COVID-19 during the Third Semester of Pregnancy: Maternal Characteristic, Possibility of Intrauterine Transmission and Neonatal Outcome in Aceh, Indonesia

BACKGROUND: Pregnant women are vulnerable against COVID-19 infection due to physiological and immunological changes. COVID-19 in pregnancy affects fetal well-being with a potential for vertical infection. AIM: This study aims to determine the incidence of vertical infection and anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies in infants born to mothers with positive COVID-19 infection. MATERIALS AND METHODS: Amniotic fluid, swabs of the newborn's nasopharynx and oropharynx, and swabs of the placenta were examined using reverse transcription polymerase chain reaction (RT-PCR) for SARS-CoV-2. Serological examination was performed by Electro-Chemiluminescence Immunoassay on infant's blood. RESULT(S): Four of 33 pregnant women gave birth to infants positive SARS-CoV-2 infection. RT-PCR examination of all amniotic fluid and placental swabs was negative for SARS-CoV-2. Four of 33 infants (12.1%) showed negative polymerase chain reaction (PCR) results but positive SARS-CoV-2 antibodies, another 4 newborns (12.1%) showed positive PCR results, but no SARS-CoV-2 antibodies detected. The remaining 25 babies (75.8%) showed both negative PCR and serologic results. CONCLUSION(S): No evidence of vertical transmission found in this study.Copyright © 2023 Cut Meurah Yeni, Zinatul Hayati, Sarjani M. Ali, Hasanuddin Hasanuddin, Rusnaidi Rusnaidi, Cut Rika Maharani.

Perinatal aspects of sars-cov-2 infection during pregnancy: A potential cause for concern

Ever since the beginning of COVID-19 pandemic, uncertainty regarding clinical presentation and differences among various subpopulations exist. With more than 209,870,000 confirmed cases and more than 4,400,000 deaths worldwide, we are facing the new era of health crisis which will undoubtedly impair global health, economic and social circumstances. In the past year, numerous genetic mutations which code SARS-CoV-2 proteins led to the occurrence of new viral strains, with higher transmission rates. Apart from the implementation of vaccination, the effect of SARS-CoV-2 on pregnancy outcome and maternal fetal transmission remains an important concern. Although neonates diagnosed with COVID-19 were mostly asymptomatic or presented with mild disease, the effect on early pregnancy is yet to be evident. While positive finding of SARS-CoV-2 RNA in some samples such as amniotic fluid, placental tissue, cord blood and breast milk exists, additional research should confirm its association with transplacental transmission.Copyright © 2022, Dr. Mladen Stojanovic University Hospital. All rights reserved.

Exosomes/EVs: IMMUNOMODULATORY EFFECTS OF AMNITOIC FLUID EXTRACELLULAR VESICLES IN AN IN-VITRO MODEL OF IMMUNE ACTIVATION

Background & Aim: Amniotic fluid (AF)-derived EVs are currently under investigation for use as anti-inflammatory therapeutics in COVID-19 and COVID-19 long haulers. The dysregulation of the immune response induced by SARS-COV-2 is a key driver of both acute COVID-19 induced lung injury and long term COVID-19 sequela. There is a clear need to identify therapeutics that suppress excessive inflammation and reduce immune cell exhaustion to improve patient short term and long-term outcomes. Amniotic fluid (AF)- derived extracellular vesicles (EVs) have previously been shown to deliver anti-inflammatory and immune-modulatory signals to diverse cellular targets. We aimed to test if AF-EVs carry immune-suppressive molecules and can suppress T-cell immune activation and exhaustion in vitro. Methods, Results & Conclusion(s): The AF-EV biologic tested was derived from AF collected from consenting donors during planned, fullterm cesarean sections. AF was centrifuged and filtered to remove cellular debris and create a product containing AF-EVs and soluble extracellular components. Fluorescent EXODOT analysis was performed to demonstrate the presence of EV markers CD9, CD81, ALIX, and immune suppressive molecule PD-L1. T-cell activation/exhaustion was induced in vitro by treating human peripheral blood mononuclear cells with activation agent PHA for 3 days with the addition of AF-EVs or saline control. Immune activation/exhaustion was measured by flow cytometry to determine the expression of PD-1 on CD3+ T-cells. The AF-EV biologic was characterized to contain EVs with positive expression of CD9, CD81, ALIX, and PL-L1. T-cell activation/exhaustion was upregulated in response to PHA and was significantly reduced by 8% in AF-EV treated T-cells compared to saline control (77.7% vs 85.7%, respectively P<0.05). These findings demonstrate that AF-EVs do express PD-L1, a surface marker that has previously been demonstrated to contribute to exosome-mediated immunosuppression. Furthermore, we confirmed in vitro that AF-EVs suppress T-cell activation/ exhaustion in the presence of a T-cell activation agent. COVID-19 long haulers have been described to have upregulated and pro-longed immune activation and T-cell exhaustion, marked by an increase in PD1+ T-cells. Therefore, this finding serves as a starting point for the development of a potential mechanism of action that may describe AF-EV's therapeutic effect in COVID-19 long hauler patients.Copyright © 2023 International Society for Cell & Gene Therapy

Evolution in Infectious Causes of Death in Liveborn Children in a Major Tertiary Children's Hospital: A Retrospective Autopsy Review of over 1200 Cases

Background: Infections have historically been a leading cause of death, particularly in children. Medical advances, including vaccines and antimicrobials, have significantly decreased infection-related deaths, but infections remain a cause of pediatric mortality, especially in premature infants. The types of infections implicated in childhood deaths have changed with these advances, for example, meningitis and meningococcal infections were leading causes in 1981 but not in the later period. The incidence and etiologies of infection- related deaths may be altered by major events that modify not only medical practices but also societal attitudes and activities. Examples of such events include the HIV/AIDS epidemic that began in the early 1980s and the more recent COVID-19 pandemic. In order to investigate changes in infection-related pediatric deaths over time, we analyzed and compared autopsy cases performed during 5-year span prior to both the HIV/AIDS epidemic and the COVID-19 pandemic in which infections contributed to death. Method(s): Review of all autopsy cases performed at our institution between 1/1/1975-1/1/1980 and between 1/1/2015-1/1/2020 was performed to identify cases in which infection directly contributed to death, comprising 1262 cases. Only liveborn children were considered, and neonatal sepsis from amniotic sac infections was excluded. Comparison of decedent characteristics and infectious etiologies between the two time periods was performed, identifying age, race, sex, gestational age (for decedents less than 3 months of age), and etiologic class of agent (bacterial, viral, fungal or parasitic). TORCH infections and vaccine-preventable illnesses were specifically assessed. Proportions were compared using 1 (assessing TORCH, vaccine-preventable, and prematurity deaths)- or 2-tailed (all others) z-tests, with significance calculated at the < 0.05 level. Result(s): In the 1970s cohort, 300 infectious autopsy cases were identified in liveborn children;73 were identified in the 2010s. Compared to the 2010s cohort, the 1970s decedents were more likely to be white (85% v 53%, p=0.012), comprise children aged 1-5 and 13+ (22% v 6.8% [p=0.003] and 16.4% v 8.3% [p=0.036]), and were less likely to be premature (66.7% v 80.4%, p=0.039). Vaccine-preventable illnesses (for example: measles) accounted for 36 deaths in the 1970s cohort but only 2 in the 2010s cohort (p=0.009). Thirteen children died of TORCH infections (CMV, toxoplasmosis and HSV) versus 5 in the 2010s (CMV and HSV), which did not reach statistical significance. Conclusion(s): Pediatric mortality secondary to infections has decreased significantly compared to fifty years ago, especially in younger children and in relation to vaccine-preventable infections such as meningococcal disease. This drop is largely attributed to medical advances, including vaccines and antimicrobial medications. Additional contributing factors could include practices adopted post-HIV/AIDS, especially in the community. Further exploration of how such changes in medical and social practice impacted mortality and comparing them to changes occurring in the intra/post-COVID-19 era, is helpful. Yet, with the increased survival of premature infants, they remain at risk of devastating consequences from infections.

Placental Thrombosis Resulting in Hypoxic Ischemic Encephalopathy in an Infant of a Mother with Covid-19

Case Report: Since the beginning of the Coronavirus Disease 2019 (COVID-19) pandemic, there has been much work to understand the negative effects of SARS-CoV-2 on tissues expressing the Angiotensin Converting Enzyme-2 (ACE2) receptor, including the placenta. However, there is limited information regarding placental pathology findings in mothers with COVID-19 and the effects of SARS-CoV-2 on the placenta. The available research reports effects on the fetus ranging from minimal to intrauterine fetal demise. Case Description: A 4680g baby boy was born at 38+1 weeks of gestation to 36y old G4P1021 female via repeat cesarian section. The pregnancy was complicated by advanced maternal age, chronic hypertension with superimposed pre-eclampsia with severe features, BMI of 80, and SARS-CoV-2 infection. The mother had mild COVID-19 symptoms and did not require hospitalization or oxygen support. Prenatal ultrasounds were limited due to body habitus. At the time of delivery, there was clear amniotic fluid. Upon delivery the infant was cyanotic and limp and was brought to the warmer immediately. Non-invasive positive pressure ventilation was initiated at 5 minutes of life with improvement in infant color and oxygen saturation. He was then admitted to the Neonatal Intensive Care Unit (NICU). APGARs were 2, 3, 5, and 7 at 1, 5, 10, and 15 minutes respectively. Cord gases showed severe metabolic acidosis. The patient was diagnosed with hypoxic-ischemic encephalopathy (HIE) and therapeutic hypothermia was initiated. Both the NICU and obstetric teams were unable to identify a clear perinatal cause of HIE in this patient. Later, the placenta pathology report revealed a large placenta for estimated gestational age corresponding to the 75th percentile, villous parenchyma with focal chorangiosis and thrombi, with unremarkable fetal membrane and three vessel umbilical cord. The cause of HIE was then thought to be due to the placental thrombi likely caused by SARS-CoV-2 infection. Discussion(s): Fetal vascular malperfusion and fetal vascular thrombus have been noted as a common finding in the placentas of pregnant women who test positive for SARS-CoV-2. There are various causes of HIE, from maternal, placental and fetal factors. This patient had no clinically evident hypoxic event, but information was limited due to the lack of monitoring of the fetus in utero. Given the mother's SARS-CoV-2 infection and the placental pathology findings, it is likely that the cause of this patient's HIE was related to the effects on the placenta from SARS-CoV-2. Conclusion(s): As more information comes to light about the effects of SARS-CoV-2 on the placenta, it is important to consider a maternal SARS-CoV-2 infection during pregnancy as a cause of HIE in a newborn.

Epidemiological study of COVID-19 pneumonia in pregnant woman and their neonates, report of thirteen confirmed COVID-19 pregnant women

Background: In new pandemic, the probable effects of COVID-19 pneumonia on pregnant woman and their infant is one of new critical challenge for health care. Here we presented clinical symptoms, laboratory findings and outcome of COVID-19 pneumonia in pregnant woman. Method(s): In a case series study, from 15 Feb to 15 June 2020, all women with RT-PCR COVID-19 who referred to two hospitals (Taleghani and Qods Hospital) affiliated to Arak University of Medical Sciences were selected. The epidemiological and demographic variables, laboratory test and outcomes obtained from patient's medical records. Result(s): In this case series, we presented thirteen confirmed COVID-19 pregnant women. Their mean age was 34.6 (S.D.: 5.9) years and the mean gestational age was 32.4 (S.D.: 7.3) weeks. Most of patient didn't show any maternal complication and intrauterine vertical transmission. The large number of pregnant women had normal HRCT and also in terms of laboratory most of the patients had normal laboratory tests. Amniotic fluids, cord blood, the throat swab of neonate in our pregnant woman with delivery were tested for COVID-19 and all of them were negative. Conclusion(s): The COVID-19 mothers and their infant didn't have higher risk for morbidity and mortality and this virus didn't associate with intrauterine vertical transmission.Copyright © 2020 Ubiquity Press. All rights reserved.

Presence of SARS-CoV-2 neutralizing and vaccine spike antibodies in amniotic fluid during the second trimester of pregnancy, detected by amniocentesis (Winner of the SIMP Award, in memory of Professor Giorgio Pardi)

Objective. Thanks to the SARS-CoV-2 vaccination, pregnant women are protected from the complications of COVID-19 infection, but the benefits of this vaccination in preventing morbidity and mortality in the fetus are not yet clear: it is not well understood if and how these antibodies cross the placenta. Indeed antibodies made after a pregnant person has received an mRNA COVID-19 vaccine have been found in amniotic fluid and umbilical cord blood at term and represent a safer method of enhancing neonatal antibody levels than administration of immunoglobulin preparation to the infant. The aim of the study is to test the presence of neutralizing SARS-CoV-2 antibodies and spike antibodies in the amniotic fluid in the second trimester of pregnancy, and then to compare the antibodies level in maternal serum and amniotic fluid to evaluate their correlation. Materials and Methods. This cohort study took place at the Department of Obstetrics and Gynecology of Messina at the AOU Policlinico G. Martino from September 2021 to February 2022;the study consisted of 22 pregnant women who had amniocentesis in the gestational period between 15 weeks plus 6 days and 18 weeks: we analyzed serum and amniotic fluid samples of women who contracted the SARS-CoV-2 infection, or who were vaccinated against the same virus, within one year, or never infected by SARS-CoV-2 or vaccinated against it. During the amniocentesis, all patients underwent a single sample of maternal serum and of amniotic fluid to evaluate SARS-CoV-2 neutralizing antibody and S1 receptor binding domain IgG antibody levels. Inclusion criteria were pregnant women with the need to undergo amniocentesis. Results. 22 pregnant women were enrolled in the study:10 of them were vaccinated with a mRNA COVID-19 vaccine;12 women were not vaccinated, 4 of them had developed COVID-19 infection within one year before the collection and 2 of them developed the infection during pregnancy;the other 6 never developed the infection and have not been vaccinated, enrolled as comparators. Mann-Whitney test showed that vaccinated patients had significantly higher S1 receptor binding domain antibody levels both in amniotic fluid (p < 0.006) and maternal blood (p < 0.005) than not vaccinated women;also SARS-CoV-2 neutralizing antibody levels were higher in pregnant women who developed COVID-19 infection both in amniotic fluid (p < 0.007) and maternal blood (p < 0.004) than not vaccinated women. There was a significantly high correlation between the concentrations of spikes antibody levels in vaccinated pregnant women's serum and amniotic fluid (p = 0.000), and of neutralizing antibody levels in serum and amniotic fluid of women who developed COVID-19 infection (p = 0.000). Conclusions. To the best of our knowledge, the analysis of amniotic fluid and serum showed for the first time that all the vaccinated pregnant women samples had SARS-CoV-2 spikes immunoglobulins both in maternal blood and amniotic fluid. There is a very high correlation between maternal blood and amniotic fluid S1 receptor binding domain antibody levels in vaccinated women: this demonstrates that there is an early transplacental antibody transfer. Also neutralizing antibodies were found in the amniotic fluid of infected pregnant women, with high correlation between concentrations.

Aplasia cutis congenita in dizygotic twin infants

Aplasia cutis congenita (ACC) is often sporadic, but familial cases have been reported. We report a case of a dichorionic diamniotic twin pregnancy in which both the male and female twins had matching areas of aplasia cutis on their scalps. An Irish couple sought fertility treatment using a donor egg and paternal sperm. Successful in vitro fertilization (IVF) and the transfer of two embryos resulted in a diamniotic dichorionic twin pregnancy. Two fetal poles were noted at the 12-week ultrasound (US) scans. The mother suffered from a minor urinary tract infection during the first trimester but had no other history of infection, including herpes simplex virus or COVID- 19. She was known to be varicella immune prior to pregnancy. The twins were born by elective caesarean section owing to breech presentation. Twin one was female and twin two was male. Both infants were born with scarring on the crown of their head, which was consistent with ACC. Cranial US showed no underlying bony abnormality. The rest of the cutaneous examination was normal and there were no other congenital anomalies. ACC is a rare, heterogeneous group of disorders characterized by the congenital absence of skin, which can be focal or widespread. It is thought to affect 1-3 per 10 000 live births. The exact cause of ACC is unclear. Various hypotheses have been suggested, including defective closure of the neural tube or embryonic fusion lines, intrauterine trauma, placental insufficiency, fetus papyraceus, amniotic membrane adhesions, intrauterine infections, teratogens and genetic mutations. The classification of ACC is based on the area affected, type of skin irregularity, associated congenital defects and mode of inheritance. Scalp ACC without multiple anomalies (category 1) is generally associated with an autosomal dominant or sporadic pattern of inheritance. These twins may have an autosomal dominant mutation that led to this phenotype. ACC can also be associated with fetus papyraceus or placental infarct. This is less likely in this case as only two embryos were transferred, and the pregnancy was dichorionic. Most cases of ACC associated with fetus papyraceus occur in monozygotic pregnancies. ACC lesions often heal spontaneously by re-epithelialization resulting in a hairless superficial scar. Twin one had a slightly smaller area affected by ACC and overlying eschar resolved several weeks after birth. Twin two has had no hair growth in the area. This case highlights the difficulties in ascertaining the aetiology of this rare condition in twin pregnancies.

Detection of Breast Milk and Amniotic Fluid in Late Pregnant Women with Asymptomatic Infection of Coronavirus Disease 2019: Case Report

It was to explore the clinical characteristics of late pregnant women with asymptomatic infection of coronavirus disease 2019 (COVID-19) and the risk of intrauterine vertical transmission and breastfeeding transmission. The clinical data of a late pregnant woman with COVID-19 were retrospectively analyzed. The patient (singleton, cephalic) was admitted to the hospital on April 3, 2022. The patient had a dramatic increase in C-reactive protein after delivery, other blood routine tests were unremarkable, and the result of nucleic acid test was positive. On April 13, 2022, due to oligohydramnios, under measures such as positive pressure headgear and protective clothing in negative pressure operating room, second cesarean section combined with hysterorrhaphy (emergency) was performed to terminate the pregnancy, and the mother and baby were in good condition. Postpartum amniotic fluid, milk, and neonatal nucleic acid tests were negative. Corresponding treatment was given after cesarean section, and the patient was continuously monitored for nucleic acid detection to meet the discharge criteria of the COVID-19 and discharged and isolated. After discharge, the patient was followed up for 1 month. The newborn's spirit, diet, and development were normal. The maternal cesarean section incision recovered well. Patient with COVID-19 in the third trimester has prolonged hospital stays and are difficult to treat. However, the virus was not found in amniotic fluid, breast milk, and newborn, for which COVID-19 does not lead to intrauterine vertical transmission and breastfeeding transmission in the third trimester.Copyright © 2023 by the authors.

COVID-19 pandemic and neonatal health: What we know so far?

COVID-19 first time appeared in December 2019 in Wuhan, China. The number of cases increased rapidly in china and outside and the World Health Organization declared a pandemic on 11th March 2020. The pregnant and postpartum women, child, and neonatal populations are vulnerable to this disease due to immunological and physiological changes. This paper analyzed the published evidence for assessing the effect of COVID-19 on neonatal health and health care. Online published literature was searched from PubMed, Google Scholar, and other official webpages using keywords: "coronavirus/COVID-19/new coronavirus 2019"/SARS-CoV-2 and neonatal health/care/outcomes" and reviewed to prepare this article. COVID-19 is the potential to transmit either mother to fetus or mother/caregiver to neonates. However, neonates born from infected mothers did not show significant clinical features. Pharyngeal-swab, amniotic-fluid, cord-blood, and breast-milk test results were not found positive. Health facility-based vaginal/caesarian delivery was considered a low risk of transmission. However, recommended to separate neonates with infected mothers/caregivers and test immediately after birth to avoid the possible transmission. Mothers/caregivers should take routine preventive measures such as washing hands frequently and avoiding contact with infected people. If neonates suffered from the server acute respiratory distress requires intensive care urgently. Despite the possibility of the intrauterine transmission of COVID-19 direct evidence is still lacking so it needs more studies for further confirmation. The International Pediatric Association suggested preventive programs, curative care, vaccination, and telemedicine care as the minimum services and called on its members to address these cares during the pandemic. Copyright © 2020, Kathmandu University. All rights reserved.

Histopathological finding of 5 cases of CAOS without genital bleeding

Objective: Chronic Abruption Oligohydramnios Sequence (CAOS) is known as a disease with a poor perinatal prognosis characterized by placental marginal hematoma and genital bleeding, associated with fetal growth restriction, hemorrhagic amniotic fluid, oligohydramnios and chronic lung disease in the newborn. We reviewed the placental pathology of five cases diagnosed as CAOS without genital bleeding. Method(s): Pregnant women with CAOS without genital bleeding between April 2020 and July 2022 were identified. Five of these cases were clinically suspected as CAOS. Therefore, we evaluate the presence of hemorrhagic amniotic fluid and/or posterior placental hematoma by MRI;one case could not perform MRI because of COVID-19 infection. Placentas were examined histopathological and reviewed for the four Amsterdam classification groups, presence of diffuse chorioamniotic hemosiderosis (DCH), and other special findings. Result(s): The median age of the patients was 31 years and the median number of weeks of delivery was 24 weeks, including 2 cases of artificial abortion, 1 case of vaginal delivery, and 2 cases of cesarean section. The main complaints were FGR in 2 cases, amniotic fluid depletion in 2 cases, and suspected fetal congenital anomaly in 1 case. In histopathologic examination, 4 cases show DCH, 3 cases show maternal vascular marperfusion, and all cases had Acute chorioamnionitis. Conclusion(s): Four of the five patients had DCH, which suggest that the chorion and amnion are exposed to hematoma over a long period of time. This finding provides the basis for a final diagnosis in cases of clinically suspected CAOS. Our cases suggested that CAOS may be established before the appearance of genital hemorrhage. MRI or other tests should be performed aggressively in cases of clinically suspected CAOS without genital hemorrhage, and the patient should be strictly managed. Copyright © 2022

An RT-PCR Assay for Detection of Infectious Bronchitis Coronavirus Serotypes

Avian infectious bronchitis virus (IBV), a chicken Gammacoronavirus, is a major poultry pathogen, and is probably endemic in all regions with intensive poultry production. Since IBV was first described in 1936, many serotypes and variants of IBV have been isolated worldwide. IBV isolates are capable of infecting a large range of epithelial surfaces of the chicken, involving the respiratory, renal, and reproductive systems;however, the clinical signs are usually not specific for differential diagnoses. Virus isolation is commonly used for diagnosis of IBV infection, which was achieved through passage of clinical materials via the allantoic route of embryos. Currently, more sensitive molecular approaches for the detection of avian pathogens have been developed, including reverse-transcriptase polymerase chain reaction (RT-PCR) and real-time RT-PCR, which are more suitable for use in diagnostic laboratories. In this chapter, we describe a one-step RT-PCR which can be used for detecting most of IBV serotypes in the IBV-infected allantoic fluid and has been used routinely in our laboratories for detection of IBVs. Copyright © Springer Science+Business Media New York 2016.

Covid-19 infection mimicking postpartum pulmonary embolism: A case report

The case was a 32-year-old, nulliparous pregnant woman, after in-vitro fertilization (IVF) pregnancy. Her contractions started, and the amniotic fluid membrane ruptured at 38 gestational ages. The patient developed shortness of breath and lower oxygen saturation which started suddenly at the 8th hour after the cesarean section procedure under general anesthesia. Thrombotic conditions that may appear during the postpartum period should be diagnosed early and immediate treatment should be started to resolve the actual cause. Copyright © 2022 Ondokuz Mayis Universitesi. All rights reserved.

Covid 19 with hematological presentation and complications that may lead to stem cell transplant

Background: This pandemic has impacted health and the economy worldwide on an unprecedented scale. It has important systemic effects including the cardiovascular and immune systems. Aim(s): Discuss the links between COVID-19 clinical features and complications and its hematological findings and coagulopathy. This link may shed light on our prognostic view to patients with COVID-19 and will have significant therapeutic implications. One of these important implications is the use of mesenchymal stem cells (MSCs) to treat COVID-19 patients. Method(s): Review done to Autopsy findings support the concept that the pathogenesis of severe COVID-19 involves direct viral-induced injury of multiple organs, including heart and lungs, coupled with the consequences of a procoagulant state with coagulopathy. Mesenchymal stem cells as a potential therapy for COVID-19 INFECTION. Result(s): Mesenchymal stem cells (MSCs) can be isolated from different adult tissues, and neonatal birth-associated tissues, including placenta (PL), umbilical cord (UC), Warton jelly (WJ), amniotic fluid (AF), and cord blood (CB), and then stored for future possible applications. Differentiation potential, powerful immunoregulation, and endogenous repair mechanisms. Support to potential use of MSCs in treating patients came from observations that MSCs have been identified to efficiently cure ALI/ARDS from both infectious and noninfectious causes, mediated primarily by paracrine mechanisms based on the released extracellular vesicles (EVs). The immune-regulation of MSCs depends mainly on modulating activation and effector function of immune cells, suppressing lung-infiltrated cells, and enhancing the resolution of pulmonary edema [35]. Conclusion(s): COVID-19 is associated with distinct hematological changes, rise in serum inflammatory markers, and coagulopathy. Most of these changes were related to the patients' prognosis and mortality, particularly in those with severe disease. There are links between COVID-19 clinical features and complications and its hematological findings and coagulopathy. COVID-19 patients seems to be a promising approach. Further studies are needed to clarify more clinic-pathologic links in COVID-19 that might improve outcomes.

Protective Action of Phosphorus 6CH in SARS-CoV-2 Spike Protein Induced Pathogenecity in Gallus-gallus Embryo

Introduction: In the search of effective medicines against Coronavirus Disease-2019 (COVID-19) besides the conventional mode of treatment many medicines belonging to alternative therapeutics claimed to be effective in this disease. In homeopathy-a branch of alternative medicine some medicines are claimed to be effective in COVID-19 after human trials. Aim: To study whether ultradiluted preparation of Phosphorus 6CH (centesimal (C) dilutions, using Hanhemann's (H) dilution method) can protect damaging action of Delta Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) spike protein Receptor Binding Domain (RBD) in Gallus gallus embryo in relation to their gross appearances, histopathological changes and cytokine changes. Materials and Methods: An in-vivo fertilized chick embryo model experimental analysis was carried out at the Genetic Research Laboratory of Heritage Institute of Technology, Kolkata, West Bengal, India. The whole experimental study was done in a time period of November 2021 to January 2022 and the data collected were analysed using statistical software Minitab. About 14 days old Gallus gallus embryonated eggs were inoculated with the antigen along with the vehicle alcohol controls. The Phosphorus 6CH was used to see whether it can prevent or cure the damaging action of the spike protein in the embryo in different experimental sets. results: The notable finding in this experiment is the remarkable elevated expression of Interleukin (IL)-10 gene in the curative, preventive sets as well as in the medicine control sets in comparison to antigen and alcohol control sets. In case of Transforming Growth Factor, (TGF) β1 there was enhanced expression of TGF β1 gene in the alcohol 6C set and antigen set which gets ameliorated with Phosphorus 6CH. The morbid anatomy of the embryo and the histopathological picture of the liver of the embryo also reflected similar findings in these two experimental sets. After statistical analysis it was found that there was significant correlation in between Interferon (IF) γ and IL-10 in these experimental results which appears very important. conclusion: The homeopathic medicine phosphorus 6CH is capable of maintaining cytokine balance in Delta SARS-CoV-2 spike protein RBD induced pathogenecity in Gallus gallus embryo.

Exosomes/EVs: Late Breaking Abstract: EFFECTS OF AMNIOTIC FLUID DERIVED EXTRACELLULAR VESICLE BIOLOGIC ON T CELL PROLIFERATION AND ACTIVATION

Background & Aim: Extracellular Vesicles (EVs) isolated from single cell sources such as mesenchymal stem cells and biological fluids such as amniotic fluid have previously been found to have various effects on immune cell functions. Zofin is an acellular biologic derived from amniotic fluid and contains naturally occurring EVs. Zofin is currently under clinical investigation for COVID-19 infection and Post Acute Sequale of COVID-19. Therefore, we aimed to investigate the effects of Zofin on the lymphocyte immune response in an in vitro cell culture model. Methods, Results & Conclusion: Zofin was manufactured from fullterm amniotic fluid collected from healthy, consenting mothers during planned cesarean section. The Zofin used in these experiments contained 1.5x1011 particles/ml as quantified by Nanosight NS300. To study the immune response in vitro, PBMCs were stimulated with 5μg/ml Phytohemagglutinin (PHA) for up to 8 days. PBMCs were labeled before culture with cell trace dye to monitor changes in proliferation. Stimulated PBMCs were treated with either serum-free media (vehicle) or different concentrations of Zofin (10% and 20%). 4 days following PHA stimulation, Zofin suppressed PHA-induced T cell proliferation in a dose-dependent manner. 8 days following PHA stimulation there were higher rates of proliferation, Zofin maintained suppressed proliferation at 20% concentrations. In the same cell culture model, treatment with Zofin attenuated a PHA-induced increase in T cell activation as observed by a reduction in expression of activation markers. Overall, this data demonstrates that Zofin has an effect on the immune response of T cells and suggests amniotic fluid derived EVs could be used as an immunotherapeutic tool

Exosomes/EVs: QUANTIFICATION OF CD81+ AND CD133+ EXTRACELLULAR VESICLE POPULATIONS DERIVED FROM AMNIOTIC FLUID

Background & Aim: Amniotic fluid (AF)-derived extracellular vesicles (EVs) are currently being studied within clinical trials as a novel therapeutic drug for acute and chronic diseases such as COVID-19, osteoarthritis, and chronic obstructive pulmonary disease (COPD). Recently, AF has been characterized to contain harvestable EVs that reduce cytokine expression and protect from high oxygen tissue injury. However, the reproducibility of EV populations derived from AF samples has yet to be fully explored. Here within, we present results of a fluorescent nanoparticle tracking analysis that characterizes the reproducibility of CD81+ and CD133+ populations of EVs. Methods, Results & Conclusion: AF was collected from consenting adults during planned, full term cesarean sections and then processed via filtration and ultracentrifugation to precipitate the nanoparticle population. Prior to fluorescent analysis, a MACSPlex exosome kit (Miltenyi Biotec) was used to identify potential exosome surface markers. Nine independent samples of AF-derived nanoparticles were used for Zetaview analysis. Each pellet was resuspended in saline and stained with CD133-AF488 and CD81-DyLight55 antibodies. Fluorescent nanoparticle tracking analysis was completed using the Zetaview Quatt (Particle Metrix). Subsequent videos of stained and total particles were recorded to determine the percentage of positive stained nanoparticles. Exosome surface marker analysis revealed 10 out of the 37 tested surface markers to be present with the most intense surface markers being CD81 and CD133. Furthermore, detection of CD81+ and CD133+ nanoparticles via Zetaview was found in all samples (n=9). The AF- derived nanoparticles were CD81+ and CD133+ at 73.51 ± 18.35 % and 30.41 ± 14.06 % (Mean ± SD), respectively. A high percentage of the nanoparticles were CD81+ with a smaller yet apparent display of CD133+ nanoparticles throughout the samples. Fluorescent characterization of the AF-derived samples demonstrated the reproducibility of both CD81+ and CD133+ nanoparticles. Studying the nature of these nanoparticle populations helps researchers to characterize samples consistently and could lead to understanding their therapeutic potential as a novel biologic.

Umbilical cord vascular thrombosis: literature review and two clinical cases

Thrombosis of the umbilical cord vessels is a rare complication of pregnancy, combined with a high level of perinatal morbidity and mortality. Anomalies of vascular attachment (velamentous attachment), pathology of the umbilical cord (hyperspiralization, short or long umbilical cord), intrauterine infections, maternal diabetes mellitus and preeclampsia as well as meconium found in the amniotic fluid are among the risk factors of developing thrombosis in the umbilical cord vessels. Here we present two clinical observations of umbilical vein thrombosis at full-term pregnancy. In both cases, during pregnancy and childbirth, no signs of umbilical cord pathology were found according to cardiotocography and Doppler ultrasound;despite this, the children were born in hypoxic state. Both newborns were transferred to the second stage of treatment due to suspected intrauterine pneumonia. Velamentous attachment, intrauterine infections as well as meconium found in the amniotic fluid were the risk factors of developing umbilical vein thrombosis described in case 1 and case 2, respectively. During pregnancy, both female patients suffered from clinically confirmed novel coronavirus infection (COVID-19) and contacted patients with COVID-19 in the third trimester of gestation. It is likely that endothelial damage caused by the novel coronavirus SARS-CoV-2 was one of the risk factors for the development of umbilical vein thrombosis, but this issue requires to be further explored.

Frequency of Meconium-stained Amniotic Fluid in COVID-positive Term Pregnancies and Perinatal Outcome

Aim: To know the frequency of meconium-stained amniotic fluid (MSAF) among COVID-positive term pregnant mothers and to know perinatal outcomes in these neonates. Materials and methods: A retrospective study was conducted of COVID-positive term pregnancies admitted to Vanivilas hospital, Bangalore Medical College, during the study period of 1 year. The study period was from 1st of July 2020 to 31st of June 2021. High-risk pregnancies that confound the occurrence of MSAF were excluded (obstetric risk factors and medical risk factors complicating pregnancy). Data were collected from medical records of patients and parturition register, which are cross-verified with operation theater records and neonatal intensive care unit (NICU) registers. Results: In total, 200 pregnant women were included in the study. About 65% were delivered by cesarean, 34.5% by the vaginal route, and one vacuum-assisted delivery. In total, 199 live births and one fresh stillbirth are reported. About, 26% had MSAF and 6.5% of newborns had meconium aspiration syndrome. Grade I, II, and III cases were 4.5%, 12.5%, and 9% respectively. Low appearance, pulse, grimace, activity, and respiration (APGAR) scores were correlating with the grade of MSAF, which is statistically significant. There were two neonatal deaths in grade III MSAF cases. Conclusion: The frequency of MSAF is increased among COVID-positive mothers, which translates to low APGAR scores and poor perinatal outcomes. Vigilant intrapartum care is recommended for these pregnancies to reduce the risk of poor neonatal outcomes. Clinical significance: The current study is undertaken to know if coronavirus disease-2019 (COVID-19) is associated with an increased frequency of MSAF among COVID-positive pregnant women. Finding MSAF has implications on operative delivery, cesarean rates, and perinatal outcome. There is a lack of studies about MSAF and perinatal outcomes in COVID-positive mothers. Evidence generated by this study helps to counsel the COVID-positive mothers and guides in the management of COVID-positive laboring women.

Study of Pregnant Women Infected with COVID-19 During Pregnancy: A Case Report

This case study includes three pregnant women with COVID-19 diagnosed during pregnancy or delivery between March 28 and May 13, 2020. All cases were confirmed by a positive pharyngeal reverse transcription polymerase chain reaction (RT-PCR) test and one case by computed tomography scan (CT Scan) in addition to the (RT-PCR). Clinical and laboratory information was extracted from hospital records during pregnancy and delivery. The adverse effects during pregnancy and after the birth of the newborn, the possibility of vertical transmission from positive pregnant mothers to the neonates were investigated.Of the three women with COVID-19 infection, one patient was diagnosed two weeks before delivery and two were diagnosed during delivery and hospitalization. No adverse effects including preeclampsia, gestational hypertension, rupture of the amniotic sac during pregnancy and premature delivery were observed but one of the patients suffered from intrauterine fetal death (IUFD). in this study, adverse pregnancy outcome was not observed in pregnant women with Covid-19 infection based on hospital observations. No vertical transmission was observed following vaginal delivery or cesarean section and during pregnancy. As the effect of the virus on different people in society varies according to their individual characteristics, our conclusion in this study on pregnant women is also affected by these individual differences, which requires further studies in this field with more samples.