Research needs, environmental concerns, and logistical considerations for incorporating livestock grazing into coastal upland habitat management.

Along the Gulf of Mexico (GoM) coast, natural resource managers continually struggle with managing coastal uplands due to front-end costs, prolonged maintenance, and habitat-specific ecological needs. Prescribed fire, mechanical removal, and chemical treatments are common habitat management techniques used to remove invasive species, clear understory, and achieve other management goals. However, rapid development and changing climate exacerbate the difficulty in using these techniques. A potential alternative or complementary technique is using livestock for habitat management (i.e., targeted or controlled grazing). In other regions of the world, using livestock for conservation or restoration of managed lands has shown to be a less intrusive and more financially viable alternative. To better understand the research needs, logistical, and environmental concerns related to using livestock for habitat management in the coastal uplands of the GoM, we developed and distributed a survey to three groups of land users, including natural resource managers, researchers, and livestock producers in the region. Survey results show that over 96% of respondents are interested in using livestock for habitat management, but less than 10% of respondents were aware of any information that could be used to inform grazing practices for coastal upland habitat management along the Gulf of Mexico coast. There were differences among surveyed groups, but generally small-sized cattle breeds and goats were identified as the livestock with the most potential for environmental benefit and ease of containment. General concerns and areas for further investigation were implementation (e.g., which livestock type to use and grazing intensity), logistical considerations (e.g., fencing and rotational frequency), impacts of grazing on water quality, wildlife, vegetation, and livestock nutrition. Survey respondents overwhelmingly (at least 75% of each group) indicated that livestock grazing ideally would not be a standalone management practice and should be used in conjunction with other habitat management techniques such as prescribed burns, mechanical clearing, or chemical treatments. The results of the survey could be used to develop applied research projects and guidance documents that directly address informational needs related to using livestock for habitat management of coastal uplands along the Gulf of Mexico coast.

Systematic review of topical interventions for the management of odour in patients with chronic or malignant fungating wounds.

Chronic wounds adversely affect the quality of life of individuals and odour is a well-recognised associated factor. Odour can affect sleep, well-being, social interactions, diet and potentially wound healing. This systematic review aims to examine the effectiveness of topical interventions in the management of odour associated with chronic and malignant fungating wounds. A systematic review guided by PRISMA recommendations of randomised controlled trials where odour intensity/odour is the primary outcome was undertaken. Inclusion criteria were adults (18 years and over) with chronic venous, arterial, diabetic or pressure ulcers or with malignant fungating wounds where odour has been managed through topical application of pharmacological/non-pharmacological agents. Searches were conducted in CENTRAL, CINAHL, EMBASE, MEDLINE, Scopus, and Web of Science. Eligibility screening, risk of bias assessment and data extraction was completed by authors working independently. Searches retrieved 171 titles and abstracts (157 post de-duplication). Thirteen studies were retained for full text review of which five (n = 137 individuals) examining the following treatments remained: metronidazole (n = 4), silver (n = 1). Meta-analysis was not possible but individual studies suggest improved outcomes (i.e., reduced odour) using metronidazole. Treatment options to manage wound odour are limited and hampered by lack of clinical trials, small sample sizes, and absence of standardised outcomes and consistent measurement. Whereas metronidazole and silver may have a role in controlling wound odour, robust and well-designed interventions with rigorous procedures and standardised odour outcomes are necessary to evaluate their contribution.

Is self always prioritised? Attenuating the ownership self-reference effect in memory.

The current study demonstrates the abolishment of the Ownership Self Reference Effect (OSRE) when elaborate details of a distant other-referent are provided. In a 2 (High versus Low information) × 2 (Self versus Other) experimental design, we tested the capacity for the SRE to be modulated with social saliency. Using a well-established ownership paradigm (Collard et al., 2020; Cunningham et al., 2008; Sparks et al., 2016), when the other was made socially salient (i.e. details and characteristics about the other were provided to the participant prior to encoding), no SRE emerged, such that self-owned and other-owned items were recalled with comparable accuracy. In contrast, when the other was not salient (i.e., no details about them were provided), participants accurately recalled a higher proportion of self-owned items, demonstrating a typical SRE in source memory. The degree of self- or other- referencing was not related to measured variables of closeness, similarity or shared traits with the other. Although the SRE is an established and robust effect, the findings of the current study illustrate critical circumstances in which the self is no longer prioritised above the other. In line with our predictions, we suggest that the self has automatic attributed social salience (e.g. through ownership) and that enhancing social salience by elaborating details of the other, prioritisation can expand to encapsulate an other beyond the self and influence incidental memory.

The feasibility and effectiveness of a novel online mental health literacy course in supporting university student mental health: a pilot study.

BACKGROUND: There is a need for effective universal approaches to promote and support university student mental health that are scalable and sustainable. In this pilot study we assess the feasibility and acceptability of a fully-digitalized, comprehensive mental health literacy course co-created with and tailored to the needs of undergraduate students. We also explore preliminary associations with mental health and positive behaviour change. METHODS: An accredited online mental health literacy course was developed using state-of-the-art pedagogical principles and a reverse mentorship approach. The course was offered as an interdisciplinary undergraduate elective. Students completed an online survey before and after the 12-week course that collected demographic information and assessed mental health knowledge, emotional self-awareness, mental health, stigma, and health-related behaviors using validated measures. Dependent group t-tests were used to compare pre- and post-course levels of knowledge, mental health, sleep quality and substance use. Mental health outcomes of students who completed the course were compared to an age and sex-matched sample of students not enrolled in the course and who completed the same survey measures over the same academic year. Multivariable linear regression was used to examine the effect of course participation on outcomes at follow-up. RESULTS: The course had good uptake and was positively reviewed by participants. Specifically, students found the course engaging, relevant, and applicable, and agreed they would recommend it to their peers. Among course participants there was improvement in mental health knowledge (p < 0.001) and emotional self-awareness (p = 0.02) at course completion. Compared to the matched comparison group, taking the course was associated with reduced alcohol (ß = - 0.41, p = 0.01) and cannabis use (ß = - 0.35, p = 0.03), and improved sleep quality (ß = 1.56, p = 0.09) at the end of the term. CONCLUSIONS: Findings suggest that delivering mental health literacy as an online accredited course may be an acceptable and effective way of promoting university student mental health through improved knowledge, emotional self-awareness, and healthy lifestyle choices. As the course is expanded to larger and more diverse student cohorts we will be able to further examine the short and long-term effectiveness of the course in supporting student mental health and the underlying mechanisms.

Non-Bee Insects as Visitors and Pollinators of Crops: Biology, Ecology, and Management.

Insects other than bees (i.e., non-bees) have been acknowledged as important crop pollinators, but our understanding of which crop plants they visit and how effective they are as crop pollinators is limited. To compare visitation and efficiency of crop-pollinating bees and non-bees at a global scale, we review the literature published from 1950 to 2018 concerning the visitors and pollinators of 105 global food crops that are known to benefit from animal pollinators. Of the 105 animal-pollinated crops, a significant proportion are visited by both bee and non-bee taxa (n = 82; 77%), with a total gross domestic product (GDP) value of US$780.8 billion. For crops with a narrower range of visitors, those that favor non-bees (n = 8) have a value of US$1.2 billion, compared to those that favor bees (n = 15), with a value of US$19.0 billion. Limited pollinator efficiency data were available for one or more taxa in only half of the crops (n = 61; 58%). Among the non-bees, some families were recorded visiting a wide range of crops (>12), including six families of flies (Syrphidae, Calliphoridae, Muscidae, Sarcophagidae, Tachinidae, and Bombyliidae), two beetle families (Coccinelidae and Nitidulidae), ants (Formicidae), wasps (Vespidae), and four families of moths and butterflies (Hesperiidae, Lycaenidae, Nymphalidae, and Pieridae). Among the non-bees, taxa within the dipteran families Syrphidae and Calliphoridae were the most common visitors to the most crops, but this may be an artifact of the limited data available. The diversity of species and life histories in these groups of lesser-known pollinators indicates that diet, larval requirements, and other reproductive needs will require alternative habitat management practices to bees.

Cretaceous origins of the vibrotactile bill-tip organ in birds.

Some probe-foraging birds locate their buried prey by detecting mechanical vibrations in the substrate using a specialized tactile bill-tip organ comprising mechanoreceptors embedded in densely clustered pits in the bone at the tip of their beak. This remarkable sensory modality is known as 'remote touch', and the associated bill-tip organ is found in probe-foraging taxa belonging to both the palaeognathous (in kiwi) and neognathous (in ibises and shorebirds) clades of modern birds. Intriguingly, a structurally similar bill-tip organ is also present in the beaks of extant, non-probing palaeognathous birds (e.g. emu and ostriches) that do not use remote touch. By comparison with our comprehensive sample representing all orders of extant modern birds (Neornithes), we provide evidence that the lithornithids (the most basal known palaeognathous birds which evolved in the Cretaceous period) had the ability to use remote touch. This finding suggests that the occurrence of the vestigial bony bill-tip organ in all modern non-probing palaeognathous birds represents a plesiomorphic condition. Furthermore, our results show that remote-touch probe foraging evolved very early among the Neornithes and it may even have predated the palaeognathous-neognathous divergence. We postulate that the tactile bony bill-tip organ in Neornithes may have originated from other snout tactile specializations of their non-avian theropod ancestors.

The effect of DAFNE education, continuous subcutaneous insulin infusion, or both in a population with type 1 diabetes in Scotland.

AIM: To investigate the effect of DAFNE and continuous subcutaneous insulin infusion in clinical practice. METHODS: Within NHS Lothian, continuous subcutaneous insulin infusion started in 2004 and DAFNE education began in 2006. We extracted anonymized data from the national database for all those aged > 18 years with type 1 diabetes having a Dose Adjustment For Normal Eating course or continuous subcutaneous insulin infusion start date (n = 4617). RESULTS: In total, 956 persons received DAFNE education, and 505 had received an insulin pump, 208 of whom had DAFNE education followed by insulin pump. Mean (SD) HbA1c before DAFNE education was 68 (15) mmol/mol (8.4% [1.4%]) and 66 (13) mmol/mol (8.2% [1.2%]) before continuous subcutaneous insulin infusion. In the year following DAFNE education, the mean fall in within-person HbA1c was 3.8 mmol/mol (95% CI 4.0 to 3.4; 0.3% [0.4% to 0.3%]). Those with the poorest control (HbA1c ≥ 85 mmol/mol [9.9%]) experienced the largest decline (15.7 mmol/mol [1.4%]). Those in the lowest HbA1c band at initiation (< 53 mmol/mmol [7.0%]) experienced a rise. In the year following continuous subcutaneous insulin infusion initiation there was a mean fall in within-person HbA1c of 6.6 mmol/mol (6.8 to 6.4; 0.6% [0.6% to 0.6%]). In those with the poorest control (HbA1c ≥ 85 mmol/mol [9.9%]), the mean fall in HbA1c was 22.2 mmol/mol (23 to 21; 2.0% [2.1% to 1.9%]). Continuous subcutaneous insulin infusion effectiveness was not different with or without DAFNE education. The effects of both interventions were sustained over 5 years. CONCLUSIONS: Both DAFNE education and insulin pump therapy had the greatest effect on HbA1c in those with higher baseline values. There was little difference to attained HbA1c when Dose Adjustment For Normal Eating education was introduced before insulin pump therapy.

High temperatures are associated with substantial reductions in breeding success and offspring quality in an arid-zone bird.

During hot weather, terrestrial animals often seek shaded thermal refugia. However, this can result in missed foraging opportunities, loss of body condition and impaired parental care. We investigated whether such costs could compromise breeding success in a widespread southern African bird: the Southern Yellow-Billed Hornbill Tockus leucomelas. We predicted that hornbills might be especially vulnerable to temperature-dependant reductions in parents' foraging capacity due to extreme asymmetry in sex-specific roles during breeding: females are confined within the nest cavity for most of the nesting period and the burden of provisioning falls solely on the male during this time. We followed 50 hornbill nesting attempts in the Kalahari Desert between 2012 and 2015, collecting data on provisioning rates, adult and nestling body mass, fledging success and size of fledglings. Mean daily maximum air temperatures (Tmax) during nesting attempts ranged from 33.2 to 39.1 °C. The likelihood of successful fledging fell below 50% at mean Tmax > 35.1 °C; a threshold now regularly exceeded at our study site due to recent climate warming. Additionally, offspring fledging following the hottest nesting attempts were > 50% lighter than those fledging following the coolest. Sublethal costs of keeping cool including loss of body condition, production of poor-quality offspring and breeding failure are likely to become issues of serious conservation concern as climate change progresses; even for currently widespread species. Missed-opportunity costs associated with behavioral thermoregulation and direct sublethal costs of temperature exposure should not be overlooked as a potential threat to populations, especially in environments that are already hot.

The costs of keeping cool: behavioural trade-offs between foraging and thermoregulation are associated with significant mass losses in an arid-zone bird.

Avian responses to high environmental temperatures include retreating to cooler microsites and/or increasing rates of evaporative heat dissipation via panting, both of which may affect foraging success. We hypothesized that behavioural trade-offs constrain the maintenance of avian body condition in hot environments, and tested predictions arising from this hypothesis for male Southern Yellow-billed Hornbills (Tockus leucomelas) breeding in the Kalahari Desert. Operative temperatures experienced by the hornbills varied by up to 13 °C among four microsite categories used by foraging males. Lower prey capture rates while panting and reductions associated with the occupancy of off-ground microsites, resulted in sharp declines in foraging efficiency during hot weather. Consequently, male body mass (Mb) gain between sunrise and sunset decreased with increasing daily maximum air temperature (Tmax), from ~ 5% when Tmax < 25 °C to zero when Tmax = 38.4 °C. Overnight Mb loss averaged ~ 4.5% irrespective of Tmax, creating a situation where nett 24-h Mb loss approached 5% on extremely hot days. These findings support the notion that temperature is a major determinant of body condition for arid-zone birds. Moreover, the strong temperature dependence of foraging success and body condition among male hornbills provisioning nests raises the possibility that male behavioural trade-offs translate into equally strong effects of hot weather on female condition and nest success. Our results also reveal how rapid anthropogenic climate change is likely to substantially decrease the probability of arid-zone birds like hornbills being able to successfully provision nests while maintaining their own condition.

Interprofessional education and collaboration: A simulation-based learning experience focused on common and complementary skills in an acute care environment.

The purpose of this mixed-methods study was to explore from the participant's perspective the influence of an interprofessional simulation-based learning experience on understanding the roles and responsibilities of healthcare professionals in the acute care setting, interprofessional collaboration, and communication. Participating students from two professional programs completed the Readiness for Interprofessional Learning Scale (RIPLS) prior to and following the simulation experience to explore the influence of the simulation experience on students' perceptions of readiness to learn together. A Wilcoxon signed rank analysis was performed for each of the four subscales of the RIPLS: shared learning (<.001), teamwork and collaboration (<.001), professional identity (.042), and roles and responsibilities (.001). In addition, participating students were invited to participate in focus group interviews to discuss the effectiveness of the simulation experience. Three key themes were discovered: interprofessional teamwork, discovering roles and responsibilities, and increased confidence in treatment skills. The integration of interprofessional education through a simulation-based learning experience within the nursing and physical therapy professional programs provided a positive experience for the students. Simulation-based learning experiences may provide an opportunity for institutions to collaborate and provide additional engagement with healthcare professions that may not be represented within a single institution.

HbA1c variability is associated with increased mortality and earlier hospital admission in people with Type 1 diabetes.

AIM: Despite evidence of morbidity, no evidence exists on the relationship between HbA1c variability and mortality in Type 1 diabetes. We performed an observational study to investigate whether the association between HbA1c variability and mortality exists in a population of people with Type 1 diabetes. As a secondary outcome, we compared onset of first hospital admission between groups. METHODS: People with Type 1 diabetes were identified for inclusion from the Scottish Care Information - Diabetes data set. This database includes data of all people known to have diabetes who live within Scotland. A survival analysis was carried out over a 47-month period comparing two groups; group 1 with a HbA1c coefficient of variation (CV) above the median CV value, and group 2 with a CV below the median value. Time to death or first admission was also analysed. A Cox proportional hazard model was used to compare time to death, adjusting for appropriate covariables. RESULTS: Some 6048 individuals with Type 1 diabetes were included in the analysis. Median HbA1c CV was 7.9. The hazard ratio (HR) for mortality for those with an HbA1c CV above the median value is 1.5 over 47 months of follow-up (P < 0.001). HR for survival to either the first admission to hospital or death for those with an HbA1c CV above the median value was 1.35 (95% confidence interval 1.25-1.45) over 730 days of follow-up (P < 0.001). CONCLUSION: Our results show that people with greater HbA1c variability have a higher rate of mortality and earlier hospital admission in Type 1 diabetes.

Ureaplasma parvum causes hyperammonemia in a pharmacologically immunocompromised murine model.

A relationship between hyperammonemia and Ureaplasma infection has been shown in lung transplant recipients. We have demonstrated that Ureaplasma urealyticum causes hyperammonemia in a novel immunocompromised murine model. Herein, we determined whether Ureaplasma parvum can do the same. Male C3H mice were given mycophenolate mofetil, tacrolimus, and prednisone for 7 days, and then challenged with U. parvum intratracheally (IT) and/or intraperitoneally (IP), while continuing immunosuppression over 6 days. Plasma ammonia concentrations were determined and compared using Wilcoxon rank-sum tests. Plasma ammonia concentrations of immunosuppressed mice challenged IT/IP with spent broth (median, 188 µmol/L; range, 102-340 µmol/L) were similar to those of normal (median, 226 µmol/L; range, 154-284 µmol/L, p > 0.05), uninfected immunosuppressed (median, 231 µmol/L; range, 122-340 µmol/L, p > 0.05), and U. parvum IT/IP challenged immunocompetent (median, 226 µmol/L; range, 130-330 µmol/L, p > 0.05) mice. Immunosuppressed mice challenged with U. parvum IT/IP (median 343 µmol/L; range 136-1,000 µmol/L) or IP (median 307 µmol/L; range 132-692 µmol/L) had higher plasma ammonia concentrations than those challenged IT/IP with spent broth (p < 0.001). U. parvum can cause hyperammonemia in pharmacologically immunocompromised mice.

Transient suppression of hepatocellular replication in the mouse liver following transduction with recombinant adeno-associated virus.

Recombinant vectors based on adeno-associated virus (AAV) are proving to be powerful tools for genetic manipulation of the liver, for both discovery and therapeutic purposes. The system can be used to deliver transgene cassettes for expression or, alternatively, DNA templates for genome editing via homologous recombination. The replicative state of target cells is known to influence the efficiency of these processes and knowledge of the host-vector interactions involved is required for optimally effective vector deployment. Here we show, for the first time in vivo, that in addition to the known effects of hepatocellular replication on AAV-mediated gene transfer, the vector itself exerts a potent, albeit transient suppressive effect on cell cycle progression that is relieved on a time course that correlates with the known rate of clearance of input single-stranded vector DNA. This finding requires further mechanistic investigation, delineates an excellent model system for such studies and further deepens our insight into the complexity of interactions between AAV vectors and the cell cycle in a clinically promising target tissue.

Does morphology predict trophic position and habitat use of ant species and assemblages?

A functional traits-based theory of organismal communities is critical for understanding the principles underlying community assembly, and predicting responses to environmental change. This is particularly true for terrestrial arthropods, of which only 20% are described. Using epigaeic ant assemblages, we asked: (1) can we use morphological variation among species to predict trophic position or preferred microhabitat; (2) does the strength of morphological associations suggest recent trait divergence; (3) do environmental variables at site scale predict trait sets for whole assemblages? We pitfall-trapped ants from a revegetation chronosequence and measured their morphology, trophic position [using C:N stoichiometry and stable isotope ratios (δ)] and characteristics of microhabitat and macrohabitat. We found strong associations between high trophic position (low C:N and high δ(15)N) in body tissue and morphological traits: predators were larger, had more laterally positioned eyes, more physical protection and tended to be monomorphic. In addition, morphological traits were associated with certain microhabitat features, e.g. smaller heads were associated with the bare ground microhabitat. Trait-microhabitat relationships were more pronounced when phylogenetic adjustments were used, indicating a strong influence of recent trait divergences. At the assemblage level, our fourth corner analysis revealed associations between the prevalence of traits and macrohabitat, although these associations were not the same as those based on microhabitat associations. This study shows direct links between species-level traits and both diet and habitat preference. Trait-based prediction of ecological roles and community structure is thus achievable when integrating stoichiometry, morphology and phylogeny, but scale is an important consideration in such predictions.

[Whipple's disease, when to think about it?]. / Maladie de Whipple: quand y penser?

Whipple's disease is a chronic, systemic, bacterial infection caused by Tropheryma whipplei. Its cardinal symptoms include intermittent and recurrent arthralgia or arthritis together with chronic diarrhoea, abdominal pain and weight loss. It may mimick many chronic inflammatory diseases, and the diagnosis remains a challenge. Salivary and faecal quantitative PCR for T. whipplei should be ensued, if positive, by an upper endoscopy for duodenal biopsies. The treatment consists of a combination of oral doxycycline and hydroxychloroquine for 12 months followed by life-long doxycycline. Whipple's disease, although rare, is an entity that should be considered regularly, as its progression may be fatal if left untreated.

Inhibition controls for qualitative real-time PCR assays: are they necessary for all specimen matrices?

A retrospective analysis of 386,706 specimens representing a variety of matrix types used in qualitative real-time PCR assays determined the overall inhibition rate to be 0.87% when the inhibition control was added preextraction to 5,613 specimens and 0.01% when the inhibition control was added postextraction but preamplification in 381,093 specimens. Inhibition rates of ≤ 1% were found for all specimen matrix types except urine and formalin-fixed, paraffin-embedded tissue.

Massive splenomegaly correlates with malignancy: 180 cases of splenic littoral cell tumors in the world literature.

Littoral cell tumors (LCT) are rare primary splenic neoplasms, unique for their morphologic and immunolabeling features resembling the endothelial littoral cells lining the sinusoids of the red pulp. They include the more common and typically benign littoral cell angioma, as well as the less common, potentially malignant, littoral cell hemangioendothelioma (LCHE) and the aggressive littoral cell angiosarcoma (LCAS). The most common presentation of these neoplasms is splenomegaly, and diagnosis is made histologically following biopsy or resection. To better understand these tumors, a comprehensive, international literature search was performed. Patient and tumor data, including presenting symptoms, comorbid cancers, immunosuppressive states, splenic mass and tumor size were analyzed. Massive splenomegaly (≥ 1500 g) following splenic resection, which correlates with a splenic length of 20 cm preoperatively, was found to be significantly associated with the presence of malignancy in the LCT (P<0.05).

Investigating practice integration of independent prescribing by community pharmacists using normalization process theory: a cross-sectional survey.

BACKGROUND: Independent prescribing (IP) has not been extensively investigated in community pharmacy (CP). Normalization process theory (NPT) constructs help explain how interventions are integrated into practice and include: 'coherence' (understanding), 'cognitive participation' (what promotes engagement), 'collective action' (integration with existing systems), and 'reflexive monitoring' (evaluation). AIM: To use NPT to investigate the integration of pharmacist IP in CP. METHOD: NHS Scotland Pharmacy First Plus (PFP) is a community pharmacy IP service. Questionnaire items were developed using the NPT derived Normalisation MeAsure Development (NoMAD) tool for an online survey of all PFP IP pharmacists. Demographic data were analysed descriptively and scale scores (calculated from item scores for the 4 NPT constructs) were used for inferential analysis. RESULTS: There was a 73% (88/120) response rate. Greater than 90% 'strongly agreed'/'agreed' to NoMAD items relating to most NPT constructs. However, responses to 'collective action' items were diverse with more participants answering 'neither agree nor disagree' or 'disagree'. A statistically significant difference in NPT construct scale scores with significant p-values (ranging from p < 0.001 to p = 0.033) was shown on all the NPT constructs for the variable 'On average, how often do you consult with patients under the PFP service?'. CONCLUSION: This theory-based work offers perspectives on IP integration within CP. Despite its geographic focus this work offers insights relevant to wider contexts on IP integration. It shows 'collective action' focused 'organisation' and 'group process' challenges with a need for further work on staff training, resource availability and utilisation, working relationships, communication and management.

Comparison of gene transfer to the murine liver following intraperitoneal and intraportal delivery of hepatotropic AAV pseudo-serotypes.

Adeno-associated virus (AAV) vectors are highly efficient for liver-targeted gene delivery in murine models and show promise in early phase human clinical trials. This efficiency is capsid-dependent and was only achieved after discovery that the AAV2 vector genome could be trans-encapsidated into the capsids of other AAV serotypes. This confers novel host-vector biology and target tissue tropism. Optimal exploitation of the growing number of AAV vector pseudo-serotypes, however, requires detailed context-dependent characterisation of transduction performance. In this study, we compared the pattern and efficiency of gene delivery to the adult mouse liver following intraportal and intraperitoneal injection of vectors pseudo-serotyped with known hepatotropic capsids from AAV type 7, 8, 9 and rhesus 10. Vectors pseudo-serotyped with these hepatotropic capsids proved relatively efficient irrespective of administration route, with higher transgene expression in males despite equivalent vector genome delivery in females. Transgene expression was predominantly centrilobular in contrast to the AAV2 capsid, which gave a periportal pattern of expression. Most intriguingly, vector genome performance appeared to be delivery route-dependent, consistent with the possibility of in vivo capsid modification. These data not only inform the experimental use of AAV vectors, but also provide insight into novel aspects of host-vector biology requiring further focused analysis.

AAV-encoded OTC activity persisting to adulthood following delivery to newborn spf(ash) mice is insufficient to prevent shRNA-induced hyperammonaemia.

Urea cycle defects presenting in the neonatal period with hyperammonaemia are associated with high morbidity and mortality, and necessitate liver transplantation for long-term management. Gene therapy is therefore an attractive possibility, with vectors based on adeno-associated virus (rAAV) currently showing exciting promise in liver-targeted clinical trials in adults. Successful use of rAAV vectors in infants, however, is more challenging as episomal rAAV genomes will be lost from proliferating hepatocytes during liver growth, leaving stable transgene expression dependent on the subset of vector genomes that undergo genomic integration. To explore this challenge, we exploited the partially ornithine transcarbamylase (OTC)-deficient spf(ash) mouse model and small hairpin RNA-mediated knockdown of residual endogenous OTC enzyme activity in adult mice that had received neonatal treatment with an OTC-encoding rAAV. This leaves mice reliant on vector-encoded OTC activity that has persisted from the newborn period. Despite stable transduction in approximately 8% of hepatocytes and residual vector-encoded OTC activity of up to 33% of wild-type, well above endogenous spf(ash) levels (5-7%), mice were not protected from hyperammonaemia. These data show that the distribution of OTC activity within the liver is critical and that rAAV vector re-delivery after early neonatal treatment is likely to be necessary for stable control of hyperammonaemia into adulthood.