Maturation Of Placenta Research Articles

Combining RNAscope, Immunohistochemistry (IHC) and Digital Image Analysis to Assess Podoplanin (PDPN) Protein and PDPN_mRNA Expression on Formalin-Fixed Paraffin-Embedded Normal Human Placenta Tissues.

The expression and function of podoplanin (PDPN) in the normal human placenta has been debated in placental evaluation. This study emphasizes the importance of a multimodal approach of PDPN expression in normal human placentas. A complete examination is performed using immunohistochemistry, RNAscope and automated Digital Image examination (DIA) interpretation. QuPath DIA-based analysis automatically generated the stromal and histological scores of PDPN expression for immunohistochemistry and RNAscope stains. The umbilical cord's isolated fibroblasts and luminal structures expressed PDPN protein and PDPN_mRNA. RNAscope detected PDPN_mRNA upregulation in syncytial placental knots trophoblastic cells, but immunohistochemistry did not certify this at the protein level. The study found a significant correlation between the IHC and RNAscope H-Score (p = 0.033) and Allred Score (p = 0.05). A successful multimodal strategy for PDPN assessment in human placentas confirmed PDPN expression heterogeneity in the full-term human normal placenta and umbilical cord at the protein and mRNA level. In placental syncytial knots trophoblastic cells, PDPN showed mRNA overexpression, suggesting a potential role in placenta maturation.

Tribrachius parapagus diencephalus conjoined twin

A conjoined twin of male gender, weighing 3 kgs was born at 36 weeks of gestation to a primigravida mother. The twin cried immediately after birth. The twin had a single trunk, three upper limbs, two lower limbs and two separate heads joined at the trunk , single anal opening and male external genitalia. Antenatal USG at 35 weeks showed polycephaly live intrauterine fetus with breech presentation with anteriorly placed placenta maturity grade II with gross fetal anomaly, polyhydramnios with distended fetal urinary bladder (bladder volume: 63ml). The twin had respiratory distress on admission, so oxygen support was given. Babygram done was suggestive of two separate vertebral columns from cervical to lumbosacral region. 2D echo shows 2 hearts. Twin was managed conservatively, started on NG feeds and discharged in stable condition.

The X-linked splicing regulator MBNL3 has been co-opted to restrict placental growth in eutherians

Understanding the regulatory interactions that control gene expression during the development of novel tissues is a key goal of evolutionary developmental biology. Here, we show that Mbnl3 has undergone a striking process of evolutionary specialization in eutherian mammals resulting in the emergence of a novel placental function for the gene. Mbnl3 belongs to a family of RNA-binding proteins whose members regulate multiple aspects of RNA metabolism. We find that, in eutherians, while both Mbnl3 and its paralog Mbnl2 are strongly expressed in placenta, Mbnl3 expression has been lost from nonplacental tissues in association with the evolution of a novel promoter. Moreover, Mbnl3 has undergone accelerated protein sequence evolution leading to changes in its RNA-binding specificities and cellular localization. While Mbnl2 and Mbnl3 share partially redundant roles in regulating alternative splicing, polyadenylation site usage and, in turn, placenta maturation, Mbnl3 has also acquired novel biological functions. Specifically, Mbnl3 knockout (M3KO) alone results in increased placental growth associated with higher Myc expression. Furthermore, Mbnl3 loss increases fetal resource allocation during limiting conditions, suggesting that location of Mbnl3 on the X chromosome has led to its role in limiting placental growth, favoring the maternal side of the parental genetic conflict.

Study of placental maturity by ultrasound in pregnancy with sickle cell disease/trait & correlation with normal pregnancy

Aims and Objectives: The aim of the study is to evaluate the effect of sickle cell disease/trait on the maturation of placenta and further correlation with maturation of placenta in normal individuals.Material and Methods: Antenatal ultrasound of 45 pregnant patients from each three different categories (i.e., sickle cell disease; sickle cell trait and normal) was done. Sickle cell disease (SCD) and Sickle cell trait (SCT) patients were diagnosed on the basis of serum electrophoresis test. The degree of placental maturation, thickness and location of the placenta evaluated. The data collected from the subjects then further compared for any difference between the degree of maturity between three study groups. Results: The correlation between placental grading and gestational age in all three categories was not statistically significant (P value > 0.05). Conclusion: There is no significant difference noted among all the three categories in term of placental maturation and fetal weight measures.

Third Trimester Placental Grading by Ultrasonography and its Relationship with Fetal Outcome

Background: Antenatal ultrasound assessment of placental morphology plays an important role in evaluating fetal health, revealing abnormalities, such as infract & calcification. Abnormal placental maturity is associated with poor pregnancy outcome. In some high risk pregnancy placental maturity is accelerated as in pregancy –induced hypertension (PIH), intrauterine growth retardation (IUGR), whether in other high risk cases like diabetes and Rh-isoimmunization there is delayed placental maturation.
 Objectives: This was a cross sectional type of descriptive study. The study was carried out to assess placental grading by USG in high risk and normal pregnancy for predicting neonatal outcome.
 Material and method: The study was carried out in the Department of Obstetrics & Gynaecology in collaboration with the Department of Radiology & Imaging of Rajshahi Medical College Hospital from January 2012 to December 2013. : A total 200 pregnant mother, attending the inpatient department constituted study population who delivered their babies at Rajshahi Medical College Hospital were selected in antenatal period for USG examination to detect placental grade.
 Result: Ultrasonography showed, 35% had Grade-II placental maturity, 33.5%- Grade-III, 25% Grade - I and 6.5% had Grade-0 maturity. Majority of the mother (75%) were normal and had healthy baby. The relationship of placental maturity with gestational age of mother in normal pregnancy and in high risk pregnancy was found to be statistically significant. Placental maturity and fetal outcome in normal pregnancy was found significantly associated (p<0.001) but in high risk pregnancy it was not associated significantly (p>0.5). Early maturation of placenta in hypertensive women and delayed maturation in diabetic women were observed.
 Conclusion: Sonographic diagnosis of grade-III placenta has been reported to be as excellent predictor of fetal lung maturity than difficult and hazardous invasive procedure like amniocentesis.
 TAJ 2020; 33(2): 94-99

Cigarette smoke induced neural tube defects by down-regulating noggin expression.

Maternal smoking or passive smoking during gestation increases the risk of congenital birth defects, including neural tube defects (NTDs). Cigarette smoke is a recognized teratogen that causes NTDs, although the mechanisms are not well understood. An established model of passive smoking in pregnant golden hamsters was used to observe the effect of cigarette smoke on neural tube development using scanning electron microscopy. Level of noggin expression in placenta and neural tube was evaluated by immunohistochemistry. Our results indicated that cigarette smoke can cause the neural tube closure of the golden hamster embryo to be delayed or not closed. In normal placental tissues noggin protein and RNA levels were highly expressed, and the expression level in the term placenta was lower than in that of the first trimester or second trimester through analyzing the Human Protein Atlas and the Gene Expression Omnibus (GEO) databases. Cigarette smoke can down-regulate noggin expression in the placenta and promote cell apoptosis. We observed that noggin expression was reduced and BMP2 expression was increased by cigarette smoke. Cigarette smoke may cause cell apoptosis and accelerate placenta maturation. Meanwhile, cigarette smoke may inhibit the development of the embryo, and lead to the formation of NTDs via downregulating the expression of noggin and dis-inhibition of BMP2.

Identification of suspicious invasive placentation based on clinical MRI data using textural features and automated machine learning.

The aim of this study was to investigate whether intraplacental texture features from routine placental MRI can objectively and accurately predict invasive placentation. This retrospective study includes 99 pregnant women with pathologically confirmed placental invasion and 56 pregnant women with simple placenta previa. All participants underwent magnetic resonance imaging after 24 gestational weeks. The placenta was segmented in sagittal images from both turbo spin echo (TSE) and balanced turbo field echo (bTFE) sequences. Textural features were extracted from the both original and Laplacian of Gaussian (LoG)-filtered MRI images. An automated machine learning algorithm was applied to the extracted feature sets to obtain the optimal preprocessing steps, classification algorithm, and corresponding hyper-parameters. A gradient boosting classifier using all textual features from original and LoG-filtered TSE images and bTFE images identified by the automated machine learning algorithm achieved the optimal performance with sensitivity, specificity, accuracy, and area under ROC curve (AUC) of 100%, 88.5%, 95.2%, and 0.98 in the prediction of placental invasion. In addition, textural features that contributed to the prediction of placental invasion differ from the features significantly affected by normal placenta maturation. Quantifying intraplacental heterogeneity using LoG filtration and texture analysis highlights the different heterogeneous appearance caused by abnormal placentation relative to normal maturation. The predictive model derived from automated machine learning yielded good performance, indicating the proposed radiomic analysis pipeline can accurately predict placental invasion and facilitate clinical decision-making for pregnant women with suspicious placental invasion. • The intraplacental texture features have high efficiency in prediction of invasive placentation after 24 gestational weeks. • The features with dominated predictive power did not overlap with the features significantly affected by gestational age.

Changes in the function of the fetoplacental complex under the influence of the hepatitis B virus

The leading clinical syndrome in pregnant women with HBV-infection is placental dysfunction, which is due to morphological and functional changes in the placenta and manifests itself as a violation of the state of the fetus and its development. The objective: of the study was to eliminate perinatal and obstetric complications in pregnant women with HBV-infection based on the detection of echoscopic deviations of the fetoplacental complex. Materials and methods. 228 women were examined in the period of 30–34 weeks of gestation: the basic group (146 pregnant women with HBV-infection) and the control group (82 pregnant women without HBV-infection). Results. The most common symptom of ultrasound in pregnant women with hepatitis B was thickening myometrium, as changes in the thickness and structure of the placenta. It is necessary to note the presence of low attachment of the placenta in women with HBV-infection. In women of the basic group, asymmetric form of the fetal growth retardation was diagnosed in 21.23±3.38% women of the basic group, subcompensated respiratory movements – 34.93±3.95% and subcompensated fetal activity – 41.10±4.07%, as well as a violation of fetal tones – 30,14±3,80%, with a reliability of p<0.05 compared with the control group. In the presence of HBV-infection, ultrasound changes were particularly demonstrative from such indicators as premature placenta maturation to grade 1 – in 37.57±4.01%; myometrium pathology in the placental zone – 44.52±4.11%; the combination of thinning or thickening with premature ripening – 47.26±4.13%; polyhydramnios – 52.74±4.13% and oligohydramnios – 25.34±3.60% (p<0.05). Conclusion. Thus, placental dysfunction in pregnant women with HBV-infection develops under the influence of various factors, the leading of which is the virus of hepatitis B. Key words: hepatitis B, HBV-infection, pregnancy, placenta.

Premature “aging” of placenta: myth or reality?

There was performed a retrospective analysis of pregnancy outcomes in 585 women with different placental maturity. There is given an assessment of premature “aging” of placenta in the prognosis of fetal and newborn development. The third degree of maturity of placenta at 32–34 weeks of gestation was observed more often with normal body weight of children at birth than with slowing fetal growth. In 62.5 % of cases of placental degree registration at 32–34 weeks were born newborns weighing 4000 g. The results suggest that the 3rd degree of placental maturity less than 36 weeks doesn't have a significant impact on the condition and development of the fetus and newborn.

Geranylgeranyl pyrophosphate synthase facilitates the organization of cardiomyocytes during mid-gestation through modulating protein geranylgeranylation in mouse heart.

With the maturation of placenta, ventricular chamber maturation enhances cardiac contractile performance to adapt to the metabolic demand of growing embryo. The organization of cardiomyocytes is required for the morphological remodelling in ventricular chamber maturation. However, the mechanism governing the establishment of cardiac cytoarchitecture during ventricular chamber maturation is still poorly studied. Here, we found that the expression of geranylgeranyl pyrophosphate synthase (Ggpps), which mediates protein geranylgeranylation, increased in the mouse heart after the onset of placental function. By using different Cre lines, we found that the cardiac inactivation of Ggpps by the Nkx2.5Cre/+ line disrupted protein geranylgeranylation as early as E9.5, which affected ventricular chamber maturation and resulted in mid-gestational embryonic lethality. In contrast, α-SMA-Cre line mediated the disruption of protein geranylgeranylation from E13.5 did not affect embryonic heart development. Further analysis of Nkx2.5Cre/+; Ggppsfl/fl mutants showed that the loss of Ggpps caused disorganized cardiac cytoarchitecture as early as E11.5 by disturbing cell-cell junctions. Ggpps inactivation decreased Rho GTPase geranylgeranylation and their activity, which might account for the disruption of cell-cell junctions. Moreover, elevating the protein geranylgeranylation by supplement of geranylgeranyl pyrophosphate (GGPP) could recover the Ggpps deficient induced defects of cytoarchitecture and cell-cell junctions in vitro and in vivo. Our present study demonstrates that GGPPS-mediated protein geranylgeranylation plays an indispensable role in the ventricular chamber maturation and acts as a stage-specific signal to regulate the establishment of cardiac cytoarchitecture during mid-gestation.

Placental insufficiency among high-risk pregnancies with a normal umbilical artery resistance index after 32 weeks

ObjectiveTo determine the incidence of abnormal multi-vessel Doppler values among advanced pregnancies at risk of suboptimal placentation but with a normal umbilical artery resistance index (RI), and to assess whether clinical and ultrasonography findings can identify them. MethodsIn a prospective cross-sectional study at Tygerberg Hospital, South Africa, women with high-risk pregnancies but normal umbilical artery RI after 32weeks underwent ultrasonography (fetal biometry, liquor, and placenta maturation) and Doppler assessment (uterine, umbilical, and middle cerebral arteries) between February 11 and October 21, 2013. Study data were compared among four groups: fetuses with normal uterofetoplacental Doppler values and those with any abnormal pulsatility index, each subdivided into small for gestational age (SGA) and appropriate for gestational age (AGA) by estimated fetal weight. ResultsOf 210 participants, 72 (36.2%) had abnormal Doppler results, and 60 (28.6%) fetuses were SGA (38 [63.3%] with abnormal Doppler results). Clinical characteristics did not differ between groups with normal or abnormal Doppler values; however, among normal Doppler results, SGA pregnancies demonstrated poorer fundal growth (P=0.006). Significant associations existed between abnormal Doppler results and asymmetric growth, inappropriately advanced placental maturation, and reduced liquor volume (all P≤0.04), but with very low sensitivities (3.9%, 4.8%, and 14.5%, respectively). ConclusionMaternal characteristics and imaging variables did not reliably identify more than one-third of pregnancies with evidence of suboptimal placentation.

Discriminative Learning for Automatic Staging of Placental Maturity via Multi-layer Fisher Vector.

Currently, placental maturity is performed using subjective evaluation, which can be unreliable as it is highly dependent on the observations and experiences of clinicians. To address this problem, this paper proposes a method to automatically stage placenta maturity from B-mode ultrasound (US) images based on dense sampling and novel feature descriptors. Specifically, our proposed method first densely extracts features with a regular grid based on dense sampling instead of a few unreliable interest points. Followed by, these features are clustered using generative Gaussian mixture model (GMM) to obtain high order statistics of the features. The clustering representatives (i.e., cluster means) are encoded by Fisher vector (FV) for staging accuracy enhancement. Differing from the previous studies, a multi-layer FV is investigated to exploit the spatial information rather than the single layer FV. Experimental results show that the proposed method with the dense FV has achieved an area under the receiver of characteristics (AUC) of 96.77%, sensitivity and specificity of 98.04% and 93.75% for the placental maturity staging, respectively. Our experimental results also demonstrate that the dense feature outperforms the traditional sparse feature for placental maturity staging.

Automatic grading of placental maturity based on LIOP and fisher vector.

Currently, the evaluation of placental maturity has mainly focused on subjective measure, which highly depends on the observation and experiences of the clinicians and not reliable. This paper proposes a new method for grading placenta maturity in B-mod ultrasound (US) images automatically based on local intensity order pattern (LIOP) and fisher vector (FV). After extracting invariant LIOP feature from the affine covariant region, the feature is encoded by FV to improve the classification accuracy and reduce the processing time. Experimental results show the effectiveness of the proposed method with an accuracy of 0.9375, a sensitivity of 0.9804 and a specificity of 0.9375 for the placental maturity grading. Moreover, experimental results demonstrate that the LIOP feature outperforms the traditional SIFT feature for grading.

Immunoregulation by Cytolytic Pathways, Mucins and Progesterone at the Maternal-Fetal Interface

The maternal-fetal interface of pregnant mammals is characterized by a sensitive balance between hormones, cytokines, humoral factors and cellular interactions. Progesterone activity leads to the transformation of endometrial cells into the decidual phenotype and ensures the integrity of the maternal-fetal interface during trophoblast invasion and placenta maturation. Communication between the genomic and non-genomic progesterone-regulated signaling pathways could be of critical importance for the establishment of a correct endocrine-immune interaction in the human endometrium during the establishment and maintenance of pregnancy. Cytolytic cells (NK and T lymphocytes) infiltrating the decidua of pregnancy are heavily equipped with cytolytic molecules perofrin and granulysin. These molecules have very important role(s) in the maternal tolerance of fetoplacental unit, control of trophoblast invasion, defense against infective agents and malignancies, as well as immunomodulatory functions. Mucins MUC- 1 and TAG-72 are important features of secretory phase decidual reaction, while their dissapearance from the surface of the epithelial cells enables apposition, adhesion and infiltration of high quality blastocysts at the precise area at the time of the implantation. Muc-1 and TAG-72 induce antiinflammatory orientation of the decidual antigen presenting cells.

Amphiregulin regulates the production of human chorionic gonadotropin in trophoblasts

Aims The aim of this study was to investigate the significance of epidermal growth factor receptor (EGFR) ligands produced in syncytiotrophoblasts during normal pregnancy. Main methods We examined the expression of EGFR ligands in human pregnancy by real-time PCR, and analyzed the relationship between EGFR ligands and human chorionic gonadotropin (hCG) or human placental lactogen in amniotic fluid by ELISA. In addition, we also examined the EGFR ligands in syncytiotrophoblasts and the amount of hCG secretion in JAR, JEG3 and BeWo cells in the presence of each EGFR ligand. Key findings In order to identify possible candidates among the EGFR ligands, we examined the predominant expression of an EGFR ligand in the chorionic villi and amniotic fluid during normal pregnancy, and analyzed the relationship between EGFR ligands and hCG in trophoblastic model cells. Amphiregulin was primarily expressed throughout human pregnancy and stimulated the secretion of hCG, indicating that amphiregulin is a key molecule among EGFR ligands. Significance Amphiregulin may play a pivotal role in the development or maturation of placenta.

ORIGINAL ARTICLE: Genetic Predisposition to Idiopathic Recurrent Spontaneous Abortion: Contribution of Genetic Variations in IGF‐2 and H19 Imprinted Genes

Problem Recurrent spontaneous abortion (RSA) is a common clinical problem with a complex etiology of genetic and non‐genetic causes, which remains to be fully determined. IGF‐2 stimulates trophoblast invasion, proliferation and maturation of placenta, while H19 RNA suppresses growth. As genomic imprinting plays a critical role in the development of placenta and embryo, our aim was to evaluate the possible role of variations in IGF‐2 and H19 imprinted genes as factors of predisposition for RSA.Method of study A case–control study was conducted to determine the association between IGF‐2 and H19 gene polymorphisms and the susceptibility to RSA in 113 couples with RSA and 226 controls. PCR/RFLP were performed to analyze IGF‐2 ApaI and H19 HhaI polymorphisms.Results We found a statistically significant difference in the genotype frequency distribution of IGF‐2 ApaI polymorphism between males from couples with RSA and healthy males (χ2(2) = 45.12; P < 0.0001). There were no differences in the genotype and allele distribution of H19 polymorphism frequencies, or for the IGF‐2 ApaI polymorphism between female groups.Conclusion The presence of IGF‐2 ApaI polymorphism in partners of RSA women could affect IGF‐2 level of expression in placenta and embryo and represent a risk factor for RSA susceptibility.

Expression of matrix metalloproteinases during murine chorioallantoic placenta maturation.

A large body of experimental evidence supports the participation of two groups of extracellular proteases, matrix metalloproteinases (MMPs), and plasminogen activators/plasmin, in tissue remodeling in physiological and pathological invasion. In the late mouse placenta, several tissue remodeling and cell invasion processes take place. Spongiotrophoblast migration into maternal decidua, as well as decidual extracellular matrix remodeling require the coordinated action of extracellular proteolytic enzymes. Via Northern and in situ hybridization, we have analyzed the spatio-temporal expression patterns of members of the MMP family (stromelysin-3, gelatinases A and B), as well as their inhibitors TIMP-1, -2 and -3 in late murine placenta (days 10.5 to 18.5 of gestation). Gelatinase activity in placental extracts was assessed by substrate zymography. Gelatinase A and stromelysin-3 were found to be prominently expressed in decidual tissue; shortly after midpregnancy, the decidual expression patterns of gelatinase A and stromelysin-3 became overlapping with each other, as well as with the expression domain of TIMP-2. On the other hand, gelatinase B transcripts were expressed only by trophoblast giant cells at day 10.5, and were downregulated at later stages. TIMP-1 and TIMP-3 transcripts were detected in decidual periphery at day 10.5, while later the expression was restricted to the endometrial stroma and spongiotrophoblasts, respectively. The areas of stromelysin-3 expression were the same (giant trophoblasts) or adjacent (decidua) to those where urokinase (uPA) transcripts were detected, suggesting a possible cooperation between these proteinases in placental remodeling. We generated mice doubly deficient for stromelysin-3 and uPA, and report here that these mice are viable and fertile. Furthermore, these animals do not manifest obvious placental abnormalities, thereby suggesting the existence of compensatory/redundant mechanisms involving other proteolytic enzymes. Our findings document the participation of MMPs and their inhibitors in the process of late murine placenta maturation, and warrant the characterization of other members of the MMP family, like membrane type-MMPs, in this process.

BIORREGULADORES NA MORFOLOGIA E NA PRODUTIVIDADE DE FRUTOS DE TOMATEIRO CULTIVAR ÂNGELA GIGANTE

O presente trabalho teve como finalidade observar aspectos morfológicos dos frutos de tomateiro, Lycopersicon esculentum Mill. cv. Ângela Gigante, submetidos à ação de biorreguladores, com ou sem adubação adicional, e determinar as possíveis alterações na produtividade. Foram desenvolvidos, em casa de vegetação, em épocas distintas, dois ensaios. As mudas foram selecionadas e transplantadas para vasos com capacidade de 12 L de terra, contendo uma mistura de solo argiloso, areia, matéria orgânica e uma adubação mineral complementar de N, P, K. No segundo ensaio, após o transplante das mudas, além da adubação mineral complementar de N, P, K, efetuaram-se adubações adicionais (fertirrigação). Em ambos os ensaios, quando as plantas atingiram quatro folhas definitivas, realizaram-se as pulverizações com GA3 50 mg/L; NAA 100 mg/L; CCC 1.500 mg/L e SADH 3.000 mg/L. Realizaram-se, nos dois ensaios, quatro coletas de frutos maduros por planta. De modo geral, para o primeiro, o tratamento com CCC mostrou tendência em aumentar o número e a massa total de frutos por planta em relação à testemunha e aos demais tratamentos, enquanto o tratamento com NAA obteve efeito contrário. O ensaio com fertirrigação adicional não provocou alterações significativas na produção de frutos; no entanto, no tratamento com SADH, ocorreu maior incidência de anomalias. O tratamento com GA3 ocasionou, em alguns frutos, a maturação precoce da placenta em relação ao pericarpo e eventuais formações de frutos geminados. Não se observaram diferenças morfológicas significativas em relação ao comprimento e ao diâmetro médio de frutos provenientes do primeiro ensaio, porém frutos de plantas tratadas com CCC e de plantas testemunha, provindos do segundo ensaio, mostraram maior diâmetro.