20th Day Of Gestation Research Articles

Effects of Prenatal Methcathinone Exposure on the Neurological Behavior of Adult Offspring.

Our previous research has shown that prenatal methcathinone exposure affects the neurodevelopment and neurobehavior of adolescent offspring, but the study on whether these findings continue into adulthood is limited. This study aims to explore the effects of prenatal methcathinone exposure on anxiety-like behavior, learning and memory abilities, as well as serum 5-hydroxytryptamine and dopamine concentrations in adult offspring. Pregnant rats were injected daily with methcathinone between the 7th and 20th days of gestation. The neurobehavioral performance of both male and female adult offspring rats was evaluated by neurobehavioral tests, including open-field tests, Morris water maze (MWM) tests, and novel object recognition (NOR) tests. The levels of 5-hydroxytryptamine and dopamine concentration in rat serum were detected by ELISA. Significant differences were found in the length of center distance and time of center duration in the open-field test, as well as the times of crossing the platform in the MWM test, between the prenatal methcathinone exposure group and the control group. Results of the NOR test showed that adult offspring rats exposed to methcathinone need more time to discriminate the novel object. No gender differences were detected in the neurobehavioral tests. The serum concentrations of 5-hydroxytryptamine and dopamine in rats exposed to methcathinone prenatally were lower than that in the control group, and the serum dopamine concentration was independent of gender in each group. Prenatal methcathinone exposure affects the neurological behavior in adult offspring, and 5-hydroxytryptamine and dopamine might be involved in the process.

Effects of apelin on neonatal brain neurogenesis in L-NAME-induced maternal preeclampsia

The aim of this study was to investigate the possible protective effects of apelin, which is known to have antioxidant and anti-inflammatory effects, on changes in neurogenesis in newborns of pregnant rats with L-NAME-induced preeclampsia. Wistar albino female rats were divided into four experimental groups: Control, Apelin, Preeclampsia and Preeclampsia + Apelin. Blood pressure was measured on the 5th, 11th and 17th days of gestation, urine protein was analyzed from urine samples collected for 24 h on the 6th, 12th and 18th days and serum creatinine was analyzed from serum samples. Maternal kidney and placenta tissues were obtained to establish the preeclampsia model, and neonatal brain tissues including the cortex, hippocampus and cerebellum regions were obtained to investigate neurogenesis and examined by histological and immunohistochemical methods. The number of newborns, body weight and brain weight of the newborns were measured. eNOS, IL-10, nNOS and NO levels in the brain analyzed via ELISA. Mean arterial pressure, urine protein and serum creatinine increased in the preeclampsia. Newborn weight decreased in the Preeclampsia group, the values in the Preeclampsia + Apelin group were closer to the Control and Apelin groups. In the Preeclampsia group, edema and dilatation in the proximal and distal tubules of kidneys, perivillous fibrin deposition and increase in syncytial nodules of placenta were observed. VEGF immunoreactivity decreased and iNOS immunoreactivity increased in both kidney and placenta. In neonatal brain tissue examinations, cytotoxic edema accompanied by thinning of cortex, delayed migration and lower cell counts in the hippocampus, and increase in intercellular spaces and EGL thickening in the cerebellum were observed in the preeclampsia. Expression of NeuN, GFAP, MBP, IL-10, eNOS, nNOS and NO levels decreased, whereas expression of Iba-1 increased in the preeclampsia. In the Preeclampsia + Apelin group, these findings were similar to the Control and Apelin groups. Apelin administration was found to be beneficial for preventing the adverse consequences of preeclampsia, but further experimental and clinical studies are needed to better understand these effects.

Manganese porphyrin-based treatment improves fetal-placental development and protects against oxidative damage and NLRP3 inflammasome activation in a rat maternal hypothyroidism model

Oxidative stress (OS) and endoplasmic reticulum stress (ERS) are at the genesis of placental disorders observed in preeclampsia, intrauterine growth restriction, and maternal hypothyroidism. In this regard, cationic manganese porphyrins (MnPs) comprise potent redox-active therapeutics of high antioxidant and anti-inflammatory potential, which have not been evaluated in metabolic gestational diseases yet. This study evaluated the therapeutic potential of two MnPs, [MnTE-2-PyP]5+ (MnP I) and [MnT(5-Br-3-E-Py)P]5+ (MnP II), in the fetal-placental dysfunction of hypothyroid rats. Hypothyroidism was induced by administration of 6-Propyl-2-thiouracil (PTU) and treatment with MnPs I and II 0.1 mg/kg/day started on the 8th day of gestation (DG). The fetal and placental development, and protein and/or mRNA expression of antioxidant mediators (SOD1, CAT, GPx1), hypoxia (HIF1α), oxidative damage (8-OHdG, MDA), ERS (GRP78 and CHOP), immunological (TNFα, IL-6, IL-10, IL-1β, IL-18, NLRP3, Caspase1, Gasdermin D) and angiogenic (VEGF) were evaluated in the placenta and decidua on the 18th DG using immunohistochemistry and qPCR. ROS and peroxynitrite (PRX) were quantified by fluorometric assay, while enzyme activities of SOD, GST, and catalase were evaluated by colorimetric assay. MnPs I and II increased fetal body mass in hypothyroid rats, and MnP I increased fetal organ mass. MnPs restored the junctional zone morphology in hypothyroid rats and increased placental vascularization. MnPs blocked the increase of OS and ERS mediators caused by hypothyroidism, showing similar levels of expression of HIFα, 8-OHdG, MDA, Gpx1, GRP78, and Chop to the control. Moreover, MnPs I and/or II increased the protein expression of SOD1, Cat, and GPx1 and restored the expression of IL10, Nlrp3, and Caspase1 in the decidua and/or placenta. However, MnPs did not restore the low placental enzyme activity of SOD, CAT, and GST caused by hypothyroidism, while increased the decidual and placental protein expression of TNFα. The results show that treatment with MnPs improves the fetal-placental development and the placental inflammatory state of hypothyroid rats and protects against oxidative stress and reticular stress caused by hypothyroidism at the maternal-fetal interface.

Effect of curcumin on inflammatory and oxidative stress markers in Lipopolysaccharide induced animal models of pre-eclampsia

Introduction: A substantial percentage of fetal, maternal, and neonatal morbidity and mortality is traced to Pre-eclampsia, a life threatening disease. Typical symptoms of the disease include de novo hypertension, proteinuria, renal failure, neurological problems, elevated liver enzymes, and uteroplacental dysfunction. It is estimated that almost 8-10% of all pregnancies are affected by preeclampsia in India. Pre-eclampsia incidentally also involves immune system changes on similar lines as a typical pregnancy. Immune cells like CD4+ T-helper 1 (TH1) cells, cytotoxic NK cells, and autoreactive B cells secrete substances that cause an increase in innate immune activation and inflammation in the maternal circulation and uteroplacental unit, instead of immune changes that promote tolerance and inhibit reactivity to the fetus and placenta. Placental and systemic oxidative stress are the key factors in the emergence of preeclampsia. A number of physiological changes, mineral shortages, and increased oxygen use during pregnancy can all be considered potential causes of oxidative stress. There is yet no effective and secure pharmaceutical treatment for preeclampsia. Pregnancy related safety concerns and the complex aetiology are two significant barriers to the development of pre-eclampsia medications. Medicinal plants and derived products have long been recognized as valuable assets in the treatment of a variety of diseases. Curcumin is a naturally occurring substance with significant in vitro therapeutic promise. The objective of this study was to develop LPS (lipopolysaccharide) induced animal model of Pre-eclampsia and to estimate and compare the hemodynamic parameters, inflammatory markers, oxidative stress markers and proteinuria in pregnant control, LPS-induced Pre-eclampsia, and curcumin-treated Pre-eclampsia in experimental rats. Material and methods: Pre-eclampsia was induced by single dose of 0.5μg/kg LPS. Wistar rats were divided into 8 groups of 8 animals. Female wistar rats were placed with male rats overnight and examined the next morning for the presence of sperm in vaginal smears. The day on which sperm was detected was designated as gestational day 0. After confirmation of pregnancy, each rat was weighed and randomized into groups. Pregnant Wistar rats were given 0.5 μg /kg of body weight LPS intraperitoneally on gestational day 5 for developing Pre-eclampsia models. Protein estimation in urine was done after 24 hours. On 19th or 20th gestation day animals were anesthetized and blood pressure was measured through a non-invasive tail-cuff method using the AD instrument and also through invasive method by inserting a arterial catheter in either of the carotid arteries towards the heart. ECG recording was done through surface ECG recording. Blood was collected by cardiac puncture, centrifuged, and stored at -80ºC on 19th or 20th gestation day. Inflammatory markers were estimated through ELISA reader. Oxidative stress markers were estimated. Pregnancy outcome was assessed through placental weight, number of pup’s delivered and pup’s weight. Statistical analysis was done using one way analysis of variance (ANOVA) followed by post hoc Tukey‘s test for intergroup comparisons. Results: In LPS induced model of pre-eclampsia, single-dose treatment with LPS (0.5ug/kg) induced marked pre-eclampsia in rats. Treatment with curcumin significantly decreased the level of proteinuria, hemodynamic parameters, inflammatory markers and oxidative stress markers in pregnant LPS+ curcumin-treated group when compared with the pregnant LPS group. Treatment with curcumin significantly increases pup’s weight and number of pups. Conclusion: On the basis of this experimental study, it is proposed that curcumin appears to be a potential therapeutic agent as measured by effect on hemodynamic changes, proteinuria, inflammatory markers, oxidative stress markers and pregnancy outcomes in LPS-induced pre- eclampsia. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Modulatory effects of quercetin on histological changes, biochemical and oxidative stress of rat placenta induced by inhalation exposure to crude oil vapor

The inhalation exposure to crude oil vapor (COV) has been shown to have adverse effects on the placenta and fetal development. The modulatory effects of quercetin (QUE) as a natural phenolic compound with antioxidant properties are promising for the protection of placental structure. This study aimed to investigate the modulatory role of QUE in mitigating histopathological damage, oxidative stress, and biochemical alteration in the placenta of COV-exposed pregnant rats. Forty-eight pregnant rats were divided into eight groups (days 15 and 20) as follows: 1–2) Control groups, 3–4) COV groups, 5–6) COV+QUE groups, and 7–8) QUE-treated groups (50 mg/kg). The inhalation method was used to expose pregnant rats to COV, and QUE was administered orally. On the 15th and 20th days of gestation, placental tissue was analyzed using PAS and H&E staining and immunohistochemistry. The expression of the caspase-3 gene and oxidative stress biomarkers including TAC, CAT, MDA, GPx, and SOD were investigated in the placental tissue. The COV significantly decreased the weight, diameter, and thickness of the placenta as well as the thickness of the junctional zone and labyrinth and the number of trophoblast giant cells in 15- and 20-day-old placentas (P<0.05). Also, COV significantly increased placental expression of caspase-3 and the oxidative stress biomarkers (P<0.05). The administration of QUE along with exposure to COV reduced morphometric and histological alteration, oxidative stress, and caspase-3 expression (P<0.05). Our findings indicated that QUE in COV-exposed pregnant rats can prevent placental histopathological alternations by increasing the activity of the antioxidant system.

Behavioral, Maternal and Neurological Effects of Monosodium Glutamate on Pregnant Female Rats During Early and Late Gestation

Seventy-five adult Wistar Albino rats were used in this experiment to detect the effect of MSG on fetus morphology, and pregnant rats’ behavioral, biochemecal, and neurological parameters. They were housed under standard laboratory conditions in plastic cages in a quiet environment with free access to food and water. They were randomly assigned into 4 groups of three replicates for each. Group (1) control, received normal saline. Groups (2, 3, and 4) received daily MSG orally at a dose of 1, 3 and 6 mg/kg respectively for a period of 20 days. There was a reduction in the mean number of fetuses /dam, an altered distribution of implantation, reduction in fetuses’ weight &amp; length, besides some fetuses revealed morphological anomalies and skeletal defects at the 19th day of gestation. In addition, there was a decrease in the number of entrance and spontaneous alternations, increase in line crossing and rearing behaviors, elevation in serum ALT, AlP and creatinine, decrease in serum progesterone, increase in dopamine and glutamate levels (ng/mL) in brain tissues of MSG treated pregnant rats. So that, this study suggests that to diminish MSG in foods during pregnancy to to stay away from harmful effects on pregnant females behavior, brain tissue, and safe fetuses from deformity.

Embryo and Fetal Toxic Effects of the Hydroethanol Extract of Urtica simensis Hochst. Ex. A. Rich Leaves in Pregnant Rats

Introduction: Urtica simensis has been used to treat various diseases such as malaria, hypertension, diabetes, gonorrhea, gastritis, body swelling, and wound infections. However, the safety of consuming U. simensis leaves during pregnancy has not been evaluated yet. Therefore, this experimental study was conducted to evaluate the toxic effects of U. simensis leaf extract on the prenatal development of embryos and fetuses in pregnant rats.Methods: Fifty pregnant Wistar albino rats were randomly assigned to five groups of 10 gravid rats for each experiment. Groups I–III were given 70% ethanol leaf extract of U. simensis at doses of 250, 500, and 1000 mg/kg daily from 6th to 12th days of gestation. Groups IV–V were kept as pair‐fed and ad libitum controls. The developing embryos and fetuses were retrieved on 12 days and 20 days of gestation, respectively. Embryos were evaluated for growth and developmental delays. Fetuses were also assessed for growth retardation and external and visceral anomalies.Results: In the embryonic experiment, somite numbers (p = 0.001) and morphological scores (p = 0.029) were significantly decreased in pregnant rats given 1000 mg/kg of U. simensis leaf extract. Embryonic developments of the caudal neural tube (CNT) (p = 0.001), otic system (p = 0.025), olfactory system (p = 0.013), and limb buds (p = 0.026) were significantly delayed in pregnant rats given 1000 mg/kg of extract. Oral administration of 500 mg/kg of U. simensis leaf extract also caused significant developmental delays in the CNT (p = 0.021) and olfactory system (p = 0.032). In the fetal experiment, fetal resorption (p = 0.015) was significantly increased whereas crown rump length (p = 0.012) and fetal weight (p = 0.019) were significantly decreased in pregnant rats given 1000 mg/kg of U. simensis leaf extract.Conclusions: The embryotoxic effects of U. simensis leaf extract were evidenced by significant developmental delays. The fetal toxic effects of U. simensis leaf extract were also shown by significant decreases in fetal growth indices. Therefore, pregnant women should be well informed of the possible toxic effects of consuming U. simensis leaf during pregnancy.

Antagonistic Effects of Chinese Salt and Folic Acid on Developing Swiss Albino Mice

One of the most often utilized taste enhancer in commercial meals is monosodium glutamate (MSG) commonly called Chinese salt. MSG utilization has been increasing over time and linked with toxicity in liver and other organs. Objective: To determine the teratogenic and toxic effects of Chinese salt and folic acid on developing mice. Methods: In this study, 20 female pregnant albino mice were divided into four groups, each consisted 5 mice. The control group was supplied with water. To check the teratogenicity and toxicity of Chinese salt and folic acid the treated groups such as Group-I, Group-II and Group-III were supplied with Chinese salt and/or folic acid of concentration 7.50 ug/g of their body weight. The dose was administered orally on daily basis during 6th to 12th day of gestation. This was accomplished through an 18 days trial. On the eighteenth day of gestation, the pregnant mice underwent dissection and the fetuses were retrieved. Fetuses were taken from all groups for histopathological findings and morphometry. Results: Vast range of morphological, morphometric and histological abnormalities in mice were observed in pregnant mice and fetuses. Conclusions: The findings of this study clearly revealed that Chinese salt and folic acid overdose are potentially toxic to liver and stomach.

DIA-based quantitative proteomic analysis of porcine endometrium in the peri-implantation phase

The 12th day of gestation is a critical period for embryo loss and the beginning of imminent implantation in sows. Data independent acquisition (DIA) technology is one of the high-throughput, high-resolution and reproducible proteomics technologies for large-scale digital qualitative and quantitative research. The aim of this study was to identify and characterize the protein abundance landscape of Yorkshire pig endometrium on the 12th day of pregnancy (P12) and estrous cycle (C12) using DIA proteomics. A total of 1251 differentially abundant proteins (DAPs) were identified, of which 882 were up-regulated and 369 were down-regulated at P12. Functional enrichment analysis showed that the identified proteins were related to metabolism, biosynthesis and signaling pathways. Three proteins were selected for Western blot (WB) validation and the results were consistent with the DIA data. Further combined with transcriptome data, fibrinogen like 2 (FGL2) and S100 calcium binding protein A8 (S100A8) were verified to be highly abundant in the P12 endometrial epithelium. In summary, there were significantly different abundance of proteome profiles in C12 and P12 endometrium, suggesting that DAPs are associated with changes in endometrial receptivity, which laid the foundation for further research on related regulatory mechanisms. SignificanceThe 12th day of gestation is an important point in the peri-implantation period of pigs, when the endometrium presents a receptive state under the stimulation of estrogen. DIA proteomics technology is an emerging protein identification technology in recent years, which can obtain protein information through comprehensive and unbiased scanning. In this study, DIA technology was used to characterize endometrial proteins in pigs during the peri-implantation period. The results showed that higher protein abundance was detected using the DIA technique, and some of these DAPs may be involved in regulating embryo implantation. This study will help to better reveal the related proteins involved in embryo implantation, and lay a foundation for further research on the mechanism of endometrial regulation of embryo implantation. Significance of the studyThe 12th day of gestation is an important point in the peri-implantation period of pigs, when the endometrium presents a receptive state under the stimulation of estrogen. DIA proteomics technology is an emerging protein identification technology in recent years, which can obtain protein information through comprehensive and unbiased scanning. In this study, DIA technology was used to characterize endometrial proteins in pigs during the peri-implantation period. The results showed that higher protein abundance was detected using the DIA technique, and some of these DAPs may be involved in regulating embryo implantation. This study will help to better reveal the related proteins involved in embryo implantation, and lay a foundation for further research on the mechanism of endometrial regulation of embryo implantation.

Long-term consequences of acute toxic prenatal hypoxia induced by sodium nitrite in rat experiments

Introduction. Gestational hypoxia plays an important role in fetal development, as it causes the development of oxidative stress and delayed psychomotor development. The aim of the study was to evaluate the prenatal effect of sodium nitrite on indices characterizing orienting and exploratory activity, mental activity and the degree of hypoxia in rat offspring. Materials and methods. Acute hypoxia was modeled by administration of sodium nitrite at a dose of 30 mg/kg intramuscularly to five pregnant rats during 16–19 days of gestation. Five females constituted a control group. In 50 born rats, orientation and exploration activity, cognitive functions, intensity of lipid peroxidation and glycolysis were studied. Results. The offspring of females that underwent acute hypoxia significantly lagged behind in the positive dynamics of body weight gain compared to control individuals. Thirty-day-old rats showed a pronounced disorder of the structure of behavioral pattern, learning process and long-term memory, increased content of diene conjugates, malonic dialdehyde and the level of lactate dehydrogenase activity in blood. Discussion. It was found that offspring whose mothers suffered acute hypoxia of toxic genesis developed pronounced behavioral disorders of psychomotor agitation and cognitive dysfunction characterized by decreased learning ability and impaired long-term memory. The content of primary and secondary products of lipid peroxidation and lactate dehydrogenase was increased in rats. The development of cellular oxidative stress under conditions of intrauterine hypoxia leads to oxygen deficiency in the fetus and a switch from tissue respiration to glycolysis, which underlies CNS damage. Conclusion. It was shown that sodium nitrite administration during the period from the 16th to the 19th day of gestation caused hyperactivity, depressed emotional status, and decreased orienting and exploratory activity and the ability to learn and retain a memory trace in rat offspring. Increased levels of diene conjugates and malonic dialdehyde, lactate dehydrogenase activity were observed in rats.

The effect of HSP70 overexpression on the cerebral cortex development

Heat shock proteins (HSPs) make up a large family of molecular chaperones, recognized for their role in protein maturation, refolding, and degradation. HSP70 was shown to promote cell survival during several pathological processes in the brain, such as stroke, neurodegenerative diseases, epilepsy, and trauma [1]. In addition, HSPs serve to promote the proper embryonic and postnatal development of many organ systems, such as the central nervous system [2]. Heat shock proteins demonstrate specific expression patterns throughout the development of the nervous system, notably during crucial embryonic and postnatal moments [1]. During embryonic development, neural and glial progenitors must survive within a hypoxic microenvironment while performing energetically demanding actions, such as cell migration and neurite outgrowth. HSPs can activate or inhibit development pathways in the nervous system that modulate cell differentiation, neurite outgrowth, cell migration, or angiogenesis [1]. Indeed, recent studies demonstrated that HSP70 directly regulates the development of the nervous system by modulating signaling cascades involved in cell growth and migration [3]. Additionally, research demonstrated [4] that introducing HSP70 from an external source significantly augments the populations of proliferating cells and differentiated neuroblasts within the mouse hippocampus. Nevertheless, some researchers [1] contend that overexpression of HSPs may negatively impact cell survival. Therefore, the precise role of these chaperones remains largely unexplored. In our study, we used in utero electroporation to introduce plasmids that controlled HSP70 overexpression into neuron progenitors of mouse embryos on the 14th day of gestation. Additionally, plasmid DNA encoding GFP was used to facilitate subsequent visualization of transformed cells. Brain samples were collected on the 18th day of gestation for immunohistochemical analysis of the sections. Confocal microscopy was used to compare the characteristics of neuronal migration in both control and HSP70 overexpression conditions. Cells that received plasmids inducing HSP70 were discovered to migrate at a lower pace in comparison to the control. Additionally, it is hypothesized that the induction of HSP70 expression could lead to neuronal malformations and impact the development of neurites. In the future, the study of cytoarchitectonics in the cortex will continue, examining the identification of electroporated cells in individual neuron populations using Satb2 and Ctip2 markers. Additionally, the differentiation of these cells will be assessed by counting those that have not exited the mitotic cycle, and the hypothesis of apoptosis induction in cells electroporated with HSP70 will be tested.

Prenatal developmental sequences of the esophageal epithelium in the New Zealand white rabbits: Light and electron microscopic analysis.

Several morphogenetic sequences occur during esophageal development and birth defects occur due to defects in foregut morphogenesis. This work aimed to record the cellular events in the morphogenesis of rabbits' esophageal epithelium. On the 16th day of gestation, the esophageal epithelium varied from stratified ciliated columnar to stratified squamous type. The surface epithelium presented mucous cells with mucigen granules of various sizes occupying their supranuclear cytoplasm. Cytoplasmic vacuolation was evident in all layers of the esophageal epithelium at this age. On the 18th gestational day, some light cells could be detected in the middle portion of the epithelium, while others occupied the whole epithelial length. On the 21st day, mucous cells are more frequently observed at the apical esophageal part as well as at the surface epithelium. Numerous elongated dark cells could be distinguished embedded between the basal cells. On the 24th gestational day the number of the mucous cells reached its peak. Reaching the 30th gestational day, several lamellar bodies, a keratinized layer and mitotic divisions could be demonstrated, and the number of both mucous and dark cells was greatly decreased. Collectively, detection of surface mucous and dark cells together with the non-cornified surface in some regions of the rabbit esophageal epithelium at the end of gestation ensure a postnatal development to reach the adult epithelium essential to sustain the passage of the harsh raw food. Future immunohistochemical studies are recommended to investigate the components of secretions in mucous cells and functional studies to highlight the dark cells significance. RESEARCH HIGHLIGHTS: Esophageal epithelium of fetal rabbit was analyzed by light and transmission microscopy. Surface epithelium presented mucous cells with mucigen granules of various sizes. They reached their maximum number on 24th day then decreased. On the 16th day, cytoplasmic vacuolation was evident in all epithelial layers. On the 21st day, numerous elongated dark cells could be distinguished embedded between the basal cells. Before birth, several lamellar bodies, a keratinized layer and mitotic divisions could be demonstrated, and the number of both mucous and dark cells was greatly decreased.

Matrix Metalloproteinase-20 immunolocalization in rat first molar tooth development after treatment with amoxicillin

Amoxicillin is one of the most commonly prescribed antibiotics in children. It is, therefore, administered as the first choice of antibiotics for respiratory, gastrointestinal, neuronal, and skin infections. This study aimed to determine whether amoxicillin use affects the formation of dentin and enamel during the secretion and early phases of mineralization. Sixteen pregnant adult Wistar rats were equally divided into two groups. The first group that did not receive the drug was prescribed a saline solution (control group), and the other group received 250 mg/kg/day amoxicillin (study group). The treatments were administered daily by oral gavage from the 13th gestation day to the end of gestation. After birth, the newborn also received the same treatment as their mothers from the first day of birth until 7 or 12 days after birth. The newborns will be sacrificed at 7 and 12 days postnatally. The jaws will be dissected, and the maxilla, where the samples are fixed in 10% formaldehyde solution, and the upper first molars will be analyzed. Immunostaining with MMP-20 on day 7 and 12 were performed. A significant difference in the positive expression of ameloblast for MMP20 antibody between groups was observed at 7 days (P = 0.014) and no significant difference at 12 days (P = 0.347). A significant difference for positive expression of odontoblast for MMP20 antibody between groups in both durations was observed at 7 days (P = 0.178) and not a significant difference at 12 days (P = 0.143) and powerful expression in enamel matrix during early mineralization stage in the control group and strong expression in the study group. The current findings indicate that amoxicillin affects the expression of MMP20 during the secretory stage by decreasing the expression of MMP20 in ameloblast, and the expression of MMP20 decrease in cells during the early mineralization stage in both groups at different rate. Keywords: Amoxicillin, Enamel, Immunostaining, MMP20, ameloblast

Teratoma induced by amlodipine drug in fetus rats

The current study, performed during the period from February 2021 to June 2022 at the University of Thi- Qar/ College of Education for Pure Sciences, and aimed to follow the changes in external morphological features at different Embryonic developmental stages in pregnant rats treated with different doses of amlodipine. Usage In this study, 18 pregnant rats were randomly divided into three groups, each with six pregnant rats. Each group received different concentrations of amlodipine (0.3, 0.5) in oral doses until the 20th day of gestation, while the control group was injected with 0.9% normal saline. Teratomas are bizarre tumors derived from embryonic tissue that are normally found only in the gonadal and sacral regions of adults. Primary retroperitoneal teratomas are rare and present challenging management options. In one outcome, a teratoma was found in one of the amlodipine-treated groups. Complete excision of the teratoma mass was performed.

G-protein-coupled receptor MAS deletion produces a preeclampsia-like phenotype in FVB/N mice.

An unbalance in the renin-angiotensin (Ang) system (RAS) between the Ang II/AT1 and Ang-(1-7)/Mas axis appears to be involved in preeclampsia (PE), in which a reduction in Ang-(1-7) was observed. Here, we tested whether the reduction in the activity of the Ang-(1-7)/Mas axis could be a contributing factor for the development of PE, using Mas-deficient (Mas-/-) mice. Cardiovascular parameters were evaluated by telemetry before, during pregnancy and 4 days postpartum in 20-week-old Mas-/- and wild-type (WT) female mice. Mas-/- mice presented reduced arterial blood pressure (BP) at baseline (91.3 ± 0.8 in Mas-/- vs. 94.0 ± 0.9 mmHg in WT, Diastolic, P<0.05). However, after the 13th day of gestation, BP in Mas-/- mice started to increase, time-dependently, and at day 19 of pregnancy, these animals presented a higher BP in comparison with WT group (90.5 ± 0.7 in Mas-/- vs. 80.3 ± 3.5 mmHg in WT, Diastolic D19, P<0.0001). Moreover, pregnant Mas-/- mice presented fetal growth restriction, increase in urinary protein excretion as compared with nonpregnant Mas-/-, oliguria, increase in cytokines, endothelial dysfunction and reduced ACE, AT1R, ACE2, ET-1A, and eNOS placental mRNA, similar to some of the clinical manifestations found in the development of PE. These results show that Mas-deletion produces a PE-like state in FVB/N mice.

Micronuclei and nuclear buds in amniotic tissue of rats treated with cyclophosphamide.

Fetal development can be altered by DNA damage caused by maternal exposure to chemical, physical, or biological agents during gestation. One method of assessing genotoxicity is to detect micronuclei (MNs) and/or nuclear abnormalities. This can be performed in vivo and requires only frequently dividing tissues, such as amniotic tissue (AT), which is in contact with the fetal environment and is composed of very thin layers of cells. This study evaluated the presence of MNs, nucleoplasmic bridges, and nuclear buds (NBs) in the fetal AT following maternal exposure to cyclophosphamide (CP) during pregnancy. Pregnant Wistar rats were divided into a negative control group and an experimental group that was orally administered CP (10mg/kg). Daily blood smears were obtained from pregnant rats on days 14-19 of gestation. The rats were dissected, and fetal ATs were obtained on the 19th day of gestation. The MN and NB frequencies in AT cells were analyzed using a fluorescence microscope (100×). Micronucleated erythrocytes in the peripheral blood of the control rats were also assessed. Micronucleated polychromatic erythrocyte frequencies were significantly higher than those in the controls. Polychromatic erythrocyte frequencies were lower in CP-treated rats than in controls at 48-120h. Fetuses in the CP-treated group also showed a significant increase in MNs and NBs in AT cells. In conclusion, AT could be used for analyzing MNs and NBs in rats following maternal exposure to a genotoxic agent and as a viable alternative for analyzing the integrity of fetal DNA during gestation.

Identification and characterization of circRNAs in peri-implantation endometrium between Yorkshire and Erhualian pigs

BackgroundOne of the most critical periods for the loss of pig embryos is the 12th day of gestation when implantation begins. Recent studies have shown that non-coding RNAs (ncRNAs) play important regulatory roles during pregnancy. Circular RNAs (circRNAs) are a kind of ubiquitously expressed ncRNAs that can directly regulate the binding proteins or regulate the expression of target genes by adsorbing micro RNAs (miRNA).ResultsWe used the Illumina Novaseq6,000 technology to analyze the circRNA expression profile in the endometrium of three Erhualian (EH12) and three Yorkshire (YK12) pigs on day 12 of gestation. Overall, a total of 22,108 circRNAs were identified. Of these, 4051 circRNAs were specific to EH12 and 5889 circRNAs were specific to YK12, indicating a high level of breed specificity. Further analysis showed that there were 641 significant differentially expressed circRNAs (SDEcircRNAs) in EH12 compared with YK12 (FDR < 0.05). Functional enrichment of differential circRNA host genes revealed many pathways and genes associated with reproduction and regulation of embryo development. Network analysis of circRNA-miRNA interactions further supported the idea that circRNAs act as sponges for miRNAs to regulate gene expression. The prediction of differential circRNA binding proteins further explored the potential regulatory pathways of circRNAs. Analysis of SDEcircRNAs suggested a possible reason for the difference in embryo survival between the two breeds at the peri-implantation stage.ConclusionsTogether, these data suggest that circRNAs are abundantly expressed in the endometrium during the peri-implantation period in pigs and are important regulators of related genes. The results of this study will help to further understand the differences in molecular pathways between the two breeds during the critical implantation period of pregnancy, and will help to provide insight into the molecular mechanisms that contribute to the establishment of pregnancy and embryo loss in pigs.

Human Wharton's jelly-derived mesenchymal stem cells prevent pregnancy loss in a rat by JAK/STAT-mediated immunomodulation.

Spontaneous abortion (SA) is a multiple-original syndrome with immune imbalance as one of its major risk factors. As Wharton's jelly-mesenchymal stem cells (WJ-MSCs) are considered to be able to prevent abortion, this study aims to explore the currently poorly understood underlining molecular signaling pathways and regulatory mechanisms of WJ-MSCs in pregnancy maintenance. Abortion mode is established by subcutaneous injection of bromocriptine in rat on day 9 and abortion prevention is achieved by WJ-MSCs injection via tail vein. WJ-MSCs were cultured with/without the inhibitors of JAK/STAT or NF-κB. The uterus was collected on the 14th day of gestation and the rate of embryo absorption was calculated. The expression of Th1/Th2/Th3 cytokines in decidual, placental tissue, and peripheral blood was analyzed. WJ-MSCs treatment significantly reduced the abortion rate in bromocriptine-treated pregnancy such that it was not significantly different from a normal pregnancy. JAK/STAT inhibition abolished pregnancy preserving effects of WJ-MSCs but NF-κB inhibition did not. The levels of Th1-related cytokines and mRNA levels in the bromocriptine abortion model were significantly higher than the normal pregnancy group and ethanol control group, while levels of the Th2-related cytokines and mRNA levels significantly decreased. WJ-MSCs transfusion into the abortion model restored cytokine profiles such that they were not significantly different from the normal pregnancy group and ethanol control group. JAK/STAT inhibition of WJ-MSCs prevented their effect on cytokine and mRNA levels, but NF-κB inhibition did not. WJ-MSCs significantly lower the rate of embryo resorption of spontaneous abortion by reducing Th1-related cytokines while increasing Th2 and Th3-related cytokines in JAK/STAT-dependent manner.

Protective effect of Agaricus bisporus mushroom against maternal and fetal damage induced by lead administration during pregnancy in rats.

Lead (Pb) is a toxic pollutant, which can affect different tissues of the human body. The use of natural elements, as medicinal mushroom can reduce the toxic effects of Pb. We evaluated, through preclinical tests, the oral co exposures to mushroom Agaricus bisporus (Ab) by gavage and Pb in drinking water, and the capability of Ab be a protective agent for both pregnant rats and their fetuses. Female Wistar rats were divided into four groups (n = 5/group): Group I-Control; Group II-Ab 100 mg/kg; Group III-Pb 100 mg/L; Group IV-Ab +Pb -100 mg/kg +100 mg/L. Exposure was performed until the 19th day of gestation. On the 20th day, pregnant rats were euthanized, and the outcomes evaluated were weight gain; hematological profile; biochemical markers; oxidative stress markers; reproductive capacity; and embryo fetal development. The characterization of mushrooms reveals them to be a valuable source of nutrients. However, Pb ingestion resulted in reduced weight gain and negative impacts on hematological and biochemical parameters. Fortunately, co administration of mushrooms helped to mitigate these negative effects and promote recovery. The mushroom also showed antioxidant activity, improving parameters of oxidative stress. In addition, Ab partially recovered the damage in fetal morphology and bone parameters. Our findings indicated that the co administration of Ab improved the toxicity caused by Pb, and the mushroom could be used as a natural alternative as a protective/chelator agent.

Amelioration of silymarin against cadmium-induced toxicity in pregnant rats and their fetuses.

Silymarin is an antioxidant without side effects even at relatively high physiological dosage. Therefore, it is safely used as a herbal medicine for treating different diseases. The purpose of this study was to examine the toxicity of cadmium (Cd) in pregnant rats and their fetuses and the ameliorative effects of silymarin (SL) against this toxicity. A total of 24 pregnant rats allocated into four equal groups. Control, silymarin (200 mg/kg), Cd (5 mg/kg), and a combination of Cd and silymarin concurrent from 6 to 20th gestational day. Number of corpora lutea, dams', gravid uteri, placental weights, and likewise fetal body weights and lengths were analyzed as physical parameters. Serum levels of aspartate transaminase, alanine transaminase, creatinine, urea, uric acid, and maternal and fetal liver tissues for malondialdehyde, superoxide dismutase, catalase and glutathione activity were studied. The histology of hepatic and renal tissues for both mothers and fetuses was examined. Data were statistically analyzed by analysis of variance test and Duncan's multiple range test was used to compare group means. The findings evidenced that Cd causes teratogenic abnormalities and histopathological variation in hepatic and renal tissues of both mothers and fetuses. Cd triggers oxidative stress and disrupts liver and kidney function. In Cd + silymarin treated rats exhibited improvement in the pregnancy outcomes, reduced histopathological changes, oxidative stress as well as liver and kidney enzymes. We deduced that using of silymarin during gestation is effective and ameliorate the toxic maternal complications caused by cadmium.