Fragments Of Placenta Research Articles

Molecular Detection of Toxoplasma gondii in goats Placenta in Northeastern Algeria

Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis of crucial medical and veterinary importance. It is diagnosed mainly by serological methods, which are insufficiently sensitive. It is therefore necessary to rely on the direct detection of the parasite. The present study aimed at the direct detection of T. gondii DNA in placenta fragments by PCR targeting the B1 gene of the parasite. In addition, we identified possible risk factors for infection. The study was carried out on 25 goat farms between 2019 and 2020 in four regions of the Tebessa province in northeastern Algeria and involved 503 goats that had aborted. T. gondii DNA was detected in 30.41% of goat placenta samples (CI: [27.12-41.53]). The on-farm molecular prevalence was 60% (CI: [53.32-72.53]). The molecular prevalence of toxoplasmosis was higher for primiparous females (53.2%) than it was for multiparous ones (15.2%) (p = 0.0001). Likewise, the level of contamination was high in farms with a sedentary management system (36.4%); this system tended to increase the prevalence of toxoplasmosis (P = 0.008). In addition, it appears that the stage of gestation, history of abortion, and season had a significant effect (p < 0.05) on the prevalence of the disease. To conclude, this study revealed that goats included in this experiment were heavily infected with T. gondii, which represents a major risk for consumers in the Tebessa region. Further research is needed to improve our knowledge of the different genotypes of T. gondii infecting populations of small ruminants.

Glutathione S-transferase (GST) and cortisol levels vs. microbiology of the digestive system of sheep during lambing

BackgroundDuring parturition, animals exhibit variation in hormone levels, homeostasis disturbance and dysfunction of the immune system as a result of stress. Glutathione S-transferase (GST) is responsible for the occurrence of oxidative stress in the cells. Cortisol is known as the stress hormone, but it is also involved in the metabolism of proteins, carbohydrates and metabolism processes led by adipose tissue. The aim of the this study was to determine how the levels of GST and cortisol change depending on the parity. Additionally, the influence of lambing on the microbiological composition of the digestive system and placenta in Olkuska sheep was investigated.MethodsEighteen ewes were selected for the experiment - primiparas (n = 9) and multiparas (n = 9), they were kept in the same environmental conditions, had the same diet and did not show any disease symptoms. Fecal samples were collected individually from each ewe (n = 18) and then bacterial DNA isolation was made, then qPCR analysis for Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria phyla and Lactobacillaceae family bacteria levels was performed. These samples were also used to analyze cortisol levels by ELISA test. In addition, placenta fragments were collected during delivery, and then the GST level from the tissue was tested.ResultsThe analysis of the results showed a higher level of cortisol in primiparous sheep than in multiparous ones, as in the case of glutathione transferase. There were differences between both studied groups in the microbiological composition of the digestive system. In primiparous sheep, the levels of the tested microorganisms were significantly lower than in multiparous ones. A similar relationship occurred in the study of the placental microbiome.ConclusionThe results show that sheep microbiome, cortisol and GST levels are different in primiparas and multiparas. The study conducted may constitute an introduction to further analyzes that would help positively affect the welfare and homeostasis of the female organism.

Associations between SNPS in the genes encoding urokinase system proteins and the risk of placental insufficiency

Placental insufficiency (PI) and its complications are multifactorial conditions that cause perinatal morbidity and mortality. Since the urokinase system is involved in placentation, it should have a role in PI pathogenesis. The aim of this work was to study the associations between single nucleotide polymorphisms (SNPs) of genes coding for protein components of the urokinase system and PI, as well as investigate their effect on the expression of these proteins in the placenta and placental structure. We examined 114 women with uncomplicated pregnancy and delivery, 48 female patients with pre-eclampsia and/or intrauterine growth restriction (IUGR), and 95 newborns, (pre-eclampsia and/or IUGR: n = 60; uncomplicated pregnancy and delivery: n = 35). Maternal and fetal DNAs were genotyped using real-time PCR. Placenta fragments were subjected to morphometry and immunohistochemistry. We discovered the associations between PI and the maternal C allele of rs4065 (PI group: СС-СТ 64.1%, TT 35.9%; controls: СС-СТ 25.6%, TT 74.49%; OR (95%CI): 6.83 (2.63–17.79)), the maternal A allele of rs2302524 (GG-GA 20.5%, AA 79.5% vs. GG-GA 48.1%, AA 51.9%, OR (95%CI): 0.27 (0.1–0.71)), the fetal C allele of rs4065 (СС-СТ 76.4 %, TT 23.6% vs. СС-СТ 69.6%, TT 30.4%, OR (95%CI): 1.37 (0.45–4.17)), and the fetal C allele of rs344781 (TT-TC 69.1%, СС 30.9% vs. TT-TC 95.7%, СС 4.3%, OR (95% CI): 5.02 (1.07–23.6)). The multivariate analysis confirmed the significance of the fetal rs4065 genotype. In patients with PI, uPA expression was lower (ME (95%CI): 116.45 (100.5; 128.74) vs. 126.09 (113.76; 139.19); р < 0.05). No associations were established between SNPs and protein expression. The degree of vascularization depended on the maternal rs4065 genotype (the stroma-to-vessel ratio for the CC genotype was 0.17 (0.15; 0.19); for the CT genotype, 0.18 (0.15; 0.21) and for the TT genotype, 0.23 (0.2; 0.27); p < 0.05). We conclude that high placental uPA and the presence of the fetal TT rs4065 genotype are protective against the risk of PI.

Cerebral injuries associated with Zika virus in utero exposure in children without birth defects in French Guiana: Case report.

Rationale:A major epidemic of Zika virus (ZIKV) infection occurred in French Guiana and West Indies. French national epidemiological surveillance estimated that 1650 pregnant women contracted the ZIKV during epidemic period from January 2016 to October 2016 in French Guiana.Patient concerns:ZIKV infection during pregnancy is a cause of microcephaly and birth defects.Diagnoses:In this report, we describe 2 children with proven in utero ZIKV exposure. Their mothers were both symptomatic and ZIKV infection occurred early in pregnancy. Ultrasonography monitoring in utero did not show any abnormality for both patient. They were born at full-term, healthy, without any birth defects and no sign of congenital ZIKV infection.Interventions:ZIKV was neither found on placenta fragments nor children blood and urine at birth. Their neurodevelopment outcomes in early-life fitted the expectations. As recommended in national guidelines, we performed cerebral MRIs at 2 months old, showing severe brain abnormalities, especially of white matter areas. After a large screening, we did not find any differential diagnosis for their brain lesions.Outcomes:We concluded it was due to their in utero ZIKV exposure.Lessons:In this report, pathogenicity of ZIKV may involve mother's immunological response or metabolic disorder during the infection.

Імплантація кріоконсервованих фрагментів плаценти людини відновлює прооксидантно-антиоксидантний баланс у експериментальних тварин пізнього онтогенезу

The studies have shown that animals of late ontogeny had the misbalance in prooxidative-antioxidative system of blood serum, liver and gonads with shift to the proxidant activity. Nevertheless, the implantation of cryopreserved human placenta fragments to experimental animals increased the activity of endogenous antioxidant system and contributed to the correction of pathological changes associated with aging. Analysis of the findings indicated that the restoration processes occurred both at cell and organism levels. The use of cryopreserved placental fragments in pre-senile age led to a gradual diverse correction of senile involutions, and an increased activity of endogenous antioxidant system during their sanogenesis substantiated the perspective of preventive and therapeutic protocols to treat geriatric pathologies. Probl Cryobiol Cryomed 2017; 27(1): 61–70

Cytochrome P450scc-dependent metabolism of 7-dehydrocholesterol in placenta and epidermal keratinocytes

The discovery that 7-dehydrocholesterol (7DHC) is an excellent substrate for cytochrome P450scc (CYP11A1) opens up new possibilities in biochemistry. To elucidate its biological significance we tested ex vivo P450scc-dependent metabolism of 7DHC by tissues expressing high and low levels of P450scc activity, placenta and epidermal keratinocytes, respectively. Incubation of human placenta fragments with 7DHC led to its conversion to 7-dehydropregnenolone (7DHP), which was inhibited by dl-aminoglutethimide, and stimulated by forskolin. Final proof for P450scc involvement was provided in isolated placental mitochondria where production of 7DHP was almost completely inhibited by 22R-hydroxycholesterol. 7DHC was metabolized by placental mitochondria at a faster rate than exogenous cholesterol, under both limiting and saturating conditions of substrate transport, consistent with higher catalytic efficiency (kcat/Km) with 7DHC as substrate than with cholesterol. Ex vivo experiments showed five 5,7-dienal intermediates with MS spectra of dihydroxy and mono-hydroxy-7DHC and retention time corresponding to 20,22(OH)27DHC and 22(OH)7DHC. The chemical structure of 20,22(OH)27DHC was defined by NMR. 7DHP was further metabolized by either placental fragments or placental microsomes to 7-dehydroprogesterone as defined by UV, MS and NMR, and to an additional product with a 5,7-dienal structure and MS corresponding to hydroxy-7DHP. Furthermore, epidermal keratinocytes transformed either exogenous or endogenous 7DHC to 7DHP. 7DHP inhibited keratinocytes proliferation, while the product of its pholytic transformation, pregcalciferol, lost this capability. In conclusion, tissues expressing P450scc can metabolize 7DHC to biologically active 7DHP with 22(OH)7DHC and 20,22(OH)27DHC serving as intermediates, and with further metabolism to 7-dehydroprogesterone and (OH)7DHP.

The structural heterogeneity of chorial villi phenotype determined by angiogenesis. Implications in legal pathology

Micro anatomic phenotype of chorial villi can be achieved only by means of a rigurous evaluation of its structural elements: trophoblast, vascular and mesenchyme. The authors proposed themselves to study the reciprocal relations between syncytiotrophoblast, fetal sinusoid capillaries and argentic collagen fiber fascicles inside chorial villi depending on angionesis process. The research was carried out on human biologic material using placenta fragments during 28-37 weeks of gestation. The authors consider that the collagen IV stereo distribution inside the vascular pedicle of the terminal villi, contributes to the stability, biodynamic and biokinematics of villi phenotype that is determined by branching or non branching angiogenesis. The personal results have a great value for stating the variability limits of terminal villi phenotype in ortology as well as in general or forensic pathology. Key W ords: terminal villi, angiogenesis, collagen IV, villi phenotype

Dopamine effects the in vitro basal secretion of rat placenta opioids in an opioid and dopamine receptor type-specific manner

Opioid peptides and their receptors are present in the placenta of many species. Dopamine plays an important role in the regulation of opioid release in the nervous system and it may play a similar role in placenta since dopamine receptors are also present in this tissue. The aim of the present work was to examine the effect of dopamine on the basal release of rat placental opioids. The effect of several dopamine receptor agonists and antagonists was tested on the release of immunoreactive β-endorphin and immunoreactive dynorphin from perfused rat placenta fragments. We found that dopamine and apomorphine stimulated the secretion of immunoreactive β-endorphin in a dose-dependent manner. The selective D 1 dopamine receptor agonist (±)-1-phenyl-2,3,4,5-tetrahydro-(1 H)-3-benzazepine-7,8-diol hydrochloride or SKF-38393 reproduced the effect of dopamine while the selective D 1 dopamine receptor antagonist R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl 1,2,3,4,5-tetrahydro-1 H-benzazepine hydrochloride or SCH-23390, prevented the dopamine- and SKF-38393-induced increase of immunoreactive β-endorphin secretion. The selective and potent D 2 dopamine receptor agonist (±)-2-( N-phenylethyl- N-propyl)amino-5-hydroxytetralin hydrochloride or PPHT had no effect on immunoreactive β-endorphin. Finally, none of the agonists tested had any effect on the in vitro secretion of placental immunoreactive dynorphin. Our results suggest that dopamine affects the basal release of placental opioids in an opioid and dopamine receptor-specific manner, its effect being different from the effect it exerts on β-endorphin in the rat neurointermediate pituitary lobe.

Anion transport by human placenta: a study of chloride and sulphate efflux from isolated placental tissue fragments.

The efflux of radiolabelled sulphate and chloride from fragments prepared from normal human term placentae has been examined so that a comparison between the transport properties of whole tissue and isolated placental membranes may be made. Sulphate efflux was found via a temperature- and DIDS sensitive mechanism. External chloride and sulphate were able to trans-accelerate the egress of labelled sulphate from tissue fragments via a DIDS-inhibitable route. These results are consistent with sulphate transport being mediated via an anion-exchange process. Chloride efflux from fragments of placenta was via a system which could be trans-stimulated by external Cl-: this moiety of transport was inhibited by DIDS. Several differences between the characteristics of placental tissue anion transport and isolated membrane vesicles were noted and are discussed.

Effect of Parathyroid Hormone on PO4 Transport through the Human Placenta Microvilli

The transport of phosphate (PO4) through the placenta is a secondary active phenomenon whose control mechanisms are unknown. In this study, we investigated whether PTH, the main hormone regulating PO4 transport in the kidney and gut, has a similar role in the placenta. Incubation of normal term human placenta fragments for 1 min with PTH increased the cAMP content of the tissue by 285%. A dose-response curve of the effect of the hormone showed that the cAMP accumulation reached a maximal level with 3.5 x 10(-8) M PTH. Incubation of the placenta fragments with 10(-4) M di-butyryl cAMP resulted in a significant decrease in the PO4 uptake by the brush border membranes prepared from these fragments. Increasing concentrations of di-butyryl cAMP from 0 to 10(-3) M significantly decreased the PO4 uptake from 0.29 +/- 0.02 to 0.22 +/- 0.01 pmol/micrograms/20 s. Similarly, incubation of the placental tissue with PTH resulted in a comparable decrease in the PO4 uptake by the corresponding brush border membrane vesicles. In contrast, direct incubation of brush border membranes with the hormone did not influence PO4 uptake. It is concluded that PTH probably regulates the PO4 transport through the placenta syncytio-trophoblast cell through cAMP mediation. Because adenylate cyclase is located in the basal plasma membrane, it is likely that only the fetal hormone is implicated in this process.

Extracellular Volume of the Human Placenta in vitro

The extracellular volume of fresh and perfused human placenta has been measured in two sets of experiments. In the first set fragments of placenta which had been dually perfused for either 45 or 110 min as well as fragments from unperfused areas of the same placenta were incubated for 4 h in Earle's bicarbonate buffer containing 51Cr-EDTA as an extracellular marker. The 51Cr-EDTA space, as a proportion of wet weight, was 76.5 +/- (SE) 1.4 (n = 11) and 79.4 +/- 1.8% (n = 6) for the short and long perfusions, respectively; these did not significantly differ from each other or from the values for the equivalent unperfused placenta: 75.2 +/- 1.0 (n = 11) and 77.7 +/- 0.9% (n = 6). In the second set of experiments 51Cr-EDTA and 14C-inulin were perfused through the maternal and fetal circulations of the placental lobules for 1, 2, or 4 h, and simultaneously unperfused fragments from the same placenta were incubated with the markers for 1, 2, or 4 h. The 51Cr-EDTA space was found to be significantly higher than the 14C-inulin space in both perfusion and incubation determinations. Irrespective of the marker, the incubation spaces were unaltered with time and were significantly higher than the perfusion spaces at all three time points. However, the difference decreased with the length of perfusion, so that by 4 h it was less than 10%. (At 4 h incubation the 51Cr-EDTA space was 76.2 +/- 0.9% and the perfusion space 67.4 +/- 1.1%, n = 6; the 14C-inulin incubation space was 60.9 +/- 2.2% and the perfusion space 52.5 +/- 2.6%, n = 5). As there was no evidence of increasing placental permeability with time, as judged by clearance of creatinine, it was concluded that the placental extracellular volume is large irrespective of the method of measurement.

Release of Mammotrophic Activity by the Rat Placenta

Release of mammotrophic activity into the culture medium by placenta fragments in vitro could be demonstrated with the aid of an organ culture of mammary explants. Maintenance of the placenta fragments was optimal with less than 8 fragments per ml medium. Release of activity was already high after one hour of culture, but continued for at least four days if the medium was favourable. To obtain this latter effect the presence of salts and glucose in the medium was sufficient, amino acids, serum and insulin being of no or minor importance. The activity observed can be explained partly by a slow release of hormone(s) already present in the fragments. Activity was released by freezing and thawing the placenta fragments. If placenta fragments were kept at + 4°C or + 20°C release of activity continued for several days. Moreover, the release at + 37°C was not inhibited by cycloheximide. On the other hand intact fragments released more activity than frozen fragments, even after one day of culture, while the release continued at a high level for several days in intact fragments and stopped after one day in frozen fragments. The placenta fragments had an active metabolism with glucose consumption and lactate production. When virgin rat serum was incubated with placenta fragments in vitro, its mammotrophic activity became equal to that of pregnant rat serum. Pregnant rat serum seemed to inhibit the release of mammotrophic activity from placenta fragments in some experiments, but the assay method does not permit a definite conclusion. The medium activated by incubation with placenta fragments showed synergism with cortisol as regards secretion, and with progesterone as regards proliferation. An effect of cortisol, progesterone, oestradiol, oestrone or oestriol on the release of mammotrophic activity from the placenta fragments could not be demonstrated. Rat placental giant cells seem to be involved in the production of rat chorionic mammotrophin (rCM). Activity appeared in the maternal serum at day 8-9 of pregnancy when mainly the trophoblastic giant cells adjacent to the foetal membranes are in contact with the maternal circulation. Those placenta fragments which on organ culture released activity into the culture medium contained giant cells. Undifferentiated giant cells and/or basal zone cells also may be capable of producing rCM. The giant cells could be maintained viable in vitro. They incorporated thymidine, uridine and leucine. The giant cells did not stain with acidophilic staining for pituitary hormones. Electron microscopy suggested that if the giant cells do in fact produce rCM, the Golgi apparatus may not be involved in its secretion. Secretion granules could not be demonstrated in the giant cells. Sephadex G-100 gel chromatography fractions were obtained from pregnant rat serum, placenta extract and placenta culture medium. The fractions were tested for the presence of mammotrophic activity. In 13 and 19 days pregnant rat serum the fraction with molecular weight of approximately 35 000 showed high activity. In placenta extracts obtained from 13 and 19 days pregnant rats activity was present in the fraction with molecular weight of approximately 18 000, but other fractions with lower and higher molecular weight also possessed high mammotrophic activity. Media collected from a placenta culture similarly contained multiple fractions with activity.

The production of Mullerian inhibiting substance by the fetal, neonatal and adult rat.

The intensity and duration of Mullerian Inhibiting Substance (MIS) secretion by the fetal newborn and adult rat testes and the effect of the placenta and pituitary on the secretory pattern was studied in vivo and in vitro. A graded organ culture assay was used to determine the secretory pattern. Production of MIS was evident during the last trimester of pregnancy. MIS production continued during the 1st 3 weeks of neonatal life and then disappeared around the time of weaning. Hypophysectomy performed at 20 days of age failed to restore MIS activity in adult animals. Fragments of placenta in testis culture did not influence MIS activity in perinatal testes. MIS exhibited a regressive effect on Mullerian ducts placed at a distance from a testicular source.

Studies on accumulation and metabolic fate of [ N-Me 3H]choline in human term placenta fragments

Fragments from human term placenta accumulated [ N-Me 3H]choline against a concentration gradient. Uptake was linearly related to incubation time and temperature. Analysis of the kinetics of choline accumulation revealed the concurrent existence of a diffusional component occuring both at low temperature as well as high (50 mM) choline concentration and a carrier-mediated transport which had characteristics predicted by the Michaelis-Menten equation showing a K m = 3.46 × 10 −4 M and a V max of 75 nmoles/ml intracellular water × min −1. Net concentration ratios, corrected for diffusion and extracellular water, were larger than 1.0 within 5 min and about 4.0 after 15 min. Hemicholinium-3 was a competitive inhibitor of choline uptake with a K i of 0.45 mM. [ 3H]Choline accumulation was decreased by conditions known to lower intracellular ATP levels. Thus, 2,4-dimitrophenol (1 mM), sodium cyanide (5 mM) and anaerobic incubation reduced [ 3H]choline accumulation 36, 54 and 33 per cent respectively. Ouabain (0.1 mM) also decreased the concentration ratios by 50 per cent. Modification of the ionic environment led to an increase of 36 per cent in the amount of tritium in intracellular water when Na + was reduced to one half of the usual 145 mM or 150 per cent when it was completely omitted and replaced by an osmotically equivalent amount of sucrose. Li + was without effect, while high K + (> 25mM), Rb + and Cs + (145 mM) depressed [ 3H]choline accumulation. The metabolic fate of [ 3H]choline was studied. Following a 5-min incubation with 5 μM [ 3H]choline 95 per cent of the radioactivity was acid-soluble and 5 per cent remained in the acid-insoluble fraction. After 30 min the distribution was 88 and 12 per cent, respectively. Paper high voltage electrophoretic analysis of the acid-soluble material showed that after 5 min 55 per cent of the 3H had a mobility equal to authentic choline, 35 per cent equal to acetylcholine, 6 per cent to phosphorylcholine and 1 per cent to betaine. After 20 min it was 25 per cent in choline, 60 per cent in acetylcholine, 10 per cent in phosphorylcholine and 2 per cent in betaine. A chloroform-methanol extract from the acid-insoluble residue revealed a linear increase of 3H-content suggesting incorporation of [ 3H]choline into phospholipids.

Uptake of acetylcholine by human placenta fragments and slices from guinea pig and rat placenta

Fragments of human term placenta took up [ 3H]acetylcholine (ACh) into the intracellular water fraction against a concentration gradient when incubated in a medium which contained 10 μM paraoxon as an inhibitor of cholinesterases. Intracellular concentrations rose to levels 3- to 4-fold over the concentration in the medium in a time- and temperature-dependent manner. The ACh uptake was saturable and followed the Michaelis-Menten equation. It required metabolic energy and was markedly reduced by drugs and conditions which decreased cell levels of ATP. Concentrative uptake was inhibited by alterations in the ionic environment as introduced by high K +, Li +, Rb + and Cs +. Slices from guinea pig placenta reached intracellular concentrations which were only equal to the concentration in the incubation medium, suggesting equilibration by diffusion, while rat placenta slices concentrated ACh more slowly than human placenta and reached ratios of 2.0. These species variations further confirm known differences of the cholinergic system in placenta and stress the need for caution in extrapolating findings from the placenta of one species to another.

Secretion of human chorionic gonadotrophin by the early placenta in Vitro.

The secretion in vitro of HCG and proteins was studied in fragments of placenta from women in the first trimester of pregnancy by a pulse-chase system. A 10-min pulse with [3H]leucine was used. It was concluded that the approximate half-time of release of HCG was 150 min. Proteins precipitable with trichloroacetic acid had a bi-exponential pattern, the half-times of release being 100 and 270 min. These rates of release indicate that the HCG produced by the early placenta was rapidly passed into the circulation rather than stored.

Vacuum aspiration in the delivery room.

Abstract. A vacuum aspirator was used to remove retained fragments of placenta, membranes and hyperplastic decidua after delivery. No post‐partum endometritis occurred in the group of 60 patients so treated, whereas five infections were found in a control group of 50 patients. The difference is almost significant. Moreover, favourable experience has led to the use of the vacuum aspirator in the treatment of atonic uterine haemorrhage. No complications have been observed, and the risk seems to be less than that in conventional curettage, particularly as anaesthesia is unnecessary.