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Modern dog and cat owners increasingly use internet resources to obtain information on pet health issues. While access to online information can improve owners' knowledge of patient care and inform conversations with their veterinarian during consultations, there is also a risk that owners will misinterpret online information or gain a false impression of current standards in veterinary medicine. This in turn can cause problems or tensions, for example if the owner delays consulting their veterinarian about necessary treatment, or questions the veterinarian's medical advice. Based on an online questionnaire aimed at dog and cat owners in Austria, Denmark and the United Kingdom ( N  = 2117) we investigated the use of internet resources to find veterinary medical information, the type of internet resources that were used, and whether owner beliefs explain how often they used the internet to find medical information about their pet. Approximately one in three owners reported that they never used internet resources prior to (31.7%) or after (37.0%) a consultation with their veterinarian. However, when owners do make use of the internet, our results show that they were more likely to use it before than after the consultation. The most common internet resources used by owners were practice websites (35.0%), veterinary association websites (24.0%), or 'other' websites providing veterinary information (55.2%). Owners who believe that the use of internet resources enables them to have a more informed discussion with their veterinarians more often use internet resources prior to a consultation, whereas owners who believed that internet resources help them to make the right decision for their animal more often use internet resources after a consultation. The results suggest that veterinarians should actively ask pet owners if they use internet resources, and what resources they use, in order to facilitate open discussion about information obtained from the internet. Given that more than a third of pet owners use practice websites, the findings also suggest that veterinarians should actively curate their own websites where they can post information that they consider accurate and trustworthy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Springer, Lund, Corr and Sandøe.)

Modern veterinary medicine offers a level of care to cats and dogs similar to that available to their owners, including blood transfusions, chemotherapy and MRI scans. The potential benefits to the animals of owners who can afford such care are obvious, but there can also be negative consequences if owners with strong emotional attachments to their pets pursue treatments that significantly reduce the quality of the animal's life while attempting to prolong it. Moreover, caring for a chronically or seriously ill animal can lead to emotional distress and financial and practical challenges for the pet owner. A questionnaire was used to survey cat and dog owners from representative samples of citizens in the UK, Austria and Denmark, to investigate owners' expectations and attitudes towards advanced veterinary care, and the factors that might influence those views. Overall, 58.4% of the pet owners surveyed believed that their pets should have access to the same treatment options as humans, while 51.5% believed that they should have access to the same diagnostic tests as humans. Owners were most likely to be neutral on the question of whether advanced veterinary care has 'gone too far' (45.3%), and to disagree with the statement that advanced care is 'unnecessary' (40.1%). In all three countries, the level of attachment owners had to their pets was most strongly associated with attitudes towards advanced care, with owners scoring higher on Lexington Attachment to Pets Scale (LAPS) being more likely to expect advanced care to be available. Other factors such as owner age, living situation (alone or not), income or possession of pet insurance were less consistently with owner attitudes. Our findings will help inform veterinarians and other health care providers about pet owner expectations and attitudes towards advanced veterinary care, and contribute to the debate on increasing specialisation within the profession., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Corr et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Telemedicine offers benefits to clients and their animals, but potential disadvantages are also being debated., Methods: Using a questionnaire, we investigated dog and cat owners' (N = 2117) use of and beliefs about telemedicine and whether beliefs impact past and expected future use., Results: Although the majority of owners are aware that telemedicine can lead to the risk of something being missed, they see great potential in remote consultation in terms of usefulness for follow-up appointments or improving access to a specialist. However, only 12% of dog owners and 6% of cat owners have used telemedicine, and around 25% of owners who have never used it would be willing to use it in the future. Owners with a larger number of recent veterinary visits were more likely to have used telemedicine., Limitations: Although a definition of 'telemedicine' was provided, respondents may have had different perceptions of what this meant., Conclusion: Owners of dogs and cats recognise the potential benefits of telemedicine, but there is a mismatch with the actual uptake. This not only raises questions about the current availability of telemedicine but also should increase veterinary professionals' understanding of its potential benefits in veterinary practice., (© 2023 The Authors. Veterinary Record published by John Wiley & Sons Ltd on behalf of British Veterinary Association.)

Previous studies have shown that cat owners seem to care less about their cats than dog owners care about their dogs - both in terms of their emotional state of attachment and in their willingness to pay for services that potentially benefit the animals. One study speculated that this difference is "driven by the behavior of the pet" - that the behavior of dogs encourages care more than the behavior of cats - and therefore is a universal phenomenon. However, previous studies mostly relied on convenience sampling of owners and were undertaken in single countries. Based on responses to a questionnaire from cat and dog owners drawn from representative samples of citizens (18 to 89 years of age) in three different European countries, Denmark, Austria and the United Kingdom, we tested the degree to which owners care about their cats and dogs. We used four different measures: Lexington attachment to pets scale (LAPS), possession of pet health insurance, willingness to pay for life-saving treatment, and expectation of veterinary diagnostic and treatment options. Dog owners had higher LAPS scores in all countries. However, the difference between dog and cat owners was greater in Denmark than in Austria and the United Kingdom. More dogs than cats were insured in all three countries, but the ratio was much less skewed in favor of dogs in the United Kingdom compared to Denmark. In terms of expensive life-saving treatment, in every country, more dog owners than cat owners were willing to spend over a certain amount, but the differences were much more pronounced in Denmark compared to the United Kingdom. In Denmark and Austria, dog owners expected more veterinary treatment options to be available, but species made no difference to the expectations of UK owners. People care more about their dogs than their cats in all countries, but with a clear cross-country variation and a very modest difference in the United Kingdom. Therefore, it does not seem to be a universal phenomenon that people care much less about their cats than their dogs. This finding has practical implications for future efforts to expand the level of veterinary services provided for cat owners., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Sandøe, Palmer, Corr, Springer and Lund.)

Concerns over compromised companion rabbit ( Oryctolagus cuniculus domesticus ) welfare are widespread. The welfare problems have been linked to the perception of rabbits as low investment 'children's pets.' To test this hypothesis and investigate the current conditions for rabbits, data were gathered from two surveys in 2021: a nationally representative survey of Danish companion animal owners (Survey I) and a detailed social media-based survey of Danish rabbit owners (Survey II). Using logistic regression, three owner-related variables (whether a child/adult was responsible for care of the rabbit, owner-opinion on rabbits' suitability as 'starter pets' and willingness-to-pay [WTP] for veterinary treatment) were employed to investigate the effect of rabbit status on owner-provision of selected husbandry conditions. The 76 (Survey I) and 4,335 (Survey II) responses suggested that most rabbits are acquired for children and are solitarily housed, and that many are kept in cages of an unsuitable size and not checked daily. Owners who perceived rabbits as 'starter pets' and with lower WTP were more likely to house rabbits in restricted space and to not provide continuous gnawing opportunities, ad libitum hay or routine healthcare. A child fulfilling the role of the rabbit's main caretaker was also associated with inadequate housing type and fewer gnawing opportunities. Thus, many rabbits live in unsuitable conditions, and owners who perceive rabbits as low investment 'children's pets' are more likely to not provide recommended resources. Changing owners' perceptions of rabbits and promoting suitable husbandry through official education programmes and minimum requirements is important if there are to be improvements made to rabbit welfare., Competing Interests: One author (CRS) was a board member and consulting on rabbit welfare for Kaninværnet (Danish NGO for companion rabbit welfare) in 2021. However, apart from sharing the survey online and providing insights into the companion rabbit sector, the organisation had no impact on the study design, data collection, analyses or interpretation, nor on the manuscript. The remaining authors have no competing interests to declare., (© The Author(s) 2023.)

Reducing one's consumption of foods containing animal products, or avoiding such foods altogether, has become part of everyday life for many people in the Western world. People's motivations for such "animal product limiting" are well-established, but the ways in which individuals enact and experience dietary change in the initial phase are not well understood. Nor is it clear whether, and how, these people present their dietary changes to others. Through the analysis of interviews with 28 people residing in Denmark who had recently (<9 months) embarked on flexitarian, pescetarian, vegetarian or vegan diets, this paper explores how people, in the initial phase of trying to consume fewer, or no, foods with animal products, (i) engage in the practicalities of daily food activities and (ii) communicate their experiences with, and opinions about, the dietary changes they are making in interpersonal interactions. The findings reveal two very different ways of organising the daily food activities: Foodism and Convenience. They also disclose three different ways of communicating in interpersonal interactions: Ethical advocacy, Plant food demonstration and Anonymisation of diet. The paper offers insights into the variation in practices underlying animal product limiting. It suggests that the plant food sector should cater for people relying on convenient food practices as well as those engaged in more advanced ("foodie") practices. Further, in discussing interpersonal communication in the light of community-based social marketing, we argue that the findings highlight how animal product limiters, in everyday social life, may be able to encourage more people to embark on animal product limiting., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to declare., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Exploring everyday life dynamics in meat reduction - a cluster analysis of flexitarians in Denmark. Flexitarians are attracting increasing attention in the research on meat reduction. But there has been limited focus on comprehensive understandings of a broader range of dynamics that can work as barriers and facilitators for meat reduction. In this article, we use social practice theory (SPT) as a comprehensive approach to barriers and facilitators in meat reduction in everyday life. We present an analysis of data from a representative Danish cross-sectional survey. We show, first, that Danish flexitarians can be divided into four distinct clusters (what we will refer to as classes) in accordance with combinations of everyday facilitators and barriers. Second, we show that the prevalence of these classes varies considerably depending on how long people have been flexitarians. We argue that the patterns in this variation indicate that over time people transition to other classes where barriers to plant-rich eating become less significant, and routinization emerges in different ways. Finally, third, we show that flexitarians do report eating less meat than consumers who label themselves as eating meat with no restrictions. But we also highlight that the difference is relatively modest. Indeed, meat intake is still quite common even in classes where routinization is highest. Throughout the paper, we discuss similarities and differences between the SPT framework and another recent framework, the COM-B model, that also provides a comprehensive approach to the understanding of behavioural change., Competing Interests: Declaration of competing interest There are no conflict of interests related to this manuscript., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Assisted reproductive technologies and genetic technologies can accelerate progress in breeding programs in dairy farming, but it is unclear how consumers will react to the use of these technologies. Using representative questionnaire data on Danish citizens (n = 2,036) this cross-sectional study examined consumer attitudes to the application of advanced technologies in dairy cattle breeding. Attitudes were examined in 2 ways. First, we prompted about general attitudes to assisted reproductive technologies and genetic technologies in dairy cow breeding. Here we found that most of the participants were critical of cow impregnation involving hormone therapy and the insertion of cloned fetuses. Second, we used a vignette experiment to study whether acceptance of and willingness to drink milk varies with the type of technique that farmers use for their breeding work, as well as the traits being bred for. We included 5 breeding methods with differing degrees of technological complexity. Participants were randomly assigned to receive tailored information about 1 of the 5 breeding methods. The information specified that dairy farmers' own use of advanced technologies is limited to using semen in artificial insemination on the farm. The potentially concerning technologies are here not applied at farm level but are represented in the semen used in artificial insemination because they were used by breeders on earlier generations of cows and bulls to develop semen with higher genetic merit. There was much less concern about this indirect use of the technologies. Only 1 in 5 participants thought the most advanced method we prompted about (use of semen from breeding methods involving genetic engineering and cloning) was unacceptable. Unwillingness to drink milk from cows produced through such a breeding method was also modest (18%) and not much higher than the unwillingness to drink milk from a cow produced by natural fertilization (10%). A likely reason for the unexpectedly low level of unwillingness to drink milk is that people regard the genetic engineering as distant from the final product. We also found that high-frequency organic milk consumers were more critical of advanced breeding methods. Thus, 28% within this group were unwilling to drink milk from cows impregnated with semen derived from earlier generations of cows and bulls bred using gene editing and cloning. Further, this share rose if the high-frequency organic consumers were very averse to the manipulation of nature. The organic sector may need to cater to this subgroup (e.g., by ensuring the traceability of the semen that organic farmers use to artificially inseminate their cows)., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Aim: The aim of this study was to explore how two of the main dimensions of internalized weight stigma (IWS), namely weight-related self-devaluation and distress, play out in the lives of people with excess weight (EW), and to study whether there are further dimensions of IWS., Method: Ten informants with EW were included in this study. The sample size was determined prior to data collection based on available resources at the time of data collection. All informants both participated in in-depth interviews and responded to the questionnaire WBIS-2F consisting of the two subscales: weight-related self-devaluation and distress. The interview accounts were thematically coded and compared with the informants' scoring on WBIS-2F., Findings: Seven themes were identified from the in-depth interviews: (1) devaluation of competencies, (2) self-blame, (3) bodily devaluation, (4) ambivalence, (5) anticipated stigma, (6) coping strategies, and (7) mental well-being. Overall, the informants scored low on the WBIS-2F subscale weight-related self-devaluation and high on the subscale weight-related distress. The qualitative findings echo the informants' scoring on WBIS-2F. However, novel aspects of IWS not covered by WBIS-2F were also identified. In particular, bodily devaluation presented itself as an integral part of IWS., Conclusion: The two current dimensions of WBIS-2F were retrieved, but important additional aspects of IWS were also identified. Future research is needed to evaluate and test both qualitatively and quantitatively whether the additional aspects of IWS identified in this exploratory examination are separate constructs of IWS., Competing Interests: ED and MP are currently employed at Steno Diabetes Center Copenhagen, a public hospital and research institution in the Capital Region of Denmark which is funded partly by a grant from the Novo Nordisk Foundation. PS regularly collaborates with employees of Novo Nordisk on subjects other than those covered in this paper. PS and TL have previously received a grant from Novo Nordisk to study the attitudes of GPs toward people with overweight., (Copyright © 2023 Davidsen, Pico, Sandøe and Lund.)

Questionable authorship practices in scientific publishing are detrimental to research quality and management. The existing literature dealing with the prevalence, and perceptions, of such practices has focused on the medical sciences, and on experienced researchers. In contrast, this study investigated how younger researchers (PhD students) from across the faculties view fair authorship attribution, their experience with granting guest authorships to more powerful researchers and their reasons for doing so. Data for the study were collected in a survey of European PhD students. The final dataset included 1,336 participants from five European countries (Denmark, Hungary, Ireland, Portugal, and Switzerland) representing all major disciplines. Approximately three in ten reported that they had granted at least one guest authorship to "a person in power". Half of these indicated that they had done so because they had been told to do so by the person in power. Participants from the medical, natural and technical sciences were much more likely to state that they had granted a guest authorship than those from other faculties. We identified four general views about what is sufficient for co-authorship. There were two dominant views. The first (inclusive view) considered a broad range of contributions to merit co-authorship. The second (strongly writing-oriented) emphasised that co-authors must have written a piece of the manuscript text. The inclusive view dominated in the natural, technical, and medical sciences. Participants from other faculties were more evenly distributed between the inclusive and writing oriented view. Those with an inclusive view were most likely to indicate that they have granted a guest authorship. According to the experiences of our participants, questionable authorship practices are prevalent among early-career researchers, and they appear to be reinforced through a combination of coercive power relations and dominant norms in some research cultures, particularly in the natural, technical, and medical sciences., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Goddiksen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Veterinary medicine is increasingly affected by animal owners having the opportunity to become better informed on pet health issues by using various internet resources. Using an online questionnaire including a section on clients' use of internet resources to obtain medical information, this study aimed to investigate veterinarians' estimates of the percentage of clients using internet resources, how often clients question veterinarians' professional medical advice based on online information, and veterinarians' attitudes towards clients' use of internet resources, across Austrian, Danish, and UK veterinarians ( n = 641). The results show that 48.8% of respondents estimated that 40-79% of their clients use internet resources to find medical information. Further, 70-80% of respondents stated that they are occasionally challenged by clients questioning their advice based on online information. Although veterinarians recognized the potential advantages related to clients' use of internet resources, such as an increased acceptance of advanced diagnostics and treatments, they also highlighted clients' increased expectations or false impressions of small animal practices as potentially negative aspects in this context. As internet use increases, it seems likely that these issues will become increasingly important in the future.

Background: Health insurance offers many benefits to clients and veterinarians, such as the ability to perform necessary and possibly cost-intensive medical interventions without financial constraints, or to potentially prevent euthanasia based on financial challenges. However, concerns about negative consequences, such as the overuse of diagnostic tests or overtreatment, have also been raised., Methods: Using an online questionnaire distributed via e-mail, which included a section on health insurance, we investigated the relative number of insured dogs and cats treated by Austrian, Danish and UK veterinarians (N = 636) and the attitudes of those veterinarians toward health insurance. Further, using a case vignette, we examined whether coverage by health insurance may influence treatment suggestions., Results: Even though veterinarians in all three countries believe that health insurance reduces stress since clients' financial resources will be less important, we found that Austrian veterinarians are more likely to agree that health insurance is unnecessary compared to Danish and UK veterinarians. Further, many raised the concern that insurance policies influence clinical decisions; and less than half supported the idea of making insurance mandatory. A majority of veterinarians in Austria and the UK thought that insurance can lead to the overuse of diagnostic tests, and in the UK a majority also thought that it can lead to overtreatment. Using case vignettes, we found that veterinarians were significantly more likely to suggest a CT scan to a client with an insured animal, in contrast to a client with stated financial limitations. Further, UK veterinarians were more likely to suggest a CT scan to a client with an insured animal, in contrast to a client without insurance., Conclusion: In conclusion, we found that veterinarians, in general, were in favour of health insurance, and that greater coverage may increase more cost-intensive veterinary care. Our findings also raise a potential ethical challenge of health insurance causing differential access to clinical care for patients., (© 2022 The Authors. Veterinary Record published by John Wiley & Sons Ltd on behalf of British Veterinary Association.)

Background: Social media is increasingly used in small animal practice, enabling veterinarians to connect with clients and promote their business online. It can also be used by clients to quickly distribute complaints via online communities., Material/methods: Using a questionnaire study we investigated Austrian, Danish and UK veterinarians' attitudes towards Facebook, the contents of clients' online complaints and how they were handled by veterinarians ( N  = 648)., Results: In Denmark and the UK, around 90% of practices had a Facebook page, in contrast to 40% of Austrian practices. Most Danish and UK veterinarians agreed that the use of Facebook was relevant and expected by clients. Agreement was lower among Austrian veterinarians, probably reflecting the lower uptake of social media there. In particular, younger veterinarians and those who actively used Facebook for the practice, could see benefits. In all three countries, we found that clients most frequently complained about treatment costs. Most veterinarians preferred to actively deal with clients' complaints, either replying online or discussing them directly., Conclusions: We recommend future research focusing on veterinarians' personal use of social media and on clients' use of and attitudes towards social media in the veterinary context., Competing Interests: The authors declare they have no conflicts of interest., (© 2022 The Authors. Veterinary Record Open published by John Wiley & Sons Ltd on behalf of British Veterinary Association.)

This cross-country study investigates the potential to improve pig welfare by exploiting consumer demand, domestically and in export markets, for welfare pork produced in indoor production systems. The analysis is based on questionnaire data collected in 2019 focusing on demand for Danish welfare pork both in Denmark and in two nearby export markets, Sweden and Germany. To reduce hypothetical bias, a willingness-to-pay indicator is combined with an indicator of positive interest in buying a fictive Danish welfare labelled pork. We find that the market potential is relatively weak. Our findings indicate that there is some, albeit limited, potential in Denmark and Germany while demand is practically non-existing in Sweden, probably because the pig welfare guaranteed by Swedish legislation is similar to what is provided by the fictive welfare label employed in the study. Hence, consumer demand alone cannot secure enhanced pig welfare. Moreover, we found national differences in the characteristics of consumers who are interested in Danish welfare pork., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

A novel technology combining in vitro production and genomic embryo selection is currently under development in dairy cattle breeding. Adoption of this technology will probably accelerate genetic progress toward the main breeding goals of economic interest, as well as allow selection for traits of societal concern such as decreased methane emissions and improved animal welfare. However, dairy farmers, and especially organic farmers, could find the technology morally questionable and reject its use. This cross-sectional study surveyed Danish dairy farmers' general acceptance of the combined technology and their reported likelihood of using semen produced with it. Drawing on diffusion theory, a questionnaire was developed to examine the way farmers discover and communicate about new technological breeding options, and to measure the factors which predict acceptance and likelihood of adopting the technology. The questionnaire was sent to a randomly selected sample of organic and conventional dairy farmers in Denmark, and 85 organic and 71 conventional farmers (41% response rate) completed it. Seventy-six percent of farmers reported that they would be likely to use semen from bulls derived from the technology. A majority (61%) also found the technology acceptable, but many (33%) were unsure or undecided. Most farmers saw the technology as beneficial, but ethical reservations were aired by around a fifth of the farmers. There were no differences between organic and conventional farmers in likelihood of using, perceived utility, and ethical reservations about the technology. Self-reported idealistic organic farmers showed lower acceptance of the technology, but reported similar likelihood of using semen produced by it. Young farmers (20-39 yr) exhibited higher acceptance of the technology. Larger producers (in terms of number of cows) were more likely to report that they will use and accept the technology. We conclude that it is likely that semen from the technology combining in vitro production and genomic selection would be widely used by both organic and conventional farmers provided that costs can be kept low, and that there are advantages in terms of achieving breeding goals. Structural developments, growth in size of dairy farms, acceptance by young farmers, and the fact that economic incentives (and even ethical arguments) seem to favor the technology all point to this conclusion., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

Small animal veterinarians frequently have to manage conflicting interests. Beside the key consideration of the patient's interests, small animal veterinarians are often challenged to consider not only client's emotional needs, but also their own personal aspirations to provide quality patient care and to make a good living as a professional. Further, veterinarians have an interest in continuous professional development and the use of the newest treatments, which may influence their decision-making process. Based on published work, we hypothesize the existence of four decision ethics orientations that veterinarians can use to manage potentially conflicting concerns. These are: the patient-focused, the client-empathetic, the client-devolved and the development-oriented decision ethics orientations. We surveyed small animal veterinarians in Austria, Denmark, and the UK using a questionnaire (N = 648), and successfully identified the four decision ethics orientations in all three countries. The patient-focused and client-empathetic decision ethics orientations are salient in all countries, whereas Danish and UK veterinarians are slightly more client-empathetic and client-devolved compared to their Austrian colleagues. Across countries our findings show that experienced and older veterinarians tend to be more client-empathetic. Younger and less experienced professionals are more development-oriented compared to their older and more experienced colleagues. In contrast to other studies investigating ethical issues in small animal practice, we found no evidence that gender plays a decisive role in the tendency towards any decision ethics orientation. We also show that veterinarians with a higher client-empathetic orientation and development-orientation more often discuss the possibility of health insurance with clients who do not have it. The present study provides a first empirical insight into how veterinarians manage challenging expectations and ethical concerns as part of decision making in modern small animal practice., Competing Interests: The authors have declared that no competing interests exist.

Background: The relationship between animal ethics orientations and consumer demand for meat with high standards of animal welfare, and the way this relationship plays out in different countries, is not well understood. Using pork as a case study, this comparative study aims to identify the animal ethics orientations that drive purchases of welfare meat in Denmark, Germany, and Sweden., Methods: Cross-sectional questionnaire data from representative samples of approximately 1600 consumers in each country were collected. A segmentation of pork consumers (using latent profile analysis) was carried out., Results: In all three countries, two subgroups were concerned about farm animal welfare: the first subgroup was driven by animal rights values; the second subgroup by animal protection values, where the main principle was that "it is all right to use animals as long as they are treated well". Other consumer groups are less concerned about farm animal welfare and display little or no preference for welfare pork., Conclusions: In all three countries, dual demand for welfare pork exists. The findings of this study can be used, among others, to understand the marketability of enhanced welfare animal products and the potential for market-driven animal welfare improvements.

Background: Bariatric surgery leads to a substantial weight loss (WL), however, a subset of patients undergoing surgery fails to achieve adequate WL. The reason for the individual variation in WL remains unexplained. Using an exploratory cross-disciplinary approach, we aimed to identify preoperative and early postoperative factors explaining the variation in WL after bariatric surgery., Methods: Sixty-one subjects were recruited. Eighteen subjects did not receive surgery and three subjects dropped out, leaving a total sample of 40 subjects. Physiological, social, and psychological data were collected before and 6 months after surgery. All variables were analyzed in combination using a least absolute shrinkage and selection operator (LASSO) regression to explain the variation in WL 18 months after Roux-en-Y gastric bypass (n = 30) and sleeve gastrectomy (n = 10)., Results: Mean WL was 31% (range: 10-52%). The following preoperative factors predicted 59% of the variation in WL: type of surgery (14%), diabetes status (12%), economic resources (9%), sex (7%), binge eating disorder (7%), degree of depression (5%), household type (3%), and physical activity (1%). Including information on early responses after surgery increased the ability to predict WL to 78% and was explained by early WL (47%), changes in energy density of food consumed from a buffet meal (9%), changes in glicentin (5%), degree of depression (5%), sex (5%), type of surgery (2%), economic resources (2%), and changes in drive for thinness (1%)., Conclusions: Using a cross-disciplinary approach, a substantial part of the individual variation in WL was explained by a combination of basic patient characteristics, psychological profile, and social conditions as well as physiological, psychological and behavioral responses to surgery. These results suggest that patient characteristics collected in a cross-disciplinary approach may help determine predictors for less successful WL after bariatric surgery. If verified in larger cohorts this may form the basis for individualized postoperative support to optimize WL outcome.

Knowledge of risk factors for canine obesity is an important pre-requisite of effective preventative strategies. This study aimed to investigate risk factors for canine obesity in adult companion dogs across Zealand, Denmark. Client-owned dogs (>2 years of age and without chronic illness) were recruited and examined at eight companion animal veterinary practices in areas with varying socio-economic characteristics. The body condition score (BCS) of the dogs was examined by two investigators based on a 9-point scoring scheme. Dog owners answered a questionnaire that had prompts regarding: 1) dog characteristics, including neuter status, 2) owner characteristics, 3) feeding and exercise practices and 4) the owners' attachment to the dog. The effect of these factors on BCS and the risk of being heavy/obese (BCS scores 7-9) were analysed in two separate analyses. A total of 268 dogs were included in the analysis, of which 20.5% were found to be heavy/obese. The average BCS was 5.46. In terms of dog characteristics, neutering dramatically increased both BCS and the risk of being heavy/obese in male dogs but not in bitches. BCS and the risk of being heavy/obese increased in senior bitches and decreased in senior male dogs. The risk of being heavy/obese was higher in dogs with overweight and obese owners. Regarding feeding and exercise practices, providing only one meal per day increased BCS and risk of being heavy/obese. Treats during relaxation increased the risk of dogs being heavy/obese. It also increased the dogs' BCS, but only if the owners were overweight or obese. An increased duration of daily walking increased the risk of the dog being heavy/obese, but only if the owner was overweight or obese. Allowing the dog to run free in the garden/property decreased the risk of the dog being heavy/obese. The owners' attachment to the dog was not associated with the dogs' BCS or dogs' being heavy/obese. An important and novel finding was that neutering increased the risk of being overweight or obese for male dogs while bitches were at risk irrespective of neuter status. Furthermore, a complex interaction between owners' weight status, feeding practices and the risk of dogs being overweight or obese was found, which stresses the need to consider companion animal obesity from a One Health perspective in future prospective studies. Finally, this study was unable to confirm that canine obesity is a product of owners being too attached to their dogs., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

The purpose of this study was to improve the diagnostic recommendations for Staphylococcus aureus and Streptococcus agalactiae control using bacterial culture (BC), polymerase chain reaction (PCR) and somatic cell count (SCC) as diagnostic methods. The study was carried out in three steps: firstly, diagnostic test patterns for naturally infected quarters with Staph. aureus (24 quarters) and Strep. agalactiae (16 quarters) were created by sampling the quarters each day for 21 days and analysing the daily quarter milk samples using BC, PCR and SCC. Secondly, 30 mastitis experts were asked to group and describe the diagnostic test patterns and to establish a diagnosis for each group. The experts' statements regarding the groups they established were subsequently examined using qualitative content analysis to assign "infection types" to the statements. Lastly, the test performance was estimated for BC, PCR and SCC using generalised logistic regression models with the interpreted statements as a reference for infection. The experts mainly identified the Staph. aureus quarter-patterns as persistent infections, while some had more dynamic patterns. Strep. agalactiae quarter-patterns mainly involved persistent infection, yet some appeared hard to diagnose and were assigned to almost all different infection types, while experts did not agree on the interpretation. Estimates of Se for detection of Staph. aureus infection were 95.9% [93.7; 97.3] for BC, 99.5% [98.3; 99.8] for PCR, and 96.1% [94.0; 97.5] for SCC. The corresponding Sp estimates were 74.5% [65.7; 81.7], 66% [57.2; 73.8] and 43.7% [36.2; 51.5] for BC, PCR and SCC, respectively. The Se estimates of BC and PCR for Strep. agalactiae infection were 100% [83.5; 100] and 99.9% [99.6; 100], respectively, whereas the Se of SCC detecting Strep. agalactiae infection was only 34.3% [26.4; 43.3]. This indicated that Strep. agalactiae-positive BC and PCR test results were more important than SCC results to the experts when diagnosing a quarter as infected. The Sp estimates of BC, PCR and SCC for Strep. agalactiae infection were 99% [72.8; 100], 97.7% [62.1; 99.9], and 65.7% [56.7; 73.7], respectively. We conclude that PCR and BC are highly sensitive in the detection of persistent and new infections as defined by the experts, although the Se was not always 100%. An accepted lower Sp suggests that experts place less emphasis on false-positive results. We recommend that efforts are made to develop consistent terminology to characterise intramammary infections over time so that the course of infection can be taken into account at diagnosis., (Copyright © 2019 Elsevier B.V. All rights reserved.)

We present a questionnaire-based measure of four animal ethics orientations. The orientations, which were developed in light of existing empirical studies of attitudes to animal use and ethical theory, are: animal rights, anthropocentrism, lay utilitarianism, and animal protection. The two latter orientations can be viewed as variants of animal welfarism. Three studies were conducted in Denmark in order to identify the hypothesised orientations, evaluate their concurrent validity, and report their prevalence and relevance in animal-related opinion formation and behaviour. Explorative factor analysis (Study 1) and confirmative factor analysis (Study 2) successfully identified the four orientations. Study 2 revealed good measurement invariance, as there was none or very modest differential item functioning across age, gender, living area, and contrasting population segments. Evaluation of concurrent validity in Study 2 found that the orientations are associated with different kinds of behaviour and opinion when the human use of animals is involved in the hypothesised directions. In Study 3, a representative population study, the animal protection orientation proved to be most prevalent in the Danish population, and as in study 2 the four orientations were associated with different behaviours and opinions. Remarkably, the animal protection orientation does not lead to increased animal welfare-friendly meat consumption, the main reason for this being non-concern about the current welfare status of farm animals. We argue that the developed measure covers a wide range of diversity in animal ethics orientations that is likely to exist in a modern society such as Denmark and can be used in future studies to track changes in the orientations and to understand and test hypotheses about the sources and justifications of people's animal-related opinions and behaviours., Competing Interests: The authors have declared that no competing interests exist.

Background: Food insecurity and its consequences have not received much attention in the Nordic, social-democratic welfare states. This study reports the prevalence of low and very low food security in Denmark, identifies its socio-demographic determinants and examines its associations with eating-related and health-related outcomes., Methods: A cross-sectional survey with a mixed-mode response format (telephone interviewing or Internet). A disproportional stratified random sampling design was employed (N = 1877). Main measure was the 6-item gradient food security construct adapted from the US., Results: Prevalence of low and very low food security was 6.0% (95% CI:5.4-8.5%) and 2.4% (95% CI:1.3-3.3%), respectively. Prevalence was highest in households with disposable income below OECD's poverty threshold, households receiving benefits or disability pensions, and single-parent households. After socio-demographic adjustment, adults from low/very low food secure households had a higher probability of eating an unhealthy diet (women: adj.RR 2.82 P < 0.001; men: adj.RR 2.15 P < 0.01). Adults from low/very low food secure households had lower life satisfaction (women: adj.RR 0.49, P < 0.05; men: adj.RR 0.09 P < 0.001) and higher risk of psychological distress (women: adj.RR 2.42 P < 0.05; men: adj.RR 8.95 P < 0.001). Obesity was more prevalent in low/very low food secure women (RR 2.44 P < 0.05), but not after socio-demographic adjustment., Conclusion: Food insecurity in Denmark is associated with adverse factors such as unhealthy diet, obesity, life satisfaction, and psychological distress. It is important to widen food insecurity research to non-liberal welfare states since low/very low food security is negatively associated with unhealthy eating and other health indicators, even in a social-democratic welfare state.

Welfare Quality ® proposes a system for aggregation according to which the total welfare score for a group of animals is a non-linear effect of the prevalence of welfare scores across the individuals within the group. Three assumptions serve to justify this: (1) experts do not follow a linear reasoning when they assess a welfare problem; (2) it serves to prevent compensation (severe welfare problems hidden by scoring well on other aspects of welfare); (3) experts agree on the weight of different welfare measures. We use two sources of data to examine these assumptions: animal welfare data from 44 Danish dairy farms with Danish Holstein Friesian cows, and data from a questionnaire study with a convenience sample of 307 experts in animal welfare, of which we received responses from over 50%. Our main results were: (1) the total group-level welfare score as assigned by experts is a non-linear function of the individual animal welfare states within the group; (2) the WQ system does not prevent what experts perceive as unacceptable compensation; (3) the level of agreement among experts appears to vary across measures. Our findings give rise to concerns about the proposed aggregation system offered by WQ., Competing Interests: The authors declare no conflict of interest.

Using a survey conducted in four Nordic countries (Denmark, Finland, Norway and Sweden) in 2012, we investigated whether eating out in cafés and restaurants in these countries is best characterized as a workday pattern activity or a leisure activity; and, whether frequent eating at cafés and restaurants is related to socio-demographic factors and factors relevant to the organization of daily life. We found that eating out is not a fundamental part of everyday eating. It is something which takes place occasionally. This may be taken to suggest that eating out in the Nordic countries is primarily a leisure activity. However, while this is an accurate portrayal of Denmark and Norway, eating out in Finland and Sweden is somewhat more common and linked to work-life. This difference probably reflects contrasting historical-institutional paths: in Finland and Sweden a food culture emphasizing cooked lunches and dedicated public policies supporting the provisioning of lunches outside the home may have promoted eating out. Multivariate analysis revealed that eating out declines with age. An urbanization effect exists, as residence in a capital city increases the propensity to eat out. There were socio-economic differences in all countries. We hypothesized that education and status would not significantly explain eating out activity after people's gastronomic interest had been controlled for. However, education was statistically significant in Sweden and Norway, and surprisingly status was negatively associated with eating out in Sweden. As expected, eating out was positively associated with income, and it was more frequent among the higher salariat and the self-employed (although not in Norway)., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A number of dog breeds suffer from welfare problems due to extreme phenotypes and high levels of inherited diseases but the popularity of such breeds is not declining. Using a survey of owners of two popular breeds with extreme physical features (French Bulldog and Chihuahua), one with a high load of inherited diseases not directly related to conformation (Cavalier King Charles Spaniel), and one representing the same size range but without extreme conformation and with the same level of disease as the overall dog population (Cairn Terrier), we investigated this seeming paradox. We examined planning and motivational factors behind acquisition of the dogs, and whether levels of experienced health and behavior problems were associated with the quality of the owner-dog relationship and the intention to re-procure a dog of the same breed. Owners of each of the four breeds (750/breed) were randomly drawn from a nationwide Danish dog registry and invited to participate. Of these, 911 responded, giving a final sample of 846. There were clear differences between owners of the four breeds with respect to degree of planning prior to purchase, with owners of Chihuahuas exhibiting less. Motivations behind choice of dog were also different. Health and other breed attributes were more important to owners of Cairn Terriers, whereas the dog's personality was reported to be more important for owners of French Bulldogs and Cavalier King Charles Spaniels but less important for Chihuahua owners. Higher levels of health and behavior problems were positively associated with a closer owner-dog relationship for owners of Cavalier King Charles Spaniels and Chihuahuas but, for owners of French Bulldogs, high levels of problems were negatively associated with an intention to procure the same breed again. In light of these findings, it appears less paradoxical that people continue to buy dogs with welfare problems.

This study examined the extent to which Danish veterinary practices encounter financially limited clients and how different factors relating to the animal, the client and the veterinarian affect decisions to provide treatment for these clients. 300 small animal practices were invited to participate in an online survey. 195 participated, giving a response rate of 65 per cent. The results show that Danish small animal veterinary practices encounter clients with limited finances regularly: 33.8 per cent of them 3-4 times, 24.6 per cent 5-10 times and 19.5 per cent 1-2 times a month. Only around 9 per cent reported having a written practice policy on handling financially limited clients. Factors affecting decisions to treat include the severity and type of the animal's condition, the medical care needed and the client's expressed emotions. The propensity to treat is significantly higher in female veterinarians and in situations involving unborn animals. The overall conclusion is that small animal veterinary practices often provide treatment to clients who are not able to pay-far beyond what is legally required. This can be considered a major economic and psychological challenge for the practising veterinarians., (British Veterinary Association.)

How have eating patterns changed in modern life? In public and academic debate concern has been expressed that the social function of eating may be challenged by de-structuration and the dissolution of traditions. We analyzed changes in the social context and conduct of eating in four Nordic countries over the period 1997-2012. We focused on three interlinked processes often claimed to be distinctive of modern eating: delocalization of eating from private households to commercial settings, individualization in the form of more eating alone, and informalization, implying more casual codes of conduct. We based the analysis on data from two surveys conducted in Denmark, Finland, Norway and Sweden in 1997 and 2012. The surveys reported in detail one day of eating in representative samples of adult populations in the four countries (N = 4823 and N = 8242). We compared data regarding where, with whom, and for how long people ate, and whether parallel activities took place while eating. While Nordic people's primary location for eating remained the home and the workplace, the practices of eating in haste, and while watching television increased and using tablets, computers and smartphones while eating was frequent in 2012. Propensity to eat alone increased slightly in Denmark and Norway, and decreased slightly in Sweden. While such practices vary with socio-economic background, regression analysis showed several changes were common across the Nordic populations. However, the new practice of using tablets, computers, and smartphones while eating was strongly associated with young age. Further, each of the practices appeared to be related to different types of meal. We conclude that while the changes in the social organization of eating were not dramatic, signs of individualization and informalization could be detected., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background/objectives: Daily practices related to eating are embedded in the social and cultural contexts of everyday life. How are such factors associated with diet quality relative to motivational factors? And, are associations universal or context-specific? We analyze the relationship between diet quality and the following practices: social company while eating, the regularity and duration of eating and the activity of watching TV while eating., Subjects/methods: A cross-sectional, questionnaire-based internet survey was conducted in April 2012 with stratified random samples of the populations (aged 15-80 years) in Denmark, Finland, Norway and Sweden (N=7531, completion rate 9-13%). The questionnaire elicited detailed accounts of one day of eating focusing on social and practical aspects of eating events. The validated Dietary Quality Score was the dependent variable. This measure is based on eight food frequency questions focusing on fats, vegetables, fruits and fish in the diet., Results: Eating activities were associated with diet quality even when motivation to eat healthily and sociodemographic factors were controlled for. The number of daily eating events and eating main meals was positively correlated with diet quality in all countries. Beyond that, activities that were significantly associated with diet quality varied with country. When measured separately, the association between each activity and diet quality was weaker than motivation to eat healthily, but in combinations that are found in parts of the populations, the association was substantial., Conclusions: Daily practices related to eating are correlated with diet quality. Practices that are important are in part universal but also country-specific. Efforts to promote healthy eating should address not only cognitive factors but also everyday contexts of eating that facilitate or hamper healthy practices.

Background/objectives: This study examined public support for publicly funded treatment of obesity (weight-loss surgery and medical treatment) and two pulmonary diseases (chronic obstructive pulmonary disease (COPD) and lung cancer) in Denmark. It also investigated whether beliefs about the causes of lifestyle-related diseases (external environment, genetic disposition and lack of willpower) and agreement that 'people lack responsibility for their life and welfare' influenced support., Subjects/methods: This was a questionnaire study in which a sample of 1003 Danes (age 18-65 years) drawn from an Internet database were surveyed., Results: Approximately one in three supported publicly funded weight-loss surgery (30%) and medical treatment of obesity (34.4%). A large majority supported treatment for lung cancer (86.1%), and a clear majority also supported treatment for COPD, whether it was framed as 'smoker's lung' (61.9%) or COPD (71.2%). The belief that lifestyle-related diseases are caused by the external environment or genetic disposition did not systematically influence support. Agreement that 'people lack responsibility for their life and welfare' reduced support significantly for all treatment types. However, in contrast with pulmonary diseases, support for publicly funded obesity treatments decreased considerably when beliefs about individual failure (that is that people lack 'willpower' and 'individual responsibility') were detected., Conclusions: Support for publicly funded COPD and lung cancer treatment is considerably higher than that for obesity treatment. This could encourage institutional discrimination through policies that involve charging patients for the treatment of obesity but not for the treatment of other lifestyle-related diseases.

There is a widely shared belief that contemporary eating culture is undergoing a process of 'destructuration' in which collective norms guiding temporal, social, and spatial aspects of eating as well as cuisine will decline or disappear. From another theoretical perspective one could argue that shared and regular patterns are quite resistant to change because they are functionally necessary for the organization and maintenance of social actions in everyday life. Using questionnaire data from Denmark, Finland, Norway and Sweden from the years 1997 and 2012 we investigate whether culturally shared timing of eating rhythms has disappeared or declined. At the population-wide level, we find clear national peaks (occurring around breakfast, lunch, and to a lesser extent dinner) during which a great number, or even the majority, of people eat. These basic rhythms of eating are nationally specific and clearly identifiable in 1997 and 2012, and only small changes were found to have occurred over the period studied. Subsequent examination of within-country differences in daily eating rhythms identified a specific sub-population with very similar features in all countries. The sub-population deviates temporally from the collective peaks of eating, and in it conventional meal types such as breakfast and lunch are skipped to a higher extent, giving what we call an 'unsynchronized' eating pattern. Interestingly, the pattern has become more common in all countries. While the growth of this sub-population may be a sign of a coming destructuration of meal culture, further analysis suggests this is not the case. Thus, we find clear socio-structural explanations for unsynchronized eating. It is related to the social coordination of work, and unsynchronized eating tends to be abandoned over the life course: with the establishment of a family, and old age, people tend to synchronize their eating habits with collective activities in society. Coupling this with the relatively modest growth of the unsynchronized pattern, and bearing in mind that it is a minority phenomenon, encompassing approximately one quarter of the population in 2012, we argue that an all-encompassing temporal destructuration will not develop. Additional analysis shows that the idea of a simultaneous rupture of eating culture on several dimensions (temporal, social, spatial, manners, cuisine) is doubtful. Thus, although, to a higher extent, individuals with an unsynchronized eating rhythm lack "manners" and eat more unhealthily, they do not display a higher degree of destructuration in the social and spatial dimensions of eating. Indeed, unsynchronized eating leads to fewer daily eating events, which contradicts the 'grazing' theory altogether., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Animal use in medical research is widely accepted on the basis that it may help to save human lives and improve their quality of life. Recently, however, objections have been made specifically to the use of animals in scientific investigation of human obesity. This paper discusses two arguments for the view that this form of animal use, unlike some other forms of animal-based medical research, cannot be defended. The first argument leans heavily on the notion that people themselves are responsible for developing obesity and so-called 'lifestyle' diseases; the second involves the claim that animal studies of obesity's causes and therapies distract attention from preventive efforts. Drawing on both empirical data and moral reasoning, we argue that the relevant attributions of responsibility and claims about distraction are not plausible, and that, therefore, there is no reason to single out the use of animals in obesity research as especially problematic.

The conflict between animal costs and human benefits has dominated public as well as academic debates about animal research. However, surveys of public perceptions of animal research rarely focus on this part of attitude formation. This paper traces the prevalence of different attitudes to animal research in the public when people are asked to take benefit and cost considerations into account concurrently. Results from the examination of two representative samples of the Danish public identify three reproducible attitude stances. Approximately 30-35% of people questioned approved of animal research quite strongly, and 15-20% opposed animal research. The remaining 50% were reserved in their views. Further studies will ideally use the measure developed here to make possible relatively fine-grained comparisons and understandings of differences between populations and changes in attitudes over time.

The aim of this study was to investigate the Danish public's support for publicly funded obesity treatment and prevention. It was also examined whether levels of support could be explained by dislike of obese people and/or the belief that those who are obese are personally responsible for their condition. A representative survey of members of the Danish public (N = 1,141) was conducted using a web-based questionnaire. The survey was designed to assess attitudes to public funding for obesity-related health care, and to investigate the impact, on those attitudes, of dislike of obese people, the perceived controllability of obesity, self-reported BMI, and additional attitudinal and socio-demographic characteristics. Public funding of some obesity treatments, such as weight-loss surgery, attracted only limited public support. A majority of the Danish public did support "softer" treatment interventions and preventive initiatives. Attitudes to the treatment of obesity were clearly best predicted by the belief that individuals are personally responsible for their own obesity. Dislike of obese persons had no direct effect on the preference for collective treatment initiatives and only a small effect on support for publicly funded obesity prevention. The high level of disapproval for publicly funded obesity treatment should be cause for concern for decision makers aiming to ensure equal access to health care. Since it is the belief that obese people are personally responsible which explains this disapproval, strategies for challenging public opinion on this issue are discussed.

We have analysed the consequences of liposome mediated gene transfer into human primary epidermal keratinocytes and compared non-Epstein-Barr Virus (EBV) and EBV based expression vectors that carry the genes encoding human Growth Hormone (hGH) or Enhanced Green Fluorescent Protein (EGFP). Different kinetics between the non-EBV and EBV based vectors were revealed upon subcultivation of hGH transfected keratinocytes. The keratinocytes transfected with non-EBV based vector showed a rapid reduction in hGH production. Although the EBV based vector resulted in more stable expression, this was also reduced over time. Chromatin inactivation by deacetylation was investigated by treatment with sodium butyrate and found not to be the reason for the decreasing expression. Keratinocytes divided into subpopulations enriched for either stem cells or transit amplifying cells, based on beta1-integrin expression and function, do not differ significantly with respect to susceptibility to productive transfection. However, when the keratinocytes were transfected with the EGFP gene and sorted live by FACS into EGFP negative and positive populations, only the negative cells were capable of forming significant numbers of colonies. This is consistent with the observation that the ability to incorporate BrdU was dramatically reduced in the EGFP expressing population within 24-48 h post transfection indicating an almost complete cell cycle arrest. p53 levels were unaffected by the procedures, and the keratinocyte cell line HaCat, mutated in both p53 alleles, also shows a marked reduction in clonogenic potency upon transfection. There was a slight increase of TUNEL positive apoptotic nuclei in the positive population at early time points. However, the apoptotic index was still very low. When we measured the frequency of involucrin expressing cells, we found an increase in the productively transfected population over time indicating an initiation of terminal differentiation. In contrast to the transfected cultures, keratinocytes that were transduced using a retroviral vector showed no decrease in colony forming efficiency. In conclusion we find that transgene expressing cells from transfected cultures of epidermal keratinocytes undergo cell cycle arrest and initiate terminal differentiation by mechanisms which are independent of p53 levels.

A detailed analysis of the subcellular distribution of acyl-CoA esters in rat liver revealed that significant amounts of long-chain acyl-CoA esters are present in highly purified nuclei. No contamination of microsomal or mitochondrial marker enzymes was detectable in the nuclear fraction. C16:1 and C18:3-CoA esters were the most abundant species, and thus, the composition of acyl-CoA esters in the nuclear fraction deviates notably from the overall composition of acyl-CoA esters in the cell. After intravenous administration of the non-beta-oxidizable [(14)C]tetradecylthioacetic acid (TTA), the TTA-CoA ester could be recovered from the nuclear fraction. Acyl-CoA esters bind with high affinity to the ubiquitously expressed acyl-CoA binding protein (ACBP), and several lines of evidence suggest that ACBP functions as a pool former and transporter of acyl-CoA esters in the cytoplasm. By using immunohistochemistry, immunofluorescence microscopy, and immunoelectron microscopy we demonstrate that ACBP localizes to the nucleus as well as the cytoplasm of rat liver cell and rat hepatoma cells, suggesting that ACBP may also be involved in regulation of acyl-CoA-dependent processes in the nucleus.

Most disease-causing missense mutations in short-chain acyl-CoA dehydrogenase (SCAD) and medium-chain acyl-CoA dehydrogenase are thought to compromise the mitochondrial folding and/or stability of the mutant proteins. To address this question, we studied the biogenesis of SCAD proteins in COS-7 cells transfected with cDNA corresponding to two SCAD missense mutations, R22W (identified in a patient with SCAD deficiency) or R22C (homologous to a disease-associated R28C mutation in medium-chain acyl-CoA dehydrogenase deficiency). After cultivation at 37 degreesC the steady-state amounts of SCAD antigen and activity in extracts from cells transfected with mutant SCAD cDNAs were negligible compared with those of cells transfected with SCAD wild type cDNA, documenting the deleterious effect of the two mutations. Analysis of metabolically labeled and immunoprecipitated SCAD wild type and mutant proteins showed that the two mutant proteins were synthesized as the 44-kDa precursor form, imported into mitochondria and processed to the mature 41.7-kDa form in a normal fashion. However, the intramitochondrial level of matured mutant SCAD proteins decreased rapidly to very low levels, indicating a rapid degradation of the mutant proteins at 37 degreesC. A rapid initial elimination phase was also observed following cultivation at 26 degreesC; however, significantly higher amounts of metabolically labeled and immunoprecipitated mature mutant SCAD proteins remained detectable. This corresponds well with the appreciable steady-state levels of SCAD mutant enzyme activity observed at 26 degreesC. In addition, confocal laser scanning microscopy of immunostained cells showed that the SCAD mutant proteins were localized intramitochondrially. Together, these results show that newly synthesized SCAD R22W and R22C mutant proteins are imported and processed in the mitochondrial matrix, but that a fraction of the proteins is rapidly eliminated by a temperature-dependent degradation mechanism. Thermal stability profiles of wild type and mutant enzymes revealed no difference between the two mutants and the wild type protein. Furthermore, the turnover of the SCAD mutant enzymes in intact cells was comparable to that of the wild type, indicating that the rapid degradation of the mutant SCAD proteins is not due to lability of the correctly folded tetrameric structure but rather to elimination of partly folded or misfolded proteins along the folding pathway.

To characterize disease-causing mutations in the low density lipoprotein receptor (LDL-R) gene, COS cells are transfected with the mutant gene in an EBV-based expression vector and characterized by flow cytometry. Using antibodies against the LDL-receptor the amount of receptor protein on the cell surface is quantitated. The receptor activity is measured by incubating the cells with fluorescence labeled LDL (Dil-labelled LDL) at 37 degrees C and 4 degrees C. The transfected cells stained with anti-LDL-R antibodies can also be analysed by immunofluorescence microscopy allowing the study of the intracellular location of variants of the receptor. To evaluate these methods, we are analyzing four previously well-characterized LDL-R mutations, belonging to each of the classes 2 to 5. Preliminary data show that mutant genes belonging to class 3 and 4A give rise to receptor protein on the cell surface, but impaired LDL uptake, while mutant receptors belonging to class 2A and 5 can only be detected intracellularly. Expression of the class 2A mutation results in an ER staining pattern, whereas the class 5 mutation gives rise to an intracellular staining compatible with localization in the endosomal/lysosomal compartments. We conclude that this system is useful for a rapid functional analysis of newly discovered mutations in the LDL-R gene.

Medium-chain acyl-CoA dehydrogenase (MCAD) deficiency is a potentially lethal inherited defect in the beta-oxidation of fatty acids. By comparing the behaviour of five missense MCAD mutant proteins expressed in COS cells and in Escherichia coli, we can define some of these as "pure folding mutants." Upon expression in E. coli, these mutant proteins produce activity levels in the range of the wild-type enzyme only if the chaperonins GroESL are co-overproduced. When overexpressed in COS cells, the pure folding mutants display enzyme activities comparable to the wild-type enzyme. The results suggest that the MCAD mutations can be modulated by chaperones, a phenomenon that may influence the manifestation of the MCAD disease.

In situ techniques for the detection of specific chromosomes using centromeric probes and the decoration of entire chromosomes using chromosome painting are well established. However, in the deciphering of complex chromosomal aberrations it is valuable to be able to detect the centromere and the entire DNA of a specific chromosome in different colours simultaneously on the same metaphase. In this report we describe a combination of the primed in situ labelling (PRINS) technique and chromosome painting for simultaneous visualization of centromere-specific oligonucleotides and chromosome painting libraries. A key feature is that the denaturation step in the PRINS reaction is sufficient to keep the chromosomes denatured for chromosome painting. This means that PRINS and consecutive chromosome painting can be performed as a single procedure (PRINS-painting).