Perceptions of Cross-Cultural Challenges and Successful Approaches in Facilitating the Improvement of Equine Welfare.

Projects that aim to improve the welfare of equids worldwide usually involve people from different countries and cultures working together. Given that professionals involved with multi-stakeholder projects often work cross-culturally, this study examined their experiences regarding the challenges involved in, and their reflections on, how to work in a culturally sensitive way. Semi-structured interviews were conducted with 14 participants working in a total of 29 countries and analysed using thematic analysis. Key response themes emerged from the responses to questions covering the areas of perceptions of animal welfare, challenges working cross-culturally and embracing cultural sensitivity. The overriding theme regarding perceptions of animal welfare was that of barriers to animal welfare, under which emerged the subthemes of limited financial and material resources, limited understanding of the tenets of animal welfare, and attachment to traditional medicines and practices. Exploring the key challenges resulted in two themes: challenges regarding the local context and etiquette, and those regarding working with different stakeholders. Considering cultural sensitivity, again, two themes emerged: the importance of trust and respect, and of working with local partners. Previous works have highlighted the importance of shared linguistic knowledge, interpersonal skills and cultural knowledge, and these elements also emerged in this research. As well as providing insights into the challenges of working cross-culturally, the findings of this study have enabled the development of suggestions for how this work could be taken forward in a practical way to be of use to professionals in this sector.

'Using human behaviour change can aid more impactful interactions with clients'.

The team at Human Behaviour Change for Life believes that by using the science of human behaviour change the veterinary community has an opportunity to better engage with pet breeders regarding extreme conformation.

Perceptions of Fear and Anxiety in Horses as Reported in Interviews with Equine Behaviourists.

One of the key welfare concerns for horses in the United Kingdom is lack of recognition of fear in horses. This study aimed to gain an understanding of how well horse care givers recognise fear and/or anxiety in horses by interviewing equine behaviourists (who interact with large numbers of horse care givers and talk to them about this topic routinely). The experiences of Animal Behaviour and Training Council (ABTC)-registered equine behaviourists working with horse caregivers were examined, including the ability of clients to recognise fear and/or anxiety in horses, how clients respond when discussing fear as the reason for their horse's behaviour, and what explanations the participants use to explain fear and anxiety. Semi-structured interviews were conducted with nine participants and analysed using thematic analysis before being written up to reflect the discussion points. When asked how well horse caregivers recognise fear and/or anxiety in horses, three key response themes emerged: caregivers are extremely poor at recognizing fear and anxiety in horses; some clients do recognise behavioural signs indicating fear and/or anxiety but only the overt signs (e.g., rearing, running away) rather than the more subtle signs (e.g., tension in face, subtle avoidance behaviours such as a hesitant gait); and fear and/or anxiety behaviour is often misinterpreted or mislabelled. These key themes recurred throughout several other interview questions. This study has provided initial insights into the lack of recognition of fear and anxiety of horses by their caregivers in the United Kingdom as well as tried and tested approaches to conversations to change this. Such synthesis of experience and techniques across the equine behaviour sector, together with the information gained regarding perception of equine caregivers, could be a valuable approach to improve the effectiveness of behaviour consultations and welfare initiatives.

Attitudes of the Equestrian Public towards Equine End-of-Life Decisions.

A key welfare concern for the equine population in the U.K. has been identified as delayed death, leading to prolonged suffering of horses. Reasons why some horse owners fail to have their horses euthanised include financial cost, emotional attachment, peer pressure, negative attitudes towards killing and poor recognition of behavioural indicators of equine pain and stress. The Five Freedoms framework of welfare was used to build a Likert-style survey to investigate the factors underlying attitudes of horse owners towards welfare measures in an end-of-life decision. Participants were asked to respond to hypothetical welfare scenarios and to give details of any horses they had had euthanised. The survey was conducted predominantly via equestrian Facebook groups and obtained 160 participant responses. Reliability of the scale was acceptable, with Cronbach's α=0.89. Principal Component Analysis was used to load the hypothetical scenarios onto seven factors containing 62.2% of the variance. The first four factors could be categorized according to "Ethology-informed Management", "Traditional Horse Management", "Emotional Issues" and "Physical Issues". Participants were more likely to consider euthanasia for physical issues, compared with issues relating to affective state and/or ethology, although it was not clear whether this was due to disregard for welfare issues relating to mental health or failure to recognise them as such. A large number of responses stated that the scenario had no bearing on whether a horse should be euthanised, again suggesting a lack of recognition of welfare issues and their implications. When asked to state their reasons for euthanising their horses, participants cited almost exclusively physical reasons, with the exception of those citing dangerous behaviour. Only a small number of responses also included consideration of affective and/or ethological factors, suggesting that welfare issues concerning affective state and/or behaviour are at risk of omission from end-of-life decisions.

Response of UK Horse, Pony and Donkey Owners to the Early Stages of the COVID-19 Pandemic.

In the UK, March 2020 was a time of great uncertainty as COVID-19 became increasingly widespread. The government responded by making suggestions about how people could reduce the risk of spread on 16 March, moved swiftly into closing schools on the 18 March before announcing a mandatory lockdown on the 23rd March. This was a challenging time for UK equestrians who had to balance maintaining their equine's routine and daily care alongside the increasing biosecurity measures. A cross-sectional survey was distributed to UK equestrians via social media over two days (28 and 29 March 2020) to better understand the decisions made by UK horse, pony and donkey owners during this time. Data from 452 respondents were generated across all four countries comprising the UK, although there were no significant differences in owner response to the pandemic between locations. The changes respondents made differed between the 16th and the 18th of March 2020, with an early emphasis on improving yard biosecurity and opting to stop riding, as well as reducing the time spent at the yard. After the 18 March, respondents placed more emphasis on risk reduction by changing the activities they did with their horse, including riding, with common examples including avoiding "high risk" activities such as riding on busy roads, jumping, riding young or nervous horses. Few respondents reported having an emergency plan in place should they become ill or otherwise unable to care for their equine. The findings highlight areas that would significantly benefit from in-depth investigation in future research. Equestrian behaviour and mindset around risk-taking and risk perception have already been researched in relation to equestrian activities and sport but have received little attention in the context of wider health challenges. Understanding the uptake of emergency planning and preparation in the UK equestrian community also warrants consideration. Using this information effectively to promote forward planning is likely to be of great benefit in equestrian responses to future health or climate-related crises.

Improving the Recognition of Equine Affective States.

A key welfare problem for horses is that people commonly fail to recognise, and consequently neglect to resolve, equine behavioural signs of distress, worsening the welfare of the horse and potentially putting the safety of the handler at risk as a result. Members of equestrian Facebook groups were asked to view six videos and assess the horse's behaviour in each; the authors selected the videos and considered each video to demonstrate behaviour associated with negative affective states. An additional six equine behaviourists also completed the survey as an "expert comparison group" from whom we could define "correct" answers; their responses were consistent with each other and the views of the authors. Although the majority of respondents successfully recognised behaviour indicative of distress in some instances, behaviour associated with negative affective states was commonly missed; videos featuring natural horsemanship and bridle-less riding were particularly interpreted incorrectly to be positive experiences for the horses. Binary logistic regression analysis (72.1% success rate) confirmed that the different video types (ridden dressage, natural horsemanship, in-hand dressage, bridle-less riding, Western reining and behavioural rehabilitation) were strong predictors for obtaining a correct answer (p < 0.01) but that experience of equine-ownership was not. Of the equestrian activities preferred by participants, only proponents of clicker training showed an increased likelihood of obtaining the correct answer (p = 0.05). Even when behavioural signs suggestive of negative affective states were recognised, a minority of respondents stated that they would be happy for their horse to be treated similarly. In conclusion, behavioural signs of equine distress are poorly recognised; they therefore warrant an increased prominence in education and the outreach activity of welfare organisations, in order to reduce equine suffering.

Repurposing Drugs to Target the Diabetes Epidemic.

Despite major investment by pharmaceutical companies in conventional drug discovery pipelines, development of new drugs has failed to keep up with the increasing incidence of many diseases, including type 2 diabetes (T2D). Drug repurposing, where existing drugs are applied to a new indication, is gaining momentum as a successful approach to overcome the bottlenecks commonly encountered with conventional approaches. Repurposing takes advantage of available information on the molecular pharmacology of clinical agents to drastically shorten drug development timelines. This review discusses recent advances in the discovery of new antidiabetic agents using repurposing strategies.

Chronic heart failure selectively induces regional heterogeneity of insulin-responsive glucose transporters.

Glucose uptake across the sarcolemma is regulated by a family of membrane proteins called glucose transporters (GLUTs), which includes GLUT4 (the major cardiac isoform) and GLUT12 (a novel, second insulin-sensitive isoform). Potential regional patterns in glucose transport across the cardiac chambers have not been examined; thus, we hypothesized that insulin-responsive GLUT4 and -12 protein and gene expression would be chamber specific in healthy subjects and during chronic heart failure (HF). Using a canine model of tachypacing-induced, progressive, chronic HF, total GLUT protein and messenger RNA in both ventricles and atria (free wall and appendage) were investigated by immunoblotting and real-time PCR. In controls, GLUT4, but not GLUT12, protein content was significantly higher in the atria compared with the ventricles, with the highest content in the right atrium (RA; P < 0.001). GLUT4 and GLUT12 mRNA levels were similar across the cardiac chambers. During chronic HF, GLUT4 and GLUT12 protein content was highest in the left ventricle (LV; by 2.5- and 4.2-fold, respectively, P < 0.01), with a concomitant increase in GLUT4 and GLUT12 mRNA (P < 0.001). GLUT4, but not GLUT12, protein content was decreased in RA during chronic HF (P = 0.001). In conclusion, GLUT4 protein was differentially expressed across the chambers in the healthy heart, and this regional pattern was reversed during HF. Our data suggest that LV was the primary site dependent on both GLUT4 and GLUT12 during chronic HF. In addition, the paradoxical decrease in GLUT4 content in RA may induce perturbations in atrial energy production during chronic HF.

Evolutionary ancestry and novel functions of the mammalian glucose transporter (GLUT) family.

BACKGROUND: In general, sugar porters function by proton-coupled symport or facilitative transport modes. Symporters, coupled to electrochemical energy, transport nutrients against a substrate gradient. Facilitative carriers transport sugars along a concentration gradient, thus transport is dependent upon extracellular nutrient levels. Across bacteria, fungi, unicellular non-vertebrates and plants, proton-coupled hexose symport is a crucial process supplying energy under conditions of nutrient flux. In mammals it has been assumed that evolution of whole body regulatory mechanisms would eliminate this need. To determine whether any isoforms bearing this function might be conserved in mammals, we investigated the relationship between the transporters of animals and the proton-coupled hexose symporters found in other species. RESULTS: We took a comparative genomic approach and have performed the first comprehensive and statistically supported phylogenetic analysis of all mammalian glucose transporter (GLUT) isoforms. Our data reveals the mammalian GLUT proteins segregate into five distinct classes. This evolutionary ancestry gives insight to structure, function and transport mechanisms within the groups. Combined with biological assays, we present novel evidence that, in response to changing nutrient availability and environmental pH, proton-coupled, active glucose symport function is maintained in mammalian cells. CONCLUSIONS: The analyses show the ancestry, evolutionary conservation and biological importance of the GLUT classes. These findings significantly extend our understanding of the evolution of mammalian glucose transport systems. They also reveal that mammals may have conserved an adaptive response to nutrient demand that would have important physiological implications to cell survival and growth.

Utilising (14)C-radiolabelled atom transfer radical polymerisation initiators.

(14)C-radio-initiated atom transfer radical polymerisations allow direct monitoring of the fate of initiating species.

Octogenarians on dialysis: lessons in longevity and quality of life.

A review of literature indicates that people who experience longevity have certain characteristics and personality traits in common. While family history and health habits play an important role, centenarian research reveals that family support, an optimistic attitude, a sense of purpose, and an ability to manage stress, all can contribute to longevity. Social workers conducted intensive life review interviews with 17 octogenarians receiving dialysis at a clinic in the Detroit suburb Eastpointe, Mich. to learn more about their backgrounds, health habits, and coping styles in order to determine if they shared tendencies found in centenarians, and the implications it had for younger dialysis patients. Our initial findings support our hypothesis: Octogenarians on dialysis often share characteristics with centenarians, something that may help them to stave off or ameliorate the effects of chronic illness until later in life.

Mitogen-stimulated and rapamycin-sensitive glucose transporter 12 targeting and functional glucose transport in renal epithelial cells.

We hypothesized that glucose transporter 12 (GLUT12) is involved in regulation of glucose flux in distal renal tubules in response to elevated glucose. We used the Madin-Darby canine kidney polarized epithelial cell model and neutralizing antibodies to analyze GLUT12 targeting and directional GLUT12-mediated glucose transport. At physiological glucose concentrations, GLUT12 was localized to a perinuclear position. High glucose and serum treatment resulted in GLUT12 localization to the apical membrane. This mitogen-stimulated targeting of GLUT12 was inhibited by rapamycin, the specific inhibitor of mammalian target of rapamycin (mTOR). The functional role of GLUT12 was also examined. We constructed a GLUT12 cDNA containing a c-Myc epitope tag in the fifth exofacial loop. Assays of glucose transport at the apical membrane were performed using Transwell filters. By comparing transport assays in the presence of neutralizing anti-c-Myc monoclonal antibody, we specifically measured GLUT12-mediated glucose transport at the apical surface. GLUT12-mediated glucose transport was mitogen dependent and rapamycin sensitive. Our results implicate mTOR signaling in a novel pathway of glucose transporter protein targeting and glucose transport. Activity of the mTOR pathway has been associated with diabetic kidney disease. Our results provide evidence for a link between GLUT12 protein trafficking, glucose transport and signaling molecules central to the control of metabolic disease processes.

The power of collaboration with patient safety programs: building safe passage for patients, nurses, and clinical staff.

Patient safety is a relatively new field, with many opinions and few effectively proven approaches. One factor is clear: optimal patient safety outcomes cannot be achieved in isolation. Although it is well recognized that multidisciplinary collaboration in the healthcare setting is necessary to effect patient safety, collaboration with resources external to healthcare-academia and industry in particular-will not only aid but also quicken the patient safety efforts. The authors outline a healthcare system's use of all available resources to build a patient safety program.

Renal expression and localization of the facilitative glucose transporters GLUT1 and GLUT12 in animal models of hypertension and diabetic nephropathy.

Renal tubular glucose reabsorption is mediated by facilitative glucose transporter (GLUT) proteins and energy-dependent sodium glucose luminal transporters. Glucose transport in the diabetic kidney is upregulated and has been implicated in the pathogenesis of progressive diabetic nephropathy. Hyperglycemia, hypertension, and activation of the renin-angiotensin system are believed important in the development of the disease. The present study examines the renal expression of the facilitative glucose transporters GLUT1 and GLUT12 in rat models of diabetic nephropathy. Sprague-Dawley and transgenic (mRen-2)27 rats received either streptozotocin-induced diabetes or vehicle. GLUT12 expression and localization were determined by immunohistochemistry, immunoblotting, in situ hybridization, and confocal immunofluorescence. GLUT1 immunolabeling was detected on the basolateral membrane throughout the nephron. GLUT12 was localized to the distal tubules and collecting ducts. A significant increase in GLUT12 immunolabeling was measured in Ren-2 controls and Ren-2 diabetic animals compared with Sprague-Dawley controls. GLUT12 expression was higher in Ren-2 diabetic compared with Sprague-Dawley diabetic rats. Long-term diabetes resulted in significant increases in GLUT1 levels in the renal proximal tubules and expression was higher in Ren-2 diabetic than Sprague-Dawley diabetic rats. GLUT12 protein was localized to the cytoplasm and to the apical membrane of human and rat distal tubules and collecting ducts. The apical localization of GLUT12 in the distal tubules and collecting ducts suggests that it could contribute to additional glucose reabsorption in the late nephron. Levels of both GLUT1 and GLUT12 are elevated in animal models of hypertension and diabetic nephropathy.

Molecular and cellular regulation of glucose transporter (GLUT) proteins in cancer.

Malignant cells are known to have accelerated metabolism, high glucose requirements, and increased glucose uptake. Transport of glucose across the plasma membrane of mammalian cells is the first rate-limiting step for glucose metabolism and is mediated by facilitative glucose transporter (GLUT) proteins. Increased glucose transport in malignant cells has been associated with increased and deregulated expression of glucose transporter proteins, with overexpression of GLUT1 and/or GLUT3 a characteristic feature. Oncogenic transformation of cultured mammalian cells causes a rapid increase of glucose transport and GLUT1 expression via interaction with GLUT1 promoter enhancer elements. In human studies, high levels of GLUT1 expression in tumors have been associated with poor survival. Studies indicate that glucose transport in breast cancer is not fully explained by GLUT1 or GLUT3 expression, suggesting involvement of another glucose transporter. Recently, a novel glucose transporter protein, GLUT12, has been found in breast and prostate cancers. In human breast and prostate tumors and cultured cells, GLUT12 is located intracellularly and at the cell surface. Trafficking of GLUT12 to the plasma membrane could therefore contribute to glucose uptake. Several factors have been implicated in the regulation of glucose transporter expression in breast cancer. Hypoxia can increase GLUT1 levels and glucose uptake. Estradiol and epidermal growth factor, both of which can play a role in breast cancer cell growth, increase glucose consumption. Estradiol and epidermal growth factor also increase GLUT12 protein levels in cultured breast cancer cells. Targeting GLUT12 could provide novel methods for detection and treatment of breast and prostate cancer.

Agrobacterium-mediated transformation of the wetland monocot Typha latifolia L. (Broadleaf cattail).

An Agrobacterium-mediated model transformation system was standardized for the wetland monocot Typha latifolia L. to achieve the long-term objective of introducing candidate genes for phytoremediation. Two binary plasmid vectors, pCAMBIA1301/EHA105 and pTOK233/LBA4404, both containing the gus (beta-glucuronidase) and hptII (hygromycin phosphotransferase II) genes, were used for transformation. Fifty-day-old 5 mg/l picloram-derived calli were cocultivated and selected on medium containing 20 mg/l or 40 mg/l hygromycin. Resistant calli were regenerated on medium supplemented with 5 mg/l 6-benzylaminopurine, with or without 20 mg/l or 40 mg/l hygromycin and with or without charcoal (10 g/l). Transient GUS activity in explants ranged between 28% and 36%. Hygromycin-resistant calli, selected after 3 months, showed stable GUS expression. A total of 46 plants were regenerated and established in the greenhouse; 13 showed stable GUS expression. Cocultivation of dark culture-derived calli, directly selected on regeneration medium containing 20 mg/l hygromycin and rooted on medium with 20 mg/l hygromycin was the best protocol. The addition of charcoal did not have any effect on regeneration. PCR and Southern analyses of transgenic calli and transgenic plants confirmed the presence of the introduced genes. In conclusion, T. latifolia could be genetically transformed by Agrobacterium tumefaciens.

Glucose transporter GLUT12-functional characterization in Xenopus laevis oocytes.

We have recently identified and cloned the cDNA of a new member of the glucose transporter family that has been designated GLUT12. GLUT12 possesses the structural features critical to facilitative transport of glucose but the key to understanding the possible physiological roles of this novel protein requires analysis of functional glucose transport. In the current study, we have utilized the Xenopus laevis oocyte expression system to assay transport of the glucose analog 2-deoxy-D-glucose and characterize the glucose transport properties and hexose affinities of GLUT12. Our results demonstrate that GLUT12 facilitates transport of glucose with an apparent preferential substrate affinity for glucose over other hexoses assayed. The results are significant to understanding the potential role and importance of GLUT12 in insulin-sensitive tissues and also cells with high glucose utilization such as cancer cells.

Expression and localization of GLUT1 and GLUT12 in prostate carcinoma.

BACKGROUND: Increased glucose consumption is a characteristic of malignant cells and in prostate carcinoma is associated with the proliferation of both androgen-dependent and independent cells. Transport of polar glucose across the nonpolar membrane relies on glucose transporter proteins, known as GLUTs. Increased expression of GLUT1 is a characteristic of many malignant cells. The authors characterized and cloned the cDNA for a novel glucose transporter, GLUT12, which was identified initially in malignant breast epithelial cells. To the authors' knowledge, there have been no reports on the expression of glucose transporters in the human prostate or human prostate carcinoma cells. The authors evaluated GLUT1 and GLUT12 expression in human prostate carcinoma cells. METHODS: Reverse transcription-polymerase chain reaction was performed on total RNA extracted from cultured prostate carcinoma cells LNCaP, C4, C4-2, and C4-2B using primers to amplify GLUT1, GLUT12, or the housekeeping gene, 36B4. Total protein extracted from prostate carcinoma cell lines was assessed for GLUT12 protein by Western blot analysis. Cultured cell monolayers were incubated with antibodies to GLUT1 or GLUT12 and a peripheral Golgi protein, Golgi 58K, for detection by immunofluorescent confocal microscopy. Sections of benign prostatic hyperplasia and human prostate carcinoma were stained for immunohistochemical detection of GLUT1 and GLUT12. RESULTS: GLUT1 and GLUT12 mRNA and protein were detected in all cell lines evaluated. Immunofluorescence staining demonstrated both GLUT1 and GLUT12 on the plasma membrane and in the cytoplasm in all cultured prostate carcinoma cell lines, with GLUT1 but not GLUT12 appearing to colocalize with the Golgi. Immunohistochemical staining of benign prostatic hyperplasia indicated expression of GLUT1 but not GLUT12. Malignant tissue stained for GLUT12 but was negative for GLUT1. CONCLUSIONS: GLUT1 and GLUT12 are expressed in human prostate carcinoma cells. One possible rationale for the GLUT1 Golgi association is that it may supply glucose to the Golgi for byproduct incorporation into the prostatic secretory fluid. Further work will investigate the importance of glucose transport and GLUT1 and GLUT12 in prostate carcinoma cell growth.