Solveig M.V. Pflueger

Characterization of Chorioamnionitis in 2nd-Trimester C-Section Placentas and Correlation with Microorganism Recovery from Subamniotic Tissues

Prolonged exposure to infection appears to influence fetal/neonatal development. We characterize the relationship between histologic patterns of inflammation and microorganism recovery from the placentas of live born infants delivered before the 28th postmenstrual week. The subamniotic parenchyma of 835 placentas delivered by cesarean section were cultured and evaluated for specific histologic patterns of inflammation in a blinded fashion. Cases with prolonged membrane rupture were excluded. Microorganisms were recovered from 41% of placentas. Microorganisms found more frequently in placentas with high-grade chorionic plate inflammation include Actinomyces, Prevotella bivia, Corynebacterium sp., Escherichia coli, Peptostreptococcus magnus, multiple species of Streptococci, and Mycoplasma sp., including Ureaplasma urealyticum. These microorganisms were also associated with fetal vasculitis (neutrophilic infiltration of chorionic plate stem vessels or umbilical cord). Recovery of microorganisms from placental parenchyma is associated with histologic inflammation. The same microorganisms responsible for inciting high-grade chorionic plate inflammation are also most likely to promote fetal inflammation.

Histological characteristics of singleton placentas delivered before the 28th week of gestation

Aims: The placenta is a record of the fetal environment and its examination may provide information about the babys subsequent growth and development. We describe the histological characteristics of 947 singleton placentas from infants born between 23 and 27 weeks gestation. Methods: Consent was obtained from mothers who delivered before 28 weeks (clinical estimate). We evaluated the gross and histopathological features of the placenta and assessed pair‐wise correlations between variables. Results: Lesions of uteroplacental circulation (abruption, extensive infarction or thrombosis, marked basal or perivillous fibrin deposition, increased syncytial knots) were inversely related to those associated with inflammation of the membranes and cord. Earlier age favoured inflammatory variables, while older age favoured characteristics attributed to impaired blood flow. We observed inflammation of the chorionic plate in 43%, the cord in 19%, and of chorionic plate vessels in 30%. Of the placentas with umbilical cord inflammation, 8% had no inflammation of the chorionic plate. Conclusions: This study population is unique in its size and recruitment by gestational age rather than birth weight. Inflammation occurred frequently, but not in placentas that had characteristics of vasculopathy. The prevalence of inflammation decreased with increasing gestational age, while vasculopathy increased. Funisitis need not be accompanied by chorionic inflammation.

Cytogenetics of Spontaneous Abortion

Pregnancy loss is quite common, with 15–20% of recognized pregnancies resulting in failure. The majority of these occur early in gestation, although losses in the second and third trimester are not rare. Approximately 2–5% of women will experience two or more losses. The majority of pregnancy failures are associated with cytogenetic abnormalities, with over 50% of early miscarriages and as many as 5% of stillbirths exhibiting abnormal karyotypes.

Recurrent umbilical cord torsion leading to fetal death in 3 subsequent pregnancies: a case report and review of the literature.

During a span of 3.5 years, a 30-year-old, gravida 9, para 3 woman experienced 3 pregnancies complicated by umbilical cord torsion and constriction. In each case, the complication resulted in acute vascular compromise and intrauterine fetal demise. Gross examination disclosed cord constriction and torsion at the fetal end of the cord in each instance. Histologic sections from the cord torsion sites demonstrated fibrosis and deficiencies in Whartons jelly in each case. Cytogenetic studies prepared using fetal villous tissue demonstrated normal karyotypes in fetal cells from the first 2 pregnancies (46,XX and 46,XY, respectively). The karyotype from the third pregnancy showed a 46,XX,del(X)(q24) mutation in 3 of 15 cultured cells, while 12 of 15 cells possessed a normal 46,XX karyotype. This cytogenetic abnormality was not believed to represent the cause of fetal demise in this case. To our knowledge, this is the first report of umbilical cord torsion in 3 pregnancies within one family. The familial clustering observed in this report suggests that a genetic predisposition for umbilical cord torsion may exist in some cases.

Widespread retinal degenerative disease mutation (rdAc) discovered among a large number of popular cat breeds

The recent discovery of a mutational variant in the CEP290 gene (CEP290: IVS50+9T>G), conferring recessive retinal degeneration in Abyssinian and Somali (long-haired Abyssinian) cats (rdAc) prompted a survey among 41 cat breeds (846 individuals) to assess the incidence, frequency and clinical consequence of rdAc. The rdAc allele displayed widespread distribution, observed in 16/43 (37%) breeds, exhibiting a high allele frequency (∼33%) in North American and European Siamese populations. Clinical evaluations demonstrated high concordance between rdAc pathology and the CEP290 (IVS50+9T>G) homozygous genotype (P=1.1E-6), with clinical disease similar to affected Abyssinians/Somalis. This retinal degeneration has not been reported in breeds other than the Abyssinian/Somali and poses a significant health risk particularly in the Siamese breed group. Alertness of the veterinary community and the present availability of commercial diagnostic testing could synergistically enable breeders to reduce the incidence of rdAc blindness in pure-bred cat populations.

Reproduction in a patient with trisomy 8 mosaicism: Case report and literature review

We describe a patient with trisomy 8 mosaicism followed through a sixth pregnancy and discuss issues in phenotypic and genotypic variability, the risk for neoplasia, and reproductive risks.

Triplet placentas: reference values for weights.

The occurrence of twins, triplets, and other multiple births increased significantly between 1970 and 2000 in the United States and other industrialized countries. The number of triplet placentas submitted for examination as pathologic specimens has also markedly increased, but no reference values are published for triplet weights. We examined 196 normal triplet placentas. Specimens with associated conditions known to affect the weights of the placentas were excluded. The gestational ages ranged between 20 and 38 weeks. Mean weights for different gestational ages are summarized as follows: 253 g for 20 weeks, 319 g for 22 weeks, 406 g for 24 weeks, 509 g for 26 weeks, 621 g for 28 weeks, 738 g for 30 weeks, 855 g for 32 weeks, 965 g for 34 weeks, 1065 g for 36 weeks, and 1147 g for 38 weeks. Weight gain of triplet placentas appears to parallel that of twin placentas. The mean values of placental weights for triplets at each gestational age are less than triple those of singleton weights for the same duration of gestation. The placental weights in multiple gestations do not increase proportionately with the number of fetuses.

Establishment of a CYP2C19 Genotyping Assay for Clinical Use

Conversion of clopidogrel (Plavix) to its active metabolite is catalyzed largely by the P450 enzyme 2C19 (CYP2C19). Numerous allelic variants of CYP2C19 exist. The *1 allele is considered wild type, whereas the *2 and *3 alleles have no in vivo enzymatic activity. Conversely, the *17 allele has increased expression, resulting in increased clopidogrel activation. Poor metabolizers (*2/*2 and *2/*3 genotypes) experience higher rates of therapeutic failure. For this reason, we have validated a CYP2C19 genotyping assay for the *1, *2, *3, and *17 alleles. Genomic DNA extracted from 30 deidentified EDTA whole-blood samples from patients was analyzed at 2 independent facilities using specific TaqMan realtime polymerase chain reaction primers and probes. Concordant genotypes were generated on all samples tested. Of the 30 samples, 15 were CYP2C19*1/*1, 8 were CYP2C19*1/*17, 5 were CYP2C19*1/*2, and 2 were CYP2C19*2/*17. There were no CYP2C19*3 alleles or *2/*2 homozygous genotypes detected. This CYP2C19 genotyping assay is appropriate for clinical testing, demonstrating excellent interlaboratory concordance, enabling the selection of the most effective clopidogrel treatment regimen for patients undergoing percutaneous coronary intervention.

The Cytogenetics of Spontaneous Abortion

Pregnancy loss is quite common, with 15–20 % of recognized pregnancies resulting in failure. The majority of these occur early in gestation, though losses in the second and third trimester are not rare. Approximately 2–5 % of women will experience two or more losses. The majority of pregnancy failures are associated with cytogenetic abnormalities, with over 50 % of early miscarriages and as many as 5 % of stillbirths exhibiting abnormal karyotypes.

Using Interphase Fluorescence In Situ Hybridization (I-FISH) to Detect the Transfer of Infant Cells During Breastfeeding

Using fluorochrome-labeled probes for the X and Y chromosomes, fluorescence in situ hybridization (FISH) was used to test for the presence of male cells in the milk of women breastfeeding male babies. After breastfeeding, the women wiped one breast and collected a test sample. Wiping efficiency was evaluated in 2 mothers by running a solution over the nipple after wiping. Cells were collected from both the milk and the rinse samples by centrifugation. The cells were fixed to glass slides for hybridization with the X and Y probes. Two cell populations were observed: small female (XX) cells from the mother, and large epithelial cells that were shown to be male (XY), presumably from the babys mouth. Results varied from 0 to 175 male cells detected per milliliter of breast milk. This finding suggests that small molecules, including possible pathogens, in a babys mouth, might also be passed directly into the breast. J Hum Lact . 24(4):401-405. Usando pruebas con marcadores de fluorocromo para cromosomas X y Y, se utilizó hibridación de fluorescencia in situ (FISH) para detectar la presencia de células masculinas en la leche de madres lactantes de bebes varones. Después de la lactancia, las mujeres se limpiaron un seno y se recolectó la muestra para examinarla. La eficiencia de la limpieza se evaluó en dos madres al lavar con solución el pezón después de la limpieza. Las células se recolectaron de ambas la leche y las muestras lavadas al centrifugarlas. Las células se fijaron en laminillas de vidrio a través de hibridación con las pruebas de X y Y. Se observaron dos poblaciones de células- células pequeñas femeninas (XX) de la madre y células grandes epiteliales que mostraron ser masculinas (XY), presumiblemente de la boca del bebe. Los resultados mostraron un rango de células masculinas de 0-175 detectadas por mililitro de leche materna. Estos hallazgos sugieren que pequenas moléculas, inclusive patógenos, de la boca del bebe pueden también pasar directamente al pecho.