Inflammatory markers for differentiation of endometritis in the mare.

BACKGROUND: Endometrial biopsy is required to diagnose mares with chronic endometritis and endometrial degenerative fibrosis. An increase in understanding of equine reproductive immunology could be utilised to create less-invasive, time-efficient diagnostic tools especially when evaluating mares for chronic endometritis. OBJECTIVES: To evaluate inflammatory cytokine and chemokine concentrations in uterine fluid samples collected by low-volume lavage (LVL) as a potential screening diagnostic biomarker for endometritis. STUDY DESIGN: Prospective cross-sectional clinical study. METHODS: Forty-six mares underwent a LVL and subsequently endometrial biopsy. Mares were split in three groups: healthy, acute endometritis, and chronic endometrial fibrosis (CEF) based on cytological and histological evaluation. A fluorescent bead-based multiplex assay for IFN-γ, IFN-α, IL-1ß, IL-4, IL-10, IL-17, sCD14, TNF-α, CCL2, CCL3, CCL5 and CCL11 were carried out on the LVL fluid. The endometrial biopsy was utilised for histology and qPCR of IFN-γ, IL-1ß, IL-6, IL-8, IL-17, TNF-α, CCL2 and CCL3 genes. Statistical analyses examined differences in inflammatory markers and predictive modelling for diseased endometrium. RESULTS: Secreted concentrations of IFN-γ were lower in LVL fluid from reproductively healthy mares compared with acute endometritis (p = 0.04) and CEF (p = 0.006). Additionally, IL-17, IL-10, IL-1ß, TNF-α, CCL2, CCL3, CCL5 and CCL11 were significantly increased (p ≤ 0.04) in LVL from CEF mares compared with healthy mares. Mares with CCL2 concentrations ≥550 pg/mL (14/14) had 100% probability of having CEF and/or acute endometritis. Healthy mares had lower relative abundance of IL-17 mRNA compared with mares in CEF group [median (interquartile rage) = 14.76 (13.3, 15.3) and 12.4 (10.54, 13.81)], respectively (p = 0.02). MAIN LIMITATIONS: Limited sample size: larger numbers of mares with and without endometritis are required and reference intervals in LVL samples have to be established. CONCLUSIONS: Inflammatory chemokines and cytokines concentrations differed between healthy mares and mares with acute endometritis or CEF in LVL.

Rho-associated coiled-coil containing kinase inhibitor improves outcomes of direct-transfer slow-cooled bovine blastocysts.

Direct-transfer slow-cooling cryopreservation is a widely used method for bovine embryo cryopreservation. However, the transfer of cryopreserved embryos is associated with reduced pregnancy rates. Rho-associated coiled-coil containing kinase inhibitor (ROCKi) has shown promise in improving the viability of post-warmed vitrified bovine embryos. Our objective was to investigate the effects of ROCKi treatment prior to slow-cooling or after cryopreservation on embryo viability. In vitro produced bovine embryos (n = 571) were randomly assigned to one of five groups: No-cryopreservation control group (NC-C), C-C group were cryopreserved by slow-rate cooling without ROCKi at any point, R-C group were incubated with ROCKi for 2 h before cryopreservation, C-R group were not exposed to ROCKi prior to cryopreservation but were cultured with ROCKi after cryopreservation, and R-R group were exposed to ROCKi before and after cryopreservation. Treatment group was significantly associated with blastocoel re-expansion, hatching, and degeneration (P < 0.0001). Blastocoel re-expansion rates were lower (P < 0.05) in the C-C (75.2 ± 4.2%) and R-C (85.2 ± 4.7%) groups compared with the NC-C (99.0 ± 0.7%), C-R (94.7 ± 2.6%) and the R-R (94.5 ± 2.9%) groups. The median time to re-expansion was significantly slowest in the C-C group (650, 560-915 min), followed by the R-C group (538, 421-611 min), then the C-R and R-R groups were similar (291, 261-361 and 321, 271-371 min) and the NC-C group was the fastest (196, 161-230 min) (P < 0.05). Similarly, the post-thaw hatching rate was lower, and the median time to hatching slower in the C-C (58.1 ± 7.0%, 2,033, 1634-2820 min) and R-C (65.7 ± 6.9%, 1,853, 1494-2356 min) groups compared with the NC-C (81.7 ± 6.0%, 1,309, 1084-1514 min), C-R (77.2 ± 6.5%, 1,384, 1013-1754 min) and R-R (82.0 ± 5.3%, 1,209, 943-1424 min) groups. ROCKi supplementation after cryopreservation resulted in fewer degenerated embryos (C-R = 8.9 ± 2.8%, and R-R 7.1 ± 2.8%) compared to the C-C (26.8 ± 4.3%) and R-C (17.9 ± 5.7%) groups. Exposure to ROCKi both before cryopreservation and after-cryopreservation yielded the best outcomes, similar to NC-C control group without cryopreservation, and significantly better than the C-C control group without supplements. Exposure to ROCKi after cryopreservation demonstrated greater benefits compared to exposure before cryopreservation alone. These findings suggest that ROCKi can potentially enhance cryosurvival of bovine embryos.

"Excess" Doubling Up During COVID: Changes in Children's Shared Living Arrangements.

The proportion of U.S. children living in doubled-up households, in which a child lives with a parent plus adult kin or nonkin, has increased in the last 40 years. Although shared living arrangements are often understood as a strategy to cope with crises, no research to date has examined changes in children's living arrangements during the first year of the COVID-19 pandemic. We use the American Community Survey and the Current Population Survey to examine children's doubled-up living arrangements during 2020 and the extent to which children may have experienced "excess" doubling up relative to earlier years. We consider trends by household type (multigenerational, extended with other relatives, and nonrelative households) and differences by demographic characteristics (marital status, race and ethnicity, work status, education, age, and number of coresident children). We find evidence that more than half a million (509,600) children experienced "excess" doubling up in 2020. Greater than expected increases in doubled-up arrangements were driven by increases in multigenerational households, in particular among Black and Hispanic children, young children (under age six), those whose mothers never married, and those whose mothers were not working. Correlates of coresidence remained largely unchanged over time, although having a mother who had never married became a stronger correlate in 2020. Our findings suggest that both economic and instrumental needs likely explained the rise in multigenerational coresidence in 2020.

Anti-Leishmania amazonensis Activity, Cytotoxic Features, and Chemical Profile of Allium sativum (Garlic) Essential Oil.

Human tegumentary leishmaniasis (HTL) is a serious tropical disease caused by Leishmania amazonensis. Developing new leishmanicidal agents can help overcome current treatment challenges, such as drug resistance and toxicity. Essential oils are a source of lipophilic substances with diverse therapeutic properties. This study aimed to determine the anti-L. amazonensis activity, cytotoxicity, and chemical profile of Allium sativum essential oil (ASEO). The effect of ASEO on parasite and mammalian cells viability was evaluated using resazurin and MTT assays, respectively. The oil's effect against intracellular amastigotes was also determined. Transmission electron microscopy was used to assess the ultrastructural changes induced by ASEO. In addition, the chemical constituents of ASEO were identified by gas chromatography-mass spectrometry (GC-MS). The cytotoxic potential was evaluated in vitro and in silico. The oil displayed IC50 of 1.76, 3.46, and 3.77 µg/mL against promastigotes, axenic, and intracellular amastigotes, respectively. Photomicrographs of treated parasites showed plasma membrane disruption, increased lipid bodies, and autophagic-like structures. ASEO chemical profiling revealed 1,2,4,6-tetrathiepane (24.84%) and diallyl disulfide (16.75%) as major components. Computational pharmacokinetics and toxicological analysis of ASEO's major components demonstrated good oral bioavailability and better toxicological endpoints than the reference drugs. Altogether, the results suggest that ASEO could be an alternative drug candidate against HTL.

Quality of Life of Mothers and Fathers 4 to 6 Months After Birth: The Effect of a Very Preterm Delivery.

OBJECTIVES: The sparse literature on the effect of a preterm delivery on parents' quality of life (QoL) yields inconsistent results, restricting their analysis to mothers. The present study aimed to assess the effect of a very preterm delivery on parents' gender-specific perception of QoL, 4 to 6 months after birth. METHODS: A total of 117 parents of very preterm infants hospitalized at birth in a level III Neonatal Intensive Care Unit, and 214 parents of never hospitalized full-term infants born in a public maternity, both located in the North of Portugal, participated in the study, 4 to 6 months after delivery (November 2013-June 2015). The Portuguese version of the World Health Organization Quality of Life - BREF Inventory was applied and scores were transformed to reflect a 0 to 100 scale. RESULTS: The mean [standard deviation (SD)] of overall QoL ranged between 72.1 (13.3) among mothers of full-term infants and 74.6 (12.5) among mothers of very preterm infants. The perception of QoL was not significantly different among parents of very preterm and full-term infants, according to gender. The highest scores were observed in the psychological and physical dimensions, for both mothers and fathers. CONCLUSIONS FOR PRACTICE: The lack of differences on the perception of QoL among mothers and fathers of very preterm and full-term infants, highlights the need to deeply understand and explore the influence of accommodation mechanisms, the extended family/community and health policies on parental QoL trajectories.

The heterogeneous associations of universal cash-payouts with breastfeeding initiation and continuation.

Existing health literature documents the benefits of breastfeeding for the first six months of life. Prior research on barriers to breastfeeding has focused on the role of hospital initiatives, return to work, and individual mothers' characteristics. This study uses data from Alaska's Pregnancy Risk Assessment Monitoring System and the Alaska Permanent Fund Dividend, to investigate whether universal income support shapes mothers' breastfeeding behaviors. We find that payouts are associated with increases in breastfeeding initiation and short-term continuation (three months) among a sample of urban Alaskan mothers. These associations differ across mothers' socioeconomic and demographic characteristics (i.e., education, economic status, race, marital status). We contend that this type of income intervention may complement existing efforts to promote breastfeeding by removing financial barriers to breastfeeding.

Patterns of childhood adversity and health outcomes in early adolescence: Results from the Generation XXI cohort.

This longitudinal population-based birth cohort study aims to identify childhood adversity patterns over the first 13 years of life and to examine its association with health-related behaviours and outcomes in early adolescence. Using data from the Portuguese birth cohort Generation XXI, we performed latent class analysis to explore the underlying patterns of adversity from birth to early adolescence, using 13 adversity items assessed in five time points. Health-related behaviours and outcomes were evaluated at 13 years. Adjusting for parental unemployment, logistic regression models were performed to determine the association between adversity patterns and outcomes. Among 8647 participants, three adversity patterns were identified: "low adversity" (56.1%), "household dysfunction" (17.2%) and "multiple adversities" (26.7%). For the "household dysfunction" pattern, girls and boys showed associations with increased odds of alcohol/tobacco use (adjusted odds ratio [AOR]: 1.78; 95% confidence interval [CI]: 1.32-2.40; AOR:1.84; CI:1.38-2.46, respectively) and depressive symptoms (AOR:2.34; CI:1.58-3.48; AOR:5.45; CI:2.86-10.38, respectively). Boys also presented low consumption of fruits/vegetables (AOR:1.51; CI:1.04-2.19). For the "multiple adversities" pattern, both girls and boys showed an increased probability of alcohol/tobacco use (AOR:1.82; CI:1.42-2.33; AOR:1.63; CI:1.30-2.05, respectively) and depressive symptoms (AOR:3.41; CI:2.46-4.72; AOR:5.21; CI:2.91-9.32, respectively). Boys also revealed increased odds of low consumption of fruits/vegetables (AOR:1.67; CI:1.24-2.23). Childhood adversity patterns are associated with unhealthy behaviours and depressive symptoms in early adolescence. Public policies and early interventions targeting vulnerable children, families and communities can potentially reduce the detrimental effects of adversities on health and promote individual and community resilience.

Characterization of serum and tissue oxytocinase and tissue oxytocin in the pregnant and non-pregnant mare.

Oxytocin is a hormone with functions in: reproduction, maternal bonding, milk ejection, and feeding/social behavior, and is reported to be present in a variety of tissues. Our goal is to characterize oxytocin and leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase), a key regulator of oxytocin in mares. We measured serum and tissue LNPEP by ELISA from ovulation (D0) until D21-22 in non-pregnant (n = 5) and pregnant mares (n = 6); and in periparturient and postpartum mares (n = 18). Placenta (n = 7) and homogenized tissue of diestrus mares (n = 6) were evaluated using protein determinations and LNPEP ELISAs. Identification of LNPEP and OXT protein in tissues was also performed via western blot, immunohistochemistry and liquid chromatography-mass spectrometry (LC-MS/MS). Furthermore, in situ hybridization was performed for LNPEP and OXT on endometrium, myometrium, pituitary and corpus luteum (CL). Serum LNPEP concentration were similar. Placental LNPEP U/mg protein was highest in the body and pregnant horn. The highest to lowest LNPEP U/mg protein by tissue were: myometrium > follicle wall > endometrium > kidney > CL > liver. Oxytocin was identified in the equine pituitary, CL and placenta and is likely to act in autocrine or paracrine manner, while LNPEP may act systemically and locally to regulate the availability of OXT.

Serum progesterone and oxytocinase, and endometrial and luteal gene expression in pregnant, nonpregnant, oxytocin, carbetocin and meclofenamic acid treated mares.

Our objectives were to examine changes in endometrial and luteal gene expression during estrus, diestrus, pregnancy and treatments to induce luteolysis and putatively induce luteostasis. Groups were: Diestrus (DIEST), Estrus (ESTR), Pregnant (PREG), Oxytocin (OXY), Carbetocin (CARB), and Meclofenamic acid (MFA). Blood was obtained from day (D)12 to D15 for measurement of oxytocinase, also referred to as leucyl-cysteinyl aminopeptidase (LNPEP) and progesterone. Luteal biopsies were obtained on D12 and D15 and an endometrial biopsy on D15. Real-time RT-PCR was performed for the following genes: PGR, ESR1, OXTR,OXT, LNPEP, PTGS2, PTGFR, PLA2G2C, PTGES, SLC2A4, and SLC2A1. Regarding serum LNPEP, PREG and OXY (p-value<0.001) had higher concentrations than DIEST mares. Endometrial PTGES expression was higher (p-value <0.04) in DIEST, PREG and OXY than other groups. Endometrium from ESTR had increased expression of OXT (p-value < 0.02) compared to MFA and OXY mares. Carbetocin treatment: decreased serum progesterone and LNPEP; increased endometrial PLA2G2C; decreased endometrial PTGES; and decreased luteal aromatase and PTGES. Treatment with MFA: decreased endometrial PLA2G2C, increased endometrial PTGES; and resulted in less OXTR and OXT luteal abundance on D12 compared to D15. Endometrial and luteal expression of LNPEP is affected by physiologic stage and treatment and is involved in luteal function and pregnancy recognition pathways through effects on oxytocin and prostaglandin synthesis in the horse.

Antileishmanial Screening, Cytotoxicity, and Chemical Composition of Essential Oils: A Special Focus on Piper callosum Essential Oil.

Leishmania amazonensis is the etiological agent of tegumentary leishmaniasis, a disease characterized by the emergence of cutaneous and mucocutaneous ulcerated lesions that can evolve into severe destruction of skin tissue. Treatment of the disease is often accompanied by high toxicity and variable efficacy. Essential oils stand out for having diverse pharmacological properties. Here, we screened a panel of fourteen essential oils for their anti-L. amazonensis activity, cytotoxicity, and chemical profile. Lippia sidoides (LSEO) and Piper callosum (PCEO) oils displayed the best anti-promastigote and anti-amastigote activities with IC50 of 31 and 21 µg/ml, respectively. PCEO was the safest oil with a desirable selectivity index >10. In addition, PCEO showed no cytotoxicity against the VERO line and erythrocytes. PCEO-treated amastigotes displayed mitochondrial membrane depolarization and high levels of intracellular ROS. Safrole (54.72 %) was the main component of PCEO. The results described here highlight the use of essential oils to combat tegumentary leishmaniasis.

Endometrial and luteal gene expression of putative gene regulators of the equine maternal recognition of pregnancy.

Our understanding of the temporal changes in endometrial and luteal gene transcripts related to the actions of oxytocin and prostaglandin during early equine pregnancy is incomplete. Additionally, the role of oxytocinase, also known as Leucyl-cystinyl aminopeptidase (LNPEP), during early pregnancy in mares has not been previously investigated. Luteal and endometrial biopsies were obtained on Day (D)8, D10, D12 and D15 post-ovulation in pregnant (PREG) and diestrus (DIEST) mares for real-time qPCR. Differences in endometrial gene expression occurred over time in: SLC2A4, SLC2A1, PTGES, OXTR and LNPEP. PTGFR and PLA2G2C had lower relative abundance in PREG D15 endometrium compared to D10. OXT and OXTR were increased on D10 and 15 PREG, respectively. Regarding luteal mRNA relative abundance, ESR1, PTGS2, PTGFR, and PTGES had higher relative abundance in D12 of DIEST and PREG. Luteal expression of OXTR and OXT had higher relative abundance in D15 compared to D8, and LNPEP had higher relative abundance in D10 and 12. Endometrial and luteal PTGES had an increased mRNA abundance in both D12 DIEST and PREG mares, which may lead to additional luteoprotective prostaglandin E2 (PGE2) secretion. Furthermore, luteal SLC2A1 had higher relative abundance in pregnancy, and likely supports the high metabolic activity of luteal tissue by increasing glucose uptake. Oxytocinase is present in endometrial and luteal tissue and its role in oxytocin induced prostaglandin secretion is uncertain.

Prevalence of Adverse Childhood Experiences in the First Decade of Life: A Study in the Portuguese Cohort, Generation XXI.

Adverse childhood experiences (ACEs) are a modifiable risk factor for diseases throughout life. This study estimates the prevalence of ACEs in children, addressing associated sociodemographic characteristics and examining the relationship of ACEs with the child's health and behaviors. We used information on 5295 participants at 10 years old, of the birth cohort Generation XXI, established in Porto, Portugal. Children answered a self-administered questionnaire on ACEs, based on the original ACEs study. Principal component analysis was used to group correlated ACEs, and a score was computed to assess their cumulative effect. Overall, 96.2% of children reported having been exposed to at least one ACE. The most prevalent ACE was a household member shouting, yelling, or screaming at the child (57.7%). Boys were more likely than girls to report "abuse", "school problems", and "death/severe disease". Low parental education, income, and unemployment were associated with an increased risk of "school problems", "death/severe disease", and "household dysfunction". We observed that the dimensions of ACEs could be identified at 10 years of age. A disadvantaged socioeconomic environment was associated with dimensions of ACEs. These data illustrate the natural history of dimensions of ACEs and their potential social patterning.

Benefits and Risks of Sharing Genomic Data for Research: Comparing the Views of Rare Disease Patients, Informal Carers and Healthcare Professionals.

Assessing public and patients' expectations and concerns about genomic data sharing is essential to promote adequate data governance and engagement in rare diseases genomics research. This cross-sectional study compared the views of 159 rare disease patients, 478 informal carers and 63 healthcare professionals in Northern Portugal about the benefits and risks of sharing genomic data for research, and its associated factors. The three participant groups expressed significantly different views. The majority of patients (84.3%) and informal carers (87.4%) selected the discovery of a cure for untreatable diseases as the most important benefit. In contrast, most healthcare professionals revealed a preference for the development of new drugs and treatments (71.4%), which was the second most selected benefit by carers (48.3%), especially by the more educated (OR (95% CI): 1.58 (1.07-2.34)). Lack of security and control over information access and the extraction of information exceeding research objectives were the two most often selected risks by patients (72.6% and 50.3%, respectively) and carers (60.0% and 60.6%, respectively). Conversely, professionals were concerned with genomic data being used to discriminate citizens (68.3%), followed by the extraction of information exceeding research objectives (54.0%). The latter risk was more frequently expressed by more educated carers (OR (95% CI): 1.60 (1.06-2.41)) and less by those with blue-collar (OR (95% CI): 0.44 (0.25-0.77) and other occupations (OR (95% CI): 0.44 (0.26-0.74)). Developing communication strategies and consent approaches tailored to participants' expectations and needs can benefit the inclusiveness of genomics research that is key for patient-centred care.

Expression of Oxytocin/Neurophysin I and Oxytocinase in the Equine Conceptus from Day 8 to Day 21 Post-Ovulation.

Leucyl and cystinyl aminopeptidase (LNPEP/oxytocinase) is an enzyme that metabolizes oxytocin in serum and tissues. The presence of oxytocin/neurophysin I (OXT), oxytocin and LNPEP and their relationship to other genes is unknown in the equine conceptus. Our objective was to characterize gene expression of LNPEP and OXT on D8, 10, 12, 14, 15, 16 and 21 conceptuses in relationship to other genes. Immunohistochemistry, western blot and liquid chromatography with tandem mass spectrometry (LC-MS/MS) were used for identification of oxytocin and LNPEP in D15, 16 and 18 conceptuses. LNPEP was increased at D15 compared to D10, was immunolocalized in the equine trophectoderm and endoderm, and protein was confirmed by LC-MS/MS. Maximal abundance of OXT was at D21, and lowest on D12 and D14, but no protein was identified. OXTR abundance was highest on D14 and D21. LNPEP was correlated with PTGFR and PTGES on D12 and D14-D15, and high expression of PTGES, PTGS2 was found on D14, D15 and D21; PTGFR was found on D8 and D12-21. LNPEP may have a role in prostaglandin regulation and conceptus fixation by decreasing the availability of oxytocin. Further investigation on the role embryonic LNPEP during pregnancy is warranted.

A scoping review on studies about the quality of life of informal caregivers of stroke survivors.

PURPOSE: To assess the available evidence regarding the quality of life (QoL) of informal caregivers of stroke survivors, by identifying the instruments used to assess QoL, and its associated characteristics. METHODS: A scoping review was performed, following PRISMA-ScR guidelines. The electronic databases PubMed, ISI Web of Science, PsycINFO, and SciELO were searched for empirical, peer-reviewed, original, and full-length studies on the characteristics influencing the QoL of informal caregivers of stroke survivors. Eligibility and data extraction were conducted by two independent researchers. The main quantitative findings were synthesized, and qualitative data were explored by thematic content analysis. RESULTS: The included studies, 56 quantitative, 1 qualitative, and 1 mixed methods, were published between 1999 and 2020. A high heterogeneity was found regarding the assessment of QoL, and the characteristics influencing it. Only one study used an instrument specifically designed to assess the stroke caregivers' QoL. The QoL of informal caregivers was inversely associated with physical and mental health of stroke survivors and caregivers, while stroke characteristics with a better prognosis, caregivers' positive relationships, and a more supportive and participative social context were positively associated to QoL. CONCLUSION: There is a need for standardizing the assessment of the QoL of informal caregivers of stroke survivors, as well as for investing in cross-country/cultural studies with robust mixed methods designs to allow a deeper understanding of the experiences of caregivers. Further research, policies, and practices should consider the diversity and complexity of the characteristics influencing QoL, to empower informal caregivers and improve their QoL.

Incidence and risk factors for canine mastitis and metritis in two guide dog populations.

Canine mastitis and metritis can cause severe illness but the incidence and risk factors have not been well-studied. Goals in the present study were: 1) report the incidence of mastitis and metritis in a large population, and 2) identify potential risk factors that predispose females to those diseases. A retrospective cohort study was conducted using data from two guide dog colonies that was collected for 17 and 10 years, respectively, for the two colonies. A total of 3076 whelpings occurred during the respective study periods and data were analyzed. Clinical mastitis was diagnosed in 13.2 % of whelpings (408 cases) with the average day of diagnosis being 16.7 postpartum. Risk factors for mastitis identified were colony, litter size where bitches that had large litter size of ≥9 pups (16.2 %) were 60 % more likely to develop mastitis compared with bitches that had litters of <9 pups (11.3 %). Bitches with congestion of the mammary gland were 4.8 times more likely to develop mastitis compared with bitches without mammary congestion. Case incidence of metritis was small (0.7 % of whelpings) and occurred on average at day-5 postpartum (range 1-16). There were no significant risk factors identified, and this may be due to the small number of metritis cases (22 cases) in the present study. Interpretations regarding metritis, therefore, should be made with caution. The results from this study provide parameters for breeders and veterinarians to identify bitches that may require close monitoring for mastitis and metritis.

Public and patient involvement in health data governance (DATAGov): protocol of a people-centred, mixed-methods study on data use and sharing for rare diseases care and research.

INTRODUCTION: International policy imperatives for the public and patient involvement in the governance of health data coexist with conflicting cross-border policies on data sharing. This can challenge the planning and implementation of participatory data governance in healthcare services locally. Engaging with local stakeholders and understanding how their needs, values and preferences for governing health data can be articulated with policies made at the supranational level is crucial. This paper describes a protocol for a project that aims to coproduce a people-centred model for involving patients and the public in decision-making processes about the use and sharing of health data for rare diseases care and research. METHODS AND ANALYSIS: This multidisciplinary project draws on an explanatory sequential mixed-methods study. A hospital-based survey with patients, informal carers, health professionals and technical staff recruited at two reference centres for rare diseases in Portugal will be conducted first. The qualitative study will follow consisting of semi-structured interviews and scenario-based workshops with a subsample of the participant groups recruited at baseline. Quantitative data will be analysed using descriptive and inferential statistics. Inductive and deductive approaches will be combined to analyse the qualitative interviews. Data from scenario-based workshops will be iteratively compared using the constant comparison method to identify cross-cutting themes and categories. ETHICS AND DISSEMINATION: The Ethics Committee for Health from the University Hospital Centre São João/Faculty of Medicine of University of Porto approved the study protocol (Ref. 99/19). Research findings will be disseminated at academic conferences and science promotion events, and through public meetings involving patient representatives, practitioners, policy-makers and students, a project website and peer-reviewed journal publications.

Malunion of an In Utero Metacarpal Fracture in a Standardbred Mare Presenting for Dystocia.

In utero fracture and malunion of long bones is a rare condition in horses. Most foals with in utero fractures are aborted, and the identification of a fetal in utero fracture in a mare with dystocia has not been reported. A 7-year-old multiparous Standardbred mare presented to a referral center for correction of dystocia. Assisted vaginal delivery and controlled vaginal delivery attempts were unsuccessful mainly because of contracted tendons impeding mutation. As the foal was alive, a cesarean section was elected. The foal was delivered but ultimately euthanized because of the congenital abnormalities. Computed tomography of the right forelimb of the foal along with gross examination and histologic evaluation of the right metacarpus revealed the malunion of a previous in utero fracture. While a few cases have been reported of in utero fracture, many of these were in abortuses and not in fetuses at term, making this case a new presentation and potential etiology for dystocia.