PlGF Levels Research Articles

Abstract 25 Cord Blood Mononuclear Cells and Cord Blood Plasma Alter Human Cardiac Endothelial Cell Angiogenic Sprouting and Growth In Vitro

Abstract Introduction The mononuclear cell fraction of human placental cord blood (CBMNC) comprises immune cells, hematopoietic, mesenchymal and other stem or progenitor cells. CBMNCs are proposed to afford therapeutic benefit based on their paracrine release of multiple cytokines, growth factors and other proteins that stimulate signalling for wound healing, including attenuation of inflammation and fibrosis. Whether CBMNCs augment angiogenic sprouting and tube formation by endothelial cells in human ventricular myocardium is not well characterised. Objectives The aim of this study was to investigate the in vitro influence of CBMNCs on endothelial cell angiogenic-like growth. Methods A tube formation assay for human donor primary ventricular myocardium microvascular endothelial cells (HMVECs), (3 female donors, age 48-50 yrs, 3 replicates per treatment group) was established using Matrigel-coated 96-well plates (62,500-78,125 cells/cm2). Cells from passage 5-8 were used for all experiments.. Angiogenic processes, as characterized by endothelial sprouting followed by tube extension into honeycomb-like mesh structures, were imaged over 24 hours. HMVECs were treated with CBMNCs at a concentration of 1:10 (MNC:HMVEC) or cord blood plasma (1μg/μL) at 0h, 3h, or 6h after the HMVECs were seeded. The cells were visualized by phase contrast microscope and images (4x magnification) were analysed using the Angiogenesis Analyzer plugin in FIJI ImageJ. Size exclusion chromatography and multiplex ELISAs were performed to determine the presence of analytes in media collected from each well. Results CBMNC-treated HMVECs exhibited greater average total elongated branching length and more tube-like structured meshes, compared to untreated controls. MMP-1, MMP-2, MMP-9, MMP-10, TIMP-1, TIMP-2, angiopoietin-2, EGF, endoglin, endothelin-1, FGF-2, follistatin, HB-EGF, IL-8, PLGF, VEGF-A, and VEGF-C were detected in extracellular media during in vitro tube formation. With CBMNC treatment, only MMP-9 was increased by 475.94%, whereas angiopoietin-2, follistatin, and PLGF levels were decreased by 27.90%, 15.90%, and 26.18% respectively. In contrast, CB plasma-treated HMVECs did not form tube sprouts, instead an intense endothelial cell proliferation was observed which was associated with increased MMP-2, MMP-9, endothelin-1 and IL-8 levels with decreased angiopoietin-2, HB-EGF, PLGF, and endoglin levels. Discussion CBMNC treatment enhanced pro-angiogenic processes in HMVECs in vitro through increased branching and network formation, whereas CB plasma evoked cell proliferation.

Abstract P423: INCREASED LEVELS OF URINARY ANGIOGENIC FACTORS AND MARINOBUFAGENIN IN PREECLAMPSIA PATEINT

Background: Preeclampsia (preE) is a syndrome which occurs in 3-10% of pregnancies and is a leading cause of maternal and fetal morbidity and mortality. The precise etiology (etiologies) of this syndrome remain(s) unknown. There is no single biomarker for detection of the syndrome. Objective: To measure angiogenic, antiangiogenic factors, and MBG in urine of pregnant patients with and without preE. Methods: A prospective cohort of patients with normal pregnancies and of those with preE were recruited. sFlt-1, soluble endoglin (sEng), PlGF, TGF-β-1, VEGF, and MBG were assayed using ELISA and results corrected to units per mg creatinine. Patient characteristics were compared using Chi-square tests for proportions, Student’s t test for parametric data, and Mann-Whitney U test for nonparametric data. P < 0.05 was taken as significant. Variation associated with gestational age was examined using correlations and when appropriate linear regression equations were obtained. Receiver operator curves (ROC) were examined to establish criteria levels for those factors that varied with preeclampsia. Results: In total, 40 patients with normal pregnancies were sampled between 22 and 39 weeks gestation and 30 patients with preE were sampled between 28 and 39 weeks gestation. As expected, groups differed (p < 0.02) for blood pressure, gestational age at delivery, and age. They did not differ for weight (p = 0.28), height (p = 0.38), and creatinine levels (p = 0.29). Groups differed in sEng (p = 0.016), PlGF (p = 0.018), sFlt-1 (p = 0.00066), and MBG (p < 0.0001). The PlGF levels were less in preE while sEng, sFlt-1 and MBG levels were increased. TGF-β-1 and VEGF did not differ between groups (p > 0.14). Only urinary MBG was found to vary with gestational age in preE (p = 0.0009). MBG, sFlt-1, PlGF, and sFlt-1/PlGF ratio had significant ROC area under the curve measures (p < 0.02) with criteria for preeclampsia that provided sensitivities between 65 and 88% and specificities between 74 and 96%. Conclusions: Urinary levels of angiogenic and antiangiogenic factors and MBG as measured in urine demonstrated differences in patients with clinical diagnosis of preE. Urine specimen procurement and assay are less invasive and potentially more acceptable for assessment of preE.

EFFECT OF MELATONIN ON THE LEVEL OF CYTOKINES IN PREECLAMPSIA

Introduction. The literature reports a decrease in melatonin levels in preeclampsia, when the degree of decrease correlates with the severity of the process, as well as a decrease in the expression of melatonin receptors in the placental tissue in case of fetal growth restriction.Regarding changes in the cytokine profile in PE: there is evidence that the levels of proinflammatory cytokines, namely TNF-α and IL-6, increase in preeclampsia, while the concentrations of anti-inflammatory cytokines, namely IL-4 and IL-10, decrease. Melatonin, on the contrary, reduces the secretion of proinflammatory cytokines, in particular TNF-α, and increases the production of anti-inflammatory cytokines, namely IL-10. Therefore, normalization of melatonin levels may be a promising direction in the treatment of PE in pregnant women.Objective of the study. Assess the pathogenetic mechanisms of complications arising from a decrease in melatonin, and, in particular, find out more about the changes in melatonin, IL-6 and IL-10, determine the correlation between them.Material and research methodsThe study conducted at the Department of Pregnancy Pathology of the Chernivtsi Regional Perinatal Center, involved 32 women whose pregnancy was complicated by preeclampsia. The control group consisted of 33 women with uncomplicated pregnancies observed in the antenatal clinic of the Chernivtsi Regional Perinatal Center.The patient's informed consent form and the patient's examination card were approved by the Biomedical Ethics Committee of Bukovinian State Medical University (BSMU) of the Ministry of Health of Ukraine (Chernivtsi). The work was guided by the general provisions of the Declaration of Helsinki "Recommendations for Physicians for Biomedical Research Involving Human Subjects" (1964), the World Medical Association on Ethical Principles for Scientific Medical Research Involving Human Subjects (1964-2000), considering the requirements of Directive 2001/20/EC of the European Parliament and of the Council, ICH GCP, the Council of Europe Convention on Human Rights and Biomedicine (04.04.1997), Order of the Ministry of Health of Ukraine No. 690 of 23.09.2009.Statistical data were calculated using MedCalc software developed by MedCalc Software (Ostend, Belgium). The results were evaluated using the Mann-Whitney U-test for small groups. The P value of <0.05 was considered valid. Correlation and regression statistical evaluations were also performed using the aforementioned software.R&D «Preservation and restoration of reproductive health of women and girls in obstetric and gynecological pathology». State registration number: 0121U110020. Terms of execution: 01.2021–12.2025 yy.Research results. The level of melatonin in venous blood taken from women with diagnosed PE was significantly lower (p=0.029), as well as levels of placental growth factor PlGF (p<0.0001), compared to healthy women. In our study we also found that the concentrations of both proinflammatory IL-6 and anti-inflammatory IL-10 were elevated in women with PE compared to women with uncomplicated pregnancies. We performed a prognostic assessment of the impact of the biochemical parameters we studied on the outcome of pregnancies in the examined patients. Melatonin, if present in the blood of a pregnant woman in a concentration higher than the threshold values we established (6.71 pg/ml), reduces the chances of clinical manifestations of severe preeclampsia with a high degree of reliability (p=0.00173). We assume that placental dysfunction in particular leads to impaired synthesis of the antioxidant melatonin, which causes disorders in the tissues of the mother, placenta and fetus and, as a result, provokes clinical manifestations of preeclampsia. The condition of the placenta also worsens the subclinical inflammatory process, as evidenced by an increase in IL-6 levels in patients with diagnosed preeclampsia; an increase in IL-10 in the study group, in our opinion, is a kind of compensatory reaction that prevents preterm labour. In women with pregnancies complicated by preeclampsia, melatonin is a prognostic criterion for delivery before 38 weeks of gestation, and this delivery is mainly associated with the progression of preeclampsia to its severe stage. In the group of patients with uncomplicated pregnancy, proinflammatory interleukin-6 is predictive of spontaneous onset of labour before 38 weeks of gestation. Placental growth factor PlGF was a predictor of the birth of children with a body weight of less than 3000 g in both study groups.Conclusions. Concentrations of melatonin and of placental growth factor in venous blood in women whose pregnancy was complicated by preeclampsia in the third trimester of pregnancy are significantly lower compared to uncomplicated pregnancies. On the other hand, the levels of proinflammatory IL-6 and anti-inflammatory IL-10 were significantly increased in the group of patients with diagnosed preeclampsia compared with normal pregnancy.

The genetics of autism and steroid-related traits in prenatal and postnatal life.

Autism likelihood is a largely heritable trait. Autism prevalence has a skewed sex ratio, with males being diagnosed more often than females. Steroid hormones play a mediating role in this, as indicated by studies of both prenatal biology and postnatal medical conditions in autistic men and women. It is currently unclear if the genetics of steroid regulation or production interact with the genetic liability for autism. To address this, two studies were conducted using publicly available datasets, which focused respectively on rare genetic variants linked to autism and neurodevelopmental conditions (study 1) and common genetic variants (study 2) for autism. In Study 1 an enrichment analysis was conducted, between autism-related genes (SFARI database) and genes that are differentially expressed (FDR<0.1) between male and female placentas, in 1st trimester chorionic villi samples of viable pregnancies (n=39). In Study 2 summary statistics of genome wide association studies (GWAS) were used to investigate the genetic correlation between autism and bioactive testosterone, estradiol and postnatal PlGF levels, as well as steroid-related conditions such as polycystic ovaries syndrome (PCOS), age of menarche, and androgenic alopecia. Genetic correlation was calculated based on LD Score regression and results were corrected for multiple testing with FDR. In Study 1, there was significant enrichment of X-linked autism genes in male-biased placental genes, independently of gene length (n=5 genes, p<0.001). In Study 2, common genetic variance associated with autism did not correlate to the genetics for the postnatal levels of testosterone, estradiol or PlGF, but was associated with the genotypes associated with early age of menarche in females (b=-0.109, FDR-q=0.004) and protection from androgenic alopecia for males (b=-0.135, FDR-q=0.007). The rare genetic variants associated with autism appear to interact with placental sex differences, while the common genetic variants associated with autism appear to be involved in the regulation of steroid-related traits. These lines of evidence indicate that the likelihood for autism is partly linked to factors mediating physiological sex differences throughout development.

Associations Between CSF Markers of Inflammation, White Matter Lesions, and Cognitive Decline in Individuals Without Dementia.

Small vessel disease (SVD) and neuroinflammation both occur in Alzheimer disease (AD) and other neurodegenerative diseases. It is unclear whether these processes are related or independent mechanisms in AD, especially in the early stages of disease. We therefore investigated the association between white matter lesions (WML; the most common manifestation of SVD) and CSF biomarkers of neuroinflammation and their effects on cognition in a population without dementia. Individuals without dementia from the Swedish BioFINDER study were included. The CSF was analyzed for proinflammatory markers (interleukin [IL]-6 and IL-8), cytokines (IL-7, IL-15, and IL-16), chemokines (interferon γ-induced protein 10, monocyte chemoattractant protein 1), markers of vascular injury (soluble intercellular adhesion molecule 1, soluble vascular adhesion molecule 1), and markers of angiogenesis (placental growth factor [PlGF], soluble fms-related tyrosine kinase 1 [sFlt-1], vascular endothelial growth factors [VEGF-A and VEFG-D]), and amyloid β (Aβ)42 Aβ40, and p-tau217. WML volumes were determined at baseline and longitudinally over 6 years. Cognition was measured at baseline and follow-up over 8 years. Linear regression models were used to test associations. A total of 495 cognitively unimpaired (CU) elderly individuals and 247 patients with mild cognitive impairment (MCI) were included. There was significant worsening in cognition over time, measured by Mini-Mental State Examination, Clinical Dementia Rating, and modified preclinical Alzheimer composite score in CU individuals and patients with MCI, with more rapid worsening in MCI for all cognitive tests. At baseline, higher levels of PlGF (β = 0.156, p < 0.001), lower levels of sFlt-1 (β = -0.086, p = 0.003), and higher levels of IL-8 (β = 0.07, p = 0.030) were associated with more WML in CU individuals. In those with MCI, higher levels of PlGF (β = 0.172, p = 0.001), IL-16 (β = 0.125, p = 0.001), IL-8 (β = 0.096, p = 0.013), IL-6 (β = 0.088, p = 0.023), VEGF-A (β = 0.068, p = 0.028), and VEGF-D (β = 0.082, p = 0.028) were associated with more WML. PlGF was the only biomarker that was associated with WML independent of Aβ status and cognitive impairment. Longitudinal analyses of cognition showed independent effects of CSF inflammatory markers and WML on longitudinal cognition, especially in people without cognitive impairment at baseline. Most neuroinflammatory CSF biomarkers were associated with WML in individuals without dementia. Our findings especially highlight a role for PlGF, which was associated with WML independent of Aβ status and cognitive impairment.

Vascular endothelial growth factor-D plasma levels and VEGFD genetic variants are independently associated with outcomes in patients with cardiovascular disease.

The vascular endothelial growth factor (VEGF) family is involved in pathophysiological mechanisms underlying cardiovascular (CV) diseases. The aim of this study was to investigate the associations between circulating VEGF ligands and/or soluble receptors and CV outcome in patients with acute coronary syndrome (ACS) and chronic coronary syndrome (CCS). Levels of VEGF biomarkers, including bFGF, Flt-1, KDR (VEGFR2), PlGF, Tie-2, VEGF-A, VEGF-C, and VEGF-D, were measured in the PLATO ACS cohort (n = 2091, discovery cohort). Subsequently, VEGF-D was also measured in the STABILITY CCS cohort (n = 4015, confirmation cohort) to verify associations with CV outcomes. Associations between plasma VEGF-D and outcomes were analysed by multiple Cox regression models with hazard ratios (HR [95% CI]) comparing the upper vs. the lower quartile of VEGF-D. Genome-wide association study (GWAS) of VEGF-D in PLATO identified SNPs that were used as genetic instruments in Mendelian randomization (MR) meta-analyses vs. clinical endpoints. GWAS and MR were performed in patients with ACS from PLATO (n = 10 013) and FRISC-II (n = 2952), and with CCS from the STABILITY trial (n = 10 786). VEGF-D, KDR, Flt-1, and PlGF showed significant association with CV outcomes. VEGF-D was most strongly associated with CV death (P = 3.73e-05, HR 1.892 [1.419, 2.522]). Genome-wide significant associations with VEGF-D levels were identified at the VEGFD locus on chromosome Xp22. MR analyses of the combined top ranked SNPs (GWAS P-values; rs192812042, P = 5.82e-20; rs234500, P = 1.97e-14) demonstrated a significant effect on CV mortality [P = 0.0257, HR 1.81 (1.07, 3.04) per increase of one unit in log VEGF-D]. This is the first large-scale cohort study to demonstrate that both VEGF-D plasma levels and VEGFD genetic variants are independently associated with CV outcomes in patients with ACS and CCS. Measurements of VEGF-D levels and/or VEGFD genetic variants may provide incremental prognostic information in patients with ACS and CCS.

The correlation between serum levels and placental tissue expression of PLGF and sFLT-1 and the FIGO grading of the placenta accreta spectrum disorders

Objective This study aims to further explore the role of angiogenic vs anti-angiogenic factors in placenta accreta spectrum (PAS). Methods This cohort study included all patients with placenta previa and placenta accreta spectrum (PAS) disorders undergoing surgery at Dr. Soetomo Hospital (Academic Hospital of Universitas Airlangga, Surabaya, Indonesia) from May to September 2021. Venous blood samples for PLGF and sFlt-1 were drawn immediately prior to surgery. Placental tissue samples were taken during surgery. The FIGO grading was diagnosed intraoperatively by an experienced surgeon and confirmed by the pathologist and followed by immunohistochemistry (IHC) staining. The sFlt-1 and PLGF serum were performed by an independent laboratory technician. Results Sixty women were included in this study (20 women with placenta previa; 10 women with FIGO PAS grade 1; 8 women with FIGO PAS grade 2; 22 women with FIGO PAS grade 3). The median with 95% Confidence interval of PLGF serum values in placenta previa, FIGO grade I, grade II, and grade III were 233.68 (0.00–2434.00), 124.39 (10.42–663.68), 236.89 (18.83–418.99) and 237.31 (2.26–3101.00) (p = .736); the median values with 95% CI of serum sFlt-1 levels in placenta previa, FIGO grade I, grade II, and grade III were 2816.50 (418.00–12925.00), 2506.00 (227.50-16104.00), 2494.50 (888.52–20812.00), and 1601.00 (662.16–9574.00) (p = .037). Placental PLGF expression in placenta previa, FIGO grade 1, grade II, and grade III showed median values (with 95% CI) of 4.00 (1.00-9.00), 4.00 (2.00–9.00), 4.00 (4.00–9.00), and 6.00 (2.00–9.00) (p = .001); sFlt-1 expression median values (with 95% CI) were 6.00 (2.00–9.00), 6.00 (2.00–9.00), 4.00 (1.00–9.00), and 4.00 (1.00–9.00) (p = .004). Serum PLGF and sFlt-1 levels did not correlate with placental tissue expression (p = .228; p = .586). Conclusion There are differences in PAS's angiogenic processes ​according to the severity of trophoblast cell invasion. But there is no overall correlation between serum levels and PLGF and sFlt-1 expression in the placenta, suggesting the imbalance between angiogenic and anti-angiogenic are local mechanisms in the placental and the uterine wall.

Clinical development and evaluation of plasma angiogenesis factors from phase II study of FOLFIRI plus ramucirumab with recurrent colorectal cancer refractory to adjuvant chemotherapy with oxaliplatin/fluoropyrimidine (RAINCLOUD): RAINCLOUD-TR.

173 Background: Higher levels of vascular endothelial growth factor-D (VEGF-D) is a potential predictive biomarker for ramucirumab (Ram) efficacy on overall survival and progression-free survival (PFS) in Ram+FOLFIRI vs Placebo+FOLFIRI for bevacizumab (Bev) refractory metastatic colorectal cancer (mCRC). However, it is not clear whether VEGF-D acts as a predictive biomarker in pts who pretreated without Bev. RAINCLOUD showed the clinical benefit of FOLFIRI+ plus Ram for pts with recurrent CRC refractory to adjuvant chemotherapy (median PFS was 6.2 months (M); ESMO-GI 2022). RAINCLOUD-TR was designed as a prospective biomarker study estimating the association of biomarkers with Ram efficacy. Methods: Plasma sample collections were done at time points of pre-/ post-treatments in pts receiving FOLFIRI+Ram. Measurements of 17 factors (including VEGF-A, VEGF-D and PlGF) were performed by multiplex assay with Luminex technology. The high/ low groups were defined bases on the median plasma level at the baseline. Results: From September 2017 to September 2021, 81 plasma samples were collected from 48 pts and 32 pts in both pre/ and post treatment. Level of VEGF-A, VEGF-D, PlGF and TSP-2 were all significantly higher in post vs pre-treatment [Table]. The ORR analyses were as follows; VEGF-A, VEGF-D and PlGF in low vs high were 50.0%/33.3% (p=0.35, HR 1.97), 55.0% /33.3% (p=0.22, HR 2.39), and 60.0%/28.6% (p=0.35, HR 1.95). The PFS analyses were as follows; VEGF-A, VEGF-D, and PlGF in low vs high were 6.1M/6.0M (p=0.815, HR 0.92), 7.6M /5.6M (p=0.095, HR 0.56), and 8.0M/4.2M (p=0.014, HR 0.40). In other factors, low TSP-2 were better ORR (low vs high: 66.7%/20.0% (p=0.004, HR 7.54) and PFS 7.5M/4.3M (p=0.022, HR 0.45). Conclusions: In this exploratory study, PFS was numerically greater for pts with low vs high VEGF-D for Ram in pts with mCRC pretreated without Bev. VEGF-D may not be predictive biomarker, but TSP-2 may be a potential prognostic biomarker, further studies in larger cohorts are needed to confirm our results. Clinical trial information: UMIN000028678 . [Table: see text]

Оцінка показників комплексної стратифікації ризиків розвитку прееклампсії в пацієнток із ретрохоріальними гематомами

Purpose - to assess the prognostic value of a comprehensive study of the influence of indicators of angiogenic/antiangiogenic profile in women with retrochorial hematoma (RCH) in the І trimester, uterine artery (UA) dopplerometry data in stratifying the risks of developing placental dysfunction in these women. Materials and methods. A prospective analysis of the course of pregnancy was carried out in 137 women with a threat of miscarriage aged 20 to 47 years, who made up two comparison groups: the Group I - 60 patients with RCH, the Group II - 77 patients with a threat of abortion without hematoma. Results. The average age of women of the Group I was 31.2±0.6 years, of the Group II - 32.2±0.6 years. The gestational age at enrollment was equal 6.1±0.55 weeks in the Group I and 7.2±0.61 weeks in the Group II. A direct, reliable correlation of weak strength was established in pairs: the presence of the human chorionic gonadotropin (hCG) and the pulsatility index (PI) in UA &gt;95 percentile, hCG and a higher level of hCG in the I and II trimesters of pregnancy. A reduced level of PAPP-A was significantly associated with cases of preeclampsia (PE) in the anamnesis, increased Body Mass Index, a high level of PI in UA, including with PI levels &gt;95 percentile, as well as with a reduced level of free estriol. Significant inverse correlations were established between the level of PLGF and indicators of PE in history, the level of PI in UA and the content of hCG. At the same time, the level of alpha-fetoprotein in the studied patients was directly associated with increased levels of PI in UA and hCG. It was established that the risk of early PE was more inherent for women with the presence of PCH in the І trimester of pregnancy, while the percentage of the development of late PE with and/or without fetal growth retardation was more often higher in women with a threat of termination of pregnancy in the І trimester trimesters without the formation of RHG. Conclusions. The occurrence of RHG at the stage of early placentation increases the risks of developing placental dysfunction and obstetric complications associated with it. PRISCA-1, PLGF, PI of UA, as well as the calculation of the risk of developing PE in the trimester І using the FMF calculator should be used to form a risk group for the development of placenta-associated complications. Indicators of PI of UA &gt;99 percentile in the І trimester of pregnancy in combination with a decrease in PAPP-A ˂0.45 MoM should be considered critical. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patients was obtained for conducting the studies. No conflict of interests was declared by the authors.

Prediction of the development of fetal growth retardation in pregnant women with chronic arterial hypertension

During intrauterine life a fetus with growth restriction does hemodynamic, metabolic and hormonal adjustments to cope with the adverse uterine environment, and these changes can become permanent and irreversible. Despite the progress in the knowledge of delayed fetal development, the biomarkers are able to identify this pathology at an early stage and stratify its severity, both before and after childbirth, are still not determined. The objective: to evaluate the prognostic value of the angiogenesis marker level, hormonal profile and Doppler examination for fetal growth restriction (FGR) in pregnant women with chronic arterial hypertension (CAH).Materials and methods. A prospective examination of 61 pregnant women with CAH 1-2 degrees was conducted, which included an analysis of the clinical and anamnestic characteristics and the study of the pregnancy course in the first trimester (11-12 weeks). To determine the level of risk factors for the FGR development the examined pregnant women were divided into groups: women with CAH with FGR (n=10), and pregnant women with CAH without FGR (n=51).Human chorionic gonadotropin (CHG), progesterone (PG) and estradiol (E) were determined in blood serum. Angiogenesis markers – placental growth factor (P1GF) as a pro-angiogenic factor and soluble fms-like tyrosine kinase (sFlt-1) as an anti-angiogenic factor were studied, the ratio of sFlt-1/PLGF (coefficient K) was calculated. All patients had Doppler examination of vascular blood flow in the left and right uterine arteries (pulsation index – PI, resistance index – RI and systolic-diastolic ratio – SDR). The ultrasound examination was done with the ultrasound machine “Mylab Six System”. Statistical analysis was performed using program Statistica® for Windows 13.0. A single-factor logistic regression analysis and a method of building logistics regression, which was evaluated by means of receive operating characteristic curve (ROC-curve) according to the area under the curve (AUC) were used to determine the prognostic level of markers, the 95 % confidence interval was calculated. Results. In the first trimester of pregnancy in women with CAH the PLGF level was &lt; 8.2 pg/ml (sensitivity – 80.0 %, specificity – 60.78 %), area under the ROC curve (AUC) – 0.697 (95 % CI: 0.566–0.808; p =0.0128), which indicate an increased risk of FGR development. The growth of sFl-1 &gt; 1802.59 ng/ml (sensitivity – 70.0 %, specificity – 64.71 %), area under the ROC-curve (AUC) 0.678 (95 % CI: 0.547–0.792; p=0.05) increase the risk of FGR development. The coefficient K in 11-12 weeks of pregnancy was ≥ 95.36 units (sensitivity – 100.0 %, specificity – 43.14 %), area under ROC-curve (AUC) – 0.735 (95 % CI: 0.607–0.840; p=0.0128), which indicate an increase the risk of FGR development. PG/PlGF ratio is 7.21 (sensitivity – 70.0 %, specificity – 70.59 %) and area under the ROC-curve (AUC) 0.702 (95 % CI: 0.571–0.812; p = 0.0118) also indicate the increase the risk of FGR development, as well as the value of the ratio of E/PLGF &gt; 138.53 (sensitivity – 100.0 %, specificity – 35.29 %) and area under the ROC-curve (AUC) 0.640 (95 % CI: 0.507– 0,759). Doppler results in pregnant women with CAH, in the presence of risk factors for FGR development, demonstrate that the CDR was not likely to differ in all groups of patients: 3.62±0.09 units in the persons with CAH and FGR (n=10) and 3.71±0.05 units in the pregnant women with CAG without FGR (n=51) in the right uterine artery; 3.63 ± 0.09 units and 3.71 ± 0.05 units in the left uterine artery, respectively (p&lt;0.05 for all values). However, if PI in the right uterine artery is more than 1.84 units and more than 1.82 units in the left uterine artery, the FGR will further develop. Conclusions. According to the results of the study, it was found that pregnant women with CAH 1-2th degrees with FGR development in the future, in the first trimester of gestation (11-12 weeks) have a decrease PlGF level, sFlt-1and the coefficient K increase. These indicators in the 11–12 weeks of pregnancy were: PlGF ≤ 8.2 pg/ml, sFlt-1 &gt; 1802.59 ng/ml, coefficient K ≥ 95.36 units, and PG/PlGF &gt; 7.21 units, E/PlGF &gt; 138.53 units and CHG/PlGF ≥ 30.14 units.In the pregnant women with CAG, in whom FGR risk will subsequently determine, the significant changes in the CDR and RI at 11-12 weeks of pregnancy are not established, but with an increase of PI more than 1.84 units in the right uterine artery and more than 1.82 units in the left uterine artery FGR will further develop.

Puerariae lobatae Radix Alleviates Pre-Eclampsia by Remodeling Gut Microbiota and Protecting the Gut and Placental Barriers

Pre-eclampsia (PE) is a serious pregnancy complication, and gut dysbiosis is an important cause of it. Puerariae lobatae Radix (PLR) is a medicine and food homologous species; however, its effect on PE is unclear. This study aimed to investigate the efficacy of PLR in alleviating PE and its mechanisms. We used an NG-nitro-L-arginine methyl ester (L-NAME)-induced PE mouse model to examine the efficacy of preventive and therapeutic PLR supplementation. The results showed that both PLR interventions alleviated hypertension and proteinuria, increased fetal and placental weights, and elevated the levels of VEGF and PlGF. Moreover, PLR protected the placenta from oxidative stress via activating the Nrf2/HO-1/NQO1 pathway and mitigated placental damage by increasing intestinal barrier markers (ZO-1, Occludin, and Claudin-1) expression and reducing lipopolysaccharide leakage. Notably, preventive PLR administration corrected gut dysbiosis in PE mice, as evidenced by the increased abundance and positive interactions of beneficial bacteria including Bifidobacterium, Blautia, and Turicibacter. Fecal microbiota transplantation confirmed that the gut microbiota partially mediated the beneficial effects of PLR on PE. Our findings revealed that modulating the gut microbiota is an effective strategy for the treatment of PE and highlighted that PLR might be used as an intestinal nutrient supplement in PE patients.

Blood biomarkers to predict the onset of pre-eclampsia: A systematic review and meta-analysis

Pre-eclampsia is one of the most common pregnancy complications, and a major cause of fetal and maternal morbidity and mortality globally. Diagnosis currently takes place in the third trimester based on clinical symptoms. This systematic review and meta-analysis sought to determine the blood biomarkers that are associated with pre-eclampsia, and in particular, the biomarkers that could predict pre-eclampsia in early pregnancy. We searched the electronic databases (Medline, Embase, Cochrane Library) from inception up to March 2022. Prospective studies with 1000 or more participants that measured blood biomarkers to predict or diagnose pre-eclampsia have been included in this systematic review. Biomarkers’ measurements were considered from the first up to the third trimester, but not during labor. Data concerning pre-eclampsia, biomarker measurements and study characteristics were extracted. Meta-analysis was performed when possible. We found a total of 43 studies (assessing 62 different biomarkers in 18,170 pregnancies, have been included in this systematic review, and a total of 6 studies (assessing 2 biomarkers have been included in the meta-analysis). Statistical analysis was performed for PlGF and sFlt-1. Mean difference in PlGF levels between pre-eclampsia and healthy pregnancies, appear to increase as the pregnancy progresses. Results of sFlt-1 meta-analysis were inconclusive. No significant publication bias was identified. This is the most comprehensive and up to date systematic review and meta-analysis on this important topic on blood biomarkers for the early prediction of pre-eclampsia. Further This research highlights the urgent needed for further discovery research to identify blood biomarkers that could predict the development of pre-eclampsia.

2, 2′, 4, 4′-tetrabromodiphenyl ether induces placental toxicity via activation of p38 MAPK signaling pathway in vivo and in vitro

2, 2', 4, 4'-tetrabromodiphenyl ether (BDE-47) is one of the most important polybrominated diphenyl ethers (PBDEs) congeners, and epidemiological studies have shown that it can cause adverse pregnancy outcomes. The aim of our study was to investigate the role of placental injury in BDE-47-induced adverse pregnancy outcomes through in vivo and in vitro models. From day 0.5 to day 16.5 of pregnancy of ICR mice, BDE-47 oral doses of 0, 25, 50 and 100mg/kg/day were administered. Immunohistochemical staining found that BDE-47 inhibited the expression of CD34 in mouse placenta, and ELISA results showed that BDE-47 reduced the levels of VEGF and PlGF in the serum of pregnant mice. Western blot assays found that the expression levels of VEGF-A and invasion-related factors were decreased in the placentas of BDE-47-treated group, which indicated that BDE-47 could impair placental angiogenesis. Furthermore, BDE-47 inhibited proliferation, increased apoptosis and autophagy, and activated p38 MAPK signaling pathway in mouse placental tissue. In vitro, HTR-8/SVneo cells were treated with 0, 5, 10, 20μM BDE-47 for 24h. Wound healing assays and Transwell assays showed that BDE-47 inhibited the migration and invasion ability of HTR-8/SVneo cells. We also found that BDE-47 inhibited the proliferation of HTR-8/SVneo cells and increased apoptosis and autophagy. BDE-47 activated p38 MAPK signaling pathway in HTR-8/SVneo cells, and inhibition of p38 MAPK signaling pathway in HTR-8/SVneo cells restored the effects caused by BDE-47. In conclusion, BDE-47 impairs placental angiogenesis by inhibiting cell migration and invasion, and induces placental toxicity by inhibiting proliferation, increasing apoptosis and autophagy. In vitro, activation of p38 MAPK signaling pathway is involved in the processes of placental injury by BDE-47.

Kisspeptin treatment improves fetal-placental development and blocks placental oxidative damage caused by maternal hypothyroidism in an experimental rat model.

Maternal hypothyroidism is associated with fetal growth restriction, placental dysfunction, and reduced kisspeptin/Kiss1R at the maternal-fetal interface. Kisspeptin affects trophoblastic migration and has antioxidant and immunomodulatory activities. This study aimed to evaluate the therapeutic potential of kisspeptin in the fetal-placental dysfunction of hypothyroid Wistar rats. Hypothyroidism was induced by daily administration of propylthiouracil. Kisspeptin-10 (Kp-10) treatment was performed every other day or daily beginning on day 8 of gestation. Feto-placental development, placental histomorphometry, and expression levels of growth factors (VEGF, PLGF, IGF1, IGF2, and GLUT1), hormonal (Dio2) and inflammatory mediators (TNFα, IL10, and IL6), markers of hypoxia (HIF1α) and oxidative damage (8-OHdG), antioxidant enzymes (SOD1, Cat, and GPx1), and endoplasmic reticulum stress mediators (ATF4, GRP78, and CHOP) were evaluated on day 18 of gestation. Daily treatment with Kp-10 increased free T3 and T4 levels and improved fetal weight. Both treatments reestablished the glycogen cell population in the junctional zone. Daily treatment with Kp-10 increased the gene expression levels of Plgf, Igf1, and Glut1 in the placenta of hypothyroid animals, in addition to blocking the increase in 8-OHdG and increasing protein and/or mRNA expression levels of SOD1, Cat, and GPx1. Daily treatment with Kp-10 did not alter the higher protein expression levels of VEGF, HIF1α, IL10, GRP78, and CHOP caused by hypothyroidism in the junctional zone compared to control, nor the lower expression of Dio2 caused by hypothyroidism. However, in the labyrinth zone, this treatment restored the expression of VEGF and IL10 and reduced the GRP78 and CHOP immunostaining. These findings demonstrate that daily treatment with Kp-10 improves fetal development and placental morphology in hypothyroid rats, blocks placental oxidative damage, and increases the expression of growth factors and antioxidant enzymes in the placenta.

An observational study of pro- and anti-angiogenic factors in hypertensive disorders of pregnancy in women of African ancestry

Hypertensive disorders in pregnancy (HDPs) are the leading cause of maternal and perinatal deaths worldwide. Despite the widely reported multisystemic pathophysiology of pre-eclampsia and other HDPs, it is unknown whether these disorders represent a continuum or separate entities making clinical diagnosis a challenge. This study aimed to investigate angiogenic, metabolic and immunoregulatory specific profiles of hypertensive and gestationally matched normotensive pregnancies. A total of 200 pregnancies from a regional hospital in South Africa, via convenience sampling, were quantitatively analysed for circulating sFlt-1; PlGF; VEGF; sENG; PAPP-A; PP13; ADAMTS 12; TGF-β1 in maternal serum samples using ELISA technique. Serum protein markers TGF-β1, sENG and PAPP-A were significantly increased (p < .05) in early-onset pre-eclampsia vs. NG1 groups. sFlt-1 was significantly higher in late-onset pre-eclampsia vs NG2 groups. The GH group showed a significant increase in TGF-β1 and PAPP-A vs. NG1 counterpart. ADAMTS12 and sENG were significantly lower in gestational hypertension vs. early-onset pre-eclampsia. No significant differences were seen in PlGF, VEGF and PP13 levels across the groups. These changes show the HDP spectrum has distinct characteristics on the angiogenic profile. Based on these results, further validation of diagnostic and prognostic biomarkers of pre-eclampsia and gestational hypertension is warranted. Impact statement What is already known on this subject? Hypertensive pregnancy disorders are a public health problem with adverse effects on both mother and neonate. The elusive pathogenesis of this syndrome combined with the late prevalence of symptoms leaves clinicians with a myriad of theories and indefinite treatments. The investigation into conventional anti-/angiogenic factors has been extensively studied in pre-eclampsia patients only. The overlapping clinical presentation of pre-eclampsia and gestational hypertension further complicates the diagnosis of disorders. What do the results of this study add? The investigation of novel angiogenic, metabolic and inflammatory markers will firstly contribute to generating a database for researchers both nationally and internationally. This combinatory triad of markers will assist in elucidating and differentiating between early- and late-onset preeclampsia versus gestational hypertension. The results of our cohort study suggest possible early diagnostic markers for pre-eclampsia and gestational hypertension. What are the implications of these findings for clinical practice and/or further research? Research in this area will contribute to an improvement in early disease management which will ultimately lead to a reduction in health care costs and mortality rate locally and globally. It will also enforce diagnostic and prognostic markers for hypertensive pregnancy diseases and warrant further investigation into the proteins primarily involved in the trophoblastic invasion. This will then clarify whether these two closely related hypertensive disorders represent a continuum or two separate entities.

Characterization of the Expression of Angiogenic Factors in Cutaneous Squamous Cell Carcinoma of Domestic Cats.

Simple SummarySquamous cell carcinoma (SCC) is a malignant skin cancer that affects domestic animal species and humans with similar characteristics. Our research seeks to understand the mechanisms by which SCC progression depends on the development of a new blood supply (angiogenesis) in the host. Here, we queried our archive of cat SCC tumor samples to measure expression of genes coding for angiogenic signaling proteins that can exist in closely related forms with distinct biological properties. We observed that, when compared to normal skin, SCC tissues contained a greater abundance of gene transcripts encoding a form of the growth factor PLGF, predicted to have an altered distribution in the body. Similarly, altered patterns of expression were observed for forms of the PLGF receptor Flt-1, which can modulate angiogenesis. Future studies will test the relationship between these gene expression changes and the severity of SCC in order to establish them as predictive biomarkers of SCC progression in individual patients.Cutaneous squamous cell carcinoma (CSCC) is a common malignant skin cancer with a significant impact on health, and it is important to determine the degree of reliance of CSCC on angiogenesis for growth and metastasis. Major regulators of angiogenesis are the vascular endothelial growth factor (VEGF) family and their associated receptors. Alternative pre-mRNA splicing produces multiple isoforms of VEGF-A and PLGF with distinct biological properties. Several studies highlight the function of VEGF-A in CSCC, but there are no studies of the different isoforms of VEGF-A and PLGF for this neoplasm. We characterized the expression of three isoforms of VEGF-A, two isoforms of PLGF, and their receptors in cat CSCC biopsies compared to normal haired skin (NHS). Although our results revealed no significant changes in transcript levels of panVEGF-A or their isoforms, the mRNA levels of PLGF I and the receptors Flt-1 and KDR were downregulated in CSCC compared to NHS. Differences were observed in ligand:receptor mRNA expression ratio, with the expression of VEGF-A relative to its receptor KDR higher in CSCC, which is consistent with our hypothesis and prior human SCC studies. Immunolocalization in tissue showed increased expression of all measured factors and receptors in tumor cells compared to NHS and surrounding vasculature. We conclude that the factors measured may play a pivotal role in CSCC growth, although further studies are needed to clarify the role of angiogenic factors in feline CSCC.

Маркери ангіоґенезу та їх роль у прогнозуванні прееклампсії при багатоплідній вагітності

Purpose - to identify changes in biochemical markers of endothelial dysfunction, preclinical manifestations of preeclampsia in pregnant women with multiple pregnancies after the use of assisted reproductive technologies. Materials and methods. A prospective study of pregnant women with dichorionic diamniotic twins after assisted reproductive technologies was conducted: 35 pregnant women with twins who were offered a developed monitoring algorithm (main (І) group) and 27 pregnant women who were monitored according to generally accepted methods of managing multiple pregnancies (control (ІІ) group). Statistical processing of the research results was carried out using standard programs “Microsoft Excel 5.0” and “Statistica 6.0”. Results. During the analysis of the main and control observation groups, an increase in the pro-angiogenic biomarker PlGF was noted up to 28 weeks (I group 604.9 (83.4-814.5) pg/ml vs. 568.6 (68.1-765.3) pg/ml in the II group). Іn the term of 26-28 weeks, in the main group, we noted a decrease in the level of PlGF below 100 pg/ml in 4 (11.4%) women, and in the II group in 6 (22.2%), however, clinical manifestations of preeclampsia, in patients, it was not noted. In the period of 32-34 weeks, the results of PlGF below the level of 100 pg/ml were obtained in 6 (17.1%) women of the I group against 7 (25.9%) of the women of the II group. In the group I we found 2 cases of moderate preeclampsia at 32 and 33 weeks of pregnancy. In the II group 4 cases of preeclampsia were identified. Two cases of moderate preeclampsia, which clinically manifested at 30 weeks. Conclusions. The anti-angiogenic factor, sFlt-1 and the pro-angiogenic factor PIGF, and their relationship are promising markers in the prediction and early diagnosis of preeclampsia in the multiple pregnancies after assisted reproductive technologies. Based on monitoring the growth dynamics of the sFlt-1/PIGF ratio, it is possible to predict the early development of preeclampsia even without clinical symptoms, and to make a decision about the possibility of extending the pregnancy or the need for urgent delivery. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

Does HAART dysregulate angiogenesis in HIV infected preeclampsia?

A dysregulation of angiogenic mediators has been implicated in HIV infection. Inconsistent data exists on highly active antiretroviral therapy (HAART) usage in pregnancy and its association with PE development. In view of the high prevalence of HIV infection and PE in SA, this study was aimed at determining PlGF and sFlt-1 levels in HIV-infected normotensive and preeclamptic pregnancies treated with HAART. Both PlGF and sFlt-1 were quantified in serum from HIV positive [normotensive (N+) and preeclamptic (P+)]; and HIV negative [normotensive (N-) and preeclamptic (P-)] pregnancies, using a Milliplex Multiplex immunoassay. sFlt-1 was significantly upregulated in P+ vs the N+ groups. PlGF was significantly downregulated in PE vs normotensive groups, regardless of HIV status. sFlt-1/PlGF ratio was significantly increased in PE- vs the N- groups. We report an amplification of sFlt-1 in lieu of PlGF down-regulation in HIV-infected pregnancies receiving HAART .

Follicular fluid PlGF and IVF/ICSI outcomes among PCOS and normo-ovulatory women using different controlled hyperstimulation protocols: A prospective case-control study

BackgroundGonadotropin-releasing hormone (GnRH) analogues are used to prevent premature luteinizing hormone (LH) surge during In-Vitro Fertilization. However, the follicular fluid levels of the Placental growth factor (FF PlGF), the novel angiogenic factor, differ significantly between GnRH-agonist and GnRH-antagonist protocols. Thus, we compared the IVF/ICSI outcomes and their correlations with FF PlGF levels in polycystic ovary syndrome (PCOS) and normo-ovulatory women during different hyperstimulation protocols. MethodsThis case-control study is a re-analysis of two prospective trials that were conducted on women who were referred to Orient Hospital, Damascus, Syria, from December 2019 to August 2021. A total of 75 PCOS-women (PCOS-Agonist, n = 53; PCOS-Antagonist, n = 22) and 83 normo-ovulatory women (Control-Agonist, n = 50; Control-Antagonist, n = 33) were included. Follicular fluid samples were collected on retrieval day. ResultsAlthough PCOS-women were stimulated using lower gonadotropin doses, the Ovarian-sensitivity-indexes were higher in PCOS-groups (PCOS-Agonist vs Control-Agonist; P-value <0.001), (PCOS-Antagonist vs Control-Antagonist; P-value = 0.042). However, FF PlGF levels, maturation rate, fertilization rate, and oocytes morphology were comparable between PCOS and controls independently of the protocol used. Interestingly, FF PlGF levels were positively correlated with Ovarian-sensitivity-indexes in the PCOS-Antagonist, Control-Agonist, and Control-Anta groups, but not in the PCOS-Agonist group. Nevertheless, FF PlGF levels were comparable between pregnant and non-pregnant women in all studied groups. ConclusionsAlthough PCOS exaggerates ovarian response to stimulation irrespective of the protocol used, it does not have a detrimental impact on oocytes morphology or competence. Moreover, FF PlGF levels could be a marker of the ovarian response other than a predictor of pregnancy achievement.

Association between placenta accreta spectrum and third-trimester serum levels of vascular endothelial growth factor, placental growth factor, and soluble Fms-like tyrosine kinase-1: A meta-analysis.

Antenatal suspicion of placenta accreta spectrum (PAS) currently relies on ultrasonographic findings, color doppler, and MRI, which have rendered it operator and expertise-dependent. No serum markers for PAS have been integrated into clinical practice yet. The aim of this meta-analysis was to identify potential serum markers for PAS by investigating third-trimester serum levels of vascular endothelial growth factor (VEFG), placental growth factor (PIGF), and soluble Fms-like tyrosine kinase-1 (sFlt-1) among PAS-cases and controls. PubMed, Scopus, EBSCO, Web of Science, and CNKI databases were systematically searched for relevant articles. Random-effects model was applied to calculate the overall standardized mean difference (SMD) for each marker. Subgroup analysis and meta-regression were performed to assess for potential covariates. Eight studies involving 366 PAS-cases and 518 controls were included. Third trimester sFlt-1 levels were significantly lower in PAS-cases when compared to controls (SMD=-7.76, 95%CI=-10.42 to -5.10). This was, to a certain extent, consistent among studies though they differed in their extent of significance. Levels of VEGF (SMD=1.59, 95%CI=-0.07 to 3.25) and PlGF (SMD=-0.49, 95%CI=-1.66 to 0.67) were not significantly different between PAS cases and controls, in which studies demonstrated conflicting results. Third trimester sFlt-1 levels may be useful to predict PAS. Nonetheless, further studies are recommended to better understand conflicting results before adopting either VEGF or PlGF.