Medicine and Health Sciences Commons^™

Prenatal Genetic Diagnosis Using Transcervically Derived And Immunomagnetically Isolated Trophoblast Cells, Swati Bajpayee

Honors College Theses

Trophoblast cells migrate from the placenta into the endocervical canal early in the first trimester and can be collected non-invasively by transcervical sampling (TCS), potentially providing fetal DNA for prenatal genetic diagnosis. Experiments were conducted to separate fetal cells from maternal cells within the TCS specimens and use the fetal cells to perform genetic analysis. Trophoblast cells were efficiently isolated from TCS specimens during weeks 5-18 of gestation using HLA-G antibody coupled to magnetic nanoparticles. Immunofluorescence microscopy revealed that 99% of isolated cells expressed the chorionic gonadotropin β subunit (β-CG), while the non-bound cell fraction expressed none. Immunomagnetically isolated cells …

Preterm Birth And The Perception Of Risk Among African Americans, Gwendolyn Simpson Norman

Wayne State University Dissertations

Background: African American women deliver preterm at a rate that is two to three times that of their white counterparts, and after decades of research, this disparity in birth outcomes still remains unexplained. While factors including income, education, neighborhood conditions, infection and stress have all been associated with prematurity, no combination of these factors has explained why the disparity persists. Recently, however, racism-specific stress has emerged as a possible factor contributing to this disparity. This study was designed to learn how preterm birth was explained by African Americans directly impacted by prematurity. Methods: Interviews were conducted with African American women …

Does Cerclage Improve Neonatal Outcomes In A Molar Pregnancy And A Coexistent Fetus? A Case Report, Eduardo Aguin, Victor Aguin, Ligia Cisneros, Tina Aguin, Cosmas Van De Ven, Ray Bahado-Singh

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

Complete hydatiform mole and coexistent viable fetus is very rare. The use of a cervical cerclage for cervical indications in the presence of this condition has never been reported. Although the diagnosis was made postnatal, the objective is to present a case with good neonatal outcome.

Case presentation

A patient presented with vaginal spotting around 23 weeks. She has a history of four preterm deliveries. Her cervix was dilated and a cerclage was placed. She presented again with PPROM around 25 weeks. She went into spontaneous preterm labor and delivered a viable fetus that is a healthy girl …

Fetal Erap2 Variation Is Associated With Preeclampsia In African Americans In A Case-Control Study, Lori D. Hill, Dashaunda D. Hilliard, Timothy P. York, Sindhu Srinivas, Juan P. Kusanovic, Ricardo Gomez, Michal A. Elovitz, Roberto Romero, Jerome F. Strauss Iii

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

Preeclampsia affects 3-8% of pregnancies and is a major cause of maternal and perinatal morbidity and mortality worldwide. This complex disorder is characterized by alterations in the immune and vascular systems and involves multiple organs. There is strong evidence for a genetic contribution to preeclampsia. Two different single nucleotide polymorphisms (SNPs) in the endoplasmic reticulum aminopeptidase 2 (ERAP2) gene were recently reported to be associated with increased risk for preeclampsia in two different populations. ERAP2 is expressed in placental tissue and it is involved in immune responses, inflammation, and blood pressure regulation; making it is an attractive preeclampsia …

The Potential Role Of Innate Immunity In The Pathogenesis Of Post-Operative Adhesions, Jennell White

Wayne State University Dissertations

Post-operative adhesion development occurs in the vast majority of patients following abdominal surgery and is a natural occurrence of peritoneal-wound healing. These fibrous bands may form within the first 5-7 days post-surgery and have the ability to cause a distortion in the normal anatomical positioning of abdominal organs. Consequently, adhesions are major contributors to small bowel obstruction, infertility, and severe pelvic and abdominal pain. Physiological processes responsible for adhesion formation remain obscure though it is believed to involve cell migration, proliferation, and differentiation of several cell types including mesothelial cells, fibroblasts, and inflammatory and immune cells. Substances released from these …

Human Trophoblast Survival And Invasion In The Developing Placenta: Autocrine Regulation By Hbegf, Philip Jessmon

Wayne State University Dissertations

HBEGF is a multifunctional protein in early pregnancy that induces cytotrophoblast (CTB) cell differentiation to an invasive phenotype, protects against apoptosis, and is involved in an autocrine signaling mechanism that leads to its own protein synthesis. CTBs exist in a low O₂ environment during the first 10 weeks of implantation, during which they invade the decidualized uterine stroma. Inhibitors of intracellular signaling pathways demonstrated that at 20% O₂ HBEGF induces an increase in cell migration through the ERK, MAPK14, JNK, or PIK3 pathways downstream of signaling through its ERBB receptors. Also downstream of these four pathways, HBEGF induces …

Sulforaphane Induces Cell Cycle Arrest By Protecting Rb-E2f-1 Complex In Epithelial Ovarian Cancer Cells, Christopher S. Bryant, Sanjeev Kumar, Sreedhar Chamala, Jay Shah, Jagannath Pal, Mahdi Haider, Shelly Seward, Aamer M. Qazi, Robert Morris, Assaad Semaan, Masood A. Shammas, Christopher Steffes, Ravindra B. Potti, Madhu Prasad, Donald W. Weaver, Ramesh B. Batchu

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

Sulforaphane (SFN), an isothiocyanate phytochemical present predominantly in cruciferous vegetables such as brussels sprout and broccoli, is considered a promising chemo-preventive agent against cancer. In-vitro exposure to SFN appears to result in the induction of apoptosis and cell-cycle arrest in a variety of tumor types. However, the molecular mechanisms leading to the inhibition of cell cycle progression by SFN are poorly understood in epithelial ovarian cancer cells (EOC). The aim of this study is to understand the signaling mechanisms through which SFN influences the cell growth and proliferation in EOC.

Results

SFN at concentrations of 5 - 20 …

Nuclear Factor-Kappa B Localization And Function Within Intrauterine Tissues From Term And Preterm Labor And Cultured Fetal Membranes, Sonali Vora, Asad Abbas, Chong J. Kim, Taryn Ls Summerfield, Juan P. Kusanovic, Jay D. Iams, Roberto Romero, Douglas A. Kniss, William E. Ackerman Iv

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

The objective of this study was to quantify the nuclear localization and DNA binding activity of p65, the major transactivating nuclear factor-kappa B (NF-kappaB) subunit, in full-thickness fetal membranes (FM) and myometrium in the absence or presence of term or preterm labor.

Methods

Paired full-thickness FM and myometrial samples were collected from women in the following cohorts: preterm no labor (PNL, N = 22), spontaneous preterm labor (PTL, N = 21), term no labor (TNL, N = 23), and spontaneous term labor (STL, N = 21). NF-kappaB p65 localization was assessed by immunohistochemistry, and DNA binding activity was …

Ritonavir Blocks Akt Signaling, Activates Apoptosis And Inhibits Migration And Invasion In Ovarian Cancer Cells, Sanjeev Kumar, Christopher S. Bryant, Sreedhar Chamala, Aamer Qazi, Shelly Seward, Jagannath Pal, Christopher P. Steffes, Donald W. Weaver, Robert Morris, John M. Malone, Masood A. Shammas, Madhu Prasad, Ramesh B. Batchu

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

Ovarian cancer is the leading cause of mortality from gynecological malignancies, often undetectable in early stages. The difficulty of detecting the disease in its early stages and the propensity of ovarian cancer cells to develop resistance to known chemotherapeutic treatments dramatically decreases the 5-year survival rate. Chemotherapy with paclitaxel after surgery increases median survival only by 2 to 3 years in stage IV disease highlights the need for more effective drugs. The human immunodeficiency virus (HIV) infection is characterized by increased risk of several solid tumors due to its inherent nature of weakening of immune system. Recent observations …

Sensitization Of Ovarian Cancer Cells To Cisplatin By Genistein: The Role Of Nf-Kappab, Leigh A. Solomon, Shadan Ali, Sanjeev Banerjee, Adnan R. Munkarah, Robert T. Morris, Fazlul H. Sarkar

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

Platinum-resistance (PR) continues to be a major problem in the management of epithelial ovarian cancer (EOC). Response to various chemotherapeutic agents is poor in patients deemed PR. Genistein, a soy isoflavone has been shown to enhance the effect of chemotherapy in prostate and pancreatic cancer cells in vitro and in vivo by reversing chemo-resistance phenotype. The goal of this study was to investigate the effects of combination therapy with genistein and cisplatin as well as other cytotoxic conventional chemotherapeutic agents in platinum-sensitive (PS) and resistant EOC cells.

Methods

The PS human ovarian cancer cell line A2780 and its …

Using Hyperosmolar Stress To Measure Biologic And Stress-Activated Protein Kinase Responses In Preimplantation Embryos, Yufen Xie, W Zhong, Y Wang, A Trostinskaia, F Wang, Elizabeth E. Puscheck, D. A. Rappolee

Department of Obstetrics and Gynecology Research Publications

We used hyperosmolar stress to test blastocysts for their biologic and enzymatic responses to culture stress. Embryos mount dose- and time-dependent responses to hyperosmolar stress. Biological responses included slowed cavitation and cell accumulation and increased apoptosis at increasing doses. These responses were preceded by stress-activated protein kinase (SAPK) phosphorylation and nuclear translocation consistent with its causal role. For cavitation and new cell cycle initiation, 200 mM sorbitol caused stasis. Above 200 mM, sorbitol was ultimately lethal and below 200 mM, its embryos had milder effects. Phosphorylated SAPK was induced rapidly in embryos at 0.5 h in a dose-dependent manner from …

Effects Of Sapk/Jnk Inhibitors On Preimplantation Mouse Embryo Development Are Influenced Greatly By The Amount Of Stress Induced By The Media, Yufen Xie, E. E. Puscheck, D. A. Rappolee

Department of Obstetrics and Gynecology Research Publications

Stress-activated protein kinase/c-Jun kinase (SAPK/JNK) is thought to be necessary for preimplantation embryonic development (Maekawa et al., 2005). However, media increases SAPK/JNK phosphorylation and these levels negatively correlate with embryonic development (Wang et al., 2005). Culture-induced stress could confuse analysis of the role of SAPK in development. In this study, we tested how SAPK/JNK inhibitors influence embryonic development in optimal and non-optimal media and define the contribution of cell survival and proliferation to the embryonic response to these media. SAPK/JNK inhibitors retard embryonic development in suboptimal Ham’s F10, but improve development in optimal potassium (K+) simplex optimized media (KSOM) +AA. …

The Role Of Aromatase Inhibitors In Ameliorating Deleterious Effects Of Ovarian Stimulation On Outcome Of Infertility Treatment, Mohamed Fm Mitwally, Robert F. Casper, Michael P. Diamond

Wayne State University Associated BioMed Central Scholarship

Abstract

Clinical utilization of ovulation stimulation to facilitate the ability of a couple to conceive has not only provided a valuable therapeutic approach, but has also yielded extensive information on the physiology of ovarian follicular recruitment, endometrial receptivity and early embryo competency. One of the consequences of the use of fertility enhancing agents for ovarian stimulation has been the creation of a hyperestrogenic state, which may influence each of these parameters. Use of aromatase inhibitors reduces hyperestrogenism inevitably attained during ovarian stimulation. In addition, the adjunct use of aromatase inhibitors during ovarian stimulation reduces amount of gonadotropins required for optimum …

Serine-Threonine Kinases And Transcription Factors Active In Signal Transduction Are Detected At High Levels Of Phosphorylation During Mitosis In Preimplantation Embryos And Trophoblast Stem Cells, Jian Liu, Elizabeth E. Puscheck, Fangfei Wang, Anna Trostinskaia, Dusan Barisic, Gordon Maniere, Dana Wygle, W Zhong, Edmond Hhm Rings, Daniel A. Rappolee

Department of Obstetrics and Gynecology Research Publications

Serine-threonine kinases and transcription factors play important roles in the G1-S phase progression of the cell cycle. Assays that use quantitative fluorescence by immunocytochemical means, or that measure band strength during Western blot analysis, may have confused interpretations if the intention is to measure G1-S phase commitment of a small subpopulation of phosphorylated proteins, when a larger conversion of the same population of proteins can occur during late G2 and M phases. In mouse trophoblast stem cells (TSC), a human placental cell line (HTR), and/or mouse preimplantation embryos, 8/19 ser- ine-threonine and tyrosine kinases, 3/8 transcription factors, and 8/14 phospho …

Human Chorionic Gonadotropin Administration Is Associated With High Pregnancy Rates During Ovarian Stimulation And Timed Intercourse Or Intrauterine Insemination, Mohamed F. Mitwally, Sonya Abdel-Razeq, Robert F. Casper

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

There are different factors that influence treatment outcome after ovarian stimulation and timed-intercourse or intrauterine insemination (IUI). After patient age, it has been suggested that timing of insemination in relation to ovulation is probably the most important variable affecting the success of treatment. The objective of this study is to study the value of human chorionic gonadotropin (hCG) administration and occurrence of luteinizing hormone (LH) surge in timing insemination on the treatment outcome after follicular monitoring with timed-intercourse or intrauterine insemination, with or without ovarian stimulation.

Methods

Retrospective analysis of 2000 consecutive completed treatment cycles (637 timed-intercourse and …

Prevalence And Risk Factors For Vaginal Candida Colonization In Women With Type 1 And Type 2 Diabetes, Ella M. De Leon, Scott J. Jacober, Jack D. Sobel, Betsy Foxman

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

Diabetes mellitus increases the rate of vaginal colonization and infection with Candida species

Methods

We surveyed women with diabetes receiving care at either an urban or suburban diabetes clinic to examine the relationship between vaginal Candida colonization, diabetes type and duration, and HbA_1c level. 101 participants completed the self-administered questionnaire and self-collected a vaginal swab for Candida culture. Candida colonization was similar by age and race.

Results

Type 1 diabetics were three times as likely as type 2 diabetics to be colonized with any Candida species (OR = 3.4; 95% CI: 1.03, 11.41; p = 0.04); even …

Basement Membrane And Repair Of Injury To Peripheral Nerve: Defining A Potential Role For Macrophages, Matrix Metalloproteinases, And Tissue Inhibitor Of Metalloproteinases-1, Monique La Fleur, Johnnie L. Underwood, Daniel A. Rappolee, Zena Werb

Department of Obstetrics and Gynecology Research Publications

Injury to a peripheral nerve is followed by a remodeling process consisting of axonal degenera- tion and regeneration. It is not known how Schwann cell–derived basement membrane is pre- served after injury or what role matrix metalloproteinases (MMPs) and their inhibitors play in axonal degeneration and regeneration. We showed that the MMPs gelatinase B (MMP-9), stromelysin-1 (MMP-3), and the tissue inhibitor of MMPs (TIMP)-1 were induced in crush and distal segments of mouse sciatic nerve after injury. TIMP-1 inhibitor activity was present in excess of proteinase activity in extracts of injured nerve. TIMP-1 protected basement mem- brane type IV collagen …

Hepatocyte Growth Factor And Its Receptor Are Expressed In Cardiac Myocytes During Early Cardiogenesis, Daniel A. Rappolee, Anand Iyer, Yogesh Patel

Department of Obstetrics and Gynecology Research Publications

In the mouse, the heart primordium arises when mesodermis set aside during gastrulation, is induced by pharyngeal endoderm, migrates ventrally to the midline of the embryo, forms a tube, and begins beating. Little is known of the molecular mechanisms that mediate the determination, mitosis, differentiation, and migration that lead to the beating heart. Transcripts for hepatocyte growth factor/scatter factor (HGF) and its receptor are coexpressed transiently and dynamically in the premyocardium but not in other heart progenitor cells. Transcripts for HGF ligand and receptor are first detected before cardiac function and looping and persist through the first looping stage, when …

Expression And Function Of Fgf-4 In Peri-Implantation Development In Mouse Embryos, Daniel A. Rappolee, Claudio Basilico, Yogesh Patel, Zena Werb

Department of Obstetrics and Gynecology Research Publications

One of the earliest events in mammalian embryogenesis is the formation of the inner cell mass (ICM) and the subse- quent delamination of primitive endoderm. We have found that mRNA for fibroblast growth factor (FGF)-4, but not FGF-3, is expressed in preimplantation mouse blastocysts and that the FGF-4 polypeptide is present in ICM cells. ICM-like embryonal carcinoma cells and embryonic stem cells also express FGF-4. Conversely, differentiated embryonal carcinoma cells in the endoderm lineage express FGF-3, but not FGF-4 mRNA. Although mouse embryos expressed FGF-4 mRNA from the 1-cell stage, embryos cultured from the 2-cell through the blastocyst stage in …

Insulin-Like Growth Factor Ii Acts Through An Endogenous Growth Pathway Regulated By Imprinting In Early Mouse Embryos, Daniel A. Rappolee, Karin S. Sturm, Ole Behrendtsen, Gilbert A. Schultz, Roger A. Pedersen, Zena Werb

Department of Obstetrics and Gynecology Research Publications

We present evidence that insulin-like growth factor II (IGF-II) mediates growth in early mouse embryos and forms a pathway in which imprinted genes influence development during preimplantation stages, mRNA and protein for IGF-II were expressed in preimplantation mouse embryos, but the related factors IGF-I and insulin were not. IGF-I and insulin receptors and the IGF-II/mannose-6-phosphate receptor were expressed. Exogenous IGF-II or IGF-I increased the cell number in cultured blastocysts, but a mutant form of IGF-II that strongly binds only the IGF-II receptor did not. Reduction of IGF-II expression by antisense IGF-II oligonucleotides decreased the rate of progression to the blastocyst …

Platelet A-Granule Fibrinogen, Albumin, And Immunoglobulin G Are Not Synthesized By Rat And Mouse Megakaryocytes, Prem Handagama, Daniel A. Rappolee, Zena Werb, Jack Levin, Dorothy F. Bainton

Department of Obstetrics and Gynecology Research Publications

It has been assumed that endogenous synthesis by the platelet precursor cell, the bone marrow megakaryocyte, is the major source of platelet a-granule protein. To test this hypothesis, we used mRNA phenotyping to detect in megakaryocytes the presence of mRNA transcripts specific for various proteins. Our results indicate that megakaryocytes synthesize platelet factor 4, a protein relatively specific for platelets, but do not express mRNA transcripts for the fibrinogen, albumin, or IgG found in a-granules. We have previously shown that mega- karyocytes endocytose circulating proteins, including fibrino- gen, albumin, and IgG, and incorporate them into a-granules. Thus, platelets appear to …

Two Alkaline Phosphatase Genes Are Expressed During Early Development In The Mouse Embryo, Ann C. Hahnel, Daniel A. Rappolee, Jose Luis Millan, Thomas Manes, Carol A. Ziomek, Nicoleta G. Theodosiou, Zena Werb, Roger A. Pedersen, Gilbert A. Schultz

Department of Obstetrics and Gynecology Research Publications

Alkaline phosphatase (AP) activity is stage specific in mouse embryos and may be associated with compaction and separation of trophectoderm from inner cell mass in preimplantation development. We previously sequenced a cDNA and two mouse AP genes that could contribute to the AP activity in embryos. Oligonucleotide primers were constructed from the three sequences and used in the reverse transcription-polymerase chain reaction technique to establish that two of the three AP isozymes are transcribed during preimplantation development. The predominant transcript (E-AP) is from a gene highly homologous to the human tissue-specific APs, but different from the mouse intestinal AP. Tissue …

Genes For Extracellular Matrix-Degrading Metalloproteinases And Their Inhibitor, Timp, Are Expressed During Early Mammalian Development, C. A. Brenner, R. R. Adler, D. A. Rappolee, R. A. Pedersen, Z Werb

Department of Obstetrics and Gynecology Research Publications

Extracellular matrix (ECM) remodeling accompanies cell migration, cell-cell interactions, embryo expansion, uterine implantation, and tissue invasion during mammalian embryogenesis. We have found that mouse embryos secrete functional ECM-degrading metalloproteinases, including collagenase and stromelysin, that are inhibitable by the tissue inhibitor of metalloproteinases (TIMP) and that are regulated during peri-implantation development and endoderm differentiation. mRNA transcripts for collagenase, stromelysin, and TIMP were detected as maternal transcripts in the unfertilized egg, were present at the zygote and cleavage stages, and increased at the blastocyst stage and with endoderm differentiation. These data suggest that metalloproteinases function in cell-ECM interactions during growth, development, and …

Developmental Expression Of Pdgf, Tgf-A, And Tgf-J Genes In Preimplantation Mouse Embryos, Daniel A. Rappolee, Carol A. Brenner, Richard Schultz, David Mark, Zena Werb

Department of Obstetrics and Gynecology Research Publications

Control of growth and differentiation during mammalian embryogenesis may be regulated by growth factors from embryonic or maternal sources. With the use of single-cell messenger RNA phenotyping, the simultaneous expression of growth factor transcripts in single or small numbers of preimplantation mouse embryos was examined. Transcripts for platelet-derived growth factor A chain (PDGF-A), trans- forming growth factor (TGF)-a, and TGF- 1, but not for four other growth factors, were found in whole blastocysts. TGF-cx, TGF-(31, and PDGF antigens were detected in blastocysts by immunocytochemistry. Both PDGF-A and TGF-a were detected as maternal transcripts in the unferilized ovulated oocyte, and again …