Placental size at gestational week 36: Comparisons between ongoing pregnancies and deliveries.

INTRODUCTION: We aimed to compare placental size and placental size relative to fetal size (ratio) in ongoing pregnancies examined by magnetic resonance imaging (MRI) at gestational week 36 with placental size among all deliveries at gestational week 36 during the same time period. MATERIAL AND METHODS: Ongoing unselected singleton pregnancies (n = 89) were examined by MRI at median gestational week 36+5 days during 2017-2018, and placental and fetal volumes (cm3 ) were calculated. The placental size and ratio in ongoing pregnancies were compared with placental size and ratio among all deliveries in Norway at gestational week 36 (median gestational week 36+4 days) during 2016-2019 (n = 5582). For comparison of size, we converted volume (cm3 ) in ongoing pregnancies into grams as: cm3 × 1.05 (density of placental and fetal tissue). RESULTS: In ongoing pregnancies, median placental size was 873 (interquartile range [IQR] 265) grams and median size of all delivered placentas was 613 (IQR 290) grams. Placental size was smaller among the delivered placentas independent of delivery mode: 760 (IQR 387) grams among elective cesarean deliveries (n = 465) and 590 (IQR 189) grams among vaginal deliveries after spontaneous onset of labor (n = 2478). Median ratio in ongoing pregnancies was higher than among deliveries: 0.31 (IQR 0.08) vs 0.21 (IQR 0.08). The ratio was higher in ongoing pregnancies independent of delivery mode: 0.24 (IQR 0.17) among elective cesarean deliveries vs 0.21 (IQR 0.05) among vaginal deliveries after spontaneous onset of labor. CONCLUSIONS: The placenta is larger in ongoing pregnancies than among deliveries. This finding suggests that placental size decreases during labor and delivery, possibly by transfer of blood to the fetus. Our finding also suggests that reference values of placental size based on delivered placentas are not valid for ongoing pregnancies.

Prehospital Blood Pressure and Clinical and Radiological Outcomes in Acute Spontaneous Intracerebral Hemorrhage.

BACKGROUND: High blood pressure (BP) is associated with poor outcome in acute spontaneous intracerebral hemorrhage. Little is known about the predictive value of prehospital BP in intracerebral hemorrhage. We aimed to investigate the relationship between prehospital BP and clinical and radiological outcomes. METHODS: This is a retrospective, hospital-based study of all adult intracerebral hemorrhage patients admitted within 24 hours of symptom onset to a large primary stroke centre during 2012 to 2019. The first prehospital and on-admission BP were recorded as systolic BP, diastolic BP, mean arterial pressure, and pulse pressure. The absolute differences between prehospital and on-admission BP were calculated (BPchange). Primary outcomes were in-hospital death, early neurological deterioration, and hematoma expansion. Associations between prehospital BP, BPchange, and outcomes were explored by regression with adjustment for relevant confounders. RESULTS: We included 426 patients aged median 76 (interquartile range 67-85) years and 203 (48%) were female. Median prehospital systolic BP was 179 (interquartile range 158-197) and diastolic BP was 100 (interquartile range 86-112) mm Hg. In-hospital death occurred in 121/426 (28%), early neurological deterioration in 107/295 (36%), and hematoma expansion in 50/185 (27%) patients. There were linear associations between 5 mm Hg increment of prehospital systolic BP (odds ratio 1.06, [95% CI, 1.01-1.12]) and mean arterial pressure (odds ratio 1.08, [95% CI, 1.01-1.15]) and in-hospital death, and between 5 mm Hg increment of prehospital diastolic BP (odds ratio 1.10, [95% CI, 1.00-1.21]) and mean arterial pressure (odds ratio 1.09, [95% CI, 1.00-1.18]) and hematoma expansion. There was a nonlinear association between prehospital systolic BP and in-hospital death. No consistent associations between prehospital BPchange and outcomes were found. CONCLUSIONS: In patients with acute intracerebral hemorrhage, elevated prehospital BP parameters were associated with in-hospital death and hematoma expansion. Changes in prehospital BP were not consistently associated with outcome. A possible U-shaped association between prehospital BP and in-hospital death needs further investigation.

Placental volume in gestational week 27 measured by three-dimensional ultrasound and magnetic resonance imaging.

INTRODUCTION: Ultrasound is the diagnostic tool of choice in pregnancy. We lack valid ultrasound methods for placental size measurements. Our aim was therefore to compare three-dimensional (3D) ultrasound with magnetic resonance imaging (MRI) for measurements of placental volume. MATERIAL AND METHODS: We measured placental volume by 3D ultrasound and MRI in 100 unselected pregnancies at 27 weeks of gestation (25+4 -28+4  weeks). The 3D ultrasound acquisitions were analyzed offline, and the placental outline was manually traced using the virtual organ computer-aided analysis (VOCAL) 30° rotational technique. The MRI examinations included a T2-weighted gradient echo sequence in the sagittal plane, with 5-mm slices through the entire uterus. The placental outline was manually traced in each slice. The correlation between 3D ultrasound and MRI placental volumes was estimated by intraclass correlation coefficients. Bland-Altman analysis was applied to visualize systematic bias and limits of agreement, in which the ratio MRI placental volume/3D ultrasound placental volume was plotted against the average of the two methods. RESULTS: The intraclass correlation coefficient between 3D ultrasound and MRI measurements was 0.49 (95% confidence interval 0.33-0.63). In general, 3D ultrasound measured smaller placental volumes (median 373 cm3 , interquartile range 309-434 cm3 ) than MRI (median 507 cm3 , interquartile range 429-595 cm3 ) and the systematic bias was 1.44. The 95% limits of agreement between the two methods were wide (0.68-2.21). CONCLUSIONS: We found poor to moderate correlation between 3D ultrasound and MRI placental volume measurements. Generally, 3D ultrasound measured smaller placental volumes than MRI, suggesting that 3D ultrasound failed to visualize the entire placenta. Our findings may hopefully contribute to the improvement of ultrasound methods for placental measurements.

Connective tissue growth factor and bone morphogenetic protein 2 are induced following myocardial ischemia in mice and humans.

We aimed to study the cardiac expression of bone morphogenetic protein 2, its receptor 1 b, and connective tissue growth factor, factors implicated in cardiac embryogenesis, following ischemia/hypoxia, heart failure, and in remodeling hearts from humans and mice. Biopsies from the left ventricle of patients with end-stage heart failure due to dilated cardiomyopathy or coronary artery disease were compared with donor hearts and biopsies from patients with normal heart function undergoing coronary artery bypass grafting. Mouse model of post-infarction remodeling was made by permanent ligation of the left coronary artery. Hearts were analyzed by real-time polymerase chain reaction and Western blotting after 24 hours and after 2 and 4 weeks. Patients with dilated cardiomyopathy and mice post-infarction had increased cardiac expression of connective tissue growth factor. Bone morphogenetic protein 2 was increased in human hearts failing due to coronary artery disease and in mice post-infarction. Gene expression of bone morphogenetic protein receptor 1 beta was reduced in hearts of patients with failure, but increased two weeks following permanent ligation of the left coronary artery in mice. In conclusion, connective tissue growth factor is upregulated in hearts of humans with dilated cardiomyopathy, bone morphogenetic protein 2 is upregulated in remodeling due to myocardial infarction while its receptor 1 b in human failing hearts is downregulated. A potential explanation might be an attempt to engage regenerative processes, which should be addressed by further, mechanistic studies.

Absence of the inflammasome adaptor ASC reduces hypoxia-induced pulmonary hypertension in mice.

Pulmonary hypertension is a serious condition that can lead to premature death. The mechanisms involved are incompletely understood although a role for the immune system has been suggested. Inflammasomes are part of the innate immune system and consist of the effector caspase-1 and a receptor, where nucleotide-binding oligomerization domain-like receptor pyrin domain-containing 3 (NLRP3) is the best characterized and interacts with the adaptor protein apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC). To investigate whether ASC and NLRP3 inflammasome components are involved in hypoxia-induced pulmonary hypertension, we utilized mice deficient in ASC and NLRP3. Active caspase-1, IL-18, and IL-1ß, which are regulated by inflammasomes, were measured in lung homogenates in wild-type (WT), ASC(-/-), and NLRP3(-/-) mice, and phenotypical changes related to pulmonary hypertension and right ventricular remodeling were characterized after hypoxic exposure. Right ventricular systolic pressure (RVSP) of ASC(-/-) mice was significantly lower than in WT exposed to hypoxia (40.8 ± 1.5 mmHg vs. 55.8 ± 2.4 mmHg, P < 0.001), indicating a substantially reduced pulmonary hypertension in mice lacking ASC. Magnetic resonance imaging further supported these findings by demonstrating reduced right ventricular remodeling. RVSP of NLRP3(-/-) mice exposed to hypoxia was not significantly altered compared with WT hypoxia. Whereas hypoxia increased protein levels of caspase-1, IL-18, and IL-1ß in WT and NLRP3(-/-) mice, this response was absent in ASC(-/-) mice. Moreover, ASC(-/-) mice displayed reduced muscularization and collagen deposition around arteries. In conclusion, hypoxia-induced elevated right ventricular pressure and remodeling were attenuated in mice lacking the inflammasome adaptor protein ASC, suggesting that inflammasomes play an important role in the pathogenesis of pulmonary hypertension.

Two-dimensional (2D) placental ultrasound measurements - The correlation with placental volume measured by magnetic resonance imaging (MRI).

INTRODUCTION: Information about placental size in ongoing pregnancies may aid the identification of pregnancies with increased risk of adverse outcome. Placental volume can be measured using magnetic resonance imaging (MRI). However, this method is not universally available in antenatal care. Ultrasound is the diagnostic tool of choice in pregnancy. Therefore, we studied whether simple two-dimensional (2D) ultrasound placental measurements were correlated with placental volume measured by MRI. METHODS: We examined a convenience sample of 104 ongoing pregnancies at gestational week 27, using both ultrasound and MRI. The ultrasound measurements included placental length, width and thickness. Placental volume was measured using MRI. The correlation between each 2D placental ultrasound measurement and placental volume was estimated by applying Pearson's correlation coefficient (r). RESULTS: Mean placental length was 17.2 cm (SD 2.1 cm), mean width was 14.7 cm (SD 2.1 cm), and mean thickness was 3.2 cm (SD 0.6 cm). Mean placental volume was 536 cm3 (SD 137 cm3). The 2D ultrasound measurements showed poor correlation with placental volume (placental length; r = 0.27, width; r = 0.37, and thickness r = 0.13). DISCUSSION: Simple 2D ultrasound measurements of the placenta were poorly correlated with placental volume and cannot be used as proximate measures of placental volume. Our finding may be explained by the large variation between pregnancies in intrauterine placental shape.

Placental size at gestational week 27 and 37: The associations with pulsatility index in the uterine and the fetal-placental arteries.

INTRODUCTION: Fetal growth restriction is known to be related to decreased fetal and placental blood flow. It is not known, however, whether placental size is related to fetal and placental blood flow. We studied the correlations of intrauterine placental volume and placental-fetal-ratio with pulsatility index (PI) in the uterine arteries, fetal middle cerebral artery, and umbilical artery. METHODS: We followed a convenience sample of 104 singleton pregnancies, and we measured placental and fetal volumes using magnetic resonance imaging (MRI) at gestational week 27 and 37 (n = 89). Pulsatility index (PI) was measured using Doppler ultrasound. We calculated cerebroplacental ratio as fetal middle cerebral artery PI/umbilical artery PI and placental-fetal-ratio as placental volume (cm3)/fetal volume (cm3). RESULTS: At gestational week 27, placental volume was negatively correlated with uterine artery PI (r = -0.237, p = 0.015, Pearson's correlation coefficient), and positively correlated with fetal middle cerebral artery PI (r = 0.247, p = 0.012) and cerebroplacental ratio (r = 0.208, p = 0.035). Corresponding correlations for placental-fetal-ratio were -0.273 (p = 0.005), 0.233 (p = 0.018) and 0.183 (p = 0.064). Umbilical artery PI was not correlated with placental volume. At gestational week 37, we found weaker and no significant correlations between placental volume and the pulsatility indices. CONCLUSIONS: Our results suggest that placental size is correlated with placental and fetal blood flow at gestational week 27.

Aquaporin-1 in cardiac endothelial cells is downregulated in ischemia, hypoxia and cardioplegia.

Aquaporin-1 (AQP1) is expressed in human and mouse hearts, but little is known about its cellular and subcellular localization and regulation. The aim of this study was to investigate the localization of AQP1 in the mouse heart and to determine the effects of ischemia and hypoxia on its expression. Mouse myocardial cells were freshly isolated and split into cardiomyocyte and non-cardiomyocyte fractions. Isolated, Langendorff-perfused C57Bl6 mouse hearts (n=46) were harvested with no intervention, subjected to 35min of ischemia or ischemia followed by 60min of reperfusion. Eleven mouse hearts were perfusion-fixed for electron microscopy. Forty C57Bl6 mice were exposed to normobaric hypoxia for one or two weeks (n=12). Needle biopsies of human left ventricular myocardium were sampled (n=30) during coronary artery bypass surgery before cardioplegia and after 30min of reperfusion. Human umbilical vein endothelial cells (HUVECs) were subjected to 4h of hypoxia with reoxygenation for either 4 or 24h. AQP1 expression was studied by electron microscopy with immunogold labeling, Western blot, and qPCR. Expression of miR-214 and miR-320 in HUVECs with hypoxia was studied with qPCR. HUVECs were then transfected with precursors and inhibitors of miR-214. AQP1 expression was confined to cardiac endothelial cells, with no signal in cardiomyocytes or cardiac fibroblasts. Immunogold electron microscopy showed AQP1 expression in endothelial caveolae with equal distribution along the basal and apical membranes. Ischemia and reperfusion tended to decrease AQP1 mRNA expression in mouse hearts by 37±9% (p=0.06), while glycosylated AQP1 protein was reduced by 16±9% (p=0.03). No difference in expression was found between ischemia alone and ischemia-reperfusion. In human left ventricles AQP1 mRNA expression was reduced following cardioplegia and reperfusion (p=0.008). Hypoxia in mice reduced AQP1 mRNA expression by 20±7% (p<0.0001), as well as both glycosylated (-47±10%, p=0.03) and glycan-free protein (-34±16%, p=0.05). Hypoxia and reoxygenation in HUVECs downregulated glycan-free AQP1 protein (-34±24%, p=0.04) and upregulated miR-214 (+287±52%, p<0.05). HUVECs transfected with anti-miR-214 had increased glycosylated (1.5 fold) and glycan-free (2 fold) AQP1. AQP1 in mouse hearts is localized to endothelial cell membranes and caveolae. Cardioplegia, ischemia and hypoxia decrease AQP1 mRNA as well as total protein expression and glycosylation, possibly regulated by miR-214.

Automatic placental and fetal volume estimation by a convolutional neural network.

INTRODUCTION: We aimed to develop an artificial intelligence (AI) deep learning algorithm to efficiently estimate placental and fetal volumes from magnetic resonance (MR) scans. METHODS: Manually annotated images from an MRI sequence was used as input to the neural network DenseVNet. We included data from 193 normal pregnancies at gestational week 27 and 37. The data were split into 163 scans for training, 10 scans for validation and 20 scans for testing. The neural network segmentations were compared to the manual annotation (ground truth) using the Dice Score Coefficient (DSC). RESULTS: The mean ground truth placental volume at gestational week 27 and 37 was 571 cm3 (Standard Deviation (SD) 293 cm3) and 853 cm3 (SD 186 cm3), respectively. Mean fetal volume was 979 cm3 (SD 117 cm3) and 2715 cm3 (SD 360 cm3). The best fitting neural network model was attained at 22,000 training iterations with mean DSC 0.925 (SD 0.041). The neural network estimated mean placental volumes at gestational week 27-870 cm3 (SD 202 cm3) (DSC 0.887 (SD 0.034), and to 950 cm3 (SD 316 cm3) at gestational week 37 (DSC 0.896 (SD 0.030)). Mean fetal volumes were 1292 cm3 (SD 191 cm3) and 2712 cm3 (SD 540 cm3), with mean DSC of 0.952 (SD 0.008) and 0.970 (SD 0.040). The time spent for volume estimation was reduced from 60 to 90 min by manual annotation, to less than 10 s by the neural network. CONCLUSION: The correctness of neural network volume estimation is comparable to human performance; the efficiency is substantially improved.

Antithrombotic Treatment, Prehospital Blood Pressure, and Outcomes in Spontaneous Intracerebral Hemorrhage.

Background In acute intracerebral hemorrhage, both elevated blood pressure (BP) and antithrombotic treatment are associated with poor outcome. Our aim was to explore interactions between antithrombotic treatment and prehospital BP. Methods and Results This observational, retrospective study included adult patients with spontaneous intracerebral hemorrhage diagnosed by computed tomography within 24 hours, admitted to a primary stroke center during 2012 to 2019. The first recorded prehospital/ambulance systolic and diastolic BP were analyzed per 5 mm Hg increment. Clinical outcomes were in-hospital mortality, shift on the modified Rankin Scale at discharge, and mortality at 90 days. Radiological outcomes were initial hematoma volume and hematoma expansion. Antithrombotic (antiplatelet and/or anticoagulant) treatment was analyzed both together and separately. Modification of associations between prehospital BP and outcomes by antithrombotic treatment was explored by multivariable regression with interaction terms. The study included 200 women and 220 men, median age 76 (interquartile range, 68-85) years. Antithrombotic drugs were used by 252 of 420 (60%) patients. Compared with patients without, patients with antithrombotic treatment had significantly stronger associations between high prehospital systolic BP and in-hospital mortality (odds ratio [OR], 1.14 versus 0.99, P for interaction 0.021), shift on the modified Rankin Scale (common OR, 1.08 versus 0.96, P for interaction 0.001), and hematoma volume (coef. 0.03 versus -0.03, P for interaction 0.011). Conclusions In patients with acute, spontaneous intracerebral hemorrhage, antithrombotic treatment modifies effects of prehospital BP. Compared with patients without, patients with antithrombotic treatment have poorer outcomes with higher prehospital BP. These findings may have implications for future studies on early BP lowering in intracerebral hemorrhage.

Aquaporin-4 in the heart: expression, regulation and functional role in ischemia.

Aquaporins (AQPs) are channel-forming membrane proteins highly permeable to water. AQP4 is found in mammalian hearts; however, its expression sites, regulation and function are largely unknown. The aim was to investigate cardiac AQP4 expression in humans and mice, its regulation by ischemia and hypoxia, and in particular its role in cardiac ischemic injury using AQP4 knockout (KO) mice. Comparable levels of AQP4 were detected by Western blot and qPCR in biopsies from human donor hearts and wild type C57Bl6 mouse hearts. In mice, AQP4 was expressed on cardiomyocyte plasmalemma (qPCR, Western blot, immunogold), and its mRNA decreased following ischemia/reperfusion (isolated hearts, p = 0.02) and after normobaric hypoxia in vivo (oxygen fraction 10 % for 1 week, p < 0.001). Isolated hearts from AQP4 KO mice undergoing global ischemia and reperfusion had reduced infarct size (p = 0.05) and attenuated left ventricular end-diastolic pressure during reperfusion (p = 0.04). Infarct size was also reduced in AQP4 KO mice 24 h after left coronary artery ligation in vivo (p = 0.036). AQP4 KO hearts had no compensatory change in AQP1 protein expression. AQP4 KO cardiomyocytes were partially resisted to hypoosmotic stress in the presence of hypercontracture. AQP4 is expressed in human and mouse hearts, in the latter confined to the cardiomyocyte plasmalemma. AQP4 mRNA expression is downregulated by hypoxia and ischemia. Deletion of AQP4 is protective in acute myocardial ischemia-reperfusion, and this molecule might be a future target in the treatment of acute myocardial infarction.

IL-18 and IL-12 synergy induces matrix degrading enzymes in the lung.

Interleukin (IL)-18 is a pro-inflammatory cytokine suggested to be involved in the development of pulmonary emphysema and inflammation. Studies involving immunology and cancer have revealed that IL-18 can have synergistic effects with IL-12. We have studied the presence of IL-18 and IL-12 receptors (IL-18R/IL-12R) in the lungs and whether IL-18 and IL-12, alone or in combination, have the ability to initiate the induction of mediators related to the development of emphysema and inflammation. The expression of the IL-18R was abundant in lungs compared to other organs (heart, liver, and spleen), and the IL-12R was also expressed in lung tissue. Mice treated with i.p. injection of recombinant murine IL-18 or IL-12 expressed significantly higher pulmonary mRNA levels of the matrix degrading enzymes metalloproteinase (MMP) 12 and cathepsin S, in addition to interferon-γ, tumor necrosis factor-α, and CXC chemokine ligand 9 (CXCL9) (all P < .05) than controls (received PBS). Treatment with IL-18 and IL-12 in combination showed an even more pronounced induction of these mediators, as well as a significant increase in MMP-9, IL-6, IL-1ß, and transforming growth factor-ß (P < .05). Furthermore, cellular apoptosis in lung tissue was induced. Immunohistochemical analysis revealed T-cell infiltration in pulmonary vessels following co-stimulation. In summary, IL-18 and IL-12 exert a synergistic effect on the lungs by inducing MMPs, cathepsins S, and pro-inflammatory cytokines, which may promote pulmonary emphysema and inflammation. The synergy between IL-18 and IL-12 involves infiltration of T-cells in the lungs, possibly induced by the T-cell chemoattractant CXCL9.

Long-term levosimendan treatment improves systolic function and myocardial relaxation in mice with cardiomyocyte-specific disruption of the Serca2 gene.

In human heart failure (HF), reduced cardiac function has, at least partly, been ascribed to altered calcium homeostasis in cardiomyocytes. The effects of the calcium sensitizer levosimendan on diastolic dysfunction caused by reduced removal of calcium from cytosol in early diastole are not well known. In this study, we investigated the effect of long-term levosimendan treatment in a murine model of HF where the sarco(endo)plasmatic reticulum ATPase (Serca) gene is specifically disrupted in the cardiomyocytes, leading to reduced removal of cytosolic calcium. After induction of Serca2 gene disruption, these mice develop marked diastolic dysfunction as well as impaired contractility. SERCA2 knockout (SERCA2KO) mice were treated with levosimendan or vehicle from the time of KO induction. At the 7-wk end point, cardiac function was assessed by echocardiography and pressure measurements. Vehicle-treated SERCA2KO mice showed significantly diminished left-ventricular (LV) contractility, as shown by decreased ejection fraction, stroke volume, and cardiac output. LV pressure measurements revealed a marked increase in the time constant (τ) of isovolumetric pressure decay, showing impaired relaxation. Levosimendan treatment significantly improved all three systolic parameters. Moreover, a significant reduction in τ toward normalization indicated improved relaxation. Gene-expression analysis, however, revealed an increase in genes related to production of the ECM in animals treated with levosimendan. In conclusion, long-term levosimendan treatment improves both contractility and relaxation in a heart-failure model with marked diastolic dysfunction due to reduced calcium transients. However, altered gene expression related to fibrosis was observed.