The Use of Atomic Force Microscopy in Comprehensive Assessment of the "Mother-Placenta-Fetus" System in Obstetric and Endocrine Pathology during Pregnancy.

Atomic force microscopy is not very popular in practical health care, therefore, its potential is not studied enough, for example, in obstetrics when studying the "mother-placenta-fetus" system. Our study summarizes the possibilities of using atomic force microscopy for detection of various circulatory disorders and vascular changes at the microscopic level in the uterus (endometrium and myometrium), placenta, and umbilical cord in the main variants of obstetric and endocrine pathology. For instance, in the case of endocrine pathologies, changes in the form of stasis, sludge, diapedesis, ischemia, destruction and separation of endotheliocytes in villous blood vessels were found in the mother. The oxygen content in erythrocytes also naturally decreased in pathologies; poikilo- and anisocytosis were observed.

Delayed villous maturation of the placenta: quantitative assessment in different cohorts.

Placental villous maturation is maximal in the 3rd trimester, with an abundance of terminal villi. Delayed villous maturation (DVM) of the placenta is associated with chromosomal abnormalities, gestational diabetes, and an adverse outcome. This study compares quantitative assessment of vasculo-syncytial membranes (VSM) in cases of liveborn infants, perinatal deaths, and controls. Cases were selected as follows: (1) liveborn infants with a qualitative diagnosis of DVM (n  =  15); (2) controls matched for gestational age whose placentas did not have DVM (n  =  15); (3) stillbirths (SB)/neonatal deaths (NND) showing DVM (n  =  13); and (4) SB from autopsies in which DVM was felt to be the cause of death (COD) (n  =  12). Vasculo-syncytial membranes were counted in 10 terminal villi in each of 10 consecutive high-power fields on 3 slides. Data analysis was carried out using SPSS. Liveborn cases with DVM showed statistically significantly less VSM than controls (mean 1.01 vs 2.42, P < 0.0001). The SB/NND group also showed significantly less VSM than the control group (mean 0.46 vs 2.42, P < 0.0001) and less than the liveborn DVM group (mean 0.46 vs 1.01, P  =  0.001). The COD group was significantly different from the control group (mean 0.42 vs 2.42, P < 0.0001) and the liveborn DVM group (mean 0.42 vs 1.01, P < 0.0001) but not significantly different from the SB/NND group. There is a quantitative reduction in VSM in cases of DVM compared to controls.

[Computer assessment of an apoptotic process dynamics in the nuclei of placental villi syncytiotrophoblast in puerperas after the outburst of herpes virus infection].

The development of an apoptotic process in the nuclei of syncytiotrophoblast was studied in the placenta of 50 puerperas, who had the outburst of herpes virus infection in the second part of their pregnancy (antibody titre--1:12800). Control group was represented by the placental material obtained from 20 women with no disease. Apoptosis was demonstrated in paraffin sections of the material fixed in 10% buffered formalin by in situ labeling of DNA fragments (ISEL-method). The assessment was performed by studying 2000 nuclei from 100 terminal villi in the different areas of the histological section within each placenta. The nuclei in sections were analyzed using a Bio Vision computer program, allowing to detect the areas with the different degree of chromatin condensation during the apoptosis development.

E-cadherin in the assessment of aberrant placental cytotrophoblast turnover in pregnancies complicated by pre-eclampsia.

E-cadherin is a cell-cell adhesion protein expressed in cytotrophoblasts, which is lost as they differentiate and syncytialise. We have exploited E-cadherin as a marker of cytotrophoblasts to investigate villous tissue composition in first and third trimester placentae, both in normal pregnancy and pregnancies complicated by pre-eclampsia. We have achieved this by measuring expression levels of E-cadherin at the mRNA level, using Q-PCR, and at the protein level using semi-quantitative Western blotting. We have also combined E-cadherin immunohistochemistry with morphometric analysis of area measurements to define cytotrophoblast and syncytiotrophoblast compartments. This novel use of E-cadherin has revealed a decrease in the proportion of cytotrophoblasts in villous tissue as pregnancy progresses, in the absence of changes in syncytiotrophoblast cover. Moreover, in pre-eclampsia, placental E-cadherin was raised compared to syncytiotrophoblast, suggesting either exaggerated cytotrophoblast proliferation or impaired cytotrophoblast differentiation, both alterations of potential pathogenic importance.

Stillbirths with placental hemorrhagic endovasculitis: a morphologic assessment with clinical implications.

CONTEXT: Hemorrhagic endovasculitis (HEV) is a vasodisruptive alteration affecting fetal-placental blood vessels of all calibers. Hemorrhagic endovasculitis is found in association with stillbirth and abnormalities of growth and development in livebirths. The role of HEV in the pathogenesis of these conditions is not known. OBJECTIVE: To further understand these events, we compare clinicopathologic features of HEV-affected placentas from stillbirths with those from livebirth pregnancies. Additionally, we assess the relationship of morphologic forms of HEV to clinical events and time of fetal death in utero and evaluate the significance of extensive versus localized HEV lesions in placentas of stillbirths. DESIGN: We reviewed the clinical records and slides from 119 stillbirths with placentas affected by HEV classified above a specified severity level (cases) and 119 matched stillbirths with placentas not affected by HEV (controls). A subset of 21 stillbirth placentas exhibiting focal HEV lesions was similarly evaluated. Slides were graded for HEV, villitis of unknown etiology, chorionic thrombi, villous fibrosis, erythroblastosis, and lesions indicative of maternal hypertension. Hemorrhagic endovasculitis was subcategorized into active, bland, and healed forms and clustered capillary lesions (hemorrhagic villitis). Focal, segmental, and diffuse patterns of villous fibrosis were delineated. Interlesional relationships were established by matching HEV severity indices with severity indices of co-existing lesions. Timing of fetal death was determined by published criteria. Data were analyzed for significance using chi2 and t tests. Results were compared with published analyses of livebirths with placental HEV. RESULTS: Lesions occurring with significant frequency in HEV-affected (case) placentas include villitis of unknown etiology, chorionic thrombi, villous fibrosis, erythroblastosis, and meconium staining. Interlesional relationships were evident between HEV and villous fibrosis, villitis of unknown etiology, and chorionic thrombi. Growth restriction was more common in case versus control infants (P = .02). A segmental pattern of villous fibrosis predominated in cases versus controls and within the case group (P < .001). Time to delivery after fetal death was longer in cases than controls. Active-vasodestructive forms of HEV correlate with shorter intervals of intrauterine retention, whereas bland forms correlate with longer intervals (P = .04). Placentas with focal HEV were associated with coexisting chorionic thrombi and villous fibrosis but not with fetal growth restriction. CONCLUSIONS: Patterns of interlesional interplay are similar in HEV-affected placentas of livebirths and stillbirths. This suggests that the pathogenesis of infant morbidity and mortality is similar in both groups. Active-vasodestructive forms of HEV may precede whereas bland forms may follow intrauterine demise. The segmental pattern of villous fibrosis and high incidences of growth restriction, erythroblastosis, and meconium in cases suggests a chronicity of adverse intrauterine events that may precede fetal loss. Stillbirths with focal HEV lesions are probably not at risk.

[Macroscopic assessment of the placenta using a morphometric grid. Part II: other selected pathologies]. / Ocena makroskopowa lozyska przy zastosowaniu siatki morfometrycznej. Czesc II: Inne wybrane patologie.

Kurt Benirschke has once written that the placenta provides the most accurate records of prenatal history of foetus. Our study included 2498 placentas obtained from pregnancy complicated by pregnancy-induced hypertension (PIH), intrauterine growth retardation (IUGR) and epilepsy. An obstetrician did the planimetric estimation directly after delivery. A measurement of lesions was conducted with using a special own idea's morphometric grid that allowed us to evaluate a placental surface and a relative size of change statement in a very quick and convenient way. Our method is very useful for making a quantitative estimation of pathological changes in the chorionic plate of placenta.

[Macroscopic assessment of the placenta using a morphometric grid. Part III: preterm delivery]. / Ocena makroskopowa lozyska przy zastosowaniu siatki morfometrycznej. Czesc III: Poród przedwczesny.

Kurt Benirschke has once written that the placenta provides the most accurate records of prenatal history of foetus. Our study included 198 placentas obtained from pregnancy complicated by preterm delivery. An obstetrician did the planimetric estimation directly after delivery. A measurement of lesions was conducted with using a special own idea's morphometric gird that allowed us to evaluate a placental surface and a relative size of change statement in a very quick and convenient way. Our method is very useful for making a quantitative estimation of pathological changes in the chorionic plate of placenta.

Alternative method of sampling placentas for the assessment of uteroplacental vasculature.

A new method for the taking and embedding of placental blocks to obtain maximum information regarding the uteroplacental vasculature was developed. This method involves taking en face blocks of the basal plate of the placenta. Results show that normal and abnormal maternal vessels are clearly delineated. In addition to conventional full thickness blocks, which provide information about the chorionic plate such as inflammation and meconium pigment uptake, and about the parenchyma in the other zones remote from the basal plate of the placenta, it is recommended that en face blocks be taken for histological assessment of the placenta.

Why all placentas should be examined by a pathologist in 1990.

Placental pathology is rarely a part of the training for either obstetrician or pathologist. As a result there has been confusion regarding the potential benefits of routine placental examination. These benefits include clarification of the causes of many adverse pregnancy outcomes, improvement of the risk assessment for future pregnancies, and ascertainment of newborn risk for long-term neurodevelopment sequelae. Information on placental abnormalities may reveal the presence of chronic fetal insults and allow their differentiation from acute (peripartum) stresses. Current methods of risk assessment fail to identify the majority of pregnancies that end in prematurity, stillbirth, growth retardation, or fetal distress. We suggest that placental pathology should be a routine component of obstetric-neonatal care.

Diagnostic and therapeutic technology assessment. Chorionic villus sampling.

Chorionic villus sampling has a promising future as a means of early detection of fetal abnormalities. It has widespread application in Europe, and more than 6000 procedures have been performed in the United States. Universal acceptance of the procedure has been delayed because of uncertainties over the true fetal loss rate. Information available today indicates that the fetal loss rate should be in the same range as that for amniocentesis--approximately 1% or less. Confirmation of these estimates awaits release of the data from the large clinical trials currently under way. Modifications of the sampling technique are also under investigation. Transabdominal CVS can also be performed early in pregnancy (six to 15 weeks) with a fine-bore needle and cannula under ultrasonic guidance. It remains to be seen if this offers any advantages or incurs additional risks over transcervical CVS.

Cost-effectiveness ratio of chorionic villi biopsy for the prenatal diagnosis of chromosomal aberrations as compared with amniocentesis.

The observed cost of amniocentesis for the prenatal diagnosis of chromosomal aberrations in pregnancies at risk because of advanced maternal age was compared with the estimated cost of chorionic villi biopsy (CVB). The cost of CVB was estimated to be 22% less than that of amniocentesis when the cost of the sampling procedure, the laboratory charges and the cost of spontaneous and therapeutic abortions were considered.

A practical assessment of ultrasound-guided transcervical aspiration of chorionic villi and subsequent chromosomal analysis.

Aspiration biopsy of trophoblastic villi was performed on 56 patients immediately preceding suction termination of pregnancy, using a malleable metal cannula and ultrasound guidance. Villi were obtained from 47 patients (84%), with sampling success rising to 93% after experience. Immediate complications were noted in 14% of patients and correlated with placental positions situated furthest from the cervical canal. Karyotyping from cultured and/or direct preparations was attempted on the villus samples and was successful in 42. Clarity of the karyotypes obtained from direct preparations in this series was not found to be adequate for diagnostic purposes. A number of practical suggestions which facilitate chorion villus sampling are described.

Assessment of a transcervical aspiration technique for chorionic villus biopsy in the first trimester of pregnancy.

A cannula attached to a syringe was passed through the cervical canal into the uterine cavity in an attempt to aspirate chorionic villi just before termination of a first trimester pregnancy in 82 patients. Chorionic villi were obtained from 40% of these patients. In view of the value of this technique for the first trimester diagnosis of genetic disorder, development of the technique and further trials of transcervical aspiration of chorionic villi would seem warranted.