archives@tulane.edu / Amniotic fluid (AF) and amniotic membrane (AM) have been identified as potential new sources of stem cells. While numerous studies have confirmed the presence of mesenchymal stem cells (MSCs) in AM cells, limited research exists regarding full-term AF cells. The goal of this research was to assess the ability to process full-term AF and AM and characterize cell populations isolated from these sources. Specifically, their MSC capability was explored by studying their stemness marker expression, adipogenic and osteogenic differentiation potential, and ability to proliferate and form colonies. AF and AM samples were obtained from consenting donors who underwent elected Cesarean sections. Flow cytometry analysis showed positive expression of MSC markers in AF and AM cells. AF and AM cells exhibited proliferative and colony-forming capabilities. AM cells differentiated into mature adipocytes, whereas AF cells showed morphological changes but not mature differentiation. These findings confirm the feasibility of collecting and processing AF and AM samples at the time of delivery for stem cells. Although there is variability among AF and AM samples in terms of proliferative and colony-forming capabilities, as well as AM cells differentiation potential. Further research and optimization of methods is needed to better characterize the cell types, including MSCs, present in AF and AM. / 1 / Ann Marie Barfield

The mechanisms which control the accumulation and circulation of the amniotic fluid are poorly understood. Earlier studies had made the tentative suggestion that vasopressin, which is important in the control of water balance in adults, might play a role in the regulation of amniotic fluid turnover. In the work reported here, attempts were made to detect and measure the vasopressin levels in the amniotic fluid of guinea pigs, sheep and humans. Unconcentrated amniotic fluid samples from the guinea pig, sheep and human were assayed using the water-loaded, alcohol-anesthetized rat antidiuretic preparation. If vasopressin is present in the fluid of these animals, it would be at a concentration below the detectable limits of the assay, which were found to be 10 to 15 microunits/ml. Only 4 out of 36 experiments on guinea pig amniotic fluid gave even a suggestion of a response, with 3 of these being too small to be quantified accurately. No sheep or human samples produced an antidiuresis. An attempt was made to dehydrate the maternal guinea pigs, as it. was thought that this would create an osmotic stress in the fetus which might result in increased output of vasopressin. In 29 experiments with fluid of fetuses whose mothers had been without water for 24 or 48 hours prior to collection, 27 gave no response. The responses to the two injections that did indicate activity were too low for accurate quantification. However, these experiments were not considered conclusive, and more extended investigation is needed. Seven human amniotic fluid samples were concentrated on CM-25 Sephadex columns, along with three standard vasopressin loads. The standards showed recoveries of 80% to 90% under optimal conditions. An antidiuretic substance was found in fractions from all of these samples, and it eluted at a similar pH and molarity to the standard vasopressin. This material measured approximately 200 micro-units/ml, and was relatively consistent in all of the samples tested. The amniotic fluid antidiuretic substance (AFAS) appeared to respond with an antidiuresis similar to that of vasopressin on the assay preparation. However, further work suggested that the AFAS could not be identified as vasopressin. Although the pattern of antidiuretic responses were similar, there were marked changes in the sensitivity of the bioassay preparation to the unknown material during periods of little change in the responses to vasopressin itself. A similar contrast was found between different experiments. Sodium thioglycollate incubation also failed to deactivate the active AFAS fractions, whilst control volumes of vasopressin lost apparently all activity following this procedure. The stability characteristics were also dissimilar to those of vasopressin, as the fractions remained active for up to five weeks when stored at neutral pH at 4° C. Optimum pH conditions for the storage of vasopressin are between 3.0 and 5.0. An attempt was made to identify the AFAS as angiotensin II. However, the response pattern on the records of the antidiuretic assay were dissimilar. Further comparisons with combinations of vasopressin and angiotensin II in varying concentrations also failed to mimic the responses to the AFAS. It was concluded that the antidiuretic activity found in the human amniotic fluid was not attributable to vasopressin, angiotensin II, or to a combination of these two hormones, at the levels tested. The nature of the AFAS is at present unknown. Further studies are needed to identify the unknown antidiuretic agent extracted from human amniotic fluid, and the possible mechanisms of its release, and its potential physiological functions, are still unknown. / Science, Faculty of / Zoology, Department of / Graduate

No description available.

The amniotic membrane and the amniotic fluid are one of life's most complex and delicate tissues and fluids, respectively. What was known about this tissue and fluid prior to the 20th century was extremely limited scientifically, but was significantly defined by beliefs entrenched in mysticism, folklore, and superstitions. A comprehensive literature review of the amniotic membrane tissue and amniotic fluid reveals the many unique and complex characteristics and biological properties that been heavily investigated since the turn of the 20th century and continues to surge into the 21st century. The historical perspectives, evolution, derivation, histology, structure, and composition of the amniotic membrane; and historical perspectives, volume and regulation, and cellular and non-cellular composition of the amniotic fluid are discussed here and are coalesced for an easy and comprehensible resource. Lastly, future perspectives regarding research and application of the amniotic membrane and amniotic fluid, including stem cells are discussed.

Liu Bao Yi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 83-104). / Abstracts in English and Chinese. / ABSTRACT --- p.i / ACKNOWLEDGEMENT --- p.v / TABLE OF CONTENTS --- p.vi / LIST OF ABBREVIATIONS --- p.xii / LIST OF FIGURES --- p.xivv / LIST OF TABLES --- p.xv / PUBLICATION RELATED TO THIS THESIS --- p.xvii / Chapter PART 1 --- INTRODUCTION AND LITERATURE RESEARCH / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter CHAPTER 2 --- MECONIUM STAINED LIQUOR --- p.3 / Chapter 2.1 --- AMNIOTIC FLUID --- p.3 / Chapter 2.1.1 --- Function of Amniotic Fluid --- p.3 / Chapter 2.1.2 --- Composition Of Amniotic Fluid --- p.3 / Chapter 2.1.3 --- Regulation Of Amniotic Fluid --- p.4 / Chapter 2.1.4 --- Abnormality Of Amniotic Fluid Volume --- p.4 / Chapter 2.2 --- MECONIUM STAINED LIQUOR --- p.6 / Chapter 2.2.1 --- Formation And Composition Of Meconium --- p.6 / Chapter 2.2.2 --- Peristalsis Of Fetal Gastrointestinal Tract --- p.7 / Chapter 2.2.3 --- Meconium Stained Liquor(MSL) --- p.7 / Chapter 2.2.3.1 --- Maturation Theory --- p.7 / Chapter 2.2.3.2 --- Cord Compression Theory --- p.9 / Chapter 2.2.3.3 --- Fetal Hypoxia Theory --- p.10 / Chapter 2.2.4 --- Fetal Effect Of Meconium In Amniotic Cavity --- p.11 / Chapter 2.2.5 --- Meconium Aspiration Syndrome --- p.12 / Chapter 2.2.6 --- Clinical Significance And Limitation Of Studies --- p.13 / Chapter 2.3 --- Purpose Of Study --- p.14 / Chapter CHAPTER 3 --- OXIDATIVE STRESS AND FETAL HYPOXIA --- p.16 / Chapter 3.1 --- OXIDATIVE STRESS --- p.16 / Chapter 3.2 --- FREE RADICALS --- p.16 / Chapter 3.2.1 --- Sources Of Free Radicals --- p.17 / Chapter 3.2.1.1 --- Biological Source Of Free Radicals --- p.17 / Chapter 3.2.1.2 --- Intracellular Source Of Free Radicals --- p.17 / Chapter 3.2.1.3 --- Composition Of Free Radicals And Reactive Oxygen Species --- p.18 / Chapter 3.2.2 --- Cellular Components At Risk From Free Radicals Damage --- p.20 / Chapter 3.2.2.1 --- Proteins --- p.20 / Chapter 3.2.2.2 --- Nucleic Acids And DNA --- p.21 / Chapter 3.2.2.3 --- Membrane Lipids --- p.21 / Chapter 3.2.3 --- Lipid Peroxidation --- p.21 / Chapter 3.2.3.1 --- Chemical Substances Of Membranes --- p.21 / Chapter 3.2.3.2 --- The Reactions Of Lipid Peroxidation --- p.22 / Chapter 3.2.3.3 --- Lipid Peroxidation In Pregnancy --- p.23 / Chapter 3.2.4 --- Protection Against Lipid Peroxidation --- p.24 / Chapter 3.2.5 --- Isoprostanes --- p.26 / Chapter 3.2.5.1 --- Definition --- p.26 / Chapter 3.2.5.2 --- Formation Of Isoprostanes --- p.26 / Chapter 3.2.5.3 --- Metabolism Of Isoprostanes --- p.27 / Chapter 3.2.5.4 --- Biological Characteristics Of Isoprostanes --- p.29 / Chapter 3.2.5.5 --- Isoprostanes As Mediators Of Oxidantive Stress --- p.29 / Chapter 3.3 --- FETAL HYPOXIA --- p.30 / Chapter 3.3.1 --- Fetal Metabolism And Energy Supply --- p.30 / Chapter 3.3.2 --- Free Radical Generation And Fetal Hypoxia-Reoxygenation --- p.33 / Chapter 3.3.3 --- Fetal Hypoxia And Fetal Brain Injury --- p.34 / Chapter 3.3.4 --- Measurement Of Fetal Hypoxia --- p.35 / Chapter 3.3.4.1 --- Acid-Base Balance --- p.35 / Chapter 3.3.4.2 --- Fetal Heart Rate Monitoring --- p.36 / Chapter 3.3.4.3 --- Apgar scores --- p.37 / Chapter 3.3.4.4 --- Pulse Oximetry --- p.37 / Chapter 3.3.4.5 --- Lipid Peroxides --- p.38 / Chapter CHAPTER 4 --- AMNIOINFUSION --- p.40 / Chapter 4.1 --- AMNIOINFUSION --- p.40 / Chapter 4.2 --- AMNIOINFUSION FOR OLIGOHYDRAMNIOS --- p.40 / Chapter 4.3 --- AMNIOINFUSION FOR MECONIUM STAINED LIQUOR --- p.41 / Chapter 4.4 --- PURPOSE OF THE STUDY --- p.42 / Chapter PART 2 --- CLINICAL PROTOCOLS AND MEASUREMENT OF ISOPROSTANES / Chapter CHAPTER 5 --- CLINICAL PROTOCOLS --- p.43 / Chapter 5.1 --- ETHICS --- p.43 / Chapter 5.2 --- CLINICAL PROTOCOLS --- p.43 / Chapter 5.2.1 --- Artificial Rupture Of Membranes (Amniotomy) --- p.43 / Chapter 5.2.2 --- Classification of Meconium Stained Liquor --- p.44 / Chapter 5.2.3 --- Electronic Fetal Heart Rate Monitoring --- p.44 / Chapter 5.2.4 --- Monitoring The Progress of Labour --- p.44 / Chapter 5.2.5 --- Umbilical Cord Blood Gas Analysis --- p.45 / Chapter 5.2.6 --- Apgar Score --- p.45 / Chapter 5.2.7 --- Meconium Aspiration --- p.46 / Chapter 5.2.8 --- Clinical Outcome --- p.46 / Chapter CHAPTER 6 --- MEASUREMENT OF ISOPROSTANES --- p.50 / Chapter 6.1 --- BLOOD PREPARATION --- p.50 / Chapter 6.2 --- REAGENTS --- p.50 / Chapter 6.3 --- GAS CHROMATOGRAPHY AND MASS SPECTROMETRY (GC-MS) --- p.51 / Chapter 6.4 --- PROCEDURES --- p.51 / Chapter 6.5 --- DATA RELIABILITY --- p.53 / Chapter PART 3 --- RESULTS AND DISCUSSION / Chapter CHAPTER 7 --- MECONIUM STAINED LIQUOR (MSL) DURING LABOUR AND NEONATAL CORD BLOOD 8-IS〇-PGF2α CONCENTRATION --- p.54 / Chapter 7.1 --- OBJECTIVE --- p.54 / Chapter 7.2 --- MATERIALS AND METHOD --- p.55 / Chapter 7.3 --- STATISTICAL ANALYSIS --- p.56 / Chapter 7.4 --- RESULTS --- p.57 / Chapter 7.5 --- DISCUSSION --- p.65 / Chapter 7.6 --- CONCLUSION --- p.67 / Chapter CHAPTER 8 --- EVALUATION OF PROPHYLACTIC AMNIOINFUSION FOR INTRAPARTUM MECONIUM STAINED LIQUOR --- p.69 / Chapter 8.1 --- OBJECTIVE --- p.69 / Chapter 8.2 --- MATERIALS AND METHOD --- p.69 / Chapter 8.2.1 --- Study Group: 226}0ب MSL+AI' --- p.69 / Chapter 8.2.2 --- The Procedure Of Amnioinfusion --- p.70 / Chapter 8.2.3 --- Other Study Group --- p.71 / Chapter 8.3 --- STATISTIC ANALYSIS --- p.71 / Chapter 8.4 --- RESULTS --- p.72 / Chapter 8.4.1 --- Comparison Between The 'MSL+AI' And 'MSL-AI' Groups --- p.72 / Chapter 8.4.2 --- Comparison Between 226}0بMSL+AI'And 'Clear Liquor' Groups --- p.74 / Chapter 8.5 --- DISCUSSION --- p.77 / Chapter 8.6 --- CONCLUSION --- p.79 / Chapter CHAPTER 9 --- COMMENTS AND FUTURE RESEARCH --- p.80 / BIBLIOGRAPHY --- p.83

This study was designed to examine the hypothesis that the amount of amniotic fluid present during gestation is critical to normal lung growth and maturation. On day 16 of gestation the amniotic sacs of the right or left uterine horns of timed pregnant Sprague-Dawley rats were punctured with a 20 gauge needle. The fetuses of the opposite horn served as controls. On day 21 of gestation (one day prior to natural delivery) the fetuses were delivered by Cesarean section. An unbalanced, mixed model analysis of variance was performed on the data collected from each fetus. Probability values of less than 0.05 between control and experimental animals were considered significant. Amniotic sac puncture resulted in a significant loss of amniotic fluid as indicated by reduced amniotic fluid volume on day 21. Experimental body weight was significantly reduced indicating fetal growth retardation. Lung growth was also retarded as indicated by significantly reduced lung weight to body weight ratios and lung volume to body weight ratios following amniotic sac puncture. There was a reduction in the amount of fluid present within the experimental lungs. There appeared to be no significant effect on the structural units of the lung as indicated by no significant difference between control and experimental fetal lungs in terms of cell number, cell size, total protein to body weight ratio, maturation of type II cells, volume fraction of saccular air, saccular wall, conducting air and nonparenchyma, airspace size, saccular surface area to body weight ratio and surface to volume ratio. Thus, loss of amniotic fluid significantly affected lung growth, more than it affected overall body growth, without having an effect on lung maturation. / Medicine, Faculty of / Pathology and Laboratory Medicine, Department of / Graduate

Our objectives were to measure concentrations of glucose, insulin, insulin-like-growth-factor-binding-protein-1 (IGF BP1) and beta-hydroxybutyrate (BOHB) in amniotic fluid (AF), and establish if these concentrations were associated with emergence of maternal gestational diabetes mellitus (GDM). AF samples (n=408) were collected following routine amniocentesis (12-22 weeks gestation). Glucose and insulin concentrations were elevated in our GDM mother-infant pairs, where GDM was associated with a 176g increase in birth weight. Logistic regression showed that AF glucose but not insulin was associated with developing GDM. Non-linear Bayesian probability plots showed that when 2nd trimester glucose was plotted against insulin increases in both were predictive of the subsequent emergence of GDM. In conclusion, our findings show that: (1) AF glucose but not insulin predicts subsequent emergence of GDM and (2) these observed elevations provide evidence that the fetus of GDM mothers is being exposed early in-utero to metabolic perturbations (i.e. elevated glucose) that may have important long-term metabolic consequences for their future development.

Made available in DSpace on 2014-06-11T19:23:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-06-05Bitstream added on 2014-06-13T20:30:41Z : No. of bitstreams: 1 donatti_c_me_botfmvz.pdf: 2093911 bytes, checksum: d264970227347791e8738946cc4ac63c (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A córnea é a estrutura anterior transparente do olho e apresenta-se muito susceptível a traumas e agressões. O objetivo do presente estudo foi avaliar e comparar, clínica e histomorfometricamente, o processo de reparação corneana de úlceras induzidas em coelhos, frente à utilização do plasma rico em plaquetas (PRP) sob a forma de colírio ou tampão. Foram utilizadas 60 fêmeas da espécie leporina, constituindo-se 4 grupos experimentais de 15 animais cada, designados grupo plaquetas (GP), grupo tampão (GT), grupo controle (GC) e grupo controle amniótica (GA). Em todos os animais foi realizada a úlcera experimental, sendo este o único procedimento no GC. No GP, os coelhos foram medicados com colírio autólogo de plasma rico em plaquetas. No GC, foi instilado colírio lubrificante. No GT, foi aplicado tampão sólido rico em plaquetas, revestido por membrana amniótica, para a retenção do mesmo. No GA, foi aplicada apenas a membrana amniótica. Os grupos experimentais foram subdivididos em três subgrupos (M4, M7, M30), de acordo com os períodos finais de avaliação. Os animais foram avaliados por meio de exame clínico e histomorfométrico. Não houve diferenças entre os tratamentos utilizados quanto aos sinais relacionados à sensibilidade ocular, quemose e secreção ocular. Os grupos tratados com PRP, na forma de tampão ou colírio, apresentaram menor opacidade do que os animais tratados apenas com membrana amniótica no momento final de avaliação. Quanto à presença da úlcera corneana, os grupos tratados com PRP (GP e GT) apresentaram menor intensidade de ulceração corneana com relação aos demais grupos (GC e GA). Na avaliação histológica verificou-se maior epitelização corneana na fase inicial da lesão no tratamento à base de colírio de PRP. O uso da membrana amniótica promoveu espessamento do epitélio e estroma corneano, com sinergismo da mesma quando associada ao PRP / The cornea is the transparent anterior part of the eye and is very susceptible to trauma and sore. The aim of this work was to evaluate and compare both, clinically and histomorphometrically the process of repair of induced corneal ulcer in rabbits using platelet-rich plasma in the form of eyedrop or clot. Sixty female leporids were divided into four groups of 15 animals, and named as platelet group (PG), clot group (CLG), control group (CG), and amniotic control group (AG). Ocular ulcer was experimentally induced in all the animals. Ulcer induction was the single procedure performed in CG. In PG, autologous platelet-rich plasma as a eyedrop was used five times a day for seven days. In CLG, a platelet-rich clot was covered by amniotic membrane to hold it in place was used. In AG, only the amniotic membrane was used. Experimental groups were then subdivided into three groups (M4, M7, M30), corresponding to the end of the evaluation period. The animals were evaluated through clinical and histomorphometric tests. There were no differences between treatments related to ocular sensitivity (blepharospasm and photophobia), chemosis and ocular secretion. The groups treated with PRP either as eyedrop or a clot, showed less opacity than the animals treated only with amniotic membrane at the moment of the final evaluation. The presence of corneal ulcers in the groups treated with PRP (PG and CLG) showed lower intensity than the other groups (CG and AG). In histological evaluation, corneal epithelization at the initial phase of the lesion was confirmed when using PRP. The use of amniotic membrane promoted corneal epithelial and stromal thickness, as well as synergism when associated to PRP

No description available.

Amniotic fluid (AF) is a protective pool and a resource of amino acids for the growing fetus. In study 1, we investigated if any of these AF amino acids at mid gestation were associated with fetal development in humans. Nineteen amino acids differed across birth weight percentiles. Arginine, 3-methyl histidine and tryptophan were positive predictors of birth weight, while ornithine was a negative predictor. In study 2, we used a diet induced model of IUGR to see if specific AF amino acids were predictive of fetal weight near term. Methionine and phenylalanine were modified by diet, and 12 amino acids were independently modified by gestational age, respectively. Cysteine, lysine, methionine and tyrosine were predictors of fetal weight. Thus, the AF amino acid pool is associated in animals and humans with fetal growth.