Stillborn Fetuses Research Articles

Vascular Placental Pathology and Cardiac Structure in Stillborn Fetuses.

Adverse pregnancy outcomes, including preterm birth and preeclampsia, are associated with worse cardiovascular outcomes for offspring. Examination of the placenta is important for understanding how the prenatal period shapes long-term cardiovascular health. We sought to investigate the association between placental vascular malperfusion and fetal cardiac structure. Data obtained from the Stillbirth Collaborative Research Network included stillbirths with placental pathology and autopsy. Stillbirths were classified in two ways: based on severity of placental maternal vascular malperfusion (MVM) and based on cause of death (MVM, fetal vascular malperfusion (FVM), or acute infection/controls). Organ weight and heart measures were standardized by gestational age (GA) and compared across groups. We included 329 stillbirths in the analysis by MVM severity and 76 in the analysis by cause of death (COD). While z-scores for most organ weights/heart measures were smaller when COD was attributed to MVM as compared with FVM or controls, heart weight and brain weight z-score did not differ by COD (p>0.05). In analyses accounting for body size, the difference between heart and body weight z-score was -0.05 (SD: 0.53) among those with MVM as a COD and -0.20 (SD: 0.95) among those with severe MVM. Right and left ventricle thicknesses and tricuspid, pulmonary, mitral, and aortic valve circumferences were consistently as expected or larger than expected for GA and body weight. In analysis investigating severity of MVM, those with the most severe MVM had heart measures that were as expected or larger than expected for body weight while those with only mild to moderate MVM had heart measures that were generally small relative to body weight. When assessed as COD or based on severity, MVM was associated with heart measures that were as expected or larger than expected for GA and body weight, indicating that possible heart sparing.

Oropouche Virus (OROV) in Pregnancy: An Emerging Cause of Placental and Fetal Infection Associated with Stillbirth and Microcephaly following Vertical Transmission.

Oropouche virus (OROV) is an emerging arbovirus endemic in Latin America and the Caribbean that causes Oropouche fever, a febrile illness that clinically resembles some other arboviral infections. It is currently spreading through Brazil and surrounding countries, where, from 1 January to 1 August 2024, more than 8000 cases have been identified in Bolivia, Brazil, Columbia, and Peru and for the first time in Cuba. Travelers with Oropouche fever have been identified in the United States and Europe. A significant occurrence during this epidemic has been the report of pregnant women infected with OROV who have had miscarriages and stillborn fetuses with placental, umbilical blood and fetal somatic organ samples that were RT-PCR positive for OROV and negative for other arboviruses. In addition, there have been four cases of newborn infants having microcephaly, in which the cerebrospinal fluid tested positive for IgM antibodies to OROV and negative for other arboviruses. This communication examines the biology, epidemiology, and clinical features of OROV, summarizes the 2023-2024 Oropouche virus epidemic, and describes the reported cases of vertical transmission and congenital infection, fetal death, and microcephaly in pregnant women with Oropouche fever, addresses experimental animal infections and potential placental pathology findings of OROV, and reviews other bunyavirus agents that can cause vertical transmission. Recommendations are made for pregnant women travelling to the regions affected by the epidemic.

Rising Stillbirth Rates Related to Congenital Syphilis in the United States From 2016 to 2022.

To identify trends in stillbirth rates attributed to congenital syphilis in the United States by describing congenital syphilis-related stillbirths and comparing characteristics of pregnant people who had congenital syphilis-related stillbirths with those of people who had preterm and full-term liveborn neonates with congenital syphilis. Cases of congenital syphilis reported to the Centers for Disease Control and Prevention during 2016-2022 were analyzed and categorized as stillbirth, preterm live birth (before 37 weeks of gestation), or term live birth (37 weeks or later). Cases with unknown vital status or gestational age were excluded. Frequencies were calculated by pregnancy outcome, including pregnant person demographics; receipt of prenatal care; syphilis stage and titer; and timing of prenatal care, testing, and treatment. Overall, 13,393 congenital syphilis cases with vital status and gestational age were reported; of these, 853 (6.4%) were stillbirths. The number of congenital syphilis-related stillbirths increased each year (from 44 to 231); the proportion of congenital syphilis cases resulting in stillbirth ranged from 5.2% to 7.5%. Median gestational age at delivery for stillborn fetuses was 30 weeks (interquartile range 26-33 weeks). People with congenital syphilis-related stillbirths were more likely to have titers at or above 1:32 (78.9% vs 45.5%; P<.001) and to have received no prenatal care (58.4% vs 33.1%; P<.001) than people with liveborn neonates with congenital syphilis. The risk of stillbirth was twice as high in persons with secondary syphilis compared with those with primary syphilis (11.5% vs 5.7%, risk ratio 2.00; 95% CI, 1.27-3.13). Across all congenital syphilis cases, 34.2% of people did not have a syphilis test at their first prenatal visit. Stillbirths occurred in more than 1 in 20 pregnancies complicated by congenital syphilis. Risk factors for stillbirth included high titers, secondary stage, and lack of prenatal care. If the prevalence of congenital syphilis continues to rise, there will be a corresponding increase in the overall stillbirth rate nationally.

In Situ Analyses of Placental Inflammatory Response to SARS-CoV-2 Infection in Cases of Mother-Fetus Vertical Transmission.

It has been shown that vertical transmission of the SARS-CoV-2 strain is relatively rare, and there is still limited information on the specific impact of maternal SARS-CoV-2 infection on vertical transmission. The current study focuses on a transcriptomics analysis aimed at examining differences in gene expression between placentas from mother-newborn pairs affected by COVID-19 and those from unaffected controls. Additionally, it investigates the in situ expression of molecules involved in placental inflammation. The Papa Giovanni XXIII Hospital in Bergamo, Italy, has recorded three instances of intrauterine transmission of SARS-CoV-2. The first two cases occurred early in the pandemic and involved pregnant women in their third trimester who were diagnosed with SARS-CoV-2. The third case involved an asymptomatic woman in her second trimester with a twin pregnancy, who unfortunately delivered two stillborn fetuses due to the premature rupture of membranes. Transcriptomic analysis revealed significant differences in gene expression between the placentae of COVID-19-affected mother/newborn pairs and two matched controls. The infected and control placentae were matched for gestational age. According to the Benjamani-Hochberg method, 305 genes met the criterion of an adjusted p-value of less than 0.05, and 219 genes met the criterion of less than 0.01. Up-regulated genes involved in cell signaling (e.g., CCL20, C3, MARCO) and immune response (e.g., LILRA3, CXCL10, CD48, CD86, IL1RN, IL-18R1) suggest their potential role in the inflammatory response to SARS-CoV-2. RNAscope® technology, coupled with image analysis, was utilized to quantify the surface area covered by SARS-CoV-2, ACE2, IL-1β, IL-6, IL-8, IL-10, and TNF-α on both the maternal and fetal sides of the placenta. A non-statistically significant gradient for SARS-CoV-2 was observed, with a higher surface coverage on the fetal side (2.42 ± 3.71%) compared to the maternal side (0.74 ± 1.19%) of the placenta. Although not statistically significant, the surface area covered by ACE2 mRNA was higher on the maternal side (0.02 ± 0.04%) compared to the fetal side (0.01 ± 0.01%) of the placenta. IL-6 and IL-8 were more prevalent on the fetal side (0.03 ± 0.04% and 0.06 ± 0.08%, respectively) compared to the maternal side (0.02 ± 0.01% and 0.02 ± 0.02%, respectively). The mean surface areas of IL-1β and IL-10 were found to be equal on both the fetal (0.04 ± 0.04% and 0.01 ± 0.01%, respectively) and maternal sides of the placenta (0.04 ± 0.05% and 0.01 ± 0.01%, respectively). The mean surface area of TNF-α was found to be equal on both the fetal and maternal sides of the placenta (0.02 ± 0.02% and 0.02 ± 0.02%, respectively). On the maternal side, ACE-2 and all examined interleukins, but not TNF-α, exhibited an inverse mRNA amount compared to SARS-CoV-2. On the fetal side, ACE-2, IL-6 and IL-8 were inversely correlated with SARS-CoV-2 (r = -0.3, r = -0.1 and r = -0.4, respectively), while IL-1β and IL-10 showed positive correlations (r = 0.9, p = 0.005 and r = 0.5, respectively). TNF-α exhibited a positive correlation with SARS-CoV-2 on both maternal (r = 0.4) and fetal sides (r = 0.9) of the placenta. Further research is needed to evaluate the correlation between cell signaling and immune response genes in the placenta and the vertical transmission of SARS-CoV-2. Nonetheless, the current study extends our comprehension of the molecular and immunological factors involved in SARS-CoV-2 placental infection underlying maternal-fetal transmission.

High Rates of Miscarriage and Stillbirth among Pregnant Women with Clade I Mpox (Monkeypox) Are Confirmed during 2023-2024 DR Congo Outbreak in South Kivu Province.

Mpox (monkeypox) is a neglected tropical disease that has received increased attention since the multi-nation outbreak that began in 2022. The virus is endemic in West and Central Africa, where the Democratic Republic of the Congo (DRC) is the most affected country. Clade I monkeypox virus (MPXV) infection is endemic in the DRC and has an overall case fatality rate of 10.6% among children and adults. A study conducted in Sankuru Province, DRC, from 2007 to 2011 demonstrated that 75% of pregnant women with mpox had miscarriages or stillbirth. Further analysis of a stillborn fetus showed that MPXV could infect both the placenta and fetus, causing congenital infection. No additional cases of Clade I MPXV in pregnant women were reported until a new outbreak occurred in South Kivu Province during 2023 and 2024. Eight pregnant women having Clade I MPXV infection were identified, of whom four had either miscarriages or stillbirth, representing a 50% fetal mortality rate. These reports confirm previous data from the DRC that indicate the capability of Clade I MPXV to affect the fetus, causing congenital infection and fetal loss in a high percentage of cases. In this article, we review both past and new data from the DRC on the effects of Clade I MPXV during pregnancy and discuss the association of mpox with fetal loss.

Genetic Characterization of Palyam Serogroup Viruses Isolated in Japan from 1984 to 2018 and Development of a Real-Time RT-PCR Assay for Broad Detection of Palyam Serogroup Viruses and Specific Detection of Chuzan (Kasba) and D'Aguilar Viruses.

We performed whole genome sequencing (WGS) of 15 Palyam serogroup virus (PALV) strains isolated from cattle or Culicoides biting midges in Japan from 1984 to 2018. We found that the PALV strains consisted of Chuzan (Kasba) virus (CHUV), D'Aguilar virus (DAGV), Bunyip Creek virus, and another PALV, Marrakai virus (MARV). The Japanese MARV strains isolated in 1997 were closely related to Australian PALV strains isolated in 1968-1976 in genome segments 2 and 10, but they were most closely related to other Japanese PALV strains in the other genome segments. Our data suggest that the Japanese MARV strains were reassortant viruses between Asian and Australian PALVs. In addition to the WGS, we developed a real-time reverse-transcription polymerase chain reaction assay that can broadly detect PALV and specifically detect CHUV and DAGV, utilizing the data obtained by the WGS in this study. We detected the DAGV gene in bovine stillborn fetuses and congenitally abnormal calves in 2019 using the newly developed assay. To our knowledge, this is the first report of isolation of MARV outside of Australia and the first report of detection of PALV in bovine fetuses or calves with congenital abnormality outside of Africa.

Identifying Risk of Stillbirth Using Machine Learning

(Am J Obstet Gynecol. 2023;229:327:e1–16) A tool that could help identify women at risk for stillbirth would be a welcome addition to clinical practice. Machine learning models, which can look at relationships between variables in large data sets, was investigated in this article. This study’s aim was to refine a machine learning model to predict stillbirth using data available prior to the gestational age of viability (22 to 24 wk). The researchers performed a secondary analysis of data in the Stillbirth Collaborative Research Network (SCRN), a study database of 982 stillbirths and 3000 representative live births from 2006 to 2009. The study included patients who delivered a live or stillborn fetus at &gt;18 weeks’ gestation from 59 hospitals in 5 states (Georgia, Massachusetts, Rhode Island, Texas, and Utah). The data set included 6000 measures collected from interviews with the mother, postmortem pathological exam, cause of death analysis, and medical record Abstractsion. Subsequently 101 risk factor variables were identified and used in the model. These included maternal and biological father demographics, family history, maternal health history, prenatal lab data, fetal genetics, second-trimester screen, prenatal care history, ultrasound, and social determinants of health.

16 Effects of fiber supplementation during gestation on sow and piglet performance

Abstract Our objective was to determine the effects of fiber supplementation during gestation on sow and piglet performance. A total of 200 PIC Camborough P1+ sows were used. At breeding, sows were balanced by body weight (BW) within parity and randomly allotted to 1 of 2 dietary treatments, which consisted of 1) Control; corn-soybean meal gestation diet; and 2) Fiber; inclusion of soybean hulls in the formulation at a level to yield a dietary crude-fiber content 6%. Females were housed in individual stalls throughout the entire gestation. At d 90 of gestation, serum glucose, fecal dry matter, fecal volatile fat acids (acetate, propionate, butyrate, isovalerate, and valerate), urinary traits (color, turbidity, density, pH, and presence of nitrate or blood), and frequency of stereotypic behavior (sham chewing, teeth grinding, bar biting, and floor licking) was accessed. Perineal scoring was visually evaluated at d 105 and 110 of gestation. Farrowing traits (farrowing length, and number of piglets born alive, stillborn, and mummified fetuses) and piglet vitality (meconium presence, umbilical cord integrity, oxygenation, and heart rate) were individually monitored. Individual piglet BW was obtained within 12 h of birth and after 24 h for colostrum intake, and at weaning. Sow daily lactation feed intake was registered. Data were analyzed using generalized linear mixed models with the Glimmix procedure of SAS with sow serving as the experimental unit. There was no evidence (P &amp;gt; 0.10) for differences in sow BW at d1, d112 of gestation, and at weaning. Sows fed the greater fiber diet showed lower levels of post-prandial glucose (-6%; P &amp;lt; 0.10), darker urine (+33%; P &amp;lt; 0.10) with greater pH (+5%; P &amp;lt; 0.05), and greater levels of fecal butyrate (+62%; P &amp;lt; 0.05) and isovalerate (+15%; P &amp;lt; 0.10) at the end of gestation (d 90). Feces from sows fed the control diet were drier (P &amp;lt; 0.05) at d 90 of gestation. Fiber supplementation reduced the frequency of teeth grinding (-52%; P &amp;lt; 0.10), bar biting (-74%; P &amp;lt; 0.05), and floor licking (-65%; P 0.10) behaviors. There was no evidence for treatment differences (P &amp;gt; 0.10) for perineal scores. Piglet vitality was not affected by the treatments, except for a marginal increase in blood oxygen saturation for piglets from sows fed the greater fiber diet (+1%; P &amp;lt; 0.10). Treatment did not impact (P &amp;gt; 0.10) farrowing duration, number of total piglets born, born alive, stillborn, and mummified, nor average piglet birth weight and colostrum intake. Sows fed diet with fiber supplementation during gestation had a marginal increase in average daily feed intake during lactation (+0.50kg/d; P = 0.08), which resulted in improved piglet BW gain (P &amp;lt; 0.05) and weaning weights (+0.20kg; P &amp;lt; 0.05). In summary, providing increased level of fiber supplementation via soybean hulls to gestating diets did not impact sow reproductive performance. Beneficial effects were observed for welfare lactation feed intake, piglet BW gain and weaning weights.

Porcine circovirus 2d associated with stillborn mummified foetus of pig

Recurrent occurrence of stillborn piglets was noticed along with live births during farrowing in pigs in an organized pig farm of Uttar Pradesh. On detailed post mortem examination, externally, the foetus was found to be partially dehydrated and dark brown in colour with shrunken eyes. Internally, visceral organs including lungs, heart, kidneys, liver, spleen and intestine were partially dehydrated and dark brown in colour. Upon molecular investigation, the pooled tissue samples were found positive for porcine circovirus 2 (PCV2) infection using PCR technique and further confirmed the PCV2d genotype by DNA sequencing. Other most possible infections including porcine parvoviral infection (PPV), PCV3 and porcine respiratory and reproductive syndrome virus infection (PRRS) were found negative using PCR and RT-PCR techniques, respectively. This result is suggestive of role of PCV2d as a possible etiological agent to cause stillbirth and mummification of porcine foetuses.

Estudo comparativo de análise ultrassonográfica e anatomopatológica de fetos caninos apresentando hidropisia fetal

Fetal hydrops is a pathological condition that is incompatible with the life of the affected animal and can lead to fetal distortions. The diagnosis is frequent in routine dog neonatology and can be made by gestational ultrasound and/or necroscopic study. To carry out this study, we analyzed three stillborn fetuses from French bulldog dogs with hydrops using echographic and anatomopathological techniques. Ultrasound was used to check for the presence of fluid in tissues such as the subcutaneous tissue, the abdominal and thoracic cavity and the intraencephalic tissue. The necroscopic study revealed morphological changes in the brain and the presence of hydropericardium. This confirmed that ultrasound can accurately identify subcutaneous and intracavitary fluids, as well as verify the brain morphology of fetuses. However, there is evidence of limitations to the ultrasound technique, given that alterations such as hydropericardium and encephalic morphological changes were not detected during the ultrasound study.

Preimplantation genetic testing as a means of preventing hereditary congenital myasthenic syndrome caused by RAPSN.

Congenital myasthenic syndrome is a heterogeneous group of inherited neuromuscular transmission disorders. Variants in RAPSN are a common cause of CMS, accounting for approximately 14%-27% of all CMS cases. Whether preimplantation genetic testing for monogenic disease (PGT-M) could be used to prevent the potential birth of CMS-affected children is unclear. Application of WES (whole-exome sequencing) for carrier testing and guidance for the PGT-M in the absence of a genetically characterized index patient as well as assisted reproductive technology were employed to prevent the occurrence of birth defects in subsequent pregnancy. The clinical phenotypes of stillborn fetuses were also assessed. The family carried two likely pathogenic variants in RAPSN(NM_005055.5): c.133G>A (p.V45M) and c.280G>A (p.E94K). And the potential birth of CMS-affected child was successfully prevented, allowing the family to have offspring devoid of disease-associated variants and exhibiting a normal phenotype. This report constitutes the first documented case of achieving a CMS-free offspring through PGT-M in a CMS-affected family. By broadening the known variant spectrum of RAPSN in the Chinese population, our findings underscore the feasibility and effectiveness of PGT-M for preventing CMS, offering valuable insights for similarly affected families.

A probable cluster of premature birth and stillbirth caused by Leptospira interrogans serogroup Hebdomadis in an integrated swine farm in Nagasaki Prefecture, Japan.

In an integrated swine farm with 135 sows in Nagasaki Prefecture, premature births and stillbirths were observed in six sows in June and July 2023, and their etiology was investigated. Leptospiral flaB gene was detected in the kidney of one stillborn fetus and the placenta of its sow by nested PCR. Multilocus sequence typing revealed that the infecting strain was Leptospira interrogans ST118. In addition, leptospiral antigens were detected in lesions of the above tissues by immunohistochemical staining. The examined sow and several other sows in the farm had antibodies against serogroup Hebdomadis. These results suggested that L. interrogans serogroup Hebdomadis ST118 was the causative agent of premature births and stillbirths that occurred in this farm.

Filial cannibalism in rainbow boas (Epicrates cenchria and E. assisi)

Cannibalism, or intraspecific predation, refers to the practice in which an individual completely or partially consumes another conspecific. Filial cannibalism occurs when parents consume their own offspring, whether they are alive or stillborn. This study describes filial cannibalism behavior in just one female of each species, Amazon rainbow boa (Epicrates cenchria) and the Caatinga rainbow boa (E. assisi). We observed that female E. cenchria and E. assisi only consumed atretic eggs. A female E. cenchria consumed two out of the eight atretic eggs she delivered, while sparing the four neonates and the only stillborn fetus in her litter. A female E. assisi consumed two out of the four atretic eggs and attempted to ingest the amniotic sacs and liquid yolk throughout the entire observation, sparing two neonates. This is the first documented record of such behavior in E. assisi.

Mirror syndrome associated with fetal cardiomyopathy

Mirror syndrome is a rare condition of generalized maternal oedema caused by fetal hydrops. A 37-year-old patient was admitted to our hospital because of suspected mirror syndrome caused by fetal cardiomyopathy. At 26th week of gestation patient developed bilateral pulmonary oedema as her condition rapidly deteriorated. Consequently, preterm labor was induced, percutaneous evacuation of fetal ascites was performed, and the patient finally vaginally delivered stillborn fetus. Although the initial postpartum period was severely complicated by hemorrhage, the condition of the patient significantly improved later, and she was discharged seven days after delivery. We believe this case is worth presenting due to its rarity and significant perinatal and obstetric challenges in treatment of those patients. Furthermore, preimplantation genetic testing could be performed to prevent at least some of the cases.

Congenital Mpox Syndrome (Clade I) in Stillborn Fetus after Placental Infection and Intrauterine Transmission, Democratic Republic of the Congo, 2008.

We report the autopsy pathology findings of a 21-week stillborn fetus with congenital mpox syndrome that occurred in the Democratic Republic of the Congo in 2008. The fetus acquired mpox from the mother after intrauterine transplacental monkeypox virus transmission. We confirmed monkeypox virus infection in the mother, fetus, and placenta by using a monkeypox virus-specific quantitative PCR. Subtyping of the virus was not performed, but the mother and fetus were almost certainly infected with the clade I variant that was endemic in the Democratic Republic of the Congo at the time. Risk for intrauterine infection appears to differ between virus clades, but clinicians should be aware of potential for intrauterine monkeypox virus transmission among pregnant persons during ongoing and future mpox outbreaks.

Potential risk factors for fetal loss due to umbilical cord torsion in the mare

Equine pregnancy failure can occur throughout gestation with etiologies varying by region. In Europe, umbilical cord torsion (UCT) has been reported to account for up to 52% of abortions and stillbirths. Little is known about what causes excessive twisting of the umbilical cord (UC), and up till now, only the length of the UC has been identified as a risk factor for UCT. The aim of the present study was to determine the association of potential maternal and fetal risk factors with UCT and umbilical cord length (UCL). A retrospective cohort study was performed on aborted or prematurely delivered stillborn fetuses submitted for necropsy, and a follow-up questionnaire survey was sent to mare owners to obtain additional anamnestic data. In total 72 cases were included in the study. Multivariable regression analyses were performed to evaluate the risk factors (mare size, period of gestation, gestational length, parity, previous abortion, UCL, maternal age at abortion and fetal sex) association with UCT and UCL, and Wilcoxon rank sum test tested pairwise differences in UCL for the risk factors mare size, period of gestation and cause of abortion. Abortion caused by UCT (UCTA) was diagnosed in 37 cases, and these had shorter gestational length (8.1 ± 0.3 months) compared to fetuses lost to other causes (8.9 ± 0.2 months) (p = 0.002). UCTAs accounted for the highest prevalence of abortions in the mid gestational period (113–227 days) (14/21, 67%) compared to the late period (228–314 days) (22/50, 44%) (p = 0.05). The cause of abortion was associated with period of gestation (p = 0.009) and UCL (p = 0.002), and fetuses aborted as a consequence of UCT had longer UCs (98.84 ± 5.14 cm) compared to abortions due to other causes (67.2 ± 4.9 cm). Mares in the mid period of gestation had 7.9 times the odds of UCTA compared to the late period (CI: 1.93, 32.2), and UCL was associated with increased odds of UCTA (OR: 1.05, CI: 1.02, 1.07). UCL was associated with mare size (p = 0.012), period of gestation (p = 0.002) and cause of abortion (p = 0.001). In conclusion, UCTA was significantly associated with UCL and gestational length. The prevalence of UCTA was not influenced by mare size, but pony fetuses from UCTAs had significantly shorter UCs compared to horse fetuses. UCs longer than 54 cm may predispose ponies for UCTA. No other maternal or fetal risk factors were found to be associated with the cause of abortion.

Congenital malformations caused by Akabane virus in porcine fetuses in southern Japan.

Akabane virus (AKAV) is known as a major teratogenic agent of ruminant fetuses. In this study, we investigated the relationship between porcine abnormal deliveries and AKAV by serology, pathology, and virology investigations using specimens from 16 stillborn fetuses delivered in southern Japan between 2013 and 2015. The major clinical manifestations in stillborn fetuses were hydranencephaly, arthrogryposis, spinal curvature, and both skeletal muscle and subcutaneous edema. Histologic examination of the specimens identified atrophy of skeletal muscle fibers accompanied by adipose replacement. Nonsuppurative encephalomyelitis and decreased neuronal density in the ventral horn of the spinal cord were shown in two separate fetuses, respectively. Neutralizing antibody titers to AKAV were detected in most of the tested fetuses (13/16). The AKAV sequences detected in the affected fetuses in 2013 and 2015 were highly identical and closely related to Japanese AKAV isolates which were isolated in 2013 and sorted into genogroup I of AKAV. Immunohistochemistry visualized AKAV antigens in the neuronal cells of the central nervous system of the fetuses. These findings indicate that AKAV was involved in the birth of abnormal piglets at the affected farm. The clinical manifestations and histopathological features in the stillborn fetuses were very similar to those in ruminant neonates affected by AKAV. To avoid misdiagnosis and to evaluate the precise impact of AKAV on pig reproduction, AKAV should be considered in differential diagnoses of reproductive failures in pigs.

CYP26B1-related disorder: expanding the ends of the spectrum through clinical and molecular evidence.

CYP26B1 metabolizes retinoic acid in the developing embryo to regulate its levels. A limited number of individuals with pathogenic variants in CYP26B1 have been documented with a varied phenotypic spectrum, spanning from a severe manifestation involving skull anomalies, craniosynostosis, encephalocele, radio-humeral fusion, oligodactyly, and a narrow thorax, to a milder presentation characterized by craniosynostosis, restricted radio-humeral joint mobility, hearing loss, and intellectual disability. Here, we report two families with CYP26B1-related phenotypes and describe the data obtained from functional studies of the variants. Exome and Sanger sequencing were used for variant identification in family 1 and family 2, respectively. Family 1 reflects a mild phenotype, which includes craniofacial dysmorphism with brachycephaly (without craniosynostosis), arachnodactyly, reduced radioulnar joint movement, conductive hearing loss, learning disability-and compound heterozygous CYP26B1 variants: (p.[(Pro118Leu)];[(Arg234Gln)]) were found. In family 2, a stillborn fetus presented a lethal phenotype with spina bifida occulta, hydrocephalus, poor skeletal mineralization, synostosis, limb defects, and a synonymous homozygous variant in CYP26B1: c.1083C > A. A minigene assay revealed that the synonymous variant created a new splice site, removing part of exon 5 (p.Val361_Asp382del). Enzymatic activity was assessed using a luciferase assay, demonstrating a notable reduction in exogenous retinoic acid metabolism for the variant p.Val361_Asp382del. (~ 3.5 × decrease compared to wild-type); comparatively, the variants p.(Pro118Leu) and p.(Arg234Gln) demonstrated a partial loss of metabolism (1.7× and 2.3× reduction, respectively). A proximity-dependent biotin identification assay reaffirmed previously reported ER-resident protein interactions. Additional work into these interactions is critical to determine if CYP26B1 is involved with other biological events on the ER. Immunofluorescence assay suggests that mutant CYP26B1 is still localized in the endoplasmic reticulum. These results indicate that novel pathogenic variants in CYP26B1 result in varying levels of enzymatic activity that impact retinoic acid metabolism and relate to the distinct phenotypes observed.

Placental inflammation and pregnancy outcomes: A prospective, observational study in South Asia: The PURPOSe study.

To examine inflammatory lesions in placentas of stillbirths, preterm neonatal deaths and term controls in India and Pakistan. Prospective, observational study. Three hospitals in India and a large maternity hospital in Pakistan. The enrolled participants with placentas available for histology evaluation included stillbirths (n = 814), preterm live births who died within 28 days of birth (n = 618) and term live birth controls (n = 201). From this same population, polymerase chain reaction (PCR) analysis for pathogens was performed on 809 stillbirth placentas, 614 neonatal death placentas and the placentas of 201 term controls. Placentas from preterm infants who lived beyond day 28 (n = 1432) were only available from India. A prospective observational study of placental inflammatory lesions defined by the Amsterdam criteria and on the same placentas, multiplex PCR evaluation for 75 pathogens using TaqMan Array Cards. Any placental inflammatory lesions, including chorioamnionitis, funisitis, villitis and intervillitis and their association with various pathogens. In the Indian liveborn preterm infants, placental inflammation of any kind was present in 26.2% of those who died versus 16.6% of those who lived (p = 0.0002). Chorioamnionitis was present in 25.8% of those who died versus 16.3% of those who lived (p = 0.0002) and funisitis was present in 4.1% of those who died versus 1.5% of those who lived, (p = 0.005). Across all three sites, in the placentas of the 201 term controls, 18.9% had any inflammation, 16.9% had chorioamnionitis, 5.5% had funisitis, 0.5% had intervillitis and none had villitis. Overall, for stillbirths, any inflammation was observed in 30.2%, chorioamnionitis in 26.9%, funisitis in 5.7%, intervillitis in 6.0% and villitis in 2.2%. For the neonatal deaths, any inflammation was present in 24.9%, chorioamnionitis in 23.3%, funisitis in 8.1%, intervillitis in 1.9% and villitis in 0.5%. Compared with the placentas of term controls, in neonatal deaths, only chorioamnionitis was significantly increased (23.3% versus 16.9%, p = 0.05). Among stillbirths, the rates of any inflammation, chorioamnionitis, intervillitis and villitis were similar across the birthweight groups. However, funisitis was more common in the placentas of stillborn fetuses weighing 2500 g or more (13.8%) compared with 1.0% for those weighing less than 1000 g and 4.8% for stillborn fetuses weighing 1000-2499 g. In the PCR studies, Ureaplasma spp. were by far the most common pathogens found and generally were more commonly found in association with inflammatory lesions. Chorioamnionitis was the most common type of placental inflammatory lesion regardless of whether the placentas evaluated were from term controls, stillbirths or neonatal deaths. For stillbirths, inflammation in each inflammation category was more common than in the term controls and significantly more so for any inflammation, chorioamnionitis, intervillitis and villitis. For neonatal deaths, compared with the placentas of term controls, all inflammation categories were more common, but only significantly so for chorioamnionitis. Ureaplasma spp. were the most common organisms found in the placentas and were significantly associated with inflammation.

An Immunohistochemical Study of Proliferation of Human Fetal Heart Cardiomyocyte With Phospho-Histone H3 Antibody.

The rapid proliferation of cardiomyocytes in mammals occurs during fetal life. But in postnatal life, this capacity of proliferation is reduced or lost as they exit the cell cycle. However, the cardiomyocytes don't show the same activity for different species. In human fetuses orin adult life, the capacity of the proliferation of cardiomyocytes and their response to an injury arenot understood yet.In this study, we have done an immunohistochemical study using phospho-histone H3 (PHH3) to observe human fetal cardiomyocytes' proliferative activity. The heart specimens from the fetal autopsy of spontaneously aborted and stillborn human fetuses were subjected to immunohistochemical study using PHH3 antibody, and comparison between the PHH3 index (number of PHH3 positive cells per 1000 number of cardiomyocytes/high power field [HPF]) of myocardial regions was done using appropriate statistical tests. A total of 17 fetal hearts were included in our study. In the left ventricle, right ventricle, right atrium, and interventricular septum, the PHH3 index of myocardium was significantly higher over the pericardial region (p-value 0.002, p-value <0.001, <0.001, and 0.009 respectively) as compared to the region of over the endocardium and the middle part of the myocardium.The PHH3 index of the pericardial region of the left ventricle was significantly correlated with the maximum thickness of the left ventricle.