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Sample records for human amniotic fluid-derived

  1. Human amniotic fluid derived cells can competently substitute dermal fibroblasts in a tissue-engineered dermo-epidermal skin analog

    NARCIS (Netherlands)

    Hartmann-Fritsch, Fabienne; Hosper, Nynke; Luginbuehl, Joachim; Biedermann, Thomas; Reichmann, Ernst; Meuli, Martin

    Human amniotic fluid comprises cells with high differentiation capacity, thus representing a potential cell source for skin tissue engineering. In this experimental study, we investigated the ability of human amniotic fluid derived cells to substitute dermal fibroblasts and support epidermis

  2. 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

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

    African Journals Online (AJOL)

    CLEMENTINA

    2012-06-28

    Jun 28, 2012 ... As a potential alternative treatment for terminal liver diseases, amniotic fluid derived mesenchymal stem cells (AFMSCs) have many advantages over other stem cells: avoiding much ethical controversy and decrease in both quantity and differentiation potential with age. However, the therapeutic role of.

  4. Human amniotic fluid-derived stem cells expressing cytosine deaminase and thymidine kinase inhibits the growth of breast cancer cells in cellular and xenograft mouse models.

    Science.gov (United States)

    Kang, N-H; Hwang, K-A; Yi, B-R; Lee, H J; Jeung, E-B; Kim, S U; Choi, K-C

    2012-06-01

    As human amniotic fluid-derived stem cells (hAFSCs) are capable of multiple lineage differentiation, extensive self-renewal and tumor targeting, they may be valuable for clinical anticancer therapies. In this study, we used hAFSCs as vehicles for targeted delivery of therapeutic suicide genes to breast cancer cells. hAFSCs were engineered to produce AF2.CD-TK cells in order to express two suicide genes encoding bacterial cytosine deaminase (CD) and herpes simplex virus thymidine kinase (HSV-TK) that convert non-toxic prodrugs, 5-fluorocytosine (5-FC) and mono-phosphorylate ganciclovir (GCV-MP), into cytotoxic metabolites, 5-fluorouracil (5-FU) and triphosphate ganciclovir (GCV-TP), respectively. In cell viability test in vitro, AF2.CD-TK cells inhibited the growth of MDA-MB-231 human breast cancer cells in the presence of the 5-FC or GCV prodrugs, or a combination of these two reagents. When the mixture of 5-FC and GCV was treated together, an additive cytotoxic effect was observed in the cell viability. In animal experiments using female BALB/c nude mouse xenografts, which developed by injecting MDA-MB-231 cells, treatment with AF2.CD-TK cells in the presence of 5-FC and GCV significantly reduced tumor volume and weight to the same extent seen in the mice treated with 5-FU. Histopathological and fluorescent staining assays further showed that AF2.CD-TK cells were located exactly at the site of tumor formation. Furthermore, breast tissues treated with AF2.CD-TK cells and two prodrugs maintained their normal structures (for example, the epidermis and reticular layers) while breast tissue structures in 5-FU-treated mice were almost destroyed by the potent cytotoxicity of the drug. Taken together, these results indicate that AF2.CD-TK cells can serve as excellent vehicles in a novel therapeutic cell-based gene-directed prodrug system to selectively target breast malignancies.

  5. Pre-Clinical Efficacy and Safety Evaluation of Human Amniotic Fluid-Derived Stem Cell Injection in a Mouse Model of Urinary Incontinence

    OpenAIRE

    Choi, Jae Young; Chun, So Young; Kim, Bum Soo; Kim, Hyun Tae; Yoo, Eun Sang; Shon, Yun-Hee; Lim, Jeong Ok; Yun, Seok Joong; Song, Phil Hyun; Chung, Sung Kwang; Yoo, James J; Kwon, Tae Gyun

    2015-01-01

    Purpose Stem cell-based therapies represent new promises for the treatment of urinary incontinence. This study was performed to assess optimized cell passage number, cell dose, therapeutic efficacy, feasibility, toxicity, and cell trafficking for the first step of the pre-clinical evaluation of human amniotic fluid stem cell (hAFSC) therapy in a urinary incontinence animal model. Materials and Methods The proper cell passage number was analyzed with hAFSCs at passages 4, 6, and 8 at week 2. T...

  6. Therapeutic outcomes of transplantation of amniotic fluid-derived stem cells in experimental ischemic stroke

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    Naoki eTajiri

    2014-08-01

    Full Text Available Accumulating preclinical evidence suggests the use of amnion as a source of stem cells for investigations of basic science concepts related to developmental cell biology, but also for stem cells’ therapeutic applications in treating human disorders. We previously reported isolation of viable rat amniotic fluid-derived stem (AFS cells. Subsequently, we recently reported the therapeutic benefits of intravenous transplantation of AFS cells in a rodent model of ischemic stroke. Parallel lines of investiagtions have provided safety and efficacy of stem cell therapy for treating stroke and other neurological disorders. This review article highlights characterization of AFS cells’ phenotype and their transplant-mediated functional effects, the need for investigations of mechanisms underlying AFS cells’ therapeutic benefits and discusses lab-to-clinic translational gating items in an effort to optimize the clinical application of cell transplantation for stroke.

  7. Pre-clinical efficacy and safety evaluation of human amniotic fluid-derived stem cell injection in a mouse model of urinary incontinence.

    Science.gov (United States)

    Choi, Jae Young; Chun, So Young; Kim, Bum Soo; Kim, Hyun Tae; Yoo, Eun Sang; Shon, Yun-Hee; Lim, Jeong Ok; Yun, Seok Joong; Song, Phil Hyun; Chung, Sung Kwang; Yoo, James J; Kwon, Tae Gyun

    2015-05-01

    Stem cell-based therapies represent new promises for the treatment of urinary incontinence. This study was performed to assess optimized cell passage number, cell dose, therapeutic efficacy, feasibility, toxicity, and cell trafficking for the first step of the pre-clinical evaluation of human amniotic fluid stem cell (hAFSC) therapy in a urinary incontinence animal model. The proper cell passage number was analyzed with hAFSCs at passages 4, 6, and 8 at week 2. The cell dose optimization included 1×10⁴, 1×10⁵, and 1×10⁶ cells at week 2. The in vivo cell toxicity was performed with 0.25×10⁶, 0.5×10⁶, and 1×10⁶ cells at weeks 2 and 4. Cell tracking was performed with 1×10⁶ cells at weeks 2 and 4. The selected optimal cell passage number was smaller than 6, and the optimal cell dose was 1×10⁶ for the mouse model. In our pre-clinical study, hAFSC-injected animals showed normal values for several parameters. Moreover, the injected cells were found to be non-toxic and non-tumorigenic. Furthermore, the injected hAFSCs were rarely identified by in vivo cell trafficking in the target organs at week 2. This study demonstrates for the first time the pre-clinical efficacy and safety of hAFSC injection in the urinary incontinence animal model and provides a basis for future clinical applications.

  8. Bioprinted Amniotic Fluid-Derived Stem Cells Accelerate Healing of Large Skin Wounds

    Science.gov (United States)

    Skardal, Aleksander; Mack, David; Kapetanovic, Edi; Atala, Anthony; Jackson, John D.; Yoo, James

    2012-01-01

    Stem cells obtained from amniotic fluid show high proliferative capacity in culture and multilineage differentiation potential. Because of the lack of significant immunogenicity and the ability of the amniotic fluid-derived stem (AFS) cells to modulate the inflammatory response, we investigated whether they could augment wound healing in a mouse model of skin regeneration. We used bioprinting technology to treat full-thickness skin wounds in nu/nu mice. AFS cells and bone marrow-derived mesenchymal stem cells (MSCs) were resuspended in fibrin-collagen gel and “printed” over the wound site. At days 0, 7, and 14, AFS cell- and MSC-driven wound closure and re-epithelialization were significantly greater than closure and re-epithelialization in wounds treated by fibrin-collagen gel only. Histological examination showed increased microvessel density and capillary diameters in the AFS cell-treated wounds compared with the MSC-treated wounds, whereas the skin treated only with gel showed the lowest amount of microvessels. However, tracking of fluorescently labeled AFS cells and MSCs revealed that the cells remained transiently and did not permanently integrate in the tissue. These observations suggest that the increased wound closure rates and angiogenesis may be due to delivery of secreted trophic factors, rather than direct cell-cell interactions. Accordingly, we performed proteomic analysis, which showed that AFS cells secreted a number of growth factors at concentrations higher than those of MSCs. In parallel, we showed that AFS cell-conditioned media induced endothelial cell migration in vitro. Taken together, our results indicate that bioprinting AFS cells could be an effective treatment for large-scale wounds and burns. PMID:23197691

  9. 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.

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

    Science.gov (United States)

    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.

  11. Predictive blood group genetics in hemolytic disease of the fetus and newborn: a 10-year review of a laboratory evaluation of amniotic fluid-derived DNA.

    Science.gov (United States)

    Seto, Eva; Fernandes, Bernard J; Wang, Chen; Denomme, Gregory A

    2007-11-01

    To identify the types of requests, the patterns of testing, and the error rates associated with the genetic identification of fetuses at risk for immune-mediated hemolytic disease. Retrospective review of the laboratory information system for all fetal blood group genotyping tests performed at Mount Sinai Hospital, Toronto, Canada from January 1997 to December 2006. Amniotic fluid-derived DNA, from 220 women (243 pregnancies), was tested for one or more antigens (279 tests) when the father was heterozygous for the inferred blood group antigen or was unavailable. The PCR amplification failure rate of amniotic fluid-derived DNA was 5.0%. When the father was considered hemizygous for RHD or was not available, the fetus was positive in 68.6% and 72.7% of cases, respectively. Two amniotic fluid-derived DNA KEL1 SSP-PCR results did not correlate with the result determined from cultured amniocyte DNA. Paternal RHD hemizygosity may be overestimated. Thus, we recommend that RHD zygosity be established by molecular analysis of the RHD breakpoint. Cultured amniocytes should be reserved for testing if the amniotic fluid-derived DNA is inconclusive due to PCR amplification failure. We use PCR-RFLP genotyping methods for RHCE*E/RHCE*e (Rh E/e), KEL1/KEL2 (K/k), FYA/FYB (Fy(a/b)), and JKA/JKB (Jk(a/b)).

  12. Amniotic fluid-derived mesenchymal stem cells prevent fibrosis and preserve renal function in a preclinical porcine model of kidney transplantation.

    Science.gov (United States)

    Baulier, Edouard; Favreau, Frederic; Le Corf, Amélie; Jayle, Christophe; Schneider, Fabrice; Goujon, Jean-Michel; Feraud, Olivier; Bennaceur-Griscelli, Annelise; Hauet, Thierry; Turhan, Ali G

    2014-07-01

    It is well known that ischemia/reperfusion injuries strongly affect the success of human organ transplantation. Development of interstitial fibrosis and tubular atrophy is the main deleterious phenomenon involved. Stem cells are a promising therapeutic tool already validated in various ischemic diseases. Amniotic fluid-derived mesenchymal stem cells (af-MSCs), a subpopulation of multipotent cells identified in amniotic fluid, are known to secrete growth factors and anti-inflammatory cytokines. In addition, these cells are easy to collect, present higher proliferation and self-renewal rates compared with other adult stem cells (ASCs), and are suitable for banking. Consequently, af-MSCs represent a promising source of stem cells for regenerative therapies in humans. To determine the efficiency and the safety of af-MSC infusion in a preclinical porcine model of renal autotransplantation, we injected autologous af-MSCs in the renal artery 6 days after transplantation. The af-MSC injection improved glomerular and tubular functions, leading to full renal function recovery and abrogated fibrosis development at 3 months. The strong proof of concept generated by this translational porcine model is a first step toward evaluation of af-MSC-based therapies in human kidney transplantation. ©AlphaMed Press.

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

    Science.gov (United States)

    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.

  14. In vitro fabrication of autologous living tissue-engineered vascular grafts based on prenatally harvested ovine amniotic fluid-derived stem cells.

    Science.gov (United States)

    Weber, Benedikt; Kehl, Debora; Bleul, Ulrich; Behr, Luc; Sammut, Sébastien; Frese, Laura; Ksiazek, Agnieszka; Achermann, Josef; Stranzinger, Gerald; Robert, Jérôme; Sanders, Bart; Sidler, Michele; Brokopp, Chad E; Proulx, Steven T; Frauenfelder, Thomas; Schoenauer, Roman; Emmert, Maximilian Y; Falk, Volkmar; Hoerstrup, Simon P

    2016-01-01

    Amniotic fluid cells (AFCs) have been proposed as a valuable source for tissue engineering and regenerative medicine. However, before clinical implementation, rigorous evaluation of this cell source in clinically relevant animal models accepted by regulatory authorities is indispensable. Today, the ovine model represents one of the most accepted preclinical animal models, in particular for cardiovascular applications. Here, we investigate the isolation and use of autologous ovine AFCs as cell source for cardiovascular tissue engineering applications. Fetal fluids were aspirated in vivo from pregnant ewes (n = 9) and from explanted uteri post mortem at different gestational ages (n = 91). Amniotic non-allantoic fluid nature was evaluated biochemically and in vivo samples were compared with post mortem reference samples. Isolated cells revealed an immunohistochemical phenotype similar to ovine bone marrow-derived mesenchymal stem cells (MSCs) and showed expression of stem cell factors described for embryonic stem cells, such as NANOG and STAT-3. Isolated ovine amniotic fluid-derived MSCs were screened for numeric chromosomal aberrations and successfully differentiated into several mesodermal phenotypes. Myofibroblastic ovine AFC lineages were then successfully used for the in vitro fabrication of small- and large-diameter tissue-engineered vascular grafts (n = 10) and cardiovascular patches (n = 34), laying the foundation for the use of this relevant pre-clinical in vivo assessment model for future amniotic fluid cell-based therapeutic applications. Copyright © 2013 John Wiley & Sons, Ltd.

  15. Human papillomavirus in amniotic fluid

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    Swan David C

    2006-09-01

    Full Text Available Abstract Background There is evidence to suggest that human papillomavirus (HPV can cross the placenta resulting in in-utero transmission. The goal of this study was to determine if HPV can be detected in amniotic fluid from women with intact amniotic membranes. Methods Residual amniotic fluid and cultured cell pellets from amniocentesis performed for prenatal diagnosis were used. PGMY09/11 L1 consensus primers and GP5+/GP6+ primers were used in a nested polymerase chain reaction assay for HPV. Results There were 146 paired samples from 142 women representing 139 singleton pregnancies, 2 twin pregnancies, and 1 triplet pregnancy. The women were 78% Caucasian, 5% African American, 14% Asian, and 2% Hispanic. The average age was 35.2 years with a range of 23–55 years. All samples were β-globin positive. HPV was not detected in any of the paired samples. Conclusion Given the age range, race, and ethnicity of the study population, one would anticipate some evidence of HPV if it could easily cross the placenta, but there was none.

  16. 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.

  17. 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.

  18. 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.

  19. Amniotic Fluid Derived Stem Cells with a Renal Progenitor Phenotype Inhibit Interstitial Fibrosis in Renal Ischemia and Reperfusion Injury in Rats.

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    Marina Gabriela Monteiro Carvalho Mori da Cunha

    Full Text Available Mesenchymal stem cells derived from human amniotic fluid (hAFSCs are a promising source for cellular therapy, especially for renal disorders, as a subpopulation is derived from the fetal urinary tract. The purpose of this study was to evaluate if hAFSCs with a renal progenitor phenotype demonstrate a nephroprotective effect in acute ischemia reperfusion (I/R model and prevent late stage fibrosis.A total of 45 male 12-wk-old Wistar rats were divided into three equal groups;: rats subjected to I/R injury and treated with Chang Medium, rats subjected to I/R injury and treated with hAFSCs and sham-operated animals. In the first part of this study, hAFSCs that highly expressed CD24, CD117, SIX2 and PAX2 were isolated and characterized. In the second part, renal I/R injury was induced in male rats and cellular treatment was performed 6 hours later via arterial injection. Functional and histological analyses were performed 24 hours, 48 hours and 2 months after treatment using serum creatinine, urine protein to creatinine ratio, inflammatory and regeneration markers and histomorphometric analysis of the kidney. Statistical analysis was performed by analysis of variance followed by the Tukey's test for multiple comparisons or by nonparametric Kruskal-Wallis followed by Dunn. Statistical significance level was defined as p <0.05.hAFSCs treatment resulted in significantly reduced serum creatinine level at 24 hours, less tubular necrosis, less hyaline cast formation, higher proliferation index, less inflammatory cell infiltration and less myofibroblasts at 48 h. The treated group had less fibrosis and proteinuria at 2 months after injury.hAFSCs contain a renal progenitor cell subpopulation that has a nephroprotective effect when delivered intra-arterially in rats with renal I/R injury, and reduces interstitial fibrosis on long term follow-up.

  20. Micelles of pulmonary surfactant in human amniotic fluid at term

    OpenAIRE

    Nishijima, Koji; SHUKUNAMI, Ken-ichi; Tsukahara, Hirokazu; ORISAKA, Makoto; Miura, Junichiro; KOTSUJI, Fumikazu

    2006-01-01

    Studies using in vitro analysis have shown that the interaction between pulmonary surfactant and vemix caseosa could explain the appearance of amniotic fluid turbidity. That phenomenon is interpreted based on the "roll-up" hypothesis. We tested the roll-up hypothesis by examining the presence of micelles of pulmonary surfactant in human amniotic fluid at term. Amniotic fluid samples were collected from each of six healthy pregnant women at term and at 16 wk of gestation. These samples were st...

  1. Critical assessment of antibacterial properties of human amniotic fluid.

    Science.gov (United States)

    Larsen, B; Davis, B; Charles, D

    1984-01-01

    A low molecular weight fraction of human amniotic fluid was found to be rapidly bactericidal for Escherichia coli ATCC 33908. Incubation in an atmosphere of CO2 but not N2 diminished the antibacterial effect suggesting that loss of CO2 and change in pH may be partly responsible for the observed antibacterial activity of the low molecular weight fraction of amniotic fluid. Incubation in pH 8.5 buffers did not duplicate the rapid killing of bacteria observed in the low molecular weight fraction of amniotic fluid, suggesting that elevated pH and an antibacterial factor from amniotic fluid act synergistically. The pH change in amniotic fluid which occurs in room atmosphere may account for some of the antibacterial effect which has been reported for amniotic fluid incubated aerobically.

  2. Micelles of pulmonary surfactant in human amniotic fluid at term.

    Science.gov (United States)

    Nishijima, Koji; Shukunami, Ken-Ichi; Tsukahara, Hirokazu; Orisaka, Makoto; Miura, Jun'Ichiro; Kotsuji, Fumikazu

    2006-08-01

    Studies using in vitro analysis have shown that the interaction between pulmonary surfactant and vernix caseosa could explain the appearance of amniotic fluid turbidity. That phenomenon is interpreted based on the "roll-up" hypothesis. We tested the roll-up hypothesis by examining the presence of micelles of pulmonary surfactant in human amniotic fluid at term. Amniotic fluid samples were collected from each of six healthy pregnant women at term and at 16 wk of gestation. These samples were stained negatively and analyzed using an electron microscope. Ultrastructures present in amniotic fluid were compared with the structure of micelles derived from suspended surfactant TA isolated from bovine lung. Surfactant TA formed spheroidal and rod-shaped micelles 10-70 nm in diameter above the critical micelle concentration. Identical micelle particles were described in human amniotic fluid at term. In addition, surfactant protein B was identified in the micelle fraction of amniotic fluid. However, no micelles were found in human amniotic fluid taken at 16 wk of gestation. Our results support the view that pulmonary surfactant could induce the detachment of vernix caseosa and increase the turbidity of the amniotic fluid.

  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. 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)

  5. [Intra-amniotic transmission of the human voice].

    Science.gov (United States)

    Querleu, D; Renard, X; Versyp, F; Paris-Delrue, L; Vervoort, P

    1988-01-01

    New acoustic measurements in amniotic environment permit to specify the conditions of transmission of human voices: voices emerge, incompletely covered by a low background noise, but however higher pitched near the placenta. The recognition of phonemes in utero is rather weak, approximately 30 per cent and appreciably identical for all voices and different mode of emission. Therefore this type of recognition is likely to play a minor role: the voices are evenly toneless by lack of high-pitch, and there is no obvious superiority of the intelligibility of direct maternal voice. The recognition of vowels by their second forming, as well as the emergence (demonstrated with special microphones) of impulse noises with very high-pitch components and of synthetic speech, confirm the possibility of transmission of frequencies exceeding 1,500 Hz to the intra-amniotic environment. On the contrary, melody recognition is excellent: probably major role of this factor. The loudness of the maternal voice transmitted to the uterus exceeds markedly that of outside voices, and this voice is certainly accessible to the fetus, most of the time. The demonstration of the transmission to the amniotic fluid of noises and voices enables to consider the possibility of perception.

  6. Human amniotic fluid stem cells have a potential to recover ovarian function in mice with chemotherapy-induced sterility.

    Science.gov (United States)

    Lai, Dongmei; Wang, Fangyuan; Chen, Yifei; Wang, Li; Wang, Yanlin; Cheng, Weiwei

    2013-09-04

    Human amniotic fluid cells (hAFCs) may differentiate into multiple cell lineages and thus have a great potential to become a donor cell source for regenerative medicine. The ability of hAFCs to differentiate into germ cell and oocyte-like cells has been previously documented. Herein we report the potential use of hAFCs to help restore follicles in clinical condition involving premature ovarian failure. Human amniotic fluid was obtained via amniocentesis, yielding a subpopulation of cloned hAFCs that was able to form embryoid bodies (EBs) and differentiate into three embryonic germ layers. Moreover, culture of EBs in medium containing human follicular fluid (HFF) or a germ cell maturation factor cocktail (FAC), expressed germ cells markers such as BLIMP1, STELLA, DAZL, VASA, STRA8, SCP3, SCP1, and GDF9. Furthermore, one cell line was grown from clone cells transfected with lentivirus-GFP and displaying morphological characteristics of mesenchymal cells, had the ability to restore ovarian morphology following cell injection into the ovaries of mice sterilized by intraperitoneal injection of cyclophosphamide and busulphan. Restored ovaries displayed many follicle-enclosed oocytes at all stages of development, but no oocytes or follicles were observed in sterilized mice whose ovaries had been injected with medium only (control). Notably, identification of GFP-labeled cells and immunostaining with anti-human antigen-specific antibodies demonstrated that grafted hAFCs survived and differentiated into granulosa cells which directed oocyte maturation. Furthermore, labeling of ovarian tissue for anti-Müllerian hormone expression, a functional marker of folliculogenesis, was strong in hAFCs-transplanted ovaries but inexistent in negative controls. These findings highlight the possibility of using human amniotic fluid-derived stem cells in regenerative medicine, in particular in the area of reproductive health.

  7. 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,

  8. Quantitative analysis of the toxicity of human amniotic fluid to cultured rat spinal cord.

    Science.gov (United States)

    Drewek, M J; Bruner, J P; Whetsell, W O; Tulipan, N

    1997-10-01

    It has been proposed that the myelodysplastic components of a myelomeningocele are secondarily damaged as the result of exposure to amniotic fluid, the so-called 'two-hit' hypothesis. The critical time at which this secondary insult might occur has not been clearly defined. The present study addresses this issue by quantitatively assessing the toxic effects of human amniotic fluid of various gestational ages upon organotypic cultures of rat spinal cord. Using an assay for lactate dehydrogenase efflux to evaluate toxicity in such spinal cord cultures, we found that the amniotic fluid became toxic at approximately 34 weeks' gestation. This toxic effect of amniotic fluid appears to emerge rather suddenly. Accordingly, it seems reasonable to suggest that prevention of exposure of vulnerable spinal cord tissue to this toxicity by surgical closure of a myelomeningocele defect prior to the emergence of toxicity in amniotic fluid may prevent injury to vulnerable myelodysplastic spinal cord tissue.

  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. Anxiolytic-Like Actions of Fatty Acids Identified in Human Amniotic Fluid

    Directory of Open Access Journals (Sweden)

    Rosa Isela García-Ríos

    2013-01-01

    Full Text Available Eight fatty acids (C12–C18 were previously identified in human amniotic fluid, colostrum, and milk in similar proportions but different amounts. Amniotic fluid is well known to be the natural environment for development in mammals. Interestingly, amniotic fluid and an artificial mixture of fatty acids contained in amniotic fluid produce similar anxiolytic-like actions in Wistar rats. We explored whether the lowest amount of fatty acids contained in amniotic fluid with respect to colostrum and milk produces such anxiolytic-like effects. Although a trend toward a dose-response effect was observed, only an amount of fatty acids that was similar to amniotic fluid fully mimicked the effect of diazepam (2 mg/kg, i.p. in the defensive burying test, an action devoid of effects on locomotor activity and motor coordination. Our results confirm that the amount of fatty acids contained in amniotic fluid is sufficient to produce anxiolytic-like effects, suggesting similar actions during intrauterine development.

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

    Directory of Open Access Journals (Sweden)

    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.

  12. Comparative analysis of the basement membrane composition of the human limbus epithelium and amniotic membrane epithelium.

    Science.gov (United States)

    Dietrich-Ntoukas, Tina; Hofmann-Rummelt, Carmen; Kruse, Friedrich E; Schlötzer-Schrehardt, Ursula

    2012-05-01

    Human amniotic membrane has been widely used as substrate for ex vivo expansion and transplantation of limbal epithelial cells. To further clarify its suitability as a surrogate niche for limbal stem cells and progenitor cells, we analyzed the composition of the amniotic epithelial basement membrane, with special focus on the expression of limbus-specific matrix components. Cryosections of corneoscleral specimens obtained from 10 human donor eyes and of 6 amniotic membrane specimens obtained at cesarean section were stained by indirect immunofluorescence using a broad panel of antibodies against basement membrane components. Both amniotic and limbal epithelial basement membranes showed positive immunoreactivity for collagen type IV α1, α2, α5, and α6 chains; collagens type VII, XV, XVI, XVII, and XVIII; laminin α3, β1, β2, β3, γ1, and γ2 chains; laminin-111 and laminin-332; nidogen-1 and nidogen-2; fibronectin; fibulin-2; fibrillin-2; perlecan; and agrin. Both types of basement membrane were negative for collagen type IV α3 and α4 chains, collagen type V, and laminin α4 chain. Limbal basement membrane components, which were not detected in amniotic membrane, included laminin α1, α2, α5, and γ3 chains; BM40/SPARC; tenascin-C; matrilin-2; endostatin; and collagen type XVIII. Despite extensive similarities in basement membrane composition between amniotic and corneolimbal epithelia, the lack of limbus-specific environmental factors argues against the potential of denuded amniotic membrane as a surrogate niche for limbal stem cells but supports its suitability as a substrate to promote the formation of a well-differentiated stratified corneal epithelial equivalent for tissue engineering strategies.

  13. Human amniotic epithelial cells express specific markers of nerve cells and migrate along the nerve fibers in the corpus callosum.

    Science.gov (United States)

    Wu, Zhiyuan; Hui, Guozhen; Lu, Yi; Liu, Tianjin; Huang, Qin; Guo, Lihe

    2012-01-05

    Human amniotic epithelial cells were isolated from a piece of fresh amnion. Using immunocytochemical methods, we investigated the expression of neuronal phenotypes (microtubule-associated protein-2, glial fibrillary acidic protein and nestin) in human amniotic epithelial cells. The conditioned medium of human amniotic epithelial cells promoted the growth and proliferation of rat glial cells cultured in vitro, and this effect was dose-dependent. Human amniotic epithelial cells were further transplanted into the corpus striatum of healthy adult rats and the grafted cells could integrate with the host and migrate 1-2 mm along the nerve fibers in corpus callosum. Our experimental findings indicate that human amniotic epithelial cells may be a new kind of seed cells for use in neurograft.

  14. Ultrastructural study of the neovagina following the utilization of human amniotic membrane for treatment of congenital absence of the vagina

    Directory of Open Access Journals (Sweden)

    L.F. Bleggi-Torres

    1997-07-01

    Full Text Available We present an ultrastructural study of the utilization of human amniotic membrane in the treatment of congenital absence of the vagina in 10 patients. All patients were surgically treated with application of an amniotic membrane graft using the modified McIndoe and Bannister technique. Sixty days after surgery, samples of the vaginal neo-epithelium were collected for transmission electron microscopy analysis. The ultrastructural findings consisted of a lining of mature squamous epithelium indicating the occurrence of metaplasia of the amniotic epithelium into the vaginal epithelium. The cells were arranged in layers as in the normal vaginal epithelium, i.e., superficial, intermediate and deep layers. There were desmosomes and cytoplasmic intermediate cytokeratin filaments, as well as some remnant features of the previous amniotic epithelium. These findings suggest that human amniotic membrane is able to complete metaplasia into squamous cells but the mechanism of this cellular transformation is unknown

  15. Effect of the Human Amniotic Membrane on Liver Regeneration in Rats

    Science.gov (United States)

    Sipahi, Mesut; Şahin, Sevinç; Arslan, Ergin; Börekci, Hasan; Metin, Bayram; Cantürk, Nuh Zafer

    2015-01-01

    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. PMID:26457000

  16. Effect of the Human Amniotic Membrane on Liver Regeneration in Rats

    Directory of Open Access Journals (Sweden)

    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.

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

    African Journals Online (AJOL)

    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 localization, and in vivo differentiation of AFMSC was confirmed by double-immunostaining ...

  18. Effects of Gamma Irradiation on Bacterial Microflora Associated with Human Amniotic Membrane

    Directory of Open Access Journals (Sweden)

    Fahmida Binte Atique

    2013-01-01

    Full Text Available Human amniotic membrane is considered a promising allograft material for the treatment of ocular surface reconstruction, burns, and other skin defects. In order to avoid the transmission of any diseases, grafts should be perfectly sterile. Twenty-five amniotic sacs were collected to determine the microbiological quality of human amniotic membrane, to analyze the radiation sensitivity pattern of the microorganism, and to detect the radiation decimal reduction dose (D10 values. All the samples were found to be contaminated, and the bioburden was ranged from 3.4×102 to 1.2×105 cfu/g. Initially, a total fifty bacterial isolates were characterized according to their cultural, morphological, and biochemical characteristics and then tested for the radiation sensitivity in an incremental series of radiation doses from 1 to 10 KGy. The results depict gradual decline in bioburden with incline of radiation doses. Staphylococcus spp. were the most frequently isolated bacterial contaminant in tissue samples (44%. The D10 values of the bacterial isolates were ranged from 0.6 to 1.27 KGy. Streptococcus spp. were found to be the highest radioresistant strain with the radiation sterilization dose (RSD of 11.4 KGy for a bioburden level of 1000. To compare the differences, D10 values were also calculated by graphical evaluations of the data with two of the representative isolates of each bacterial species which showed no significant variations. Findings of this study indicate that lower radiation dose is quite satisfactory for the sterilization of amniotic membrane grafts. Therefore, these findings would be helpful to predict the efficacy of radiation doses for the processing of amniotic membrane for various purposes.

  19. Effects of gamma irradiation on bacterial microflora associated with human amniotic membrane.

    Science.gov (United States)

    Binte Atique, Fahmida; Ahmed, Kazi Tahsin; Asaduzzaman, S M; Hasan, Kazi Nadim

    2013-01-01

    Human amniotic membrane is considered a promising allograft material for the treatment of ocular surface reconstruction, burns, and other skin defects. In order to avoid the transmission of any diseases, grafts should be perfectly sterile. Twenty-five amniotic sacs were collected to determine the microbiological quality of human amniotic membrane, to analyze the radiation sensitivity pattern of the microorganism, and to detect the radiation decimal reduction dose (D₁₀) values. All the samples were found to be contaminated, and the bioburden was ranged from 3.4 × 10² to 1.2 × 10⁵ cfu/g. Initially, a total fifty bacterial isolates were characterized according to their cultural, morphological, and biochemical characteristics and then tested for the radiation sensitivity in an incremental series of radiation doses from 1 to 10 KGy. The results depict gradual decline in bioburden with incline of radiation doses. Staphylococcus spp. were the most frequently isolated bacterial contaminant in tissue samples (44%). The D₁₀ values of the bacterial isolates were ranged from 0.6 to 1.27 KGy. Streptococcus spp. were found to be the highest radioresistant strain with the radiation sterilization dose (RSD) of 11.4 KGy for a bioburden level of 1000. To compare the differences, D₁₀ values were also calculated by graphical evaluations of the data with two of the representative isolates of each bacterial species which showed no significant variations. Findings of this study indicate that lower radiation dose is quite satisfactory for the sterilization of amniotic membrane grafts. Therefore, these findings would be helpful to predict the efficacy of radiation doses for the processing of amniotic membrane for various purposes.

  20. Amniotic fluid

    Science.gov (United States)

    ... or movements Too much amniotic fluid is called polyhydramnios . This condition can occur with multiple pregnancies (twins ... development of the fetus. Images Amniocentesis Amniotic fluid Polyhydramnios Amniotic fluid References Burton GJ, Sibley CP, Jauniaux ...

  1. Human amniotic fluid stem cells have a potential to recover ovarian function in mice with chemotherapy-induced sterility

    National Research Council Canada - National Science Library

    Dongmei Lai; Fangyuan Wang; Yifei Chen; Li Wang; Yanlin Wang; Weiwei Cheng

    2013-01-01

      Doc number: 34 Abstract Background: Human amniotic fluid cells (hAFCs) may differentiate into multiple cell lineages and thus have a great potential to become a donor cell source for regenerative medicine...

  2. Primary Adult Human Retinal Pigment Epithelial Cell Cultures on Human Amniotic Membranes

    Directory of Open Access Journals (Sweden)

    Singhal Shweta

    2005-01-01

    Full Text Available Purpose: Retinal pigment epithelial (RPE cells grow well on surfaces that provide an extracellular matrix. Our aim was to establish primary adult human RPE cell cultures that retain their epithelial morphology in vitro using human amniotic membrane (hAM as substrate. Materials and Methods: Human cadaver eyeballs (16 were obtained from the eye bank after corneal trephination. RPE cells were harvested by a mechanical dissection of the inner choroid surface (10, group 1 or by b enzymatic digestion using 0.25% Trypsin/0.02% EDTA (6, group 2. The cells were explanted onto de-epithelialized hAM, nourished using DMEM/HAMS F-12 media and monitored for growth under the phase contrast microscope. Cell cultures were characterised by whole mount studies and paraffin sections. Growth data in the two groups were compared using the students′ ′t′ test. Results: Eleven samples (68.75% showed positive cultures with small, hexagonal cells arising from around the explant which formed a confluent and progressively pigmented monolayer. Whole mounts showed closely placed polygonal cells with heavily pigmented cytoplasm and indistinct nuclei. The histologic sections showed monolayers of cuboidal epithelium with variable pigmentation within the cytoplasm. Growth was seen by day 6-23 (average 11.5 days in the mechanical group, significantly earlier ( P Conclusions: Primary adult human RPE cell cultures retain epithelial morphology in vitro when cultured on human amniotic membranes . Mechanical dissection of the inner choroid surface appears to be an effective method of isolating RPE cells and yields earlier growth in cultures as compared to isolation by enzymatic digestion

  3. A Preliminary Study of Human Amniotic Membrane as a Potential Chondrocyte Carrier

    Directory of Open Access Journals (Sweden)

    L Boo

    2009-11-01

    Full Text Available PURPOSE: To investigate the feasibility of using processed human amniotic membrane (HAM to support the attachment and proliferation of chondrocytes in vitro which in turn can be utilised as a cell delivery vehicle in tissue engineering applications. METHODS: Fresh HAM obtained from patients undergoing routine elective caesarean sections was harvested, processed and dried using either freeze drying (FD or air drying (AD methods prior to sterilisation by gamma irradiation. Isolated, processed and characterised rabbit autologous chondrocytes were seeded on processed HAM and cultured for up to three weeks. Cell attachment and proliferation were examined qualitatively using inverted brightfield microscopy. RESULTS: Processed HAM appeared to allow cell attachment when implanted with chondrocytes. Although cells seeded on AD and FD HAM did not appear to attach as strongly as those seeded on glycerol preserved intact human amniotic membrane, these cells to be proliferated in cell culture conditions. CONCLUSION: Preliminary results show that processed HAM promotes chondrocyte attachment and proliferation.

  4. Differentiation of mesenchymal stem cells from human amniotic fluid to vascular endothelial cells.

    Science.gov (United States)

    Tancharoen, Waleephan; Aungsuchawan, Sirinda; Pothacharoen, Peraphan; Markmee, Runchana; Narakornsak, Suteera; Kieodee, Junjira; Boonma, Nonglak; Tasuya, Witoon

    2017-03-01

    Endothelial dysfunction is a principle feature of vascular-related disease. Endothelial cells have been acquired for the purposes of the restoration of damaged tissue in therapeutic angiogenesis. However, their use is limited by expansion capacity and the small amount of cells that are obtained. Human amniotic fluid mesenchymal stem cells (hAF-MSCs) are considered an important source for vascular tissue engineering. In this study, hAF-MSCs were characterized and then induced in order to differentiate into the endothelial-like cells. Human amniotic fluid cells (hAFCs) were obtained from amniocentesis at the second trimester of gestation. The cells were characterized as mesenchymal stem cells by flow cytometry. The results showed that the cells were positive for mesenchymal stem cell markers CD44, CD73, CD90 and HLA-ABC, and negative for CD31, Amniotic fluid stem cells marker: CD117, anti-human fibroblasts, HLA-DR and hematopoietic differentiation markers CD34 and CD45. The hAF-MSCs were differentiated into endothelial cells under the induction of vascular endothelial growth factor (VEGF) and analyzed for the expression of the endothelial-specific markers and function. The expression of the endothelial-specific markers was determined by reverse transcriptase-quantitative PCR (RT-qPCR), while immunofluorescent analysis demonstrated that the induced hAF-MSCs expressed von Willebrand factor (vWF), vascular endothelial growth factor receptor 2 (VEGFR2), CD31 and endothelial nitric oxide synthase (eNOS). The network formation assay showed that the induced hAF-MSCs formed partial networks. All results indicated that hAF-MSCs have the potential to be differentiated into endothelial-like cells, while human amniotic fluid might be a suitable source of MSCs for vascularized tissue engineering. Copyright © 2016 Elsevier GmbH. All rights reserved.

  5. Human amniotic membrane as an intestinal patch for neomucosal growth in the rabbit model.

    Science.gov (United States)

    Barlas, M; Gökçora, H; Erekul, S; Dindar, H; Yücesan, S

    1992-05-01

    This experiment was carried out as a preliminary study, an attempt to grow new intestinal mucosa on human amniotic membrane in the terminal ileum in 37 rabbits. After ketamin sulfate anesthesia at laparatomy, 5-cm ileal defects were patched with human amniotic membrane (5 x 2 cm). These patched intestines were investigated on the first postoperative day and the 2nd, 5th, 10th, and 20th weeks corresponding to 4, 5, 5, 10, and 10 rabbits, respectively. Only three rabbits died in the early postoperative period. There was no evidence of intestinal obstruction or dilatation with barium meal. Microscopically, the neomucosa consisted of a thin layer of columnar epithelial cells at 2 weeks with more maturity of the villi and less irregularity and branching by 20 weeks. All patches were covered with neomucosa commencing at 2 weeks and covering the whole patch area by 20 weeks. This technique's advantages are the large size and the ease of the availability of the human amniotic membrane for neonates at risk without jeopardizing the neonates tissues. It is hoped that this method might be considered when neonatal material is scarce.

  6. The Effect of Reduced Bacterial Dilution on Human Amniotic Membrane Antibacterial Activity, in Vitro

    Directory of Open Access Journals (Sweden)

    Mohammad Mehdi Soltan-Dallal

    2013-05-01

    Full Text Available Background: The human amniotic membrane is the inner most layer of placenta and has antimicrobial effect, due to the presence of human beta-defensins and elafins. The purpose of this study is to investigate the effect of dilution reduction of 0.5 McFarland prepared from standard bacterial strains of Salmonella enterica BAA-708, Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853, Klebsiella pneumoniae ATCC7881, and Enterococcus faecalis ATCC29212 on antibacterial effect of human amniotic membranes in vitro.Materials and Methods: The amniotic membranes were obtained from the bank of organ transplantation in Imam Khomeini hospital, of women with elective cesarean section whose HIV, HBV, HCV and VDRL serological tests were negative. They were cut to 1.5×1.5 cm pieces. Then 0.5 McFarland suspensions of 1.5×108, 0.5×107 and 1.5×106 dilutions were prepared from bacteria which then were spread on Mueller Hinton medium agar and a piece of membrane was put in the center of each plate. After 24 hours incubation at 37ᵒC, the results were observed.Results: In 0.5 McFarland standard dilution an inhibition zone was created in three standard strains of Pseudomonas aeruginosa, Escherichia coli, and Salmonella enterica unlike the other two strains. There was no change in the above results with two other dilutions and inhibition zone of sensitive strains was not created.Conclusion: Dilution reduction of microbial strains does not affect the antibacterial impact of amniotic membrane and dilution reduction does not yield to a false positive response and the conversion of resistant to sensitive strains.

  7. DNA methylation patterns of imprinting centers for H19, SNRPN, and KCNQ1OT1 in single-cell clones of human amniotic fluid mesenchymal stem cell

    Directory of Open Access Journals (Sweden)

    Hsiu-Huei Peng

    2012-09-01

    Conclusion: In conclusion, human amniotic fluid mesenchymal stem cells contain a unique epigenetic signature during in vitro cell culture. H19 and KCNQ1OT1 possessed a substantial degree of hypermethylation status, and variable DNA methylation patterns of SNRPN was observed during in vitro cell culture of human amniotic fluid mesenchymal stem cells. Our results urge further understanding of epigenetic status of human amniotic fluid mesenchymal stem cells before it is applied in cell replacement therapy.

  8. Corneal haze induced by excimer laser photoablation in rabbits is reduced by preserved human amniotic membrane graft

    Science.gov (United States)

    Wang, Ming X.; Gray, Trevor; Prabhasawat, Pinnita; Ma, Xiong; Culbertson, William; Forster, Richard; Hanna, Khalil; Tseng, Scheffer C. G.

    1998-06-01

    We conducted a study to determine if preserved human amniotic membrane can reduce corneal haze induced by excimer laser photoablation. Excimer photoablation was performed bilaterally on 40 New Zealand white rabbits with a 6 mm ablation zone and 120 micrometer depth (PTK) using the VISX Star. One eye was randomly covered with a preserved human amniotic membrane and secured using four interrupted 10 - 0 nylon sutures; the other eye served as control. The amniotic membranes were removed at one week, and the corneal haze was graded with a slit-lamp biomicroscopy by three masked corneal specialists (WC, KH and RF) biweekly for the ensuing 12 weeks. Histology and in situ TUNEL staining (for fragmented DNA as an index for apoptosis) was performed at days 1, 3 and 7 and at 12 weeks. One week after excimer photoablation, the amniotic membrane-covered corneas showed more anterior stromal edema, which resolved at the second week. A consistent grading of organized reticular corneal haze was noted among the three masked observers. Such corneal haze peaked at the seventh week in both groups. The amniotic membrane-covered group showed statistically significant less corneal haze (0.50 plus or minus 0.15) than the control groups (1.25 plus or minus 0.35) (p less than 0.001). The amniotic membrane-covered corneas had less inflammatory response at days 1 and 3, showing nearly nil DNA fragmentation on keratocytes on the ablated anterior stromal and less stromal fibroblast activation. There is less altered epithelial cell morphology and less epithelial hyperplasia at 1 week in these amniotic membrane-treated eyes. We concluded from this study that amniotic membrane matrix is effective in reducing corneal haze induced by excimer photoablation in rabbits and may have clinical applications.

  9. Effect of Amniotic Membrane Suspension (AMS) and Amniotic Membrane Homogenate (AMH) on Human Corneal Epithelial Cell Viability, Migration and Proliferation In Vitro.

    Science.gov (United States)

    Wu, Ming-Feng; Stachon, Tanja; Langenbucher, Achim; Seitz, Berthold; Szentmáry, Nóra

    2017-03-01

    To analyze the effects of different concentrations of amniotic membrane suspension (AMS) or amniotic membrane homogenate (AMH) on human corneal epithelial cell (HCEC) viability, migration and proliferation. Amniotic membranes (AMs) of 13 placentas were prepared and stored at -80°C. For AMS preparation, following de-freezing, AM pieces were inserted in six-well plates and 5 ml Dulbecco's Modified Eagle's Medium (DMEM)/F12 (with 5% fetal bovine serum, FBS) per gram tissue was added for 96 h. After removal of the AM, the remaining supernatant was collected for experiments. For AMH preparation, following de-freezing, AMs were homogenized in liquid nitrogen and 5 ml DMEM/F12 (with 5% FBS) per gram tissue was added. Following centrifugation, the supernatant was collected for experiments. HCECs were expanded and incubated in DMEM/F12, 5% FBS supplemented by 15%, 30% or 100% AMS or 15% or 30% AMH. Viability was analyzed using Cell Proliferation Kit XTT, migration using wound healing assay and proliferation by the cell proliferation ELISA BrdU kit. HCEC viability remained unchanged using 15% or 30% AMS (p = 1.0 for both); however, it decreased significantly using 100% AMS (p migration increased significantly (p migration remained unchanged and 100% AMS inhibited HCEC migration (p migration, 15% and 30% AMS application seems to be the most appropriate method to support epithelial healing.

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

    Directory of Open Access Journals (Sweden)

    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

  11. Effects of Δ(9)-tetrahydrocannabinol (THC) on human amniotic epithelial cell proliferation and migration.

    Science.gov (United States)

    Yao, J L; He, Q Z; Liu, M; Chang, X W; Wu, J T; Duan, T; Wang, K

    2018-02-01

    The deleterious effects of cannabis consumption for fertility and pregnancy outcome are recognized for years. The main psychoactive molecule of cannabis, Δ(9)-tetrahydrocannabinol (THC) is able to cross the placenta barrier and cause alterations in fetal growth, low birth weight and preterm labor. However, the effects of THC on the human placenta amnion are still unknown. The distributions of CB1R and CB2R in human amnion tissues were observed by immunohistochemistry (IHC). Human amniotic epithelial cell proliferation and migration in response to THC treatment were measured by MTS and transwell assays, respectively. The PCR array was performed to study the key regulators involved in the cell migration. The protein levels of CB1R, CB2R in amnion tissues and MMP2, MMP9 in cells were detected by western blotting. Small interfering RNAs (siRNAs) were used to knockdown MMP2 and MMP9 in WISH cells. Our results indicated that both CB1R and CB2R primarily identified in the epithelial layer of human placental amnion tissue. The CB1R expression in the amnion tissue was higher in the preterm group than normal control. High-dose of THC (30uM, but not 20 and 10uM) significantly inhibited (p<0.01) human amniotic epithelial cell lines (WISH) proliferation. Meanwhile, THC at both 10uM and 20uM (p<0.05) significantly suppressed cells migration in both WISH and primary human amniotic epithelial cells. The PCR array data and siRNA experiments demonstrated that MMP2/9 were tightly involved in the regulation of THC-inhibited cell migration in WISH cells. These results suggested that THC inhibited the migration of human amniotic epithelial cell through the regulation of MMP2 and MMP9, which in turn altered the development of the amnion during the gestation and partially resulted in preterm labor and other adverse pregnancy outcomes. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Application of human amniotic mesenchymal cells as an allogeneic transplantation cell source in bone regenerative therapy

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    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.

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

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    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.

  14. 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.

  15. 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.

  16. 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.

  17. Using PRP and human amniotic fluid combination for osteogenesis in rabbit socket preservation

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    Amir Hossein Moradi

    2015-01-01

    Full Text Available Introduction: Platelet-rich plasma (PRP is used as an adjunct treatment during periodontal grafting surgery because of its capability of enhancing healing process. Amniotic fluid is a rich source of growth factors and hyaluronic acid (HA and a good point to study its properties of wound healing and bone formation. The aim of this study was to evaluate the osteogenic properties of a combination of amniotic fluid and PRP in rabbit′s dental socket preservation. Materials and Methods: The study population consisted of 24 healthy male laboratory rabbits (average weight 3,125 ± 185 gr that were randomly allocated into four groups. PRP for the first group, human amniotic fluid (HAF for the second group, a combination of PRP and HAF (PRHA for the third group was used. In the fourth (control group, no biomaterial was used. In each group, half of the rabbits were sacrificed at 4 weeks following surgery and the rest were sacrificed after 8 weeks. Histological analysis of biopsies of the sockets was performed using hematoxylin and eosin (H&E staining. Data were analyzed using Statistical Package for the Social Sciences (SPSS software (version 16 and P-value <0.05 was considered significance. Results: All three experimental groups showed positive effect on bone formation in terms of area of trabecular bone and number of osteocytes and also vessel formation. Socket preservation using HAF and PRHA showed the highest impact on bone formation. Socket preservation using HAF also had the highest impact on vessel formation. Conclusion: PRHA and HAF appear to be useful for enhancing bone formation. Since there was no difference between HAF and PRHA, it seems beneficial to use HAF due to its simplicity of application.

  18. Biological Activity Alterations of Human Amniotic Membrane Pre and Post Irradiation Tissue Banking.

    Science.gov (United States)

    Nemr, Waleed; Bashandy, A S; Araby, Eman; Khamiss, O

    Innate immunity of Human Amniotic Membrane (HAM) and its highly active secretome that rich with various types of growth factors and anti-inflammatory substances proposed it as a promising material for many medical studies and applications. This study evaluate the biological activity of cultivated HAM pre and post tissue banking process in which freeze-dried HAM was sterilized by 25 KGray (kGy) dose of γ radiation. The HAM's antimicrobial activity, viability, growth of isolated human amniotic epithelial cells (HAECs), hematopoietic stimulation of co-cultivated murine bone marrow cells (mammalian model), scaffold efficiency for fish brain building up (non-mammalian model) and self re-epithelialization after trypsin denuding treatment were examined as supposed biological activity features. Native HAM revealed viability indications and was active to kill all tested microorganisms; 6 bacterial species (3 Gram-positive and 3 Gram-negative) and Candida albicans as a pathogenic fungus. Also, HAM activity promoted colony formation of murine hematopoietic cells, Tilapia nilotica brain fragment building-up and self re-epithelialization after trypsin treatment. In contrary, radiation-based tissue banking of HAM caused HAM cellular death and consequently lacked almost all of examined biological activity features. Viable HAM was featured with biological activity than fixed HAM prepared by irradiation tissue banking.

  19. Management of temporomandibular joint degenerative disorders with human amniotic membrane: Hypothesis of action.

    Science.gov (United States)

    Guarda-Nardini, Luca; Trojan, Diletta; Paolin, Adolfo; Manfredini, Daniele

    2017-07-01

    Approaches providing the positioning of human amniotic membrane (HAM) within the intra-articular space of arthritic TMJs have never been investigated. This contrasts with the increasing amount of evidence suggesting the potential positive effects of HAM on a number of surgical conditions, even included the interpositional arthroplasty for TMJ ankylosis. Thus, the possible usefulness of HAM to restore joint functions in severely damaged TMJs could be hypothesized. Based on these premises, the clinical research question "Is human amniotic membrane positioning effective to reduce symptoms and restore jaw function in patients with severe inflammatory-degenerative disorders of the temporomandibular joint?" has been addressed by performing a systematic review of the literature. Out of potential 11988 and 8883 citations in the PubMed and Scopus databases, respectively, only five were of possible interest for inclusion in the review, but none of them addressed specifically the clinical research question. Thus, the hypothetical background for usefulness was discussed. The benefits of HAM positioning in TMJs with severe inflammatory-degenerative disorders could be related with its anti-inflammatory and anti-microbial and analgesic properties as well as its low immunogenicity. Studies in which HAM is positioned within the joint space of patients with severe TMJ degeneration, either as a disc-replacing film during major surgeries for discectomy and arthroplasty or as an injectable solution that can be needle-inserted after an arthrocentesis procedure, should be designed to test the hypothesis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Evaluation of visual acuity and color vision in normal human eyes with a sutureless temporary amniotic membrane patch.

    Science.gov (United States)

    Ijiri, Shigeyuki; Kobayashi, Akira; Sugiyama, Kazuhisa; Tseng, Scheffer C G

    2007-12-01

    To evaluate how sutureless amniotic membrane patches may affect visual functions in normal human eyes. Prospective intervention study. Ten sets of sutureless amniotic membrane patch manufactured as PROKERA were inserted in one eye of six normal patients. Four sets (one each) were inserted in four patients, while six sets (three each) were inserted in two patients. Uncorrected distant and near visual acuities, color vision, amniotic membrane thickness measured by pachymetry, and total symptom scores were compared before and after insertion. Within 30 minutes after insertion, mean distant visual acuities decreased from -0.22 +/- 0.06 to 0.92 +/- 0.45 logarithmic minimum angle of resolution (logMAR). Among 10 sets of PROKERA inserted, the largest optotype (1.0 logMAR) of the near vision chart could not be recognized in five, but color vision evaluated by Panel D-15 was still preserved in all. Total symptom scores increased to 47.8 +/- 9.1 points (maximum, 100 points). Among symptoms, total scores for foreign body sensation (17.8 +/- 3.6) and blurred vision (17.8 +/- 4.4) were high. Loss of distant visual acuity and increases of symptom scores were not correlated with amniotic membrane thickness, of which the mean was 67.6 +/- 25.2 mum. However, amniotic membrane that was less opaque tended to provide relatively good visual acuities. Because of the relative non-transparency of sutureless amniotic membrane patches in PROKERA, distant and near visual acuities decreased in normal human eyes. The foreign body sensation noted after insertion is primarily derived from the rigid supporting skirt.

  1. Atomized human amniotic mesenchymal stromal cells for direct delivery to the airway for treatment of lung injury

    NARCIS (Netherlands)

    Kim, Sally Yunsun; Burgess, Janette K.; Wang, Yiwei; Kable, Eleanor P. W.; Weiss, Daniel J.; Chan, Hak-Kim; Chrzanowski, Wojciech

    2016-01-01

    Background: Current treatment regimens for inhalation injury are mainly supportive and rely on self-regeneration processes for recovery. Cell therapy with mesenchymal stromal cells (MSCs) is increasingly being investigated for the treatment of inhalation injury. Human amniotic MSCs (hAMSCs) were

  2. Histological evaluation of human amniotic membrane graft on oral keratinizing mucosa in the rabbit model (A pilot study

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    Samandari Najafabadi MH

    2007-06-01

    Full Text Available Background and Aim: One of the complications following major oral surgeries is mucosal defects and delayed healing process. Up to now, various mucocutaneous grafts have been used in this field and recently, amniotic membrane has been proposed as a biological dressing in dermatologic, ophthalmologic and otolaryngologic practices. The purpose of this pilot study was to evaluate the healing process following human amniotic membrane graft on oral keratinized mucosa of rabbit.Materials and Methods: In this experimental animal study, two surgical mucosal defects with the same size were made in palatal mucosa of 10 rabbits with the same weight, gender and race and a graft of human amniotic membrane was used on one of the defects. On the 7th, 14th and 28th postoperative days, surgical biopsies were randomly obtained from grafted and ungrafted regions of 3, 4 and 3 rabbits, respectively and submitted for microscopic study. Results: According to the results, grafted regions showed more surface epithelialization and thicker newly formed epithelium. Also inflammatory cells infiltration was less in these areas. In all cases, there was a remarkable cartilage formation in the connective tissue of the recipient sites.Conclusion: The results of this study suggest that the use of amniotic membrane graft in oral surgery could be effective in healing process. Additional studies should be done using animal and human models with more samples. Furthermore, the formation of cartilage in the grafted sites and its possible potential in reconstruction of bone defects, needs to be studied.

  3. The development of a radioimmunoassay for reverse triiodothyronine sulfate in human serum and amniotic fluid

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Sing-Yung (Veterans Administration Medical Center, Long Beach, CA (United States)); Huang, Wen-Sheng; Chen, Wei-Lian (Tri-Service General Hospital, Taipei (Taiwan, Province of China)); Polk, D.; Reviczky, A.; Williams, J. III; Chopra, I.J.; Fisher, D.A. (Univ. of California, Los Angeles (United States))

    1993-06-01

    Sulfated iodothyronines including T[sub 4]-sulfate (T[sub 4]S) and T[sub 3]-sulfate (T[sub 3]S) have been identified in human serum and amniotic fluid. Little is know, however, about the existence of sulfate conjugation of reverse T[sub 3] (rT[sub 3]S) in man. In this report, the authors employed a novel, sensitive, and specific rT[sub 3]S RIA to address this question. The rabbit antiserum to rT[sub 3]S was highly specific; T[sub 4], T[sub 3], rT[sub 3], and 3,3'-T[sub 2] showed less than 0.002% cross-reaction with the antiserum. Only T[sub 4]S and T[sub 3]S cross-reacted significantly (0.3% and 0.01%, respectively); other analogs cross-reacted less than 0.0001%. The detection threshold of the RIA was 14 pmol/L (1.0 ng/dL). The mean serum rT[sub 3]S concentration (pmol/L) was 40 in euthyroid subjects. Values were similar in hypothyroid patients (38) and pregnant women (52) but significantly (P < 0.01) elevated to 176 in hyperthyroid patient, 74 in patients with nonthyroid illnesses, and 684 in cord sera of newborns. Serum rT[sub 3]S increased significantly in hyperthyroid patients 1 day after administration of 1 g sodium ipodate orally. Reverse T[sub 3]S was detected consistently in amniotic fluid at 14 to 22 weeks of gestation and showed a marked rise 1-3 weeks after intraamniotic administration of 500-1000 [mu]g T[sub 4]. The various data suggest that : (1) rT[sub 3]S is a normal component of human serum and amniotic fluid; (2) it is derived from metabolism of T[sub 4] or rT[sub 3]; (3) circulating rT[sub 3]S increases in hyperthyroidism and in circumstances where type I 5'-monodeiodinating activity is low, e.g. nonthyroid illnesses, fetal life, and after administration of ipodate. 20 refs., 4 figs.

  4. Human amniotic epithelial cell transplantation for the repair of injured brachial plexus nerve: evaluation of nerve viscoelastic properties

    Science.gov (United States)

    Jin, Hua; Yang, Qi; Ji, Feng; Zhang, Ya-jie; Zhao, Yan; Luo, Min

    2015-01-01

    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 C6 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 C6 brachial plexus injury site (1 × 106 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 C6 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. PMID:25883625

  5. 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.

  6. Exosomes derived from human amniotic epithelial cells accelerate wound healing and inhibit scar formation.

    Science.gov (United States)

    Zhao, Bin; Zhang, Yijie; Han, Shichao; Zhang, Wei; Zhou, Qin; Guan, Hao; Liu, Jiaqi; Shi, Jihong; Su, Linlin; Hu, Dahai

    2017-04-01

    Wound healing is a highly orchestrated physiological process consisting of a complex events, and scarless wound healing is highly desired for the development and application in clinical medicine. Recently, we have demonstrated that human amniotic epithelial cells (hAECs) promoted wound healing and inhibited scar formation through a paracrine mechanism. However, exosomes (Exo) are one of the most important paracrine factors. Whether exosomes derived from human amniotic epithelial cells (hAECs-Exo) have positive effects on scarless wound healing have not been reported yet. In this study, we examined the role of hAECs-Exo on wound healing in a rat model. We found that hAECs, which exhibit characteristics of both embryonic and mesenchymal stem cells, have the potential to differentiate into all three germ layers. hAECs-Exo ranged from 50 to 150 nm in diameter, and positive for exosomal markers CD9, CD63, CD81, Alix, TSG101 and HLA-G. Internalization of hAECs-Exo promoted the migration and proliferation of fibroblasts. Moreover, the deposition of extracellular matrix (ECM) were partly abolished by the treatment of high concentration of hAECs-Exo (100 μg/mL), which may be through stimulating the expression of matrix metalloproteinase-1 (MMP-1). In vivo animal experiments showed that hAECs-Exo improved the skin wound healing with well-organized collagen fibers. Taken together, These findings represent that hAECs-Exo can be used as a novel hope in cell-free therapy for scarless wound healing.

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

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    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.

  9. 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.

  10. 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.

  11. Antimicrobial and Antibiofilm Effects of Human Amniotic/Chorionic Membrane Extract on Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    Mukesh K. Yadav

    2017-10-01

    Full Text Available Background:Streptococcus pneumoniae colonize the human nasopharynx in the form of biofilms. The biofilms act as bacterial reservoirs and planktonic bacteria from these biofilms can migrate to other sterile anatomical sites to cause pneumonia, otitis media (OM, bacteremia and meningitis. Human amniotic membrane contains numerous growth factors and antimicrobial activity; however, these have not been studied in detail. In this study, we prepared amniotic membrane extract and chorionic membrane extract (AME/CME and evaluated their antibacterial and antibiofilm activities against S. pneumoniae using an in vitro biofilm model and in vivo OM rat model.Materials and Methods: The AME/CME were prepared and protein was quantified using DCTM (detergent compatible method. The minimum inhibitory concentrations were determined using broth dilution method, and the synergistic effect of AME/CME with Penicillin-streptomycin was detected checkerboard. The in vitro biofilm and in vivo colonization of S. pneumoniae were studied using microtiter plate assay and OM rat model, respectively. The AME/CME-treated biofilms were examined using scanning electron microscope and confocal microscopy. To examine the constituents of AME/CME, we determined the proteins and peptides of AME/CME using tandem mass tag-based quantitative mass spectrometry.Results: AME/CME treatment significantly (p < 0.05 inhibited S. pneumoniae growth in planktonic form and in biofilms. Combined application of AME/CME and Penicillin-streptomycin solution had a synergistic effect against S. pneumoniae. Biofilms grown with AME/CME were thin, scattered, and unorganized. AME/CME effectively eradicated pre-established pneumococci biofilms and has a bactericidal effect. AME treatment significantly (p < 0.05 reduced bacterial colonization in the rat middle ear. The proteomics analysis revealed that the AME/CME contains hydrolase, ribonuclease, protease, and other antimicrobial proteins and peptides

  12. Rapid screening and multi-toxin profile confirmation of tetrodotoxins and analogues in human body fluids derived from a puffer fish poisoning incident in New Caledonia.

    Science.gov (United States)

    Rambla-Alegre, Maria; Leonardo, Sandra; Barguil, Yann; Flores, Cintia; Caixach, Josep; Campbell, Katrina; Elliott, Christopher T; Maillaud, Claude; Boundy, Michael J; Harwood, D Tim; Campàs, Mònica; Diogène, Jorge

    2018-02-01

    In August 2014, a puffer fish poisoning incidence resulting in one fatality was reported in New Caledonia. Although tetrodotoxin (TTX) intoxication was established from the patients' signs and symptoms, the determination of TTX in the patient's urine, serum or plasma is essential to confirm the clinical diagnosis. To provide a simple cost-effective rapid screening tool for clinical analysis, a maleimide-based enzyme-linked immunosorbent assay (mELISA) adapted for the determination of TTX contents in human body fluids was assessed. The mELISA was applied to the analysis of urine samples from two patients and a response for the presence of TTX and/or structurally similar analogues was detected in all samples. The analysis by LC-MS/MS confirmed the presence of TTX but also TTX analogues (4-epiTTX, 4,9-anhydroTTX and 5,6,11-trideoxyTTX) in the urine. A change in the multi-toxin profile in the urine based on time following consumption was observed. LC-MS/MS analysis of serum and plasma samples also revealed the presence of TTX (32.9 ng/mL) and 5,6,11-trideoxyTTX (374.6 ng/mL) in the post-mortem plasma. The results provide for the first time the TTX multi-toxin profile of human samples from a puffer fish intoxication and clearly demonstrate the implication of TTX as the causative agent of the reported intoxication case. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. A Comparison of Culture Characteristics between Human Amniotic Mesenchymal Stem Cells and Dental Stem Cells.

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    Yusoff, Nurul Hidayat; Alshehadat, Saaid Ayesh; Azlina, Ahmad; Kannan, Thirumulu Ponnuraj; Hamid, Suzina Sheikh Abdul

    2015-04-01

    In the past decade, the field of stem cell biology is of major interest among researchers due to its broad therapeutic potential. Stem cells are a class of undifferentiated cells that are able to differentiate into specialised cell types. Stem cells can be classified into two main types: adult stem cells (adult tissues) and embryonic stem cells (embryos formed during the blastocyst phase of embryological development). This review will discuss two types of adult mesenchymal stem cells, dental stem cells and amniotic stem cells, with respect to their differentiation lineages, passage numbers and animal model studies. Amniotic stem cells have a greater number of differentiation lineages than dental stem cells. On the contrary, dental stem cells showed the highest number of passages compared to amniotic stem cells. For tissue regeneration based on animal studies, amniotic stem cells showed the shortest time to regenerate in comparison with dental stem cells.

  14. Isolation and Partial Characterization of Human Amniotic Epithelial Cells: The Effect of Trypsin

    Science.gov (United States)

    Tabatabaei, Meraj; Mosaffa, Nariman; Nikoo, Shohreh; Bozorgmehr, Mahmood; Ghods, Roya; Kazemnejad, Somaieh; Rezania, Simin; Keshavarzi, Bahareh; Arefi, Soheila; Ramezani-Tehrani, Fahimeh; Mirzadegan, Ebrahim; Zarnani, Amir-Hassan

    2014-01-01

    Background Despite the extensive information available in the literature, cell surface marker signature of human Amniotic Epithelial Cells (hAECs) remains controversial. The aim of the present study was to characterize immunophenotypic features, proliferative capacity and immunogenicity of hAECs. We also tested whether expression of some cell surface markers is influenced by the type of trypsin used for tissue digestion. Methods Single cell suspensions of amniotic membranes from four human placentas were isolated by enzymatic digestion and expression of CD9, CD10, CD29, CD34, CD38, CD44, CD45, CD73, CD105, CD133, HLA-I, HLA-DR, HLA-G, SSEA-4, STRO-1 and OCT-4 was then evaluated by flow cytometry. The differential impact of four trypsin types on the yield and expression of CD105 and HLA-I was also determined. The proliferative capacity of cultured hAECs was assessed and compared in the presence and absence of Epidermal Growth Factor (EGF). To test their immunogenicity, hAECs were injected into Balb/c mice and the reactivity of hyperimmunized sera was examined by immunofluorescence staining. Results Nearly all purified cells expressed mesenchymal markers, CD9, CD10, CD29, and CD73 and the embryonic marker, SSEA-4. A large proportion of the cells also expressed STRO-1 and OCT-4. The purified cells also expressed HLA-G and HLA-I. A very small proportion of hAECs expressed CD34, CD38, CD44, CD133 and HLA-DR. The type of trypsin used for enzymatic digestion affected both the percentage and expression of HLA-I and CD105. hAECs revealed substantial proliferative capacity only when cultured in the medium supplemented with EGF. These cells were shown to be capable of inducing high amounts of anti-donor antibodies. Conclusion Here we provided evidence that hAECs are immunogenic cells with high level of HLA-I expression. Furthermore, this work highlighted the impact of isolation procedure on the immunophenotype of hAEC. PMID:24523953

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

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    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.

  16. 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.

  17. [Procedural guidelines. Good practice procedures for acquisition and preparation of cryopreserved human amniotic membranes from donor placentas].

    Science.gov (United States)

    Hahn, A; Thanos, M; Reinhard, T; Seitz, B; Steuhl, K-P; Meller, D

    2010-11-01

    A cornea/tissue bank must have an organizational structure in which responsibility and authority to issue directives are clearly defined. It must also use a documented quality management system on the basis of good practice procedures which is maintained to the current standards. The personnel of a cornea/tissue bank must be present in sufficient numbers and be suitably qualified. A cornea/tissue bank must be in possession of appropriate facilities which are suitable for the main purpose of preparation of cryopreserved human amniotic membranes from donor placentas. All equipment must be designed and maintained corresponding to the intended purpose. Deviations from the stipulated quality and safety standards must give rise to documented investigations which include decisions on options for correctional and preventive measures. Acquisition of donors and tissue sampling must be strictly controlled and documented. This also applies to entry of donor tissue in the cornea/tissue bank. Cryopreserved human amniotic membranes can only be preserved from donors undergoing caesarean section and who did not present any known infection of the abdominal cavity or any systemic blood borne infection. Contamination of media used for cryopreservation of donor placenta must be ruled out at least once. Measures must be taken to keep the risk of contamination as low as possible. Cryopreserved human amniotic membranes from donor placentas can only be released if defined criteria are fulfilled. Any suspicion of severe undesired reactions and events for the recipient of an amniotic membrane transplant must be registered with the authorities. The activities of a cornea/tissue bank must maintain and adapt to the state-of-the-art with respect to scientific progress.

  18. Genotoxic effects of cadmium chloride on human amniotic fluid cells cultured in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Fogu, G. [Sassari Univ., Sassari (Italy). Dept. of Physiological, Biochemical and Cellular sciences; Sassari Univ., Sassari (Italy). Centre of Clinical Genetics; Congiu, A. M.; Sini, M. C.; Ladu, R. [Sassari Univ., Sassari (Italy). Dept. of Physiological, Biochemical and Cellular sciences; Campus, P. M.; Sanna, R.; Soro, G. [Sassari Univ., Sassari (Italy). Centre of Clinical Genetics

    2000-12-01

    In this study it has been reported the results of cytogenetic tests, namely a search for chromosome aberrations (CA) and sister chromatid exchanges (SCEs), performed on human amniotic fluid cells cultured and treated with Cadmium chloride. The cells from primary cultures were exposed to CdCl{sub 2} at 1 {mu}M and 10 {mu}M for 24 h. At the higher dose, no metaphases were scored and at the lower dose (1 {mu}M) no effects were evident on cell proliferation, and no chromosome aberrations were found. In the subsequent experiments were used cells from subcultures exposed to 1 {mu}M and 5 {mu}M CdCl{sub 2}. At the 5 {mu}M dose was evident the induction of chromatid breaks, while the frequency of sister chromatid exchanges shows a small increase, not statistically significant at the dose of 1 {mu}M. In this study it was positively demonstrated that amniotic fluid cells grown in vitro are reliable for testing various mutagenic or teratogenic substances. With regard to cadmium treatment results, it is evident a clastogenic effect of cadmium chloride but not a significant induction of SCEs. [Italian] In questo studio abbiamo riportato i risultati dei tests citogenici di valutazione degli scambi fra cromatidi fratelli (SCEs) e di induzione di aberrazioni cromosomiche (CA) condotti su colture cellulari di liquido amniotico umano, trattate con cloruro di cadmio. Le cellule delle colture primarie venivano esposte al CdCl{sub 2} a concentrazioni di 1 {mu}M 10 {mu}M per 24 h. Alla dose piu' alta (10 {mu}M) non sono state osservate metafasi, mentre alla dose piu' bassa (1 {mu}M) non sono stati evidenziati ne' effetti sulla proliferazione cellulare ne' CA. Nei successivi esperimenti il CdC{sub 2} e' stato testato su subcolture cellulari alle dosi di 1 {mu}M e 5 {mu}M. Alla dose di 5 {mu}M sono state evidenziate rotture cromatidiche, mentre la frequenza di SCE alla dose di 1 {mu}M ha mostrato un piccolo incremento, statisticamente non significativo. In questo

  19. Place of human amniotic membrane immunoblotting in the diagnosis of autoimmune bullous dermatoses.

    Science.gov (United States)

    Grootenboer-Mignot, S; Descamps, V; Picard-Dahan, C; Nicaise-Roland, P; Prost-Squarcioni, C; Leroux-Villet, C; Champagnat, C; Delaval, A; Aucouturier, F; Crickx, B; Chollet-Martin, S

    2010-04-01

    Fine analysis of antiskin autoantibodies can contribute to the differential diagnosis of autoimmune bullous dermatoses. To develop a high-performance immunoblotting method using human amniotic membrane as the antigen source, and to compare it with current laboratory methods. Sera from 113 patients were tested by immunoblotting (IB), rat and monkey oesophagus and salt-split skin indirect immunofluorescence (IIF), and enzyme-linked immunosorbent assay (ELISA) quantification of anti-BP180-NC16a and anti-BP230, or antidesmoglein (Dsg) 1 and 3 antibodies. There were 56 cases of bullous pemphigoid (BP), 22 cases of mucous membrane pemphigoid (MMP), eight cases of epidermolysis bullosa acquisita (EBA), two cases of bullous systemic lupus erythematosus (BSLE), 17 cases of pemphigus vulgaris (PV), and four cases each of pemphigus foliaceus (PF) and paraneoplastic pemphigus (PNP). In BP, the three methods had similar sensitivity (84-89%) for both anti-BP180-NC16a and anti-BP230 antibody detection. In MMP, autoantibodies (mainly directed against BP180 or laminin 332 subunits) were detected in 77% of patients by IB, compared with only 9% by IIF on rat and monkey oesophagus and 36% on salt-split skin, and 14% by anti-BP180-NC16a and anti-BP230 ELISA. In patients with pemphigus, ELISA had 92% sensitivity for anti-Dsg1 and 3, but IB and rat bladder IIF were necessary to confirm PNP by revealing specific and rare patterns (antidesmoplakin I/II, antienvoplakin and antiperiplakin antibodies). IB also revealed anticollagen VII antibodies in 60% of patients with EBA and BSLE, and antibodies to BP180, BP230 and Dsg3 in a few patients who were negative using the other two techniques. Amniotic membrane immunoblotting is an interesting diagnostic tool for bullous diseases, as the entire panel of autoantibodies can be detected with a single extract. This method improves the identification of complex and heterogeneous autoimmune processes in conjunction with IIF and ELISA, and is

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

    Directory of Open Access Journals (Sweden)

    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

  1. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Blount, Benjamin C. [Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341 (United States)]. E-mail: bblount@cdc.gov; Valentin-Blasini, Liza [Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341 (United States)

    2006-05-10

    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{sup 18}O{sub 4} {sup -}, S{sup 13}CN{sup -} and {sup 15}NO{sub 3} {sup -} 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 {mu}g/L), thiocyanate (<10-5860, 89 {mu}g/L), nitrate (650-8900, 1620 {mu}g/L) and iodide (1.7-170, 8.1 {mu}g/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.

  3. Technique of cultivating limbal derived corneal epithelium on human amniotic membrane for clinical transplantation

    Directory of Open Access Journals (Sweden)

    Fatima A

    2006-01-01

    Full Text Available Background : The technique of transplantation of cultivated limbal epithelium rather than direct limbal tissue isa novel method of "cell therapy" involved in reconstructing the ocular surface in severe limbal stem celldeficiency [LSCD], caused by chemical burns. Aim : To describe a simple feeder-cell free technique of cultivating limbal epithelium on human amniotic membrane[HAM]. Materials and Methods : The limbal tissues (2 mm were harvested from patients with LSCD. These tissueswere proliferated in vitro on HAM supplemented by human corneal epithelial cell medium and autologousserum. Cultures covering more ?50% area of 2.5x5 cm HAM were considered adequate for clinical use. Thecultured epithelium was characterized by histopathology and immunophenotyping.Results: A total of 542 cultures out of 250 limbal tissues were cultivated in the laboratory from January 2001through July 2005. The culture explants showed that clusters of cells emerging from the edge of the explantsin one-three days formed a complete monolayer within 10-14 days. In 86% of cultures (464 of 542, thegrowth was observed within one-two days. Successful explant cultures were observed in 98.5% (534 of 542cultures with 91% explant cultures showing an area of ?6.25 cm2 (6.25 - 12.5 cm2 range. The cultivatedepithelium was terminated between 10-14 days for clinical transplantation. The problems encountered wereinadequate growth (2 of 542 and contamination (2 of 542. Conclusions : We demonstrate a simple technique of generating a sheet of corneal epithelium from a limbalbiopsy. This new technique could pave the way for a novel form of cell therapy.

  4. 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.

  5. Differentiation of mesenchymal stem cells from human amniotic fluid to cardiomyocyte‑like cells.

    Science.gov (United States)

    Markmee, Runchana; Aungsuchawan, Sirinda; Narakornsak, Suteera; Tancharoen, Waleephan; Bumrungkit, Kanokkarn; Pangchaidee, Nataporn; Pothacharoen, Peraphan; Puaninta, Chaniporn

    2017-11-01

    Ischemic heart disease (IHD) is a major factor influencing worldwide mortality rates. Furthermore, IHD has become a significant health problem among the Thai population. Stem cell therapy using mesenchymal stem cells (MSCs) is an alternative therapeutic method that has been applied to improve the quality of life of patients. Amniotic fluid (AF) contains a heterogeneous cell population, including MSCs, which are multipotent stem cells that have the capability to differentiate into mesenchymal lineages. The purpose of the present study was to evaluate the MSC characteristics of human (h)AF and determine its potency regarding cardiogenic differentiation. MSC characterization following flow cytometric analysis revealed that the cells expressed MSC markers, cluster of differentiation (CD)44, CD90, human leukocyte antigen‑ABC and CD73. The results of the alamar blue assay demonstrated that cell proliferation rate continuously increased from the early cultivation phase up to 5‑fold during days 1 to 5 of cell culturing. The highest rate of cell proliferation was observed on day 17 with a 30‑fold increase compared with that on day 1. During the cardiogenic induction stage, morphological changes were observed between day 0 and day 21, and it was revealed that the hAF derived‑MSCs in the cardiogenic‑induced group exhibited myotube‑like morphology after 7 days of cell culturing. Following cardiogenic induction, immunohistochemistry staining was performed on day 21, and reverse transcription‑quantitative polymerase chain reaction on day 7 and 21. These steps were performed to detect the protein and gene expression levels of cardiac specific proteins (GATA4, cardiac troponin T, Nkx2.5 and Connexin43). The results of the present study indicated that hAF‑MSCs possess the potential to differentiate into cardiomyocyte‑like cells. Thus, it was concluded that hAF may be a suitable source of MSCs for stem cell therapy and tissue engineering.

  6. In vitro differentiation into insulin-producing β-cells of stem cells isolated from human amniotic fluid and dental pulp.

    Science.gov (United States)

    Carnevale, Gianluca; Riccio, Massimo; Pisciotta, Alessandra; Beretti, Francesca; Maraldi, Tullia; Zavatti, Manuela; Cavallini, Gian Maria; La Sala, Giovanni Battista; Ferrari, Adriano; De Pol, Anto

    2013-08-01

    To investigate the ability of human amniotic fluid stem cells and human dental pulp stem cells to differentiate into insulin-producing cells. Human amniotic fluid stem cells and human dental pulp stem cells were induced to differentiate into pancreatic β-cells by a multistep protocol. Islet-like structures were assessed in differentiated human amniotic fluid stem cells and human dental pulp stem cells after 21 days of culture by dithizone staining. Pancreatic and duodenal homebox-1, insulin and Glut-2 expression were detected by immunofluorescence and confocal microscopy. Insulin secreted from differentiated cells was tested with SELDI-TOF MS and by enzyme-linked immunosorbent assay. Human amniotic fluid stem cells and human dental pulp stem cells, after 7 days of differentiation started to form islet-like structures that became evident after 14 days of induction. SELDI-TOF MS analysis, revealed the presence of insulin in the media of differentiated cells at day 14, further confirmed by enzyme-linked immunosorbent assay after 7, 14 and 21 days. Both stem cell types expressed, after differentiation, pancreatic and duodenal homebox-1, insulin and Glut-2 and were positively stained by dithizone. Either the cytosol to nucleus translocation of pancreatic and duodenal homebox-1, either the expression of insulin, are regulated by glucose concentration changes. Day 21 islet-like structures derived from both human amniotic fluid stem cells and human dental pulp stem cell release insulin in a glucose-dependent manner. The present study demonstrates the ability of human amniotic fluid stem cells and human dental pulp stem cell to differentiate into insulin-producing cells, offering a non-pancreatic, low-invasive source of cells for islet regeneration. Copyright © 2013 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.

  7. Propagation of Human Embryonic Stem Cells on Human Amniotic Fluid Cells as Feeder Cells in Xeno-Free Culture Conditions.

    Science.gov (United States)

    Jung, Juwon; Baek, Jin Ah; Seol, Hye Won; Choi, Young Min

    2016-03-01

    Human embryonic stem cells (hESCs) have been routinely cultured on mouse embryonic fibroblast feederlayers with a medium containing animal materials. For clinical application of hESCs, animal-derived products from the animal feeder cells, animal substrates such as gelatin or Matrigel and animal serum are strictly to be eliminated in the culture system. In this study, we performed that SNUhES32 and H1 were cultured on human amniotic fluid cells (hAFCs) with KOSR XenoFree and a humanized substrate. All of hESCs were relatively well propagated on hAFCs feeders with xeno-free conditions and they expressed pluripotent stem cell markers, alkaline phosphatase, SSEA-4, TRA1-60, TRA1-81, Oct-4, and Nanog like hESCs cultured on STO or human foreskin fibroblast feeders. In addition, we observed the expression of nonhuman N-glycolylneuraminic acid (Neu5GC) molecules by flow cytometry, which was xenotransplantation components of contamination in hESCs cultured on animal feeder conditions, was not detected in this xeno-free condition. In conclusion, SNUhES32 and H1 could be maintained on hAFCs for humanized culture conditions, therefore, we suggested that new xenofree conditions for clinical grade hESCs culture will be useful data in future clinical studies.

  8. Cryopreserved human amniotic membrane injection for plantar fasciitis: a randomized, controlled, double-blind pilot study.

    Science.gov (United States)

    Hanselman, Andrew E; Tidwell, John E; Santrock, Robert D

    2015-02-01

    Treatment options for plantar fasciitis have resulted in varied patient outcomes. The aim of this study was to compare a novel treatment, cryopreserved human amniotic membrane (c-hAM), to a traditional treatment, corticosteroid. Our hypothesis was that c-hAM would be safe and comparable to corticosteroids for plantar fasciitis in regard to patient outcomes. A randomized, controlled, double-blind, single-center pilot study was completed. Patients were randomized into one of 2 treatment groups: c-hAM or corticosteroid. Patients received an injection at their initial baseline visit with an option for a second injection at their first 6-week follow-up. Total follow-up was obtained for 12 weeks after the most recent injection. The primary outcome measurement was the Foot Health Status Questionnaire (FHSQ). The secondary outcome measurements were the Visual Analog Scale (VAS) and verbally reported percentage improvement. Data were analyzed between groups for the 2 different cohorts (1 injection versus 2 injections). Twenty-three patients had complete follow-up. Fourteen were randomized to receive corticosteroid and 9 were randomized to receive c-hAM. Three patients in each group received second injections. With the numbers available, the majority of outcome measurements showed no statistical difference between groups. The corticosteroid did, however, have greater FHSQ shoe fit improvement (P = .0244) at 6 weeks, FHSQ general health improvement (P = .0132) at 6 weeks, and verbally reported improvement (P = .041) at 12 weeks in the one-injection cohort. Cryopreserved hAM had greater FHSQ foot pain improvement (P = .0113) at 18 weeks in the 2-injection cohort. Cryopreserved hAM injection may be safe and comparable to corticosteroid injection for treatment of plantar fasciitis. This is a pilot study and requires further investigation. Level I, prospective randomized trial. © The Author(s) 2014.

  9. Fetal antigen 2: an amniotic protein identified as the aminopropeptide of the alpha 1 chain of human procollagen type I

    DEFF Research Database (Denmark)

    Teisner, B; Rasmussen, H B; Højrup, P

    1992-01-01

    with the aminopropeptide of the alpha 1 chain of human procollagen type I as determined by nucleotide sequences. After oxidative procedures normally employed for radio-iodination (iodogen and chloramine-T), FA2 lost its immunoreactivity. An antigen which cross-reacted with polyclonal rabbit anti-human FA2 was demonstrated...... in fetal calf serum. Gel filtration with analysis of fractions by inhibition ELISA showed that the bovine homologue was present in the same molecular forms as those in human amniotic fluid, and immunohistochemical analysis with anti-human FA2 showed that its distribution in bovine skin was identical...... to that of FA2 in human skin. FA2 is a circulating form of the aminopropeptide of the alpha 1 chain of procollagen type I, and this is the first description of its isolation and structural characterization in humans. Udgivelsesdato: 1992-Dec...

  10. Effect of amalgam fillings on the mercury concentration in human amniotic fluid.

    Science.gov (United States)

    Luglie, Pier Franca; Campus, Guglielmo; Chessa, Giannina; Spano, Giovanni; Capobianco, Giampiero; Fadda, Giovanni Maria; Dessole, Salvatore

    2005-02-01

    Methyl mercury (MeHg) and metallic Hg are well known as neurotoxic agents. Dental amalgam contributes significantly to elemental Hg vapour exposure in the general population. There is little information about Hg concentration in human amniotic fluid (AF) of pregnant women and its potential toxic effect on the fetuses. Primary to assess the relationship between the presence of detectable mercury (Hg) concentration in human AF, number and surface areas of amalgam fillings of pregnant women; secondary to analyse their obstetric history and perinatal complications. Seventy-two pregnant women took part in this prospective study. One dentist recorded the dental status, presence, number and surface areas of amalgam fillings. Total Hg concentration in AF was determined in digested samples using automatic cold vapour atomic absorption equipment. The detection limit of Hg in AF, determined from blank readings, was 0.08 ng/ml. To estimate the dependence of the explanatory variables (such as number and surface areas of amalgam fillings, fish consumption, presence of liver or neurological diseases and smoking habits) on mercury concentration several linear regression models were built up. Stepwise logistic regression procedures were running on total sample and on patients with at least one amalgam filling (Positive Filling group = PF). Principal component analysis (PCA) provided two factors, which explained for more the 60% of the variance among the variables. The overall mean Hg concentration in AF among all patients was 0.37+/-0.49 ng/ml. Nineteen (26.4%) women had a Hg concentration or = 0.08 ng/ml (Hg positive group), the mean value of Hg was 0.49+/-0.52 ng/ml. The average number of amalgam fillings was 2.26 +/- 3.19 in the Hg negative group and 5.32+/-3.03 in the Hg positive group (ANOVA one-way p=0.04). A dependence of mercury concentration on number of amalgam fillings (p=0.03), surface area of the amalgam fillings (p=0.04) and fish consumption (p=0.04) was observed but

  11. 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

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

    Science.gov (United States)

    Orczyk-Pawilowicz, Magdalena; Jawien, Ewa; Deja, Stanislaw; Hirnle, Lidia; Zabek, Adam; Mlynarz, Piotr

    2016-01-01

    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.

  13. Gender-Typed Play and Amniotic Testosterone

    Science.gov (United States)

    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…

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

    Science.gov (United States)

    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.

  15. Amniotic fluid embolism

    National Research Council Canada - National Science Library

    Kaur, Kiranpreet; Bhardwaj, Mamta; Kumar, Prashant; Singhal, Suresh; Singh, Tarandeep; Hooda, Sarla

    2016-01-01

    Amniotic fluid embolism (AFE) is one of the catastrophic complications of pregnancy in which amniotic fluid, fetal cells, hair, or other debris enters into the maternal pulmonary circulation, causing cardiovascular collapse...

  16. Human amniotic epithelial cell feeder layers maintain iPS cell pluripotency by inhibiting endogenous DNA methyltransferase 1.

    Science.gov (United States)

    Chen, Qing; Qiu, Chaolin; Huang, Yongyi; Jiang, Lizhen; Huang, Qin; Guo, Lihe; Liu, Te

    2013-11-01

    Maintaining induced pluripotent stem (iPS) cells in an undifferentiated, self-renewing state during long-term cultivation is, at present, a major challenge. We previously showed that human amniotic epithelial cells (HuAECs) were able to provide a good source of feeder cells for mouse and human embryonic or spermatogonial stem cells; however, the epigenetic mechanisms have not been elucidated. In the present study, mouse embryonic fibroblasts (MEFs) and HuAECs were compared as feeder layers for the long-term culture of human iPS cells. The HuAEC feeders allowed human iPS cells to maintain a high level of alkaline phosphatase (AP) activity and to express key stem cell markers during long-term subculture whereas the MEF feeders did not,. Moreover, the HuAEC feeders significantly affected the cell cycle regulation of the iPS cells, maintaining them in the resting stage and the early stage of DNA synthesis (G0/G1 stage). Furthermore, the CpG islands of the Nanog and Oct4 promoters were hypomethylated, while the Nanog- and Oct4-specific loci exhibited higher levels of histone H3 acetylation and lower levels of H3K27 trimethylation in iPS cells cultured on HuAECs compared with those cultured on MEFs. The DNA methyltransferase 1 (DNMT1) expression in iPS cells cultured on HuAECs was shown to be lower than in those cultured on MEFs. In addition, DNMT1-silenced human iPS cells were able to maintain pluripotency over long-term culture on MEFs. In combination, these results suggest that endogenous DNMT1 expression in human iPS cells may be regulated by HuAEC feeder cells and that Nanog and Oct4 are crucial components required for the maintenance of iPS cells in an undifferentiated, proliferative state, capable of self-renewal.

  17. Cloned, CD117 selected human amniotic fluid stem cells are capable of modulating the immune response.

    Directory of Open Access Journals (Sweden)

    Emily C Moorefield

    Full Text Available Amniotic fluid stem (AFS cells are broadly multipotent, can be expanded extensively in culture, are not tumorigenic and can be readily cryopreserved for cell banking. Mesenchymal stem cells (MSC show immunomodulatory activity and secrete a wide spectrum of cytokines and chemokines that suppress inflammatory responses, block mixed lymphocyte reactions (MLR and other immune reactions, and have proven therapeutic against conditions such as graft-versus-host disease. AFS cells resemble MSCs in many respects including surface marker expression and differentiation potential. We therefore hypothesized that AFS cells may exhibit similar immunomodulatory capabilities. We present data to demonstrate that direct contact with AFS cells inhibits lymphocyte activation. In addition, we show that cell-free supernatants derived from AFS cells primed with total blood monocytes or IL-1β, a cytokine released by monocytes and essential in mediation of the inflammatory response, also inhibited lymphocyte activation. Further investigation of AFS cell-free supernatants by protein array revealed secretion of multiple factors in common with MSCs that are known to be involved in immune regulation including growth related oncogene (GRO and monocyte chemotactic protein (MCP family members as well as interleukin-6 (IL-6. AFS cells activated by PBMCs released several additional cytokines as compared to BM-MSCs, including macrophage inflammatory protein-3α (MIP-3α, MIP-1α and Activin. AFS cells also released higher levels of MCP-1 and lower levels of MCP-2 compared to BM-MSCs in response to IL-1β activation. This suggests that there may be some AFS-specific mechanisms of inhibition of lymphocyte activation. Our results indicate that AFS cells are able to suppress inflammatory responses in vitro and that soluble factors are an essential component in the communication between lymphocytes and AFS cells. Their extensive self-renewal capacity, possibility for banking and

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

    Directory of Open Access Journals (Sweden)

    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.

  19. 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.

  20. Comparison of human amniotic membrane and hyaluronate/carboxymethylcellulose membrane for prevention of adhesion formation in rats.

    Science.gov (United States)

    Kelekci, Sefa; Uygur, Dilek; Yilmaz, Bulent; Sut, Necdet; Yesildaglar, Narter

    2007-10-01

    To investigate the effectiveness of human amniotic membrane (HAM) in the prevention of postoperative adhesion formation and to compare it with the efficacy of hyaluronate/carboxymethylcellulose (HA/CMC) membrane in a rat model. Following pilot studies and computer-generated randomization, 23 female Wistar albino rats were operated on in the full study. One of the uterine horns with standard lesions was treated with either HAM (n = 13) or HA/CMC (n = 10) and the other uterine horn served as the control. Second look laparotomies were performed 2 weeks after the operations. Main outcome measures were extent, severity, degree, total adhesion scores and histopathologic characteristics of adhesions. Uterine horns treated with HAM had significantly lower total adhesion scores than the controls (5.15 +/- 2.67 vs. 7.92 +/- 1.50, P CMC membrane were significantly lower than those of the controls (4.30 +/- 1.95 vs. 7.50 +/- 1.84, P CMC groups regarding any adhesion scores. HAM and HA/CMC membrane are both effective for prevention of adhesion formation in a rat uterine horn model; however, one does not seem to be more effective than the other.

  1. 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.

  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. Transplantation of Human Amniotic Mesenchymal Stem Cells Promotes Functional Recovery in a Rat Model of Traumatic Spinal Cord Injury.

    Science.gov (United States)

    Zhou, Hong-Long; Zhang, Xue-Jun; Zhang, Mao-Ying; Yan, Zhong-Jie; Xu, Zhi-Min; Xu, Ru-Xiang

    2016-10-01

    Human amniotic membrane mesenchymal stem cells (hAMSCs) are considered ideal candidate stem cells for cell-based therapy. In this study, we assessed whether hAMSCs transplantation promotes neurological functional recovery in rats after traumatic spinal cord injury (SCI). In addition, the potential mechanisms underlying the possible benefits of this therapy were investigated. Female Sprague-Dawley rats were subjected to SCI using a weight drop device and then hAMSCs, or phosphate-buffered saline (PBS) were immediately injected into the contused dorsal spinal cord at 2 mm rostral and 2 mm caudal to the injury site. Our results indicated that transplanted hAMSCs migrated in the host spinal cord without differentiating into neuronal or glial cells. Compared with the control group, hAMSCs transplantation significantly decreased the numbers of ED1+ macrophages/microglia and caspase-3+ cells. In addition, hAMSCs transplantation significantly increased the levels of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) in the injured spinal cord, and promoted both angiogenesis and axonal regeneration. These effects were associated with significantly improved neurobehavioral recovery in the hAMSCs transplantation group. These results show that transplantation of hAMSCs provides neuroprotective effects in rats after SCI, and could be candidate stem cells for the treatment of SCI.

  4. No midterm advantages in the middle term using small intestinal submucosa and human amniotic membrane in Achilles tendon transverse tenotomy.

    Science.gov (United States)

    Liu, Yushu; Peng, Yinbo; Fang, Yong; Yao, Min; Redmond, Robert W; Ni, Tao

    2016-11-24

    The study was aimed to compare the effects of small intestinal submucosa (SIS) and human amniotic membrane (HAM) on Achilles tendon healing. A total of 48 New Zealand white rabbits were divided into two groups. A full-thickness transverse tenotomy was made at the right leg of the rabbits. Then, the laceration site was wrapped with HAM (P/A group) or SIS (P/S group). The ultimate stress (US) and Young's modulus (E) of the tendons were detected for biomechanical analysis. Histological evaluation was performed using hematoxylin and eosin, immunohistochemical, and immunofluorescent stain. Expression of collagen I was detected by western blot analysis, and levels of inflammatory cytokines IL-1β, IL-6, and TNF-α were measured. Finally, adhesion formation was evaluated. There were no significant differences in filamentous adhesion, cross-sectional areas of the laceration sites, levels of inflammatory response, and collagen type I expression between the P/A and P/S groups (p > 0.05). Compared with the P/A group, the US and E values were significantly higher in the P/S group at day 7 (p Achilles tendon injury in the early stage of healing.

  5. Study of p53 expression and post-transcriptional modifications after GSM-900 radiofrequency exposure of human amniotic cells.

    Science.gov (United States)

    Bourthoumieu, Sylvie; Magnaudeix, Amandine; Terro, Faraj; Leveque, Philippe; Collin, Alice; Yardin, Catherine

    2013-01-01

    The potential effects of radiofrequency (RF) exposure on the genetic material of cells are very important to determine since genome instability of somatic cells may be linked to cancer development. In response to genetic damage, the p53 protein is activated and can induce cell cycle arrest allowing more time for DNA repair or elimination of damaged cells through apoptosis. The objective of this study was to investigate whether the exposure to RF electromagnetic fields, similar to those emitted by mobile phones of the second generation standard, Global System for Mobile Communications (GSM), may induce expression of the p53 protein and its activation by post-translational modifications in cultured human cells. The potential induction of p53 expression and activation by GSM-900 was investigated after in vitro exposure of human amniotic cells for 24 h to average specific absorption rates (SARs) of 0.25, 1, 2, and 4 W/kg in the temperature range of 36.3-39.7 °C. The exposures were carried out using a wire-patch cell (WPC) under strictly controlled conditions of temperature. Expression and activation of p53 by phosphorylation at serine 15 and 37 were studied using Western blot assay immediately after three independent exposures of cell cultures provided from three different donors. Bleomycin-exposed cells were used as a positive control. According to our results, no significant changes in the expression and activation of the p53 protein by phosphorylation at serine 15 and 37 were found following exposure to GSM-900 for 24 h at average SARs up to 4 W/kg in human embryonic cells. Copyright © 2012 Wiley Periodicals, Inc.

  6. Application of Human Amniotic Membrane in Canine Penile Tunica Albuginea Defect: First Step toward an Innovating New Method for Treatment of Peyronie?s Disease

    Directory of Open Access Journals (Sweden)

    M. Salehipour

    2014-06-01

    Full Text Available Purposes To evaluate the efficacy of human amniotic membrane (AM grafting in the canine penile tunica albuginea defect; we developed an animal model as the first step toward an innovating new method for the treatment of Peyronie’s disease, penile cancers, and congenital deformities of the penis. Material and Methods From August to September 2011, ten healthy male dogs were selected. A rhomboid incision about 3x2cm over the tunica albuginea and its overlying squamous epithelium was made and then excised. The amniotic membrane was folded twice on itself and grafted on the defect. After 8 weeks, artificial erection was made for 5 dogs and for the other 5 dogs after 12 weeks. After artificial erection, partial penectomy was done and histopathological evaluation was performed on the grafts. Results Artificial erection performed successfully in all of the dogs. No infection or any other complication was seen. Histopathological examination showed complete re-epithelialization with squamous epithelium and collagen fiber deposition. Also, no dysplasia was seen. Conclusions The amniotic membrane can be used as a suitable substitution for tunica albuginea. It is safe, inexpensive, biodegradable, and available and may be used for the treatment of Peyronie’s disease, penile cancers, congenital penile deformities, and penile reconstructive surgery.

  7. Human amniotic epithelial cells inhibit growth of epithelial ovarian cancer cells via TGF‑β1-mediated cell cycle arrest.

    Science.gov (United States)

    Bu, Shixia; Zhang, Qiuwan; Wang, Qian; Lai, Dongmei

    2017-11-01

    It is reported that human amniotic epithelial cells (hAECs) endow intrinsic antitumor effects on certain kinds of cancer. This research was designed to evaluate whether hAECs endowed potential anticancer properties on epithelial ovarian cancer (EOC) cells in vivo and in vitro, which has not been reported before. In this study, we established a xenografted BALB/c nude mouse model by subcutaneously co-injecting ovarian cancer cell line, SK-OV-3, and hAECs for 28 days. In ex vivo experiments, CCK‑8 cell viability assay, real-time PCR, cell counting assay, cell cycle analysis and immunohistochemistry (IHC) assay were used to detect the effects of hAEC‑secreted factors on the proliferation and cell cycle progression of EOC cells. A cytokine array was conducted to detect anticancer-related cytokines released from hAECs. Human recombinant TGF‑β1 and TGF‑β1 antibody were used to treat EOC cells and analyzed whether TGF‑β1 contributed to the cell cycle arrest. Results from in vivo and ex vivo experiments showed that hAEC-secreted factors and rhTGF‑β1 decreased proliferation of EOC cells and induced G0/G1 cell cycle arrest in cancer cells, which could be partially reversed by excess TGF‑β1 antibody. These data indicate that hAECs endow potential anticancer properties on epithelial ovarian cancer in vivo and in vitro which is partially mediated by hAEC‑secreted TGF‑β1-induced cell cycle arrest. This study suggests a potential application of hAEC‑based therapy against epithelial ovarian cancer.

  8. Liver-Directed Human Amniotic Epithelial Cell Transplantation Improves Systemic Disease Phenotype in Hurler Syndrome Mouse Model.

    Science.gov (United States)

    Rodriguez, Natalie S; Yanuaria, Lisa; Parducho, Kevin Murphy R; Garcia, Irving M; Varghese, Bino A; Grubbs, Brendan H; Miki, Toshio

    2017-07-01

    Mucopolysaccharidosis type 1 (MPS1) is an inherited lysosomal storage disorder caused by a deficiency in the glycosaminoglycan (GAG)-degrading enzyme α-l-iduronidase (IDUA). In affected patients, the systemic accumulation of GAGs results in skeletal dysplasia, neurological degeneration, multiple organ dysfunction, and early death. Current therapies, including enzyme replacement and bone marrow transplant, improve life expectancy but the benefits to skeletal and neurological phenotypes are limited. In this study, we tested the therapeutic efficacy of liver-directed transplantation of a placental stem cell, which possesses multilineage differentiation potential, low immunogenicity, and high lysosomal enzyme activity. Unfractionated human amniotic epithelial cells (hAECs) were transplanted directly into the liver of immunodeficient Idua knockout mouse neonates. The hAECs engraftment was immunohistochemically confirmed with anti-human mitochondria staining. Enzyme activity assays indicated that hAECs transplantation restored IDUA function in the liver and significantly decreased urinary GAG excretion. Histochemical and micro-computed tomography analyses revealed reduced GAG deposition in the phalanges joints and composition/morphology improvement of cranial and facial bones. Neurological assessment in the hAEC treated mice showed significant improvement of sensorimotor coordination in the hAEC treated mice compared to untreated mice. Results confirm that partial liver cell replacement with placental stem cells can provide long-term (>20 weeks) and systemic restoration of enzyme function, and lead to significant phenotypic improvement in the MPS1 mouse model. This preclinical data indicate that liver-directed placental stem cell transplantation may improve skeletal and neurological phenotypes of MPS1 patients. Stem Cells Translational Medicine 2017;6:1583-1594. © 2017 The Authors Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of Alpha

  9. 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.

  10. Human amniotic fluid stem cells do not differentiate into dopamine neurons in vitro or after transplantation in vivo.

    Science.gov (United States)

    Donaldson, Angela E; Cai, Jingli; Yang, Ming; Iacovitti, Lorraine

    2009-09-01

    Although embryonic stem (ES) cells can generate dopamine (DA) neurons that are potentially useful as a cell replacement therapy in Parkinson's disease (PD), associated ethical and practical concerns remain major stumbling blocks to their eventual use in humans. In this study, we examined human amniotic fluid stem (hAFS) cells derived from routine amniocenteses for their potential to give rise to DA neurons in vitro and following transplantation into the 6-hydroxydopamine-lesioned rat brain. We show that undifferentiated hAFS cells constitutively expressed mRNAs and proteins typical of stem cells but also cell derivatives of all three germ layers, including neural progenitors/neurons (nestin, beta-tubulin III, neurofilament). Additionally, these cells expressed mRNAs of an immature DA phenotype (Lmx1a, Pitx-3, Nurr1, Aldh1a1) but not the corresponding proteins. Importantly, treatment with DA differentiation factors using a variety of protocols did not further promote the development of fully differentiated DA neurons from hAFS cells. Thus, Lmx1a, Aldh1a1, AADC, TH, and DAT proteins were not detected in hAFS cells in culture or after transplantation into the PD rat brain. Moreover, by 3 weeks after implantation, there were no surviving AFS cells in the graft, likely as a result of an acute immunorejection response, as evidenced by the abundant presence of CD11+ macrophage/microglia and reactive GFAP+ astrocytes in the host brain. Taken together, these results suggest that further studies will be needed to improve differentiation procedures in culture and to prolong cell survival in vivo if hAFS cells are to be useful as replacement cells in PD.

  11. 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.

  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. Amniotic band syndrome.

    Science.gov (United States)

    Shetty, Prathvi; Menezes, Leo Theobald; Tauro, Leo Francis; Diddigi, Kumar Arun

    2013-10-01

    Amniotic band syndrome is an uncommon congenital disorder without any genetic or hereditary disposition. It involves fetal entrapment in strands of amniotic tissue and causes an array of deletions and deformations. Primary treatment is plastic and reconstructive surgery after birth with in utero fetal surgery also coming in vogue.

  14. Amniotic Band Syndrome

    OpenAIRE

    Shetty, Prathvi; Menezes, Leo Theobald; Tauro, Leo Francis; Diddigi, Kumar Arun

    2012-01-01

    Amniotic band syndrome is an uncommon congenital disorder without any genetic or hereditary disposition. It involves fetal entrapment in strands of amniotic tissue and causes an array of deletions and deformations. Primary treatment is plastic and reconstructive surgery after birth with in utero fetal surgery also coming in vogue.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2007-11-15

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

  16. Synergic activation of toll-like receptor (TLR 2/6 and 9 in response to Ureaplasma parvum & urealyticum in human amniotic epithelial cells.

    Directory of Open Access Journals (Sweden)

    Martha Triantafilou

    Full Text Available Ureaplasma species are the most frequently isolated microorganisms inside the amniotic cavity and have been associated with spontaneous abortion, chorioamnionitis, premature rupture of the membranes (PROM, preterm labour (PL pneumonia in neonates and bronchopulmonary dysplasia in neonates. The mechanisms by which Ureaplasmas cause such diseases remain unclear, but it is believed that inappropriate induction of inflammatory responses is involved, triggered by the innate immune system. As part of its mechanism of activation, the innate immune system employs germ-lined encoded receptors, called pattern recognition receptors (PRRs in order to "sense" pathogens. One such family of PRRs are the Toll like receptor family (TLR. In the current study we aimed to elucidate the role of TLRs in Ureaplasma-induced inflammation in human amniotic epithelial cells. Using silencing, as well as human embryonic kidney (HEK transfected cell lines, we demonstrate that TLR2, TLR6 and TLR9 are involved in the inflammatory responses against Ureaplasma parvum and urealyticum serovars. Ureaplasma lipoproteins, such as Multiple Banded antigen (MBA, trigger responses via TLR2/TLR6, whereas the whole bacterium is required for TLR9 activation. No major differences were observed between the different serovars. Cell activation by Ureaplasma parvum and urealyticum seem to require lipid raft function and formation of heterotypic receptor complexes comprising of TLR2 and TLR6 on the cell surface and TLR9 intracellularly.

  17. Human amniotic epithelial cell feeder layers maintain human iPS cell pluripotency via inhibited endogenous microRNA-145 and increased Sox2 expression

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Te, E-mail: liute79@yahoo.com [School of Environmental Science and Engineering, Donghua University, Shanghai 201620 (China); Shanghai Geriatric Institute of Chinese Medicine, Shanghai 200031 (China); Cheng, Weiwei [International Peace Maternity and Child Health Hospital, Shanghai Jiaotong University, Shanghai 200030 (China); Huang, Yongyi [Laboratoire PROTEE, Batiment R, Universite du Sud Toulon-Var, 83957 LA GARDE Cedex (France); Huang, Qin; Jiang, Lizhen [Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China); Guo, Lihe, E-mail: liute79@yahoo.com [Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031 (China)

    2012-02-15

    Currently, human induced pluripotent stem (iPS) cells were generated from patient or disease-specific sources and share the same key properties as embryonic stem cells. This makes them attractive for personalized medicine, drug screens or cellular therapy. Long-term cultivation and maintenance of normal iPS cells in an undifferentiated self-renewing state are a major challenge. Our previous studies have shown that human amniotic epithelial cells (HuAECs) could provide a good source of feeder cells for mouse and human embryonic stem cells, or spermatogonial stem cells, but the mechanism for this is unknown. Here, we examined the effect of endogenous microRNA-145 regulation on Sox2 expression in human iPS cells by HuAECs feeder cells regulation, and in turn on human iPS cells pluripotency. We found that human IPS cells transfected with a microRNA-145 mutant expressed Sox2 at high levels, allowing iPS to maintain a high level of AP activity in long-term culture and form teratomas in SCID mice. Expression of stem cell markers was increased in iPS transfected with the microRNA-145 mutant, compared with iPS was transfected with microRNA-145. Besides, the expression of Drosha proteins of the microRNA-processor complex, required for the generation of precursor pre-miRNA, was significantly increased in human iPS cells cultured on MEF but not on HuAECs. Taken together, these results suggest that endogenous Sox2 expression may be regulated by microRNA-145 in human iPS cells with HuAECs feeder cells, and Sox2 is a crucial component required for maintenance of them in an undifferentiated, proliferative state capable of self-renewal. Highlights: Black-Right-Pointing-Pointer microRNA-145 inhibits Sox2 expression in human iPS cells. Black-Right-Pointing-Pointer microRNA-145 suppresses the self-renewal and pluripotency of human iPS cells. Black-Right-Pointing-Pointer HuAECs regulate expression of microRNA-145 and Sox2 in human iPS cells. Black-Right-Pointing-Pointer HuAECs feeder

  18. Usos de la membrana amniótica humana en oftalmología Some uses of human amniotic membrane in Ophthalmology

    Directory of Open Access Journals (Sweden)

    Jaime Alemañy González

    2006-06-01

    Full Text Available Se realizó una revisión bibliográfica sobre el uso de la membrana amniótica en oftalmología con los propósitos de conocer sus mecanismos de acción y aplicaciones, así como los resultados publicados sobre el transplante de membrana amniótica humana en la superficie ocular. Se estudiaron textos y artículos publicados sobre este tipo de tratamiento en las enfermedades y traumas de córnea, conjuntiva, esclera y párpados. Se concluye que es una alternativa terapéutica valida que puede ser de gran utilidad en nuestro medioA literature review was made on the use of amniotic membrane in ophthalmology, with the objective of finding out its mechanisms of action and applications as well as the results of human amniotic membrane transplantation for ocular surface reconstruction. A number of texts and articles published on this type of treatment for cornea, conjunctiva, sclera and eyelids diseases and traumas were studied. It was concluded that this is a valid therapeutical alternative that may be very beneficial in our context

  19. Lim Mineralization Protein 3 Induces the Osteogenic Differentiation of Human Amniotic Fluid Stromal Cells through Kruppel-Like Factor-4 Downregulation and Further Bone-Specific Gene Expression

    Directory of Open Access Journals (Sweden)

    Marta Barba

    2012-01-01

    Full Text Available Multipotent mesenchymal stem cells with extensive self-renewal properties can be easily isolated and rapidly expanded in culture from small volumes of amniotic fluid. These cells, namely, amniotic fluid-stromal cells (AFSCs, can be regarded as an attractive source for tissue engineering purposes, being phenotypically and genetically stable, plus overcoming all the safety and ethical issues related to the use of embryonic/fetal cells. LMP3 is a novel osteoinductive molecule acting upstream to the main osteogenic pathways. This study is aimed at delineating the basic molecular events underlying LMP3-induced osteogenesis, using AFSCs as a cellular model to focus on the molecular features underlying the multipotency/differentiation switch. For this purpose, AFSCs were isolated and characterized in vitro and transfected with a defective adenoviral vector expressing the human LMP3. LMP3 induced the successful osteogenic differentiation of AFSC by inducing the expression of osteogenic markers and osteospecific transcription factors. Moreover, LMP3 induced an early repression of the kruppel-like factor-4, implicated in MSC stemness maintenance. KLF4 repression was released upon LMP3 silencing, indicating that this event could be reasonably considered among the basic molecular events that govern the proliferation/differentiation switch during LMP3-induced osteogenic differentiation of AFSC.

  20. Amniotic Fluid Embolism

    Science.gov (United States)

    ... type, as well as a complete blood count (CBC) Electrocardiogram (ECG or EKG) to evaluate your heart's ... mayoclinic.org/diseases-conditions/amniotic-fluid-embolism/basics/definition/CON-20035462 . Mayo Clinic Footer Legal Conditions and ...

  1. 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....

  2. Human beta-defensin 1, 2 and 3 production by amniotic epithelial cells with respect to human papillomavirus (HPV) infection, HPV oncogenic potential and the mode of delivery.

    Science.gov (United States)

    Szukiewicz, Dariusz; Alkhalayla, Habib; Pyzlak, Michal; Watroba, Mateusz; Szewczyk, Grzegorz; Wejman, Jaroslaw

    2016-08-01

    Human beta-defensins (HBD) produced by human amniotic epithelial cells (HAEC) co-create an innate antiviral immune response in the materno-placento-fetal unit. Oncogenic potential of HPV may reflect its ability to avoid immune recognition. In this study we assessed the risk of HAEC infection with human papillomavirus (HPV) in relation to the type of labor and the impact of the oncogenic potential of HPV on HBD production in HAEC. A comparative analysis [HPV(+) vs. HPV(-)HAEC] of the production of HBD were performed. HAEC were isolated from placentas of 116 HPV(+) and 36 HPV(-) parturients (groups I and II, respectively) using trypsin-based method. The cases of premature rupture of membranes (PROM), natural labors (NL) and cesarean sections (CS) were analysed in respective subgroups. High-risk (HR-HPV) and low-risk (LR-HPV) genotypes of HPV in cervical smears and HAEC were identified using the Roche Linear Array(®) HPV Genotyping Test. HBD-1,-2,-3 concentrations in the HAEC culture supernatant were assessed using ELISA. The highest percentage (42.1%) of HPV transmission to HAEC occurred in PROM, an intermediate value was observed after NL (38.5%), and the lowest (25.6%) after CS. The mean concentrations of HBD-2 and HBD-3 in group I were up to 3.1- and 2.8-fold higher (p potential of HBD-2 for HR-HPV infection should be assessed in future studies. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Conditioned medium from human amniotic epithelial cells may induce the differentiation of human umbilical cord blood mesenchymal stem cells into dopaminergic neuron-like cells.

    Science.gov (United States)

    Yang, Shu; Sun, Hai-Mei; Yan, Ji-Hong; Xue, Hong; Wu, Bo; Dong, Fang; Li, Wen-Shuai; Ji, Feng-Qing; Zhou, De-Shan

    2013-07-01

    Dopaminergic (DA) neuron therapy has been established as a new clinical tool for treating Parkinson's disease (PD). Prior to cell transplantation, there are two primary issues that must be resolved: one is the appropriate seed cell origin, and the other is the efficient inducing technique. In the present study, human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) were used as the available seed cells, and conditioned medium from human amniotic epithelial cells (ACM) was used as the inducing reagent. Results showed that the proportion of DA neuron-like cells from hUCB-MSCs was significantly increased after cultured in ACM, suggested by the upregulation of DAT, TH, Nurr1, and Pitx3. To identify the process by which ACM induces DA neuron differentiation, we pretreated hUCB-MSCs with k252a, the Trk receptor inhibitor of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF), and found that the proportion of DA neuron-like cells was significantly decreased compared with ACM-treated hUCB-MSCs, suggesting that NGF and BDNF in ACM were involved in the differentiation process. However, we could not rule out the involvement of other unidentified factors in the ACM, because ACM + k252a treatment does not fully block DA neuron-like cell differentiation compared with control. The transplantation of ACM-induced hUCB-MSCs could ameliorate behavioral deficits in PD rats, which may be associated with the survival of engrafted DA neuron-like cells. In conclusion, we propose that hUCB-MSCs are a good source of DA neuron-like cells and that ACM is a potential inducer to obtain DA neuron-like cells from hUCB-MSCs in vitro for an ethical and legal cell therapy for PD. Copyright © 2013 Wiley Periodicals, Inc.

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

    Science.gov (United States)

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

    2017-01-20

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

  5. Amniotic fluid embolism

    Directory of Open Access Journals (Sweden)

    Kiranpreet Kaur

    2016-01-01

    Full Text Available Amniotic fluid embolism (AFE is one of the catastrophic complications of pregnancy in which amniotic fluid, fetal cells, hair, or other debris enters into the maternal pulmonary circulation, causing cardiovascular collapse. Etiology largely remains unknown, but may occur in healthy women during labour, during cesarean section, after abnormal vaginal delivery, or during the second trimester of pregnancy. It may also occur up to 48 hours post-delivery. It can also occur during abortion, after abdominal trauma, and during amnio-infusion. The pathophysiology of AFE is not completely understood. Possible historical cause is that any breach of the barrier between maternal blood and amniotic fluid forces the entry of amniotic fluid into the systemic circulation and results in a physical obstruction of the pulmonary circulation. The presenting signs and symptoms of AFE involve many organ systems. Clinical signs and symptoms are acute dyspnea, cough, hypotension, cyanosis, fetal bradycardia, encephalopathy, acute pulmonary hypertension, coagulopathy etc. Besides basic investigations lung scan, serum tryptase levels, serum levels of C3 and C4 complements, zinc coproporphyrin, serum sialyl Tn etc are helpful in establishing the diagnosis. Treatment is mainly supportive, but exchange transfusion, extracorporeal membrane oxygenation, and uterine artery embolization have been tried from time to time. The maternal prognosis after amniotic fluid embolism is very poor though infant survival rate is around 70%.

  6. Differentiation of Human Amniotic Mesenchymal Stem Cells into Human Anterior Cruciate Ligament Fibroblast Cells by In Vitro Coculture

    Directory of Open Access Journals (Sweden)

    Yuwan Li

    2017-01-01

    Full Text Available Anterior cruciate ligament injuries are common in humans, though cellular components of the knee have little regenerative or proliferation potential. This study investigated the differentiation of human amnion-derived mesenchymal stem cells (hAMSCs into human anterior cruciate ligament fibroblasts (hACLFs in vitro through induction with bFGF and TGF-β1 with coculture systems. Groups A and B comprised hAMSCs at the 3rd passage cultured with and without bFGF and TGF-β1, respectively; Groups C and D consisted of hAMSCs and hACLFs in monolayer coculture with and without bFGF and TGF-β1, respectively; Groups E and F were composed of hAMSCs and hACLFs in Transwell coculture with and without bFGF and TGF-β1, respectively. Cell morphology and proliferation were recorded. Protein expression and relative mRNA expression were evaluated in each group. Cell proliferation was significantly higher in the induced groups than in the noninduced groups. Protein expression increased over time with the highest expression observed in Group E. mRNA levels were significantly higher in Group E than in other groups. This study is the first to demonstrate the use of the Transwell coculture system for this purpose, and hAMSCs were successfully differentiated into hACLFs. Thus, hAMSCs may be a superior choice for hACLF differentiation via Transwell coculture.

  7. Potential Function of Amniotic Fluid in Fetal Development—Novel Insights by Comparing the Composition of Human Amniotic Fluid with Umbilical Cord and Maternal Serum at Mid and Late Gestation

    Directory of Open Access Journals (Sweden)

    Xing-Long Tong

    2009-07-01

    Conclusion: This study demonstrated that AF is not a result of simple filtration from the blood but an independent fluid. We speculate that proteins or peptides in the amniotic fluid modulate the process of fetus development since they possess potent bioactivity on cellular growth and proliferation. AF provides a pathway to transport these ‘regulators’ to the fetus and thus plays a pivotal role in fetal development.

  8. [An experimental study on the fate of the amniotic membrane after amniotic membrane transplantation for acute alkaline burn of rat cornea].

    Science.gov (United States)

    Chen, You; Yan, Xiao-ming; Wu, Hai-rong; Rong, Bei

    2012-01-01

    Use three different methods of amniotic membrane transplantation for acute alkaline burn of rat cornea, to investigate the difference on operation time, reabsorption time of amniotic membrane, integration patterns of amniotic membrane into the cornea among the three amniotic membrane transplantation methods. SD rats were randomly assigned into five groups. Group A: simply alkaline burn of cornea; Group B: inlay method; Group C: overlay method; Group D: sutureless method; Group E: normal. Prepared the model of corneal alkaline burn of the right eyes of group A-D rats, performed three different methods of amniotic membrane transplantation on Group B-D rats respectively within 24 hours after alkaline burn. The operation time was recorded. Observed the eyes with slit-lamp microscopy after transplantation, recorded the reabsorption time of amniotic membrane. HE staining and immunohistochemical staining against human collagen IV were performed for pathological study. The results were analysed with Image-Pro Plus 6.0 software. The integration patterns of amniotic membrane into the cornea after amniotic membrane reabsorbed were observed. (1) Operation time: Group B was (35.500 ± 2.878) minutes; Group C was (33.375 ± 2.973) minutes; Group D was (9.875 ± 1.246) minutes. There was statistical significance between group B and group D (P cornea after amniotic membrane reabsorption: superficial localization, intraepithelial and intrastromal. We could find one or more integration patterns in same method of amniotic membrane transplantation, there was no statistical significance among group B, C, D (P = 0.86). Amniotic membrane can integrate into the cornea after amniotic membrane reabsorption. Sutureless amniotic membrane transplantation which has short operation time and inconspicuous inflammatory reaction is better than traditional amniotic membrane transplantation, and it will be used extensively in the ophthalmology field in future.

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

    Science.gov (United States)

    Yera, Hélène; Ménégaut, Louise; Brenier-Pinchart, Marie-Pierre; Touafek, Feriel; Bastien, Patrick; Dalle, Frédéric

    2018-01-23

    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®), as compared to three other automated procedures (MagNAPure Compact, Roche®, BioRobot EZ1, Qiagen® and modified NucliSens easyMAG, bioMérieux®) and to 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 1,296 PCRs were performed including 972 Toxoplasma-PCRs. We showed variable efficacy (4.2 to 79.2% positive results) among the DNA extraction procedures in isolating ≤5 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 ≤5 T. gondii cells/mL: when using TaqMan PCR, all the automated systems yielded more than 60% of 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 to the PCR assay used. Copyright © 2018. Published by Elsevier Ltd.

  10. Effect of Amniotic Fluid Stem Cells and Amniotic Fluid Cells on the Wound Healing Process in a White Rat Model

    Directory of Open Access Journals (Sweden)

    Jung Dug Yang

    2013-09-01

    Full Text Available BackgroundAmniotic-fluid-derived stem cells and amniocytes have recently been determined to have wound healing effects, but their mechanism is not yet clearly understood. In this study, the effects of amniotic fluid stem cells and amniocytes on wound healing were investigated through animal experiments.MethodsOn the back of Sprague-Dawley rats, four circular full-thickness skin wounds 2 cm in diameter were created. The wounds were classified into the following four types: a control group using Tegaderm disc wound dressings and experimental groups using collagen discs, amniotic fluid stem cell discs, and amniocyte discs. The wounds were assessed through macroscopic histological examination and immunohistochemistry over a period of time.ResultsThe amniotic fluid stem cell and amniocyte groups showed higher wound healing rates compared with the control group; histologically, the inflammatory cell invasion disappeared more quickly in these groups, and there was more significant angiogenesis. In particular, these groups had significant promotion of epithelial cell reproduction, collagen fiber formation, and angiogenesis during the initial 10 days of the wound healing process. The potency of transforming growth factor-β and fibronectin in the experimental group was much greater than that in the control group in the early stage of the wound healing process. In later stages, however, no significant difference was observed.ConclusionsThe amniotic fluid stem cells and amniocytes were confirmed to have accelerated the inflammatory stage to contribute to an enhanced cure rate and shortened wound healing period. Therefore, they hold promise as wound treatment agents.

  11. Pleiotrophin is involved in the amniotic epithelial cell-induced differentiation of human umbilical cord blood-derived mesenchymal stem cells into dopaminergic neuron-like cells.

    Science.gov (United States)

    Yang, Shu; Xue, Dan-Dan; Wu, Bo; Sun, Hai-Mei; Li, Xiao-Shuang; Dong, Fang; Li, Wen-Shuai; Ji, Feng-Qing; Zhou, De-Shan

    2013-02-28

    We have reported that human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) are capable of differentiating into dopaminergic (DA) neuron-like cells upon being induced by amniotic epithelial cells (AECs). However, what factor(s) is involved in the differentiation process has not been explored out thoroughly. Because pleiotrophin (PTN) is known to exert important trophic effects on DA neurons, in the present study, we investigated whether PTN is released by AECs and whether it is involved in the differentiation of hUCB-MSCs into DA neuron-like cells. The expression and secretion of PTN by AECs were detected by immunofluorescence, RT-PCR and ELISA. The hUCB-MSCs were isolated and treated with AEC-conditioned medium (ACM) or recombinant human PTN. Compared to the controls, a higher proportion of treated cells differentiated into DA neuron-like cells, indicated by the increased expression of TH and DAT and the increased dopamine content. These results indicate that PTN released by AECs acts as a synergetic factor with other neurotrophic factors and is involved in the differentiation of hUCB-MSCs into DA neuron-like cells. We suggest that ACM, which contains PTN and other neurotrophic factors, could potentially be used as an agent to promote the differentiation of DA neuron-like cells from hUCB-MSCs for cell therapy of Parkinson's disease without creating legal or ethical issues. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  12. AMNIOTIC BAND SYNDROME

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    Jovana Đorđević

    2009-04-01

    Full Text Available Amniotic band syndrome (ABS is a set of congenital malformations attributed to amniotic bands that entangle fetal parts during intrauterine life, which results in a broad spectrum of anatomic disturbances - ranging from minor constriction rings and lymphedema of the digits to complex, bizarre multiple congenital anomalies incompatible with life. ABS is not very often, but should be considered in every newborn with congenital anomalies, especially defects of extremities and/or body walls. ABS can be diagnosed prenatally by ultrasound; otherwise, the defects are seen after birth. Child's karyotyping is of great importance, in order to avoid misdiagnosis and incorrect information of recurrence risk. A team of specialists should be included in the treatment and follow-up of children with ABS, according to individual needs of every single patient.The aim of this paper is to point out diagnostic and therapeutic approaches in newborns with ABS trough the report of two cases.

  13. Elemental analysis of human amniotic fluid and placenta by total-reflection X-ray fluorescence and energy-dispersive X-ray fluorescence: child weight and maternal age dependence

    Science.gov (United States)

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

    2001-11-01

    This work is an attempt to evaluate the possible influence of the mother's age in trace element concentrations 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 delivery from healthy mothers and healthy infants and full-term pregnancies. According to the age of the mother, three different groups were considered: 20-25, 25-30 and 30-40 years old. Only two mothers were aged more than 35 years. The weight of the infants ranged from 2.56 to 4.05 kg and three groups were also considered: 2.5-3, 3-3.5 and 3.5-4 kg. The organic matrix of the amniotic fluid samples was removed by treatment with HNO 3 followed by oxygen plasma ashing. Yttrium was used as the internal standard for TXRF analysis. Placenta samples were lyophilized and analyzed by EDXRF without any chemical treatment. Very low levels of Ni and Sr were found in the amniotic fluid samples, and were independent of the age of the mother and weight of the child. Cr, Mn, Se and Pb were at the level of the detection limit. Zn, considered one of the key elements in neonatal health, was not significantly different in the samples analyzed; however, it was weakly related to birth weigh. The concentrations obtained 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 which seemed to be significantly correlated with mother's age and newborn weight were Ca and Fe for both types of sample: Ca levels were increased in heavier children and older mothers; however, Fe increased with increasing maternal age, but decreased for heavier babies. The same conclusions were obtained for placenta and amniotic fluid samples. Cu is closely associated with Fe in its function in the organism

  14. High aminopeptidase N/CD13 levels characterize human amniotic mesenchymal stem cells and drive their increased adipogenic potential in obese women.

    Science.gov (United States)

    Iaffaldano, Laura; Nardelli, Carmela; Raia, Maddalena; Mariotti, Elisabetta; Ferrigno, Maddalena; Quaglia, Filomena; Labruna, Giuseppe; Capobianco, Valentina; Capone, Angela; Maruotti, Giuseppe Maria; Pastore, Lucio; Di Noto, Rosa; Martinelli, Pasquale; Sacchetti, Lucia; Del Vecchio, Luigi

    2013-08-15

    Maternal obesity is associated to increased fetal risk of obesity and other metabolic diseases. Human amniotic mesenchymal stem cells (hA-MSCs) have not been characterized in obese women. The aim of this study was to isolate and compare hA-MSC immunophenotypes from obese (Ob-) and normal weight control (Co-) women, to identify alterations possibly predisposing the fetus to obesity. We enrolled 16 Ob- and 7 Co-women at delivery (mean/SEM prepregnancy body mass index: 40.3/1.8 and 22.4/1.0 kg/m2, respectively), and 32 not pregnant women. hA-MSCs were phenotyped by flow cytometry; several maternal and newborn clinical and biochemical parameters were also measured. The expression of membrane antigen CD13 was higher on Ob-hA-MSCs than on Co-hA-MSCs (P = 0.005). Also, serum levels of CD13 at delivery were higher in Ob- versus Co-pregnant women and correlated with CD13 antigen expression on Ob-hA-MSCs (r2 = 0.84, P women. In conclusion, antigen CD13, by influencing the adipogenic potential of hA-MSCs, could be an in utero risk factor for obesity. Our data strengthen the hypothesis that high levels of serum and MSC CD13 are obesity markers.

  15. Oxidative Stress, DNA, Cell Cycle/Cell Cycle Associated Proteins and Multidrug Resistance Proteins: Targets of Human Amniotic Membrane in Hepatocellular Carcinoma.

    Science.gov (United States)

    Mamede, A C; Guerra, S; Laranjo, M; Santos, K; Carvalho, M J; Carvalheiro, T; Moura, P; Paiva, A; Abrantes, A M; Maia, C J; Botelho, M F

    2016-10-01

    The anticancer effects of human amniotic membrane (hAM) have been studied over the last decade. However, the action mechanisms responsible for these effects are not fully understood until now. Previously results reported by our team proved that hAM is able to induce cytotoxicity and cell death in hepatocellular carcinoma (HCC), a worldwide high incident and mortal cancer. Therefore, this experimental study aimed to investigate the cellular targets of hAM protein extracts (hAMPE) in HCC through in vitro studies. Our results showed that hAMPE is able to modify oxidative stress environment in all HCC cell lines, as well as its cell cycle. hAMPE differently targets deoxyribonucleic acid (DNA), P21, P53, β-catenin and multidrug resistance (MDR) proteins in HCC cell lines. In conclusion, hAMPE has several targets in HCC, being clear that the success of this treatment depends of a personalized therapy based on the biological and genetic characteristics of the tumor.

  16. 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.

  17. Human amniotic membrane, best healing accelerator, and the choice of bone induction for vestibuloplasty technique (an animal study

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    Ahad Khoshzaban

    2010-12-01

    Full Text Available Mohammad H Samandari1, Shahriar Adibi2, Ahad Khoshzaban3, Sara Aghazadeh5, Parviz Dihimi4, Siamak S Torbaghan6, Saeed H Keshel5, Zohreh Shahabi71Department of Oral and Maxillofacial Surgery, Dentistry Faculty, 2Dental Research of Torabinejad Research Centre, 3Iranian Tissue Bank Research and Preparation Centre, Imam Khomeini Hospital Complex, 4Department of Oral and Maxillofacial Pathology, Dentistry Faculty, Isfahan University of Medical Sciences, Isfahan, Iran; 5Stem Cells Preparation Unit, Eye Research Center, Farabi Hospital, 6Department of Pathology, Imam Khomeini Medical Centre, 7BMT Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, IranObjective: To investigate the effects of amniotic membrane (AM in bone induction and wound healing after vestibuloplasty surgery on animal samples while receptacle proteins such as growth factors were considered as accelerators for wound healing and bone induction after these operations.Material and methods: Ten adult dogs (5 females, 5 males; race, Iranian mixed; weight, 44 pounds were included, which underwent surgery for transplantation on mandible and maxillary. AM was used for promoting bone induction and healing.Results: The tissue samples were obtained after 2, 8, and 12 weeks for histology survey. No significant differences were observed between male and female or left and right jaws. AM decreased fibrinoleukocytic exudates and inflammation in the experimental group, had significant effects on bone formation, considerably improves wound healing, and gives rise to bone induction (P < 0.0001.Conclusions: Our study findings indicate that the AM is a suitable cover for different injuries and acellular AM has the potential for rapid improvement and bone induction. The AM contains collagen, laminin, and fibronectin, which provide an appropriate substrate for bone induction. This substrate promoted bone induction and might contribute to induction of the progenitor cells and/or stem

  18. Menaquinone-4 enhances osteogenic potential of human amniotic fluid mesenchymal stem cells cultured in 2D and 3D dynamic culture systems.

    Science.gov (United States)

    Mandatori, Domitilla; Penolazzi, Letizia; Pipino, Caterina; Di Tomo, Pamela; Di Silvestre, Sara; Di Pietro, Natalia; Trevisani, Sara; Angelozzi, Marco; Ucci, Mariangela; Piva, Roberta; Pandolfi, Assunta

    2017-05-16

    Menaquinones, also known as Vitamin K2 family, regulate calcium homeostasis in a 'bone-vascular cross-talk' and recently received particular attention for their positive effect on bone formation. Given that the correlation between menaquinones and bone metabolism to date is still unclear, the objective of our study was to investigate the possible role of menaquinone-4 (MK-4), an isoform of the menaquinones family, in the modulation of osteogenesis. For this reason, we used a model of human amniotic fluid mesenchymal stem cells (hAFMSCs) cultured both in two-dimensional (2D) and three-dimensional (3D; RCCS™bioreactor) in vitro culture systems. Furthermore, to mimic the 'bone remodelling unit' in vitro, hAFMSCs were co-cultured in the 3D system with human monocyte cells (hMCs) as osteoclast precursors. The results showed that in a conventional 2D culture system, hAFMSCs were responsive to the MK-4, which significantly improved the osteogenic process through γ-glutamyl carboxylase-dependent pathway. The same results were obtained in the 3D dynamic system where MK-4 treatment supported the osteoblast-like formation promoting the extracellular bone matrix deposition and the expression of the osteogenic-related proteins (alkaline phosphatase, osteopontin, collagen type-1 and osteocalcin). Notably, when the hAFMSCs were co-cultured in a 3D dynamic system with the hMCs, the presence of MK-4 supported the cellular aggregate formation as well as the osteogenic function of hAFMSCs, but negatively affected the osteoclastogenic process. Taken together, our results demonstrate that MK-4 supported the aggregate formation of hAFMSCs and increased the osteogenic functions. Specifically, our data could help to optimize bone regenerative medicine combining cell-based approaches with MK-4 treatment. Copyright © 2017 John Wiley & Sons, Ltd.

  19. Regulation of amniotic fluid volume

    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

  20. Periostin as a Biomarker of the Amniotic Membrane

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    Mariya P. Dobreva

    2012-01-01

    Full Text Available Tracing the precise developmental origin of amnion and amnion-derived stem cells is still challenging and depends chiefly on analyzing powerful genetic model amniotes like mouse. Profound understanding of the fundamental differences in amnion development in both the disc-shaped primate and human embryo and the cup-shaped mouse embryo is pivotal in particular when sampling amniotic membrane from nonprimate species for isolating candidate amniotic stem cells. The availability of molecular marker genes that are specifically expressed in the amniotic membrane and not in other extraembryonic membranes would be instrumental to validate unequivocally the starting material under investigation. So far such amniotic markers have not been reported. We postulated that bone morphogenetic protein (BMP target genes are putative amniotic membrane markers mainly because deficiency in one of several components of the BMP signaling cascade in mice has been documented to result in defective development of the early amnion. Comparative gene expression analysis of acknowledged target genes for BMP in different extraembryonic tissues, combined with in situ hybridization, identified Periostin (Postn mRNA enrichment in amnion throughout gestation. In addition, we identify and propose a combination of markers as transcriptional signature for the different extraembryonic tissues in mouse.

  1. Decellularized human amniotic membrane: more is needed for an efficient dressing for protection of burns against antibiotic-resistant bacteria isolated from burn patients.

    Science.gov (United States)

    Gholipourmalekabadi, M; Bandehpour, M; Mozafari, M; Hashemi, A; Ghanbarian, H; Sameni, M; Salimi, M; Gholami, M; Samadikuchaksaraei, A

    2015-11-01

    Human amniotic membranes (HAMs) have attracted the attention of burn surgeons for decades due to favorable properties such as their antibacterial activity and promising support of cell proliferation. On the other hand, as a major implication in the health of burn patients, the prevalence of bacteria resistant to multiple antibiotics is increasing due to overuse of antibiotics. The aim of this study was to investigate whether HAMs (both fresh and acellular) are an effective antibacterial agent against antibiotic-resistant bacteria isolated from burn patients. Therefore, a HAM was decellularized and tested for its antibacterial activity. Decellularization of the tissue was confirmed by hematoxylin and eosin (H&E) and 4,6-diamidino-2-phenylindole (DAPI) staining. In addition, the cyto-biocompatibility of the acellular HAM was proven by the cell viability test (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide, MTT) and scanning electron microscopy (SEM). The resistant bacteria were isolated from burns, identified, and tested for their susceptibility to antibiotics using both the antibiogram and polymerase chain reaction (PCR) techniques. Among the isolated bacteria, three blaIMP gene-positive Pseudomonas aeruginosa strains were chosen for their high resistance to the tested antibiotics. The antibacterial activity of the HAM was also tested for Klebsiella pneumoniae (American Type Culture Collection (ATCC) 700603) as a resistant ATCC bacterium; Staphylococcus aureus (mecA positive); and three standard strains of ATCC bacteria including Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27833), and S. aureus (ATCC 25923). Antibacterial assay revealed that only the latter three bacteria were susceptible to the HAM. All the data obtained from this study suggest that an alternative strategy is required to complement HAM grafting in order to fully protect burns from nosocomial infections. Copyright © 2015 Elsevier Ltd and ISBI. All rights reserved.

  2. 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.

  3. Histologic features of transplanted amniotic membrane: implications for corneal wound healing.

    Science.gov (United States)

    Said, Dalia G; Nubile, Mario; Alomar, Thaer; Hopkinson, Andy; Gray, Trevor; Lowe, James; Dua, Harminder S

    2009-07-01

    To evaluate the histologic changes occurring in the transplanted amniotic membrane in human eyes. Observational consecutive case series. Seven consecutive patients who underwent amniotic membrane transplantation (AMT) for bullous keratopathy and subsequently had a penetrating keratoplasty (PK). Corneal buttons obtained at PK were examined by light and electron microscopy and by immunohistology with antibodies against CD34 (keratocytes), alpha smooth muscle actin and vimentin (myofibroblasts and fibroblasts respectively). Time from AMT to PK ranged from 2 to 32 months. Immunophenotypic characteristics of cells populating transplanted amniotic stroma. Amniotic tissue was covered with stratified corneal epithelium with well-defined desmosomes and hemidesmosomes. Transformed corneal stroma-derived cells (CSDCs) could be seen migrating from the anterior stroma, through breaks in the Bowman's zone, into connective tissue of the amniotic membrane. Immunohistology showed that the cells populating amniotic stroma were CD34 negative but positive for vimentin and alpha smooth muscle actin. In 2 samples in which corneal transplants were performed approximately 1 year or more after AMT, some cells in the amniotic stroma showed CD34+ staining. Features of increased metabolic activity and formation of new collagen were seen on electron microscopy. In 2 cases, epithelial cell nests were seen in the amniotic stroma. The amniotic basement membrane facilitates epithelial cell migration and adhesion. The amniotic stroma supports CSDCs and epithelial cells. Repopulation of the amniotic stroma by CSDCs migrating through breaks in Bowman's zone integrates the amnion with corneal tissue and allows for rebuilding of corneal stroma. Over time, some CSDCs may revert to the resting keratocyte immunophenotype.

  4. Amniotic Fluid Stem Cells: Future Perspectives

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    Margit Rosner

    2012-01-01

    Full Text Available The existence of stem cells in human amniotic fluid was reported for the first time almost ten years ago. Since this discovery, the knowledge about these cells has increased dramatically. Today, amniotic fluid stem (AFS cells are widely accepted as a new powerful tool for basic research as well as for the establishment of new stem-cell-based therapy concepts. It is possible to generate monoclonal genomically stable AFS cell lines harboring high proliferative potential without raising ethical issues. Many different groups have demonstrated that AFS cells can be differentiated into all three germ layer lineages, what is of relevance for both, the scientific and therapeutical usage of these cells. Of special importance for the latter is the fact that AFS cells are less tumorigenic than other pluripotent stem cell types. In this paper, we have summarized the current knowledge about this relatively young scientific field. Furthermore, we discuss the relevant future perspectives of this promising area of stem cell research focusing on the next important questions, which need to be answered.

  5. Induction of reprogramming of human amniotic epithelial cells into iPS cells by overexpression of Yap, Oct4, and Sox2 through the activation of the Hippo-Yap pathway.

    Science.gov (United States)

    Zhao, Yanhui; Fei, Xinfeng; Guo, Jianming; Zou, Gang; Pan, Weidong; Zhang, Jingju; Huang, Yongyi; Liu, Te; Cheng, Weiwei

    2017-07-01

    The present study has reported a novel method for producing induced pluripotent stem (iPS) cells. Primary human amniotic epithelial cells (HuAECs) were isolated from the amniotic membranes of pregnant women who received Cesarean sections. These cells were infected with retroviruses carrying octamer-binding transcription factor 4 (Oct4), (sex determining region Y)-box 2 (Sox2) and Yes-associated protein (Yap) (OSY). Following in vitro culture for ~14 days, epithelial-like HuAECs exhibited several iPS clone-like cell colonies (OSY-iPS). These cell clones presented positive alkaline phosphatase features and expressed high levels of embryonic stem cell-like markers (Nanog homeobox, Sox2, Oct4, reduced expression protein 1, and SSES3/4). Additionally, epigenetic analysis results indicated that the methylation of CpG islands on endogenous Oct4 and Sox2 promoters was reduced in OSY-iPS cells. Furthermore, the majority of the histone H3 at lysine 9 sites that interacted with the Oct4 and Sox2 promoters were acetylated, suggesting that the transcription activities of the above two transcription factors significantly increased. In vivo and in vitro induced differentiation experiments demonstrated that OSY-iPS could develop into embryoid bodies in vitro, and express numerous cellular markers in the three germ layers. Furthermore, OSY-iPS could form teratomas in immunodeficient mice. The pathological detection results suggest that these teratomas contain numerous types of cells from the three germ layers. However, the results from the quantitative polymerase chain reaction and western blot analyses suggest that the Hippo-Yap signaling pathway was significantly activated in OSY-iPS cells. In conclusion, a novel method for iPS induction was established in the present study. HuAECs were successfully induced to reprogram iPS cells through the introduction of OSY to activate the Hippo-Yap signaling pathway.

  6. Comparative study between amniotic-fluid mesenchymal stem cells and retinal pigmented epithelium (RPE) stem cells ability to differentiate towards RPE cells.

    Science.gov (United States)

    Mariotti, Cesare; Lazzarini, Raffaella; Nicolai, Michele; Saitta, Andrea; Orsini, Emanuele; Orciani, Monia; Di Primio, Roberto

    2015-10-01

    Dysfunction of the retinal pigmented epithelium (RPE) is one of the first effects of dry age-related macular degeneration (AMD) with consequent blindness. Hence, patients affected by this retinal disorder could benefit from a cell-based transplantation strategy for RPE. Actually, an effective protocol to approach this problem is lacking, though recently, it has been postulated the existence of a subpopulation of RPE stem cells (RPESCs) derived from adult RPE and able to reconstitute a functional RPE. On the other hand, the evidence related to the differentiative potential of human mesenchymal stem cells (MSCs) is continuously increasing. Among others, amniotic fluid-derived MSCs (AF-MSCs) may be a promising candidate, since these cells are characterized by high proliferation and differentiative potential. In this study, AF-MSCs and RPESCs were isolated, characterized to assay their stemness and induced to neuronal/retinal differentiation; specific RPE markers were then analyzed. Our results indicate that RPESCs are more suitable candidates for RPE replacement than AF-MSCs.

  7. Developmental Alterations of α-Fetoprotein Sugar Chain in Amniotic Fluids Analyzed by Lectin Affinity Electrophoresis

    OpenAIRE

    Ohta, Masahiro; Kawahara, Nobuaki; Liu, Miao; Taketa,Kazuhisa; Kudo, Takafumi; Taga, Hiroko

    1998-01-01

    Affinity electrophoresis of human alpha-fetoprotein (AFP) in amniotic fluid from pregnant women between 6 to 42 weeks of gestation and in the serum of a yolk sac tumor was performed with concanavalin A (Con A), lentil lectin (LCA), erythroagglutinating phytohemagglutinin (E-PHA) and Allomyrina dichotoma lectin (allo A). Separated AFP bands were detected by sensitive antibody-affinity blotting. In the first trimester, amniotic fluid AFP showed elevated percentages of Con A-nonreacting AFP (AFP...

  8. Protection of Brain Injury by Amniotic Mesenchymal Stromal Cell-Secreted Metabolites.

    Science.gov (United States)

    Pischiutta, Francesca; Brunelli, Laura; Romele, Pietro; Silini, Antonietta; Sammali, Eliana; Paracchini, Lara; Marchini, Sergio; Talamini, Laura; Bigini, Paolo; Boncoraglio, Giorgio B; Pastorelli, Roberta; De Simoni, Maria-Grazia; Parolini, Ornella; Zanier, Elisa R

    2016-11-01

    To define the features of human amniotic mesenchymal stromal cell secretome and its protective properties in experimental models of acute brain injury. Prospective experimental study. Laboratory research. C57Bl/6 mice. Mice subjected to sham or traumatic brain injury by controlled cortical impact received human amniotic mesenchymal stromal cells or phosphate-buffered saline infused intracerebroventricularly or intravenously 24 hours after injury. Organotypic cortical brain slices exposed to ischemic injury by oxygen-glucose deprivation were treated with human amniotic mesenchymal stromal cells or with their secretome (conditioned medium) in a transwell system. Traumatic brain injured mice receiving human amniotic mesenchymal stromal cells intravenously or intracerebroventricularly showed early and lasting functional and anatomical brain protection. cortical slices injured by oxigen-glucose deprivation and treated with human amniotic mesenchymal stromal cells or conditioned medium showed comparable protective effects (neuronal rescue, promotion of M2 microglia polarization, induction of trophic factors) indicating that the exposure of human amniotic mesenchymal stromal cells to the injured tissue is not necessary for the release of bioactive factors. Using sequential size-exclusion and gel-filtration chromatography, we identified a conditioned medium subfraction, which specifically displays these highly protective properties and we found that this fraction was rich in bioactive molecules with molecular weight smaller than 700 Da. Quantitative RNA analysis and mass spectrometry-based peptidomics showed that the active factors are not proteins or RNAs. The metabolomic profiling of six metabolic classes identified a list of molecules whose abundance was selectively elevated in the active conditioned medium fraction. Human amniotic mesenchymal stromal cell-secreted factors protect the brain after acute injury. Importantly, a fraction rich in metabolites, and

  9. Methods of amniotic membrane fixation in ocular amniotic membrane surgeries

    Directory of Open Access Journals (Sweden)

    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.

  10. 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.

  11. 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

  12. Amniotic membrane transplantation in entropion surgery.

    Science.gov (United States)

    Ti, S E; Tow, S L; Chee, S P

    2001-07-01

    To evaluate the role of amniotic membrane transplantation in the management of cicatricial eyelid entropion. Prospective, noncomparative interventional case series. Eighteen consecutive patients with cicatricial entropion. A gray line lid split procedure with vertical anterior lamella repositioning was performed on 25 eyelids (upper or lower) of 18 patients with moderate to severe cicatricial entropion. Preserved human amniotic membrane (AM) was used to cover the bare tarsus up to the lid margin and secured with running 7-0 Vicryl. Impression cytology of the AM was performed at various stages postoperatively to study the epithelialization process. (1) Reepithelialization of bare tarsus, (2) extent of tarsal shrinkage, (3) recurrence of entropion. All the AM grafts took well. The most common complication was hemorrhage below the graft, which occurred in six cases. Complete success with no lashes touching the globe was achieved in 22 of 25 (88%) lids after a minimum follow-up of 12.0 months. The mean follow-up was 17.8 months. Two cases (qualified success) had recurrent trichiasis treated successfully with electrolysis. One case with severe trachomatous upper lid entropion recurred 14 months after surgery. The AM accelerated the epithelialization of bare tarsus; this was demonstrated by lack of fluorescein staining and reversion to skin color within 2 to 3 weeks. However, AM could not prevent tarsal shrinkage. Impression cytology demonstrated that features of conjunctival epithelium were present for the first postoperative month, but this was gradually replaced by squamous metaplasia, with keratinization appearing as early as 3 weeks postoperatively. The use of AM in a lid split procedure for correction of cicatricial entropion helps the bare tarsus epithelialize rapidly and improves the initial cosmetic result of surgery.

  13. Effect of Amniotic Membrane Combined with Ciprofloxacin in Curing the Primary Stages of Pseudomonal Keratitis

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    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.

  14. Isolation and identification of java race amniotic membrane secretory leukocyte protease inhibitor gene

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    Elly Munadziroh

    2008-09-01

    Full Text Available Background: Secretory leukocyte protease inhibitor (SLPI has been found to facilitate epithelialization, maintain a normal epithelial phenotype, reduce inflammation, secrete growth factors such as IL-4, IL-6, IL-10, EGF, FGF, TGF, HGFand 2-microbulin. SLPI is serine protease inhibitor, which found in secretions such as whole saliva, seminal fluid, cervical mucus, synovial fluid, breast milk, tears, amniotic fluid and amniotic membrane. Impaired healing states are characterized by excessive proteolysis and oftenbacterial infection, leading to the hypothesis that SLPI may have a role in the healing process in oral inflammation and contributes to tissue repair in oral mucosa. The oral wound healing response is impaired in the SLPI sufficient mice since matrix synthesis and collagen deposition delayed. The objective of this research is to isolate and identify the amniotic membrane of Java Race SLPI Gene. Methods: SLPI RNA was isolated from Java Race amniotic membrane and the cDNA was amplified by polymerase chain reaction (PCR. Result: Through sequence analyses, SLPI cDNA was 530 nucleotide in length with a predicted molecular mass about 12 kDa. The nucleotide sequence showed that human SLPI from sample was 98% identical with human SLPI from gene bank. PCR analysis revealed that the mRNA of SLPI was highly expressed in the amniotic membrane from Java Race sample. Conclusion: it is demonstrated that human SLPI are highly conserved in sequence content as compared to the human SLPI from gene.

  15. 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.

  16. Amniotic Fluid Aspiration in Medicolegal Autopsies

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    Sermet Koç

    1999-04-01

    Full Text Available Amniotic fluid aspiration occurs due to intrauterine anoxia. Being one of the causes of respiratory insufficiency and death in newborns, it can be diagnosed solely by histopathological examination. In the present paper, 12 cases with the diagnosis of amniotic fluid aspiration in autopsies between 1996-1997 in the mortuary section of the Council of Forensic Medicine are reevaluated from the medicolegal point of view. In the majority of the cases, delivery took place in locations other than hospitals, and the lack of medical intervention has been an important factor for the infavorable outcome of the autopsies. Findings of amniotic fluid aspiration stained with meconium were detected in four cases. Key words: Amniotic fluid aspiration, intrauterin asphyxia, Neonatal death, Medicolegal autopsy.

  17. 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.

  18. Single and multilayer amniotic membrane transplantation for persistent corneal epithelial defect with and without stromal thinning and perforation

    OpenAIRE

    Prabhasawat, P.; Tesavibul, N.; Komolsuradej, W.

    2001-01-01

    AIMS—To evaluate the efficacy of amniotic membrane transplantation (AMT) in persistent corneal epithelial defect with or without stromal thinning and corneal perforation.
METHODS—28 patients (28 eyes) with persistent corneal epithelial defect unresponsive to medical treatment were given preserved human amniotic membrane transplants. The patients were divided into three groups: group A, persistent corneal epithelial defect 10 eyes; group B, epithelial defect with stromal thinning 13 eyes; and ...

  19. [Advances in preparation and clinical application of amniotic membrane graft].

    Science.gov (United States)

    Zheng, S Q; Chen, T S; Ji, S Z; Luo, P F; Xiao, S C

    2017-08-20

    Amniotic membrane has been used as wound dressing for more than 100 years. With the development of the preservation and preparation techniques, amniotic membrane is widely used in ophthalmology, burns, plastic surgery, dentistry, and neurosurgery. In recent years, as more and more amniotic membrane is used in chronic wounds, it will be a new treatment method for wounds. This paper is a brief review about advances in preparation and clinical application of amniotic membrane graft.

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

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    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.

  1. 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 ...

  2. Effects of ionizing radiation on glycerolated amniotic membranes as a substract for cultured human epithelium; Efeitos da radiacao ionizante em membranas amnioticas gliceroladas empregadas como substrato ao cultivo de epitelio humano

    Energy Technology Data Exchange (ETDEWEB)

    Paggiaro, Andre Oliveira

    2011-07-01

    The amniotic membrane (AM) is a biomaterial with biological properties that are beneficial to tissue repair. It has been used as a temporary coverage to threat burns and chronic wounds. Recently, it has been served as a substrate for keratinocytes culture to construct a living skin equivalent. However, MA is a biological material, and its transplantation could cause infectious disease for receptors. So, it must be preserved and sterilized before clinical use. The aim of this study was to evaluate the radiation effects on glycerol-preserved MA, considering its compatibility to support human keratinocytes culture. Four MA were stored in high concentrations of glycerol (> 85%) and half of them were radio sterilized with a dose of 25 kGy. Then, we established two groups: nonirradiated MA (MA-ni) and irradiated MA (MA-i). Both groups was deepithelialized by a standardized protocol and was investigated morphologically, immunohistochemical and ultrastructural. Subsequently human keratinocytes were cultivated immersed and in air-liquid interface on denuded surface of MA-i and MA-ni. The results were compared at 14 and 21 days of culture by light and electron microscopy. After epithelial denudation, analyses demonstrated the continuity of the basement membrane in MA-ni group, whereas in the irradiated group, there was no indication of the basement membrane’s presence on the surface of MA. The cell cultures showed that in the non-irradiated group, there was growth of a multi-layered and differentiated epithelium, with a stratum corneum’s formation in air-liquid interface. In the irradiated group, the epithelium had only two or three layer, little cell differentiation, with the same results immersed or air-liquid interface system. Glycerol-preserved MA was biocompatible with the growth of a cultivated epithelium, showing its potential as a skin substitute. Irradiation at 25 kGy cause structural damage to the tissue, making changes in basement membrane, that facilitates

  3. Amniotic fl uid embolism in an HIV-positive parturient | Penfold ...

    African Journals Online (AJOL)

    We present a case of a parturient infected with human immunode ciency virus, who developed amniotic fluid embolism during the delivery of her twins by elective Caesarean section. Our management and the available literature are briefly discussed, and consideration is given to a possible association between the two ...

  4. The Paracrine Effect of Transplanted Human Amniotic Epithelial Cells on Ovarian Function Improvement in a Mouse Model of Chemotherapy-Induced Primary Ovarian Insufficiency

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    Xiaofen Yao

    2016-01-01

    Full Text Available Human amnion epithelial cells (hAECs transplantation via tail vein has been reported to rescue ovarian function in mice with chemotherapy-induced primary ovarian insufficiency (POI. To test whether intraperitoneally transplanted hAECs could induce therapeutic effect and to characterize the paracrine effect of transplanted hAECs, we utilized a chemotherapy induced mice model of POI and investigated the ability of hAECs and conditioned medium collected from cultured hAECs (hAECs-CM to restore ovarian function. We found that transplantation of hAECs or hAECs-CM either 24 hours or 7 days after chemotherapy could increase follicle numbers and partly restore fertility. By PCR analysis of recipient mice ovaries, the presence of SRY gene was only detected in mice transplanted with male hAECs 24 hours following chemotherapy. Further, the gene expression level of VEGFR1 and VEGFR2 in the ovaries decreased, although VEGFA increased 2 weeks after chemotherapy. After treatment with hAECs or hAEC-CM, the expression of both VEGFR1 and VEGFR2 increased, consistent with the immunohistochemical analysis. In addition, both hAECs and hAECs-CM treatment enhanced angiogenesis in the ovaries. The results suggested that hAECs-CM, like hAECs, could partly restore ovarian function, and the therapeutic function of intraperitoneally transplanted hAECs was mainly induced by paracrine-mediated ovarian protection and angiogenesis.

  5. Long-term efficacy of glycerine-processed amniotic membrane transplantation in patients with corneal ulcer.

    Science.gov (United States)

    Uhlig, Constantin E; Frings, Charlotte; Rohloff, Nadine; Harmsen-Aasman, Christel; Schmitz, Ralf; Kiesel, Ludwig; Eter, Nicole; Busse, Holger; Alex, Anne F

    2015-09-01

    The aim of this study was to determine the long-term treatment efficacy of glycerine-preserved human amniotic membrane transplantation in patients suffering from corneal ulcers. This was a retrospective, non-controlled, monocentric analysis. Included were patients with corneal ulcers that were non-responsive to ointment or contact lenses and had been treated by amniotic membrane transplantation with either the overlay or sandwich procedure. Analysis parameters were visual acuity before and following treatment, recurrence rate and subjective comfort at the last follow-up. Of the 371 amniotic membrane transplantations that were conducted, 135 surgical treatments in 108 patients (51.9% male, 48.1% female; mean age 63.7 years) met the inclusion criteria. In total, 99 overlay and 36 multilayer amniotic membrane transplantations were performed. The follow-up period was 47.5 ± 66.7 weeks (mean ± SD). The recurrence rate at the last follow-up was 47.8% with overlay membranes and 51.8% with the sandwich technique. There was no significant change in best-corrected visual acuity following treatment with overlays (p = 0.219) or sandwich procedure (p = 0.703). At the last follow-up, 72.1% (overlay) and 78.3% (sandwich) of the patients reported either no pain or increased comfort. The recurrence rates and changes in visual acuity following overlay or sandwich amniotic membrane transplantation in patients suffering from corneal ulcer were lower than reported elsewhere in the literature. More than half of the patients profited from each of the amniotic membrane transplantation techniques with respect to recurrence and postoperative comfort. © 2015 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

  6. Spontaneous Monochorionic Tetra‑amniotic Quadruplet Pregnancy ...

    African Journals Online (AJOL)

    Ron-El R, Mor Z, Weinraub Z, Schreyer P, Bukovsky I, Dolphin Z, et al. Triplet, quadruplet and quintuplet pregnancies: Management and outcome. Acta Obstet Gynaecol Scand 1992;71:347-50. How to cite this article: Nnadi D, Ibrahim A, Nwobodo E. Spontaneous monochorionic tetra-amniotic quadruplet pregnancy at term.

  7. Amniotic amputation | Ayadi | Pan African Medical Journal

    African Journals Online (AJOL)

    Prenatal ultrasonography follow-up showed no abnormalities. Postnatal examination showed signs consistent with the diagnosis of amniotic bands syndrome at the newborn's left limbs. The newborn presented syndactyly, lymphedema, with distal agenesis of the 2nd, 3rd, and 4th fingers; ring constrictions of the great toe, ...

  8. 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.

  9. 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.

  10. Transabdominal collection of amniotic fluid "sludge" and identification of Candida albicans intra-amniotic infection.

    Science.gov (United States)

    Kusanovic, Juan P; Romero, Roberto; Martinovic, Carolina; Silva, Karla; Erez, Offer; Maymon, Eli; Díaz, Francisco; Ferrer, Fernando; Valdés, Rafael; Córdova, Víctor; Vargas, Paula; Nilo, María Elena; Le Cerf, Patricio

    2017-05-03

    A G3P2 patient who conceived while using an intrauterine contraceptive device (IUD) presented at 20 weeks of gestation with mild irregular uterine contractions and vaginal bleeding. Sonographic examination at admission showed the presence of dense amniotic fluid "sludge" and a long sonographic uterine cervix (42 mm). To assess the microbiologic significance of amniotic fluid "sludge", we performed a transabdominal amniocentesis. The procedure was performed under real-time ultrasound, and fluid resembling pus at gross examination was noted. Rapid amniotic fluid analysis showed the presence of a high white blood cell count and structures resembling hyphae. Amniotic fluid cultures were positive for Candida albicans. Treatment was begun with broad-spectrum antibiotics, including Fluconazole, upon the visualization of pus in the "sludge" material because of the presence of hyphae in the Gram stain. Despite treatment, the patient went into spontaneous preterm labor and delivered five days after admission. Placental examination revealed acute fungal histologic chorioamnionitis and funisitis. This represents the first report of transabdominal collection and analysis of amniotic fluid "sludge" and the microbiologic detection of Candida albicans in this material. This report provides evidence that transabdominal retrieval of "sludge" is possible and may be of significant value for patient management and selection of antimicrobial agents.

  11. The origin of fetal sterols in second-trimester amniotic fluid : endogenous synthesis or maternal-fetal transport?

    NARCIS (Netherlands)

    Baardman, Maria E.; Erwich, Jan Jaap H. M.; Berger, Rolf M. F.; Hofstra, Robert M. W.; Kerstjens-Frederikse, Wilhelmina S.; Luetjohann, Dieter; Plosch, Torsten; Lutjohann, D.

    OBJECTIVE: Cholesterol is crucial for fetal development. To gain more insight into the origin of the fetal cholesterol pool in early human pregnancy, we determined cholesterol and its precursors in the amniotic fluid of uncomplicated, singleton human pregnancies. STUDY DESIGN: Total sterols were

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

    Science.gov (United States)

    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.

  13. [About the estimation of fetal risk by evalution of the serum -- amniotic liqour -- ratio of HPL during the pregnancy (author's transl)].

    Science.gov (United States)

    Altmann, P; Kucera, H; Spona, J

    1975-08-01

    Radioimmunoassay of human placental lactogen (HPL) is a valuable diagnostic tool for the early recognition of placental dysfunction. But, the rather wide normal range hampers HPL estimations to be used as a good prognostic tool. However, the simultaneous determination of HPL in maternal serum and in amniotic fluid allows a better diagnosis of the fetal well-being. Results of amniotic fluid HPL levels are expressed as percent of maternal serum levels. In this pilot study it was shown that this ratio of maternal serum HPL level to amniotic fluid level is below 10% in normal pregnancies. The ratio is between 10 to 20% in subjects where fetal distress was recorded, and dystrophic babies were observed. The value is above 20% in cases where intrauterine fetal deaths were found. This good correlation of the HPL ratio in maternal serum and amniotic fluid with the outcome of pregnancy offers a new means of using HPL estimation as prognostic tool.

  14. Bronchial stump closure with amniotic membrane in animal model

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    Gholamreza Mohajeri

    2014-01-01

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

  15. 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

  16. Detection of Bacteroides fragilis endotoxin in amniotic fluid by counterimmunoelectrophoresis

    NARCIS (Netherlands)

    I. Beckmann (Ilse); K. de Graaff (K.); F. Meisel-Mikolajczyk; H.C.S. Wallenburg (Henk)

    1994-01-01

    textabstractThe ability of counter immunoelectrophoresis (CIE) to detectBacteroides fragilis endotoxin in amniotic fluid in small concentrations was evaluated. A method was developed which, in combination with ultrafiltration, permits detection ofB. fragilis endotoxin in amniotic fluid in a

  17. Meconium-stained amniotic fluid – what is the evidence ...

    African Journals Online (AJOL)

    Meconium is rarely passed into the amniotic fluid before 34 weeks although it is present in the fetal gut from as early as 10 weeks. The passage of meconium into the amniotic fluid is a feature of advancing gestational age and reflects physiological maturation of the fetal gut. Certain specific maternal and fetal disorders may ...

  18. 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 ...

  19. HIV-1 RNA detection in the amniotic fluid of HIV-infected pregnant women

    Directory of Open Access Journals (Sweden)

    Ana Christina Lobato

    2010-08-01

    Full Text Available This study is aimed at evaluating the potential to detect human immunodeficiency virus (HIV in amniotic fluid (AF collected at delivery from 40 HIV-positive pregnant women. Thirty patients had a plasma viral load (VL below 1,000 copies/mL at delivery. VL was positive in three AF samples. No significant association was found between the HIV-1 RNA in AF and the maternal plasma samples. There was no HIV vertical transmission detected.

  20. 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).

  1. Method for chronic catheterisation of the amniotic sac in cattle.

    Science.gov (United States)

    Mohamed, A R; Noakes, D E

    1983-07-01

    The chronic catheterisation of the amniotic sac of eight pregnant cows of 180 to 200 days gestational age is described using an indwelling catheter of polyvinyl tubing. Although strict asepsis was followed, the main problem encountered was preventing contaminant bacteria from overwhelming the fetus, causing its death. This was successfully achieved in four animals, three of which received a mixture of ampicillin and cloxacillin sodium intra-amniotically and parenterally in the dam. Daily sequential samples of amniotic fluid were readily obtained until just before fetal death.

  2. 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.

  3. Amniotic Tissues for the Treatment of Chronic Plantar Fasciosis and Achilles Tendinosis

    Directory of Open Access Journals (Sweden)

    Bruce Werber

    2015-01-01

    Full Text Available Introduction. Allogeneic amniotic tissue and fluid may be used to treat chronic plantar fasciosis and Achilles tendinosis. This innovative approach involves delivering a unique allograft of live human cells in a nonimmunogenic structural tissue matrix to treat chronic tendon injury. These tissues convey very positive regenerative attributes; procurement is performed with maternal consent during elective caesarian birth. Materials and Methods. In the present investigation all patients were unresponsive to multiple standard therapies for a minimum of 6 months and were treated with one implantation of PalinGen SportFLOW around the plantar fascia and/or around the Achilles paratenon. The patients were given a standard protocol for postimplant active rehabilitation. Results. The analogue pretreatment pain score (VAS of 8. By the fourth week after treatment, all patients had significantly reduced self-reported pain. Twelve weeks following the procedure the average pain level had reduced to only 2. No adverse reactions were reported in any of the patients. Conclusion. All patients in this study experienced heel or Achilles pain, unresponsive to standard therapy protocols. After treatment all patients noted significant pain reduction, indicating that granulized amniotic membrane and amniotic fluid can be successfully used to treat both chronic plantar fasciosis and Achilles tendinosis.

  4. Amniotic band and foot polydactyly: A rare case

    Directory of Open Access Journals (Sweden)

    Mustafa Kursat Evrenos

    2016-04-01

    Full Text Available Amniotic band syndrome (ABS is defined as the triad of fetal deformation, malformation, and amputation caused by adhesions and entanglement of the fetal parts by the placenta, lacking an amniotic membrane and the remnants of amniotic membrane. The current case report presents a rare case with amniotic band syndrome, in whom constriction bands in the left hand second finger and left foot second and fourth toes are accompanied by preaxial polydactyly in the left foot. ABS is a rare condition observed in infants and it is frequently accompanied by other extremity anomalies. Appropriate treatment options allow better functional and cosmetic outcomes. [Hand Microsurg 2016; 5(1.000: 32-33

  5. 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.

  6. 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 (pspina bifida (n=100), there were higher percentages of defect coverage (either partial or complete) in both afMSC and pMSC groups compared with saline and untreated groups (pspina bifida after concentrated intraamniotic injection in the rodent model. This broadens the options for timing and cell source for TRASCET as a potential alternative in the prenatal management of spina bifida. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Amniotic fluid phthalate levels and male fetal gonad function.

    Science.gov (United States)

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

    2015-01-01

    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. 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. Mono(2-ethyl-5-carboxypentyl) phthalate (5cx-MEPP, DEHP metabolite) was not consistently associated with cryptorchidism or hypospadias. However, we observed an 18% higher (95% confidence interval [CI] = 5%-33%) testosterone level, and a 41% lower (-56% to -21%) insulin-like factor 3 level in the highest 5cx-MEPP tertile compared with the lowest. Mono(4-methyl-7-carboxyheptyl) phthalate (7cx-MMeHP, DiNP metabolite) showed elevated odds ratio point estimates for having cryptorchidism (odds ratio = 1.28 [95% CI = 0.80 to 2.01]) and hypospadias (1.69 [0.78 to 3.67]), but was not consistently associated with the steroid hormones or insulin-like factor 3. Data on the DEHP metabolite indicate possible interference with human male fetal gonadal function. Considering the DiNP metabolite, we cannot exclude (nor statistically confirm) an association with hypospadias and, less strongly, with cryptorchidism.

  8. 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

  9. Amniotic fluid 17-hydroxyprogesterone in early pregnancy.

    Science.gov (United States)

    Sulcová, J; Macek, M; Chudoba, D; Hubl, W; Hill, M; Stárka, L

    1994-12-01

    The results of measurement of 17-hydroxyprogesterone (17-OH-P) in 125 samples of amniotic fluid (AF) from early amniocenteses are presented. The fetuses from all pregnancies studied were unaffected by congenital adrenal hyperphasia caused by 21-hydroxylase deficiency. The AF 17-OH-P level increases slightly but significantly between the 11th and 15th week of gestation, with a maximum in the 14th week. There is no difference between the values measured in male and female fetuses. The AF 17-OH-P levels from the early gestation were compared with those from the 16th-22nd week of pregnancy (published previously). The overall differences of AF 17-OH-P concentrations when considered in all gestational age groups in the whole period 12-22 weeks were statistically insignificant. Thus, the biochemical prenatal diagnosis of congenital adrenal hyperplasia due to 21-hydroxylase deficiency and control of its early fetal treatment could be carried out starting from the end of the first trimester in the same way as at the later period of gestation.

  10. Abnormal ultrasound appearance of the amniotic membranes - diagnostic and significance: a pictorial essay.

    Science.gov (United States)

    Pleș, Liana; Sima, Romina Marina; Moisei, Cristina; Moga, Marius Alexandru; Dracea, Laura

    2017-04-22

    The obstetrical ultrasound may identify the protrusion of the uterine surface or placental area into the amniotic cavity. The differential diagnosis of this pathology with uterine adhesions, septate uterus, circumvallate placenta, amniotic band or amniotic sheet can be sometimes difficult. The purpose of the pictorial essay is to exemplify the presence of the amniotic sheet and circumvallate placenta in routine obstetrics screening of all trimesters of pregnancy.

  11. Management of Amniotic Sheet with a Hammock-like Placenta

    Directory of Open Access Journals (Sweden)

    Liangcheng Wang

    2016-09-01

    Full Text Available An amniotic sheet is a septation in the amniotic cavity with a perforation that allows amniotic fluid to pass through. Although the incidence of abnormal placental implantation is higher in such cases, the management recommendations remain unclear. We report a case of an amniotic sheet with a hammock-like placenta located in the center of the uterine cavity. A 25-year-old woman with a history of two dilation and curettage procedures was found to have an amniotic cavity separated by a septum that contained part of the placenta. At gestational Week 32, magnetic resonance images revealed that the placenta was attached from the anterior to posterior uterine walls and resembled a hammock hanging in the center of the uterus. Subsequently, continuous intravenous administration of ritodrine hydrochloride and magnesium sulfate were given. The pregnancy was extended to Week 36. Elective cesarean section was performed, and a 3212-g female infant was delivered. Thus, owing to the risk of umbilical cord complications and placental injury secondary to premature rupture of membranes, aggressive and careful perinatal management is required in such cases.

  12. 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

  13. 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.

  14. 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

  15. The relationship between amniotic fluid matrix metalloproteinase-8 and funisitis.

    Science.gov (United States)

    Park, J S; Romero, R; Yoon, B H; Moon, J B; Oh, S Y; Han, S Y; Ko, E M

    2001-11-01

    The fetal inflammatory response syndrome is a multisystem disorder associated with impending preterm delivery and adverse neonatal outcome. Inflammation of the umbilical cord--funisitis--is the histologic counterpart of fetal inflammatory response syndrome and has been associated with an increased risk for the development of cerebral palsy. Neutrophils found in the amniotic cavity are of fetal origin. Therefore, neutrophil secretory products may be an index of the fetal inflammatory response syndrome. To test this hypothesis, we examined the relationship between levels of amniotic fluid matrix metalloproteinase-8 and funisitis. The relationship between the presence of funisitis and concentrations of amniotic fluid matrix metalloproteinase-8 was examined in 255 consecutive patients who delivered preterm singleton neonates (gestational age, jelly. Matrix metalloproteinase-8 was measured by use of a specific immunoassay. Nonparametric statistics were used for analysis. Funisitis was present in 23% (59/255) of cases. Patients with funisitis had a significantly higher median concentration of amniotic fluid matrix metalloproteinase-8 than those without funisitis (median, 433.7 ng/mL [range, 1.5-3836.8 ng/mL] vs median, 1.9 ng/mL [range, <0.3-4202.7 ng/mL]; P <.001). The diagnostic indices of matrix metalloproteinase-8 (cutoff, 23 ng/mL) in the identification of funisitis were: sensitivity of 90% (53/59), specificity of 78% (153/196), positive predictive value of 55% (53/96), and negative predictive value of 96% (153/159). There is a strong association between increased levels of amniotic fluid matrix metalloproteinase-8 and funisitis. We propose that determination of amniotic fluid matrix metalloproteinase-8 concentrations may assist the assessment of the fetal inflammatory status, thereby eliminating the need for fetal blood sampling.

  16. Amniotic membrane transplantation for acute ocular burns.

    Science.gov (United States)

    Clare, Gerry; Suleman, Hanif; Bunce, Catey; Dua, Harminder

    2012-09-12

    Ocular surface burns can be caused by chemicals (alkalis and acids) or by direct heat. Amniotic membrane transplantation (AMT) performed in the acute phase (day 0 to day 7) of an ocular surface burn is reported to relieve pain, accelerate healing and reduce scarring and blood vessel formation. The surgery involves applying a patch of amniotic membrane (AM) over the entire ocular surface up to the eyelid margins. To assess the effects of AMT on the eyes of people having suffered acute ocular surface burns. We searched CENTRAL (which contains the Cochrane Eyes and Vision Group Trials Register) (The Cochrane Library 2012, Issue 6), MEDLINE (January 1946 to June 2012), EMBASE (January 1980 to June 2012), Latin American and Caribbean Literature on Health Sciences (LILACS) (January 1982 to June 2012), the metaRegister of Controlled Trials (mRCT) (www.controlled-trials.com), ClinicalTrials.gov (www.clinicaltrials.gov) and the WHO International Clinical Trials Registry Platform (ICTRP) (www.who.int/ictrp/search/en). We did not use any date or language restrictions in the electronic searches for trials. We last searched the electronic databases on 11 June 2012. We included randomised trials of medical therapy and AMT applied in the first seven days after an ocular surface burn compared to medical therapy alone. Two authors independently assessed the risk of bias of included studies and extracted relevant data. We contacted trial investigators for missing information. We summarised data using risk ratios (RRs) and mean differences (MDs) as appropriate. We included one RCT of 100 participants with ocular burns that were randomised to treatment with AMT and medical therapy or medical therapy alone. A subset of patients (n = 68) who were treated within the first seven days of the injury met the inclusion criteria and were included in the analysis. The remaining 32 eyes were excluded. The included subset consisted of 36 moderate (Dua classification II-III) and 32 severe (Dua

  17. Procoagulant extracellular vesicles in amniotic fluid.

    Science.gov (United States)

    Hell, Lena; Wisgrill, Lukas; Ay, Cihan; Spittler, Andreas; Schwameis, Michael; Jilma, Bernd; Pabinger, Ingrid; Altevogt, Peter; Thaler, Johannes

    2017-06-01

    Embolization of amniotic fluid (AF) into the blood circulation leads to disseminated intravascular coagulation (DIC). Procoagulant phosphatidylserine (PS)- and tissue factor (TF)-exposing extracellular vesicles (EVs) might play an important role in AF embolism-induced DIC. It was the aim of the present study to perform analyses of the procoagulant properties of AF with a panel of functional coagulation assays and flow cytometry. We applied a prothrombinase assay (that quantifies PS exposure on EVs), an EV-associated TF activity assay, a fibrin generation assay, a thrombin generation assay, a whole blood clotting model, and flow cytometry in AF and control plasma. We found that PS exposure on EVs was 21-fold increased in AF compared with plasma. Also, EV-associated TF activity was highly increased in AF compared with plasma. AF-derived EVs activated the blood coagulation cascade via PS and TF in the fibrin and thrombin generation assays. In a whole blood clotting model, AF-derived EVs significantly shortened the clotting time from 734 ± 139 seconds in the presence to 232 ± 139 seconds in the absence of an anti-TF antibody. The contact activation pathway via factor XII (FXII) was not affected. Applying flow cytometry, a subpopulation of PS+ and TF+ EVs was identified in AF but not in control plasma. In conclusion, we investigated the effect of AF on blood coagulation and found that PS+ and TF+ EVs determine their procoagulant potential. Taken together, our data further delineate the pathomechanisms underlying AF-induced coagulopathy. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. 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.

  19. A novel method to derive amniotic fluid stem cells for therapeutic purposes

    Directory of Open Access Journals (Sweden)

    Vantanasiri Chanchai

    2010-10-01

    Full Text Available Abstract Background Human amniotic fluid stem (hAFS cells have become an attractive stem cell source for medical therapy due to both their ability to propagate as stem cells and the lack of ethical debate that comes with the use of embryonic stem cells. Although techniques to derive stem cells from amniotic fluid are available, the techniques have limitations for clinical uses, including a requirement of long periods of time for stem cell production, population heterogeneity and xeno-contamination from using animal antibody-coated magnetic beads. Herein we describe a novel isolation method that fits for hAFS derivation for cell-based therapy. Methods and Results With our method, single hAFS cells generate colonies in a primary culture of amniotic fluid cells. Individual hAFS colonies are then expanded by subculturing in order to make a clonal hAFS cell line. This method allows derivation of a substantial amount of a pure stem cell population within a short period of time. Indeed, 108 cells from a clonal hAFS line can be derived in two weeks using our method, while previous techniques require two months. The resultant hAFS cells show a 2-5 times greater proliferative ability than with previous techniques and a population doubling time of 0.8 days. The hAFS cells exhibit typical hAFS cell characteristics including the ability to differentiate into adipogenic-, osteogenic- and neurogenic lineages, expression of specific stem cell markers including Oct4, SSEA4, CD29, CD44, CD73, CD90, CD105 and CD133, and maintenance of a normal karyotype over long culture periods. Conclusions We have created a novel hAFS cell derivation method that can produce a vast amount of high quality stem cells within a short period of time. Our technique makes possibility for providing autogenic fetal stem cells and allogeneic cells for future cell-based therapy.

  20. The evolution of amniote gastrulation: the blastopore-primitive streak transition.

    Science.gov (United States)

    Stower, Matthew J; Bertocchini, Federica

    2017-03-01

    In the animal kingdom, gastrulation, the process by which the primary germ layers are formed involves a dramatic transformation in the topology of the cells that give rise to all of the tissues of the adult. Initially formed as a mono-layer, this tissue, the epiblast, becomes subdivided through the internalization of cells, thereby forming a two (bi-laminar) or three (tri-laminar) layered embryo. This morphogenetic process coordinates the development of the fundamental body plan and the three-body axes (antero-posterior, dorso-ventral, and left-right) and begins a fundamental segregation of cells toward divergent developmental fates. In humans and other mammals, as well as in avians, gastrulating cells internalize along a structure, called the primitive streak, which builds from the periphery toward the center of the embryo. How these morphogenetic movements are orchestrated and evolved has been a question for developmental biologists for many years. Is the primitive streak a feature shared by the whole amniote clade? Insights from reptiles suggest that the primitive streak arose independently in mammals and avians, while the reptilian internalization site is a structure half-way between an amphibian blastopore and a primitive streak. The molecular machinery driving primitive streak formation has been partially dissected using mainly the avian embryo, revealing a paramount role of the planar cell polarity (PCP) pathway in streak formation. How did the employment of this machinery evolve? The reptilian branch of the amniote clade might provide us with useful tools to investigate the evolution of the amniote internalization site up to the formation of the primitive streak. WIREs Dev Biol 2017, 6:e262. doi: 10.1002/wdev.262 For further resources related to this article, please visit the WIREs website. © 2017 Wiley Periodicals, Inc.

  1. Inference of the protokaryotypes of amniotes and tetrapods and the evolutionary processes of microchromosomes from comparative gene mapping.

    Directory of Open Access Journals (Sweden)

    Yoshinobu Uno

    Full Text Available Comparative genome analysis of non-avian reptiles and amphibians provides important clues about the process of genome evolution in tetrapods. However, there is still only limited information available on the genome structures of these organisms. Consequently, the protokaryotypes of amniotes and tetrapods and the evolutionary processes of microchromosomes in tetrapods remain poorly understood. We constructed chromosome maps of functional genes for the Chinese soft-shelled turtle (Pelodiscus sinensis, the Siamese crocodile (Crocodylus siamensis, and the Western clawed frog (Xenopus tropicalis and compared them with genome and/or chromosome maps of other tetrapod species (salamander, lizard, snake, chicken, and human. This is the first report on the protokaryotypes of amniotes and tetrapods and the evolutionary processes of microchromosomes inferred from comparative genomic analysis of vertebrates, which cover all major non-avian reptilian taxa (Squamata, Crocodilia, Testudines. The eight largest macrochromosomes of the turtle and chicken were equivalent, and 11 linkage groups had also remained intact in the crocodile. Linkage groups of the chicken macrochromosomes were also highly conserved in X. tropicalis, two squamates, and the salamander, but not in human. Chicken microchromosomal linkages were conserved in the squamates, which have fewer microchromosomes than chicken, and also in Xenopus and the salamander, which both lack microchromosomes; in the latter, the chicken microchromosomal segments have been integrated into macrochromosomes. Our present findings open up the possibility that the ancestral amniotes and tetrapods had at least 10 large genetic linkage groups and many microchromosomes, which corresponded to the chicken macro- and microchromosomes, respectively. The turtle and chicken might retain the microchromosomes of the amniote protokaryotype almost intact. The decrease in number and/or disappearance of microchromosomes by repeated

  2. Mouse embryos cultured in amniotic fluid | Oettle | South African ...

    African Journals Online (AJOL)

    One thousand mouse embryos were collected over a period of 6 weeks·and randomly assigned to either amniotic fluid aspirated during routine amniocentesis from normal 16 weeks pregnant patients or Earle's medium. The embryos were cultured for 72 hours at 37°C in 5% carbon dioxide in air. Osmolarity, pH, partial ...

  3. Elemental Profile in Amniotic Fluid of some Nigerian Pregnant Women

    African Journals Online (AJOL)

    In this study concentration level of calcium, cadmium, copper, iron, magnesium, manganese, nickel, lead and zinc were determined in the amniotic fluid of pregnant women, aged 15 – 45 years enrolled at the Obafemi Awolowo University Teaching Hospitals Complex Ile – Ife. This was with a view to predict the body burden ...

  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. Short term results of pterygium surgery with adjunctive amniotic ...

    African Journals Online (AJOL)

    2012-11-30

    Nov 30, 2012 ... Short term results of pterygium surgery with adjunctive amniotic membrane graft. O Okoye, NC Oguego, CM Chuka Okosa, M Ghanta1. Department of Ophthalmology, University of Nigeria Teaching Hospital, Ituku Ozalla, Enugu Nigeria,. 1Goutani Eye Institute, Rajahmundry, Andra Pradesh, India.

  6. 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.

  7. Assessment of DNA Damage by RAPD in Paracentrotus lividus Embryos Exposed to Amniotic Fluid from Residents Living Close to Waste Landfill Sites

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    Maurizio Guida

    2010-01-01

    Full Text Available The aim of this study was to assess the genotoxic effects of environmental chemicals on residents living near landfills. The study was based on samples of amniotic fluid from women living in the intensely polluted areas around the Campania region of Italy compared to a nonexposed control group. We evaluated the genetic effects that this amniotic fluids collected in contaminated sites had on Paracentrotus lividus embryos. DNA damage was detected through changes in RAPD (Random Amplified Polymorphism DNA profiles. The absence of the amplified DNA fragments indicated deletions in Paracentrotus lividus DNA exposed to the contaminated amniotic fluids when compared to equal exposure to uncontaminated fluids. These results show the ability of RAPD-PCR to detect and isolate DNA sequences representing genetic alterations induced in P. lividus embryos. Using this method, we identified two candidate target regions for DNA alterations in the genome of P. lividus. Our research indicates that RAPD-PCR in P. lividus embryo DNA can provide a molecular approach for studying DNA damage from pollutants that can impact human health. To our knowledge, this is the first time that assessment of DNA damage in P. lividus embryos has been tested using the RAPD strategy after exposure to amniotic fluid from residents near waste landfill sites.

  8. Sealing of Corneal Lacerations Using Photo-Activated Rose Bengal Dye and Amniotic Membrane

    Science.gov (United States)

    2017-05-05

    Activated Rose Bengal Dye and Amniotic Membrane presented at/published to Sun and Science, Galveston, Texas, 22 April 2017 & American Society of...Corneal Lacerations Using Photo-Activated Rose Bengal Dye and Amniotic Membrane 1. TITLE OF MATERIAL TO BE PUBLISHED OR PRESENTED Sealing of...Corneal Lacerations Using Photo-Activated Rose Bengal Dye and Amniotic Membrane 2. FUNDING RECEIVED FOR THIS STUDY? [gl YES D NO FUNDING SOURCE: jus

  9. Transplante de membrana amniótica Amniotic membrane transplantation

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    Hamilton Moreira

    2000-08-01

    Full Text Available O transplante de membrana amniótica tem sido utilizado como alternativa para a reconstrução da superfície ocular em substituição ao tecido conjuntival nos casos de doenças cicatriciais da córnea ou conjuntiva. Tem sido descrito na literatura para o tratamento de defeitos epiteliais persistentes, pterígio recidivado, Síndrome de Stevens-Johnson e Penfigóide cicatricial, queimaduras químicas e ceratopatia bolhosa.Amniotic membrane transplantation has been used as an alternative for ocular surface reconstruction. Indications for amniotic membrane transplantation include persistent epithelial defects, pterygium, Stevens-Johnson syndrome, ocular cicatricial pemphigoid, chemical burns and pseudo-phakik bullous keratopathy.

  10. The oldest record of aquatic amniote congenital scoliosis.

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    Tomasz Szczygielski

    Full Text Available We report the first occurrence of congenital scoliosis in an early Permian aquatic parareptile, Stereosternum tumidum from Paraná state, Brazil. The spine malformation is caused by a congenital hemivertebra. These observations give insight into the biomechanical aspects of underwater locomotion in an axial skeleton-compromised aquatic amniote. This is the oldest record of a hemivertebra in an aquatic animal.

  11. Amniotic Fluid Infection in Preterm Pregnancies with Intact Membranes

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    Tarja Myntti

    2017-01-01

    Full Text Available Introduction. Intra-amniotic infection (IAI is a major cause of preterm labor and adverse neonatal outcome. We evaluated amniotic fluid (AF proteolytic cascade forming biomarkers in relation to microbial invasion of the amniotic cavity (MIAC and IAI in preterm pregnancies with intact membranes. Material and Methods. Amniocentesis was made to 73 women with singleton pregnancies; 27 with suspected IAI; and 46 controls. AF biomarkers were divided into three cascades: Cascade 1: matrix metalloproteinase-8 (MMP-8, MMP-9, myeloperoxidase (MPO, and interleukin-6; Cascade 2: neutrophil elastase (HNE, elafin, and MMP-9; Cascade 3: MMP-2, tissue inhibitor of matrix metalloproteinases-1 (TIMP-1, MMP-8/TIMP-1 molar ratio, and C-reactive protein (CRP. MMP-8 was measured by an immunoenzymometric assay and the others were measured by ELISA. Standard biochemical methods, molecular microbiology, and culture techniques were used. Results. MMP-8, MMP-9, MPO, elafin, and TIMP-1 concentrations were higher in IAI suspected cases compared to controls and also in IAI suspected cases with MIAC compared to those without MIAC when adjusted by gestational age at amniocentesis. All biomarkers except elafin and MMP-2 had the sensitivity of 100% with thresholds based on ROC-curve. Odd ratios of biomarkers for MIAC were 1.2-38 and 95% confidential intervals 1.0-353.6. Conclusions. Neutrophil based AF biomarkers were associated with IAI and MIAC.

  12. Adverse perinatal outcomes in borderline amniotic fluid index.

    Science.gov (United States)

    Jamal, Ashraf; Kazemi, Maryam; Marsoosi, Vajiheh; Eslamian, Laleh

    2016-11-01

    Normal amniotic fluid predicts normal placental function, fetal growth and fetal well-being. To determine adverse pregnancy outcomes in borderline amniotic fluid index (AFI). Pregnant women (37-40 wks) with diagnosis of borderline AFI between December 2012 and August 2014 were identified. Antepartum, intrapartum and neonatal data were collected and compared with those of pregnant women with normal AFI. An AFI less than 8 and more than 5 cm was defined for borderline AFI. Pregnancy outcomes included Cesarean section for non-reassuring fetal heart rate, meconium stained amniotic fluid, 5-min Apgar score borderline AFI was significantly lower than normal AFI. Cesarean section rate for non-reassuring fetal heart rate in women of borderline AFI was significantly higher and there was an increased incidence of birth weight less than 10(th) percentile for gestation age in borderline AFI group. Incidence of low Apgar score and low umbilical artery pH in pregnancies with borderline AFI was significantly higher than women with normal AFI. There were no significant difference in the rate of NICU admission and meconium staining in both groups. There are significant differences for adverse pregnancy outcomes , such as Cesarean section due to non-reassuring fetal heart rate, birth weight less than 10(th) percentile for gestation age, low 5 min Apgar score and low umbilical artery pH between pregnancies with borderline and normal AFI.

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

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    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.

  14. The evolution of epigenetic regulators CTCF and BORIS/CTCFL in amniotes.

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    Timothy A Hore

    2008-08-01

    Full Text Available CTCF is an essential, ubiquitously expressed DNA-binding protein responsible for insulator function, nuclear architecture, and transcriptional control within vertebrates. The gene CTCF was proposed to have duplicated in early mammals, giving rise to a paralogue called "brother of regulator of imprinted sites" (BORIS or CTCFL with DNA binding capabilities similar to CTCF, but testis-specific expression in humans and mice. CTCF and BORIS have opposite regulatory effects on human cancer-testis genes, the anti-apoptotic BAG1 gene, the insulin-like growth factor 2/H19 imprint control region (IGF2/H19 ICR, and show mutually exclusive expression in humans and mice, suggesting that they are antagonistic epigenetic regulators. We discovered orthologues of BORIS in at least two reptilian species and found traces of its sequence in the chicken genome, implying that the duplication giving rise to BORIS occurred much earlier than previously thought. We analysed the expression of CTCF and BORIS in a range of amniotes by conventional and quantitative PCR. BORIS, as well as CTCF, was found widely expressed in monotremes (platypus and reptiles (bearded dragon, suggesting redundancy or cooperation between these genes in a common amniote ancestor. However, we discovered that BORIS expression was gonad-specific in marsupials (tammar wallaby and eutherians (cattle, implying that a functional change occurred in BORIS during the early evolution of therian mammals. Since therians show imprinting of IGF2 but other vertebrate taxa do not, we speculate that CTCF and BORIS evolved specialised functions along with the evolution of imprinting at this and other loci, coinciding with the restriction of BORIS expression to the germline and potential antagonism with CTCF.

  15. Amniotic membrane welded to contact lens by 1470-nm diode laser:a novel method for sutureless amniotic membrane transplantation

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    Rifat Rasier

    2014-12-01

    Full Text Available AIM: To avoid the side effects of the suture usage by welding amniotic membrane (AM to contact lens (CL with laser.METHODS:AM was taken from pregnant women and cleaned from blood clots with sterile phosphate-buffered physiological saline solution which included antibiotics. Stromal side of the AM was spread inside of the CL and it was welded to CL by 1470 nm diode laser. 600 μm diameter fiber tip of the laser was contacted with the epithelial side of the AM from 4 separate points. After welding excess amniotic membrane around the CL was cut with a scalpel.RESULTS:Stromal side of the AM was spread inside of the CL and then with laser fiber, different power levels and exposure times were applied on the epithelium of AM and 340 mW for seven seconds was found optimal. CL and AM attached with the spot welding effect in 4 points by touching fiber tip. CL-AM welded complex did not separated from each other while holding AM that extend beyond the CL with the help of two forceps.CONCLUSION:As a conclusion, it was aimed in this study to achieve the success of the conventional amniotic membrane transplantation (AMTwith the easiness of applying a CL and to avoid risks and side effects of corneal or conjunctival suturing. The results showed that the application of the CL–AM complex will be as easy as the application of a CL and lasts shortly.

  16. Endothelial properties of third-trimester amniotic fluid stem cells cultured in hypoxia.

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    Schiavo, Andrea Alex; Franzin, Chiara; Albiero, Mattia; Piccoli, Martina; Spiro, Giovanna; Bertin, Enrica; Urbani, Luca; Visentin, Silvia; Cosmi, Erich; Fadini, Gian Paolo; De Coppi, Paolo; Pozzobon, Michela

    2015-10-31

    Endothelial dysfunction is found in different pathologies such as diabetes and renal and heart diseases, representing one of the major health problems. The reduced vasodilation of impaired endothelium starts a prothrombotic state associated with irregular blood flow. We aimed to explore the potential of amniotic fluid stem (AFS) cells as a source for regenerative medicine in this field; for the first time, we focused on third-trimester amniotic fluid AFS cells and compared them with the already-described AFS cells from the second trimester. Cells from the two trimesters were cultured, selected and expanded in normoxia (20 % oxygen) and hypoxia (5 % oxygen). Cells were analysed to compare markers, proliferation rate and differentiation abilities. Endothelial potential was assessed not only in vitro-Matrigel tube formation assay, acetylated human low-density lipoprotein (AcLDL) uptake-but also in vivo (Matrigel plug with cell injection and two animal models). Specifically, for the latter, we used established protocols to assess the involvement of AFS cells in two different mouse models of endothelial dysfunction: (1) a chronic ischemia model with local injection of cells and (2) an electric carotid damage where cells were systemically injected. We isolated and expanded AFS cells from third-trimester amniotic fluid samples by using CD117 as a selection marker. Hypoxia enhanced the proliferation rate, the surface protein pattern was conserved between the trimesters and comparable differentiation was achieved after culture in both normoxia and hypoxia. Notably, the expression of early endothelial transcription factors and AngiomiRs was detected before and after induction. When in vivo, AFS cells from both trimesters expanded in hypoxia were able to rescue the surface blood flow when locally injected in mice after chronic ischemia damage, and importantly AFS cells at term of gestation possessed enhanced ability to fix carotid artery electric damage compared with AFS

  17. Severe maternal morbidity and mortality from amniotic fluid embolism in the Netherlands

    NARCIS (Netherlands)

    Stolk, K.H.; Zwart, J.J.; Schutte, J.; van Roosmalen, J.

    2012-01-01

    We have assessed the incidence, symptoms and risk factors of amniotic fluid embolism in the Netherlands. Data were retrieved from two nationwide registration systems. From 1983 to 2005 the maternal mortality ratio of amniotic fluid embolism increased from 0.11 to 0.63 (odds ratio (OR) 5.8, 95%

  18. 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

  19. 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....

  20. Functional and proteomic analysis of serum and cerebrospinal fluid derived from patients with traumatic brain injury: a pilot study.

    Science.gov (United States)

    Cadosch, Dieter; Thyer, Matthew; Gautschi, Oliver P; Lochnit, Günter; Frey, Sönke P; Zellweger, René; Filgueira, Luis; Skirving, Allan P

    2010-01-01

    An enhanced fracture healing response has been reported in patients with traumatic brain injury (TBI). This has been attributed to circulating humoral factors that are thought to be proteins produced and released by the injured brain. However, these factors remain unknown. The aim of this study was to identify osteogenic factors in serum and cerebrospinal fluid (CSF) from TBI patients. This was carried out using in vitro proliferation assays with the human foetal osteoblastic 1.19 cell line (hFOB) combined with a novel proteomic approach. Serum was collected from brain-injured (n = 12) and non-brain-injured (n = 9) patients with a comorbid femur shaft fracture. Similarly, CSF was obtained from TBI (n = 7) and non-TBI (n = 9) patients. The osteoinductive potential of these samples was determined by measuring the in vitro proliferation rate of hFOB cells. Highly osteogenic serum and CSF samples of TBI patients were chosen for protein analysis and were compared to those of non-brain-injured patients. A new hFOB cell-based method was used to enrich the proteins in these samples, which had a functional affinity for these osteoprogenitor cells. These enriched protein fractions were mapped using two-dimensional gel electrophoresis and protein imaging methods displaying serum and CSF proteins of brain-injured and control subjects that had an affinity for human osteoprogenitor cells. Serum and CSF derived from brain-injured patients demonstrated a greater osteoinductive potential (P < 0.05) than their non-brain-injured counterparts. Clear-cut differences in the pattern of proteins in two-dimensional gels were detected between TBI and control patients. Fourteen proteins were exclusively present in the serum of TBI patients, while other proteins were either up- or downregulated in samples collected from TBI patients (P < 0.05). Osteoinductive factors are present in the serum and CSF of brain-injured patients. These may include one or more of those proteins identified as

  1. Construction of tissue-engineered cornea composed of amniotic epithelial cells and acellular porcine cornea for treating corneal alkali burn.

    Science.gov (United States)

    Luo, Hailang; Lu, Yongbo; Wu, Tiantian; Zhang, Mi; Zhang, Yongjie; Jin, Yan

    2013-09-01

    Although acellular corneas have been reported to be a potential substitute for allogeneic cornea transplantation to treat corneal injury, severe corneal injury is hard to repair due to inflammation and neovascularization. The use of the amniotic membrane as a graft in ocular surface reconstruction has become widespread because of the anti-inflammatory and anti-angiogenic properties of amniotic epithelial cells (AECs). Our objective was to construct a tissue-engineered cornea (TEC) composed of an acellular porcine cornea (APC) and AECs to repair severe corneal injury. Corneal cells were completely removed from the prepared APC, and the microstructure, mechanical properties, and stability of a natural porcine cornea (NPC) was maintained. In vitro, MTT and flow cytometry analyses showed that the APC did not negatively affect cell viability and apoptosis. In vivo, corneal pocket and subcutaneous transplantation demonstrated that the APC was incapable of trigging accepted immune response. AECs isolated from the human amniotic membrane have proliferation potential and present healthy morphology before 6 passages. After 7 days of culture on the surface of the APC, the AECs were stratified into 5-6 layers. We found that the AECs reconstituted the basement membrane that had been disrupted by the decellularization process. ELISA results showed that after culturing the TEC, the culture medium contained anti-inflammatory and anti-angiogenic growth factors, such as MIF, IL6, Fas-L, and PDEF. Finally, the results of lamellar keratoplasty to treat an alkali burn showed that the transplanted TEC was transparent and completely inoculated into the host cornea. However, the transplanted APC was degraded due to host rejection. Therefore, we conclude that a TEC composed of AECs and an APC holds great potential for the repair of severe corneal injury. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. 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.

  3. [Amniotic membrane transplantation in severe corneal epithelial diseases. Preliminary results].

    Science.gov (United States)

    Zito, E; Borderie, V; Touzeau, O; Bourcier, T; Allouch, C; Laroche, L

    2002-11-01

    To evaluate amniotic membrane transplantation (AMT) in severe corneal epithelial diseases. Amniotic membrane transplantation was performed in 14 eyes of 14 patients from four groups: A, five severe ocular burns; B, four cases of cicatricial keratoconjunctivitis; C, three persistent epithelial defects after penetrating keratoplasty; D, two cases of pseudophakic bullous keratopathy. Five patients underwent AMT alone; two patients underwent AMT combined with limbal transplantation; the other three patients had limbal transplantation performed before AMT. Eight patients required combined penetrating keratoplasty. Patients with corneal stable reepithelialization, no corneal neovascularization, and no recurrence of the initial pathology were considered successful. The mean follow-up was 7+/-3 months. All but three patients underwent corneal reepithelialization within 6 weeks of AMT, with a mean healing time of 31+/-23 days. The success rate was 75% at 6 months (Kaplan-Meier method). Three of four procedures in group B failed. In eight patients, visual acuity improved, in one it worsened, and in the last five patients it remained unchanged. Visual acuity increased by an average of 7+/-9 lines. AMT is a useful technique for ocular surface reconstruction, especially in association with limbal transplantation. It could also improve the prognosis of penetrating keratoplasty in patients with severe corneal conditions.

  4. 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)

  5. Epithelial wound healing on keratin film, amniotic membrane and polystyrene in vitro.

    Science.gov (United States)

    Feng, Yaqing; Borrelli, Maria; Meyer-Ter-Vehn, Tobias; Reichl, Stephan; Schrader, Stefan; Geerling, Gerd

    2014-06-01

    Corneal epithelial wound healing is a major issue in ocular surface (OS) reconstruction. Aim of this study was to evaluate parameters of epithelial wound healing in vitro on transparent keratin films (KFs) derived from human hair in comparison with amniotic membrane (AM) and polystyrene. The human corneal epithelial cell line (HCE-T) was expanded on KF, AM and commercially available 24-well polystyrene cell culture plates in vitro to compare cell proliferation, migration and attachment by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, scratch-wound healing and adhesion assay. Cells cultured on KF and AM at an air-liquid interface for 14 d were stained with hematoxylin and eosin for histology. The highest proliferation of HCE-T cells was observed on polystyrene at all time points (p polystyrene at all time points (p polystyrene and KF (p = 0.032 and p = 0.001). No significant difference in cell attachment was observed between KF and polystyrene (p = 0.147). Histology demonstrated that HCE-T cells cultured on KF and AM at an air-liquid interface for 14 d form a multilayered epithelium similar to normal human corneal epithelium. Transparent KFs derived from human hair support proliferation, migration, adhesion and differentiation of HCE-T cells in vitro. Therefore, it could be a promising alternative to AM for OS reconstruction.

  6. 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

  7. Calcium sensing receptor expression in ovine amniotic fluid mesenchymal stem cells and the potential role of R-568 during osteogenic differentiation.

    Directory of Open Access Journals (Sweden)

    Pamela Di Tomo

    Full Text Available Amniotic fluid-derived stem (AFS cells have been identified as a promising source for cell therapy applications in bone traumatic and degenerative damage. Calcium Sensing Receptor (CaSR, a G protein-coupled receptor able to bind calcium ions, plays a physiological role in regulating bone metabolism. It is expressed in different kinds of cells, as well as in some stem cells. The bone CaSR could potentially be targeted by allosteric modulators, in particular by agonists such as calcimimetic R-568, which may potentially be helpful for the treatment of bone disease. The aim of our study was first to investigate the presence of CaSR in ovine Amniotic Fluid Mesenchymal Stem Cells (oAFMSCs and then the potential role of calcimimetics in in vitro osteogenesis. oAFMSCs were isolated, characterized and analyzed to examine the possible presence of CaSR by western blotting and flow cytometry analysis. Once we had demonstrated CaSR expression, we worked out that 1 µM R-568 was the optimal and effective concentration by cell viability test (MTT, cell number, Alkaline Phosphatase (ALP and Alizarin Red S (ARS assays. Interestingly, we observed that basal diffuse CaSR expression in oAFMSCs increased at the membrane when cells were treated with R-568 (1 µM, potentially resulting in activation of the receptor. This was associated with significantly increased cell mineralization (ALP and ARS staining and augmented intracellular calcium and Inositol trisphosphate (IP3 levels, thus demonstrating a potential role for calcimimetics during osteogenic differentiation. Calhex-231, a CaSR allosteric inhibitor, totally reversed R-568 induced mineralization. Taken together, our results demonstrate for the first time that CaSR is expressed in oAFMSCs and that calcimimetic R-568, possibly through CaSR activation, can significantly improve the osteogenic process. Hence, our study may provide useful information on the mechanisms regulating osteogenesis in oAFMSCs, perhaps

  8. Using SELDI-TOF mass spectrometry on amniotic fluid and for clinical proteomics and theranostics in disorders of pregnancy.

    Science.gov (United States)

    Buhimschi, Irina A

    2012-01-01

    Clinical proteomics encompasses a multitude of experimental approaches, tools, and techniques based on proteomics technology which are directly aimed to accelerate and improve diagnosis and treatment of human diseases. Surface-enhanced laser desorption ionization time-of-flight (SELDI-TOF) mass spectrometry is a variant of matrix-enhanced laser desorption ionization (MALDI) that makes use of chemically-modified surfaces to reduce the complexity of biological samples prior to separation in the mass analyzer. Compared to other proteomic techniques, SELDI has several important advantages such as ability to analyze complex biological samples with minimal pre-processing, ease of handling and high throughput. Importantly, once the biomarker or combination of biomarkers with potential clinical value has been established, validation analyses can be conducted in close proximity to clinical settings which is important for establishing the utility of new diagnostics in clinical decision making and perhaps future theranostic interventions. This chapter provides protocols for experimental design and methodology aimed at (1) discovering biologically relevant biomarkers in amniotic fluid using SELDI-TOF; (2) validating the clinical utility of the biomarkers as new diagnostics; (3) translating the biomarker findings into pathophysiological phenomena to provide further insight and extend the current understanding of the disease process. Many of the principles described herein for amniotic fluid could be generalized to studies involving other types of biological samples and other clinical questions.

  9. 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

  10. Amniotic membrane graft to reconstruct divided nevi of eyelids.

    Science.gov (United States)

    Bhattacharjee, Kasturi; Singh, Manpreet; Bhattacharjee, Harsha

    2015-05-24

    Two patients with congenital divided nevi of the eyelids presented with chief symptoms of cosmetic blemish (n=2) and epiphora (n=1). All the lesions were surgically excised with histological free margins and amniotic membrane graft (AMG) was used to cover the mucosal and eyelid marginal defects. Caruncular and punctal involvement was managed with complete excision and monocanalicular stent insertion to prevent punctal stenosis. AMG provided an adequate scaffold for conjunctival regenesis and anatomic continuity was found within 6 weeks in both patients. None of them had any residual pigmentation or disease recurrence after a follow-up of more than 2.5 years. Local absence of eyelashes was the only drawback which patients masked with modern artificial eyelashes or make-up. This novel utility of AMG illustrates effective conjunctival regenesis and maintenance of mucocutaneous junction functionality. 2015 BMJ Publishing Group Ltd.

  11. 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.

  12. Fetal hyperechogenic kidney with normal amniotic fluid volume: a diagnostic dilemma.

    Science.gov (United States)

    Mashiach, R; Davidovits, M; Eisenstein, B; Kidron, D; Kovo, M; Shalev, J; Merlob, P; Vardimon, D; Efrat, Z; Meizner, I

    2005-07-01

    To determine the prognostic value of sonographically detected fetal hyperechogenic kidneys with normal amniotic fluid volume. Seven cases of hyperechogenic fetal kidneys were identified by sonography over a 7-year period (1996--2002). Increased renal echogenicity was diagnosed when the renal parenchyma was of greater echogenicity than adjacent liver tissue. Amniotic fluid volume was measured by the semiquantitative sonographic technique known as the amniotic fluid index (AFI). Three of the live-born infants had autosomal dominant polycystic kidney disease and one had autosomal recessive polycystic kidney. In the remainder, autopsy study revealed multifocal renal dysplasia in two cases and normal kidneys in one. Increased renal echogenicity with normal amniotic fluid volume in a fetus without other anomalies is a difficult diagnostic dilemma. Although it is usually indicative of renal parenchymal disease with possible renal failure after birth or in early childhood, in some cases, it represents a normal variant. . Copyright (c) 2005 John Wiley & Sons, Ltd.

  13. 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

  14. Temporary Sutureless Amniotic Membrane Patch for Acute Alkaline Burns

    Science.gov (United States)

    Kheirkhah, Ahmad; Johnson, Daniel A.; Paranjpe, Deval R.; Raju, V.K.; Casas, Victoria; Tseng, Scheffer C. G.

    2010-01-01

    Objective To evaluate the clinical outcome of a new sutureless approach for a temporary amniotic membrane patch (ProKera; Bio-Tissue, Inc, Miami, Florida) in eyes with acute burns. Methods Retrospective review of 5 eyes of 5 patients with grades I to III acute alkaline burns, receiving ProKera insertion within 8 days of injury. Results These eyes had either total (2 cases) or extensive (60%–75%, 3 cases) corneal epithelial defects with limbal (120°–360°) and conjunctival (30%–60%) epithelial defects. ProKera was inserted within a mean (SD) of 3.7 (3.1) days after burn and repeated 1 to 3 times for 3 cases. Conjunctival defects reepithelialized in 8.2 (5) days (range, 5–17 days), while limbal and corneal defects healed in 13.6 (8.3) days (range, 5–25 days). The latter was completed with circumferential closure of limbal defects followed by centripetal healing of corneal defects. In 3 eyes, early peripheral corneal neovascularization was followed by marked regression on completion of healing. During 16.8 (10.8) months of follow-up, all eyes retained a stable surface with improved corneal clarity, and without limbal deficiency or symblepharon. Conclusion This sutureless application of an amniotic membrane patch allows for early delivery of its biologic actions, which may help preserve remaining limbal stem cells for rapid expansion and prevent late cicatricial complications in eyes with mild and moderate acute alkaline burns. PMID:18695099

  15. 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....

  16. Amniotic membrane transplantation for acute ocular chemical burns in a child.

    Science.gov (United States)

    Thanikachalam, S; Kaliaperumal, Subashini; Srinivasan, Renuka; Sahu, Pramod Kumar

    2011-08-01

    An ocular burn injury with calcium hydroxide with opaque cornea and limbal ischaemia of more than 270 degrees which was treated byamniotic membrane transplantation on the 6th day following injury is reported. Postoperatively the ocular surface remained stable with no inflammation, vascularisation or infection. Amniotic membrane restored conjunctival surface much earlier than corneal surface and prevented symblepharon formation. We believe that amniotic membrane transplantation may be considered in acute phase of severe chemical injury for a more favourable prognosis.

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

    OpenAIRE

    Shadi Rezai; Justin Faye; Annika Chadee; Sri Gottimukkala; Ruchi Upadhyay; Carla Lara; Rajegowda, Benamanahalli H.; Corwin, Andrew D.; Rasila V. Lala; Jessica Vernon; Dilfuza Nuritdinova; Stephen Chasen; Cassandra E. Henderson

    2016-01-01

    Introduction. Amniotic band syndrome and sequence are a relatively rare condition in which congenital anomalies occur as a result of the adherence and entrapment of fetal parts with coarse fibrous bands of the amniotic membrane. A large percentage of reported cases have an atypical gestational history. The frequency of this obstetric complication is not affected by fetal gender, genetic abnormality, or prenatal infection. Case. A 21-year-old, G1P0 female parturient at 18 weeks and 5 days with...

  18. Bilateral fibular hemimelia with recto-vestibular fistula associated with amniotic bands.

    Science.gov (United States)

    Datta, K; Panja, S; Sarkar, Sumantra; Ghosh, Prakash C; Mirza, A

    2008-07-01

    Fibular Hemimelia is a rare congenital disorder of deficiency to complete absence of fibula. It has been associated with post axial skeletal anomalies like shortening of femur, absence of lateral rays of foot and syndactyle, polydactyle etc. We report this case because of associated amniotic constriction bands over both the forearm, suggesting amniotic bands may be one of the causal factors of Fibular Hemimelia due to insult to the growing limb bud.

  19. 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.

  20. Basement membrane dissolution and reassembly by limbal corneal epithelial cells expanded on amniotic membrane.

    Science.gov (United States)

    Li, Wei; He, Hua; Kuo, Ching-Liang; Gao, Yingying; Kawakita, Tetsuya; Tseng, Scheffer C G

    2006-06-01

    To investigate basement membrane (BM) formation during ex vivo expansion of limbal corneal epithelial cells on intact amniotic membrane (iAM) and epithelially denuded (d)AM. Human limbal explants were cultured on iAM and dAM. Expression of BM components, including laminin-5, type IV collagen, type VII collagen, perlecan, integrin alpha6, and epithelial cell differentiation markers such as p63, cytokeratin 3 (K3), and cytokeratin 12 (K12), were investigated by immunostaining. Levels of matrix metalloproteinase (MMP)-2 and MMP-9 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 in the conditioned media were determined by ELISA and gelatin zymography. All four BM components were preserved in both iAM and dAM before culturing, but dissolved 1 week afterward when MMP-2 was increased. Epithelial outgrowth correlated with increased expression of MMP-2 and -9 for both cultures. Resynthesis of BM began with laminin-5 followed by other components. This process took place at 1 week on iAM but at 2 weeks on dAM after culturing. At 4 weeks, BM was more maturely deposited as a linear band from the explant toward the leading edge on iAM and temporally correlated with a sharp decline of MMP-9 levels. In contrast, such BM deposition began at the leading edge on dAM only when TIMP-1 levels were increased. Epithelial cell outgrowth on iAM expressed more p63 but less K3 and K12 than did that on dAM. After dissolution of original amniotic BM, new BM formed by ex vivo expanded human limbal corneal epithelial cells on iAM deposits much faster and is more mature, resulting in regeneration of a limbal epithelial phenotype. In contrast, BM deposition is delayed and remains immature on dAM, resembling wound healing by a corneal epithelial phenotype. Thus, BM resynthesis may be used as another objective readout for assessing the success of ex vivo expansion of limbal epithelial progenitor cells on AM.

  1. New clinical application of amniotic membrane transplant for ocular surface disease.

    Science.gov (United States)

    Gheorghe, A; Pop, M; Burcea, M; Serban, M

    2016-01-01

    The new defined anatomical and functional complex conjunctiva-limbus-cornea is a new concept, which helps clinicians better understand and treat ocular surface pathologies. The management of the ocular surface disease has changed dramatically over the years, with spectacular improvements of techniques, and of course, results. The amniotic membrane, used as a graft or as a substrate for the cultivation of limbal corneal cells has showed encouraging results. To investigate the usefulness of amniotic membrane transplantation in ocular surface pathologies. The study is retrospective. 28 eyes of 28 patients with ocular surface pathologies (ocular burns, recent and old, symblepharon, pterygium, corneal and conjunctival tumors, persistent epithelial defect) underwent an amniotic membrane transplantation during a 1 year period. The necrotic and the scar tissue were first excised in all the patients and the amniotic membrane was sutured with an epithelial face up. Follow up ranged from 1 to 12 months. Good results were obtained in all 28 eyes. The anatomy of cornea and conjunctiva was improved, with limited benefits only in old ocular burn, symblepharon and in one case of extended tumors of the cornea and conjunctiva. Out of 28 eyes, 23 (82%) also had a visual acuity improvement. The amniotic membrane may be considered a good alternative for ocular surface reconstruction especially in acute status. AMT = amniotic membrane transplantation, BCVA = best corrected visual acuity, BUT = break up time.

  2. 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.

  3. [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.

  4. A new surgical technique to treat corneal perforations using amniotic membrane and surgical adhesive

    Directory of Open Access Journals (Sweden)

    K.C.S. Pontes

    2014-06-01

    Full Text Available The objectives of this study were to compare surgical techniques and the effects of using n-butyl 2-cyanoacrylate and bovine amniotic membrane to repair perforated lesions in corneas. Penetrating keratoplasty was performed in sixty New Zealand White rabbits under general anesthesia. Group 1 (G1 was treated with n-butyl 2-cyanoacrylate, group 2 (G2 received a fragment of amniotic membrane through the anterior chamber and application of n-butyl 2-cyanoacrylate over the lesion, group 3 (G3 was treated with the same technique as G2 with the addition of an amniotic membrane bandage covering the cornea and sutured in the limbus region, and group 4 (G4 was treated with an amniotic membrane sutured to the lesion and an amniotic membrane bandage sutured in the limbus region. Clinical, histological and histomorphometric examinations of the corneas were performed. The membrane acted as a barrier for aqueous humor in G2 and G3, thereby keeping the surface dry for adhesive application; it also prevented the adhesive from contacting intraocular structures. The groups treated with amniotic membrane and surgical adhesive showed better results than the groups treated with either material alone. Thus, the combination of the membrane with the adhesive is recommended for this type of lesion.

  5. The Ratio of Testosteron and Follicle Stimulating Hormone in the Amniotic Fluid

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Suk Shin [Hanyang University College of Medicine, Seoul (Korea, Republic of)

    1982-09-15

    To evaluate fetal sex-hormonal status before delivery, testosterone and follicle stimulating hormone (FSH) levels were measured in 64 amniotic fluid samples at midgestation by radioimmunoassay method. The mean concentration of testosterone in amniotic fluid of 37 cases carrying male fetus was 90.7 pg/ml and 27 cases carrying female fetus was 62.3 pg/ml. The mean amniotic fluid FSH concentration of male fetus was 1.15 mIU/ml and of female fetus was 11.98 mIU/ml. The amniotic fluid testosterone and FSH' concentrations had statistical difference between male and female fetuses. The ratio of testosterone over FSH in the amniotic fluid was 231.2 in female, 9.8 in female respectively and very significant difference was noticed. The levels of testosterone/FSH greater than 25 were found over 92% of male fetus and lesser than 25 were found over 92% female fetus. Measurement of testosterone and FSH especially testosterone/FSH ratio in amniotic fluid in midgestation may be an adjunct to other method of fetal sex determination.

  6. The diagnostic performance of the Mass Restricted (MR) score in the identification of microbial invasion of the amniotic cavity or intra-amniotic inflammation is not superior to amniotic fluid interleukin-6.

    Science.gov (United States)

    Romero, Roberto; Kadar, Nicholas; Miranda, Jezid; Korzeniewski, Steven J; Schwartz, Alyse G; Chaemsaithong, Piya; Rogers, Wade; Soto, Eleazar; Gotsch, Francesca; Yeo, Lami; Hassan, Sonia S; Chaiworapongsa, Tinnakorn

    2014-05-01

    Intra-amniotic infection/inflammation are major causes of spontaneous preterm labor and delivery. However, diagnosis of intra-amniotic infection is challenging because most are subclinical and amniotic fluid (AF) cultures take several days before results are available. Several tests have been proposed for the rapid diagnosis of microbial invasion of the amniotic cavity (MIAC) or intra-amniotic inflammation. The aim of this study was to examine the diagnostic performance of the AF Mass Restricted (MR) score in comparison with interleukin-6 (IL-6) and matrix metalloproteinase-8 (MMP-8) for the identification of MIAC or inflammation. AF samples were collected from patients with singleton gestations and symptoms of preterm labor (n = 100). Intra-amniotic inflammation was defined as >100 white blood cells/mm(3) (WBCs) in AF; MIAC was defined as a positive AF culture. AF IL-6 and MMP-8 were determined using ELISA. The MR score was obtained using the Surface-Enhanced Laser Desorption Ionization Time of Flight (SELDI-TOF) mass spectrometry. Sensitivity and specificity were calculated and logistic regression models were fit to construct receiver-operating characteristic (ROC) curves for the identification of each outcome. The McNemar's test and paired sample non-parametric statistical techniques were used to test for differences in diagnostic performance metrics. (1) The prevalence of MIAC and intra-amniotic inflammation was 34% (34/100) and 40% (40/100), respectively; (2) there were no significant differences in sensitivity of the three tests under study (MR score, IL-6 or MMP-8) in the identification of either MIAC or intra-amniotic inflammation (using the following cutoffs: MR score >2, IL-6 >11.4 ng/mL, and MMP-8 >23 ng/mL); (3) there was no significant difference in the sensitivity among the three tests for the same outcomes when the false positive rate was fixed at 15%; (4) the specificity for IL-6 was not significantly different from that of the MR score in

  7. Amniotic membrane extract ameliorates benzalkonium chloride-induced dry eye in a murine model.

    Science.gov (United States)

    Xiao, Xinye; Luo, Pingping; Zhao, Hui; Chen, Jingyao; He, Hui; Xu, Yuxue; Lin, Zhirong; Zhou, Yueping; Xu, Jianjiang; Liu, Zuguo

    2013-10-01

    Human amniotic membrane (AM) is avascular but contains various beneficial bioactive factors, its extract (AE) is also effective in treating many ocular surface disorders. In this study, we for the first time evaluated the therapeutic effects of AE on dry eye induced by benzalkonium chloride in a BALB/c mouse model. Topical application of AE (1.5 and 3 μg/eye/day) resulted in significantly longer tear break-up time on Day 3 and 6, lower fluorescein staining scores on Day 3, and lower inflammatory index on Day 6. AE reduced corneal epithelial K10 expression, inflammatory infiltration, and levels of TNF-α, IL-1β and IL-6 in BAC treated mice than that in the control mice. Moreover, decreased TUNEL positive cells in cornea and increased goblet cells in conjunctiva were also observed in AE treated corneas. Finally, AE induced more Ki-67 positive cells in corneal epithelium of dry eye mouse. Taken together, our data provide further support for BAC induced dry eye model as a valuable for dry eye study and suggest a great potential for AE as a therapeutic agent in the clinical treatment of dry eye. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. "Pleats fold" technique of amniotic membrane transplantation for management of corneal perforations.

    Science.gov (United States)

    Namba, Hiroyuki; Narumi, Mari; Nishi, Katsuhiro; Goto, Sakiko; Hayashi, Shion; Yamashita, Hidetoshi

    2014-06-01

    The aim of this study was to examine the efficacy and surgical success rates of amniotic membrane (AM) transplantation performed for corneal perforation closure using a novel technique. This study included 6 eyes from 6 patients with corneal perforation who had received AM transplantation between May 2011 and April 2012. The AM was collected from human placenta shortly after cesarean section. In surgery, the AM was folded into pleats and used to plug the wound using 10-0 nylon suture. The wound was then covered with an AM seal. After reepithelialization and AM scarring, sutures were removed. All 6 patients had successful wound closure with 1 surgery. One patient underwent optical keratoplasty later, and 1 patient required combined preserved sclera transplantation. The absolute value of astigmatism decreased to incised wounds. Our "Pleats Fold" AM transplantation technique can achieve definite closure and effectively repair wounds of various sizes. Postoperative astigmatic values were acceptable. Therefore, we recommend this procedure for repairing lesions <3 mm in diameter that do not involve the central cornea and that are infection free.

  9. 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.

  10. Satellited 4q identified in amniotic fluid cells

    Energy Technology Data Exchange (ETDEWEB)

    Miller, I.; Hsieh, C.L.; Songster, G. [Stanford Univ. Medical Center, Stanford, CA (United States)] [and others

    1995-01-16

    Extra material was identified on the distal long arm of a chromosome 4 in an amniotic fluid specimen sampled at 16.6 weeks of gestational age. There was no visible loss of material from chromosome 4, and no evidence for a balanced rearrangement. The primary counseling issue in this case was advanced maternal age. Ultrasound findings were normal, and family history was unremarkable. The identical 4qs chromosome was observed in cells from a paternal peripheral blood specimen and appeared to be an unbalanced rearrangement. This extra material was NOR positive in lymphocytes from the father, but was negative in the fetal amniocytes. Father`s relatives were studied to verify the familial origin of this anomaly. In situ hybridization with both exon and intron sequences of ribosomal DNA demonstrated that ribosomal DNA is present at the terminus of the 4qs chromosome in the fetus, father, and paternal grandmother. This satellited 4q might have been derived from a translocation event that resulted in very little or no loss from the 4q and no specific phenotype. This derivative chromosome 4 has been inherited through at least 3 generations of phenotypically normal individuals. 8 refs., 3 figs.

  11. Amniotic membrane extraction solution for ocular chemical burns.

    Science.gov (United States)

    Liang, Lingyi; Li, Wei; Ling, Shiqi; Sheha, Hosam; Qiu, Weiqiang; Li, Chaoyang; Liu, Zuguo

    2009-12-01

    To evaluate the efficacy of amniotic membrane extract (AME) for ocular chemical burns. Prospective non-comparative interventional case series study. Consecutive 14 eyes of 11 patients with acute or chronic chemical burns, being recruited in one referral centre, received AME topically in combination with traditional treatment. Ocular discomfort, visual acuity, ocular surface inflammation, re-epithelialization, corneal thickness, corneal neovascularization and symblepharon were evaluated. Symptom relieved and ocular surface inflammation reduced dramatically in all eyes. Epithelial defect healed in all eyes with acute burns, in which less than 7 clock hours of limbus was involved, after 16.6 days (1-44 days) AME treatment. AME failed to close the epithelial defect in all eyes with chronic chemical burn coexisting diffuse limbal stem cell deficiency; however, the area of epithelial defect decreased to 58% (11.1%-68.2%) at final visit. During a follow-up period of 8.2 months (6-11 months), visual acuity improved in 12 eyes (86%). There was mild neovascularization in three eyes with grade III and IV acute burns, and slow progress of neovascularization in chronic burns. Mild symblepharon developed in two eyes with grade III and IV acute burns, whereas there was no significant progress of symblepharon in chronic cases. Although it is a preliminary and uncontrolled study, topical application of AME is effective in reducing inflammation, promoting reepithelization in the treatment of chemical burns, especially for mild to moderate acute cases.

  12. Reproductive decisions after the diagnosis of amniotic fluid embolism.

    Science.gov (United States)

    Moaddab, Amirhossein; Klassen, Miranda; Priester, Cynthia D; Munoz, Elvira H; Belfort, Michael A; Clark, Steven L; Dildy, Gary A

    2017-04-01

    This study aims to describe the subsequent reproductive outcomes in women who either correctly or incorrectly were diagnosed with amniotic fluid embolism (AFE). Medical records were obtained, abstracted and reviewed by authors with extensive experience in critical care obstetrics. Telephone interviews of all survivors were conducted to determine obstetrical and contraceptive history. A subgroup underwent further telephone interview to address subsequent reproductive decisions. By November 2015, 116 medical records of patients diagnosed with AFE were reviewed. Patients who had undergone hysterectomy (n=26), died (n=9), or developed Sheehan's syndrome (n=1) at the time of the original event were excluded from the present analysis. Of the remaining 80 women, 30% (24/80) had subsequently conceived and 32.5% (26/80) patients or their partners had undergone permanent sterilization. At the time of this report, 66% (21/32) of registry participants were categorized to have had AFE and 34% (11/32) as not likely AFE or indeterminate. The syndrome of AFE is over-diagnosed. Women diagnosed with AFE who survive conceive another pregnancy less frequently than US women over similar time intervals and often choose a permanent sterilization method, whether or not they actually had AFE, largely out of fear of AFE recurrence. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Incidence, risk factors, and consequences of amniotic fluid embolism.

    Science.gov (United States)

    Kramer, Michael S; Abenhaim, Haim; Dahhou, Mourad; Rouleau, Jocelyn; Berg, Cynthia

    2013-09-01

    Amniotic fluid embolism (AFE) is a rare but serious cause of maternal mortality whose aetiology remains obscure. Previous population-based studies have reported associations with labour induction and caesarean delivery. We updated a previous analysis based on the US Nationwide Inpatient Sample from 1999 to 2008. We adapted a diagnostic validation algorithm to minimise false-positive diagnoses, along with statistical methods that account for the stratified random sampling design. Of the 8 571 209 deliveries recorded in the database, 276 met our case definition of AFE, of which 62 (22.9% of the 274 with known vital status) were fatal. Significant associations with AFE were observed for medical induction {adjusted odds ratio [aOR] = 1.7 [95% confidence interval (CI) 1.2, 2.5]}, caesarean delivery [aOR = 15.0; 95% CI 9.4, 23.9], instrumental vaginal delivery [aOR = 6.6; 95% CI 4.0, 11.1], and cervical/uterine trauma [aOR = 7.4; 95% CI 3.6, 14.9]. AFE was associated with increases in risk of stillbirth, hysterectomy, maternal death, and prolonged maternal length of delivery hospital stay. AFE remains an extremely serious obstetric complication with high risks of maternal and fetal mortality. The increased risks of AFE associated with labour induction and caesarean delivery have implications for elective use of these interventions. © 2013 John Wiley & Sons Ltd.

  14. Surgical amniotic membrane transplantation after conjunctival and limbal tumor excision

    Directory of Open Access Journals (Sweden)

    Seniz Engur Goktas

    Full Text Available ABSTRACT Purpose: To evaluate the clinical results of patients treated by amniotic membrane transplantation (AMT following excision of conjunctival and limbal tumors. Methods: A total of 14 eyes of 14 patients who underwent AMT after total lesion-free tumor excision and perilesional cryotherapy were evaluated. Results: The excised tumors comprised 7 conjunctival intraepithelial neoplasia, 5 conjunctival nevus, 1 primary acquired melanosis, and 1 squamous cell carcinoma. Limbus was involved in 10 cases, whereas cornea was involved in 6 cases. The average measurement of the tumor base was 14.8 mm (range 6-20 mm, SD 16 mm. The mean follow-up time period was 17.5 months (range 6 -60 months, SD 20 months. Complete healing occurred in eight eyes, but limbal cell deficiency developed in two eyes. Four cases had recurrence and were treated with the same surgical and medical procedures; during follow-up after recurrence, superficial peripheral vascularization and corneal scar were noted to have developed. Conclusions: AMT was effective for reconstruction of tissue defect after excision and cryotherapy of limbal and conjunctival tumors. In most of these cases, complete healing was achieved with a smooth, stable, and translucent surface.

  15. 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

    inhibitory peptide and glucagon-like peptide 2, changed gut microbiota, and reduced intestinal permeability. There were no effects on GI weight, percentage mucosa, villus height, plasma citrulline, hexose absorptive capacity, and digestive enzymes. Intestinal interleukin (IL)-1β levels and expression of IL1B......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...

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

    Science.gov (United States)

    Rezai, Shadi; Faye, Justin; Chadee, Annika; Gottimukkala, Sri; Upadhyay, Ruchi; Lara, Carla; Rajegowda, Benamanahalli H; Corwin, Andrew D; Lala, Rasila V; Vernon, Jessica; Nuritdinova, Dilfuza; Chasen, Stephen; Henderson, Cassandra E

    2016-01-01

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

  17. Both Freshly Prepared and Frozen-Stored Amniotic Membrane Cells Express the Complement Inhibitor CD59

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    Ágnes Füst

    2012-01-01

    Full Text Available Amniotic membrane proved to be very effective tool in the treatment of a number of ocular surface diseases. The amniotic membrane, however, has to be stored before its transplantation onto the ocular surface followed by mandatory serologic control in order to exclude the transmission of certain viruses. Therefore it is most important to study if cryopreservation of the membrane affects cell surface expression of the molecules. We measured cell surface expression of CD59, a membrane-bound complement inhibitor on the cells of freshly prepared and cryopreserved amniotic membrane. Cells of amniotic membrane were separated mechanically. Epithelial and mesenchymal cells were identified by the intracellular expression of nanog and the cell surface ICAM1 positivity, respectively. Multicolor flow cytometric immunophenotyping was used for determination of the CD59 expression. CellQuest-Pro software program (Becton Dickinson was used both for measurements and analysis. CD59-positive cells could be detected in all investigated samples and in all investigated cell types, although the expression level of CD59 differed. CD59 was expressed both on freshly prepared and frozen-stored samples. Higher level of CD59 was detected on ICAM1+ mesenchymal cells than on nanog+ epithelial cells. Our findings indicate that amniotic membranes maintain their complement inhibiting capacity after cryopreservation.

  18. [Measuring the turbidity of amniotic fluid, a possibility to assess fetal maturity before birth (author's transl)].

    Science.gov (United States)

    Lamberti, G

    1978-08-01

    At the end of gestation, depending on maturation of the fetus and especially of its skin, vernix caseosa is detached into amniotic fluid. The changes of amniotic fluid turbidity can be quantitatively verified with a fotometer (filter for 578 nm, 10 mm disposable cuvettes). The turbidity of amniotic fluid was measured in 125 cases. No correlation was found between turbidity and length of gestation from day 261 to day 287 after last menstruation. In contrast, there is strong and significant contingency between turbidity of amniotic fluid and maturity of fetal skin (quantity of vernix caseosa) or clinical maturity assessed according to Farr et al. Finding a turbidity below 0.5 U on extinction scale of the fotometer, the fetus is not fully mature in 50% of these cases; placental dysfunction can be excluded with high probability. In contrast, two thirds of the children with high turbidity of amniotic fluid (more than 1.0 U on extinction scale) had clinical signs of placental dysfunction.

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

    Directory of Open Access Journals (Sweden)

    Shadi Rezai

    2016-01-01

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

  20. Review: Impact of mediators present in amniotic fluid on preterm labour.

    Science.gov (United States)

    Vrachnis, Nikolaos; Karavolos, Stamatios; Iliodromiti, Zoe; Sifakis, Stavros; Siristatidis, Charalambos; Mastorakos, George; Creatsas, George

    2012-01-01

    Preterm birth continues to be one of the most important issues in current obstetric medicine, being the single largest cause of perinatal morbidity and mortality. The signals that initiate preterm and term labour remain a mystery. Intrauterine inflammation with the secretion of cytokines is one of the accepted explanations for the mechanism of initiation of preterm labour. This review discusses the current understanding of the molecular mechanisms for the initiation of preterm labour, focusing chiefly on the role of intra-amniotic fluid mediators, whether endogenous or infection-induced, in the regulation of inflammatory response pathways associated with spontaneous preterm labour. Prostaglandins (PGs) are considered to be one of the key mediators of preterm labour, with the concentration of biologically active PGs in the amniotic fluid, particularly PGE(2) and PGF(2α), being significantly higher in women with preterm labour. Cytokines, such as interleukins and tumour necrosis factor alpha, additionally play a dominant role in preterm labour, particularly in association with infection. Elevated amniotic fluid concentrations of extracellular matrix mediators, including metalloproteases, are also implicated in the process of foetal membrane rupture in preterm labour. Allelic variations in the main amniotic fluid mediators may be the key to understanding the disparity in the rates of preterm birth between different ethnic populations. We also discuss the role of other potential mediators such as cell-adhesion molecules, nitric oxide and novel biomarkers found in the amniotic fluid.

  1. 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.

  2. Single and multilayer amniotic membrane transplantation for persistent corneal epithelial defect with and without stromal thinning and perforation.

    Science.gov (United States)

    Prabhasawat, P; Tesavibul, N; Komolsuradej, W

    2001-12-01

    To evaluate the efficacy of amniotic membrane transplantation (AMT) in persistent corneal epithelial defect with or without stromal thinning and corneal perforation. 28 patients (28 eyes) with persistent corneal epithelial defect unresponsive to medical treatment were given preserved human amniotic membrane transplants. The patients were divided into three groups: group A, persistent corneal epithelial defect 10 eyes; group B, epithelial defect with stromal thinning 13 eyes; and group C, corneal perforation five eyes. AMT was performed using one layer in group A and multilayers in group B and C. The causes of persistent epithelial defect were neurotrophic keratopathy (24 eyes), limbal deficiency (six eyes), exposure keratopathy (four eyes), and Mooren's ulcer (one eye). Success was noted in 82.1% (23/28 eyes) in all groups, with 80% (8/10 eyes), 84.6% (11/13 eyes), and 80% (4/5 eyes) in groups A, B, and C respectively, with a mean follow up of 10.9 months (1-30 months). The mean epithelialisation time after AMT was 2.1 weeks. The healing times of groups B and C are also significantly shorter than group A (p=0.017 and 0.018, respectively). Corneal stromal thickness was significantly increased in all cases in groups B and C (p=0.006). Those with corneal perforation in group C were completely healed by multilayer AMT. There was no difference in the epithelialisation time between successful cases treated by a single operation (17 eyes) or repeated operation (six eyes). Vision improved in 18.9% (8/28 eyes) and worsened as a result of cataract formation in 2.3% (1/28 eyes). Failure was noted in 17.9% (5/28 eyes), because of corneal infection (two eyes), neurotrophic keratopathy with and without limbal deficiency (two eyes), and intractable corneal perforation (one eye). No patient developed major immediate postoperative complications or graft rejection. Amniotic membrane can successfully treat refractory corneal epithelial defect by promoting epithelial healing and thus

  3. 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.

  4. In utero therapy for congenital disorders using amniotic fluid stem cells

    Science.gov (United States)

    Ramachandra, Durrgah L.; Shaw, Steven S. W.; Shangaris, Panicos; Loukogeorgakis, Stavros; Guillot, Pascale V.; Coppi, Paolo De; David, Anna L.

    2014-01-01

    Congenital diseases are responsible for over a third of all pediatric hospital admissions. Advances in prenatal screening and molecular diagnosis have allowed the detection of many life-threatening genetic diseases early in gestation. In utero transplantation (IUT) with stem cells could cure affected fetuses but so far in humans, successful IUT using allogeneic hematopoietic stem cells (HSCs), has been limited to fetuses with severe immunologic defects and more recently IUT with allogeneic mesenchymal stem cell transplantation, has improved phenotype in osteogenesis imperfecta. The options of preemptive treatment of congenital diseases in utero by stem cell or gene therapy changes the perspective of congenital diseases since it may avoid the need for postnatal treatment and reduce future costs. Amniotic fluid stem (AFS) cells have been isolated and characterized in human, mice, rodents, rabbit, and sheep and are a potential source of cells for therapeutic applications in disorders for treatment prenatally or postnatally. Gene transfer to the cells with long-term transgenic protein expression is feasible. Recently, pre-clinical autologous transplantation of transduced cells has been achieved in fetal sheep using minimally invasive ultrasound guided injection techniques. Clinically relevant levels of transgenic protein were expressed in the blood of transplanted lambs for at least 6 months. The cells have also demonstrated the potential of repair in a range of pre-clinical disease models such as neurological disorders, tracheal repair, bladder injury, and diaphragmatic hernia repair in neonates or adults. These results have been encouraging, and bring personalized tissue engineering for prenatal treatment of genetic disorders closer to the clinic. PMID:25566071

  5. Amniotic membrane transplantation as an adjunct to medical therapy in acute ocular burns.

    Science.gov (United States)

    Tandon, Radhika; Gupta, Noopur; Kalaivani, Mani; Sharma, Namrata; Titiyal, Jeewan S; Vajpayee, Rasik B

    2011-02-01

    To evaluate the role of amniotic membrane transplantation in patients with acute ocular burns. In a prospective, randomised, controlled clinical trial, 100 patients with grade II to IV acute ocular burns (Roper Hall Classification) were recruited. 50 patients with grade II-III burns were graded as moderate burns, and 50 patients with grade IV burns were graded as severe burns. Both groups were individually randomised into control group (n=25) and study group (n=25). The corresponding grade of ocular surface burn by Dua classification was noted. The eyes in the study group underwent amniotic membrane transplantation in addition to conventional medical therapy. In the control group, conventional medical therapy along with mechanical release of early adhesions as and when necessary was instituted. Rate of healing of corneal epithelial defect, visual acuity, extent of corneal vascularisation, corneal clarity and formation of symblepharon were compared in both groups. In patients with moderate ocular burns treated with amniotic membrane transplantation, the rate of epithelial healing was significantly better than the group treated with standard medical therapy alone (p=0.0004). There was no overall difference in the final visual outcome, symblepharon formation, corneal clarity and vascularisation with or without amniotic membrane transplantation. Amniotic membrane transplantation in eyes with acute ocular burns promotes faster healing of epithelial defect in patients with moderate grade burns. There seems to be no definite long-term advantage of amniotic membrane transplantation over medical therapy and mechanical release of adhesions in terms of final visual outcome, appearance of symblepharon and corneal vascularisation when compared in a controlled clinical setting.

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

    Directory of Open Access Journals (Sweden)

    Lin Li

    2015-01-01

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

  7. [Amniotic membrane transplantation in proven ulcerative herpetic keratitis: successful anti-inflammatory treatment in time].

    Science.gov (United States)

    Spelsberg, H; Reichelt, J A

    2008-01-01

    In active herpetic keratitis, treatment with systemic and topical aciclovir together with low dosed-out steroids has been successful for few decades. However, in cases with severe herpetic ulcer and melting inflammation, emergency penetrating keratoplasty is necessary, which carries high risk for recurrent herpetic keratitis and rejection. The anti-inflammatory effect of amniotic membrane transplantation can be used as a first surgical step in order to gain time for planned penetrating keratoplasty. The results are analysed in this retrospective case series. Twelve eyes of 12 patients with clinically highly active herpetic ulcer without healing despite topical and systemic aciclovir were treated with amniotic membrane transplantation. Ten of 12 eyes (83 %) showed positive PCR testing for HSV-1. In one case, the molecular genetic result was negative, one analysis for HSV was not done. For surgery, debridement of the margins and the ground of the ulcer was done with steam heated cauterization. Afterwards, an amniotic membrane was made to cover the defect and fixed with single sutures. Peri- and postoperatively, the patients were treated with systemic aciclovir (4000 mg daily one week, 2000 mg daily 2 weeks) and gentamicin ointment topically. Mean follow-up was 6.8 months (1-12.5). After 25 days in the mean (11-34), nine of 12 patients experienced an intact epithelial corneal surface together with a clinically clear reduction of the inflammation. Three other patients did not recover and were treated with emergency penetrating keratoplasty after 31 days in mean (25-35). Visual acuity did not improve after amniotic membrane transplantation except in one case. The known anti-inflammatory potential of amniotic membrane causes a reduction of the inflammatory process in herpetic ulcer. A precondition is a sufficient antiherpetic and anti-inflammatory therapy and a decision for amniotic membrane transplantation in time in order to avoid an emergency penetrating keratoplasty

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

    Directory of Open Access Journals (Sweden)

    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

  9. The evolution of body size, Cope's rule and the origin of amniotes.

    Science.gov (United States)

    Laurin, Michel

    2004-08-01

    The evolution of body size in tetrapods is assessed using a database that includes 107 early stegocephalian species ranging in time from the Frasnian (Upper Devonian) to the Tatarian (Upper Permian). All analyses use methods that incorporate phylogenetic information (topology and branch lengths). In all tests, the impact of alternative topologies and branch lengths are assessed. Previous reports that raised doubts about the accuracy of squared-change parsimony assessment of ancestral character value appear to have used datasets in which there was no phylogenetic signal. Hence, squared-change parsimony may be more reliable than suggested in recent studies, at least when a phylogenetic signal is present in the datasets of interest. Analysis using random taxon reshuffling on three reference phylogenies shows that cranial and presacral length include a strong phylogenetic signal. Character optimization of body size in stegocephalians using squared-change parsimony on a time-calibrated phylogeny incorporating branch length information is used to test a previously published scenario on the origin of amniotes and of the amniotic egg that implies that the ancestors of amniotes were small (no more than 10 cm in snout-vent length), and that their size increased subsequent to the appearance of the amniotic egg. The optimization suggests that first amniotes were somewhat larger than previously hypothesized; the estimated snout-vent length is about 24 cm, and the lower end of the 95% confidence interval of the phylogeny that yields the smallest inferred size suggests that no ancestor of amniotes measured less than 12 cm in snout-vent length. Character optimization, permutational multiple linear regressions, and independent contrast analyses show that Cope's rule of phyletic size increase applies to early reptiliomorphs but that it does not apply to early stegocephalians globally.

  10. 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.

  11. Clinical chorioamnionitis at term III: how well do clinical criteria perform in the identification of proven intra-amniotic infection?

    Science.gov (United States)

    Romero, Roberto; Chaemsaithong, Piya; Korzeniewski, Steven J; Kusanovic, Juan P; Docheva, Nikolina; Martinez-Varea, Alicia; Ahmed, Ahmed I; Yoon, Bo Hyun; Hassan, Sonia S; Chaiworapongsa, Tinnakorn; Yeo, Lami

    2016-01-01

    The diagnosis of clinical chorioamnionitis is based on a combination of signs [fever, maternal or fetal tachycardia, foul-smelling amniotic fluid (AF), uterine tenderness and maternal leukocytosis]. Bacterial infections within the amniotic cavity are considered the most frequent cause of clinical chorioamnionitis and an indication for antibiotic administration to reduce maternal and neonatal morbidity. Recent studies show that only 54% of patients with the diagnosis of clinical chorioamnionitis at term have bacteria in the AF and evidence of intra-amniotic inflammation. The objective of this study was to examine the performance of the clinical criteria for the diagnosis of chorioamnionitis to identify patients with microbial-associated intra-amniotic inflammation (also termed intra-amniotic infection). This retrospective cross-sectional study included 45 patients with the diagnosis of clinical chorioamnionitis at term, whose AF underwent analysis for: 1) the presence of microorganisms using both cultivation and molecular biologic techniques [polymerase chain reaction (PCR) with broad primers], and 2) interleukin (IL)-6 concentrations by enzyme-linked immunosorbent assay. The diagnostic performance (sensitivity, specificity, accuracy, and likelihood ratios) of each clinical sign and their combination to identify clinical chorioamnionitis were determined using microbial-associated intra-amniotic inflammation [presence of microorganisms in the AF using cultivation or molecular techniques and elevated AF IL-6 concentrations (≥2.6 ng/mL)] as the gold standard. The accuracy of each clinical sign for the identification of microbial-associated intra-amniotic inflammation (intra-amniotic infection) ranged between 46.7% and 57.8%. The combination of fever with three or more clinical criteria did not substantially improve diagnostic accuracy. In the presence of a fever during labor at term, signs used to diagnose clinical chorioamnionitis do not accurately identify the

  12. [The clinical analysis of 38 cases with amniotic fluid embolism].

    Science.gov (United States)

    Yang, W; Zhou, N; Zhou, Y

    2000-02-01

    To analyze the clinical course of amniotic fluid embolism (AFE) and identify the high risk factors. Thirty-eight cases diagnosed as AFE in Suzhou region during period of past 15 years were analyzed retrospectively. Fifteen years were divided into 5 stages with 3 years each. Of 38 cases, 30 (78.9%) were primigravida and one twins. There were 34 maternal deaths. Among them, 31 died in the first four stages and 3 died in the last stage. There were 4 cases survived from AFE in the last stage. AFE accounted for 15% of total maternal deaths in the past 15 years, which is the second cause of maternal death. Of 38 cases, 2 (5.2%) occurred before 28 gestational weeks. 20 (52.6%) suffered from AFE during labour, 13 (34.2%) after delivery, and 3 (7.8%) before labour. All cases presented respiratory distress, cyanosis, chest discomfort and/or shock, cardiopulmonary collapse. Sixteen cases had postpartum hemorrhage and/or laboratory evidence of DIC. Of 38 cases, 15 (39.5%) died within one hour after onset of AFE. The predisposing factors for AFE included strong uterine contractions due to oxytocin or PEG augmented 17 cases (44.7%), pregnancy induced hypertension 11 cases (28.9%), multigravida and/or elder gravida 9 cases (23.7%) and cesarean section 8 cases (21.1%). AFE must be suspected in maternal cases with sudden collapse, especially the clinical symptoms of allergic reaction. Prevention of hypertonic uterine contraction and prompt resuscitation, and emergency surgical delivery can improve the prognosis of AFE.

  13. Amniotic membrane traps and induces apoptosis of inflammatory cells in ocular surface chemical burn

    Science.gov (United States)

    Liu, Ting; Zhai, Hualei; Xu, Yuanyuan; Dong, Yanling; Sun, Yajie; Zang, Xinjie

    2012-01-01

    Purpose Severe chemical burns can cause necrosis of ocular surface tissues following the infiltration of inflammatory cells. It has been shown that amniotic membrane transplantation (AMT) is an effective treatment for severe chemical burns, but the phenotypes of cells that infiltrate the amniotic membrane and the clinical significance of these cellular infiltrations have not previously been reported. The present work studies the inflammation cell traps and apoptosis inducing roles of the amniotic membrane after AMT in patients with acute chemical burns. Methods A total of 30 patients with acute alkaline burns were classified as having either moderate or severe burns. In all participants, AMT was performed within one week of his/her injury. After 7–9 days, the transplanted amniotic membranes were removed. Histopathological and immunohistochemical techniques were used for the examination and detection of infiltrating cells, and tests for the expression of CD (cluster of differentiation)15, CD68, CD3, CD20, CD57, CD31, CD147, and CD95 (Fas) were performed. A TUNEL (TdT-mediated dUTP nick end labeling) assay was used to confirm apoptosis of the infiltrating cells. Three patients with herpes simplex-induced keratitis who had undergone AMT to treat persistent epithelium defects were used as a control group. Amniotic membrane before transplantation was used as another control. Results After amniotic membrane transplantation, the number of infiltrating cells in patients with severe burns was significantly higher than in patients with moderate burns or in control patients (pburns patients, CD15 and CD68 were widely expressed in the infiltrating cells, and CD3, CD20, and CD57 were only found in a small number of cells. Occasionally, CD31-positive cells were found in the amniotic membranes. More cells that were CD147, Fas, and TUNEL positive were found in patients with severe burns than in patients with moderate burns or in control patients. Conclusions Neutrophils and

  14. 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

  15. Carbodiimide cross-linking of amniotic membranes in the presence of amino acid bridges.

    Science.gov (United States)

    Lai, Jui-Yang

    2015-06-01

    The purpose of this study was to investigate the carbodiimide cross-linking of amniotic membrane (AM) in the presence of amino acid bridges. The biological tissues were treated with glycine, lysine, or glutamic acid and chemically cross-linked to examine the role of amino acid types in collagenous biomaterial processing. Results of zeta potential measurements showed that the use of uncharged, positively and negatively charged amino acids dictates the charge state of membrane surface. Tensile strength and water content measurements demonstrated that the addition of lysine molecules to the cross-linking system can increase the cross-linking efficiency and dehydration degree while the introduction of glutamic acid in the AM samples decreases the number of cross-links per unit mass of chemically modified tissue collagen. The differences in the cross-linking density further determined the thermal and biological stability by differential scanning calorimetry and in vitro degradation tests. As demonstrated in matrix permeability studies, the improved formation of covalent cross-linkages imposed by lysine facilitated construction of stronger cross-linking structures. In contrast, the added glycine molecules were insufficient to enhance the resistances of the proteinaceous matrices to thermal denaturation and enzymatic degradation. The cytocompatibility of these biological tissue membranes was evaluated by using human corneal epithelial cell cultures. Results of cell viability, metabolic activity, and pro-inflammatory gene expression level showed that the AM materials cross-linked with carbodiimide in the presence of different types of amino acids are well tolerated without evidence of detrimental effect on cell growth. In addition, the amino acid treated and carbodiimide cross-linked AM implants had good biocompatibility in the anterior chamber of the rabbit eye model. Our findings suggest that amino acid type is a very important engineering parameter to mediate

  16. Augmented dried versus cryopreserved amniotic membrane as an ocular surface dressing.

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    Claire L Allen

    Full Text Available Dried amniotic membrane (AM can be a useful therapeutic adjunct in ophthalmic surgery and possesses logistical advantages over cryopreserved AM. Differences in preservation techniques can significantly influence the biochemical composition and physical properties of AM, potentially affecting clinical efficacy. This study was established to investigate the biochemical and structural effects of drying AM in the absence and presence of saccharide lyoprotectants and its biocompatibility compared to cryopreserved material.AM was cryopreserved or dried with and without pre-treatment with trehalose or raffinose and the antioxidant epigallocatechin (EGCG. Structural and visual comparisons were assessed using electron microscopy. Localisation, expression and release of AM biological factors were determined using immunoassays and immunofluorescence. The biocompatibility of the AM preparations co-cultured with corneal epithelial cell (CEC or keratocyte monolayers were assessed using cell proliferation, cytotoxicity, apoptosis and migration assays.Drying devitalised AM epithelium, but less than cryopreservation and cellular damage was reduced in dried AM pre-treated with trehalose or raffinose. Dried AM alone, and with trehalose or raffinose showed greater factor retention efficiencies and bioavailability compared to cryopreserved AM and demonstrated a more sustained biochemical factor time release in vitro. Cellular health assays showed that dried AM with trehalose or raffinose are compatible and superior substrates compared to cryopreserved AM for primary CEC expansion, with increased proliferation and reduced LDH and caspase-3 levels. This concept was supported by improved wound healing in an immortalised human CEC line (hiCEC co-cultured with dried and trehalose or raffinose membranes, compared to cryopreserved and fresh AM.Our modified preservation process and our resultant optimised dried AM has enhanced structural properties and biochemical stability

  17. Amniotic Fluid or Its Fatty Acids Produce Actions Similar to Diazepam on Lateral Septal Neurons Firing Rate

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    Ana G. Gutiérrez-García

    2013-01-01

    Full Text Available Human amniotic fluid (AF contains eight fatty acids (FATs, and both produce anxiolytic-like effects in adult rats and appetitive responses in human newborns. The medial amygdala and lateral septal nucleus function are related to social behavior, but the action of AF or its FATs in this circuit is known. We obtained 267 single-unit extracellular recordings in Wistar rats treated with vehicle (1 mL, s.c.; n=12, human AF (1 mL, s.c.; n=12, a FAT mixture (1 mL, s.c.; n=13, diazepam (1 mg/kg, i.p.; n=11, and fluoxetine (1 mg/kg, p.o.; n=12. Compared with the vehicle group, the spontaneous septal firing rate in the AF, FAT mixture, and diazepam groups was the lowest and in the fluoxetine group the highest. Cumulative peristimulus histograms indicated that the significant change in septal firing occurred only in the AF and FAT mixture groups and exclusively in those neurons that increased their firing rate during amygdala stimulation. We conclude that human AF and its FATs produce actions comparable to anxiolytic drugs and are able to modify the responsivity of a circuit involved in social behavior, suggesting facilitation of social recognition processes by maternal-fetal fluids.

  18. Amniotic Fluid or Its Fatty Acids Produce Actions Similar to Diazepam on Lateral Septal Neurons Firing Rate

    Science.gov (United States)

    Gutiérrez-García, Ana G.; Vásquez-Hernández, Diana Idania

    2013-01-01

    Human amniotic fluid (AF) contains eight fatty acids (FATs), and both produce anxiolytic-like effects in adult rats and appetitive responses in human newborns. The medial amygdala and lateral septal nucleus function are related to social behavior, but the action of AF or its FATs in this circuit is known. We obtained 267 single-unit extracellular recordings in Wistar rats treated with vehicle (1 mL, s.c.; n = 12), human AF (1 mL, s.c.; n = 12), a FAT mixture (1 mL, s.c.; n = 13), diazepam (1 mg/kg, i.p.; n = 11), and fluoxetine (1 mg/kg, p.o.; n = 12). Compared with the vehicle group, the spontaneous septal firing rate in the AF, FAT mixture, and diazepam groups was the lowest and in the fluoxetine group the highest. Cumulative peristimulus histograms indicated that the significant change in septal firing occurred only in the AF and FAT mixture groups and exclusively in those neurons that increased their firing rate during amygdala stimulation. We conclude that human AF and its FATs produce actions comparable to anxiolytic drugs and are able to modify the responsivity of a circuit involved in social behavior, suggesting facilitation of social recognition processes by maternal-fetal fluids. PMID:23864826

  19. The relationship between the ultrasonographic diagnosis of innocent amniotic band development and pregnancy outcomes.

    Science.gov (United States)

    Wehbeh, H; Fleisher, J; Karimi, A; Mathony, A; Minkoff, H

    1993-04-01

    To determine the importance of amniotic bands diagnosed ultrasonographically and their relationship to pregnancy outcome. We reviewed retrospectively the ultrasonographic records at State University of New York Health Science Center at Brooklyn and Kings County Hospital Center, identifying 25 cases of amniotic band diagnosed during 1986-1990. Cases were compared with 50 controls who had ultrasound evaluation at the same gestational age. Outcomes studied included fetal anomalies, obstetric factors or complications, pregnancy outcome, and maternal factors that may have predisposed to the formation of bands. All cases had unrestricted fetal movement on the index sonogram. No fetal abnormalities were observed in either the case or control series. The groups were not significantly different in terms of maternal risk factors, but significant differences were found for delivery at less than 37 weeks and birth weight less than 2500 g. Ultrasonographic diagnosis of an amniotic band in connection with sonographic findings of normal fetal anatomy may be a risk factor for preterm delivery and low birth weight. No association between amniotic band development and fetal anomalies was observed.

  20. Decreased alpha-fetoprotein in amniotic fluid and maternal serum in diabetic pregnancy

    DEFF Research Database (Denmark)

    Henriques, C U; Damm, P; Tabor, A

    1993-01-01

    OBJECTIVE: To determine a reference level for alpha-fetoprotein (AFP) in the amniotic fluid (AF) in pregnant women with insulin-dependent diabetes mellitus in order to suggest an explanation for the observed decrease in maternal serum AFP. METHODS: Alpha-fetoprotein was measured in AF, maternal...

  1. 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

  2. Use of intra-amniotic urea as a second trimester abortifacient

    Directory of Open Access Journals (Sweden)

    Khare Asha

    1979-01-01

    Full Text Available Seventy cases of mid-trimester pregnancies were terminated by using 200 ml. of 40% intra-amniotic urea, over a 13 month period from 1st August. 1977 to 31st August, 1978. The success rate was 88.7% and the mean induction-abortion interval was 30.7 hours. No major complications were observed.

  3. Evolution of the hypoxia-sensitive cells involved in amniote respiratory reflexes

    NARCIS (Netherlands)

    Hockman, D. (Dorit); A.J. Burns (Alan); Schlosser, G. (Gerhard); Gates, K.P. (Keith P.); Jevans, B. (Benjamin); Mongera, A. (Alessandro); S. Fisher (Shannon); Unlu, G. (Gokhan); Knapik, E.W. (Ela W.); Kaufman, C.K. (Charles K.); Mosimann, C. (Christian); L.I. Zon (Leonard); Lancman, J.J. (Joseph J.); Dong, P.D.S. (P. Duc S.); Lickert, H. (Heiko); Tucker, A.S. (Abigail S.); Baker, C.V.H. (Clare V. H.)

    2017-01-01

    textabstractThe evolutionary origins of the hypoxia-sensitive cells that trigger amniote respiratory reflexes – carotid body glomus cells, and ‘pulmonary neuroendocrine cells’ (PNECs) -are obscure. Homology has been proposed between glomus cells, which are neural crest-derived, and the

  4. Assessment of amniotic and polyurethane membrane dressings in the treatment of burns.

    Science.gov (United States)

    Adly, O A; Moghazy, A M; Abbas, A H; Ellabban, A M; Ali, O S; Mohamed, B A

    2010-08-01

    As allograft and xenografts are not available in Islamic countries, amniotic membrane seems to be an effective alternative in the management of deep burns. Its proven bioactivities and modest price suggest that it might be superior to synthetic dressings. Forty-six patients were enrolled in this randomized, controlled clinical trial conducted in the Burn Unit at Suez Canal University Hospital, Ismailia, Egypt. All age groups and both gender were included in the study. Only patients with less than 50% total body surface area burned were included, thus minimizing the dropouts in both groups. All were either second or third degree. These patients were randomly assigned either to group I: amniotic membrane (Biomembrane) dressing, or group II: polyurethane membrane (Tegaderm) dressing. Those in group I demonstrated a significantly lower rate of infection and required less frequent dressing changes than those in group II. They also sustained less electrolyte and albumin loss. The rate of healing in the amniotic membrane group was significantly faster than in the polyurethane group. Furthermore, pain was significantly less when Biomembrane was used. Based on these findings, we recommend the use of lyophilized gamma-irradiated amniotic membrane as an effective alternative for allograft and xenografts in Islamic countries and the Jewish population.

  5. 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.

  6. Bacterial aetiological agents of intra-amniotic infections and preterm birth in pregnant women

    Directory of Open Access Journals (Sweden)

    George Louis Mendz

    2013-10-01

    Full Text Available Infection-related preterm birth is a leading cause of infant mortality and morbidity; knowledge of bacterial populations invading the amniotic cavity and the routes of invasion is required to make progress in the prevention of preterm birth. Significant advances have been made in understanding bacterial communities in the vagina, but much less studied are intra-uterine bacterial populations during pregnancy. A systematic review of data published on the intra-uterine microbiome was performed; molecular information and summaries of species found in healthy individuals and in women with diagnosed infections served to construct a database and to analyse results to date. Thirteen studies fulfilled the review's inclusion criteria. The data of various investigations were collated, organised and re-analysed to achieve a more comprehensive understanding of microbial populations in the intra-amniotic space. The most common intra-amniotic bacterial taxa were species that can colonise the vagina in health and disease; there were others associated with the habitats of the mouth, gastrointestinal tract and respiratory tract. The results suggest a central role for the ascending route of infections during pregnancy, and points to a possible secondary contribution via haematogenous invasion of the intra-amniotic space. The census of the intra-uterine microbiome awaits completion.

  7. Clinical research of tear film stability after pterygium excision with amniotic membrane transplantation

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    Guo-Ying Ming

    2014-08-01

    Full Text Available AIM: To assess changes of tear film function in patients after pterygium excision combined with amniotic membrane transplantation.METHODS:Totally 126 patients with pterygium excision with amniotic membrane transplantation from January 2011 to November 2013 were entered in the study. The tear breakup time(BUT, the Schirmer I test(SⅠtand tear ferning test(TFTwere elevated in the patients before and after pterygium excision combined with amniotic membrane transplantation. The examnation times were 1d before surgey, 1wk, 1, 2mo after surgery. Operation eyes were studied group, while opposite healthy eyes as control group.RESULTS: Compared with the control group, BUT and TFT were significantly different in the eyes with pterygium(PP>0.05. The results of BUT and TFT at 1mo after surgery in study group were significantly better than 1wk(PP>0.05; The tear film stability in the study group at 1wk after surgery was still inferior to the control group(PP all>0.05. SⅠt results did not differ between the different examination times(P>0.05.CONCLUSION:Tear film stability was broken in the eyes with pterygium. Pterygium excision combined with amniotic membrane transplantation can obviously restore the tear film function into normal state, and the tear film function could reach steady-state 1mo after surgery.

  8. Amniotic Fluid Stem Cells and Their Application in Cell-Based Tissue Regeneration

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    Mohamadreza Baghaban Eslaminejad

    2012-01-01

    Full Text Available Advances in stem cell biotechnology hold great promise in the field of tissue engineering andregenerative medicine. Of interest are marrow mesenchymal stem cells (MSCs, embryonic stemcells (ESCs, and induced pluripotent stem cells (iPSCs. In addition, amniotic fluid stem cells (AFSCshave attracted attention as a viable choice following the search for an alternative stem cellsource. Investigators are interested in these cells because they come from the amniotic fluid that isroutinely discarded after birth. There have been multiple investigations conducted worldwide in anattempt to better understand AF-SCs in terms of their potential use in regenerative medicine. In thisreview we give a brief introduction of amniotic fluid followed by a description of the cells presentwithin this fluid. Their history related to stem cell discovery in the amniotic fluid as well as themain characteristics of AF-SCs are discussed. Finally, we elaborate on the potential for these cellsto promote regeneration of various tissue defects, including fetal tissue, the nervous system, heart,lungs, kidneys, bones, and cartilage.

  9. Amniotic membrane transplantation in corneal melting after anterior lamellar keratoplasty assisted by femtosecond laser in children.

    Science.gov (United States)

    Buzzonetti, Luca; Petrocelli, Gianni; Valente, Paola

    2012-01-01

    To evaluate the efficacy of amniotic membrane transplantation in lamella melting after anterior lamellar keratoplasty (ALK) assisted by femtosecond laser in a pediatric patient. An 11-year-old girl with progressive keratoconus underwent ALK assisted by femtosecond laser on the right eye. The surgical procedure was performed under general anesthesia. The 60-KHz IntraLase femtosecond laser (Abbott Medical Optics) created both the donor (thickness 350 µm; diameter 8.1 mm; side cut 70°) and recipient (thickness 260 µm; diameter 8.0 mm; side cut 70°) lamellae. The recipient lamella was then gently removed and donor was fitted into place and sutured using 4 interrupted sutures added to a running suture in nylon 10-0. The surgery was uneventful. Two months after surgery, a lamella melting was observed. One month after topical steroid treatment, amniotic membrane transplantation was performed. When lamella melting was observed, the best-corrected visual acuity (BCVA) was 0.1 and 0.5 with foramen and no inflammatory reaction in the anterior chamber was reported. Eleven months after amniotic membrane transplantation, BCVA was 0.9 and no visual acuity increase was recorded with foramen. Our findings show that amniotic membrane transplantation could be considered in lamella melting after ALK assisted by femtosecond laser in children.

  10. ALPHA-FETOPROTEIN IN FETAL SERUM, AMNIOTIC-FLUID, AND MATERNAL SERUM

    NARCIS (Netherlands)

    VANLITH, JMM; BEEKHUIS, [No Value; VANLOON, AJ; MANTINGH, A; DEWOLF, BTHM; BREED, ASPM

    In order to gain more insight into the association between alpha-fetoprotein (AFP) and fetal chromosomal disorders, especially Down's syndrome, we measured AFP in fetal serum, amniotic fluid, and maternal serum at cordocentesis. We compared the concentration and gradient of AFP in these three

  11. Neuroinflammation and structural injury of the fetal ovine brain following intra-amniotic Candida albicans exposure

    NARCIS (Netherlands)

    Ophelders, D.R.; Gussenhoven, R.; Lammens, M.; Kusters, B.; Kemp, M.W.; Newnham, J.P.; Payne, M.S.; Kallapur, S.G.; Jobe, A.H.; Zimmermann, L.J.; Kramer, B.W.; Wolfs, T.G.

    2016-01-01

    BACKGROUND: Intra-amniotic Candida albicans (C. Albicans) infection is associated with preterm birth and high morbidity and mortality rates. Survivors are prone to adverse neurodevelopmental outcomes. The mechanisms leading to these adverse neonatal brain outcomes remain largely unknown. To better

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

    Directory of Open Access Journals (Sweden)

    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

  13. Levels of Pesticides and Their Metabolites in Wistar Rat Amniotic Fluids and Maternal Urine upon Gestational Exposure

    Science.gov (United States)

    Bossi, Rossana; Vinggaard, Anne Marie; Taxvig, Camilla; Boberg, Julie; Bonefeld-Jørgensen, Eva Cecilie

    2013-01-01

    Concentrations of pesticides and selected metabolites in rat urine and amniotic fluid were determined as biomarker upon oral administration of Wistar rats to two pesticide mixtures consisting of three to five pesticides (bitertanol, propiconazole, cypermethrin, malathion, and terbuthylazine). The pesticides and their metabolites were found in rat amniotic fluid and urine, generally in dose-response concentrations in relation to dosage. The measurement of the substances in the amniotic fluid indicated that the fetus was exposed to the pesticides as well as their metabolites. Moreover, the pesticides detected in urine demonstrated the exposure as well as the ability of the rat to excrete these compounds. PMID:23736656

  14. Levels of Pesticides and Their Metabolites in Wistar Rat Amniotic Fluids and Maternal Urine upon Gestational Exposure

    DEFF Research Database (Denmark)

    Bossi, Rossana; Vinggaard, Anne Marie; Taxvig, Camilla

    2013-01-01

    Concentrations of pesticides and selected metabolites in rat urine and amniotic fluid were determined as biomarker upon oral administration of Wistar rats to two pesticide mixtures consisting of three to five pesticides (bitertanol, propiconazole, cypermethrin, malathion, and terbuthylazine......). The pesticides and their metabolites were found in rat amniotic fluid and urine, generally in dose-response concentrations in relation to dosage. The measurement of the substances in the amniotic fluid indicated that the fetus was exposed to the pesticides as well as their metabolites. Moreover, the pesticides...

  15. Influence of maternal age, gestational age and fetal gender on expression of immune mediators in amniotic fluid

    OpenAIRE

    Weissenbacher Tobias; Laubender Rüdiger P; Witkin Steven S; Gingelmaier Andrea; Schiessl Barbara; Kainer Franziskus; Friese Klaus; Jeschke Udo; Dian Darius; Karl Katrin

    2012-01-01

    Abstract Background Variations in cytokine and immune mediator expression patterns in amniotic fluid due to gestational age, maternal age and fetal gender were investigated. Findings Amniotic fluid samples were obtained from 192 women, 82 with a mid-trimester amniocentesis (median gestational age 17 weeks) and 110 with a caesarean section not in labor (median gestational age 39 weeks). Amniotic fluid was screened by commercial ELISAs for the TH1/TH2/TH17 cytokines and immune mediators IL-1 be...

  16. [Multilayer amniotic membrane transplantation for treatment of necrotizing herpes simplex stromal keratitis].

    Science.gov (United States)

    Shi, Wei-yun; Chen, Min; Wang, Fu-hua; Zhao, Jing; Ma, Lin; Xie, Li-xin

    2005-12-01

    To evaluate the efficacy of multilayer amniotic membrane transplantation (AMT) combined with antivirus and corticosteroid drug to treat necrotizing herpes simplex stromal keratitis. Thirteen patients (13 eyes) of necrotizing stromal keratitis were referred to Shandong Eye Institute and Qingdao Eye Hospital between January 2003 and April 2004. The course of disease was 3 - 22 months (mean 15 months). Corneal inflammation persisted and corneal ulcer progressed despite topical and systemic antiviral treatment for over 1 weeks. Multilayer amniotic membrane transplantation was performed after excluding of bacterial and fungal infection by microbiologic studies including smears and cultures of necrotic corneal tissue and confocal microscope. Topical and systemic antiviral medications were given with adjuvant corticosteroid eyedrops postoperatively. We investigated the healing of corneal ulcer and improvement of stromal edema with slit lamp biomicroscope, the integrity of corneal defect with fluorescein staining, the migration of healthy corneal epithelial cells and transformation of amniotic membrane with confocal microscopy. All patients were followed up for 3 - 13 months (mean 10 months). Corneal ulcer healed within 1 - 3 weeks postoperatively with negative fluorescein staining. Corneal stromal edema faded away within 1 month. Superficial amniotic membrane patches dissolved or shed on postoperative day 7 - 10, while the deeper grafts were adhered into the ulcer and fused with the surrounding fibroblasts. One of these grafts remained in situ more than 3 months. Confocal microscope examination indicated flat epithelial progenitor cells on the surface of residual amniotic membrane. Corneal transparence was achieved in 7 eyes, macula in 4 eyes and leucoma in 2 eyes 3 months after the operation. No recurrence of necrotizing stromal keratitis was occurred in 13 patients during the follow-up period. Multilayer AMT combined with antivirus and corticosteroid treatment is an

  17. Membrana amniótica como alternativa de tratamiento en superficie ocular Amniotic membrane as a therapeutic option for the ocular surface

    Directory of Open Access Journals (Sweden)

    Keyly Fernández García

    2012-12-01

    Full Text Available Se realizó una revisión bibliográfica con el objetivo de conocer la utilidad de la membrana amniótica como alternativa de tratamiento en la superficie ocular. Son abordados tópicos como las diferentes formas de obtención, preparación y conservación de la misma, sus mecanismos de acción y aplicaciones. Se consultó una bibliografía que abarca un periodo de varios años para conocer los resultados publicados sobre el trasplante de membrana amniótica humana en la superficie ocular.A literature review was made to learn about the usefulness of the amniotic membrane as a therapeutic option for the ocular surface. The different ways for obtaining, preparing, and conserving this membrane, its mechanism of action and its applications were also addressed. Literature covering several years was reviewed in order to be acquainted with the published results of the human amniotic membrane transplantation on the ocular surface.

  18. Phytoestrogens in Human Pregnancy

    Directory of Open Access Journals (Sweden)

    John Jarrell

    2012-01-01

    Full Text Available Background. The hormonal milieu associated with pregnancy has become a focus of interest owing to potential links with the developmental origins of health and disease. Phytoestrogens are hormonally active plant-derived chemicals that may have an impact on human reproductive processes. However, developmental exposure to phytoestrogens has not been well characterized and thus our objective was to quantify phytoestrogen exposure during pregnancy and lactation. Methods. Women in the second trimester of pregnancy entered the study during counseling for prenatal genetic information. Women who had an indication for a genetic amniocentesis on the basis of late maternal age were approached for inclusion. They completed an environmental questionnaire; a sample of amniotic fluid was collected for karyotype, blood was collected from women during pregnancy and at birth, from the umbilical cord and breast milk. Samples were tested for the presence of daidzein and genistein by GC Mass Spectroscopy. Findings. Phytoestrogens are commonly found in pregnant women’s serum and amniotic fluid during pregnancy. There is a sex difference in the concentrations with higher levels in amniotic fluid containing female fetuses. This difference was not present in maternal serum. Soy ingestion increases amniotic fluid phytoestrogen concentrations in female and male fetuses. The presence and concentrations of phytoestrogens did not differ in relation to common pregnancy complications or preexisting infertility.

  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

    Proteins found in amniotic fluid are mainly serum proteins, probably of maternal origin. About 5% of the total protein concentration has the potential of being fetal or decidual in origin. Only a few of these proteins have been isolated and characterised. In order to describe the foetal...... and decidual components in amniotic fluid more extensively, a polyspecific antiserum to amniotic fluid at weeks 16-18 of gestation was raised. Specificities in the antiserum to serum proteins were removed by adsorption. Several proteins of non-serum protein origin reacted with the antiserum. Three...... 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. STUDY OF BIOCHEMICAL PARAMETERS IN AMNIOTIC FLUID FOR ASSESSMENT OF FOETAL MATURITY IN CASES OF NORMAL PREGNANCY

    Directory of Open Access Journals (Sweden)

    Leela

    2015-11-01

    Full Text Available Assessment of foetal maturity had been proven of value in evaluating the foetal condition. Accurate assessment of foetal maturity is essential for the proper timing of delivery in various risk pregnancies. Amniotic Fluid analysis for foetal maturity had been of proven value. In the present study, study of biochemical parameters in amniotic fluid in respect of Creatinine, Uric Acid, Urea, Total Proteins, and Electrolytes i.e. Sodium, Potassium and Chloride has been done, along with Serum Electrolytes. Standard methodologies were adopted. The observations in the present study correlated with the works of Chadick et al and Pitkin and Zwirek. The levels of Creatinine, Uric Acid and Urea in Amniotic Fluid showed elevation, while Total Proteins and Serum Sodium showed a decline, as gestation progressed. The Serum and Amniotic Fluid Potassium and Chloride levels remain almost constant throughout the pregnancy. Thus, it is observed that the use of multiple parameters is desirable for accurate assessment of foetal maturity.

  1. Evolution of hard proteins in the sauropsid integument in relation to the cornification of skin derivatives in amniotes

    National Research Council Canada - National Science Library

    Alibardi, Lorenzo; Dalla Valle, Luisa; Nardi, Alessia; Toni, Mattia

    2009-01-01

    Hard skin appendages in amniotes comprise scales, feathers and hairs. The cell organization of these appendages probably derived from the localization of specialized areas of dermal-epidermal interaction in the integument...

  2. Amniotic fluid insulin and C-peptide as predictive markers for fetal macrosomia, birth injuries, and delivery complications?

    OpenAIRE

    Stein, Roland Gregor; Meinusch, Malgorzata; Diessner, Joachim; Dietl, Johannes; Hönig, Arnd; Zollner, Ursula

    2014-01-01

    Background Gestational diabetes mellitus (GDM) occurs in 3–5% of all pregnancies. GDM increases both maternal and fetal risks, causes fetal macrosomia, and hence increases the rates of caesarean sections and delivery complications such as shoulder dystocia. An early predictive marker and consequent early treatment could be beneficial, so amniotic fluid insulin and C-peptide have been examined in several studies. Increased amniotic fluid insulin in early amniocentesis between the 14th and 20th...

  3. Clinical observation of corneal lamellar debridement combined with sutureless amniotic membrane transplantation for the treatment of superficial fungal keratitis

    Directory of Open Access Journals (Sweden)

    Huang Zhang

    2014-09-01

    Full Text Available AIM:To evaluate the clinical efficacy of corneal lamellar debridement combined with sutureless amniotic membrane transplantation for the treatment of superficial fungal keratitis.METHODS:Totally 22 cases(22 eyeswith superficial fungal keratitis were referred to our hospital from April 2012 to October 2013. The patients with persistent cornea ulcer after treatment of local and systemic antifungal drugs underwent corneal lamellar debridement combined with sutureless amniotic membrane transplantation, and the recipient bed was covered with an amniotic membrane using fibrin sealant during the operation. All patients were still given topical antifungal therapy for 1-2mo after operation. The followed-up time was 3mo or above. We observed the corneal healing and amniotic membrane adhesion by split lamp microscope, and investigated the transformation of amniotic membrane and fungal infection recurrence with confocal microscope. RESULTS: Corneal edema and anterior chamber reaction of 21 patients disappeared gradually, and no amniotic membrane graft dissolved and shed off within 1-2wk postoperatively. Two weeks after operation, the graft integrated into the corneal and the corneal wounds' thickness increased gradually, the corneal epithelium reconstructed and corneas became clear. Four weeks after operation, the corneal scarring developed gradually and fluorescence staining was negative. Nineteen cases' amniotic membranes that adhered with the cornea dissolved 4wk after operation. There were different degrees of corneal nebula or macula remained 3mo postoperatively. All patients' vision improved in varying degrees, except in 1 case with fungal keratitis who had been cured by lamellar keratoplasty.CONCLUSION:Corneal lamellar debridement combined with sutureless amniotic membrane transplantation can effectively remove the foci of inflammation, improve the local efficacy, shorten the operation time, relieve the postoperative reaction, and promote cornea

  4. The Role of the Multiple Banded Antigen of Ureaplasma parvum in Intra-Amniotic Infection: Major Virulence Factor or Decoy?

    OpenAIRE

    Dando, Samantha J.; Nitsos, Ilias; Kallapur, Suhas G.; Newnham, John P.; Polglase, Graeme R.; Pillow, J. Jane; Jobe, Alan H.; Timms, Peter; Knox, Christine L.

    2012-01-01

    The multiple banded antigen (MBA) is a predicted virulence factor of Ureaplasma species. Antigenic variation of the MBA is a potential mechanism by which ureaplasmas avoid immune recognition and cause chronic infections of the upper genital tract of pregnant women. We tested whether the MBA is involved in the pathogenesis of intra-amniotic infection and chorioamnionitis by injecting virulent or avirulent-derived ureaplasma clones (expressing single MBA variants) into the amniotic fluid of pre...

  5. Amniotic membrane as part of a skin substitute for full-thickness wounds: an experimental evaluation in a porcine model.

    Science.gov (United States)

    Loeffelbein, Denys J; Baumann, Claudia; Stoeckelhuber, Mechthild; Hasler, Rafael; Mücke, Thomas; Steinsträßer, Lars; Drecoll, Enken; Wolff, Klaus-Dietrich; Kesting, Marco R

    2012-07-01

    We evaluated the use of human amniotic membrane (HAM) as a graft material for the treatment of iatrogenic full-thickness (FT) skin wounds in a porcine model with a view to reducing donor site morbidity in free flap transfer. Forty experimental FT-wounds were covered with an autologous split-thickness skin graft (STSG) alone or in combination with a mono- or multilayer HAM or Integra(®). Untreated wounds served as controls. Clinical evaluation and biopsy-sampling for histological and immunohistochemical staining with von-Willebrand-factor (vWF) antibody, laminin antibody, Ki-67 antibody, and smooth muscle actin (αSMA) antibody were performed on days 5, 7, 10, 20, 40, and 60 after surgical intervention. Considerable disparities in the estimated criteria were observed between the various treatment groups of the FT-wounds. The use of HAM was found to have an accelerating impact on re-epithelialization. The multilayered amnion membrane showed better results than the Integra(®) and monolayer technique in terms of contraction rate, inflammation, and scarring and seemed useful as a dermal substitute in FT-wounds giving comparable results to STSG coverage alone. This study demonstrates the successful application of HAM as part of a skin substitute in FT-wounds in minipigs. The results offer promise as a simple and effective technique for the application of multilayer HAM in iatrogenic human skin defects and the acceleration of wound healing. Copyright © 2012 Wiley Periodicals, Inc.

  6. Histochemical, Biochemical and Cell Biological aspects of tail regeneration in lizard, an amniote model for studies on tissue regeneration.

    Science.gov (United States)

    Alibardi, Lorenzo

    2014-01-01

    and inflammatory course, an inspiring model for understanding failure of tissue regeneration in higher vertebrates and humans. The participation of 5-Bromo-deoxyuridine (5BrdU) long retention cells, indicated as putative stem cells, for the following regeneration is analyzed and it shows that different tissue sites of the original tail contain putative stem cells that are likely activated from the wounding signal. In particular, the permanence of stem cells in the central canal of the spinal cord can explain the limited but important neurogenesis present in the caudal but also in the lumbar-thoracic spinal cord. In the latter, the limited number of glial and neurons regenerated is however sufficient to recover some limited hind limb movement after injury or spinal transection. Finally, the presence of stem cells in the spinal cord, in the regenerative blastema and skin allow to use these organs as a source of cells for studies on gene activation during cell differentiation in the new spinal cord, tail and in the epidermis. The above information form the basic knowledge for the future molecular studies on the specific gene activation capable to determine tail regeneration in lizards, and more in general genes involved in the reactivation of regeneration process in amniotes and humans. Copyright © 2013 Elsevier GmbH. All rights reserved.

  7. Prenatal imaging of amniotic band sequence: utility and role of fetal MRI as an adjunct to prenatal US

    Energy Technology Data Exchange (ETDEWEB)

    Neuman, Jeremy [Cincinnati Children' s Hospital Medical Center, Department of Radiology, Cincinnati, OH (United States); Staten Island University Hospital, Department of Radiology, Staten Island, NY (United States); Calvo-Garcia, Maria A.; Kline-Fath, Beth M.; Bitters, Constance; Merrow, Arnold C.; Guimaraes, Carolina V.A. [Cincinnati Children' s Hospital Medical Center, Department of Radiology, Cincinnati, OH (United States); Lim, Foong-Yen [Cincinnati Children' s Hospital Medical Center, Department of Fetal Surgery, Cincinnati, OH (United States)

    2012-05-15

    Amniotic band sequence and its US manifestations have been well-described. There is little information, however, regarding the accuracy and utility of fetal MRI. To describe the MRI findings in amniotic band sequence and to compare the diagnostic accuracy of MRI and US. Prenatal MRI and US studies were retrospectively reviewed in 14 consecutive pregnancies with confirmed amniotic band sequence. Both studies were evaluated for amniotic band visualization, body part affected, type of deformity, umbilical cord involvement and vascular abnormality. Amniotic bands were confidently identified with MRI in 8 fetuses (57%), suggested with MRI in 3 fetuses (21%) and confidently seen by US in 13 fetuses (93%). Neither modality detected surgically proven bands on one fetus. Both techniques were equally able to define the body part affected and the type of deformity. At least one limb abnormality was visualized in all cases and truncal involvement was present in two cases. Cord involvement was identified in seven cases, with one case detected only by MRI. Fetal MRI is able to visualize amniotic bands and their secondary manifestations and could be complementary to prenatal US when fetal surgery is contemplated. (orig.)

  8. Are amniotic fluid neutrophils in women with intraamniotic infection and/or inflammation of fetal or maternal origin?

    Science.gov (United States)

    Gomez-Lopez, Nardhy; Romero, Roberto; Xu, Yi; Leng, Yaozhu; Garcia-Flores, Valeria; Miller, Derek; Jacques, Suzanne M; Hassan, Sonia S; Faro, Jonathan; Alsamsam, Adham; Alhousseini, Ali; Gomez-Roberts, Hunter; Panaitescu, Bogdan; Yeo, Lami; Maymon, Eli

    2017-09-28

    Neutrophils are the most abundant white blood cells found in the amniotic cavity of women with intraamniotic infection and/or inflammation. The current belief is that these neutrophils are of fetal origin. However, abundant neutrophils have been found in the amniotic fluid of women with a severe acute maternal inflammatory response but without a fetal inflammatory response in the placenta, suggesting that these innate immune cells can also be of maternal origin or a mixture of both fetal and maternal neutrophils. We sought to investigate the origin of amniotic fluid neutrophils from women with intraamniotic infection and/or inflammation, and to correlate these findings with acute histologic maternal and fetal inflammatory responses in the placenta. Amniotic fluid was collected from 15 women with suspected intraamniotic infection and/or inflammation (positive microbiological cultures and/or interleukin-6 concentrations ≥2.6 ng/mL). Amniotic fluid neutrophils were purified by fluorescence-activated cell sorting, DNA was extracted, and DNA fingerprinting was performed. DNA fingerprinting was also performed in the umbilical cord and maternal blood DNA. Fluorescence in situ hybridization was assayed in women with male neonates. Blinded placental histopathological evaluations were conducted. First, DNA fingerprinting revealed that 42.8% (6/14) of women who underwent a single amniocentesis had mostly fetal neutrophils in the amniotic fluid. Second, DNA fingerprinting showed that 35.7% (5/14) of the women who underwent a single amniocentesis had predominantly maternal neutrophils in the amniotic fluid. Third, DNA fingerprinting indicated that 21.4% (3/14) of the women who underwent a single amniocentesis had an evident mixture of fetal and maternal neutrophils in the amniotic fluid. Fourth, DNA fingerprinting revealed that a woman who underwent 2 amniocenteses (patient 15) had fetal neutrophils first, and as infection progressed, abundant maternal neutrophils invaded the

  9. Treatment of ligneous conjunctivitis with amniotic membrane transplantation and topical cyclosporine

    Directory of Open Access Journals (Sweden)

    Ozlem Yalcin Tok

    2012-01-01

    Full Text Available Ligneous conjunctivitis (LC is a rare form of bilateral chronic recurrent disease in which thick membranes form on the palpebral conjunctiva and other mucosal sites. We report the clinical features and describe the management of two cases. Case 1 was an 8-month-old patient with bilateral membranous conjunctivitis. Case 2 was a 5-year-old patient with unilateral membranous conjunctivitis, esotropia, mechanical ptosis and complicated cataract, and had been treated with a number of medications. Histological investigation of the membrane in both cases showed LC. Treatments with amniotic membrane transplantation and institution of topical cyclosporine have shown good response. There has been complete resolution of the membranes with no recurrence at the end of 40- and 28-month follow-ups, respectively. No treatment related side effects were seen. Thus, it appears that amniotic membrane transplantation and topical cyclosporine are effective alternatives for the treatment of LC.

  10. Predicting fetal lung maturity by visual assessment of amniotic fluid turbidity: comparison with fluorescence polarization assay.

    Science.gov (United States)

    Adair, C D; Sanchez-Ramos, L; McDyer, D L; Gaudier, F L; Del Valle, G O; Delke, I

    1995-10-01

    We prospectively studied 159 patients having clinically indicated amniocentesis. Amniotic fluid (3 to 5 mL) was placed in a nonheparinized glass tube. This sample was then classified as turbid (indicating maturity) or clear (indicating immaturity) on the basis of a single examiner's ability to read newspaper print through the glass tube. These results were then compared with fluorescence polarization values for the same sample. A value of 70 mg/g was considered positive evidence of fetal lung maturity. By study criteria, 62 samples (39%) indicated immaturity and 97 (61%) indicated maturity. Turbidity correctly identified 89 samples that produced fluorescence polarization values of at least 70 mg/g. Turbidity as a predictor of fetal lung maturity when compared with fluorescence polarization assay has a 91% positive and 87% negative predictive value. Visual inspection of amniotic fluid may be of value in areas where sophisticated methods are unavailable.

  11. Germline development in amniotes: A paradigm shift in primordial germ cell specification

    Science.gov (United States)

    2016-01-01

    In the field of germline development in amniote vertebrates, primordial germ cell (PGC) specification in birds and reptiles remains controversial. Avians are believed to adopt a predetermination or maternal specification mode of PGC formation, contrary to an inductive mode employed by mammals and, supposedly, reptiles. Here, we revisit and review some key aspects of PGC development that channelled the current subdivision, and challenge the position of birds and reptiles as well as the ‘binary’ evolutionary model of PGC development in vertebrates. We propose an alternative view on PGC specification where germ plasm plays a role in laying the foundation for the formation of PGC precursors (pPGC), but not necessarily of PGCs. Moreover, inductive mechanisms may be necessary for the transition from pPGCs to PGCs. Within this framework, the implementation of data from birds and reptiles could provide new insights on the evolution of PGC specification in amniotes. PMID:27273724

  12. Treatment of ligneous conjunctivitis with amniotic membrane transplantation and topical cyclosporine.

    Science.gov (United States)

    Tok, Ozlem Yalcin; Kocaoglu, Fatma Akbas; Tok, Levent; Burcu, Ayse; Ornek, Firdevs

    2012-01-01

    Ligneous conjunctivitis (LC) is a rare form of bilateral chronic recurrent disease in which thick membranes form on the palpebral conjunctiva and other mucosal sites. We report the clinical features and describe the management of two cases. Case 1 was an 8-month-old patient with bilateral membranous conjunctivitis. Case 2 was a 5-year-old patient with unilateral membranous conjunctivitis, esotropia, mechanical ptosis and complicated cataract, and had been treated with a number of medications. Histological investigation of the membrane in both cases showed LC. Treatments with amniotic membrane transplantation and institution of topical cyclosporine have shown good response. There has been complete resolution of the membranes with no recurrence at the end of 40- and 28-month follow-ups, respectively. No treatment related side effects were seen. Thus, it appears that amniotic membrane transplantation and topical cyclosporine are effective alternatives for the treatment of LC.

  13. Treatment of ligneous conjunctivitis with amniotic membrane transplantation and topical cyclosporine

    Science.gov (United States)

    Tok, Ozlem Yalcin; Kocaoglu, Fatma Akbas; Tok, Levent; Burcu, Ayse; Ornek, Firdevs

    2012-01-01

    Ligneous conjunctivitis (LC) is a rare form of bilateral chronic recurrent disease in which thick membranes form on the palpebral conjunctiva and other mucosal sites. We report the clinical features and describe the management of two cases. Case 1 was an 8-month-old patient with bilateral membranous conjunctivitis. Case 2 was a 5-year-old patient with unilateral membranous conjunctivitis, esotropia, mechanical ptosis and complicated cataract, and had been treated with a number of medications. Histological investigation of the membrane in both cases showed LC. Treatments with amniotic membrane transplantation and institution of topical cyclosporine have shown good response. There has been complete resolution of the membranes with no recurrence at the end of 40- and 28-month follow-ups, respectively. No treatment related side effects were seen. Thus, it appears that amniotic membrane transplantation and topical cyclosporine are effective alternatives for the treatment of LC. PMID:23202401

  14. [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.

  15. Thoraco-amniotic shunting for fetal pleural effusion--a case series.

    LENUS (Irish Health Repository)

    Walsh, J

    2011-11-15

    Fetal pleural effusion is a rare occurrence, with an incidence of 1 per 10-15,000 pregnancies. The prognosis is related to the underlying cause and is often poor. There is increasing evidence that in utero therapy with thoraco-amniotic shunting improves prognosis by allowing lung expansion thereby preventing hydrops and pulmonary hypoplasia. This is a review of all cases of fetal pleural effusion managed over an eight year period the National Maternity Hospital Dublin. Over the nine year period there were 21 cases of fetal pleural effusion giving an overall incidence of 1 per 9281 deliveries. Of these, 15 underwent thoraco-amniotic shunting. There were associated anomalies diagnosed in 5 (33%) of cases. The overall survival in our cohort was 53%. The presence of hydrops was a poor prognostic factor, with survival in cases with hydrops of 33% (3\\/9) compared to 83% (5\\/6) in those cases without associated hydrops.

  16. 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...

  17. 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...

  18. AMNIOTIC MEMBRANE TRANSPLANTATION IN OCULAR SURFACE DISORDERS – A CASE SERIES

    Directory of Open Access Journals (Sweden)

    Umesh

    2016-03-01

    Full Text Available ABSTRACT The cornea, conjunctiva and the limbus comprise the tissues at the ocular surface. All of them are covered by stratified, squamous, non-keratinising epithelium and a stable tear film. The ocular surface health is ensured by intimate relationship between ocular surface epithelia and the preocular tear film. It is essential to establish accurate diagnosis for appropriate management of complex ocular surface disorders. Management has improved with introduction of the limbal stem cell concept and amniotic membrane transplantation.

  19. Comparison of Amniotic Fluid Cytokine Levels in Postterm and Term Pregnancy: a Prospective Study.

    Science.gov (United States)

    Korkmaz, Vakkas; Cekmez, Yasemin; Surer, Hatice; Deveci, Canan D; Ocal, Fatma D; Gezer, Murad; Kurdoglu, Zehra; Ergun, Yusuf; Kucukozkan, Tuncay

    2017-02-01

    To evaluate amniotic fluid pro- and anti-inflammatory cytokine levels in women with postterm and term pregnancies in labor and not in labor. The study involved three groups: postterm (Group 1, n = 29), term in labor (Group 2, n = 28), and control (Group 3, n = 30). All groups were compared with respect to age, gravidity, parity, obstetric history, gestation week, cervical dilatation and effacement, maternal serum C-reactive protein and white cell count, amniotic interleukin 4, 6, and 10 levels, birthweight, and cord blood pH. The amniotic fluid interleukin 10 level was 24.4 ± 8.8 pg/mL in the postterm group, 13.5 ± 5.1 pg/mL in the term in labor group, and 19.8 ± 5.4 pg/mL in the control group (p postterm group, 38.2 ± 29.2 pg/mL in the term in labor group, and 81.9 ± 68.4 pg/mL in the control group (p = 0.002). The amniotic fluid interleukin 6 level was 329 ± 135.1 pg/mL in the postterm group, 252.8 ± 138.7 pg/mL in the term in labor group, and 227.9 ± 114.4 pg/mL in the control group (p = 0.02). There was a positive correlation between gestational age and IL-10 levels (p postterm pregnancy and they decreased with active labor.

  20. Postnatal amniotic fluid intake reduces gut inflammatory responses and necrotizing enterocolitis in preterm neonates

    DEFF Research Database (Denmark)

    Siggers, Jayda; Østergaard, Mette V.; Siggers, Richard H.

    2013-01-01

    Preterm neonates are susceptible to gastrointestinal disorders such as necrotizing enterocolitis (NEC). Maternal milk and colostrum protects against NEC via growth promoting, immunomodulatory, and antimicrobial factors. The fetal enteral diet amniotic fluid (AF), contains similar components, and we.......9 and 7.7 compared with 17.3, P inflammation-related genes (e.g., TNF-α, IL-1α, IL-6, NOS) were downregulated, relative to FORM pigs with NEC. Anti...

  1. Measurement of amniotic fluid steroids of midgestation via LC-MS/MS.

    Science.gov (United States)

    Fahlbusch, Fabian B; Heussner, Kirsten; Schmid, Matthias; Schild, Ralf; Ruebner, Matthias; Huebner, Hanna; Rascher, Wolfgang; Doerr, Helmuth-Guenther; Rauh, Manfred

    2015-08-01

    Analysis of steroids by mass spectrometry (MS) has evolved into a reliable tool for the simultaneous detection of multiple steroids. As amniotic fluid (AF) and fetal serum composition of early pregnancy are closely related, the analysis of AF can yield information on the physiological status of the developing fetus. We evaluated the use of liquid-chromatography tandem mass spectrometry (LC-MS/MS) for AF steroid analysis, including the analysis of its sensitivity and accuracy for gender verification in healthy subjects. AF of 78 male and 94 female healthy newborns was analyzed by LC-MS/MS at 16 weeks of gestation. The levels of androstenedione, corticosterone, cortisol, cortisone, deoxycorticosterone, 11-deoxycortisol, dehydroepiandrosterone (DHEA), dehydroepiandrosterone-sulfate (DHEA-S), 17-hydroxyprogesterone, progesterone (17-OHP) and testosterone were measured. Steroid levels were compared to RIA and GC-MS levels of midgestation from the literature. Cross-validated logistic regression was used to obtain statistical predictions of gender at birth from testosterone and the above steroids. LC-MS/MS analysis of AF steroids yielded comparable results with published GC-MS data. Gender specific differences were found for androstenedione and testosterone concentrations with higher levels in the male fetus. In contrast to published RIA data no gender specific differences were observed for 17-hydroxyprogesterone and dehydroepiandrosterone AF concentrations. Testosterone concentrations yielded highly accurate predictions for male gender at birth. Additional analysis of further steroids did neither increase the accuracy, sensitivity nor specificity of this prediction. The estimated optimal cut-off value for amniotic testosterone level was 0.074 μg/L for healthy male newborns. LC-MS/MS is a reliable method for the determination of steroids in amniotic fluid. The determination of testosterone in amniotic fluid by LC-MS/MS in early pregnancy of healthy subjects can be used

  2. Neurorescue effects and stem properties of chorionic villi and amniotic progenitor cells.

    Science.gov (United States)

    Calzarossa, C; Bossolasco, P; Besana, A; Manca, M P; De Grada, L; De Coppi, P; Giardino, D; Silani, V; Cova, L

    2013-03-27

    The capability to integrate into degenerative environment, release neurotrophic cytokines, contrast oxidative stress and an inherent differentiation potential towards siteappropriate phenotypes are considered crucial for the use of stem cells in tissue repair and regeneration. Naïve human chorial villi- (hCVCs) and amniotic fluid- (hAFCs) derived cells, whose properties and potentiality have not been extensively investigated, may represent two novel foetal cell sources for stem cell therapy. We previously described that long-term transplantation of hAFCs in the lateral ventricles of wobbler and healthy mice was feasible and safe. In the present study we examine the in vitro intrinsic stem potential of hCVCs and hAFCs for future therapeutic applications in neurodegenerative disorders. Presence of stem lineages was evaluated assessing the expression pattern of relevant candidate markers by flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. Release of cytokines that may potentialy sustain endogenous neurogenesis and/or activate neuroprotective pathways was quantified by enzyme-linked immunosorbent assays (ELISAs). We also performed an in vitro neurorescue assay, wherein a neuroblastoma cell line damaged by 6-hydroxydopamine (6-OHDA) was treated with hCVC/hAFC-derived conditioned medium (CM). Naïve hCVCs/hAFCs show a neurogenic/angiogenic predisposition. Both cell types express several specific neural stem/progenitor markers, such as nestin and connexin 43, and release significant amounts of brain-derived neurotrophic factor, as well as vascular endothelial growth factor. hCVC and hAFC populations comprise several interesting cell lineages, including mesenchymal stem cells (MSCs) and cells with neural-like phenotypes. Moreover, although CMs obtained from both cell cultures actively sustained metabolic activity in a 6-OHDA-induced Parkinson's disease (PD) cell model, only hCVC-derived CMs significantly reduced neurotoxin

  3. Membrana amniótica nas cirurgias reconstrutivas da superfície ocular nas ceratoconjuntivites cicatriciais Amniotic membrane for ocular surface reconstruction in cicatricial keratoconjunctivitis

    Directory of Open Access Journals (Sweden)

    José Alvaro Pereira Gomes

    1999-10-01

    based on the ability to improve epithelial healing and to decrease inflammatory, angiogenic and cicatricial processes. The purpose of this study was to report the surgical outcome of human amniotic membrane use for surface reconstruction in ocular cicatricial diseases. Methods: Amniotic membrane was obtained at the time of cesarean section and was preserved at -80ºC in glycerol and cornea culture media at a ratio of 1:1. Eleven eyes of 10 patients underwent amniotic membrane transplantation, associated (8 eyes or not (3 eyes with corneal limbal graft. Ocular surface reconstruction was performed after chemical burns (6 eyes, trauma (1 eye and Stevens-Johnson's syndrome (SJS (4 eyes. Results: Mean follow-up time was 5.22 months (range, 2-13 months. One case of SSJ developed early postoperative infection and was excluded from the analysis. Successful ocular surface reconstruction was achieved in 8 eyes (80%. Surgical failure was observed in 2 cases of SJS who presented corneal melting at the time of surgery (20%. Conclusions: This study suggests that amniotic membrane transplantation is an effective alternative for surface reconstruction in stable ocular cicatricial diseases. Larger studies with longer follow-up are necessary to further analyse this procedure.

  4. The ontogenetic transformation of the mesosaurid tarsus: a contribution to the origin of the primitive amniotic astragalus

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    Graciela Piñeiro

    2016-05-01

    Full Text Available The hypotheses about the origin of the primitive amniotic tarsus are very speculative. Early studies argued that the origin of the astragalus, one of the largest proximal bones in the tarsus of basal amniotes, was produced by either the fusion of two, three, or even four of the original tarsal bones, the intermedium, the tibiale and the proximal centralia (c4 and c3, or that the intermedium alone transforms into the primitive astragalus. More recent studies have shown that the structure of the tarsus in Captorhinus supports the former hypothesis about a fusion of the intermedium, the tibiale, the proximal centrale (c4 and eventually c3, producing a purportedly multipartite structure of the amniotic astragalus, but the issue remained contentious. Very well preserved tarsi of the Early Permian aquatic amniote Mesosaurus tenuidens Gervais, 1864–1865, which represent the most complete ontogenetic succession known for a basal amniote (the other exceptional one is provided by the Late Permian diapsid Hovasaurus boulei Piveteau, 1926, suggest that there is more than one ossification center for the astragalus and that these fuse during late embryonic stages or maybe early after birth. A non-hatched Mesosaurus in an advanced stage of development shows that the tarsus is represented by a single bone, most probably the astragalus, which seems to be formed by the suturing of three bones, here interpreted as being the intermedium, the tibiale, probably already integrated to the c4 in an earlier stage of the development, and the c3. An amniote-like tarsal structure is observed in very basal Carboniferous and Permian tetrapods such as Proterogyrinus, Gephyrostegus, the diadectids Diadectes and Orobates, some microsaurs like Tuditanus and Pantylus and possibly Westlothiana, taxa that were all considered as true amniotes in their original descriptions. Therefore, the structure of the amniotic tarsus, including the configuration of the proximal series formed by

  5. Isolation, culture, characterization and cryopreservation of stem cells derived from amniotic mesenchymal layer and umbilical cord tissue of bovine fetuses

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    Loreta L. Campos

    Full Text Available ABSTRACT: Stem cells are undifferentiated cells with a high proliferation potential. These cells can be characterized by their in vivo ability to self-renew and to differentiate into specialized cell lines. The most used stem cell types, in both human and veterinary fields, are the mesenchymal stem cells (MSC derived from bone marrow and adipose tissue. Nowadays, there is a great interest in using stem cells derived from fetal tissues, such as amniotic membrane (AM and umbilical cord tissue (UCT, which can be obtained non-invasively at delivery time. Due to the scarcity of studies in bovine species, the aim of this study was to isolate, characterize, differentiate and cryopreserve MSC derived from the mesenchymal layer of amniotic membrane (AM, for the first time, and umbilical cord tissue (UCT of dairy cow neonates after assisted delivery (AD and from fetus at initial third of pregnancy (IT obtained in slaughterhouse. Cells were isolated by enzymatic digestion of the tissue fragments with 0.1% collagenase solution. Six samples of AM and UCT at delivery time and six samples of AM and UCT at first trimester of pregnancy were subjected to morphology evaluation, imunophenotype characterization, in vitro osteogenic, adipogenic and chondrogenic differentiation and viability analysis after cryopreservation. All samples showed adherence to plastic and fibroblast-like morphology. Immunocytochemistry revealed expression of CD 44, NANOG and OCT-4 and lack of expression of MHC II in MSC from all samples. Flow cytometry demonstrated that cells from all samples expressed CD 44, did not or low expressed CD 34 (AM: IT-0.3%a, AD-3.4%b; UCT: 0.4%, 1.4% and MHC II (AM: IT-1.05%a, AD-9.7%b; UCT: IT-0.7%a, AD-5.7%b. They were also capable of trilineage mesenchymal differentiation and showed 80% viability after cryopreservation. According to the results, bovine AM and UCT-derived cells, either obtained at delivery time or from slaughterhouse, are a painless and non

  6. Amniotic fluid protein profiles of intraamniotic inflammatory response to Ureaplasma spp. and other bacteria.

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    Marian Kacerovsky

    Full Text Available OBJECTIVE: This study aimed to evaluate the amniotic fluid protein profiles and the intensity of intraamniotic inflammatory response to Ureaplasma spp. and other bacteria, using the multiplex xMAP technology. METHODS: A retrospective cohort study was undertaken in the Department of Obstetrics and Gynecology, University Hospital Hradec Kralove, Czech Republic. A total of 145 pregnant women with preterm prelabor rupture of membranes between gestational age 24+0 and 36+6 weeks were included in the study. Amniocenteses were performed. The presence of Ureaplasma spp. and other bacteria was evaluated using 16S rRNA gene sequencing. The levels of specific proteins were determined using multiplex xMAP technology. RESULTS: The presence of Ureaplasma spp. and other bacteria in the amniotic fluid was associated with increased levels of interleukin (IL-6, IL-8, IL-10, brain-derived neurotropic factor, granulocyte macrophage colony stimulating factor, monocyte chemotactic protein-1, macrophage inflammatory protein-1, and matrix metalloproteinasis-9. Ureaplasma spp. were also associated with increased levels of neurotropin-3 and triggering receptor expressed on myeloid cells-1. CONCLUSIONS: The presence of Ureaplasma spp. in the amniotic fluid is associated with a slightly different protein profile of inflammatory response, but the intensity of inflammatory response to Ureaplasma spp. is comparable with the inflammatory response to other bacteria.

  7. 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.

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

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    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. The Effect of the Amniotic Fluid Index on the Accuracy of Ultrasonographic-Estimated Fetal Weight.

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    Karahanoglu, Ertugrul; Altinboga, Orhan; Akpinar, Funda; Gultekin, Ismail Burak; Ozdemirci, Safak; Akyol, Aysegul; Yalvac, Serdar

    2017-06-01

    Fetal weight estimation is one of the most important aspects of antenatal care. The effects of amniotic fluid volume on the accuracy of estimated fetal weight (EFW) depend on the amount of fluid, in particular whether it is polyhydramnios or oligohydramnios. Previous studies have reported conflicting results of the effects of amniotic fluid volume on EFW accuracy. The aim of the study was to evaluate the effects of isolated oligohydramnios cases and polyhydramnios on the accuracy of EFW. A retrospective study was conducted at a tertiary center. The study groups consisted of 1069 term isolated oligohydramnios cases, 182 term isolated polyhydramnios cases, and 392 term-matched cases with a normal volume of amniotic fluid. Estimated fetal weight error was determined and expressed in terms of systematic error, calculated from mean percentage error and random error. The systematic error did not differ significantly between polyhydramnios and oligohydramnios cases (-3.60 [8.94%] vs -2.73 [9.7%]). The random error was 8.94% in polyhydramnios cases and 9.7% in oligohydramnios cases. The overestimation rate was 63.6% in polyhydramnios cases and 66.3% in oligohydramnios cases. There were no significant differences in the accuracy of EFW between oligohydramnios and polyhydramnios. However, there was a tendency for overestimation in both types of cases.

  10. Outcome of application of amniotic membrane graft in ocular surface disorders.

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    Ashraf, Nargis Nizam; Adhi, Muhammad Idrees

    2017-07-01

    To determine the outcome of application of amniotic graft in ocular surface disorders. This cross-sectional study was conducted at Dow University of Health Sciences, Karachi, from January 2010 to December 2012, and comprised patients with ocular surface disorders. Patients' presenting symptoms and signs were recorded. Previously harvested and frozen amniotic graft was applied in different types of ocular surface disorders, such as corneal ulcers, pterygium, keratomalacia, Steven-Johnson syndrome, etc. Following the surgery, patients were assessed for improvement in symptoms and signs related to epithelialisation in corneal ulcers.. Of the 50 patients, 30(60%) were male and 20(40%) female. The overall mean age was 40±19.3 years (range: 9 months to 80 years). Out of the 18(36%) cases of pterygium, there was recurrence in 5(27.7%) cases. There were 26(52%) patients of corneal ulcers, of whom re-epithelialisation occurred in 21(80.7%) patients. Amniotic membrane grafting was found to be a safe procedure for ocular surface disorders.

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

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    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.)

  12. Applications of Amniotic Membrane and Fluid in Stem Cell Biology and Regenerative Medicine

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    Kerry Rennie

    2012-01-01

    Full Text Available The amniotic membrane (AM and amniotic fluid (AF have a long history of use in surgical and prenatal diagnostic applications, respectively. In addition, the discovery of cell populations in AM and AF which are widely accessible, nontumorigenic and capable of differentiating into a variety of cell types has stimulated a flurry of research aimed at characterizing the cells and evaluating their potential utility in regenerative medicine. While a major focus of research has been the use of amniotic membrane and fluid in tissue engineering and cell replacement, AM- and AF-derived cells may also have capabilities in protecting and stimulating the repair of injured tissues via paracrine actions, and acting as vectors for biodelivery of exogenous factors to treat injury and diseases. Much progress has been made since the discovery of AM and AF cells with stem cell characteristics nearly a decade ago, but there remain a number of problematic issues stemming from the inherent heterogeneity of these cells as well as inconsistencies in isolation and culturing methods which must be addressed to advance the field towards the development of cell-based therapies. Here, we provide an overview of the recent progress and future perspectives in the use of AM- and AF-derived cells for therapeutic applications.

  13. Bilateral multiple recession coverage with platelet-rich fibrin in comparison with amniotic membrane

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    Sonia S Shetty

    2014-01-01

    Full Text Available Various plastic procedures are done to enhance esthetics, relieve hypersensitivity or even prevent root caries. The most predictable plastic procedure is the coronally advanced flap procedure, with subepithelial connective tissue. Owing to the second surgical donor site and difficulty in procuring a sufficient graft in multiple recessions, various alternative additive membranes are used. This is a case report, the first of its kind, wherein a bilaterally occurring multiple Millers class I recession was managed by using Platelet-rich Fibrin (PrF and amniotic membrane, in a 40-year-old male, who presented to the Department of Periodontics. He complained of hypersensitivity in relation to the upper right and left back region, a bilateral Millers class I recession in relation to 15, 16, and 25, 26 of 3 mm each. Both the recessions were planned for root coverage with coronally advanced flap and additive membrane. The sites were randomly assigned for the use of platelet-rich fibrin and an aminotic membrane. The clinical outcome of the surgical procedure accounted for 100% root coverage, an enhanced gingival biotype, with both the membranes. Furthermore, the results were stable even after seven months in the amniotic membrane-treated site. Hence, the use of amniotic membrane as a novel approach to root coverage is more advantageous than PrF owing to the laboratory preparation of the autologous biomaterial.

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

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    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.

  15. Microbiological viability of bovine amniotic membrane stored in glycerin 99% at room temperature for 48 months

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    Kelly Cristine de Sousa Pontes

    Full Text Available ABSTRACT The medium for storing biological tissues is of great importance for their optimal use in surgery. Glycerin has been proven efficient for storing diverse tissues for prolonged time, but the preservation of the bovine amniotic membrane in glycerin 99% at room temperature has never been evaluated to be used safely in surgical procedures. This study evaluated the preservation of 80 bovine amniotic membrane samples stored in glycerin 99% at room temperature. The samples were randomly divided evenly into four groups. Samples were microbiologically tested after 1, 6, 12 and 48 months of storage. The presence of bacteria and fungi in the samples was evaluated by inoculation on blood agar and incubation at 37 ºC for 48 hours and on Sabouraud agar at 25 ºC for 5 to 10 days. No fungal or bacterial growth was detected in any of the samples. It was concluded that glycerin is an efficient medium, regarding microbiology, for preserving pre-prepared bovine amniotic membrane, keeping the tissue free of microorganisms that grow in the media up to 48 months at room temperature.

  16. Are amniotic fluid alpha-fetoprotein levels influenced by the gender in twin pairs?

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    Sharony, Reuven; Drugan, Arie; Amiel, Aliza; Grinshpun-Cohen, Julia; Markov, Shlomo; Fejgin, Moshe D

    2003-01-01

    To examine the assumption that amniotic fluid alpha-fetoprotein (AFAFP) levels are different in female and male twin fetuses. Amniotic fluid levels of AFP in pregnancies with female and male fetuses in gender-concordant and gender-discordant twins were compared. A t test of p gender (84 females and 82 males) while 166 pairs differed in their gender. The amniotic fluid AFP levels of each sac were measured using fluorescent immunoassay methods by an AutoDELFIA machine. The mean levels of AFAFP were lower in female twins compared to their male counterparts in same-gender twins (p = 0.07), although the difference was quite small. Nevertheless, there was no such difference between AFAFP of male versus female fetuses in gender-discordant twins. The levels of AFAFP were higher in the male twins of gender-concordant twins in comparison to female twins. No such difference was found between female versus male fetuses in gender-disconcordant twins. Copyright 2003 S. Karger AG, Basel

  17. Use of routinely collected amniotic fluid for whole-genome expression analysis of polygenic disorders.

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    Nagy, Gyula Richárd; Gyõrffy, Balázs; Galamb, Orsolya; Molnár, Béla; Nagy, Bálint; Papp, Zoltán

    2006-11-01

    Neural tube defects related to polygenic disorders are the second most common birth defects in the world, but no molecular biologic tests are available to analyze the genes involved in the pathomechanism of these disorders. We explored the use of routinely collected amniotic fluid to characterize the differential gene expression profiles of polygenic disorders. We used oligonucleotide microarrays to analyze amniotic fluid samples obtained from pregnant women carrying fetuses with neural tube defects diagnosed during ultrasound examination. The control samples were obtained from pregnant women who underwent routine genetic amniocentesis because of advanced maternal age (>35 years). We also investigated specific folate-related genes because maternal periconceptional folic acid supplementation has been found to have a protective effect with respect to neural tube defects. Fetal mRNA from amniocytes was successfully isolated, amplified, labeled, and hybridized to whole-genome transcript arrays. We detected differential gene expression profiles between cases and controls. Highlighted genes such as SLA, LST1, and BENE might be important in the development of neural tube defects. None of the specific folate-related genes were in the top 100 associated transcripts. This pilot study demonstrated that a routinely collected amount of amniotic fluid (as small as 6 mL) can provide sufficient RNA to successfully hybridize to expression arrays. Analysis of the differences in fetal gene expressions might help us decipher the complex genetic background of polygenic disorders.

  18. Amniotic fluid phthalate levels and male fetal gonad function

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    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)...

  19. 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

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

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    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.

  1. Amniotic fluid insulin and C-peptide as predictive markers for fetal macrosomia, birth injuries, and delivery complications?

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    Stein, Roland Gregor; Meinusch, Malgorzata; Diessner, Joachim; Dietl, Johannes; Hönig, Arnd; Zollner, Ursula

    2014-01-15

    Gestational diabetes mellitus (GDM) occurs in 3-5% of all pregnancies. GDM increases both maternal and fetal risks, causes fetal macrosomia, and hence increases the rates of caesarean sections and delivery complications such as shoulder dystocia. An early predictive marker and consequent early treatment could be beneficial, so amniotic fluid insulin and C-peptide have been examined in several studies. Increased amniotic fluid insulin in early amniocentesis between the 14th and 20th gestational week predicted a later GDM. A potential direct association with fetal macrosomia remains to be determined. This retrospective study investigated amniotic fluid insulin/C-peptide from amniocenteses between 14 and 20 weeks of gestation in correlation with fetal birth weight, type of delivery, and complications. To focus on effects of fetal hyperinsulinism apart from therapeutic confounders, we included patients who did not participate in GDM screening. Insulin and C-peptide were measured in 144 samples of frozen amniotic fluid. Birth weight, type of delivery, complications, and birth injuries were noted. Birth weights ranged from 760 g to 4410 g with a mean weight of 3424 g at an average of 40 weeks gestation. The mean amniotic fluid insulin was 4.36 U/ml and the mean C-peptide concentration was 0.076 ng/ml. There was no correlation between amniotic fluid insulin or C peptide and birth weight, type of delivery, complications, and birth injuries. Amniotic fluid insulin and C-peptide are unsuitable as predictive marker for fetal macrosomia, type of delivery, complications, or birth injuries.

  2. Impedance-matching hearing in Paleozoic reptiles: evidence of advanced sensory perception at an early stage of amniote evolution.

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    Johannes Müller

    Full Text Available BACKGROUND: Insights into the onset of evolutionary novelties are key to the understanding of amniote origins and diversification. The possession of an impedance-matching tympanic middle ear is characteristic of all terrestrial vertebrates with a sophisticated hearing sense and an adaptively important feature of many modern terrestrial vertebrates. Whereas tympanic ears seem to have evolved multiple times within tetrapods, especially among crown-group members such as frogs, mammals, squamates, turtles, crocodiles, and birds, the presence of true tympanic ears has never been recorded in a Paleozoic amniote, suggesting they evolved fairly recently in amniote history. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we performed a morphological examination and a phylogenetic analysis of poorly known parareptiles from the Middle Permian of the Mezen River Basin in Russia. We recovered a well-supported clade that is characterized by a unique cheek morphology indicative of a tympanum stretching across large parts of the temporal region to an extent not seen in other amniotes, fossil or extant, and a braincase specialized in showing modifications clearly related to an increase in auditory function, unlike the braincase of any other Paleozoic tetrapod. In addition, we estimated the ratio of the tympanum area relative to the stapedial footplate for the basalmost taxon of the clade, which, at 23:1, is in close correspondence to that of modern amniotes capable of efficient impedance-matching hearing. CONCLUSIONS/SIGNIFICANCE: Using modern amniotes as analogues, the possession of an impedance-matching middle ear in these parareptiles suggests unique ecological adaptations potentially related to living in dim-light environments. More importantly, our results demonstrate that already at an early stage of amniote diversification, and prior to the Permo-Triassic extinction event, the complexity of terrestrial vertebrate ecosystems had reached a level that

  3. A point of care test for interleukin-6 in amniotic fluid in preterm prelabor rupture of membranes: a step toward the early treatment of acute intra-amniotic inflammation/infection

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    Chaemsaithong, Piya; Romero, Roberto; Korzeniewski, Steven J.; Martinez-Varea, Alicia; Dong, Zhong; Yoon, Bo Hyun; Hassan, Sonia S.; Chaiworapongsa, Tinnakorn; Yeo, Lami

    2017-01-01

    Objective Preterm prelabor rupture of membranes (preterm PROM) accounts for 30–40% of spontaneous preterm deliveries and thus is a major contributor to perinatal morbidity and mortality. An amniotic fluid (AF) interleukin-6 (IL-6) concentration is a key cytokine for the identification of intra-amniotic inflammation, patients at risk of impending preterm delivery, and adverse pregnancy complications. The conventional method to determine IL-6 concentrations in AF is an enzyme-linked immunosorbent assay (ELISA). However, this technique is not available in clinical settings, and the results may take several days. A lateral flow-based immunoassay, or point of care (POC) test, has been developed to address this issue. The objective of this study was to compare the performance of AF IL-6 determined by the POC test to that determined by ELISA for the identification of intra-amniotic inflammation in patients with preterm PROM. Materials and Methods This retrospective cohort study includes 56 women with singleton pregnancies who presented with preterm PROM. Amniocentesis was performed at the time of diagnosis, and AF was analyzed using cultivation techniques for aerobic and anaerobic bacteria as well as genital mycoplasmas. AF Gram stain and AF white blood cell counts were determined. AF IL-6 concentrations were measured using both lateral flow-based immunoassay and ELISA. The primary outcome was intra-amniotic inflammation defined as AF ELISA IL-6 ≥ 2,600 pg/ml. A previously determined cut-off of 745 pg/ml was used to define a positive POC test. Results 1) The POC test for AF IL-6 concentrations had 97% sensitivity and 96% specificity for the identification of intra-amniotic inflammation, as defined using ELISA among patients with preterm PROM; and 2) the diagnostic performance of the POC test for IL-6 was strongly correlated to that of an ELISA test for the identification of intra-amniotic inflammation and was equivalent for the identification of acute inflammatory

  4. Soluble leukocyte-Associated Ig-like Receptor-1 in amniotic fluid is of fetal origin and positively associates with lung compliance.

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    Michiel L Houben

    Full Text Available The soluble form of the inhibitory immune receptor leukocyte-Associated Ig-like Receptor-1 (sLAIR-1 is present in plasma, urine and synovial fluid and correlates to inflammation. We and others previously showed inflammatory protein expression in normal amniotic fluid at term. We hypothesized that sLAIR-1 is present in amniotic fluid during term parturition and is related to fetal lung function development. sLAIR-1 was detectable in all amniotic fluid samples (n=355 collected during term spontaneous deliveries. First, potential intra-uterine origins of amniotic fluid sLAIR-1 were explored. Although LAIR-1 was expressed on the surface of amniotic fluid neutrophils, LAIR-1 was not secreted upon ex vivo neutrophil stimulation with LPS, or PMA/ionomycin. Cord blood concentrations of sLAIR-1 were fourfold lower than and not related to amniotic fluid concentrations and placentas showed no or only sporadic LAIR-1 positive cells. Similarly, in post-mortem lung tissue of term neonates that died of non-pulmonary disorders LAIR-1 positive cells were absent or only sporadically present. In fetal urine samples, however, sLAIR-1 levels were even higher than in amniotic fluid and correlated with amniotic fluid sLAIR-1 concentrations. Second, the potential relevance of amniotic fluid sLAIR-1 was studied. sLAIR-1 concentrations had low correlation to amniotic fluid cytokines. We measured neonatal lung function in a convenient subset of 152 infants, using the single occlusion technique, at a median age of 34 days (IQR 30-39. The amniotic fluid concentration of sLAIR-1 was independently correlated to airway compliance (ρ=0.29, P=.001. Taken together, we show the consistent presence of sLAIR-1 in amniotic fluid, which originates from fetal urine. Concentrations of sLAIR-1 in amniotic fluid during term deliveries are independent from levels of other soluble immune mediators. The positive association between concentrations of amniotic fluid sLAIR-1 and neonatal lung

  5. 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.

  6. 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.

  7. Expression of a Hoechst 33342 Efflux Phenomenon and Common Characteristics Of Pluripotent Stem Cells in a Side Population of Amniotic Fluid Cells

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    Yieh-Loong Tsai

    2010-06-01

    Conclusion: Amniotic fluid contains a SP that was found mostly among the SAFCs. Enriched SP cells isolated by the efflux of Hoechst 33342 could be a novel and promising source of pluripotent-like amniotic derived stem cells for cellular therapy in the near future.

  8. Multilayer amniotic membrane transplantation for bacterial keratitis with corneal perforation after hyperopic photorefractive keratectomy: case report and literature review.

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    Nubile, Mario; Carpineto, Paolo; Lanzini, Manuela; Ciancaglini, Marco; Zuppardi, Eduardo; Mastropasqua, Leonardo

    2007-09-01

    We report a case of corneal infection caused by Streptococcus pneumoniae after hyperopic photorefractive keratectomy (PRK) that produced severe corneal melting, ulceration, and multiple perforations. Treatment included antibiogram-based antibiotic topical therapy and multilayer amniotic membrane transplantation (AMT) performed to seal the perforations and restore the globe integrity. Clinical and anterior segment optical coherence tomography (Visante OCT, Carl Zeiss Meditec) examinations documented progressive integration of the amniotic membrane tissues within the cornea, stromal and epithelial healing, and recovery of a stable and regular anterior chamber. The cornea healed with an avascular leucoma; the best corrected visual acuity was reduced to 20/200. Severe pneumococcal ulcerative perforation is a potential complication of PRK. Penetrating keratoplasty, at high risk for failure in the acute settings of an infected and inflamed eye, can be delayed until the cornea is healed. Amniotic membrane transplantation may be an alternative surgical option to achieve this goal.

  9. Two rare cases of simultaneous Tessier number 3 cleft, contralateral cleft lip, and signs of amniotic band syndrome.

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    Xu, Yi; Mu, Yue; Chen, Renji; Zheng, Zongmei; Zhang, Wenjing

    2016-06-01

    The Tessier number 3 cleft is rare. In this paper, we report two extremely rare cases of simultaneous Tessier number 3 cleft, contralateral cleft lip, and signs of amniotic band syndrome. In the two cases, we confirmed that amniotic bands were the probable cause of the Tessier number 3 cleft, where swallowed fibrous strands of amniotic bands entangle a typical cleft lip and cause the more severe Tessier number 3 cleft. In this study, Z-plasty was performed for one case, and a straight-line method was used for the other. Postoperatively, the appearance of both patients was satisfactory, as expected. Consequently, treatment for the Tessier number 3 cleft should be designed individually based on the severity of deformity. Copyright © 2016. Published by Elsevier Ltd.

  10. A comparison between amniotic fluid index and the single deepest vertical pocket technique in predicting adverse outcome in prolonged pregnancy

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    Rosati, Paolo; Guariglia, Lorenzo; Cavaliere, Anna Franca; Ciliberti, Paola; Buongiorno, Silvia; Ciardulli, Andrea; Cianci, Stefano; Vitale, Salvatore Giovanni; Cignini, Pietro; Mappa, Ilenia

    2015-01-01

    Objective to compare perinatal outcome in induced postterm pregnancies with normal amniotic volume and in patients with prolonged pregnancy undergone induction for oligohydramnios, evaluated by two different ultrasonographic methods. Methods amniotic fluid volume was measured, using Single Deepest Vertical Pocket (SDVP) and Amniotic Fluid Index (AFI), in 961 singleton uncomplicated prolonged pregnancies. In 109 of these patients, hospitalization was planned for induction of labor, during or after 42 weeks of gestation, for oligohydramnios, postterm pregnancy and other indications in 47, 51 and 11 cases, respectively. Perinatal outcome included: rate of caesarean section, fetal distress, non reassuring fetal heart tracing, presence of meconium, umbilical artery pH postterm versus prolonged pregnancies with oligohydramnios, also in relation to the two different ultrasonographic methods. Conclusions oligohydramnios is more frequently diagnosed using AFI than SDVP, consequently determining a higher rate of induction of labor. Moreover, perinatal outcome in prolonged induced pregnancies is not affected by oligohydramnios. PMID:26918093

  11. Successful treatment of atrial flutter by repeated intraperitoneal and intra-amniotic injections of amiodarone in a fetus with hydrops.

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    Lin, Pei-Hsuan; Wu, Hsin-Hung; Tsai, Horng-Der; Hsieh, Charles Tsung-Che

    2016-06-01

    We report a case of nonimmune hydrops fetalis caused by atrial flutter, which was successfully treated by intraperitoneal and intra-amniotic injections of amiodarone. A 27-year-old woman presented at 30 weeks of pregnancy with hydrops fetalis caused by a fetal atrial flutter. As the transplacental passage of antiarrhythmic agents is impaired in hydrops fetalis, we chose direct treatment using fetal intraperitoneal and intra-amniotic injections (75-300 mg) of amiodarone. We managed to successfully convert the fetal atrial flutter to normal sinus rhythm. The woman delivered a live female baby at 33 weeks of gestation with normal sinus rhythm and neurological development. Intrauterine antiarrhythmic treatment can reduce perinatal morbidity and mortality. This report suggests that direct fetal therapy using intraperitoneal or intra-amniotic injections of amiodarone constitutes an effective treatment for atrial flutter in cases of hydrops fetalis. Copyright © 2016. Published by Elsevier B.V.

  12. Determinants of organochlorine levels detectable in the amniotic fluid of women from Tenerife Island (Canary Islands, Spain).

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    Luzardo, Octavio P; Mahtani, Vikesh; Troyano, Juan M; Alvarez de la Rosa, Margarita; Padilla-Pérez, Ana I; Zumbado, Manuel; Almeida, Maira; Burillo-Putze, Guillermo; Boada, Carlos; Boada, Luis D

    2009-07-01

    Organochlorines (OCs) tend to accumulate in human tissues and can be measured in amniotic fluid (AF). The detection of OCs in AF samples reflects intrauterine exposure of human beings to these persistent organic pollutants. The present study was performed to evaluate the level of contamination of AF by OCs in 100 pregnant women from Tenerife Island (Canary Islands, Spain). Gas chromatography/mass spectrometry (GC/MS) was used to identify and quantify the analytes, including 7 polychlorobiphenyl (PCB) congeners and 18 OC pesticides and metabolites. The majority of the AF samples (67%) showed some detectable OC-residue, hexachlorobenzene (HCB) being the most frequently detected compound (66% of the samples) and at the highest concentration (median 0.023 ng/ml). Lindane was also detected in 28% of the samples. Inverse associations were found between previous lactation and hexachlorocyclohexane isomers (HCH) and cyclodienes in the group of younger women (p = 0.037 and p = 0.027, respectively). Unexpectedly, serum values of HCB (r = -0.414; p = 0.04), gamma-HCH (r = -0.294; p = 0.035), and SigmaOCs (r = -0.350; p = 0.014) were negatively related to age. Even more, women with detectable levels of HCH isomers were younger (33.9 +/- 4.9 years) than women with undetectable levels of them (36.1 +/- 4.9 years; p = 0.035). We conclude that approximately one in two fetuses in the Canary Islands is exposed to OCs in utero, and that, therefore, the exposure of young women from these Islands to some HCH isomers persists nowadays. Because prenatal exposure to these chemicals may be a causative factor in adverse health trends, further studies are required to enhance preventive measures.

  13. Application of fresh amniotic membrane transplantation combined therapeutic soft corneal contact lens in pterygium

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    Han-Yuan Ye

    2014-09-01

    Full Text Available AIM:To evaluate the clinical effect of therapeutic soft corneal contact lens in combination with fresh amniotic membrane transplantation in the treatment of pterygium and its clinical value.METHODS:This study was a retrospective analysis of 200 pterygium excision patients(200 cases, 220 eyesin hospital from January 2010-June 2011. The patients were divided into two groups. Group 1(100 cases, 110 eyeswere patients treated with fresh amniotic membrane transplantation and therapeutic soft corneal contact lens while group 2(control group, 100 cases, 110 eyesdid not use corneal contact lens. The clinical outcomes in these two groups were compared in a 6-18mo postoperative follow-up observation.RESULTS:In control group using corneal contact lens the epithelium repair time ranged from 2-7d with an average time of 3.8d; in group 1 the epithelial repair time ranged from 1-5d with an average time of 2.5d. The patients were followed up for 6-18mo, 9 recurrent eyes were found in control group, the recurrence rate was 8.2%; 5 recurrent eyes were found in experimental group the recurreat rate was 4.5%. There were significant differences in the 2 groups comparing treatment outcomes(PCONCLUSION:The therapeutic effect of fresh amniotic membrane transplantation after pterygium excision is enhanced by the therapeutic application of corneal contact lens. The epithelial repair time is shortened, the recurrence rate is reduced after pterygium excision and stimulative symptoms including photophobia, tearing, foreign body sensation and ophthalmodynia caused by corneal epithelial defect can be relieved.

  14. Recurrent cubital tunnel syndrome treated with revision neurolysis and amniotic membrane nerve wrapping.

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    Gaspar, Michael P; Abdelfattah, Hesham M; Welch, Ian W; Vosbikian, Michael M; Kane, Patrick M; Rekant, Mark S

    2016-12-01

    Perineural scarring of the ulnar nerve is a predominant cause of symptom recurrence after surgical treatment for primary cubital tunnel syndrome (CuTS). We report our preliminary experience in revision ulnar nerve decompression and nerve wrapping with an amniotic membrane allograft adhesion barrier for treatment of recurrent CuTS. We performed a retrospective review with prospective follow-up of patients with recurrent CuTS who were treated with revision neurolysis with amniotic membrane nerve wrapping. Preoperative elbow motion, grip and pinch strengths, pain level on the visual analog scale level, and the 11-item version of the Disabilities of the Arm, Shoulder and Hand functional outcome score were compared with postoperative values using paired t testing. Symptom characteristics, physical examination findings, complications, and level of satisfaction were also obtained. Eight patients (mean age, 47.5 years) who had undergone at least 2 prior ulnar nerve operations satisfied study inclusion. At mean postoperative follow-up of 30 months, significant improvements were noted across all patients in visual analog scale pain levels (-3.5 vs. preoperatively; P < .0001), 11-item version of the Disabilities of the Arm, Shoulder and Hand scores (-30 vs. preoperatively; P < .0001), and grip strength (+25 pounds vs. preoperatively; P < .0001). Pinch strength and elbow motion were also significantly improved for those patients with comparative preoperative data available. All patients expressed subjective satisfaction with their results. No adverse reactions or complications occurred in any patients. Ulnar nerve wrapping with amniotic membrane allograft, when combined with revision neurolysis, was a safe and subjectively effective treatment for patients with debilitating recurrent CuTS. Copyright © 2016 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.

  15. Intraoperative mitomycin C and amniotic membrane transplantation for fornix reconstruction in severe cicatricial ocular surface diseases.

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    Tseng, Scheffer C G; Di Pascuale, Mario A; Liu, Daniel Tzong-Shyue; Gao, Ying Ying; Baradaran-Rafii, Alireza

    2005-05-01

    To investigate whether intraoperative application of mitomycin C may enhance the success of amniotic membrane transplantation in symblepharon lysis and fornix reconstruction in severe cicatricial ocular surface diseases. Noncomparative interventional case series. Sixteen patients (8 female, 8 male; 18 eyes) with a mean age of 41+/-23.4 years (range, 3-79) and suffering from severe chemical/thermal burns (7 eyes), multiple recurrent pterygia and pseudopterygia (5 eyes), Stevens-Johnson syndrome (4 eyes), and ocular cicatricial pemphigoid (2 eyes) were consecutively enrolled. All except for 2 eyes had had prior surgical attempts of surgical reconstruction, including 6 eyes with a mucous membrane graft (MMG), but still presented with symblepharon and persistent ocular surface inflammation. After excision of subconjunctival fibrovascular tissues, 0.04% mitomycin C was applied for 5 minutes in the deep fornix before amniotic membrane transplantation. Deeper fornix, noninflamed ocular surface, and full motility. The mean epithelial healing time was 4.2+/-1.9 weeks. During the follow-up of 14.16+/-5.2 months, all eyes showed a marked reduction of conjunctival inflammation, a deep fornix, and a continuous tear meniscus. Of 12 eyes with motility restriction, 2 eyes with multiple recurrent pterygia and 1 eye with severe thermal burn showed recurrence of partial motility restriction 2 months after surgery. The vision of 9 eyes was successfully restored by an additional keratolimbal allograft with subsequent penetrating keratoplasty (6 eyes). Intraoperative application of mitomycin C is an effective means to reduce chronic and deep-seated conjunctival inflammation, and helps amniotic membrane restore a deep fornix after symblepharon lysis, even in eyes that had a failed MMG. Restoration of deep fornix and tear meniscus is an important prerequisite to achieve successful reconstruction by subsequent limbal stem cell transplantation.

  16. Identification of a novel Gig2 gene family specific to non-amniote vertebrates.

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    Yi-Bing Zhang

    Full Text Available Gig2 (grass carp reovirus (GCRV-induced gene 2 is first identified as a novel fish interferon (IFN-stimulated gene (ISG. Overexpression of a zebrafish Gig2 gene can protect cultured fish cells from virus infection. In the present study, we identify a novel gene family that is comprised of genes homologous to the previously characterized Gig2. EST/GSS search and in silico cloning identify 190 Gig2 homologous genes in 51 vertebrate species ranged from lampreys to amphibians. Further large-scale search of vertebrate and invertebrate genome databases indicate that Gig2 gene family is specific to non-amniotes including lampreys, sharks/rays, ray-finned fishes and amphibians. Phylogenetic analysis and synteny analysis reveal lineage-specific expansion of Gig2 gene family and also provide valuable evidence for the fish-specific genome duplication (FSGD hypothesis. Although Gig2 family proteins exhibit no significant sequence similarity to any known proteins, a typical Gig2 protein appears to consist of two conserved parts: an N-terminus that bears very low homology to the catalytic domains of poly(ADP-ribose polymerases (PARPs, and a novel C-terminal domain that is unique to this gene family. Expression profiling of zebrafish Gig2 family genes shows that some duplicate pairs have diverged in function via acquisition of novel spatial and/or temporal expression under stresses. The specificity of this gene family to non-amniotes might contribute to a large extent to distinct physiology in non-amniote vertebrates.

  17. Life History Traits, Protein Evolution, and the Nearly Neutral Theory in Amniotes.

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    Figuet, Emeric; Nabholz, Benoît; Bonneau, Manon; Mas Carrio, Eduard; Nadachowska-Brzyska, Krystyna; Ellegren, Hans; Galtier, Nicolas

    2016-06-01

    The nearly neutral theory of molecular evolution predicts that small populations should accumulate deleterious mutations at a faster rate than large populations. The analysis of nonsynonymous (dN) versus synonymous (dS) substitution rates in birds versus mammals, however, has provided contradictory results, questioning the generality of the nearly neutral theory. Here we analyzed the impact of life history traits, taken as proxies of the effective population size, on molecular evolutionary and population genetic processes in amniotes, including the so far neglected reptiles. We report a strong effect of species body mass, longevity, and age of sexual maturity on genome-wide patterns of polymorphism and divergence across the major groups of amniotes, in agreement with the nearly neutral theory. Our results indicate that the rate of protein evolution in amniotes is determined in the first place by the efficiency of purifying selection against deleterious mutations-and this is true of both radical and conservative amino acid changes. Interestingly, the among-species distribution of dN/dS in birds did not follow this general trend: dN/dS was not higher in large, long-lived than in small, short-lived species of birds. We show that this unexpected pattern is not due to a more narrow range of life history traits, a lack of correlation between traits and Ne, or a peculiar distribution of fitness effects of mutations in birds. Our analysis therefore highlights the bird dN/dS ratio as a molecular evolutionary paradox and a challenge for future research. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Interaction between pulmonary surfactant and vernix: a potential mechanism for induction of amniotic fluid turbidity.

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    Narendran, V; Wickett, R R; Pickens, W L; Hoath, S B

    2000-07-01

    The development of amniotic fluid turbidity during the third trimester is a known marker of fetal lung maturity. We hypothesized that this turbidity results from detachment of vernix caseosa from the fetal skin secondary to interaction with pulmonary-derived phospholipids in the amniotic fluid. To test this hypothesis, we exposed vernix to bovine-derived pulmonary surfactant over a physiologically relevant concentration range. Ten milligrams of vernix was evenly applied to the interior walls of 1.5-mL polypropylene microfuge tubes. Surfactant phospholipids were added to the tubes followed by slow rotation at 37 degrees C overnight. The liquid was decanted and spectrophotometrically analyzed at 650 nm to detect solution turbidity due to vernix detachment and/or emulsification. Increasing concentrations of surfactant phospholipids produced a dose-dependent increase in solution turbidity. A phospholipid mixture closely approximating natural pulmonary surfactant but devoid of surfactant-associated proteins yielded no increase. In other studies, the flow properties of vernix were studied in a Haake flow rheometer at 23 degrees C and 37 degrees C. There was a marked temperature-dependent effect with lower stress required to elicit flow at 37 degrees C compared with 23 degrees C. This temperature dependence was also demonstrated in the turbidity assay with a 124% increase in turbidity at body temperature compared with room temperature. We conclude that under in vitro conditions, pulmonary surfactant interacts with vernix resulting in detachment from a solid phase support. We speculate that in utero, this phenomenon contributes to the increase in amniotic fluid turbidity that is observed near term.

  19. ROLE OF ECHOGENIC AMNIOTIC FLUID PARTICLES AND OPTICAL DENSITY IN PREDICTION OF RESPIRATORY DISTRESS SYNDROME AND LABOR

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    Dr. Shankar H S Ram

    2010-01-01

    Full Text Available 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 antenatal visits. The criteria for AFPS score which are taken into account are the amniotic fluid particle size, number and distribution. Serial AFPS measurements were done till onset of labor. AFPS was correlated to AFOD value at spontaneous labor in 123 women. Uncentrifuged fresh amniotic fluid samples were obtained during ARM/amniotomy and used for AFOD estimation at 650 nm. The mean AFPS and AFOD at onset of labor was found to be 5.14 ± 0.69 mm (3.67 – 6.7 CI 95% and 1.03 ± 0.31 (0.35 -1.69 CI 95% respectively in 116 women who delivered normal babies devoid of respiratory distress syndrome (RDS. Serial AFPS measurements showed a definite AFOD surge after a value in the region of 3.8 mm which is obtained culminating in onset of Labor. 28 women (24.1% had dense clusters of free floating particles across the vertical pool in amniotic fluid with mean AFPS and AFOD of 5.6 ± 0.68 mm and 1.12 ± 0.21 respectively. In 123 women, AFPS < 3.8 mm had sensitivity of 85.74% and positive predictive value of 66.67% in predicting RDS. AFPS serves as a sonological marker for fetal lung maturity and labor. The range of AFOD values can be measured in terms of AFPS (r =0.6, F = 69.8, β= + 0.23, p < 0.001. Serial AFPS estimation predicts fetal maturity and onset of labor.

  20. Comparison of amniotic membrane transplantation for all the cornea and bandage contact lens in the treatment for large pterygium surgery

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    Ying-Wei Wang

    2018-01-01

    Full Text Available AIM: To investigate the amniotic membrane transplantation for all the cornea or bandage contact lens after pterygium excision combine with limbal stem cell transplantation in the treatment of large pterygium. METHODS:The 40 eyes of larger pterygium was randomly divided into 2 groups, one group, 20 eyes, with autologous limbal stem cell transplantation combine with amniotic membrane transplantation for all the cornea(amniotic membrane group; another group, 20 eyes, with bandage contact lens after limbal stem cell transplantation(bandage contact lens group. The surgery time, eye comfort postoperation, progress in corneal healing were observed at 1, 3d, 1 and 3wk. RESULTS:The average surgery time of amniotic membrane transplantation group was 61.4±5.2min, and the bandage contact lens group was 34.5±2.7min, which was significantly shorter(Student's t-test, PPP>0.05. The corneal healing score of the two groups was 0.85±0.18 and 1.15±0.18 in the 3wk after operation respectively, and the difference between the two groups was not statistically significant(Student's t-test, P=0.25. There was only one case of recurrence in amniotic membrane group. CONCLUSION:Treatment of larger pterygium with amniotic membrane transplantation for all the cornea or wearing bandage contact lenses after autologous limbal stem cell transplantation, can both ease eye symptoms and promote corneal wound healing. Those patients with bandage contact lens feel more comfortable and convenient than with autologous limbal stem cell transplantation. Also the bandage contact lens surgery is more economical than the other one.

  1. 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.

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    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.

  2. Group B streptococcal infection of the choriodecidua induces dysfunction of the cytokeratin network in amniotic epithelium: a pathway to membrane weakening.

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    Jeroen P Vanderhoeven

    2014-03-01

    Full Text Available Early events leading to intrauterine infection remain poorly defined, but may hold the key to preventing preterm delivery. To determine molecular pathways within fetal membranes (chorioamnion associated with early choriodecidual infection that may progress to preterm premature rupture of membranes (PPROM, we examined the effects of a Group B Streptococcus (GBS choriodecidual infection on chorioamnion in a nonhuman primate model. Ten chronically catheterized pregnant monkeys (Macaca nemestrina at 118-125 days gestation (term = 172 days received choriodecidual inoculation of either GBS (n = 5 or saline (n = 5. Cesarean section was performed in the first week after GBS or saline inoculation. RNA extracted from chorioamnion (inoculation site was profiled by microarray. Single gene, Gene Set, and Ingenuity Pathway Analysis results were validated using qRT-PCR (chorioamnion, Luminex (amniotic fluid, AF, immunohistochemistry, and transmission electron microscopy (TEM. Despite uterine quiescence in most cases, significant elevations of AF cytokines (TNF-α, IL-8, IL-1β, IL-6 were detected in GBS versus controls (p2-fold change, p<0.05. Remarkably, GBS exposure was associated with significantly downregulated expression of multiple cytokeratin (CK and other cytoskeletal genes critical for maintenance of tissue tensile strength. Immunofluorescence revealed highly significant changes in the CK network within amniocytes with dense CK aggregates and retraction from the cell periphery (all p = 0.006. In human pregnancies affected by PPROM, there was further evidence of CK network retraction with significantly shorter amniocyte foot processes (p = 0.002. These results suggest early choriodecidual infection results in decreased cellular membrane integrity and tensile strength via dysfunction of CK networks. Downregulation of CK expression and perturbations in the amniotic epithelial cell intermediate filament network occur after GBS

  3. Effects of intra-amniotic lipopolysaccharide and maternal betamethasone on brain inflammation in fetal sheep.

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    Elke Kuypers

    Full Text Available Chorioamnionitis and antenatal glucocorticoids are common exposures for preterm infants and can affect the fetal brain, contributing to cognitive and motor deficits in preterm infants. The effects of antenatal glucocorticoids on the brain in the setting of chorioamnionitis are unknown. We hypothesized that antenatal glucocorticoids would modulate inflammation in the brain and prevent hippocampal and white matter injury after intra-amniotic lipopolysaccharide (LPS exposure.Time-mated ewes received saline (control, an intra-amniotic injection of 10 mg LPS at 106d GA or 113d GA, maternal intra-muscular betamethasone (0.5 mg/kg maternal weight alone at 113d GA, betamethasone at 106d GA before LPS or betamethasone at 113d GA after LPS. Animals were delivered at 120d GA (term=150d. Brain structure volumes were measured on T2-weighted MRI images. The subcortical white matter (SCWM, periventricular white matter (PVWM and hippocampus were analyzed for microglia, astrocytes, apoptosis, proliferation, myelin and pre-synaptic vesicles.LPS and/or betamethasone exposure at different time-points during gestation did not alter brain structure volumes on MRI. Betamethasone alone did not alter any of the measurements. Intra-amniotic LPS at 106d or 113d GA induced inflammation as indicated by increased microglial and astrocyte recruitment which was paralleled by increased apoptosis and hypomyelination in the SCWM and decreased synaptophysin density in the hippocampus. Betamethasone before the LPS exposure at 113d GA prevented microglial activation and the decrease in synaptophysin. Betamethasone after LPS exposure increased microglial infiltration and apoptosis.Intra-uterine LPS exposure for 7d or 14d before delivery induced inflammation and injury in the fetal white matter and hippocampus. Antenatal glucocorticoids aggravated the inflammatory changes in the brain caused by pre-existing intra-amniotic inflammation. Antenatal glucocorticoids prior to LPS reduced

  4. Amniotic membrane transplantation with topical interferon alfa-2b after excision of ocular surface squamous neoplasia

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    Hua-Tao Xie

    2018-01-01

    Full Text Available To evaluate the outcome of amniotic membrane transplantation (AMT after tumor excision followed by topical interferon alfa-2b (IFNα2b drops for primary ocular surface squamous neoplasia (OSSN. Twelve eyes of 12 patients with a mean age of 66±10y were included. The average follow-up was 23±10mo. All 12 patients had limbal involvement. Smooth ocular surface and transparent cornea were achieved in all cases. No sign of inflammation, neovascularization, symblepharon or recurrence was noted at the last follow-up. We conclude that AMT with topical IFNα2b drops restores a healthy ocular surface in OSSN without recurrence.

  5. Amniotic fluid turbidity: a useful adjunct for assessing fetal pulmonary maturity status.

    Science.gov (United States)

    Strong, T H; Hayes, A S; Sawyer, A T; Folkestad, B; Mills, S; Sugden, P

    1992-06-01

    A rapid, very simple technique for establishing fetal pulmonary maturity status is presented. Among 100 receiving amniocenteses, aspiration of turbid amniotic fluid that would not permit the reading of newsprint through it was associated with a lecithin/sphingomyelin (LS) ratio of greater than or equal to 2.0, or the presence of phosphatidyl glycerol (PG) in 97% (specificity 98%, positive predictive value 97%). The authors conclude that when turbid fluid is aspirated, delay until LS and PG results are known may not be necessary.

  6. Yolk sac development in lizards (Lacertilia: Scincidae): New perspectives on the egg of amniotes.

    Science.gov (United States)

    Stewart, James R; Thompson, Michael B

    2017-04-01

    Embryos of oviparous reptiles develop on the surface of a large mass of yolk, which they metabolize to become relatively large hatchlings. Access to the yolk is provided by tissues growing outward from the embryo to cover the surface of the yolk. A key feature of yolk sac development is a dedicated blood vascular system to communicate with the embryo. The best known model for yolk sac development and function of oviparous amniotes is based on numerous studies of birds, primarily domestic chickens. In this model, the vascular yolk sac forms the perimeter of the large yolk mass and is lined by a specialized epithelium, which takes up, processes and transports yolk nutrients to the yolk sac blood vessels. Studies of lizard yolk sac development, dating to more than 100 years ago, report characteristics inconsistent with this model. We compared development of the yolk sac from oviposition to near hatching in embryonic series of three species of oviparous scincid lizards to consider congruence with the pattern described for birds. Our findings reinforce results of prior studies indicating that squamate reptiles mobilize and metabolize the large yolk reserves in their eggs through a process unknown in other amniotes. Development of the yolk sac of lizards differs from birds in four primary characteristics, migration of mesoderm, proliferation of endoderm, vascular development and cellular diversity within the yolk sac cavity. Notably, all of the yolk is incorporated into cells relatively early in development and endodermal cells within the yolk sac cavity align along blood vessels which course throughout the yolk sac cavity. The pattern of uptake of yolk by endodermal cells indicates that the mechanism of yolk metabolism differs between lizards and birds and that the evolution of a fundamental characteristic of embryonic nutrition diverged in these two lineages. Attributes of the yolk sac of squamates reveal the existence of phylogenetic diversity among amniote lineages

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

    Science.gov (United States)

    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

  8. 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

    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...... and colostrum, and we describe how these fluids may have a therapeutic potential for GI conditions in some pediatric patients, particularly preterm infants. The composition of the two fluids varies widely among different species and the effects are likely highly species specific. Some effects may however...

  9. First report of distal obstructive uropathy and prune-belly syndrome in an infant with amniotic band syndrome.

    Science.gov (United States)

    Chen, C P; Liu, F F; Jan, S W; Wang, K G; Lan, C C

    1997-01-01

    We describe the first report of distal obstructive uropathy and prune-belly syndrome in an infant with amniotic band syndrome. Prenatal ultrasonographic examination in the third trimester revealed intermittent oligohydramnios, bilateral hydronephrosis, and megacystis. Postnatally, the infant was found to have a scalp defect, a skin pedicle, pseudosyndactyly and constriction rings on the hands, marked distention of the abdomen, a fibrous band attached to the proximal urethra causing urethral stricture, a swollen penile shaft, bilateral talipes equinovarus, and syndactyly of the feet. Multiple fibrous amniotic bands could be identified in the placenta. Our case shows that fetal distal obstructive uropathy can be associated with the congenital constriction band syndrome.

  10. A rapid interleukin-6 bedside test for the identification of intra-amniotic inflammation in preterm labor with intact membranes

    Science.gov (United States)

    Chaemsaithong, Piya; Romero, Roberto; Korzeniewski, Steven J.; Martinez-Varea, Alicia; Dong, Zhong; Yoon, Bo Hyun; Hassan, Sonia S.; Chaiworapongsa, Tinnakorn; Yeo, Lami

    2016-01-01

    Abstract Objective: Preterm birth is associated with 5–18% of pregnancies and is the leading cause of neonatal morbidity and mortality. Amniotic fluid (AF) interleukin-6 (IL-6) is a key cytokine for the identification of intra-amniotic inflammation, and patients with an elevated AF IL-6 are at risk for impending preterm delivery. However, results of the conventional method of measurement (enzyme-linked immunosorbent assay; ELISA) are usually not available in time to inform care. The objective of this study was to determine whether a point of care (POC) test or lateral-flow-based immunoassay for measurement of AF IL-6 concentrations can identify patients with intra-amniotic inflammation and/or infection and those destined to deliver spontaneously before term among women with preterm labor and intact membranes. Methods: One-hundred thirty-six women with singleton pregnancies who presented with symptoms of preterm labor and underwent amniocentesis were included in this study. Amniocentesis was performed at the time of diagnosis of preterm labor. AF Gram stain and AF white blood cell counts were determined. Microbial invasion of the amniotic cavity (MIAC) was defined according to the results of AF culture (aerobic and anaerobic as well as genital mycoplasmas). AF IL-6 concentrations were determined by both lateral flow-based immunoassay and ELISA. The primary outcome was intra-amniotic inflammation, defined as AF ELISA IL-6 ≥ 2600 pg/ml. Results: (1) AF IL-6 concentrations determined by a POC test have high sensitivity (93%), specificity (91%) and a positive likelihood ratio of 10 for the identification of intra-amniotic inflammation by using a threshold of 745 pg/ml; (2) the POC test and ELISA for IL-6 perform similarly in the identification of MIAC, acute inflammatory lesions of placenta and patients at risk of impending spontaneous preterm delivery. Conclusion: A POC AF IL-6 test can identify intra-amniotic inflammation in women who present with preterm

  11. Amniotic fluid oxidant-antioxidant status in foetal congenital nervous system anomalies.

    Science.gov (United States)

    Cim, Numan; Tolunay, Harun Egemen; Karaman, Erbil; Boza, Barıs; Bilici, Mustafa; Çetin, Orkun; Yıldızhan, Recep; Sahin, Hanım Guler

    2017-01-01

    Objective This study aimed to evaluate the oxidant-antioxidant status of amniotic fluid in pregnant women with foetal congenital malformations of the central nervous system. Methods We studied pregnant women with foetal congenital nervous system anomalies at 16-22 weeks' gestation (n = 36). The control group (n = 30) consisted of pregnant women at the same gestational age who underwent amniocentesis, resulting in a normal karyotype. We analysed glutathione, catalase, and malondialdehyde levels in amniotic fluid. Enzyme activation was measured by spectrophotometry. Results The demographic features of the groups were similar in terms of age, parity, body mass index, and gestational weeks. We detected lower glutathione and catalase levels in the foetal congenital anomaly group than in the control group. We detected higher malondialdehyde levels in the foetal congenital anomaly group than in the control group. Conclusion In the organism, the rate of formation of free radicals and their rate of removal are balanced, and this is called oxidative balance. As long as oxidative stability is achieved, the organism is not affected by free radicals. This fact should be kept in mind to avoid any type of teratogenic agent that could lead to congenital disorders.

  12. Biochemical profile of amniotic fluid for the assessment of fetal and renal development

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    F.R. Oliveira

    2002-02-01

    Full Text Available Creatinine plays a key role in the function and maturation of fetal kidneys throughout pregnancy. It is important to identify other markers that may help in the diagnosis of renal dysfunction. Our aim was to determine the profile of and the correlation between biochemical markers to be used to assess renal function and maturation of the fetus in the amniotic fluid during pregnancy and to determine the distribution of normal values for creatinine, N-acetyl-ß-D-glucosaminidase (NAG, ß2-microglobulin, glucose, urea, sodium, potassium, phosphorus, calcium, uric acid, albumin, and osmolality in three gestational age groups. This was a cross-section study that assessed 115 samples of amniotic fluid during three different periods of pregnancy, i.e., 13 to 20, 27 to 34, and 36 to 42 weeks. Concentrations of creatinine, NAG, urea, potassium and uric acid increased during pregnancy (P0.6, P0.5, P<0.05. NAG, sodium, albumin and osmolality did not show significant correlations (r<0.5, P<0.05. These tests confirmed the important role of creatinine in terms of correlation with gestational age. ß2-Microglobulin, glucose and uric acid were significant as markers of function and maturation of fetal kidneys, whereas NAG did not demonstrate a useful role for the assessment of renal maturation.

  13. Early fish myoseptal cells: insights from the trout and relationships with amniote axial tenocytes.

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    Yoann Bricard

    Full Text Available The trunk muscle in fish is organized as longitudinal series of myomeres which are separated by sheets of connective tissue called myoseptum to which myofibers attach. In this study we show in the trout that the myoseptum separating two somites is initially acellular and composed of matricial components such as fibronectin, laminin and collagen I. However, myoseptal cells forming a continuum with skeletogenic cells surrounding axial structures are observed between adjacent myotomes after the completion of somitogenesis. The myoseptal cells do not express myogenic markers such as Pax3, Pax7 and myogenin but express several tendon-associated collagens including col1a1, col5a2 and col12a1 and angiopoietin-like 7, which is a secreted molecule involved in matrix remodelling. Using col1a1 as a marker gene, we observed in developing trout embryo an initial labelling in disseminating cells ventral to the myotome. Later, labelled cells were found more dorsally encircling the notochord or invading the intermyotomal space. This opens the possibility that the sclerotome gives rise not only to skeletogenic mesenchymal cells, as previously reported, but also to myoseptal cells. We furthermore show that myoseptal cells differ from skeletogenic cells found around the notochord by the specific expression of Scleraxis, a distinctive marker of tendon cells in amniotes. In conclusion, the location, the molecular signature and the possible sclerotomal origin of the myoseptal cells suggest that the fish myoseptal cells are homologous to the axial tenocytes in amniotes.

  14. [Outcome of conjunctival autograft with amniotic membrane transplantation for recurrent pterygium].

    Science.gov (United States)

    Tanaka, Atsuko; Chikama, Tai-ichiro; Harada, Daisuke; Kawamoto, Koji; Yamada, Naoyuki; Morishige, Naoyuki; Nishida, Teruo

    2011-04-01

    To investigate the outcome of conjunctival autograft with amniotic membrane transplantation without the use of mitomycin C for cases of recurrent pterygium. Thirty-nine eyes of 35 patients (aged 27 to 76 years) who underwent the surgical procedure for recurrent pterygium at Yamaguchi University Hospital from November 1998 to July 2007 were evaluated retrospectively from their medical records. Patients who did not undergo postoperative follow-up examination for at least 6 months were excluded. The mean number of prior surgeries was 2.9 (range, 2 to 10), and the mean +/- SD follow-up time was 42.0 +/- 29.6 months. Twenty seven of 29 eyes (93.1%) with recurrent grade 3 pterygium showed a reduction in the size of the new growth after surgery. The symblepharon improved postoperatively in 5 of 15 eyes (33.3%) and restriction of ocular movement improved in 11 of 24 eyes (45.8%). Most recurrences occurred within 6 months after surgery, although one case did not show a recurrence until 2 years postsurgery. Conjunctival autograft with amniotic membrane transplantation without the use of mitomycin C reduced the size of the recurrent pterygium. However, 6 of the 16 eyes (37.5%) required subsequent surgery, suggesting that the procedure has limitations. Mitomycin C should therefore possibly be considered for cases with severe symblepharon or restriction of ocular movement. In choosing the surgical procedure for recurrent pterygium, diplopia can be an important indicator of the severity of a symblepharon or restriction of ocular movement.

  15. Evolution of the hypoxia-sensitive cells involved in amniote respiratory reflexes.

    Science.gov (United States)

    Hockman, Dorit; Burns, Alan J; Schlosser, Gerhard; Gates, Keith P; Jevans, Benjamin; Mongera, Alessandro; Fisher, Shannon; Unlu, Gokhan; Knapik, Ela W; Kaufman, Charles K; Mosimann, Christian; Zon, Leonard I; Lancman, Joseph J; Dong, P Duc S; Lickert, Heiko; Tucker, Abigail S; Baker, Clare V H

    2017-04-07

    The evolutionary origins of the hypoxia-sensitive cells that trigger amniote respiratory reflexes - carotid body glomus cells, and 'pulmonary neuroendocrine cells' (PNECs) - are obscure. Homology has been proposed between glomus cells, which are neural crest-derived, and the hypoxia-sensitive 'neuroepithelial cells' (NECs) of fish gills, whose embryonic origin is unknown. NECs have also been likened to PNECs, which differentiate in situ within lung airway epithelia. Using genetic lineage-tracing and neural crest-deficient mutants in zebrafish, and physical fate-mapping in frog and lamprey, we find that NECs are not neural crest-derived, but endoderm-derived, like PNECs, whose endodermal origin we confirm. We discover neural crest-derived catecholaminergic cells associated with zebrafish pharyngeal arch blood vessels, and propose a new model for amniote hypoxia-sensitive cell evolution: endoderm-derived NECs were retained as PNECs, while the carotid body evolved via the aggregation of neural crest-derived catecholaminergic (chromaffin) cells already associated with blood vessels in anamniote pharyngeal arches.

  16. Novel implantable composite biomaterial by fibrin glue and amniotic membrane for ocular surface reconstruction.

    Science.gov (United States)

    Cai, Mingming; Zhang, Jie; Guan, Lili; Zhao, Min

    2015-03-01

    Amniotic membrane transplantation (AMT) is considered a substantial treatment option in the management of ocular surface disorders. However, several inherent drawbacks still remain. The present study devised a novel implantable composite biomaterial of fibrin glue-double layer Amniotic membrane (AM) and evaluated the biomechanical properties and effects on corneal surface reconstruction in alkali-burned rabbit model. Biomechanic parameters were calculated by an electronic universal testing machine. Corneal alkali burning was done in the right eyes of thirty rabbits, which were randomized into three groups of ten animals each. The eyes in group 1 underwent fibrin glue-double layer AMT, the eyes in group 2 underwent ordinary single layer AMT, and the eyes in group 3 (control group) did not undergo any surgical procedure. Healing of corneal epithelial defect, extent of corneal vascularization and corneal clarity were assessed and compared at two time points. One month after surgery, animals were killed and the eyes were processed for histopathology. The fibrin glue-double layer AM composites had more ideal biomechanical properties. In fibrin glue-double layer AM group, the rate of epithelial healing, vascularization inhibition and corneal clarity was significantly better than the other two groups. Novel fibrin glue-double layer AMT with corneal alkali burns is more effective and useful for ocular surface reconstruction and has great potential applications.

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

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    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.

  18. Pulmonary phospholipids in amniotic fluid of pathologic pregnancies: relationship with clinical status of the newborn.

    Science.gov (United States)

    Zapata, A; Hernandez-Garcia, J M; Grande, C; Martinez, I; Perez, J; de la Fuente, P; Usandizaga, J A

    1989-06-01

    We evaluated phospholipids, C-peptide and cortisol levels in amniotic fluid of 203 pathologic pregnancies (63, class A, B and C diabetics; 11 class D, F and H diabetics; 44 preclampsia and 85 Rh-isoimmunization); the control group was 82 normal pregnant women. There was an acceleration of fetal pulmonary maturation in women with preclampsia and severe Rh-isoimmunization in class D, F and H diabetics (at 34 weeks gestation the incidence of mature surfactant (lecithin/sphingomyelin greater than or equal to 2.7 and presence of phosphatidyl-glycerol) in these groups was 30%, 50% and 100%, respectively, while it was zero in the control group). At 37 and 38 weeks only 44.4% of the class A, B and C diabetics had mature surfactant and there was a significant difference with respect to the control group (x2 = 4.9; p less than 0.05); C-peptide levels in these diabetics (class A, B and C) were higher than in controls (p less than 0.001); in pregnant women with accelerated fetal lung maturation they were lower. We demonstrated a close relationship between fetal pulmonary maturity and the type of surfactant in amniotic fluid, which was independent of gestational age.

  19. Reptile genomes open the frontier for comparative analysis of amniote development and regeneration.

    Science.gov (United States)

    Tollis, Marc; Hutchins, Elizabeth D; Kusumi, Kenro

    2014-01-01

    Developmental genetic studies of vertebrates have focused primarily on zebrafish, frog and mouse models, which have clear application to medicine and well-developed genomic resources. In contrast, reptiles represent the most diverse amniote group, but have only recently begun to gather the attention of genome sequencing efforts. Extant reptilian groups last shared a common ancestor ?280 million years ago and include lepidosaurs, turtles and crocodilians. This phylogenetic diversity is reflected in great morphological and behavioral diversity capturing the attention of biologists interested in mechanisms regulating developmental processes such as somitogenesis and spinal patterning, regeneration, the evolution of "snake-like" morphology, the formation of the unique turtle shell, and the convergent evolution of the four-chambered heart shared by mammals and archosaurs. The complete genome of the first non-avian reptile, the green anole lizard, was published in 2011 and has provided insights into the origin and evolution of amniotes. Since then, the genomes of multiple snakes, turtles, and crocodilians have also been completed. Here we will review the current diversity of available reptile genomes, with an emphasis on their evolutionary relationships, and will highlight how these genomes have and will continue to facilitate research in developmental and regenerative biology.

  20. Positive effects of bFGF modified rat amniotic epithelial cells transplantation on transected rat optic nerve.

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    Jia-Xin Xie

    Full Text Available Effective therapy for visual loss caused by optic nerve injury or diseases has not been achieved even though the optic nerve has the regeneration potential after injury. This study was designed to modify amniotic epithelial cells (AECs with basic fibroblast growth factor (bFGF gene, preliminarily investigating its effect on transected optic nerve.A human bFGF gene segment was delivered into rat AECs (AECs/hbFGF by lentiviral vector, and the gene expression was examined by RT-PCR and ELISA. The AECs/hbFGF and untransfected rat AECs were transplanted into the transected site of the rat optic nerve. At 28 days post transplantation, the survival and migration of the transplanted cells was observed by tracking labeled cells; meanwhile retinal ganglion cells (RGCs were observed and counted by employing biotin dextran amine (BDA and Nissl staining. Furthermore, the expression of growth associated protein 43 (GAP-43 within the injury site was examined with immunohistochemical staining.The AECs/hbFGF was proven to express bFGF gene and secrete bFGF peptide. Both AECs/hbFGF and AECs could survive and migrate after transplantation. RGCs counting implicated that RGCs numbers of the cell transplantation groups were significantly higher than that of the control group, and the AECs/hbFGF group was significantly higher than that of the AECs group. Moreover GAP-43 integral optical density value in the control group was significantly lower than that of the cell transplantation groups, and the value in the AECs/hbFGF group was significantly higher than that of the AECs group.AECs modified with bFGF could reduce RGCs loss and promote expression of GAP-43 in the rat optic nerve transected model, facilitating the process of neural restoration following injury.

  1. Amniotic Membrane Modifies the Genetic Program Induced by TGFß, Stimulating Keratinocyte Proliferation and Migration in Chronic Wounds.

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    Antonia Alcaraz

    Full Text Available Post-traumatic large-surface or deep wounds often cannot progress to reepithelialisation because they become irresponsive in the inflammatory stage, so intervention is necessary to provide the final sealing epidermis. Previously we have shown that Amniotic Membrane (AM induced a robust epithelialisation in deep traumatic wounds.To better understand this phenomenon, we used keratinocytes to investigate the effect of AM on chronic wounds. Using keratinocytes, we saw that AM treatment is able to exert an attenuating effect upon Smad2 and Smad3 TGFß-induced phosphorylation while triggering the activation of several MAPK signalling pathways, including ERK and JNK1, 2. This also has a consequence for TGFß-induced regulation on cell cycle control key players CDK1A (p21 and CDK2B (p15. The study of a wider set of TGFß regulated genes showed that the effect of AM was not wide but very concrete for some genes. TGFß exerted a powerful cell cycle arrest; the presence of AM however prevented TGFß-induced cell cycle arrest. Moreover, AM induced a powerful cell migration response that correlates well with the expression of c-Jun protein at the border of the healing assay. Consistently, the treatment with AM of human chronic wounds induced a robust expression of c-Jun at the wound border.The effect of AM on the modulation of TGFß responses in keratinocytes that favours proliferation together with AM-induced keratinocyte migration is the perfect match that allows chronic wounds to move on from their non-healing state and progress into epithelialization. Our results may explain why the application of AM on chronic wounds is able to promote epithelialisation.

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

    Science.gov (United States)

    Chang, Yu-Jen; Ho, Tsung-Yen; Wu, Mei-Ling; Hwang, Shiaw-Min; Chiou, Tzyy-Wen; Tsai, Ming-Song

    2013-01-01

    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.

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

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    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.

  4. 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.

  5. 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

  6. Evolution and Development of Ventricular Septation in the Amniote Heart

    Science.gov (United States)

    Poelmann, Robert E.; Groot, Adriana C. Gittenberger-de; Vicente-Steijn, Rebecca; Wisse, Lambertus J.; Bartelings, Margot M.; Everts, Sonja; Hoppenbrouwers, Tamara; Kruithof, Boudewijn P. T.; Jensen, Bjarke; de Bruin, Paul W.; Hirasawa, Tatsuya; Kuratani, Shigeru; Vonk, Freek; van de Put, Jeanne M. M. S.; de Bakker, Merijn A.; Richardson, Michael K.

    2014-01-01

    of this part of the vertebrate heart as well as understand the etiology of septal defects in human congenital heart malformations. PMID:25192012

  7. Evolution and development of ventricular septation in the amniote heart.

    Directory of Open Access Journals (Sweden)

    Robert E Poelmann

    evolution and development of this part of the vertebrate heart as well as understand the etiology of septal defects in human congenital heart malformations.

  8. 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.

  9. Effect of Orally Administered Morphine on the Development of Amniotic and Chorionic Cavities in Pregnant Wistar Rats

    Directory of Open Access Journals (Sweden)

    M Azarnia

    2010-12-01

    Full Text Available Introduction: Previous studies indicate that morphine consumption during pregnancy can inhibit embryonic development. The present study evaluated the effects of orally administered morphine on the development of embryonic cavities in nine day pregnant rats. Methods: Female Wistar rats weighing between 170-200 grams were used in the present study. Experimental group received morphine (0.05 mg/ml of tap water after one night coupling with male rats for mating. On the 9th day of pregnancy, the pregnant animals were anesthetized with chloroform and the embryos were removed surgically and fixed in 10% formalin for a week. The fixed embryos were processed and stained by H & E method and evaluated. The surface areas of amniotic and chorionic cavities and the whole body surface area of embryos were calculated using MOTIC software. Results: Findings indicated that both chorionic cavity and whole body surface areas of embryos were increased in the experimental group. In addition, surface area of amniotic cavity was reduced in experimental group. Conclusion: This study showed that oral morphine consumption caused a significant decrease in the embryonic amniotic cavity and furthermore, delay in natural embryonic amniotic cavity development can result in abnormal function and growth defect of the kidneys of infants of mothers addicted to morphine.

  10. Efficacy of moderate to severe ocular chemical injury and the acute phase treatment and amniotic membrane transplantation

    Directory of Open Access Journals (Sweden)

    Bing Bai

    2014-04-01

    Full Text Available AIM:To discuss effective ways of scientific treatment of ocular chemical injury by analyzing the effects of moderate to severe acute ocular chemical injury and amniotic membrane transplantation. METHODS:Totally 36 cases(43 eyesof Ⅱ degrees and above of ocular chemical injuries were collected, and clinical efficacy was observed by analyzing the causes of injury, the degree of injury, visual acuity and complications. RESULTS:The recovery of visual acuity was well in Ⅱ degrees of burns, and the acuity of 55.6% reached over 0.6. The recovery of visual acuity >0.3 in acid burn group was 80.0%, and the recovery of visual acuity >0.3 in alkali burn group was 56.0%. The difference between two groups was statistically significant(P0.3 in early amniotic membrane transplantation group was 69.7%, and in early no amniotic membrane transplantation group was 50.0%; the difference between two groups was statistically significant(PPPCONCLUSION: Early clinical treatment, and early amniotic membrane transplantation are very important for patients with ocular burn in acute phase to reduce complications and have a better recovery.

  11. 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.

  12. Prenatal diagnosis of megacystis-microcolon-intestinal hypoperistalsis syndrome: contribution of amniotic fluid digestive enzyme assay and fetal urinalysis.

    Science.gov (United States)

    Muller, Françoise; Dreux, Sophie; Vaast, Pascal; Dumez, Yves; Nisand, Israel; Ville, Yves; Boulot, Pierre; Guibourdenche, Jean; Althusser, Marc; Blin, Gérard; Gautier, Evelyne; Lespinard, Catherine; Perrotin, Franck; Poulain, Patrice; Sarramon, Marie-France

    2005-03-01

    Megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS) is a usually lethal disease during the first year of life. There is no specific ultrasound prenatal diagnosis and no identified genetic locus. The value of amniotic fluid digestive enzyme assay and fetal urine biochemistry in the prediction of MMIHS was analysed. Retrospective study of 14 MMIHS cases. Amniotic fluid digestive enzymes and fetal urine biochemistry were compared in MMIHS and megabladder (63 and 264 cases respectively). Abnormal amniotic fluid digestive enzyme profile (vomiting of bile or digestive secretion leakage) was observed in 8/10 MMIHS cases. These profiles were observed in 7/63 controls; 80% sensitivity (95%CI = 55%-100%); 89% specificity (95%CI = 81%-96%). Fetal urinalysis was normal in 12/12 MMIHS cases except high calcium (>0.6 mmol/l). This profile was observed in 33/264 megabladder control cases; 100% sensitivity; 98.7% specificity (95%CI = 83.5%-91.5%). For the first time, we propose a prenatal diagnosis of MMIHS based on amniotic fluid digestive enzyme assay and on fetal urinalysis. Copyright 2005 John Wiley & Sons, Ltd.

  13. 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

    Proteins found in amniotic fluid are mainly serum proteins, probably of maternal origin. About 5% of the total protein concentration has the potential of being fetal or decidual in origin. Only a few of these proteins have been isolated and characterised. In order to describe the foetal and decid...

  14. Are myo-inositol, glucose and zinc concentrations in amniotic fluid of fetuses with spina bifida different from controls?

    NARCIS (Netherlands)

    Groenen, P.; Wevers, R.A.; Janssen, F.; Tuerlings, J.H.A.M.; Merkus, J.M.W.M.; Steegers-Theunissen, R.P.M.

    2003-01-01

    OBJECTIVE: Associations are reported between myo-inositol, glucose, zinc and the occurrence of spina bifida. To gain more insight into the pathogenesis of spina bifida, the concentrations of myo-inositol, glucose and zinc were determined in amniotic fluids from pregnancies with a spina bifida or

  15. Intra amniotic administration and dietary inulin affect the iron status and intestinal functionality of iron deficient broiler chickens

    Science.gov (United States)

    Inulin, a linear beta-fructan, is present in a variety of plants, with relatively high levels of up to 20% in chicory root. It exhibits prebiotic properties and was shown to enhance mineral absorption. Our objectives were to assess the effect of intra-amniotic administration of inulin at 17d of incu...

  16. 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...

  17. The levels of the neutrophil elastase in the amniotic fluid of pregnant women whose infants develop bronchopulmonary dysplasia.

    Science.gov (United States)

    Ikeda, Sayako; Kihira, Kana; Yokoi, Akira; Tamakoshi, Koji; Miyazaki, Ken; Furuhashi, Madoka

    2015-03-01

    To clarify the association between amniotic neutrophil elastase levels and the development of bronchopulmonary dysplasia (BPD). The database between July 2001 and December 2012 was reviewed for women with amniocentesis on admission for amniotic fluid neutrophil elastase levels and with singleton deliveries between 22 + 0 and 31 + 6 weeks of gestation. Following deliveries, placentas were examined for histologic chorioamnionitis. The peripheral blood of the neonates was analyzed for acute phase reactants. Among 294 infants, no, mild, moderate or severe BPD was observed in 126, 89, 40 and 39 infants, respectively. The medians of gestational age on admission, at premature rupture of membranes and at delivery were significantly smaller in BPD (+) when compared with BPD (-) (p < 0.001). The median level of amniotic neutrophil elastase on admission was significantly greater in BPD (+) than that in BPD (-). Histologic chorioamnionitis and funisitis were both detected more frequently in BPD (+) patients than in BPD (-) patients. In a logistic regression model, the only variable that affected an increased chance of BPD was the gestational age at delivery (odds ratio, 0.58; 95% confidence interval, 0.36-0.92; p = 0.021). The level of amniotic neutrophil elastase cannot be a definitive risk factor for BPD.

  18. Prenatal sex hormones (maternal and amniotic fluid) and gender-related play behavior in 13-month-old Infants.

    NARCIS (Netherlands)

    Beek, C.; Goozen, S.H.M. van; Buitelaar, J.K.; Cohen-Kettenis, P.T.

    2009-01-01

    Testosterone, estradiol, and progesterone levels were measured in the second trimester of pregnancy in maternal serum and amniotic fluid, and related to direct observations of gender-related play behavior in 63 male and 63 female offspring at age 13 months. During a structured play session, sex

  19. 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

  20. Células-tronco do líquido amniótico Amniotic fluid stem cells

    Directory of Open Access Journals (Sweden)

    Sergio P. Bydlowski

    2009-05-01

    Full Text Available Desde o primeiro isolamento e cultivo de células-tronco embrionárias humanas, há mais de 10 anos, seu uso na pesquisa e terapia foi inibida por considerações éticas complexas e pelo risco de transformação maligna destas células indiferenciadas após transplante no paciente. As células-tronco adultas são eticamente aceitas e o risco de transformação maligna é muito baixo. Entretanto, seu potencial de diferenciação e sua capacidade proliferativa são limitados. Cerca de 6 anos atrás, a descoberta de célulastronco no líquido amniótico que expressavam Oct-4, um marcador específico de pluripotencialidade, com alta capacidade de proliferação e diferenciação, iniciou um novo campo promissor na área das células-tronco. Estas células têm potencial de se diferenciar em células dos três folhetos germinativos. Não formam tumores in vivo e não levantam os questionamentos éticos associados com as células-tronco embrionárias humanas. Futuras investigações revelarão se as células-tronco do líquido amniótico realmente irão representar um tipo intermediário com vantagens em relação tanto às células-tronco embrionárias quanto às adultas. Este artigo faz uma revisão acerca destes tópicos e das características biológicas das células-tronco do líquido amniótico.Since the first successful isolation and cultivation of human embryonic stem cells about 10 years ago, their use for research and therapy has been constrained by complex ethical considerations as well as by the risk of development of malignancies of undifferentiated embryonic stem cells after transplantation into the patient. Adult stem cells are ethically acceptable and the risk of tumor development is low. However, their differentiation potential and proliferative capacity are limited. About 6 years ago, the discovery of Oct-4 expressing amniotic fluid stem cells, a specific marker of pluripotency, with a high proliferative capacity, and multilineage

  1. miR-410 Inhibition Induces RPE Differentiation of Amniotic Epithelial Stem Cells via Overexpression of OTX2 and RPE65.

    Science.gov (United States)

    Choi, Soon Won; Kim, Jae-Jun; Seo, Min-Soo; Park, Sang-Bum; Kang, Tae-Wook; Lee, Jin Young; Lee, Byung-Chul; Kang, Insung; Shin, Tae-Hoon; Kim, Hyung-Sik; Yu, Kyung-Rok; Kang, Kyung-Sun

    2015-06-01

    The retinal pigment epithelium (RPE) is a highly specialized cell type located between the choroid and neural retina of the eye. RPE degeneration causes irreversible visual impairment, extending to blindness. Cell therapy has recently emerged as a potential therapeutic approach for retinal degeneration. MicroRNA-based differentiation of stem cells is a new strategy for producing tissue-specific cell types. In this study, we developed a novel microRNA-based strategy for RPE induction from human amniotic epithelial stem cells (AESCs). We identified microRNAs involved in RPE development in AESCs. Of 29 putative human RPE-relevant microRNAs, microRNA-410 (miR-410) was predicted to target multiple RPE development-relevant genes. Inhibition of miR-410 induces overexpression of immature and mature RPE-specific factors, including OTX2, RPE65, Bestrophin and EMMPRIN. These RPE-like cells were morphologically altered toward a cobblestone-like shape and were able to phagocytize microbeads. We showed that miR-410 directly regulates predicted target genes OTX2 and RPE65. Our microRNA-based strategy demonstrated RPE differentiation in AESCs by treatment of an antisense microRNA-410 (anti-miR-410), without the use of additional factors or exogenous transduction. These findings suggest that miR-410 inhibition can be a useful tool for directed cell differentiation and an attractive method for cell therapy in human retinal degenerative diseases.

  2. Corneal stromal acupuncture combined with amniotic membrane transplantation for treating bullous keratopathy

    Directory of Open Access Journals (Sweden)

    Chao-Qing Wang

    2014-06-01

    Full Text Available AIM: To investigate the clinical effect for treating bullous keratopathy(BKby anterior corneal stromal acupuncture combined with amniotic membrane transplantation.METHODS: Totally 35 patients(35 eyeswith bullous keratopathy were treated by corneal stromal acupuncture combined with amniotic membrane transplantation. All patients preoperative and postoperative underwent anterior segment OCT, corneal topography, corneal sensitivity and confocal microscopy. To observe postoperative ocular symptoms in patients with BK, recurrence of bulla, changes in corneal thickness, corneal sensitivity and changes in the organizational structure of the layers of the cornea.RESULTS: Thirty-five were followed up for 6-18mo. The symptom of pain disappeared in 32 cases(91%in the first day after operation and did not recur during follow-up. The symptom of pain relieved in 3 cases(9%in the first day after operation and disappeared in 3 days. Corneal epithelium of 9 cases(26%were all healed within 1wk, 21 cases(60%were all healed within 2wk, and 5 cases(14%were all healed within 3wk. Following up for 6-18mo, there was no recurrence of symptoms or bulla. A small amount of tiny bubbles in the surrounding area appeared in 2 cases after 3 and 4wk. All patients had no neovascularization, and had smooth corneal surface. The amnions of 30 cases(86%were thinning after 2mo, partially dissolved and absorped, not seen with the naked eye after 3mo. Thirty-four cases(97%had no changes in vision, one case(3%was from the light to front of the manual. After 2mo, corneal sensation decreased in 30 cases(86%, corneal thickness increased from preoperative 788±35μm to 940±43μm. After 12mo, corneal thickness increased to 1060±27μm. Results of confocal microscopy: after 3mo, the number density of the trigeminal nerve fibers under corneal basement membrane reduced, shallow stromal cells became into fibrotic stroma, deep stroma was more loose, and cells swelled significantly. The number of

  3. Physico-chemical properties of late-incubation egg amniotic fluid and a potential feed supplement

    Directory of Open Access Journals (Sweden)

    A. A. Omede

    2017-08-01

    Full Text Available Objective This study explored the physico-chemical properties of late-incubation egg amniotic fluid and a potential in ovo feed (IOF supplement. Methods Amniotic fluid was collected from broiler breeders (Ross 308, 51 weeks and Cobb 500, 35 weeks on day 17 after incubation. A mixture of high-quality soy protein supplement – Hamlet Protein AviStart (HPA was serially diluted in MilliQ water to obtain solutions ranging from 150 to 9.375 mg/mL. The mixtures were heat-treated (0, 30, 60 minutes in a waterbath (80°C and then centrifuged to obtain supernatants. The amniotic fluid and HPA supernatants were analysed for their physico-chemical properties. Results Only viscosity and K+ were significantly (p<0.05 different in both strains. Of all essential amino acids, leucine and lysine were in the highest concentration in both strains. The osmolality, viscosity and pCO2 of the supernatants decreased (p<0.05 with decreasing HPA concentration. Heat treatment significantly (p<0.05 affected osmolality, pH, and pCO2, of the supernatants. The interactions between HPA concentration and heat treatment were significant with regards to osmolality (p<0.01, pH (p<0.01, pCO2 (p<0.05, glucose (p<0.05, lactate (p<0.01 and acid-base status (p<0.01 of HPA solutions. The Ca2+, K+, glucose, and lactate increased with increasing concentration of HPA solution. The protein content of HPA solutions decreased (p<0.05 with reduced HPA solution concentrations. The supernatant from 150 mg/mL HPA solution was richest in glutamic acid, aspartic acid, arginine and lysine. Amino acids concentrations were reduced (p<0.05 with each serial dilution but increased with longer heating. Conclusion The values obtained in the primary solution (highest concentration are close to the profiles of high-protein ingredients. This supplement, as a solution, hence, may be suitable for use as an IOF supplement and should be tested for this potential.

  4. Reprogramming human amniotic fluid stem cells to functional pluripotency by manipulation of culture conditions.

    OpenAIRE

    sprotocols

    2015-01-01

    Authors: Dafni Moschidou & Pascale V Guillot ### Abstract Pluripotent stem cells have potential applications in regenerative medicine, disease modelling and drug screening. Induced pluripotent stem (iPS) cells have first been generated from fibroblasts using retroviral insertion of OCT4A, SOX2, c-MYC and KLF4. Since then, a number of methods have been developed to avoid the random integration of ectopic factors in the genome and the low efficiency of the process. Those include alt...

  5. Evaluation of repair in duodenal perforation with human amniotic membrane: An animal model (dog

    Directory of Open Access Journals (Sweden)

    Leila Ghahramani

    2014-01-01

    Conclusion: It seems that repairing duodenal wall defect with HAM would result in better histological outcomes compared with what is seen in simple duodenoraphy in animal models. However, there is no significant difference regarding surgical findings.

  6. Immune activation in amniotic fluid from Zika virus-associated microcephaly.

    Science.gov (United States)

    Ornelas, Alice M M; Pezzuto, Paula; Silveira, Paola P; Melo, Fabiana O; Ferreira, Thales A; Oliveira-Szejnfeld, Patricia S; Leal, Jeime I; Amorim, Melania M R; Hamilton, Stuart; Rawlinson, William D; Cardoso, Cynthia C; Nixon, Douglas F; Tanuri, Amilcar; Melo, Adriana S; Aguiar, Renato S

    2017-01-01

    Recent advances in the understanding of neuropathogenesis associated with Zika virus (ZIKV) infection has led to descriptions of neonatal microcephaly cases. However, none of these reports have evaluated the humoral response during ZIKV infection. We report here polyfunctional immune activation associated with increased interferon-gamma-inducible protein 10, interleukin (IL)-6, IL-8, vascular endothelial growth factor (VEGF), monocyte chemoattractive protein 1 (MCP-1), and granulocyte colony-stimulating factor (G-CSF) levels in the amniotic fluid of ZIKV-positive pregnant women with neonatal microcephaly. These cytokines have been associated not only with neuronal damage, but also with differentiation and proliferation of neural progenitor cells. Our results suggested that the immune activation caused by ZIKV infection in the uterine environment could also interfere with fetal development. ANN NEUROL 2017;81:152-156. © 2016 American Neurological Association.

  7. '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

  8. Trace element determination in amniotic fluid by total reflection X-ray fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Greaves, E.D.; Sajo-Bohus, L.; Castelli, C.; Borgerg, C. [Universidad Simon Bolivar, Caracas (Venezuela); Meitin, J.; Liendo, J.

    1995-03-01

    A new method is reported for the determination of Fe, Cu, Zn, and Br in amniotic fluid (AF) by Total Reflection X-Ray Fluorescence. The irradiation of AF samples with monochromatic X-Rays reduces the scattering background from the organic matrix and avoids the need for sample digestion. Sample manipulation is reduced to centrifuging and adding cobalt as internal standard. Lower detection limits obtained are 109, 53, 44 and 37 ppb for Fe, Cu, Zn and Br respectively. Measurement precision depends on element concentrations and can be as low as 1.5% SD. Results of the analysis of 34 AF samples from Venezuelan pregnant patients agree with previously reported ranges of Fe, Cu and Zn. Other elements observed but not quantified are Cl, K, Ca in all spectra and Pb and Sr in some of them. (author).

  9. Rh alloimmunization: Doppler or amniotic fluid analysis in the prediction of fetal anemia?

    Science.gov (United States)

    Nardozza, Luciano Marcondes Machado; Moron, Antonio Fernandes; Araujo Júnior, Edward; Camano, Luiz; Chinen, Paulo Alexandre; Torloni, Maria Regina

    2007-02-01

    Compare the results of amniocentesis and those of middle cerebral artery peak systolic velocity to detect anemic fetuses in Rh alloimmunized pregnancies. Descriptive observational study involving 99 consecutive Rh-negative pregnancies: 74 alloimmunized patients submitted to amniotic fluid spectrophotometry (group 1) and 25 alloimmunized cases managed with Doppler ultrasonography (group 2). Variables analyzed were the following: need for neonatal transfusion and neonatal hematocrit. Neonatal hematocrit was significantly lower and the need for neonatal transfusion was significantly higher in cases managed with spectrophotometry compared to those managed with Doppler. Infants managed with Doppler velocimetry had higher hematocrit and less need for neonatal transfusion, suggesting that this non-invasive method of monitoring fetal anemia may best preserve the neonate.

  10. Combined Conjunctival Autograft and Overlay Amniotic Membrane Transplantation; a Novel Surgical Treatment for Pterygium

    Directory of Open Access Journals (Sweden)

    Siamak Zarei Ghanavati

    2014-01-01

    Full Text Available The authors report the long-term results of combined conjunctival autograft and overlay amniotic membrane transplantation (AMT for treatment of pterygium as a new surgical technique. Nineteen patients including 12 male and 7 female subjects with pterygium (primary, 14 cases; recurrent, 5 cases underwent combined conjunctival autograft and overlay AMT and were followed from 10 to 26 months. Mean age was 44.21±12.49 (range, 29.0-73.0 years. In one patient with grade T3 primary pterygium, the lesion recurred (5.2%, recurrence rate. No intra-and postoperative complication developed. This procedure seems a safe and effective surgical technique for pterygium treatment. Protection of the ocular surface during the early postoperative period reduces the friction-induced inflammation and might be helpful to prevent the recurrence.

  11. Combined conjunctival autograft and overlay amniotic membrane transplantation; a novel surgical treatment for pterygium.

    Science.gov (United States)

    Ghanavati, Siamak Zarei; Shousha, Mohamed Abou; Betancurt, Carolina; Perez, Victor L

    2014-01-01

    The authors report the long-term results of combined conjunctival autograft and overlay amniotic membrane transplantation (AMT) for treatment of pterygium as a new surgical technique. Nineteen patients including 12 male and 7 female subjects with pterygium (primary, 14 cases; recurrent, 5 cases) underwent combined conjunctival autograft and overlay AMT and were followed from 10 to 26 months. Mean age was 44.21±12.49 (range, 29.0-73.0) years. In one patient with grade T3 primary pterygium, the lesion recurred (5.2%, recurrence rate). No intra-and postoperative complication developed. This procedure seems a safe and effective surgical technique for pterygium treatment. Protection of the ocular surface during the early postoperative period reduces the friction-induced inflammation and might be helpful to prevent the recurrence.

  12. 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.

  13. Amniotic Fluid Chemokines and Autism Spectrum Disorders: An Exploratory Study Utilizing a Danish Historic Birth Cohort

    DEFF Research Database (Denmark)

    Abdallah, Morsi; Larsen, Nanna; Grove, Jakob

    2012-01-01

    Introduction: 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. Material...... controls was sufficient for Luminex analysis. Including all individuals in the cohort yielded no significant differences in chemokine levels in cases versus controls. Logistic regression analyses, performed on individuals diagnosed using ICD-10 only, showed increased risk for ASD with elevated MCP-1....... Case-control differences were assessed as dichotomized at below the 10th percentile or above the 90th percentile cut-off points derived from the control biomarker distributions (logistic regression) or continuous measures (tobit regression). Results and Conclusion AF volume for 331 cases and 698...

  14. Application of the amniotic membrane extract (AMX) for the persistent epithelial defect (PED) of the cornea.

    Science.gov (United States)

    Kordić, Rajko; Suić, Smiljka Popović; Jandroković, Sonja; Kalauz, Miro; Kuzman, Tomislav; Skegro, Ivan; Jukić, Tomislav

    2013-04-01

    A lot of pathological conditions could provoke damaging of the innervations of the cornea and lead to persistent epithelial defect (PED). AMX is lyophilized preparation of amniotic membrane (AM), which contains biological components and efficacy of AM for treatment of the corneal surface defects. In processing all the substances essential for biological effects of AM are preserved (growth factors, neutrophins, interleukins, receptors, fibronectins and different types of collagen). A patient can apply AMX as eye drops by himself in his home, thus avoiding surgical procedure. We presented two patients with PED; we treated them with eye drops of AMX, 2 drops every hour during day time. There was observed healing effect by reducing epithelial defect. Almost after a second day of application, and after 1-2 week period PED that persisted for weeks healed completely.

  15. [Karyotype analysis of amniotic fluid cells and comparison of chromosomal abnormality rate during second trimester].

    Science.gov (United States)

    Zhang, Yue-ping; Wu, Jun-ping; Li, Xiao-tian; Lei, Cai-xia; Xu, Jian-Zhong; Yin, Min

    2011-09-01

    To investigate the karyotypes of amniotic fluid cells and compare the incidence of chromosomal abnormality as well as to evaluate the clinical significance of abnormal karyotypes. A total of 13 648 pregnant women came to Shanghai Jiai Genetics and IVF Institute, Obstetrics and Gynecology Hospital, Fudan University to do amniocentesis from September 1998 to November 2010, and 13 795 amniotic fluid specimens were successfully extracted and cultured, thus 13 795 fetuses received karyotype diagnosis. These fetuses were grouped according to different indications. If maternal age was ≥ 35, the fetuses were grouped into the advanced maternal age group (4065); and if maternal serum screening test revealed high-risk of trisomy 18 or trisomy 21, the fetuses were grouped into the high-risk serum screening group (6462); and those with abnormal signs of ultrasound screening were grouped into the abnormal ultrasound signs group (1539); and if either of the parents was with chromosome abnormalities, the fetus was grouped into the paternal/maternal abnormality group (108); whereas the remainder were grouped in other factors group (1621). The amniotic fluid cells were in-situ cultured on coverslips, harvested by conventional G-banded methods, and then analyzed by two doctors. In order to get rapid diagnosis, some pregnant women whose gestational age ≥ 26 weeks accepted fluorescense in situ hybridization (FISH). FISH was done on 78 uncultured amniotic fluid specimens using probes located at chromosome 13, 18, 21, X, Y. Some parents were required to analyze lymphocyte karyotype to help judging the origin of abnormal karyotype. (1) Classification and composition of abnormal karyotypes in each group: a total of 388 abnormal karyotypes were found among 13 795 fetuses, and the abnormal rate was 2.813% (388/13 795). Of the 388 fetuses, aneuploidy was the most common pattern which was up to 59.8% (232/388); autosomal structural abnormality rate was 24.7% (96/388); mosaicism was 12

  16. Prenatal detection of trisomy 21 and 18 from amniotic fluid by quantitative fluorescent polymerase chain reaction.

    Science.gov (United States)

    Tóth, T; Findlay, I; Papp, C; Tóth-Pál, E; Marton, T; Nagy, B; Quirke, P; Papp, Z

    1998-01-01

    Prenatal diagnosis of fetal trisomies is usually performed by cytogenetic analysis on amniotic fluid. This requires lengthy laboratory procedures and high costs, and is unsuitable for large scale screening of pregnant women. An alternative method, which is both rapid and inexpensive and suitable for diagnosing trisomies even from single fetal cells, is the fluorescent polymerase chain reaction using polymorphic small tandem repeats (STRs). In this paper we present the preliminary results of a larger study comparing parallel prenatal diagnoses of trisomies 21 and 18 using cytogenetics with quantitative fluorescent polymerase chain reaction using STR markers. The results obtained by the two techniques were concordant in all cases. This is the first study reporting significant numbers of prenatal diagnoses using the quantitative fluorescent polymerase chain reaction. We believe that further studies on greater numbers of samples will determine the absolute reliability of this technique. These results also provide a model for diagnosis of trisomy from single fetal cells isolated from maternal blood. PMID:9507392

  17. Minimally Invasive Pterygium Surgery: Sutureless Excision with Amniotic Membrane and Hydrogel Sealant

    Directory of Open Access Journals (Sweden)

    Sailaja Bondalapati

    2016-02-01

    Full Text Available Purpose: To describe a novel technique for sutureless pterygium surgery using ReSure® tissue sealant. Methods: In this retrospective observational case series, we describe a modified procedure for pterygium excision followed by amniotic membrane transplant (AMT adhered to the corneal and conjunctival defects using ReSure tissue sealant. Results: Nine eyes of seven patients (age range: 28-80 years, 4 females and 3 males underwent pterygium removal with AMT followed by adherence of tissue to the conjunctival edges with ReSure. No issues with transplant dislocation or failure and no intra- or postoperative complications were noted. No recurrences were noted during the follow-up period. Conclusion: ReSure may be considered as a potential sealant to adhere AMT to defective corneal and conjunctival tissues in sutureless pterygium surgery.

  18. Morphological changes in the placenta and decidua after induction of abortion by extra-amniotic prostaglandin.

    Science.gov (United States)

    Fox, H; Herd, M E; Harilal, K R

    1978-03-01

    Seventeen placentae from second trimester abortions induced by the injection of extra-amniotic prostaglandin have been examined. The extraplacental membranes, the chorionic plate of the placenta, the placental intervillous space and the placental villi were normal in all cases. In all but one specimen lesions were, however, noted in the decidua, these including focal decidual necrosis, intradecidual haemorrhage, thrombosis of maternal vessels and dilatation and congestion of the maternal vasculature. Similar decidual lesions were also seen in placentae from spontaneous and surgical second trimester abortions and it is concluded that prostaglandins do not produce any specific lesions in the placenta or decidua. It is suggested that the negative findings of this study indicate that prostaglandins do not induce abortion by either direct injury to the placenta or by provoking utero-placental ischaemia.

  19. Phacoemulsification after penetrating keratoplasty with autologous limbal transplant and amniotic membrane transplant in chemical burns

    Directory of Open Access Journals (Sweden)

    Arora Ritu

    2005-01-01

    Full Text Available We report a patient who had earlier penetrating keratoplasty with amniotic membrane transplant and autologous limbal cell transplant for chemical injury who underwent cataract surgery by phacoaspiration. A posterior limbal incision with corneal valve was made superotemporally with extreme caution to avoid damage to the limbal graft. Aspiration flow rates and vacuum were kept low to avoid any turbulence during surgery. A 6.0 mm optic diameter acrylic foldable intraocular lens was inserted in the bag. The patient achieved a best-corrected visual acuity of 6/12 at 10 months′ follow-up with a clear corneal graft. We conclude that caution during wound construction and phacoaspiration can help preserve corneal and limbal graft integrity in patients undergoing cataract surgery after corneal graft and limbal transplantation.

  20. The use of Amniotic membrane in the treatment of Burns in Children a clinical trial at the university Teaching Hospital, Lusaka

    CERN Document Server

    Katebe, K R

    1995-01-01

    This is a clinical trial which was carried out at the University Teaching Hospital, Lusaka from the 1st of July to the 31st December, 1994. It involved treatment of burns in forty children using gamma irradiated amniotic membrane produced at the hospital. The results showed that it is feasible to produce Gamma irradiated biological dressings from amniotic membrane at this hospital. The amniotic membrane was easy to apply on burns and the treatment was acceptable to the majority of parents with burnt children. The use of amniotic membrane was non inflammatory to the wounds in all forty patients (100%), reduced wound infection in thirty three patients (82.5%), increased the rate of wound healing in thirty nine patients (97.5%), and resulted in good quality wound healing in thirty one patients (77.5%). Therefore, the treatment offers a good alternative in the treatment of burns in children at the hospital

  1. [Alpha-foetoprotein assessment in amniotic fluid for the detection of neural tube defects: limited added value beyond week 20 ultrasound; retrospective study

    NARCIS (Netherlands)

    Kooper, A.J.A.; Smits, A.P.T.; Feuth, A.B.; Burgt, I. van der; Zondervan, H.A.; Quartero, R.W.P.; Boekkooi, P.F.; Rijnders, R.J.P.; Creemers, J.W.T.; Thomas, C.M.G.

    2008-01-01

    OBJECTIVE: To evaluate the diagnostic additional value of routine alpha-foetoprotein (AFP) assessment in amniotic fluid for the detection of neural tube defects (NTDs), compared with week 20 ultrasonographic examination. DESIGN: Retrospective. METHOD: We retrospectively determined AFP concentrations

  2. AMNIOTIC FLUID LAMELLAR BODY COUNT FOR PREDICTION OF FOETAL LUNG MATURITY

    Directory of Open Access Journals (Sweden)

    Aruna Ratikrinda

    2016-06-01

    Full Text Available INTRODUCTION Predicting maturity of the foetal lung is extremely important in many obstetric situations as respiratory distress syndrome caused by surfactant deficiency remains one of the leading causes of neonatal morbidity and mortality. The need for predicting the foetal lung maturity by means of an accurate test which is done rapidly and available in majority of centres was the objective for this study. AIM To find out the optimum lamellar body count which correlates with foetal lung maturity and to compare the lamellar body count with shake bubble test. MATERIAL AND METHODS This prospective study was designed to evaluate the lamellar body count by standard haematology cell counter that is coulter counter and compare it with shake bubble test. Both the tests were done on 100 amniotic fluid samples in women with gestational age more than 28 weeks. Patients recruited were those who were sure of their last menstrual period (LMP, who were in active labour, and likely to deliver within 72 hours of collection of sample. Amniotic fluid samples obtained by transabdominal amniocentesis or by aspirating the forewaters per vaginum. Both shake bubble test and lamellar body count were done on the sample. RESULTS Lamellar body counts ranged from 10,000-2,43,000/microlitre and had a linear relationship with gestational age. The cut-off level of 30,000/microlitre was considered to be optimum to predict foetal lung maturity. Sensitivity and negative predictive value of lamellar body count at 30,000/microlitre and shake bubble test were comparable at 91.7%/91.7% and 97.3/96.6% respectively whereas with respect to specificity and positive predictive value, lamellar body count was superior to shake bubble test at 93.4%/73.7% and 81.5%/50%. CONCLUSION Lamellar body count is a rapid, inexpensive, simple and more reliable test to assess foetal lung maturity

  3. A comparative examination of odontogenic gene expression in both toothed and toothless amniotes.

    Science.gov (United States)

    Lainoff, Alexis J; Moustakas-Verho, Jacqueline E; Hu, Diane; Kallonen, Aki; Marcucio, Ralph S; Hlusko, Leslea J

    2015-05-01

    A well-known tenet of murine tooth development is that BMP4 and FGF8 antagonistically initiate odontogenesis, but whether this tenet is conserved across amniotes is largely unexplored. Moreover, changes in BMP4-signaling have previously been implicated in evolutionary tooth loss in Aves. Here we demonstrate that Bmp4, Msx1, and Msx2 expression is limited proximally in the red-eared slider turtle (Trachemys scripta) mandible at stages equivalent to those at which odontogenesis is initiated in mice, a similar finding to previously reported results in chicks. To address whether the limited domains in the turtle and the chicken indicate an evolutionary molecular parallelism, or whether the domains simply constitute an ancestral phenotype, we assessed gene expression in a toothed reptile (the American alligator, Alligator mississippiensis) and a toothed non-placental mammal (the gray short-tailed opossum, Monodelphis domestica). We demonstrate that the Bmp4 domain is limited proximally in M. domestica and that the Fgf8 domain is limited distally in A. mississippiensis just preceding odontogenesis. Additionally, we show that Msx1 and Msx2 expression patterns in these species differ from those found in mice. Our data suggest that a limited Bmp4 domain does not necessarily correlate with edentulism, and reveal that the initiation of odontogenesis in non-murine amniotes is more complex than previously imagined. Our data also suggest a partially conserved odontogenic program in T. scripta, as indicated by conserved Pitx2, Pax9, and Barx1 expression patterns and by the presence of a Shh-expressing palatal epithelium, which we hypothesize may represent potential dental rudiments based on the Testudinata fossil record. © 2015 Wiley Periodicals, Inc.

  4. 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.

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

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    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

  6. Phosphatidylglycerol in amniotic fluid. Comparison of an "ultrasensitive" immunologic assay with TLC and enzymatic assay.

    Science.gov (United States)

    Eisenbrey, A B; Epstein, E; Zak, B; McEnroe, R J; Artiss, J D; Kiechle, F L

    1989-03-01

    Phosphatidylglycerol (PG) in amniotic fluid is recognized as a good indicator of fetal lung maturity and is unaffected by moderate amounts of blood or meconium contamination. A rapid immunologic agglutination assay, Ultrasensitive AmnioStat-FLM (FLM), was compared with two-dimensional thin-layer chromatography (TLC) and an enzymic, colorimetric procedure (E-PG). Eighty amniotic fluid specimens were analyzed. FLM results were reported as high (H), intermediate (I), or low positive (L). TLC was compared with FLM:H (n = 27), mean 0.14 (fraction of total phospholipids); I (n = 7), mean 0.11; L (n = 9), mean 0.03; negative results had no detectable PG by TLC. In 33 cases E-PG was compared with FLM:H (n = 9), mean 7.0 mumol/L; I (n = 5), mean 8.1 mumol/L; L (n = 3), mean 3.0 mumol/L; negative (n = 16), mean 3.2 mumol/L. Records were reviewed in 70 cases. Thirty cases were excluded: sample to delivery time was greater than 72 hours; steroids were given or sepsis was documented. Fetal lung immaturity was clinically present in six cases: respiratory distress syndrome in three cases and transient tachypnea of the newborn (TTN) in three cases. One false positive result was identified (TTN, FLM:H). FLM sensitivity for fetal lung maturity was 85.3%, specificity was 83.3%, and the positive predictive value for fetal lung maturity was 96.7%. FLM is a fast, reliable indicator of fetal lung maturity.

  7. Lactate in Amniotic Fluid: Predictor of Labor Outcome in Oxytocin-Augmented Primiparas' Deliveries.

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    Eva Wiberg-Itzel

    Full Text Available One of the major complications related to delivery is labor dystocia, or an arrested labor progress. Many dystocic deliveries end vaginally after administration of oxytocin, but a large numbers of women with labor dystocia will undergo a long and unsafe parturition. As a result of the exertion required in labor, the uterus produces lactate. The uterine production of lactate is mirrored by the level of lactate in amniotic fluid (AFL.To evaluate whether the level of AFL, analysed in a sample of amniotic fluid collected vaginally at arrested labor when oxytocin was needed, could predict labor outcome in nulliparous deliveries.A prospective multicentre study including 3000 healthy primiparous women all with a singleton pregnancy, gestational age 37 to 42 weeks and no maternal /fetal chronic and/or pregnancy-related conditions. A spontaneous onset of labor, regular contractions and cervical dilation ≥ 3 cm were required before the women were invited to take part in the study.AFL, analysed within 30 minutes before augmentation, provides information about delivery outcome. Sensitivity for an acute cesarean section according to high (≥10.1mmol/l or low (12h (p = 0.04, post-partum fever (>38°C, p = 0.01 and post-partum haemorrhage >1.5L (p = 0.04.The AFL is a good predictor of delivery outcome in arrested nulliparous deliveries. Low levels of AFL may support the decision to continue a prolonged vaginal labor by augmentation with oxytocin. A high level of AFL correlates with operative interventions and post-partum complications.

  8. 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

    2016-01-01

    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) fetal inflammatory response syndrome (FIRS). 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. 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 identification of FIRS in a population with a prevalence of 20%. Acute histologic chorioamnionitis and funisitis are associated with intra-amniotic infection and the presence of FIRS. However, current pathologic methods have limitations in the identification of the fetus exposed to microorganisms present in the amniotic cavity. Further studies are thus required to determine whether molecular markers can enhance the performance

  9. Bupropion therapy during pregnancy: the drug and its major metabolites in umbilical cord plasma and amniotic fluid.

    Science.gov (United States)

    Fokina, Valentina M; West, Holly; Oncken, Cheryl; Clark, Shannon M; Ahmed, Mahmoud S; Hankins, Gary D V; Nanovskaya, Tatiana N

    2016-10-01

    Bupropion is used for treatment of depression during pregnancy. However, its use as a smoking cessation aid for pregnant women is currently under evaluation. The aim of this opportunistic study was to investigate the transfer of bupropion and its major pharmacologically active metabolites, hydroxybupropion and threohydrobupropion, across the placenta in vivo. In addition, the concentrations of the drug and its metabolites were determined in the amniotic fluid. The following samples were collected at deliveries from 22 women taking bupropion: maternal blood (n = 22), umbilical cord venous blood (n = 22), and amniotic fluid (n = 9). The concentrations of the drug and its metabolites in blood plasma and amniotic fluid were determined by means of liquid chromatography-mass spectrometry. Placental passage was calculated as a ratio of umbilical cord venous plasma to maternal plasma concentrations. The levels of hydroxybupropion and threohydrobupropion in umbilical cord venous plasma were invariably lower than their corresponding concentrations in maternal plasma. The concentrations of bupropion in umbilical cord plasma were lower than in maternal plasma in the majority of the maternal-cord blood pairs. The median values of the umbilical cord venous plasma to maternal plasma ratios were: bupropion, 0.53 (interquartile range 0.35, n = 18), hydroxybupropion, 0.21 (interquartile range 0.12, n = 18), and threohydrobupropion, 0.61 (interquartile range 0.11, n = 21). In umbilical cord venous plasma, the median concentration of bupropion was 5.3 ng/mL; hydroxybupropion, 103.6 ng/mL; and threohydrobupropion, 59.6 ng/mL. Bupropion and its metabolites were detectable in the amniotic fluid but the concentrations of threohydrobupropion were higher than those in the corresponding umbilical cord venous plasma. Bupropion and its active metabolites cross the placenta to the fetal circulation. The concentrations of hydroxybupropion and threohydrobupropion in umbilical cord

  10. The role of the multiple banded antigen of Ureaplasma parvum in intra-amniotic infection: major virulence factor or decoy?

    Directory of Open Access Journals (Sweden)

    Samantha J Dando

    Full Text Available The multiple banded antigen (MBA is a predicted virulence factor of Ureaplasma species. Antigenic variation of the MBA is a potential mechanism by which ureaplasmas avoid immune recognition and cause chronic infections of the upper genital tract of pregnant women. We tested whether the MBA is involved in the pathogenesis of intra-amniotic infection and chorioamnionitis by injecting virulent or avirulent-derived ureaplasma clones (expressing single MBA variants into the amniotic fluid of pregnant sheep. At 55 days of gestation pregnant ewes (n = 20 received intra-amniotic injections of virulent-derived or avirulent-derived U. parvum serovar 6 strains (2×10⁴ CFU, or 10B medium (n = 5. Amniotic fluid was collected every two weeks post-infection and fetal tissues were collected at the time of surgical delivery of the fetus (140 days of gestation. Whilst chronic colonisation was established in the amniotic fluid of animals infected with avirulent-derived and virulent-derived ureaplasmas, the severity of chorioamnionitis and fetal inflammation was not different between these groups (p>0.05. MBA size variants (32-170 kDa were generated in vivo in amniotic fluid samples from both the avirulent and virulent groups, whereas in vitro antibody selection experiments led to the emergence of MBA-negative escape variants in both strains. Anti-ureaplasma IgG antibodies were detected in the maternal serum of animals from the avirulent (40% and virulent (55% groups, and these antibodies correlated with increased IL-1β, IL-6 and IL-8 expression in chorioamnion tissue (p<0.05. We demonstrate that ureaplasmas are capable of MBA phase variation in vitro; however, ureaplasmas undergo MBA size variation in vivo, to potentially prevent eradication by the immune response. Size variation of the MBA did not correlate with the severity of chorioamnionitis. Nonetheless, the correlation between a maternal humoral response and the expression of chorioamnion

  11. Exposure to intrauterine inflammation alters metabolomic profiles in the amniotic fluid, fetal and neonatal brain in the mouse.

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    Amy G Brown

    Full Text Available Exposure to prenatal inflammation is associated with diverse adverse neurobehavioral outcomes in exposed offspring. The mechanism by which inflammation negatively impacts the developing brain is poorly understood. Metabolomic profiling provides an opportunity to identify specific metabolites, and novel pathways, which may reveal mechanisms by which exposure to intrauterine inflammation promotes fetal and neonatal brain injury. Therefore, we investigated whether exposure to intrauterine inflammation altered the metabolome of the amniotic fluid, fetal and neonatal brain. Additionally, we explored whether changes in the metabolomic profile from exposure to prenatal inflammation occurs in a sex-specific manner in the neonatal brain.CD-1, timed pregnant mice received an intrauterine injection of lipopolysaccharide (50 μg/dam or saline on embryonic day 15. Six and 48 hours later mice were sacrificed and amniotic fluid, and fetal brains were collected (n = 8/group. Postnatal brains were collected on day of life 1 (n = 6/group/sex. Global biochemical profiles were determined using ultra performance liquid chromatography/tandem mass spectrometry (Metabolon Inc.. Statistical analyses were performed by comparing samples from lipopolysaccharide and saline treated animals at each time point. For the P1 brains, analyses were stratified by sex.Exposure to intrauterine inflammation induced unique, temporally regulated changes in the metabolic profiles of amniotic fluid, fetal brain and postnatal brain. Six hours after exposure to intrauterine inflammation, the amniotic fluid and the fetal brain metabolomes were dramatically altered with significant enhancements of amino acid and purine metabolites. The amniotic fluid had enhanced levels of several members of the (hypo xanthine pathway and this compound was validated as a potential biomarker. By 48 hours, the number of altered biochemicals in both the fetal brain and the amniotic fluid had declined, yet unique

  12. The Role of the Multiple Banded Antigen of Ureaplasma parvum in Intra-Amniotic Infection: Major Virulence Factor or Decoy?

    Science.gov (United States)

    Dando, Samantha J.; Nitsos, Ilias; Kallapur, Suhas G.; Newnham, John P.; Polglase, Graeme R.; Pillow, J. Jane; Jobe, Alan H.; Timms, Peter; Knox, Christine L.

    2012-01-01

    The multiple banded antigen (MBA) is a predicted virulence factor of Ureaplasma species. Antigenic variation of the MBA is a potential mechanism by which ureaplasmas avoid immune recognition and cause chronic infections of the upper genital tract of pregnant women. We tested whether the MBA is involved in the pathogenesis of intra-amniotic infection and chorioamnionitis by injecting virulent or avirulent-derived ureaplasma clones (expressing single MBA variants) into the amniotic fluid of pregnant sheep. At 55 days of gestation pregnant ewes (n = 20) received intra-amniotic injections of virulent-derived or avirulent-derived U. parvum serovar 6 strains (2×104 CFU), or 10B medium (n = 5). Amniotic fluid was collected every two weeks post-infection and fetal tissues were collected at the time of surgical delivery of the fetus (140 days of gestation). Whilst chronic colonisation was established in the amniotic fluid of animals infected with avirulent-derived and virulent-derived ureaplasmas, the severity of chorioamnionitis and fetal inflammation was not different between these groups (p>0.05). MBA size variants (32–170 kDa) were generated in vivo in amniotic fluid samples from both the avirulent and virulent groups, whereas in vitro antibody selection experiments led to the emergence of MBA-negative escape variants in both strains. Anti-ureaplasma IgG antibodies were detected in the maternal serum of animals from the avirulent (40%) and virulent (55%) groups, and these antibodies correlated with increased IL-1β, IL-6 and IL-8 expression in chorioamnion tissue (p<0.05). We demonstrate that ureaplasmas are capable of MBA phase variation in vitro; however, ureaplasmas undergo MBA size variation in vivo, to potentially prevent eradication by the immune response. Size variation of the MBA did not correlate with the severity of chorioamnionitis. Nonetheless, the correlation between a maternal humoral response and the expression of chorioamnion cytokines is a

  13. The role of the multiple banded antigen of Ureaplasma parvum in intra-amniotic infection: major virulence factor or decoy?

    Science.gov (United States)

    Dando, Samantha J; Nitsos, Ilias; Kallapur, Suhas G; Newnham, John P; Polglase, Graeme R; Pillow, J Jane; Jobe, Alan H; Timms, Peter; Knox, Christine L

    2012-01-01

    The multiple banded antigen (MBA) is a predicted virulence factor of Ureaplasma species. Antigenic variation of the MBA is a potential mechanism by which ureaplasmas avoid immune recognition and cause chronic infections of the upper genital tract of pregnant women. We tested whether the MBA is involved in the pathogenesis of intra-amniotic infection and chorioamnionitis by injecting virulent or avirulent-derived ureaplasma clones (expressing single MBA variants) into the amniotic fluid of pregnant sheep. At 55 days of gestation pregnant ewes (n = 20) received intra-amniotic injections of virulent-derived or avirulent-derived U. parvum serovar 6 strains (2×10⁴ CFU), or 10B medium (n = 5). Amniotic fluid was collected every two weeks post-infection and fetal tissues were collected at the time of surgical delivery of the fetus (140 days of gestation). Whilst chronic colonisation was established in the amniotic fluid of animals infected with avirulent-derived and virulent-derived ureaplasmas, the severity of chorioamnionitis and fetal inflammation was not different between these groups (p>0.05). MBA size variants (32-170 kDa) were generated in vivo in amniotic fluid samples from both the avirulent and virulent groups, whereas in vitro antibody selection experiments led to the emergence of MBA-negative escape variants in both strains. Anti-ureaplasma IgG antibodies were detected in the maternal serum of animals from the avirulent (40%) and virulent (55%) groups, and these antibodies correlated with increased IL-1β, IL-6 and IL-8 expression in chorioamnion tissue (p<0.05). We demonstrate that ureaplasmas are capable of MBA phase variation in vitro; however, ureaplasmas undergo MBA size variation in vivo, to potentially prevent eradication by the immune response. Size variation of the MBA did not correlate with the severity of chorioamnionitis. Nonetheless, the correlation between a maternal humoral response and the expression of chorioamnion cytokines is a

  14. 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.

  15. Moderate postnatal hyperoxia accelerates lung growth and attenuates pulmonary hypertension in infant rats after exposure to intra-amniotic endotoxin.

    Science.gov (United States)

    Tang, Jen-Ruey; Seedorf, Gregory J; Muehlethaler, Vincent; Walker, Deandra L; Markham, Neil E; Balasubramaniam, Vivek; Abman, Steven H

    2010-12-01

    To determine the separate and interactive effects of fetal inflammation and neonatal hyperoxia on the developing lung, we hypothesized that: 1) antenatal endotoxin (ETX) causes sustained abnormalities of infant lung structure; and 2) postnatal hyperoxia augments the adverse effects of antenatal ETX on infant lung growth. Escherichia coli ETX or saline (SA) was injected into amniotic sacs in pregnant Sprague-Dawley rats at 20 days of gestation. Pups were delivered 2 days later and raised in room air (RA) or moderate hyperoxia (O₂, 80% O₂ at Denver's altitude, ∼65% O₂ at sea level) from birth through 14 days of age. Heart and lung tissues were harvested for measurements. Intra-amniotic ETX caused right ventricular hypertrophy (RVH) and decreased lung vascular endothelial growth factor (VEGF) and VEGF receptor-2 (VEGFR-2) protein contents at birth. In ETX-exposed rats (ETX-RA), alveolarization and vessel density were decreased, pulmonary vascular wall thickness percentage was increased, and RVH was persistent throughout the study period compared with controls (SA-RA). After antenatal ETX, moderate hyperoxia increased lung VEGF and VEGFR-2 protein contents in ETX-O₂ rats and improved their alveolar and vascular structure and RVH compared with ETX-RA rats. In contrast, severe hyperoxia (≥95% O₂ at Denver's altitude) further reduced lung vessel density after intra-amniotic ETX exposure. We conclude that intra-amniotic ETX induces fetal pulmonary hypertension and causes persistent abnormalities of lung structure with sustained pulmonary hypertension in infant rats. Moreover, moderate postnatal hyperoxia after antenatal ETX restores lung growth and prevents pulmonary hypertension during infancy.

  16. Evaluation of Amniotic-Derived Membrane Biomaterial as an Adjunct for Repair of Critical Sized Bone Defects

    OpenAIRE

    Starecki, Mikael; Schwartz, John A.; Grande, Daniel A.

    2014-01-01

    Introduction. Autogenous bone graft is the gold standard in reconstruction of bone defects. The use of autogenous bone graft is problematic because of limited bone as well as donor site morbidity. This study evaluates a novel biomaterial as an alternative to autogenous bone graft. The biomaterial is amniotic membrane, rich in growth factors. Methods. Twenty-one adult male Sprague-Dawley rats were implanted with biomaterial using the rat critical size femoral gap model. After creation of the c...

  17. Reliable prenatal diagnosis of Canavan disease by measuring N-acetylaspartate in amniotic fluid using liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Al-Dirbashi, Osama Y; Kurdi, Wesam; Imtiaz, Faiqa; Ahmad, Asmahan M; Al-Sayed, Moeenaldeen; Tulbah, Maha; Al-Nemer, Maha; Rashed, Mohamed S

    2009-05-01

    Prenatal diagnosis of Canavan disease by measuring N-acetylaspartic acid (NAA) in amniotic fluid is reliable and preferred over aspartoacylase enzyme assay especially in populations with unknown mutations. Typically based on GC-MS, existing methods are time-consuming and laborious. We developed a novel LC-MS/MS method for determination of NAA in amniotic fluid with minimal sample preparation. NAA and d(3)-NAA were detected by negative-ion electrospray ionization-MS/MS. Quantification was achieved by standard addition using six 0.1 mL portions of each specimen enriched with increasing NAA amounts (0, 0.05, 0.1, 0.2, 0.3, and 0.4 microg) and endogenous NAA was calculated by extrapolation. Injection-to-injection time was 2 min whereas the turn around time from sample receipt was about 1 h. Intraday (n = 10) and interday (n = 10) variations were less than 9.4%. The reference range determined using gestation-matched controls (n = 12) of 1.1-2.7 micromol/L is in agreement with the literature. Specimens from at-risk pregnancies with established diagnosis (n = 4) were successfully analyzed. We developed a new method that enables reliable, sensitive, and selective determination of NAA in a small volume of amniotic fluid for the prenatal diagnosis of Canavan disease. The simple sample preparation adopted in this work precluded the necessity for extraction and derivatization. (c) 2009 John Wiley & Sons, Ltd.

  18. Combined Excimer Laser Photoablation and Amniotic Membrane Overlay for Relief of Symptomatic Discomfort in Gelatinous Drop-like Corneal Dystrophy.

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    Alex, Anne F; Eter, Nicole; Uhlig, Constantin E

    2015-10-01

    To describe the efficacy of combined excimer laser photoablation and amniotic overlay membrane in the relief of symptomatic discomfort in a 17-year-old patient who had gelatinous drop-like corneal dystrophy. The best-corrected visual acuity (BCVA) was measured with Snellen letters. Slit-lamp examination of the ocular surface and anterior chamber was performed at baseline. Results were photodocumented. Excimer laser photoablation was performed and subsequently 2 amniotic membranes were transconjunctivally fixated with 10.0 nylon sutures. Investigations and documentation were performed at baseline, every 2 months in the first year, and then every 6 months. The duration of follow-up was 22 months. At baseline, the BCVA was 20/70 in the right eye and 20/200 in the left eye. The patient reported distinct photophobia. Slit-lamp examination was difficult because of blepharospasm. Although gelatinous drops developed again and the BCVA decreased to 2/200, the patient reported significant relief after both microsurgical treatments and remained comfortable at 20 and 22 months. Excimer laser photocoagulation combined with amniotic membrane overlay does not stop the development of gelatinous drop-like corneal dystrophy but may improve subjective comfort. Such treatment does not hinder subsequent lamellar or penetrating grafts and is helpful in providing the necessary time for preparation of matched keratoplasties.

  19. Microstructure and glycosaminoglycan ratio of canine cornea after reconstructive transplantation with glycerin-preserved porcine amniotic membranes.

    Science.gov (United States)

    Tsuzuki, Keiko; Yamashita, Kazuto; Izumisawa, Yasuharu; Kotani, Tadao

    2008-01-01

    Although amniotic membranes of canine, feline, and equine species have some advantages as corneal transplantation material in many canine ocular diseases, their softness, thinness, and low availability can pose problems. As an alternative, the more abundant porcine amniotic membranes may be used. This paper describes the use of glycerin-preserved porcine amniotic membranes in corneal transplantation in eight normal dogs. A 0.4-mm deep recipient bed in the axial cornea of the OS of all dogs was created using an 8-mm Barron radial vacuum trephine. The recipient bed was then filled with amnion, and the entire cornea was covered with another piece of the glycerin-preserved membrane. The ocular signs evaluated were corneal opacity and corneal vascularization. The dogs were euthanized on days 5, 10, 20, or 40 after surgery, and samples were collected to evaluate corneal thickness, parenchymal cell number, mean collagen fibril diameter, collagen fibril content and the glycosaminoglycan (GAG) ratio. Corneal opacity was observed immediately after surgery. Restoration of corneal transparency, regression of corneal vascularization, and visualization of the pupil and iris were noted on day 40. The clinical observations were supported histologically by regained corneal thickness, parenchymal cell number, mean collagen fibril diameter, collagen fibril content, and GAG ratio, suggesting that this technique may be a novel method for the treatment of ocular surface disorders.

  20. Allogeneic amniotic membrane-derived mesenchymal stromal cell transplantation in a porcine model of chronic myocardial ischemia

    Science.gov (United States)

    Kimura, M; Toyoda, M; Gojo, S; Itakura, Y; Kami, D; Miyoshi, S; Kyo, S; Ono, M; Umezawa, A

    2012-01-01

    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. PMID:24693195

  1. Can quantity of amniotic fluid reliably predict postnatal renal function in boys with posterior urethral valves: a decision curve analysis.

    Science.gov (United States)

    Harper, Luke; Waubant, Alice; Vignes, Julien; Amat, Sara; Dobremez, Eric; Lefevre, Yan; Ferdynus, Cyril

    2017-09-01

    Prenatal management of male fetuses with suspected posterior urethral valves depends on reliable markers for postnatal long-term renal function. Whether ultrasound parameters, including the presence or absence of oligohydramnios, are reliable remains the subject of debate. We decided to evaluate the reliability of quantity of amniotic fluid to predict postnatal renal function using decision curve analysis (DCA), a method for evaluating the clinical utility of a diagnostic test. We analyzed retrospectively 51 male fetuses born with prenatally suspected posterior urethral valves between 2009 and 2012. We studied the relationship between quantity of amniotic fluid on prenatal ultrasound and the nadir creatinine during the first year of life as a proxy of postnatal renal function using DCA. Twelve fetuses presented with prenatal oligohydramnios. Thirty-one children had a normal nadir creatinine, of which one had prenatal oligohydramnios (3.2%). Thirteen had a nadir creatinine between 35 and 75 μmol/L, of which four had prenatal oligohydramnios (30.8%). Seven had a nadir creatinine >75 μmol/L, all of them had prenatal oligohydramnios. In this retrospective study, DCA confirms the relationship between prenatal quantity of amniotic fluid volume and postnatal renal function. © 2017 John Wiley & Sons, Ltd. © 2017 John Wiley & Sons, Ltd.

  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. Influence of maternal age, gestational age and fetal gender on expression of immune mediators in amniotic fluid.

    Science.gov (United States)

    Weissenbacher, Tobias; Laubender, Rüdiger P; Witkin, Steven S; Gingelmaier, Andrea; Schiessl, Barbara; Kainer, Franziskus; Friese, Klaus; Jeschke, Udo; Dian, Darius; Karl, Katrin

    2012-07-24

    Variations in cytokine and immune mediator expression patterns in amniotic fluid due to gestational age, maternal age and fetal gender were investigated. Amniotic fluid samples were obtained from 192 women, 82 with a mid-trimester amniocentesis (median gestational age 17 weeks) and 110 with a caesarean section not in labor (median gestational age 39 weeks). Amniotic fluid was screened by commercial ELISAs for the TH1/TH2/TH17 cytokines and immune mediators IL-1 beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-15, IL-17, TNF alpha, GRO-alpha, MIP1alpha, MIP1beta, Histone, and IP10. Analysis was by Bonferroni correction for multiple comparisons. None of the 15 examined cytokines revealed any differences in expression patterns regarding fetal gender. Significant differences were found in IL-4, IL-10, IL-12, TNF- alpha, GRO-alpha and MIP1-beta with respect to gestational age and in GRO-alpha regarding maternal age. Cytokines utilized as biomarkers in the diagnosis of intrauterine infections are not influenced in their expression pattern by fetal gender but may vary with respect to maternal age and gestational age.

  4. Evaluation of Amniotic-Derived Membrane Biomaterial as an Adjunct for Repair of Critical Sized Bone Defects

    Directory of Open Access Journals (Sweden)

    Mikael Starecki

    2014-01-01

    Full Text Available Introduction. Autogenous bone graft is the gold standard in reconstruction of bone defects. The use of autogenous bone graft is problematic because of limited bone as well as donor site morbidity. This study evaluates a novel biomaterial as an alternative to autogenous bone graft. The biomaterial is amniotic membrane, rich in growth factors. Methods. Twenty-one adult male Sprague-Dawley rats were implanted with biomaterial using the rat critical size femoral gap model. After creation of the critical size femoral gap animals were randomized to one of the following groups: Group 1 (control: gap left empty and received no treatment; Group 2 (experimental: the gap was filled with commercially available bone graft; Group 3 (experimental: the gap was filled with bone graft plus NuCel amniotic tissue preparation. Results. The experimental groups demonstrated new bone formation compared to controls. The results were evident on radiographs and histology. Histology showed Group 1 controls to have 11.1% new bone formation, 37.8% for Group 2, and 49.2% for Group 3. These results were statistically significant. Conclusions. The study demonstrates that amniotic membrane products have potential to provide bridging of bone defects. Filling bone defects without harvesting autogenous bone would provide a significant improvement in patient care.

  5. miR-26a Mediates Adipogenesis of Amniotic Fluid Mesenchymal Stem/Stromal Cells via PTEN, Cyclin E1, and CDK6.

    Science.gov (United States)

    Trohatou, Ourania; Zagoura, Dimitra; Orfanos, Nikos K; Pappa, Kalliopi I; Marinos, Evangelos; Anagnou, Nicholas P; Roubelakis, Maria G

    2017-04-01

    Recent findings indicate that microRNAs (miRNAs) are critical for the regulatory network of adipogenesis in human mesenchymal stem/stromal cells (MSCs). Fetal MSCs derived from amniotic fluid (AF-MSCs) represent a population of multipotent stem cells characterized by a wide range of differentiation properties that can be applied in cell-based therapies. In this study, miRNA microarray analysis was performed to assess miRNA expression in terminal differentiated AF-MSCs into adipocyte-like cells (AL cells). MiR-26a was identified in high expression levels in AL cells indicating a critical role in the process of adipogenesis. Overexpression of miR-26a in AF-MSCs led to significant induction of their adipogenic differentiation properties that were altered after miR-26a inhibition. We have demonstrated that miR-26a regulates adipogenesis through direct inhibition of PTEN, which in turn promotes activation of Akt pathway. Also, miR-26a modulates cell cycle during adipogenesis by interacting with Cyclin E1 and CDK6. These results point to the regulatory role of miR-26a and its target genes PTEN, Cyclin E1, and CDK6 in adipogenic differentiation of AF-MSCs, providing a basis for understanding the mechanisms of fat cell development and obesity.

  6. Prospective, randomized, blinded, comparative study of injectable micronized dehydrated amniotic/chorionic membrane allograft for plantar fasciitis--a feasibility study.

    Science.gov (United States)

    Zelen, Charles M; Poka, Attila; Andrews, James

    2013-10-01

    Specialized treatment of plantar fasciitis that can reduce inflammation and promote healing may be a possible alternative prior to surgical intervention. We report the results of a randomized clinical trial examining the efficacy of micronized dehydrated human amniotic/chorionic membrane (mDHACM) injection as a treatment for chronic refractory plantar fasciitis. An institutional review board-approved, prospective, randomized, single-center clinical trial was performed. Forty-five patients were randomized to receive injection of 2 cc 0.5% Marcaine plain, then either 1.25 cc saline (controls), 0.5 cc mDHACM, or 1.25 cc mDHACM. Follow-up visits occurred over 8 weeks to measure function, pain, and functional health and well-being. Significant improvement in plantar fasciitis symptoms was observed in patients receiving 0.5 cc or 1.25 cc mDHACM versus controls within 1 week of treatment and throughout the study period. At 1 week, American Orthopaedic Foot and Ankle Society (AOFAS) Hindfoot scores increased by a mean of 2.2 ± 17.4 points for controls versus 38.7 ± 11.4 points for those receiving 0.5 cc mDHACM (P plantar fasciitis, mDHACM is a viable treatment option. Larger studies are needed to confirm our findings. Level I, prospective randomized study.

  7. The use of amniotic membrane in the repair of duodenal wounds in Wistar rats Uso da membrana amniótica no reparo de feridas duodenais em ratos Wistar

    Directory of Open Access Journals (Sweden)

    Luciano Rodrigues Schimidt

    2010-02-01

    Full Text Available Purpose: In the search of a new material to repair duodenal wounds, a trial was conducted to assess the behavior of human amniotic membrane in the repair of the duodenal wall in rats. METHODS: Fifty Wistar rats weighing between 250 and 350g, male, were submitted to duodenotomy and randomly distributed into two groups. Group A (n=8 had no treatment and was used as the control group. In Group B (n=42 the duodenal wound was treated with a patch of human amniotic membrane. RESULTS: All animals in Group A died. In Group B no changes were observed with regards to death or the formation of duodenal fistula. All animals presented peritoneal adherences in the region on the duodenal wall repair and intestinal obstruction was observed in two animals. Healing of the duodenal wall in the region of the patch took place progressively as the post-operatory period increased, with regeneration of the mucosa and of the smooth muscle layer. CONCLUSION: From the clinical standpoint, the amniotic membrane proved to be a biological tissue which served as a temporary seal and allowed the wound to heal by second-intention, with re-establishment of the duodenal wall structure.OBJETIVO: Na busca de um novo material para o reparo dos ferimentos duodenais, foi efetuado um estudo para avaliar o comportamento da membrana amniótica humana no reparo da parede duodenal em ratos. MÉTODOS: Foram utilizados 50 ratos Wistar, com peso entre 250 e 350g, machos, distribuídos, aleatoriamente, em dois grupos. Grupo A (n=8, submetido à duodenotomia sem tratamento, utilizados como controle. Grupo B (n=42, submetido a um remendo de membrana amniótica humana para tratamento de ferimento duodenal provocado. RESULTADOS: Todos os animais do grupo A foram a óbito. No grupo B não foram observadas alterações quanto a óbito ou formação de fístula duodenal. Observaram-se em todos os animais aderências peritoneais à região do reparo da parede do duodeno e obstrução intestinal em

  8. [Amniotic membrane transplantation and high-dose systemic cyclosporin A (Sandimmun optoral) for Mooren's ulcer].

    Science.gov (United States)

    Spelsberg, H; Sundmacher, R

    2007-02-01

    Mooren's ulcer is a rare, severe corneal autoimmune inflammation leading to blindness if treated insufficiently. High-dose systemic cyclosporin A (Sandimmun optoral) was shown to markedly reduce inflammation and stop corneal destruction. We report on three cases in which this immunosuppressive regimen required additional AM transplantation for complete healing. One 37-year-old male (M) and two 49- (F1) and 58-year-old females (F2) presented with unilateral Mooren's ulcer in different stages of the disease, which deteriorated despite high-dose systemic cyclosporin A (Sandimmun optoral M, F1) or treatment with topical cyclosporin A 2 % (F2). After surgical removal of all grossly affected corneal stroma, amniotic membrane was made to cover the entire cornea and fixed with episcleral sutures in two patients (M, F2). In one patient (F1) a deep marginal ulcer was covered with a fitted AM glap. All patients were treated postoperatively up to 6 months with high-dose systemic cyclosporin A (Sandimmun optoral). Blood trough levels aimed at 150 to 200 ng/mL. Topical cyclosporin A was administered in addition in two patients (M, F2) postoperatively for at least 6 months. Due to incompatibility, one patient (F1) was treated with topical steroids instead. Follow-up time was 42 months (M), 50 months (F1) and 54 months (F2). All three eyes exhibited clinical healing with stable corneal surfaces thereafter. Depending on the stage of Mooren's ulcer at the time of surgery, visual acuity remained at hand motions in one patient (M) and recovered to 1.0 and 0.6, respectively, in two patients (F1, F2). Due to its anti-inflammatory potential, coverage by AM seems to trigger a therapeutic turnaround in cases of Mooren's ulcer which do not heal with intensive immunosuppressive regimens alone. In order to maintain or restore as much visual function as possible, additional amniotic membrane surgery should be performed early enough in the course of the disease.

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

    Science.gov (United States)

    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

  10. Should fetal scalp blood sampling be performed in the case of meconium-stained amniotic fluid?

    Science.gov (United States)

    Boujenah, J; Oliveira, J; De La Hosseraye, C; Benbara, A; Tigaizin, A; Bricou, A; Carbillon, L

    2016-12-01

    To investigate the effect of using fetal scalp blood sampling on the risk of neonatal respiratory distress syndrome (NRDS) with meconium-stained amniotic fluid (MSAF). Prospective data collection with regard to MSAF during labor for low-risk term cephalic singleton live birth from 2012 to 2014. Maternal, obstetric and neonatal data were compared according to the occurrence of respiratory distress syndrome (RDS group) or not (no RDS group). Of 515 newborns born through MSAF, 46 experienced RDS and from them 10 experienced meconium aspiration syndrome. No difference was observed according to maternal characteristic, abnormal fetal heart rate tracing pattern irrespective of its category and cesarean rate. Apgar at one minute was lower in the group RDS (7.6 versus 8.5, p scalp lactate sampling during the labor (71.1% versus 55.1%, p scalp blood assessment (p scalp blood assessment and MSAF diagnosed during the first stage of labor (after spontaneous rupture of membranes or at amniotomy) was found. In case of MSAF, fetal scalp blood sampling did not reduce the risk of RDS.

  11. A modified symblepharon ring for sutureless amniotic membrane patch to treat acute ocular surface burns.

    Science.gov (United States)

    Liang, Xuanwei; Liu, Zhiping; Lin, Ying; Li, Naiyang; Huang, Minghai; Wang, Zhichong

    2012-01-01

    The objective of this study is to evaluate a sutureless technique by using a modified symblepharon ring to fix an amniotic membrane (AM) patch on the ocular surface to treat acute ocular burns. Seventy-five patients with acute ocular burns of total 75 eyes graded III to VI were enrolled in this study. They were randomly divided into two groups. Thirty-nine eyes received the sutureless AM patch with a modified symblepharon ring, and the other 36 eyes underwent the conventional sutured AM patch as control. The time and the rate of epithelialization, corneal neovascularization, and complications were recorded. Both the operation time and the time to epithelial closure in the sutureless group were much shorter than that in the suture group (P < .01). The rate of reepithelialization in the sutureless group was higher than in the suture group (P < .05). The rate of the vascularization and symblepharon were lower in the sutureless group than in the suture group (P < .05). The conjunctival sac contraction occurred only in the eyes with grade V and VI in the sutureless group and was later than in the suture group (P < .05). This modified method is simple, minimally invasive, free from trauma, and more effective compared with controls.

  12. Sheep CD34+ amniotic fluid cells have hematopoietic potential and engraft after autologous in utero transplantation.

    Science.gov (United States)

    Shaw, S W Steven; Blundell, Michael P; Pipino, Caterina; Shangaris, Panicos; Maghsoudlou, Panagiotis; Ramachandra, Durrgah L; Georgiades, Fanos; Boyd, Michael; Thrasher, Adrian J; Porada, Christopher D; Almeida-Porada, Graça; Cheng, Po-Jen; David, Anna L; de Coppi, Paolo

    2015-01-01

    Unmatched allogeneic in utero stem cell transplantation (IUSCT) produces poor engraftment unless the fetus has congenital immunodeficiency, probably because of maternal and fetal immune responses to injected cells. We studied the functional hematopoietic potential of transduced green fluorescent protein (GFP+) sheep amniotic fluid (AF) stem cells, before and after autologous IUSCT. CD34+ cells were selected from first trimester sheep AF, transduced overnight, and injected intravenously into NOD-SCID-gamma (NSG) mice. At 3 months, primary recipient bone marrow (BM) was injected into secondary NSG recipients. GFP+ cells were detected in the hematopoietic organs and peripheral blood of primary and secondary recipients at 3 months. Autologous IUSCT (transduced GFP+CD34+AF) was performed in fetal sheep. Six months postnatally, lamb BM was injected into secondary NSG recipients. GFP+ cells were detected in the peripheral blood of primary and secondary recipients. This confirms the hematopoietic potential of AF stem cells supporting the concept of autologous IUSCT to treat congenital hematopoietic disease. © 2014 AlphaMed Press.

  13. Meconium-stained amniotic fluid--risk factors and immediate perinatal outcomes among SGA infants.

    Science.gov (United States)

    Pariente, Gali; Peles, C; Perri, Zvi H; Baumfeld, Yael; Mastrolia, Salvatore Andrea; Koifman, Arie; Weintraub, Adi Y; Hershkovitz, Reli

    2015-06-01

    To detect factors that are associated with meconium-stained amniotic fluid (MSAF) among deliveries of small for gestational age (SGA) neonates and to identify perinatal outcomes of deliveries of SGA infants complicated with MSAF. A population-based study comparing deliveries of SGA neonates with and without MSAF was conducted. Deliveries occurred during the years 1988-2007 at the Soroka University Medical Center. Risk factors for MSAF among SGA infants were evaluated. Incidence of adverse pregnancy outcomes were compared between deliveries of SGA neonates with and without MSAF. During the study period 9583 deliveries were of SGA neonates. Of these, 16.6% (n = 1597) were complicated with MSAF. Among SGA neonates, older maternal age, multiparty, lack of prenatal care and weight were significantly associated with MSAF. Having delivered an SGA infant with MSAF was associated with decreased rates of induction of labor and increased rates of labor dystocia, delivery by cesarean section and fetal distress. Using multivariable regression models, having delivered an SGA infant with MSAF was independently associated with fetal distress. Among SGA neonates, deliveries complicated with MSAF are associated with additional adverse pregnancy outcomes.

  14. Randomized Trial Comparing Amniotic Membrane Transplantation with Lamellar Corneal Graft for the Treatment of Corneal Thinning.

    Science.gov (United States)

    de Farias, Charles C; Allemann, Norma; Gomes, José Á P

    2016-04-01

    There are few studies comparing different surgical procedures for the treatment of corneal thinning. Lamellar corneal transplantation (LCT) has been reported to be efficient, but its results can be jeopardized by allograft rejection, opacification, or high astigmatism. Amniotic membrane transplantation (AMT) has been considered a good alternative, but it is not as resistant as LCT and the tissue can be reabsorbed after surgery. A prospective, randomized, interventional, and comparative study of consecutive patients with corneal thinning over 6 months was performed. Ophthalmological examination was performed before transplant surgery and then repeated 1, 7, 15, 30, 90, and 180 days after surgery and ultrasound biomicroscopy was performed before and then 30, 90, and 180 days after surgery to assess corneal thinning. Herpes simplex infection was the main cause of corneal thinning (9 eyes), followed by surgery (cataract, glaucoma, 5 cases), rheumatoid arthritis (1), chemical burn (1), perforating trauma (1), previous band keratopathy treatment (1), and Stevens-Johnson syndrome (1). Although all patients showed significant increase in final thickness in the area of thinning, it was higher in those submitted to LCT at 180 days postoperatively. Regardless of the surgical technique, all patients showed epithelialization. Patients undergoing AMT showed an 89% decrease in neovascularization. Final corrected distance visual acuity was better in patients submitted to AMT. LCT proved to be the best option for treating corneal thinning. AMT represents an alternative that allows good visual recovery but does not restore corneal thickness as efficiently as LCT.

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

    Science.gov (United States)

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

    1999-01-01

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

  16. Clinical observation of cryotherapy and amniotic membrane transplantation in the management of Fusarium corneal ulcer

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    Qiong Xie

    2014-10-01

    Full Text Available AIM:To examine the efficacy and safety of cryotherapy in the management of Fusarium corneal ulcer. METHODS: Retrospective contract analysis of the clinical data of 41 patients(41 eyeswho infected with Fusarium corneal ulcer. All of them underwent focal lesion keratectomy combined with amniotic membrane transplantation between January 2010 and May 2013. The cryotherapy treatment group of 22 cases(22 eyes, non-cryotherapy group of 19 cases(19 eyes. All the cases were followed up for 3~12mo. We analyzed the healing of corneal ulcer, corneal neovascularization, postoperative visual acuity, complications and ulcer recurrences. RESULTS: There were no significant difference for the two groups at the cure rate and the recovery rate, the corneal neovascularization and postoperative visual acuity(P>0.05. The total effective rate and corneal healing time in cryotherapy group was significantly higher than that in non-cryotherapy group(PPCONCLUSION: Cryotherapy is an effective and safe method for the treatment of Fusarium corneal ulcer.

  17. The experience of women in hospital rest with preterm premature rupture of de amniotic membranes

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    María del Mar Fernández Míguez

    2012-07-01

    Full Text Available Hospital bed rest in a preterm premature rupture of the amniotic membranes involves a crisis situation for women with very high emotional effects.To understand the needs felt of the gestantes with pregnancies of high risk on the part of the professionals, it will improve the quality of the attention and it can help to diminish the stress levels at this vulnerable period and to confronting the maternity.Aim: Investigate women´s experience when they are resting in hospital in cases de preterm premature rupture of membranes.Methodology: Phenomenological study, It has taken as a population of study to women hospitalized in the plant of obstetrics of high risk from Gregorio Marañón Hospital, with preterm premature rupture of membranes between 24 and 31 weeks of gestation.Data collection: Was be carried out by means of the individual interviews in hospitable environment; observation and withdrawal of notes during the accomplishment of this one.

  18. Bilateral Mooren’s ulcer – Customised corneal graft with additional amniotic membrane graft

    Science.gov (United States)

    Bhandari, Vipul; Siddharthan, K.S.

    2015-01-01

    Mooren’s ulcer (MU) is a rare and painful peripheral corneal ulceration which occurs in the absence of any associated scleritis, and any detectable systemic disease. A 60-year-old male patient was referred to us with bilateral peripheral corneal ulceration. Best corrected visual acuity (BCVA) in both eyes was counting finger at one metre. The right eye showed a 180° thinning with perforation at 8 o’clock position. The left eye showed a 360° thinning with central contact lens type cornea. After complete blood analysis we started the patient on cyclophosphamide orally along with high doses of oral steroids. A crescentic excision of the thinned cornea and crescentic customised corneal graft with additional amniotic membrane graft (AMG) was done first for the right eye and a 360° peripheral lamellar corneal graft with additional AMG for the left eye. The BCVA of RE was 1/60 improving to 6/36 with plus ten aphakic glasses and LE was 3/60. Hand fashioned full thickness crescentic customised corneal graft with additional AMG and a peripheral 360° lamellar corneal graft with additional AMG in these cases are a novel approach to Mooren’s ulcer with gratifying results. PMID:26155087

  19. Bilateral Mooren's ulcer - Customised corneal graft with additional amniotic membrane graft.

    Science.gov (United States)

    Bhandari, Vipul; Siddharthan, K S

    2015-01-01

    Mooren's ulcer (MU) is a rare and painful peripheral corneal ulceration which occurs in the absence of any associated scleritis, and any detectable systemic disease. A 60-year-old male patient was referred to us with bilateral peripheral corneal ulceration. Best corrected visual acuity (BCVA) in both eyes was counting finger at one metre. The right eye showed a 180° thinning with perforation at 8 o'clock position. The left eye showed a 360° thinning with central contact lens type cornea. After complete blood analysis we started the patient on cyclophosphamide orally along with high doses of oral steroids. A crescentic excision of the thinned cornea and crescentic customised corneal graft with additional amniotic membrane graft (AMG) was done first for the right eye and a 360° peripheral lamellar corneal graft with additional AMG for the left eye. The BCVA of RE was 1/60 improving to 6/36 with plus ten aphakic glasses and LE was 3/60. Hand fashioned full thickness crescentic customised corneal graft with additional AMG and a peripheral 360° lamellar corneal graft with additional AMG in these cases are a novel approach to Mooren's ulcer with gratifying results.

  20. Amniotic epithelial cells: a new tool to combat aging and age-related diseases?

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    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.

  1. Surgical resection and amniotic membrane transplantation for treatment of refractory giant papillae in vernal keratoconjunctivitis.

    Science.gov (United States)

    Guo, Ping; Kheirkhah, Ahmad; Zhou, Wei-wei; Qin, Lei; Shen, Xiao-li

    2013-06-01

    The aim of this study was to evaluate the outcome of surgical resection and amniotic membrane transplantation (AMT) for treatment of refractory symptomatic giant papillae in vernal keratoconjunctivitis (VKC). This is a retrospective study of 13 eyes of 9 patients with refractory giant papillae associated with corneal shield ulcer and/or punctate epithelial erosions who underwent surgical resection of the papillae combined with AMT to cover the tarsal conjunctival defect. During 14.2 ± 4.2 months of postoperative follow-up, smooth tarsal conjunctival surface was achieved in all cases, with no recurrence of the giant papillae in any eye. Corneal shield ulcers and punctate epithelial erosions healed within 2 weeks after surgery and did not recur during the follow-up. Best-corrected visual acuity improved from 0.26 ± 0.21 logarithm of the minimum angle of resolution preoperatively to 0.02 ± 0.04 logarithm of the minimum angle of resolution postoperatively (P = 0.01). Three patients experienced recurrence of VKC symptoms, but without giant papillae, which could be well controlled by topical medications. Surgical resection combined with AMT is an effective procedure for treatment of refractory giant papillae in patients with VKC.

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

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    Hamid Gourabi

    2008-01-01

    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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  6. Phylogenomics of nonavian reptiles and the structure of the ancestral amniote genome

    Science.gov (United States)

    Shedlock, Andrew M.; Botka, Christopher W.; Zhao, Shaying; Shetty, Jyoti; Zhang, Tingting; Liu, Jun S.; Deschavanne, Patrick J.; Edwards, Scott V.

    2007-01-01

    We report results of a megabase-scale phylogenomic analysis of the Reptilia, the sister group of mammals. Large-scale end-sequence scanning of genomic clones of a turtle, alligator, and lizard reveals diverse, mammal-like landscapes of retroelements and simple sequence repeats (SSRs) not found in the chicken. Several global genomic traits, including distinctive phylogenetic lineages of CR1-like long interspersed elements (LINEs) and a paucity of A-T rich SSRs, characterize turtles and archosaur genomes, whereas higher frequencies of tandem repeats and a lower global GC content reveal mammal-like features in Anolis. Nonavian reptile genomes also possess a high frequency of diverse and novel 50-bp unit tandem duplications not found in chicken or mammals. The frequency distributions of ≈65,000 8-mer oligonucleotides suggest that rates of DNA-word frequency change are an order of magnitude slower in reptiles than in mammals. These results suggest a diverse array of interspersed and SSRs in the common ancestor of amniotes and a genomic conservatism and gradual loss of retroelements in reptiles that culminated in the minimalist chicken genome. PMID:17307883

  7. Sealing of Corneal Lacerations Using Photoactivated Rose Bengal Dye and Amniotic Membrane.

    Science.gov (United States)

    Soeken, Timothy A; Zhu, Hong; DeMartelaere, Sheri; Davies, Brett W; Kim, Mirang; Wang, Heuy-Ching; Aden, James; Grimm, Rose; Alt, Clemens; Kochevar, Irene E; Johnson, Anthony J

    2018-02-01

    Watertight closure of perforating corneoscleral lacerations is necessary to prevent epithelial ingrowth, infection, and potential loss of the eye. Complex lacerations can be difficult to treat, and repair with sutures alone is often inadequate. In this study, we evaluated a potentially sutureless technology for sealing complex corneal and scleral lacerations that bonds the amniotic membrane (AM) to the wound using only green light and rose bengal dye. The AM was impregnated with rose bengal and then sealed over lacerations using green light to bond the AM to the deepithelialized corneal surface. This process was compared with suture repair of 3 laceration configurations in New Zealand White rabbits in 3 arms of the study. A fourth study arm assessed the side effect profile including viability of cells in the iris, damage to the blood-retinal barrier, retinal photoreceptors, retinal pigment epithelium, and choriocapillaris in Dutch Belted rabbits. Analyses of the first 3 arms revealed a clinically insignificant increase in polymorphonuclear inflammation. In the fourth arm, iris cells appeared unaffected and no evidence of breakdown of the blood-retinal barrier was detected. The retina from green light laser-treated eyes showed normal retinal pigment epithelium, intact outer segments, and normal outer nuclear layer thickness. The results of these studies established that a light-activated method to cross-link AM to the cornea can be used for sealing complex penetrating wounds in the cornea and sclera with minimal inflammation or secondary effects.

  8. 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.

  9. Hyperdry amniotic membrane as a suitable biological dressing material for raw wounds in the oral cavity

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    Makoto Noguchi

    2016-06-01

    Full Text Available Raw wounds in the oral cavity are prone not only to infection but also contraction by scaring and often need a proper dressing to prevent these complications. Autografts using free mucosal and split-skin grafts, which seem biologically ideal, have been used to cover raw wounds in the oral cavity. Those grafts, however, require a separate surgical procedure at donor sites and often cause morbidity associated with delayed healing of the donor site. The amnion has been considered a suitable tissue for allografts, based on its low immunogenicity. It also possesses anti-inflammatory, would –protecting, and scar-reducing properties. Preserved amnions have been used for decades in various clinical fields.  However, there have been some problems in the storage and sterilization of the material. To resolve these problems, we developed hyperdry amniotic membrane (HAM, which can be stored at room temperature for a long period. In my lecture, I will share our clinical experiences of applying HAM into oral surgery, including results of experimental studies on would healing of the oral cavity.

  10. 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

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    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.

  11. Comparison of Extra-Amniotic Normal Saline Infusion plus Hydrocortisone versus Prostaglandin E2 Suppository for Pregnancy Termination

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    Farahnaz Keshavarzi

    2012-08-01

    Full Text Available Background and Aim: Pregnancy termination before onset of labor is one of the midwifery problems. The aim of the present study was to compare extra-amniotic normal saline infusion plus hydrocortisone effect versus prostaglandin E2 suppository for cervical ripening and second- trimester pregnancy termination.   Materials and Methods: This randomized clinical-trial study was performed on 50 pregnant women with gestation age of 14 to 28 weeks and Bishops Score ≤ 2 who required pregnancy termination for fetal indications. Half of the cases received extra-amniotic normal saline infusion plus 100 mg hydrocortisone (EASI+ H and the other half received two doses of 3mg prostaglandin E2 suppository every 4 hours. Six hours later, induction of labor was started by means of a high dose of Oxytocin according to of Alabama University Protocol for Mid-trimester Abortion. The obtained data was analyzed by using statistical softwares Chi-square, Fisher and T-test.   Results: There were no differences between the two groups regarding maternal age, parity, gestational age, Primary Bishops Score and indication of pregnancy termination. In the EASI+H group, the mean interval between initiation of labor induction and fetal expulsion was 23.04 ± 4.47 hours and in PGE2 group was 28.65 ± 2.87 hours (P=0.001. The success rate in the EASI+H group was 100% and in PGE2 group 80 % (P =0.04. Complications such as fever, nausea, vomiting, diarrhea, increase of blood pressure and need to curettage in PGE2 group were statistically and significantly more abundant.   Conclusion: Extra-amniotic normal saline infusion plus hydrocortisone is an effective and safe method that is suggested for cervical ripening and second- trimester pregnancy termination.

  12. A Retrospective Study of Cytogenetic Results From Amniotic Fluid in 5328 Fetuses With Abnormal Obstetric Sonographic Findings.

    Science.gov (United States)

    Zhang, Shuo; Lei, Caixia; Wu, Junping; Sun, Haiyan; Yang, Yuezhou; Zhang, Yueping; Sun, Xiaoxi

    2017-09-01

    The purpose of this study was to evaluate the diagnostic utility of karyotype analysis of amniotic fluid for fetuses with abnormal sonographic findings and to determine the detection rates of abnormal karyotypes. We conducted a retrospective study of 5328 fetuses with abnormal sonographic findings in the first or second trimester enrolled from October 1998 and September 2015. Cytogenetic results from amniotic fluid were obtained in all of these pregnancies. Sonographic abnormalities were stratified according to anatomic system involvement. A total of 238 abnormal karyotypes were encountered in the 5328 fetuses (4.5%). The highest rate of chromosomal anomalies was in fetuses with structural abnormalities in multiple organ systems (25.7%), followed by an abnormal amniotic fluid volume (7.9%), structural abnormalities in a single system (7.3%), multiple nonstructural anomalies (7.2%), isolated placental abnormalities (7.1%), and isolated soft markers for aneuploidy (2.4%; P abnormalities in a single system, gastrointestinal and neck/body fluids had particularly high detection rates (26.1% and 26.2%, respectively). A detailed analysis suggest