Roohi Jeelani

Trophoblast retrieval and isolation from the cervix (TRIC) for noninvasive prenatal screening at 5 to 20 weeks of gestation

OBJECTIVE To use trophoblast cells accumulating in the endocervical canal at the beginning of pregnancy for noninvasive prenatal testing. DESIGN Prospective, double-blinded test for fetal gender. SETTING Academic medical center. PATIENT(S) Fifty-six women with singleton pregnancies at gestational age 5-20 weeks. INTERVENTION(S) Isolation of fetal cells from resident maternal cells in endocervical specimens using anti-human leukocyte antigen G coupled to magnetic nanoparticles; cell phenotyping immunofluorescently with a panel of trophoblast subtype-specific proteins; DNA integrity assessment with terminal dUTP nick-end labeling (TUNEL); and polymerase chain reaction (PCR) and fluorescent in situ hybridization (FISH) to detect sex chromosomes in individual cells. MAIN OUTCOME MEASURE(S) Trophoblast phenotype, TUNEL index, and percentage male cells. RESULT(S) The women were given a routine Papanicolaou test; fetal genders were verified from medical records. Recovery after immunomagnetic isolation averaged 746±59 cells across gestational age, with 99% expressing chorionic gonadotropin, whereas the depleted cell fraction expressed none. The isolated cells had an extravillous trophoblast phenotype and intact nuclear DNA (>95%). Fetal gender was determined in 20 specimens without error by PCR. The FISH analysis of isolated cells from male specimens validated their fetal origin. CONCLUSION(S) Noninvasive prenatal testing is feasible beginning at a gestational age of 5 weeks.

Preimplantation genetic diagnosis for inherited neurological disorders

Preimplantation genetic diagnosis (PGD) is an option for couples at risk of having offspring with an inherited debilitating or fatal neurological disorder who wish to conceive a healthy child. PGD has been carried out for conditions with various modes of inheritance, including spinal muscular atrophy, Huntington disease, fragile X syndrome, and chromosomal or mitochondrial disorders, and for susceptibility genes for cancers with nervous system involvement. Most couples at risk of transmitting a genetic mutation would opt for PGD over prenatal testing and possible termination of a pregnancy. The aim of this Perspectives article is to assist neurologists in counselling and treating patients who wish to explore the option of PGD to enable conception of an unaffected child. PGD can be accomplished for most disorders in which the genetic basis is known, and we argue that it is time for clinicians and neurological societies to consider the evidence and to formulate guidelines for the responsible integration of PGD into modern preventative neurology.

Peroxynitrite deteriorates oocyte quality through disassembly of microtubule organizing centers

Previous theoretical studies have suggested that utilization of 3-D imaging to acquire morphologic parameters of meiotic spindles may be useful in infertility related procedures as an assessment of oocyte quality. However, our results show that treatment of oocytes with increasing concentrations of peroxynitrite (ONOO(-)) caused a dramatic alteration in spindle shape in which morphologic parameters are not measurable or are uninformative in terms of oocyte quality. Metaphase II mouse oocytes (n=520) were treated with increasing concentrations of ONOO(-), after which all oocytes were fixed and subjected to indirect immunofluorescence. Oocyte quality was assessed by alterations in the microtubule-organizing center (MTOC), pericentrin location, microtubule morphology, and chromosomal alignment. In untreated oocytes, pericentrin is primarily assembled utilizing the acentrosomal MTOC, which appears as a condensation at both spindle poles. The spindle has a symmetrical pointed barrel shape, assembled around the chromosomal plate at the spindle equator. Oocytes treated with low concentrations of ONOO(-) (<2.5 μM) showed shortening of the spindle apparatus, while pericentrin scatters from a tight condensation to a dispersed cluster around each spindle pole. At higher ONOO(-) concentrations (>2.5μM) the central attachments between microtubules are strained and bend or unevenly break, and the MTOC proteins are further dispersed or undetectable. Peroxynitrite mediated MTOC damage, which deranges the chromosomal scaffold at the time of assembly and separation, caused the deterioration in oocyte quality. These results provide a link between reactive oxygen species and poor reproductive outcomes and elucidate the underlying etiology, which could be used as a superior biomarker for oocyte quality compared to existing assessment tools.

Maternal race and neonatal outcomes after elective repeat cesarean delivery

Abstract Objective: To determine the effect of race in the risks of prematurity-related complications (PRC) after elective repeat cesarean delivery (ERCD). Methods: The NCHS-CDC Database for the U.S. (2004–2008) was used. ERCD cases were included. Exclusion criteria were multiple gestation, trial of labor, fetal anomalies, history of diabetes and/or hypertension. PRC analyzed were: Apgar score, assisted ventilation, intensive care admission, surfactant use, antibiotic use, seizures. Regression analysis was performed to calculate the odds ratio (OR) of these variables. Deliveries at 36–40 weeks were studied with 40 weeks as reference. Results: Totally, 785 340 ERCDs were performed between 36 and 40 weeks. For the overall population, there was not difference in adverse outcomes between 39 and 40 weeks. The rates of PRC were significantly higher in newborns at 38 compared to 39 weeks, with similar findings in sub-analysis of whites. For African-Americans, the rate of PRC was not significantly different at 38 compared to 39 weeks. Conclusions: We report increased rates of PRC after ERCD before 39 weeks, similar findings from smaller hospital-based studies. For African-American newborns, there was no further decrease in PRC after 38 weeks suggesting earlier maturation of these fetuses. The study does not propose changing the current 39 weeks threshold for ERCD.

Galactose and its Metabolites Deteriorate Metaphase II Mouse Oocyte Quality and Subsequent Embryo Development by Disrupting the Spindle Structure

Premature ovarian insufficiency (POI) is a frequent long-term complication of classic galactosemia. The majority of women with this disorder develop POI, however rare spontaneous pregnancies have been reported. Here, we evaluate the effect of D-galactose and its metabolites, galactitol and galactose 1-phosphate, on oocyte quality as well as embryo development to elucidate the mechanism through which these compounds mediate oocyte deterioration. Metaphase II mouse oocytes (n = 240), with and without cumulus cells (CCs), were exposed for 4 hours to D-galactose (2 μM), galactitol (11 μM) and galactose 1-phosphate (0.1 mM), (corresponding to plasma concentrations in patients on galactose-restricted diet) and compared to controls. The treated oocytes showed decreased quality as a function of significant enhancement in production of reactive oxygen species (ROS) when compared to controls. The presence of CCs offered no protection, as elevated ROS was accompanied by increased apoptosis of CCs. Our results suggested that D-galactose and its metabolites disturbed the spindle structure and chromosomal alignment, which was associated with significant decline in oocyte cleavage and blastocyst development after in-vitro fertilization. The results provide insight into prevention and treatment strategies that may be used to extend the window of fertility in these patients.

Cyclophosphamide and Acrolein induced oxidative stress leading to deterioration of metaphase II mouse oocyte quality

Abstract Cyclophosphamide (CTX) is a chemotherapeutic agent widely used to treat ovarian, breast, and hematological cancers as well as autoimmune disorders. Such chemotherapy is associated with reproductive failure and premature ovarian insufficiency. The mechanism by which CTX and/or its main metabolite, acrolein, affect female fertility remains unclear, but it is thought to be caused by an overproduction of reactive oxygen species (ROS). Here, we investigated the effect of CTX on metaphase II mouse oocytes obtained from treated animals (120 mg/kg, 24 h of single treatment), and oocytes directly exposed to increasing concentrations of CTX and acrolein (n=480; 0, 5, 10, 25, 50, and 100 &mgr;M) with and without cumulus cells (CCs) for 45 min which correlates to the time of maximum peak plasma concentrations after administration. Oocytes were fixed and subjected to indirect immunofluorescence and were scored based on microtubule spindle structure (MT) and chromosomal alignment (CH). Generation of ROS was evaluated using the Cellular Reactive Oxygen Species Detection Assay Kit. Deterioration of oocyte quality was noted when oocytes were obtained from CTX treated mice along with CTX and acrolein treated oocytes in a dose‐dependent manner as shown by an increase in poor scores. Acrolein had an impact at a significantly lower level as compared to CTX, plateau at 10 &mgr;M versus 50 &mgr;M, respectively. These variation is are associated with the higher amount of ROS generated with acrolein exposure as compared to CTX (p<0.05). Utilization of antioxidant therapy and acrolein scavengers may mitigate the damaging effects of these compounds and help women undergoing such treatment. Graphical abstract Figure. No caption available. HighlightsCyclophosphamide and acrolein treatment deteriorate oocyte quality.The deterioration in oocyte quality progresses in a concentration dependent manner.Oocytes obtained from mice treated with cyclophosphamide displayed similar effect.Cyclophosphamide and acrolein deteriorate oocytes through ROS overproduction.

Missing IUD and utilization of fluoroscopy for management: a case report

INTRODUCTION A missing intrauterine device (IUD) poses challenges in diagnosis and treatment. Extrauterine placement of IUDs may cause difficulty in localization. We report a case where use of fluoroscopy just prior to laparoscopic intervention to remove the missing IUD was beneficial. CASE This case report involves a 35-year-old woman who was diagnosed with a missing thread 2 years after insertion of a levonorgestrel IUD (Mirena). Initial diagnostic workup consisting of abdominal X-ray, an ultrasound and diagnostic laparoscopy failed to localize the IUD. We performed an office pelvic ultrasound which confirmed that an IUD was present but could not specifically localize the IUD in the pelvic or abdominal cavity. Hence, we utilized fluoroscopy just prior to laparoscopy to localize and remove the missing device and allow its removal. CONCLUSION Various imaging modalities aid in detecting a missing IUD, but proper selection of fluoroscopy helped to localize extrauterine placement of an IUD device. We suggest that utilization of this imaging method just prior to laparoscopy may be of value in selected cases.

Imaging and the Infertility Evaluation.

Ultrasound (US) has transformed the fertility evaluation. With 1 consultation, blood work and 1 to 2 USs, the female fertility status can be fully evaluated. The initial US is best done early in the follicular cycle to evaluate the pelvic anatomy and ovarian reserve. A three-dimensional US is important to evaluate for uterine anomalies and color Doppler for any masses. A mid-cycle saline infusion sonohysterogram assesses the endometrial cavity better than a hysterosalpingogram as it identifies the cause of any filling defects. By concurrently adding contrast or agitated saline, tubal patency can be tested. This US-based approach reliably, efficiently, and cost-effectively assesses female infertility.

Dimercapto-1-propanesulfonic acid (DMPS) induces metaphase II mouse oocyte deterioration

In light of the recent lead contamination of the water in Flint, Michigan and its potential adverse outcomes, much research and media attention has turned towards the safety profile of commonly used chelators. Dimercapto-1-propanesulfonic acid (DMPS) typically used in the treatment of lead, mercury and arsenic poisoning also displays a high affinity towards transition metals such as zinc and copper, essential for biological functioning. It is given in series of dosages (0.2-0.4g/day) over a long period, and has the ability to enter cells. In this work, we investigated the mechanism through which increasing concentrations of DMPS alter oocyte quality as judged by changes in microtubule morphology (MT) and chromosomal alignment (CH) of metaphase II mice oocyte. The oocytes were directly exposed to increasing concentration of DMPS (10, 25, 50, 100 and 300μM) for four hours (time of peak plasma concentration after administration) and reactive oxygen species (mainly hydroxyl radical and superoxide) and zinc content were measured. This data showed DMPS plays an important role in deterioration of oocyte quality through a mechanism involving zinc deficiency and enhancement of reactive oxygen species a major contributor to oocyte damage. Our current work, for the first time, demonstrates the possibility of DMPS to negatively impact fertility. This finding can not only help in counseling reproductive age patients undergoing such treatment but also in the development of potential therapies to alleviate oxidative damage and preserve fertility in people receiving heavy metal chelators.

Toxicology in Reproductive Endocrinology

Fertility relies on a series of time dependent events, which are largely regulated by hormones. Concentration and function of these hormones can be altered by exposure of consumed and environmental toxins. It is of important to consider the dose and timing of toxin exposures in assessing the impact of them on reproductive health, however the utility of laboratory assessment in this regard is not well established. General laboratory assessment of reproduction and infertility includes hormone evaluation (hCG, prolactin, TSH, free thyroxin, FSH, LH and androgens among others) and treatment is via various assisted reproductive technologies. Reduction and abstinence from exposure to toxins is recommended to improve reproductive health.