Improving Physiological Relevance of Cell Culture: The Possibilities, Considerations and Future Directions of the Ex Vivo Co-Culture Model

In vitro models provide an important platform for the investigation of cellular growth and atrophy to inform, or extend mechanistic insights from, logistically challenging in vivo trials. While these models allow for the identification of candidate mechanistic pathways, many models involve supraphysiological dosages, non-physiological conditions, or experimental changes relating to individual proteins or receptors, all of which limit translation to human trials. To overcome these drawbacks, the use of ex vivo human plasma and serum has been used in cellular models to investigate changes in myotube hypertrophy, cellular protein synthesis, anabolic and catabolic markers in response to differing age, disease states, and nutrient status. However, there are currently no concurrent guidelines outlining the optimal methodology for this model. This review discusses the key methodological considerations surrounding the use of ex vivoplasma and serum, with a focus in application to skeletal muscle cell lines (i.e., C2C12, L6 and LHCN-M2) and human primary skeletal muscle cells (HSMC) as a means to investigate molecular signaling in models of atrophy and hypertrophy, alongside future directions

Lipid production through the single-step microwave hydrolysis of macroalgae using the oleaginous yeast Metschnikowia pulcherrima

Macroalgae (seaweeds) represent an emerging resource for food and the production of commodity and specialty chemicals. In this study, a single-step microwave process was used to depolymerise a range of macroalgae native to the United Kingdom, producing a growth medium suitable for microbial fermentation. The medium contained a range of mono- and polysaccharides as well as macro- and micronutrients that could be metabolised by the oleaginous yeast Metschnikowia pulcherrima. Among twelve macroalgae species, the brown seaweeds exhibited the highest fermentation potential, especially the kelp Saccharina latissima. Applying a portfolio of ten native M. pulcherrima strains, yeast growth kinetics, as well as production of lipids and 2-phenylethanol were examined, with productivity and growth rate being strain dependent. On the 2 L scale, 6.9 g L−1 yeast biomass – a yield of 0.14 g g−1 with respect to the supplied macroalgae – containing 37.2% (w/w) lipid was achieved through utilisation of the proteins, mono- and polysaccharides from S. latissima, with no additional enzymes. In addition, the yeast degraded a range of fermentation inhibitors released upon microwave processing at high temperatures and long holding times. As macroalgae can be cultured to food grade, this system offers a novel, potentially low-cost route to edible microbial oils as well as a renewable feedstock for oleochemicals

Tuning in to Toddlers: Research Protocol and Recruitment for Evaluation of an Emotion Socialization Program for Parents of Toddlers

Background: Parenting a toddler is a challenging experience for many parents with times of emotional dysregulation in both parent and child. Parenting interventions may be useful for parents to improve their ability to regulate emotions and respond to children’s emotions in a way that assists the child to understand and regulate emotions (emotion competence). Tuning in to Toddlers (TOTS) is a new parenting program that aims to improve parents’ emotion regulation, emotional responsiveness, and emotion coaching (aspects of emotion socialization) to promote optimal emotional development in toddlers, and prevent social and behavioral difficulties. This paper outlines the rationale, methodology, intervention, and recruitment used in a trial to establish program efficacy.Methods/Design: Parents of toddlers aged 18–36 months old were recruited through child care centers (CC) and maternal child health (MCH) centers in Melbourne, Australia and were allocated to either intervention or a 15-month wait-list control condition in a cluster-randomized controlled design. Inclusion criteria were a child in the age range at baseline attending one of the CC or MCH centers. Exclusion criteria were if the parent/carer had insufficient English to attend the intervention and complete measures. Parents in the intervention condition participated in the 6-session group TOTS program delivered by two facilitators using a structured manual and measures of program fidelity and acceptability. Participants in the wait-list control condition received the intervention after a 15-month waiting period. Participants completed measures at baseline, post-intervention (intervention participants only) and 15-month follow-up. Primary outcome measures included parent emotion socialization (parent-report and observed). Secondary outcomes included parent-reported parent functioning (emotion regulation and mental health), toddler social, emotional and behavioral functioning, and parent and toddler systemic cortisol stress (using hair samples). The study was designed to comply with the CONSORT statement and intervention reporting outlined using TIDieR.Results: Three hundred and six parents were recruited and completed baseline parent questionnaires, with a further 234 completing parent–child observation assessments, 235 parent cortisol, and 198 child cortisol.Discussion: This paper is a methodological description of the TOTS randomized controlled trial evaluation protocol. It outlines some of the challenges in recruiting parents of toddlers to parenting programs.Clinical Trial Registration:www.ClinicalTrials.gov, identifier ACTRN12615000 962538

IL-23 plays a key role in Helicobacter hepaticus–induced T cell–dependent colitis

Inflammatory bowel disease (IBD) is a chronic inflammatory disorder of the gastrointestinal tract that is caused in part by a dysregulated immune response to the intestinal flora. The common interleukin (IL)-12/IL-23p40 subunit is thought to be critical for the pathogenesis of IBD. We have analyzed the role of IL-12 versus IL-23 in two models of Helicobacter hepaticus–triggered T cell–dependent colitis, one involving anti–IL-10R monoclonal antibody treatment of infected T cell–sufficient hosts, and the other involving CD4+ T cell transfer into infected Rag−/− recipients. Our data demonstrate that IL-23 and not IL-12 is essential for the development of maximal intestinal disease. Although IL-23 has been implicated in the differentiation of IL-17–producing CD4+ T cells that alone are sufficient to induce autoimmune tissue reactivity, our results instead support a model in which IL-23 drives both interferon γ and IL-17 responses that together synergize to trigger severe intestinal inflammation

Telephone advice lines for adults with advanced illness and their family carers: a qualitative analysis and novel practical framework

BackgroundTelephone advice lines have been recommended internationally to support around-the-clock care for people living at home with advanced illness. While they undoubtedly support care, there is little evidence about what elements are needed for success. A national picture is needed to understand, improve and standardise service delivery/care. Aim To explore telephone advice lines for people living at home with advanced illness across the four UK nations, and to construct a practical framework to improve services.DesignA cross-national evaluation of telephone advice lines using structured qualitative interviews. A patient and public involvement workshop was conducted to refine the framework.Setting/participantsProfessionals with responsibilities for how palliative care services are delivered and/or funded at a local or regional level, were purposively sampled. ResultsSeventy-one interviews were conducted, covering 60 geographical areas. Five themes were identified. Availability: Ten advice line models were described. Variation led to confusion about who to call and when. Accessibility, awareness and promotion: It was assumed that patients/carers know who to call out-of-hours, but often they did not. Practicalities: Call handlers skills/expertise varied, which influenced how calls were managed. Possible responses ranged from signposting to organising home visits. Integration/continuity of care: Integration between care providers was limited by electronic medical records access/information sharing. Service structure/commissioning: Sustained funding was often an issue for charitably funded organisations.ConclusionsOur novel evidence-based practical framework could be transformative for service design/delivery, as it presents key considerations relating to the various elements of advice lines that may impact on the patient/carer experience

Self-reported data: a major tool to assess compliance with anti-malarial combination therapy among children in Senegal

Background: Although there are many methods available for measuring compliance, there is no formal gold standard. Different techniques used to measure compliance were compared among children treated by the anti-malarial amodiaquine/sulphadoxine-pyrimethamine (AQ/SP) combination therapy, in use in Senegal between 2004 and 2006. Methods: The study was carried out in 2004, in five health centres located in the Thies region (Senegal). Children who had AQ/SP prescribed for three and one day respectively at the health centre were recruited. The day following the theoretical last intake of AQ, venous blood, and urine samples were collected for anti-malarial drugs dosage. Caregivers and children above five years were interviewed concerning children's drug intake. Results: Among the children, 64.7% adhered to 80% of the prescribed dose and only 37.7% were strict full adherent to the prescription. There was 72.7% agreement between self-reported data and blood drug dosage for amodiaquine treatment. Concerning SP, results found that blood dosages were 91.4% concordant with urine tests and 90% with self-reported data based on questionnaires. Conclusion: Self-reported data could provide useful quantitative information on drug intake and administration. Under strict methodological conditions this method, easy to implement, can be used to describe patients' behaviors and their use of new anti-malarial treatment. Self-reported data is a major tool for assessing compliance in resource poor countries. Blood and urine drug dosages provide qualitative results that confirm any drug intake. Urine assays for SP could be useful to obtain public health data, for example on chemoprophylaxis among pregnant women

Violence against children and intimate partner violence against women: overlap and common contributing factors among caregiver-adolescent dyads.

BACKGROUND: Intimate partner violence against women (IPV) and violence against children (VAC) are both global epidemics with long-term health consequences. The vast majority of research to date focuses on either IPV or VAC, however the intersections between these types of violence are a growing area of global attention. A significant need exists for empirical research on the overlap of IPV and VAC, especially in contexts with particularly high rates of both types of violence. METHODS: This exploratory study includes secondary analysis of data from a cluster randomized controlled trial in Ugandan schools. Using baseline reports from a random sample of early adolescents attending school and their caregivers, this study uses a probability sample across all eligible schools of adolescent-caregiver dyads (n = 535). We categorized adolescent-caregiver dyads into four groups: those reporting VAC 'only', IPV 'only', both VAC and IPV, or 'no violence'. Two separate multinomial logistic regression models for male and female caregivers explored adolescent and caregiver characteristics associated with the VAC 'only', the IPV 'only', or the both VAC and IPV dyads, each compared to the 'no violence' dyad. RESULTS: One third of dyads reported both IPV and VAC and nearly 75% of dyads reported VAC or IPV. Dyads reporting IPV were more likely to also report VAC. Common contributing factors for female caregiver-adolescent dyads with both VAC and IPV include lower SES, less caregiver education, higher caregiver mental distress, more frequent caregiver alcohol use, and caregivers who report less emotional attachment to their intimate partner. Male caregiver-adolescent dyads with both VAC and IPV included caregivers with less emotional attachment to their intimate partner and more attitudes accepting VAC. CONCLUSIONS: Findings reveal a significant overlap of IPV and VAC and the importance for violence prevention and response programming to consider coordinated or integrated programming. Unique results for female and male caregivers highlight the importance of a gendered approach to addressing IPV and VAC intersections. TRIAL REGISTRATION: The trial was registered at clinicaltrials.gov, NCT01678846, on September 5, 2012

COVID-19 Vaccination of Individuals with Down Syndrome—Data from the Trisomy 21 Research Society Survey on Safety, Efficacy, and Factors Associated with the Decision to Be Vaccinated

Individuals with Down syndrome (DS) are among the groups with the highest risk for severe COVID-19. Better understanding of the efficacy and risks of COVID-19 vaccines for individuals with DS may help improve uptake of vaccination. The T21RS COVID-19 Initiative launched an international survey to obtain information on safety and efficacy of COVID-19 vaccines for individuals with DS. De-identified survey data collected between March and December 2021 were analyzed. Of 2172 individuals with DS, 1973 (91%) had received at least one vaccine dose (57% BNT162b2), 107 (5%) were unvaccinated by choice, and 92 (4%) were unvaccinated for other reasons. Most participants had either no side effects (54%) or mild ones such as pain at the injection site (29%), fatigue (12%), and fever (7%). Severe side effects occurred in <0.5% of participants. About 1% of the vaccinated individuals with DS contracted COVID-19 after vaccination, and all recovered. Individuals with DS who were unvaccinated by choice were more likely to be younger, previously recovered from COVID-19, and also unvaccinated against other recommended vaccines. COVID-19 vaccines have been shown to be safe for individuals with DS and effective in terms of resulting in minimal breakthrough infections and milder disease outcomes among fully vaccinated individuals with DS