Effects of dietary fat, nitrate, and 3-nitrooxypropanol and their combinations on methane emission, feed intake, and milk production in dairy cows.

The objective of the present study was to investigate the effect of individual and combined use of dietary fat, nitrate, and 3-nitrooxypropanol (3-NOP) on dairy cows' enteric methane (CH4) emission and production performance. Twenty-four primiparous and 24 multiparous Danish Holstein cows (111 ± 44.6 d in milk; mean ± standard deviation) were included in an incomplete 8 × 8 Latin square design with six 21-d periods. Dietary treatments were organized in a 2 × 2 × 2 factorial arrangement aiming for 2 levels of FAT (30 or 63 g of crude fat/kg of dry matter [DM]; LF or HF, respectively), 2 levels of NITRATE (0 or 10 g of nitrate/kg of DM; UREA or NIT, respectively), and 2 levels of 3-NOP (0 or 80 mg/kg DM; BLANK or NOP, respectively). Treatments were included in ad libitum-fed partial mixed rations in bins that automatically measured feed intake and eating behavior. Additional concentrate was offered as bait in GreenFeed units used for measurement of gas emission. For total DM intake (DMI), a FAT × NITRATE interaction showed that DMI, across parities and levels of 3-NOP, was unaffected by separate fat supplementation, but reduced by nitrate with 4.6% and synergistically decreased (significant 2-way interaction) with 13.0% when fat and nitrate were combined. Additionally, 3-NOP decreased DMI by 13.4% and the combination of 3-NOP with fat and nitrate decreased DMI in an additive way (no significant 3-way interaction). The decreasing effects on DMI were more pronounced in multiparous cows than in primiparous cows. For treatments with largest reductions in DMI, eating behavior was altered toward more frequent, but smaller meals, a slower eating rate and increased attempts to visit unassigned feed bins. Energy-corrected milk (ECM) yield increased by 6.3% with fat supplementation, whereas ECM yield did not differ among diets including nitrate (FAT × NITRATE interaction). Cows supplemented with 3-NOP had 9.0% lower ECM yield than cows fed no 3-NOP. Based on three 2-way interactions including FAT, NITRATE, and 3-NOP, the combined use of the additives resulted in antagonistic effects on CH4 reduction. A 6% to 7% reduction in CH4 yield (CH4/kg of DMI) could be ascribed to the effect of fat, a 12% to 13% reduction could be ascribed to the effect of nitrate and an 18% to 23% reduction could be ascribed to the effect of 3-NOP. Hence, no combinations of additives resulted in CH4 yield-reductions that were greater than what was obtained by separate supplementation of the most potent additive within the combination. The CH4 yield reduction potential of additives was similar between parities. Increased apparent total-tract digestibility of organic matter (OM) in cows fed combinations including nitrate or 3-NOP was a result of a NITRATE × 3-NOP interaction. Apparent total-tract digestibility of OM was also increased by fat supplementation. These increases reflected observed decreases in DMI. In conclusion, combined use of fat, nitrate, and 3-NOP in all combinations did not result in CH4 reductions that were greater than separate supplementation of the most potent additive within the combination (3-NOP > nitrate > fat). Additionally, separate supplementation of some additives and combined use of all additives reduced DMI.

How much can performance measures explain of the between-cow variation in enteric methane?

Enteric CH4 produced from dairy cows contributes to the emission of greenhouse gases from anthropogenic sources. Recent studies have shown that the selection of lower CH4-emitting cows is possible, but doing so would be simpler if performance measures already recorded on farm could be used, instead of measuring gas emissions from individual cows. These performance measures could be used for selection of low emitting cows. The aim of this analysis was to quantify how much of the between-cow variation in CH4 production can be explained by variation in performance measures. A dataset with 3 experiments and a total of 149 lactating dairy cows with repeated measures was used to estimate the between-cow variation (the variation between cow estimates) for performance and gas measures from GreenFeed (C-Lock, Rapid City, SD). The cow estimates were obtained with a linear mixed model with the diet within period effect as a fixed effect and the cow within experiment as a random effect. The cow estimates for CH4 production were first regressed on the performance and gas measures individually, and then performance and CO2 production measures were grouped in 3 subsets for principal component analysis and principal component regression. The variables that explained most of the between-cow variation in CH4 production were DMI (R2 = 0.44), among the performance measures, and CO2 production (R2 = 0.61), among gas measures. Grouping the measures increased the R2 to 0.53 when only performance measures were used, and to 0.66 when CO2 production was added to the significant performance measures. We found the marginal improvement to be insufficient to justify the use of grouped measures rather than an individual measure because the latter simplifies the model and avoids over-fitting. Investigation of other measures that can be explored to increase explanatory power of between-cow variation in CH4 production is briefly discussed. Finally, the use of residual CH4 as a measure for CH4 efficiency could be considered by using either DMI or CO2 production as the sole predicting variables.

Effect of carbohydrate type in silages and concentrates on feed intake, enteric methane and milk yield from dairy cows.

Dietary carbohydrate manipulation can be used to reduce enteric CH4 emission, but there is a lack of studies on the interaction of different types of carbohydrates that can affect feed intake and ruminal fermentation. Understanding this interaction is necessary to make the most out of CH4 mitigation feeding strategies using different dietary carbohydrates. The aim of this study was to test the effect on enteric CH4 emission, feed intake and milk production response when cows were fed either grass-clover (GCS) or corn silage (CS) as the sole forage source (55% of dry matter, DM), in combination with either barley (BAR) or dried beet pulp (DBP) as a concentrate (21.5% of DM). Twenty-four (half first and half second parity) cows were used in a crossover design with 2 periods of 21 d each, receiving 2 of 4 diets obtained from a 2 × 2 factorial arrangement of the experimental diet. Feed intake, CH4 emission metrics and milk production were recorded at the end of the experimental periods. The diets had NDF concentrations between 258 and 340 g/kg of DM, and starch concentrations between 340 and 7.45 g/kg of DM (CS-BAR and GCS-DBP, respectively). The effects of silage and concentrate on dry matter intake (DMI) were additive, with the highest feed intake in cows fed COR-BAR, followed by cows fed COR-DBP, GCS-BAR, and GCS-DBP (21.2, 19.9, 19.1, and 18.3 kg/d). Energy corrected milk (ECM) yield was not affected by silage source in first parity cows, but it was higher for cows fed CS than cows fed GCS in second parity. The effects of silage and concentrate on CH4 production (g/d), yield (g/kg of DMI) and intensity (g/kg of ECM) were not additive as cows fed GCS had similar responses regardless of the concentrate used, but cows fed CS had lower CH4 production, yield and intensity, when fed BAR instead of DBP. The lower CH4 production, yield and intensity in cows fed CS-BAR compared with other diets could be partially explained by the nonlinear relationship between ruminal VFA and carbohydrates (NDF and starch) concentration reported in literature, however, we observed a linear relationship between acetate:propionate ratio and CH4 yield, suggesting possible other effects. The effects of silage and concentrate on the ruminal VFA were additive in first parity cows, but not in second parity cows. The interaction between dietary CHO type and parity might indicate an effect of feed intake or the energy balance of the cow. Feeding cows silage and concentrate both rich in starch can result in the lowest enteric CH4 emission.

Women's perceptions of cardiovascular risk after preeclampsia: a qualitative study.

BACKGROUND: Preeclampsia is associated with increased risk of cardiovascular disease later in life, but studies suggest that women with previous preeclampsia are not aware of this. Little is known about how these women perceive the condition and the associated long-term risks. We examined the experiences and perceptions of preeclampsia and the increased risk of cardiovascular disease (CVD) later in life among Danish women with previous preeclampsia and their attitudes towards CVD risk screening. METHODS: Ten individual semi-structured interviews were conducted with women with previous preeclampsia. Data were analysed using thematic analysis. RESULTS: We identified six themes: 1) Experiences and perceptions of being diagnosed with preeclampsia, 2) Awareness about increased risk of CVD later in life, 3) Knowledge as a precondition for action, 4) The perception of CVD risk as being modifiable, 5) Motivators for and barriers to a healthy lifestyle, and 6) Screening for CVD. Awareness of the severity of preeclampsia was limited prior to being diagnosed. Particularly among those with few or no symptoms, preeclampsia was perceived as a non-severe condition, which was further reinforced by the experience of having received very little information. Nonetheless, some women were shocked by the diagnosis and feared for the health of the offspring. Many women also experienced physical and psychological consequences of preeclampsia. Awareness of the increased risk of later CVD was lacking; yet, when informed, the women considered this to be essential knowledge to be able to act accordingly. The risk of future CVD was perceived to be partly modifiable with a healthy lifestyle, and the women expressed a need for counselling on appropriate lifestyle changes to reduce CVD risk. Other factors were also mentioned as imperative for lifestyle changes, including social support. The women were generally positive towards potential future screening for CVD because it could provide them with information about their health condition. CONCLUSIONS: After preeclampsia, women experienced a lack of knowledge on preeclampsia and the increased risk of CVD later in life. Improved information and follow-up after preeclampsia, including guidance on CVD risk reduction and support from health professionals and family, are warranted.

Biomarkers and Their Relation to Cardiac Function Late After Peripartum Cardiomyopathy.

BACKGROUND: Angiogenic imbalance involving the placental protein soluble Fms-like tyrosine kinase-1 (sFlt-1) and cleavage of the nursing-hormone prolactin by the enzyme cathepsin D (CD) both play a role in the pathogenesis of peripartum cardiomyopathy (PPCM). We hypothesized that angiogenic imbalance and increased activity of CD have a long-lasting impact in women with PPCM. METHODS AND RESULTS: A nationwide Danish cohort of women with PPCM (PPCM group, n = 28), age matched women with previous preeclampsia (n = 28) and uncomplicated pregnancies (n = 28) participated in a follow-up study including biomarker analysis, exercise testing and cardiac magnetic resonance imaging. The median time to follow-up was 91 months (range 27-137 months) for the PPCM group. Levels of sFlt-1, placental growth factor, N-terminal pro-natriuretic brain peptide, and copeptin were all significantly higher in the PPCM group. More women in the PPCM group had detectable CD activity (68%) compared with the preeclampsia group (29%) and uncomplicated pregnancies group (36%) (P = .0002). Levels of angiogenic factors and biomarkers correlated inversely with maximal exercise capacity and cardiac functional parameters assessed with cardiac magnetic resonance imaging. CONCLUSIONS: Women with PPCM had higher biomarker levels and CD activity up to 7 years after diagnosis. Higher biomarker levels correlated inversely with maximal exercise capacity and markers of cardiac dysfunction suggesting that persistent angiogenic imbalance and increased CD activity is associated with residual cardiac dysfunction.

Degree of pregnancy planning and recommended pregnancy planning behavior among women with and without chronic medical conditions - A large hospital-based cross-sectional study.

INTRODUCTION: Pregnancy planning allows women to engage in pregnancy planning behaviors to optimize health status in the preconception period. Women with chronic medical conditions have a higher risk for adverse pregnancy outcomes and therefore preconception care is recommended. The aim was to compare pregnancy planning among women with and without chronic medical conditions, and to assess adherence to the recommended pregnancy planning behaviors on folic acid intake, physical activity and abstention from smoking and alcohol, among women with and without chronic medical conditions stratified by pregnancy planning. MATERIAL AND METHODS: A cross-sectional study with data from 28 794 pregnancies. Pregnancy planning was measured with the Swedish Pregnancy Planning Scale. Multiple Poisson regression with robust variance estimates was used to assess the associations between chronic medical condition (yes/no and main categories) and pregnancy planning, and chronic medical condition status and pregnancy planning behaviors stratified by pregnancy planning. RESULTS: In the study population, 74% reported high degree of pregnancy planning, and 22% had one or more chronic medical conditions. We found no overall association between chronic medical condition and pregnancy planning (adjusted rate ratio [RR] 1.00, 95% confidence interval [CI] 0.98-1.01). However, women with type 2 diabetes and mental illness were significantly less likely to plan their pregnancies than women without these conditions (aRR 0.73, 95% CI 0.61-0.88; aRR 0.91, 95% CI 0.87-0.96, respectively). Women with chronic medical conditions were more likely to adhere to the recommended planning behaviors; intake of folic acid, abstention from alcohol prior to pregnancy and no binge drinking in early pregnancy. CONCLUSIONS: Overall, pregnancies were highly planned. Women with chronic medical conditions did not show a higher degree of pregnancy planning than women without chronic medical conditions but were, however. more likely to adhere to the generally recommended pregnancy planning behaviors (ie intake of folic acid and abstention from alcohol intake). Only women with mental illness and type 2 diabetes reported a lower degree of pregnancy planning. It is important that we continuously address pregnancy planning and planning behaviors for both women with and women without chronic medical conditions, especially women with type 2 diabetes and mental illness.

The impact of cardiovascular diseases on maternal deaths in the Nordic countries.

INTRODUCTION: Cardiovascular diseases have become increasingly important as a cause of maternal death in the Nordic countries. This is likely to be associated with a rising incidence of pregnant women with congenital and acquired cardiac diseases. Through audits, we aim to prevent future maternal deaths by identifying causes of death and suboptimal factors in the clinical management. MATERIAL AND METHODS: Maternal deaths in the Nordic countries from 2005 to 2017 were identified through linked registers. The national audit groups performed case assessments based on hospital records, classified the cause of death, and evaluated the standards of clinical care provided. Key messages were prepared to improve treatment. RESULTS: We identified 227 maternal deaths, giving a maternal mortality rate of 5.98 deaths per 100 000 live births. The most common cause of death was cardiovascular disease (n = 36 deaths). Aortic dissection/rupture, myocardial disease, and ischemic heart disease were the most common diagnoses. In nearly 60% of the cases, the disease was not recognized before death. In more than half of the deaths, substandard care was identified (59%). In 11 deaths (31%), improvements to care that may have made a difference to the outcome were identified. CONCLUSIONS: Between 2005 and 2017, cardiovascular diseases were the most common causes of maternal deaths in the Nordic countries. There appears to be a clear potential for a further reduction in these maternal deaths. Increased awareness of cardiac symptoms in pregnant women seems warranted.

Ensiled pulp from biorefining increased milk production in dairy cows compared with grass-clover silage.

The objective of the current study was to examine the effect of fibrous pulp and partial substitution of soybean meal with green protein concentrate from biorefining of grass-clover on dry matter intake, milk production, digestibility, and eating behavior in dairy cows compared with untreated grass-clover silage and soybean meal. Biorefining of grass-clover occurred right after harvest in a production-scale twin-screw press. The twin-screw pressing separated the grass-clover into a pulp and a green juice. The green juice was fermented using lactic acid bacteria for protein precipitation and then decanted, and the precipitate was heat dried to constitute the green protein concentrate. From the same field, grass-clover was harvested 6 d later due to rainy weather and was prewilted before ensiling. The pulp and the grass-clover were ensiled in bales without additives. The production trial consisted of an incomplete 6 × 4 Latin square trial (3-wk periods; 12 wk total) including 36 lactating Holstein cows. The trial had 6 treatments in a 2 × 3 factorial design with 2 forage types (grass-clover silage and pulp silage) and 3 protein treatments (low protein, high protein with soybean meal, and high protein with a mixture of soybean meal and green protein). The trial was designed to test silage type, protein type, protein level, and the interaction between protein level and silage type. The forage:concentrate ratio was 55:45 in low protein total mixed rations (TMR) and 51:49 in high protein TMR. Low protein and high protein TMR were composed of 372 and 342 g/kg of DM of experimental silages, respectively, and green protein supplemented TMR was composed of 28.5 g/kg of DM of green protein. Silage type did not affect dry matter intake of cows. The average energy-corrected milk yield was 37.0 and 33.4 kg/d for cows fed pulp silage and grass-clover silage, respectively, resulting in an improved feed efficiency in the cows receiving pulp silage. Milk fat concentration was greater in milk from cows fed pulp silage, and milk protein concentration was lower compared with milk from cows fed grass-clover silage. The in vivo digestibility of crude protein and neutral detergent fiber was greater for pulp silage diets compared with grass-clover silage diets. Eating rate was greater, whereas daily eating duration was lower, for pulp silage diets compared with grass-clover silage diets. The partial substitution of soybean meal with green protein did not affect dry matter intake, milk yield, or eating behavior. The in vivo digestibility of crude protein in green protein supplemented diets was lower compared with soybean meal diets. The results imply that extraction of protein from grassland plants can increase the value of the fiber part of grassland plants.

Peripartum cardiomyopathy: a systematic literature review.

INTRODUCTION: Peripartum cardiomyopathy (PPCM) is a rare but potentially fatal disease defined by heart failure towards the end of pregnancy or in the months following delivery. We aim to raise awareness of the condition and give the clinician an overview of current knowledge on the mechanisms of pathophysiology, diagnostics and clinical management. MATERIAL AND METHODS: Systematic literature searches were performed in PubMed and Embase up to June 2016. Cohorts of more than 20 women with PPCM conducted after 2000 were selected to report contemporary outcomes and prognostic data. Guidelines and reviews that provided comprehensive overviews were included, too. RESULTS: New research on the pathophysiological mechanisms of PPCM points towards a two-hit multifactorial cause involving genetic factors and an antiangiogenic hormonal environment of late gestation with high levels of prolactin and sFlt-1. The prevalence of concomitant preeclampsia is high (often 30-45%) and symptoms can be similar, posing diagnostic difficulties. Most women (71-98%) present postpartum. Echocardiography is essential for diagnosis, and cardiac magnetic resonance imaging may provide new insights to pathophysiology and prognosis. Management is multidisciplinary and involves advanced heart failure therapy. Treatment, timing and mode of delivery in pregnant women depend on disease severity. The risk of relapse in subsequent pregnancies is >20%, and women are often advised against a new pregnancy. CONCLUSIONS: PPCM has a huge impact on cardiovascular health and reproductive life perspective. New insights into genetics, molecular pathophysiological mechanisms and clinical studies have resulted in potential disease-specific therapies, but many questions remain unanswered.

Changes in the pattern of paracetamol use in the periconception period in a Danish cohort.

Paracetamol is the most commonly used over-the-counter drug in pregnancy. It is generally considered to be safe, but prolonged antenatal exposure has been associated with offspring short- and long-term morbidity. Our aim was to describe the pattern of paracetamol use with a focus on frequent ingestion (more than once a week), 3 months before and in early pregnancy. In this cohort, 8650 pregnant women responded to a web-based clinical questionnaire that included questions about drug use. Paracetamol was the most used drug before and in early pregnancy (35.2% and 6.5% of respondents, respectively). The proportion of frequent users decreased from 3.9% before to 0.9% in early pregnancy. Frequent paracetamol use was associated with smoking, co-morbidities, body mass index ≥ 25 kg/m(2), unplanned pregnancy, no education and inability to understand Danish. A significant decrease in the proportion of women with any paracetamol use in early pregnancy was noted after access to large packs was restricted by legislation.

Association between Preeclampsia and Long-term Risk of Arrhythmias.

BACKGROUND: Preeclampsia (PE), a pregnancy-induced hypertensive disorder, affects 4-5% of pregnancies worldwide. It is well known that hypertension is associated with an increased risk of arrhythmias; however, data on the association between PE and arrhythmias are sparse. METHODS: In this observational cohort study, we identified all primiparous women who gave birth in Denmark (1997-2016) using Danish nationwide registries. The women were stratified on whether they developed PE during primiparous pregnancy, and followed from primiparous pregnancy to incident arrhythmia, emigration, death, or end of study (December 31, 2018). RESULTS: A total of 523,271 primiparous women with a median age of 28 years were included and 23,367 (4.5%) were diagnosed with PE. During a median follow-up of 10.1 years, women with vs without PE were associated with a higher incidence of arrhythmias (1.42% vs 1.02%): 1) Composite of cardiac arrest, ventricular tachycardia/fibrillation, or ICD implantation (adjusted HR 1.60 [95% CI 1.14-2.24]), 2) Composite of advanced 2nd degree or 3rd degree atrioventricular block, sinoatrial dysfunction, or pacemaker implantation (adjusted HR 1.48 [95% CI 0.97-2.23]), 3) Composite of supraventricular tachyarrhythmias or extra systoles (adjusted HR 1.34 [95% CI 1.19-1.51]), 4) Composite of all the above-mentioned arrhythmias (adjusted HR 1.37 [95% CI 1.23-1.54]). CONCLUSION: Preeclamptic women were associated with a significantly and at hitherto unknown long-term increased rate of arrhythmias. This finding suggests that women with PE may benefit from cardiovascular risk assessment, screening, and preventive education.

This study examined whether preeclampsia, a condition that can occur during pregnancy and cause high blood-pressure, was linked to heart rhythm problems in first-time mothers. Women who had preeclampsia in their first pregnancy were more likely to develop heart rhythm problems later in life.This suggests that women with a history of preeclampsia might need extra attention to prevent future problems.

Early-onset vs late-onset preeclampsia and risk of coronary atherosclerosis later in life: a clinical follow-up study.

BACKGROUND: Younger women with previous preeclampsia have an increased risk of coronary atherosclerosis. It is unknown if this risk is associated with the time of onset of preeclampsia. OBJECTIVE: This study aimed to investigate if women with early-onset preeclampsia have a higher risk of coronary atherosclerosis compared with women with late-onset preeclampsia, independent of other perinatal risk factors. STUDY DESIGN: A total of 911 women with previous preeclampsia aged 35 to 55 years participated in a clinical follow-up study, including clinical examination, comprehensive questionnaires, and cardiac computed tomography scan 13 years (range, 0-28) after index pregnancy. Early- and late-onset preeclampsia were defined as gestational age at delivery of <34+0 and ≥34+0 gestational weeks, respectively. The primary outcome of the study was the presence of coronary atherosclerosis on the cardiac computed tomography. A logistic regression analysis was performed to investigate the association between time of onset of preeclampsia, perinatal risk factors, and the primary outcome. RESULTS: Women with early-onset preeclampsia (N=139) were older (46.2±5.7 vs 44.4±5.5 years; P<.001), more likely to have hypertension (51.1% vs 35.1%; P≤.001), and had a higher body mass index (27.9±6.3 vs 26.9±5.5 kg/m2; P=.051) compared with women with late-onset preeclampsia (N=772) at follow-up. The prevalence of the primary outcome (coronary atherosclerosis) on the cardiac computed tomography among women with early- and late-onset preeclampsia was 28.8% vs 22.2%, respectively (P=.088; adjusted odds ratio, 1.74; 95% confidence interval, 1.01-3.01; P=.045 after adjustment for maternal age at index pregnancy, prepregnancy body mass index, parity, diabetes in pregnancy, smoking in pregnancy, offspring birthweight and sex, and follow-up length). CONCLUSION: Women with early-onset preeclampsia had a slightly higher risk of coronary atherosclerosis compared with women with late-onset preeclampsia. However, according to the current evidence, it does not seem indicated to limit screening, diagnostic, and preventive measures for cardiovascular disease only to women with early-onset preeclampsia.

Aortic dimensions in women with previous pre-eclampsia.

BACKGROUND: Pre-eclampsia is a pregnancy related disorder associated with hypertension and vascular inflammation, factors that are also involved in the pathological pathway of aortic dilatation and aneurysm development. It is, however, unknown if younger women with previous pre-eclampsia have increased aortic dimensions. We tested the hypothesis that previous pre-eclampsia is associated with increased aortic dimensions in younger women. METHODS: The study was a cross-sectional cohort study of women with previous pre-eclampsia, aged 40-55, from the PRECIOUS population matched by age and parity with women from the general population. Using contrast-enhanced CT, aortic diameters were measured in the aortic root, ascending aorta, descending aorta, at the level of the diaphragm, suprarenal aorta, and infrarenal aorta. RESULTS: 1355 women (684 with previous pre-eclampsia and 671 from the general population), with a mean (standard deviation) age of 46.9 (4.4) were included. The pre-eclampsia group had larger mean (standard deviation) aortic diameters (mm) in all measured segments from the ascending to the infrarenal aorta (ascending: 33.4 (4.0) vs. 31.4 (3.7), descending: 23.9 (2.1) vs. 23.3 (2.0), diaphragm: 20.8 (1.8) vs. 20.4 (1.8), suprarenal: 22.9 (1.9) vs. 22.0 (2.0), infrarenal: 19.3 (1.6) vs. 18.6 (1.7), p â€‹< â€‹0.001 for all, also after adjustment for age, height, parity, menopause, dyslipidemia, smoking and chronic hypertension. Guideline-defined ascending aortic aneurysms were found in 8 vs 2 women (p â€‹= â€‹0.12). CONCLUSIONS: Women with previous pre-eclampsia have larger aortic dimensions compared with women from the general population. Pre-eclampsia was found to be an independent risk factor associated with a larger aortic diameter.

Generation of minipigs with targeted transgene insertion by recombinase-mediated cassette exchange (RMCE) and somatic cell nuclear transfer (SCNT).

Targeted transgenesis using site-specific recombinases is an attractive method to create genetically modified animals as it allows for integration of the transgene in a pre-selected transcriptionally active genomic site. Here we describe the application of recombinase-mediated cassette exchange (RMCE) in cells from a Göttingen minipig with four RMCE acceptor loci, each containing a green fluorescence protein (GFP) marker gene driven by a human UbiC promoter. The four RMCE acceptor loci segregated independent of each other, and expression profiles could be determined in various tissues. Using minicircles in RMCE in fibroblasts with all four acceptor loci and followed by SCNT, we produced piglets with a single copy of a transgene incorporated into one of the transcriptionally active acceptor loci. The transgene, consisting of a cDNA of the Alzheimer's disease-causing gene PSEN1M146I driven by an enhanced human UbiC promoter, had an expression profile in various tissues similar to that of the GFP marker gene. The results show that RMCE can be done in a pre-selected transcriptionally active acceptor locus for targeted transgenesis in pigs.

Acupuncture and/or sweeping of the fetal membranes before induction of labor: a prospective, randomized, controlled trial.

AIMS: To evaluate the efficacy of acupuncture, and sweeping of the fetal membranes, as methods for induction of labor. METHODS: Four hundred and seven pregnant women with normal singleton pregnancies and cephalic presentations were randomized at three delivery wards in Denmark at day 290 of gestation into groups of acupuncture, sweeping, acupuncture and sweeping and controls. The primary objective was to compare the proportion of women going into labor before induction of labor at 294 days in the four groups. The secondary objective was to compare the combined groups: with and without acupuncture, and with and without sweeping of the fetal membranes. The midwives, completing the forms for the trial at labor or induction, were blinded to group assessments. RESULTS: Four hundred and seventeen women were randomized. Ten were excluded after randomization. One hundred and four women were randomized to acupuncture, 103 to sweeping of the membranes, 100 to both acupuncture and sweeping, and 100 were randomized to the control group. Comparison of the four groups demonstrated no significant difference in the number of women achieving spontaneous labor before planned induction. No difference was demonstrated by comparing the combined groups treated with acupuncture with the groups not treated with acupuncture (P=0.76). However, significantly more women went into labor before planned induction (P=0.02) in the combined groups receiving sweeping, compared with the groups not treated with sweeping. CONCLUSIONS: Acupuncture at 41+ weeks of gestation did not reduce the need for induction. The study was of a sufficient size to demonstrate, in parallel, that sweeping of the fetal membranes significantly reduced the need of induction, sparing about 15% for formal induction of labor.

Pre-eclampsia in a first pregnancy and subsequent pregnancy outcomes: a nationwide cohort study.

BACKGROUND: For women whose first pregnancy was complicated by pre-eclampsia (PE), particularly if severe and requiring early birth, the risk of recurrence and maternal and neonatal outcomes at subsequent birth are important considerations. METHODS: In this observational cohort study, all primiparous women who gave birth in Denmark between 1997 and 2016 were identified using nationwide registries. Women were stratified by whether they developed PE and followed from date of birth until subsequent birth, emigration, death or end of study (December 2016). The cumulative incidences of subsequent birth among women with versus without PE were assessed using the Aalen-Johansen estimator. Subsequent outcomes including PE recurrence and maternal and neonatal morbidity and mortality were also examined. Factors associated with subsequent birth and recurrent PE were examined using multivariable Cox regression models. RESULTS: Among 510 615 primiparous women with singleton pregnancies, 21 683 (4.2%) developed PE, with 1819 (0.4%) being early-onset PE (birth <34 weeks). Women with PE had a lower subsequent birth rate (57.4%) compared with women without PE (61.2%), and it was considerably lower among women with early-onset PE (49.4%). Among women with PE who had a subsequent birth, the overall recurrence rate of PE was 15.8% and higher among those with early-onset PE (31.5%). The gestational age increased with a median of 3 days (IQR -5 to 14) overall and 50 days (IQR 35-67) among those with early-onset PE. Moreover, neonatal and maternal morbidity and mortality were substantially improved in a subsequent pregnancy. CONCLUSIONS: Primiparous women with PE have a significantly lower rate of a subsequent birth than women without PE, yet the absolute difference was modest. Although the overall risk of recurrent PE is 1 in 6, maternal and neonatal morbidity and mortality at subsequent birth are substantially improved.

Preeclampsia and Long-Term Risk of Venous Thromboembolism.

Importance: As venous thromboembolism (VTE) remains one of the leading causes of maternal mortality, identifying women at increased risk of VTE is of great importance. Preeclampsia is a pregnancy-induced hypertensive disorder with generalized endothelial dysfunction. Some studies suggest that preeclampsia is associated with an increased risk of VTE, but much controversy exists. Objective: To examine the association between preeclampsia and the risk of VTE during pregnancy, during the puerperium, and after the puerperium. Design, Setting, and Participants: This observational cohort study used Danish nationwide registries to identify all eligible primiparous women who gave birth in Denmark from January 1, 1997, to December 31, 2016. The women were followed up from primiparous pregnancy to incident VTE, emigration, death, or the end of the study (December 31, 2016). Statistical analyses were carried out from January to May 2023. Exposure: Preeclampsia during primiparous pregnancy. Main Outcomes and Measure: The main outcome was incident VTE, and the secondary outcome was all-cause mortality. Results: A total of 522 545 primiparous women (median age, 28 years [IQR, 25-31 years]) were included, and 23 330 (4.5%) received a diagnosis of preeclampsia. Women with preeclampsia were of similar age to women without preeclampsia but had a higher burden of comorbidities. During a median follow-up of 10.2 years (IQR, 5.2-15.4 years), preeclampsia was associated with a higher incidence of VTE compared with no preeclampsia (incidence rate, 448.8 [95% CI, 399.9-503.5] vs 309.6 [95% CI, 300.6-319.9] per 1000 patient-years, corresponding to an unadjusted hazard ratio [HR] of 1.45 [95% CI, 1.29-1.63] and an adjusted HR of 1.43 [95% CI, 1.27-1.61]). When stratified according to the subcategories of VTE, preeclampsia was associated with an increased rate of deep vein thrombosis (unadjusted HR, 1.51 [95% CI, 1.32-1.72] and adjusted HR, 1.49 [95% CI, 1.31-1.70]) as well as pulmonary embolism (unadjusted HR, 1.39 [95% CI, 1.09-1.76]; adjusted HR, 1.36 [95% CI, 1.08-1.73]). These findings held true in landmark analyses during pregnancy, during the puerperium, and after the puerperium. Conclusions and Relevance: This cohort study suggests that preeclampsia was associated with a significantly increased risk of VTE during pregnancy, during the puerperium, and after the puerperium, even after thorough adjustment. Future studies should address how to improve the clinical management of women with a history of preeclampsia to prevent VTE.

Clinical skills training in obstetrics - a descriptive survey of current practice in Denmark.

The Danish National Board of Health has recommended that labor wards establish regular obstetric emergency skills training programs. The aim of this study was to describe current practice in Denmark. A questionnaire was sent to all obstetric departments in Denmark in 2008. All responded. Simulation-based training was conducted in 26/28 obstetrical departments. Settings for the training programs were mainly local. Training was provided for shoulder dystocia, postpartum bleeding and basic neonatal resuscitation in almost all the departments, but was not organized in a uniform way. Neither the program itself nor the participants' performance was evaluated in a structured or validated way. Obstetric emergency skills training is being actively conducted in the majority of the Danish labor wards. However, it still remains a challenge to ensure the uniform organization and evaluation of the current training programs. Development of validated national or international standards for an obstetric training program should be a future aim.

Vacuum extraction: development and test of a procedure-specific rating scale.

OBJECTIVES: To develop and validate an Objective Structured Assessment of Technical Skills (OSATS) scale for vacuum extraction. DESIGN: Two-part study design: Primarily, development of a procedure-specific checklist for vacuum extraction. Hereafter, validation of the developed OSATS scale for vacuum extraction in a prospective observational study. SETTING: Rigshospitalet, University Hospital of Copenhagen. POPULATION: For development, an obstetric expert from each labor ward in Denmark (28 departments) was invited to participate. For validation, nine first-year residents and 10 chief physicians with daily work in the obstetric field were tested. METHODS: The Delphi method was used for development of the scale. In a simulated vacuum extraction scenario, first-year residents and obstetric chief physicians were rated using the developed OSATS scale for vacuum extraction to test construct validity of the scale. MAIN OUTCOME MEASURES: Consensus for the content of the scale. To test the scale of Cronbach's alpha, interclass correlation and differential item function was calculated in the prospective study. RESULTS: 89% completed the first and 61% completed the second Delphi round. Hereafter, consensus was obtained. There was a significant difference between residents' and experts' performance for total score and for the score of the separate parts of the scale. Cronbach's alpha for total score and for the separate parts of the scale was 0.91-0.95 and interclass correlation 0.84-0.9. CONCLUSIONS: The OSATS scale for vacuum extraction is a reliable test for differentiating between competence levels in a simulated setting.