Studying the Longest ‘Legal’ U.S. Same-Sex Couples: A Case of Lessons Learned

We review methodological opportunities and lessons learned in conducting a longitudinal, prospective study of same-sex couples with civil unions, recruited from a population-based sample, who were compared with same-sex couples in their friendship circle who did not have civil unions, and heterosexual married siblings and their spouse. At Time 1 (2002), Vermont was the only US state to provide legal recognition similar to marriage to same-sex couples; couples came from other US states and other countries to obtain a civil union. At Time 2 (2005), only one US state had legalized same-sex marriage, and at Time 3 (2013) about half of US states had legalized same-sex marriage, some within weeks of the onset of the Time 3 study. Opportunities included sampling legalized same-sex relationships from a population; the use of heterosexual married couples and same-sex couples not in legalized relationships as comparison samples from within the same social network; comparisons between sexual minority and heterosexual women and men with and without children; improvements in statistical methods for non-independence of data and missing data; and the use of mixed methodologies. Lessons learned included obtaining funding, locating participants over time as technologies changed, and on-going shifts in marriage laws during the study

Organoids: a promising new in vitro platform in livestock and veterinary research.

Organoids are self-organizing, self-renewing three-dimensional cellular structures that resemble organs in structure and function. They can be derived from adult stem cells, embryonic stem cells, or induced pluripotent stem cells. They contain most of the relevant cell types with a topology and cell-to-cell interactions resembling that of the in vivo tissue. The widespread and increasing adoption of organoid-based technologies in human biomedical research is testament to their enormous potential in basic, translational- and applied-research. In a similar fashion there appear to be ample possibilities for research applications of organoids from livestock and companion animals. Furthermore, organoids as in vitro models offer a great possibility to reduce the use of experimental animals. Here, we provide an overview of studies on organoids in livestock and companion animal species, with focus on the methods developed for organoids from a variety of tissues/organs from various animal species and on the applications in veterinary research. Current limitations, and ongoing research to address these limitations, are discussed. Further, we elaborate on a number of fields of research in animal nutrition, host-microbe interactions, animal breeding and genomics, and animal biotechnology, in which organoids may have great potential as an in vitro research tool

Continuous real-time cow identification by reading ear tags from live-stream video

In precision dairy farming there is a need for continuous and real-time availability of data on cows and systems. Data collection using sensors is becoming more common and it can be difficult to connect sensor measurements to the identification of the individual cow that was measured. Cows can be identified by RFID tags, but ear tags with identification numbers are more widely used. Here we describe a system that makes the ear tag identification of the cow continuously available from a live-stream video so that this information can be added to other data streams that are collected in real-time. An ear tag reading model was implemented by retraining and existing model, and tested for accuracy of reading the digits on cows ear tag images obtained from two dairy farms. The ear tag reading model was then combined with a video set up in a milking robot on a dairy farm, where the identification by the milking robot was considered ground-truth. The system is reporting ear tag numbers obtained from live-stream video in real-time. Retraining a model using a small set of 750 images of ear tags increased the digit level accuracy to 87% in the test set. This compares to 80% accuracy obtained with the starting model trained on images of house numbers only. The ear tag numbers reported by real-time analysis of live-stream video identified the right cow 93% of the time. Precision and sensitivity were lower, with 65% and 41%, respectively, meaning that 41% of all cow visits to the milking robot were detected with the correct cow’s ear tag number. Further improvement in sensitivity needs to be investigated but when ear tag numbers are reported they are correct 93% of the time which is a promising starting point for future system improvements

Early locomotor activity in broilers and the relationship with body weight gain

Fast-growing broilers are relatively inactive and this is thought to be a result of selection for high growth rates. This reduced activity level is considered a major cause of leg weakness and associated leg health problems. Increased activity, especially early in life, is suggested to have positive effects on leg health, but the relationship between early activity and growth is unclear. A clearer understanding of the relationship between activity early in life and body weight gain could help determine how selecting on increased early activity could affect body weight gain in broilers. Here, a radio frequency identification (RFID) tracking system was implemented to record daily individual broiler activity throughout life, in 5 production rounds. As mean activity levels alone do not capture the variation in activity over time, multiple (dynamic) descriptors of activity were determined based on the individual birds’ daily distances moved, focusing on the period from 0 to 15 days old. The mean, skewness, root mean square error (RMSE), autocorrelation, and entropy of (deviations in) activity were determined at the individual level, as well as the average daily gain (ADG). Relationships between activity descriptors and ADG were determined for 318 birds. Both when combining the data from the different production rounds and when taking production round and start weight into account, a negative relationship between ADG and RMSE was observed, indicating that birds that were more variable in their activity levels had a lower ADG. However, the activity descriptors, in combination with recording round and start weight, explained only a small part (8%) of the variation in ADG. Therefore, it is recommended for future research to also record other factors affecting ADG (e.g., type of feed provided and feed intake) and to model growth curves. Overall, this study suggests that increasing early activity does not necessarily negatively affect body weight gain. This could contribute to improved broiler health and welfare if selecting for increased activity has the expected positive effects on leg health

Genetic parameters for social effects on survival in cannibalistic layers: Combining survival analysis and a linear animal model

<p>Abstract</p> <p>Background</p> <p>Mortality due to cannibalism in laying hens is a difficult trait to improve genetically, because censoring is high (animals still alive at the end of the testing period) and it may depend on both the individual itself and the behaviour of its group members, so-called associative effects (social interactions). To analyse survival data, survival analysis can be used. However, it is not possible to include associative effects in the current software for survival analysis. A solution could be to combine survival analysis and a linear animal model including associative effects. This paper presents a two-step approach (2STEP), combining survival analysis and a linear animal model including associative effects (LAM).</p> <p>Methods</p> <p>Data of three purebred White Leghorn layer lines from Institut de Sélection Animale B.V., a Hendrix Genetics company, were used in this study. For the statistical analysis, survival data on 16,780 hens kept in four-bird cages with intact beaks were used. Genetic parameters for direct and associative effects on survival time were estimated using 2STEP. Cross validation was used to compare 2STEP with LAM. LAM was applied directly to estimate genetic parameters for social effects on observed survival days.</p> <p>Results</p> <p>Using 2STEP, total heritable variance, including both direct and associative genetic effects, expressed as the proportion of phenotypic variance, ranged from 32% to 64%. These results were substantially larger than when using LAM. However, cross validation showed that 2STEP gave approximately the same survival curves and rank correlations as LAM. Furthermore, cross validation showed that selection based on both direct and associative genetic effects, using either 2STEP or LAM, gave the best prediction of survival time.</p> <p>Conclusion</p> <p>It can be concluded that 2STEP can be used to estimate genetic parameters for direct and associative effects on survival time in laying hens. Using 2STEP increased the heritable variance in survival time. Cross validation showed that social genetic effects contribute to a large difference in survival days between two extreme groups. Genetic selection targeting both direct and associative effects is expected to reduce mortality due to cannibalism in laying hens.</p

Whole Slide Imaging Versus Microscopy for Primary Diagnosis in Surgical Pathology: A Multicenter Blinded Randomized Noninferiority Study of 1992 Cases (Pivotal Study)

Most prior studies of primary diagnosis in surgical pathology using whole slide imaging (WSI) versus microscopy have focused on specific organ systems or included relatively few cases. The objective of this study was to demonstrate that WSI is noninferior to microscopy for primary diagnosis in surgical pathology. A blinded randomized noninferiority study was conducted across the entire range of surgical pathology cases (biopsies and resections, including hematoxylin and eosin, immunohistochemistry, and special stains) from 4 institutions using the original sign-out diagnosis (baseline diagnosis) as the reference standard. Cases were scanned, converted to WSI and randomized. Sixteen pathologists interpreted cases by microscopy or WSI, followed by a wash-out period of ≥4 weeks, after which cases were read by the same observers using the other modality. Major discordances were identified by an adjudication panel, and the differences between major discordance rates for both microscopy (against the reference standard) and WSI (against the reference standard) were calculated. A total of 1992 cases were included, resulting in 15,925 reads. The major discordance rate with the reference standard diagnosis was 4.9% for WSI and 4.6% for microscopy. The difference between major discordance rates for microscopy and WSI was 0.4% (95% confidence interval, -0.30% to 1.01%). The difference in major discordance rates for WSI and microscopy was highest in endocrine pathology (1.8%), neoplastic kidney pathology (1.5%), urinary bladder pathology (1.3%), and gynecologic pathology (1.2%). Detailed analysis of these cases revealed no instances where interpretation by WSI was consistently inaccurate compared with microscopy for multiple observers. We conclude that WSI is noninferior to microscopy for primary diagnosis in surgical pathology, including biopsies and resections stained with hematoxylin and eosin, immunohistochemistry and special stains. This conclusion is valid across a wide variety of organ systems and specimen types

Feasibility and effectiveness of trifluridine/tipiracil in metastatic colorectal cancer: real-life data from The Netherlands

Background: The RECOURSE trial showed clinical efficacy for trifluridine/tipiracil for refractory metastatic colorectal cancer patients. We assessed the feasibility and effectiveness of trifluridine/tipiracil in daily clinical practice in The Netherlands. Methods: Medical records of patients from 17 centers treated in the trifluridine/tipiracil compassionate use program were reviewed and checked for RECOURSE eligibility criteria. Baseline characteristics, safety, and survival times were compared, and prespecified baseline characteristics were tested in multivariate analyses for prognostic significance on overall survival (OS). Results: A total of 136 patients with a median age of 62 years were analyzed. Forty-three patients (32%) did not meet the RECOURSE eligibility criteria for not having received all prior standard treatments (n = 35, 26%) and/or ECOG performance status (PS) 2 (n = 12, 9%). The most common grade ≥3 toxicities were neutropenia (n = 44, 32%), leukopenia (n = 8, 6%), anemia (n = 7, 5%), and fatigue (n = 7, 5%). Median progression-free survival (PFS) and median OS were 2.1 (95% CI, 1.8–2.3) and 5.4 months (95% CI, 4.0–6.9), respectively. Patients with ECOG PS 2 had a worse median OS (3.2 months) compared to patients with ECOG PS 0–1 (5.9 months). ECOG PS, KRAS-mutation status, white blood cell count, serum lactate dehydrogenase, and alkaline phosphatase were prognostic factors for OS. Conclusions: Our data show that treatment with trifluridine/tipiracil in daily clinical practice is feasible and safe. Differences in patient characteristics between our population and the RECOURSE study population should be taken into account in the interpretation of survival data. Our results argue against the use of trifluridine/tipiracil in patients with ECOG PS 2. Funding: Johannes J.M. Kwakman received an unrestricted research grant from Servier

DNA Methylation Profiles of Ovarian Clear Cell Carcinoma

BACKGROUND: Ovarian clear cell carcinoma (OCCC) is a rare ovarian cancer histotype that tends to be resistant to standard platinum-based chemotherapeutics. We sought to better understand the role of DNA methylation in clinical and biological subclassification of OCCC. METHODS: We interrogated genome-wide methylation using DNA from fresh frozen tumors from 271 cases, applied non-smooth non-negative matrix factorization (nsNMF) clustering, and evaluated clinical associations and biological pathways. RESULTS: Two approximately equally sized clusters that associated with several clinical features were identified. Compared to Cluster 2 (N=137), Cluster 1 cases (N=134) presented at a more advanced stage, were less likely to be of Asian ancestry, and tended to have poorer outcomes including macroscopic residual disease following primary debulking surgery (p-values <0.10). Subset analyses of targeted tumor sequencing and immunohistochemical data revealed that Cluster 1 tumors showed TP53 mutation and abnormal p53 expression, and Cluster 2 tumors showed aneuploidy and ARID1A/PIK3CA mutation (p-values <0.05). Cluster-defining CpGs included 1,388 CpGs residing within 200 bp of the transcription start sites of 977 genes; 38% of these genes (N=369 genes) were differentially expressed across cluster in transcriptomic subset analysis (p-values <10(−4)). Differentially expressed genes were enriched for six immune-related pathways, including interferon alpha and gamma responses (p-values < 10(−6)). CONCLUSIONS: DNA methylation clusters in OCCC correlate with disease features and gene expression patterns among immune pathways. IMPACT: This work serves as a foundation for integrative analyses that better understand the complex biology of OCCC in an effort to improve potential for development of targeted therapeutics