and fetal homogenates was achieved by solid phase extraction using Waters Oasis HLB solid phase extraction cartridges. Chromatographic separation was achieved on a Waters Spherisorb S3W silica column (4.6 mm x 100 mm) equipped with a Phenomenex guard column. The mobile phase used was 10% methanol in water with 22 mM formic acid. The flow rate was 0.5 ml/min, and the detection wavelength was optimized at 275 nm. Under these chromatographic conditions, DDC eluted around 12 min, and 3TC eluted around 10 min. The calibration curves for each day of validation and analysis showed good linear response through the range of 0.15-75.0 microg/ml in each of the four matrices. The relative recovery for DDC in each of the matrices ranged from 87.8% to 103.0%. Acceptable intra- and inter-day assay precision (<15% R.S.D.) and accuracy (<15% error) were observed over 0.15-75.0 microg/ml for all four matrices.

  8. Maternal intravenous treatment with either azithromycin or solithromycin clears Ureaplasma parvum from the amniotic fluid in an ovine model of intrauterine infection.

    Science.gov (United States)

    Miura, Yuichiro; Payne, Matthew S; Keelan, Jeffrey A; Noe, Andres; Carter, Sean; Watts, Rory; Spiller, Owen B; Jobe, Alan H; Kallapur, Suhas G; Saito, Masatoshi; Stock, Sarah J; Newnham, John P; Kemp, Matthew W

    2014-09-01

    Intrauterine infection with Ureaplasma spp. is strongly associated with preterm birth and adverse neonatal outcomes. We assessed whether combined intraamniotic (IA) and maternal intravenous (IV) treatment with one of two candidate antibiotics, azithromycin (AZ) or solithromycin (SOLI), would eradicate intrauterine Ureaplasma parvum infection in a sheep model of pregnancy. Sheep with singleton pregnancies received an IA injection of U. parvum serovar 3 at 85 days of gestational age (GA). At 120 days of GA, animals (n=5 to 8/group) received one of the following treatments: (i) maternal IV SOLI with a single IA injection of vehicle (IV SOLI only); (ii) maternal IV SOLI with a single IA injection of SOLI (IV+IA SOLI); (iii) maternal IV AZ and a single IA injection of vehicle (IV AZ only); (iv) maternal IV AZ and a single IA injection of AZ (IV+IA AZ); or (v) maternal IV and single IA injection of vehicle (control). Lambs were surgically delivered at 125 days of GA. Treatment efficacies were assessed by U. parvum culture, quantitative PCR, enzyme-linked immunosorbent assay, and histopathology. Amniotic fluid (AF) from all control animals contained culturable U. parvum. AF, lung, and chorioamnion from all AZ- or SOLI-treated animals (IV only or IV plus IA) were negative for culturable U. parvum. Relative to the results for the control, the levels of expression of interleukin 1β (IL-1β), IL-6, IL-8, and monocyte chemoattractant protein 2 (MCP-2) in fetal skin were significantly decreased in the IV SOLI-only group, the MCP-1 protein concentration in the amniotic fluid was significantly increased in the IV+IA SOLI group, and there was no significant difference in the histological inflammation scoring of lung or chorioamnion among the five groups. In the present study, treatment with either AZ or SOLI (IV only or IV+IA) effectively eradicated macrolide-sensitive U. parvum from the AF. There was no discernible difference in antibiotic therapy efficacy between IV-only and IV

  9. OBSTETRIC AND PERINATAL OUTCOME OF LOW-RISK PREGNANCIES WITH TERM LABOUR AND MECONIUM-STAINED AMNIOTIC FLUID AT ST. PHILOMENA’S HOSPITAL

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    Roopashree D

    2016-12-01

    Full Text Available BACKGROUND Meconium-stained amniotic fluid has been considered as sign of foetal distress in presentations other than breech and is associated with poor foetal outcome, but others consider meconium passage by foetus as physiological phenomenon and procedures of environmental hazards to foetus before birth. MATERIALS AND METHODS 200 women with meconium-stained amniotic fluid in labour, all low-risk pregnancies were included in our study from September 2010 to August 2012 admitted in our department. For uniformity of results, cases with obstetric and medical complications were excluded. The cases were divided into three groups depending upon grades of meconium staining as thin, moderate and thick. Foetal monitoring, uterine contractions and Apgar score, birth weight, resuscitation of baby noted. All babies were followed up to first week of neonatal life. RESULTS Of 200 cases, 147 had caesarean delivery 73.5% and 53 had vaginal delivery 26.5%. The incidence of LSCS was found to be maximum in the thick MSAF group 54.5%, 14% in moderate and 5% in this MSAF group. In moderate MSAF group, 2.17% had <7, 1 minute Apgar. In thick MSAF group, 3.33% had <7 Apgar; in thin MSAF group, 11.76% had <7 Apgar at 1 minute. The p value was 0.094 in all grades of MSAF, the 5 min. Apgar’s were more than 7. The occurrence of complications in the baby did not depend on the mode of delivery. The p value was 0.58, which was insignificant. The ‘p value of association of CTG abnormality and foetal morbidity was significant <0.001. The association of grades of meconium and foetal morbidity, the p value was 0.337, which was insignificant. There was no perinatal mortality in our study. CONCLUSION The incidence of low-risk pregnancies with MSAF in our study was 4.5%. There was increased tendency for LSCS in the MSAF. The foetal morbidity depends on the reactivity to CTG not on grades of MSAF. Therefore, close intrapartum monitoring has to be done in such cases and LSCS done

  10. Lower concentrations of B-vitamin subgroups in the serum and amniotic fluid correlate to cleft lip and palate appearance in the offspring of A/WySn mice.

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    Scheller, Konstanze; Röckl, Thomas; Scheller, Christian; Schubert, Johannes

    2013-09-01

    The pathogenesis and prevention of cleft lip and palate (CL/P) have been studied mainly in clinical and animal experiments. A prophylactic poly-B-vitamin substitution during the first months of pregnancy has provided the most encouraging results for the prevention of CL/P recurrence in families at risk. In vitro studies of the palatal organ in an A/WySn mouse model have confirmed the positive influence of B-vitamins on palatal development. The present animal study was performed to analyze different B-vitamin concentrations in the serum and amniotic fluid of A/WySn mice according to the appearance of CL/P in their offspring. Concentrations of different B-vitamins (B1, B2, B3, B5, B6, and folic acid) in serum and amniotic fluid were analyzed by high-performance liquid chromatographic detection. Immunohistochemical staining against thiamin-1 receptor was performed on histologic midface sections of A/WySn fetuses with (n = 12) and without (n = 14) CL/P. Vitamin B5 (P B5 (P = .4) showed no difference between the 2 groups. Dams with CL/P had significantly lower thiamine concentrations in serum (P = .01) and amniotic fluid (P vitamin subgroups (B1, B5, and folic acid) in amniotic fluid and serum (vitamin B1) was correlated to an increased cleft appearance in A/WySn mice. The high thiamin-1 receptor expression in the palatal tissue of mouse fetuses with CL/P may be caused by a decreased availability of vitamin B1. Copyright © 2013 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.

  11. Electrospun Poly(l-lactide)/Poly(ethylene glycol) Scaffolds Seeded with Human Amniotic Mesenchymal Stem Cells for Urethral Epithelium Repair

    Science.gov (United States)

    Lv, Xiaokui; Guo, Qianping; Han, Fengxuan; Chen, Chunyang; Ling, Christopher; Chen, Weiguo; Li, Bin

    2016-01-01

    Tissue engineering-based urethral replacement holds potential for repairing large segmental urethral defects, which remains a great challenge at present. This study aims to explore the potential of combining biodegradable poly(l-lactide) (PLLA)/poly(ethylene glycol) (PEG) scaffolds and human amniotic mesenchymal cells (hAMSCs) for repairing urethral defects. PLLA/PEG fibrous scaffolds with various PEG fractions were fabricated via electrospinning. The scaffolds were then seeded with hAMSCs prior to implantation in New Zealand male rabbits that had 2.0 cm-long defects in the urethras. The rabbits were randomly divided into three groups. In group A, hAMSCs were grown on PLLA/PEG scaffolds for two days and then implanted to the urethral defects. In group B, only the PLLA/PEG scaffolds were used to rebuild the rabbit urethral defect. In group C, the urethral defect was reconstructed using a regular urethral reparation technique. The repair efficacy was compared among the three groups by examining the urethral morphology, tissue reconstruction, luminal patency, and complication incidence (including calculus formation, urinary fistula, and urethral stricture) using histological evaluation and urethral radiography methods. Findings from this study indicate that hAMSCs-loaded PLLA/PEG scaffolds resulted in the best urethral defect repair in rabbits, which predicts the promising application of a tissue engineering approach for urethral repair. PMID:27517902

  12. Electrospun Poly(l-lactide/Poly(ethylene glycol Scaffolds Seeded with Human Amniotic Mesenchymal Stem Cells for Urethral Epithelium Repair

    Directory of Open Access Journals (Sweden)

    Xiaokui Lv

    2016-08-01

    Full Text Available Tissue engineering-based urethral replacement holds potential for repairing large segmental urethral defects, which remains a great challenge at present. This study aims to explore the potential of combining biodegradable poly(l-lactide (PLLA/poly(ethylene glycol (PEG scaffolds and human amniotic mesenchymal cells (hAMSCs for repairing urethral defects. PLLA/PEG fibrous scaffolds with various PEG fractions were fabricated via electrospinning. The scaffolds were then seeded with hAMSCs prior to implantation in New Zealand male rabbits that had 2.0 cm-long defects in the urethras. The rabbits were randomly divided into three groups. In group A, hAMSCs were grown on PLLA/PEG scaffolds for two days and then implanted to the urethral defects. In group B, only the PLLA/PEG scaffolds were used to rebuild the rabbit urethral defect. In group C, the urethral defect was reconstructed using a regular urethral reparation technique. The repair efficacy was compared among the three groups by examining the urethral morphology, tissue reconstruction, luminal patency, and complication incidence (including calculus formation, urinary fistula, and urethral stricture using histological evaluation and urethral radiography methods. Findings from this study indicate that hAMSCs-loaded PLLA/PEG scaffolds resulted in the best urethral defect repair in rabbits, which predicts the promising application of a tissue engineering approach for urethral repair.

  13. Líquido amniótico tópico: uma potencial nova alternativa para doenças da superfície ocular Topical amniotic fluid: a potential new alternative for ocular surface diseases

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    Guilherme Goulart Quinto

    2008-12-01

    Full Text Available O líquido amniótico banha o feto durante a vida intra-uterina e está em contato permanente com a superfície ocular durante este importante período do desenvolvimento. Ele contém uma série de fatores de crescimento que podem ter diversos efeitos sobre o processo cicatricial. Estes fatores aceleram a recuperação da sensibilidade corneana e regeneração nervosa após procedimentos cerato-refrativos, além de controlar a formação de cicatriz e o equilíbrio da superfície ocular após sua aplicação tópica. Centenas de diferentes proteínas têm sido identificadas no líquido amniótico humano e o papel de cada uma continua desconhecido. Os resultados obtidos até o momento sobre a aplicação de líquido amniótico em doenças de superfície ocular sugerem uma terapia promissora. Pesquisas estão sendo realizadas para identificar os efeitos dos fatores específicos do líquido amniótico sobre a inflamação ocular. O objetivo desta revisão é relatar as propriedades e utilizações atuais do líquido amniótico, bem como apresentar os recentes estudos relacionados ao uso deste líquido e doenças da superfície ocular.Amniotic fluid bathes the fetus during intrauterine life and is in permanent contact with the fetal ocular surface in this important period of development. It contains a series of growth factors that may have multiple effects on the wound healing process. These factors are thought to accelerate the recovery of corneal sensitivity and nerve regeneration after keratorefractive procedures, and also may control scar formation and balance the ocular surface after topical application. Hundreds of different proteins have been identified in the human amniotic fluid, and the role of each still not quite understood. The outcomes obtained so far with amniotic fluid application to ocular surface diseases suggest a promising therapy. Research is underway to identify the effects of specific factors of the amniotic fluid in ocular

  14. Hair Follicle Morphogenesis in the Treatment of Mouse Full-Thickness Skin Defects Using Composite Human Acellular Amniotic Membrane and Adipose Derived Mesenchymal Stem Cells

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    Wu Minjuan

    2016-01-01

    Full Text Available Early repair of skin injury and maximal restoration of the function and appearance have become important targets of clinical treatment. In the present study, we observed the healing process of skin defects in nude mice and structural characteristics of the new skin after transplantation of isolated and cultured adipose derived mesenchymal stem cells (ADMSCs onto the human acellular amniotic membrane (AAM. The result showed that ADMSCs were closely attached to the surface of AAM and grew well 24 h after seeding. Comparison of the wound healing rate at days 7, 14, and 28 after transplantation showed that ADMSCs seeded on AAM facilitated the healing of full-thickness skin wounds more effectively as compared with either hAM or AAM alone, indicating that ADMSCs participated in skin regeneration. More importantly, we noticed a phenomenon of hair follicle development during the process of skin repair. Composite ADMSCs and AAM not only promoted the healing of the mouse full-thickness defects but also facilitated generation of the appendages of the affected skin, thus promoting restoration of the skin function. Our results provide a new possible therapy idea for the treatment of skin wounds with respect to both anatomical regeneration and functional restoration.

  15. Gap Junctions Are Involved in the Rescue of CFTR-Dependent Chloride Efflux by Amniotic Mesenchymal Stem Cells in Coculture with Cystic Fibrosis CFBE41o- Cells

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    Annalucia Carbone

    2018-01-01

    Full Text Available We previously found that human amniotic mesenchymal stem cells (hAMSCs in coculture with CF immortalised airway epithelial cells (CFBE41o- line, CFBE on Transwell® filters acquired an epithelial phenotype and led to the expression of a mature and functional CFTR protein. In order to explore the role of gap junction- (GJ- mediated intercellular communication (GJIC in this rescue, cocultures (hAMSC : CFBE, 1 : 5 ratio were studied for the formation of GJIC, before and after silencing connexin 43 (Cx43, a major component of GJs. Functional GJs in cocultures were inhibited when the expression of the Cx43 protein was downregulated. Transfection of cocultures with siRNA against Cx43 resulted in the absence of specific CFTR signal on the apical membrane and reduction in the mature form of CFTR (band C, and in parallel, the CFTR-dependent chloride channel activity was significantly decreased. Cx43 downregulation determined also a decrease in transepithelial resistance and an increase in paracellular permeability as compared with control cocultures, implying that GJIC may regulate CFTR expression and function that in turn modulate airway epithelium tightness. These results indicate that GJIC is involved in the correction of CFTR chloride channel activity upon the acquisition of an epithelial phenotype by hAMSCs in coculture with CF cells.

  16. Second trimester amniotic fluid glucose, uric acid, phosphate, potassium, and sodium concentrations in relation to maternal pre-pregnancy BMI and birth weight centiles.

    Science.gov (United States)

    Fotiou, Maria; Michaelidou, Alexandra Maria; Athanasiadis, Apostolos P; Menexes, Georgios; Symeonidou, Maria; Koulourida, Vasiliki; Ganidou, Maria; Theodoridis, Theodoros D; Tarlatzis, Basil C

    2015-05-01

    To study the evolution profile of amniotic fluid (AF) glucose, uric acid, phosphate, potassium, and sodium, in the second trimester of pregnancy, and explore the possible relations between the concentration of these components and maternal, as well as neonatal characteristics. AF of 52 pregnant women was analyzed using an automatic multichannel analyzer. Maternal age, pre-pregnancy Body Mass Index (BMI), inter-pregnancy intervals, and smoking status were derived from questionnaires. Information on pregnancy and delivery was collected from medical records. Uric acid increased (r = 0.423, p uric acid concentration (r = 0.460, p uric acid and phosphate levels were significantly related to birth weight centiles (R(2)( )= 0.345, p uric acid concentration, and (c) in appropriate for gestational age infants, AF phosphate and uric acid levels may serve as potential biomarkers of birth weight centiles. Further studies on AF composition may help to unravel the biochemical pathways underlying fetal development and could offer insight on the potential impact of maternal nutritional management on fetal growth regulation.

  17. Women's expectations and experiences of rupture of membranes and views of the potential use of reagent pads for detecting amniotic fluid.

    Science.gov (United States)

    Spiby, Helen; Borrelli, Sara; Hughes, Anita J

    2017-12-01

    To explore first-time mothers' expectations and experiences regarding rupture of membranes at term and their views on the potential use of reagent pads that detect amniotic fluid. There is little information available on women's experiences of spontaneous rupture of membranes, or interest in using methods to confirm rupture of membranes (e.g. reagent pads). Descriptive qualitative study, using focus groups and telephone interviews with women during pregnancy and after the birth of their first baby. Thematic analysis was undertaken to analyse women's responses. Ethics committee approval was obtained. Twenty-five women participated in the study of whom 13 contributed both during pregnancy and postpartum between October 2015-March 2016. Three overarching themes were identified from the data from women's expectations and experiences: uncertainty in how, when and where membranes may rupture; information which was felt to be limited and confirmation of rupture of membranes. The potential use of reagent pads met with varied responses. Women were interested in having facts and figures regarding rupture of membranes, such as characteristics of liquor; volume and probability of membranes rupturing spontaneously at term. Use of a pad as a means of confirmation was viewed as helpful, although the potential for increasing anxiety was raised. © 2017 John Wiley & Sons Ltd.

  18. Reference intervals for N-terminal pro-B-type natriuretic peptide in amniotic fluid between 10 and 34 weeks of gestation.

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    Waltraut M Merz

    Full Text Available BACKGROUND: In adult and pediatric cardiology, n-terminal pro-B-type natriuretic peptide (nt-proBNP serves as biomarker in the diagnosis and management of cardiovascular dysfunction. Elevated levels of circulating nt-proBNP are present in fetal conditions associated with myocardial pressure or volume load. Compared to fetal blood sampling, amniocentesis is technically easier and can be performed from early pregnancy onwards. We aimed to investigate amniotic fluid (AF nt-proBNP concentrations in normal pregnancies between 10 and 34 weeks of gestation. METHODS: Nt-proBNP and total protein (TP was measured in AF by chemiluminescence assay (photometry, respectively. To adjust for a potential dilutional effect, the AF-nt-proBNP/AF-TP ratio was analyzed. Reference intervals were constructed by regression modeling across gestational age. RESULTS: 132 samples were analyzed. A negative correlation between AF-nt-proBNP/AF-TP ratio and gestational age was observed. Curves for the mean and the 5% and 95% reference interval between 10 and 34 weeks of gestation were established. CONCLUSION: In normal pregnancy, nt-proBNP is present in AF and decreases during gestation. Our data provide the basis for research on AF-nt-proBNP as biomarker in fetal medicine.

  19. Amniotic Fluid Embolism Pathophysiology Suggests the New Diagnostic Armamentarium: β-Tryptase and Complement Fractions C3-C4 Are the Indispensable Working Tools

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    Francesco Paolo Busardò

    2015-03-01

    Full Text Available Amniotic fluid embolism (AFE is an uncommon obstetric condition involving pregnant women during labor or in the initial stages after delivery. Its incidence is estimated to be around 5.5 cases per 100,000 deliveries. Therefore, this paper investigated the pathophysiological mechanism, which underlies AFE, in order to evaluate the role of immune response in the development of this still enigmatic clinical entity. The following databases (from 1956 to September 2014 Medline, Cochrane Central, Scopus, Web of Science and Science Direct were used, searching the following key words: AFE, pathophysiology, immune/inflammatory response, complement and anaphylaxis. The main key word “AFE” was searched singularly and associated individually to each of the other keywords. Of the 146 sources found, only 19 were considered appropriate for the purpose of this paper. The clinical course is characterized by a rapid onset of symptoms, which include: acute hypotension and/or cardiac arrest, acute hypoxia (with dyspnoea, cyanosis and/or respiratory arrest, coagulopathies (disseminated intravascular coagulation and/or severe hemorrhage, coma and seizures. The pathology still determines a significant morbidity and mortality and potential permanent neurological sequelae for surviving patients. At this moment, numerous aspects involving the pathophysiology and clinical development are still not understood and several hypotheses have been formulated, in particular the possible role of anaphylaxis and complement. Moreover, the detection of serum tryptase and complement components and the evaluation of fetal antigens can explain several aspects of immune response.

  20. Lower concentrations of receptor for advanced glycation end products and epiregulin in amniotic fluid correlate to chemically induced cleft palate in mice.

    Science.gov (United States)

    Wang, Xinhuan; Zhu, Jingjing; Fang, Yanjun; Bian, Zhuan; Meng, Liuyan

    2017-04-01

    This study investigated the correlation between differentially expressed proteins in amniotic fluid (AF) and cleft palate induced by all-trans retinoic acid (atRA), and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in mice. Seven proteins were differentially expressed at embryonic day (E) 16.5 in atRA and control groups as revealed by label-based mouse antibody array. Enzyme-linked immunosorbent assay was further used to detect the expression levels of these proteins in AF from E13.5 to E16.5 in atRA, TCDD, and control groups. The cleft palate groups showed lower concentrations of receptor for advanced glycation end products (RAGE) and epiregulin at E16.5. RAGE immunostaining obviously decreased in palatal tissue sections obtained from E14.5 to E16.5 in the cleft palate groups as revealed by immunohistochemistry. These findings indicate that reduced levels of RAGE and epiregulin in AF are correlated to chemically induced cleft palate in mice. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Placental growth hormone, pituitary growth hormone, insulin-like growth factor, and ghrelin in umbilical cord blood serum and amniotic fluid.

    Science.gov (United States)

    Kędzia, Andrzej; Petriczko, Elżbieta; Tarka, Agata

    2013-01-01

    In the search for biomarkers that allow the prediction of neonatal growth and development, placental growth hormone(PGH), pituitary growth hormone (GH1), insulin-like growth factor 1 (IGF-1), and ghrelin concentrations were assessed in the amniotic fluid and in the umbilical cord blood of 92 neonates. The proteins were assayed by the ELISA method. Their concentration values were compared in 57 full-term neonates and 35 prematurely born neonates, as well as in both large (> 4,000 g) and small neonates (hormones studied did not show statistically significant differences. A distinct tendency was noticed towards an increase in PGH concentration in both prematurely born and small neonates. In large neonates, statistically significantly higher IGF-1 concentrations were found compared to the prematurely born neonates. Our studies indicate an important role for PGH in maintaining a proper IGF-1 pool and demonstrate the existence of a direct influence on the function of the placenta in prematurely born neonates through the activation of compensation mechanisms,which stimulate IGF-1 synthesis.

  2. Higher levels of ethyl paraben and butyl paraben in rat amniotic fluid than in maternal plasma after subcutaneous administration

    DEFF Research Database (Denmark)

    Frederiksen, Hanne; Taxvig, Camilla; Hass, Ulla

    2008-01-01

    were analyzed by liquid chromatography-tandem mass spectrometry. Markedly higher levels of ethyl paraben compared to butyl paraben were found in all fluids and tissues. Both ethyl paraben and butyl paraben in maternal plasma, livers, and whole-body tissues from fetus seemed to be saturated after dosing...

  3. Psychobiology of the amniotic environment.

    Science.gov (United States)

    Benassi, Luigi; Accorsi, Francesca; Marconi, Lorenza; Benassi, Gianluca

    2004-01-01

    Water, basic element of amniotic fluid (A.F.), is closely related to Life, Fertility and Motherhood in several cultures and religions. Through material evidences of an essential growth medium and useful diagnostic source, a new concept grow up: the fluid as a first real environment in which fetus lives and acts. Many studies confirm that in A.F. fetus starts his character-building, his memory and his intelligence. The fluid seems to be the first means of learning and acknowledgement. Sounds, smells and tastes are perceived as well as emotions and fears. Urinoterapy and staminal cells sampling shows how A.F. can be considered as an additional terapeutic resource.

  4. Infecção Assintomática do Líquido Amniótico Asymptomatic Amniotic Fluid Infection

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    Roxeane Martins Monteiro

    2002-01-01

    Full Text Available Objetivo: determinar a presença de infecção assintomática do líquido amniótico em gestantes, identificar os agentes bacterianos envolvidos na infecção e determinar o perfil de suscetibilidade a antimicrobianos in vitro. Métodos: foram obtidas 81 amostras de líquido amniótico, colhidas por amniocentese, em gestantes sem sinais de trabalho de parto e sem suspeita clínica de infecção, atendidas na Maternidade Escola Assis Chateaubriand, entre agosto/97 e janeiro/99. Pesquisou-se a presença de bactérias aeróbias, anaeróbias estritas/facultativas e micoplasmas genitais. As bactérias anaeróbias foram identificadas pelo sistema ATBÒ (Bio-Mérieux e os micoplasmas pelo kit Micoplasmas ISTÒ (Bio-Mérieux. Resultados: entre as amostras obtidas, oito (9,8% apresentaram culturas positivas, sendo que em duas foram identificadas duas espécies bacterianas. Os patógenos isolados foram: Ureaplasma urealyticum (7 casos, 8,6%, Mycoplasma hominis (1 caso, 1,2% e Peptostreptococcus sp (2 casos 2,4%. O padrão de resistência aos antimicrobianos caracterizou-se pela maior resistência dos micoplasmas à eritromicina (37,5% e nenhuma resistência às ciclinas. Conclusões: o percentual de infecções assintomáticas foi muito elevado, havendo necessidade de serem realizadas novas pesquisas para avaliar as conseqüências da infecção subclínica nas grávidas e em seus conceptos, que envolvam métodos que identifiquem micoplasmas genitais, já que foram as bactérias mais freqüentemente isoladas.Purpose: to determine the presence of asymptomatic amniotic fluid infection in pregnant women, to identify the bacterial agents involved in the infection and to determine the antimicrobial susceptibility in vitro. Methods: amniotic fluid samples were obtained by amniocentesis from 81 pregnant women without labor signs and without suspucion of clinical infection, attended at Maternidade Escola Assis Chateaubriand from August 1997 to January 1999. The

  5. Amnioinfusão em trabalho de parto com líquido meconiado Amnioinfusion during labor with meconium-stained amniotic fluid

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    Ricardo F. Savaris

    1999-08-01

    Full Text Available Objetivo: relatar a experiência com o uso da técnica de amnioinfusão nas pacientes em trabalho de parto que apresentam mecônio e verificar a incidência de complicações, da síndrome da aspiração do mecônio com o uso desse procedimento e de cesáreas. Método: foram estudadas retrospectivamente 20 gestantes a termo em trabalho de parto, com líquido amniótico meconiado no Centro Obstétrico de duas instituições do Sistema Único de Saúde no Estado do Rio Grande do Sul. A infusão inicial de 1000 ml de soro fisiológico não-aquecido foi feita a velocidade de 20 a 30 ml por minuto, depois reduzida para 3 ml por minuto. O líquido era drenado com a elevação da apresentação cefálica. Resultados: a amnioinfusão demonstrou-se de fácil aplicação, quando foi utilizado um cateter nasogástrico. Nenhuma paciente apresentou complicações importantes relacionadas com o procedimento. Não houve presença de mecônio abaixo das cordas vocais nos recém-nascidos. A incidência de parto cesárea foi de 3/20 (15%. Conclusões: a amnioinfusão é uma técnica de baixo custo e factível que não apresentou complicações nos casos relatados.Purpose: to report the personal experience with the use of the amnioinfusion technique in patients in labor presenting meconial amniotic fluid, and the incidence of complications, the meconium aspiration syndrome and of cesarean sections. Method: twenty patients at term and in labor with meconial amniotic fluid were evaluated retrospectively, at the delivery ward at two public institutions of Rio Grande do Sul. An initial infusion of 1.000 ml of normal saline solution at room temperature, at an infusion rate of 20-30 ml per minute was initiated and then reduced to 3 ml per minute. The liquid was drained by elevating the cephalic pole. Results: the procedure was feasible when a nasogastric catheter was used. None of the patients presented major complications related to the procedure. None of the neonates

  6. Significant performance variation among PCR systems in diagnosing congenital toxoplasmosis in São Paulo, Brazil: analysis of 467 amniotic fluid samples

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    Thelma Suely Okay

    2009-03-01

    Full Text Available INTRODUCTION: Performance variation among PCR systems in detecting Toxoplasma gondii has been extensively reported and associated with target genes, primer composition, amplification parameters, treatment during pregnancy, host genetic susceptibility and genotypes of different parasites according to geographical characteristics. PATIENTS: A total of 467 amniotic fluid samples from T. gondii IgM- and IgG-positive Brazilian pregnant women being treated for 1 to 6 weeks at the time of amniocentesis (gestational ages of 14 to 25 weeks. METHODS: One nested-B1-PCR and three one-round amplification systems targeted to rDNA, AF146527 and the B1 gene were employed. RESULTS: Of the 467 samples, 189 (40.47% were positive for one-round amplifications: 120 (63.49% for the B1 gene, 24 (12.69% for AF146527, 45 (23.80% for both AF146527 and the B1 gene, and none for rDNA. Fifty previously negative one-round PCR samples were chosen by computer-assisted randomization analysis and re-tested (nested-B1-PCR, during which nine additional cases were detected (9/50 or 18%. DISCUSSION: The B1 gene PCR was far more sensitive than the AF146527 PCR, and the rDNA PCR was the least effective even though the rDNA had the most repetitive sequence. Considering that the four amplification systems were equally affected by treatment, that the amplification conditions were optimized for the target genes and that most of the primers have already been reported, it is plausible that the striking differences found among PCR performances could be associated with genetic diversity in patients and/or with different Toxoplasma gondii genotypes occurring in Brazil. CONCLUSION: The use of PCR for the diagnosis of fetal Toxoplasma infections in Brazil should be targeted to the B1 gene when only one gene can be amplified, preferably by nested amplification with primers B22/B23.

  7. Elevated second-trimester maternal serum β-human chorionic gonadotropin and amniotic fluid alpha-fetoprotein as indicators of adverse obstetric outcomes in fetal Turner syndrome.

    Science.gov (United States)

    Alvarez-Nava, Francisco; Soto, Marisol; Lanes, Roberto; Pons, Hector; Morales-Machin, Alisandra; Bracho, Ana

    2015-12-01

    The objective of this study was to determine the ability of biochemical analytes to identify adverse outcomes in pregnancies with Turner syndrome. Maternal serum and amniotic fluid (AF) marker concentrations were measured in 73 singleton pregnancies with Turner syndrome (10-22 weeks of gestation). Fetal Turner syndrome was definitively established by cytogenetic analysis. Two subgroups, fetuses with hydrops fetalis versus fetuses with cystic hygroma, were compared. Receiver operating characteristic curves and relative risk were established for a cut-off multiples of the median ≥3.5 for β-subunit of human chorionic gonadotropin (hCG) or AF alpha-fetoprotein (AFP). Forty-nine (67%) of 73 pregnant women had an abnormal maternal serum. While levels of pregnancy-associated plasma protein-A and free β-subunit (fβ)-hCG were not different to those of the control group, AFP, unconjugated estriol and β-hCG concentrations were significantly different in the study group (P < 0.05), when compared to those of unaffected pregnancies. Levels of β-hCG in pregnancies with hydrops fetalis were significantly higher than in those with cystic hygroma (P <0.0001), as were AF-AFP concentrations (P <0.0015). In addition, abnormalities in both maternal serum β-hCG and AF-AFP predicted fetal death. The relative risk of adverse obstetric outcome was 10.667 (P = 0.0004; 95% confidence interval [CI]: 1.554-73.203) for β-hCG and 2.19 (P = 0.0256; 95% CI: 1.001 to 4.779), for AF-AFP. Maternal serum β-hCG and AF-AFP levels may preferentially identify those Turner syndrome pregnancies with the highest risk of fetal death. © 2015 Japan Society of Obstetrics and Gynecology.

  8. Respiratory distress including meconium aspiration syndrome in vigorous neonates born through meconium stained amniotic fluid: incidence, onset, severity and predictors at birth.

    Science.gov (United States)

    Singh, S N; Srivastava, Roli; Singh, Anita; Tahazzul, M; Kumar, Mala; Kanta, Chandra; Chandra, S

    2013-07-01

    This study aimed to find out incidence, predictors, onset and severity of respiratory distress including meconium aspiration syndrome (MAS) among vigorous neonates born through meconium stained amniotic fluid (MSAF), which may or may not be evident at birth. Two hundred ninety vigorous neonates were studied. Data were collected on perinatal risk factors, clinical course and development of respiratory distress. Predictors of respiratory distress were identified by logistic regression and a score based on adjusted OR was assigned for each. Diagnostic performance of the score (0-24) was assessed on another 247 vigorous neonates using receiver operator characteristic analysis (ROC). Respiratory distress developed in 97(33.4 %) infants, MAS in 75(25.9 %). The distress appeared within 12 h in 97.9 %, was severe in only 21.7 %. Of 10 risk factors significantly associated with respiratory distress, seven entered in regression analysis. Fetal distress(adj OR = 11.8; 95%CI = 6.2-22.5), prolonged labor(adj OR = 5.2; 95%CI = 2.5-10.7), and absent/poor cry(adj OR = 5.6; 95%CI = 2.4-13.3) were identified as independent predictors; each assigned a score of 12, 6 and 6, respectively. To predict respiratory distress, a cut-off score of 9 points had sensitivity-74.1 % (95%CI = 63.3 %-82.7 %), specificity-84.6 % (95 % CI = 77.9 %-89.6 %), positive predictive value- 71.6 % (95%CI = 60.8 %-80.4 %), negative predictive value- 86.2 % (95 % CI = 79.6 %-90.9 %), likelihood ratio (LR) + ve 4.8(95%CI = 3.3-7.0) and LR-ve 0.3(95%CI = 0.2-0.4). Respiratory distress occurred in one third neonates, mostly had onset within 12 h of birth, and it was mild to moderate in majority. Fetal distress, prolonged labor, and absent/poor cry predicted respiratory distress and were validated. However, larger studies in different settings are required to confirm its utility.

  9. [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.

  10. 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.

  11. Liquid chromatography--tandem mass spectrometry analysis of cocaine and its metabolites from blood, amniotic fluid, placental and fetal tissues: study of the metabolism and distribution of cocaine in pregnant rats.

    Science.gov (United States)

    Srinivasan, K; Wang, P P; Eley, A T; White, C A; Bartlett, M G

    2000-08-18

    The ability to simultaneously quantitate cocaine and its 12 metabolites from pregnant rat blood, amniotic fluid, placental and fetal tissue homogenates aids in elucidating the metabolism and distribution of cocaine. An efficient extraction method was developed to simultaneously recover these 13 components using underivatized silica solid-phase extraction (SPE) cartridges. The overall recoveries for cocaine and its metabolites were studied from pregnant rat blood (47-100%), amniotic fluid (61-100%), placental homogenate (31-83%), and fetal homogenate (39-87%). Extraction of the samples using silica is not classical SPE, but rather allows for the concentration of the sample into a small volume prior to injection and the removal of the proteins due to their strong interaction with the active silica surface. A positive ion mode electrospray ionization liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was used and validated to simultaneously quantitate cocaine and 12 metabolites from these four biological matrices. A gradient elution method with a Zorbax XDB C8 reversed-phase column was used to separate the components. Multiple reaction monitoring (MRM) of a product ion arising from the corresponding precursor ion was used in order to enhance the selectivity and sensitivity of the method. Low background noise was observed from the complex biological matrices due to efficient SPE and the selectivity of the MRM mode. Linear calibration curves were generated from 0.01 to 2.50 ppm. The method also showed high intra-day (n =3) and inter-day (n=9) precision (% RSD) and accuracy (% error) for all components. The limits of detection (LODs) for the method ranged from 0.15 to 10 ppb. The LODs of cocaine and its major metabolites were less than 1 ppb from all four biological matrices. This method was applied to the study of the metabolism and distribution of cocaine in pregnant rats following intravenous infusion to a steady state plasma drug concentration. The

  12. Acceleration of Regeneration of Large Gap Peripheral Nerve Injuries Using Acellular Nerve Allografts plus amniotic Fluid Derived Stem Cells (AFS)

    Science.gov (United States)

    2016-09-01

    seeded constructs were used to repair critical-sized, large gap nerve injury in rats and their functional recovery was monitored longitudinally using... injuries in non-human primates. These represent a more translational model of peripheral nerve injury and repair . 15. SUBJECT TERMS 16. SECURITY...AWARD NUMBER: W811XWH-13-1-0310 TITLE: Acceleration of Regeneration of Large-Gap Peripheral Nerve Injuries Using Acellular Nerve Allografts

  13. Serum-Free Cryopreservation of Human Amniotic Epithelial Cells

    OpenAIRE

    H. Niknejad; H. Peirovi; B. Jambar Noushin

    2013-01-01

    Introduction & Objective: One of the important issues in long term storage of cells is removal of animal serum from cell culture environments. The aim of this study was to evaluate amni-otic fluid (AF), which is full of growth factors, as substitute for fetal bovine serum (FBS) in the cryopreservation protocol. Materials & Methods: In this experimental study human amniotic epithelial cells were isolated from placentas which were seronegative for microbial infections. The cells were preserved ...

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

    Directory of Open Access Journals (Sweden)

    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.

  15. miR-145 modulates lncRNA-ROR and Sox2 expression to maintain human amniotic epithelial stem cell pluripotency and β islet-like cell differentiation efficiency.

    Science.gov (United States)

    Zou, Gang; Liu, Te; Guo, Lihe; Huang, Yongyi; Feng, Ya; Huang, Qin; Duan, Tao

    2016-10-10

    In this study, we observed a great reduction in the expression of the endogenous long noncoding RNA ROR (lncRNA-ROR) and the stem cell transcription factor Sox2, in contrast to a marked increase in miR-145 expression, during the course of in vitro induced differentiation of human amniotic epithelial stem cells (HuAECs). Bioinformatics analysis and the luciferase reporter assay revealed binding of miR-145 to specific sites in lncRNA-ROR and Sox2, silencing their expression. Overexpression of a lncRNA-ROR-specific siRNA effectively downregulated the expression levels of Sox2 and other stem cell markers in HuAECs while weakening the efficiency of HuAEC differentiation into β islet-like cells. Moreover, the in vitro response of HuAEC-derived β islet-like cells to extracellular stimuli and C-peptide release by these cells were markedly weakened in the siRNA-ROR transfection group. Furthermore, the in vivo expression of β islet-like cell biomarkers was substantially reduced in HuAECs in the siRNA-ROR transfection group, and their in vivo β islet-like cell differentiation and insulin release capacities were reduced in a streptozocin-induced diabetic rat model. The experimental results indicate that lncRNA-ROR effectively maintains Sox2 gene expression through competitive binding to miR-145, achieving pluripotency maintenance in HuAECs and regulation of their directed β islet-like cell differentiation efficiency. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. A new method for immediate derivatization of hydroxyl groups by fluoroalkyl chloroformates and its application for the determination of sterols and tocopherols in human serum and amniotic fluid by gas chromatography-mass spectrometry.

    Science.gov (United States)

    Řimnáčová, L; Hušek, P; Šimek, P

    2014-04-25

    A new method has been described for efficient derivatization of secondary alicyclic hydroxyl groups in steroids by alkyl chloroformates (RCFs). Cholesterol, an essential human sterol and a steroid precursor in eukaryotic cells, was used as a model for treatment with various RCFs in an aqueous and non-aqueous environment. While the cholesterol hydroxyl group did not react completely with any of the tested RCFs reagents in the former case, trifluoroethyl chloroformate (TFECF) or heptafluorobutyl chloroformate (HFBCF) fully converts cholesterol and related metabolites into the corresponding mixed carbonates under anhydrous conditions in seconds. The acylation reaction was combined with liquid-liquid microextraction (LLME) between isooctane and acetonitrile phase. The sample preparation requires just a stepwise addition of 50μl isooctane with 5μl of a pyridine catalyst, 100μl acetonitrile and 100μl isooctane with dissolved 5μl of the fluoroalkyl chloroformate reagent to a dried sample. The protocol developed in this study was successfully tested for GC-MS analysis of 12 important model steroids and four main tocopherols. Each analyte provided a single peak with excellent GC separation properties, well defined EI spectra containing diagnostic fragment ions suitable for their identification and quantitation. The new method was further validated for the determination of six diagnostic non-cholesterol sterols and four main tocopherols in human serum and in amniotic fluid. Satisfactory data were obtained in terms of calibration, quantitation limits (for sterols and tocopherols, 0.05 and 0.15μg/ml, respectively), within-run precision (0.9-19.5%) and between-run precision (0.2-19.0%), accuracy (82-115%) and recovery (90-110%). The validated method was successfully applied to GC-MS analysis of the analytes in woman sera and amniotic fluids and the results are well-comparable with those reported by other authors. The presented work demonstrates for the first time

  17. Amniotic oxytocin and vasopressin in relation to human fetal development and labour

    NARCIS (Netherlands)

    Oosterbaan, H. P.; Swaab, D. F.

    1989-01-01

    Previous experiments in rats revealed increased amniotic oxytocin (OXT) levels in the course of normal development and increased vasopressin (AVP) levels in retarded fetal growth. In order to see whether similar changes would also occur in human, OXT and AVP levels were determined in amniotic fluid,

  18. Mesenchymal Stem Cells Obtained from Synovial Fluid Mesenchymal Stem Cell-Derived Induced Pluripotent Stem Cells on a Matrigel Coating Exhibited Enhanced Proliferation and Differentiation Potential

    OpenAIRE

    Zheng, Yu-Liang; Sun, Yang-Peng; Zhang, Hong; Liu, Wen-Jing; Jiang, Rui; Li, Wen-Yu; Zheng, You-Hua; Zhang, Zhi-Guang

    2015-01-01

    Induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs) serve as a promising source for cell-based therapies in regenerative medicine. However, optimal methods for transforming iPSCs into MSCs and the characteristics of iPSC-MSCs obtained from different methods remain poorly understood. In this study, we developed a one-step method for obtaining iPSC-MSCs (CD146+STRO-1+ MSCs) from human synovial fluid MSC-derived induced iPSCs (SFMSC-iPSCs). CD146-STRO-1-SFMSCs were reprogram...

  19. A Three-Dimensional Skeletal Reconstruction of the Stem Amniote Orobates pabsti (Diadectidae: Analyses of Body Mass, Centre of Mass Position, and Joint Mobility.

    Directory of Open Access Journals (Sweden)

    John A Nyakatura

    Full Text Available Orobates pabsti, a basal diadectid from the lower Permian, is a key fossil for the understanding of early amniote evolution. Quantitative analysis of anatomical information suffers from fragmentation of fossil bones, plastic deformation due to diagenetic processes and fragile preservation within surrounding rock matrix, preventing further biomechanical investigation. Here we describe the steps taken to digitally reconstruct MNG 10181, the holotype specimen of Orobates pabsti, and subsequently use the digital reconstruction to assess body mass, position of the centre of mass in individual segments as well as the whole animal, and study joint mobility in the shoulder and hip joints. The shape of most fossil bone fragments could be recovered from micro-focus computed tomography scans. This also revealed structures that were hitherto hidden within the rock matrix. However, parts of the axial skeleton had to be modelled using relevant isolated bones from the same locality as templates. Based on the digital fossil, mass of MNG 10181 was estimated using a model of body shape that was varied within a plausible range to account for uncertainties of the dimension. In the mean estimate model the specimen had an estimated mass of circa 4 kg. Varying of the mass distribution amongst body segments further revealed that Orobates carried most of its weight on the hind limbs. Mostly unrestricted joint morphology further suggested that MNG 10181 was able to effectively generate propulsion with the pelvic limbs. The digital reconstruction is made available for future biomechanical studies.

  20. Serum-Free Cryopreservation of Human Amniotic Epithelial Cells

    Directory of Open Access Journals (Sweden)

    H. Niknejad

    2013-04-01

    Full Text Available Introduction & Objective: One of the important issues in long term storage of cells is removal of animal serum from cell culture environments. The aim of this study was to evaluate amni-otic fluid (AF, which is full of growth factors, as substitute for fetal bovine serum (FBS in the cryopreservation protocol. Materials & Methods: In this experimental study human amniotic epithelial cells were isolated from placentas which were seronegative for microbial infections. The cells were preserved in 24 different patterns for 12 months in -196 ?C (liquid nitrogen and viability of cells were determined before and after cryopreservation by trypan blue and MTT assay. Moreover, Oct-4 expression was studied to determine pluripotency before and after cryopreservation with immunocytochemistry. Results were compared between groups with ANOVA (Tukey Post-Test. P.value under 0.01 and 0.05 was considered statistically significant. Results: The presence of DMEM, FBS or AF is necessary for amniotic cell cryopreservation. Trypan-blue, MTT and immunocytochemistry showed that there isn’t significant difference between using AF and FBS in viability and pluripotency of cells. Moreover, results showed that DMSO is a better cryoprotectant compared to glycerol. Conclusion : Results showed that amniotic fluid can be a proper substitute for FBS in amniotic epithelial cells cryopreservation. (Sci J Hamadan Univ Med Sci 2013; 20 (1:15-24

  1. The CT appearances of delayed amniotic fluid clearance from the lungs in an infant with absent pulmonary valve and congenital lobar emphysema

    International Nuclear Information System (INIS)

    Fink, A. Michelle; Edis, Brian; Massie, John

    2005-01-01

    Congenital lobar emphysema (CLE) is a cause of severe neonatal respiratory distress. Overexpansion of the affected pulmonary lobe in the fetus is due to narrowing of the airway, with a resultant 'ball-valve' effect. At birth, there may be delayed clearance of fetal lung fluid. Early chest radiographs show opacification of the hyperexpanded lobe. The CT findings in the immediate neonatal period have not been previously reported. We describe the imaging in a neonate with tetralogy of Fallot and absent pulmonary valve with secondary CLE. CT demonstrates the hyperexpanded lobe with initial thickening of the interlobular septa and alveolar ground glass attenuation, with subsequent clearing. This resorption of fetal lung fluid via the pulmonary interstitium should not be confused with interstitial lung disease. (orig.)

  2. The CT appearances of delayed amniotic fluid clearance from the lungs in an infant with absent pulmonary valve and congenital lobar emphysema

    Energy Technology Data Exchange (ETDEWEB)

    Fink, A. Michelle [Royal Children' s Hospital, Department of Medical Imaging, Parkville, Victoria (Australia); University of Melbourne, Melbourne, Victoria (Australia); Edis, Brian [Royal Children' s Hospital, Department of Cardiology, Parkville, Victoria (Australia); Massie, John [University of Melbourne, Melbourne, Victoria (Australia); Royal Children' s Hospital, Department of Respiratory Medicine, Parkville, Victoria (Australia); Murdoch Children' s Research Institute, Melbourne, Victoria (Australia)

    2005-09-01

    Congenital lobar emphysema (CLE) is a cause of severe neonatal respiratory distress. Overexpansion of the affected pulmonary lobe in the fetus is due to narrowing of the airway, with a resultant 'ball-valve' effect. At birth, there may be delayed clearance of fetal lung fluid. Early chest radiographs show opacification of the hyperexpanded lobe. The CT findings in the immediate neonatal period have not been previously reported. We describe the imaging in a neonate with tetralogy of Fallot and absent pulmonary valve with secondary CLE. CT demonstrates the hyperexpanded lobe with initial thickening of the interlobular septa and alveolar ground glass attenuation, with subsequent clearing. This resorption of fetal lung fluid via the pulmonary interstitium should not be confused with interstitial lung disease. (orig.)

  3. MRI of the brain stem using fluid attenuated inversion recivery pulse sequences

    International Nuclear Information System (INIS)

    De Coene, B.; Hajnal, J.V.; Pennock, J.M.; Bydder, G.M.

    1993-01-01

    Heavily T2-weighted fluid-attenuated inversion recovery (FLAIR) sequences with inversion times of 2000-2500 ms and echo times of 130-200 ms were used to image the brain stem of a normal adult and five patients. These sequences produce high signal from many white matter tracts and display high lesion contrast. The corticospinal and parietopontine tracts, lateral and medial lemnisci, superior and inferior cerebellar peduncles, medial longitudinal fasciculi, thalamo-olivary tracts the cuneate and gracile fasiculi gave high signal and were directly visualised. The oculomotor and trigeminal nerves were demonstrated within the brain stem. Lesions not seen with conventional T2-weighted spin echo sequences were seen with high contrast in patients with infarction, multiple sclerosis, sarcoidosis, chunt obstruction and metastatic tumour. The anatomical detail and high lesion contrast given by the FLAIR pulse sequence appear likely to be of value in diagnosis of disease in the brain stem. (orig.)

  4. A Simple Alkaline Method for Decellularizing Human Amniotic Membrane for Cell Culture

    Science.gov (United States)

    Saghizadeh, Mehrnoosh; Winkler, Michael A.; Kramerov, Andrei A.; Hemmati, David M.; Ghiam, Chantelle A.; Dimitrijevich, Slobodan D.; Sareen, Dhruv; Ornelas, Loren; Ghiasi, Homayon; Brunken, William J.; Maguen, Ezra; Rabinowitz, Yaron S.; Svendsen, Clive N.; Jirsova, Katerina; Ljubimov, Alexander V.

    2013-01-01

    Human amniotic membrane is a standard substratum used to culture limbal epithelial stem cells for transplantation to patients with limbal stem cell deficiency. Various methods were developed to decellularize amniotic membrane, because denuded membrane is poorly immunogenic and better supports repopulation by dissociated limbal epithelial cells. Amniotic membrane denuding usually involves treatment with EDTA and/or proteolytic enzymes; in many cases additional mechanical scraping is required. Although ensuring limbal cell proliferation, these methods are not standardized, require relatively long treatment times and can result in membrane damage. We propose to use 0.5 M NaOH to reliably remove amniotic cells from the membrane. This method was used before to lyse cells for DNA isolation and radioactivity counting. Gently rubbing a cotton swab soaked in NaOH over the epithelial side of amniotic membrane leads to nearly complete and easy removal of adherent cells in less than a minute. The denuded membrane is subsequently washed in a neutral buffer. Cell removal was more thorough and uniform than with EDTA, or EDTA plus mechanical scraping with an electric toothbrush, or n-heptanol plus EDTA treatment. NaOH-denuded amniotic membrane did not show any perforations compared with mechanical or thermolysin denuding, and showed excellent preservation of immunoreactivity for major basement membrane components including laminin α2, γ1-γ3 chains, α1/α2 and α6 type IV collagen chains, fibronectin, nidogen-2, and perlecan. Sodium hydroxide treatment was efficient with fresh or cryopreserved (10% dimethyl sulfoxide or 50% glycerol) amniotic membrane. The latter method is a common way of membrane storage for subsequent grafting in the European Union. NaOH-denuded amniotic membrane supported growth of human limbal epithelial cells, immortalized corneal epithelial cells, and induced pluripotent stem cells. This simple, fast and reliable method can be used to standardize

  5. A simple alkaline method for decellularizing human amniotic membrane for cell culture.

    Directory of Open Access Journals (Sweden)

    Mehrnoosh Saghizadeh

    Full Text Available Human amniotic membrane is a standard substratum used to culture limbal epithelial stem cells for transplantation to patients with limbal stem cell deficiency. Various methods were developed to decellularize amniotic membrane, because denuded membrane is poorly immunogenic and better supports repopulation by dissociated limbal epithelial cells. Amniotic membrane denuding usually involves treatment with EDTA and/or proteolytic enzymes; in many cases additional mechanical scraping is required. Although ensuring limbal cell proliferation, these methods are not standardized, require relatively long treatment times and can result in membrane damage. We propose to use 0.5 M NaOH to reliably remove amniotic cells from the membrane. This method was used before to lyse cells for DNA isolation and radioactivity counting. Gently rubbing a cotton swab soaked in NaOH over the epithelial side of amniotic membrane leads to nearly complete and easy removal of adherent cells in less than a minute. The denuded membrane is subsequently washed in a neutral buffer. Cell removal was more thorough and uniform than with EDTA, or EDTA plus mechanical scraping with an electric toothbrush, or n-heptanol plus EDTA treatment. NaOH-denuded amniotic membrane did not show any perforations compared with mechanical or thermolysin denuding, and showed excellent preservation of immunoreactivity for major basement membrane components including laminin α2, γ1-γ3 chains, α1/α2 and α6 type IV collagen chains, fibronectin, nidogen-2, and perlecan. Sodium hydroxide treatment was efficient with fresh or cryopreserved (10% dimethyl sulfoxide or 50% glycerol amniotic membrane. The latter method is a common way of membrane storage for subsequent grafting in the European Union. NaOH-denuded amniotic membrane supported growth of human limbal epithelial cells, immortalized corneal epithelial cells, and induced pluripotent stem cells. This simple, fast and reliable method can be used to

  6. Amniotic membrane allografts: development and clinical utility in ophthalmology

    Directory of Open Access Journals (Sweden)

    Rizzuti A

    2014-12-01

    Full Text Available Allison Rizzuti,1,2 Adam Goldenberg,1 Douglas R Lazzaro1,2 1SUNY Downstate Medical Center, 2Kings County Hospital Center, Brooklyn, NY, USA Abstract: Amniotic membrane, the innermost layer of the placenta, is a tissue that promotes epithelialization, while decreasing inflammation, neovascularization, and scarring. It is used in the surgical management of a wide variety of ophthalmic conditions where it functions as a graft or patch in ocular surface reconstruction. The development of new preservation techniques, as well as a sutureless amniotic membrane, has allowed for easier, in-office placement, without the disadvantages of an operating room procedure. The purpose of this review is to describe the historical development of amniotic membrane in ophthalmology and to describe its current clinical applications, particularly focusing on recent advances. Keywords: ocular surface, cornea, stem cells, prokera, allograft, patch, transplantation

  7. 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.

  8. A altura uterina é capaz de diagnosticar os desvios do volume de líquido amniótico? Is uterine height able to diagnose amniotic fluid volume deviations?

    Directory of Open Access Journals (Sweden)

    Djacyr Magna Cabral Freire

    2013-02-01

    deveriam ter esse exame realizado.PURPOSE: To evaluate the performance of a Brazilian reference curve of fundal height (FH regarding its capacity of screening the deviations of volume of amniotic fluid using a Brazilian reference curve of amniotic fluid index (AFI as gold standard. METHODS: This was a cross-sectional study evaluating 753 pregnant women receiving prenatal care at the public health services of João Pessoa (PB, from March to October 2006, who had a routine ultrasound exam scheduled for after 26 weeks of gestational age. Cases with diagnoses of twin pregnancy, intrauterine fetal death and major fetal malformations were excluded. Besides socio-demographic information, data regarding fundal height measured in a standard way, estimated fetal weight, AFI and gestational age at the time of the ultrasound exam were also collected. The capacity of the FH curve to predict deviations of the amniotic fluid volume was assessed using the Brazilian curve of AFI according to gestational age as the gold standard. For this purpose, sensitivity, specificity, positive and negative predictive values were estimated for different cut-off points. RESULTS: The measurement of FH identified 10.5% of women as having low FH possibly associated with oligohydramnios and 25.2% as having high FH possibly associated with polyhydramnios. Using a Brazilian reference curve of AFI, the FH was able to poorly predict the occurrence of oligohydramnios (sensitivity ranging from 37 to 28% and to reasonably predict the occurrence of polyhydramnios (sensitivity ranging from 88 to 69%. CONCLUSIONS: The measurement of fundal height showed a poor performance for predicting oligohydramnios and a reasonable performance for predicting polyhydramnios. Its use for this purpose is then only supported in settings where the ultrasound exam is not easily or routinely available in order to help define priorities for cases that should have this exam performed.

  9. Diagnóstico do oligoâmnio pela ultra-sonografia: uso de diferentes medidas do maior bolsão comparadas ao ILA Diagnostic capacity of oligohydramnios by ultrasound using different measures of the maximum pool depth of amniotic fluid in comparison to AFI

    Directory of Open Access Journals (Sweden)

    Maria Regina Machado Perrotti

    2000-02-01

    Full Text Available Objetivo: comparar a capacidade de diagnosticar oligoâmnio pela ultra-sonografia por meio de diferentes valores do maior bolsão de líquido amniótico, em comparação ao índice de líquido amniótico (ILA, em gestantes normais, de 36 a 42 semanas de gestação. Métodos: realizou-se um estudo descritivo de validação de método diagnóstico, incluindo 875 gestantes normais. Mediante um exame ultra-sonográfico obstétrico de rotina, foi feita a medida do maior bolsão de líquido amniótico para o diagnóstico de oligoâmnio, utilizando como padrão-ouro o índice de líquido amniótico. Os dados foram analisados por meio do cálculo da sensibilidade e da especificidade da medida do maior bolsão de líquido amniótico, utilizando os diferentes pontos de corte de 10, 20 e 30 mm, em comparação aos valores normais do índice de líquido amniótico determinados pelos percentis 2,5 e 10 nas respectivas idades gestacionais. Resultados: a medida do maior bolsão de líquido amniótico apresenta baixa sensibilidade para diagnosticar oligoâmnio quando se adotam os pontos de corte 10 e 20 mm, e boa sensibilidade e especificidade quando se adota 30 mm, quando comparadas às medidas do índice de líquido amniótico nos percentis 2,5 e 10 da curva normal. A sensibilidade e especificidade da medida do maior bolsão são melhores, quando se adota o ponto de corte de 30 mm para diagnosticar oligoâmnio em comparação ao percentil 2,5. Conclusões: a capacidade de diagnosticar oligoâmnio pela medida do maior bolsão é satisfatória apenas com o ponto de corte em 30 mm.Purpose: to compare the capacity of diagnosing oligohy-dramnios by ultrasound using different measures of the maximum pool depth of amniotic fluid in comparison to the amniotic fluid index among normal pregnant women from the 36th to the 42nd week of gestation. Methods: a descriptive study of diagnostic validity was perfomed, on 875 normal pregnant women who were studied through

  10. Embolia pulmonar por líquido amniótico: relato de caso e revisão da literatura Pulmonary amniotic fluid embolism syndrome: case report and literature review

    Directory of Open Access Journals (Sweden)

    Edmilton Pereira de Almeida

    2007-06-01

    ório, hipotensão arterial, sintomatologia neurológica e coagulopatia que se iniciou durante ou em 30 min do trabalho de parto. O diagnóstico é de exclusão de outras afecções mais comuns, tais como eclâmpsia, sepse, miocardiopatia periparto, anafilaxia, tromboembolismo pulmonar, reações transfusionais, complicações anestésicas e estenose mitral, que podem surgir ou agravar-se durante o trabalho de parto. Pode ocorrer durante ou logo após aborto cirúrgico, amniocentese ou colocação de transdutor de pressão na cavidade uterina. O tratamento é o de suporte hemodinâmico, ventilatório e correção dos distúrbios de coagulação. A interrupção imediata da gestação, através de cesariana, parece trazer benefícios para a mãe e concepto. Os intensivistas e obstetras devem estar atentos para esta entidade com o intuito de diagnóstico e tratamento imediatos.BACKGROUND AND OBJECTIVES: Amniotic fluid embolism is a rare complication of pregnancy whose physiopathology is not completely known and still not frequently remembered by intensive care physicians and obstetricians. The main aim of this case report was to emphasize the need of better knowledge of this disease by physicians. CASE REPORT: A 15 years old, primigravida, 35th week of gestation was admitted in our intensive care unit (ICU with a sudden respiratory distress. Soon after admission she was intubated and put on mechanical ventilation. After stabilization she was transferred to the operating room and a cesarean section was undertaken. During the operation, instead of fluid reposition with crystalloids in large amounts, her hemodynamic status deteriorated and it was necessary to use vasoactive drugs. After three days on mechanical ventilation she was successfully weaned and discharged from the intensive care unit (ICU after 6 days. She and her baby were discharged from hospital on13th day of hospitalization. CONCLUSIONS: Amniotic fluid embolism is a rare and catastrophic complication of pregnancy, at

  11. 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

  12. Prevalence and Clinical Significance of Sterile Intra-amniotic Inflammation in Patients with Preterm Labor and Intact Membranes

    Science.gov (United States)

    Romero, Roberto; Miranda, Jezid; Chaiworapongsa, Tinnakorn; Korzeniewski, Steven J.; Chaemsaithong, Piya; Gotsch, Francesca; Dong, Zhong; Ahmed, Ahmed I.; Yoon, Bo Hyun; Hassan, Sonia S.; Kim, Chong J.; Yeo, Lami

    2014-01-01

    Problem Inflammation and infection play a major role in preterm birth. The purpose of this study was to: 1) determine the prevalence and clinical significance of sterile intra-amniotic inflammation; and 2) examine the relationship between amniotic fluid (AF) concentrations of high mobility group box-1 (HMGB1) and the interval from amniocentesis-to-delivery in patients with sterile intra-amniotic inflammation. Method of Study AF samples obtained from 135 women with preterm labor and intact membranes were analyzed using cultivation techniques as well as broad-range PCR and mass spectrometry (PCR/ESI-MS). Sterile intra-amniotic inflammation was defined when patients with negative AF cultures and without evidence of microbial footprints had intra-amniotic inflammation (AF interleukin-6 ≥ 2.6 ng/mL). Results 1) The frequency of sterile intra-amniotic inflammation was significantly greater than that of microbial-associated intra-amniotic inflammation [26% (35/135) vs. 11% (15/135); (p=0.005)]; 2) patients with sterile intra-amniotic inflammation delivered at comparable gestational ages, had similar rates of acute placental inflammation and adverse neonatal outcomes as patients with microbial-associated intra-amniotic inflammation; and 3) patients with sterile intra-amniotic inflammation and high AF concentrations of HMGB1 (≥ 8.55 ng/mL) delivered earlier than those with low AF concentrations of HMGB1 (p=0.02). Conclusions 1) sterile intra-amniotic inflammation is more frequent than microbial-associated intra-amniotic inflammation; and 2) we propose that danger signals participate in sterile intra-amniotic inflammation in the setting of preterm labor. PMID:25078709

  13. 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.

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    Chan-Wook Park

    Full Text Available 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 <23 ng/ml. We examined the association between amniocentesis-to-delivery interval and inflammatory status in the extra-placental 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; P<0.05. Amniocentesis-to-delivery interval was an independent predictor of the development of both chorio-deciduitis and amnionitis after correction for confounding variables such as gestational age at delivery in the setting of PTL, but not preterm-PROM.These data confirm for the first time that, in cases of both PTL and preterm-PROM with sterile AF, more time is required to develop chorio-deciduitis with amnionitis than chorio-deciduitis alone in extra-placental membranes. Moreover

  14. Relationships between sex hormones assessed in amniotic fluid, and maternal and umbilical cord serum: What is the best source of information to investigate the effects of fetal hormonal exposure?

    NARCIS (Netherlands)

    van de Beek, C.; Thijssen, J.H.H.; Kettenis, P.T.; van Goozen, S.H.; Buitelaar, J.K.

    2004-01-01

    Levels of testosterone (T) (total and free), androstenedione (A4), dehydroepiandrosterone sulphate (DHEAS), sex hormone-binding globulin (SHBG), and estradiol (E2) were measured by radioimmunoassay (RIA) in 156 normal pregnancies (77 male and 79 female fetuses). Samples were obtained from amniotic

  15. Relationships between sex hormones assessed in amniotic fluid, and maternal and umbilical cord serum: what is the best source of information to investigate the effects of fetal hormonal exposure?

    NARCIS (Netherlands)

    Beek, C.; Thijssen, J.H.; Cohen-Kettenis, P.T.; Goozen, S.H. van

    2004-01-01

    Levels of testosterone (T) (total and free), androstenedione (A4), dehydroepiandrosterone sulphate (DHEAS), sex hormone-binding globulin (SHBG), and estradiol (E2) were measured by radioimmunoassay (RIA) in 156 normal pregnancies (77 male and 79 female fetuses). Samples were obtained from amniotic

  16. Experimental and numerical investigations of fluid flow for optimized in vitro stem cell loading in xenografts

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    Ott Robert

    2017-09-01

    Full Text Available In dentofacial surgery, augmentation procedures employing xenografts have become a reliable treatment. Recent studies, however, have shown significant enhance-ments of the in vivo bone tissue augmentation using mesenchymal stem cells loaded into bone grafts. We conducted experimental and numerical investigations in flow perfusion systems to determine flow conditions which allow for homogenous stem cell distribution in BioOss Block (Geistlich Pharma AG, Switzerland xenografts. Pressure gradient-velocity characteristics and flow distributions were investigated experimentally and numerically at steady state flow conditions with Reynolds numbers (Re ranging from 0.01 ≤ Re ≤ 0.40. Distilled water at 20°C with a dynamic viscosity of 1.002 mPa.s and a density of 998 kg/m3 was used. The geometry utilized in three-dimensional computa-tional fluid dynamics (CFD simulation was obtained by means of micro-computed tomography (μCT. Results of CFD analysis are in good accordance with experimental data. The comparison of the pressure gradient-velocity characteris-tics for experimental and numerical data yields a relative error of 3.6%. According to Darcy’s law for creeping fluid flow the experimentally determined permeability is 2.55.10-9 m2. Moreover, numerical flow distribution analysis shows an increasingly heterogenic streamline distribution for increasing Reynolds numbers. Experimentally validated CFD simulations introduced in this study provide a tool to assess optimal flow conditions for a homogenous stem cell distribution in perfusion flow systems.

  17. Avaliação da maturidade pulmonar fetal pela contagem dos corpos lamelares no líquido amniótico Evaluation of fetal lung maturity by lamellar bodies counting in amniotic fluid

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    Beatriz Maykot Kuerten Gil

    2010-03-01

    Full Text Available OBJETIVO: comparar o teste de contagem de corpos lamelares (CCL no líquido amniótico com o teste da polarização fluorescente (PF como parâmetro diagnóstico para avaliação da maturidade pulmonar fetal. MÉTODO: estudo transversal, analítico e controlado realizado com 60 gestantes atendidas no período de março de 2002 a dezembro de 2007. Foram colhidas amostras de líquido amniótico e realizados os testes de CCL e PF (TDxFLM II, considerados de referência, e comparados à presença ou ausência da Síndrome do Desconforto Respiratório (SDR. Foram estabelecidos valores de corte para maturidade de 30 mil corpos lamelares/µL para o teste da CCL e 55 mg/g de albumina para o PF. Foram avaliadas as características maternas e perinatais, a evolução neonatal e o desempenho dos testes diagnósticos para predição da maturidade pulmonar fetal. Na análise estatística, foram utilizadas medidas descritivas e calculados os valores referentes à sensibilidade, especificidade, valor preditivo positivo e negativo dos testes, considerando-se significativos valores de pPURPOSE: to compare the lamellar body number density (LBND count in amniotic fluid using the fluorescent polarization (FP test as a diagnostic parameter for the assessment of fetal pulmonary maturity. METHOD: this was an analytical, controlled cross-sectional study conducted on 60 pregnant women from March 2002 to December 2007. Amniotic fluid specimens were obtained by amniocentesis or at the time of caesarean section, and submitted to the LBND and FP tests (TDxFLM®, Abbott Laboratories, the latter considered to be a reference test, and compared in terms of the presence or absence of respiratory distress syndrome (RDS. Cut-off values for maturity were established at 30,000 lamellar bodies/µL for the LBND test and 55 mg/g albumin for the FP test. Maternal and perinatal characteristics and neonatal evolution were evaluated, and the performance of the diagnostic tests regarding

  18. Human amniotic epithelial cells combined with silk fibroin scaffold in the repair of spinal cord injury

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    Ting-gang Wang

    2016-01-01

    Full Text Available Treatment and functional reconstruction after central nervous system injury is a major medical and social challenge. An increasing number of researchers are attempting to use neural stem cells combined with artificial scaffold materials, such as fibroin, for nerve repair. However, such approaches are challenged by ethical and practical issues. Amniotic tissue, a clinical waste product, is abundant, and amniotic epithelial cells are pluripotent, have low immunogenicity, and are not the subject of ethical debate. We hypothesized that amniotic epithelial cells combined with silk fibroin scaffolds would be conducive to the repair of spinal cord injury. To test this, we isolated and cultured amniotic epithelial cells, and constructed complexes of these cells and silk fibroin scaffolds. Implantation of the cell-scaffold complex into a rat model of spinal cord injury resulted in a smaller glial scar in the damaged cord tissue than in model rats that received a blank scaffold, or amniotic epithelial cells alone. In addition to a milder local immunological reaction, the rats showed less inflammatory cell infiltration at the transplant site, milder host-versus-graft reaction, and a marked improvement in motor function. These findings confirm that the transplantation of amniotic epithelial cells combined with silk fibroin scaffold can promote the repair of spinal cord injury. Silk fibroin scaffold can provide a good nerve regeneration microenvironment for amniotic epithelial cells.

  19. Donor mesenchymal stem cells home to maternal wounds after transamniotic stem cell therapy (TRASCET) in a rodent model.

    Science.gov (United States)

    Graham, Christopher D; Shieh, Hester F; Brazzo, Joseph A; Zurakowski, David; Fauza, Dario O

    2017-06-01

    Transamniotic stem cell therapy (TRASCET) with amniotic fluid-derived MSCs (afMSCs) has emerged experimentally as a practical treatment strategy for congenital anomalies. In this study, we sought to determine whether afMSCs migrate to the mother following TRASCET. Pregnant rat dams were divided into three groups. Two groups received volume-matched injections into all amniotic cavities of either a suspension of afMSCs labeled with a luciferase reporter gene or the luciferase protein alone. In a third group, a suspension of labeled cells was aliquoted onto the serosal surface of the uterus. Maternal samples from the laparotomy scar (fascia and skin separately), bone marrow, and peripheral blood were procured, along with placenta and umbilical cord. Specimens were screened for luminescence via microplate luminometry. Luminescence was detected in 60% (9/15) of the fascial scars from the group receiving intraamniotic injection of afMSCs, but in none of the other groups (Pcells in the placenta and their presence in maternal fascia (Wald test=10.2; P=0.001). Amniotic mesenchymal stem cells migrate to maternal sites of injury after intraamniotic injection. Maternal homing of donor cells must be considered in the setting of transamniotic stem cell therapy. N/A (animal and laboratory study). Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Prenatal toxoplasmosis diagnosis from amniotic fluid by PCR Diagnóstico pré-natal da toxoplasmose no líquido amniótico através da técnica de PCR

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    Paula Vieira Teixeira Vidigal

    2002-02-01

    Full Text Available Toxoplasmosis is one of the most common infections all over the world. Most cases are asymptomatic, except in immunosuppressed individuals and fetuses, which can be seriously damaged. Prenatal diagnosis should be made as soon as possible since treatment of the mother can minimize fetal sequelae. Our aim in this study was to test the polymerase chain reaction technique (PCR in 86 samples of amniotic fluid from women who seroconverted during pregnancy. DNA was amplified using external primers and, in a second step, internal primers, in a nested PCR system. Samples were also inoculated into mice and the newborn were evaluated by T. gondii serology, skull x-ray, transfontanel ultrasound, fundoscopic examination, lumbar puncture and clinical examination. PCR was positive in seven cases and negative in 79. Among PCR-positive cases, two were negative by inoculation into mice and by clinical evaluation; among PCR-negative ones, three had clinical evidence of toxoplasmosis and one was positive after inoculation into mice. PCR showed values of sensitivity = 62.5% and specificity = 97.4%; the values of inoculation into mice where 42.9% and 100%, respectively. Although PCR should not be used alone for prenatal diagnosis of congenital toxoplasmosis, it is a promising method and deserves more studies to improve its efficacy.A toxoplasmose é infecção freqüente em todo o mundo, mas na maioria dos casos não traz repercussões importantes para o paciente, exceto indivíduos imunodeprimidos e fetos, os quais podem apresentar graves seqüelas. O diagnóstico precoce durante a gravidez é altamente desejável, já que o tratamento da gestante reduz a freqüência e gravidade da infecção fetal. Neste estudo aplicou-se a técnica de PCR em 86 amostras de líquido amniótico de gestantes que apresentaram soroconversão durante a gravidez. O DNA foi amplificado usando-se iniciadores externos e internos, num sistema de nested PCR. As amostras foram tamb

  1. Regenerative medicine for the kidney: renotropic factors, renal stem/progenitor cells, and stem cell therapy.

    Science.gov (United States)

    Maeshima, Akito; Nakasatomi, Masao; Nojima, Yoshihisa

    2014-01-01

    The kidney has the capacity for regeneration and repair after a variety of insults. Over the past few decades, factors that promote repair of the injured kidney have been extensively investigated. By using kidney injury animal models, the role of intrinsic and extrinsic growth factors, transcription factors, and extracellular matrix in this process has been examined. The identification of renal stem cells in the adult kidney as well as in the embryonic kidney is an active area of research. Cell populations expressing putative stem cell markers or possessing stem cell properties have been found in the tubules, interstitium, and glomeruli of the normal kidney. Cell therapies with bone marrow-derived hematopoietic stem cells, mesenchymal stem cells, endothelial progenitor cells, and amniotic fluid-derived stem cells have been highly effective for the treatment of acute or chronic renal failure in animals. Embryonic stem cells and induced pluripotent stem cells are also utilized for the construction of artificial kidneys or renal components. In this review, we highlight the advances in regenerative medicine for the kidney from the perspective of renotropic factors, renal stem/progenitor cells, and stem cell therapies and discuss the issues to be solved to realize regenerative therapy for kidney diseases in humans.

  2. Acceleration of Regeneration of Large-Gap Peripheral Nerve Injuries Using Acellular Nerve Allografts Plus Amniotic Fluid Derived Stem Cells (AFS)

    Science.gov (United States)

    2017-09-01

    Elsaidi G, Ritting A, Hick J, Tan KA, Smith TL, Smith BP, Koman LA. Gene expression of myogenic regulatory factors, nicotinic acetylcholine receptor...804-809. 3. Ryall JG, Plant DR, Gregorevic P, Sillence MN, Lynch GS. Beta-2 agonist administration reverses muscle wasting and improves muscle

  3. Cervical length and gestational age at admission as predictors of intra-amniotic inflammation in preterm labor with intact membranes.

    Science.gov (United States)

    Palacio, M; Cobo, T; Bosch, J; Filella, X; Navarro-Sastre, A; Ribes, A; Gratacós, E

    2009-10-01

    To evaluate cervical length and gestational age as predictors of intra-amniotic inflammation in patients admitted because of preterm labor and intact membranes. Ninety-three pregnant women with preterm labor and intact membranes were included in our study. Cervical length was measured on admission by transvaginal sonography and transabdominal amniocentesis was performed within the first 48 h following admission. Positive amniotic fluid cultures defined intra-amniotic infection. Levels of intra-amniotic interleukin-6 (IL-6) were measured, and a receiver-operating characteristics (ROC) curve was constructed to determine the best cut-off point of IL-6 for predicting intra-amniotic infection. This value was then used as a basis for determining a cut-off of IL-6 for defining intra-amniotic inflammation. Considering inflammatory status, perinatal outcomes were evaluated and compared. Logistic regression was used to investigate associations of different explanatory variables with inflammatory status. A non-invasive approach for detection of intra-amniotic inflammation in women admitted because of preterm labor with intact membranes was evaluated. Intra-amniotic infection and inflammation rates were 14% and 28%, respectively. ROC curve analysis showed that the best cut-off value for IL-6 was 13.4 ng/mL for predicting intra-amniotic infection, which was comparable to the cut-off of 11.3 ng/mL reported previously by other authors (which we used to define inflammation). Regardless of the intra-amniotic microbial status, perinatal outcomes in women who developed intra-amniotic inflammation were worse than in those who did not. Cervical length or= 28 and membranes.

  4. Mesenchymal Stem Cells Obtained from Synovial Fluid Mesenchymal Stem Cell-Derived Induced Pluripotent Stem Cells on a Matrigel Coating Exhibited Enhanced Proliferation and Differentiation Potential.

    Science.gov (United States)

    Zheng, Yu-Liang; Sun, Yang-Peng; Zhang, Hong; Liu, Wen-Jing; Jiang, Rui; Li, Wen-Yu; Zheng, You-Hua; Zhang, Zhi-Guang

    2015-01-01

    Induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs) serve as a promising source for cell-based therapies in regenerative medicine. However, optimal methods for transforming iPSCs into MSCs and the characteristics of iPSC-MSCs obtained from different methods remain poorly understood. In this study, we developed a one-step method for obtaining iPSC-MSCs (CD146+STRO-1+ MSCs) from human synovial fluid MSC-derived induced iPSCs (SFMSC-iPSCs). CD146-STRO-1-SFMSCs were reprogrammed into iPSCs by transduction with lentivirus-mediated Sox2, Oct-3/4, klf4, and c-Myc. SFMSC-iPSCs were maintained with mTeSR1 medium in Matrigel-coated culture plates. Single dissociated cells were obtained by digesting the SFMSC-iPSCs with trypsin. The dissociated cells were then plated into Matrigel-coated culture plate with alpha minimum essential medium supplemented with 10% fetal bovine serum, 1× Glutamax, and the ROCK inhibitor Y-27632. Cells were then passaged in standard cell culture plates with alpha minimum essential medium supplemented with 10% fetal bovine serum and 1× Glutamax. After passaging in vitro, the cells showed a homogenous spindle-shape similar to their ancestor cells (SFMSCs), but with more robust proliferative activity. Flow cytometric analysis revealed typical MSC surface markers, including expression of CD73, CD90, CD105, and CD44 and lack of CD45, CD34, CD11b, CD19, and HLA-DR. However, these cells were positive for CD146 and stro-1, which the ancestor cells were not. Moreover, the cells could also be induced to differentiate in osteogenic, chondrogenic, and adipogenic lineages in vitro. The differentiation potential was improved compared with the ancestor cells in vitro. The cells were not found to exhibit oncogenicity in vivo. Therefore, the method presented herein facilitated the generation of STRO-1+CD146+ MSCs from SFMSC-iPSCs exhibiting enhanced proliferation and differentiation potential.

  5. Mesenchymal Stem Cells Obtained from Synovial Fluid Mesenchymal Stem Cell-Derived Induced Pluripotent Stem Cells on a Matrigel Coating Exhibited Enhanced Proliferation and Differentiation Potential.

    Directory of Open Access Journals (Sweden)

    Yu-Liang Zheng

    Full Text Available Induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs serve as a promising source for cell-based therapies in regenerative medicine. However, optimal methods for transforming iPSCs into MSCs and the characteristics of iPSC-MSCs obtained from different methods remain poorly understood. In this study, we developed a one-step method for obtaining iPSC-MSCs (CD146+STRO-1+ MSCs from human synovial fluid MSC-derived induced iPSCs (SFMSC-iPSCs. CD146-STRO-1-SFMSCs were reprogrammed into iPSCs by transduction with lentivirus-mediated Sox2, Oct-3/4, klf4, and c-Myc. SFMSC-iPSCs were maintained with mTeSR1 medium in Matrigel-coated culture plates. Single dissociated cells were obtained by digesting the SFMSC-iPSCs with trypsin. The dissociated cells were then plated into Matrigel-coated culture plate with alpha minimum essential medium supplemented with 10% fetal bovine serum, 1× Glutamax, and the ROCK inhibitor Y-27632. Cells were then passaged in standard cell culture plates with alpha minimum essential medium supplemented with 10% fetal bovine serum and 1× Glutamax. After passaging in vitro, the cells showed a homogenous spindle-shape similar to their ancestor cells (SFMSCs, but with more robust proliferative activity. Flow cytometric analysis revealed typical MSC surface markers, including expression of CD73, CD90, CD105, and CD44 and lack of CD45, CD34, CD11b, CD19, and HLA-DR. However, these cells were positive for CD146 and stro-1, which the ancestor cells were not. Moreover, the cells could also be induced to differentiate in osteogenic, chondrogenic, and adipogenic lineages in vitro. The differentiation potential was improved compared with the ancestor cells in vitro. The cells were not found to exhibit oncogenicity in vivo. Therefore, the method presented herein facilitated the generation of STRO-1+CD146+ MSCs from SFMSC-iPSCs exhibiting enhanced proliferation and differentiation potential.

  6. Fatores maternos e neonatais associados ao mecônio no líquido amniótico em um centro de parto normal Factores maternos y neonatales asociados al meconio en el líquido amniótico en un centro de parto normal Meconium-stained amniotic fluid and maternal and neonatal factors associated

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    Ruth Hitomi Osava

    2012-12-01

    95% 1,60;2,10, cesárea (RP = 2,65; IC95% 2,17;3,24 e índice de Apgar OBJECTIVE: To identify the frequency and maternal and neonatal factors associated with meconium-stained amniotic fluid at birth. METHODS: Cross-sectional study carried out with 2,441 births at an in-hospital birth center in the city of São Paulo (Southeastern Brazil in March and April, 2005. The association between meconium-stained amniotic fluid and the independent variables (maternal age, parity, previous c-section or not, gestational age, obstetric history, oxytocin use in the labor, cervical dilation at admission, mode of current delivery, newborn weight, Apgar score at the 1st and 5th minute was expressed as prevalence ratio (PR. RESULTS: Meconium-stained amniotic fluid was verified in 11.9% of the births; 68.2% of these were normal births and 38.8% c-sections. Meconium was associated with: primiparity (PR=1.49, 95%CI 1.29; 1.73, gestational age ≥ 41 weeks (PR = 5.05, 95%CI 1.93;13.25, oxytocin in labor (PR = 1.83, 95%CI 1.60; 2.10, c- section (PR = 2.65, 95%CI 2.17; 3.24 and Apgar scores < 7 at the 5th minute (PR = 2.96, 95%CI 2,94;2,99. Neonatal mortality was 1.6/1,000 live births. Meconium-stained amniotic fluid was found in 50% of neonatal deaths and it was associated with higher rates of surgical deliveries. CONCLUSIONS: Oxytocin use, worse conditions of the newborn after the delivery and increased c-section rates were factors associated with meconium-stained amniotic fluid. Routine use of oxytocin in the intrapartum period could be evaluated due to its association with meconium-stained amniotic fluid.

  7. AMNIOTIC MEMBRANE TRANSPLANTATION FOR KERATITIS.

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    Snezhana Murgova

    2015-06-01

    Full Text Available Keratitis without proper management tends to perforate the cornea, resulting in severe adverse consequences. In recent studies, amniotic membrane is reported to have anti-inflammatory effect and promote wound healing of corneal ulcer. Purpose: To report on the efficacy of permanent amniotic membrane transplantation (AMT in the treatment of keratitis. Case report: A 58-year-old man with severe keratitis in both eyes caused by long term administration of topical anesthetic (alcaine for electric ophthalmia. Single layer of amniotic membrane (AM was placed on the defect and secured to the limbus with interrupted 10-0 nylon sutures. A bandage contact lens was applied on the AM. Postoperative medication included topical antibiotic, artificial tears and mydriatic. Three months later corticosteroid was included. There was an immediate decrease of patient’s pain after surgery. Complete epithelialization was noted after 1 month. Conclusion: AMT is an alternative adjunctive method of treatment of keratitis; it promotes epithelialization process, decreased inflammation, corneal haze and neovascularization.

  8. Different Tissue-Derived Stem Cells: A Comparison of Neural Differentiation Capability

    Science.gov (United States)

    Bonaventura, Gabriele; Chamayou, Sandrine; Liprino, Annalisa; Guglielmino, Antonino; Fichera, Michele; Caruso, Massimo; Barcellona, Maria Luisa

    2015-01-01

    Background Stem cells are capable of self-renewal and differentiation into a wide range of cell types with multiple clinical and therapeutic applications. Stem cells are providing hope for many diseases that currently lack effective therapeutic methods, including strokes, Huntington's disease, Alzheimer's and Parkinson's disease. However, the paucity of suitable cell types for cell replacement therapy in patients suffering from neurological disorders has hampered the development of this promising therapeutic approach. Aim The innovative aspect of this study has been to evaluate the neural differentiation capability of different tissue-derived stem cells coming from different tissue sources such as bone marrow, umbilical cord blood, human endometrium and amniotic fluid, cultured under the same supplemented media neuro-transcription factor conditions, testing the expression of neural markers such as GFAP, Nestin and Neurofilaments using the immunofluorescence staining assay and some typical clusters of differentiation such as CD34, CD90, CD105 and CD133 by using the cytofluorimetric test assay. Results Amniotic fluid derived stem cells showed a more primitive phenotype compared to the differentiating potential demonstrated by the other stem cell sources, representing a realistic possibility in the field of regenerative cell therapy suitable for neurodegenerative diseases. PMID:26517263

  9. Different Tissue-Derived Stem Cells: A Comparison of Neural Differentiation Capability.

    Directory of Open Access Journals (Sweden)

    Gabriele Bonaventura

    Full Text Available Stem cells are capable of self-renewal and differentiation into a wide range of cell types with multiple clinical and therapeutic applications. Stem cells are providing hope for many diseases that currently lack effective therapeutic methods, including strokes, Huntington's disease, Alzheimer's and Parkinson's disease. However, the paucity of suitable cell types for cell replacement therapy in patients suffering from neurological disorders has hampered the development of this promising therapeutic approach.The innovative aspect of this study has been to evaluate the neural differentiation capability of different tissue-derived stem cells coming from different tissue sources such as bone marrow, umbilical cord blood, human endometrium and amniotic fluid, cultured under the same supplemented media neuro-transcription factor conditions, testing the expression of neural markers such as GFAP, Nestin and Neurofilaments using the immunofluorescence staining assay and some typical clusters of differentiation such as CD34, CD90, CD105 and CD133 by using the cytofluorimetric test assay.Amniotic fluid derived stem cells showed a more primitive phenotype compared to the differentiating potential demonstrated by the other stem cell sources, representing a realistic possibility in the field of regenerative cell therapy suitable for neurodegenerative diseases.

  10. Osteogenic differentiation of amniotic epithelial cells: synergism of pulsed electromagnetic field and biochemical stimuli

    OpenAIRE

    Wang, Qian; Wu, Wenchao; Han, Xiaoyu; Zheng, Ai; Lei, Song; Wu, Jiang; Chen, Huaiqing; He, Chengqi; Luo, Fengming; Liu, Xiaojing

    2014-01-01

    Background Pulsed electromagnetic field (PEMF) is a non-invasive physical therapy used in the treatment of fracture nonunion or delayed healing. PEMF can facilitate the osteogenic differentiation of bone marrow mesenchymal stem cells in vitro. Amniotic epithelial cells (AECs) have been proposed as a potential source of stem cells for cell therapy. However, whether PEMF could modulate the osteogenic differentiation of AECs is unknown. In the present study, the effects of PEMF on the osteogenic...

  11. Líquido amniótico, atividade física e imersão em água na gestação Amniotic fluid, physical activity and water immersion during pregnancy

    Directory of Open Access Journals (Sweden)

    Márcia San Juan Dertkigil

    2005-12-01

    Full Text Available O objetivo desta revisão foi avaliar a inter-relação entre volume do líquido amniótico, atividade física e imersão em água na gestação. Utilizando diferentes combinações desses termos, foram consultadas as bases de dados do MedLine e do Scielo, no período de 1980 a 2005 para a identificação de artigos relevantes ao assunto. O líquido amniótico é fundamental no desenvolvimento dos sistemas músculo-esquelético, gastrointestinal e respiratório fetal. Sua avaliação no passado era imprecisa, mas com o advento da ultra-sonografia, tornou-se fácil e não invasiva. Surgiram técnicas para a aferição do seu volume e curvas de normalidade. Mais recentemente, têm-se tentado diversas técnicas para a correção do seu volume, na tentativa de diminuir a morbidade associada. Um método não invasivo muito utilizado no Brasil é a hiperhidratação materna. Outros autores sugerem medidas como a imersão estática ou com atividade física em água, muito na moda nos dias de hoje. Ainda são poucos os estudos relativos à prática da atividade física na água para gestantes e seus benefícios maternos, fetais e perinatais. A presente revisão visa aprofundar os conhecimentos acerca dos exercícios aeróbicos sob imersão de gestantes em água, no que diz respeito ao volume de líquido amniótico e bem-estar fetal durante a gestação.The objective of this paper was to assess the relationship between amniotic liquid volume, physical activity and immersion in water during pregnancy. By using different combinations of these terms MedLine and Scielo databases were consulted in the period of 1980 and 2005 for subject related articles. Amniotic liquid is fundamental for fetal muscle skeletal, gastrointestinal and respiratory systems development. In the past amniotic liquid evaluation was poor but with the introduction of ultrasonography it has become easy and non-invasive. New techniques to measure volume and normality curves emerged. More

  12. Clinical chorioamnionitis at term VIII: a rapid MMP-8 test for the identification of intra-amniotic inflammation.

    Science.gov (United States)

    Chaiyasit, Noppadol; Romero, Roberto; Chaemsaithong, Piya; Docheva, Nikolina; Bhatti, Gaurav; Kusanovic, Juan Pedro; Dong, Zhong; Yeo, Lami; Pacora, Percy; Hassan, Sonia S; Erez, Offer

    2017-07-26

    Clinical chorioamnionitis is the most common infection/inflammatory process diagnosed in labor and delivery units worldwide. The condition is a syndrome that can be caused by (1) intra-amniotic infection, (2) intra-amniotic inflammation without demonstrable microorganisms (i.e. sterile intra-amniotic inflammation), and (3) maternal systemic inflammation that is not associated with intra-amniotic inflammation. The presence of intra-amniotic inflammation is a risk factor for adverse maternal and neonatal outcomes in a broad range of obstetrical syndromes that includes clinical chorioamnionitis at term. Although the diagnosis of intra-amniotic infection has relied on culture results, such information is not immediately available for patient management. Therefore, the diagnosis of intra-amniotic inflammation could be helpful as a proxy for intra-amniotic infection, while results of microbiologic studies are pending. A rapid test is now available for the diagnosis of intra-amniotic inflammation, based on the determination of neutrophil collagenase or matrix metalloproteinase-8 (MMP-8). The objectives of this study were (1) to evaluate the diagnostic indices of a rapid MMP-8 test for the identification of intra-amniotic inflammation/infection in patients with the diagnosis of clinical chorioamnionitis at term, and (2) to compare the diagnostic performance of a rapid MMP-8 test to that of a conventional enzyme-linked immunosorbent assay (ELISA) interleukin (IL)-6 test for patients with clinical chorioamnionitis at term. A retrospective cohort study was conducted. A transabdominal amniocentesis was performed in patients with clinical chorioamnionitis at term (n=44). Amniotic fluid was analyzed using cultivation techniques (for aerobic and anaerobic bacteria as well as genital Mycoplasmas) and broad-range polymerase chain reaction (PCR) coupled with electrospray ionization mass spectrometry (PCR/ESI-MS). Amniotic fluid IL-6 concentrations were determined by ELISA, and rapid

  13. Methods of amniotic membrane fixation in ocular amniotic membrane surgeries

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    Shu-Rong Wang

    2016-05-01

    Full Text Available Various ocular surface disorders like alkali burns and corneal ulcers can all cause damage to the cornea and conjunctiva, and often induce corneal neovascularization(CNVthat affects the visual function. However, amniotic membranes(AMcan promote the rapid epithelization of acute injured corneas and conjunctiva defects, diminish scarring, and perform anti-inflammatory effect. Therefore, AM has been widely used in ocular surface reconstructions and treatment of CNV. But the key problem is how to fix the AM. Only ensuring the adhesive time and cover area with convenient operation and little stimulation can achieve the best curative effect. This article reviews the methods of AM fixation in AM patch technique.

  14. Prenatal diagnosis of amniotic band syndrome

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    Laxmi Devi Padmanabhan

    2016-01-01

    Full Text Available Amniotic band can cause a broad spectrum of anomalies ranging from simple band constrictions to major craniofacial and visceral defects. It can cause significant neonatal morbidity. Accurate diagnosis will help in the management of the present pregnancy and in counseling with regard to future pregnancies. Here we report three cases of amniotic band syndrome detected in the prenatal period.

  15. Periodontal disease and intra-amniotic complications in women with preterm prelabor rupture of membranes.

    Science.gov (United States)

    Radochova, Vladimira; Kacerovska Musilova, Ivana; Stepan, Martin; Vescicik, Peter; Slezak, Radovan; Jacobsson, Bo; Kacerovsky, Marian

    2017-08-04

    Periodontal disease is frequently suggested as a possible causal factor for preterm delivery. The link between periodontal disease and preterm delivery is a possible translocation of periopathogenic bacteria to the placenta and amniotic fluid as well as a systemic response to this chronic inflammatory disease. However, there is a lack of information on whether there is an association between clinical periodontal status in women with preterm prelabor rupture of membranes (PPROM) and the presence of microbial invasion of the amniotic cavity (MIAC) and intra-amniotic inflammation (IAI). Therefore, the main aim of this study was to evaluate the incidence and severity of periodontal disease in women with PPROM. The secondary aim was to characterize an association between periodontal status and the presence of intra-amniotic PPROM complications (MIAC and/or IAI). Seventy-eight women with PPROM at gestational ages between 24 + 0 and 36 + 6 weeks were included in this study. The samples of amniotic fluid were obtained at admission via transabdominal amniocentesis, and amniotic fluid interleukin (IL)-6 concentrations were determined using a point-of-care test. All women had a full-mouth recording to determine the periodontal and oral hygiene status. Probing pocket depth and clinical attachment loss were measured at four sites on each fully erupted tooth. In total, 45% (35/78) of women with PPROM had periodontal disease. Mild, moderate, and severe periodontal disease was present in 19% (15/78), 19% (15/78), and 6% (5/78) of women, respectively. The presence of MIAC and IAI was found in 28% (22/78) and 26% (20/78) of women, respectively. Periopathogenic bacteria (2 × Streptococcus intermedius and 1 × Fusobacterium nucleatum) was found in the amniotic fluid of 4% (3/78) of women. There were no differences in periodontal status between women with MIAC and/or IAI and women without these intra-amniotic complications. The presence of MIAC and IAI was not related

  16. Ethical issues in stem cell research and therapy.

    Science.gov (United States)

    King, Nancy Mp; Perrin, Jacob

    2014-07-07

    Rapid progress in biotechnology has introduced a host of pressing ethical and policy issues pertaining to stem cell research. In this review, we provide an overview of the most significant issues with which the stem cell research community should be familiar. We draw on a sample of the bioethics and scientific literatures to address issues that are specific to stem cell research and therapy, as well as issues that are important for stem cell research and therapy but also for translational research in related fields, and issues that apply to all clinical research and therapy. Although debate about the moral status of the embryo in human embryonic stem cell research continues to have relevance, the discovery of other highly multipotent stem cell types and alternative methods of isolating and creating highly multipotent stem cells has raised new questions and concerns. Induced pluripotent stem cells hold great promise, but care is needed to ensure their safety in translational clinical trials, despite the temptation to move quickly from bench to bedside. A variety of highly multipotent stem cells - such as mesenchymal stem/stromal cells and stem cells derived from amniotic fluid, umbilical cord blood, adipose tissue, or urine - present the opportunity for widespread biobanking and increased access. With these increased opportunities, however, come pressing policy issues of consent, control, and justice. The imperatives to minimize risks of harm, obtain informed consent, reduce the likelihood of the therapeutic misconception, and facilitate sound translation from bench to bedside are not unique to stem cell research; their application to stem cell research and therapy nonetheless merits particular attention. Because stem cell research is both scientifically promising and ethically challenging, both the application of existing ethical frameworks and careful consideration of new ethical implications are necessary as this broad and diverse field moves forward.

  17. Volume do líquido amniótico associado às anomalias fetais diagnosticadas em um centro de referência do nordeste brasileiro Amniotic fluid volume associated with fetal anomalies diagnosed in a reference center in the Brazilian Northeast

    Directory of Open Access Journals (Sweden)

    Carlos Noronha Neto

    2009-04-01

    Full Text Available OBJETIVO: determinar fatores associados ao volume de líquido amniótico e frequências de anomalias fetais em um centro de referência. MÉTODOS: realizou-se um estudo de corte transversal, com gestantes de risco, avaliadas pela ultrassonografia morfológica, no período de março de 2002 a março de 2006, em uma instituição em Recife (PE Brasil. O diagnóstico intraútero foi confirmado no pós-parto. As características sociodemográficas e obstétricas, o índice de líquido amniótico e a presença de anomalias fetais foram variáveis estudadas. Para verificar associação entre variáveis, foram utilizados testes χ2, exato de Fisher e t de Student, a um nível de significância de 5%. Foram calculados a razão de prevalência e o intervalo de confiança a 95%. Análise de regressão logística múltipla foi realizada, a um nível de significância de 5%. RESULTADOS: foram incluídas no estudo 257 (56,2% gestantes com anomalias congênitas e 200 sem anomalias confirmadas no pós-natal. As médias das idades maternas e gestacionais do parto foram 24,8±6,5 anos e 35,9±3,7 semanas, respectivamente. As anomalias fetais foram mais encontradas no sistema nervoso central (50,6% e trato geniturinário (23,0%. A presença de anomalias congênitas esteve associada significativamente ao líquido diminuído/oligohidrâmnio (p=0,0002 e líquido aumentado/polihidrâmnio (pPURPOSE: to determine factors associated to amniotic fluid volume and frequencies of fetal anomalies, in a reference center in Pernambuco. METHODS: a transversal study performed in high-risk pregnant women submitted to obstetrical morphological ultrasound, from March 2002 to March 2006, at an institution from Recife, Pernambuco, Brasil. The intrauterine diagnosis was confirmed after birth. Sociodemographic and obstetrical characteristics, amniotic liquid volume and presence of fetal anomalies were the variables studied. Fisher's exact, χ2, and Student's t tests, at a significance

  18. Transplante de células-tronco epiteliais límbicas alógenas expandidas ex vivo sobre membrana amniótica: relato de caso Transplantation of allogenic limbal epithelial stem cells cultivated ex vivo on amniotic membrane: case report

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    José Álvaro Pereira Gomes

    2009-04-01

    Full Text Available Paciente apresentou falência de transplante de limbo e conjuntiva de doador vivo alógeno no olho direito após ceratoconjuntivite epidêmica. Após alguns meses, foi submetida a transplante de células-tronco epiteliais límbicas alógenas cultivadas ex vivo sobre membrana amniótica (primeiro caso no Brasil, tendo evoluído com epitelização total da córnea e melhora da acuidade visual. Após o 3º mês da cirurgia, iniciou-se neovascularização superficial periférica com piora da transparência corneana. A visão manteve-se 0,1 após um ano de cirurgia.Case report of a patient who developed failure of an allogenic living related conjunctival limbal transplantation in the right eye after an episode of epidemic keratoconjunctivitis. After a few months, she underwent transplantation of allogenic limbal epithelial stem cells cultivated ex vivo on amniotic membrane (first case in Brazil. The patient evolved with total corneal epithelialization and improvement of the visual acuity. Three months after the surgery, peripheral superficial neovascularization with worsening of the corneal transparency was observed. The vision remained 0.1 after one year of the transplantation.

  19. Achieving effective confinement through utilization of non-Newtonian fluid mixture as stemming structure

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    Luís Felipe Gomes Marinho

    Full Text Available Abstract The economics of a mining operation is directly influenced by blasting outcomes, where blasting aims to comminute the rock mass in order to attain smaller grain sizes to be loaded and hauled at a minimum cost for its first processing stage. In order to promote adequate rock breakage, the stemming structure needs to provide proper confinement for the borehole charged with explosives, reflecting the energy released during the detonation in form of shock waves and gases to act throughout the in situ rock mass, enlarging its failures and fractures, and also creating new ones. To build up a stemming column, literature recommends the usage of dry granular materials instead of elements with plastic behavior. However, a study was performed using Gypsum plaster as stemming; a kind of material that exhibits solid-like behavior when it is dry. Following this theory, this test verified improvements regarding confinement effectiveness and energy propagation throughout the rock mass when a non-Newtonian mixture (NNM was applied as stemming; a material that shows a solid-like behavior when is under shear stress. When the stemming arrangement was composed of NNM, it was able to reduce energy and gas losses to the atmosphere, because of the liquid's property of filling voids into the borehole. The NNM yielded high results due to its better confinement effectiveness, a reduction of air overpressure, and an increase of the strain propagation and ground vibration throughout the rock.

  20. Quantitative mapping of intracellular cations in the human amniotic membrane

    Science.gov (United States)

    Moretto, Ph.; Llabador, Y.; Simonoff, M.; Razafindrabe, L.; Bara, M.; Guiet-Bara, A.

    1993-05-01

    The effect of magnesium and taurine on the permeability of cell membranes to monovalent cations has been investigated using the Bordeaux nuclear microprobe. PIXE and RBS techniques have been used to provide quantitative measurements and ion distributions in the isolated amniotic membrane. This physiological model for cellular exchanges allowed us to reveal the distribution of most elements involved in cellular pathways and the modifications under different experimental conditions of incubation in physiological fluids. The PIXE microanalysis provided an original viewpoint on these mechanisms. Following this first study, the amnion compact lamina was found to play a role which was not, up to now, taken into account in the interpretation of electrophysiological experimentations. The release of some ionic species, such as K +, from the epithelial cells, during immersion in isotonic fluids, could have been hitherto underestimated.

  1. Effect of the Human Amniotic Membrane on Liver Regeneration in Rats

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    Mesut Sipahi

    2015-01-01

    Full Text Available Introduction. Operations are performed for broader liver surgery indications for a better understanding of hepatic anatomy/physiology and developments in operation technology. Surgery can cure some patients with liver metastasis of some tumors. Nevertheless, postoperative liver failure is the most feared complication causing mortality in patients who have undergone excision of a large liver mass. The human amniotic membrane has regenerative effects. Thus, we investigated the effects of the human amniotic membrane on regeneration of the resected liver. Methods. Twenty female Wistar albino rats were divided into control and experimental groups and underwent a 70% hepatectomy. The human amniotic membrane was placed over the residual liver in the experimental group. Relative liver weight, histopathological features, and biochemical parameters were assessed on postoperative day 3. Results. Total protein and albumin levels were significantly lower in the experimental group than in the control group. No difference in relative liver weight was observed between the groups. Hepatocyte mitotic count was significantly higher in the experimental group than in the control group. Hepatic steatosis was detected in the experimental group. Conclusion. Applying the amniotic membrane to residual liver adversely affected liver regeneration. However, mesenchymal stem cell research has the potential to accelerate liver regeneration investigations.

  2. Implantation of Neuronal Stem Cells Enhances Object Recognition without Increasing Neurogenesis after Lateral Fluid Percussion Injury in Mice

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    Laura B. Ngwenya

    2018-01-01

    Full Text Available Cognitive deficits after traumatic brain injury (TBI are debilitating and contribute to the morbidity and loss of productivity of over 10 million people worldwide. Cell transplantation has been linked to enhanced cognitive function after experimental traumatic brain injury, yet the mechanism of recovery is poorly understood. Since the hippocampus is a critical structure for learning and memory, supports adult neurogenesis, and is particularly vulnerable after TBI, we hypothesized that stem cell transplantation after TBI enhances cognitive recovery by modulation of endogenous hippocampal neurogenesis. We performed lateral fluid percussion injury (LFPI in adult mice and transplanted embryonic stem cell-derived neural progenitor cells (NPC. Our data confirm an injury-induced cognitive deficit in novel object recognition, a hippocampal-dependent learning task, which is reversed one week after NPC transplantation. While LFPI alone promotes hippocampal neurogenesis, as revealed by doublecortin immunolabeling of immature neurons, subsequent NPC transplantation prevents increased neurogenesis and is not associated with morphological maturation of endogenous injury-induced immature neurons. Thus, NPC transplantation enhances cognitive recovery early after LFPI without a concomitant increase in neuron numbers or maturation.

  3. Tissue Banking in Malaysia-amniotic membrane

    International Nuclear Information System (INIS)

    Hashim bin Mohamad; Norimah binti Yusof

    1991-01-01

    Burn treatment using amniotic membranes in some of our patients initiate our own tissue bank starting with a pilot project on procurement, processing and clinical application of irradiated amniotic membrane. The irradiation of amniotic membrane was made possible by the availability of cobalt source at the Nuclear Energy Agency (UTN). With the technical help from the Inter-national Atomic Energy Agency (IAEA) we soon should be able to embark on bone bank to supply local surgeons. Thus the establishment of tissue bank at our institution will further enhance our programme which will include keratinocytes culture for burn, osteocytes culture for bone replacement as well as the use of animal skin for temporary coverage of open wounds

  4. Comparison study of moisture content, colour properties and essential oil compounds extracted by hydrodistillation and supercritical fluid extraction between stem and leaves of lemongrass (Cymbopogun citratus)

    Science.gov (United States)

    Kamaruddin, Shazlin; Mustapha, Wan Aida Wan; Haiyee, Zaibunnisa Abdul

    2018-04-01

    The objectives of this study were to compare the properties of moisture content, colour and essential oil compounds between stem and leaves of lemongrass (Cymbopogun citratus). The essential oil was extracted using two different methods which are hydrodistillation and supercritical fluid extraction (SFE). There was no significant difference of moisture content between stem and leaves of lemongrass. The lightness (L) and yellowness (+b) values of the stems were significantly higher (pleaves. The highest yield of essential oil was obtained by extraction using supercritical fluid extraction (SFE) in leaves (˜ 0.7%) by treatment at 1700psi and 50°C. The main compound of extracted essential oil was citral (geranial and neral).

  5. Prolonged Harvest Time on Amniotic Cell Culture: Is it Offer Important in Prediction of Fetuses with Trisomies?

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    Rengin Karataylı

    2012-08-01

    CONCLUSION: In this study, initial hypothesis was that there may be a possible association of prolonged harvest time with trisomic chromosomal aberrations. Our results concluded that harvest time does not differ between normal and trisomic amniotic fluid cultures.

  6. Associação entre perfil glicêmico materno e o índice de líquido amniótico em gestações complicadas pelo Diabetes mellituspré-gestacional Association between maternal glycemic profile and amniotic fluid index in pregnancies complicated by pregestational Diabetes Mellitus

    Directory of Open Access Journals (Sweden)

    Carlos Alberto Maganha

    2009-01-01

    Full Text Available OBJETIVOS: Estudar a relação entre o volume de líquido amniótico e o perfil glicêmico em gestantes com Diabetes mellitus tipo 1 e tipo 2 acompanhadas em ambulatório especializado e multidisciplinar. MÉTODOS: Este estudo observacional foi realizado entre janeiro de 2001 e dezembro de 2004. Os critérios de inclusão adotados foram: gestação única, diagnóstico de Diabetes mellitus pré-gestacional, início do pré-natal antes da 26ª semana, ausência de anomalias fetais. Foram excluídos os casos em que o recém-nascido apresentou-se pequeno para a idade gestacional. O índice de líquido amniótico (ILA foi avaliado semanalmente a partir da 27ª semana de gestação até o parto e comparado com o perfil glicêmico da semana precedente ao exame ultrassonográfico. O perfil glicêmico foi analisado pela média glicêmica. A correlação entre o perfil glicêmico e ILA foi analisada pelo índice de Spearman. RESULTADOS: Foram estudadas 60 gestantes, perfazendo um total de 659 correlações entre o ILA e o perfil glicêmico. Em nenhuma idade gestacional estudada houve correlação entre o ILA e o perfil glicêmico. No grupo com ILA 18 cm a média glicêmica foi de 103,67 mg/dl (DP=11,46, não apresentando diferença significativa. CONCLUSÃO: Em gestantes diabéticas tipo 1 e 2, com tratamento padronizado e controle rigoroso metabólico, não houve relação entre o ILA e o perfil glicêmico materno no terceiro trimestre de gestação.BACKGROUND: to study the relation between amniotic fluid volume and glycemic control in pregnancies complicated by diabetes mellitus type 1 and 2, followed in a specialized multidisciplinary prenatal care service. METHODS: This descriptive study was performed between January 2001 and December 2004. Inclusion criteria were: simple pregnancy, diagnosis of pregestational diabetes, beginning of prenatal care before the 26th week and absence of fetal anomaly. Cases with newborns small for gestational age were

  7. The amniotic band syndrome: antenatal sonographic diagnosis and potential pitfalls.

    Science.gov (United States)

    Mahony, B S; Filly, R A; Callen, P W; Golbus, M S

    1985-05-01

    Amniotic band syndrome causes a variety of fetal malformations involving the limbs, craniofacial region, and trunk. Six prenatally diagnosed cases of amniotic band syndrome are discussed. The diagnosis was based on sonographic visualization of either amniotic sheets or bands associated with fetal deformation or deformities in nonembryologic distributions known to characterize the amniotic band syndrome. Seven additional cases are considered in which an aberrant sheet of tissue with a free edge was visualized within the amniotic cavity but no restriction of fetal motion or subsequent deformity was demonstrated.

  8. INTERLEUKIN-6 TRANS-SIGNALING SYSTEM IN INTRA-AMNIOTIC INFLAMMATION, PRETERM BIRTH AND PRETERM PREMATURE RUPTURE OF THE MEMBRANES

    Science.gov (United States)

    Lee, Sarah Y.; Buhimschi, Irina A.; Dulay, Antonette T.; Ali, Unzila A.; Zhao, Guomao; Abdel-Razeq, Sonya S.; Bahtiyar, Mert O.; Thung, Stephen F.; Funai, Edmund F.; Buhimschi, Catalin S.

    2013-01-01

    Classic IL-6 signaling is conditioned by the transmembrane receptor (IL-6R) and homodimerization of gp130. During trans-signaling, IL-6 binds to soluble IL-6R (sIL-6R) enabling activation of cells expressing solely gp130. Soluble gp130 (sgp130) selectively inhibits IL-6 trans-signaling. To characterize amniotic fluid IL-6 trans-signaling molecules (IL-6, sIL-6R, sgp130) in normal gestations and pregnancies complicated by intra-amniotic inflammation (IAI) we studied 301 women during second trimester (n=39), third trimester (n=40) and preterm labor with intact (n=131, 85 IAI negative & 46 IAI positive) or preterm premature rupture of membranes (PPROM: n=91, 61 IAI negative & 30 IAI positive). ELISA, Western blotting and RT-PCR were used to investigate amniotic fluid, placenta and amniochorion for protein and mRNA expression of sIL-6R, sgp130, IL-6R and gp130. Tissues were immunostained for IL-6R, gp130, CD15+ (polymorphonuclear) and CD3+ (T-cell) inflammatory cells. The ability of sIL-6R and sgp130 to modulate basal and LPS-stimulated release of amniochorion matrix-metalloprotease-9 (MMP-9) was tested ex-vivo. We showed that in physiologic gestations amniotic fluid sgp130 decreases toward term. Amniotic fluid IL-6 and sIL-6R were elevated in IAI whereas sgp130 was decreased in PPROM. Our results suggested that fetal membranes are the probable source of amniotic fluid sIL-6R and sgp130. Immunohistochemistry and RT-PCR revealed increased IL-6R and decreased gp130 expression in amniochorion of women with IAI. Ex-vivo, sIL-6R and LPS augmented amniochorion MMP-9 release whereas sgp130 opposed this effect. We conclude that IL-6 trans-signaling molecules are physiologic constituents of the amniotic fluid regulated by gestational age and inflammation. PPROM likely involves functional loss of sgp130. PMID:21282511

  9. Amniotic amputation | Ayadi | Pan African Medical Journal

    African Journals Online (AJOL)

    Amniotic band syndrome (ABS) is an uncommon, congenital fetal abnormality. Lower extremity limb defects are the common manifestations of ABS. The most common features include congenital distal ring constrictions, intrauterine amputations, and acrosyndactyly. Rare cases of craniofacial and visceral defects were ...

  10. Stem cells and fluid flow drive cyst formation in an invertebrate excretory organ.

    Science.gov (United States)

    Thi-Kim Vu, Hanh; Rink, Jochen C; McKinney, Sean A; McClain, Melainia; Lakshmanaperumal, Naharajan; Alexander, Richard; Sánchez Alvarado, Alejandro

    2015-06-09

    Cystic kidney diseases (CKDs) affect millions of people worldwide. The defining pathological features are fluid-filled cysts developing from nephric tubules due to defective flow sensing, cell proliferation and differentiation. The underlying molecular mechanisms, however, remain poorly understood, and the derived excretory systems of established invertebrate models (Caenorhabditis elegans and Drosophila melanogaster) are unsuitable to model CKDs. Systematic structure/function comparisons revealed that the combination of ultrafiltration and flow-associated filtrate modification that is central to CKD etiology is remarkably conserved between the planarian excretory system and the vertebrate nephron. Consistently, both RNA-mediated genetic interference (RNAi) of planarian orthologues of human CKD genes and inhibition of tubule flow led to tubular cystogenesis that share many features with vertebrate CKDs, suggesting deep mechanistic conservation. Our results demonstrate a common evolutionary origin of animal excretory systems and establish planarians as a novel and experimentally accessible invertebrate model for the study of human kidney pathologies.

  11. Surface Characterization and Human Stem Cell Behaviors of Zirconia Implant Disks Biomimetic-Treated in Simulated Body Fluid.

    Science.gov (United States)

    Quan, Hongxuan; Park, Yoon-Kyung; Kim, Seong-Kyun; Heo, Seong-Joo; Koak, Jai-Young; Han, Jung-Suk; Lee, Joo-Hee

    2016-01-01

    This study investigated the effects of biomimetic deposition on a zirconia surface in simulated body fluid (SBF) and assessed the proliferation and differentiation of human bone marrow mesenchymal stem cells on the SBF-treated zirconia disks. Corrected SBF was prepared according to Kokubo's recipe. Eighty yttrium oxide-stabilized tetragonal zirconia polycrystalline disks were prepared and divided into two groups: (1) the test group with SBF-treated disks and (2) the control group with nontreated disks. Zirconia disks were soaked in SBF for 1, 4, 7, and 14 days at 36.5°C, and the hydroxyapatite (HA) precipitation was verified by analyzing the surface morphology. For more in-depth validation of HA formation, the surface roughness, composition, and crystallization of the 7-day treated disks were analyzed. Human bone marrow mesenchymal stem cells were used to further evaluate cell proliferation, alkaline phosphatase activity, and osteoblast gene expression on the 7-day treated zirconia disks. Disks showed different surface morphologies after soaking for different time periods. As the SBF soaking time increased, the amount of HA coverage increased gradually, uniformly covering the disks by day 7. There was no difference in surface roughness between the two groups (P > .05). Cell proliferation was higher on the SBF-treated disks (P .05). This study demonstrated that biomimetic deposition has an effect on the formation of HA on zirconia disks. The cell attachment, proliferation, and differentiation of SBF-treated zirconia disks was superior to that of nontreated disks, which indicates that SBF-treated zirconia implants have long-term clinical value.

  12. Human amniotic epithelial cell transplantation for the repair of injured brachial plexus nerve: evaluation of nerve viscoelastic properties

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    Hua Jin

    2015-01-01

    Full Text Available The transplantation of embryonic stem cells can effectively improve the creeping strength of nerves near an injury site in animals. Amniotic epithelial cells have similar biological properties as embryonic stem cells; therefore, we hypothesized that transplantation of amniotic epithelial cells can repair peripheral nerve injury and recover the creeping strength of the brachial plexus nerve. In the present study, a brachial plexus injury model was established in rabbits using the C 6 root avulsion method. A suspension of human amniotic epithelial cells was repeatedly injected over an area 4.0 mm lateral to the cephal and caudal ends of the C 6 brachial plexus injury site (1 × 10 6 cells/mL, 3 μL/injection, 25 injections immediately after the injury. The results showed that the decrease in stress and increase in strain at 7,200 seconds in the injured rabbit C 6 brachial plexus nerve were mitigated by the cell transplantation, restoring the viscoelastic stress relaxation and creep properties of the brachial plexus nerve. The forepaw functions were also significantly improved at 26 weeks after injury. These data indicate that transplantation of human amniotic epithelial cells can effectively restore the mechanical properties of the brachial plexus nerve after injury in rabbits and that viscoelasticity may be an important index for the evaluation of brachial plexus injury in animals.

  13. Modulation of Synovial Fluid-Derived Mesenchymal Stem Cells by Intra-Articular and Intraosseous Platelet Rich Plasma Administration

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    Emma Muiños-López

    2016-01-01

    Full Text Available The aim of this study was to evaluate the effect of intra-articular (IA or a combination of intra-articular and intraosseous (IO infiltration of Platelet Rich Plasma (PRP on the cellular content of synovial fluid (SF of osteoarthritic patients. Thirty-one patients received a single infiltration of PRP either in the IA space (n=14 or in the IA space together with two IO infiltrations, one in the medial femoral condyle and one in the tibial plateau (n=17. SF was collected before and after one week of the infiltration. The presence in the SF of mesenchymal stem cells (MSCs, monocytes, and lymphocytes was determined and quantified by flow cytometry. The number and identity of the MSCs were further confirmed by colony-forming and differentiation assays. PRP infiltration into the subchondral bone (SB and the IA space induced a reduction in the population of MSCs in the SF. This reduction in MSCs was further confirmed by colony-forming (CFU-F assay. On the contrary, IA infiltration alone did not cause variations in any of the cellular populations by flow cytometry or CFU-F assay. The SF of osteoarthritic patients contains a population of MSCs that can be modulated by PRP infiltration of the SB compartment.

  14. Experience of Using Amniotic Membrane After Circumcision

    International Nuclear Information System (INIS)

    Manjas, Menkher; Ismal; Efmansyah, Dody

    2002-01-01

    It is compulsory, for boys to undergone circumcision before getting adult in Moslem region. It can be done by General Surgeon, General Practitioner, Nurse, Midwife or Quack. The place to carry out the circumcision can be inside or outside hospital. The utmost problems are injections, point for secondary wound covering and delay of using underpants. To overcome those problem amniotic membranes can be used as wound covering, based on : they are soft, easy to shape wound surface, satisfactory adhesive properties, good elasticity and sufficient, transparency which allows wound control without redressing of the wound. From January until December 1999, 165 boys at an age between 6-10 years, which have been carried out circumcision, were evaluated. Radiation sterilized lyophilized amniotic membranes were used in this work as wound covering Result show that amniotic membrane gave a good result in wound healing. All the patients observed, showed early mobilization as well as early using underpants. There is no different result between circumcision which had been done either inside or out hospital, carried out by surgeon or non-surgeon

  15. Comparison of rapid MMP-8 and interleukin-6 point-of-care tests to identify intra-amniotic inflammation/infection and impending preterm delivery in patients with preterm labor and intact membranes.

    Science.gov (United States)

    Chaemsaithong, Piya; Romero, Roberto; Docheva, Nikolina; Chaiyasit, Noppadol; Bhatti, Gaurav; Pacora, Percy; Hassan, Sonia S; Yeo, Lami; Erez, Offer

    2018-01-01

    Among patients presenting with preterm labor and intact membranes, those with intra-amniotic inflammation have adverse obstetrical and neonatal outcomes. The diagnosis of intra-amniotic inflammation can easily be made by detecting an elevated concentration of the cytokine interleukin (IL)-6 or the enzyme neutrophil collagenase, also known as matrix metalloproteinase (MMP)-8. The diagnostic performances of MMP-8 and IL-6 enzyme-linked immunosorbent assay tests are similar. Recently, a rapid test has become available for point-of-care determination of either MMP-8 or IL-6. The objectives of this study were to compare the diagnostic indices and predictive values between the rapid MMP-8 and IL-6 tests for the identification of intra-amniotic inflammation in patients with preterm labor and intact membranes. We performed a retrospective cohort study including 124 women with singleton pregnancies who presented with symptoms of preterm labor and underwent transabdominal amniocentesis for the evaluation of microbial invasion of the amniotic cavity (MIAC). MIAC was defined according to amniotic fluid culture results (aerobic and anaerobic bacteria as well as genital Mycoplasmas). Amniotic fluid white blood cell (WBC) counts were determined using a hemocytometer chamber. An elevated amniotic fluid MMP-8 concentration was assessed using Yoon's MMP-8 Check ® (cutoff: 10 ng/mL). An elevated amniotic fluid IL-6 concentration was scored when there was a positive result for the lateral flow-based immunoassay (cutoff: ≥745 pg/mL and ≥1000 pg/mL). In order to objectively compare rapid MMP-8 and rapid IL-6 tests to identify intra-amniotic inflammation, an amniotic fluid WBC count of ≥50 cells/mm 3 was used to define intra-amniotic inflammation. (1) The rapid tests had the same sensitivity for the detection of intra-amniotic inflammation [85.7% (18/21) for all]; (2) the specificity of the rapid MMP-8 test was higher than that of the rapid IL-6 test (cutoff: 745

  16. Amniotic Membrane Transplantation in Surgical Treatment of Conjunctival Melanoma: Long-term Results.

    Science.gov (United States)

    Palamar, Melis; Yaman, Banu; Akalın, Taner; Yağcı, Ayse

    2018-02-01

    To investigate the long-term efficacy and results of surgical management of conjunctival melanoma reconstructed with amniotic membrane transplantation. Conjunctival melanoma in 10 patients (5 female, 5 male) was totally excised with adjunctive cryotherapy to the surgical margins, corneal epitheliectomy with absolute alcohol in cases of corneal involvement, lamellar sclerectomy in cases with episcleral involvement, and ocular surface grafting with cryopreserved amniotic membrane. Complications and tumor control rates were evaluated. The mean age of the patients was 57.4±15.2 (range, 37-84) years. The mean diameter of the tumors was 15.5±4.9 (range, 10-25) mm and histopathologically confirmed complete excision was performed in all cases. Mild limbal stem cell deficiency (2 eyes) and subclinical symblepharon (3 eyes) were observed as long-term complications. In a mean follow-up of 56.7±40.4 (range, 30-132) months, only one local tumor recurrence was detected. Despite retreatment, exenteration was performed in this patient due to re-recurrence. One patient died due to disseminated metastasis despite the absence of local recurrence. In large conjunctival melanomas, reconstruction of the ocular surface is usually very challenging. The use of cryopreserved amniotic membrane for conjunctival defect repair is safe and effective with mild complications, and allows the excision of wider margins around the tumor.

  17. Amniotic Membrane Transplantation in Surgical Treatment of Conjunctival Melanoma: Long-term Results

    Directory of Open Access Journals (Sweden)

    Melis Palamar

    2018-02-01

    Full Text Available Objectives: To investigate the long-term efficacy and results of surgical management of conjunctival melanoma reconstructed with amniotic membrane transplantation. Materials and Methods: Conjunctival melanoma in 10 patients (5 female, 5 male was totally excised with adjunctive cryotherapy to the surgical margins, corneal epitheliectomy with absolute alcohol in cases of corneal involvement, lamellar sclerectomy in cases with episcleral involvement, and ocular surface grafting with cryopreserved amniotic membrane. Complications and tumor control rates were evaluated. Results: The mean age of the patients was 57.4±15.2 (range, 37-84 years. The mean diameter of the tumors was 15.5±4.9 (range, 10-25 mm and histopathologically confirmed complete excision was performed in all cases. Mild limbal stem cell deficiency (2 eyes and subclinical symblepharon (3 eyes were observed as long-term complications. In a mean follow-up of 56.7±40.4 (range, 30-132 months, only one local tumor recurrence was detected. Despite retreatment, exenteration was performed in this patient due to re-recurrence. One patient died due to disseminated metastasis despite the absence of local recurrence. Conclusion: In large conjunctival melanomas, reconstruction of the ocular surface is usually very challenging. The use of cryopreserved amniotic membrane for conjunctival defect repair is safe and effective with mild complications, and allows the excision of wider margins around the tumor.

  18. Amniotic membrane transplantation in surgical management of ocular surface squamous neoplasias: long-term results.

    Science.gov (United States)

    Palamar, M; Kaya, E; Egrilmez, S; Akalin, T; Yagci, A

    2014-09-01

    To evaluate the long-term efficacy of amniotic membrane transplantation for ocular surface reconstruction in the surgical management of ocular surface squamous neoplasia (OSSN). OSSN in 21 patients (7 female, 14 male patients) was managed with excisional biopsy, cryotherapy, corneal epitheliectomy with absolute alcohol application when the cornea is involved, lamellar sclerectomy and adjunctional absolute alcohol application to the base when episclera is involved, and ocular surface reconstruction with cryopreserved amniotic membrane transplantation. Tumor control and complications were evaluated. The mean age of the patients was 62.42 ± 20.9 (range, 16-84). The average diameter of the base of the tumors was 13.1 ± 4.8 (range, 9-21) mm and complete removal was achieved in all cases as revealed histopathologically. Ocular surface healing was achieved in all cases. At the postoperative period, limbal stem cell deficiency in three eyes and mild symblepharon in one eye were detected. In a mean follow-up of 30.95 ± 18.8 (range, 13-75) months, no recurrence was detected. For large or multifocal conjunctival tumors, the reconstruction of ocular surface and fornix is challenging. The amniotic-membrane use to repair conjunctival defects larger than 10 mm is a safe and effective technique with minimal complications allowing surgeons to make large enough excisions.

  19. Amniotic lamellar body counts can predict the occurrence of respiratory distress syndrome as well as transient tachypnea of the newborn (TTN).

    Science.gov (United States)

    Tsuda, Hiroyuki; Takahashi, Yuichiro; Iwagaki, Shigenori; Uchida, Yasushi; Kawabata, Ichiro; Hayakawa, Masahiro; Sumigama, Seiji; Hayakawa, Hiromi; Kotani, Tomomi; Kikkawa, Fumitaka

    2011-05-01

    The purpose of this study is to predict the occurrence of transient tachypnea of the newborn (TTN) using amniotic lamellar body count (LBC) and compare the LBCs in neonates with TTN with the LBCs in neonates with respiratory distress syndrome (RDS) and controls. Three hundred and eighty-one amniotic fluid samples were obtained at cesarean section from 27 to 40 weeks of gestation. Samples were analyzed immediately without centrifugation and the number of lamellar bodies was counted. The LBC in amniotic fluid ranged from 1,000 to 577,000/μL. An LBC cut-off value of 48,500/μL resulted in 84.7% sensitivity, 76.2% specificity, and 98.1% negative predictive value for predicting TTN. The LBC in neonates with TTN was significantly lower than that in controls (50,000 vs. 122,000; Prespiratory management is required.

  20. Ultrasound estimation of amniotic fluid and perinatal outcome in ...

    African Journals Online (AJOL)

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  1. Elemental Profile in Amniotic Fluid of some Nigerian Pregnant Women

    African Journals Online (AJOL)

    The efficiency of extraction of trace metals using conventional wet acid digestion method (CDM) and microwave induced acid digestion method (MWD) was determined by recovery experiments. Levels of trace metals were determined using Atomic Absorption Spectrophotometry. The high percentage recoveries obtained ...

  2. Estimating Lecithin/Sphingomyelin Area Ratio in Amniotic Fluid

    African Journals Online (AJOL)

    1973-09-24

    Sep 24, 1973 ... fusion which carries a 6,4% risk" of traumatic fetal death. However, if a low LSAR is obtained, results would· favour a fetal blood transfusion, thus allowing ... and examined for bubbles after 15 minutes. The obvious advantages of this method are that it is extremely quick and can be carried out by unskilled ...

  3. the cord blood amniotic fluid the Complement activity of term ...

    African Journals Online (AJOL)

    nic intervillositis), fetal (chorionic and/or umbilical vasculitis) or mixed. A more detailed description of the method ofgrading has been published elsewhere.2. The placentas of the 11 infants with the AFIS had grade 2-3 inflammatory changes, whereas those of the II infants in the control group showed neither maternal nor ...

  4. Mouse embryos cultured in amniotic fluid | Oettle | South African ...

    African Journals Online (AJOL)

    The embryos were cultured for 72 hours at 37°C in 5% carbon dioxide in air. Osmolarity, pH, partial arterial carbon dioxide pressure and HCO; were checked before culture and again at the end of culture. Embryos were assessed according to the stage of development, namely degenerate embryos, morulae and blastocysts.

  5. Estimating Lecithin/Sphingomyelin Area Ratio in Amniotic Fluid ...

    African Journals Online (AJOL)

    A knowledge of the pulmonary maturity of the unborn fetus is an important factor in deciding when to induce birth. A lecithin/sphingomyelin area ratio is obtained by thin-layer chromatography. A molybdenum spray is used to make visible only the choline-containing phospholipids, without necessitating the use of heat. This is ...

  6. Mouse embryos cultured In amnIotIc fluid

    African Journals Online (AJOL)

    Mouse embryos are used as a monitor of human in vicro fertilisation (IVF) culture conditions. Embryo culture media are often complicated solutions requiring careful monitoring to ensure uniformity for successful embryo culture.1-3 Much of the quality control is outside the scope of the IVF laboratory, for example the source of ...

  7. Meconium-stained amniotic fluid – what is the evidence ...

    African Journals Online (AJOL)

    Opinions regarding the significance of meconium-stained liquor detected during labour have varied although there is consensus that meconium aspirated into the lungs of the neonate may lead to meconium aspiration syndrome. The efficacy of various interventions designed to prevent meconium aspiration syndrome are ...

  8. Management strategy in case of meconium stained amniotic fluid (abstract)

    NARCIS (Netherlands)

    Pop, V.J.M.; Kuppens, S.M.

    2014-01-01

    The management strategies can generally be divided in two categories: preventive intervention and “wait and watch” strategy. The first group again can be divided in two categories: the prevention of MSAF to occur and the prevention of any fetal or neonatal complication once MSAF is diagnosed. The

  9. Major complication after intrauterine vesico-amniotic shunting ...

    African Journals Online (AJOL)

    Major complication after intrauterine vesico-amniotic shunting. A Springer, R Fartacek, CA Reck, E Horcher, D Bettelheim. Abstract. Bilateral foetal uropathy is the leading cause of chronic renal failure in childhood. Vesico-amniotic shunting (VAS) is a simple, feasible, and widely used procedure for decompressing the foetal ...

  10. Peculiarity of Porcine Amniotic Membrane and Its Derived Cells: A Contribution to the Study of Cell Therapy from a Large Animal Model.

    Science.gov (United States)

    Lange-Consiglio, Anna; Corradetti, Bruna; Bertani, Sabrina; Notarstefano, Valentina; Perrini, Claudia; Marini, Maria Giovanna; Arrighi, Silvana; Bosi, Giampaolo; Belloli, Angelo; Pravettoni, Davide; Locatelli, Valentina; Cremonesi, Fausto; Bizzaro, Davide

    2015-12-01

    The aim of this work was to provide, for the first time, a protocol for isolation and characterization of stem cells from porcine amniotic membrane in view of their potential uses in regenerative medicine. From three samples of allanto-amnion recovered at delivery, the amniotic membrane was stripped from overlying allantois and digested with trypsin and collagenase to isolate epithelial (amniotic epithelial cells [AECs]) and mesenchymal cells, respectively. Proliferation, differentiation, and characterization studies by molecular biology and flow cytometry were performed. Histological examination revealed very few mesenchymal cells in the stromal layer, and a cellular yield of AECs of 10 × 10(6)/gram of digested tissue was achieved. AECs readily attached to plastic culture dishes displaying typical cuboidal morphology and, although their proliferative capacity decreased to the fifth passage, AECs showed a mean doubling time of 24.77 ± 6 h and a mean frequency of one fibroblast colony-forming unit (CFU-F) for every 116.75 plated cells. AECs expressed mesenchymal stem cell (MSC) mRNA markers (CD29, CD166, CD90, CD73, CD117) and pluripotent markers (Nanog and Oct 4), whereas they were negative for CD34 and MHCII. Mesodermic, ectodermic, and endodermic differentiation was confirmed by staining and expression of specific markers. We conclude that porcine amniotic membrane can provide an attractive source of stem cells that may be a useful tool for biomedical research.

  11. Application of human amniotic mesenchymal cells as an allogeneic transplantation cell source in bone regenerative therapy

    Energy Technology Data Exchange (ETDEWEB)

    Tsuno, Hiroaki [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Department of Oral and Maxillofacial Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Yoshida, Toshiko [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Nogami, Makiko [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Department of Orthopedic Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Koike, Chika; Okabe, Motonori [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Noto, Zenko [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Department of Oral and Maxillofacial Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Arai, Naoya; Noguchi, Makoto [Department of Oral and Maxillofacial Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Nikaido, Toshio, E-mail: tnikaido@med.u-toyama.ac.jp [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan)

    2012-12-01

    Autogenous mesenchymal stem cells (MSCs) have therapeutic applications in bone regenerative therapy due to their pluripotency. However, the ability of MSCs to proliferate and differentiate varies between donors. Furthermore, alternative sources of MSCs are required for patients with contraindications to autogenous cell therapy. The aim of this study was to evaluate the potential of mesenchymal cells from the human amniotic membrane (HAM) as a source of cells for allogeneic transplantation in bone regenerative therapy. Cells that retained a proliferative capacity of more than 50 population doubling level were distinguished from other HAM cells as HAM{alpha} cells and induced to osteogenic status-their in vivo osteogenesis was subsequently investigated in rats. It was found that HAM{alpha} cells were spindle shaped and were positive for MSC markers and negative for hematopoietic stem cell markers. Alkaline phosphatase activity and calcium deposition increased with osteogenic status of HAM{alpha} cells. The expression of osteocalcin mRNA was increased in HAM{alpha} cells cultured on calcium phosphate scaffolds. Moreover, xenografted HAM{alpha} cells remained viable and produced extracellular matrix for several weeks. Thus, this study suggests that human amniotic mesenchymal cells possess osteogenic differentiation potential and could be applied to allogeneic transplantation in bone regenerative therapy. - Highlights: Black-Right-Pointing-Pointer Human amniotic mesenchymal cells include cells (HAM{alpha} cells) that have the properties of MSCs. Black-Right-Pointing-Pointer HAM{alpha} cells have excellent osteogenic differentiation potential. Black-Right-Pointing-Pointer Osteogenic differentiation ability of HAM{alpha} was amplified by calcium phosphate scaffolds. Black-Right-Pointing-Pointer HAM{alpha} cells can be applicable to allogeneic cell transplantation in bone regenerative therapy.

  12. A "sweet-spot" for fluid-induced oscillations in the conditioning of stem cell-based engineered heart valve tissues.

    Science.gov (United States)

    Williams, Alexander; Nasim, Sana; Salinas, Manuel; Moshkforoush, Arash; Tsoukias, Nikolaos; Ramaswamy, Sharan

    2017-12-08

    Fluid-induced shear stresses are involved in the development of cardiovascular tissues. In a tissue engineering framework, this stimulus has also been considered as a mechanical regulator of stem cell differentiation. We recently demonstrated that the fluid-oscillating effect in combination with a physiologically-relevant shear stress magnitude contributes to the formation of stem cell-derived de novo heart valve tissues. However, the range of oscillations necessary to induce favorable gene expression and engineered tissue formation is unknown. In this study, we took a computational approach to establish a range of oscillatory shear stresses that may optimize in vitro valvular tissue growth. Taking a biomimetic approach, three physiologically-relevant flow waveforms from the human: (i) aorta, (ii) pulmonary artery and (iii) superior vena cava were utilized to simulate pulsatile flow conditions within a bioreactor that housed 3 tissue specimens. Results were compared to non-physiological pulsatile flow (NPPF) and cyclic flexure-steady flow (Flex-Flow) conditions. The oscillatory shear index (OSI) was used to quantify the fluid-induced oscillations occurring on the specimen surfaces. The range of mean OSI under the physiological conditions investigated was found to be 0.18 ≤ OSI ≤ 0.23. On the other hand, NPPF and Flex-Flow environments yielded a mean OSI of 0.37 and 0.11 respectively, which were 46% higher and 45% lower than physiological conditions. Moreover, we subsequently conducted OSI-based human bone marrow stem cell (HBMSC) culture experiments which resulted in preferential valvular gene expression and phenotype (significant upregulation of BMP, KLF2A, CD31 and α-SMA using an OSI of 0.23 in comparison to a lower OSI of 0.10 or a higher OSI of 0.38; p OSI exists in the mechanical conditioning of tissue engineered heart valves grown from stem cell sources. We conclude that in vitro heart valve matrix development could be further enhanced by simultaneous

  13. The evolution of tail weaponization in amniotes.

    Science.gov (United States)

    Arbour, Victoria M; Zanno, Lindsay E

    2018-01-31

    Weaponry, for the purpose of intraspecific combat or predator defence, is one of the most widespread animal adaptations, yet the selective pressures and constraints governing its phenotypic diversity and skeletal regionalization are not well understood. Here, we investigate the evolution of tail weaponry in amniotes, a rare form of weaponry that nonetheless evolved independently among a broad spectrum of life including mammals, turtles and dinosaurs. Using phylogenetic comparative methods, we test for links between morphology, ecology and behaviour in extant amniotes known to use the tail as a weapon, and in extinct taxa bearing osseous tail armaments. We find robust ecological and morphological correlates of both tail lashing behaviour and bony tail weaponry, including large body size, body armour and herbivory, suggesting these life-history parameters factor into the evolution of antipredator behaviours and tail armaments. We suggest that the evolution of tail weaponry is rare because large, armoured herbivores are uncommon in extant terrestrial faunas, as they have been throughout evolutionary history. © 2018 The Author(s).

  14. Osteogenic differentiation of amniotic epithelial cells: synergism of pulsed electromagnetic field and biochemical stimuli.

    Science.gov (United States)

    Wang, Qian; Wu, Wenchao; Han, Xiaoyu; Zheng, Ai; Lei, Song; Wu, Jiang; Chen, Huaiqing; He, Chengqi; Luo, Fengming; Liu, Xiaojing

    2014-08-11

    Pulsed electromagnetic field (PEMF) is a non-invasive physical therapy used in the treatment of fracture nonunion or delayed healing. PEMF can facilitate the osteogenic differentiation of bone marrow mesenchymal stem cells in vitro. Amniotic epithelial cells (AECs) have been proposed as a potential source of stem cells for cell therapy. However, whether PEMF could modulate the osteogenic differentiation of AECs is unknown. In the present study, the effects of PEMF on the osteogenic differentiation of AECs were investigated. AECs were isolated from amniotic membrane of human placenta by trypsin digestion and were induced by PEMF and/or osteo-induction medium. After 21 days we used real time RT-PCR and immunocytochemistry to study the expression of osteoblast markers. The signal transduction of osteogenesis was further investigated. The PEMF stimulation, or osteo-induction medium alone could induce osteogenic differentiation of AECs, as shown by expression of osteoblast specific genes and proteins including alkaline phosphatase and osteocalcin. Furthermore, a combination of PEMF and osteo-induction medium had synergy effects on osteogenic differentiation. In our study, the gene expression of BMP-2, Runx2, β-catenin, Nrf2, Keap1 and integrinβ1 were up-regulated in the osteogenic differentiation of AECs induced by PEMF and/or osteo-induction medium. Combined application of PEMF and osteo-induction medium is synergistic for the osteogenic differentiation of AECs. It might be a novel approach in the bone regenerative medicine.

  15. Mapping the mechanome of live stem cells using a novel method to measure local strain fields in situ at the fluid-cell interface.

    Directory of Open Access Journals (Sweden)

    Min Jae Song

    Full Text Available During mesenchymal condensation, the initial step of skeletogenesis, transduction of minute mechanical forces to the nucleus is associated with up or down-regulation of genes, ultimately resulting in formation of the skeletal template and appropriate cell lineage commitment. The summation of these biophysical cues affects the cell's shape and fate. Here, we predict and measure surface strain, in live stem cells, in response to controlled delivery of stresses, providing a platform to direct short-term structure--function relationships and long-term fate decisions. We measure local strains on stem cell surfaces using fluorescent microbeads coated with Concanavalin A. During delivery of controlled mechanical stresses, 4-Dimensional (x,y,z,t displacements of the bound beads are measured as surface strains using confocal microscopy and image reconstruction. Similarly, micro-particle image velocimetry (μ-piv is used to track flow fields with fluorescent microspheres. The measured flow velocity gradient is used to calculate stress imparted by fluid drag at the surface of the cell. We compare strain measured on cell surfaces with those predicted computationally using parametric estimates of the cell's elastic and shear modulus. Finally, cross-correlating stress--strain data to measures of gene transcription marking lineage commitment enables us to create stress--strain--fate maps, for live stem cells in situ. The studies show significant correlations between live stem cell stress--strain relationships and lineage commitment. The method presented here provides a novel means to probe the live stem cell's mechanome, enabling mechanistic studies of the role of mechanics in lineage commitment as it unfolds.

  16. Choriodecidual group B streptococcal inoculation induces fetal lung injury without intra-amniotic infection and preterm labor in Macaca nemestrina.

    Directory of Open Access Journals (Sweden)

    Kristina M Adams Waldorf

    Full Text Available BACKGROUND: Early events leading to intrauterine infection and fetal lung injury remain poorly defined, but may hold the key to preventing neonatal and adult chronic lung disease. Our objective was to establish a nonhuman primate model of an early stage of chorioamnionitis in order to determine the time course and mechanisms of fetal lung injury in utero. METHODOLOGY/PRINCIPAL FINDINGS: Ten chronically catheterized pregnant monkeys (Macaca nemestrina at 118-125 days gestation (term=172 days received one of two treatments: 1 choriodecidual and intra-amniotic saline (n=5, or 2 choriodecidual inoculation of Group B Streptococcus (GBS 1×10(6 colony forming units (n=5. Cesarean section was performed regardless of labor 4 days after GBS or 7 days after saline infusion to collect fetal and placental tissues. Only two GBS animals developed early labor with no cervical change in the remaining animals. Despite uterine quiescence in most cases, blinded review found histopathological evidence of fetal lung injury in four GBS animals characterized by intra-alveolar neutrophils and interstitial thickening, which was absent in controls. Significant elevations of cytokines in amniotic fluid (TNF-α, IL-8, IL-1β, IL-6 and fetal plasma (IL-8 were detected in GBS animals and correlated with lung injury (p<0.05. Lung injury was not directly caused by GBS, because GBS was undetectable in amniotic fluid (~10 samples tested/animal, maternal and fetal blood by culture and polymerase chain reaction. In only two cases was GBS cultured from the inoculation site in low numbers. Chorioamnionitis occurred in two GBS animals with lung injury, but two others with lung injury had normal placental histology. CONCLUSIONS/SIGNIFICANCE: A transient choriodecidual infection can induce cytokine production, which is associated with fetal lung injury without overt infection of amniotic fluid, chorioamnionitis or preterm labor. Fetal lung injury may, thus, occur silently without

  17. Effects of perivitelline fluid obtained from Horseshoe crab on the proliferation and genotoxicity of dental pulp stem cells

    Digital Repository Service at National Institute of Oceanography (India)

    Musa, M.; Ali, K.M.; Kannan, T.P.; Azlina, A.; Omar, N.S.; Chatterji, A.; Mokhtar, K.I.

    Perivitelline fluid (PVF) of the horseshoe crab embryo has been reported to possess an important role during embryogenesis by promoting cell proliferation. This study aims to evaluate the effect of PVF on the proliferation, chromosome aberration (CA...

  18. Fetal stem cells and skeletal muscle regeneration: a therapeutic approach

    Directory of Open Access Journals (Sweden)

    Michela ePozzobon

    2014-08-01

    Full Text Available More than 40% of the body mass is represented by muscle tissue, which possesses the innate ability to regenerate after damage through the activation of muscle specific stem cell, namely satellite cells. Muscle diseases, in particular chronic degenerative state of skeletal muscle such as dystrophies, lead to a perturbation of the regenerative process, which causes the premature exhaustion of satellite cell reservoir due to continue cycles of degeneration/regeneration. Nowadays, the research is focused on different therapeutic approaches, ranging from gene and cell to pharmacological therapy, but still there is not a definitive cure in particular for genetic muscle disease. Taking this in mind, in this article we will give special consideration to muscle diseases and the use of fetal derived stem cells as new approach for therapy. Cells of fetal origin, from cord blood to placenta and amniotic fluid, can be easily obtained without ethical concern, expanded and differentiated in culture, and possess immunemodulatory properties. The in vivo approach in animal models can be helpful to study the mechanism underneath the operating principle of the stem cell reservoir, namely the niche, which holds great potential to understand the onset of muscle pathologies.

  19. Oscillatory fluid flow induces the osteogenic lineage commitment of mesenchymal stem cells: The effect of shear stress magnitude, frequency, and duration.

    Science.gov (United States)

    Stavenschi, Elena; Labour, Marie-Noelle; Hoey, David A

    2017-04-11

    A potent regulator of bone anabolism is physical loading. However, it is currently unclear whether physical stimuli such as fluid shear within the marrow cavity is sufficient to directly drive the osteogenic lineage commitment of resident mesenchymal stem cells (MSC). Therefore, the objective of the study is to employ a systematic analysis of oscillatory fluid flow (OFF) parameters predicted to occur in vivo on early MSC osteogenic responses and late stage lineage commitment. MSCs were exposed to OFF of 1Pa, 2Pa and 5Pa magnitudes at frequencies of 0.5Hz, 1Hz and 2Hz for 1h, 2h and 4h of stimulation. Our findings demonstrate that OFF elicits a positive osteogenic response in MSCs in a shear stress magnitude, frequency, and duration dependent manner that is gene specific. Based on the mRNA expression of osteogenic markers Cox2, Runx2 and Opn after short-term fluid flow stimulation, we identified that a regime of 2Pa shear magnitude and 2Hz frequency induces the most robust and reliable upregulation in osteogenic gene expression. Furthermore, long-term mechanical stimulation utilising this regime, elicits a significant increase in collagen and mineral deposition when compared to static control demonstrating that mechanical stimuli predicted within the marrow is sufficient to directly drive osteogenesis. Copyright © 2017. Published by Elsevier Ltd.

  20. Clinical chorioamnionitis at term VI: acute chorioamnionitis and funisitis according to the presence or absence of microorganisms and inflammation in the amniotic cavity

    Science.gov (United States)

    Romero, Roberto; Chaemsaithong, Piya; Docheva, Nikolina; Korzeniewski, Steven J.; Kusanovic, Juan P.; Yoon, Bo Hyun; Kim, Jung-Sun; Chaiyasit, Noppadol; Ahmed, Ahmed I.; Qureshi, Faisal; Jacques, Suzanne M.; Kim, Chong Jai; Hassan, Sonia S.; Chaiworapongsa, Tinnakorn; Yeo, Lami; Kim, Yeon Mee

    2017-01-01

    Objective Neonates born to mothers with clinical chorioamnionitis at term are at an increased risk of infection. Acute subchorionitis, chorioamnionitis, and funisitis are considered placental histologic features consistent with acute inflammation according to the Society for Pediatric Pathology. The objectives of this study were to examine the performance of placental histologic features in the identification of: 1) microbial-associated intra-amniotic inflammation (intra-amniotic infection); and 2) a fetal inflammatory response syndrome (FIRS). Methods and Materials This retrospective cohort study included women with the diagnosis of clinical chorioamnionitis at term (n=45), who underwent an amniocentesis to determine: 1) the presence of microorganisms using both cultivation and molecular biologic techniques [polymerase chain reaction (PCR) with broad range primers]; and 2) interleukin (IL)-6 concentrations by enzyme-linked immunosorbent assay (ELISA). The diagnostic performance (sensitivity, specificity, accuracy, and likelihood ratios) of placental histologic features consistent with acute inflammation was determined for the identification of microbial-associated intra-amniotic inflammation and FIRS. Results 1) The presence of acute histologic chorioamnionitis and funisitis was associated with the presence of proven intra-amniotic infection assessed by amniotic fluid analysis; 2) funisitis was also associated with the presence of FIRS; 3) the negative predictive value of acute funisitis ≥ stage 2 for the identification of neonates born to mothers with intra-amniotic infection was less than 50%, and therefore, suboptimal to exclude fetal exposure to bacteria in the amniotic cavity; and 4) acute funisitis ≥ stage 2 had a negative predictive value of 86.8% for the identification of FIRS in a population with a prevalence of 20%. Conclusion Acute histologic chorioamnionitis and funisitis are associated with intra-amniotic infection and the presence of FIRS

  1. Indications and Outcomes of Amniotic Membrane Transplantation

    Directory of Open Access Journals (Sweden)

    Alime Güneş

    2014-03-01

    Full Text Available Objectives: To investigate the indications and outcomes of amniotic membrane transplantation (AMT. Materials and Methods: A total of 87 eyes who underwent amnion membrane transplantation in our clinic between February 2010 and April 2013 were included in this study. Mean follow-up period was 7.49±7.84 months (range 1-29 months. Amnion membrane defrosted at room temperature and washed three times with saline covered all over the cornea independent of the position and size of the lesion and was sutured to the peripheral cornea circumferentially by 10/0 monofilament suture. At the end of the operation, therapeutic contact lens was placed. All cases were evaluated with respect to duration of surface epithelial healing, visual acuity, presence of recurrence, and infection. Results: Forty-six of the 87 patients (52.8% were men and 41 (47.1% were women. The mean age was 57.02±19.74 years (range 31-89 years in men and 66.69±16.56 years (range 8-88 years in women. The indications were: ulcers in 27 eyes, keratitis in 24 eyes, topical anesthetic abuse in 10 eyes, bullous keratopathy in 7 eyes, conjunctival mass in 7 eyes, symblepharon in 3 eyes, chemical burns in 3 eyes, pterygium in 2 eyes, endophthalmitis in 2 eyes, dellen in 1 eye, and conjunctival laceration in 1 eye. After AMT, 12 patients required second AMT, and 2 cases required third AMT. 2 eyes were eviscerated, 1 eye was exenterated. At the final follow-up visit, except for patients who underwent evisceration and exenteration, improved visual acuity was observed in 45 of the 84 eyes (53.5%. The average of healing time was between 4 and 6 weeks. No infectious, inflammatory, immunologic, or toxic/allergic reactions related to AMT was observed. Conclusion: Amniotic membrane transplantation is a safe and effective technique in ocular surface diseases. (Turk J Ophthalmol 2014; 44: 123-6

  2. Allogeneic amniotic membrane-derived mesenchymal stromal cell transplantation in a porcine model of chronic myocardial ischemia

    Directory of Open Access Journals (Sweden)

    Kimura M

    2012-01-01

    Full Text Available Introduction. Amniotic membrane contains a multipotential stem cell population and is expected to possess the machinery to regulate immunological reactions. We investigated the safety and efficacy of allogeneic amniotic membrane-derived mesenchymal stromal cell (AMSC transplantation in a porcine model of chronic myocardial ischemia as a preclinical trial. Methods. Porcine AMSCs were isolated from amniotic membranes obtained by cesarean section just before delivery and were cultured to increase their numbers before transplantation. Chronic myocardial ischemia was induced by implantation of an ameroid constrictor around the left circumflex coronary artery. Four weeks after ischemia induction, nine swine were assigned to undergo either allogeneic AMSC transplantation or normal saline injection. Functional analysis was performed by echocardiography, and histological examinations were carried out by immunohistochemistry 4 weeks after AMSC transplantation. Results. Echocardiography demonstrated that left ventricular ejection fraction was significantly improved and left ventricular dilatation was well attenuated 4 weeks after AMSC transplantation. Histological assessment showed a significant reduction in percentage of fibrosis in the AMSC transplantation group. Injected allogeneic green fluorescent protein (GFP-expressing AMSCs were identified in the immunocompetent host heart without the use of any immunosuppressants 4 weeks after transplantation. Immunohistochemistry revealed that GFP colocalized with cardiac troponin T and cardiac troponin I. Conclusions. We have demonstrated that allogeneic AMSC transplantation produced histological and functional improvement in the impaired myocardium in a porcine model of chronic myocardial ischemia. The transplanted allogeneic AMSCs survived without the use of any immunosuppressants and gained cardiac phenotype through either their transdifferentiation or cell fusion.

  3. Amniotic membrane in oral and maxillofacial surgery.

    Science.gov (United States)

    Kesting, Marco Rainer; Wolff, Klaus-Dietrich; Nobis, Christopher Philipp; Rohleder, Nils Hagen

    2014-06-01

    Following its renaissance in ophthalmology during the 1990s, preserved human amniotic membrane (HAM) has become an attractive biomaterial for all surgical disciplines. This article reviews the current and potential use of HAM in oral and maxillofacial surgery, its postulated properties and common preservation techniques. Literature was identified by an electronic search of PubMed in July 2012; this was supplemented from the reference lists of the consulted papers. HAM has been used in the field of oral and maxillofacial surgery from 1969 onwards because of its immunological preference and its pain-reducing, antimicrobial, mechanical and side-dependent adhesive or anti-adhesive properties. The effects of HAM on dermal and mucosal re-epithelialisation have been highlighted. Typically, HAM is applied after being banked in a glycerol-preserved, DMSO-preserved or freeze-dried and irradiated state. Whereas the use of HAM in flap surgery and in intra-oral and extra-oral lining is reported frequently, novel HAM applications in post-traumatic orbital surgery and temporomandibular joint surgery have been added since 2010. Tissue engineering with HAM is a fast-expanding field with a high variety of future options. Preserved HAM is considered to be a safe and sufficient biomaterial in all fields of oral and maxillofacial wound healing. Recently published novel indications for HAM application lack a high level of evidence and need to be studied further.

  4. Surface functionalization of tissue culture polystyrene plates with hydroxyapatite under body fluid conditions and its effect on differentiation behaviors of mesenchymal stem cells.

    Science.gov (United States)

    Iijima, Kazutoshi; Suzuki, Ryo; Iizuka, Ayako; Ueno-Yokohata, Hitomi; Kiyokawa, Nobutaka; Hashizume, Mineo

    2016-11-01

    The surfaces of polystyrene (PS) cell culture plates were functionalized with hydroxyapatite (HAp) under body fluid conditions utilizing protein adsorption layers and a pretreatment with an alternate soaking process (ASP) using solutions containing calcium and phosphate ions. Adsorption layers of human serum albumin (HSA) formed on the surface of each well of commercial 24-well PS plates by solution processes. CaCl2 and K2HPO4 solutions were alternately added to the wells, the plates were incubated to form the precursors, and this was followed by the addition of simulated body fluid (SBF) and a further incubation for 24h. These treatments resulted in the surfaces of the PS cell culture plates being completely covered with bone-like HAp. The coating of PS plates with HAp promoted the adhesion of mesenchymal stem cells (MSCs) and maintained cell growth that was as fast as that on tissue culture-treated PS (TCPS) plates. Osteogenic differentiation was greater, whereas adipogenic and chondrogenic differentiation was less in the culture on HAp-coated PS plates than in that on TCPS plates. The present method is useful for preparing HAp-coated PS plates at clean benches without the need for any expensive apparatus. HAp coated on PS plates by this method was a bone-like apatite with high bioactivity; therefore, the present HAp-coated PS plates are promising materials for assays of bone-related cells in the bone remodeling process. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Cord blood stem cell banking: a snapshot of the Italian situation.

    Science.gov (United States)

    Capone, Francesca; Lombardini, Letizia; Pupella, Simonetta; Grazzini, Giuliano; Costa, Alessandro Nanni; Migliaccio, Giovanni

    2011-09-01

    In Italy, the law does not permit the setting up of private banks to preserve cord blood (CB) stem cells for personal use. However, since 2007 the right to export and preserve them in private laboratories located outside Italy has existed, and an increasing number of women are requesting this collection of umbilical CB at delivery to enable storage of stem cells for autologous use. Since private banks recruit clients mainly via the Internet, we examined the content of 24 Italian-language websites that offer stem cells storage (from CB or amniotic fluid), to assess what information is available. We found that the majority of private banks give no clear information about the procedures of collection, processing, and banking of CB units and that the standards offered by private CB banks strongly differ in terms of exclusion or acceptance criteria from the public banks. These factors may well influence the overall quality of the CB units stored in private CB banks. Of note, during the period 2007 to 2009, the number collected for autologous use did not create a downward trend on the number of units stored in public CB banks for allogeneic use. CB is a valuable community resource but expectant parents should be better informed as to the quality variables necessary for its storage, both by institutions and by professionals. Currently, most of the advertising is insufficient to justify the expense and the hopes pinned on autologous use of CB stem cells. © 2011 American Association of Blood Banks.

  6. 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

  7. GROUP B STREPTOCOCCUS CIRCUMVENTS NEUTROPHILS AND NEUTROPHIL EXTRACELLULAR TRAPS DURING AMNIOTIC CAVITY INVASION AND PRETERM LABOR

    Science.gov (United States)

    Boldenow, Erica; Gendrin, Claire; Ngo, Lisa; Bierle, Craig; Vornhagen, Jay; Coleman, Michelle; Merillat, Sean; Armistead, Blair; Whidbey, Christopher; Alishetti, Varchita; Santana-Ufret, Veronica; Ogle, Jason; Gough, Michael; Srinouanprachanh, Sengkeo; MacDonald, James W; Bammler, Theo K; Bansal, Aasthaa; Liggitt, H. Denny; Rajagopal, Lakshmi; Waldorf, Kristina M Adams

    2016-01-01

    Preterm birth is a leading cause of neonatal morbidity and mortality. Although microbial invasion of the amniotic cavity (MIAC) is associated with the majority of early preterm births, the temporal events that occur during MIAC and preterm labor are not known. Group B Streptococci (GBS) are β-hemolytic, gram-positive bacteria, which commonly colonize the vagina but have been recovered from the amniotic fluid in preterm birth cases. To understand temporal events that occur during MIAC, we utilized a unique chronically catheterized nonhuman primate model that closely emulates human pregnancy. This model allows monitoring of uterine contractions, timing of MIAC and immune responses during pregnancy-associated infections. Here, we show that adverse outcomes such as preterm labor, MIAC, and fetal sepsis were observed more frequently during infection with hemolytic GBS when compared to nonhemolytic GBS. Although MIAC was associated with systematic progression in chorioamnionitis beginning with chorionic vasculitis and progressing to neutrophilic infiltration, the ability of the GBS hemolytic pigment toxin to induce neutrophil cell death and subvert killing by neutrophil extracellular traps (NETs) in placental membranes in vivo facilitated MIAC and fetal injury. Furthermore, compared to maternal neutrophils, fetal neutrophils exhibit decreased neutrophil elastase activity and impaired phagocytic functions to GBS. Collectively, our studies demonstrate how a unique bacterial hemolytic lipid toxin enables GBS to circumvent neutrophils and NETs in placental membranes to induce fetal injury and preterm labor. PMID:27819066

  8. Noninvasive prediction of intra-amniotic infection and/or inflammation in preterm premature rupture of membranes.

    Science.gov (United States)

    Park, Kyo Hoon; Kim, Shi Nae; Oh, Kyung Joon; Lee, Sung Youn; Jeong, Eun Ha; Ryu, Aeli

    2012-06-01

    To develop a model based on noninvasive parameters to predict the probability of intra-amniotic infection and/or inflammation (IAI) in women with preterm premature rupture of membranes (PPROMs). Maternal blood was collected for determination of the C-reactive protein (CRP) level and white blood cell (WBC) count immediately after amniocentesis in 171 consecutive women with PPROMs. Intra-amniotic infection and/or inflammation was defined as a positive amniotic fluid (AF) culture and/or an elevated AF interleukin 6 level (≥2.6 ng/mL). A risk score based on a model including maternal blood CRP, WBC, parity, and gestational age was calculated for each patient. The model was shown to have an adequate goodness of fit (P = .516), and the area under the receiver-operating characteristic curve was 0.848, indicating very good discrimination. The noninvasive model based on maternal blood CRP, WBC, parity, and gestational age is highly predictive of IAI in women with PPROMs.

  9. The amniote temporal roof and the diapsid origin of the turtle skull.

    Science.gov (United States)

    Bever, G S; Lyson, Tyler R; Field, Daniel J; Bhullar, Bhart-Anjan S

    2016-12-01

    Fossils provide a glimpse into the architecturally complex origins of modern vertebrate body plans. One such origin that has been long debated is that of turtles. Although much attention has been directed toward the origin of the shell, the enigmatic evolution of the turtle skull and its anapsid temporal region has long clouded our understanding of reptile phylogeny. Two taxa, Eunotosaurus africanus and Pappochelys rosinae, were recently and independently described as long-anticipated stem turtles whose diapsid skulls would cement the evolutionary link between turtles and other modern reptile lineages. Detailed μCT analysis of the stratigraphically older and phylogenetically stemward of the two, Eunotosaurus, provides empirical insight into changing developmental trajectories that may have produced the anapsid cranial form of modern turtles and sets the stage for more comprehensive studies of early amniote cranial evolution. Copyright © 2016 Elsevier GmbH. All rights reserved.

  10. A rare combination of amniotic constriction band with osteogenesis imperfecta.

    Science.gov (United States)

    Shah, Krupa Hitesh; Shah, Hitesh

    2015-11-11

    Amniotic constriction bands and osteogenesis imperfecta are disorders arising from a collagen defect. We report a rare association of amniotic bands with osteogenesis imperfecta in a child. The child was born with multiple amniotic bands involving the right leg, both hands and both feet. Multiple fractures of long bones of lower limbs occurred in childhood due to trivial trauma. Deformities of the femur and tibia due to malunion with osteopenia and blue sclerae were present. The patient was treated with z plasty of constriction band of the right tibia and bisphosphonate for osteogenesis imperfecta. This rare association of both collagen diseases may provide further insight for the pathogenesis of these diseases. 2015 BMJ Publishing Group Ltd.

  11. 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

  12. Amniotic epithelial cells: a new tool to combat aging and age-related diseases?

    Directory of Open Access Journals (Sweden)

    Clara Di Germanio

    2016-11-01

    Full Text Available The number of elderly people is growing at an unprecedented rate and this increase of the aging population is expected to have a direct impact on the incidence of age-related diseases and healthcare-associated costs. Thus, it is imperative that new tools are developed to fight and slow age-related diseases. Regenerative medicine is a promising strategy for the maintenance of health and function late in life; however, stem cell-based therapies face several challenges including rejection and tumor transformation. As an alternative, the placenta offers an extraordinary source of fetal stem cells, including the amniotic epithelial cells (AECs, which retain some of the characteristics of embryonic stem cells, but show low immunogenicity, together with immunomodulatory and anti-inflammatory activities. Because of these characteristics, AECs have been widely utilized in regenerative medicine. This perspective highlights different mechanisms triggered by transplanted AECs that could be potentially useful for anti-aging therapies, which include: Graft and differentiation for tissue regeneration in age-related settings, anti-inflammatory behavior to combat inflammaging, anti-tumor activity, direct lifespan and healthspan extension properties, and possibly rejuvenation in a manner reminiscent of heterochronic parabiosis.Here, we critically discuss benefits and limitation of AECs-based therapies in age-related diseases.

  13. Amniotic Epithelial Cells: A New Tool to Combat Aging and Age-Related Diseases?

    Science.gov (United States)

    Di Germanio, Clara; Bernier, Michel; de Cabo, Rafael; Barboni, Barbara

    2016-01-01

    The number of elderly people is growing at an unprecedented rate and this increase of the aging population is expected to have a direct impact on the incidence of age-related diseases and healthcare-associated costs. Thus, it is imperative that new tools are developed to fight and slow age-related diseases. Regenerative medicine is a promising strategy for the maintenance of health and function late in life; however, stem cell-based therapies face several challenges including rejection and tumor transformation. As an alternative, the placenta offers an extraordinary source of fetal stem cells, including the amniotic epithelial cells (AECs), which retain some of the characteristics of embryonic stem cells, but show low immunogenicity, together with immunomodulatory and anti-inflammatory activities. Because of these characteristics, AECs have been widely utilized in regenerative medicine. This perspective highlights different mechanisms triggered by transplanted AECs that could be potentially useful for anti-aging therapies, which include: Graft and differentiation for tissue regeneration in age-related settings, anti-inflammatory behavior to combat "inflammaging," anti-tumor activity, direct lifespan and healthspan extension properties, and possibly rejuvenation in a manner reminiscent of heterochronic parabiosis. Here, we critically discuss benefits and limitation of AECs-based therapies in age-related diseases.

  14. Fabrication of novel high surface area mushroom gilled fibers and their effects on human adipose derived stem cells under pulsatile fluid flow for tissue engineering applications.

    Science.gov (United States)

    Tuin, Stephen A; Pourdeyhimi, Behnam; Loboa, Elizabeth G

    2016-05-01

    The fabrication and characterization of novel high surface area hollow gilled fiber tissue engineering scaffolds via industrially relevant, scalable, repeatable, high speed, and economical nonwoven carding technology is described. Scaffolds were validated as tissue engineering scaffolds using human adipose derived stem cells (hASC) exposed to pulsatile fluid flow (PFF). The effects of fiber morphology on the proliferation and viability of hASC, as well as effects of varied magnitudes of shear stress applied via PFF on the expression of the early osteogenic gene marker runt related transcription factor 2 (RUNX2) were evaluated. Gilled fiber scaffolds led to a significant increase in proliferation of hASC after seven days in static culture, and exhibited fewer dead cells compared to pure PLA round fiber controls. Further, hASC-seeded scaffolds exposed to 3 and 6dyn/cm(2) resulted in significantly increased mRNA expression of RUNX2 after one hour of PFF in the absence of soluble osteogenic induction factors. This is the first study to describe a method for the fabrication of high surface area gilled fibers and scaffolds. The scalable manufacturing process and potential fabrication across multiple nonwoven and woven platforms makes them promising candidates for a variety of applications that require high surface area fibrous materials. We report here for the first time the successful fabrication of novel high surface area gilled fiber scaffolds for tissue engineering applications. Gilled fibers led to a significant increase in proliferation of human adipose derived stem cells after one week in culture, and a greater number of viable cells compared to round fiber controls. Further, in the absence of osteogenic induction factors, gilled fibers led to significantly increased mRNA expression of an early marker for osteogenesis after exposure to pulsatile fluid flow. This is the first study to describe gilled fiber fabrication and their potential for tissue engineering

  15. Accuracy of intraocular pressure by Tono-Pen XL over amniotic membrane patching in rabbits.

    Science.gov (United States)

    Kobayashi, Akira; Yoshita, Tsuyoshi; Shirao, Yutaka

    2003-04-01

    To report an accuracy of intraocular pressure (IOP) by Tono-Pen XL over amniotic membrane patching in rabbits. Experimental study. Seven male albino rabbits were used in this study. After general anesthesia, IOP was measured in both eyes using the Tono-Pen without amniotic membrane. A single layer amniotic membrane was then immediately placed over the rabbit eye, and the IOP measurement was repeated. The average IOP without amniotic membrane was 5.8 +/- 0.6 mm Hg (mean +/- SD). The average IOP through amniotic membrane was 6.0 +/- 0.4 mm Hg (mean +/- SD). There was no statistical difference in IOP measured on corneas with or without the amniotic membrane (P =.34). The accuracy of IOP measurement by Tono-Pen XL over a single layer of amniotic membrane patching was demonstrated using rabbit eyes. Further studies in human eyes may verify our findings in actual clinical settings.

  16. Congenital Amniotic Band Syndrome: Clinico-Epidemologic Study ...

    African Journals Online (AJOL)

    Congenital amniotic band syndrome (CABS) is an uncommon clinical entity with a global incidence of 1: 1, 200 to 1: 15, 000 live births. They nevertheless pose great functional, cosmetic and reconstructive challenge. There has been only one case report of this condition from our centre. This paper reviews the ...

  17. Short term results of pterygium surgery with adjunctive amniotic ...

    African Journals Online (AJOL)

    One patient developed dellen 1 week post‑operatively with complete resolution following conservative large soft contact lens application. Conclusion: Short term results suggests that adjunctive amniotic membrane transplant with pterygium excision is effective and safe. A larger randomized clinical trial with a longer ...

  18. 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 ...

  19. Amniotic band syndrom at Bobo Dioulasso university teaching ...

    African Journals Online (AJOL)

    ... a Z-plasty. The functional outcome was satisfactory with the acquisition of a plantar support for both children. Through these two observations, epidemiological, diagnostic, and particularities of the management of this condition are discussed in the Burkina-Faso. Key words: Amniotic band syndrom, surgery, birth defects ...

  20. Antibiotic Therapy Increases the Risk of Preterm Birth in Preterm Labor without Intra-Amniotic Microbes, but may Prolong the Gestation Period in Preterm Labor with Microbes, Evaluated by Rapid and High-Sensitive PCR System.

    Science.gov (United States)

    Yoneda, Satoshi; Shiozaki, Arihiro; Yoneda, Noriko; Ito, Mika; Shima, Tomoko; Fukuda, Kaori; Ueno, Tomohiro; Niimi, Hideki; Kitajima, Isao; Kigawa, Mika; Saito, Shigeru

    2016-04-01

    To examine the efficacy of the use of antibiotics in preterm labor (PTL) with intact membranes, after evaluating intra-amniotic microbes by our rapid and bacteria-free polymerase chain reaction (PCR) system. One hundred and four PTL patients before 32 weeks of gestation were recruited. Until 2012, antibiotics were empirically prescribed based on the clinical severity of PTL. Intra-amniotic microbes in stored samples were evaluated later by our newly established PCR system, and the efficacy of the use of antibiotics in PTL was evaluated. In the amniotic fluid (AF) microbe-negative patients (n = 67), antibiotic therapy significantly shortened the gestation period (P antibiotic therapy (proper antibiotic selection against identified AF microbes) was significantly associated with an increase in gestation period (P antibiotic therapy in PTL with intact membranes prolonged the gestation period. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  1. Maternal administration of solithromycin, a new, potent, broad-spectrum fluoroketolide antibiotic, achieves fetal and intra-amniotic antimicrobial protection in a pregnant sheep model.

    Science.gov (United States)

    Keelan, Jeffrey A; Kemp, Matthew W; Payne, Matthew S; Johnson, David; Stock, Sarah J; Saito, Masatoshi; Fernandes, Prabhavathi; Newnham, John P

    2014-01-01

    Solithromycin (CEM-101) is a new antibiotic that is highly potent against Ureaplasma and Mycoplasma spp. and active against many other antibiotic-resistant organisms. We have explored the maternal-amniotic-fetal pharmacokinetics of CEM-101 in a pregnant sheep model to assess its potential for treating intrauterine and antenatal infection. Chronically catheterized pregnant ewes (n = 6 or 7) received either a single maternal intravenous (i.v.) infusion of CEM-101 (10 mg/kg of body weight), a single intra-amniotic (i.a.) injection (1.4 mg/kg of estimated fetal weight), or a combined i.v. and i.a. dose. Maternal plasma (MP), fetal plasma (FP), and amniotic fluid (AF) samples were taken via catheter at intervals of 0 to 72 h postadministration, and concentrations of solithromycin and its bioactive polar metabolites (N-acetyl [NAc]-CEM-101 and CEM-214) were determined. Following maternal i.v. infusion, peak CEM-101 concentrations in MP, FP, and AF were 1,073, 353, and 214 ng/ml, respectively, representing a maternal-to-fetal plasma transfer efficiency of 34%. A single maternal dose resulted in effective concentrations (>30 ng/ml) in MP, FP, and AF sustained for >12 h. NAc-CEM-101 and CEM-214 exhibited delayed accumulation and clearance in FP and AF, resulting in an additive antimicrobial effect (>48 h). Intra-amniotic solithromycin injection resulted in elevated (∼50 μg/ml) and sustained CEM-101 concentrations in AF and significant levels in FP, although the efficiency of amniotic-to-fetal transfer was low (∼1.5%). Combined i.v. and i.a. administration resulted in primarily additive concentrations of CEM-101 in all three compartments. Our findings suggest that CEM-101 may provide, for the first time, an effective antimicrobial approach for the prevention and treatment of intrauterine infection and early prevention of preterm birth.

  2. Tissue engineering and the use of stem/progenitor cells for airway epithelium repair

    Directory of Open Access Journals (Sweden)

    GM Roomans

    2010-06-01

    Full Text Available Stem/progenitor cells can be used to repair defects in the airway wall, resulting from e.g., tumors, trauma, tissue reactions following long-time intubations, or diseases that are associated with epithelial damage. Several potential sources of cells for airway epithelium have been identified. These can be divided into two groups. The first group consists of endogenous progenitor cells present in the respiratory tract. This group can be subdivided according to location into (a a ductal cell type in the submucosal glands of the proximal trachea, (b basal cells in the intercartilaginous zones of the lower trachea and bronchi, (c variant Clara cells (Clarav-cells in the bronchioles and (d at the junctions between the bronchioles and the alveolar ducts, and (e alveolar type II cells. This classification of progenitor cell niches is, however, controversial. The second group consists of exogenous stem cells derived from other tissues in the body. This second group can be subdivided into: (a embryonic stem (ES cells, induced pluripotent stem (iPS cells, or amniotic fluid stem cells, (b side-population cells from bone marrow or epithelial stem cells present in bone marrow or circulation and (c fat-derived mesenchymal cells. Airway epithelial cells can be co-cultured in a system that includes a basal lamina equivalent, extracellular factors from mesenchymal fibroblasts, and in an air-liquid interface system. Recently, spheroid-based culture systems have been developed. Several clinical applications have been suggested: cystic fibrosis, acute respiratory distress syndrome, chronic obstructive lung disease, pulmonary fibrosis, pulmonary edema, and pulmonary hypertension. Clinical applications so far are few, but include subglottic stenosis, tracheomalacia, bronchiomalacia, and emphysema.

  3. Evolution of the snake body form reveals homoplasy in amniote Hox gene function.

    Science.gov (United States)

    Head, Jason J; Polly, P David

    2015-04-02

    Hox genes regulate regionalization of the axial skeleton in vertebrates, and changes in their expression have been proposed to be a fundamental mechanism driving the evolution of new body forms. The origin of the snake-like body form, with its deregionalized pre-cloacal axial skeleton, has been explained as either homogenization of Hox gene expression domains, or retention of standard vertebrate Hox domains with alteration of downstream expression that suppresses development of distinct regions. Both models assume a highly regionalized ancestor, but the extent of deregionalization of the primaxial domain (vertebrae, dorsal ribs) of the skeleton in snake-like body forms has never been analysed. Here we combine geometric morphometrics and maximum-likelihood analysis to show that the pre-cloacal primaxial domain of elongate, limb-reduced lizards and snakes is not deregionalized compared with limbed taxa, and that the phylogenetic structure of primaxial morphology in reptiles does not support a loss of regionalization in the evolution of snakes. We demonstrate that morphometric regional boundaries correspond to mapped gene expression domains in snakes, suggesting that their primaxial domain is patterned by a normally functional Hox code. Comparison of primaxial osteology in fossil and modern amniotes with Hox gene distributions within Amniota indicates that a functional, sequentially expressed Hox code patterned a subtle morphological gradient along the anterior-posterior axis in stem members of amniote clades and extant lizards, including snakes. The highly regionalized skeletons of extant archosaurs and mammals result from independent evolution in the Hox code and do not represent ancestral conditions for clades with snake-like body forms. The developmental origin of snakes is best explained by decoupling of the primaxial and abaxial domains and by increases in somite number, not by changes in the function of primaxial Hox genes.

  4. Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation

    Science.gov (United States)

    Riedel, Rodrigo; Pérez-Pérez, Antonio; Magatti, Marta; Maskin, Bernardo; Dueñas, José Luis; Parolini, Ornella; Sánchez-Margalet, Víctor; Varone, Cecilia L.

    2018-01-01

    Stem cells derived from placental tissues are an attractive source of cells for regenerative medicine. Amniotic epithelial cells isolated from human amnion (hAECs) have desirable and competitive characteristics that make them stand out between other stem cells. They have the ability to differentiate toward all three germ layers, they are not tumorigenic and they have immunosuppressive properties. Although liver transplantation is the best way to treat acute and chronic hepatic failure patients, there are several obstacles. Recently, stem cells have been spotlighted as alternative source of hepatocytes because of their potential for hepatogenic differentiation. In this work, we aimed to study the proliferation and survival of the hAECs during their hepatic differentiation. We have also analyzed the changes in pluripotency and hepatic markers. We differentiated amniotic cells applying a specific hepatic differentiation (HD) protocol. We determined by qRT-PCR that hAECs express significant levels of SOX-2, OCT-4 and NANOG during at least 15 days in culture and these pluripotent markers diminish during HD. SSEA-4 expression was reduced during HD, measured by immunofluorescence. Morphological characteristics became more similar to hepatic ones in differentiated cells and representative hepatic markers significantly augmented their expression, measured by qRT-PCR and Western blot. Cells achieved a differentiation efficiency of 75%. We observed that HD induced proliferation and promoted survival of hAECs, during 30 days in culture, evaluated by 3H-thymidine incorporation and MTT assay. HD also promoted changes in hAECs cell cycle. Cyclin D1 expression increased, while p21 and p53 levels were reduced. Immunofluorescence analysis showed that Ki-67 expression was upregulated during HD. Finally, ERK 1/2 phosphorylation, which is intimately linked to proliferation and cell survival, augmented during all HD process and the inhibition of this signaling pathway affected not only

  5. Conditioned medium derived from rat amniotic epithelial cells confers protection against inflammation, cancer, and senescence.

    Science.gov (United States)

    Di Germanio, Clara; Bernier, Michel; Petr, Michael; Mattioli, Mauro; Barboni, Barbara; de Cabo, Rafael

    2016-06-28

    Amniotic epithelial cells (AECs) are a class of fetal stem cells that derives from the epiblast and resides in the amnion until birth. AECs are suitable candidates for regenerative medicine because of the ease of collection, their low immunogenicity and inability to form tumors after transplantation. Even though human AECs have been widely investigated, the fact remains that very little is known about AECs isolated from rat, one of the most common animal models in medical testing. In this study, we showed that rat AECs retained stemness properties and plasticity, expressed the pluripotency markers Sox2, Nanog, and Oct4 and were able to differentiate toward the osteogenic lineage. The addition of conditioned medium collected from rat AECs to lipopolysaccharide-activated macrophages elicited anti-inflammatory properties through a decrease of Tnfa expression and slowed tumor cell proliferation in vitro and in vivo. The senescence-associated secretory phenotype was also significantly lower upon incubation of senescent human IMR-90 fibroblast cells with conditioned medium from rat AECs. These results confirm the potential of AECs in the modulation of inflammatory mechanisms and open new therapeutic possibilities for regenerative medicine and anti-aging therapies as well.

  6. Cell source-dependent in vivo immunosuppressive properties of mesenchymal stem cells derived from the bone marrow and synovial fluid of minipigs

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Won-Jae [College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Gyeongnam (Korea, Republic of); Hah, Young-Sool [Biomedical Research Institute, Gyeongsang National University Hospital, Jinju (Korea, Republic of); Ock, Sun-A. [Animal Biotechnology Division, National Institute of Animal Science, RDA, Suwon 441-706, Gyeonggi (Korea, Republic of); Lee, Jae-Hoon; Jeon, Ryong-Hoon; Park, Ji-Sung [College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Gyeongnam (Korea, Republic of); Lee, Sang-Il [Department of Internal Medicine and Institute of Health Sciences, Gyeongsang National University, Jinju (Korea, Republic of); Rho, Na-Young [Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada N1G 4S7 (Canada); Rho, Gyu-Jin [College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Gyeongnam (Korea, Republic of); Research Institute of Life Sciences, Gyeongsang National University, Jinju 660-701, Gyeongnam (Korea, Republic of); Lee, Sung-Lim, E-mail: sllee@gnu.ac.kr [College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Gyeongnam (Korea, Republic of); Research Institute of Life Sciences, Gyeongsang National University, Jinju 660-701, Gyeongnam (Korea, Republic of)

    2015-05-01

    The in vitro differentiation and immunosuppressive capacity of mesenchymal stem cells (MSCs) derived from synovial fluid (SF-MSCs) and bone marrow extract (BM-MSCs) in an isogenic background of minipigs were comparatively analyzed in a collagen-induced arthritis (CIA) mouse model of rheumatoid arthritis (RA). The proliferation capacity and expression of pluripotent transcription factors (Oct3/4 and Sox2) were significantly (P<0.05) higher in SF-MSCs than in BM-MSCs. The differentiation capacity of SF-MSCs into adipocytes, osteocytes and neurocytes was significantly (P<0.05) lower than that of BM-MSCs, and the differentiation capacity of SF-MSCs into chondrocytes was significantly (P<0.05) higher than that of BM-MSCs. Systemic injection of BM- and SF-MSCs significantly (P<0.05) ameliorated the clinical symptoms of CIA mice, with SF-MSCs having significantly (P<0.05) higher clinical and histopathological recovery scores than BM-MSCs. Furthermore, the immunosuppressive properties of SF-MSCs in CIA mice were associated with increased levels of the anti-inflammatory cytokine interleukin (IL)-10, and decreased levels of the pro-inflammatory cytokine IL-1β and osteoclast-related sRANKL. In conclusion, SF-MSCs exhibited eminent pluripotency and differentiation capacity into chondrocytes, addition to substantial in vivo immunosuppressive capacity by elevating IL-10 and reducing IL-1β levels in CIA mice. - Highlights: • Immunosuppressive capacity of BM-, SM-, and SF-MSCs was evaluated in an RA model. • Proliferation, pluripotency and chondrogenic differentiation capacity were higher in SF-MSCs. • SF-MSCs exhibited improved therapeutic effects than BM-MSCs. • SF-MSCs may have applications as immunosuppressive therapy in autoimmune diseases.

  7. Effect of arthritic synovial fluids on the expression of immunomodulatory factors by mesenchymal stem cells: an explorative in-vitro study

    Directory of Open Access Journals (Sweden)

    Maarten J.C. Leijs

    2012-08-01

    Full Text Available Background: In diseased joints, the catabolic environment results in progressive joint damage. Mesenchymal stem cells (MSCs can have immunomodulatory effect by secreting anti-inflammatory factors. To exert these effects, MSCs need to be triggered by pro-inflammatory cytokines. To explore the potential of MSCs as a treatment for diseased joints, we studied the effect of synovial fluid (SF from donors with different joint diseases and donors without joint pathology on the immunomodulatory capacities of human MSCs in vitro. We hypothesized that SF of diseased joints influences the immunomodulatory effects of MSCs. Materials & Methods: MSCs were cultured in medium with SF of six osteoarthritis (OA or six rheumatoid arthritis (RA donors and three donors without joint pathology were used as control. Gene expressions of IL-6, HGF, TNFa, TGFb1 and indoleamine 2,3-dioxygenase (IDO were analysed. L-kynurenine concentration in conditioned medium (CM by MSCs with SF was determined as a measure of IDO activity by MSCs. Furthermore, the effect of CM with SF on proliferation of activated lymphocytes was analysed. Results: Addition of SF significantly up-regulated the mRNA expression of IL-6 and IDO in MSCs. SF(OA induced significantly higher expression of IDO than SF(control, although no difference in IDO activity of the MSCs could be shown with a L-kynurenine assay. Medium conditioned by MSCs with SF(OA or RA suppressed activated lymphocyte proliferation in vitro more than medium conditioned by MSCs without SF or with SF(control.Discussion: SF can influence the expression of genes involved in immunomodulation by MSCs and the effect on lymphocyte proliferation. We found indications for disease-specific differences between SFs but the variation between donors, even within one disease group was high. These data warrant further research to examine the potential application of MSC therapy in arthritic joints.

  8. Human Amniotic Membrane-Derived Products in Sports Medicine: Basic Science, Early Results, and Potential Clinical Applications.

    Science.gov (United States)

    Riboh, Jonathan C; Saltzman, Bryan M; Yanke, Adam B; Cole, Brian J

    2016-09-01

    Amniotic membrane (AM)-derived products have been successfully used in ophthalmology, plastic surgery, and wound care, but little is known about their potential applications in orthopaedic sports medicine. To provide an updated review of the basic science and preclinical and clinical data supporting the use of AM-derived products and to review their current applications in sports medicine. Systematic review. A systematic search of the literature was conducted using the Medline, EMBASE, and Cochrane databases. The search term amniotic membrane was used alone and in conjunction with stem cell, orthopaedic, tissue engineering, scaffold, and sports medicine. The search identified 6870 articles, 80 of which, after screening of the titles and abstracts, were considered relevant to this study. Fifty-five articles described the anatomy, basic science, and nonorthopaedic applications of AM-derived products. Twenty-five articles described preclinical and clinical trials of AM-derived products for orthopaedic sports medicine. Because the level of evidence obtained from this search was not adequate for systematic review or meta-analysis, a current concepts review on the anatomy, physiology, and clinical uses of AM-derived products is presented. Amniotic membranes have many promising applications in sports medicine. They are a source of pluripotent cells, highly organized collagen, antifibrotic and anti-inflammatory cytokines, immunomodulators, and matrix proteins. These properties may make it beneficial when applied as tissue engineering scaffolds, improving tissue organization in healing, and treatment of the arthritic joint. The current body of evidence in sports medicine is heavily biased toward in vitro and animal studies, with little to no human clinical data. Nonetheless, 14 companies or distributors offer commercial AM products. The preparation and formulation of these products alter their biological and mechanical properties, and a thorough understanding of these

  9. Comparison of human mesenchymal stromal cells from four neonatal tissues: Amniotic membrane, chorionic membrane, placental decidua and umbilical cord.

    Science.gov (United States)

    Araújo, Anelise Bergmann; Salton, Gabrielle Dias; Furlan, Juliana Monteiro; Schneider, Natália; Angeli, Melissa Helena; Laureano, Álvaro Macedo; Silla, Lúcia; Passos, Eduardo Pandolfi; Paz, Ana Helena

    2017-05-01

    Mesenchymal stromal cells (MSCs) are being investigated as a potential alternative for cellular therapy. This study was designed to compare the biological characteristics of MSCs isolated from amniotic membrane (A-MSCs), chorionic membrane (C-MSCs), placental decidua (D-MSCs) and umbilical cord (UC-MSCs) to ascertain whether any one of these sources is superior to the others for cellular therapy purposes. MSCs were isolated from amniotic membrane, chorionic membrane, umbilical cord and placental decidua. Immunophenotype, differentiation ability, cell size, cell complexity, polarity index and growth kinetics of MSCs isolated from these four sources were analyzed. MSCs were successfully isolated from all four sources. Surface marker profile and differentiation ability were consistent with human MSCs. C-MSCs in suspension were the smallest cells, whereas UC-MSCs presented the greatest length and least width. A-MSCs had the lowest polarity index and UC-MSCs, as more elongated cells, the highest. C-MSCs, D-MSCs and UC-MSCs exhibited similar growth capacity until passage 8 (P8); C-MSCs presented better lifespan, whereas insignificant proliferation was observed in A-MSCs. Neonatal and maternal tissues can serve as sources of multipotent stem cells. Some characteristics of MSCs obtained from four neonatal tissues were compared and differences were observed. Amniotic membrane was the least useful source of MSCs, whereas chorionic membrane and umbilical cord were considered good options for future use in cell therapy because of the known advantages of immature cells. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  10. Amniotic membrane transplantation for ligneous conjunctivitis in a doberman.

    Science.gov (United States)

    O'Leary, Louise; Specht, Andrew; Isaza, Natalie; Kallberg, Maria; Barrie, Kathleen; Cottrell, Deborah; Plummer, Caryn

    2018-02-26

    An approximately 2-year-old female Doberman Pinscher was referred for the evaluation of bilateral, chronic proliferative conjunctivitis. Ophthalmic examination revealed bilateral thick, opaque pseudomembranes originating from the conjunctivae that prevented visualization of the cornea and interior structures of the eye. Histopathological findings of biopsies of the pseudomembranes were consistent with ligneous conjunctivitis. Serum plasminogen activity levels were within the normal range. Treatment with topical and systemic anti-inflammatory and immunosuppressive drugs did not improve the conjunctival lesions. The pseudomembranes were surgically excised, and the conjunctival surfaces were reconstructed with amniotic membrane. At final re-examination two years postsurgery, there was no evidence of recurrence of the pseudomembranes. To the authors' knowledge, this is the first reported case of the successful treatment of canine ligneous conjunctivitis with amniotic membrane transplantation. © 2018 American College of Veterinary Ophthalmologists.

  11. Photo-cross-linking of amniotic membranes for limbal epithelial cell cultivation

    Energy Technology Data Exchange (ETDEWEB)

    Lai, Jui-Yang, E-mail: jylai@mail.cgu.edu.tw

    2014-12-01

    In the present study, we developed photo-cross-linked amniotic membrane (AM) as a limbal stem cell niche. After ultraviolet (UV) irradiation for varying time periods, the biological tissues were studied by determinations of cross-linking structure, degradability, and nutrient permeation ability. Our results showed that the number of cross-links per unit mass of AM significantly increased with increasing illumination time from 5 to 50 min. However, the cross-link formation was inhibited by longer irradiation time (i.e., 150 min), probably due to the scission of tissue collagen chains through irradiation. The biological stability and matrix permeability of photo-cross-linked AM materials strongly depended on their cross-linking densities affected by the UV irradiation. In vitro biocompatibility studies including cell viability and pro-inflammatory gene expression analyses demonstrated that, irrespective of the irradiation time employed, the physically cross-linked biological tissues exhibited negligible cytotoxicity and similar interleukin-6 (IL-6) mRNA levels. The data clearly indicate that these AM matrices do not cause potential harm to the corneal epithelial cells. After the growth of limbal epithelial cells (LECs) on AM substrates, Western blot analyses were conducted to examine the expression of ABCG2. It was found that the ability of UV-irradiated AM to maintain the undifferentiated precursor cell phenotype was significantly enhanced with increasing extent of photo-cross-linking. In summary, the UV irradiation time may have a profound influence on the fabrication of photo-cross-linked AM matrices for LEC cultivation. - Highlights: • We report the development of photo-cross-linked AM as a limbal stem cell niche. • Cross-linked structure of tissue materials was controlled by UV irradiation time. • Biostability and matrix permeability of AM depended on cross-linking density. • All the studied photo-cross-linked AM showed good in vitro biocompatibility.

  12. Limbal Stem Cell Deficiency and Treatment with Stem Cell Transplantation.

    Science.gov (United States)

    Barut Selver, Özlem; Yağcı, Ayşe; Eğrilmez, Sait; Gürdal, Mehmet; Palamar, Melis; Çavuşoğlu, Türker; Ateş, Utku; Veral, Ali; Güven, Çağrı; Wolosin, Jose Mario

    2017-10-01

    The cornea is the outermost tissue of the eye and it must be transparent for the maintenance of good visual function. The superficial epithelium of the cornea, which is renewed continuously by corneal stem cells, plays a critical role in the permanence of this transparency. These stem cells are localized at the cornea-conjunctival transition zone, referred to as the limbus. When this zone is affected/destroyed, limbal stem cell deficiency ensues. Loss of limbal stem cell function allows colonization of the corneal surface by conjunctival epithelium. Over 6 million people worldwide are affected by corneal blindness, and limbal stem cell deficiency is one of the main causes. Fortunately, it is becoming possible to recover vision by autologous transplantation of limbal cells obtained from the contralateral eye in unilateral cases. Due to the potential risks to the donor eye, only a small amount of tissue can be obtained, in which only 1-2% of the limbal epithelial cells are actually limbal stem cells. Vigorous attempts are being made to expand limbal stem cells in culture to preserve or even enrich the stem cell population. Ex vivo expanded limbal stem cell treatment in limbal stem cell deficiency was first reported in 1997. In the 20 years since, various protocols have been developed for the cultivation of limbal epithelial cells. It is still not clear which method promotes effective stem cell viability and this remains a subject of ongoing research. The most preferred technique for limbal cell culture is the explant culture model. In this approach, a small donor eye limbal biopsy is placed as an explant onto a biocompatible substrate (preferably human amniotic membrane) for expansion. The outgrowth (cultivated limbal epithelial cells) is then surgically transferred to the recipient eye. Due to changing regulations concerning cell-based therapy, the implementation of cultivated limbal epithelial transplantation in accordance with Good Laboratory Practice using

  13. Limbal Stem Cell Deficiency and Treatment with Stem Cell Transplantation

    Directory of Open Access Journals (Sweden)

    Özlem Barut Selver

    2017-12-01

    Full Text Available The cornea is the outermost tissue of the eye and it must be transparent for the maintenance of good visual function. The superficial epithelium of the cornea, which is renewed continuously by corneal stem cells, plays a critical role in the permanence of this transparency. These stem cells are localized at the cornea-conjunctival transition zone, referred to as the limbus. When this zone is affected/destroyed, limbal stem cell deficiency ensues. Loss of limbal stem cell function allows colonization of the corneal surface by conjunctival epithelium. Over 6 million people worldwide are affected by corneal blindness, and limbal stem cell deficiency is one of the main causes. Fortunately, it is becoming possible to recover vision by autologous transplantation of limbal cells obtained from the contralateral eye in unilateral cases. Due to the potential risks to the donor eye, only a small amount of tissue can be obtained, in which only 1-2% of the limbal epithelial cells are actually limbal stem cells. Vigorous attempts are being made to expand limbal stem cells in culture to preserve or even enrich the stem cell population. Ex vivo expanded limbal stem cell treatment in limbal stem cell deficiency was first reported in 1997. In the 20 years since, various protocols have been developed for the cultivation of limbal epithelial cells. It is still not clear which method promotes effective stem cell viability and this remains a subject of ongoing research. The most preferred technique for limbal cell culture is the explant culture model. In this approach, a small donor eye limbal biopsy is placed as an explant onto a biocompatible substrate (preferably human amniotic membrane for expansion. The outgrowth (cultivated limbal epithelial cells is then surgically transferred to the recipient eye. Due to changing regulations concerning cell-based therapy, the implementation of cultivated limbal epithelial transplantation in accordance with Good Laboratory

  14. Responses of Preterm Pigs to an Oral Fluid Supplement During Parenteral Nutrition

    DEFF Research Database (Denmark)

    Berding, Kirsten; Makarem, Patty; Hance, Brittany

    2016-01-01

    Background: Nutrients and electrolytes in amniotic fluid swallowed by fetuses are important for growth and development. Yet, preterm infants requiring parenteral nutrition (PN) receive minimal or no oral inputs. With the limited availability of amniotic fluid, we evaluated the responses of preterm...... enterally (n = 10) or intravenously (n = 8). Outcome measures after 96 hours were weight gain, blood chemistry, organ weights, and small intestine mass and brush-border membrane carbohydrases. Results: The OFS did not improve weight gain compared with providing lactated Ringer’s orally or intravenously...

  15. 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

    Insulin-like growth factor-I (IGF-I) and non-esterified fatty acids (NEFA) play an essential role in fetal growth and development. To date, fetal fluids IGF-I and NEFA levels at term canine pregnancy are unknown and could be related to the neonatal development and breed size. For these reasons, the aims of the present study were as follows: (1) to evaluate IGF-I and NEFA concentrations in fetal fluids collected from normally developed and viable newborn puppies born at term of normal pregnancies; (2) to assess possible differences between IGF-I and NEFA levels in amniotic compared with allantoic fluid; (3) to detect possible relationship between breed body size and IGF-I and NEFA amniotic and allantoic concentrations; (4) to evaluate possible differences in IGF-I fetal fluids levels between male and female puppies; and (5) to assess possible correlations between the two hormones in each type of fluid. The study enrolled 25 pure breed bitches submitted to elective Cesarean section at term because of the high risk of dystocia or previous troubles at parturition. At surgery, amniotic and allantoic fluids were collected and assayed for IGF-I and NEFA. IGF-I and NEFA amounts in both amniotic and allantoic fluids of different breed size bitches (small: ≤10 kg; medium: 11-25 kg; large: 26-40 kg) were detected, as well as the effect of gender on IGF-I levels. On a total of 73 amniotic and 76 allantoic samples collected by normal, viable, and mature newborns, the mean IGF-I concentration was significantly higher in amniotic than in allantoic fluid in all three groups, but the amniotic IGF-I levels were significantly lower in small and medium size bitches when compared with large ones. No significant differences were found in allantoic IGF-I concentrations among size groups. A significant effect of the puppy gender on IGF-I content in both fetal fluids was not reported. Regarding NEFA, in all the three groups, the mean NEFA concentration did not significantly differ

  16. Effect of placenta previa on neonatal respiratory disorders and amniotic lamellar body counts at 36-38weeks of gestation.

    Science.gov (United States)

    Tsuda, Hiroyuki; Kotani, Tomomi; Sumigama, Seiji; Mano, Yukio; Hua, Li; Hayakawa, Hiromi; Hayakawa, Masahiro; Sato, Yoshiaki; Kikkawa, Fumitaka

    2014-01-01

    Pregnancies with placenta previa are significantly associated with preterm delivery and cesarean section. Therefore particular attention should be paid to the incidence of neonatal respiratory disorders in pregnancies with placenta previa. The purpose of this study is to examine the relationship between placenta previa and neonatal respiratory disorders, including respiratory distress syndrome (RDS) and transient tachypnea of the newborn (TTN), and to evaluate the impact of placenta previa on the amniotic lamellar body count (LBC) values. We analyzed the data from 186 registered elective cesarean cases without fetal or maternal complications at 36-38weeks of gestation. Amniotic fluid samples were analyzed immediately without centrifugation, and the LBC was measured using a platelet channel on the Sysmex XE-2100. RDS was present in four neonates (2.2%) and TTN in 12 neonates (6.5%). The rate of TTN was significantly higher and the LBC values were significantly lower in the placenta previa group than in the control group (P=0.002 and P=0.024). The adjusted odds ratio for neonatal TTN was 7.20 (95% confidence interval: 6.58-7.88) among females with placenta previa. In placenta previa, warning bleeding was a significant factor protecting against neonatal respiratory disorders (P=0.046). Placenta previa in itself is a risk factor for neonatal TTN. When an elective cesarean section is performed in cases with uncomplicated placenta previa, special care should be taken to monitor for neonatal TTN even at 36-38weeks of gestation. © 2013.

  17. Outcomes following intra-amniotic instillation with indigo carmine to diagnose prelabor rupture of membranes in singleton pregnancies: a single center experience.

    Science.gov (United States)

    Adekola, Henry; Gill, Navleen; Sakr, Sharif; Hobson, Deslyn; Bryant, David; Abramowicz, Jacques S; Soto, Eleazar

    2016-01-01

    To evaluate clinical outcomes of women with singleton pregnancies that underwent intra-amniotic dye instillation (amniodye test) following equivocal diagnosis of prelabor rupture of membranes (PROM). Records of 34 pregnant women who underwent amniodye test for equivocal PROM were reviewed. Comparisons of characteristics, amniotic fluid (AF) cultures, AF interleukin (IL)-6 concentrations, and placenta pathology results between women who tested positive and those who tested negative were performed. A sub-analysis of women who were amniodye test-negative was also performed. (1) Commonest indication for amniodye test was a typical history of PROM with positive conventional tests and persistently normal AF volume, (2) amniodye test-positive women had a shorter procedure-to-delivery interval (p = 0.008), and a greater proportion of histologic acute chorioamnionitis (p = 0.04) and funisitis (p = 0.01) than amniodye-negative women, and (3) in addition to similarities to women with amniodye-positive test, amniodye test-negative women who delivered <34 weeks, had a greater proportion of women with risk for preterm birth (p = 0.04), than their counterparts who delivered between 34 0/7 and 36 6/7 weeks. Equivocal diagnosis of PPROM should warrant an amniodye test to avoid iatrogenic intervention in women with intact amniotic membranes. AF analysis should be performed in amniodye test-negative women.

  18. Reliability of intraocular pressure by Tono-Pen XL over amniotic membrane patch in human.

    Science.gov (United States)

    Yoshita, Tsuyoshi; Kobayashi, Akira; Takahashi, Mami; Sugiyama, Kazuhisa

    2004-10-01

    To determine the reliability of Tono-Pen XL in measuring intraocular pressure (IOP) over the amniotic membrane patch in human eyes. Ten healthy volunteers (nine males and one female) participated in this study. After topical anesthesia, IOP was measured using the Tono-Pen XL before and after the application of a single-layer amniotic membrane patch in the right eye and double-layer amniotic membrane patch in the left eye. We found no statistical difference between the IOP measured with and without a single layer amniotic membrane (P = 0.808). Nor did we find any statistical difference between the IOP measured with and without double-layer amniotic membrane patch (P = 0.813). The average thickness of the single- and the double-layer amniotic membrane patch measured by pachymetry was 131.9 +/- 31.4 microm (mean +/- SD) and was 246.1 +/- 69.3 microm (mean +/- SD), respectively. This study showed that it is reliable to measure IOP by Tono-Pen XL over single or a double layer of amniotic membrane patch on healthy human eyes. Further studies are required to determine the accuracy of IOP measurement over amniotic membrane patch on diseased corneas.

  19. Identification of emergent motion compartments in the amniote embryo.

    Science.gov (United States)

    Loganathan, Rajprasad; Little, Charles D; Joshi, Pranav; Filla, Michael B; Cheuvront, Tracey J; Lansford, Rusty; Rongish, Brenda J

    2014-01-01

    The tissue scale deformations (≥ 1 mm) required to form an amniote embryo are poorly understood. Here, we studied ∼400 μm-sized explant units from gastrulating quail embryos. The explants deformed in a reproducible manner when grown using a novel vitelline membrane-based culture method. Time-lapse recordings of latent embryonic motion patterns were analyzed after disk-shaped tissue explants were excised from three specific regions near the primitive streak: 1) anterolateral epiblast, 2) posterolateral epiblast, and 3) the avian organizer (Hensen's node). The explants were cultured for 8 hours-an interval equivalent to gastrulation. Both the anterolateral and the posterolateral epiblastic explants engaged in concentric radial/centrifugal tissue expansion. In sharp contrast, Hensen's node explants displayed Cartesian-like, elongated, bipolar deformations-a pattern reminiscent of axis elongation. Time-lapse analysis of explant tissue motion patterns indicated that both cellular motility and extracellular matrix fiber (tissue) remodeling take place during the observed morphogenetic deformations. As expected, treatment of tissue explants with a selective Rho-Kinase (p160ROCK) signaling inhibitor, Y27632, completely arrested all morphogenetic movements. Microsurgical experiments revealed that lateral epiblastic tissue was dispensable for the generation of an elongated midline axis- provided that an intact organizer (node) is present. Our computational analyses suggest the possibility of delineating tissue-scale morphogenetic movements at anatomically discrete locations in the embryo. Further, tissue deformation patterns, as well as the mechanical state of the tissue, require normal actomyosin function. We conclude that amniote embryos contain tissue-scale, regionalized morphogenetic motion generators, which can be assessed using our novel computational time-lapse imaging approach. These data and future studies-using explants excised from overlapping anatomical

  20. Effect of amniotic membrane to reduce postlaminectomy epidural adhesion on a rat model.

    Science.gov (United States)

    Choi, Hyu Jin; Kim, Kyoung Beom; Kwon, Young-Min

    2011-06-01

    Epidural fibrosis and adhesion are the main reasons for post-laminectomy sustained pain and functional disability. In this study, the authors investigate the effect of irradiated freeze-dried human amniotic membrane on reducing epidural adhesion after laminectomy on a rat model. A total of 20 rats were divided into two groups. The group A did not receive human amniotic membrane implantation after laminectomy and group B underwent human amniotic membrane implantation after laminectomy. Gross and microscopic findings were evaluated and compared at postoperative 1, 3 and 8 weeks. The amount of scar tissue and tenacity were reduced grossly in group of rats with human amniotic membrane implantation (group B). On a microscopic evaluation, there were less inflammatory cell infiltration and fibroblast proliferation in group B. This experimental study shows that implantation of irradiated freeze-dried human amniotic membrane reduce epidural fibrosis and adhesion after spinal laminectomy in a rat model.

  1. Transplantation of amniotic membrane to the subretinal space in pigs

    DEFF Research Database (Denmark)

    Kiilgaard, Jens Folke; Scherfig, Erik; Prause, Jan Ulrik

    2012-01-01

    Purpose. To investigate the effect of transplanted amniotic membrane (AM) on subretinal wound healing. Methods. Nine Danish Landrace pigs had surgical removal of retinal pigment epithelium (RPE) and mechanical damage of Bruch's membrane (BM) and served as a control group. 15 pigs additionally had...... is well tolerated in the subretinal space, causes only limited inflammation, and is covered with a monolayer of pigmented cells when in contact with the host RPE. Conclusions. AM modifies choroidal neovascularisation after BM damage and may serve as a basement membrane substitute for the RPE....

  2. Stem Cells

    Science.gov (United States)

    Stem cells are cells with the potential to develop into many different types of cells in the body. They serve as a repair ... body. There are two main types of stem cells: embryonic stem cells and adult stem cells. Stem ...

  3. Human amniotic mesenchymal stromal cell transplantation improves endometrial regeneration in rodent models of intrauterine adhesions.

    Science.gov (United States)

    Gan, Lu; Duan, Hua; Xu, Qian; Tang, Yi-Qun; Li, Jin-Jiao; Sun, Fu-Qing; Wang, Sha

    2017-05-01

    Intrauterine adhesion (IUA) is a common uterine cavity disease characterized by the unsatisfactory regeneration of damaged endometria. Recently, stem cell transplantation has been proposed to promote the recovery process. Here we investigated whether human amniotic mesenchymal stromal cells (hAMSCs), a valuable resource for transplantation therapy, could improve endometrial regeneration in rodent IUA models. Forty female Sprague-Dawley rats were randomly assigned to five groups: normal, sham-operated, mechanical injury, hAMSC transplantation, and negative control group. One week after intervention and transplantation, histological analyses were performed, and immunofluorescent and immunohistochemical expression of cell-specific markers and messenger RNA expression of cytokines were measured. Thicker endometria, increased gland numbers and fewer fibrotic areas were found in the hAMSC transplantation group compared with the mechanical injury group. Engraftment of hAMSCs was detected by the presence of anti-human nuclear antigen-positive cells in the endometrial glands of the transplantation uteri. Transplantation of hAMSCs significantly decreased messenger RNA levels of pro-inflammatory cytokines (tumor necrosis factor-α and interleukin-1β), and increased those of anti-inflammatory cytokines (basic fibroblast growth factor, and interleukin-6) compared with the injured uterine horns. Immunohistochemical expression of endometrial epithelial cells was revealed in specimens after hAMSC transplantation, whereas it was absent in the mechanically injured uteri. hAMSC transplantation promotes endometrial regeneration after injury in IUA rat models, possibly due to immunomodulatory properties. These cells provide a more easily accessible source of stem cells for future research into the impact of cell transplantation on damaged endometria. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  4. A skin substitute based on human amniotic membrane.

    Science.gov (United States)

    Tauzin, Hélène; Rolin, Gwenaël; Viennet, Céline; Saas, Philippe; Humbert, Philippe; Muret, Patrice

    2014-06-01

    Human amniotic membrane (HAM) has biological properties which are useful for wound healing. HAM is notably one of the therapeutic alternatives for venous leg ulcer care. Indeed, a prospective clinical study has demonstrated that cryopreserved HAM transplantation for leg ulcer is feasible, safe and has beneficial effects: 80 % of the patients had a significant clinical response. Nevertheless, at the end of the 3-month follow-up period, only 20 % of the ulcers were totally closed. The aim of this work was to create and characterize a model of epidermized HAM. The method of HAM desepithelialization was validated by histology, immunohistochemistry and scanning electron microscopy. Then, de-epithelialized HAM was seeded with primary keratinocytes. After 21 days of culture, 15 at the air-liquid interface, the model obtained was analyzed histologically and by immunohistochemistry. The amniotic basement membrane was preserved during enzymatic desepithelialization of HAM. Primary keratinocytes proliferated on HAM: the model obtained showed involucrin expression and had a good basement membrane. As re-epithelialization is an important step for ulcer closure, a model of epidermized HAM could be used to speed up the healing of such wounds.

  5. Radioimmunoassay of the myelin basic protein in biological fluids, conditions improving sensitivity and specificity

    International Nuclear Information System (INIS)

    Delassalle, A.; Jacque, C.; Raoul, M.; Legrand, J.C.; Cesselin, F.; Drouet, J.

    1980-01-01

    The radioimmunoassay (RIA) for myelin basic protein (MBP) in biological fluids was reassessed in order to improve its sensitivity and eliminate some interferences. By using the pre-incubation technique and the charcoal-dextram-horse serum mixture for the separation step, the detection limit could be lowered to 200 pg/ml for cerebrospinal fluids (CSF), amniotic fluids (AF) and nervous tissue extracts and 600 pg/ml for sera. The RIA could be used directly on CSF, AF and nervous tissue extracts. Sera, however, had to be heated in citrate buffer at 100 0 C in order to discard interfering material. The present method is 10 to 20 times more sensitive than others previously published. Moreover, it can be applied to amniotic fluid. The biological fluids had to be promptly frozen to avoid degradation of MBP

  6. In vivo analysis of stromal integration of multilayer amniotic membrane transplantation in corneal ulcers.

    Science.gov (United States)

    Nubile, Mario; Dua, Harminder S; Lanzini, Manuela; Ciancaglini, Marco; Calienno, Roberta; Said, Dalia G; Pocobelli, Augusto; Mastropasqua, Rodolfo; Carpineto, Paolo

    2011-05-01

    To evaluate integration of amniotic membrane into the corneal stroma using laser scanning in vivo confocal microscopy and anterior segment optical coherence tomography (AS-OCT). Prospective noncomparative interventional case series. Twenty-two eyes of 22 consecutive patients (mean age 53.9 ± 9.2 years) presenting with noninfectious corneal ulcers and stromal thinning unresponsive to medical treatment were enrolled. Multiple layers of amniotic membrane were applied over the ulcer bed to fill the ulcer crater and held in place with an overlying amniotic membrane patch, which was anchored to the surrounding cornea with 10-0 nylon interrupted sutures. Outcome measures were healing of the corneal ulcers, corneal morphology and stromal thickness changes at the ulcer site as measured by AS-OCT and surface epithelialization, stromal repopulation, and structural modifications of the amniotic membrane grafts as evaluated by confocal microscopy. Follow-up extended to 12 months. Successful result was observed in 20 of 22 eyes (90.9%). AS-OCT showed that the mean residual stromal thickness at the ulcer bed was 222 ± 70 μm before surgery. The mean thickness of amniotic membrane layers at the same site was 394 ± 80 μm while the mean total corneal thickness was 623 ± 51 μm at day 1 post surgery. Thereafter a progressive reduction in thickness to 420 ± 61 μm at 6 months occurred, after which the thickness stabilized. Confocal microscopy showed that integration of the amniotic membrane tissues with corneal stroma was preceded by epithelialization over the amniotic membrane covering the ulcer. This occurred 15 ± 5 days post surgery in the successful cases. Confocal microscopy also showed that the amniotic membrane patch was degraded during the first few weeks after surgery, while the integrated amniotic tissues underwent progressive modifications characterized by early loss of amniotic epithelial cells, changes in fibrillar structure, and migration into the amniotic stroma

  7. Fluid shear stress primes mouse embryonic stem cells for differentiation in a self-renewing environment via heparan sulfate proteoglycans transduction.

    Science.gov (United States)

    Toh, Yi-Chin; Voldman, Joel

    2011-04-01

    Shear stress is a ubiquitous environmental cue experienced by stem cells when they are being differentiated or expanded in perfusion cultures. However, its role in modulating self-renewing stem cell phenotypes is unclear, since shear is usually only studied in the context of cardiovascular differentiation. We used a multiplex microfluidic array, which overcomes the limitations of macroperfusion systems in shear application throughput and precision, to initiate a comprehensive, quantitative study of shear effects on self-renewing mouse embryonic stem cells (mESCs), where shear stresses varying by >1000 times (0.016-16 dyn/cm(2)) are applied simultaneously. When compared with static controls in the presence or absence of a saturated soluble environment (i.e., mESC-conditioned medium), we ascertained that flow-induced shear stress specifically up-regulates the epiblast marker Fgf5. Epiblast-state transition in mESCs involves heparan sulfate proteoglycans (HSPGs), which have also been shown to transduce shear stress in endothelial cells. By disrupting (with sulfation inhibitors and heparinase) and partially reconstituting (with heparin) HSPG function, we show that mESCs also mechanically sense shear stress via HSPGs to modulate Fgf5 expression. This study demonstrates that self-renewing mESCs possess the molecular machinery to sense shear stress and provides quantitative shear application benchmarks for future scalable stem cell culture systems.

  8. The fluid dynamics of human birth

    Science.gov (United States)

    Lehn, Andrea; Leftwich, Megan C.

    2012-11-01

    This study investigates the fluid dynamics associated with the human birth process. Specifically, we investigate the role of the viscosity of the amniotic fluid in transferring force from the contracting uterus to the fetus during delivery. This experimental work uses an approximate uterus and dilated cervix-fabricated with liquid latex-filled with a fluid of known viscosity and an oblong solid fetus. The force required to extract the fetus is recorded for several values of amniotic viscosity. The study looks at both pull-out force values (where the fetus is pulled from outside the uterus) and push-out force values (where pressure in the experimental uterus is used to remove the fetus). In addition to the viscosity study, we also investigate the increased force required to deliver an offset fetus by tilting the major axis of the oblong fetus and repeating the pull-and push-out experiments. This study will provide knowledge about the fundamental fluid dynamic processes involved in human birth.

  9. 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

  10. 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

  11. Sex Chromosome Evolution in Amniotes: Applications for Bacterial Artificial Chromosome Libraries

    Directory of Open Access Journals (Sweden)

    Daniel E. Janes

    2011-01-01

    Full Text Available Variability among sex chromosome pairs in amniotes denotes a dynamic history. Since amniotes diverged from a common ancestor, their sex chromosome pairs and, more broadly, sex-determining mechanisms have changed reversibly and frequently. These changes have been studied and characterized through the use of many tools and experimental approaches but perhaps most effectively through applications for bacterial artificial chromosome (BAC libraries. Individual BAC clones carry 100–200 kb of sequence from one individual of a target species that can be isolated by screening, mapped onto karyotypes, and sequenced. With these techniques, researchers have identified differences and similarities in sex chromosome content and organization across amniotes and have addressed hypotheses regarding the frequency and direction of past changes. Here, we review studies of sex chromosome evolution in amniotes and the ways in which the field of research has been affected by the advent of BAC libraries.

  12. Prompt versus delayed amniotic membrane application in a patient with acute Stevens-Johnson syndrome

    Science.gov (United States)

    Ciralsky, Jessica B; Sippel, Kimberly C

    2013-01-01

    Background Stevens-Johnson syndrome is often associated with blinding ocular surface cicatricial sequelae. Recent reports have described markedly improved clinical outcomes with the application of amniotic membrane to the ocular surface during the acute phase. Here we describe the clinical outcome of a patient with acute Stevens-Johnson syndrome and severe ocular surface involvement in whom the evolving medical condition and family consent resulted in amniotic membrane application to each eye at differing intervals from disease onset. Methods We undertook a retrospective chart review of a woman with Stevens-Johnson syndrome who presented within hours of disease onset. She underwent application of amniotic membrane to the ocular surface of the left eye during the hyperacute phase (Stevens-Johnson syndrome proved superior. Application of amniotic membrane as soon as possible after disease onset, preferably in the hyperacute phase, appears to result in a significantly better clinical outcome than application later in the disease course. PMID:23754867

  13. [Biomechanic and biological activity assessment of concavity-convex amniotic membrane].

    Science.gov (United States)

    Shao, Yi; Yu, Yao; Tan, Gang; Peng, Juan; Zhou, Qiong; Pei, Chonggang; Dong, Wenjia; Gao, Guiping

    2012-12-01

    This paper conducted research on biomechanical characteristics and biological activity of concavity-convex amniotic membrane (CCAM) and discussed its superiority as ocular surface repair material. Folding and compression with vacuum of fresh amniotic membrane were used to prepare CCAM. After cutting the striga of CCAM, sixteen CCAM tissue section were chosen at random to test their tensile strength using electronic universal testing machine. The bilayer amniotic membrane (BAM), the double-deck amniotic membrane (DAM) and the monolayer amniotic membrane (MAM) were as controls. The test parameters included yield strength, tensile strength, elongation at break, elastic modulus and so on. The cytokines of fresh amniotic membrane (FAM), MAM and CCAM were analyzed by radioimmunoassay method. The CCAM was obviously thicker than MAM and DAM. After 15 min in PBS, the CCAM tissue can recover the normal shape. The tensile strength and the elongation at break of CCAM were higher than those of the MAM and the DAM (P biomechanical properties than the MAM and the DAM, showing the superiority as ocular surface repair material.

  14. Effect of Amniotic Membrane Combined with Ciprofloxacin in Curing the Primary Stages of Pseudomonal Keratitis

    Directory of Open Access Journals (Sweden)

    Mohammad Kazem Sharifi Yazdi

    2012-03-01

    Full Text Available Background: Keratitis caused by Pseudomonas aeruginosa is often resulted in severe corneal ulcers and perforation, which leads to losses of vision. Human amniotic membrane (HAM forms the inner wall of the membranous sac which surrounds and protects the embryo during gestation. The purpose of this study was to evaluate the effectiveness of the amniotic membrane's healing in rabbits with pseudomonas keratitis.Methods: In total 14 rabbits divided in 2 groups of: 1 as Control and 2 as experimental amniotic membrane combined with ciprofloxacin. A 0.05 ml suspension of Pseudomonas aeruginosa ATCC 27853 was injected into rabbit’s corneal stroma, with no interference in control group. In the second group, the amniotic membrane in pieces of 1.5 × 1.5 cm transplanted to the entire corneal surface by eight interrupted 10.0 nylon sutures. In the first day ciprofloxacin drop was injected to the second group every 30 minutes and through second to seventh days every 2 hours. The results of perforation in cornea and the amount of infiltration were registered.Results: The results showed that amniotic membrane transplantation (AMT + ciprofloxacin group had 0% perforation and the control group 85.6%. Average infiltrations were 5 mm in AMT + ciprofloxacin groups and 23.75 mm in control.Conclusion: The use of amniotic membrane with ciprofloxacin was effective in prevention of cornea perforation and controlling the process of pseudomonal keratitis remission. The improvement of inflammation rapidly happened in ciprofloxacin + AMT group.

  15. Evolution and homology of the astragalus in early amniotes: new fossils, new perspectives.

    Science.gov (United States)

    O'Keefe, F Robin; Sidor, Christian A; Larsson, Hans C E; Maga, Abdoudaye; Ide, Oumarou

    2006-04-01

    The reorganization of the ankle in basal amniotes has long been considered a key innovation allowing the evolution of more terrestrial and cursorial behavior. Understanding how this key innovation arose is a complex problem that largely concerns the homologizing of the amniote astragalus with the various ossifications in the anamniote tarsus. Over the last century, several hypotheses have been advanced homologizing the amniote astragalus with the many ossifications in the ankle of amphibian-grade tetrapods. There is an emerging consensus that the amniote astragalus is a complex structure emerging via the co-ossification of several originally separate elements, but the identities of these elements remain unclear. Here we present new fossil evidence bearing on this contentious question. A poorly ossified, juvenile astragalus of the large captorhinid Moradisaurus grandis shows clear evidence of four ossification centers, rather than of three centers or one center as posited in previous models of astragalus homology. Comparative material of the captorhinid Captorhinikos chozaensis is also interpretable as demonstrating four ossification centers. A new, four-center model for the homology of the amniote astragalus is advanced, and is discussed in the context of the phylogeny of the Captorhinidae in an attempt to identify the developmental transitions responsible for the observed pattern of ossification within this clade. Lastly, the broader implications for amniote phylogeny are discussed, concluding that the neomorphic pattern of astragalus ossification seen in all extant reptiles (including turtles) arose within the clade Diapsida.

  16. Amniotic band syndrome and/or limb body wall complex: split or lump

    Directory of Open Access Journals (Sweden)

    Ashutosh Halder

    2010-02-01

    Full Text Available Ashutosh HalderDepartment of Reproductive Biology, All India Institute of Medical Sciences, New Delhi, IndiaAbstract: Six cases of amniotic band syndrome/limb body wall complex were studied in respect to clinicopathologic characteristics. The diagnosis was based on two out of three of the following manifestations: cranio facial clefts; limb body wall defects and amniotic band attachment. Four cases were stillborn and associated with internal defects, including central nervous system. Two cases had facial and limb defects and were live born (3–5 years old at examination. Phenotypic features of the stillborn cases were craniofacial clefting, thoracoabdominoschisis, amputation, ring constriction, amniotic band adhesion, placental adhesions, and internal malformations. Histology of bands revealed fibroconnective tissue as well as flattened epithelial cells together with neuroectodermal elements. Umbilical cord section revealed an abnormal number of vessels. When analyzing the observed data in relation to their etiology, it was found that amniotic disruption, vascular disruption or genetic disruption could explain the amniotic band syndrome/limb body wall complexes, alone or in combinations. A brief review of literature in search of pathogenesis is offered along with an etiopathogenetic model.Keywords: amniotic band syndrome, limb body wall complex and pathogenesis

  17. Corneal regeneration by induced human buccal mucosa cultivated on an amniotic membrane following alkaline injury.

    Science.gov (United States)

    Man, Rohaina Che; Yong, Then Kong; Hwei, Ng Min; Halim, Wan Haslina Wan Abdul; Zahidin, Aida Zairani Mohd; Ramli, Roszalina; Saim, Aminuddin Bin; Idrus, Ruszymah Binti Hj

    2017-01-01

    Various clinical disorders and injuries, such as chemical, thermal, or mechanical injuries, may lead to corneal loss that results in blindness. PURPOSE : The aims of this study were to differentiate human buccal mucosa (BMuc) into corneal epithelial-like cells, to fabricate engineered corneal tissue using buccal mucosal epithelial cells, and to reconstruct a damaged corneal epithelium in a nude rat model. BMuc were subjected to 10 d of induction factors to investigate the potential of cells to differentiate into corneal lineages. Corneal stem cell markers β1-integrin, C/EBPδ, ABCG2, p63, and CK3 were upregulated in the gene expression analysis in induced BMuc, whereas CK3 and p63 showed significant protein expression in induced BMuc compared to the uninduced cells. BMuc were then left to reach 80% confluency after differential trypsinization. The cells were harvested and cultivated on a commercially available untreated air-dried amniotic membrane (AM) in a Transwell system in induction medium. The corneal constructs were fabricated and then implanted into damaged rat corneas for up to 8 weeks. A significant improvement was detected in the treatment group at 8 weeks post-implantation, as revealed by slit lamp biomicroscopy analysis. The structure and thickness of the corneal layer were also analyzed using histological staining and time-domain optical coherence tomography scans and were found to resemble a native corneal layer. The protein expression for CK3 and p63 were continuously detected throughout the corneal epithelial layer in the corneal construct. In conclusion, human BMuc can be induced to express a corneal epithelial-like phenotype. The addition of BMuc improves corneal clarity, prevents vascularization, increases corneal thickness and stromal alignment, and appears to have no adverse effect on the host after implantation.

  18. Clinical evaluation of the accuracy of intraocular pressure measurement by Tono-Pen XL in eyes with amniotic membrane patching.

    Science.gov (United States)

    Onguchi, Tatsuya; Takano, Yoji; Dogru, Murat; Tsubota, Kazuo; Shimazaki, Jun

    2005-03-01

    To evaluate the influence of amniotic membrane on intraocular pressure (IOP) measurement with the Tono-Pen. Observational case series. Eight patients with partial limbal dysfunction who underwent penetrating keratoplasty were recruited. Amniotic membranes were patched just after keratoplasty. One week postoperatively, amniotic membranes were removed, and accuracy of IOP measurement with the Tono-Pen was evaluated with comparisons before and after the removal. No epithelial defects were detected after the removal of amniotic membrane. Mean IOPs with and without amniotic membranes were 16.6 +/- 3.0 mm Hg (mean +/- SD) and 16.0 +/- 3.7 mm Hg, respectively. The difference was not statistically significant. Assessment of IOP with the Tono-Pen was observed to be accurate even when used over an amniotic membranes patch in cases who underwent penetrating keratoplasty.

  19. 4-H NFPA Fluid Power Challenge

    OpenAIRE

    Bonnett, Erika D

    2016-01-01

    The 4-H NFPA Fluid Power Challenge partnered Purdue Polytechnic Institute and Indiana 4-H with the National Fluid Power Association and Center for Compact and Efficient Fluid Power to provide teams of Indiana youth in 6-8th grades with opportunity to learn about hydraulics, engineering design, and other STEM skills. This created an opportunity to give youth a learning experience with STEM through hands-on, experiential learning activities. Youth experienced a one day workshop in which they wo...

  20. Therapeutic Evaluation of Mesenchymal Stem Cells in Chronic Gut Inflammation

    Science.gov (United States)

    2017-11-01

    and amniotic fluid obtained from healthy pregnancies [81]. Neverthe- less, the development of a newborn’s microbiota begins following birth via the...mediated destruction of pancreatic b cells involves both CD4+ and CD8+ T cells.164,165 A variety of different rat and mouse models of T1D have been used over...migrate to pancreatic islets through the interaction between chemokine stromal cell-derived factor 1 and chemokine receptor CXCR-4. Severe insulitis and b