Single-Cell Transcriptomic Analysis of Salivary Gland Endothelial Cells.

Vascular endothelial cells have important tissue-specific functions in fibrosis and regeneration. In the salivary gland, endothelial cells are required for proper development, but their roles within adult glands are largely unknown. To identify ligand-receptor interactions between endothelial cells and other cell types that may be important during fibrosis and regeneration, we used a reversible ductal ligation injury. To induce injury, a clip was applied to the primary ducts for 14 d, and to induce a regenerative response, the clip was subsequently removed for 5 d. To identify endothelial cell-produced factors, we used single-cell RNA sequencing of stromal-enriched cells from adult female submandibular and sublingual salivary glands. Transcriptional profiles of homeostatic salivary gland endothelial cells were compared to endothelial cells of other organs. Salivary gland endothelial cells expressed many unique genes and displayed the highest overlap in gene expression with other fenestrated endothelial cells from the colon, small intestine, and kidney. Comparison of the 14-d ligated, mock-ligated, and 5-d deligated stromal-enriched transcripts and lineage tracing revealed that endothelial cells retain their identity following ligation and recovery from injury. CellChat and NATMI were used to predict changes in ligand-receptor interactions from endothelial cells to other cells in response to ligation and deligation. CellChat and NATMI predicted that after ligation, interactions with fibroblasts, epithelial cells, and glial cells were increased, and following deligation, interactions with pericyte, glia, fibroblasts, and immune cells were increased. Some of the highest-ranked interactions predicted in ligated compared to mock endothelial cells were between glial cells via Col4a2-Cd93 and Jag2-Notch1, as well as epithelial cells via Pecam1-Cd38, while in deligated compared to ligated endothelial cells, the top interactions were between fibroblasts via Ntf3-Ntrk2, glial cells via Hspg2-Itgb1, and pericytes via Jam2-F11r. Understanding salivary gland endothelial cell signaling will inform future endothelial cell-based regenerative therapies.

Assessment of fetal growth trajectory identifies infants at high risk of perinatal mortality.

OBJECTIVE: To analyze perinatal risks associated with three distinct scenarios of fetal growth trajectory in the latter half of pregnancy compared with a referent group. METHODS: This cohort study included women with singleton pregnancies that birthed between 32+0 and 41+6 gestational weeks and had two or more ultrasound scans at least four weeks apart from 18+0 weeks. We evaluated three different scenarios of fetal growth against a referent, defined as appropriate for gestational age-sized fetuses with appropriate forward growth trajectories. The comparator growth trajectories were categorized as Group 1: Small for gestational age (SGA) fetuses (EFW or AC <10th centile) with appropriate forward growth; Group 2: Decreased growth trajectory fetuses (decrease of ≥50 centiles) with EFW or AC ≥10th centile (i.e., non-SGA) at their final scan; and Group 3: Decreased growth trajectory fetuses with EFW or AC <10th centile (i.e., SGA) at their final scan. The primary outcomes were perinatal mortality (stillbirth or neonatal death). Secondary outcomes included stillbirth, birth of an SGA infant, preterm birth, emergency cesarean section (CS) for non-reassuring fetal status (NRFS), and composite severe neonatal morbidity. Associations were analyzed using logistic regression. RESULTS: The final study cohort comprised 5319 pregnancies. Compared to the referent group, the adjusted odds of perinatal mortality were significantly increased in Group 2 (odds ratio [OR] 4.00, 95%CI 1.36-11.22) and Group 3 (OR 7.71, 95%CI 2.39-24.91). Only Group 3 had increased odds of stillbirth (OR 5.69, 95%CI 1.55-20.93). In contrast, infants in Group 1 did not have significantly increased odds of demise. The odds of an SGA infant at birth increased in all three groups but were highest in Group 1 (OR 111.86, 95%CI 62.58-199.95) and Group 3 (OR 40.63, 95%CI 29.01-56.92). In both groups, more than 80% of infants were born SGA, and nearly half had a birth weight <3rd centile. Likewise, the odds of preterm birth were increased in all three groups, being the highest in Group 3 with an OR of 4.27 (3.23-5.64). Lastly, the odds of severe neonatal morbidity were increased in Groups 1 and 3, whereas the odds of emergency CS for NRFS were only increased in Group 3. CONCLUSIONS: Assessing the fetal growth trajectory in the latter half of pregnancy can help identify infants at increased risk of perinatal mortality and birth weight <3rd centile for gestation. This article is protected by copyright. All rights reserved.

Patient preferences in geriatric wards, a survey of health care professionals' practice, experience and attitudes.

PURPOSE: We aimed to identify whether health care professionals (HCP) examine their patient and next-of-kin preferences, and to study whether medical decisions follow these preferences. METHOD: A cross-sectional web-based survey was conducted with multidisciplinary HCP from 12 geriatric wards in the South-Eastern Norway Regional Health Authority. RESULTS: Of the 289 HCPs responding (response rate 61%), mean age 37.8 years (SD 11.3), 235 (81.3%) women, 12.4 (SD 9.6) years of experience and 67 (23.2%) medical doctors, only half report clarifying patients' preferences. The majority reported that they did not inform, involve and treat in line with such preferences. However, 53% believe that HCP, patients and next-of-kin should make clinical decisions together. DISCUSSION: Our findings indicate a lack of engagement in conversation and inclusion of patient preferences when providing health interventions in geriatric wards. Measures for change of culture are needed.

Phenotype analysis of families with TP53 germline variants at the Center for Familial Breast and Ovarian Cancer, Cologne.

PURPOSE: Tumor protein p53 (TP53) pathogenic variant (PV) carriers are identified during genetic testing for hereditary causes of cancer. PVs in TP53 are associated with the Li-Fraumeni syndrome (LFS), and thus, surveillance and preventive measures are important for TP53 PV carriers. However, the penetrance of TP53 PVs can be low if the Chompret criteria are not fulfilled. In this study, we compared the phenotypic characteristics of families that did and did not fulfill the LFS criteria according to Chompret. METHODS: The German Consortium for Hereditary Breast and Ovarian Cancer (GC-HBOC) database was used to identify index patients with a likely pathogenic/pathogenic TP53 variant and their family members. The study investigated the type of variant, pedigree, age of onset, number of primary tumors, and histological type of BC. RESULTS: TP53 PV were present in the index cases of 35 families, 57% (20/35) of which fulfilled the Chompret criteria. The median age of onset at first BC diagnosis was lower in families that fulfilled the Chompret criteria compared to those who did not. Four of all diseased individuals were minors (4%; 4/105) when malignancy was first diagnosed. Sarcomas and brain tumors occurred in 10% (10/105) and in 7% (7/105) of all diseased persons, respectively. BC was the most frequently occurring first tumor (60%; 62/105) and additional malignancy (45%; 20/44) in this cohort. Subsequent malignancies developed in 31% (20/65) of the individuals who fulfilled the Chompret criteria compared with 15% (6/40) of those who did not. CONCLUSION: The tumor spectrum and age of onset found in this study showed that tumors other than BC had low disease penetrance in TP53 PV carriers identified using the GC-HBOC criteria for genetic testing.

Preliminary validity testing of the eHealth Literacy Questionnaire (eHLQ): a Confirmatory Factor Analysis (CFA) in Norwegian hospitalized patients.

AIMS: To perform the first psychometric analysis of the Norwegian version of the eHLQ using confirmative factor analysis (CFA) procedures in a population of patients admitted to hospital using a cross-sectional design. The eHLQ consists of 35 items capturing the 7-dimensional eHealth Literacy Framework (eHLF) which describes users' attributes, user's interaction with technologies and user's experience with digital health systems. METHODS: The 7 independent scales of the eHLQ was translated from Danish and culturally adapted into the Norwegian language following a standardised protocol. Assessment of construct validity of the eHLQ was undertaken using data from a cross-sectional survey of 260 patients hospitalized at a Norwegian University Hospital in the Oslo area during a two-week period in June 2021. The analysis included using correlation analysis (Pearsons R), internal consistency (Cronbach's alpha) and confirmatory factor analysis (CFA). RESULTS: All factor loadings were high to acceptable (i.e. > 0.6), except for five items which had somewhat lower loadings. Regarding internal consistency, alpha ranged from 0.73 to 0.90. For optimal CFA fit for the different scale models, correlated residuals were required for five of the seven scales. Overall our analysis shows an intermediate fit of the orginal construct. Scale intercorrelations were all below 0.8, indicating an overall acceptable discriminant validity between the 7 dimensions. CONCLUSIONS: The results from the CFA analysis indicate that for almost all 7 eHLQ scales, an acceptable model fit was achieved. The 260 hospitalized patients included in this study represented a variety of diagnoses, recruited from a geographically limited area. Further studies on psychometric properties of the Norwegian version of eHLQ in larger samples, diverse settings and by using more comprehensive approaches are warranted.

Ruminal tissue uptake of amino acids in Holstein cows when supply of nutrients within the rumen differs.

Characterisation of amino acid (AA) use by the ruminal vein-drained viscera (RDV) has not been assessed in vivo in dairy cattle, and thus, the extent of ruminal AA use from arterial and postabsorptive blood supplies is unclear. Understanding the complete use of AA by the splanchnic bed may lead to alternative feeding programmes that maximise animal N efficiency. The objective of this work was to determine how different nutritional manipulations affect RDV net appearance and apparent affinity for arterial AA in lactating dairy cattle. Data from two arterio-venous (A-V) difference studies, that used a common set of multicatheterised lactating Holstein cows, assigned to different nutritional treatments, were used to assess ruminal metabolism. Study 1 consisted of three dietary treatments at calving [an alfalfa-glucogenic diet, a glucogenic diet (GLCG), or a ketogenic diet (KETO)] to investigate the effects of dietary nutrients and increasing intake postpartum on RDV metabolism of AA at -14, +4, +15, and +29 days relative to calving (DRTC). Study 2 consisted of two dietary levels of CP (17 or 13%) and three ruminal buffers (ammonia, butyrate, and control) to investigate the level of dietary CP and ruminal fermentation products on RDV metabolism of AA. Blood was collected at 9, 20, and 30 min after buffer administration. Regardless of dietary nutrients or fermentation products present in ruminal fluid, net RDV uptake was positive for most AA, excepting Asp, Cys, Glu, and Ser, which were consistently negative. The general positive net uptakes indicate that any AA potentially absorbed from the rumen were not adequate to meet apparent needs. Ruminal plasma flow and net RDV uptake of Trp, Ala, Gly, and Pro increased linearly with increased DRTC. Feeding KETO or GLCG diets increased ruminal plasma flow, and net RDV uptake of Thr and Gly. Feeding high CP diets increased ruminal uptake of Leu, Phe, and Val. The increased AA uptakes were partially driven by increased plasma flow, however, tissue affinity as reflected in clearance rates also increased or tended to for Met, Trp, Ala, Gly, Pro, and Tyr suggesting that changes in RDV uptake were regulated and not due solely to mass action. In conclusion, splanchnic tissue bed responses to dietary and washed rumen conditions were in part driven by changes in RDV nutrient demand and metabolic activity. The adaptive responses alter the fraction of absorbed AA utilised for non-productive purposes and thus the efficiency of conversion of those AA to product.

Fiber digestibility and protein value of pulp silage for lactating dairy cows: Effects of wet fractionation by screw pressing of perennial ryegrass.

The aim of the study was to investigate the effects of substituting silage of chopped grass with pulp silage of grass fractionated once or twice in a biorefinery using a screw press on fiber kinetics, protein value, and production of CH4 in dairy cows. Six lactating multiparous Holstein cows in mid-lactation (176 ± 93 d in milk), cannulated in the rumen, duodenum, and ileum, were used in an incomplete 6 × 4 Latin square design with a 2 × 3 factorial arrangement of treatments. Perennial ryegrass was harvested in third regrowth from the same field at early and late developmental stage (35 and 44 d of regrowth, respectively) and subjected to 1 of 3 types of processing within each developmental stage. Grass was either harvested for normal silage making (mowed, wilted, chopped, and ensiled), or harvested fresh and fractionated using a screw press. Half of the pulp from the first fractionation was ensiled, whereas the other half of the pulp was rehydrated, fractionated a second time, and pulp hereof was ensiled. The grass and pulp silages were used with concentrates (65:35 forage to concentrate ratio) to make total mixed rations (TMR) based on either silage of chopped grass (GS), pulp silage of grass fractionated once (1×P), or pulp silage of grass fractionated twice (2×P), harvested either at early (E) or late (L) developmental stage resulting in 6 different TMR treatments (EGS, E1×P, E2×P, LGS, L1×P, L2×P). The TMR were fed for ad libitum intake and samples of intestinal digesta and feces were collected for determination of digestibility. The effect of processing on ash-free neutral detergent fiber (aNDFom) concentration in silages depended on developmental stage, but showed that within each developmental stage, pulp silage of grass fractionated twice had higher aNDFom concentration than pulp silage of grass fractionated once and silage of chopped grass. The 2×P resulted in lower (14.9 ± 0.55 vs. 17.5 ± 0.54 kg/d) dry matter intake (DMI) compared with GS. The effects of processing and developmental stage interacted such that apparent total-tract aNDFom digestibility was higher (784 ± 13 vs. 715 ± 13 g/kg) for L2×P compared with LGS, whereas no difference was found between E2×P and EGS. Moreover, the protein value was higher (106 ± 5 vs. 92 ± 5 g AA digested in the small intestine/kg of DMI) for 2×P compared with GS. Unexpectedly, processing had no effect on fractional rate of digestion of digestible aNDFom or CH4 yield (L/kg of DMI), whereas feeding forages harvested at early compared with late developmental stage resulted in lower CH4 yield. Feeding pulp silage of grass fractionated once generally yielded results intermediate to cows fed silage of chopped grass and pulp silage of grass fractionated twice. This study showed that pulp silage of fractionated grass could serve as feed for dairy cows because the fiber digestibility and protein value improved, but further research investigating effects of physical processing of forage on fiber kinetics is required.

Integrated exome and transcriptome analysis prioritizes MAP4K4 de novo frameshift variants in autism spectrum disorder as a novel disease-gene association.

The application of next-generation sequencing (NGS) to clinical practice is still hampered by the ability to interpret the clinical relevance of novel variants and the difficulty of evaluating their effect in specific tissues. Here, we applied integrated genomic approaches for interrogating blood samples of two unrelated individuals with neurodevelopmental disorders and identified a novel neuro-pathogenic role for the Mitogen-Activated Protein Kinase 4 gene (MAP4K4). In particular, we identified two novel frameshift variants in coding exons expressed in the blood and neuronal isoforms. Both variants were predicted to generate non-sense-mediated decay. By transcriptome analysis, we simultaneously demonstrated the deleterious effect of the identified variants on the splicing activity and stability of MAP4K4 mRNA. Therefore, we propose MAP4K4 as a novel causative gene for non-syndromic and syndromic neurodevelopmental disorders. Altogether, we prove the efficacy of an integrated approach of exome and transcriptome sequencing in the resolution of undiagnosed cases by leveraging the analysis of variants in genes expressed in peripheral blood.

Effects of changes in diet energy density and milking frequency and a single injection of cabergoline at dry-off on feeding behavior and rumination time in dairy cows.

Dry-off is a typical management practice, but research on the effects of dry-off on feeding behavior in high-yielding cows is limited. The present study investigated the effects of 2 diet energy densities: lactation diet (normal energy density, NORM) versus a lactation diet diluted with 30% straw (reduced energy density, REDU), both offered ad libitum, and 2 daily milking frequencies (2× vs. 1×) during the 7 d before dry-off day (d 0), and the effects of an injection of either a dopamine agonist [cabergoline (CAB); Velactis, Ceva Santé Animale; labeled for use only with abrupt dry-off; i.e., no reduction in feeding level or milking frequency before the last milking] or saline (SAL) following the last milking on d 0 (2 × 2 × 2 factorial arrangement), on automatically monitored feeding behavior and rumination time in 119 clinically healthy, loose-housed, pregnant, lactating Holstein cows during the week before and after d 0. From d 0, all cows were fed the same dry-cow diet ad libitum. Data were analyzed in R using mixed-effects models. Over the days before d 0, REDU cows spent 30% more time feeding at a 50% lower feeding rate, visited both assigned and unassigned feed bins more frequently, and spent more time ruminating than NORM cows. No clear behavioral effects of reduced milking frequency were found. Within 24 h following injection, CAB cows spent approximately 40% less time feeding at a lower feeding rate, visited their feed bin 28% less often, and spent 40% less time ruminating than SAL cows, irrespective of treatment before dry-off. The current study demonstrates that reducing diet energy density for 1 wk before dry-off led to clear behavioral changes in high-yielding cows. Administering CAB after the last milking induced decreased feeding behavior lasting approximately 24 h, indicating collateral effects other than reduced prolactin secretion.

Feeding up to 91% concentrate to Holstein and Jersey dairy cows: Effects on enteric methane emission, rumen fermentation and bacterial community, digestibility, production, and feeding behavior.

Due to climate change, periods of drought might be longer and occur more frequently, which challenges roughage production and requires changed feeding of dairy cattle by increasing the grain content of the diet. This study investigated the effect of diets with concentrate proportions up to 91% of dry matter on dry matter intake (DMI), milk production, enteric methane emission, rumen fermentation, rumen bacterial community structure, nutrient digestibility, and feeding behavior of Holstein and Jersey dairy cows. Twelve Danish Holstein and 12 Danish Jersey cows were fed ad libitum with one of 3 total mixed rations differing in concentrate proportion in a continuous design with staggered approach over 19 to 29 d. Dietary concentrate proportions were 49% (C49), 70% (C70), and 91% (C91) on dry matter basis, and were based on increasing proportions of chopped barley straw, dried beet pulp, barley, NaOH-treated wheat, dried distillers grain, and rapeseed cake at the expense of grass/clover silage, corn silage and soybean meal. Cows were adapted to the diets over a 12- to 19-d period, before rumination activity was measured over 3 d. Subsequently, spot samples of feces were collected for digestibility determination over 2 d, and gas exchange was measured on the last 3 d of the experimental period. Shortly after chamber stay, rumen liquid was collected using an oro-ruminal device. Dry matter intake was higher for Holstein than Jersey. Methane emissions (all expressions) were affected by the interaction between breed and diet. Methane per kilogram of DMI was lowered by 18 and 48% for Holstein fed C70 and C91, respectively, compared with C49, whereas this was 17 and 22% respectively for Jersey. Rumen propionate molar proportion increased more, rumen bacterial community was less diverse, and rumination time and rumination chews relative to DMI reduced less for Holstein than for Jersey cows with increasing concentrate level. In conclusion, Holstein dairy cows responded stronger to increased dietary concentrate level regarding methane mitigation, changes in rumen VFA profile, and effect on the rumen bacterial community structure than Jersey cows, whereas Jersey cows responded stronger with regard to rumination time and rumination chews (per kilogram of DMI and per kilogram of neutral detergent fiber intake) than Holstein cows. Thus, diets high in concentrates are a less effective methane mitigation strategy for Jersey than for Holstein.

Effects of particle size and toasting of fava beans and forage source on nutrient digestibility, ruminal fermentation, and metabolizable protein supply in dairy cows.

The objective of this study was to investigate the effects of heat treatment (toasting) and particle size alterations (grinding; rolling) on nutrient utilization, ruminal fermentation, and supply of metabolizable protein (MP), and to study the interaction between processing conditions of fava beans and forage type. Six Danish Holstein dairy cows fitted with ruminal, duodenal, and ileal cannulas were used in a 6 × 4 incomplete Latin square design with 4 periods of 21-d duration. Cows were fed ad libitum with 6 experimental diets: diets high in either grass-clover silage or corn silage were combined with ground untoasted, ground toasted, or rolled untoasted fava beans. Samples of ruminal fluid, digesta from duodenum and ileum, and feces were collected, and nutrient digestibility was estimated using Cr2O3 and TiO2 as flow markers. Diets high in corn silage resulted in higher ruminal pH and higher proportion of propionate in ruminal volatile fatty acids compared with diets high in grass-clover silage. Diets high in corn silage resulted in higher apparent total-tract digestibility of crude protein and starch but lower apparent ruminal and total-tract digestibility of neutral detergent fiber compared with diets high in grass-clover silage. Rolling of fava beans decreased the in situ small intestinal disappearance of rumen-undegradable protein corrected for particle losses. Compared with grinding, rolling of fava beans reduced apparent ruminal digestibility of starch, true ruminal digestibility of organic matter, crude protein, and AA, and small intestinal digestibility of AA and starch. Grinding of fava beans increased apparent ruminal digestibility of neutral detergent fiber and reduced the proportion of propionate in ruminal volatile fatty acids compared with rolling of fava beans. In addition, rolling of fava beans had no effect on MP supply. Toasting of fava beans had no effect on in vivo nutrient digestibility except for an interaction with forage source on apparent ruminal dry matter and organic matter digestibility. Toasting of fava beans did not affect small intestinal digestion of individual and total AA, and therefore failed to increase MP supply. In conclusion, neither replacing grass-clover silage with corn silage, nor toasting nor rolling of fava beans had an effect on supply of MP.

Dual Sounding Rocket and C/NOFS Satellite Observations of DC Electric Fields and Plasma Density in the Equatorial E- and F-Region Ionosphere at Sunset.

E × B plasma drifts and plasma number density were measured on two NASA rockets launched simultaneously at sunset from Kwajalein Atoll with apogees of 182 and 331 km, with similar, coincident measurements gathered on the Communications/Navigation Outage Forecasting System (C/NOFS) satellite at 390 km. The combined measurements portray a highly dynamic ionosphere in a narrow range of local time and altitude, providing evidence of vortex-like motions. Although the vertical plasma drift was upwards, its magnitude was not constant, increasing between ∼150 and 250 km altitude where the plasma density was reduced. The zonal plasma drifts displayed a shear with altitude, changing from eastward to westward flow below 270 km, coincident with the larger upward drifts and consistent with the maintenance of the vortex flow. The plasma density on the western flank was highly structured compared to the eastern flank, despite the fact that the western region corresponded to slightly earlier local times. These observations illustrate that the low latitude ionosphere at sunset must be considered as an ensemble of interconnected flows encompassing an evolving "theater," as opposed to a background that simply unfolds linearly with respect to local time. The observations also underscore how satellites at high altitudes do not capture the highly dynamic ionosphere and thermosphere at the lower altitudes which are critical for understanding the electrodynamics system. Such motions set the stage for large scale plasma instabilities to form later in the evening, as observed by radars at Kwajalein and subsequent passes of the C/NOFS satellite.

Nitrogen isotopic discrimination as a biomarker of between-cow variation in the efficiency of nitrogen utilization for milk production: A meta-analysis.

Estimating the efficiency of N utilization for milk production (MNE) of individual cows at a large scale is difficult, particularly because of the cost of measuring feed intake. Nitrogen isotopic discrimination (Δ15N) between the animal (milk, plasma, or tissues) and its diet has been proposed as a biomarker of the efficiency of N utilization in a range of production systems and ruminant species. The aim of this study was to assess the ability of Δ15N to predict the between-animal variability in MNE in dairy cows using an extensive database. For this, 20 independent experiments conducted as either changeover (n = 14) or continuous (n = 6) trials were available and comprised an initial data set of 1,300 observations. Between-animal variability was defined as the variation observed among cows sharing the same contemporary group (CG; individuals from the same experimental site, sampling period, and dietary treatment). Milk N efficiency was calculated as the ratio between mean milk N (grams of N in milk per day) and mean N intake (grams of N intake per day) obtained from each sampling period, which lasted 9.0 ± 9.9 d (mean ± SD). Samples of milk (n = 604) or plasma (n = 696) and feeds (74 dietary treatments) were analyzed for natural 15N abundance (δ15N), and then the N isotopic discrimination between the animal and the dietary treatment was calculated (Δ15n = δ15Nanimal - δ15Ndiet). Data were analyzed through mixed-effect regression models considering the experiment, sampling period, and dietary treatment as random effects. In addition, repeatability estimates were calculated for each experiment to test the hypothesis of improved predictions when MNE and Δ15N measurements errors were lower. The considerable protein mobilization in early lactation artificially increased both MNE and Δ15N, leading to a positive rather than negative relationship, and this limited the implementation of this biomarker in early lactating cows. When the experimental errors of Δ15N and MNE decreased in a particular experiment (i.e., higher repeatability values), we observed a greater ability of Δ15N to predict MNE at the individual level. The predominant negative and significant correlation between Δ15N and MNE in mid- and late lactation demonstrated that on average Δ15N reflects MNE variations both across dietary treatments and between animals. The root mean squared prediction error as a percentage of average observed value was 6.8%, indicating that the model only allowed differentiation between 2 cows in terms of MNE within a CG if they differed by at least 0.112 g/g of MNE (95% confidence level), and this could represent a limitation in predicting MNE at the individual level. However, the one-way ANOVA performed to test the ability of Δ15N to differentiate within-CG the top 25% from the lowest 25% individuals in terms of MNE was significant, indicating that it is possible to distinguish extreme animals in terms of MNE from their N isotopic signature, which could be useful to group animals for precision feeding.

Standardised or individualised X-ray tube angle for mediolateral oblique projection in digital mammography?

INTRODUCTION: We aimed to investigate whether there were any differences in positioning criteria related to the presentation of the pectoralis major muscle (pectoral muscle) for women of different heights using a standardized 60° X-ray tube angle for mammograms in mediolateral oblique (MLO) projection. METHODS: Data from MLO mammograms of right breasts of 45,193 women screened in BreastScreen Norway 2016-2019 were used. The positioning criteria were related to the pectoral muscle length (measure A and measure B), width and shape and considered adequate or inadequate depending on the degree of fulfilling the criteria. Data associated with the pectoral muscle were extracted from Volpara, an automated software for breast density assessment. Information on height was obtained from a self-reported questionnaire received by the women together with the invitation to attend the screening program. Women were divided into three groups based on the height percentiles (P) in the Norwegian growth curves: < 25th percentile (75th percentile (>P75th: >170 cm). Logistic regression was used to analyse the odds of adequate pectoral muscle length A and B, and shape, adjusting each model for screening technique and equipment model. Results were presented with odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Mean age of the screened women was 61.5 (SD = 4.8) years. The adequate measure for the pectoral muscle length A was obtained for 25.9% (11,724/45,193), length B for 76.3% (34,489/45,193), width for 75.0% (33,894/45,193) and shape for 97.6% (44,118/45,193) of the mammograms. Adjusted odds of an adequate pectoral muscle length A were lower for women of P75th (OR = 1.08, 95% CI 1.02-1.14) compared to women of P25-75. Odds of an adequate pectoral muscle shape were higher for women of P75th (OR = 0.92, 95% CI 0.87-0.97) compared to women of P25-75th. CONCLUSION: The 60° X-ray tube angle might suit most of the female population offered mammographic screening in Norway, but women of a relatively low height (163 cm or lower) might benefit from an X-ray tube angle less than 60-degrees. IMPLICATIONS FOR PRACTICE: Using 60° X-ray tube angle for the MLO mammograms in BreastScreen Norway fit the majority of the participating women. More research is needed to change the protocol associated with the tube angle for women shorter than 163 cm.

Effects of dietary and milking frequency changes and administration of cabergoline on clinical udder characteristics in dairy cows during dry-off.

We investigated the effects of 2 diet energy densities [normal lactation diet (NORM) vs. energy-reduced diet (REDU), both fed for ad libitum intake] and 2 daily milking frequencies [twice (2×) vs. once (1×)] during 1 week before the dry-off day, as well as effects of an injection of either a dopamine agonist [cabergoline (CAB); Velactis, Ceva Santé Animale, Libourne, France; labelled for use only with abrupt dry-off, e.g. no reduction in diet energy density or milking frequency before the last milking] or saline (SAL) following the last milking, on clinical udder characteristics of Holstein cows. During a week before and after the last milking, the following measures were recorded: palpation-based udder firmness and soreness; image-based hock-hock distance; responsiveness to mechanical udder stimulation and degree of udder fill measured with a dynamometer. Before the last milking, REDU cows displayed lower odds of having a firm udder and lower degree of udder fill, as well as lower responsiveness to mechanical udder stimulation, than NORM cows. After the last milking, REDU cows displayed shorter hock-hock distance compared with NORM cows. The effects of milking frequency on the clinical udder characteristics were unclear. Within 24 h following injection, CAB cows showed lower odds of having a firm udder, shorter hock-hock distance, and lower degree of udder fill than SAL cows, irrespective of treatment group before dry-off. In this study, reducing diet energy density prior to dry-off, and to some extent administering the dopamine agonist cabergoline after the last milking, resulted in fewest clinical udder changes.

The use of advanced platelet-rich fibrin after surgical removal of mandibular third molars: a systematic review and meta-analysis.

The objective of this systematic review was to test the hypothesis of no difference in recovery following surgical removal of mandibular third molars with the application of advanced platelet-rich fibrin (A-PRF) in the extraction socket compared with alternative biomaterials or natural wound healing. A search of MEDLINE (PubMed), Embase, Cochrane Library, and Scopus was conducted. Human randomized controlled trials published in English up until December 31, 2020 were included. Outcome measures were pain, facial swelling, trismus, soft tissue healing, alveolar osteitis, and quality of life; these were evaluated by descriptive statistics and meta-analysis including 95% confidence intervals (CI). Four studies with a low or moderate risk of bias fulfilled the inclusion criteria. A-PRF resulted in significantly lower pain scores when compared with leucocyte platelet-rich fibrin or natural wound healing after 2 days (-16.8, 95% CI -18.9 to -14.7), 3 days (-12.1, 95% CI -13.4 to -10.7), and 7 days (-1.9, 95% CI -2.9 to -0.9). A-PRF seems to have a negligible effect on facial swelling and trismus and some beneficial effect on soft tissue healing. Alveolar osteitis and quality of life were not assessed. The included studies were characterized by considerable heterogeneity and confounding variables. Thus, the level of evidence appears to be inadequate for clinical recommendations according to the focused question.

The role of QT-prolonging medications in a forensic autopsy study from Western Denmark.

Medication-induced prolongation of the QT-interval (miQTP) can lead to cardiac arrhythmia. Our aim was to investigate the prevalence of forensic autopsy cases where fatal cardiac arrhythmia related to treatment with QT-prolonging medications (QT-PMs) could be suspected. We performed a cross-sectional study of 741 forensic autopsies undertaken at our institution in non-drug addicts aged 15 years or above from 2017 to 2019. We defined a high risk of miQTP by one detected QT-PM in a concentration above therapeutic level, or two or more detected QT-PMs in post mortem blood. We reviewed the autopsy reports from cases with a high miQTP-risk to identify cases with no other apparent cause of death. We discarded suicides and cases with lethal levels of QT-PMs. We identified 167 cases (22.5%) with high risk of miQTP, and discarded 36 suicides (4.9%) and 7 (0.9%) with lethal levels of QT-PMs. Apart from a high risk of miQTP, no other apparent explanation of the cause of death was present in seven (0.9%). In 18 cases (2.4%) with high miQTP-risk, the cause of death was primarily attributed to cardiac changes other than acute cardiovascular events. In conclusion, 22.5% had a high risk of miQTP, and fatal cardiac arrhythmia related to treatment with QT-PMs could be suspected in 0.9%. However, a genetic pro-arrhythmic background could not be excluded in our study. Furthermore, it is possible that QT-PMs could have played a role in some of the 2.4% of cases where the cause of death was mainly attributed to cardiac changes and the risk of miQTP was high.

Clinically decisive (dis)agreement in multidisciplinary team assessment of esophageal squamous cell carcinoma; a prospective, national, multicenter study.

BACKGROUND: Decisions regarding tumor staging, operability, resectability, and treatment strategy in patients with esophageal cancer are made at multidisciplinary team (MDT) conferences. We aimed to assess interobserver agreement from four national MDT conferences and whether this would have a clinical impact. METHODS: A total of 20 patients with esophageal cancer were included across all four upper gastrointestinal (GI) cancer centers. Fully anonymized patient data were distributed among the MDT conferences which decided on TNM category, resectability, operability, curability, and treatment strategy blinded to each other's decisions. The interobserver agreement was expressed as both the raw observer agreement and with Krippendorff's α values. Finally, a case-by-case evaluation was performed to determine if disagreement would have had a clinical impact. RESULTS: A total of 80 MDT evaluations were available for analysis. A moderate to near-perfect observer agreement of 79.2%, 55.8%, and 82.5% for TNM category was observed, respectively. Substantial agreement for resectability and moderate agreement for curability were found. However, an only fair agreement was observed for the operability category. The treatment strategies had a slight agreement which corresponded to disagreement having a clinical impact in 12 patients. CONCLUSIONS: Esophageal cancer MDT conferences had an acceptable interobserver agreement on resectability and TM categories; however, the operability assessment had a high level of disagreement. Consequently, the agreement on treatment strategy was reduced with a potential clinical impact. In future MDT conferences, emphasis should be on prioritizing the relevant information being readily available (operability, T & M categories) to minimize the risk of disagreement in the assessments and treatment strategies, and thus, delayed or suboptimal treatment.

Daily alternation of the dietary starch level in Holstein dairy cows.

The aim of this study was to investigate the effect of controlled daily alternations in dietary starch level on changes in rumen environment, blood, urine, and milk metabolites of dairy cows. Six multiparous mid-lactation Holstein cows were used in a replicated 3 × 3 Latin square design with 14-d periods and 3 alternating levels of dietary starch as treatments. Each 14-d period consisted of a 7-d baseline period and 7-d alternating period where diets alternated day to day. During the baseline period, all cows were fed a control diet containing 21% starch (dry matter basis). During the alternating period, the control diet was replaced with 1 of the 3 experimental diets on d 8, 10, 12, and 14. The 3 experimental diets contained 28% (low), 35% (medium), and 42% (high) starch (dry matter basis). At d 7 (baseline), 8 (ALT1), and 14 (ALT4) of each period, rumen fluid, blood, urine, and quarter milk (i.e., back right quarter) samples were collected at -0.5, 1, 2.5, 4, 5.5, and 7 h relative to morning feeding (0800 h). No differences were observed in dry matter intake, milk yield, and milk chemical composition. Rumen medial pH was lower in the high alternation level compared with the low or medium alternation levels at ALT1 but did not differ among starch alternation levels at ALT4. Similarly, the difference between rumen pH in medial and ventral contents was reduced at ALT1 with high alternation level but was not affected at ALT4. Total volatile fatty acid (VFA) concentrations were higher in the rumen medial fluid of the high alternation level at 7 h relative to morning feeding compared with those from the low and medium alternation levels. Similarly, total VFA concentrations constantly increased and were the highest in the ventral rumen fluid at 7 h relative to morning feeding, although no differences were detected among starch alternation levels. In both rumen medial and ventral fluids, the high alternation level showed higher propionate and lower acetate proportions compared with low and medium alternation levels. No differences in blood pH were detected among starch alternation levels. However, glucose concentrations tended to be higher in cows from the high alternation level. l-Lactate concentrations in blood were higher in ALT1 than in ALT4 but were not affected by the starch alternation level. In urine, no differences in pH or l-lactate concentrations were detected among alternation levels (i.e., low, medium, and high). Similarly, no differences in milk pH were detected among alternation levels. According to these results, it seems that the daily dietary starch alternation from 21% up to 42% (dry matter basis) is able to affect the ruminal fluid, especially during the first alternation. However, these changes in rumen fluid did not cause any effect on the variables measured in blood, urine, or milk. This study indicates that cows can cope with day-to-day alternations in type of rumen fermentable organic matter; however, longer-term effects on performance and health should be addressed in future studies.

Assessing effects of dietary and milking frequency changes and injection of cabergoline during dry-off on hunger in dairy cows using 2 feed-thwarting tests.

We investigated the single and combined effects of 2 feeding levels (normal lactation diet vs. energy-reduced diet, both fed for ad libitum intake) and 2 daily milking frequencies (twice vs. once) during 1 wk before the dry-off day (d 0), as well as an intramuscular injection of either a dopamine agonist (cabergoline; Velactis, Ceva Santé Animale; labeled for use only with abrupt dry-off, e.g., no reduction in feeding level or milking frequency before the last milking) or saline after the last milking on d 0 on the feeding motivation of clinically healthy, loose-housed, pregnant, lactating Holstein cows. From d 0, all cows were fed the same dry-cow diet for ad libitum intake. Cows were subjected to 2 feed-thwarting tests, a test in the home pen using their diets (test A: d -6, -1, and 1; during 35 min when the feed bins were filled, but locked) and another test carried out in an adjacent pen in which access to concentrate provided in a familiar plastic box was blocked by a wire-mesh lid (test B: d -5 and 2). In test A, we recorded how often cows attempted to feed per 35 min, whether cows vocalized during the 35-min period, and latency to feed within 300 s after feed bins were unlocked. In test B, we recorded latency to approach either of 2 familiar boxes (the wire-mesh box and an identical open box with a small portion of concentrate) within 600 s and how often cows directed behaviors toward the wire-mesh box (number of occurences/5 min). On d -6 (test A), no clear differences in feeding motivation among treatments were found. On d -5 and -1, cows fed the energy-reduced diet displayed a higher probability of vocalizing (test A), were more than 50% quicker to feed (test A), were approximately 5× quicker to approach a box (test B), and directed 60% more behavior toward the wire-mesh box (test B) than cows fed the normal diet. Moreover, cows fed the energy-reduced diet attempted to feed approximately 75% more on d -1 compared with d -6 (test A). On d 2 (test B), cows previously fed the normal diet directed 40% more behavior toward the wire-mesh box than cows previously fed the energy-reduced diet. Reducing feeding level, either before or on the dry-off day, resulted in consistently increased feeding motivation, interpreted as a sign of hunger. No clear effects of change in milking frequency, singly or combined with reduced diet energy density, on feeding motivation were found before d 0. Whereas, on d 2, cows previously milked twice daily were quicker to approach a box than cows previously milked once daily. Cows injected with cabergoline attempted to feed more, but showed lower probability of vocalizing compared with saline-injected cows (d 1; test A), irrespective of treatment before d 0. The effects of cabergoline on feeding motivation are not easily interpreted and warrant further investigation. From a hunger perspective, reducing milking frequency rather than diet energy density seems to be a less negative management to reduce milk production before dry-off.