The Toxoplasma gondii F-Box Protein L2 Functions as a Repressor of Stage Specific Gene Expression.

Toxoplasma gondii is a foodborne pathogen that can cause severe and life-threatening infections in fetuses and immunocompromised patients. Felids are its only definitive hosts, and a wide range of animals, including humans, serve as intermediate hosts. When the transmissible bradyzoite stage is orally ingested by felids, they transform into merozoites that expand asexually, ultimately generating millions of gametes for the parasite sexual cycle. However, bradyzoites in intermediate hosts differentiate exclusively to disease-causing tachyzoites, which rapidly disseminate throughout the host. Though tachyzoites are well-studied, the molecular mechanisms governing transitioning between developmental stages are poorly understood. Each parasite stage can be distinguished by a characteristic transcriptional signature, with one signature being repressed during the other stages. Switching between stages require substantial changes in the proteome, which is achieved in part by ubiquitination. F-box proteins mediate protein poly-ubiquitination by recruiting substrates to SKP1, Cullin-1, F-Box protein E3 ubiquitin ligase (SCF-E3) complexes. We have identified an F-box protein named Toxoplasma gondii F-Box Protein L2 (TgFBXL2), which localizes to distinct perinucleolar sites. TgFBXL2 is stably engaged in an SCF-E3 complex that is surprisingly also associated with a COP9 signalosome complex that negatively regulates SCF-E3 function. At the cellular level, TgFBXL2-depleted parasites are severely defective in centrosome replication and daughter cell development. Most remarkable, RNAseq data show that TgFBXL2 conditional depletion induces the expression of stage-specific genes including a large cohort of genes necessary for sexual commitment. Together, these data suggest that TgFBXL2 is a latent guardian of stage specific gene expression in Toxoplasma and poised to remove conflicting proteins in response to an unknown trigger of development.

mRNA-encoded Cas13 treatment of Influenza via site-specific degradation of genomic RNA.

The CRISPR-Cas13 system has been proposed as an alternative treatment of viral infections. However, for this approach to be adopted as an antiviral, it must be optimized until levels of efficacy rival or exceed the performance of conventional approaches. To take steps toward this goal, we evaluated the influenza viral RNA degradation patterns resulting from the binding and enzymatic activity of mRNA-encoded LbuCas13a and two crRNAs from a prior study, targeting PB2 genomic and messenger RNA. We found that the genome targeting guide has the potential for significantly higher potency than originally detected, because degradation of the genomic RNA is not uniform across the PB2 segment, but it is augmented in proximity to the Cas13 binding site. The PB2 genome targeting guide exhibited high levels (>1 log) of RNA degradation when delivered 24 hours post-infection in vitro and maintained that level of degradation over time, with increasing multiplicity of infection (MOI), and across modern influenza H1N1 and H3N2 strains. Chemical modifications to guides with potent LbuCas13a function, resulted in nebulizer delivered efficacy (>1-2 log reduction in viral titer) in a hamster model of influenza (Influenza A/H1N1/California/04/09) infection given prophylactically or as a treatment (post-infection). Maximum efficacy was achieved with two doses, when administered both pre- and post-infection. This work provides evidence that mRNA-encoded Cas13a can effectively mitigate Influenza A infections opening the door to the development of a programmable approach to treating multiple respiratory infections.

Health literacy among fathers and fathers-to-be: a multi-country, cross-sectional survey.

During pregnancy and early fatherhood, men are at higher risk of poor health, exacerbated by low engagement by healthcare services. Yet the transition to fatherhood presents an opportunity for men to improve their health and health behaviours. Health literacy refers to individuals' competence in accessing and applying health information. Poor health literacy is associated with poor health and low help-seeking. The aim of this study was to identify health literacy strengths, needs and profiles among fathers. Men who were expecting a baby ('antenatal') or had become fathers in the past 18 months ('postnatal') were recruited through an international, online paid survey platform. The survey included the nine-scale Health Literacy Questionnaire (HLQ). Of 889 survey respondents (n = 416, 46.5% antenatal; n = 473, 53.5% postnatal), 274 (31.0%) were residing in the USA and 239 (27.0%) in the UK. Relatively higher scores were reported for HLQ scales relating to having sufficient information and finding and understanding this information, as well as social support for health. Relatively lower scores were obtained for scales relating to actively managing one's own health and navigating the health care system. Three scale scores were significantly lower among nulliparous than multiparous men. Seven health literacy profiles were identified. In conclusion, while fathers have some health literacy strengths, they also experience some barriers, particularly first-time fathers. Awareness of diverse health literacy profiles among fathers may assist in developing strategies to strengthen health services' capacity to meet fathers' needs and reduce risks to their health at this critical juncture in families' lives.

What Injured Workers With Complex Claims Look For in Online Communities: Netnographic Analysis.

BACKGROUND: Improved understanding of social constructs around injury may help insurance case managers to understand how best to support people after injury. OBJECTIVE: This study sought to explore what people who sustain work-related injuries may seek from online communities. The study highlights potential opportunities for improved engagement with insurance case management practice. METHODS: An observational netnographic analysis was undertaken on anonymous, publicly available messages posted on Australian message boards. All research data were drawn from anonymous, online communities. A person (author SM) with experience of making a claim through an Australian workers' compensation system and online engagement was involved in study conception, design, and analysis. Data were analyzed using NVivo12 in an iterative, multistage process including coding, journaling, and member checking. A total of 141 people were engaged in discussion across 47 threads housed on 4 Australian forums. RESULTS: In this qualitative study, themes emerged from the data, describing how injured workers use online communities to help make decisions, get support, and solve problems. The key motivators for action and engagement were seeking information, connection, or justice. Establishment of relationships was a key mediator of each of these parameters. CONCLUSIONS: Some work-related injuries may involve medical and medicolegal complexity as well as changed lifestyle and routine during convalescence and recovery. The mechanism used by some injured workers to seek information and problem solve suggests a capacity for self-management and self-care after work-related injury. Netnography provides information on a community that may not regularly engage with research because of the complexity of their situation and their vulnerability.

Gestational Vitamin D and Offspring Bone Measures: Is the Association Independent of Maternal Bone Quality?

Previously we have reported an association between maternal vitamin D and offspring bone as measured by dual-energy X-ray absorptiometry. It is plausible that shared genetics might confound associations between maternal vitamin D in pregnancy and offspring bone measures. We aimed to determine whether such associations are independent of maternal bone quality. Data for this analysis were derived from 168 mother-child pairs who returned at the 11-year follow-up of the vitamin D in pregnancy study. Gestational 25-hydroxyvitamin D [25(OH)D] was assessed by radioimmunoassay in early pregnancy at recruitment (before 16 weeks gestation) and later in pregnancy (28-32 weeks gestation). Bone quality was assessed for mothers and children at the calcaneus using quantitative ultrasound (Achilles InSight, GE). Speed of Sound (SOS), Broadband Ultrasound Attenuation (BUA) and Stiffness Index (SI) were the outcomes of interest. Maternal 25(OH)D in early pregnancy was associated with offspring SOS (ß 1.46 m/s 95% CI 0.12, 2.8). When separated by sex, there was no association between maternal 25(OH)D at recruitment and offspring SI (r = - 0.05, p = 0.68), SOS (r = 0.11, p = 0.34) or BUA (- 0.09, p = 0.43) in girls. In boys, maternal 25(OH)D at recruitment was associated with SI (r = 0.21, p = 0.048), and SOS (r = 0.24, p = 0.03) but not BUA (r = 0.10, p = 0.37). Adjustment for the offspring factors and respective maternal QUS parameter did not attenuate associations between maternal 25(OH)D in early pregnancy with offspring SOS, nor SI. There was no association with BUA. Furthermore, there was no association between maternal 25(OH)D in late pregnancy with any offspring QUS parameter. These prospective data support existing evidence of a positive relationship between maternal 25(OH)D levels during early pregnancy and measures of bone health of offspring in childhood, independent of maternal bone phenotype.

Gestational Folate and Offspring Bone Health; The Vitamin D in Pregnancy Study.

Maternal nutritional intake, such as folate and folic acid supplementation, during pregnancy may affect offspring bone health during childhood. We aimed to determine the associations between maternal dietary and supplementary folate intake and offspring bone health measures, including fracture risk. Data were obtained from 160 of 475 mother-child pairs who had returned for the 11-year follow up of the Vitamin D in Pregnancy Study, an observational cohort study. Incident fractures were ascertained from radiological records and dual X-ray absorptiometry was used to measure bone mineral density and content at 11 years of age. Maternal dietary folate intake during pregnancy was determined by Food Frequency Questionnaire and folate supplementation was determined through self-report. Both measures were undertaken at recruitment (before 16 weeks gestation) and at 28-32 weeks' gestation. Multivariable linear regression models and Cox regression models were used to examine associations. Results are presented as per 1000 µg folate for dietary measures. There were significant associations between maternal folate supplementation in early pregnancy (< 16 weeks gestation) and offspring spine bone mineral content (BMC) (ß = 1.53, 95% CI 0.21, 2.86), spine area (ß = 1.10, 95% CI 0.37, 1.82) and total body less head area (ß = 329.30, 95% CI 3.50, 55.20) at the 11-year follow-up. The association between spine BMC was attenuated after adjustment for bone size (ß = 0.13 95% CI - 0.85, 1.10). There was no association between maternal folate supplementation at 28-32 weeks' or maternal dietary intake at either time point with any offspring bone outcome. These data suggest that folate supplementation in early pregnancy may be associated with offspring bone size, but not other bone measures.

The Association Between Muscle Mass and Strength in Relation to Bone Measures in a Paediatric Population: Sex-Specific Effects.

Post-puberty, bone mass displays clear sex-specific patterns. However, research has suggested that a sexual dimorphism in bone mass is evident in younger children and is likely attributable to differences in lean mass. Thus, we aimed to determine whether the association with both overall muscle mass and/or muscle strength was different between the sexes in a paediatric population. Participants were recruited as part of the Vitamin D in Pregnancy Study, Australia. There were 209/402 (52.3%) children at the 11-year follow-up, and 172 had complete data. Children were assessed for bone mineral content (BMC), bone mineral density (BMD) and lean mass by DXA (Lunar). Handgrip strength (kg) was measured using a dynamometer (JAMAR). Linear regression models were adjusted for height, weight, age and pubertal stage. In adjusted models, including both muscle strength and lean mass, the observed association differed between boys and girls. At the spine in boys, BMC and BMD were associated with muscle strength (ß 0.34 [95%CI 0.09-0.59] and 0.008 [95%CI 0.003-0.014]; respectively) but not total muscle mass. However, muscle mass was associated with BMC and BMD at the total body (less head). In girls, spine BMC and BMD were associated with total lean mass (ß 0.95 [95%CI 0.61-1.3] and ß 0.01 [95%CI 0.005-0.02], respectively), with a similar pattern of association with total body (less head) measures. Muscle mass and strength appear to have sexually dimorphic effects on bone mass in school-aged children. These findings should be replicated in longitudinal studies.

The association between a fracture risk tool and frailty: Geelong Osteoporosis Study.

BACKGROUND: Frailty is characterised by age-related declines in physical, psychological and social functioning. Features of frailty overlap with risk factors for fragility fractures. The aim of this study was to investigate the association between the fracture risk assessment tool (FRAX®) and frailty. METHODS: In cross-sectional analysis, frailty status was determined for participants aged 60-90 yr at 15-year follow-up of the Geelong Osteoporosis Study, using a modified Fried frailty phenotype. Using the FRAX on-line tool, scores for hip and major osteoporotic fracture (MOF) were calculated with and without bone mineral density (BMD). Using the area under Receiver Operating Characteristic (AUROC) curves, and FRAX scores calculated at the baseline visit for these participants, we investigated the association of FRAX and frailty 15 years later. RESULTS: Forty-seven of 303 women (15.5%) and 41 of 282 men (14.5%) were frail at the 15-year visit. There was a gradient of increasing median FRAX scores from robust to frail. For example, for women, median MOF-FRAX without BMD increased from 5.9 for the robust to 7.5 for the pre-frail and 14.0 for the frail (p < 0.001). In secondary analyses, an association was observed between FRAX and frailty over 15 years, with the highest AUROC for women being 0.72 for MOF-FRAX with BMD, and for men, 0.76 hip-FRAX without BMD. CONCLUSION: An association was observed between FRAX and frailty where frail men and women had higher FRAX-scores compared to the other groups. Preliminary data suggest that FRAX, with or without BMD, may be useful in enhancing the information on frailty. Further research using larger datasets will be required to explore this.

Root cause analysis of fall-related hospitalisations among residents of aged care services.

BACKGROUND: Fall-related hospitalisations from residential aged care services (RACS) are distressing for residents and costly to the healthcare system. Strategies to limit hospitalisations include preventing injurious falls and avoiding hospital transfers when falls occur. AIMS: To undertake a root cause analysis (RCA) of fall-related hospitalisations from RACS and identify opportunities for fall prevention and hospital avoidance. METHODS: An aggregated RCA of 47 consecutive fall-related hospitalisations for 40 residents over a 12-month period at six South Australian RACS was undertaken. Comprehensive data were extracted from RACS records including nursing progress notes, medical records, medication charts, hospital summaries and incident reports by a nurse clinical auditor and clinical pharmacist. Root cause identification was performed by the research team. A multidisciplinary expert panel recommended strategies for falls prevention and hospital avoidance. RESULTS: Overall, 55.3% of fall-related hospitalisations were among residents with a history of falls. Among all fall-related hospitalisations, at least one high falls risk medication was administered regularly prior to hospitalisation. Potential root causes of falling included medication initiations and dose changes. Root causes for hospital transfers included need for timely access to subsidised medical services or radiology. Strategies identified for avoiding hospitalisations included pharmacy-generated alerts when medications associated with an increased risk of falls are initiated or changed, multidisciplinary audit and feedback of falls risk medication use and access to subsidised mobile imaging services. CONCLUSIONS: This aggregate RCA identified a range of strategies to address resident and system-level factors to minimise fall-related hospitalisations.

Does 20-min rounding reduce falls in an aged-care setting? A pilot intervention study.

BACKGROUND: This study investigated if implementation of a 20-min rounding intervention can reduce falls in aged care settings. METHODS: Participants (aged 66-99 years) from five aged care facilities were randomly allocated to intervention (n = 20) or control groups (n = 21). The intervention consisted of 20-min rounding observations over a six month period. The number of falls for all residents of each aged care facility was also collected. RESULTS: For participants of the intervention study, there were no differences for number of falls in the intervention compared to the control group (mean(95%CI) control:2.3(0.8-3.7), intervention:4.0(2.5-5.5), p = 0.108). There was a trend for a decreased average number of falls across all aged care sites (mean±SD, 60.4 ± 35.7 falls occurred prior vs. 53.4 ± 37.4 during the intervention, p = 0.056). There were no fall related fractures in the intervention group during the study. CONCLUSIONS: This study suggests that 20-min rounding may decrease falls for all residents of aged care sites.

Health literacy in a population-based sample of Australian women: a cross-sectional profile of the Geelong Osteoporosis Study.

BACKGROUND: The term health literacy refers to the abilities and resources required to find, understand and use health information in managing health. This definition is reflected in the recent development of multidimensional health literacy tools that measure multiple facets of health literacy. The aim of this study was to determine the health literacy profile of a randomly selected, population-based sample of Australian women using a multidimensional tool, the Health Literacy Questionnaire (HLQ). A second aim was to investigate associations between independent HLQ scales, sociodemographic characteristics and lifestyle and anthropometric risk factors for chronic disease. METHODS: We surveyed women involved in the Geelong Osteoporosis Study (GOS), a longitudinal, population-based study. We included demographic data, lifestyle information and anthropometric measures as well as the HLQ. The HLQ has 44 items, scored on either 4- or 5-point scales, within nine conceptually distinct scales. Means for each scale were calculated, and HLQ scales were regressed on educational level and socioeconomic status. Risk factors for chronic disease were investigated using analysis of variance (ANOVA) and calculation of effect sizes. RESULTS: Higher mean scores were seen for the scales 'Feeling understood and supported by healthcare professionals' (mean 3.20, ± SD 0.52) and 'Understanding health information well enough to know what to do' (mean 4.28, ±SD 0.54), and lower mean scores were seen for 'Appraisal of health information' (mean 2.81, ±SD 0.48) and 'Navigating the healthcare system' (mean 4.09, ± SD 0.57). Associations were also seen between lower HLQ scores and poor health behaviours including smoking and being more sedentary, in addition to greater body mass index and waist circumference. Positive gradients were seen between several HLQ scales and education level, as well as SES. For some HLQ scales, these associations were non-linear. CONCLUSIONS: The profile of this population-based cohort of women demonstrated associations between low health literacy and low SES, lower levels of education, increasing age, and anthropometric and lifestyle risk factors for chronic disease. These findings suggest implications of health literacy for health policy makers focusing on improving lifestyle prevention of chronic disease and promoting health equity at a population level.

Maternal Dietary Nutrient Intake During Pregnancy and Offspring Linear Growth and Bone: The Vitamin D in Pregnancy Cohort Study.

Magnesium, phosphorus, zinc, calcium, potassium and protein all play integral roles in maintaining bone health in adults; however, less is known about the importance of these minerals in utero. We aimed to determine associations between maternal dietary consumption of these nutrients during gestation and birth measures in offspring. Of 475 pregnant women recruited from a single antenatal clinic before 16-week gestation (2002-2003) as part of the vitamin D in pregnancy study, 346 with recorded maternal dietary intakes at 28- to 32-week gestation and offspring measures at birth were included. At birth, trained personnel measured the infant's weight, knee-heel length, crown-heel length and head circumference. At age 11, returning offspring underwent assessment of bone mass by dual-energy X-ray absorptiometry (n = 171). Crown-heel length was positively and weakly correlated with maternal intakes of all measured nutrients except calcium, fat and carbohydrate (r = 0.15-0.17; all p ≤ 0.05). The associations with protein, phosphorus and potassium were not attenuated after adjustment for maternal and offspring characteristics. No sustained associations were seen with other birth measures. Further, associations with some nutrients persisted with offspring height at age 11 years. Offspring bone area was associated with maternal diet, but no other measure of bone mass at age 11. After adjustment for height, associations were not significant. These data highlight that whilst some nutritional factors during pregnancy are associated with offspring linear growth in utero and childhood, this does not necessarily translate into an effect on offspring bone measures in childhood.

Lived experiences of parents caring for a child with a life-limiting condition in Australia: A qualitative study.

BACKGROUND: Experiential studies in paediatric palliative care are needed to enable an ongoing international agenda which supports the development of responsive family supports. AIM: To provide an in-depth exploration of the prevalent lived experiences of parents who are currently providing care for a child with a life-limiting condition in Australia. DESIGN: Cross-sectional, prospective, qualitative study guided by an advisory group and reported according to the consolidated criteria for reporting qualitative studies. Transcripts were subjected to a thematic analysis, underpinned by an interpretative phenomenological framework. SETTING/PARTICIPANTS: Purposively sampled parents (n = 14) recruited from a statewide paediatric hospice who self-identified as a 'primary caregiver' for one or more children and/or adolescents (⩽18 years) with a life-limiting condition. RESULTS: Four key themes represented the prevalent experiences of parents: (1) trapped inside the house, (2) the protector, (3) living with the shadow and (4) travelling a different pathway. They describe parents' physical and social isolation, exclusion from the workforce, pervasive grief and associated impacts to their health and well-being. Limited professional and diminished social supports resulted in full ownership of care responsibility. Yet, parents embraced their role as 'protector', reporting acquired meaning and purpose. CONCLUSION: This study builds upon the growing body of evidence available in paediatric palliative care internationally. The key themes highlight the substantial demand for both physical and emotional support beyond what is currently offered and call for the implementation of carefully planned support services and other societal initiatives which seek to alleviate the broad health impacts to caregivers.

Provocative intraocular pressure challenge preferentially decreases venous oxygen saturation despite no reduction in blood flow.

PURPOSE: Ocular disease can both alter the retina's oxygen requirements, and decrease its ability to cope with changes in metabolic demand. We examined the influence of a moderate intraocular pressure (IOP) elevation on three outcome measures: arterial and venous oxygen saturation, blood flow, and the pattern electroretinogram (PERG). METHODS: We increased IOP to ˜30 mmHg in 23 healthy participants (22-39 years) using a mechanical probe applied to the eyelid, thereby lowering ocular perfusion pressure (OPP) by ~30%. The Oxymap retinal oximeter was used to measure oxygen saturation for arteries and veins. Blood flow, volume and velocity were measured using the Heidelberg retinal flowmeter and steady-state PERG waveforms (8.34 Hz) were recorded bilaterally (200 sweeps). For each outcome measure, data was obtained three times: at baseline, 1 min into sustained IOP elevation, and 1 min after the probe was removed. RESULTS: During IOP elevation, changes in oxygen saturation of retinal arteries failed to reach statistical significance [F(1,30) = 3.69, p = 0.05], whereas venous oxygen saturation was significantly reduced [F(1,21) = 27.43, p < 0.01]. Blood flow increased slightly [F(2,40) = 6.28, p < 0.0001], PERG amplitude significantly reduced [F(2,44) = 24.24, p < 0.0001] and PERG phase was significantly delayed [F(2,44) = 17.00, p < 0.0001]. Contralateral eyes were unchanged. OPP reduction correlated little with PERG amplitude, PERG phase or venous oxygen saturation. CONCLUSIONS: Mild, acute IOP elevation increases arterio-venous oxygen saturation differences primarily through lowering venous oxygen saturation, suggesting increased oxygen consumption by healthy neurons when physiologically stressed.

Does the Swedish Interactive Threshold Algorithm (SITA) accurately map visual field loss attributed to vigabatrin?

BACKGROUND: Vigabatrin (VGB) is an anti-epileptic medication which has been linked to peripheral constriction of the visual field. Documenting the natural history associated with continued VGB exposure is important when making decisions about the risk and benefits associated with the treatment. Due to its speed the Swedish Interactive Threshold Algorithm (SITA) has become the algorithm of choice when carrying out Full Threshold automated static perimetry. SITA uses prior distributions of normal and glaucomatous visual field behaviour to estimate threshold sensitivity. As the abnormal model is based on glaucomatous behaviour this algorithm has not been validated for VGB recipients. We aim to assess the clinical utility of the SITA algorithm for accurately mapping VGB attributed field loss. METHODS: The sample comprised one randomly selected eye of 16 patients diagnosed with epilepsy, exposed to VGB therapy. A clinical diagnosis of VGB attributed visual field loss was documented in 44% of the group. The mean age was 39.3 years ± 14.5 years and the mean deviation was -4.76 dB ±4.34 dB. Each patient was examined with the Full Threshold, SITA Standard and SITA Fast algorithm. RESULTS: SITA Standard was on average approximately twice as fast (7.6 minutes) and SITA Fast approximately 3 times as fast (4.7 minutes) as examinations completed using the Full Threshold algorithm (15.8 minutes). In the clinical environment, the visual field outcome with both SITA algorithms was equivalent to visual field examination using the Full Threshold algorithm in terms of visual inspection of the grey scale plots , defect area and defect severity. CONCLUSIONS: Our research shows that both SITA algorithms are able to accurately map visual field loss attributed to VGB. As patients diagnosed with epilepsy are often vulnerable to fatigue, the time saving offered by SITA Fast means that this algorithm has a significant advantage for use with VGB recipients.

Test-retest reliability of retinal oxygen saturation measurement.

PURPOSE: To determine intrasession and intersession repeatability of retinal vessel oxygen saturation from the Oxymap Retinal Oximeter using a whole image-based analysis technique and so determine optimal analysis parameters to reduce variability. METHODS: Ten fundus oximetry images were acquired through dilated pupils from 18 healthy participants (aged 22 to 38) using the Oxymap Retinal Oximeter T1. A further 10 images were obtained 1 to 2 weeks later from each individual. Analysis was undertaken for subsets of images to determine the number of images needed to return a stable coefficient of variation (CoV). Intrasession and intersession variability were quantified by evaluating the CoV and establishing the 95% limits of agreement using Bland and Altman analysis. Retinal oxygenation was derived from the distribution of oxygenation values from all vessels of a given width in an image or set of images, as described by Paul et al. in 2013. RESULTS: Grouped in 10-µm-wide bins, oxygen saturation varied significantly for both arteries and veins (p < 0.01). Between 110 and 150 µm, arteries had the least variability between individuals, with average CoVs less than 5% whose confidence intervals did not overlap with the greater than 10% average CoVs for veins across the same range. Bland and Altman analysis showed that there was no bias within or between recording sessions and that the 95% limits of agreement were generally lower in arteries. CONCLUSIONS: Retinal vessel oxygen saturation measurements show variability within and between clinical sessions when the whole image is used, which we believe more accurately reflects the true variability in Oxymap images than previous studies on select image segments. Averaging data from vessels 100 to 150 µm in width may help to minimize such variability.

Probenecid Inhibits Influenza A(H5N1) and A(H7N9) Viruses In Vitro and in Mice.

Avian influenza (AI) viruses cause infection in birds and humans. Several H5N1 and H7N9 variants are highly pathogenic avian influenza (HPAI) viruses. H5N1 is a highly infectious bird virus infecting primarily poultry, but unlike other AIs, H5N1 also infects mammals and transmits to humans with a case fatality rate above 40%. Similarly, H7N9 can infect humans, with a case fatality rate of over 40%. Since 1996, there have been several HPAI outbreaks affecting humans, emphasizing the need for safe and effective antivirals. We show that probenecid potently inhibits H5N1 and H7N9 replication in prophylactically or therapeutically treated A549 cells and normal human broncho-epithelial (NHBE) cells, and H5N1 replication in VeroE6 cells and mice.

"I always felt like I wasn't supposed to be there". An international qualitative study of fathers' engagement in family healthcare during transition to fatherhood.

OBJECTIVE: Engagement of fathers in family health services confers benefits for the health and wellbeing of the whole family. The childbirth continuum is traditionally considered a feminine event, however, commensurate with the changing paradigm of gender equity in family healthcare worldwide, the role of fathers is in transformation. The aim of the study is to explore father's perceptions and experiences of healthcare engagement during pregnancy and early infant care. DESIGN: Qualitative free-text questions were embedded in a large multi-country, cross-sectional survey, to explored fathers' attendance, participation, and experience of health care during appointments with their pregnant partner and/or baby. SETTING AND PARTICIPANTS: Expectant and new fathers were recruited through Prolific®, an international paid online survey platform. FINDINGS: Qualitative responses (n=889) were provided by fathers from 28 countries, with experiences of a range of contexts and models of care; 46.8% of whose partners were pregnant and 53.2% had given birth since 2020. The findings suggest that although most fathers wanted to attend and participate in maternity and early parenting-related healthcare, multiple barriers were identified at the individual father, organisational context, and societal levels. Fathers reported negative social factors such as gender bias and restrictive gender norms as barriers to their healthcare engagement. In contrast, factors that enabled fathers to overcome barriers included the fathers' feelings of confidence in their partner's autonomy and decision-making skills, trusted professional relationships with clinicians, and clinicians with good interpersonal skills. KEY CONCLUSIONS: Multiple barriers restrict the participation of fathers in healthcare for childbearing and early parenting. Knowledge of these barriers can inform healthcare redesign to include more successful engagement strategies for fathers, to benefit fathers, mothers, and infants alike. IMPLICATIONS FOR PRACTICE: Health professionals consulting with the mother, father and infant triad are ideally placed to address the healthcare needs of both parents. Early engagement of fathers in family health care by use of inclusive interpersonal skills and the development of a trusted relationship has potential to improve paternal mental health, and may be associated with benefits for the health, wellbeing and safety of the whole family.

Gestational vitamin D and offspring fracture risk: do associations persist into mid adolescence?

BACKGROUND: Previous studies report that maternal vitamin D exposure during pregnancy is associated with offspring later-life bone health. A study in the Vitamin D in Pregnancy (VIP) cohort reported sexually dimorphic effects of maternal 25-hydroxyvitamin-D (25(OH)D) and offspring fracture profiles at 10 years of age. We, therefore, aimed to determine associations between maternal 25(OH)D status and offspring fracture risk at 16 years of age in this cohort. METHODS: In total, 475 mother-child pairs were recruited to the VIP study in southeastern Australia. Maternal serum samples were obtained at recruitment (<16 weeks' gestation) and/or 28-32 weeks' gestation and analysed for 25(OH)D. Radiologically-confirmed incident fractures in children were ascertained from date of birth (2002-2004) until July 16, 2019. Cox proportional hazard models were used to determine associations between maternal 25(OH)D and childhood fracture risk, and final models included maternal age at recruitment, offspring sex, birth weight, gestation length and season of 25(OH)D sample. RESULTS: Data were available for 400 children (mean age 16.1 years). There were 122 (30.5%) children who sustained at least one fracture. Higher maternal 25(OH)D (per 10 nmol/L) in early gestation was associated with a decreased fracture risk in boys (HR 0.87; 95% CI: 0.77, 0.99); the pattern was reversed in girls (HR 1.10; 95% CI 1.00, 1.22). At late gestation, higher maternal 25(OH)D was associated with an increased fracture risk in girls (HR 1.14; 95% CI: 1.04, 1.24). CONCLUSIONS: While our findings must be interpreted within the constraints of our limitations, we report that the contradictory risk profiles observed at early childhood in this cohort remain in adolescence.

Non-invasive detection of microvascular changes in a paediatric and adolescent population with type 1 diabetes: a pilot cross-sectional study.

BACKGROUND: The detection of microvascular damage in type 1 diabetes is difficult and traditional investigations do not detect changes until they are well established. The purpose of this study was to investigate the combined ability of nailfold capillaroscopy, laser Doppler flowmetry, retinal vessel analysis and 24-hr ambulatory blood pressure monitoring to detect early microvascular changes in a paediatric and adolescent population with type 1 diabetes. METHODS: Patients aged between 8 - 18 years with type I diabetes and no other autoimmune conditions were studied. The participants underwent the above cardiac and vascular investigations in a single three-hour session. Standard parameters including HbA1c were also investigated. Associations between all parameters were described by correlation analysis. Fisher's exact and t-tests determined the association with clinical findings. RESULTS: 26 participants were recruited. The mean HbA1c was 8.1% (SD ± 1.1) with a mean duration of type 1 diabetes of 7.9 years (SD ± 3.4). Three participants had microalbuminuria and one had early signs of retinopathy. Participants with microvascular complications had more avascular areas on nailfold capillaroscopy (p = 0.03). Recent HbA1c was positively associated with the number of nailfold microhaemorrhages (p = 0.03) Decreased baseline perfusion by laser Doppler flowmetry was associated with increased capillary density (p = 0.001) and an increased number of microaneurysms (p = 0.04) on nailfold capillaroscopy. CONCLUSIONS: This pilot study has shown that in children and adolescents with established type 1 diabetes, abnormal microvasculature can be detected by these investigations. These markers were also positively associated with evidence of suboptimal diabetes control as assessed by HbA1c. Further research will be necessary to determine the practical role of these investigations in the management and progress of the complications of type 1 diabetes. TRIAL REGISTRATION: Clinical Trial number NCT01279928, ClinicalTrials.gov.