Densities for various Tetramethylammonium hydroxide (TMAH) solutions at different temperatures have been determined based on Archimedes’ principle using a hydrostatic balance and a vitreous body with a defined volume. Due to the toxicity of the compound, these data are important especially in the field of wet processing techniques for the automated dosage of TMAH. Specifications Table•Data for the temperature/density/concentration correlation do not exist.•These data are important especially in wet processing applications i.e. for the calibration of concentration measuring units and are therefore important for industry and science.•Chemical engineers will benefit from these data especially for the optimization of wet processing applications.1Tetramethylammonium hydroxide  is a strong basic compound that finds application in both scientific and industrial areas. For example, it is used as a reagent in the thermochemolysis of various biopolymers like lignin 23Chemicals and reagents were purchased from Carl Roth or Merck and were used without further purification. The dilution series was prepared using deionized water and graduated flasks. The concentrations have been confirmed by titration against 0.5 M HCl on test basis.For density measurements the solutions have been transferred to Erlenmeyer flasks which were then equipped with a thermometer plug for direct temperature measurements in the solutions. These then have been placed in a water filled memmert thermostat type W200 and heated to the corresponding temperatures. Only for the runs at 15°C a MaxQ4000 thermostat from Thermo Scientific was used in order to chill the probes below room temperature. The density has been determined for each solution at each temperature three times by using a hydrostatic balance (Mettler PM2000) based on the Archimedes’ principle using a vitreous body with a calibrated volume of 10.005 ml.No experiments on human participants or animals were carried out during the implementation of these experiments, nor were any human or animal rights violated.Josef Diebold: Data curation, Writing – original draft preparation. Yasmin Weiß: Investigation, Validation; Stefan Zürcher: Resources, Methodology; Magnus S. Schmidt: Conceptualization, Supervision, Writing – review & editing.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


The ability to use bio‐inspired building blocks in the assembly of novel supramolecular frameworks is at the forefront of an exciting research field. Herein, we present the first polyproline helix to self‐assemble into a reversibly porous, crystalline, supramolecular peptide framework (SPF). This framework is assembled from a short oligoproline, adopting the polyproline II conformation, driven by hydrogen‐bonding and dispersion interactions. Thermal activation, guest‐induced dynamic porosity and enantioselective guest inclusion have been demonstrated for this novel system. The principles of the self‐assembly associated with this SPF will be used as a blueprint allowing for the further development of helical peptide linkers in the rational design of SPFs and metal‐peptide frameworks. The ability to use bio‐inspired building‐blocks in the assembly of novel supramolecular frameworks is at the forefront of an exciting research field. Herein, we present the first polyproline helix to self‐assemble into a reversibly porous, crystalline, supramolecular peptide framework (SPF). Thermal activation, guest‐induced dynamic porosity and enantioselective guest inclusion have been demonstrated for this novel system. This interconversion can be controlled as a function of the environment it is exposed to  and can be monitored by circular dichroism (CD) spectroscopy. on an Fmoc‐associated pair . Subsequently, simultaneous thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) of PP4‐SPF showed a crystalline melting point with an endothermic peak .Due to the porosity of the framework, it has the potential for “activation” by removal of the solvent from the pores, thereby allowing the introduction of other guests into the activated pores. To investigate the ability for thermal release of ethanol from the framework channels, Powder X‐ray diffraction (PD‐XRD) analyses revealed a change in phase upon de‐solvation, with significant differences to the original sample . To rule out the possibility of partial recrystallization causing the phase change, a sample of PP4‐SPFact was soaked in hexane and showed no change in phase on the powder pattern, as such hexane was used as the diluent for other guest molecules. The framework was soaked in a 5 % solution of various guests . After soaking the SPF was washed thoroughly with fresh hexane to remove excess guest. PDXRD analysis was then carried out to determine whether a change in phase had occurred  for each guest (Supporting Information 13.4). The mol % of guest retained within the pores was analysed by 1H NMR, after dissolving the framework in [D4]methanol .Due to the framework exhibiting dynamic porosity, attempts were made to see if this behaviour would apply with other guests. PP1H NMR data showed significantly higher proportions of hexane than for the SPF soaked alone in hexane , which suggests that once porosity is recovered hexane is adsorbed into the framework. However, the proportion of guest alters significantly , favouring adsorption of guest molecules over the hexane diluent (Supporting Information 13).All the guests analysed  1‐phenylethanol, and iodine), elicited a change in phase from the de‐solvated structure, after analysis of the solid SPF after soaking, suggesting they all act to reinflate the structure (Supporting Information 13.4). However, differences in the powder patterns of the samples suggests either a degree adaptation of the framework to accommodate different guests when compared to the original ethanol guest or ordering of the new guest molecules resulting in new diffraction peaks. Although, differences in peak intensities and loss of some low angle peaks are likely attributable to both loss of crystal quality through the processes of activation and guest inclusion. It should be noted that the activated material loses crystallinity when mechanically ground , and so the measured diffraction patterns for the (unground) materials also exhibit some preferred orientation effects. Analysis of the 4‐SPF ‐1‐phenylethanol)  1‐phenylethanol (PhEtOH) within the framework , chiral HPLC was utilized to investigate whether any enantioselectivity was exhibited (Supporting Information 13.5). These results showed a clear shift in the enantiomeric ratio from the reference racemic (±) PhEtOH after exposure to PP4‐SPF@I2 was performed which showed that the SPF readopted the initial solvated structural geometry and was able to adsorb iodine with a chemical occupancy for I2 of ≈12 % , thus demonstrating the potential of the PP4‐SPF to perform thermally responsive guest release.Activated single crystals were then exposed to a solution of iodine in hexane to allow visualization of a guest within the reinflated pores through single‐crystal diffraction, even at lower guest loadings, due to the high electron density of iodine. A colour change was evident with the crystals taking on a yellow‐orange hue. SC‐XRD analysis of PP% Figure . To inve4‐SPF) using the resilience and rigidity of the polyproline II helix in a short oligoproline is reported. This SPF shows remarkable reversible porosity and the ability to reversibly host chemical guests in its channels with exhibited enantioselectivity of this adsorption process. The self‐assembling principles associated with this SPF can be used as a blueprint in the synthesis of novel helical SPFs and have exciting potential for the rational design of functionalized cavities, offering an exceptional level of positional control of easily incorporated new functionalities. These materials have clear applications in chemical separation and potential chiral catalysis.In conclusion, the first supramolecular peptide framework formed by the self‐assembly of a polyproline helix  should be addressed to the authors.Supporting InformationClick here for additional data file.


Targeted ultraviolet (UV) phototherapy has been used in the management of a wide variety of dermatological clinical conditions including moderate to severe psoriasis unresponsive to topical therapies, vitiligo, severe atopic dermatitis and lymphoproliferative disorders. To date there are no uniform, standardised guidelines for the selection and decontamination process for UV personal protective equipment (PPE) and facial shields used in phototherapy. In the current climate, Coronavirus 2019 (COVID‐19) pandemic, standards regarding all decontamination and disinfection processes are under significant scrutiny. In terms of the UV‐PPE and facial shields used in phototherapy, careful disinfection procedures need to be implemented to ensure that the decontamination practice is effective enough to neutralise the virulent virus whilst maintaining maximal protection to the user from UV‐rays and safeguard the equipment from damage during the cleaning process. The aim of this report is to provide an evidence based review of the current and international practice standards guiding the selection, use and decontamination processes of UV facial shields in phototherapy. The complications and concerns that the COVID‐19 pandemic has had on this practice is highlighted. As such, we performed a comprehensive evaluation of the literature to provide recommendations as to the most effective, time efficient and safest practices for disinfection and decontamination of UV facial shields used in phototherapy during these unprecedented times. The range of face shields and visors available is vast, but despite this no guidelines have been published outlining the recommended specifications to consider in this process. In the era of COVID‐19, the literature has become populated with extensive and novel disinfection and decontamination practices for PPE. However, in phototherapy care consideration into this practice is required not only to ensure the eradication of the virulent virus to prevent spread but to ensure that UV protection standards of the PPE used, is not damaged during this process.What does this study add?No standardised guidelines or protocols regarding the selection process or decontamination process of PPE or face shields used in phototherapy published to date. During the COVID‐19 pandemic, PPE preservation and decontamination processes underwent significant scrutiny. In phototherapy the PPE requires extreme care, as incorrect disinfection practices can result in structure damage to the equipment reducing UV protection to the user. We report a comprehensive review and guideline into the specifications and selection process involved in PPE and face shield selection in phototherapy in addition to outlining the most safe and effective methods of UV‐PPE decontamination, in the era of COVID‐19.2UV‐radiation (UVR), commonly used for many dermatological conditions can lead to potentially damaging and harmful biological effects. As such, it is recommended that UV protective facial shields in combination with other forms of PPE be worn to protect both skin and eyes during treatment. There is a paucity in the literature guiding the selection process and safety specifications required for the PPE and facial shields worn in phototherapy. With a vast number of facial shields available, it is incumbent that all phototherapy departments are aware of the safety qualities and standards required to provide optimum protection for patients undergoing treatment to avoid unnecessary harm.To date, there are no standardised protocols or guidelines regarding the decontamination methods used for UV facial shields or PPE in phototherapy. With the rapidly emergent nature of the novel Coronavirus Disease 2019 (COVID‐19) there has been a concerted effort to find viable means of conserving PPE, including disinfection after use. As the incidence of COVID‐19 exponentially increases so too has the demand for PPE causing concern with supply meeting demand. To manage this, the literature became populated with novel innovative methods of preserving PPE, with new disinfection and decontamination processes. However, the UV‐PPE used in phototherapy requires extra care and caution as incorrect disinfection practices can result in damage to the structural integrity of the equipment impacting its protective qualities.Herein, with this report we provide guidance for phototherapy units on the safety standards and specifications required of UV facial shields employed in phototherapy. Following a comprehensive evaluation of the literature, we divulge the optimum disinfection and decontamination practices used for UV facial shields with reference to the challenges generated by the COVID‐19 pandemic.3An initial survey was undertaken of the practices and policies of our local phototherapy units, to determine the selection process and disinfection practices employed in terms of UV PPE, with reference to the COVID‐19 pandemic.A comprehensive literature review was undertaken, to identify international standards, guidelines, and policies to compare with local practices. References identified though the Medline, PubMed, Embase and CENTRAL databases. Scientific studies and research reports in peer‐reviewed journals were identified using relevant medical subject headings (MeSH) and field codes defining our clinical question. In addition, data from policy studies and briefs published by organisations including the Central Disease Control (CDC), World Health Organisation (WHO) were complied.The eligibility criteria used for our literature review process involved the ‘population, interventions, comparisons, outcomes and study type’ guideline. Our population cohort included all adults (over 16 years) undergoing UV phototherapy for dermatological skin conditions requiring the use of UV‐PPE face shields during the COVID‐19 pandemic period, dating January 2020 to September 2020. The intervention involved all decontamination practices, including chemical and physical disinfection processes. Comparisons were possible evaluating different disinfection methods and practices used in different healthcare facilities and research units. The outcome was disinfection efficacy, viral (COVID‐19) neutralisation and the maintenance of the UV‐PPE structural safety for the users. There were no restrictions placed on study type, owing to the limited data available on this subject matter.44.1A thorough evaluation of the current UV facial shield products available was performed, with reference to the various facial shield specifications and recommendations to consider in a selection process. The literature explores a number of factors to ruminate, including the manufacturing visor material, the comfort and fit, the UV waveband protection in addition to the anti‐fog, anti‐glare and reflection qualities and the compliance with international certification standards.4.1.1Typically, facial shield visors are composed of durable robust material like polycarbonate (PC), acetate, polyethylene‐terephthalate glycol (PETG) or less commonly steel or nylon mesh. Additionally, the visor is commonly treated with advanced coatings to offer anti‐glare, anti‐fog and anti‐scratch properties depending on the intended use or application of the facial shield.4.1.2The structural components of the facial shield involve the visor material as described, the frame mount and suspension system. Most phototherapy units advocate for a lightweight plastic frame with an adjustable mount for individual fit, with some manufacturers offering a detachable mechanism for ease of cleaning. It is recommended from the Centre of Disease Control and Preventions (CDC) that the dimensions of all UV facial shields should span beyond the full length of the patients face including chin and crown areas and extend to the lateral aspect of the ears at a minimum.4.1.3UVR as part of the electromagnetic radiation spectrum, displays wavelengths ranging from 100 to 400 nm, shorter than visible light and thus carrying more energy. At specific wavelengths, UVR can exert biological effects at molecular levels which can lead to observable clinical effects. UVR wavelengths are broadly subdivided into three specific bands: Ultraviolet‐A (UVA) ranging from 315 to 400 nm; Ultraviolet‐B (UVB) from 280 to 315 nm; and Ultraviolet‐C (UVC) 100–280 nm.4.1.4There are a variety of international certification awarded to UV PPE that meet and comply with specific standards. These qualifications have been published extensively by European committee for standardisation (CEN) awarding the European norm (EN) and the European economic area certification (CE) and by the American national standard institute (ANSI).4.2In the current COVID‐19 pandemic, concerns regarding the durability, reusability and decontamination practices of the UV facial shields in phototherapy became rising concern. Questioning the efficacy of current decontamination practices and the ability of the facial shields to withstand repeated cleaning processes, to ensure the eradication of the virulent virus whilst maintaining integrity of the shields structure.4.2.1Disinfection and decontamination processes for UV facial shields are guided by the manufacturing company. This process can involve the use of a plain soap and water, an environmental protection agency (EPA) approved or similar disinfectant or cleanser or an isopropyl based alcohol disinfection agent.Few investigative studies have reported favourable results regarding the use of soap and warm water as a decontamination process for PPE and UV facial shields. These studies outline methods of submerging the facial shield in warm soap water and using a gentle soft micro‐fibre cloth to wipe the surface clear. However, comparative analysis between these studies is confounded by the global differences of the variety of ingredients used in the soaps as well as the inconsistent techniques used in terms of water temperature and duration of submersion in addition to the diverse range of training facilities available to those performing the decontamination process.Surface disinfectant solution, spray or wipes are often used for UV facial shield cleaning process. Cleansing products that contain ammonium or that are high alcohol content must be used with caution as may lead to irreversible damage to the structure of the facial shield compromising the UVR protection. Similarly, there is a reported risk with these products creating visible film or residue on the surface of the facial shield that if not rinsed or cleaned in a timely fashion, can impair visual quality.There are very few studies analysing the effectivity of these techniques; soap, water or disinfection solution on UV facial shields to eradicate COVID‐19In early March 2020, the environmental protection agency (EPA) released the initial ‘List N’ of disinfectants for use against SARS‐CoV‐2. The qualifying germicidal, disposable Sani‐cloth wipes revealed a 3‐log reduction against COVID‐19 following a 1 min contact time. As such, offers a safe, effective and time‐efficient method of disinfection for UV facial shields. However, to avoid any surface scratches it is recommended to pre‐rinse or submerge the facial shield in water to dislodge any particulate matter or debris.4.2.2Vaporised hydrogen peroxide (VHP) is an environmentally safe and commonly used industrial and healthcare decontamination process used in the sterilisation of reusable metal and non‐metal devices as well as medical instruments and materials. The ease of generation of VHP at low controlled temperatures in addition to its rapid cleaning cycle (30–45 min) makes it a potential method of efficient facial shield decontamination. When compared with other sterilisation or decontamination practices including UV‐Germicidal Irradiation (UVGI), VHP is reported to disinfect all crevices and cornered areas of the facial shield exteriors, thus a more thorough and effective result.Amidst the ongoing pandemic, more recent studies have evaluated the effectivity of VHP in the disinfection of masks or facial shields contaminated with SARS‐CoV‐2. Kumar et al. report complete eradication of SARS‐CoV02 virus on N95 FFR following a 1 h treatment process involving 10 min if dehumidification, 3 min conditioning, 30 min of decontamination and 20 min of aeration. The peak VHP concentration recorded was 750ppm, with the FFR withstanding 10 cycles of this treatment without any compromise to mask performance or structure. Similarly, Smith et al report no functional degradation to the FFR following two cycles of VHP treatment and no recoverable or viable SARS‐CoV‐2 virus.4.2.3−2 of UVR is reported as the highest recommended dose for which no physical or structural damage to the facial shield is observed.−2, required to eliminate approximately 90% of single‐stranded‐RNA viruses, like COVID‐19 on gel media.−2 for 20 min on each side. Whilst a promising and convenient disinfection method is outlined, the dose of UVR at different levels within biosafety cabinet varied considerably thus stringent quality control and monitoring measures are required to ensure the safety of the shields is preserved.UVGI is another disinfection process that has been extensively explored throughout the COVID‐19 pandemic. UVGI uses wavelengths between 180 and 320 nm to disrupt DNA and RNA cross‐linking, preventing pathogenic replication. At this wavelength, the UVR dose is directly proportional to the inactivation and elimination of the surface pathogens.5In the absence of a clear consensus on the specification requirements of facial shields we provide a comprehensive review into the features and qualities that should be taken into consideration for all facial shields that are to be used in a phototherapy unit.The COVID‐19 pandemic has led to development of several novel, innovative disinfection processes for PPE, including the use of VHP and UVGI. Whilst remain promising techniques they are not, at present easily or readily available in most healthcare or phototherapy units. In addition, the financial weight associated with these systems, in terms of staffing and equipment resources, requiring continuous specialised operational training and maintenance is high. We recommend from this review, the use of simple but thorough disinfection with soap, water and soft micro‐fibre cloths or germicidal disposable ‘Sani‐Cloth’ wipes. Regular staff training, educational appraisals and techniques reviews guided by infection control, microbiology and medical physics departments, are required to ensure safety standards are maintained.With respect to ongoing planning for COVID‐19 we propose a process whereby each patient will receive their own designated facial shields for the duration of their treatment process. Between therapy sessions, simple disinfection measures will be taken as outlined and storage will be on site in patient specific lockers and safety cabinets. Following the completion of the phototherapy course, each facial shield should then undergo a comprehensive decontamination process prior to use by the next individual.Certainly, going forward the use of VHP or UVGI should be a consideration in all healthcare facilities as well as phototherapy departments as a potential long‐term comprehensive decontamination solution to the COVID‐19 pandemic. However, further research is required into the safety, efficacy and costing as well as the overall procedural approach and technique of these methods before formal introduction into clinical practice.The authors declare they have no conflicts of interest.Aoife Granahan: Conceptualization‐Equal; Data curation‐Lead; Investigation‐Lead; Methodology‐Lead; Project administration‐Lead; Writing – original draft‐Lead; Writing – review & editing‐Lead. Jackie Mc Cavana: Data curation‐Supporting; Formal analysis‐Supporting; Investigation‐Supporting; Supervision‐Equal; Validation‐Supporting; Writing – review & editing‐Supporting. Aoife Lally: Conceptualization‐Supporting; Investigation‐Supporting; Methodology‐Supporting; Supervision‐Equal; Validation‐Supporting; Writing – review & editing‐Equal. Imelda Morgan: Data curation‐Supporting; Formal analysis‐Supporting; Investigation‐Supporting; Supervision‐Supporting; Writing – review & editing‐Supporting. Susan Fitzgerald: Data curation‐Supporting; Investigation‐Supporting; Supervision‐Supporting; Writing – review & editing‐Supporting. Blaithin Moriarty: Conceptualization‐Equal; Data curation‐Supporting; Formal analysis‐Supporting; Investigation‐Supporting; Methodology‐Supporting; Supervision‐Lead; Validation‐Equal; Writing – original draft‐Supporting; Writing – review & editing‐Lead.


We report a case of intestinal lesions in a patient with a history of lupus nephritis and renal transplantation. Biopsy revealed an EBV‐driven post‐transplant lymphoproliferative disease (PTLD). An EBV‐driven PTLD is a major complication after renal transplantation and is an important differential diagnostic consideration in the follow‐up of renal transplant recipients. Post‐transplantation lymphoproliferative disease (PTLD) is an important diagnostic consideration in a transplant recipient that presents with intestinal disc‐like lesions. Her medical history was remarkable for lupus nephritis for which she had undergone a renal transplantation that was complicated by a severe rejection and treated with methylprednisolone, plasmapheresis, intravenous immunoglobulin, and antithymocyte globulin (ATG). Colonoscopy showed several disc‐like lesions with central ulceration in the colon as depicted in Figure 22.1Light microscopy of the biopsy specimens demonstrated infiltration of large lymphoid cells Figure  that werPost‐transplantation lymphoproliferative disease is a serious complication of renal transplantation.Post‐transplantation lymphoproliferative disease is an important differential consideration in a transplant recipient that presents with intestinal disc‐like lesions.Established risk factor for PTLD is cumulative amount of immunosuppression.Post‐transplantation lymphoproliferative disease is a serious complication of organ transplantation with a spectrum ranging from clinical indolent to aggressive lymphomas.Not applicable.Arjan J. Kwakernaak has co‐written the manuscript. Rob W. van der Pluijm has co‐written the manuscript. Neelke C.van der Weerd was medical supervisor during admission of the patient, provided important intellectual content and reviewed the manuscript. Lianne Koens provided details regarding the pathology findings, provided important intellectual content, and reviewed the manuscript. Wytske M. Westra and Krisztina B. Gecse provided details regarding the endoscopy findings and reviewed the manuscript. Ankie Kleinjan provided important intellectual content and reviewed the manuscript.We adhered to the ethics guidelines. Written informed consent was obtained from the patient to publish this report in accordance with the journal's patient consent policy.Written informed consent was obtained from the patient to publish this report in accordance with the journal's patient consent policy.


Community‐based cohort studies have shown that low levels of N‐terminal pro‐B‐type natriuretic peptide (NT‐proBNP) increase the risk of cardiovascular events and death.et al.In a recent issue of the mother journal, Verbrugge special patient population, and it is unclear whether the findings can be applied to patient populations that were excluded, even with the same disease phenotype. The treatment effects of some drugs that have not been shown to be effective in these special HFpEF populations are also uncertain in non‐special HFpEF populations.Recently, clinical trials of newer HF drugs, such as sacubitril valsartan and sodium‐glucose cotransporter 2 inhibitors, have pried open the tightly closed doors to effective treatments for the HFpEF population.special patient populations are often used to develop treatment recommendations in relevant guidelines and consensus statements. We currently have little evidence to manage the clinically important but overlooked HFpEF population, including patients with lower NP concentrations.Importantly, we need to be reminded of the current framework in which the findings of clinical trials conducted on et al.,cannot‐miss HFpEF phenotype.In the study by Verbrugge


To provide the first profile of the demographic and service characteristics of young people (aged 12–24 years) who access Foundry, a provincial network of integrated youth health and social service centres in British Columbia, Canada and to share early learnings about implementation and service innovation.Using a retrospective chart review, we conducted a census of all young people accessing a Foundry centre in a ‘proof of concept’ phase. Six centres were assessed between October 2015 and March 2018. Data included demographics, mental health service access history, service type the youth was seeking, and information about how they found out about the centre.A total of 4783 young people presented during this proof of concept period, for a total number of 35 791 visits. The most frequently accessed category of service was mental health/substance use (57%) followed by physical health (25%). Young people were most likely to be female, aged 15–19, and White. Youth demographic characteristics showed an over‐representation of Indigenous and LGBTQ2 youth and under‐representation of males and youth aged 20–24. Youth were most likely to learn about Foundry from a friend (44%) or family member (22%). Most youth (58%) reported that they would have gone ‘nowhere’ if not for Foundry.Foundry is a model of integrated health and social services delivery, focused on early intervention, prevention and accessibility, driven by the needs and priorities of young people and their families. Leveraging international integrated youth health service evidence, the model addresses urgent priorities in Canadian health service delivery. BC has a population of 5.5 million, with 2.3 million living in Greater Vancouver, the third‐largest metropolitan area and one of the most ethnically and linguistically diverse cities in Canada  to share learnings from their organizations to support the development of Foundry's proof of concept and evaluation framework  was established to oversee the complex process of establishing Foundry centres, beginning with the selection of communities via a formal request for proposals. Twenty‐five organizations representing communities from across BC responded. Working with the FCO, five Proof of Concept centres  were opened in each of the BC health authorities between (1 October 2015–31 March 31 2018). A developmental evaluation of the Foundry implementation process, including the work done by the Foundry Central Office, captured how Foundry impacted the young people and families who accessed services . It is also anticipated that the results from this evaluation will support jurisdictions creating similar models, including provincial leaders in Canada. Internationally, we hope this knowledge contributes to a growing body of new evidence about integrated youth services.22.1Data were collected on all young people accessing services at six operational Foundry centres, during the period of October 2015 to March 2018. All centres reported service count and type data. In four of six centres (excluding Vancouver and Abbotsford), demographic data were collected from each youth at the time of their first visit to a Foundry centre and in each subsequent visit (if they returned). Data were entered into a secure data management platform and aggregated for analysis. Under the Tri‐Council Policy Statement on Ethical Conduct for Research Involving Humans, this evaluation data was distinguished from research requiring research ethics board review as they were collected as part of a quality assurance study of a proof of concept in a ‘real world’ setting.2.2The forms captured basic demographic data , information about the type of services sought , information about how the youth found out about Foundry and where they would have sought services had a Foundry centre not been available to them.2.3n/N, %) for categorical variables.We analysed data using R . The age and gender distribution of youth accessing services showed a similar pattern for male and female clients aged 12–14 years and 20–24 years. A relatively higher proportion of female youth accessed services in the 15–19 age range (from 3% to 5% more); however, this difference was accounted for by higher utilization of sexual health services by females in this age range.As shown in Table 3.1.2Two‐thirds (66%) of young people self‐identified as White, followed by young people that identified as indigenous 14%). As shown in Table 4%. As sh3.1.3One in three (30%) youth accessing Foundry services self‐identified as having a sexual orientation other than heterosexual. Data from the Canadian Community Health Survey  reported living in insecure housing situations and 13% reported not having any income source. The vast majority of youth (91%) reported parents or other family members as their guardian. Notably, 7% reported that their guardian was a social worker, service agency or other. Only 1% of youth reported living with foster parents. Youth aged 18–21 were more likely to report having insecure housing. Youth who identified as male, transgender or other were more likely to report having insecure housing than females.3.2As shown in Figure 3.3Given the early focus on co‐design, Foundry sought to measure referral patterns, hinting at acceptability as well as an understanding of what youth viewed as help‐seeking alternatives to Foundry. Data showed that the most common referral sources for young people were there peers and their family members (45%), that is, lay people, signalling a significant and early credibility with the target population. Other referral sources were from family physicians (14%), school counsellors (13%), and the internet (12%).3.4As a way of exploring unmet need and early intervention, the question ‘where would you have gone today if not Foundry’ showed that nearly half (44%) responded ‘nowhere/I wouldn't have gotten help’. Another 22% reported that they would have turned to their family or friends  for assistance. Additionally, the majority of youth (58%) who accessed Foundry during the proof of concept phase reported that they had not accessed a mental health or substance use service in the past year. This outcome is highly suggestive of acceptability and addressing unmet need early.4Within the proof of concept period, Foundry centres have been successful in reaching the target age group of youth between the ages of 12 and 24 primarily seeking support for MHSU use concerns. It is encouraging that the highest levels of service utilization are from youth ages 15–19, the age of onset for many mental health issues . This variation highlighted the importance of having an integrated one‐stop health and social service delivery tailored to address the varying needs of young people across the 12–24 age range  (Government of British Columbia, Our shared learnings also include an understanding that future youth and family engagement requires organizations to create opportunities for input on strategies to draw in those young people choosing not to enter Foundry centres. Although the collection of metrics such as service access, service type, and demographics is important, an aspirational outcome of the Foundry initiative is to show transformation to access, notably by creating increased availability in part through service co‐location and integration and the development of low barrier services such as peer support and solution‐focused brief therapy, well suited for walk‐in counselling. Improved access or system integration were not measured here and future work with young people and communities is needed to co‐develop ways to capture effective integration and community engagement efforts.Finally, our findings contribute to an emerging body of literature on integrated youth health services (Hetrick et al., 4.1Our team learned that collecting data across a network of centres with individual electronic medical records within unique health authorities posed challenges. A customized electronic data collection platform, with strong data quality assurance and data governance measures, is needed to support consistent and meaningful data collection, providing a common registry for all youth and caregivers interacting with Foundry centres and allowing for the collection of a common evaluation and quality improvement data set and network practice guidelines that can be rolled out across the network of services. Other limitations include the mixed methodology used to collect the data. Given the absence of a common data collection tool, data were collected from several sources including paper forms and electronic medical record systems. Additionally, data fields for service utilization were not uniform for the centres. For instance, at one centre, substance use visits were coded as mental health visits, leading to under‐representation of one and over‐representation of another. Access data is also reflective of the supply of a given service rather than the demand. For instance, initial funding constraints have limited primary care availability, which is reflected in the service utilization data. As Foundry's service model matures, ongoing systematic data collection, including regular measures of youth outcomes and experience of care, will support the centres to improve service provisions in areas where youth report lower levels of satisfaction. Routine use of youth feedback will also ensure that Foundry maximizes the engagement of young people in having a say in their own care, a key component to Foundry's overall approach to care and consistent with its guiding principles. To avoid a strong selection bias of youth already using the centre, sampling young people who opted not to return or never to access a centre, would be more helpful to inform access.5This study provides early evidence that Foundry is establishing itself as a branded network of youth‐friendly, inviting, accessible centres that are bringing together numerous partners to deliver a wide range of services to meet the needs of young people and their caregivers. The investment by government, philanthropists and stakeholders has led to a reduction in accessibility barriers to an array of services supporting the health and wellness of young people and their families.The authors declare no conflicts of interest.


Dasyurus hallucatus), an Endangered marsupial carnivore. We used next generation sequencing methods to genotype 10,191 single nucleotide polymorphisms (SNPs) in 352 individuals from across a 3220‐km length of the continent, investigating patterns of population genomic structure and diversity, and identifying loci showing signals of putative selection. We found strong heterogeneity in the distribution of genomic diversity across the continent, characterized by (i) biogeographical barriers driving hierarchical population structure through long‐term isolation, and (ii) severe reductions in diversity resulting from population declines, exacerbated by the spread of introduced toxic cane toads (Rhinella marina). These results warn of a large ongoing loss of genomic diversity and associated adaptive capacity as mammals decline across northern Australia. Encouragingly, populations of the northern quoll established on toad‐free islands by translocations appear to have maintained most of the initial genomic diversity after 16 years. By mapping patterns of genomic diversity within and among populations, and investigating these patterns in the context of population declines, we can provide conservation managers with data critical to informed decision‐making. This includes the identification of populations that are candidates for genetic management, the importance of remnant island and insurance/translocated populations for the conservation of genetic diversity, and the characterization of putative evolutionarily significant units.Mammal declines across northern Australia are one of the major biodiversity loss events occurring globally. There has been no regional assessment of the implications of these species declines for genomic diversity. To address this, we conducted a species‐wide assessment of genomic diversity in the northern quoll ( The northern quoll is a small (<1500 g) generalist marsupial predator  in Australia , and populations are being actively managed by various jurisdictions. Because the spread of cane toads has so far proved impossible to halt, current management actions are aimed at mitigating the impact of toads. For example, the Northern Territory government established insurance populations on two offshore islands in 2003  as well as loci that are significantly associated with the presence of cane toads. We predict that our genomic analyses will identify (i) population genomic structure resulting from vicariance across biogeographical barriers; (ii) patterns of drift associated with isolated island populations and (iii) large impacts of cane toads, or other drivers of population decline, on population genomic diversity, with population bottlenecks in toad‐exposed populations potentially contributing to a reduction in genetic diversity. Our temporal replication will also enable us to directly observe the effect of establishing insurance populations on offshore islands in the short term (16 years). We suggest that this study will provide a quantitative example of the impacts that invasive species can have on the patterns of genomic diversity in native species, the conclusions of which will benefit attempts to mitigate the impacts of invasive species.22.1We collated tissue samples from across the entire species distribution from multiple personnel who had been involved in mammal trapping between 1993 and 2019, covering the regional jurisdictions of Queensland (QLD), the Northern Territory (NT) and Western Australia (WA) Figure . Four ofFor each sampling locality, we determined whether cane toads were present at the date of collection by cross‐referencing with published timelines of cane toad spread  were prepared and extracted in plate format (Qiagen DNeasy 96 Blood & Tissue Kit) following the standard protocol with an extended lysis (incubation at 56°C for 2 h then reduced to 37°C overnight). Samples consisting of dried blood spots on filter paper were extracted with a slight modification to the lysis step  digestion using PstI and NlaIII, (ii) ligation with one of 48 unique inline barcoded adapters compatible with the restriction site overhang, (iii) manual sample pooling, (iv) DNA purification (QIAquick PCR Purification Kit followed by SPRIselect paramagnetic beads), (v) size‐selection targeting fragments of 280–375 bp in size  and (vi) a PCR (polymerase chain reaction) amplification step where one of two multiplexing index primers was added. Indexed libraries were pooled together and loaded onto flow cells for 150‐bp single‐end sequencing on either an Illumina NextSeq 500 (three plates) or a NovaSeq 6000 (two plates) platform.Double‐stranded DNA concentrations were quantified using a Qubit 3.0 Fluorometer and normalized to 200 ng DNA in 25 μl, and then arranged on 96‐well plates, where each plate contained two within‐plate technical replicates, one technical replicate from each of the other plates and one negative control (blank) well. Plates were sent for double‐digest restriction‐associated DNA (ddRAD) sequencing at the Australian Genome Research Facility in Melbourne, Victoria. An optimal combination of two restriction enzymes was determined using three establishment samples (broadly representative of the species distribution), with 2.3process_radtags function of stacks  >0.6 , and then pruned SNPs based on linkage disequilibrium via the “SNPRelate” package  the level of missing data; (ii) observed heterozygosity; (iii) minor allele counts and (iv) whether they showed signals of being under selection. We filtered individuals based on (i) the level of missing data and (ii) relatedness to other genotyped individuals within sampled localities. To filter SNPs and samples based on the level of missing data, we used a custom script to iteratively remove individuals and SNPs with decreasing missing data thresholds until no SNP had more than 3% missing data and no individual had more than 20% missing data. We then removed any SNP with a minor allele count <3 . One of each pair was then removed from all further analyses. The data set was then further pruned to remove highly related individuals and SNPs showing signals of putative selection, both of which can bias analyses that were developed using the assumption of unrelated individuals and neutrally evolving loci.To ensure that relationships between individuals could be accurately inferred from the data, we used these SNPs and samples to construct a hierarchical clustering dendrogram based on genetic distance, with visual examination of the dendrogram confirming that all technical replicates paired closely together on long branches Figure . We alsoTo estimate relatedness between individuals, we used the recently developed method‐of‐moments estimate for the kinship coefficient , and identification of SNPs that were significantly associated with the presence of cane toads (via a latent factor mixed model). To identify outlier SNPs that showed signatures of diversifying selection, we first computed an ancestry matrix for each individual using the methods implemented in the LEA package  from the data set that were identified using either method. This produced a selectively neutral data set for population genomic analyses, many of which rely on assumptions of neutrality. While future work may attempt to further elucidate and understand the patterns and genomic architecture of selection in response to cane toad invasion, the set of candidate loci identified here  providesAfter all filtering steps had been conducted, we retained a total of 10,191 SNPs and 352 individuals for the remaining analyses. The overall level of missing data for the final filtered SNP‐by‐individual matrix was 1.2%.2.5A), effective number of alleles (AE), observed heterozygosity (HO), expected heterozygosity (HE), Wright's inbreeding coefficient (FIS) and the locus polymorphic index (PE). The East Alligator site was separated into toad‐naïve (2003) and toad‐exposed (2014 and 2017) subsets prior to calculations being made. Similarly, the historical (2005) samples from Astell Island were separated from the more recent (2016) samples. Calculations were made using the “gstudio” package , and analysing the data set using the core program “Populations” with all missing sites removed. The heterozygosity estimates and standard errors in the subsection of the summary output titled “# All positions (variant and fixed)” were recorded, and Pearson's product moment correlation coefficient was used to determine the strength of the relationship between values of autosomal heterozygosity and SNP heterozygosity.Using the filtered SNP data set, we calculated mean values of standard genomic parameters for each sampling locality within each treatment, including the number of alleles  of ancestral populations, allowing for independent investigation of values for k that have low cross‐entropy metrics  across the landscape. To prevent interpolation of genomic structure beyond the geographical range of the northern quoll, we limited the interpolation surface to the known historical (pre‐European) distribution of the species.To assign individuals to ancestral population genomic clusters, investigate patterns of admixture between populations and assess hierarchical population structure, we used the alternating projected least squares algorithm implemented in the “tess3r” package  and total gene diversity (HT) to improve inference of demographic history among populations . A Mantel test was used to test the significance of geographical distance on standardized genomic differentiation, using 10,000 permutations. Isolation‐by‐distance among all mainland individuals was also investigated using spatial autocorrelation of multilocus genotypes, plotted on a correlogram. Pairwise geographical distances between individuals were calculated from latitude and longitude observations for each individual via the earth.dist function of the “fossil” package  or toad invasion and establishment  had on genomic diversity metrics, we constructed individual generalized linear models for each metric, where loci were samples and the predictor variable was a factor corresponding to the year of the survey.To test the relative influence of vicariance and cane toads on genomic diversity among our natural treatments , we constructed four generalized least square models using the “nlme” package  using three methods, of which one is appropriate for estimating recent  trends and two that are most useful for deeper time  trends.To identify patterns of historical demography at a locality, including the potential influence of cane toads on population size, we investigated past changes in effective population size (Ne), we used the version 1.1 software tool  file output from angsd was split into individual VCF files for each sampling locality, with plink version 1.9  were plotted and the x‐axis scaled to time assuming a mean generation time of 2 years  that estimates demographic histories based on the composite likelihood of a population's folded site frequency spectrum , we constructed a folded site frequency spectrum using genotype likelihoods estimated by the angsd software package. All filtering criteria based on mapping and read qualities were the same as previously described, with the filters for polymorphic loci removed to capture information on all monomorphic and polymorphic nucleic sites. By using likelihoods, the site frequency spectrum could be built from between 121 million and 565 million nucleic sites per locality. The site frequency spectrum was loaded into the folded blueprint file provided with the software, with an appropriate mutation rate per site per generation , whereas others were considerably more diverse    . Autosomameters) .z = 38,952, p < .001). Analysis of spatial autocorrelation of genotypes identified significant values of the autocorrelation coefficient r persisting for distances exceeding 500 km (FST = 0.5–0.9), probably due to its very low level of diversity (FST had a significant (p < .001) level of differentiation, based on 1000 bootstraps across loci, even where the same locality was sampled only a few years apart . The FST values were very similar to the GST″ values .Patterns of pairwise genomic differentiation across the landscape broadly corresponded to the pattern of geographical distance between localities , with thg 500 km . Seawateiversity . All pai″ values , with th3.3HE , AE  and PE  all differed significantly among the three  treatments. There was no significant difference in FIS among treatments . Tukey post hoc tests showed that toad‐naïve localities (mean HE = 0.049) had significantly higher HE than toad‐exposed (mean HE = 0.024) and island (mean HE = 0.019) localities , with no significant difference between island and toad‐exposed localities . Values of HO, AE and PE among treatments showed similar patterns to HE (AE = 1.072) had significantly higher AE than toad‐exposed (mean AE = 1.035) and island (mean AE = 1.03) localities , with no significant difference in AE between island and toad‐exposed localities .We found that three of the analysed population genomic parameters varied significantly among treatments Figure . The gens to HE . For exaFIS value for the SNP data set increased from 0.01 to 0.04 . This is despite only minor changes in HE  and HO  over the same period. At the East Alligator locality between 2003 and 2017, where cane toads invaded between sampling periods, there was a significant reduction in PE  and HE , but no change in HO  or FIS , based on the SNP data. Heterozygosity estimates calculated using the SNP data broadly agreed with heterozygosity estimates calculated from autosomal data . However, the estimates calculated from SNP data showed slightly greater stochasticity than the autosomal estimates, probably due to sample size variation among populations/years. For example, SNP estimates of HE at East Alligator increased slightly from 2003 to 2014, and then decreased from 2014 to 2017, whereas the autosomal estimates show a consistent reduction through time in HE and HO, with both values declining from 7e−5 to 6e−5 between 2003 and 2014, and no change observed between 2014 and 2017. As the autosomal estimates are likely to be a more robust measure of heterozygosity, we suggest this pattern is more realistic.Temporally replicated localities showed some significant changes through time, corresponding with isolation on islands or toad invasion and establishment Table . For exasnep suggested that there have been strong declines in the effective population sizes (Ne) of many northern quoll populations for much of the last century  tended to show smaller but somewhat more consistent population sizes through time than mainland populations. The toad‐exposed QLD localities were characterized by small effective population sizes for much of the past century, with noticeable reductions in Ne after arrival of cane toads into Mareeba, Hope Vale and the combined Hope Vale/Black Mountain localities. However, there did not appear to be enough genomic diversity in the sequence data for the Weipa locality to provide snep with the information content required to see past the arrival of cane toads to the area.Investigation of historical demographics using y Figure . LocalitNe of island populations around the time of isolation due to sea‐level rise after the end of the last glacial maximum . The EBSPs showed considerably less certainty with estimated population sizes, highlighted by the large confidence intervals for all localities that were analysed and mediocre convergence (posterior effective sample size) in some localities . Results suggested recent population declines in most populations, consistent with the outputs from snep and Stairway Plot 2 . This was most pronounced for populations in the highly remote and rugged areas of the western Kimberley, where threats may have developed more recently.Stairway plots showed expected fluctuations in m Figure , suggesty Plot 2 , with thy Plot 2 . While w4While biodiversity declines are well recognized as a global problem, the consequences of these declines for patterns of genomic diversity are often poorly known. Using genomic data to infer recent and long‐term historical demographic trends can help identify the extent to which loss of genomic diversity itself is threatening species' persistence. Northern Australia's vast savannas are the location of some of the world's most dramatic recent declines of mammals   in toad‐exposed populations, it is worth noting that this metric does not provide information on the cumulative level of inbreeding in a population, but rather detects nonrandom mating in the most recent generation  could help quantify the relative importance of recent (<50 years) changes in landscape management as drivers of decline. For example, while feral cats have probably been present for ~150 years , Groote Eylandt (NT), eastern Kimberley (WA) and Cape York (QLD), respectively  that biogeographical barriers separate subregions each containing substantial amounts of unique genomic diversity; (ii) that remnant island populations harbour only a subset of the diversity present within each subregion; (iii) that large insurance populations on islands or in intensively managed/fenced safe havens can maintain high levels of genomic diversity  and (iv) that declines induced by cane toads are dramatic, but play out against broader declines resulting from other anthropogenic stressors and threats. With the exception of cane toad impacts, these considerations are likely to be similar for many of the widespread faunal species within the Australian monsoonal tropics . These challenges span agriculture, biomedicine, the environment and industry, as well as extending to relevant international endeavours.A research collaboration was developed with researchers around Australia, who provided tissue samples for genetic analysis. All collaborators were offered the opportunity to contribute to the manuscript as co‐authors, and the results of research are available for the broader scientific community through online databases and open access publishing. The research addresses a priority concern, in this case the conservation genetics and management of the threatened northern quoll, and findings will be incorporated into management actions for the species. Bioplatforms Australia is a nonprofit organization that supports Australian Life science research by investing in state‐of‐the‐art infrastructure and expertise in genomics, proteomics, metabolomics and bioinformatics. Investment funding for Bioplatforms Australia is provided by the Commonwealth Government National Collaborative Research Infrastructure Strategy. The Bioplatforms Australia Oz Mammal Genomics Initiative is one of many national collaborative projects that generate high‐impact data and knowledge resources to support some of Australia's biggest scientific challenges (Appendix S1Click here for additional data file.


Early‐life experience influences social and emotional behaviour in adulthood. Affiliative tactile stimuli in early life facilitate the development of social and emotional behaviour, whereas early‐life adverse stimuli have been shown to increase the risk of various diseases in later life. On the other hand, oxytocin has been shown to have organizational actions during early‐life stages. However, the detailed mechanisms of the effects of early‐life experience and oxytocin remain unclear. Here, we review the effects of affiliative tactile stimuli during the neonatal period and neonatal oxytocin treatment on the activity of the oxytocin–oxytocin receptor system and social or emotional behaviour in adulthood. Both affiliative tactile stimuli and early‐life adverse stimuli in the neonatal period acutely activate the oxytocin–oxytocin receptor system in the brain but modulate social behaviour and anxiety‐related behaviour apparently in an opposite direction in adulthood. Accumulating evidence suggests that affiliative tactile stimuli and exogenous application of oxytocin in early‐life stages induce higher activity of the oxytocin–oxytocin receptor system in adulthood, although the effects are dependent on experimental procedures, sex, dosages and brain regions examined. On the other hand, early‐life stressful stimuli appear to induce reduced activity of the oxytocin–oxytocin receptor system, possibly leading to adverse actions in adulthood. It is possible that activation of a specific oxytocin system can induce beneficial actions against early‐life maltreatments and thus could be used for the treatment of developmental psychiatric disorders. Article Summary: Early‐life experience modulates the activity of the oxytocin system and induces plastic changes in the oxytocin system. Activation of the oxytocin system in early life modulates behaviours in adulthood. Oxytocin exerts not only reproduction‐related functions such as delivery of children and milk ejection during the lactation period, but also various actions including control of stress‐related responses, emotional responses and social behaviour.Oxytocin acts within the brain to induce various acute adaptive responses to environmental challenges. Oxytocin exerts neurotrophic actions in some cases. Oxytocin also has organizational effects via acting at certain developmental stages to modulate the developmental trajectory. These oxytocin actions are exerted mainly by activation of the oxytocin receptor located on neurons, astrogliaVarious acute actions of oxytocin have been reported. Oxytocin facilitates the acquisition of social recognitionIn addition to acute actions, oxytocin has been shown to be involved in neurogenesis and neurotropic actions.Oxytocin has also been suggested to function during the developmental period.22.1The oxytocin–oxytocin receptor system appears to be established at a relatively late stage of development compared to the development of the vasopressin system.Because the oxytocin–oxytocin receptor system develops from the perinatal period until adulthood, experiences especially in early life may influence the development of the oxytocin system to modulate neuroendocrine, autonomic and behavioural responses to environmental challenges in adulthood. In the next section, the effects of tactile stimuli during early‐life stages on behaviours and the oxytocin system are discussed.2.2Tactile stimulation plays an important role in non‐verbal emotional communication.During the developmental period, tactile stimulation also plays an indispensable role. Nurturing touch during parental care has been proposed to facilitate the development of emotional and social behaviours and cognitive functionsWarm sensations in addition to mechanosensation form an important part of tactile stimulation during parental care. Maintenance of body temperature is important for survival during the developmental period when body size is small. Huddling behaviour effectively reduces heat loss. Rodents show thermal‐seeking huddling. It is interesting that oxytocin, which is increased in pups by contact with the mother or parental care,Not only affiliative gentle mechanosensory stimuliAs described above, the oxytocin system is acutely activated by affiliative parental care. Parental affiliative stimuli during early life have also been shown to induce long‐lasting influence on the oxytocin system. Early‐life manipulations such as handling, maternal separation, nursery environments and paternal deprivation influence the amount of parental care that pups receive. In the next section, the long‐lasting influence of parental care and its experimental manipulations of handling, maternal separation, cage environment changes and paternal deprivation on the oxytocin system is discussed.2.32.3.1Parental care affects expression of the oxytocin receptor in the brain in various animals.Affiliative stimuli received during post‐weaning period also induce long‐lasting stimulating actions on the oxytocin system. Repeated application of gentle stroking stimuli, which induce affiliative behaviour in both female and male rats toward the experimenter, provokes enhanced activity of oxytocin neurons in response to stroking stimuli.2.3.2Maternal deprivation has been shown in rhesus macaques to increase anxiety in response to isolation, and the sensitivity of maternal separation to induce augmented anxiety depends on a SNP of the oxytocin receptor gene,Several studies have shown that postnatal maternal separation reduces oxytocin mRNA, oxytocin content or the number of oxytocin neurons in the hypothalamus and decreases the expression of the oxytocin receptor in some brain regions, although differences depending on the species, sex, experimental procedures and brain regions examined have been reported Table .–1) has been shown to reduce the hypothalamic oxytocin content in suckling rats–1) has also been reported to induce a long‐lasting decrease in oxytocin mRNA or oxytocin content in the hypothalamus and/or in other brain regions in mice,Postnatal maternal separation for a short duration  or alloparental care in monogamous animals, have also been shown to affect social behaviours and the oxytocin system in adulthood. In monogamous mandarin voles and prairie voles, both fathers and mothers show parental care toward their children. Paternal absence has been shown to reduce partner preference formation of the offspring in adulthood,As summarized above, parental care has been shown to produce long‐lasting changes in the oxytocin system. Generally, oxytocin neuronal activity is activated after affiliative parental care and oxytocin receptor expression is increased in correlation with the amount of paternal care, although differential influences dependent on brain regions, experimental manipulation and sex have been reported.Oxytocin neurons have been shown to be activated not only by affiliative stimuli, but also by stressful stimuli in adulthood. During the developmental period, stressful stimuli as well as parental care acutely influence activity of oxytocin neurons. Stressful stimuli in early life have also been shown to induce long‐lasting changes in the oxytocin system,2.4In animals in adulthood, various stressful stimuli,During the developmental period, stressful stimuli influence oxytocin neurons in the hypothalamus. Social isolation stress during the lactating period has been reported to modulate oxytocin immunoreactivity in the hypothalamus of monogamous mandarin voles.The oxytocin system has been shown to be influenced for a long time after early‐life stressful stimuli including prenatal exposure to alcohol Table . PrenataPostnatal sensory deprivation has also been shown to decrease activity of oxytocin neurons in the hypothalamus. Sensory deprivation by whisker deprivation and dark rearing have been reported to decrease oxytocin mRNA in the hypothalamus and to reduce excitatory synaptic transmission in the sensory cortex in mice.Post‐weaning social isolation has also been shown to reduce activity of oxytocin neurons in the hypothalamus of female rats in response to exposure to novel conspecifics, to impair social preferenceAs discussed above, numerous animal experiments have shown that various stressful treatments during early‐life stages have long‐lasting inhibitory actions on the oxytocin system. In the next section, the effects of adverse environments in early life on the oxytocin system in humans are discussed.2.5In humans, prenatal or postnatal stressful stimuli have been shown to be associated with altered functions of brain regions including the superior frontal gyrus for cognitive processing, amygdala for emotional processing, precuneus for memory processing and putamen for reward‐related learning processing.Childhood maltreatment has been shown to increase DNA methylation and to modulate the expression of various genesEarly‐life experience has also been shown to modulate effects of oxytocin administration possibly by inducing changes in the oxytocin system in adulthood. The effects of oxytocin administration appear to be different depending on early‐life stress in humans. Intranasal oxytocin application reduced premenstrual emotional symptoms in a control group of females, whereas oxytocin administration augmented the emotional symptoms in women who had experienced early‐life abuse.As discussed in the above two sections, both animal and human studies have shown that stressful stimuli in early life acutely activate the oxytocin system and induce long‐lasting suppressive actions on the oxytocin system. In the next section, the role of oxytocin and effects of oxytocin administration in early life is discussed.2.6Oxytocin in early life has been shown to attenuate early‐life stress‐induced or genetically induced impairments of social behaviour or energy metabolisms, although detailed mechanisms of the organizational actions of oxytocin remain to be clarified.2.6.1The prenatal oxytocin system has been suggested to affect behaviour in adulthood. Male oxytocin‐deficient mice born to oxytocin‐knockout dams and fostered to wild‐type dams show augmented aggressive behaviour, whereas oxytocin knockout male mice born to heterozygous dams and fostered to wild‐type dams do not show heightened aggressive behaviour.2.6.2Perinatal oxytocin has also been suggested to affect behaviour in adulthood. At the time of delivery, a large amount of oxytocin is released in mothers and might act in both mothers and newborns.–1, postnatals day 10–14) has been reported to increasePlasma oxytocin concentrations are lower in caesarean‐section neonates and mothers than in vaginally born babies and mothers.Oxytocin has been widely used for stimulation of births. Prenatal oxytocin administration to mothers has been shown to affect various brain functions in adulthood. In prairie voles, pups who received exogenous oxytocin via administration to their mothers were reported to show increased plasma oxytocin and increased activation of oxytocin neurons, suggesting that maternally administered oxytocin crosses the placenta into the pup and activates hypothalamic oxytocin neurons of the pup. Adult male offspring of oxytocin‐administered dams have been reported to show increased expression of the oxytocin receptor in the central amygdala and insular cortex, to exhibit increased alloparental caregiving toward pups and to display close social contact with other adults.For stimulation of labour in humans, low doses of oxytocin are applied over a long period. In humans, a population‐based study showed that perinatal exogenous administration of oxytocin has no association with childhood emotional disorders,2.6.3Milk contains oxytocin.On the other hand, postnatal oxytocin application at large dosages has been reported to rescue social behaviour, pain‐related behaviour and memory in adulthood, which are impaired by genetic manipulations or behavioural treatments.–1 in the first 2 days or postnatal week) has been shown to increase pre‐ or post‐synaptic transcript levels of neurexins and neuroligins–1) during postnatal days 7–21 has also been shown to restore the number of hypothalamic oxytocin neurons and to reverse social deficits in contactin‐associated protein‐like 2 (CNTNAP2)‐deficient mice, another model of autism spectrum disorder.–1 in the first week) has also been reported to result in recovery of a decrease in oxytocin neurons and deficits in social behaviour in rats that had been prenatally treated with valpronic acid.Postnatal daily administration of oxytocin  in rats has been shown to induce hypersensitivity in response to nociceptive stimuli and to impair oxytocin receptor‐dependent stress‐induced analgesia and oxytocin receptor‐dependent anti‐hyperalgesia.–1 during postnatal days 1–14) reduces partner preference in adult female prairie voles, and female voles that show high oxytocin receptor binding in the nucleus accumbens have resilience to neonatal isolation‐induced impairments. These social isolation‐induced impairments in attachment behaviour are mitigated by neonatal treatment with melanotan‐II administration, which facilitates oxytocin release.Neonatal isolation  induces reduction in spatial memory and disturbance of long‐term potentiation in the hippocampus, and this reduction has been shown to be attenuated by repetitive application of oxytocin in rats.Chronic stress in early life caused by maternal separation  to switch GABA function from depolarization to hyperpolarization in an early and narrow developmental time window of life.3Early‐life experience changes the activity of oxytocin neurons and possibly oxytocin receptor‐expressing neurons at the time of experience, resulting in plastic changes in the expression of oxytocin and/or the oxytocin receptor in adulthood, although the magnitude and direction of changes appear to be dependent on early experience, genetic differences, brain regions and sex. Generally, affiliative experience activates the oxytocin system in early life and induces enhanced activity of oxytocin neurons and increased expression of the oxytocin receptor in adulthood. On the other hand, early adverse experience activates the oxytocin system in early life but induces reduced activity of oxytocin neurons and the oxytocin receptor in adulthood. The mechanisms underlying the opposite long‐lasting effects of affiliative and adverse experiences remain to be clarified. Oxytocin administration at moderate dosages in early life induces activation of the oxytocin system in adulthood, suggesting that the oxytocin system in early life has a positive feedback system to induce long‐lasting activation of the oxytocin system in adulthood. On the other hand, administration of a large amount of oxytocin in early life sometimes induces opposite actions in adulthood. The magnitude of activations of oxytocin neurons by affiliative stimuli is relatively small and the activation is transient, whereas stressful stimuli can drastically activate oxytocin neurons. Stressful stimuli stimulate not only the oxytocin system, but also other systems including the hypothalamic‐pituitary‐adrenocortical axis, sympatho‐adrenomedullary system, opioid system, allopregnanolone system and inflammatory responses. It is possible that overactivation of oxytocin neurons and/or other stress‐related systems activated during stressful stimuli in early‐life stages induces long‐lasting inhibitory actions on the oxytocin system in adulthood Figure .The effects of early‐life stimuli and oxytocin administration have been shown to be dependent on sex and brain regions examined. Female pups appear to be more sensitive than male pups to the neonatal manipulation of oxytocin,Inconsistent and sometimes contradictory data concerning the effects of early‐life experience and oxytocin administration have been reported. In an epigenetic landscape model, balls, which represent the brain, stand at a similar initiating point and roll downhill through bifurcating valleys, resulting in the balls reaching very different endpoints. Thus, small differences in experimental procedures for early‐life experience among different laboratories might have caused a large variation in the data for adulthood.It is also likely that discrepancies are caused, at least in part, by the complex nature of early‐life experience. For example, maternal separation has several components including sensory deprivation from mothers  and also exposure to novel environments during separation, possible nutritional deprivation and temperature loss. Maternal separation modulates the parental care of mothers afterwards. Some of these components may be variable depending on laboratories. Simpler stimulation of a mono‐sensory system and pathway‐specific activity manipulation using molecular techniques may be useful as an early‐life stimulus for clarifying the effects of early‐life experience.Oxytocin administered not only during the neonatal period, but also during the adolescent period has been shown to have long‐lasting prosocial, anti‐anxiety and anti‐aggression actions.Oxytocin has been considered for treatment of autism spectrum disorderThe authors declare that they have no conflicts of interest.Tatsushi Onaka: Conceptualization; Funding acquisition; Writing – original draft; Writing – review & editing. Yuki Takayanagi: Conceptualization; Funding acquisition; Writing – review & editing.


Single‐cell RNA‐sequencing (scRNA‐seq) is a powerful approach to unravel gene expression profiles of individual cells and has become an indispensable tool in understanding cells participating in physiological and pathological processes. This approach can discover new cell populations and genes functioning within a specific group of cells at a single‐cell level. Bones and other musculoskeletal tissues are no exception. An important consideration is that currently commercialized platforms require dissociated single cells in suspension to be loaded onto droplet‐generating microfluidic devices Fig .Given the technical challenge to isolate cells from the bone at a single‐cell level, the application of this powerful technology is still at an early phase in bone research field. One of the most tractable bone‐residing cell populations is the monocyte–macrophage lineage, which are of hematopoietic origin and precursors for bone‐resorbing osteoclasts.JBMR Plus, two original articles highlight the power of scRNA‐seq approaches in discovering new macrophage subtypes and identifying new genes functioning in osteoclast precursor cells of both humans and mice. In the first article by Fujii and colleagues, the authors identified a novel “surgery‐induced” highly inflammatory CD9+ IL1+ macrophage population in a mouse model of anterior cruciate ligament reconstruction surgery, which resolved early and became replaced by another type of macrophages. In the second article by Omata and colleagues, the authors identified a GTPase family member, RAB38, as a highly expressed molecule in both human and murine osteoclast precursor cells. This protein was associated with dynamic changes in histone modification and transcriptional regulation. Fujii and colleagues evaluated bone healing of mice lacking Ccr2, which they identified as one of the markers for late‐responding macrophages after the surgery. Omata and colleagues silenced Rab38 expression using short hairpin RNA and evaluated osteoclast differentiation in vitro. These initial validation studies placed the computational findings in a biological context, although much remains to be done to fully understand the implications of the large data sets.In this issue of lleagues,1 the JBMR Plus understands the need to publish exciting discoveries quickly. Rapid publication allows the entire research community to think about the data and use it in a variety of ways, advancing the field as a whole. Therefore, we welcome technically rigorous studies that describe large data sets, albeit with somewhat limited functional validation. The two articles in this issue describing scRNA‐seq data sets are great examples of the type of studies that JBMR Plus seeks to publish. Analogous studies employing analyses such as proteomics and metabolomics generating equally impactful data sets will also be welcomed. We hope that investigators in the musculoskeletal research community will utilize JBMR Plus to disseminate and discover important work in a timely manner.We appreciate that functional validation is a very challenging step, and comprehensive characterization of identified cells or proteins may take many months or even years. As a fully open‐access journal dedicated to rapid dissemination of important findings in musculoskeletal research,


To investigate the association between planned mode of birth after previous caesarean section and a child’s risk of having a record of special educational needs (SENs).Population‐based cohort study.Scotland.A cohort of 44 892 singleton children born at term in Scotland between 2002 and 2011 to women with one or more previous caesarean sections.Linkage of Scottish national health and education data sets.Any SENs and specific types of SEN recorded when a child was aged 4–11 years and attending a Scottish primary or special school.Children born following planned vaginal birth after previous caesarean (VBAC) compared with elective repeat caesarean section (ERCS) had a similar risk of having a record of any SENs  or specific types of SEN. There was also little evidence that planned VBAC with or without labour induction compared with ERCS was associated with a child’s risk of having a record of any SENs  or most types of SEN. However, an increased risk of sensory impairment was seen for planned VBAC with labour induction compared with ERCS .This study provides little evidence of an association between planned mode of birth after previous caesarean and SENs in childhood beyond a small absolute increased risk of sensory impairment seen for planned VBAC with labour induction. This finding may be the result of performing multiple comparisons or residual confounding. The findings provide valuable information to manage and counsel women with previous caesarean section concerning their future birth choices.There is little evidence planned mode of birth after previous caesarean section is associated with special educational needs in childhood. There is little evidence planned mode of birth after previous caesarean section is associated with special educational needs in childhood. Globally, caesarean section rates are thought to have almost doubled between 2000 and 2015.Caesarean section may adversely affect a child’s neurodevelopment through several mechanisms, including altering the composition of the infant’s gut microbiota,A population‐based cohort study was conducted using Scottish national health and education (pupil census) data sets  in Scotland, UK, between 1 January 2002 and 31 December 2011 to women with one or more previous caesarean sections were identified. Births to women not considered clinically eligible to plan VBAC based on current UK guidelines were excluded Table ,5, 6The main exposure was planned mode of birth after previous caesarean section, with planned VBAC  compared with ERCS . Additional analyses compared planned VBAC with or without labour induction and ERCS. Analyses were also performed by actual mode of birth after previous caesarean section, comparing children born by VBAC , those born by in‐labour non‐elective repeat caesarean section (birth by non‐elective caesarean with a duration of labour of ≥1 hour) and those born by ERCS.The primary outcome was any record of SENs and the secondary outcomes were the specific type of SEN, with children classified as having a record of SENs if it was recorded in any pupil census year when they were aged 4–11 years and attending a Scottish primary or special school (school providing education to children with complex or specific needs that cannot be met in mainstream schools). The Education  Scotland Act 2004 (as amended) defines SENs as being unable to benefit from the school education provided without the provision of additional support beyond that normally given to schoolchildren of the same age. Educational authorities have a legal duty to identify and provide support for children with SENs. For this study, SENs arising from the following causes were included: learning disability; dyslexia; other specific or moderate learning difficulty; sensory impairment; physical or motor impairment; language or speech disorder; ASD; social, emotional and behavioural difficulty or mental health problem; and physical health problem. A child can be recorded as having more than one type of SEN.All analyses were prespecified as described in the methods. Associations between the exposures and outcomes were examined using logistic regression to estimate odds ratios (ORs), where the outcome was rare (affecting <10% of population), and modified Poisson regression to estimate risk ratios (RRs),mp  version 16.Several sensitivity analyses were conducted. First, complete case analyses were performed. Second, as the criterion used to identify planned mode of birth could misclassify women who planned ERCS but went into labour before their scheduled caesarean date, analyses were repeated, limiting to the gestation recommended by UK guidelines to perform an ERCS (≥39 weeks of gestation).As part of a larger programme of research, this study was supported by patient and public involvement (PPI) representatives from various service user and voluntary groups in the perinatal field. PPI representatives provided input on the proposed research plan, including the research questions, when developing the funding application for the programme of research. PPI representatives have also commented on the findings so far and have contributed to the dissemination plan for the programme of research. This will include producing, with the help of the PPI representatives, a plain language summary leaflet of the research findings.In total, 50 025 live singleton term births to women with one or more previous caesarean sections met the eligibility criteria for the study. Of these, 5133 (10.3%) were excluded because they did not link to any pupil census records between 2007 and 2016, when the child was aged 4–11 years and attending a primary of special school  were born following planned VBAC and 26 041 (58.0%) were born by ERCS. The ERCS rate increased from 50.2% in 2002 to 64.7% in 2011. The mothers of children born following planned VBAC were more likely than those born by ERCS to be younger, born outside the UK and have a lower socioeconomic status  were born to mothers who had their labour induced . Outcomes following planned VBAC with or without labour induction, compared with ERCS, are presented in Table Of the children born following planned VBAC, 72.0% (13 564/18 851) were actually born by VBAC and 28.0% (5287/18 851) were born by in‐labour non‐elective repeat caesarean section. There was no strong evidence that actual mode of birth after previous caesarean section was associated with a child’s risk of having a record of any SENs or most types of SEN Figure . In the There was no evidence of effect modification by breastfeeding status at 6–8 weeks postpartum, the number of prior caesarean sections that the mother had, whether the mother had any prior vaginal birth, gestational age at birth or, in the repeated‐measures sensitivity analysis, the age of the child. The sensitivity analyses Tables  largely In this study of 44 892 children born to women considered clinically eligible to plan VBAC, planned VBAC compared with ERCS was associated with a similar risk of the child having a record of any SENs or a specific type of SEN when they were aged 4–11 years and attending a primary or special school. However, compared with ERCS, planned VBAC with labour induction was associated with a 60% increased risk of sensory impairment, although the absolute risk difference was small (0.4%). There was also evidence of a 24% reduced risk of language or speech disorder with in‐labour non‐elective repeat caesarean, compared with ERCS, although the absolute risk difference was again small (0.99%). As discussed subsequently, these differences may be the result of performing multiple comparisons or residual confounding.The study strengths include its large population‐based design, the use of national prospectively collected data subject to regular quality checks, restriction to children born to women considered clinically eligible to plan VBAC and the ability to explore the influence of many a priori covariates. However, the possibility of residual confounding remains. Although a large randomised controlled trial could overcome this limitation, a previous study suggested that such a trial is unlikely to be feasible.Our study only included children who could be linked to pupil census records, with likely explanations for any gaps in linking including that the child had emigrated from Scotland or had attended private school (around 4% of children in Scotland attend private schools and are not included in the pupil census).To our knowledge, to date only one study has investigated the effect of planned VBAC compared with ERCS on child neurodevelopmental outcomes.It is possible that the slight increase in risk of any SENs and the increased risk of sensory impairment seen for planned VBAC with labour induction may be related to the increased risk of complications, most notably uterine rupture, that has been observed, particularly with inducing labour in women with previous caesarean section.This study provides little evidence of an association between planned mode of birth after previous caesarean section and SENs beyond a small absolute increased risk of sensory impairment seen for planned VBAC with labour induction. This finding may be the result of performing multiple comparisons or residual confounding. The findings provide valuable additional information to manage and counsel women with previous caesarean section concerning their future birth choices, but further research is needed to establish whether the results can be replicated and to explore the impact of planned mode of birth after previous caesarean section on other measures of child neurodevelopmental adversity.KEF received financial support from the National Institute for Health Research (NIHR) for the submitted work. Completed disclosure of interests form available to view online as supporting information. The other authors reported none.KEF conceived the study, gained funding and approvals for the study, cleaned and analysed the data and wrote the first draft of the article. MAQ and JJK supervised the study. All authors contributed to the design and interpretation of the study and critically reviewed the article and approved the final version for publication.The study was exempt from UK National Research Ethics Service approval as it involved secondary analysis of anonymised data. However, study approval was obtained from the Public Benefit and Privacy Panel for Health and Social Care Scotland (1516‐0196) and the Scottish Government Education Analytical Services.KEF was funded by a National Institute for Health Research (NIHR) Doctoral Research Fellowship (DRF‐2016‐09‐078) for this research project. This paper presents independent research. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the article.The authors would like to acknowledge the support of the eDRIS Team  for their involvement in obtaining approvals and provisioning and linking data, and the use of the secure analytical platform within the National Safe Haven.https://www.isdscotland.org/Products‐and‐Services/EDRIS/ and at https://www.gov.scot/publications/scottish‐government‐statistics‐request‐our‐data/.Data (eDRIS study number 1516‐0196) are available from the Information Services Division Scotland, now part of Public Health Scotland, following the necessary approval process by the Public Benefit and Privacy Panel for Health and Social Care and the Scottish Government Education Analytical Services. More details can be found at Figure S1. Conception framework of how sociodemographic, maternal medical and pregnancy‐related and infant‐related factors might influence the relationship between planned mode of birth after previous caesarean and special educational needs (SENs) in the child.Figure S2. Outcomes following: (A) planned VBAC, compared with ERCS; (B) planned VBAC without labour induction, compared with ERCS; (C) planned VBAC with labour induction, compared with ERCS; (D) planned and actual VBAC, compared with ERCS; and (E) planned VBAC but in‐labour non‐elective repeat caesarean section, compared with ERCS.Table S1. Data sources.Table S2. Data sources, codes and database fields used to identify study population, exposures, outcomes and covariates.Table S3. Characteristics of children included in the study compared with those not included because they did not link to any pupil census records between 2007 and 2016 when the child was aged 4–11 years and attending a primary or special school.Table S4. Outcomes according to actual mode of birth: planned and actually had VBAC and planned VBAC but had in‐labour non‐elective repeat caesarean section, compared with ERCS.Table S5. Complete case analysis of outcomes following planned VBAC, compared with ERCS.Table S6. Complete case analysis of outcomes following planned VBAC with and without labour induction, compared with ERCS.Table S7. Complete case analysis of outcomes according to actual mode of birth: planned and actual VBAC and planned VBAC but in‐labour non‐elective repeat caesarean section, compared with ERCS.Table S8. Outcomes following planned VBAC compared with ERCS at ≥ 39 weeks of gestation.Table S9. Outcomes following planned VBAC with and without labour induction, compared with ERCS, at ≥39 weeks of gestation.Table S10. Outcomes according to actual mode of birth: planned and actual VBAC and planned VBAC but in‐labour non‐elective repeat caesarean section, compared with ERCS, at ≥39 weeks of gestation.Table S11. Outcomes following planned VBAC compared with ERCS, analysing SENs as a repeated‐measures yearly outcome.Table S12. Outcomes following planned VBAC with and without labour induction compared with ERCS, analysing SENs as a repeated‐measures yearly outcome.Table S13. Outcomes according to actual mode of birth: planned and actual VBAC and planned VBAC but in‐labour non‐elective repeat caesarean section, compared with ERCS, analysing SENs as a repeated‐measures yearly outcome.Table S14. Outcomes following planned VBAC compared with ERCS, removing covariate maternal BMI from the analysis.Table S15. Outcomes following planned VBAC with and without labour induction, compared with ERCS, removing covariate maternal BMI from the analysis.Table S16. Outcomes according to actual mode of birth: planned and actual VBAC and planned VBAC but in‐labour non‐elective repeat caesarean section, compared with ERCS, removing covariate maternal BMI from the analysis.Click here for additional data file.Supplementary MaterialClick here for additional data file.Supplementary MaterialClick here for additional data file.Supplementary MaterialClick here for additional data file.