Missense BICD2 variants in fetuses with congenital arthrogryposis and pterygia.

Type 2 spinal muscular atrophy with lower extremity dominance (SMALED2) is caused by bicaudal D cargo adaptor 2 (BICD2) variants. However, the SMALED2 genotype and phenotype correlation have not been thoroughly characterized. We identified de novo heterozygous BICD2 missense variants in two fetuses with severe, prenatally diagnosed multiple arthrogryposis congenita. This report provides further insights into the genetics of this rare disease.

Children's animistic beliefs toward a humanoid robot and other objects.

This study examined children's beliefs about a humanoid robot by examining their behavioral and verbal responses. We investigated whether 3- and 5-year-old children would treat the humanoid robot gently along with other objects and tools with and without a face and whether 3- and 5-year-olds would attribute moral, perceptual, and psychological properties to these targets. Although 3-year-olds did not treat objects gently or rudely, they were likely to affirm that hitting targets was acceptable despite targets having psychological and perceptual properties. Thus, 3-year-olds' perception of the targets was incongruent with their behavior toward them. Most 5-year-olds treated a robot gently and were likely to affirm the robot's psychological characteristics. Behaviors and perceptions of the robot differed between 3- and 5-year-olds. Thus, children may start believing that robots are not alive at age five, and they can distinguish them from other objects even when the latter have faces. Developmental changes in children's animistic beliefs are also discussed.

The infant-doctor relationship: an examination of infants' distress reactions in the presence of a doctor.

Fear of doctors is a common source of distress among infants; however, the underlying sources of this distress are unknown. To investigate the doctor-infant relationship, the behaviors of 61 healthy infants (176-617 days old) were observed in a simulated examination room. Their behaviors and electrocardiograms were recorded. Two groups of infants were analyzed: those who cried and those who did not. When an experimenter dressed in the doctor's attire entered the room, all 9 infants who were crying (14.8% of all infants) stopped crying, all infants gazed at the experimenter, and their mean heart rate (HR) decreased. After the auscultation started, 29.5% of all infants cried, and the HRs of infants who cried were higher than those of infants who did not cry. During the auscultation, 80.0% of infants who cried averted from the experimenter, while 34.4% of infants who did not cry. Within 5 s of gazing at the stethoscope, the number of infants who cried increased from 3 to 12, and their mean HR also increased. Our findings suggest that the fear of doctors is not due to the appearance of doctors but rather to specific actions performed by doctors, such as auscultation. Infants may regard a doctor's appearance as a source of interest. Furthermore, a stethoscope is a possible trigger for infants' crying. These behavioral observations suggest the potential for patient-centered care for infants.

Congenital anaemia associated with loss-of-function variants in DNA polymerase epsilon 1.

DNA polymerase epsilon (Pol ε), a component of the core replisome, is involved in DNA replication. Although genetic defects of Pol ε have been reported to cause immunodeficiency syndromes, its role in haematopoiesis remains unknown. Here, we identified compound heterozygous variants (p.[Asp1131fs];[Thr1891del]) in POLE, encoding Pol ε catalytic subunit A (POLE1), in siblings with a syndromic form of severe congenital transfusion-dependent anaemia. In contrast to Diamond-Blackfan anaemia, marked reticulocytopenia or marked erythroid hypoplasia was not found. Their bone marrow aspirates during infancy revealed erythroid dysplasia with strongly positive TP53 in immunostaining. Repetitive examinations demonstrated trilineage myelodysplasia within 2 years from birth. They had short stature and facial dysmorphism. HEK293 cell-based expression experiments and analyses of patient-derived induced pluripotent stem cells (iPSCs) disclosed a reduced mRNA level of Asp1131fs-POLE1 and defective nuclear translocation of Thr1891del-POLE1. Analysis of iPSCs showed compensatory mRNA upregulation of the other replisome components and increase of the TP53 protein, both suggesting dysfunction of the replisome. We created Pole-knockout medaka fish and found that heterozygous fishes were viable, but with decreased RBCs. Our observations expand the phenotypic spectrum of the Pol ε defect in humans, additionally providing unique evidence linking Pol ε to haematopoiesis.

A novel FLNA variant in a fetus with skeletal dysplasia.

Otopalatodigital spectrum disorder (OPDSD) is characterized by variable phenotypes, including skeletal dysplasia, and is caused by pathogenic variants in filamin A-encoding FLNA. FLNA variants associated with lethal OPDSD primarily alter the CH2 subdomain of the ABD of FLNA. Herein, we report a novel FLNA mutation in a fetus with severe skeletal dysplasia in a pregnant multigravida female with a history of repeated miscarriages and terminations.

Novel missense COL2A1 variant in a fetus with achondrogenesis type II.

Achondrogenesis type II (ACG2) is a lethal skeletal disorder caused by pathogenic variants in COL2A1. We present a fetus with cystic hygroma and severe shortening of the limbs at 14 weeks of gestation. We performed postnatal genetic analysis of the parents and fetus to diagnose the disease. A novel missense variant of COL2A1 [NM_001844.5: c.2987G>A, (p. Gly996Asp)] was identified, which led to the ACG2 diagnosis.

Glucocorticoids increase the risk of preterm premature rupture of membranes possibly by inducing ITGA8 gene expression in the amnion.

INTRODUCTION: Maternal glucocorticoid exposure increases the risk of preterm delivery; however, the association between glucocorticoids and preterm premature rupture of membranes (pPROM)-a direct cause of preterm delivery-has rarely been investigated. METHODS: To examine this association, we evaluated the clinical data of patients with systemic lupus erythematosus (SLE). Mechanism analysis was performed in both human amnion-derived mesenchymal cells (as a model for fetal membranes) and the amnion from SLE patients. We characterized the effects of glucocorticoids on the amnion in both models through comprehensive gene expression profiling and by electric cell-substrate impedance sensing in the mesenchymal cells. RESULTS: The average glucocorticoid dose in cases with pPROM (13.3 mg/day, n = 10) was significantly higher than in those without pPROM (8.5 mg/day, n = 65; P < 0.01) among pregnant patients with well-controlled SLE (SLEDAI <4, n = 75); however, we did not observe a statistically significant difference in it between cases with or without chorioamnionitis. Glucocorticoid-treated human amnion mesenchymal cells showed decreased electric resistance between cells, indicating increased permeability. Differentially expressed genes upon glucocorticoid treatment were significantly enriched with cell adhesion-related genes. Among them, ITGA8 was strikingly induced in both the amnion mesenchymal cells and in amnion derived from patients with SLE. DISCUSSION: We observed an association between glucocorticoids and pPROM with non-infectious etiology. Our findings indicate that glucocorticoids increase amnion permeability and modulate cell-adhesion related genes. ITGA8 represents a primary molecule that triggers pPROM through fibrotic remodeling and preventing resealing of the rupture site in fetal amnion.

Pregnancy Outcome and Postnatal Chromosome Analysis of the Cord Blood and Chorionic Villi in Two Cases after Intrauterine Transfer of Mosaic Aneuploid Blastocysts.

The long-term prognosis and genetic mechanism of pregnancy after intrauterine mosaic aneuploid blastocyst transfer remain unknown. We report the case of two babies after the aforementioned procedure and chromosomal analysis of their cord blood and chorionic villi. Case Report 1. A 41-year-old primipara, with two previous spontaneous abortions, was pregnant after intrauterine transfer of a blastocyst carrying 40% mosaicism of long-arm monosomy of chromosome 5. The amniocentesis results were 46,XX. A cesarean section was performed at 39 weeks. The female infant was 3,315 g at birth. Case Report 2. A 44-year-old primipara, with two spontaneous abortions, was pregnant after intrauterine transfer of a blastocyst carrying 40% mosaicism of long-arm monosomy of chromosome 9 and monosomy of chromosome 14. After genetic counselling, she decided not to undergo amniocentesis. No abnormalities were found by ultrasound. A cesarean section was performed at 38 weeks. The male infant was 3,340 g at birth. Chromosome analyses of postnatal cord blood and chorionic villi were performed using SNP arrays. The cord blood and chorionic villi showed no chromosomal structural abnormalities or mosaicism. For both, no disorders were observed at 10 months of age. We experienced the birth of babies after intrauterine transfer of mosaic aneuploid blastocysts.

Collection of 2429 constrained headshots of 277 volunteers for deep learning.

Deep learning has rapidly been filtrating many aspects of human lives. In particular, image recognition by convolutional neural networks has inspired numerous studies in this area. Hardware and software technologies as well as large quantities of data have contributed to the drastic development of the field. However, the application of deep learning is often hindered by the need for big data and the laborious manual annotation thereof. To experience deep learning using the data compiled by us, we collected 2429 constrained headshot images of 277 volunteers. The collection of face photographs is challenging in terms of protecting personal information; we therefore established an online procedure in which both the informed consent and image data could be obtained. We did not collect personal information, but issued agreement numbers to deal with withdrawal requests. Gender and smile labels were manually and subjectively annotated only from the appearances, and final labels were determined by majority among our team members. Rotated, trimmed, resolution-reduced, decolorized, and matrix-formed data were allowed to be publicly released. Moreover, simplified feature vectors for data sciences were released. We performed gender and smile recognition by building convolutional neural networks based on the Inception V3 model with pre-trained ImageNet data to demonstrate the usefulness of our dataset.

Improved library preparation protocols for amplicon sequencing-based noninvasive fetal genotyping for RHD-positive D antigen-negative alleles.

OBJECTIVE: We aimed to simplify our fetal RHD genotyping protocol by changing the method to attach Illumina's sequencing adaptors to PCR products from the ligation-based method to a PCR-based method, and to improve its reliability and robustness by introducing unique molecular indexes, which allow us to count the numbers of DNA fragments used as PCR templates and to minimize the effects of PCR and sequencing errors. RESULTS: Both of the newly established protocols reduced time and cost compared with our conventional protocol. Removal of PCR duplicates using UMIs reduced the frequencies of erroneously mapped sequences reads likely generated by PCR and sequencing errors. The modified protocols will help us facilitate implementing fetal RHD genotyping for East Asian populations into clinical practice.

Isolation and characterization of fetal nucleated red blood cells from maternal blood as a target for single cell sequencing-based non-invasive genetic testing.

PURPOSE: Although non-invasive prenatal testing (NIPT) based on cell-free DNA (cfDNA) in maternal plasma has been prevailing worldwide, low levels of fetal DNA fraction may lead to false-negative results. Since fetal cells in maternal blood provide a pure source of fetal genomic DNA, we aimed to establish a workflow to isolate and sequence fetal nucleated red blood cells (fNRBCs) individually as a target for NIPT. METHODS: Using male-bearing pregnancy cases, we isolated fNRBCs individually from maternal blood by FACS, and obtained their genomic sequence data through PCR screening with a Y-chromosome marker and whole-genome amplification (WGA)-based whole-genome sequencing. RESULTS: The PCR and WGA efficiencies of fNRBC candidates were consistently lower than those of control cells. Sequencing data analyses revealed that although the majority of the fNRBC candidates were confirmed to be of fetal origin, many of the WGA-based genomic libraries from fNRBCs were considered to have been amplified from a portion of genomic DNA. CONCLUSIONS: We established a workflow to isolate and sequence fNRBCs individually. However, our results demonstrated that, to make cell-based NIPT targeting fNRBCs feasible, cell isolation procedures need to be further refined such that the nuclei of fNRBCs are kept intact.

Novel TNFAIP3 microdeletion in a girl with infantile-onset inflammatory bowel disease complicated by a severe perianal lesion.

A20 haploinsufficiency (HA20), a disease caused by loss-of-function TNFAIP3 mutations, manifests various autoinflammatory and/or autoimmune symptoms. Some cases of HA20 were initially diagnosed as very early onset inflammatory bowel disease (VEO-IBD). We performed whole-exome sequencing (WES) for a Japanese girl with infantile-onset IBD and a severe perianal lesion and detected a novel de novo 119 kb microdeletion containing only TNFAIP3 (arr[GRCh37] 6q23.3(138125829_138244816) × 1).

Effects of high-intensity interval exercise under hyperoxia on HSP27 and oxidative stress responses.

Hypoxia in working muscles during exercise may be associated with increased oxidative stress. Inhalation of hyperoxic gas diminishes the hypoxia within working muscles during exercise. Exposure to hyperoxia increases the expression of the antioxidant HSP27. We investigated the effects of acute high-intensity interval exercise (HIE) under hyperoxia on HSP27 levels and oxidative stress responses. Eight male subjects participated in two experiments: 1) normoxic HIE (NHIE) and 2) hyperoxic (60 % oxygen) HIE (HHIE). HIE consisted of four 30-s all-out cycling bouts with 4-min rest between bouts. Levels of serum oxidative stress markers (d-ROMs and LPO), HSP27, BAP, IL-6, and TNF-α significantly increased after both trials. The HIE-induced changes in d-ROMs, LPO, and HSP27 levels were significantly lower in the HHIE trial than in the NHIE trial. These findings suggest that inhaling hyperoxic gas during exercise might diminish oxidative stress induced by all-out HIE.

The Categorization of Objects With Uniform Texture at Superordinate and Living/Non-living Levels in Infants: An Exploratory Study.

Human infants can categorize objects at various category levels (e.g., as a dog, animal, or living thing). It is crucial to understand how infants learn about the relationships between objects. This study investigated whether 4- to 11-month-old infants can categorize modeled objects at superordinate and living/non-living levels. In this experiment, we presented modeled objects with a uniform texture constructed by a 3D printer in animal, vegetable/fruit, vehicle, and tool categories and measured the time taken to examine novel categories. We investigated infants' categorization abilities using familiarization/novelty-preference tasks and their pre-linguistic development based on information from their parents. The analyses examined whether infants dedicated more examination time to objects in the new category at superordinate and living/non-living levels for each month of age. The results revealed that the examination time among 4- and 5-month-olds was at chance levels for both superordinate and living/non-living levels, while at 7 months, they showed high preference for the novel category at both category levels. For the superordinate level, the strength of response to living objects increased with linguistic development, while the strength of response to non-living objects did not depend on linguistic development. This indicates that the superordinate-level categorization of living objects depends on both perceptual information and linguistic ability. For the living/non-living level, the examination time for non-living objects increased with linguistic development. This implies that the recognition of non-living objects may depend on the development of object knowledge. The current study suggests that infants can recognize categories at an abstract level before the acquisition of linguistic representations while the category levels that infants can categorize objects are different for living/non-living objects. This may imply that infants learn the concepts of living/non-living via different mechanisms.

Monochorionic diamniotic twins of discordant external genitalia with 45,X/46,XY mosaicism.

BACKGROUND: Monozygotic twins with 45,X/46,XY mosaicism, discordant for phenotypic sex, are extremely rare. METHODS: This report describes the clinical findings of a rare case of 45,X/46,XY mosaicism in monozygotic twins with different external genitalia. Single nucleotide polymorphism (SNP) array analysis, performed by collecting DNA from each umbilical cord, showed identical SNPs in the autosomal chromosomes of both fetuses. RESULTS: Chorionic villus sampling of a 37-year-old primigravida carrying monozygotic twins revealed a 45,X/46,XY karyotype. Autopsy of the aborted fetuses revealed a penis and testes on one fetus and a vagina, uterus, and ovaries in the other fetus--which also had severe cystic hygroma. Cell counting using fluorescence in situ hybridization with XY probes (XY-FISH) showed 20% and 80% abundance of 45,X cells in the internal genitalia, liver, heart, lung, adrenal gland, bone marrow, and spine of the male and female fetuses, respectively. CONCLUSION: These results indicated that the fetuses were genetically monozygotic twins and their different degrees of mosaicism may have resulted in different genital phenotypes.

Object Categorization Processing Differs According to Category Level: Comparing Visual Information Between the Basic and Superordinate Levels.

Object category levels comprise a crucial concept in the field of object recognition. Specifically, categorization performance differs according to the category level of the target object. This study involved experiments with two types of stimulus sequences (i.e., forward condition: presenting the target name before the line-drawing stimulus; and reverse condition: presenting the target name after the line-drawing stimulus) for both basic- and superordinate-level categorizations. Adult participants were assigned to each level and asked to judge whether briefly presented stimuli included the same object and target name. Here, we investigated how the category level altered the categorization process. We conducted path analyses using a multivariate multiple regression model, and set our variables to investigate whether the predictors affected the categorization process between two types of stimulus sequence. Dependent variables included the measures of performance (i.e., reaction time, accuracy) for each categorization task. The predictors included dimensions and shapes of the line-drawings, such as primary and local shape information, shape complexity, subject estimation, and other shape variables related to object recognition. Results showed that the categorization process differed according to shape properties between conditions only for basic-level categorizations. For the forward condition, the bottom-up processing of primary visual information depended on matches with stored representations for the basic-level category. For the reverse condition at the basic-level category, decisions depended on subjective ratings in terms of object-representation accessibility. Finally, superordinate-level decisions depended on higher levels of visual information in terms of complexity, regardless of the condition. Thus, the given category level altered the processing of visual information for object recognition in relation to shape properties. This indicates that decision processing for object recognition is flexible depending on the criteria of the processed objects (e.g., category levels).

Epitranscriptomic profiling in human placenta: N6-methyladenosine modification at the 5'-untranslated region is related to fetal growth and preeclampsia.

Intracellular mRNA levels are not always proportional to their respective protein levels, especially in the placenta. This discrepancy may be attributed to various factors including post-transcriptional regulation, such as mRNA methylation (N6-methyladenosine: m6A). Here, we conducted a comprehensive m6A analysis of human placental tissue from neonates with various birth weights to clarify the involvement of m6A in placental biology. The augmented m6A levels at the 5'-untranslated region (UTR) in mRNAs of small-for-date placenta samples were dominant compared to reduction of m6A levels, whereas a decrease in m6A in the vicinity of stop codons was common in heavy-for-date placenta samples. Notably, most of these genes showed similar expression levels between the different birth weight categories. In particular, preeclampsia placenta samples showed consistently upregulated SMPD1 protein levels and increased m6A at 5'-UTR but did not show increased mRNA levels. Mutagenesis of adenosines at 5'-UTR of SMPD1 mRNAs actually decreased protein levels in luciferase assay. Collectively, our findings suggest that m6A both at the 5'-UTR and in the vicinity of stop codon in placental mRNA may play important roles in fetal growth and disease.

The outcomes and risk factors of fetal bradycardia associated with external cephalic version.

OBJECTIVE: The objective of this study is to assess the outcomes and risk factors of fetal bradycardia after external cephalic version (ECV). METHODS: We performed a retrospective study of women who underwent ECV after 35 weeks of gestation in 2010-2016. We assessed the birth outcomes, including umbilical cord artery pH, according to the duration of fetal bradycardia and the risk factors for bradycardia. RESULTS: Among 390 cases, 189 (48.5%) cases showed fetal bradycardia during or immediately after ECV. The duration of fetal bradycardia was <1 min (n = 82, 43.4%), <5 min (n = 168, 88.9%); and <10 min (n = 186, 98.4%). All cases showed a good prognosis. Fetal bradycardia lasting >10 min occurred in three cases; emergency cesarean section was performed in each case, with delivery after 12-4 min of bradycardia. Two of three cases showed low Apgar scores at 5 min, with an umbilical cord arterial pH of <7.1. Lower maternal BMI and a prolonged ECV procedure were significantly associated with bradycardia (p for trend: .016 and .015, respectively). CONCLUSIONS: Fetal bradycardia lasting >10 min after ECV was a risk factor for asphyxia. Thus, delivery should be completed within 10 min after bradycardia. A low maternal BMI and a prolonged ECV procedure were risk factors for bradycardia after ECV.

Task Difficulty Makes 'No' Response Different From 'Yes' Response in Detection of Fragmented Object Contours.

Two-alternative forced choice tasks are often used in object detection, which regards detecting an object as a 'yes' response and detecting no object as a 'no' response. Previous studies have suggested that the processing of yes/no responses arises from identical or similar processing. In this study, we investigated the difference of processing between detecting an object ('yes' response) and not detecting any object ('no' response) by controlling the task difficulty in terms of fragment length and stimulus duration. The results indicated that a 'yes' response depends on accurate and stable decisions through grouping processing, and a 'no' response might involve two distinct processing, including accurate decisions and intuitive decisions. Accurate decisions of 'no' may arise after the rejection of a 'yes' response with grouping processing, which is an accurate but slow response in an easy task. Intuitive decisions of 'no' arise as the result of breaking down the decision process when the received information was insufficient for grouping processing in a difficult task. Therefore, intuitive decisions of 'no' arise quickly but are inaccurate. The different processes associated with yes/no responses were discussed in terms of the hierarchal structure of object recognition, especially with respect to receiving information and grouping.

Placental Development and Nutritional Environment.

The placenta is considered to have developed recently in mammalian evolution. While the fundamental function of the placenta, i.e., providing nutrients and oxygen to the fetus and receiving waste products, is the same in all mammals, the morphology of the placenta varies substantially in a species-dependent manner. Therefore, considerable interest exists in understanding placental development and function in mammals from a molecular biological viewpoint. Numerous recent studies have shown that various environmental factors before and during pregnancy, including nutrition, affect placental formation and function and that alterations in placental formation and function can influence the developing fetus and the offspring after birth. To date, the relationship between nutrition and the placenta has been investigated in several species, various model organisms, and humans. In this chapter, we discuss the current knowledge of the placenta and the epigenome and then highlight the effects of nutrition during pregnancy on the placenta and the fetus and on the offspring after birth.