Pre-eclampsia screening in Denmark (PRESIDE): national validation study.

OBJECTIVES: To investigate the predictive performance of the Fetal Medicine Foundation (FMF) first-trimester screening algorithm for pre-eclampsia in a Danish population and compare screening performance with that of the current Danish strategy, which is based on maternal risk factors. METHODS: This was a prospective study of women with a singleton pregnancy attending for their first-trimester ultrasound scan and screening for aneuploidies at six Danish university hospitals between May 2019 and December 2020. Prenatal data on maternal characteristics and medical history were recorded, and measurements of mean arterial pressure (MAP), uterine artery pulsatility index (UtA-PI), serum pregnancy-associated plasma protein-A (PAPP-A) and serum placental growth factor (PlGF) were collected without performing a risk assessment for pre-eclampsia. Information on acetylsalicylic acid use was recorded. After delivery, pregnancy outcome, including gestational age at delivery and pre-eclampsia diagnosis, was recorded. Pre-eclampsia risk assessment for each woman was calculated blinded to outcome using the FMF screening algorithm following adjustment to the Danish population. Detection rates (DRs) of the FMF algorithm were calculated for a fixed screen-positive rate (SPR) of 10% and for the SPR achieved in the current Danish screening. RESULTS: A total of 8783 pregnant women were included, with a median age of 30.8 (interquartile range (IQR), 28.1-33.9) years. The majority were white (95%), naturally conceiving (90%), non-smokers (97%) and had no family history of pre-eclampsia (96%). The median body mass index was 23.4 (IQR, 21.2-26.6) kg/m2 . A complete risk assessment including maternal characteristics, MAP, UtA-PI, PlGF and PAPP-A was available for 8156 women (92.9%). In these women, UtA-PI was measured bilaterally with a median value of 1.58 (IQR, 1.27-1.94) and the median resting MAP of 80.5 (IQR, 76.1-85.4) mmHg in two consecutive measurements. Among these, 303 (3.7%) developed pre-eclampsia, including 55 (0.7%) cases of pre-eclampsia with delivery < 37 weeks of gestation and 16 (0.2%) cases of pre-eclampsia with delivery < 34 weeks. At a SPR of 10%, combined screening using the FMF algorithm based on maternal characteristics, MAP, UtA-PI, PlGF and PAPP-A had a DR of 77.4% (95% CI, 57.6-97.2%) for pre-eclampsia with delivery < 34 weeks, 66.8% (95% CI, 54.4-79.1%) for pre-eclampsia with delivery < 37 weeks and 44.1% (95% CI, 38.5-49.7%) for pre-eclampsia with delivery at any gestational age. The current Danish screening strategy using maternal risk factors detected 25.0% of women with pre-eclampsia with delivery < 34 weeks and 19.6% of women with pre-eclampsia with delivery < 37 weeks at a SPR of 3.4%. When applying the FMF algorithm including maternal characteristics, MAP, UtA-PI and PlGF at the fixed SPR of 3.4%, the DRs were 60.5% (95% CI, 36.9-84.1%) for PE with delivery < 34 weeks and 45.2% (95% CI, 32.0-58.5%) for PE with delivery < 37 weeks. CONCLUSION: In this large Danish multicenter study, the FMF algorithm based on maternal characteristics, MAP, UtA-PI, PlGF and PAPP-A predicted 77.4% of cases with pre-eclampsia with delivery < 34 weeks and 66.8% of cases with pre-eclampsia with delivery < 37 weeks of gestation at a SPR of 10%, suggesting that the performance of the algorithm in a Danish cohort matches that in other populations. © 2023 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

Maternal exposure to UV filters: associations with maternal thyroid hormones, IGF-I/IGFBP3 and birth outcomes.

BACKGROUND: Several chemical UV filters/absorbers ('UV filters' hereafter) have endocrine-disrupting properties in vitro and in vivo. Exposure to these chemicals, especially during prenatal development, is of concern. OBJECTIVES: To examine maternal exposure to UV filters, associations with maternal thyroid hormone, with growth factor concentrations as well as to birth outcomes. METHODS: Prospective study of 183 pregnant women with 2nd trimester serum and urine samples available. Maternal concentrations of the chemical UV filters benzophenone-1 (BP-1) and benzophenone-3 (BP-3) in urine and 4-hydroxy-benzophenone (4-HBP) in serum were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The relationships between 2nd trimester maternal concentrations of the three chemical UV filters and maternal serum concentrations of thyroid hormones and growth factors, as well as birth outcomes (weight, height, and head and abdominal circumferences) were examined. RESULTS: Positive associations between maternal serum concentrations of 4-HBP and triiodothyronine (T3), thyroxine (T4), insulin-like growth factor I (IGF-I) and its binding protein IGFBP3 were observed in mothers carrying male fetuses. Male infants of mothers in the middle 4-HBP exposure group had statistically significantly lower weight and shorter head and abdominal circumferences at birth compared to the low exposure group. CONCLUSIONS: Widespread exposure of pregnant women to chemical UV filters and the possible impact on maternal thyroid hormones and growth factors, and on fetal growth, calls for further studies on possible long-term consequences of the exposure to UV filters on fetal development and children's health.

Presence of benzophenones commonly used as UV filters and absorbers in paired maternal and fetal samples.

BACKGROUND: Previous studies have demonstrated widespread exposure of humans to certain benzophenones commonly used as UV filters or UV absorbers; some of which have been demonstrated to have endocrine disrupting abilities. OBJECTIVES: To examine whether benzophenones present in pregnant women pass through the placental barrier to amniotic fluid and further to the fetal blood circulation. METHODS: A prospective study of 200 pregnant women with simultaneously collected paired samples of amniotic fluid and maternal serum and urine. In addition, unique samples of human fetal blood (n=4) obtained during cordocentesis: and cord blood (n=23) obtained at delivery, both with paired maternal samples of serum and urine collected simultaneously, were used. All biological samples were analyzed by TurboFlow-liquid chromatography - tandem mass spectrometry for seven different benzophenones. RESULTS: Benzophenone-1 (BP-1), benzophenone-3 (BP-3), 4-methyl-benzophenone (4-MBP), and 4-hydroxy-benzophenone (4-HBP) were all detectable in amniotic fluid and cord blood samples and except 4-HBP also in fetal blood; albeit at a low frequency. BP-1 and BP-3 were measured at ~10-times lower concentrations in fetal and cord blood compared to maternal serum and 1000-times lower concentration compared to maternal urine levels. Therefore BP-1 and BP-3 were only detectable in the fetal circulation in cases of high maternal exposure indicating some protection by the placental barrier. 4-MBP seems to pass into fetal and cord blood more freely with a median 1:3 ratio between cord blood and maternal serum levels. Only for BP-3, which the women seemed to be most exposed to, did the measured concentrations in maternal urine and serum correlate to concentrations measured in amniotic fluid. Thus, for BP-3, but not for the other tested benzophenones, maternal urinary levels seem to be a valid proxy for fetal exposure. CONCLUSIONS: Detectable levels of several of the investigated benzophenones in human amniotic fluid as well as in fetal and cord blood calls for further investigations of the toxicokinetic and potential endocrine disrupting properties of these compounds in order for better assessment of the risk to the developing fetus.

Description and evaluation of a net energy intake model as a function of dietary chewing index.

Previously, a linear relationship has been found between net energy intake (NEI) and dietary chewing index (CI) of the diet for different types of cattle. Therefore, we propose to generalize and calibrate this relationship into a new model for direct prediction of NEI by dairy cows from CI values (CINE; min/MJ of NE). Furthermore, we studied the forage-to-concentrate substitution rate in this new NEI model. To calibrate the model on a diverse set of situations, we built a database of mean intake from 14 production experiments with a total of 986 primi- and multiparous lactating dairy cows of different breeds fed 136 different diets ad libitum. The NEI were estimated by the Nordic feed evaluation system. The CINE value of diets was estimated from the intake of concentrate, intake of forage neutral detergent fiber (NDFf), particle length of forage, indigestible NDFf/NDFf, body weight, NDFf/body weight, and the content of NE in DM. We show that the slope values in this regression are proportional to the squared intercepts, giving the nonlinear equation NEI=NEI0-k×NEI0a×CINE, where the parameter k represents the decline in NEI with the increasing CINE of the diet and a was estimated to have a value of 2, implying a constant maximum daily chewing time. The intercept NEI0 in the regression of NEI on CINE may be interpreted as metabolic net energy intake capacity of the cows fed without physical constraints on intake. Based on experimental data, the maximum chewing time was estimated as 1/(4 × k). The NEI0 values were parameterized as a linear function of metabolic body size, energy-corrected milk yield (kg/d), days in milk, and days in milk squared. Prediction accuracy was evaluated by mean square prediction error (MSPE) and its decomposition into central tendency, regression, and disturbance, across and within experiments on independent data from 19 experiments including 812 primi- and multiparous lactating dairy cows of different breeds fed 80 different diets ad libitum. The NEI model predicted NEI with an MSPE of 8% of observed, and across the 19 experiments the error central tendency, error regression, and error disturbance were 4.2, 40.6, and 84.9% of MSPE, respectively. The described intake model implies a variable forage-to-concentrate substitution rate as a nonlinear function of NEI0, CINE of forage, and supplementation of concentrate.

Variations in automatically recorded rumination time as explained by variations in intake of dietary fractions and milk production, and between-cow variation.

Individual recording of rumination time (RT) is now possible in commercial dairy herds, through development of a microphone-based sensor, which is able to record RT by the sound of rumination activity. The objectives of this study were to examine the relationship between daily RT and intakes of different dietary fractions, the relationship between RT in minutes per kilogram of dry matter intake (DMI) and milk production, and to examine the variation in RT within and between mid-lactating dairy cows. Data from 3 production trials were used in which a total of 27 different diets were fed. The data contained 761, 290, and 203 daily recordings of RT, milk yield, milk components, DMI, and intake of dietary fractions recorded on 29, 26, and 24 Holstein and Swedish Red cows from trials 1, 2, and 3, respectively. The dietary fractions included forage neutral detergent fiber (NDF), concentrate NDF, crude protein, sugar, starch, and the remaining fraction represented by organic matter--(forage NDF+concentrate NDF+crude protein+sugar+starch). The relationship between the dietary fractions and RT was analyzed in 2 steps. In step 1, the dietary fractions, which were significantly related to RT, were selected and simultaneously checked for multicollinearity between the dietary components; in step 2, a multivariate model, including the effect of repeated measurements, the main effect of the selected dietary fractions from step 1, random effects of cow(trial) and trial, and information on breed, days in milk, and parity was used to analyze the relationship between RT and the selected dietary fractions. Relationships between RT in minutes per kilogram of DMI and milk yield and milk components were analyzed, using the same multivariate model as in step 2. Approximately 32% of the variation in daily RT could be explained by variations in intakes of the dietary fractions, whereas 48% of the total variation in RT was accounted for by individual variations between cows. Intakes of forage NDF and starch were positively related to daily RT, whereas intakes of sugar and the remaining fraction were negatively related to daily RT. Rumination time in minutes per kilogram of DMI was negatively related to milk yield and protein percentage, but positively related to milk fat percentage.

Evaluation of physical structure value in spring-harvested grass/clover silage and hay fed to heifers.

The physical structure value of conserved grass/clover forages of spring harvest was evaluated by assessing effects of harvest time, conservation method, iNDF/NDF ratio and NDF intake (NDFI) per kg BW on chewing activity and fecal particle size in dairy heifers. A mixed sward consisting of ryegrass (Lolium perenne), red clover (Trifolium pratense) and white clover (Trifolium repens) was harvested in 2009 on May 9 (early) and 25 (late), and both cuts were conserved as silage and hay. The early silage, early hay, late silage and late hay contained dry matter (DM) of 454, 842, 250 and 828 g/kg, and NDF of 315, 436, 414 and 503 g/kg DM, respectively. Forages were fed as sole feed to four Jersey heifers of 435±30 kg BW in a 4×4 Latin square experiment. Feeding level was 90% of individual ad libitum intake, divided equally across two daily meals offered at 0800 and 1530 h. Chewing activity was estimated from recorded jaw movements (JM) oscillations continuously logged for 96 h and summarized per 24 h as mean effective rumination time and eating time. Eating behavior was further observed during four 20-min test meals. Weight proportion of large feces particles (>1.0 mm) and geometric mean fecal particle size (GPS) were calculated. Potentially indigestible NDF (iNDF) was estimated by incubation for 288 h in situ. The daily DM intake (DMI) decreased with progressing maturity at harvest (P<0.001) while daily NDFI was unaffected by harvest time (P>0.05). Earlier harvest led to less rumination per kg NDFI (P<0.01), similar eating time per kg NDFI (P>0.05) and similar proportion of large particles (P>0.01) compared with later harvest. Rumination time per kg NDFI decreased with higher NDFI per kg BW (P<0.001) and with lower iNDF/NDF ratio (P<0.01). Content and potential digestibility of NDF was greater in hay than in silage from the same harvest probably due to field loss and therefore confounded effects of conservation method. This study of high digestibility grass/clover silage and hay showed that NDF content and NDFI per kg BW affect fecal particle size and rumination time per kg NDF, and suggests implementation of NDFI per kg BW in systems evaluating physical structure in diets.

Effects of feeding level and NDF content of grass-clover silages on chewing activity, fecal particle size and NDF digestibility in dairy heifers.

The objective of this study was to assess effects of feed intake and NDF content of highly digestible grass-clover silage on chewing behavior, fecal particle size distribution and apparent digestibility in restrictively fed heifers. Four grass-clover silages (Lolium perenne, Trifolium pratense and Trifolium repens) were harvested in 2009 at different regrowth stages, resulting in silages with NDF contents of 312, 360, 371 and 446 g/kg dry matter (DM), respectively. Four rumen-fistulated Jersey heifers (343 ± 32 kg BW) were fed silage at 90% of ad libitum levels in a 4 × 4 Latin square design, replicated with further restricted feeding levels (50%, 60%, 70% or 80% of ad libitum) in a balanced 4 × 4 × 4 Greco-Latin square design. Eating activity was estimated from test meal observations, while rumination activity was estimated from jaw movements logged by a jaw recorder system. Total tract digestibility was estimated from chromic oxide marker and fecal spot sampling, and fecal particle size distribution in washed and freeze-dried particulate DM was determined by dry sieving (2.36, 1.0, 0.5, 0.212 and 0.106 mm, and bottom bowl). Higher NDF content of silage stimulated longer eating time per kg DM intake (P<0.001), while reduced feeding level caused a reduction in eating time per kg DM intake (P<0.001) and NDF (P<0.001). Rumination time per kg DM intake (P<0.01) increased with reduced feeding level, with less effect of feeding level at lower NDF contents (P<0.01) and more rumination with greater NDF content (P<0.01). Relative to NDF intake, rumination time increased with greater NDF content (P<0.01), at a higher rate with reduced feeding level (P<0.05). Digestibility of potentially digestible NDF (DNDF) decreased with greater NDF content (P<0.001) and increased with reduced feeding level (P<0.05). Increasing NDF content resulted in more particulate DM in feces (g/kg DM; P<0.05) and larger mean particle size (P<0.001). In conclusion, feeding heifers with grass-clover silages of decreasing NDF content increased chewing time relative to NDF intake, reduced mean fecal particle size, and increased DNDF digestibility. Restricting feeding level made heifers eat for a shorter time period while rumination and total chewing was increased, causing the ratio between eating and rumination time to decrease with lower intake of forage fiber. Particle size reduction and digestibility depended mostly on changes in NDF content, especially the indigestible NDF content.

Forage fiber effects on particle size reduction, ruminal stratification, and selective retention in heifers fed highly digestible grass/clover silages.

The objective of this study was to investigate the effect of NDF content in highly digestible grass/clover silage on particle size reduction, ruminal stratification, and selective retention in dairy heifers. The reduction in particle size from feed to feces was evaluated and related to feed intake, chewing activity, and apparent digestibility. Four grass/clover harvests (Mixtures of Lolium perenne, Trifolium pratense, and Trifolium repens) were performed from early May to late August at different maturities, at different regrowth stages, and with different clover proportions, resulting in silages with NDF contents of 312, 360, 371, and 446 g/kg DM, respectively, and decreasing NDF digestibility with greater NDF content. Four rumen-fistulated dairy heifers were fed silage at 90% of ad libitum level as the only feed source in a 4 × 4 Latin square design. Silage, ingested feed boluses, medial and ventral ruminal digesta, and feces samples were washed with neutral detergent in nylon bags of 10-µm pore size, freeze dried, and divided into small (<0.212 mm), medium (0.212 to 1 mm), and large : LP; >1 mm) particles by dry-sieving. Chewing activity, rumen pool size, and apparent digestibility were measured. Intake of NDF increased linearly from 2.3 to 2.8 kg/d with greater NDF content of forages (P = 0.01), but silages were exposed to similar eating time (P = 0.55) and rumination time per kg NDF (P = 0.35). No linear effect of NDF content was found on proportion of LP in ingested feed boluses (P = 0.31), medial rumen digesta (P = 0.95), ventral rumen digesta (P = 0.84), and feces (P = 0.09). Greater proportions of DM (P < 0.001) and particulate DM (P = 0.008) were found in medial ruminal digesta compared with ventral rumen, and differences in DM proportion increased with greater NDF content (P = 0.02). Particle size distributions were similar for digesta from the medial and ventral rumen regardless of NDF content of the silages (P > 0.13). The LP proportion was >30% of particles in the ventral and medial rumen, whereas in the feces, the LP proportion was <2%. Particle size stratification of the rumen was undetectable regardless of NDF content of the silages, stressing that the retention mechanism of large undigested particles lies elsewhere than with particle entrapment in the rumen mat. In this study, forage particle breakdown, ruminal stratification, and retention of particles in the rumen were not affected by NDF content of highly digestible grass/clover silages.

Recording rumination time by a rumination monitoring system in Jersey heifers fed grass/clover silage and hay at three feeding levels.

This study was conducted with the objective to validate a rumination monitoring system (RMS). The RMS was developed by SCR Engineers Ltd. (Netanya, Israel) to record individual rumination times in commercial dairy herds. Four heifers were fed 4 forage types at 3 feeding levels in 4 periods in a Latin square design. The forage types were spring growth of grass/clover harvested early (May 9) and late (May 25) conserved as silage and hay, creating 4 forage types: early harvest silage (ES), early harvest hay (EH), late harvest silage (LS), and late harvest hay (LH). Feeding levels were ad libitum, 90% of ad libitum, and 60% of ad libitum. Data were collected for 24 h at ad libitum feeding level, for 96 h at 90% of ad libitum, and for 24 h at 60% of ad libitum. Rumination time was recorded by the RMS in minutes per 2-h interval (RT(RMS2_i)) and per 24-h interval (RT(RMS24)). As a reference method, rumination time was recorded by chewing halters identifying rumination time by jaw movements (JM) and was accumulated into minutes per 2-h interval (RT(JM2_i)) and per 24-h interval (RT(JM24)) over the same time intervals as RT(RMS2_i) and RT(RMS24). Differences between RT(RMS2_i) and RT(JM2_i) were observed for EH, LS, and LH, with differences of 3.4 (P < 0.001), 1.1 (P = 0.03), and 3.3 (P < 0.001) min per 2-h interval. In addition, feeding restrictively at 90% and 60% of ad libitum resulted in greater rumination time recorded by the RMS compared to JM, with differences of 2.6 min per 2-h interval (P < 0.001) for 90% of ad libitum and 1.6 min per 2-h interval (P = 0.01) for 60% of ad libitum. Further, the differences between rumination times recorded by the RMS compared to JM were different among the 4 heifers. Despite these effects, rumination time recorded by the RMS correlated well with rumination time recorded by JM with correlation between RT(RMS2_i) and RT(JM2_i) of r = 0.91 (P < 0.001) and between RT(RMS24) and RT(JM24) of r = 0.79 (P < 0.001).

Particle size alterations of feedstuffs during in situ neutral detergent fiber incubation.

Particle size alterations during neutral detergent fiber (NDF) determination and in situ rumen incubation were analyzed by dry sieving and image analysis to evaluate the in situ procedure for estimation of NDF degradation parameters and indigestible NDF concentration in terms of particle size. Early-cut and late-cut grass silages, corn silage, alfalfa silage, rapeseed meal, and dried distillers grains were examined. Treatments were (1) drying and grinding of forage samples and grinding of concentrates; (2) neutral detergent-soluble (NDS) extraction; (3) machine washing and NDS extraction; (4) 24-h rumen incubation, machine washing, and NDS extraction; and (5) 288-h rumen incubation, machine washing, and NDS extraction. Degradation profiles for potentially degradable NDF were determined and image analysis was used to estimate particle size profiles and thereby the risk for particle loss. Particle dimensions changed during NDF determination and in situ rumen incubation and variations depended on feedstuff and treatment. Corn silage and late-cut grass silage varied most in particle area among feedstuffs, with an increase of 139% between 0 and 24h and a decrease of 77% between 24 and 288 h for corn silage and a decrease of 74% for late-cut grass silage between 24- and 288-h in situ rumen incubation. Especially for late-cut grass silage residues after 288 h in situ rumen incubation, a high mass proportion in the critical zone for escape was found. Particle area decreased linearly with increasing incubation time. Particle loss during in situ rumen incubation cannot be excluded and is likely to vary among feedstuffs.

Effect of harvest time of red and white clover silage on chewing activity and particle size distribution in boli, rumen content and faeces in cows.

The study examined the effects of harvest time of red and white clover silage on eating and ruminating activity and particle size distribution in feed boli, rumen content and faeces in cows. The clover crops were harvested at two stages of growth and ensiled in bales. Red clover crops had 36% and 45% NDF in dry matter (DM) at early (ER) and late (LR) harvest, respectively, and the white clover crops had 19% and 29% NDF in DM at the early (EW) and late (LW) harvest, respectively. The silages were fed restrictively (80% of ad libitum intake) twice daily to four rumen cannulated non-lactating Jersey cows (588 ± 52 kg) in a 4 × 4 Latin square design. Jaw movements (JM) were recorded for 96 h continuously. Swallowed boli, rumen mat, rumen fluid and faeces samples were collected, washed in nylon bags (0.01 mm pore size) and freeze-dried before dry sieving through 4.750, 2.360, 1.000, 0.500, 0.212 and 0.106 mm into seven fractions. The length (PL) and width (PW) values of rumen and faeces particles within each fraction were measured by use of image analysis. The eating activity (min/kg DM intake; P < 0.05) was higher in LR compared with the other treatments. The eating activity (min/kg NDF intake; P < 0.05) was affected by clover type with highest values for white clover silage. The mean ruminating time (min/kg DM), daily ruminating cycles (P < 0.001) and JM during ruminating (P < 0.05) were affected by treatment with increasing values at later harvest time. The proportion of washed particle DM of total DM in boli (P < 0.001), rumen mat (P < 0.001), rumen fluid (P < 0.01) and faeces was (P < 0.001) highest by feeding LR. There were identified two peaks (modes 1 and 2) on the probability density distribution (PDF) of PW values of rumen mat and faeces, but only one peak (mode 1) for PL values. There was no difference in the mean and mode 1 PW and PL value in rumen mat between the four treatments. The mean PL, mode PL, mode 2 PW and mean PW in faeces were highest for LR (P < 0.05). The mean particle size in boli measured by sieving was higher at white clover compared with red clover treatments (P < 0.001) and the highest value in faeces was found in LR (P < 0.01). The two peaks on PDF for width values of rumen mat and faeces particles are most likely related to the leaves and the stems/petioles. In conclusion, the mean total chewing activity per kg DM was lowest for the white clover silage and increased for both silages due to later harvest time. The mean particle size in boli was smallest for LR, whereas the mean PL and PW in faeces were highest for the LR.

Effect of harvest time and physical form of alfalfa silage on chewing time and particle size distribution in boli, rumen content and faeces.

The study examined the effects of physical form and harvest time of alfalfa silage on eating and ruminating activity and particle size distribution in feed boli, rumen content and faeces in dry cows. The alfalfa crop was harvested at two stages of growth (early: NDF 37%, late: NDF 44% in dry matter (DM)), and from each harvest, a chopped (theoretical cutting length: 19 mm) and an unchopped crop was ensiled in bales. The silages were fed restrictively to four rumen cannulated non-lactating Jersey cows (391 ± 26 kg) in a 4 × 4 Latin square design. The cows were fed restrictively 80% of their ad libitum intake twice daily. Chewing activity was recorded for 96 h continuously. Swallowed boli, rumen content, rumen fluid and faeces samples were collected, washed in nylon bags (0.01 mm pore size) and freeze-dried before dry sieving through 4.750, 2.360, 1.000, 0.500 and 0.212 mm pore sizes into six fractions. The length (PL) and width (PW) of particles within each fraction was measured by the use of image analysis. The eating activity (min/kg dry matter intake (P < 0.01) and min/kg NDF (P < 0.05)) was affected by harvest time. The mean ruminating time (min/kg DM) was affected by harvest time (P < 0.01), physical form (P < 0.05) and NDF intake per kg BW (P < 0.01). The proportion of washed particle DM of total DM in boli, rumen content, rumen fluid and faeces was affected by harvest time (P < 0.01) and highest by feeding late-harvested alfalfa silage. Two peaks on the probability density distribution function (PDF) of PW and PL values of boli, rumen content and faeces were identified. Chopping of the silage decreased the mean PL and PW, the most frequent PL (mode) and 95% percentile PL and PW values in boli. In the rumen content, chopping increased the mean PW (P < 0.05). The dimension sizes of faeces particles were not significantly affected by chopping. The mode PW value was lower in rumen content and faeces than in boli (P < 0.001), and the mode PL value was higher in boli and lower in faeces compared with rumen contents (P < 0.001). In conclusion, the mean total chewing activity per kg NDF decreased due to chopping and early harvest time. The mean PL and PW in boli decreased due to chopping and late harvest. The two peak values on the PDF (PL) and PDF (PW) of boli, rumen content and faeces particles are most likely related to the leaf and the stem residues.

Glucocorticoids and histone deacetylase inhibitors cooperate to block the invasiveness of basal-like breast cancer cells through novel mechanisms.

Aggressive cancers often express E-cadherin in cytoplasmic vesicles rather than on the plasma membrane and this may contribute to the invasive phenotype of these tumors. Therapeutic strategies are not currently available that restore the anti-invasive function of E-cadherin in cancers. MDA-MB-231 cells are a frequently used model of invasive triple-negative breast cancer, and these cells express low levels of E-cadherin that is mislocalized to cytoplasmic vesicles. MDA-MB-231 cell lines stably expressing wild-type E-cadherin or E-cadherin fused to glutathione S-transferase or green fluorescent protein were used as experimental systems to probe the mechanisms responsible for cytoplasmic E-cadherin localization in invasive cancers. Although E-cadherin expression partly reduced cell invasion in vitro, E-cadherin was largely localized to the cytoplasm and did not block the invasiveness of the corresponding orthotopic xenograft tumors. Further studies indicated that the glucocorticoid dexamethasone and the highly potent class I histone deacetylase (HDAC) inhibitor largazole cooperated to induce E-cadherin localization to the plasma membrane in triple-negative breast cancers, and to suppress cellular invasion in vitro. Dexamethasone blocked the production of the cleaved form of the CDCP1 (that is, CUB domain-containing protein 1) protein (cCDCP1) previously implicated in the pro-invasive activities of CDCP1 by upregulating the serine protease inhibitor plasminogen activator inhibitor-1. E-cadherin preferentially associated with cCDCP1 compared with the full-length form. In contrast, largazole did not influence CDCP1 cleavage, but increased the association of E-cadherin with γ-catenin. This effect on E-cadherin/γ-catenin complexes was shared with the nonisoform selective HDAC inhibitors trichostatin A (TSA) and vorinostat (suberoylanilide hydroxamic acid, SAHA), although largazole upregulated endogenous E-cadherin levels more strongly than TSA. These results demonstrate that glucocorticoids and HDAC inhibitors, both of which are currently in clinical use, cooperate to suppress the invasiveness of breast cancer cells through novel, complementary mechanisms that converge on E-cadherin.

Temporal effects of maternal and pregnancy characteristics on serum pregnancy-associated plasma protein-A and free ß-human chorionic gonadotropin at 7-14 weeks' gestation.

OBJECTIVE: The aim of this study was to investigate gestational age-dependent effects of racial origin, smoking status and mode of conception on maternal serum levels of free ß-human chorionic gonadotropin (ß-hCG) and pregnancy-associated plasma protein-A (PAPP-A) at 7-14 weeks' gestation. METHODS: This was an analysis of data from prospective first-trimester combined screening for aneuploidies in singleton pregnancies, with ß-hCG and PAPP-A measured at 7 + 1 to 14 + 3 weeks' gestation. We included 27,908 pregnancies from three centers in the U.K. and 125,461 pregnancies from 22 centers in Denmark, all with known normal fetal karyotype or resulting in the birth of a phenotypically normal neonate. Multiple regression modelling of log10 -transformed marker concentrations was used to produce log10 multiple of the median (MoM) values for free ß-hCG and PAPP-A and to examine pregnancy characteristics that have significant effects on marker concentrations. RESULTS: Serum free ß-hCG and PAPP-A concentrations were significantly affected by gestational age, maternal weight, racial origin, parity, smoking and mode of conception. There were significant gestational age-dependent effects attributed to Afro-Caribbean race, smoking and conception through in-vitro fertilization (IVF) on PAPP-A and free ß-hCG levels. In women of Afro-Caribbean race there was a weekly increase in PAPP-A of 5.3% and in free ß-hCG of 1.8%. In smokers there was a weekly decrease in PAPP-A of 2.4% and in free ß-hCG of 1.6%. In cases of IVF conceptions there was a weekly increase in PAPP-A of 4.5% and in free ß-hCG of 4.6%. CONCLUSIONS: Serum free ß-hCG and PAPP-A concentrations at 7-14 weeks' gestation are affected by several pregnancy characteristics. The effects of Afro-Caribbean race, smoking and IVF conception change with gestational age.

Effect of stage of maturity of grass at harvest on intake, chewing activity and distribution of particle size in faeces from pregnant ewes.

This study was conducted to investigate the effect of stage of maturity at harvest on the intake of grass silage, eating and ruminating activity and the distribution of faecal particle size in ewes during late pregnancy. A total of 18 Swedish Finull x Dorset 85 ± 8 kg (mean 6 ± s.d.) ewes bearing twins were randomly assigned to three dietary treatments 6 weeks before lambing. The treatments included ad libitum feeding with early harvested (EH), medium harvested (MH) or late harvested (LH) primary-growth grass silage with 45%, 58% and 63% NDF on a dry matter (DM) basis, respectively. Intake and chewing activity were recorded and faeces were sampled over 4 continuous days for each individual ewe. The faeces samples were washed in nylon bags, freeze dried and sieved with pore sizes from 2.4mm to 0.1 mm; particles less than 0.1mm in size were also collected. Subsamples of each sieving fraction were scanned and the dimensions of the individual particles in each sieving fraction were measured by image analysis. In addition, the number of particles longer than 7 mm was counted from the particles retained on a sieve with a pore size of 2.4mm using a simple wet sieving technique. The time spent eating and ruminating per kg of DM intake was affected by the stage of maturity at harvest; it was shorter in ewes fed EH compared with ewes fed MH and LH ( P < 0.05). In comparison with feeding LH, feeding EH resulted in the retention of a larger proportion of particles in the lower and upper sieve fractions (< 0.2mm and > 1 mm, respectively, P < 0.01), a smaller mean particle size ( P < 0.05) and a smaller mean particle width in faeces ( P < 0.01). The results from the simple wet sieving technique confirmed the results from dry sieving and image analysis, showing a higher number of large particles in faeces from ewes fed the EH compared with the ewes fed the MH and LH (P < 0.001). In conclusion, the distribution of faecal particle size might be considered as a footprint of the characteristics of forage fibre eaten by ewes.

Prospective study evaluating performance of first-trimester combined screening for trisomy 21 using repeat sampling of maternal serum markers PAPP-A and free ß-hCG.

OBJECTIVE: To prospectively evaluate the performance of first-trimester combined screening for trisomy 21 using the biochemical markers pregnancy-associated plasma protein-A (PAPP-A) and free beta-human chorionic gonadotropin (free ß-hCG) obtained before and at the time of the nuchal translucency (NT) scan. METHODS: Three fetal medicine departments in Denmark participated in the study. Screening for trisomy 21 was set up as a two-step approach with blood sampling performed before the NT scan (early sample) and again at the time of the NT scan (late sample). PAPP-A and free ß-hCG were measured on both the early and late samples. Age-standardized detection and false-positive rates for different screening protocols were calculated. RESULTS: We collected two blood samples in 27 pregnancies affected by trisomy 21 and in 3891 control pregnancies. The early samples were taken between gestational ages 8 + 0 and 13 + 6 weeks, and the late samples between 11 + 3 and 14 + 6 weeks. The median interval between the samples was 17 (range, 1-40) days. We found a significantly better estimated screening performance when using early sampling vs late sampling (P < 0.05). With a risk cut-off of 1 in 100, at the time of the risk assessment the estimated detection and false-positive rates when using the early sample were 91% (95% CI, 81-98%) and 1.6% (95% CI, 1.3-2.0%), respectively. For fixed false-positive rates the highest detection rates were achieved using both blood samples. When comparing early sampling vs double sampling there was no significant difference in screening performance. CONCLUSION: In combined first-trimester screening for trisomy 21, use of early sampling with measurement of PAPP-A and free ß-hCG before the time of the NT scan can optimize screening performance. Using maternal serum markers obtained both before and at the time of the NT scan has the potential to further improve performance, but larger studies are needed to confirm this potential.

Early lactation feed intake and milk yield responses of dairy cows offered grass silages harvested at early maturity stages.

The main objective was to evaluate the potential of grass silages of very high quality to support a high milk yield with a low or moderate, or even without concentrate supplementation. Production responses to increased levels of concentrate supplementation with 3 primary growth grass silages differing in digestibility were studied using 66 Norwegian Red dairy cows. Roundbale silage was produced from a timothy-dominated sward at very early (H1), early (H2), and normal (H3) stages of crop maturity. Crops were rapidly wilted (<24h) and a formic acid-based additive was applied. All silages were restrictedly fermented. Silage digestible organic matter in dry matter (DM) values were 747, 708, and 647 g/kg of DM for H1, H2, and H3, respectively. Dietary treatments were fed in a 3×3 factorial arrangement of the 3 silages supplemented with 3 concentrate levels (4, 8, and 12 kg/d) and, additionally, H1 was offered without concentrates and H3 with 16 kg/d, giving a total of 11 diets. Cows, blocked according to parity and calving date, were introduced to the experiment before calving and kept in the experiment until wk 16 of lactation. Silage was offered ad libitum in loose housing and concentrate was available in automatic feed stations. Intake of grass silage when fed as the sole feed was 16.9 kg of DM on average for lactation wk 1 to 16. When H1 was supplemented with 4 or 8 kg of concentrates, silage DM intake did not change, but total DM intake increased to 20.6 and 23.7 kg/d, respectively. Energy-corrected milk (ECM) yield increased from 23.4 kg when H1 was offered without concentrate supplement to 29.1 and 32.8 kg when supplemented with 4 or 8 kg concentrate, respectively. None of the other diets equaled the yield obtained by H1 plus 8 kg of concentrate. Feed intake and yield of cows offered H3 plus 4 kg of concentrates were strongly constrained by high dietary fiber concentration. They consumed 16.5 g of neutral detergent fiber/kg of body weight and spent more time eating silage than cows offered other diets. The highest concentrate level within each silage quality produced similar or lower ECM yield than that with 4 kg less concentrates. The obtained milk yield responses suggest that provision of 8.0, 8.4, and 11.5 kg of concentrates to H1, H2, and H3, respectively, would maximize ECM yield within each silage type. However, H1 may successfully be used with less concentrates, or even without, if future conditions should limit the amount of concentrates available for ruminant production.

Skin temperature measured by infrared thermography after specific ultrasound-guided blocking of the musculocutaneous, radial, ulnar, and median nerves in the upper extremity.

BACKGROUND: Sympathetic block causes vasodilatation and increases in skin temperature (T(s)). However, the T(s) response after specific nerve blocking is unknown. In this study, we hypothesized that T(s) would increase after specific blocking of the nerve innervating that area. METHODS: Forty-six patients undergoing hand surgery were included. We performed ultrasound-guided, specific nerve blocking of either the musculocutaneous, radial, ulnar, or median nerve in each patient and analysed T(s) in the forearm and hand at 2 min intervals in the following 22 min by the use of infrared thermography. Areas of interest corresponding to the cutaneous innervation area of each of the four nerves were defined and the mean T(s) in each area was analysed. RESULTS: Specific blocking of the ulnar and median nerves caused a substantial increase in mean (sd) T(s) in the areas innervated by these nerves [5.2 (3.2)°C and 5.1 (2.5)°C, respectively; both P<0.0001]. The increase was even larger at the fingertips. Median nerve blocking also increased T(s) in the area of the hand innervated by the radial nerve (P<0.0001). However, T(s) did not increase in any area after either musculocutaneous or radial nerve blocking. CONCLUSIONS: Specific blocking of the ulnar and median nerve causes substantial increases in T(s) in specific areas of the hand. In contrast, the specific blocking of the musculocutaneous or radial nerve does not increase T(s). Further studies are needed to clarify if these findings can be used to objectively evaluate brachial plexus block success.

Effects of physical form and stage of maturity at harvest of whole-crop barley silage on intake, chewing activity, diet selection and faecal particle size of dairy steers.

This study examined the effects of physical form and stage of maturity at harvest of whole-crop barley silage (WCBS) on feed intake, eating and rumination activity, diet selection and faecal particle size in dairy steers. Whole-crop barley was harvested and ensiled in round bales. Eight dairy steers (live weight (LW): 350 ± 10 kg) in a duplicated 4 × 4 Latin square design were fed WCBS harvested at heading or dough stage of maturity in long form or chopped in a 2 × 2 factorial treatment arrangement. The WCBS was supplemented with soybean meal. Daily dry matter (DM) intake increased by 7% (P < 0.05) due to chopping of dough-stage silage but was unaffected by chopping of heading-stage silage. The steers fed chopped, but not those fed long dough-stage silage, selected for starch in the WCBS (P < 0.001). The neutral detergent fibre (NDF) intake was 5% higher (P < 0.01) for heading than for dough-stage silage and was associated with lower concentration of indigestible NDF (96 v. 170 g/kg DM). Rate of intake of DM and NDF was 37% higher (P < 0.001), daily eating time was 24% shorter (P < 0.001) and daily chewing time was 8% shorter (P < 0.05) for chopped silage compared with long silage but there was no effect of maturity. Daily rumination time was not affected by treatments, whereas rumination and chewing times per kg NDF intake were 15% and 13% higher (P < 0.05), respectively, for dough-stage than for heading-stage silage. The proportion of faecal particles retained on a 1 mm screen was 30% higher (P < 0.001) due to chopping and 45% higher (P < 0.001) due to delayed harvest. Chopping the dough-stage silage reduced the proportion of grain in faeces from 97 to 43 g/kg DM (P < 0.05) indicating higher starch digestibility. In conclusion, chopping increased DM intake of WCBS when harvested at dough stage but not at heading stage of maturity.