Immunocytochemistry of the placentas of giraffe (Giraffa cameleopardalis giraffa) and okapi (Okapi johnstoni): comparison with other ruminants.

INTRODUCTION: The trophoblast binucleate cell [BNC] is central to the structure and function of all ruminant placentas so far investigated. The Giraffidae are considered to form a separate family within the ruminant suborder. METHODS: The structure and function of two [mid and late pregnant] giraffe placentas and two term okapi placentas have been investigated immunocytochemically. RESULTS: Their major characteristics: polycotyledonary epitheliochorial structure, sequential glucose transport using two transporter isoforms, expression of water transporters in the interplacentomal [IP] and placentomal [P] trophoblast and restriction of calcium transport to the IP trophoblast are similar to those of the ruminant family Bovidae. . Giraffe and okapi also show characteristic ruminant trophoblast binucleate cells (BNC) which migrate and fuse with individual uterine epithelial cells as in the cow. However, there are many fewer BNC, of limited distribution, when compared with other ruminants so far investigated. The giraffe and okapi BNC also show a different range of proteins, Pregnancy Associated Glycoproteins (PAGs) and glycans which clearly distinguish the Giraffidae from the Bovidae. CONCLUSIONS: The results support a separate giraffid family derived from a common ancestry, possessing subpopulations of BNC with potentially different functions.

The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study.

The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins were also apparent. Some differences between Okapi and Giraffe were evident. Most N-linked glycans were similarly expressed in all three species as were fucosyl residues. Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development.

Associations of intrauterine growth restriction with placental pathological factors, maternal factors and fetal factors; clinicopathological findings of 257 Japanese cases.

Intrauterine growth restriction (IUGR) is the leading cause of fetal mortality and morbidity. As an etiology, each of placental findings, maternal factors and fetal factors has been reported to be associated with IUGR, although a comprehensive approach to examine all of these parameters as a cause of IUGR has not been reported. In the present study, therefore, we comprehensively examined the placental findings and maternal and fetal factors in the cases of IUGR (n=257, mean maternal age, 30 years; gestational weeks, 34 weeks) and normal growth pregnancies (n=258, mean maternal age, 30 years; gestational weeks, 33 weeks), and determined risk factors for IUGR. The prevalence of pregnancy hypertension (PHT) (19% vs. 8%, P<0.01), smoking habit (3% vs. 0.7%, P<0.05) and fetal anomaly (3.5% vs. 0.8%, P<0.05) were higher in IUGR cases than normal growth pregnancies. Pathologically, the prevalence of infarction (33% vs. 14%, P<0.05), fetal vessel thrombosis (22% vs. 6%, P<0.001) and chronic villitis (11% vs. 3%, P<0.001) were higher in IUGR cases than those in normal growth pregnancies. A multivariable regression analysis revealed that maternal factors (PHT), fetal factors (anomaly), and placental findings (infarction, fetal vessel thrombosis, and chronic villitis) are independently associated with increased risk of IUGR (all P<0.01).

Role of low placental share in twin-twin transfusion syndrome complicated by intrauterine growth restriction.

OBJECTIVES: Prior studies have demonstrated that donor twin survival following treatment of twin-twin transfusion syndrome (TTTS) was highly associated with donor intrauterine growth restriction (IUGR). Here, we hypothesized that donor IUGR may be attributed in part to low placental share. STUDY DESIGN: The study population consisted of all patients who underwent laser treatment for TTTS at a single institution between 2006-2010. Only those pregnancies with dual survival at birth were included so that placental share information could be interpreted. We examined the relationships between Quintero Stage (with separate analysis of Stage III patients with critically abnormal donor Doppler findings) and low placental share (defined as ≤ 30%) with IUGR (<10th percentile) using chi-square analysis and multivariable logistic regression modeling. RESULTS: Of 210 patients treated, 159 (75.7%) had dual survivors at birth. Of these, placental share was documented in 90 cases (56.6%). Twenty-seven (30.0%) had low placental share, and 37 (41.1%) had IUGR. IUGR was associated with low placental share (63.0% vs. 31.7%, P = 0.0116). IUGR was also associated with Stage III patients (57.4% vs. 23.3%, P = 0.0021), and in particular with Stage III patients with donor involvement (77.8% vs. 25.4%, P < 0.0001). In logistic regression modeling, both low placental share and Stage III with donor involvement were independent risk factors for IUGR (OR = 3.5 [1.2-10.3], P = 0.0206, and OR = 10.1 [3.3-30.6], P < 0.0001, respectively). CONCLUSIONS: Donor IUGR in TTTS pregnancies appears to be associated, in part, with low placental share.

Residual vascular communications in twin-twin transfusion syndrome treated with sequential laser surgery: frequency and clinical implications.

OBJECTIVES: The goal of fetoscopic laser surgery for twin-twin transfusion syndrome (TTTS) is to ablate all placental vascular communications, thereby separating the fetal circulatory systems. We sought to ascertain the frequency and clinical implications of residual vascular communications (RVC) post preferential sequential selective laser photocoagulation of communicating vessels (SQLPCV). STUDY DESIGN: TTTS placentas treated via preferential SQLPCV were examined. Patency of vascular communications was assessed via water and/or milk injections. Cases with intrauterine fetal demise or placental disruption were excluded. Outcomes with and without RVC were compared. RESULTS: One hundred seventy-four TTTS patients were treated during the study period. Dual survival at birth was 76% (133/174). Of the 133 dual survivors, 105 (79%) submitted an intact placenta. Five of these 105 placentas had RVC (4.8%). Comparison of RVC versus non-RVC cases revealed the following: gestational age at delivery 28.7(6.5) vs. 33.4(3.3) weeks (p=0.178); recipient birth weight 1287(1061) vs. 1973(610) grams (p=0.020); donor birth weight 1429(1369) vs. 1653(715) grams (p=0.518); donor central/eccentric placental cord insertion 80% vs. 17% (p=0.006). One case required a second laser surgery to complete the laser ablation; this placenta did not have RVC after delivery. Otherwise there were no cases of persistent TTTS. One of the 5 RVC cases (20%) exhibited neonatal findings consistent with twin anemia-polycythemia sequence (TAPS), while none of the non-RVC cases had TAPS (p=0.005). CONCLUSIONS: The rate of RVC was less than 5% among gestations with dual survivors post preferential SQLPCV treatment for TTTS.

The monozygotic twinning process, the twin-twin transfusion syndrome and acardiac twins.

This paper suggests that during the time of the "splitting process" that produces monozygotic twins the number of cells may not be evenly distributed to the future twins. The outcome of this 'irregular division' may be the twin-to-twin transfusion syndrome and acardiac twins.

Placental tumor from an East African bongo (Eurycerus isaaci) fetus.

This case report describes a placental tumor from a stillborn East African bongo fetus. On gross examination, the tumor was reminiscent of a chorangioma; however, based on histomorphology, histochemical and immunophenotypic characterization, comparison to 21 human cases of chorangioma, and embryogenesis, this tumor is hypothesized to be a placental stem cell tumor. Histologically, the tumor had 2, often commingled, morphologies with one area having dense cellularity and scant vascularity and a second area having scant cellularity and prominent vascularity. Pleomorphic mesenchymal cells were arranged in intersecting fascicles, often whorled around, or were contiguous with vascular channels and had features of skeletal muscle, smooth muscle, pericyte, endothelial, fibrous, and undifferentiated mesenchymal cells. Phosphotungstic acid-hematoxylin outlined cross-striations in some cells. Cells variably were immunoreactive for vimentin, CD31, von Willebrand factor VIII, desmin, muscle specific actin, alpha-smooth muscle actin, and skeletal myosin and negative for cytokeratin antigens. The relationship of this tumor to the presence of fetal musculoskeletal and omental anomalies is discussed.

Effect of porcine zonae pellucidae immunisation on ovarian follicular development and endocrine function in domestic ewes (Ovis aries).

Domestic ewes (Ovis aries) were immunised with porcine zonae pellucidae (pZP) or pZP conjugated to keyhole limpet haemocyanin (KLH) in adjuvant(s) to examine the feasibility of the species to serve as a model for further development of pZP-based vaccines in ungulates. Two immunisation groups were employed, with a third group receiving only adjuvant (n = 5 per group). Early in the study, oestrous activity was monitored by the use of a vasectomised ram fitted with a marking harness. Eventually, ewes were exposed to an intact ram for breeding. In addition, weekly serum and every-other-day faecal samples were collected to measure pZP antibodies and progesterone metabolite concentrations respectively. At the conclusion of the study, fecundity was established, and ovarian tissue was examined. Ewes immunised against pZP : KLH with adjuvant produced minimal antibody absorbance levels, displayed normal oestrous cycles, became pregnant upon introduction of the intact ram and exhibited normal ovarian histopathology. Ewes immunised against pZP with adjuvant produced high antibody absorbance levels, were acyclic following primary immunisation and were infertile. Examination of the ovarian tissue revealed atrophic changes that included: (1) the absence of growing follicles; (2) significant reduction in the number of primordial follicles; and (3) the presence of abnormal granulosa cell clusters lacking oocytes. Antisera displayed immunoreactivity to the major components of pZP, and immunohistochemical labelling of ovarian tissue showed specificity to the ZP. These data are the first generated in an ungulate species showing deleterious effects of pZP immunisation on folliculogenesis and oestrous cyclicity.

Umbilical arterial thrombosis with vascular wall necrosis: clinicopathologic findings of 11 cases.

Thrombosis of umbilical cord vessels is associated with a high perinatal mortality. Most cases are venous thrombi, arterial thrombi are uncommon. To clarify clinical and pathological features of this entity, we reviewed 11 cases with umbilical artery thrombosis. The gestational ages were 33-40 (mean 36.8) weeks. Three cases (38%) were associated with severe intrauterine growth retardation and two cases (25%) led to intrauterine fetal deaths. All cases had completely occlusive thrombi of one umbilical artery. The arteries with thrombosis demonstrated partial necrosis of vascular wall. The pathogenesis is unclear, but nine cases (82%) had cord abnormalities (long cord, peripheral cord insertion, short cord with twist, funisitis). This paper is the first report of clinical and pathological features of umbilical artery thrombosis. The results of this study confirm the clinical importance and unique histological findings of umbilical artery thrombosis. The recognition of umbilical artery thrombosis is necessary to establish the diagnosis and treatment of this condition.

Funisitis associated with leptospiral abortion in an equine placenta.

Funisitis, inflammation of the umbilical cord, is well recognized in human placentas. This report describes a case of funisitis associated with leptospiral infection in the placenta of a Thoroughbred foal born prematurely. The umbilical cord had diffuse superficial yellow discoloration along its entire length. Microscopic evaluation showed an exudate of neutrophils admixed with fibrin on the surface. Warthin-Starry staining showed spirochetes in the Wharton's jelly of the umbilical cord. A locally extensive, severe placentitis not involving the star and allantoic cystic hyperplasia were the other lesions observed in the allantochorion. Leptospira funisitis is similar to the funisitis of congenital syphilis in humans, although there are some major microscopic differences. In Leptospira funisitis, lesions were limited to the cord surface, whereas in lesions in human umbilical cords with Treponema pallidum infection, the changes are observed mostly around the vessels and in the Wharton's jelly.

Ovarian choriocarcinoma in a rhesus monkey associated with elevated serum chorionic gonadotropin levels.

A clinically normal, 3-year-old female rhesus monkey (Macaca mulatta), which was part of a routine toxicology study, had a mass in the right ovary with metastases to the adjacent mesentery and lungs. The histologic features and immunohistochemistry results were consistent with the diagnosis of choriocarcinoma. Neoplastic cell types included cytotrophoblast (positive for cytokeratin), syncytiotrophoblast (positive for human chorionic gonadotropin), and extravillous trophoblast (positive for human placental lactogen). Because the neoplasm was present in the ovary, the uterus was normal, and the animal was not currently pregnant, this was considered a primary ovarian neoplasm of germ cell origin. The monkey had elevated serum levels of chorionic gonadotropin at the beginning of the study, indicating that, as in women, choriocarcinomas in monkeys can be associated with increased gonadotropin levels and that the tumor was preexisting at the start of the toxicology study.

Equine placental teratoma.

The placenta from a 300-day-gestational age, female, Arabian equine fetus was examined. Multifocal to coalescing, 0.5- to 4-cm-diameter, white, smooth nodules covered 50% of the placenta. Microscopic evaluation of the nodules revealed undifferentiated germ cells and a haphazard arrangement of immature, mesenchymal stroma, cartilage, squamous cornifying epithelium, scattered ducts and secretory acini lined by cuboidal to columnar epithelium, and mineralized foci. No umbilicus, arrangement about an axial skeleton, or organized polarity of structures was present. The lesion was diagnosed as a placental teratoma, a lesion not reported in species other than man.

Comparative cytogenetics of the African elephant (Loxodonta africana) and Asiatic elephant (Elephas maximus).

G- and C-banded karyotypes of the two extant species of the mammalian order Proboscidea are presented for the first time. Chromosome complements were 2n = 56 in both Loxodonta africana and Elephas maximus. Comparisons between the species demonstrated a high level of chromosome band homology, with 26 conserved autosomal pairs. The normal diploid karyotype of L. africana had 25 acrocentric/telocentric and two metacentric/submetacentric autosomal pairs. E. maximus differed by having one less acrocentric and one additional submetacentric pair due to either a heterochromatic arm addition or deletion involving autosomal pair 27. Several acrocentric autosomes of L. africana exhibited small short arms that were absent in homologous chromosomes of E. maximus. The X chromosomes in both species were large submetacentric elements and were homologous. However, the small acrocentric Y chromosomes differed; in E. maximus it was slightly larger and had more distinct G-bands than its counterpart in L. africana. Extant Elephantidae appear to be relatively conservative in their rates of chromosomal change compared to some other mammalian families. The high-quality banded karyotypes presented here should prove useful as references in future chromosome analyses of elephant populations and in comparative cytogenetic studies with other ungulate orders.

Exposure to the dioxin 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces squamous metaplasia in the endocervix of cynomolgus macaques.

Female cynomolgus macaques (n = 11) were treated orally with graded doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Cervical tissue was recovered at necropsy 1.2-2.7 years later and examined using routine histopathology. Results were compared histologically with cervical tissue from untreated, age- and parity-matched controls. Significant squamous epithelial metaplasia was observed in the endocervix of 9 of 11 TCDD-treated animals, and the degree of severity was dose dependent. In contrast, minimal or no pathological changes were observed in eight of nine control animals and one animal had only mild squamous metaplasia. These results suggest that TCDD exposure induces epithelial transdifferentiation in the primate cervix. Consequently, the TCDD-treated macaque may serve as a predictable animal model for the study of cervical epithelial transdifferentiation and for examining the relationship between squamous metaplasia and cervical oncogenesis both at the cellular and at the molecular level.

Partial caudal duplication in a newborn associated with meningomyelocele and complex heart anomaly.

BACKGROUND: Caudal duplication is a spectrum of rare congenital anomalies with a possible heterogeneous pathogenesis including incomplete separation of monovular twins. METHODS: We report an autopsy case of a full-term infant with incomplete caudal duplication syndrome associated with multiple anomalies. RESULTS: These anomalies included a duplicated penis; double urinary bladder with an attenuated tunica muscularis; duplication of lower bowel with two ilia, appendices and colons; colonic hypogangliosis and left imperforated anus associated with rectourethral fistula. Other anomalies consisted of sacral meningomyelocele, sacral duplication with hypoplastic left sacrum and pelvic bones, muscle atrophy and hypoplasia of the left lower extremity, abnormal lobation of liver with stomach entrapment, omphalocele, and right atrial isomerism syndrome. The complex pattern of anomalies suggests the possibility that partial caudal duplication might be part of the spectrum of conjoined twinning.

Morbidity, mortality, and placental pathology in excessively long umbilical cords: retrospective study.

The purpose of this study was to compare specific fetal, maternal, and placental factors, including neonatal morbidity and mortality, in infants with umbilical cords (UCs) of normal length to the same factors in infants with excessively long umbilical cords (ELUCs). We performed an 18-year retrospective chart review of the medical records of mothers and infants with ELUCs (926 cases) and normal-length UCs (200 cases) and recorded maternal factors, fetal factors, and neonatal outcomes. Corresponding placental pathologic reports and slides were reviewed. Statistical analysis comparing the two groups included univariate and multivariate analyses. ELUCs were significantly associated with certain maternal factors (systemic diseases, delivery complications, increased maternal age), fetal factors (non-reassuring fetal status, respiratory distress, vertex presentation, cord entanglement, fetal anomalies, male sex, increased birth weight), gross placental features (increased placental weight, right-twisted cords, markedly twisted cords, true knots, congestion), and microscopic placental features (nucleated red blood cells, chorangiosis, vascular thrombi, vascular cushions, meconium, increased syncytial knots, single umbilical artery). Some of these histopathologic features have previously been associated with fetal hypoxia and/or altered blood flow in the placenta. Infants with ELUCs were found to be at a significantly increased risk of brain imaging abnormalities and/or abnormal neurological follow-up. In addition, mothers with a history of an ELUC are at increased risk of a second long cord.

Trisomy 17 in a bonobo (Pan paniscus) and deletion of 3q in a lowland gorilla (Gorilla gorilla gorilla): comparison with human trisomy 18 and human deletion 4q syndrome.

A female bonobo (Pan paniscus) born at the San Diego Zoo exhibited inability to nurse and progressive weakness plus multiple congenital abnormalities including aural canal atresia and stenosis, malformed auricles, clenched hands, lordosis, agenesis of the caudal vertebra and cardiac abnormalities. Chromosome analysis identified the bonobo as being trisomic for chromosome 17, the homolog of human chromosome 18. Genotyping with human microsatellites suggested the extra chromosome was maternal in origin. In addition, a male lowland gorilla (Gorilla gorilla gorilla), also born at the zoo, exhibited postnatal growth retardation, facial dysmorphisms and small hands with short fingers. Karyotype analysis revealed the gorilla carried a deletion of the distal q arm of chromosome 3, the homolog of human chromosome 4. The phenotypic and karyotypic abnormalities found in the bonobo and gorilla were consistent with the characteristics of human trisomy 18 and human deletion 4q syndrome, respectively.

The placenta in multiple pregnancy: outstanding issues.

The present paper reviews the placental development in multiple gestations and highlights the twin-to-twin transfusion syndrome, its recognition and possible therapy, and some of the consequences of fetal death of one twin. It presents data on discordance for structural anomalies in monozygotic twins. These aspects are discussed with respect to their possible pathogenetic origins.