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  • Exposure to high altitude is associated with an elevated risk of hip fracture: a retrospective cohort study using data from the CHARLS | BMC Geriatrics

    Exposure to high altitude is associated with an elevated risk of hip fracture: a retrospective cohort study using data from the CHARLS | BMC Geriatrics

    Study population and dataset

    The CHARLS is a prospective national cohort study that enrolled 17,708 participants in 2011, and three waves of follow-up were conducted in 2013, 2015, and 2018. Participants were randomly selected using a probability-proportional-to-size (PPS) technique and a four-stage random sample method. The workgroup selected 150 counties in 28 provinces. Administrative villages in rural areas and neighborhoods in urban areas were the primary sampling units (PSUs). Three PSUs within each county-level unit were selected using PPS sampling. Detailed information on the methodology and cohort profile has been reported previously [11].

    We collected data on participants enrolled in the baseline investigation who attended all three follow-up investigations and aged above 60 years old. Those with a history of cancer were excluded because the progression and treatment of cancer affect multiple organs and systems throughout the body; this aspect could have introduced substantial bias into our study. During the data cleaning process, we identified participants with implausible data regarding body weight and height, specifically heights less than 100 cm or weights less than 20 kg. These values were considered erroneous owing to likely data entry mistakes; therefore, participants with an abnormal body mass index (BMI) (BMI < 10 or BMI > 60) in the baseline investigation were excluded.

    Altitude data acquisition

    The participants’ place of residence was determined by their community ID. The latitude and longitude were acquired through amap api (https://restapi.amap.comv3/geocode), and the local altitude was acquired based on the latitude and longitude using geodata (version 0.5-8) [12] packages and the raster package (version 3.6–23) [13]. Participants were stratified into a low-altitude or high-altitude group based on a criterion of 1500 m, which was used in previous studies [14].

    Variable collection

    The primary outcome was hip fractures reported by the participants, which was defined by their answer to the question, “Have you fractured your hip since the last interview?” Participants choosing “yes” were considered to have experienced a hip fracture, and the time point the investigation occurred was recorded as the time the event happened.

    Individual income, marital status, medical history, sex, BMI, smoking status, alcohol consumption, and age were selected as covariates for propensity score matching (PSM). Individual income was acquired from harmonized CHARLS data and evenly divided into five groups. Educational status was retrieved from the answer to the question: “What is the highest level of education completed?” and re-coded into preschool, primary, secondary, and higher education. Marital status was retrieved from the answer to the question: “What is your marital status?” and re-coded as unmarried, married, widowed, divorced (or long-term separation). History of stroke, cardiovascular dysfunction, falls, hip fractures, chronic lung diseases, and arthritis were retrieved from the answer to the question: “Have you been diagnosed with any of the following by a doctor?” The presence of specific diseases was determined based on whether the participant selected “Yes” to the following conditions: (1) “Chronic lung diseases, such as chronic bronchitis and emphysema (excluding tumors or cancer),” (2) “Heart attack, coronary heart disease, angina, congestive heart failure, or other heart problems,” (3) “Stroke,” and (4) “Arthritis or rheumatism.” BMI was calculated using the following formula: BMI (kg/m[2]) = body weight (kg)/body height (m)2.

    Propensity score matching

    Individual income, marital status, medical history, sex, BMI, smoking status, alcohol consumption, and age were selected for PSM. A general linear model was used to calculate the distance, and the nearest method was used for matching with a matching ratio of 1:8 (high altitude: low altitude). After matching, a balance test was performed to evaluate the imbalance between the two groups in the matched data. The MatchIt package [15] was used for matching, and the Cobalt package [16] was used for the balanced test and plotting.

    Statistical analysis

    Categorical variables are shown as counts (percentages, %), and continuous variables are shown as mean ± SD. To compare the differences in baseline data between the two groups, chi-square and t-tests were used for categorical and continuous variables, respectively. Kaplan–Meier survival analysis and Cox regression were used to compare the differences between the two groups regarding fall and hip fracture risks.

    Subgroup analysis was performed, and the participants were divided into subgroups according to sex, smoking history, and overweight status. The effects of high-altitude exposure on fall and hip fracture risks in different subgroups were evaluated using Cox regression analysis.

    To test the robustness of our results, we performed different sensitivity analyses as follows: (1) We randomly sampled 80% and 90% of all participants, respectively, and replicated the previously described analysis workflow. (2) Using optimal matching as the PSM method and a matching ratio of 1:8, we replicated the previously described analysis workflow. (3) Cox regression was performed with and without adjustments for covariates without matching the participants.

    All analyses were performed using R (version 4.3.1). The survival analysis was performed using the survival package (version 3.5-5) [17] and visualized using the Survminer package (version 0.4.9) [18]. The comparison of baseline characteristics and the generation of tables was performed using the gtsummary package (version 1.7.2) [19].

    Continue Reading

  • World Health Organization. Guidelines for the psychosocially assisted Pharmacological treatment of opioid dependence. Geneva, Switzerland: World Health Organization; 2009. p. 9241547545. Report No.

    Google Scholar 

  • Bruneau J, Ahamad K, Goyer M-È, Poulin G, Selby P, Fischer B, et al. Management of opioid use disorders: A National clinical practice guideline. Can Med Assoc J. 2018;190(9):E247–57.

    Article 

    Google Scholar 

  • Priest KC, Gorfinkel L, Klimas J, Jones AA, Fairbairn N, McCarty D. Comparing Canadian and united States opioid agonist therapy policies. Int J Drug Policy. 2019;74:257–65.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Eibl JK, Morin K, Leinonen E, Marsh DC. The state of opioid agonist therapy in Canada 20 years after federal oversight. Can J Psychiatry Revue Canadienne De Psychiatrie. 2017;62(7):444–50.

    Article 
    PubMed 

    Google Scholar 

  • Piske M, Zhou H, Min JE, Hongdilokkul N, Pearce LA, Homayra F, et al. The cascade of care for opioid use disorder: A retrospective study in British Columbia, Canada. Addiction. 2020;115(8):1482–93.

    Article 
    PubMed 

    Google Scholar 

  • Tahsin F, Morin KA, Vojtesek F, Marsh DC. Measuring treatment attrition at various stages of engagement in opioid agonist treatment in Ontario Canada using a cascade of care framework. BMC Health Serv Res. 2022;22(1):490.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • O’Connor AM, Cousins G, Durand L, Barry J, Boland F. Retention of patients in opioid substitution treatment: A systematic review. PLoS ONE. 2020;15(5):e0232086.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Fischer B. The continuous opioid death crisis in canada: changing characteristics and implications for path options forward. Lancet Reg Health – Americas. 2023;19:100437.

    Article 
    PubMed 

    Google Scholar 

  • Sordo L, Barrio G, Bravo MJ, Indave BI, Degenhardt L, Wiessing L, et al. Mortality risk during and after opioid substitution treatment: systematic review and meta-analysis of cohort studies. BMJ. 2017;357:j1550.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Hall NY, Le L, Majmudar I, Mihalopoulos C. Barriers to accessing opioid substitution treatment for opioid use disorder: A systematic review from the client perspective. Drug Alcohol Depend. 2021;221:108651.

    Article 
    PubMed 

    Google Scholar 

  • Lachapelle É, Archambault L, Blouin C, Perreault M. Perspectives of people with opioid use disorder on improving addiction treatments and services. Drugs: Educ Prev Policy. 2021;28(4):316–27.

    Google Scholar 

  • Pearce LA, Min JE, Piske M, Zhou H, Homayra F, Slaunwhite A, et al. Opioid agonist treatment and risk of mortality during opioid overdose public health emergency: population based retrospective cohort study. BMJ. 2020;368:m772.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Mackay L, Kerr T, Fairbairn N, Grant C, Milloy MJ, Hayashi K. The relationship between opioid agonist therapy satisfaction and Fentanyl exposure in a Canadian setting. Addict Sci Clin Pract. 2021;16(1):26.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Mead N, Bower P. Patient-centredness: a conceptual framework and review of the empirical literature. Soc Sci Med. 2000;51(7):1087–110.

    Article 
    PubMed 

    Google Scholar 

  • Greene SM, Tuzzio L, Cherkin D. A framework for making patient-centered care front and center. Permanente J. 2012;16(3):49.

    Article 

    Google Scholar 

  • Kitson A, Marshall A, Bassett K, Zeitz K. What are the core elements of patient-centred care? A narrative review and synthesis of the literature from health policy, medicine and nursing. J Adv Nurs. 2012;69(1):4–15.

    Article 
    PubMed 

    Google Scholar 

  • Scholl I, Zill JM, Härter M, Dirmaier J. An integrative model of patient-centeredness – A systematic review and concept analysis. PLoS ONE. 2014;9(9):e107828.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Marchand K, Beaumont S, Westfall J, MacDonald S, Harrison S, Marsh DC, et al. Conceptualizing patient-centered care for substance use disorder treatment: findings from a systematic scoping review. Subst Abuse Treat Prev Policy. 2019;14(1):37.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Brothers TD, Bonn M. Patient-centred care in opioid agonist treatment could improve outcomes. Can Med Assoc J. 2019;191(17):E460–1.

    Article 

    Google Scholar 

  • British Columbia Centre on Substance Use BMoH, and BC Ministry of Mental Health and Addictions. A guideline for the clinical management of opioid use disorder. Vancouver, BC: British Columbia Centre on Substance Use (BCCSU); 2023.

  • Rastegar DA. Patient-Centered care in opioid use disorder treatment. In: Wakeman SE, Rich JD, editors. Treating opioid use disorder in general medical settings. Cham: Springer International Publishing; 2021. pp. 1–7.

    Google Scholar 

  • Edland-Gryt M, Skatvedt AH. Thresholds in a low-threshold setting: an empirical study of barriers in a centre for people with drug problems and mental health disorders. Int J Drug Policy. 2013;24(3):257–64.

    Article 
    PubMed 

    Google Scholar 

  • Biancarelli DL, Biello KB, Childs E, Drainoni M, Salhaney P, Edeza A, et al. Strategies used by people who inject drugs to avoid stigma in healthcare settings. Drug Alcohol Depend. 2019;198:80–6.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Harris M, Rhodes T, Martin A. Taming systems to create enabling environments for HCV treatment: negotiating trust in the drug and alcohol setting. Soc Sci Med. 2013;83:19–26.

    Article 
    PubMed 

    Google Scholar 

  • Treloar C, Rance J, Yates K, Mao L. Trust and people who inject drugs: the perspectives of clients and staff of needle syringe programs. Int J Drug Policy. 2016;27:138–45.

    Article 
    PubMed 

    Google Scholar 

  • Farrugia A, Pienaar K, Fraser S, Edwards M, Madden A. Basic care as exceptional care: addiction stigma and consumer accounts of quality healthcare in Australia. Health Sociol Rev. 2021;30(2):95–110.

    Article 
    PubMed 

    Google Scholar 

  • Fraser S, Moore D, Farrugia A, Edwards M, Madden A. Exclusion and hospitality: the subtle dynamics of stigma in healthcare access for people emerging from alcohol and other drug treatment. Sociol Health Illn. 2020;42(8):1801–20.

    Article 
    PubMed 

    Google Scholar 

  • Muncan B, Walters SM, Ezell J, Ompad DC. They look at Us like junkies: influences of drug Use stigma on the healthcare engagement of people who inject drugs in new York City. Harm Reduct J. 2020;17(1):53.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Addison M, Lhussier M, Bambra C. Relational stigma as a social determinant of health: i’m not what you _____see me as. SSM – Qualitative Res Health. 2023;4:100295.

    Article 

    Google Scholar 

  • Marshall K, Maina G, Sherstobitoff J. Plausibility of patient-centred care in high-intensity methadone treatment: reflections of providers and patients. Addict Sci Clin Pract. 2021;16(1):42.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Treloar C, Rance J, Backmund M. Understanding barriers to hepatitis C virus care and stigmatization from a social perspective. Clin Infect Dis. 2013;57(Suppl 2):S51–5.

    Article 
    PubMed 

    Google Scholar 

  • Heller D, McCoy K, Cunningham C. An invisible barrier to integrating HIV primary care with harm reduction services: philosophical clashes between the harm reduction and medical models. Public Health Rep. 2004;119(1):32–9.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Denis-Lalonde D, Lind C, Estefan A. Beyond the buzzword: A concept analysis of harm reduction. Res Theory Nurs Pract. 2019;4:310–23.

    Article 

    Google Scholar 

  • Frankeberger J, Gagnon K, Withers J, Hawk M. Harm reduction principles in a street medicine program: A qualitative study. Culture. Med Psychiatry. 2023;47(4):1005–21.

    Article 

    Google Scholar 

  • McNeil R, Kerr T, Pauly B, Wood E, Small W. Advancing patient-centered care for structurally vulnerable drug-using populations: A qualitative study of the perspectives of people who use drugs regarding the potential integration of harm reduction interventions into hospitals. Addiction. 2016;111(4):685–94.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • McNeil R, Small W. Safer environment interventions’: A qualitative synthesis of the experiences and perceptions of people who inject drugs. Soc Sci Med. 2014;106:151–8.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Jakubowski A, Rath C, Harocopos A, Wright M, Welch A, Kattan J, et al. Implementation of buprenorphine services in NYC syringe services programs: A qualitative process evaluation. Harm Reduct J. 2022;19(1):75.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Latour B, Venn C. Morality and Technology. Theory. Cult Soc. 2002;19(5–6):247–60.

    Article 

    Google Scholar 

  • Fomiatti R, Shaw F, Fraser S. It’s a different way to do medicine’: exploring the affordances of telehealth for hepatitis C healthcare. Int J Drug Policy. 2022;110:103875.

    Article 
    PubMed 

    Google Scholar 

  • Høj SB, de Montigny C, Chougar S, Léandre R, Beauchemin-Nadeau M-È, Boyer-Legault G, et al. Co-constructing a community-based telemedicine program for people with opioid use disorder during the COVID-19 pandemic: lessons learned and implications for future service delivery. JMIR Public Health Surveillance. 2023;9:e39236.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Jauffret-Roustide M, Bertrand K. COVID-19, usages de drogues et réduction des risques: analyse croisée des expériences et de l’impact de La pandémie En France et Au Québec. Criminologie. 2022;55(2):17–42.

    Article 

    Google Scholar 

  • Minoyan N, Høj SB, Zolopa C, Vlad D, Bruneau J, Larney S. Self-reported impacts of the COVID-19 pandemic among people who use drugs: A rapid assessment study in Montreal, Canada. Harm Reduct J. 2022;19(1):38.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Zolopa C, Brothers TD, Leclerc P, Mary J-F, Morissette C, Bruneau J, et al. Changes in supervised consumption site use and emergency interventions in Montréal, Canada in the first twelve months of the COVID-19 pandemic: an interrupted time series study. Int J Drug Policy. 2022;110:103894.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Ali F, Russell C, Nafeh F, Rehm J, LeBlanc S, Elton-Marshall T. Changes in substance supply and use characteristics among people who use drugs (PWUD) during the COVID-19 global pandemic: A National qualitative assessment in Canada. Int J Drug Policy. 2021;93:103237.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Russell C, Ali F, Nafeh F, Rehm J, LeBlanc S, Elton-Marshall T. Identifying the impacts of the COVID-19 pandemic on service access for people who use drugs (PWUD): A National qualitative study. J Subst Abuse Treat. 2021;129:108374.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Imtiaz S, Nafeh F, Russell C, Ali F, Elton-Marshall T, Rehm J. The impact of the novel coronavirus disease (COVID-19) pandemic on drug overdose-related deaths in the United States and Canada: a systematic review of observational studies and analysis of public health surveillance data. Substance Abuse Treat Prevent Policy. 2021;16(1):87.

  • Krawczyk N, Fawole A, Yang J, Tofighi B. Early innovations in opioid use disorder treatment and harm reduction during the COVID-19 pandemic: a scoping review. Addict Sci Clin Pract. 2021;16(1):68.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Alami H, Lehoux P, Attieh R, Fortin J-P, Fleet R, Niang M, et al. A not so quiet revolution: systemic benefits and challenges of telehealth in the context of COVID-19 in Quebec (Canada). Front Digit Health. 2021;3:721898.

    Article 

    Google Scholar 

  • Glegg S, McCrae K, Kolla G, Touesnard N, Turnbull J, Brothers TD, et al. COVID just kind of opened a can of whoop-ass: the rapid growth of safer supply prescribing during the pandemic documented through an environmental scan of addiction and harm reduction services in Canada. Int J Drug Policy. 2022;106:103742.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Letter from the Minister of Health regarding treatment and safer supply [press release]. Ottawa, ON: Government of Canada, 24 August 2020.

  • Brar R, Bruneau J, Butt P, Goyer M, Lim R, Poulin G, et al. Medications and other clinical approaches to support physical distancing for people who use substances during the COVID-19 pandemic: National rapid guidance document. Vancouver, BC: Canadian Research Initiative in Substance Misuse; 2020.

  • Bruneau J, Rehm J, Wild TC, Wood E, Sako A, Swansburg J, Lam A. Telemedicine support for addiction services: National rapid guidance document. Montreal, QC: Canadian Research Initiative in Substance Misuse; 2020.

    Google Scholar 

  • Health Canada. Subsection 56(1) class exemption for patients, practitioners and pharmacists prescribing and providing controlled substances in Canada Ottawa, ON: Government of Canada. 2020. https://www.canada.ca/en/health-canada/services/health-concerns/controlled-substances-precursor-chemicals/policy-regulations/policy-documents/section-56-1-class-exemption-patients-pharmacists-practitioners-controlled-substances-covid-19-pandemic.html. Accessed 1 Nov 2021.

  • Goyer M-È, Hudon K, Plessis-Bélair M-C, Ferguson Y. Substance replacement therapy in the context of the COVID-19 pandemic in Québec: clinical guidance for prescribers. Montreal, QC: Institut universitaire Sur les dépendances (IUD); 2020. https://dependanceitinerance.ca/app/uploads/2020/10/Guide-Pharmaco-COVID_ANG-VF.19.10.20.pdf. Accessed 10 Oct 2023.

  • Collège des médecins du Québec. Trouble lié à l’utilisation d’opioïdes (TUO): Prescription d’un traitement par agonistes opioïdes (TAO) durant la pandémie Montreal, QC: Collège des médecins du Québec. 2020. http://www.cmq.org/page/fr/covid-19-trouble-lie-a-l-utilisation-d-opioides-tuo-prescription-d-un-traitement-par-agonistes-opioides-tao-durant-la-pandemie.aspx. Accessed 8 Apr 2020.

  • Collège des médecins du Québec. Les téléconsultations réalisées par les médecins durant la pandémie de COVID-19: Guide à l’intention des médecins. Montreal, QC: Collège des médecins du Québec; 31 March 2020. Accessed 11 May 2020.

  • Mignacca FG. Montreal’s CACTUS safe-injecting community group sees more overdoses amid pandemic Montreal, QC: CBC News; 2020. Accessed 19 Jun 2024. https://www.cbc.ca/news/canada/montreal/cactus-montreal-opioid-overdoses-covid-19-1.5618901

  • Nosyk B, Min JE, Pearce LA, Zhou H, Homayra F, Wang L, et al. Development and validation of health system performance measures for opioid use disorder in British Columbia, Canada. Drug Alcohol Depend. 2022;233:109375.

    Article 
    PubMed 

    Google Scholar 

  • Kurz M, Min JE, Dale LM, Nosyk B. Assessing the determinants of completing OAT induction and long-term retention: A population-based study in British Columbia, Canada. J Subst Abuse Treat. 2022;133:108647.

    Article 
    PubMed 

    Google Scholar 

  • Krebs E, Homayra F, Min JE, MacDonald S, Gold L, Carter C, Nosyk B. Characterizing opioid agonist treatment discontinuation trends in British Columbia, Canada, 2012–2018. Drug Alcohol Depend. 2021;225:108799.

    Article 
    PubMed 

    Google Scholar 

  • Nielsen S, Degenhardt L, Hoban B, Gisev N. A synthesis of oral morphine equivalents (OME) for opioid utilisation studies. Pharmacoepidemiol Drug Saf. 2016;25(6):733–7.

  • Patterson Silver Wolf DA, Gold M. Treatment resistant opioid use disorder (TROUD): Definition, rationale, and recommendations. J Neurol Sci. 2020;411:116718.

    Article 
    PubMed 

    Google Scholar 

  • Kimmel S, Bach P, Walley AY. Comparison of treatment options for refractory opioid use disorder in the united States and canada: A narrative review. J Gen Intern Med. 2020;35(8):2418–26.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Voon P, Joe R, Fairgrieve C, Ahamad K. Treatment of opioid use disorder in an innovative community-based setting after multiple treatment attempts in a woman with untreated HIV. BMJ Case Rep. 2016;2016:bcr2016215557.

  • Håkansson Eklund J, Holmström IK, Kumlin T, Kaminsky E, Skoglund K, Höglander J, et al. Same same or different? A review of reviews of person-centered and patient-centered care. Patient Educ Couns. 2019;102(1):3–11.

    Article 
    PubMed 

    Google Scholar 

  • Marlatt GA, Blume AW, Parks GA. Integrating harm reduction therapy and traditional substance abuse treatment. J Psychoactive Drugs. 2001;33(1):13–21.

    Article 
    PubMed 

    Google Scholar 

  • Chang JE, Lindenfeld Z, Hagan H. Integrating harm reduction into medical care: lessons from three models. J Am Board Fam Med. 2023;36(3):449–61.

    Article 
    PubMed 

    Google Scholar 

  • Salvalaggio G, McKim R, Taylor M, Wild TC. Patient–provider rapport in the health care of people who inject drugs. SAGE Open. 2013;3(4):2158244013509252.

    Article 

    Google Scholar 

  • Lilly R, Quirk A, Rhodes T, Stimson GV. Sociality in methadone treatment: Understanding methadone treatment and service delivery as a social process. Drugs: Educ Prev Policy. 2000;7(2):163–78.

    Google Scholar 

  • Hall MA, Dugan E, Zheng B, Mishra AK. Trust in physicians and medical institutions: what is it, can it be measured, and does it matter? Milbank Q. 2001;79(4):613–39.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Morin KA, Tatangelo M, Marsh D. Canadian addiction treatment centre (CATC) opioid agonist treatment cohort in Ontario, Canada. BMJ Open. 2024;14(2):e080790.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Vogel M, Dürsteler KM, Walter M, Herdener M, Nordt C. Rethinking retention in treatment of opioid dependence: the eye of the beholder. Int J Drug Policy. 2017;39:109–13.

    Article 
    PubMed 

    Google Scholar 

  • Santo T Jr, Clark B, Hickman M, Grebely J, Campbell G, Sordo L, et al. Association of opioid agonist treatment with All-Cause mortality and specific causes of death among people with opioid dependence: A systematic review and Meta-analysis. JAMA Psychiatry. 2021;78(9):979–93.

    Article 
    PubMed 

    Google Scholar 

  • Nosyk B, MacNab YC, Sun H, Fischer B, Marsh DC, Schechter MT, Anis AH. Proportional hazards frailty models for recurrent methadone maintenance treatment. Am J Epidemiol. 2009;170(6):783–92.

    Article 
    PubMed 

    Google Scholar 

  • Pilarinos A, Kwa Y, Joe R, Thulien M, Buxton JA, DeBeck K, Fast D. Navigating opioid agonist therapy among young people who use illicit opioids in Vancouver, Canada. Int J Drug Policy. 2022;107:103773.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Bell J, Burrell T, Indig D, Gilmour S. Cycling in and out of treatment: participation in methadone treatment in NSW, 1990–2002. Drug Alcohol Depend. 2006;81(1):55–61.

    Article 
    PubMed 

    Google Scholar 

  • Nordt C, Vogel M, Dey M, Moldovanyi A, Beck T, Berthel T, et al. One size does not fit all—evolution of opioid agonist treatments in a naturalistic setting over 23 years. Addiction. 2019;114(1):103–11.

    Article 
    PubMed 

    Google Scholar 

  • Muller AE, Bjørnestad R, Clausen T. Dissatisfaction with opioid maintenance treatment partly explains reported side effects of medications. Drug Alcohol Depend. 2018;187:22–8.

    Article 
    PubMed 

    Google Scholar 

  • Giang K, Charlesworth R, Thulien M, Mulholland A, Barker B, Brar R, et al. Risk mitigation guidance and safer supply prescribing among young people who use drugs in the context of COVID-19 and overdose emergencies. Int J Drug Policy. 2023;115:104023.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Min JE, Guerra-Alejos BC, Yan R, Palis H, Barker B, Urbanoski K, et al. Opioid coprescription through risk mitigation guidance and opioid agonist treatment receipt. JAMA Netw Open. 2024;7(5):e2411389–e.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Sanders JJ, Roose RJ, Lubrano MC, Lucan SC. Meaning and methadone: patient perceptions of methadone dose and a model to promote adherence to maintenance treatment. J Addict Med. 2013;7(5):307–13.

  • Roux P, Lions C, Michel L, Cohen J, Mora M, Marcellin F, et al. Predictors of non-adherence to methadone maintenance treatment in opioid-dependent individuals: implications for clinicians. Curr Pharm Design. 2014;20(25):4097–105.

    Article 

    Google Scholar 

  • Artenie AA, Minoyan N, Jacka B, Høj S, Jutras-Aswad D, Roy É, et al. Opioid agonist treatment dosage and patient-perceived dosage adequacy, and risk of hepatitis C infection among people who inject drugs. Can Med Assoc J. 2019;191(17):E462–8.

    Article 

    Google Scholar 

  • Hämmig R, Köhler W, Bonorden-Kleij K, Weber B, Lebentrau K, Berthel T, et al. Safety and tolerability of slow-release oral morphine versus methadone in the treatment of opioid dependence. J Subst Abuse Treat. 2014;47(4):275–81.

    Article 
    PubMed 

    Google Scholar 

  • Kastelic A, Dubajic G, Strbad E. Slow-release oral morphine for maintenance treatment of opioid addicts intolerant to methadone or with inadequate withdrawal suppression. Addiction. 2008;103(11):1837–46.

    Article 
    PubMed 

    Google Scholar 

  • Yarborough BJH, Stumbo SP, McCarty D, Mertens J, Weisner C, Green CA. Methadone, buprenorphine and preferences for opioid agonist treatment: A qualitative analysis. Drug Alcohol Depend. 2016;160:112–8.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Mocanu V, Bozinoff N, Wood E, Jutras-Aswad D, Le Foll B, Lim R, et al. Opioid agonist therapy switching among individuals with prescription-type opioid use disorder: secondary analysis of a pragmatic randomized trial. Drug Alcohol Depend. 2023;248:109932.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Saxon AJ. Short-acting, full agonist opioids during initiation of opioid agonist treatment in the Fentanyl era. JAMA Netw Open. 2024;7(5):e2411398–e.

    Article 
    PubMed 

    Google Scholar 

  • Behnia B. Trust development: A discussion of three approaches and a proposed alternative. Br J Social Work. 2007;38(7):1425–41.

    Article 

    Google Scholar 

  • Fine GA. Agency, structure, and comparative contexts: toward a synthetic interactionism. Symbolic Interact. 1992;15(1):87–107.

    Article 

    Google Scholar 

  • Rance J, Newland J, Hopwood M, Treloar C. The politics of place(ment): problematising the provision of hepatitis C treatment within opiate substitution clinics. Soc Sci Med. 2012;74(2):245–53.

    Article 
    PubMed 

    Google Scholar 

  • Harris J, McElrath K. Methadone as social control: institutionalized stigma and the prospect of recovery. Qual Health Res. 2012;22(6):810–24.

    Article 
    PubMed 

    Google Scholar 

  • Harris M, Albers E, Swan T. The promise of treatment as prevention for hepatitis C: meeting the needs of people who inject drugs? Int J Drug Policy. 2015;26(10):963–9.

    Article 
    PubMed 

    Google Scholar 

  • Hall MA, Camacho F, Dugan E, Balkrishnan R. Trust in the medical profession: conceptual and measurement issues. Health Serv Res. 2002;37(5):1419–39.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Aung PTZ, Spelman T, Wilkinson AL, Dietze PM, Stoové MA, Hellard ME. Time-to-hepatitis C treatment initiation among people who inject drugs in Melbourne, Australia. Epidemiol Infect. 2023;151:e84.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Jiang N, Bruneau J, Makarenko I, Minoyan N, Zang G, Høj SB, et al. HCV treatment initiation in the era of universal direct acting antiviral coverage – Improvements in access and persistent barriers. Int J Drug Policy. 2023;113:103954.

    Article 
    PubMed 

    Google Scholar 

  • Malme KB, Ulstein K, Finbråten A-K, Wüsthoff LEC, Kielland KB, Hauge J, et al. Hepatitis C treatment uptake among people who inject drugs in Oslo, norway: A registry-based study. Int J Drug Policy. 2023;116:104044.

    Article 
    PubMed 

    Google Scholar 

  • Papalamprakopoulou Z, Ntagianta E, Triantafyllou V, Kalamitsis G, Dharia A, Dickerson SS, et al. Breaking the vicious cycle of delayed healthcare seeking for people who use drugs. Harm Reduct J. 2025;22(1):27.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Cochran G, Bruneau J, Cox N, Gordon AJ. Medication treatment for opioid use disorder and community pharmacy: expanding care during a National epidemic and global pandemic. Subst Abus. 2020;41(3):269–74.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Fatani S, Bakke D, D’Eon M, El-Aneed A. Qualitative assessment of patients’ perspectives and needs from community pharmacists in substance use disorder management. Subst Abuse Treat Prevent Policy. 2021;16(1):38.

  • Bishop LD, Rosenberg-Yunger ZRS. Pharmacists expanded role in providing care for opioid use disorder during COVID-19: A qualitative study exploring pharmacists’ experiences. Drug Alcohol Depend. 2022;232:109303.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Tyndall M. Safer opioid distribution in response to the COVID-19 pandemic. Int J Drug Policy. 2020;83:102880.

  • Bardwell G, Ivsins A, Wallace JR, Mansoor M, Kerr T. The machine doesn’t judge: counternarratives on surveillance among people accessing a safer opioid supply via biometric machines. Soc Sci Med. 2024;345:116683.

    Article 
    PubMed 

    Google Scholar 

Continue Reading

  • Eating disorders symptoms and associated risk factors among medical students in France: a nationwide cross-sectional study | Journal of Eating Disorders

    Eating disorders symptoms and associated risk factors among medical students in France: a nationwide cross-sectional study | Journal of Eating Disorders

  • Rotenstein LS, Ramos MA, Torre M, Segal JB, Peluso MJ, Guille C, et al. Prevalence of depression, depressive symptoms, and suicidal ideation among medical students: a systematic review and meta-analysis. JAMA. 2016;316(21):2214–36. https://doi.org/10.1001/jama.2016.17324.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Wang J, Fitzke RE, Tran DD, Grell J, Pedersen ER. Mental health treatment-seeking behaviors in medical students: a mixed-methods approach. J Med Access. 2023;15(7):27550834221147788. https://doi.org/10.1177/27550834221147787.

    Article 

    Google Scholar 

  • Quek TT, Tam WW, Tran BX, Zhang M, Zhang Z, Ho CS, et al. The global prevalence of anxiety among medical students: a meta-analysis. Int J Environ Res Public Health. 2019;16(15):2735. https://doi.org/10.3390/ijerph16152735.

    Article 
    PubMed 

    Google Scholar 

  • Tavolacci MP, Ladner J, Dechelotte P. COVID-19 pandemic and eating disorders among university students. Nutrients. 2021;13(12):4294. https://doi.org/10.3390/nu13124294.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Pacanowski CR, Skubisz C, Borton D, Ryding R. Prevalence and correlates of disordered eating at a large state university before and after the onset of the COVID-19 pandemic. J Eat Disord. 2024;12(1):153. https://doi.org/10.1186/s40337-024-01056-2.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Galmiche M, Déchelotte P, Lambert G, Tavolacci MP. Prevalence of eating disorders over the 2000–2018 period: a systematic literature review. Am J Clin Nutr. 2019;109(5):1402–13. https://doi.org/10.1093/ajcn/nqy342.

    Article 
    PubMed 

    Google Scholar 

  • Andreeva VA, Tavolacci MP, Galan P, Ladner J, Buscail C, Péneau S, et al. Sociodemographic correlates of eating disorder subtypes among men and women in France, with a focus on age. J Epidemiol Community Health. 2019;73(1):56–64. https://doi.org/10.1136/jech-2018-210745.

    Article 
    PubMed 

    Google Scholar 

  • Fekih-Romdhane F, Daher-Nashif S, Alhuwailah AH, Al Gahtani HMS, Hubail SA, Shuwiekh HAM, Khudhair MF, Alhaj OA, Bragazzi NL, Jahrami H. The prevalence of feeding and eating disorders symptomology in medical students: an updated systematic review, meta-analysis, and meta-regression. Eat Weight Disord. 2022 Aug;27(6):1991–2010. https://doi.org/10.1007/s40519-021-01351-w.

  • Rolland F, Hadouiri N, Haas-Jordache A, Gouy E, Mathieu L, Goulard A, Morvan Y, Frajerman A. Mental health and working conditions among French medical students: A nationwide study. J Affect Disord. 2022 Jun 1;306:124–130. https://doi.org/10.1016/j.jad.2022.03.001. Epub 2022 Mar 8. Erratum in: J Affect Disord. 2022 Jul 1;308:623–624. https://doi.org/10.1016/j.jad.2022.04.017. Erratum in: J Affect Disord. 2023 Feb 15;323:901–902. https://doi.org/10.1016/j.jad.2022.12.046.

  • Munn-Chernoff MA, Few LR, Matherne CE, Baker JH, Men VY, McCutcheon VV, Agrawal A, Bucholz KK, Madden PAF, Heath AC, Duncan AE. Eating disorders in a community-based sample of women with alcohol use disorder and nicotine dependence. Drug Alcohol Depend. 2020 Jul 1;212:107981. https://doi.org/10.1016/j.drugalcdep.2020.107981. Epub 2020 Apr 25.

  • Galmiche M, Godefroy C, Achamrah N, Grigioni S, Colange G, Folope V, et al. Mental health and health behaviours among patients with eating disorders: a case-control study in France. J Eat Disord. 2022;10(1):160. https://doi.org/10.1186/s40337-022-00691-x.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Lamberti M, Napolitano F, Napolitano P, Arnese A, Crispino V, Panariello G, et al. Prevalence of alcohol use disorders among under- and post-graduate healthcare students in Italy. PLoS ONE. 2017;12:e0175719.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Tavolacci MP, Delay J, Grigioni S, Déchelotte P, Ladner J. Changes and specificities in health behaviors among healthcare students over an 8-year period. PLoS ONE. 2018;13(3):e0194188. https://doi.org/10.1371/journal.pone.0194188.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Madowitz J, Matheson BE, Liang J. The relationship between eating disorders and sexual trauma. Eat Weight Disord. 2015;20(3):281–93. https://doi.org/10.1007/s40519-015-0195-y.

    Article 
    PubMed 

    Google Scholar 

  • Tavolacci MP, Karmaly A, El Gharbi-Hamza N, Veber B, Ladner J. Gender-based violence among healthcare students: Prevalence, description and associated factors. PLoS ONE. 2023;18(11):e0288855. https://doi.org/10.1371/journal.pone.0288855.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Abaatyo J, Sodeinde SO, Kaggwa MM. Eating disorders among medical students at a private university in Uganda: a cross-sectional study. J Eat Disord. 2025;13(1):56. https://doi.org/10.1186/s40337-025-01234-w.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Bray B, Bray C, Bradley R, Zwickey H. Binge eating disorder is a social justice issue: a cross-sectional mixed-methods study of binge eating disorder experts’ opinions. Int J Environ Res Public Health. 2022;19(10):6243. https://doi.org/10.3390/ijerph19106243.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Wesslund HM, Payne JS, Baxter JD, Westmark DM, Bartels K, Bailey KL, et al. Personal financial wellness curricula for medical trainees: a systematic review. Acad Med. 2023;98(5):636–43. https://doi.org/10.1097/ACM.0000000000005136.

    Article 
    PubMed 

    Google Scholar 

  • Garcia FD, Grigioni S, Allais E, Houy-Durand E, Thibaut F, Déchelotte P. Detection of eating disorders in patients: validity and reliability of the French version of the SCOFF questionnaire. Clin Nutr. 2011;30(2):178–81. https://doi.org/10.1016/j.clnu.2010.09.007.

    Article 
    PubMed 

    Google Scholar 

  • Zigmond AS, Snaith RP. The hospital anxiety and depression scale. Acta Psychiatr Scand. 1983;67(6):361–70. https://doi.org/10.1111/j.1600-0447.1983.tb09716.x.

    Article 
    PubMed 

    Google Scholar 

  • Herrmann C. International experiences with the hospital anxiety and depression scale–a review of validation data and clinical results. J Psychosom Res. 1997;42(1):17–41. https://doi.org/10.1016/s0022-3999(96)00216-4.

    Article 
    PubMed 

    Google Scholar 

  • Gache P, Michaud P, Landry U, Accietto C, Arfaoui S, Wenger O, et al. The alcohol use disorders identification test (AUDIT) as a screening tool for excessive drinking in primary care: reliability and validity of a French version. Alcohol Clin Exp Res. 2005;29(11):2001–7. https://doi.org/10.1097/01.alc.0000187034.58955.64.

    Article 
    PubMed 

    Google Scholar 

  • Tavolacci MP, Grigioni S, Richard L, Meyrignac G, Déchelotte P, Ladner J. Eating Disorders and Associated Health Risks Among University Students. J Nutr Educ Behav. 2015 Sep-Oct;47(5):412–20.e1. https://doi.org/10.1016/j.jneb.2015.06.009.

  • Nagata JM, Ganson KT, Austin SB. Emerging trends in eating disorders among sexual and gender minorities. Curr Opin Psychiatry. 2020;33(6):562–7. https://doi.org/10.1097/YCO.0000000000000645.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Campbell L, Viswanadhan K, Lois B, Dundas M. Emerging evidence: a systematic literature review of disordered eating among transgender and nonbinary youth. J Adolesc Health. 2024;74(1):18–27. https://doi.org/10.1016/j.jadohealth.2023.07.027.

    Article 
    PubMed 

    Google Scholar 

  • Kramer R, Aarnio-Peterson CM, Conard LA, Lenz KR, Matthews A. Eating disorder symptoms among transgender and gender diverse youth. Clin Child Psychol Psychiatry. 2024 Jan;29(1):30–44. https://doi.org/10.1177/13591045231184917.

  • Simone M, Hazzard VM, Askew AJ, Tebbe EA, Lipson SK, Pisetsky EM. Variability in eating disorder risk and diagnosis in transgender and gender diverse college students. Ann Epidemiol. 2022 Jun;70:53–60. https://doi.org/10.1016/j.annepidem.2022.04.007.

  • Simone M, Askew A, Lust K, Eisenberg ME, Pisetsky EM. Disparities in self-reported eating disorders and academic impairment in sexual and gender minority college students relative to their heterosexual and cisgender peers. Int J Eat Disord. 2020 Apr;53(4):513–524. https://doi.org/10.1002/eat.23226.

  • Jahrami H, Sater M, Abdulla A, Faris MA, AlAnsari A. Eating disorders risk among medical students: a global systematic review and meta-analysis. Eat Weight Disord. 2019;24(3):397–410. https://doi.org/10.1007/s40519-018-0516-z.

    Article 
    PubMed 

    Google Scholar 

  • Birmingham WC, Wadsworth LL, Lassetter JH, Graff TC, Lauren E, Hung M. COVID-19 lockdown: impact on college students’ lives. J Am Coll Health. 2023;71(3):879–93. https://doi.org/10.1080/07448481.2021.1909041.

    Article 
    PubMed 

    Google Scholar 

  • Brewerton TD. Eating disorders, trauma, and comorbidity: focus on PTSD. Eat Disord. 2007 Jul-Sep;15(4):285–304. https://doi.org/10.1080/10640260701454311.

  • Rapport de la Mission interministérielle sur les violences sexistes et sexuelles sous relation d’autorité ou de pouvoir, Septembre 2024

  • Association des Etudiants en Médecine de France (ANEMF) Enquête sur les violences sexistes et sexuelles chez les étudiants en médecine 2021 https://anemf.org/iss/enquete-sur-les-vss/

  • Barbanti PCM, Oliveira SRL, de Medeiros AE, Bitencourt MR, Victorino SVZ, Bitencourt MR, et al. Prevalence and impact of academic violence in medical education. Int J Environ Res Public Health. 2022;19(18):11519. https://doi.org/10.3390/ijerph191811519.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Peng P, Hao Y, Liu Y, Chen S, Wang Y, Yang Q, Wang X, Li M, Wang Y, He L, Wang Q, Ma Y, He H, Zhou Y, Wu Q, Liu T. The prevalence and risk factors of mental problems in medical students during COVID-19 pandemic: A systematic review and meta-analysis. J Affect Disord. 2023 Jan 15;321:167–181. https://doi.org/10.1016/j.jad.2022.10.040.

  • Rotenstein LS, Ramos MA, Torre M, Segal JB, Peluso MJ, Guille C, et al. Prevalence of depression, depressive symptoms, and suicidal ideation among medical students: a systematic review and meta-analysis. JAMA. 2016;316(21):2214–36. https://doi.org/10.1001/jama.2016.17324.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Paiva U, Cortese S, Flor M, Moncada-Parra A, Lecumberri A, Eudave L, et al. Prevalence of mental disorder symptoms among university students: an umbrella review. Neurosci Biobehav Rev. 2025;175:106244. https://doi.org/10.1016/j.neubiorev.2025.106244.

    Article 
    PubMed 

    Google Scholar 

  • Römhild A, Hollederer A. Predicting the deviation from the standard study period and dropout intentions through depression severity and social integration among university students in Germany: a longitudinal analysis. Int J Environ Res Public Health. 2025;22(5):667. https://doi.org/10.3390/ijerph22050667.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Mauduy M, Maurage P, Mauny N, Pitel AL, Beaunieux H, Mange J. Predictors of alcohol use disorder risk in young adults: direct and indirect psychological paths through binge drinking. PLoS ONE. 2025;20(5):e0321974. https://doi.org/10.1371/journal.pone.0321974.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Folivi F, Bravo AJ, Pearson MR. Mindfulness Profiles and Substance Use Outcomes in University Students: The Role of Alcohol and Cannabis Use Motives. Mindfulness (N Y). 2025;16(4):1046–1060. https://doi.org/10.1007/s12671-025-02544-5.

  • Tavolacci MP, Déchelotte P, Ladner J. Eating disorders among college students in France: characteristics, help-and care-seeking. Int J Environ Res Public Health. 2020;17(16):5914. https://doi.org/10.3390/ijerph17165914.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Chew-Graham CA, Rogers A, Yassin N. “I wouldn’t want it on my CV or their records”: medical students’ experiences of help-seeking for mental health problems. Med Educ. 2003;37(10):873–80. https://doi.org/10.1046/j.1365-2923.2003.01627.x.

    Article 
    PubMed 

    Google Scholar 

  • Winter RI, Patel R, Norman RI. A Qualitative Exploration of the Help-Seeking Behaviors of Students Who Experience Psychological Distress Around Assessment at Medical School. Acad Psychiatry. 2017 Aug;41(4):477–485. https://doi.org/10.1007/s40596-017-0701-9.

  • Berliant M, Rahman N, Mattice C, Bhatt C, Haykal KA. -Barriers faced by medical students in seeking mental healthcare: A scoping review. MedEdPublish (2016). 2022 Nov 16;12:70. https://doi.org/10.12688/mep.19115.1.

  • Markey CH, August KJ, Rosenbaum DL, Gillen MM, Malik D, Pillarisetty S. An exploratory examination of medical and nursing students’ intentions to discuss body image, weight, and eating disorders with their patients. J Eat Disord. 2024;12(1):159. https://doi.org/10.1186/s40337-024-01119-4.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Continue Reading

  • Cost-Effectiveness Analysis and Priority Setting in the Transition fro

    Cost-Effectiveness Analysis and Priority Setting in the Transition fro

    Introduction

    During pregnancy, adequate maternal nutrition is required to provide the increased nutritional requirements necessary for metabolic maintenance and fetal growth.1–4 In 2020, the Indonesian Ministry of Health documented 4627 maternal deaths, indicating an increase relative to 2019.5 The increase is attributed to the poor nutritional status of pregnant women.6,7 Insufficient multiple micronutrients during pregnancy can increase the risk of complications, including anemia, which affects about 40% of pregnant women globally and peaks at 49% in Southeast Asia.8,9 Multiple micronutrient insufficiency could negatively impact pregnancy outcomes, leading to fetal loss, low birth weight (LBW), preterm birth, preeclampsia, small for gestational age, postpartum depression, elevated risk of neural tube defects, and increased mortality risk.1,10,11 These have been related to immunological development and inadequate neurodevelopmental outcomes in children.12,13 Inadequate development during childhood can extend into adolescence and adulthood, resulting in poor academic performance, reduced income, and reduced human capital.12

    To reduce the risk of various complications, the World Health Organization (WHO) has promoted the supplementation of pregnant women with multiple micronutrient supplementation (MMS) since 2016.14 The administration of MMS is recommended at a dosage of 180 tablets during the first six months of pregnancy.15 MMS is crucial for cellular metabolism, development, and maintaining normal physiological functioning in the human body. MMS is a micronutrient containing 15 vitamins and minerals that fulfill the body’s nutritional requirements. Micronutrient composition as specified by the United Nations International Multiple Micronutrient Antenatal Preparation – multiple micronutrient supplements (UNIMMAP – MMS) consists of vitamin A (800µg), vitamin D (5µg), vitamin E (10mg), vitamin C (70mg), vitamin B1 (1,4mg), vitamin B2 (1,4mg), vitamin B6 (1,6mg), vitamin B12 (2,6 µg), folic acid (400µg), iron (30mg), zinc (15mg), iodine (150µg), selenium (65µg), niacin 18mg, and copper (2mg).16–18 MMS plays essential roles for human reproduction from early pregnancy, facilitating gametogenesis, fertilization, embryogenesis, and the development of placental function, redox balance, and vascularization.2,19–21 MMS contributes a significant role in metabolic processes essential for cell proliferation, growth, and protein synthesis in early pregnancy, and it is crucial for the establishment of the fetal genome throughout gestation. Furthermore, MMS plays an essential role in organogenesis, the development of the fetal central nervous system, and early brain development. MMS is also critical in regulating hemoglobin metabolism and optimizing mitochondrial function during pregnancy.2,11 MMS was shown to reduce oxidative stress and improve mitochondrial function in a study conducted in Lombok, Indonesia.11 As a result, the risk of fetal loss or miscarriage was reduced by 10%, the risk of infant death was reduced by 18%, and the risk of LBW and premature birth was reduced by 14%. This was compared to using iron and folic acid (IFA) alone. Research conducted on a group of pregnant women who suffered from anemia revealed more substantial findings, including a reduction of the risk of fetal loss and neonatal mortality by 29%, a reduction of the risk of newborn death by 38%, and a reduction of the risk of low birth weight by 25%.2

    In October 2024, the Indonesian government started to implement MMS to improve maternal nutrition nationwide, though it remains in the initial phases.22–24 However, the transition from IFA to MMS tablets in Indonesia might take several years and require critical resources, including human capital and financial investment.25 MMS may receive significant government attention and influence healthcare policies. Consequently, the government requires an economic evaluation study as a critical component in the decision-making process. Given Indonesia’s extensive territory, high population density, and the unmet nutritional requirements for pregnant women across its provinces, a comprehensive economic evaluation is necessary to facilitate informed decision-making by policymakers in allocating budgets, resources, and areas of coverage for maternal and child health programs.14,20,26–29 Numerous studies in Bangladesh,14,26–28 India,28,29 Pakistan,28,29 Tanzania,29 Mali,29 and Burkina Faso14 have shown that MMS was a cost-effective intervention compared to IFA in enhancing maternal and child health.14,26–29 This is the first study aimed at evaluating the cost-effectiveness of transitioning from IFA to MMS in Indonesia for enhancing maternal and child health, and its implications for priority setting in Indonesia based on their incremental cost-effectiveness ratio (ICER).

    Methods

    Model Setting

    We applied an open-access online modeling MMS cost-benefit tool developed by Nutrition International to estimate the cost-effectiveness of MMS compared to IFA in Indonesia at the national and sub-national levels (38 provinces). The MMS Tool incorporates a comprehensive set of background data into its foundational model, which has passed quality assurance by technical specialists to guarantee its accuracy, recentness, and relevance.30–32 MMS tools supply national and global policymakers with context-specific assessments that evaluate whether antenatal MMS is more cost-effective compared to IFA.31,32 A 10-year time horizon was applied to assess the cost-effectiveness value, given the substantial effectiveness of MMS in pregnant women. In these scenarios, we applied a coverage scenario at 44% on a national level (baseline), reflecting the current national adherence level of the IFA program in Indonesia, which stands at 44%,33 and a 100% coverage scenario, which is a hypothetical highest cost scenario. The assumption of 180 tablets consumed by each pregnant woman was utilized following WHO recommendations.3,9 The cost-effectiveness analysis of transitioning from IFA to MMS utilized a population of approximately 211,351 pregnant women, as indicated by the 2023 Indonesian Health Survey data.33 We analyzed the projected economic results of MMS compared to IFA at both the national and provincial levels. Additionally, we calculated the ICER by dividing the incremental costs by the disability-adjusted life years (DALYs). The incremental cost-effectiveness ratio (ICER) was calculated using the standardized Cost-Benefit Tool developed by Nutrition International. This tool, which is publicly available on their website (accessible at: https://www.nutritionintl.org/learning-resources-home/mms-cost-benefit-tool/), generates ICER values automatically. Consequently, the results presented in the manuscript are the direct output from this validated tool. The economic value of DALYs averted, reflecting the total economic benefits of transitioning to MMS, is estimated using a monetised DALY approach based on the Value of Statistical Life (VSL). The VSL quantifies the monetary amount an individual is prepared to pay to prevent injury or illness, with variations observed across different countries. Various methods exist for calculating the VSL for a country. This MMS Tool utilizes country-level VSL estimates in LMICs from Viscusi and Masterman34 to derive the Value of a Statistical Life Year (VSLY) by dividing the VSL by the expected life expectancy at birth. The economic value of DALYs averted is calculated by taking the product of the estimated discounted DALYs averted in a particular scenario by the corresponding country’s VSLY. The calculation of the number of DALYs averted incorporates a discount rate of 3%.31,35

    Cost Data

    Costs are determined from the government’s perspective as the health system provider. The MMS cost-benefit modeling tool was designed according to fixed input cost parameters that covered three categories of costs: the cost of IFA, the cost of MMS, and the transition cost (Table 1). These costs are related to a transition from the IFA to the MMS program.30–32 All costs were converted to USD (2024). The cost of MMS was determined using the United Nations International Children’s Emergency Fund (UNICEF) Supply Catalogue.36 The highest price of MMS was set at USD 4 (IDR 61,600) for 180 tablets. The IFA cost was determined by the decree issued by the Indonesian Ministry of Health regarding drug claim prices. Price differences among regions in Indonesia arise from governmental authorities that adjust drug price claims according to logistics expenses, distribution factors, and geographical characteristics specific to each region.37 IFA supplementation requires 180 tablets to ensure 6 months of coverage, and it was priced at USD 3.31 (IDR 50,940).

    Table 1 Breakdown of Costs (2024 USD/ IDR)

    Transition costs include a range of expenditures that extend beyond direct interactions between patients and providers. These include logistical expenses and administrative costs at national and provincial levels, as well as investments in training, nutrition education activities, media promotion, and supervision mechanisms.38,39 In this scenario, the transition cost is set with an additional 13% markup rate applied to the medicine and supply price (ie MMS price) to cover logistics and administrative costs. The costs associated with program transition can constitute an essential component of the total costs.38,39 Table 1 presents the cost data at both the national and provincial levels.

    Maternal Health Parameters

    The effect measure is captured as DALY averted. DALY quantifies the overall disease burden by integrating years of life lost due to premature mortality (YLLs) and years lived with disability (YLDs). One DALY lost signifies one year of healthy life lost; consequently, one DALY averted corresponds to the acquisition of one year of healthy life. Assessing the cost per DALY averted allows for the comparison of various health interventions and the evaluation of an intervention’s impact.26

    The approach examines supplements based on their effectiveness in affecting various health outcomes. This study utilized an open-access online MMS cost-benefit modeling tool developed by Nutrition International,30–32 which was designed according to fixed input maternal health parameters as identified in two published reviews, Keats et al, 201940 and Smith et al, 2017.41 The following maternal health parameters examined in this study, including life expectancy at 73.93 years,33 maternal anemia at 27.7%,33 preterm birth at 11.1%,33 SGA (small for gestational age) at 23.8%, LBW at 6.1%,33 stillbirth at 10.5 per 1000 births,42 maternal mortality at 189 per 100,000 live births,42 neonatal mortality (male and female) at 3.3–3.5 per 1000 live births,42 and infant mortality at 7.8 per 1000 live births.42 These numbers are based on national data from the literature. Sources of health outcomes data include the Indonesian Health Survey 202333 (ie, for preterm birth, maternal anemia, LBW, and life expectancy), the Indonesian Health Profile 202342 (ie, for stillbirth, maternal mortality, neonatal mortality, and infant mortality), and research journals. To determine which provinces should be prioritized in conducting the IFA to MMS transition program, the data collection results were divided into 38 Indonesian provinces (see Table 2).

    Table 2 Maternal Health Parameters

    Priority Setting

    We applied a cost-effectiveness league table to evaluate the prioritization of MMS implementation by comparing the ICER in each province to the national ICER. The cost-effectiveness league table can serve as a tool for resource and budget allocation. A cost-effectiveness league table is a commonly utilized tool for quantitatively ranking priorities based on efficacy, safety, and costs. Healthcare resources could be distributed according to the strategies listed in the league table, beginning with the province with the lowest ICERs and subsequently progressing to the provinces with higher ICERs in the ranking.43 In provincial-level scenarios, we adopted a baseline population coverage of 44%, consistent with the national level.

    Sensitivity Analysis

    We applied one-way deterministic sensitivity analyses to identify variables that might significantly affect results. We performed a one-way deterministic sensitivity analysis to investigate the effects of different input parameters on ICER. All parameters were adjusted by plus or minus 25% for DALYs lost and associated costs.44

    Results

    Maternal Health Outcomes

    Implementing MMS in the 44% and 100% coverage scenarios resulted in 54.897 and 124,766 DALYs averted in Indonesia, respectively. MMS was estimated to have an impact on both scenario (44% and 100% coverage scenarios), resulting in 20,305 and 46,148 DALYs averted in stillbirth, 11,965 and 27,194 DALYs averted in neonatal mortality, 12,879 and 29,271 DALYs averted in preterm birth, 106 and 241 DALYs averted in LBW, 5656 and 12,855 DALYs averted in infant mortality and 9061 and 20,592 DALYs averted in SGA, respectively (Figure S1).

    Cost-Effectiveness Analysis

    According to the perspective of the Indonesian government as the health system provider, the implementation of MMS yielded ICER values of USD 10 per DALY averted in the 44% coverage scenario. Implementing MMS under 100% coverage scenarios yielded an identical ICER value of USD 10 per DALY averted. Variations in coverage will affect the overall costs and benefits, both of which change linearly and proportionally. This is, provided that the ICER, defined as the cost per DALY averted, remains unchanged. The MMS implementation is considered highly cost-effective, as the ICERs remained significantly below the Indonesian one GDP per capita (USD 4870.13) in 2024.45

    Priority Setting

    The MMS implementation in each province is considered highly cost-effective. The ICER of each province in Indonesia is presented in a cost-effectiveness league table (supplementary material 2). Nevertheless, determining priority coverage areas is crucial due to budget constraints and significant disparities in monetary value across different provinces.46 The resource and budget allocation applied to the national ICER as a cost-effectiveness cut-off value. Provinces prioritized for the introduction of MMS in Indonesia are those with ICER values lower than the national ICER value (USD 10 per DALY averted). Therefore, the implementation of the MMS program is recommended to be prioritized in 18 provinces: Southwest Papua, Highlands Papua, Bali, West Java, South Kalimantan, North Maluku, West Papua, Aceh, North Sumatra, Central Java, Central Kalimantan, West Sulawesi, Maluku, Yogyakarta, East Java, North Kalimantan, South Sulawesi, and South Papua as illustrated in Table 3 and Figure 1.

    Table 3 Cost-Effectiveness League Table

    Figure 1 Priority setting for the MMS Program in Indonesia.

    Sensitivity Analysis

    Related to the sensitivity analysis, MMS cost and IFA cost seemed to be the most influential variables affecting the cost-effectiveness value in the implementation of MMS. Other variables that importantly influence the cost-effectiveness value are stillbirth, life expectancy (at birth), neonatal mortality (female), preterm birth, and small gestational age as presented in a tornado chart (Figure S2).

    Discussion

    Insufficient maternal nutritional intake significantly contributes to adverse birth outcomes.1,8,10,11 The WHO has advocated for the supplementation of pregnant women with MMS since 2016 to decrease the risk of numerous complications.9 Our findings show that the projected positive impacts of transitioning from IFA to MMS resulted in reduced mortality rates and adverse birth outcomes. The monetary investment necessary to realize these improvements indicates a cost-effective outcome. Additionally, priority setting based on cost-effectiveness was considered a reasonable option to guide decision-makers in resource allocation to maximize health outcomes, serving as a key consideration in strategic planning for achieving universal health coverage.48,49 Furthermore, the outcomes exhibited stability throughout one-way deterministic sensitivity analyses, indicating that these conclusions were reliable.

    Assuming that all pregnant women adhere to a regimen of 180 pills throughout their pregnancies, the transition from IFA to MMS is projected to 54,897 (44% coverage scenario) and 124,766 (100% coverage scenario) DALYs would be averted in Indonesia. The difference in DALYs averted between the 44% and 100% coverage scenarios is attributable to the differing number of women who benefit from the intervention in each case. With 100% coverage, a greater percentage of pregnant women receive MMS, resulting in a more significant overall decrease in negative maternal and neonatal health outcomes, thus yielding higher DALY estimates. Consuming 180 MMS tablets has played an essential role in pregnant women from early pregnancy in human reproduction, facilitating gametogenesis, fertilization, embryogenesis, placental development, function, redox balance, and vascularization.19–21,50 The growth and function of the placenta are crucial throughout pregnancy to reduce the risk of LBW (14%). MMS demonstrates improved mitochondrial function, which is associated with a reduced risk of premature birth (14%). MMS contributes to hemoglobin metabolism, thus decreasing the risk of anemia in pregnancy. MMS effectively reduces the risk of infant mortality (18%).1,8,10,11,50,51 It is crucial to highlight that, even at existing coverage levels, significant reductions in maternal and child mortality and morbidity are projected in Indonesia if there is high adherence to the prescribed amount of tablets.14 Adherence level is crucial for maximizing the health advantages of MMS intervention20 since medication non-adherence has been identified as a major contributor to health issues and economic burden.52 A report concerning the previous IFA program in Indonesia shows that the adherence rate for the consumption of 90 IFA tablets is merely 44%.33 Consequently, adherence to MMS consumption will be a potential challenge that must be addressed.

    As a country considering investment in MMS, Indonesia requires an assessment of the cost-effectiveness of transitioning from IFA to MMS.29 The MMS cost-benefit tool was utilized to quickly calculate predictions regarding maternal and child health outcomes, as well as the cost-effectiveness of MMS in comparison to IFA for pregnant women. The MMS cost-benefit tool works as an evidence-based modeling instrument designed to facilitate national and international policymakers’ access to data that aids in evaluating the cost-effectiveness of transitioning from IFA to MMS for pregnant women. The MMS cost-benefit tool presents a valuable resource for countries to perform comprehensive, sub-national, and ongoing analyses within the framework of implementation research on MMS.31

    Our findings prove that transitioning from IFA to MMS is highly cost-effective based on the threshold of one to three times Indonesia’s GDP per capita, in the absence of country-specific thresholds.53 Our analysis shows that the implementation of MMS under both 44% and 100% coverage scenarios produced an equal ICER value of USD 10 per DALY averted at the national level. The ICER is consistent across the two scenarios, as it is defined as the additional cost per DALY averted in relation to IFA supplementation. The proportional scaling of both costs and effects with coverage in the modeling framework ensures that the ratio of incremental costs to health benefits remains constant. This demonstrates the difference in the absolute number of DALYs between the two scenarios, despite the constancy of the ICER. The spending required to achieve these improvements, which is below one GDP per capita (USD 4870.13) in 2024, signifies a favorable cost-benefit ratio. Cost-effectiveness analysis provides a quantitative evaluation of both current and prospective efficiency in a health system.49 The cost-effectiveness of the IFA-MMS transition in reducing maternal and child mortality and morbidity (eg, neonatal mortality, infant mortality, LBW, preterm births, stillbirth, and SGA) has proven beneficial. Given that most lives saved would occur in the early stages of life, the policy of transitioning from IFA to MMS is worthy of consideration.

    Currently, MMS is in the initial phases of implementation in Indonesia. The findings of this study support decision-making on the possibility of MMS being expanded as a key focus within Indonesia’s national health program, serving as a primary strategy for lowering the risk of numerous complications in pregnant women.48,49,54 The findings of this study align with previous studies conducted in Bangladesh,14,26–28 Burkina Faso,14 Pakistan,28,29 India,28,29 Mali,29 and Tanzania,29 which indicated that transitioning from IFA to MMS is considered a cost-effective intervention to enhance maternal and child health. This is the first study conducted in Indonesia addressing this issue.

    Related to the sensitivity analyses, the results in this study showed that the cost-effectiveness value was sensitive to changes in MMS cost. Numerous variables can influence MMS costs, including procurement regulations, the volume, and consistency of purchasing bargains with tablet ingredient suppliers, packaging methods, and tablet quantities.14,18,36,55–57 Consequently, the introduction of MMS at accessible costs is essential, particularly for low- and middle-income countries (LMICs), including Indonesia.55 Scale economies in tablet production are crucial therefore, the domestic manufacture of MMS in Indonesia should be initiated to provide a consistent, affordable, and high‐quality supply of MMS, while supporting the expansion of MMS coverage.14,58 Domestic manufacture of MMS could be accomplished by following the standards established by the MMS Technical Advisory Group, alongside a comprehensive technical understanding of the manufacturing prerequisites for the UNIMMAP–MMS product, and the methodologies to ensure that the produced product achieves its expected quality.14,18

    As Islam is the major religion among Indonesia’s population, ensuring halal compliance is essential. The UNIMMAP–MMS product can be produced in compliance with Halal standards established by local authorities.18 Indonesia is home to over 207 million Muslims, representing 87.2% of the population.59 Halal is an essential concept for Muslim consumers concerning the products they consume, including pharmaceutical ingredients. In contemporary medicine, these ingredients must be completely free of porcine (0%) and contain less than 1% alcohol.60,61

    In October 2024, the Indonesian Ministry of Health announced the immediate initiation of MMS in Indonesia, a program currently in its early stages.22–24 Switching from IFA to MMS tablets in Indonesia may require several years and substantial resources, particularly personnel and budgetary expenditure, given the country’s expansive territory and dense population.25 Due to logistical and budget limitations, determining priority coverage areas is essential to mitigate the disease burden associated with nutritional deficiencies among pregnant women. This involves identifying higher-risk groups and regions where preventive measures are expected to be most effective, thereby maximizing public health returns on investment.20,46

    Priority settings should focus on optimizing population health, and the availability of further knowledge regarding cost-effectiveness will enhance decision-making and result in improved health outcomes as one consideration in strategic planning.48,49 Cost-effectiveness evidence in each province enables policy-makers to assess the effective and efficient use of available resources. It also guides optimal investment strategies to meet health targets and achieve universal health coverage within the constraints of limited resources, ensuring the optimal allocation of financial resources in the healthcare sector.38,49 Universal health coverage signifies that every individual has access to a comprehensive array of quality health services as required, without experiencing financial difficulties.62 Based on the results of the cost-effectiveness analysis, it’s recommended to prioritize introducing the MMS program in 18 provinces where the ICER is below Indonesia’s national ICER of USD 10. The analysis commences with the province exhibiting the lowest ICERs, subsequently advancing to those with higher ICERs in the rank order.43 Public health strategies prioritize introducing MMS programs, especially in provinces with poor maternal health outcomes. Expanding MMS programs in LMICs necessitates advancements in supply chain logistics and improved availability and access to health services.15,63 The prioritization of MMS implementation allows for the distribution of healthcare resources under the strategies, starting with the province exhibiting the lowest cost-effectiveness and subsequently advancing to those with higher cost-effectiveness in the ranking.

    To the best of our knowledge, our study is the first to assess cost-effectiveness analysis regarding the transition from IFA to MMS during pregnancy in Indonesia at the national and sub-national levels (38 provinces). The primary strength of this study is the use of country-specific data, which enables policymakers to make informed decisions regarding finance and resource allocation by prioritizing selected coverage areas in MMS implementation. Specifically, MMS programs have not yet been included in the national healthcare insurance coverage unit, and our study offers valuable insights into the implications of their potential inclusion. With regards to the shifting from IFA to MMS supplementation in Indonesia, our cost-effectiveness study demonstrates that the implementation of MMS is more advantageous than IFA, aligning with the WHO’s recommendation to improve the quality of maternal and child health.

    Nevertheless, this study has several limitations. The first limitation concerns a proposed transition cost that has been estimated based on population size to assess context-specific expenses associated with initiating a new program. These expenses include developing training components, establishing new policies and regulations, and training healthcare personnel; however, actual costs may differ. In the absence of more reliable data on transition costs, we perform a costing exercise for transition activities informed by a published article on the cost-effectiveness of interventions aimed at improving maternal, newborn, and child health outcomes: a WHO Choosing Interventions that are Cost-Effective (CHOICE) analysis for Eastern Sub-Saharan Africa and South-East Asia.38 Consequently, we may have underestimated the transition cost. As for the second limitation, this study did not address MMS adherence. The analysis presumes that all “covered” pregnant women receive and take precisely 180 tablets. Achieving all consumption of a precise dosage of tablets by all covered women will be challenging and does not seem to occur systematically. Some pregnant women may take fewer than 180 tablets. Referring to the previous IFA program in Indonesia, a study indicates that the adherence level to the consumption of 90 IFA tablets is only 44%.33 The costs associated with transitioning from IFA to MMS will rise, while the expected benefits will remain unchanged. Further attempts are required to address these issues. The third limitation is that the modeling tool for cost-benefit analysis, developed by Nutrition International, does not allow the inclusion of additional maternal health parameters that could be significant for assessing the effectiveness of MMS in pregnant women. The model focuses on health outcomes of interest, as identified in two published reviews, Keats et al, 201940 and Smith et al, 2017.41 The fourth limitation is that probabilistic sensitivity analysis (PSA) cannot be performed using the open-access online MMS cost-benefit tool developed by Nutrition International because the tool is designed primarily to provide deterministic estimates based on fixed input parameters rather than full probabilistic modeling. While the tool incorporates rigorous methodologies to estimate health impacts, costs, and cost-effectiveness, it does not support Monte Carlo simulations or the input of probability distributions for parameters that allow quantification of uncertainty across multiple model inputs simultaneously. However, the deterministic approach can be used to provide clear, interpretable results such as benefit-cost ratios and incremental cost per DALY averted based on fixed assumptions, but without performing full probabilistic uncertainty analysis. More complex PSA requiring simulation of parameter distributions and joint uncertainty are typically conducted offline using statistical software, as seen in some detailed cost-effectiveness studies of MMS interventions. Thus, while the tool does conduct some sensitivity analyses by varying key assumptions, full probabilistic sensitivity analysis is not supported in its open-access online version.

    This study provides recommendations for policymakers in Indonesia to decide on following comprehensive policies to improve maternal and child health outcomes. Health enhancements throughout gestation and early childhood might lower the likelihood of poor health and disease, hence supporting life expectancy.64 The additional budgetary requirement poses significant challenges to implement in a country with constrained healthcare budgets for maternal and child health programs. The MMS implementation strategy could start with a 44% coverage scenario across 18 prioritized provinces and gradually expand to 100% coverage for all pregnant women in Indonesia. This approach is critical for transitioning from the IFA to the MMS program. Transitioning from IFA to MMS represents the ideal approach for enhancing maternal and child health outcomes.54 We are encouraged that the reviewer acknowledges the policy relevance of our findings and their potential to inform resource allocation and program planning for maternal nutrition interventions. Further study is required to incorporate a concise perspective on the necessity of supplementary qualitative or operational studies, which could highlight the significance of converting economic findings into practical implementation.

    Conclusions

    This study showed the ICER value of USD 10 per DALY averted for both scenarios (44% and 100% coverage), concluding that the transition from IFA to the MMS program was confirmed to be a highly cost-effective intervention. These results should facilitate decision-making that prioritizes maternal and child health. The MMS implementation strategy could start with a 44% coverage scenario across 18 prioritized provinces and gradually expand to 100% coverage for all pregnant women in Indonesia. This strategy is essential for the transition from the IFA to the MMS program in Indonesia. It is essential to identify several potential barriers to the implementation of the MMS program in Indonesia, including the adequacy of supply chain logistics, the improvement of health service availability and access, and the need for better adherence to MMS consumption.

    Ethical Clearance

    Data were obtained from publicly accessible documents, and human participants were not involved in this investigation. Consequently, ethical considerations regarding human participants were not necessary. Nevertheless, efforts were made to guarantee that the data were collected and analyzed in a manner that was both ethical and transparent.

    Disclosure

    The authors report no conflicts of interest in this work.

    References

    1. Massari M, Novielli C, Mandò C, et al. Multiple micronutrients and docosahexaenoic acid supplementation during pregnancy: a randomized controlled study. Nutrients. 2020;12(8):1–16. doi:10.3390/nu12082432

    2. Bourassa MW, Osendarp SJM, Adu-Afarwuah S, et al. Review of the evidence regarding the use of antenatal multiple micronutrient supplementation in low- and middle-income countries. Annals of the New York Academy of Sciences. Vol. 1444. The New York Academy of Sciences, New York, NY, United States:Blackwell Publishing Inc.;2019:6–21. Available from: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85066409444&doi=10.1111%2Fnyas.14121&partnerID=40&md5=50806e658b69069ee0e9fe036cffc927.

    3. Tuncalp Ö, Rogers LM, Lawrie TA, et al. WHO recommendations on antenatal nutrition: an update on multiple micronutrient supplements. BMJ Glob Heal. 2020;5(7):e003375. doi:10.1136/bmjgh-2020-003375

    4. Gomes F, Askari S, Black RE, et al. Antenatal multiple micronutrient supplements versus iron-folic acid supplements and birth outcomes: analysis by gestational age assessment method. Matern Child Nutr. 2023;19(3). doi:10.1111/mcn.13509

    5. Kesehatan RIK. Profil kesehatan Indonesia tahun 2020. Kementerian Kesehatan republik Indonesia: 2021.

    6. Manfredini M. The effects of nutrition on maternal mortality: evidence from 19th-20th century Italy. SSM – Popul Heal. 2020;12:100678. doi:10.1016/j.ssmph.2020.100678

    7. Bakshi RK, Kumar N, Srivastava A, et al. Decadal trends of maternal mortality and utilization of maternal health care services in India: evidence from nationally representative data. J Fam Med Prim Care. 2025;14(5):1807. doi:10.4103/jfmpc.jfmpc_916_24

    8. Berti C, Gaffey MF, Bhutta ZA, Cetin I. Multiple-micronutrient supplementation: evidence from large-scale prenatal programmes on coverage, compliance and impact. Matern Child Nutr. 2018;14 Suppl 5(Suppl 5):e12531. doi:10.1111/mcn.12531

    9. World Health Organization. WHO antenatal care recommendations for a positive pregnancy experience. nutritional intervention update: multiple micronutrient supplements during pregnancy. World Health Organization: 2020.

    10. Petry CJ, Ong KK, Hughes IA, Dunger DB. Multiple micronutrient supplementation during pregnancy and increased birth weight and skinfold thicknesses in the offspring: the Cambridge baby growth study. Nutrients. 2020;12(11):1–13. doi:10.3390/nu12113466

    11. Priliani L, Prado EL, Restuadi R, Waturangi DE, Shankar AH, Malik SG. Maternal multiple micronutrient supplementation stabilizes mitochondrial DNA copy number in pregnant women in Lombok, Indonesia. J Nutr. 2019;149(8):1309–1316. doi:10.1093/jn/nxz064

    12. Sudfeld CR, Bliznashka L, Salifou A, et al. Evaluation of multiple micronutrient supplementation and medium-quantity lipidbased nutrient supplementation in pregnancy on child development in rural Niger: a secondary analysis of a cluster randomized controlled trial. PLoS Med. 2022;19(5):1–17. doi:10.1371/journal.pmed.1003984

    13. Moore SE, Fulford AJ, Darboe MK, Jobarteh ML, Jarjou LM, Prentice AM. A randomized trial to investigate the effects of pre-natal and infant nutritional supplementation on infant immune development in rural Gambia: the ENID trial: early nutrition and immune development. BMC Pregnancy Childbirth. 2012;12(1):107. doi:10.1186/1471-2393-12-107

    14. Engle-Stone R, Kumordzie SM, Meinzen-Dick L, Vosti SA. Replacing iron-folic acid with multiple micronutrient supplements among pregnant women in Bangladesh and Burkina Faso: costs, impacts, and cost-effectiveness. Annals of the New York Academy of Sciences. Vol. 1444. Department of Nutrition, University of California – Davis, Davis, CA, United States:Blackwell Publishing Inc.;2019:35–51. Available from: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85065195150&doi=10.1111%2Fnyas.14132&partnerID=40&md5=01f9ee9fec236784fb85abdc3703bf3c.

    15. MMS TAG. Focusing on multiple micronutrient supplements in pregnancy: second edition. update on the scientific evidence on the benefits of prenatal multiple micronutrient supplements. 2023: 15–22. Available from: https://d2b2stjpsnac9i.cloudfront.net/wp-content/uploads/2023/05/202305-MMS-2-sightandlife.pdf#page=15. Accessed November 04, 2025.

    16. Haider BA, Bhutta ZA. Multiple-micronutrient supplementation for women during pregnancy. Cochrane Database Syst Rev. 2012;11(11):CD004905. doi:10.1002/14651858.CD004905.pub3

    17. Schaefer E, Nock D. The impact of preconceptional multiple-micronutrient supplementation on female fertility. Clin Med Insights Women’s Heal. 2019;12:1179562X1984386.

    18. MMS-TAG, MNF. Expert consensus on an open-access united nations international multiple micronutrient antenatal preparation-multiple micronutrient supplement product specification. Ann N Y Acad Sci. 2020;1470(1):3–13. doi:10.1111/nyas.14322

    19. Mei Z, Jefferds ME, Namaste S, Suchdev PS, Flores-Ayala RC. Monitoring and surveillance for multiple micronutrient supplements in pregnancy. Matern Child Nutr. 2018;14(April 2017):1–9. doi:10.1111/mcn.12501

    20. Gomes F, Bourassa MW, Adu-Afarwuah S, et al. Setting research priorities on multiple micronutrient supplementation in pregnancy. Ann N Y Acad Sci. 2020;1465(1):76–88. doi:10.1111/nyas.14267

    21. Schulze KJ, Gernand AD, Khan AZ, et al. Newborn micronutrient status biomarkers in a cluster-randomized trial of antenatal multiple micronutrient compared with iron folic acid supplementation in rural Bangladesh. Am J Clin Nutr. 2020;112(5):1328–1337. doi:10.1093/ajcn/nqaa223

    22. Kemenkes R. Keputusan menteri kesehatan republik indonesia nomor HK.01.07/MENKES/1092/2024 tentang standar suplemen zat gizi mikro untuk ibu hamil. 2024: 1–8.

    23. Vitamin Angels. How Indonesia is transforming maternal nutrition with MMS | vitamin angels. 2024. Available from: https://vitaminangels.org/news/how-indonesia-is-transforming-maternal-nutrition-with-mms/. Accessed March 5, 2025.

    24. Menkes tekankan pentingnya ragam mikronutrien bagi ibu hamil. Available from: https://kemkes.go.id/id/rilis-kesehatan/menkes-tekankan-pentingnya-ragam-mikronutrien-bagi-ibu-hamil. Accessed March 7, 2025.

    25. Meinzen-dick L, Vosti SA. The evidence base cost-effectiveness of replacing iron-folic acid with multiple micronutrient supplements.

    26. Svefors P, Selling KE, Shaheen R, Khan AI, Persson L-Å, Lindholm L. Cost-effectiveness of prenatal food and micronutrient interventions on under-five mortality and stunting: analysis of data from the MINIMat randomized trial. Bangladesh PLoS One. 2018;13(2):e0191260.

    27. Shaheen R, Persson LÅ, Ahmed S, Streatfield PK, Lindholm L. Cost-effectiveness of invitation to food supplementation early in pregnancy combined with multiple micronutrients on infant survival: analysis of data from MINIMat randomized trial, Bangladesh. BMC Pregnancy Childbirth. 2015;15(1):125. doi:10.1186/s12884-015-0551-y

    28. Kashi B, Godin CM, Kurzawa ZA, Verney AMJ, Busch-Hallen JF, De-Regil LM. Multiple micronutrient supplements are more cost-effective than iron and folic acid: modeling results from 3 high-burden Asian countries. J Nutr. 2019;149(7):1222–1229. doi:10.1093/jn/nxz052

    29. Young N, Bowman A, Swedin K, et al. Cost-effectiveness of antenatal multiple micronutrients and balanced energy protein supplementation compared to iron and folic acid supplementation in India, Pakistan, Mali, and Tanzania: a dynamic microsimulation study. PLoS Med. 2022;19(2):e1003902. doi:10.1371/journal.pmed.1003902

    30. MMS cost-benefit tool – nutrition international. Available from: https://www.nutritionintl.org/learning-resources-home/mms-cost-benefit-tool/. Accessed March 17, 2025.

    31. Verney AMJ, Busch-Hallen JF, Walters DD, Rowe SN, Kurzawa ZA, Arabi M. Multiple micronutrient supplementation cost-benefit tool for informing maternal nutrition policy and investment decisions. Matern Child Nutr. 2023;19(4):e13523. doi:10.1111/mcn.13523

    32. Interface U, Guide I. The MMS Cost-Benefit Tool. 2019.

    33. Kemenkes R. Survei Kesehatan Indonesia (SKI) 2023. Kemenkes RI; 2023.

    34. Viscusi WK, Masterman CJ. Income elasticities and global values of a statistical life. J Benefit-Cost Anal. 2017;8(2):226–250. doi:10.1017/bca.2017.12

    35. Robinson LA, Hammitt JK, Cecchini M, et al. Reference case guidelines for benefit-cost analysis in global health and development. SSRN Electron J. 2022;(May).

    36. UNICEF. Multiple Micronutrient Powder Supply and Market Update. Copenhagen, Denmark: UNICEF Supply Division; 2021.

    37. Kemenkes R. Nilai klaim harga obat program rujuk balik, obat penyakit kronis di fasilitas kesehatan rujukan tingkat lanjutan, obat kemoterapi, dan obat alteplase. keputusan menteri kesehat republik indones no HK0107/MENKES/1905/2023. 2023: 1–58.

    38. Stenberg K, Watts R, Bertram MY, et al. Cost-effectiveness of interventions to improve maternal, newborn and child health outcomes: a WHO-CHOICE analysis for Eastern Sub-Saharan Africa and South-East Asia. Int J Heal Policy Manag. 2021;10(11):706–723.

    39. Johns B, Baltussen R, Hutubessy R. Programme costs in the economic evaluation of health interventions. Cost Eff Resour Alloc. 2003;1(1):1–10. doi:10.1186/1478-7547-1-1

    40. Keats EC, Haider BA, Tam E, Bhutta ZA. Multiple-micronutrient supplementation for women during pregnancy (Review). Cochrane Database Syst Rev. 2019;2019(3).

    41. Smith ER, Shankar AH, Wu LS, et al. Modifiers of the effect of maternal multiple micronutrient supplementation on stillbirth, birth outcomes, and infant mortality: a meta-analysis of individual patient data from 17 randomised trials in low-income and middle-income countries. Lancet Glob Heal. 2017;5(11):e1090–100. doi:10.1016/S2214-109X(17)30371-6

    42. Kementrian Kesehatan. Profil kesehatan Indonesia 2023. 2024: 550.

    43. Mauskopf J, Rutten F, Schonfeld W. Cost-effectiveness league tables. Valuable guidance for decision makers? PharmacoEconomics – Ital Res Artic. 2004;6(3):131–140. doi:10.1007/BF03320631

    44. Zakiyah N, van Asselt ADI, Setiawan D, Cao Q, Roijmans F, Postma MJ. Cost-effectiveness of scaling up modern family planning interventions in low- and middle-income countries: an economic modeling analysis in Indonesia and Uganda. Appl Health Econ Health Policy. 2019;17(1):65–76. doi:10.1007/s40258-018-0430-6

    45. Indonesia B. Statistik Indonesia Statistical Yearbook of Indonesia 2024. Vol. 1101001. BPS-Statistics Indonesia; 2024:790.

    46. Mori AT, Robberstad B. Pharmacoeconomics and its implication on priority-setting for essential medicines in Tanzania: a systematic review. BMC Med Inform Decis Mak. 2012;12(1):1. doi:10.1186/1472-6947-12-110

    47. Produk domestik regional bruto per kapita atas dasar harga berlaku menurut provinsi (ribu rupiah), 2024 – tabel statistik – badan pusat statistik Indonesia. Available from: https://www.bps.go.id/id/statistics-table/3/YWtoQlRVZzNiMU5qU1VOSlRFeFZiRTR4VDJOTVVUMDkjMw==/produk-domestik-regional-bruto-per-kapita-atas-dasar-harga-berlaku-menurut-provinsi–ribu-rupiah-2022.html?year=2023. Accessed June 18, 2025.

    48. Baltussen R, Jansen MP, Mikkelsen E, et al. Priority setting for universal health coverage: we need evidence-informed deliberative processes, not just more evidence on cost-effectiveness. Int J Heal Policy Manag. 2016;5(11):615–618. doi:10.15171/ijhpm.2016.83

    49. Bertram MY, Lauer JA, Stenberg K, Edejer TTT. Methods for the economic evaluation of health care interventions for priority setting in the health system: an update from WHO CHOICE. Int J Heal Policy Manag. 2021;10(11):673–677.

    50. Bourassa MW, Osendarp SJM, Adu-Afarwuah S, et al. Review of the evidence regarding the use of antenatal multiple micronutrient supplementation in low- and middle-income countries. Ann N Y Acad Sci. 2019;1444(1):6–21. doi:10.1111/nyas.14121

    51. Haider BA, Bhutta ZA. Multiple-micronutrient supplementation for women during pregnancy. Cochrane Database Syst Rev. 2017;4(4):CD004905. doi:10.1002/14651858.CD004905.pub5

    52. Octaviani P, Ikawati Z, Yasin NM, Kristina SA, Kusuma IY. Interventions to improve adherence to medication on multidrug-resistant tuberculosis patients: a scoping review. Med J Malaysia. 2024;79(2):212–221.

    53. Marseille E, Larson B, Kazi DS, Kahn JG, Rosen S. Thresholds for the cost–effectiveness of interventions: alternative approaches. Bull World Health Organ. 2015;93(2):118–124. doi:10.2471/BLT.14.138206

    54. Alfiani F, Meita Utami A, Zakiyah N, Aizati Athirah Daud N, Suwantika AA, Puspitasari IM. Cost-effectiveness analysis of multiple micronutrient supplementation (MMS) compared to iron folic acid (IFA) in pregnancy: a systematic review. Int J Womens Health. 2025;17:639–649. doi:10.2147/IJWH.S489159

    55. Zakiyah N, Insani WN, Suwantika AA, van der Schans J, Postma MJ. Pneumococcal vaccination for children in Asian countries: a systematic review of economic evaluation studies. Vaccines. 2020;8(3):1–18. doi:10.3390/vaccines8030426

    56. Paulden M, O’Mahony J, McCabe C. Determinants of Change in the Cost-effectiveness Threshold. Med Decis Mak. 2017;37(2):264–276. doi:10.1177/0272989X16662242

    57. Garcia-Casal MN, Estevez D, De-Regil LM. Multiple micronutrient supplements in pregnancy: implementation considerations for integration as part of quality services in routine antenatal care. Objectives, results, and conclusions of the meeting. Matern Child Nutr. 2018;14(S5). doi:10.1111/mcn.12704

    58. Monterrosa EC, Beesabathuni K, van Zutphen KG, et al. Situation analysis of procurement and production of multiple micronutrient supplements in 12 lower and upper middle-income countries. Matern Child Nutr. 2018;14(S5). doi:10.1111/mcn.12500

    59. Laman resmi republik Indonesia • Portal informasi Indonesia. Available from: https://indonesia.go.id/profil/agama. Accessed March 15, 2025.

    60. Herdiana Y, Sofian FF, Shamsuddin S, Rusdiana T. Towards halal pharmaceutical: exploring alternatives to animal-based ingredients. Heliyon. 2023;10(1). doi:10.1016/j.heliyon.2023.e23624

    61. Pemerintah Indonesia. Peraturan presiden (PERPRES) nomor 6 tahun 2023 tentang sertifikasi halal obat, produk biologi, dan alat kesehatan. Perpres Nomor. 2023;(148729):1–17.

    62. World Health Organization. Universal health coverage (UHC). 2023. Available from: https://www.who.int/news-room/fact-sheets/detail/universal-health-coverage-uhc. Accessed March 13, 2025.

    63. Shekar M, Kakietek J, Dayton EJ, Walters D. An investment framework for nutrition. 2016.

    64. Dundas R, Boroujerdi M, Browne S, et al. Evaluation of the Healthy Start voucher scheme on maternal vitamin use and child breastfeeding: a natural experiment using data linkage. Public Heal Res. 2023;11(11):1–101.

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  • Current State and Challenges of Dental Intervention in Aspiration Pneu

    Current State and Challenges of Dental Intervention in Aspiration Pneu

    Introduction

    The world is trending toward an aging society, and this trend is expected to continue.1 In Japan, this impact is particularly pronounced, with projections suggesting that by 2060, nearly 40% of the population will be older than 65 years.2 In addition, the incidence of aspiration pneumonia in Japan increased by 53.7% between 2005 and 2019.3 The United States has a similar substantial burden: a recent epidemiological analysis identified over 1.1 million aspiration pneumonia-related deaths from 1999 to 2017, averaging about 58,000 deaths per year.4 Although the incidence decreased between 2002 and 2012, the median total cost of hospitalization for patients aged ≥ 65 years doubled over the decade, rising from US $16,173 to US $30,280.5 Addressing these issues has become an urgent global challenge.4,6

    Generally, the risk of aspiration pneumonia is influenced by diminished swallowing function and the intraoral environment.7 In particular, disruption of the oral microbiome and biofilm formation is a core mechanism linking oral conditions to systemic health.8 Poor oral hygiene plays a pivotal role in this process by increasing the oropharyngeal bacterial load. Pathogenic microorganisms that proliferate in unclean mouths, including anaerobes, such as Fusobacterium spp., and Gram-negative bacilli, such as Klebsiella pneumoniae, can be aspirated into the lungs, especially in patients with dysphagia or an altered gag reflex.9 Therefore, it has been shown that indicators of good oral hygiene, such as receiving regular professional dental cleanings and frequent toothbrushing (≥3 times a day), are independently associated with lower pneumonia incidence.10 Also, specialized oral health care provided by dentists and dental hygienists has been shown to significantly reduce the incidence of aspiration pneumonia during the perioperative period for esophageal cancer.11 Furthermore, recent reports suggest that oral health care by dentists can prevent the recurrence of aspiration pneumonia, highlighting the critical importance of dental intervention in the management of aspiration pneumonia.12–14

    Given the clear link between oral health interventions and pneumonia, dental professionals have a vital role to play in the multidisciplinary management of aspiration pneumonia. However, some barriers persist in achieving collaboration between physicians and dentists. One challenge is the separation of the healthcare system: in many countries, dentistry has operated largely in parallel to medicine rather than integrated within it.15 Also, the number of dentists in hospitals is limited: only 28% of hospitals in Japan currently employ full-time dentists.16 Furthermore, another barrier is the limited awareness and training across professions. Some physicians and nurses underestimate the impact of oral hygiene on systemic health or view oral care as a “nursing task” rather than a medical priority.14,17

    Hospitalists, through their interventions, have been reported to reduce the length of hospital stays, improve the quality of medical care, and have a favorable impact on healthcare economics.18,19 Also, it is shown that hospitalist management of pneumonia is associated with shorter length of stay and lower costs compared with non-hospitalist care.20 Although they are familiar with aspiration pneumonia, to the best of our knowledge, there have been no investigations into the frequency or specific nature of hospitalists’ collaboration with dentists in the management of aspiration pneumonia.

    Therefore, we aimed to clarify the state of dental involvement in aspiration pneumonia management and identify challenges that need to be addressed.

    Materials and Methods

    Study Design, Setting, and Participants

    This study was an observational cross-sectional survey based on questionnaires sent to all individuals listed on the Japanese Society of Hospital General Medicine (JSHGM) mailing lists. JSHGM is primarily responsible for the board certification of hospitalists, whereas the Japan Primary Care Association (JPCA) is mainly responsible for the board certification of family physicians. Many hospitals in Japan belong to both organizations. Since this study focused on hospitalists, the survey was conducted among members of JSHGM,21 as it was considered suitable for surveying hospitalists in Japan. In this study, hospitalists were defined as general medicine doctors working in a hospital, which was defined as a hospital with 20 or more beds.22 The participants of this study were Japanese hospitalists. We included data collected from August 23 to November 15, 2023. Individuals who did not provide consent and those who were not hospitalists were excluded.

    Survey Instrument Variable

    In this study, we developed an original questionnaire to collect baseline data on participants’ basic attributes and work environments, as well as outcome data on the extent to which dentistry-related practices are being implemented (Appendix Figure 1). Baseline data included age, gender, years of experience as a doctor, type of hospital (community-based hospital or university hospital), the presence or absence of an oral surgery department, the number of full-time dentists (0, 1, 2, or 3 or more), and the availability of dental hygienists.

    For the outcome data, we examined whether the following actions, considered important steps in consulting an oral surgery department when treating aspiration pneumonia, were performed: oral evaluation, use of oral healthcare assessment tools, checking dentures, identifying the patient’s primary care dentist, recommending a dental visit after discharge, and encouraging a dental visit for patients with relevant medical histories. These items were evaluated in 20% increments (0%, 1–20%, 21–40%, 41–60%, 61–80%, 81–100%) and recategorized into four groups: 0% as “never”, 1–40% as “sometimes”, 41–80% as “often”, and 81–100% as “always”.

    Our primary outcome was whether respondents made any dental referral for aspiration-pneumonia management (0% vs 1–100%). A prespecified secondary outcome was routine referral, defined as a referral frequency of 81–100%. Explanatory variables comprised (i) system-level resources—the presence of an oral surgery department, the number of full-time dentists, the presence of dental hygienists, and hospital type; (ii) clinician practices—six oral-care activities (oral evaluation, use of an oral health assessment tool, denture check, identifying the patient’s primary-care dentist, recommending a dental visit after discharge, and encouraging a dental visit in those with a relevant history), each recorded on a six-category frequency scale and recoded as never/sometimes/often/always; and (iii) demographics—age, sex, and years in practice.

    We evaluated whether respondents initiated dental referrals for the management of aspiration pneumonia by classifying their referral frequency into 20% increments (0%, 1–20%, 21–40%, 41–60%, 61–80%, 81–100%). Those who referred between 1% and 100% were assigned to the “refer group”, while those with 0% referral were categorized as the “non-refer group”. Within the Refer group, individuals referring 81–100% of the time were identified as the “routinely refer group”. Furthermore, the referral rate was examined among respondents who reported performing multiple actions in the always group. Specifically, we identified respondents who consistently performed both “oral evaluation” and “use of oral healthcare assessment tools” or both “oral evaluation” and “checking dentures” in the always group. Among these respondents, the routinely refer group was calculated and compared with the proportion in the non-refer group.

    Respondents were also asked, through multiple-choice questions, to indicate their reasons for referring or not referring. These options were determined through discussions involving TM, ST, and TM (Matsumoto). We then analyzed the clinical practices (“always”) routinely performed by hospitalists in the Refer group. Furthermore, we analyzed the sum of these practices.

    Data Analysis

    All statistical analyses were performed using JMP version 18.1 (SAS Institute, Cary, NC). Patient-level variables are presented as medians and interquartile ranges (IQR) for continuous variables and as numbers and percentages for categorical variables. For continuous variables, the Mann–Whitney U-test was used after normality was assessed. Statistical significance was defined as a p-value of < 0.05 using a two-tailed test. Regarding missing data, analyses were conducted using only the non-missing portions of the dataset.

    Ethics

    The study was approved by the ethics committee of the Ashikaga Red Cross Hospital (No.2024–34) and was conducted in accordance with the principles of the Declaration of Helsinki. All participants reviewed the study document detailing data anonymization, voluntary participation, and the dissemination of research results prior to participation. Only participants who provided informed consent (opt-in) were included in the study. Additionally, participants could withdraw from the study at any time.

    Results

    A total of 370 hospitalists participated in this study. Of these, 18 worked in clinics and were excluded from the analysis, leaving 352 participants in the final study sample.

    Among the participants, 305 (86.7%) were male, and the median age (IQR) was 48 (40–56) years. The median number of years of practice was 22 (13–31) years. A total of 255 (72.4%) worked at community-based hospitals, and 234 (66.4%) worked at institutions with an affiliated oral surgery department. Regarding the number of full-time dentists, hospitals with three or more full-time dentists were common, accounting for 159 participants (45.1%). Finally, 237 participants (67.3%) reported having dental hygienists available at their workplaces.

    Among these participants, 141 (40.1%) referred patients for dental consultation as part of their treatment for aspiration pneumonia. In this group, the following factors showed significant differences. Hospitalists whose hospitals had an affiliated oral surgery department were more likely to make referrals (affiliated: 118 [50.4%] vs unaffiliated: 23 [19.5%], P<0.001). Similarly, those working in hospitals with full-time dentists showed a higher referral rate (0 full-time dentists: 26 [20.8%], 1 full-time dentist: 12 [42.9%], 2 full-time dentists: 25 [62.5%], 3 or more full-time dentists: 78 [49.1%], P<0.001). A similar difference was observed regarding the presence of dental hygienists (presence: 118 [49.8%] vs absence: 23 [20%]; P<0.001). Moreover, hospitalists at university hospitals were more likely to make referrals than those at community-based hospitals (university hospitals: 51 [52.6%] vs community-based hospitals: 90 [35.3%]; P<0.01) (Table 1).

    Table 1 Participant Characteristics

    Furthermore, among the participants, the proportion of those who routinely performed the following practices in aspiration pneumonia care (always group) was as follows: 138 (39.2%) reported conducting oral evaluations themselves for patients admitted with aspiration pneumonia. Of these, 37 (11.3%) used the oral health assessment tool. In addition, 169 (48.0%) checked whether patients had dentures. Only 30 (8.5%) recommended a dental visit after discharge, and 36 (10.2%) encouraged dental visits for patients with a history of aspiration pneumonia (Table 2).

    Table 2 Regarding the Treatment of Aspiration Pneumonia by Japanese Hospitalists

    A total of 126 (89.4%) participants were referred for oral healthcare, 85 (60.3%) for denture adjustment, 61 (43.3%) for swallowing function assessment, and 58 (41.1%) for tooth extraction. Meanwhile, 95 (45.0%) did not make referrals because they consulted other healthcare professionals, such as speech-language therapists and nurses. Additionally, 91 (43.1%) cited a lack of a habit of making referrals and 87 (41.2%) mentioned the absence of a dentist in the hospital as reasons for not referring patients (Table 3).

    Table 3 The Reasons for Each Group of Patients Who Were (a) Referred to a Dentist or (b) Not Referred to a Dentist

    The referral frequency results for those who routinely referred patients were as follows: among those in the always oral evaluation group, 75 (54.3%) referred patients with aspiration pneumonia. Within this group, 25 (69.4%) who routinely checked for a primary care dentist had a higher referral rate. Similarly, in the always group, 22 (59.5%) who used the oral healthcare assessment tool referred patients when they developed aspiration pneumonia. Moreover, 14 (73.7%) in this group who routinely consulted their primary care dentists also had a high referral rate (Table 4).

    Table 4 Comparison of Medical Care Between Groups That are Always and are Not Introduced

    Discussion

    This study investigated the involvement of dentists in the management of aspiration pneumonia among hospitalists in Japan. Our findings revealed that the most common reason for not referring patients was consultation with other non-dental professionals. This study identified the hospital and practice characteristics of hospitalists who routinely referred patients to dentists.

    Reasons Why Hospitalists Do Not Refer Patients to Dentists

    In this study, the main reasons for not referring patients to dentists were consultation with other medical professionals, such as speech-language therapists and nurses, lack of a referral habit, and absence of dentists within the hospital. The first reason was reliance on others. Oral bacteria predispose patients to aspiration pneumonia, and previous studies have suggested that oral health care provided by dentists can reduce bacterial levels.23,24 Although oral rehabilitation by speech-language therapists has been shown to shorten hospital stay and improve outcomes,25 they are unable to provide oral health care themselves, making it difficult to fully address oral health issues. In contrast, while 80.2% of nurses recognized the importance of oral health care in preventing aspiration pneumonia, studies suggest that collaborative care with dentists is more effective than nurse-led care alone.12,26 In a Japanese stroke unit, embedding a full-time dentist to develop and lead an oral care system (including nurse training and standardized techniques) significantly lowered pneumonia incidence compared to periods without dentist involvement.14 These studies emphasize the importance of oral health care by dentists.

    The second reason was the lack of referral habits. Studies in Japan and the United States have highlighted the limited collaboration between hospitalists and dentists in routine clinical practice.27,28 Similarly, our study suggests that the lack of a habit of referring patients to dentists may be reflected in the low referral rates observed during aspiration pneumonia management. In contrast, a qualitative study from Germany indicated that while hospitalists did not perceive the need for collaboration with dentists, they considered such collaboration important.29 This suggests that the lack of collaboration between hospitalists and dentists may be driven by hospitalists’ attitudes. To address this challenge, implementing systematic referral prompts in electronic health records (EHR) may be beneficial. A study of oral health promotion in primary care found that automatically bundling dental referrals with related medical orders in EHR systems significantly improved referral consistency.30 One Japanese hospital’s program assigned a dentist to its acute stroke ward and defined procedures for nurses to request dental intervention promptly for any patient with compromised oral health.14 Additionally, borrowing core elements from antimicrobial stewardship programs, which are widely implemented to address antibiotic resistance, may be beneficial.31 These interventions reduce the referral barrier, potentially enhancing dental referral rates during aspiration pneumonia management.

    The third reason was the absence of dentists in hospitals. The proportion of dentists working in hospitals is as low as 0.4% and 3.0% in the United States and Japan, respectively.32,33 Furthermore, only about 28% of hospitals with dental departments have full-time dentists.16 This highlights the current shortage of hospital-based dentists capable of providing specialized oral health care. Teledentistry, a hub-and-spoke model in which intra-oral images and other clinical data are transmitted electronically from “spoke” wards to a central (hub) dentist, enables a university-hospital dentist to monitor and advise on the oral health of inpatients in surrounding hospitals even when no onsite dentist is available.34 Although there is limited direct evidence linking teledentistry to a reduction in the risk of aspiration pneumonia, it can enable hospitals to access dental care services remotely, potentially improving oral health management for at-risk patients.

    Background of Hospitalists Who Refer Patients to Dentists

    Several key characteristics were identified among hospitalists who referred patients to dentists. The first was the number of dentists in the hospital. Referral rates were higher in hospitals with dentists. Dentists have been reported to desire greater collaboration with their doctors.29 Our study found that hospitals with two or more dentists had higher referral rates than hospitals with no dentists or only one dentist. This suggests that hospitals with a larger dental workforce may be better equipped to provide a wider range of oral healthcare services, such as oral hygiene management and swallowing function support. The second factor was the presence of dental hygienists. Oral health care provided by dental hygienists has been shown to be effective in preventing aspiration pneumonia.35,36 Additionally, dental hygienists have been reported to play a key managerial role in medical-dental collaborations.37 The presence of dental hygienists may improve access to oral health care and facilitate collaboration between medical and dental professionals. The third factor was hospital type. Our findings indicate that hospitalists working in university hospitals had higher referral rates than those working in community-based hospitals. In Japan, university hospitals are legally required to offer 16 medical departments, including dentistry,38 whereas community-based hospitals have no such requirement. Consequently, university hospitals are guaranteed to have at least one dentist, which may have contributed to a more established referral habit. Additionally, university hospitals in Japan often treat patients with multiple underlying conditions, resulting in increased complexity and potentially higher referral rates. Moreover, because referrals to other specialties are more common for diseases outside one’s own expertise, this practice may have contributed to the higher referral rate observed in our study.39

    Actions Taken by Hospitalists Who Refer Patients for Dental Consultation

    A key finding of this study was the clear association between hospitalists’ attention to oral health in daily practice and the likelihood of referring patients to dentists. Hospitalists who routinely performed oral evaluations had higher referral rates than those who did not. Furthermore, among those who not only used the oral healthcare assessment tool but also confirmed whether patients had a primary care dentist, the referral rate exceeded 70%, suggesting that performing both actions as part of routine care further increased the likelihood of referral. Therefore, it is critical to conduct oral healthcare assessments. Paying attention to oral health can increase referral rates.

    The oral healthcare assessment tool is a screening method that evaluates oral health across eight categories and can be used for patients with dementia.40 Studies have shown that patients with Oral Health Assessment Tool (OHAT) scores of 3 or higher have significantly lower 60-day survival rates post-hospitalization compared to those with lower scores.41 Additionally, OHAT scores have been found to be significantly worse in patients with aspiration pneumonia compared to those with other types of community-acquired pneumonia.42 However, the utilization of OHAT remains low, with fewer than 5% of initial outpatient visits, including OHAT assessments.43 The findings of this study suggest that when hospitalists prioritize oral health using assessment tools and primary care dentists, they are more likely to refer patients to dentists. In Australian acute-care hospitals, national safety-and-quality guidance stipulates that an oral-health assessment, performed with a validated tool such as the OHAT, must be completed and documented within 24 hours of admission, alongside other vital signs.44 Consistent with earlier quality-improvement projects,30 embedding two mandatory EHR pop-up prompts at admission: (i) confirmation that the oral-health assessment is complete and (ii) documentation of whether the patient has a primary-care dentist, would ensure these checkpoints are actioned in real time. Designating these two assessments as mandatory Day-1 tasks within the admission order set is expected to expedite dental referrals and reduce the incidence of aspiration pneumonia.

    Limitations

    This study had some limitations. First, it focused on hospitalists in Japan, and the findings may not be generalizable to other countries with different healthcare systems and cultural backgrounds. Second, the survey targeted hospitalists primarily from general medicine departments, meaning that physicians from other medical specialties were excluded. This limitation should be considered when interpreting the results. Moreover, the questionnaire was designed by two of the authors and reviewed by the other authors. Because this is the original version, no external reference exists.

    Additionally, there is a possibility of selection bias, as the physicians who participated in this study may have had a greater interest in the research topic. Consequently, the perspectives of hospitalists with lower motivation to participate in such surveys may not have been adequately captured. Furthermore, because this study was based on a cross-sectional, self-report survey, it did not establish a causal relationship between actual clinical behavior and dental referrals. Nevertheless, there has never been a survey on medical-dental collaboration with a large number of respondents before, making this research an important baseline for future study.

    Future research should include a broader range of medical professionals, such as physicians from various specialties. Moreover, prospective studies are necessary to clarify the causal relationship between hospitalists’ actions and dental consultations.

    Conclusions

    This study investigated the status of collaboration between hospitalists and dentists in Japan regarding the management of aspiration pneumonia. The results revealed that organizational factors, such as the presence of a dental specialist, and individual factors, such as the degree to which an oral evaluation was performed, significantly affected the referral rates to dentists in aspiration pneumonia management. Attention to the oral environment during examinations may improve dental collaboration.

    Abbreviations

    JSHGM, Japanese Society of Hospital General Medicine; JPCA, Japanese Primary Care Association; OHAT, Oral Health Assessment Tool.

    Data Sharing Statement

    The datasets used and analyzed during the current study are available from the corresponding author upon reasonable request.

    Ethics Approval and Informed Consent

    The study was approved by the ethics committee of the Ashikaga Red Cross Hospital (No.2024-34) and was conducted in accordance with the principles of the Declaration of Helsinki. All participants reviewed the study document detailing data anonymization, voluntary participation, and the dissemination of research results prior to participation. Only participants who provided informed consent (opt-in) were included in the study. Additionally, the participants could withdraw from the study at any time.

    Author Contributions

    All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising, or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.

    Funding

    This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.

    Disclosure

    The authors report no conflicts of interest in this work.

    References

    1. United Nations. 2019. World population prospects. Available from: https://population.un.org/wpp/. Accessed February 21, 2025.

    2. Arai H, Ouchi Y, Toba K, et al. Japan as the front-runner of super-aged societies: perspectives from medicine and medical care in Japan. Geriatr Gerontol Int. 2015;15(6):673–687. doi:10.1111/ggi.12450

    3. Tomonaga I, Koseki H, Imai C, et al. Incidence and characteristics of aspiration pneumonia in the Nagasaki Prefecture from 2005 to 2019. BMC Pulm Med. 2024;24(1):191. doi:10.1186/s12890-024-03015-8

    4. Gupte T, Knack A, Cramer JD. Mortality from aspiration pneumonia: incidence, trends, and risk factors. Dysphagia. 2022;37(6):14931500. doi:10.1007/s00455-022-10412-w

    5. Wu CP, Chen YW, Wang MJ, Pinelis E. National trends in admission for aspiration pneumonia in the United States, 2002-2012. Ann Am Thorac Soc. 2017;14(6):874–879. doi:10.1513/AnnalsATS.201611-867OC

    6. Ocrospoma S, Restrepo MI. Severe aspiration pneumonia in the elderly. J Intensive Med. 2024;4(3):307–317. doi:10.1016/j.jointm.2023.12.009

    7. Uno I, Kubo T. Risk factors for aspiration pneumonia among elderly patients in a community-based integrated care unit: a retrospective cohort study. Geriatrics. 2021;6(4):113. doi:10.3390/geriatrics6040113

    8. Abdelaziz AA, Doghish AS, Salah AN, et al. (When oral health affects overall health: biofilms, dental infections, and emerging antimicrobial strategies. Infection. 2025. doi:10.1007/s15010-025-02533-9

    9. Khadka S, Khan S, King A, Goldberg LR, Crocombe L, Bettiol S. Poor oral hygiene, oral microorganisms and aspiration pneumonia risk in older people in residential aged care: a systematic review. Age Ageing. 2021;50(1):81–87. doi:10.1093/ageing/afaa102

    10. Son M, Jo S, Lee JS, Lee DH. Association between oral health and incidence of pneumonia: a population-based cohort study from Korea. Sci Rep. 2020;10(1):9576. doi:10.1038/s41598-020-66312-2

    11. Soutome S, Yanamoto S, Funahara M, et al. Preventive effect on post-operative pneumonia of oral health care among patients who undergo esophageal resection: a multi-center retrospective study. Surg Infect. 2016;17(4):479–484. doi:10.1089/sur.2015.158

    12. Miyagami T, Nishizaki Y, Imada R, et al. Dental care to reduce aspiration pneumonia recurrence: a prospective cohort study. Int Dent J. 2024;74(4):816–822. doi:10.1016/j.identj.2023.11.010

    13. van der Maarel-Wierink CD, Vanobbergen JN, Bronkhorst EM, Schols JM, de Baat C. Oral Health care and aspiration pneumonia in frail older people: a systematic literature review. Gerodontology. 2013;30(1):3–9. doi:10.1111/j.1741-2358.2012.00637.x

    14. Ozaki K, Teranaka S, Tohara H, Minakuchi S, Komatsumoto S. Oral management by a full-time resident dentist in the hospital ward reduces the incidence of pneumonia in patients with acute stroke. Int J Dent. 2022;2022:6193818. doi:10.1155/2022/6193818

    15. Chan AKY, Tsang YC, Jiang CM, Leung KCM, Ecm L, Chu CH. Integration of oral health into general health services for older adults. Geriatrics. 2023;8(1):20. doi:10.3390/geriatrics8010020

    16. Medical Division. HIB, Ministry of Health, Labour and Welfare. Summary of Revision of Medical Service Fee in Reiwa 4 years (Dentistry). Available from: https://www.mhlw.go.jp/content/12400000/000922373.pdf. Accessed January 31, 2025.

    17. Miyagami T, Teranaka S, Mine Y, et al. Lack of physician-dentist collaboration in aspiration pneumonia prevention. Int J Gen Med. 2024;17:1293–1295. doi:10.2147/IJGM.S405712

    18. Hamada O, Tsutsumi T, Tsunemitsu A, Fukui T, Shimokawa T, Imanaka Y. Impact of the hospitalist system in Japan on the quality of care and healthcare economics. Intern Med. 2019;58(23):3385–3391. doi:10.2169/internalmedicine.2872-19

    19. Peterson MC. A systematic review of outcomes and quality measures in adult patients cared for by hospitalists vs nonhospitalists. Mayo Clin Proc. 2009;84(3):248–254. doi:10.4065/84.3.248

    20. Rifkin WD, Conner D, Silver A, Eichorn A. Comparison of processes and outcomes of pneumonia care between hospitalists and community-based primary care physicians. Mayo Clin Proc. 2002;77(10):1053–1058. doi:10.4065/77.10.1053

    21. Miyagami T, Shimizu T, Kosugi S, et al. Roles considered important for hospitalist and non-hospitalist generalist practice in Japan: a survey study. BMC Prim Care. 2023;24(1):139. doi:10.1186/s12875-023-02090-w

    22. Ministry of Health Labour and Welfare. Summary of the 2023 medical facilities (Movement) survey and hospital report. Available from: https://www.mhlw.go.jp/toukei/saikin/hw/iryosd/23/. Accessed January 31, 2025.

    23. Ishikawa A, Yoneyama T, Hirota K, Miyake Y, Miyatake K. Professional oral health care reduces the number of oropharyngeal bacteria. J Dent Res. 2008;87(6):594–598. doi:10.1177/154405910808700602

    24. Terpenning M. Geriatric oral health and pneumonia risk. Clin Infect Dis. 2005;40(12):1807–1810. doi:10.1086/430603

    25. Nakamura T, Kurosaki S. Effects of early dysphagia rehabilitation by speech-language-hearing therapists on patients with severe aspiration pneumonia. Prog Rehabil Med. 2020;5:20200020. doi:10.2490/prm.20200020

    26. Haresaku S, Aoki H, Kubota K, et al. Nurses’ perceptions of Oral Health care provision after the COVID-19 lockdown. Int Dent J. 2022;72(2):242–248. doi:10.1016/j.identj.2021.06.004

    27. Wakabayashi H. Medical-dental collaboration in general and family medicine. J Gen Fam Med. 2019;20(2):47. doi:10.1002/jgf2.237

    28. Shimpi N, Schroeder D, Kilsdonk J, et al. Medical providers’ Oral Health knowledgeability, attitudes, and practice behaviors: an opportunity for interprofessional collaboration. J Evid Based Dent Pract. 2016;16(1):19–29. doi:10.1016/j.jebdp.2016.01.002

    29. Sippli K, Rieger MA, Huettig F. GPs’ and dentists’ experiences and expectations of interprofessional collaboration: findings from a qualitative study in Germany. BMC Health Serv Res. 2017;17(1):179. doi:10.1186/s12913-017-2116-4

    30. Mitchell-Royston L, Nowak A, Silverman J. Interprofessional Study of Oral Health in Primary Care: Final Report. Chicago (IL): American Academy of Pediatric Dentistry, Pediatric Oral Health and Policy Research Center; 2014.

    31. Salah A, El-Housseiny G, Elleboudy N, Yassien M. Antimicrobial stewardship programs: a review. Arch Pharmaceut Sci Ain Shams Univ. 2021;5(1):143–157. doi:10.21608/aps.2021.76105.1059

    32. Institute of Medicine. Advancing Oral Health in America. Washington, DC: National Academies Press; 2011.

    33. Ministry of Health. LaW: statistics of physicians, dentists and pharmacists. Available from: https://www.mhlw.go.jp/toukei/list/33-20.html. Accessed January 22, 2025.

    34. Langelier M, Rodat C, Moore J. Case Studies of 6 Teledentistry Programs: Strategies to Increase Access to General and Specialty Dental Services. Rensselaer, NY: Oral Health Workforce Research Center, Center for Health Workforce Studies, School of Public Health, SUNY Albany; 2016.

    35. Adachi M, Ishihara K, Abe S, Okuda K. Professional oral health care by dental hygienists reduced respiratory infections in elderly persons requiring nursing care. Int J Dent Hyg. 2007;5(2):69–74. doi:10.1111/j.1601-5037.2007.00233.x

    36. Hama K, Iwasa Y, Ohara Y, et al. Pneumonia incidence and oral health management by dental hygienists in long-term care facilities: a 1-year prospective multicentre cohort study. Gerodontology. 2022;39(4):374–383. doi:10.1111/ger.12604

    37. Nakayama R, Soga Y, Fujii S, et al. Expanding on the professional role of dental hygienists as key managers of medical-dental and hospital-dental clinic collaboration in a local Japanese hospital without a dentistry department: from a questionnaire survey after a web seminar. Int J Dent Hyg. 2023;21(3):533–540. doi:10.1111/idh.12684

    38. Ministry of Health. LaW: outline of advanced treatment hospital. Available from: https://www.mhlw.go.jp/file/05-Shingikai-10801000-Iseikyoku-Soumuka/0000087302.pdf. Accessed January 31, 2025.

    39. Nishizaki Y, Nozawa K, Shinozaki T, et al. Difference in the general medicine in-training examination score between community-based hospitals and university hospitals: a cross-sectional study based on 15,188 Japanese resident physicians. BMC Med Educ. 2021;21(1):214. doi:10.1186/s12909-021-02649-0

    40. Chalmers JM, King PL, Spencer AJ, Wright FA, Carter KD. The oral health assessment tool—validity and reliability. Aust Dent J. 2005;50(3):191–199. doi:10.1111/j.1834-7819.2005.tb00360.x

    41. Maeda K, Mori N. Poor oral health and mortality in geriatric patients admitted to an acute hospital: an observational study. BMC Geriatr. 2020;20(1):26. doi:10.1186/s12877-020-1429-z

    42. Nishizawa T, Niikura Y, Akasaka K, et al. Pilot study for risk assessment of aspiration pneumonia based on oral bacteria levels and serum biomarkers. BMC Infect Dis. 2019;19(1):761. doi:10.1186/s12879-019-4327-2

    43. Scherr S, Idzik S, Williams D. Integrating oral health screening into primary care to promote dental referrals in Maryland. J Dr Nurs Pract. 2020;13(1):42–63. doi:10.1891/2380-9418.JDNP-D-19-00041

    44. Australian Commission on Safety and Quality in Health Care. Oral Health Care for Adult Inpatients: recommendations. Sydney: ACSQHC; 2023. Available from: https://www.safetyandquality.gov.au/publications-and-resources/resource-library/oral-health-care-adult-inpatients-recommendations. Accessed June 24, 2025.

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  • The global burden of motor neuron disease: a systematic and additional analysis of global burden disease study 2021 | Orphanet Journal of Rare Diseases

    The global burden of motor neuron disease: a systematic and additional analysis of global burden disease study 2021 | Orphanet Journal of Rare Diseases

    Data source

    The GBD 2021 employed the most up-to-date epidemiological data, complemented by refined and standardized methodologies, to systematically and comprehensively quantify health losses across 369 diseases and injuries, as well as 87 risk factors, stratified by age, sex, and geographical location, encompassing 204 countries and territories. The GBD team is committed to annual updates to ensure the accuracy and relevance of their estimates [12]. The intricacies of the methodologies applied within GBD 2021 have been thoroughly documented in prior publications [13].

    To address data gaps and ensure smoothness across age, time, and location, the collected data underwent modeling via spatiotemporal Gaussian process regression. This approach facilitated interpolation in regions with incomplete datasets. Furthermore, to correct for biases stemming from diverse case definitions and study methodologies across regions, a meta-regression framework incorporating Bayesian priors, regularization, and trimming techniques was employed.

    From GBD 2021, we extracted estimates and their corresponding 95% uncertainty intervals (UIs) for incidence, deaths, prevalence, and DALYs attributed to MND. All rates reported herein are standardized to per 10,000 population. Additionally, the sociodemographic index (SDI), a composite indicator reflecting income, education, and fertility levels, serving as a proxy for sociodemographic development, was utilized to categorize the 204 countries and territories into five distinct groups: high, high-middle, middle, low-middle, and low, as defined by their SDI values [14].

    Descriptive analysis

    To gain a holistic understanding of the burden of MND, we conducted descriptive analyses at the global, regional, and national levels. Specifically, we visually presented the global trends in the number of cases, crude rate, and age-standardized rate (ASR) for incidence, deaths, prevalence, and DALYs related to MND, disaggregated by sex (both sexes, males, and females) and spanning the period from 1990 to 2021. Furthermore, we compared the number of cases and ASR for the aforementioned indicators across global, regional (comprising 54 GBD geographic regions), and national (encompassing 204 countries and territories) levels, as well as within the five SDI groups.

    Trend analysis

    In the Trend Analysis section, we initially employed the Estimated Annual Percentage Change (EAPC) to quantify the overarching trend in the burden of MND. Given the importance of standardization when comparing diverse groups with varying age structures or a single group experiencing temporal changes in its age profile, the EAPC-measured trend of the ASR emerges as a more robust metric for monitoring shifts in disease patterns [15]. To derive this metric, we constructed a linear regression model where the natural logarithm of the ASR (ln(ASR)) served as the dependent variable (y), and the calendar year acted as the independent variable (x). Subsequently, the EAPC was calculated using the formula (exp(β)-1) * 100%, with its 95% confidence interval (CI) also being extracted from the model [16]. In interpreting the EAPC estimates, if both the EAPC value and the lower bound of its 95% CI are greater than 0, the ASR is deemed to be in an increasing trend. Conversely, if both the EAPC value and the upper bound of its 95% CI are less than 0, the ASR is considered to be in a decreasing trend. In all other cases, the ASR is classified as stable. This approach ensures a rigorous and standardized methodology for assessing temporal trends in the ASR of MND.

    Furthermore, we used age-period-cohort (APC) model to explore the underlying trends in DALYs stratified by age, period, and birth cohort. Typically, the APC model fits a log-linear Poisson model on the Lexis diagram of observed rates and quantifies the additional effects of age, period, and birth cohort. The methodological details of APC model are described in previous literature [17]. The multicollinearity between age, period, and birth cohort inevitably leads to identification issues, making it difficult to estimate the unique effects of each age, period, and birth cohort. To address this issue, the intrinsic estimator (IE) algorithm was used to estimate the coefficients of the APC model. This study employed the IE to solve the APC model, where coefficients greater than 0 indicate increased risk, and those less than 0 indicate decreased risk. The effect coefficients were transformed into natural logarithms to calculate the relative risk (RR), enabling the observation of the effects of age, period, and cohort on MND DALYs trends. The DALYs for MND and population data of each country or region were served as data input for APC model. The data was re-coded into consecutive six 5-year periods (1990–1994, 1995–1999, … , 2015–2019), consecutive 5-year age groups (0–4, 5–9, … , 90–94, 95 plus), consecutive 5-year birth cohorts (1895–1899, 1900–1904, … , 2015–2019) to estimate the overall temporal trend in incidence, prevalence, deaths, and DALYs.

    Cross-country inequality analysis

    To ensure evidence-based health planning, we conducted a comprehensive cross-country inequality analysis aimed at monitoring health disparities. Specifically, we employed the Slope Index of Inequality (SII) as a key metric, which was derived from regressing the country-level prevalence of the disease across all age groups against a relative position scale tied to sociodemographic development. To account for heteroscedasticity, a robust linear regression model was applied. This method utilizes iteratively reweighted least squares, giving smaller weights to observations with larger residuals, thus minimizing the influence of outliers and ensuring more stable and reliable trend estimates [18]. This approach allowed us to quantify inequalities in MND at global level and across 21 GBD regions.

    Furthermore, we calculated the Health Inequality Concentration Index by numerically integrating the area beneath the Lorenz Concentration Curve. This curve was meticulously fitted using the cumulative relative distribution of populations, ordered by their SDI, and the corresponding incidence, prevalence, deaths, and DALYs attributable to the disease [19, 20]. This methodology provided a robust assessment of the concentration of health burden across nations, enabling us to identify disparities and inform targeted interventions. A negative SII/concentration index indicates that as SDI increases, ASDR decreases, and vice versa. The greater the absolute value of the SII/concentration index, the greater the degree of inequality. Their inequality value and implications are presented in Table 1.

    Table 1 The changing pattern of inequalities from 1990 to 2021 and their implications [21]

    Decomposition analysis

    To gain a profound understanding of the explanatory factors underpinning the variations in MND incidence, prevalence, deaths, and DALYs from 1990 to 2021, we performed a comprehensive decomposition analysis. This analysis dissected the contributions of population size, age structure, and epidemiological changes to the observed trends [22, 23]. By disentangling these components, we aimed to quantify the specific impact of each factor on the evolution of MND burden over time.

    The decomposition methodology enabled us to estimate the number of incidence cases, prevalent cases, deaths, and DALYs attributable to each factor at every location under consideration. The calculation of these metrics for each component was carried out as follows:

    ({X_{ay,py,ey}} = sumnolimits_{i = 1}^{20} {left( {{a_{i,y}} * {p_y} * {e_{i,y}}} right)} )(X = incidence, prevalence, deaths and DALYs)

    Where the ({X_{ay,py,ey}}) represented X based on the factors of age structure, population, and specific year (y); ({a_{i,y}}) represented the proportion of population for the age category (i) of the 20 age categories in year (y); ({p_{y}}) represented the total population in year (y) and ({e_{i,y}}) represented X rate for the age category (i) in year (y).

    The contribution of each factor to the change in incidence, prevalence, deaths and DALYs from 1990 to 2021 was defined by the effect of one factor changing while the other factors were held constant.

    Predictive analysis

    To inform the formulation of effective public health policies and the optimal allocation of healthcare resources, we conducted a predictive analysis of the MND burden in the coming decades. For this purpose, we employed the Bayesian age-period-cohort (BAPC) model, augmented with the integrated nested Laplace approximation (INLA) technique. This advanced approach, which has been shown to outperform the conventional annual percentage change model in terms of coverage and precision, was utilized to forecast the global MND burden until 2046.

    The adoption of INLA within the BAPC framework offers several advantages. By approximating marginal posterior distributions, it mitigates the mixing and convergence issues that are often encountered with the Markov Chain Monte Carlo sampling techniques traditionally applied in Bayesian methods [24]. This enhancement ensures more reliable and accurate predictions of the future MND burden, thereby supporting evidence-based decision-making in public health planning.

    Frontier analysis

    To assess the interplay between the burden of MND and sociodemographic development, we employed a frontier analysis approach. This methodology aimed to delineate the lowest potentially attainable ASR of incidence, prevalence, deaths, and DALYs for each country or territory, contingent upon its SDI. The frontier serves as a benchmark, indicating the minimal achievable level given a country’s or territory’s development status. The deviation from this frontier, termed the effective difference, highlights potential untapped opportunities for improvement or gains, commensurate with the country’s or territory’s position on the development spectrum.

    To accommodate non-linear relationships, we conducted a data envelope analysis utilizing the free disposal hull method. This analysis generated an age-adjusted frontier by SDI, utilizing data spanning from 1990 to 2021 [25]. To account for uncertainty, we implemented a bootstrapping procedure, drawing 1,000 samples with replacement from the entire dataset encompassing all countries and territories across all years. From these bootstrapped samples, we computed the mean incidence, prevalence, deaths, and DALYs at each SDI value.

    Subsequently, we employed LOESS (Locally Estimated Scatterplot Smoothing) regression with a local polynomial degree of 1 and a span of 0.2 to produce a smooth frontier line [25]. This approach ensured a robust and visually interpretable representation of the frontier, while mitigating the influence of outliers. To further refine the analysis, super-efficient countries, which may distort the frontier due to exceptional performance, were excluded from the frontier generation process.

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  • Evaluation of intraductal carcinoma and invasive cribriform carcinoma as predictors of genetic mutations in systemic treatment-naïve prostate cancer patients | BMC Cancer

    Evaluation of intraductal carcinoma and invasive cribriform carcinoma as predictors of genetic mutations in systemic treatment-naïve prostate cancer patients | BMC Cancer

    The primary aim of this study was to investigate the relationship between IDC-P/ICC and HRR mutations in a real-world cohort of 347 patients with prostate cancer. We further assessed the association between IDC-P/ICC and other molecular alterations, including MMR status, MSI, and TMB. Our study revealed that the presence of IDC-P/ICC was not associated with HRR mutations or MMR status. Notably, mutations in HRR were linked to a younger age and a higher Grade Group, yet they showed no correlation with IDC-P/ICC status or metastatic stage.

    IDC-P and ICC are histologically distinct entities but exhibit similarly aggressive clinical courses, both being associated with advanced disease stages, recurrence, and poor outcomes [24, 25]. Although the 2019 ISUP consensus recommends that IDC-P and ICC be reported separately due to their distinct diagnostic and prognostic implications, in real-world practice, this distinction can be challenging because of overlapping morphologic features [21, 26]. Accordingly, the NCCN guidelines recommend germline testing for patients with either IDC-P or cribriform morphology, reflecting their shared clinical significance and association with genetic mutations [11]. At our institution, consistent with contemporaneous diagnostic practice during the study period, IDC-P and ICC were evaluated morphologically without the use of routine basal cell immunohistochemistry and were reported together rather than separately. While this approach reflects actual clinical practice, it differs from current consensus recommendations and may have introduced variability in histologic classification, which should be considered a limitation of our study.

    Additionally, our data revealed discrepancies between biopsy and prostatectomy findings. While most IDC-P/ICC cases were detected on biopsy, approximately 13% were identified only on prostatectomy specimens, and one case was positive on biopsy but negative on prostatectomy, suggesting sampling variation or intratumoral heterogeneity. These findings align with prior studies showing the limited sensitivity of biopsy. Ericson et al. [27] reported a sensitivity of 56.5% with no added benefit from MRI fusion, and Masoomian et al. [28] found that biopsy detected IDC-P/ICC in only 26.9% of cases compared with 51.8% at prostatectomy, corresponding to a sensitivity of 47.2%. More recently, Bernardino et al. demonstrated that false-negative biopsies were associated with higher pathological stage and increased risk of biochemical recurrence [29]. Taken together, these results highlight that prostate biopsy has limited sensitivity for IDC-P/ICC detection, as reflected by the 13% underdetection rate in our cohort. This reinforces that biopsy IDC-P/ICC status alone is insufficient to guide genetic testing and underscores the need to integrate additional clinical and pathological factors.

    Contrary to previous studies suggesting a link between IDC-P/ICC and HRR mutations [7,8,9], our multivariable analysis revealed that IDC-P/ICC is not a significant predictor of HRR mutation status. These findings are in line with those of recent studies [30, 31]. Mahlow et al. [31] concluded that pathologic patterns alone are insufficient to predict HRR mutations in advanced prostate cancer. While their study included only six IDC-P cases, our larger cohort of 254 patients with IDC-P/ICC provides robust support for their conclusions. Lozano et al. [30] found no significant differences in IDC-P/ICC between germline BRCA2 (gBRCA2) carriers and non-carriers. However, they discovered that bi-allelic BRCA2 loss in primary prostate tumors was independently associated with both IDC-P and cribriform morphologies. They proposed that tumors with a gBRCA2 mutation and intact second allele may preserve some BRCA2 function, potentially preventing these histologies. Our study extends these findings by examining a broader spectrum of HRR genes, showing that somatic HRR mutation status is not indicated by the presence of IDC-P/ICC. Since our cohort primarily consisted of metastatic cases, it is important to note that while some non-metastatic patients were included, further studies are required to fully understand the role of IDC-P/ICC in localized prostate cancer. With HRR gene mutation prevalence around 25% in metastatic disease [32] but less than 10% in localized disease [33], the consistent lack of association between IDC-P/ICC and HRR mutations across studies raises questions about the appropriateness of relying on cribriform pattern status as a trigger for genetic testing, particularly as outlined in current NCCN guidelines [11].

    Another potential explanation for the discrepancy between IDC-P/ICC and HRR mutations lies in our limited understanding of IDC-P pathogenesis and its underlying molecular events. The pathogenesis of IDC-P can vary, with most cases resulting from retrograde spreading of adjacent aggressive prostate cancer into ducts, but de novo IDC-P can also occur [2]. These different origins might contribute to the observed variability in HRR gene alterations in IDC-P/ICC. The heterogeneity in IDC-P origins could explain why our study and others have found inconsistent associations between IDC-P/ICC and not only HRR but also other mutations. Contrary to previous studies’ results [9, 10], our data do not show a statistical difference in MMR mutations between IDC-P/ICC positive and negative groups (2.8% vs. 1.1%, P = 0.687). Additionally, another potential poor prognostic marker, MSI-high status, was also not significantly different between the two groups (1.1% vs. 1.2%, P > 0.999). These findings prompt us to reconsider the use of IDC-P/ICC as a sole indicator for genetic testing and suggest that a combination of clinical and pathological factors may provide better stratification for identifying patients who would benefit from comprehensive genomic profiling. Given the relatively low prevalence of IDC-P/ICC, future research would benefit from multicenter collaborations with centralized pathologic review of specimens for genetic testing. Such large-scale efforts could provide more definitive insights into the relationship between IDC-P/ICC and molecular alterations.

    HRR mutations occur not only in metastatic disease but also in locally advanced or regional diseases. Our study found that HRR gene mutations are associated with high Grade Groups and younger age at diagnosis, regardless of metastatic status. While current guidelines focus on metastatic disease for recommending genetic testing, emerging evidence suggests HRR status may provide valuable prognostic information even in localized disease [22]. With PARP inhibitors now showing efficacy in metastatic prostate cancer [13,14,15,16], there is potential for their application in localized disease as well [34]. This evolving landscape underscores the need for a more stratified approach to genetic testing in prostate cancer. Based on our study, a combination of younger age and higher Grade Group may be good indicators for genetic testing, rather than relying solely on a single morphological factor.

    The major limitations of this study are its retrospective, single-center design, which may limit the generalizability of our findings to broader populations. Additionally, the lack of a standardized NGS testing protocol introduces potential selection bias, as genomic testing was performed at the discretion of the treating physician and was most often applied to patients with aggressive clinicopathological features, rather than uniformly across all prostate cancer cases. Furthermore, as we collected somatic NGS test data, the results cannot be directly applied to germline findings. Another limitation is that IDC-P and ICC were assessed morphologically without routine basal cell immunohistochemistry and were not reported separately, which may have introduced variability in classification. Despite these limitations, our study provides real-world evidence of the correlation of IDC-P/ICC with HRR, MMR, MSI, and TMB, which are important predictors in the era of molecularly targeted systemic treatments.

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  • Development and characterization of a chitosan-stabilized pickering emulsion incorporating thyme essential oil to extend the shelf life of strawberries during cold storage | BMC Plant Biology

    Development and characterization of a chitosan-stabilized pickering emulsion incorporating thyme essential oil to extend the shelf life of strawberries during cold storage | BMC Plant Biology

    Nanogel chitosan characterization

    Fourier-transform infrared spectroscopy (FTIR)

    In this study, the nanogel was initially synthesized using modified chitosan. To achieve this modification, a portion of the free amine groups in chitosan react with stearate esters to form amide bonds, which can enhance the properties of chitosan, such as its solubility and bioactivity via the intermediate coupling agent 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC). The conjugation was performed at different molar ratios to assess the reaction efficiency. To confirm the formation of a chemical bond between the amine groups of chitosan and the carboxyl groups of stearate ester, Fourier Transform Infrared Spectroscopy (FT-IR) was employed. Figure 1 presents the FT-IR spectra of pure chitosan, stearate ester, and the resulting nanogels synthesized through the chitosan-stearate ester linkage.

    Figure 1a presents the FT-IR spectrum of chitosan. A broad absorption band is observed in the range of 3400–3500 cm⁻¹, which is attributed to the hydroxyl (–OH) groups present in the chitosan structure. At around 1580 cm⁻¹, a peak appears that corresponds to bending vibrations of amide and amine bonds. The peak observed at 2924 cm⁻¹ is related to C–H stretching vibrations, indicating the presence of methyl (–CH₃) and methylene (–CH₂) groups in the chitosan backbone. A distinct peak at 1634 cm⁻¹ is assigned to the stretching vibrations of the amide carbonyl group (C = O), which is associated with the non-deacetylated amine groups. In the region around 1157 cm⁻¹, an asymmetric stretching band related to the ether group (C–O–C) is detected, which is a characteristic feature of chitosan’s molecular structure. Finally, a band at 1064 cm⁻¹ is observed, representing the C–O stretching vibrations, which are also commonly found in the chitosan structure. These findings are consistent with previous reports by Chiono et al. and Peng et al. [24, 25].

    Figure 1b displays the FT-IR spectrum of stearate ester. In this spectrum, distinct peaks appear at 2917 cm⁻¹ and 2849 cm⁻¹, which correspond to the stretching vibrations of C–H bonds. The peak observed at 1701 cm⁻¹ is attributed to the stretching vibration of the carbonyl group (C = O) present in the stearate ester structure. Additionally, a peak at 1470 cm⁻¹ is associated with the bending vibration of C–H bonds. In the region of 1261 cm⁻¹, a peak corresponding to the stretching vibrations of the C–O group is observed. The peak at 940 cm⁻¹ indicates the bending vibrations of hydroxyl groups (O–H). Finally, two additional peaks at 721 cm⁻¹ and 686 cm⁻¹ are related to the bending vibrations of C–H bonds within the long hydrocarbon chain of stearate ester.

    Figure 1c presents the FT-IR spectrum of the chitosan–stearate ester nanogel. When compared to the spectrum of pure chitosan, notable changes in the intensity of several peaks can be observed. Specifically, the peak intensity around 1630 cm⁻¹, which corresponds to the carbonyl (C = O) group, is markedly increased in the chitosan–stearate ester nanogel. In contrast, the peak at 1525 cm⁻¹, attributed to the free amine (–NH₂) groups, also exhibits changes in intensity. The increased ratio of the C = O peak intensity to that of NH₂ suggests the formation of a higher number of amide bonds in the final nanogel structure. These alterations in peak intensities provide strong evidence for a successful reaction between the amine groups of chitosan and the carboxyl groups of stearate ester. Similar findings have been reported in previous studies, including those conducted by Guo et al., Hadian et al., Waldron et al., and Wang et al. [26,27,28,29].

    Fig. 1

    FTIR spectra of chitosan a stearate ester b and nongel-chitosan/stearate ester c

    SEM analysis of chitosan–stearate ester nanogel

    Figure 2 displays the scanning electron microscopy (SEM) image of the nanogels synthesized from chitosan and stearate ester. According to this image, the nanogels exhibit a predominantly spherical morphology, with particle sizes estimated to be less than 100 nanometers. This spherical structure forms when the hydrophobic fatty acid stearate ester is incorporated into the chitosan polymer. In a polar medium, this combination tends to self-assemble into micelle-like spherical structures, where the hydrophobic tails orient inward while the hydrophilic heads face outward.

    Fig. 2
    figure 2

    The scanning electron microscopy (SEM) image of the chitosan– stearate ester nanogel containing thyme essential oil

    As a biocompatible polymer, chitosan does not trigger allergic responses, and its metabolic byproducts, including amino sugars, are non-toxic and fully absorbed by the body. Numerous studies have explored chitosan-based nanogels formulated with various cross-linking agents. For instance, Nasti et al. developed chitosan nanoparticles with sizes ranging from 160 to 260 nm using tripolyphosphate as a cross-linker [30]. In another study, Lee et al. prepared nanogels of approximately 200 nm in size from a combination of chitosan and glycol [31]. They emphasized that achieving a balance between hydrophobic and hydrophilic interactions is critical for micelle formation, which serves as the primary mechanism behind the nanostructure’s self-assembly. Previous findings have demonstrated that reducing particle size enhances the efficiency of coating materials. Moreover, the use of smaller particles in fruit coatings leads to a reduction in the diameter of surface stomatal pores. This, in turn, decreases the permeability to oxygen and water vapor, effectively limiting enzymatic activity and delaying the onset of fruit spoilage [32].

    TEO release from chitosan-based nanogel encapsulation

    Evaluating the release profile of encapsulated essential oil is critical to determine whether the chitosan nanogel delivers thymol essential oil in a rapid (burst release) or gradual (sustained release) manner to the fruit matrix. To this end, a spectrophotometric method was employed to monitor the release of thymol essential oil from the chitosan capsules. The release behavior of thymol essential oil from chitosan–stearate ester nanogels under ambient temperature conditions (as shown in Fig. 3) exhibited a biphasic pattern. An initial burst release phase was observed up to day 2, followed by a sustained release phase lasting until day 10. Subsequently, a second burst release phase occurred until day 16, after which a second sustained release phase continued until the end of day 28.

    Fig. 3
    figure 3

    Kinetics of thyme essential oil (TEO) release from chitosan– stearate ester nanogels

    The nanogel developed in this study, due to its amphiphilic structure comprising polar (chitosan region) and nonpolar (stearate ester chains) domains, provided an appropriate matrix for the gradual release of the hydrophobic constituents of thyme essential oil into the surrounding environment. This structural characteristic facilitated the slow and controlled release of the active compounds. These findings align with Mohammadi et al. [33], who demonstrated the controlled release of Shirazi thyme essential oil from encapsulated delivery systems. Similarly, Abdalla et al. [34] observed a biphasic release pattern for lemon essential oil encapsulated in pectin–chitosan matrices, consisting of an initial burst followed by sustained release.

    Weight loss

    Statistical analysis revealed significant differences (p < 0.05) in fruit weight loss during the storage period as influenced by the applied treatments. Although weight loss was observed across all treatments and the control over time, the control group exhibited a markedly higher reduction from day 8 onward compared to the treated samples. All treatment groups effectively delayed weight loss throughout the storage period (Fig. 4).

    Fig. 4
    figure 4

    Effects of encapsulated thyme essential oil on the weight loss of strawberry fruits during 16 days of storage at 4 °C. Data are presented as means ± standard deviation (n = 3). Treatments: C (control), E1 (thyme essential oil coating, 600 µL/L), E2 (thyme essential oil coating, 1200 µL/L), NE1 (encapsulated thyme essential oil coating, 600 µL/L), and NE2 (encapsulated thyme essential oil coating, 1200 µL/L)

    Interestingly, the application of the essential oil coating at lower concentrations resulted in a more favorable reduction in weight loss. In contrast, higher concentrations of the free essential oil led to increased weight loss. However, encapsulation of the essential oil using chitosan altered this trend; the highest concentration of encapsulated essential oil resulted in the lowest weight loss during storage. By the end of the storage period (day 16), the control fruits had lost 19.66% of their initial weight, while the corresponding weight loss for the E1, E2, NE1, and NE2 treatments was 10.03%, 11.83%, 9.06%, and 7.8%, respectively.

    The primary causes of fruit weight loss appear to be moisture transfer from the fruit tissue to the surrounding environment [35] and increased respiration intensity [36]. Additionally, the reduced weight loss observed in fruits coated with thyme essential oil may be attributed to stomatal blockage, which ultimately slows down active metabolic processes and respiration rates. The decrease in weight loss in thyme oil-treated fruits may also result from the semi-permeable nature of the thyme oil coating, which affects moisture loss, respiration, and solute transport across membranes [37]. From day 8 onward, treatments containing higher concentrations of free essential oil exhibited increased weight loss, possibly due to greater evaporation of the essential oil over prolonged storage. The volatile nature of the essential oil may alter the fruit’s tissue structure and molecular integrity, a phenomenon previously reported during investigations of different concentrations of thyme essential oil on strawberry shelf life [37]. Encapsulation of thyme essential oil within chitosan significantly reduced its evaporation rate and prolonged its effectiveness in preserving tissue moisture content by limiting gas exchange between the atmosphere and the fruit tissue. Consequently, unlike the free essential oil treatment at higher concentration (E2), the most pronounced effect was observed in the corresponding encapsulated treatment (NE2). Reduced water loss following the application of plant essential oils and nano-encapsulated formulations has also been reported in sweet marjoram [38], as well as in other horticultural crops such as strawberries [39], wine grapes [40], and bell peppers [37, 41]. One of the advantages of using nanoparticles lies in their high surface-area-to-volume ratio, which enhances reactivity and cellular penetration [42]. Therefore, nanoencapsulation of thyme essential oil using chitosan in the present study effectively contributed to preserving the fresh weight of strawberries over the storage period.

    Firmness

    The effect of thyme essential oil (E1 and E2) and nano-encapsulated thyme essential oil (NE1 and NE2) in chitosan-based nanogels on the firmness of strawberry fruit during storage is presented in Fig. 5. Regardless of the treatment type, fruit firmness declined over the storage period, decreasing from an initial value of 2.66 N at the beginning of the experiment. Over the 16-day storage period, the reduction in firmness for the coating treatments E1, E2, NE1, and NE2 was 58.47%, 37%, 23%, and 9.06%, respectively. In contrast, the control group exhibited a significantly greater firmness loss of 66%, particularly when compared to the NE2 treatments. The highest firmness retention was observed in the NE2 treatment (2.03 N), followed by NE1 (1.66 N).

    Fig. 5
    figure 5

    Effects of encapsulated thyme essential oil on the firmness of strawberry fruits during 16 days of storage at 4 °C. Data were represented as means ± standard deviation (n = 3). Treatments: C (control), E1 (thyme essential oil coating 600 µL/L), E2 (thyme essential oil coating 1200 µL/L), NE1 (encapsulated thyme essential oil coating 600 µL/L) and NE2 (encapsulated thyme essential oil coating 1200 µL/L)

    The notable reduction in firmness is primarily attributed to tissue water loss and the activation of cell wall–degrading enzymes such as pectin methyl esterase and polygalacturonase [43]. The reduced weight loss observed in the TEO and NE-TEO treatments highlights the efficacy of these coatings in limiting moisture loss and consequently slowing down respiratory metabolism in strawberries. Moreover, the gradual release of TEO from the chitosan nanogel matrix likely disrupted enzyme activity, particularly those involved in cell wall softening, thereby reducing the rate of pectin hydrolysis and contributing to better firmness retention. These findings align with previous reports showing that coatings enriched with citrus essential oils reduced metabolic rates and preserved the firmness of strawberries by inhibiting enzymes responsible for pectin degradation [43]. Furthermore, a recent study by Li et al. [44] demonstrated that nanoencapsulation of thyme essential oil using chitosan cross-linked with sodium tripolyphosphate (TPP) preserved strawberry firmness over 7 days of storage at 25 °C. In contrast, this study used stearate ester as the cross-linking agent in chitosan–thyme oil nanogels and observed substantial firmness retention over an extended 16-day storage period at 4 °C.

    Total soluble solids (TSS), titratable acidity (TA), and pH

    The changes in TSS content in control and treated strawberry fruits during storage are presented in Fig. 6a. The most pronounced variation in TSS was observed in the control group, followed by the treatment with a higher concentration of free essential oil (E2). In these two treatments, TSS levels initially increased by day 4, followed by a sharp decline from day 8 until the end of the storage period, compared to the other treatments. The remaining treatments exhibited a similar trend, though the changes were less intense and progressed more gradually. Specifically, the highest TSS values in the NE2, NE1, and E1 treatments decreased from an initial 7.2 °Brix at the start of storage to 7.13, 6.53, and 6.56 °Brix, respectively, by the end of the storage period.

    The observed increase in TSS accumulation in fruits coated with nano-encapsulated thyme essential oil in chitosan nanogels may be attributed to a reduction in respiratory rate and overall metabolic activity, resulting in lower consumption of sugars through these pathways. Furthermore, interactions between the essential oil and cellular membranes may influence the fruit’s metabolic pathways and senescence processes. Moreover, interactions between the essential oil and cellular membranes may influence the fruit’s metabolic pathways and senescence process. The observed reduction in TSS content at higher concentrations of free essential oil indicates a potential adverse effect on fruit tissue, possibly accelerating ripening or senescence, consistent with previous reports [9]. Additionally, Velichkova et al. [45] reported that reduced weight loss may lead to an apparent increase in sugar concentration due to lower water loss, while decreased decay incidence can limit sugar consumption by fungal pathogens.

    According to Fig. 6b, the trend of changes in titratable acidity (TA) of strawberry fruit juice during storage showed a general decline from day 4 until the end of the storage period, except for the control fruits, which exhibited an increasing trend from day 12 to day 16. The most minor in TA was observed in the NE2 and NE1 treatments, where acidity levels decreased from an initial 5.8% to 5.09% and 4.32%, respectively, by day 16. In contrast, the TA in the control group decreased from 5.8% at the beginning to 3.6% by day 12, followed by an increase to 6.5% by the end of storage.

    As expected, with prolonged storage and the progressive loss of organic acids, the pH of the fruit juice exhibited an increasing trend (Fig. 6c). The evaluation of pH levels in both control and treated fruits (with free and nano-encapsulated thyme essential oil) revealed statistically significant differences among treatments. The highest pH value (4.25) was recorded in the control fruits after 16 days, while the corresponding values for the E1, E2, NE1, and NE2 treatments were 4.18, 4.23, 4.16, and 4.16, respectively. The most effective preservation of organic acid content and pH stability was achieved in fruits coated with nano-encapsulated thyme essential oil in the chitosan nanogel matrix.

    Fig. 6
    figure 6

    Effects of encapsulated thyme essential oil on TSS a TA b and, pH c of strawberry fruits during 16 days of storage at 4 °C. Data were represented as means ± standard deviation (n = 3). Treatments: C (control), E1 (thyme essential oil coating 600 µL/L), E2 (thyme essential oil coating 1200 µL/L), NE1 (encapsulated thyme essential oil coating 600 µL/L) and NE2 (encapsulated thyme essential oil coating 1200 µL/L)

    The reduction in TA and the concurrent increase in pH is commonly associated with enhanced respiratory activity and the progression of fruit senescence. These changes are primarily attributed to the consumption of organic acids during respiration [46]. It appears that the application of essential oil coatings—particularly nano-encapsulated thyme essential oil within a chitosan matrix—effectively reduced metabolic activity and respiration rate, likely by partially blocking stomata and limiting oxygen diffusion. These findings are consistent with previous reports on the use of CMC coatings combined with garlic oil, which extended the shelf life of strawberries by modulating physiological processes [9]. The gradual increase in titratable acidity observed at the end of the storage period in control fruits may be attributed to a potential rise in fungal population. Increased fungal activity can lead to the production of acidic metabolites and mucilage in the fruit tissue—a phenomenon also reported in earlier studies involving edible coatings based on chitosan and Aloe vera extract applied to blueberries [47].

    Total anthocyanin content

    The changes in total anthocyanin content of strawberry fruits during storage revealed significant differences over time (Fig. 7a). The highest accumulation of anthocyanins was observed in the untreated control fruits and those treated with a high concentration of free essential oil (E2) by the end of the storage period. In contrast, the initial anthocyanin content was 18.34 mg/kg. The lowest increase and the least fluctuation in anthocyanin levels were recorded in fruits treated with nonencapsulated thyme essential oil in chitosan nanogels, followed by treatment E1. The percentage increases in anthocyanin content compared to day 0 for the control, E1, E2, NE1, and NE2 treatments were 55%, 30%, 39%, 31%, and 17.7%, respectively.

    The significant rise in anthocyanin, particularly in control fruits, may reflect more advanced ripening during storage and greater weight loss, both of which can influence pigment concentration. However, strawberries treated with thyme essential oil showed lower anthocyanin levels than the untreated fruits. Moreover, statistically significant differences in anthocyanin content were observed among fruits coated with nano-encapsulated thyme essential oil. Anthocyanins, the red pigments in strawberries, are classified as polyphenols. Their biosynthesis is regulated by the activity of phenylalanine ammonia-lyase (PAL) [48]. Key factors influencing total phenolic and anthocyanin synthesis include cultivar and fruit maturity at harvest. The reduced accumulation of anthocyanins in coated fruits could be due to delayed biosynthesis of anthocyanins and other red pigments, consistent with findings from previous studies [49]. Similar results have been reported for other fruits treated with chitosan-based coatings [50]. Notably, more favorable results were achieved when thyme essential oil was nano-encapsulated in chitosan nanogels, effectively mitigating the potential tissue-damaging effects of high concentrations of free essential oil. This encapsulation provided a more controlled release and prolonged effectiveness, as observed in the NE1 and NE2 treatments. These findings align with prior research on the application of lemongrass essential oil in alginate-based edible coatings to extend shelf life and preserve quality in fresh-cut pineapple [51]. The reduced anthocyanin accumulation in treated fruits may also be linked to the semi-permeable properties of thyme essential oil, which can reduce oxygen diffusion and increase localized CO₂ concentration around the fruit surface. This shift in gas composition could suppress biochemical pathways responsible for anthocyanin biosynthesis [52]. Similarly, Pelayo et al. [53] reported that elevated CO₂ levels during storage inhibited anthocyanin synthesis in strawberries. In addition, essential oil treatments can significantly reduce fruit respiration rates, thereby minimizing water loss from the surface, which may explain the lower anthocyanin concentrations in coated fruits compared to uncoated controls. Thus, treatment with essential oils can effectively delay fruit ripening and enhance the postharvest shelf life of strawberries.

    Ascorbic acid

    One of the key factors in preserving the quality attributes of fruits during storage is their ascorbic acid content [54]. Figure 7b shows the changes in ascorbic acid concentration in strawberries during storage. Total ascorbic acid levels in both control and treated fruits decreased significantly over time. At the end of the storage period, strawberries treated with nano-encapsulated thyme essential oil exhibited higher ascorbic acid levels compared to the control and to those coated with the non-encapsulated essential oil. The most significant reduction in L-ascorbic acid content was observed in the control fruit, decreasing from 52.6 to 17.74 mg/100 g FW, whereas the smallest decrease was seen in treatment NE2, with a final value of 25.98 mg/100 g FW. This decline may be attributed to enhanced oxidation caused by the accumulation of reactive oxygen species, such as superoxide and hydroxyl radicals, in strawberries [14]. The present findings indicate that nanoencapsulation of thyme essential oil in a chitosan-based nanogel can more effectively preserve vitamin C during storage compared to the free essential oil. This may be attributed to the antioxidant properties of the essential oil, as illustrated in Fig. 7b, as well as its ability to reduce oxygen diffusion and consequently lower the respiration rate in treated fruits, thereby slowing the oxidation process of ascorbic acid [55, 56]. These results are consistent with previous studies reporting that edible coatings containing citrus essential oil improved postharvest attributes of strawberries more effectively than lemon oil coatings [43].

    Fig. 7
    figure 7

    Effects of encapsulated thyme essential oil on the anthocyanin content a and, ascorbic acid b of strawberry fruits during 16 days of storage at 4 °C. Data were represented as means ± standard deviation (n = 3). Treatments: C (control), E1 (thyme essential oil coating 600 µL/L), E2 (thyme essential oil coating 1200 µL/L), NE1 (encapsulated thyme essential oil coating 600 µL/L) and NE2 (encapsulated thyme essential oil coating 1200 µL/L)

    Polyphenol oxidase (PPO) activity

    During storage, PPO activity in the control fruits increased sharply. In contrast, this increase was significantly (P ≤ 0.05) suppressed in coated samples, particularly in treatments with nanochitosan-encapsulated thyme essential oil (Fig. 8). At the end of the storage period, the lowest PPO activity was observed in NE2 and NE1 treatments, followed by E1. PPO catalyzes the oxidation of phenolic compounds to o-quinones in the presence of oxygen, which are responsible for enzymatic browning in fresh produce tissues. This enzyme can be considered a key indicator for assessing senescence during the postharvest period in fresh fruits and vegetables [57]. The present findings indicate that thyme essential oil, when applied with enhanced stability in the NE2 coating, inhibited oxygen penetration into fruit tissues and prevented tissue degradation, thereby reducing PPO activity. These findings align with previous studies showing that chitosan coatings containing thymol (0.02%) or geraniol (0.04%) effectively reduce PPO activity and extend strawberry shelf life up to 7 days at 4 °C [50]. In the present study, nano-chitosan encapsulation of thyme essential oil appears to further prolong strawberry shelf life compared to these earlier reports.

    Fig. 8
    figure 8

    Effects of encapsulated thyme essential oil on the PPO activity of strawberry fruits during 16 days of storage at 4 °C. Data were represented as means ± standard deviation (n = 3). Treatments: C (control), E1 (thyme essential oil coating 600 µL/L), E2 (thyme essential oil coating 1200 µL/L), NE1 (encapsulated thyme essential oil coating 600 µL/L) and NE2 (encapsulated thyme essential oil coating 1200 µL/L)

    Antioxidant enzymes

    The activities of the antioxidant enzymes peroxidase (POD) and catalase (CAT) increased markedly in coated strawberries compared with the control fruits throughout the storage period (Fig. 9a and b). In the control group, POD and CAT activities began to decline on days 12 and 8, respectively. In treatments containing free essential oil, the initial increase in enzyme activity was slightly higher than that.

    Fig. 9
    figure 9

    Effects of encapsulated thyme essential oil POD a and, CAT b activity of strawberry fruits during 16 days of storage at 4 °C. Data were represented as means ± standard deviation (n = 3). Treatments: C (control), E1 (thyme essential oil coating 600 µL/L), E2 (thyme essential oil coating 1200 µL/L), NE1 (encapsulated thyme essential oil coating 600 µL/L) and NE2 (encapsulated thyme essential oil coating 1200 µL/L)

    in the nanochitosan-encapsulated essential oil treatments until the mid-storage period; however, the latter showed a significantly greater increase thereafter until the end of storage. By the end of storage, POD and CAT activities in the control were recorded at 3.0 and 0.6 U/mg FW, respectively, whereas the highest values were observed in NE2, reaching 9.02 and 6.7 U/mg FW, respectively.

    The activation of plant antioxidant defense systems—both enzymatic and non-enzymatic—plays a crucial role in mitigating oxidative damage caused by the accumulation of reactive oxygen species (ROS) [16]. The sustained and pronounced increase in POD and CAT activities observed in strawberries treated with nanochitosan-encapsulated thyme essential oil compared to the control and other treatments clearly indicates suppression of ROS in the treated fruits. Although free thyme essential oil also reduced ROS levels more effectively than the control, its protective effect diminished over time.

    POD and CAT mitigate oxidative stress by converting hydrogen peroxide (H₂O₂) into water and oxygen, thereby preventing its deleterious effects [58]. In the present study, the prevention of cellular degradation in strawberries may be attributed to the enhanced POD and CAT activities induced by the encapsulated essential oil, facilitating the breakdown of H₂O₂ into harmless products. In agreement with our findings, Adiletta et al. [59] reported that chitosan/nano-silica coatings increased CAT and SOD activities in loquat fruits, while Hassan et al. [16] demonstrated that thyme essential oil encapsulated in chitosan enhanced CAT and SOD activities in sweet basil leaves, thereby extending their shelf life. It appears that the elevated antioxidant enzyme activities in NE2-treated strawberries contributed to reducing H₂O₂-induced cellular damage and preventing tissue degradation, ultimately prolonging their storage life.

    Antioxidant activity

    Throughout the storage period, antioxidant activity in all treatments and the control increased until day 8, followed by a gradual decline until the end of storage (Fig. 10). Strawberries coated with chitosan-based formulations exhibited the most minor fluctuations compared with other treatments. In NE2, antioxidant activity increased from 20.1% to 26.16%, whereas the lowest value was recorded in the control fruit at 10.73%. The highest antioxidant activity was observed in coatings containing nanochitosan-encapsulated thyme essential oil at concentrations of 1200 and 600 µL/L (NE2 and NE1), followed by the free essential oil coating (E1).

    Fig. 10
    figure 10

    Effects of encapsulated thyme essential oil on the antioxidant activity of strawberry fruits during 16 days of storage at 4 °C. Data were represented as means ± standard deviation (n = 3). Treatments: C (control), E1 (thyme essential oil coating 600 µL/L), E2 (thyme essential oil coating 1200 µL/L), NE1 (encapsulated thyme essential oil coating 600 µL/L) and NE2 (encapsulated thyme essential oil coating 1200 µL/L)

    The decline in antioxidant activity in the control fruit at the end of storage can be attributed to natural senescence and spoilage. These results suggest that encapsulation of thyme essential oil in nanochitosan not only extends shelf life but also preserves higher antioxidant activity in strawberries over prolonged storage. Thymol, one of the most common phenolic compounds in essential oils, has been previously reported to possess potent antioxidant properties [50, 60]. Consistent with our findings, Wang and Gao [29] reported that chitosan coatings enhanced radical scavenging capacity, phenylpropanoid compound levels, and antioxidant enzyme activities in strawberries. Moreover, the antioxidant activity of chitosan can be further enhanced by incorporating compounds such as certain sugars, essential oils, and gelatin [61]. Our findings are also in agreement with earlier reports indicating that the antioxidant mechanism of chitosan may arise from its ability to chelate metal ions such as copper and iron at enzyme active sites, thereby inactivating oxidizing enzymes, and/or to interact with lipids in these enzymes [50, 60].

    Pathogenic contamination assessment

    Microbial colony enumeration is an effective method to evaluate the antimicrobial performance of different coating formulations for strawberry preservation. According to established standards, fresh fruits or vegetables are considered unsuitable for consumption when total microbial counts exceed 4 Log CFU/g [62]. As shown in Fig. 11, a gradual increase in microbial colony numbers over time was evident across all treatments.

    Fig. 11
    figure 11

    Effects of encapsulated thyme essential oil on the decay incidence of strawberry fruits during 16 days of storage at 4 °C. Data were represented as means ± the standard deviation (n = 3). Treatments: C (control), E1 (thyme essential oil coating 600 µL/L), E2 (thyme essential oil coating 1200 µL/L), NE1 (encapsulated thyme essential oil coating 600 µL/L) and NE2 (encapsulated thyme essential oil coating 1200 µL/L)

    Among the groups, untreated fruits (control) exhibited the highest microbial proliferation. At the beginning of storage, microbial counts in the control group were 0.85 Log CFU/g, which rose to 4.75 Log CFU/g by day 8—exceeding the permissible consumption threshold. By day 16, colony counts in the control group were 1.5-fold higher than those on day 8, indicating severe microbial contamination. In contrast, samples treated with NE2 showed a markedly slower increase in colony numbers. However, counts rose during storage, the final value was only two logarithmic cycles higher than the initial level and remained below the unfit-for-consumption limit. This suggests that NE2 treatment significantly suppressed microbial growth up to day 16. Under cold storage conditions, the shelf life of strawberries varied between 4 and 16 days depending on the treatment. The application of free thyme essential oil in coatings also delayed microbial growth, extending fruit longevity by approximately four and eight days in treatments E1 and E2, respectively.

    Since fungal spoilage is one of the primary factors reducing strawberry quality and shelf life, a separate evaluation of yeast and mold growth across treatments was conducted [63]. As depicted in Fig. 11, yeast and mold counts increased progressively in all groups during storage. However, coatings containing chitosan-encapsulated thyme essential oil exhibited slower fungal proliferation, particularly in NE2, where the essential oil concentration was doubled. In this treatment, yeast and mold counts were reduced from 7.41 to 3.10 Log CFU/g compared with the control, representing a decrease of more than two logarithmic cycles. These findings are consistent with the results of De Oliveira Filho et al. [64] and Li et al. [44], who also reported that nano-chitosan coatings containing thyme essential oil and nano-emulsions with Cymbopogon martinii or Mentha spicata essential oils significantly inhibited yeast and mold growth compared to essential-oil-free controls [44, 64]. This antimicrobial effect is likely due to the combined action of chitosan and thyme essential oil, which disrupts fungal membrane integrity, thereby reducing contamination and inhibiting respiration [65].

    Overall, these results indicate that nanochitosan-encapsulated thyme essential oil at a concentration of 1200 µL/L plays a critical role in reducing fungal growth and extending strawberry shelf life. The NE2 coating formulation effectively prevented yeast and mold proliferation, thereby mitigating fruit spoilage during storage.

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  • Development of a patient-specific epicardial guide for ventricular tachycardia ablation surgery using high-consistency rubber silicone molding | BioMedical Engineering OnLine

    Development of a patient-specific epicardial guide for ventricular tachycardia ablation surgery using high-consistency rubber silicone molding | BioMedical Engineering OnLine

    This preliminary study is the first to assess the use of HCR wrapping for patient-specific guides in VT scar ablation.

    Innovative solutions are needed to precisely identify and target complex epicardial substrate during concomitant open-chest surgery—enabling accurate localization on the arrested heart—and in stand-alone procedures, where prolonged surgery and the need for an EP team in the room present additional challenges. Epicardial guides would allow the surgeon to improve navigation, accuracy, safety, and therapeutic efficacy in difficult-to-treat ablation patients.

    Herein, we aimed to develop a feasible, cost-effective, and rapid method for creating epicardial surgical guides, focusing on early feasibility and technical viability. Compared to traditional 3D printing methods, HCR molding offers an efficient and affordable alternative that yields promising potential.

    Rationale behind silicone and molding

    The emergence of 3D printing technologies in the last decade has created abundant application opportunities in the field of precision medicine and surgical education [10]. However, 3D printing resins face challenges, such as brittleness, poor dimensional accuracy and low mechanical strength, especially after post-processing and sterilization, as well as limited material options, high costs, and lack of standardization [11,12,13]. In addition, uncured monomers or leachates may release cytotoxic byproducts, further complicating their use in surgical environments [14]. To overcome the limitations of traditional 3D printing resins, we chose a silicone-based solution—an elastomer widely used in the medical field [15]. Some silicones are approved for long-term implantation and have a proven track record in dynamic (cardiovascular) surgical environments. With its flexibility, durability, stability, versatility, and resistance to extreme temperatures, silicone can be fully customized to meet the specific needs of this epicardial application.

    Although silicone modeling has been used to create customized molds [16,17,18], no studies have fabricated silicone specifically as an epicardial guide, posing unique challenges in identifying a material that meets all necessary criteria. Material choice is a critical factor to the success of a patient-specific medical device. Properties such as silicone hardness and flexibility help the guide maintain shape and function despite the heart’s constant motion, which prevents gaps that could lead to uneven lesions.

    We also considered liquid silicone rubber molding, a widely used technique in medical device manufacturing and studies. However, this method requires creating an inverted mold with several components (multi-part mold system, vent channels, and silicone channels) that must be precisely aligned and securely connected using interlocks to prevent movement. This makes the process significantly more labor-intensive and less suitable for a streamlined, efficient, and rapid customized workflow for ablation treatments. Another barrier is that the liquid silicone for our required durometer would be too viscous at room temperature, making injection into the mold difficult and prone to air entrapment. We also considered direct 3D silicone printing, a more novel approach. However, it was not our preferred choice due to potential challenges with support structures, layer adhesion, and viscosity control, which could compromise integrity and make the process tedious.

    An advantage of silicone wrapping is that sculpting results in a smoother inside surface due to the natural smoothness of milled silicone; unlike 3D printing, which can create rougher surfaces. This technique also allows customization using different silicones, curing times, and pigments to meet tailored requirements.

    Thermal insulation and procedural safety

    The guide manufacturer is responsible for ensuring compliance with design requirements (form, fit, and function) throughout the entire device’s life cycle [19]. A critical consideration in the development of surgical guides is their thermal insulation performance under ablative stress. These properties are vital for two primary reasons: ensuring procedural safety during targeted ablation and protecting adjacent, non-target anatomy from thermal injury. The material’s electrical non-conductivity reduces the risk of unintended heating when ablation instruments come into accidental contact with the guide’s edges—a scenario that can result in localized temperatures as high as 70–90 °C during radiofrequency ablation or below 0 °C during cryo-ablation, potentially causing collateral tissue damage. Due to the lack of conductivity data in the material datasheet, preliminary tests were conducted simulating accidental instrument-guide contact. These experiments confirmed that the guide effectively prevented thermal injury to underlying tissue. However, further comprehensive thermal and mechanical characterization—particularly post-processing and sterilization—remains necessary to validate the material’s suitability for clinical application.

    Design considerations

    Guide stability on the epicardial surface is of critical importance to ensure precise lesion placement. Material testing indicated that silicone with a 70 Shore A hardness and 2–3 mm thickness provided an optimal balance between flexibility and intraoperative stability on the beating heart. In addition, preliminary foldability tests were conducted to assess suitability for minimally invasive approaches, revealing that the current guide designs are too bulky for efficient trocar introduction, largely due to excess material at the apex base. However, this limitation is expected to be less significant in smaller, target-specific guide designs. Future work should explore alternative silicones and ensure uniform thickness without excess material. Moreover, to facilitate insertion and removal of the guide through a trocar, incorporating a secure locking system with small tabs on the lateral sides that click when folded might prove useful.

    Finally, an important challenge in epicardial guide use for minimally invasive stand-alone ablation is achieving stable anchoring of the model on a beating heart. Potential solutions we considered include micro-suction ports, adhesive surfaces, or specialized gripping materials.

    Broader clinical applicability and translational pathway

    While the guide in this feasibility study was designed to cover a broad scar region, the end objective is to use preprocedural input from the EP to incorporate predefined ablation targets in form of ‘windows’ in the mask, directly providing the surgeon with an ablation roadmap or lesion set. By providing intra-operative guidance, it could enhance precision and reduce reliance on full real-time mapping and a complete EP team, since lesions are identified pre-operatively. However, the customization of these ablation openings requires further finetuning as they depend on patient pathology, surgical approach (open-chest or minimally invasive), and the type and size of the catheter used.

    A crucial early step in clinical translation is assessing biocompatibility and characterizing the sterilized silicone material to confirm its suitability, thermal stability, mechanical integrity, and overall safety for clinical use. Preclinical validation is essential to rule out risks, such as loose fragments or allergic reactions, and to assess the guide’s fit, stability, efficacy and precise electro-anatomic alignment. Swine models are commonly used due to physiological similarity to humans and comparable heart rates, beginning with open-chest studies and advancing to minimally invasive approaches [20]. For these experiments, subject-specific guides are fabricated from pre-operative cardiac CT. While device alignment partially relies on visible anatomical landmarks (aorta, pulmonary artery ring, and left ventricular apex)—which are accessible via both open-chest and thoracoscopic approaches—epicardial electro-anatomic mapping is still needed during early translational phases. Alignment accuracy can be quantified by measuring the percentage overlap between pre-operative targets (scar and mapping) and post-ablation areas confirmed by mapping, and post-explant TTC staining and histology.

    The expected benefits—allowing concomitant VT ablation, improved consistency, shorter procedures, fewer repeat ablations—translate into both clinical and economic value. Though regulatory and compliance efforts for clinical translation are significant, they are minor compared to the costs of repeat ablations, prolonged procedures, and hospital stay/visits—especially once integrated into routine practice. While the training and integration of the device into the surgical workflow may initially add time to the procedure, it is expected to reduce overall operative time over the long term—particularly if intraoperative mapping can be reduced, potentially eliminating the need for an EP team in the operating room.

    This silicone-based approach can serve as a basis for the engineering and development of other customizable guides. The potential for further applications is vast, as this can be adapted to meet specific mapping and procedural requirements. For example, our group previously developed an epicardial guide for Brugada syndrome ablation, highlighting the arrhythmogenic substrate in the right ventricular outflow tract, based on electro-anatomical mapping data [21]. In addition, we have created guides for coronary artery mapping and optimal bypass target placement during coronary artery bypass grafting [22], and sinus node protection for inappropriate sinus tachycardia ablation. Furthermore, this approach could extend to other (non-)cardiac procedures for anatomical modeling, pre-operative planning, and surgical training.

    Limitations and future considerations

    Several limitations should be considered. As this was an early proof-of-concept study, the 3D-printed mold was fabricated from non-biocompatible PLA. Thus, biocompatibility testing, including in vitro cytotoxicity assessments as per ISO 10993–5:2009 (https://www.iso.org/standard/36406.html), as well as post-sterilization material characterization assessments are warranted for future research. Following, in vivo porcine experiments are needed on the beating heart to assess stability, fit, ablation efficacy, and electro-anatomic alignment more in-depth.

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  • TGF-β signaling and tumor microenvironment dynamics in bladder cancer progression post-BCG therapy: a longitudinal single-nucleus RNA-seq study | BMC Cancer

    TGF-β signaling and tumor microenvironment dynamics in bladder cancer progression post-BCG therapy: a longitudinal single-nucleus RNA-seq study | BMC Cancer

    Landscape of single cell nuclei of bladder cancers

    Nine patients diagnosed with NMIBC were enrolled in this study. All patients were treated with adjuvant BCG after endoscopic resection. Three patients experienced disease progression. They were classified as “progressors”. The remaining six patients who did not experience disease progression were classified as “non-progressors.” Detailed clinicopathological information of these patients is provided in Fig. 1A.

    To characterize cell types in the tumor microenvironment (TME) of NMIBC, a total of 14 specimens (P-2, P-8 and P-9 specimens per patients including three post-progression specimens, 1 cell-free specimen) were subjected to snRNA-seq using resection specimens before BCG treatment. A total of 73,979 nuclei were obtained after removing cell doublets (median of 5,804 nuclei, 1,000–8,720 nuclei per patient) and profiled for mRNA quantification. Detailed information on snRNA-seq is available in Supplementary Table S1. Clustering of snRNA-seq revealed six major cell types: 61,837 epithelial cells, 898 endothelial cells, 5,973 T cells, 1,907 B cells, 1,712 myeloid cells, and 1,652 fibroblasts.

    A t-distributed stochastic neighbor embedding (tSNE) plot was used to illustrate cell distribution as shown in Fig. 1B. Notably, epithelial cells constituted a significant fraction of cells, indicative of high tumor purity across samples. Moreover, heterogeneity between Progression and NP samples was observed as shown in pairwise tSNE plots (P-2, P-8, and P-9), Primary and progression samples formed well-separated clusters, indicating distinct transcriptional differences (Fig. 1C). The expression of marker genes for each major cell type is depicted in Fig. 1D, including CDH1 [18] for epithelial cells, VWF [19] for endothelial cells, COL1A1 [20] for fibroblasts, CSF1R [21] for myeloid cells, CD2 [22] for T cells, and MS4A1 [18] for B cells. Additionally, cluster-specific gene expression of selected markers is presented in Fig. 1E.

    The TGF-β signaling pathway is activated in malignant cells with disease progression

    Epithelial cells were further subject to fine-clustering. tSNE plots of epithelial cells revealed that the clustering of epithelial cells was largely determined with respect to individual patients, indicative of a substantial level of inter-patient heterogeneity of malignant cells (Fig. 2A). We then performed inferCNV to determine copy number alterations (CNA) based on gene expression. Using inferCNV analysis, we identified significant interpatient heterogeneity in inferred CNA profiles across nine bladder cancer malignant cells (Fig. 2B). Notable arm-level gains were observed in chromosomes 1q, 5p, and 20p, while frequent losses were detected in 9p, 11p, 17p, and 19p. Hurst et al. [23] have reported that gains of 1q and 5q and losses of 9p, 11p, 17p, and 19p are associated with disease progression of bladder cancers. In addition, Bellmunt et al. [24] have found that gains in 20p are linked to disease recurrence, while López et al. [25] have identified a connection between gains in 5q and bladder cancer recurrence.

    Fig. 2

    Identification of malignant and defined malignant functions using ssGSEA (A) tSNE plot of 61,837 epithelial cells. (B) Chromosomal gains and losses in malignant epithelial cells were predicted using inferCNV. (C) A heatmap displaying enrichment scores from the ssGSEA analysis and correlation values between the ssGSEA score and monocle pseudotime results of the progression samples by Cell ID. (D)-(F) Monocle analysis of the development of malignant cells between P-2/P-8/P-9 Progression (Before BCG) and P-2/P-8/P-9 (Progression) with pseudotime, as well as the pseudotime trajectory of TGF-β signaling for P-2/P-8/P-9 Progression (Before BCG) and P-2/P-8/P-9 (Progression)

    We analyzed malignant cells using longitudinal specimens obtained before treatment and after disease progression. Functional gene sets were identified using ssGSEA with representing sample-specific disease progression using Hallmark pathway. This analysis identified molecular functions enriched in the difference of NES values between Progression and Primary specimens. It highlighted tumor progression-related functions, such as WNT–β-catenin signaling and TGF-β signaling, which are consistently enriched across malignant cells during disease progression (Fig. 2C). This result suggests that TGF-β might play a key progression-related molecular function in disease progression after BCG (Figs. 2D–2F). Pseudo-temporal analysis representing the transition from treatment-naïve to progression samples indicated that each paired sample of malignant cells passed through smooth one-dimensional curves. Specifically, cells from treatment-naïve samples were ordered at early pseudotime points, followed by cells from progression samples at later pseudotime points. This indicates a clear separation in differentiation trajectories of treatment-naïve and progression cells. Results of the ssGSEA analysis of TGF-β signaling along the pseudotime trajectory showed that TGF-β signaling enrichment scores increased progressively from treatment-naïve to progression samples, consistent with the trajectory analysis.

    TME cell subtypes associated with disease progression

    Sub-clustering of macrophages (Fig. 3A; proliferating, M2 and SPP1 + macrophages) revealed three distinct subtypes. Pseudotime trajectory analysis showed bifurcation into two major branches, with proliferating and some subtypes located in earlier stages, M2 macrophages diverging into another branch, and SPP1 + macrophages appearing in the later phase (Fig. 3B). M2 macrophages were relatively more abundant in the Progression group than in the Progression-Before BCG group. Proliferating myeloid cells showed a gradual increase from NP to Progression-Before BCG and remained elevated in the Progression group, whereas SPP1 + macrophages exhibited a consistent decrease across groups, with the lowest proportion observed in the Progression group (Fig. 3C). The top 10 marker genes of macrophage subtypes are shown in Fig. 3D. The LYZ gene was identified as a marker gene for proliferating myeloid cells [26, 27]. In previous studies on bladder cancer, LYZ has been used as a myeloid cell marker. Moreover, Gu et al. have demonstrated a significant association between these cells and proliferation. Regarding the TTC7B gene, He et al. have found notable correlations between its expression and infiltration levels of various immune cells, including M2 macrophages [28].

    Fig. 3
    figure 3

    Characterization and profiling of four subtypes. A tSNE plot of 508 macrophage cells, showing the composition of three main subtypes. B Monocle analysis of the development between subtypes with pseudotime. C Cell proportion of subtypes by NP, Progression Before BCG, and Progression. D Heatmap of Top 10 marker genes of each subtype. E tSNE plot of 245 dendritic cells, showing the composition of 3 main subtypes. F Monocle analysis of the development between subtypes with pseudotime. G Cell proportion of subtypes by NP, Progression Before BCG, and Progression. H Heatmap of Top 10 marker genes of each subtype. I tSNE plot of 1,652 fibroblasts, showing the composition of the 3 main subtypes. (J) Monocle analysis of the development between subtypes with pseudotime. K Cell proportion of subtypes by NP, Progression Before BCG, and Progression. L Heatmap of Top 10 marker genes of each subtype. M tSNE plot of 898 endothelial cells, showing compositions of 4 main subtypes. N Monocle analysis of development between subtypes with pseudotime. O Cell proportion of subtypes by NP, Progression Before BCG, and Progression. P Heatmap of Top 10 marker genes of each subtype

    Sub-clustering of dendritic cells (Fig. 3E; DC1, DC2, and DC3) demonstrated sequential differentiation. The pseudotime trajectory (Fig. 3F) followed an order from DC3 (LAMP3 +) at the earliest stage, through DC2 (CD1A +), and finally to DC1 (XCR1 +) at the latest stage, highlighting dynamic changes in cell states. For dendritic subtypes, CD1A (DC2) cells showed only a modest and variable increase in the Progression group compared with NP and Progression-Before BCG, without a consistent trend across patients (Fig. 3G). Dendritic cells are known for their roles in antigen presentation and immune response orchestration. They exhibited CADM1 expression, particularly in XCR1 (DC1), as highlighted by our DEG results where CADM1 emerged as a marker (Fig. 3H). To understand CADM1’s function [29], future research needs to be conducted on tumor progression where CADM1’s role has been extensively studied. Kang et al. have found that the ADGRG5 gene in CD1A (DC2) is associated with dendritic cells, showing a stronger correlation with ADGRG5 expression than in other types of immune cells tested [30]. In our study, ADGRG5 was identified as a marker in DC2. In DCs, CCR7-dependent migration from peripheral tissues to lymphoid tissues plays a critical role in host defense against pathogens and immune tolerance maintenance [31]. This migratory process is governed by complex intracellular signaling pathways that can regulate both DC movement and inflammatory responses. Our analysis identified CCR7 as a marker gene specifically in LAMP3-expressing DCs (DC3), as shown in our differential gene expression data (Fig. 3H).

    Fibroblast subtypes (Fig. 3I; myoCAF, iCAF, and SMC) also showed distinct developmental patterns. Pseudotime analysis (Fig. 3J) indicated that myoCAF and iCAF dominated the early phase, while SMC and iCAF became predominant later, with the trajectory culminating in iCAF and SMC states. Results revealed a higher proportion of myoCAF cells in the Progression Before BCG group compared with the Progression group. Additionally, SMC cells showed a consistent proportion across all groups. Notably, iCAF cells maintained the highest proportion in all groups, although they were slightly reduced in the Progression group compared with NP and Progression Before BCG groups (Fig. 3K). Both iCAF and myoCAF showed similar cell proportions, although iCAF demonstrated a stronger correlation with reduced survival rates. The top 10 marker genes of fibroblast subtypes are displayed in Fig. 3L.

    Sub-clustering of endothelial cells (Fig. 3M; venous, tip-like, arterial, and lymphatic) revealed further heterogeneity. Pseudotime trajectory analysis (Fig. 3N) indicated that venous and tip-like cells appeared in early stages, arterial and lymphatic cells arose but later disappeared, leaving tip-like cells predominant at the end. The distribution of endothelial cell subtypes within each group is illustrated in Fig. 3O. In the NP group, venous cells constituted the highest proportion, followed by tip-like cells. Arterial and lymphatic cells were present in relatively smaller proportions. In the Progression Before BCG group, tip-like cells dominated almost entirely, with a small presence of venous cells. Arterial and lymphatic cells were nearly absent. In the Progression group, tip-like cells also constituted the highest proportion, although the proportion of venous cells was significantly reduced compared with levels in NP and Progression-Before BCG groups. Additionally, proportions of arterial and lymphatic cells slightly increased. The top 10 marker genes of endothelial subtypes are shown in Fig. 3P. In humans, loss-of-function mutations in VEGFC or VEGFR3 can lead to lymphedema, while the application of recombinant VEGFC can stimulate robust lymphangiogenesis in adults, suggesting its therapeutic potential for lymphedema and tissue repair.

    To distinguish the function of each subtype, we performed hallmark pathway analysis using ssGSEA and selected the top 10 hallmark pathways. As shown in Supplementary Fig. 1A, Proliferating Macrophage subtype was enriched in the EMT and MYC-TARGETS-V2 pathways, while the M2 subtype was enriched in the PI3K-AKT-mTOR-SIGNALING and TGF-BETA-SIGNALING pathways. Additionally, the SPP1 subtype was enriched in the MYC-TARGETS-V1, HYPOXIA, and DNA-REPAIR pathways. The population of dendritic cells was clustered into three subtypes (DC1, DC2, and DC3), although they were not clearly separated in ssGSEA Hallmark pathway. However, DC2 tended to be enriched in the DNA-REPAIR pathway, while DC showed a tendency to be enriched in the EMT pathway (Supplementary Fig. 1B). Additionally, fibroblasts were divided into three sub-clusters (myoCAF, iCAF, and SMC). The myoCAF subtype was enriched in immune-related pathways and the INFLAMMATORY-RESPONSE pathway, whereas iCAF and SMC subtypes were not clearly distinguished as shown in Supplemantary Fig. 1C. The venous subtype, a subset of endothelial cells, was enriched in immune-related pathways and the INFLAMMATORY-RESPONSE pathway, while the Tip-like sub-cluster was enriched in the MYC-TARGETS-V1 pathway. Additionally, the arterial subtype was enriched in the COMPLEMENT and ANGIOGENESIS pathways, whereas the Lymphatic sub-cluster was enriched in the NOTCH-SIGNALING pathway, as shown in Supplementary Fig. 1D.

    Cell–cell interactions across malignant cells and TME cells

    To investigate differences in cell–cell communication among various cell subpopulations, we analyzed interaction numbers and interaction strength using CellChat [16]. Our study included three conditions: No Progression (NP), Primary Before BCG, and Progression. Results in Fig. 4A illustrate variations in interaction patterns across these conditions. In the NP condition, interactions were relatively sparse, with notable communication occurring primarily between epithelial cells and other cell types. In the Primary condition (Before BCG), the interaction network became more complex, indicating increased communication among T cells, B cells, myeloid cells, and other subpopulations. The Progression condition showed the most extensive and robust interactions, suggesting a heightened level of cell–cell communication. We conducted a pathway-based ligand-receptor interaction analysis between TME cells. Results (Fig. 4A) demonstrated dynamic changes in cell–cell communication networks across different conditions, emphasizing the importance of epithelial cell interactions in NP, Progression Before BCG, and Progression. Notably, the interaction strength between epithelial cells and myeloid cells, as well as fibroblasts, increased from Progression Before BCG to Progression.

    Fig. 4
    figure 4

    Estimation of cell–cell communication and interactions between malignant and macrophage/fibroblasts using TGF- β signaling pathway geneset. A Interaction plot for major cell types. The strength of communication was assessed via CellChat across NP, Progression Before BCG, and Progression. B Using the TGF- β signaling pathway geneset in Nichenet, interactions between malignant cells and Macrophages were examined. C A heatmap displays active ligand-receptor pairs, highlighting interaction potential between receptors on malignant receiver cells and ligands on Macrophage sender cells. D Top 30 ligand-receptor (L-R) pairs ordered by interaction potential score. E Interactions between malignant cells and fibroblast using the TGF- β signaling pathway geneset in Nichenet. F A heatmap displays active ligand-receptor pairs, highlighting interaction potential between receptors on malignant receiver cells and ligands on fibroblast sender cells. G Top 30 ligand-receptor (LR) pairs ordered by interaction potential score

    According to Browaeys et al. [17], a validation study can substantiate the proposed methodology, illustrating that the final prior model of ligand–target interactions can be broadly applied to various biological systems, thereby supporting the applicability of Nichenet to a wide range of biological contexts. Consequently, we employed the TGF-β signaling pathway identified in Fig. 2 to analyze Nichenet. We defined the gene set of interest as those genes within the receiver cell type that are likely to be influenced by the cell–cell communication event.

    Results shown in Fig. 4A revealed that the interaction strength between epithelial cells and myeloid cells, as well as fibroblasts, increased from the Progression Before BCG stage to the Progression stage. Consequently, we analyzed ligand-receptor interactions during muscle regeneration in Progression samples using NicheNet. By applying stringent cutoffs, we identified differentially expressed ligands predicted to interact with receptors on malignant cell (top 20 by differential expression). We compiled a list of ligand-receptor pairs expressed in relevant cell types and predicted them to act upstream of TGF-b signaling specific genes. To narrow down the extensive list of candidates, we focused on ligands with high activation scores for TGF-b signaling genes and complementary targets in macrophages (Figs. 4B–4C) and fibroblasts (Figs. 4E–4F). In addition, we identified the top 30 ligand-receptor pairs in macrophages (Fig. 4D) and fibroblasts (Fig. 4G).

    Establishment and validation of prognostic significance marker genes

    Using TCGA data, we employed CIBERSORTx to estimate proportions of M2 macrophages and fibroblasts. As shown in Figs. 4D and 4G, top 30 ligand-receptor pairs were identified the. Based on these results, we examined the expression of the DSC2(L)-DSG2(R) pair in epithelial and myeloid cells and the expression of the ENG(L)-BMPR2(R) pair in epithelial and fibroblast cells from our snRNA-Seq data. We then analyzed their survival associations in TCGA BLCA data. Results are shown in Fig. 5. Patients with a higher average proportion of M2 macrophages and elevated expression of the DSC2(L)-DSG2(R) pair demonstrated a significant difference in survival with a p-value of 0.016 (Fig. 5A). Similarly, patients with a higher average proportion of fibroblasts and increased expression of the ENG(L)-BMPR2(R) pair also showed a significant survival difference, with a p-value of 0.017 (Fig. 5B). According to Nakauma-González et al. [32], DSC2 and DSG2 in bladder cancer are markers of intratumoral genomic and immunologic heterogeneity, particularly evident through squamous differentiation. This morphological heterogeneity can serve as a biomarker for intrinsic immunotherapy resistance in bladder cancer patients.

    Fig. 5
    figure 5

    Kaplan–Meier analysis of BLCA progression-free survival (PFS) associated with ligand–receptor pairs. A Expression heatmap and correlation matrix of DSC2 (myeloid cells) and DSG2 (epithelial cells). B Expression heatmap and correlation matrix of ENG (fibroblasts) and BMPR2 (epithelial cells). C Kaplan–Meier PFS curve comparing high vs. low expression of the DSC2–DSG2 pair (p = 0.016). D Kaplan–Meier PFS curve comparing high vs. low expression of the ENG–BMPR2 pair (p = 0.017)

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