Background: The infant mortality rate (IMR) is a proxy of the living and health conditions of a given population, which allows us to assess the risk of death for children under one year. Although there is, in general, a reduction in infant mortality in Brazil little is known about this indicator in the regions and cities located on the international borders of the Brazilian territory and the changes that occurred in the face of the migratory impact of the Americas in the period from 1996 to 2020. The objectives of this study are to assess IMR in Brazilian Twin Cities  and its social determinants over time. Methods: This is an ecological study, whose units of analysis were the Brazilian Twin Cities, between 1996 and 2020, based on data on births and deaths in children under one year, available in the public vital information system in Brazil. Data were identified by the city in which the infant death occurred in addition to the mother’s primary city of residence. Correlation measurements were performed to test the associations of the IMR means between the independent variables. Results: The Twin Cities  had higher numbers of infant deaths per place of occurrence than the number of deaths per place of maternal residence. The Northern Twin Cities exhibited the highest IMRs. Cities in the Midwest region showed variability. In the South region, most cities showed low rates. A positive correlation was identified with the Gini index with r = 0.67 and a negative correlation with the Municipal Human Development Index indicator of r= −0.70. Conclusions: The averages of IMRs in the Twin Cities were higher than in their States. In recent years, there has been an upward trend in infant mortality in these cities. The Infant Mortality Rate (IMR) is an important indicator of social well-being, which estimates the risk of death for live births (LB) during their first year of life. It is directly related to the socioeconomic and sanitary conditions of the population. These early deaths could be considered avoidable ,2.The magnitude of the IMR has been associated with the characteristics of the economic development model of a country or region ,4. SinceBrazil has the largest public health system in South America and has an international border with nine countries, namely: French Guiana, Suriname, Venezuela, Colombia, Peru, Bolivia, Paraguay, Argentina and Uruguay , remainiAmong the Brazilian border cities, 33 are considered Twin Cities. They are those that are characterized by urban integration with neighboring countries, with high socioeconomic and cultural flows fundamentally from work, study, consumption and access to public services, especially health and education .The recent wave of migration, seen in different parts of the world, has also occurred in Brazil, with a large influx of refugees already recognized in the country. Of these, 36% are Syrians; 15% Congolese; 9% Angolans; 7% Colombians and 3% Venezuelans ,11. The This is an ecological study with time series analysis (1996–2020), using different vital public information systems in Brazil.The population of this study is formed by all records of LB and infant deaths, of children under one year of age, identified by place of occurrence and residence in the death information record, in the 33 Brazilian Twin Cities.Twin Cities correspond to population densities cut by the borderline . These cities have great economic and cultural potential. There is an intense level of movement of people, goods and capital. ,13. ThesInclusion criteria were all registered infant deaths in children under one year of age, according to the place of occurrence and maternal residence, in the Brazilian Twin Cities, in the period 1996 to 2020. Data on infant deaths over one year of age were excluded of age, of non-border municipalities and of those border cities that are not considered Twin Cities.www.datasus.gov.br) pdf (accessed on 29 April 2021)., using the Mortality Information System (SIM) available on the website  (accessed on 29 April 2021). and the Information System on LB (SINASC) through the address: https://datasus.saude.gov.br/nascidos-vivos-desde-1994 (accessed on 29 April 2021).Data collection was performed through the DATASUS portal  of this system is the Death Certificate (DC) . The SINSocial indicators from the set of Twin Cities were also used, such as demographic data from 2010 and the population estimate for 2019 .The Human Development Index (HDI-M) was extracted from the database of the Brazilian Institute of Geography and Statistics (IBGE)  and was The years 1991, 2000 and 2010 were used from the demographic censuses of the database of the Institute of Applied Economic Research (IPEA) . AdditioThe IMR was calculated from the data available in SIM and SINASC for the place of residence and occurrence using Excel 2010 Software for Windows. Data were tabulated separately by location: residence and occurrence. Data information was descriptively organized in tables and graphs in which the average IMR values of the cities were compared with the averages of the regions. According to the RIPSA  network,Ethical considerations: The study was carried out from a secondary source database, available for public research, whose information is aggregated, without the possibility of individual identification, and is therefore not subject to review by the CEP/CONEP system according to RESOLUTION No. 510, OF 07th APRIL 2016 .During the period 1996 to 2020, in Brazil, 491,761 births were registered by SINASC and 8360 deaths in children under one year of age were reported in SIM, in Twin Cities. Four cities in Rio Grande do Sul  did not present data on the city in which the infant death occurred in addition to the mother’s primary city of residence.While most of the Twin Cities in the North region had the HDI-M classified as medium to low, in the South most cities had HDI-M considered high. In the Center-West region, there was great variability between municipalities, and in the cities of Corumbá (0.700) and Ponta Porã (0.701), the HDI can be considered high. Bela Vista (0.698) and Mundo Novo (0.686) have medium indexes and Porto Murtinho (0.666), Paranhos (0.588) and Coronel Sapucaia (0.589) are low .In general, social inequality is high in all cities observed, with a worsening of the Gini Index over the years, especially in cities located in the North and Central-West regions. However, the Twin Cities in the southern region of Brazil have shown, over the years, an improvement in the Gini index. For example, the Twin City Mundo Novo, located in the Midwest region, which in 1991 had a Gini value of 0.575, in 2010 managed to decrease it to 0.514 .There is an inverse association between FHS coverage and the Gini index. The North region has the largest coverage by the ESF while the South region has the lowest coverage .The North region had the highest IMRs both by occurrence and by residence. For example, we have the rates in the city of Epitaciolândia, which are quite high in the available years. There was an increase in IMR in two cities in the last period, such as Santa Rosa do Purusand Tabatinga. Some IMRs were classified as medium levels . The citIn the Midwest region, there was, in general, a decrease in IMR over the years, with the exception of Paranhos and Mundo Novo. The latter showed an increase in IMR in the last period, from 18.79 to 21.39 deaths per 1000 LB . The SouAmong the cities in the South region that presented available data, most had low IMRs, with the exception of the city of Quaraí, where mortality was higher, with the IMR per residence classified as an average with 21.67 deaths/1000 LB in 2020. Additionally, in the South region, the Twin Cities Foz do Iguaçu-PR, Dionísio Cerqueira and Santo Antônio do Sudoeste showed a stable trend of IMRs due to higher occurrences, being higher than the IMRs per residence .In general, we can observe that there was a decrease in infant mortality. However, in the observed periods of 2014/2015 and 2016/2017, several cities showed an increase in IMRs. In Epitaciolândia—AC, despite an important reduction in the number of infant deaths over the years, the IMRs per residence increased from 2004 to 2005, from 13.38 per 1000 LB to 55.56 per 1000 LB, which meant an increase of 315.24%. In 2010, there was also an increase of 15.35%.A similar pattern occurred in Pacaraima—RR from 2014 to 2015. There, there was an increase from 14.71 per 1000 LB to 16.62 per 1000 LB with a growth of 12.98%. This twin city was decreasing its number of infant deaths in recent years, however, there was a growth of 99.74% in records between the years 2016 and 2017.The same situation can be seen in the city of Bonfim—RR between 2010 and 2011. Their IMR by maternal residence showed an increase of 673.20%. The IMRs due to the occurrence of Twin Cities such as Bela Vista—MS from 2000 to 2001 showed an increase of 29.57%. Between 2016 to 2017, the increase was 31.10%. In Mundo Novo—MS, between 2008 and 2009, it increased by 108.22%, while in Dionísio Cerqueira—SC, between 2010 and 2011, the increase was 29.76% .p-value < 0.001) between IMR and Gini index, with r = 0.67  (The HDI-M showed a negative correlation with the IMR (r= −0.70) and a statistically significant one (< 0.001) . It is nThis is the first study that evaluated IMR in the Twin Cities, after the increase in the recent migratory flow in Brazil. The findings showed that, despite a decline in child mortality in the country, large disparities still exist. The Twin Cities of the North, South and Midwest regions, in 2018–2020, exceeded the infant death rates in their states, with the exception of Porto Murtinho in Mato Grosso do Sul higher than the number of infant deaths by place of maternal residence, indicating an influx of patients to these municipalities, even though these are not regional health care centers. Additionally, an inversion in the profile of the decreasing pattern of mortality was identified, with an increase in IMR in recent years.Some limitations deserve to be pointed out. One of them concerns the use of secondary data, which suffer a delay in their registration with the system and are liable to being under-registered. Furthermore, it is an ecological study, allowing only inferences about populations, which cannot occur at the individual level. The lack of recent census data also impacts the estimates, since it involved the 2010 reference population.Studies by Giovanella and Guimarães, (2007) ; Silva, In Brazil, infant mortality remains a major public health problem, despite decreasing rates over the years. The IMRs of most of the Twin Cities remain higher than those of their states. All Twin Cities had an average IMR higher than the tolerable by the World Health Organization (WHO), which is 10 deaths per thousand LB , in the However, the rates show inequalities even in those cities with lower IMRs in the South region, followed by the Midwest. This finding is similar to those reported by Peiter (2005)  and Souz33, which resulted in an improvement in the Gini index. The Twin Cities belonging to the North region, on the other hand, have more precarious living conditions and worse access to health services [Another important aspect may be the cross-border dynamic, due to the economic integration project promoted by Mercosurservices ,30,31,32The main causes of death in children under one year of age in the municipalities belonging to the border zones were neonatal/perinatal causes resulting from poor care for pregnant women (22.8%) and preventable deaths due to lack of care for the newborn. (17.3%) . DiffereAn increased risk of death from preventable causes in children living in the Twin Cities had already been identified by Souza and Silva (2018) . HoweverThe worsening of living conditions and the lack of access to medical care, which may be caused by cuts in social investments and in the SUS  in recent years, were associated with an increase in mortality in specific categories ,36. AddiSome border municipalities had IMRs per place of occurrence greater than the number of deaths per place of maternal residence, indicating an influx of patients to these cities. Most of the Twin Cities are small and do not have much support in terms of health care.The recent wave of migration observed in different parts of the world has also occurred in Brazil, with a large increase in refugees. In 2020, 82,520 permanent visas were requested, 65.1% of which were Venezuelans, followed by 20.1% Haitians and 4.8% Cubans . The TwiThe study by Lima et al. (2020)  found thDemographic, economic and cultural characteristics in border municipalities can directly lead to the probability that a child reaching his first year of life is greater than that of the group of non-border municipalities  Infant mThe pattern of increased mortality in recent years in the Twin Cities, as well as the increase identified in some specific categories, is also associated with worse living conditions in these cities. The HDI-M, which measures the well-being of a population, especially children, had lower averages in the Twin Cities (0.674) than the Brazilian average (0.727), with the exception of Foz do Iguaçu (0.751) .These cities have deficiencies in education, health deficiencies, and worse employment and income, in general, when compared with other non-border Brazilian municipalities. Additionally, the border strip is marked by the contradiction between what is legal and what is illegal, attracting groups that use illegality to entice young people and women as mules of trafficking; men for the crossing of drugs, weapons, goods, etc. ,42.Social inequality, measured by the Gini index, is also an important factor for infant mortality, showing a direct correlation with this in the Twin Cities. The regions with the highest mortality are also those with the highest indicators. Municipalities in the South region, which in general have lower mortality rates, also have lower Gini indexes . The TwiAccording to Guimarães et al. (2016) , the SouThe care model focused on primary care and family health has been associated with the improvement of health indicators such as, for example, low birth weight and lower infant mortality) ,44. One The Twin Cities in the North region have the largest coverage of the FHS. Poorer regions such as the North and Northeast, which have the highest levels of poverty in the country, also have the greatest coverage . The intWe show that most of the Twin Cities have high infant mortality rates, not only higher than recommended by the World Health Organization (WHO) but higher than the states where they are located. In addition, we identified that there is a change in the pattern of IMRs in these cities, which show an increasing trend of infant mortality in recent years in cities belonging to the North region, followed by the South region. Recent changes in the mortality pattern with mortality rates per occurrence greater than those of residents in border towns without a specialized assistance network may suggest a search for these towns by migrants. The hypothesis of an association between recent migratory flows and infant mortality needs to be explored through other study designs and deserves the attention of managers and researchers.There is an important gap in information about infant mortality in the Twin Cities and the migrant population, as well as the impact on the health systems of these cities. This information gap also occurs in other Latin American countries. These cities have different characteristics that demand greater attention in terms of surveillance and health care, caused by the rapid and, at times, intense flow of people, and that need to be taken into account by health managers.


Background: Melanoma genetic testing reportedly increases preventative behaviour without causing psychological harm. Genetic testing for familial melanoma risk is now available, yet little is known about dermatologists’ perceptions regarding the utility of testing and genetic testing ordering behaviours.Objectives: To survey Australasian Dermatologists on the perceived utility of genetic testing, current use in practice, as well as their confidence and preferences for the delivery of genomics education.Methods: A 37-item survey, based on previously validated instruments, was sent to accredited members of the Australasian College of Dermatologists in March 2021. Quantitative items were analysed statistically, with one open-ended question analysed qualitatively. Results: The response rate was 56% (256/461), with 60% (153/253) of respondents between 11 and 30 years post-graduation. While 44% (112/252) of respondents agreed, or strongly agreed, that genetic testing was relevant to their practice today, relevance to future practice was reported significantly higher at 84% (212/251) . Ninety three percent (235/254) of respondents reported rarely or never ordering genetic testing. Dermatologists who viewed genetic testing as relevant to current practice were more likely to have discussed (p < 0.001) and/or offered testing (p < 0.001). Respondents indicated high confidence in discussing family history of melanoma, but lower confidence in ordering genetic tests and interpreting results. Eighty four percent (207/247) believed that genetic testing could negatively impact life insurance, while only 26% (63/244) were aware of the moratorium on using genetic test results in underwriting in Australia. A minority  reported prior continuing education in genetics. Face-to-face courses were the preferred learning modality for upskilling.Conclusion: Australian Dermatologists widely recognise the relevance of genetic testing to future practice, yet few currently order genetic tests. Future educational interventions could focus on how to order appropriate genetic tests and interpret results, as well as potential implications on insurance. Availability of genetic testing for hereditary cancers has increased in the last 2 decades . HoweverCDKN2A, accounts for 20%–40% of hereditary melanoma cases and 90% of positive results  practitioners , obstetrA cross-sectional survey was posted to all Australian Dermatologists to capture current confidence and attitudes regarding genetic testing, and guide future education interventions. Specifically, we were interested in predictors of genetic confidence, and the relationship between attitudes towards genetic testing and current practice.The capability, opportunity, and motivation model of behaviour (COM-B) was the guiding framework in the survey design. The COM-B model considers three interacting constructs relating to the uptake of a new practice including; 1. Capability (knowledge and competence), 2. Opportunity (resources), and 3. Motivation (perceived benefit of genetic testing) . This stBRCA1 or BRCA2 testing, and selecting questions relevant to the potential benefits and limitations of genetic testing for familial melanoma (Agree/Disagree); a 5-item scale  and ratified by the University of Technology Sydney HREC (ref: ETH20-5671).In March 2021 a survey was posted to all Australian members of the Australasian College of Dermatologists (ACD), excluding trainees as no site of employment was available for them. Contact details were accessible for only one member in New Zealand, therefore only Australian members were included. A paper-based survey was administered, to allow the inclusion of an AUS$10 cash incentive, as incentives have previously been shown to increase physician response rates . The ACDp-values of <0.05 were considered significant and reported with 95% confidence intervals (CI). Qualitative responses to the open-ended question were evaluated using thematic content analysis methods  reported having previously discussed it; 91% had offered it, 89% had referred patients for it, and 60% had previously ordered tests. When asked how frequently genetic testing was ordered, 29% reported never ordering, 63% ordered rarely (≤5 times a year), 6% often (≥ once a month), and only 1% (n = 3) routinely (≥ once a week). Only 19% indicated they had the “necessary services and staff” to offer genetic testing. Respondents who indicated they “never” or “rarely” ordered genetic testing, were subsequently asked, “why not?”, and provided with a list of common reasons to select from, including an open field for “other”. Of note, 22% of respondents reported that their lack of confidence contributed to their infrequent ordering. Results are displayed in t = -9.82, p < 0.001).While 44% of respondents agreed/strongly agreed that genetic testing was relevant to current practice, 85% agreed/strongly agreed that genetic testing will be relevant to future practice  or offered it to patients, X2  = 17.5, p < 0.001, ordered testing, X2  = 6.8, p = 0.009, and referred patients to genetic services, X2  = 9.3, p = 0.002.Participants who viewed genetic testing as relevant to their current practice were significantly more likely to have discussed , and identifying patients at risk of a hereditary condition . Moderate confidence was reported for identifying patients who could benefit from genetic testing , educating patients on genetic aetiology of disease , obtaining informed consent , discussing possible outcomes of genetic testing , and using genetic information in management decisions . Lower confidence was reported for identifying appropriate tests , test ordering , and interpreting results . A cumulative average was calculated to provide an overall genetic skills confidence score . Confidence results are displayed in Prior experience with continuing education in genetics, and its relative usefulness are displayed in Respondents were asked to rank their preference from 1 to 5 (with one being the most preferred option) for the delivery of future education initiatives in genomic medicine. For the 141 who completed this question correctly, the most preferred learning modality was face-to-face courses (42/141), followed by a hotline for genetics advice (38/141), online courses (34/141), experiential (17/141), and lastly, printed material (10/141). When averaging the ranked score for each modality the order of preference changed only for online courses (third to second preference) and hotline for advice (second to third preference) .Linear regression was used to identify independent variables associated with dermatologists’ overall confidence in their genetic skills .p = 0.007). Perceived relevance of genetic testing to current practice significantly predicted confidence , however perceived relevance to future practice was not significant. Genetic confidence was significantly predicted by having previously discussed , offered , ordered , or referred patients for genetic testing . When continued education variables were assessed, an “award unit of study on genomics’ was not associated with overall confidence, but completing a short course  or receiving training on ethical considerations  significantly predicted genetic confidence.No significant relationship was found between years since graduating from medical school, however having a sub-speciality significantly predicted genetic confidence  identified the need for further training, education, and/or resources for providing genetic testing in clinical practice. Specific suggestions included requests for the College (ACD) to provide/advertise a short course on genetics, presentation updates at the Annual Scientific Meeting, and online Continuing Professional Development modules. There were several recommendations for a “cheat sheet”, or “flip book” providing clinicians with core information on tests, testing criteria, genetic counselling discussion points, test ordering and result interpretation. A few respondents expressed a need for information on relevant costs and Medicare rebates for genetic tests. Twelve individuals indicated that they are already referring patients for genetic testing, with three respondents expressing a preference to continue to refer patients rather than provide genetic testing directly. Ten comments stated the need for greater access to genetic services. Just five individuals commented that genetic testing was not useful or needed in dermatology. Five further respondents expressed concerns about costs, genetic discrimination regarding insurance, and concerns of patient anxiety.Almost a third of participants (n = 81) provided a response to open-ended question To our knowledge, this is the first cross-sectional survey to comprehensively examine how dermatologists are currently using genetic testing in practice, including confidence in ordering genetic tests, attitudes towards the utility of testing, and preferences for education and training. Respondents were more likely to believe genetic testing would be relevant to future practice than it was currently. This finding is consistent with a recent study of Australian non-genetic clinicians which reported that genetic testing would become increasingly important in the future . FurtherQuestions exploring perceptions regarding genetic testing identified a knowledge gap regarding the potential impact on insurance in Australia. Less than a third of respondents were aware of the 2019 Australian moratorium on the use of genetic testing results in insurance underwriting . FurtherThe first construct in the COM-B model relates to capability, that is, possessing the necessary knowledge and skills to perform the task . This suAny training initiatives for dermatologists to provide genetic testing would need to cover the challenges of informed consent and interpreting and explaining test results. If a dermatologist has a patient test positive for a genetic test, they are advised to report this result to the patient and then refer them to clinical genetic services for further discussion regarding appropriate screening for non-cutaneous cancers and to arrange testing for at-risk family members. Any individuals found to carry mutations through this testing process would then be referred back to the dermatologist for appropriate screening.An important objective of this survey was to understand the impact of past genomic education/training to guide future interventions. It was encouraging to find that completing a short course in genomics or receiving training on ethical considerations of genomic research positively predicted overall confidence in performing genetic tasks. While completing a unit/subject as part of an award course was not associated with confidence, this is not unexpected, as such genomic units are usually undertaken during early undergraduate degrees, and therefore were more than 2 decades ago for most respondents. Preferred learning modalities for future genetic education included face-to-face training, a hotline to a genetic professional, and an online course. Face-to-face courses provide opportunity for group training in an interactive setting and have previously demonstrated positive impacts on attitudes and behaviours . A genetCapability, Opportunity, and Motivation) are imperative to the successful implementation of new interventions. The survey results have clearly demonstrated motivation for upskilling in this area by dermatologists. Furthermore, results on confidence in core genetic testing tasks are useful in guiding emphasis in training materials to boost clinician capability. However, training initiatives are only useful if there is a similar focus on opportunity—this may be difficult given the majority of respondents report having insufficient resources to offer patients genetic testing. Future education and training interventions for mainstreaming genetic testing into dermatological practice would benefit from comprehensive consideration of factors affecting opportunity . This is comparably higher than some recent genomic medicine surveys to non-genetic health professionals (11%–26%) . HoweverAustralian dermatologists perceive genetic testing as increasingly relevant to practice. However, they currently report low confidence in ordering tests and interpreting results. Having received continuing education on genetics/genomics was found to positively predict genetic confidence. We also found a strong demand for additional resources to enable the upskilling of dermatologists in how to provide genetic testing. This study has generated new evidence to help inform the future implementation of genetic testing into routine dermatological practice in Australia.


This article defines "cultural experience" and places it in a holistic conceptual model; “the cultural city” where it plays a relevant role in improving the performing of cities. The conceptual model combines the basic elements of the heritage city, the smart city and the creative city. The city is interpreted from a threefold perspective; as a repository of resources, as a connective interface, and as the setting for citizens' life and social and professional experiences. In this context, each of these perspectives incorporates culture in a different way, enabling different models of value creation and different processes of production and reproduction of this value. In each of the urban models described above, production processes that combine symbolic, physical, financial, social, human and cultural capital in different ways and urban strategies are implemented to provide cultural experiences that ignite transformative effects through several spillovers. That means that culture, in its different dimensions, regains the role of a raw material and becomes the point of origin to activate development processes and improve urban performance. The integration of the dimensions of the heritage city, the creative city and the smart city through an enabling context is the core proposal of the “cultural city”. In alignment with the New European Agenda for Culture, we deepen the analysis in the specific spillovers on wellbeing and quality of life, citizen engagement and urban renewal as the backbone of a set of external effects of cultural experiences. In the final part of this article, we test the plausibility of this speculative proposal through some empirical evidence. We develop an OLS model with proxy indicators, that could be considered transitional indicators, for the three different potential strategies . The findings support the assertion that it is conceivable that the supply of cultural experiences through a variety of tactics  can account in part for the growth of European cities in the years after the 2008 financial crisis. These strategies have contributed to the good performance of the urban device in a way that is positive, not negligible (accounting for around 50% of the variance in productivity) and statistically significant. The provision of a context that increases the cultural experiences for citizens has clearly improved the performance of European cities, and we develop some conceptual and empirical mechanisms to explain and measure the socioeconomic impacts of these processes. There is no doubt that the city, as a device for human interaction and a mechanism for generating wealth, has been remarkably successful. The key to the city's success and persistence lies in the fact that it satisfies human needs with high efficiency levels, and when it does not, mechanisms appear to generate the necessary changes to transform itself. As Jane Jacobs stated more than 50 years ago, "Cities contain the seeds of their own regeneration” Jacobs . The ver(Manifesto of the Communist Party), and it was also in the industrial cities that the dream of a new social order was forged and reinforced. The city has always been the neural centre of freedom, culture, and political and institutional innovation in its broadest sense. The exchange of ideas and experiences, the cultural "mix" that is consubstantial to cities, has meant an enormous positive externality for society as a whole, to the point of Jane Jacobs' affirmation "The city, the wealth of nations", which perfectly summarises this powerful idea.As the Shanghai Manual , material  and technological contexts, could become a very powerful tool for social transformation, influencing the general model of urban performance, including its economic framework.However, if we are interested in delving into the impacts of culture beyond its economic classification, we will have to look at the impact generation process, considering the concept in all its complexity. What we seek with this approach is an operational definition of the basic process that activates and generates these processes of transformation and change in order to try to articulate plausible sequences of causality of these impacts. In his early work, Matarasso  spoke ofresonance on our cognitive, emotional or aesthetic dimension or our perception of our location in a social body. A cultural experience is a concrete act of cognitive, sensory and emotional appropriation of the world around us, the intensity and quality of which depends on material, psychological and social issues, as well as on our own cognitive and cultural capital.A “cultural experience” can be defined as the generation, emission or reception of information flows with symbolic content, usually expressed through artistic grammars, that have the explicit and more or less deliberate intention of having some kind of resonance from the German philosopher and sociologist H. Rosa, who states  theThese dimensions of the city are intertwined and articulate its social, political, symbolic, and economic fabric, as shown in the conceptual map below. According to our hypothesis, the activation of cultural experiences through any of these mechanisms will generate social, cultural and economic value, consequently improving the efficiency of the city as a "social artefact" to a greater or lesser extent. In the final part of this article, we make an instrumental simplification and assume that the greater efficiency of the "urban engine" can be approximated through an indicator such as the variation in productivity. The view we defend is that since the 2008 crisis, European cities have used some of these strategies, with varying degrees of instrumental rationality and intuition, to improve urban efficiency Fig. .Fig. 1CoThe "cultural city" as a space and support for the cultural experiences of individuals becomes a relevant variable to explain the success of cities. There is extensive literature from different disciplinary fields on the location of culture and creativity in urban complexes. From the late 1980s until the onset of the crisis, different theories successfully pointed, specifically and in a renewed way, to the cultural dimension of cities Zukin  as an opCulture—understood as the set of cultural experiences that are activated in a given territory over a period of time—is interlinked and generates value in different ways, which are described in the following paragraphs.The first perspective from which the concept of a city can be approached is as a geographical space where a large number of resources are concentrated Scott . This laIt should be noted that urban assets, although they refer especially to the material dimension of the city, are not limited to the physical artefacts that make up cities, such as the grid of streets, buildings, gardens, monuments or public and private facilities. These should be added to the value of the iconic elements and the stories or meanings associated with the material elements. In this sense, the city can be seen as a container for the meanings attached to its material contents where the capacity to generate value is often much more related to the discourses than to the physical elements. In post-industrial cities Scott , value isense of place. Moreover, such discourses are a constituent part of the cultural and cognitive capital of the people who inhabit, use or visit the sites and consequently condition their behaviours and ways of relating to each other and to the space  , where competing interests and alternative use of resources and patterns of appropriation of public and private spaces are settled, and spaces of communication (collaboration). Density is both an agitator of conflict and a fertiliser of communication. “One of the key elements in this context is that the city enables the concentration of human capital, which as we know from the Romer-Lucas models Romer  is the cAgglomeration economies are the result of both economies of scale and the network economies that develop when firms and people are located close to each other. They are therefore related to spatial proximity and, Glaeser,  states, The third dimension to which we wish to refer is the concept of the city as the setting for the vital, personal, professional and social trajectories of the people who inhabit it. With urbanisation levels expected to reach 70% by 2050  and the lived city (la cité) , whose final objective is the maximization of residents’ well-being Perucca .The interplay between urban and territorial renovation, culture and cultural initiatives and urban governance modes  and risk-proneness (proxy for innovation).However, the purpose is not to elaborate a complete econometric model capable of fully explaining the economic growth of cities, but to determine whether these elements have a significant impact and, if so, whether it is a positive one. This is a first empirical approach to confirm or disprove our hypothesis. We are aware that there are already numerous studies in the academic literature on productivity growth that are extremely accurate and that incorporate variables such as the capital stock, the rate of capital depreciation, the rate of growth of technology, etc. These variables, however, are hardly obtainable in a reliable way at the local level. While all these additional variables, among others, would be necessary for a rigorous sophisticated model that attempts to explain productivity growth accurately and robustly, this is not the purpose of this article. We would like to stress that the added value of this work does not lie in the robustness of its empirical evidence but in the consistency and plausibility of its theoretical proposal.i, the estimated increase in productivity depends on the sum of its indicators in the areas of heritage, creativity and smartness as explanatory variables, multiplied by their coefficients  contribute to their economic growth and development and, if so, to what extent. As a proxy for the concept of economic growth, we will use the cumulative change in productivity between 2008 and 2018. An ordinary least squares regression (OLS) is applied, with the following equation:The indicators defining the heritage, creative, and smart components are explained in the following section.Obtaining indicators and comparable data at the local level is always a major challenge, as they are not always accessible and sometimes, they do not even exist. To address this problem, the database has been built using a combination of different sources. However, a limitation of this method is that the city coverage of the different indicator panels does not always coincide. The Cultural and Creative Cities Monitor . Based on these data, the indicator used in the model corresponds to the cumulative change, in percentage points, between 2008 and 2018.Heritage, Creative and Smart.Heritage is composed of a single indicator: museum visits per 1,000 inhabitants, which is obtained from the Cultural and Creative Cities Monitor. The original source is Eurostat (Urban Audit) and the data refers to the period 2011–2017.Creative is composed of two indicators:New jobs in creative sectors per 100,000 inhabitants. Derived from the Cultural and Creative Cities Monitor, and originally collected from Eurostat . It includes three sub-indicators that are weighted equally: new jobs in arts, culture and entertainment enterprises; in media & communication; and in other creative sectors. The data corresponds to the period 2010–2016 and to the NUTS 3 regional level in which each city is inserted.Willingness to take on risk, defined as the percentage of people who disagreed with the statement “One should not start a business if there is a risk it might fail". It is taken from the European Digital City Index, which in turn takes this indicator from the 2013 Eurobarometer. The data correspond to the NUTS 2 regional level in which each city is inserted.Smart is also composed of two other indicators:Commute. This variable is also derived from the European Digital City Index. It is a score that is calculated from Numbeo data and considers the average distance and travel time from home to work. Higher values represent better scores, i.e., shorter time and shorter distance, showing a better performance of the city as an interface device.Annual graduates in ICT per 100,000 inhabitants. Derived from the Cultural and Creative Cities Monitor, and originally collected from the ETER project. Data corresponds to the period 2013–2015 for tertiary education.There are three explanatory variables, which we have called Given the difficulties of obtaining standardised data at the local level, these indicators have been chosen because they reasonably capture some of the defining features of the conceptualised city typologies. We select visits to museums not only because museums are the most representative repositories of the heritage stock of cities in its many different forms, but also because the number of visitors is a good indicator of the enhancement of this heritage and the involvement of citizens in it. Moreover, both the emergence of new creative jobs  and the willingness and open-mindedness of the population to take risks and undertake uncertain projects, ideas and initiatives (as a prerequisite for developing creativity and innovation) are variables that allow us to quantify the capacity of cities to activate creative processes. Finally, a highly digitalised environment with widespread access to and use of technological tools  and efficient transport infrastructures that minimise the time and distance between places and allow cities to become accessible spaces of interpersonal connection are two defining features of Smart Cities, as explained above.All the raw indicators used to construct the explanatory variables, both those from the Cultural and Creative Cities Monitor and from the European Digital City Index, are first standardised according to population. Subsequently, they have been subjected to a winsorization process in case they contained outliers. That is, if the distribution of a variable has a kurtosis greater than 3.5 and an absolute skewness greater than 2, upper-end outliers are substituted with the next highest value and lower-end outliers with the next lowest value. This process is repeated iteratively until a distribution that meets the kurtosis and skewness requirements is obtained.i, i, and min(x) and max(x) are the minimum and the maximum value in the sample for variable x.This process of winsorization is followed by a min–max normalisation process, so that all indicators fall within an interval of 0 to 1. This, in addition to allowing a direct comparison of the coefficients of the three components considered in the regression , is necessary to aggregate variables with different magnitudes within the same score. It also applies in the case of the new creative jobs variable. This, as mentioned above, in turn considers three different indicators: new jobs in arts, culture and entertainment enterprises; in media & communication; and in other creative sectors. In order to weight these three areas equally, so that the different dimensions do not introduce biases, the min–max score is obtained first, and then averaged. The scores from the Cultural and Creative Cities Monitor and the European Digital City Index have not been used directly but have been recalculated for the sample of cities used in our analysis. The equation used for the min–max normalisation is as follows.Table The results of the OLS regression applied following Eq.  are show2 of 0.5034. This means that the model only explains about half of the variability in productivity growth of cities. The limited explanatory power of the model must therefore be borne in mind. However, this should not come as a surprise. It is worth remembering that the aim is not to determine all the factors that contribute to productivity growth in cities from a holistic perspective, but only to test the effect of some of them, i.e. those conceptualised in this article. Naturally, the model leaves out a multitude of explanatory factors, ranging from the national and regional economic context to the productive structure, the embedded capital or the provision of key infrastructures.The OLS model has an adjusted RNevertheless, given the enormous complexity of the phenomenon under study and the multitude of factors that are not considered, the model is still a reasonable fit. Figure heritage = 1.027, creative = 1.059, smart = 1.069), so the presence of multicollinearity is discarded.The validity of the model is tested by applying a Shapiro–Wilk normality test  and have also found an indicative way to measure their effects and test their plausibility. The conceptualisation of the “Cultural City", which integrates all three approaches, opens up new avenues for research and comparison in other geographical spaces, other scales and other periods.We have sufficient evidence that culture and creativity have played a relevant role in the recovery of European cities in the aftermath of the 2008 crisis. This effect has been articulated in different ways. We have been able to define a conceptual structure that includes the three main strategies From the point of view of policy recommendations, increasing the provision of cultural experiences is a strategy that improves the performance of cities. The second recommendation is that the social values generated through the Heritage City can be enhanced and formulas beyond tourism should be sought, and the third is that digitalisation and the improvement of urban switching speed, both of which are quite dependent on local authorities, have a considerable impact that is likely to become even greater as a result of the Covid-19 pandemic.It seems that we can accept that the analytical approach we set out in the first part of this article is plausible. It is plausible that part of the growth of European cities in the post-2008 crisis period can be explained by the provision of cultural experiences through different strategies . These strategies have statistically and positively contributed in a significant way to the good performance of the urban device, accounting for around 50% of the variance in productivity. The interpretative framework that we have called "the Cultural City" represents a more or less balanced combination of the Heritage City, the Smart City and the Creative City. We are of course aware that we are not dealing with a complete and definitive test that validates this new framework. Rather, we are making an approximation to the plausibility of the proposal through partial and circumstantial evidence that so far fits.In a way, we are identifying some transitional indicators that make it possible to connect cities’ cultural experiences and performance improvement processes. It must be understood that although our dependent variable is the variation in the productivity of the labour factor in the cities, this variable approximates the good performance of the cities and includes the impacts on different dimensions that beyond the strictly economic . Thus, variables such as risk propensity, the number of visitors to museums or the number of ICT graduates anticipate the fluidity of the transmission processes between cultural experiences and the impacts on good urban performance. These transitional indicators are not limited to those that fit statistically into the model (or are available to us) but point to a wider family of variables that enable the transformation processes that take place through cultural experiences. These transitional indicators need to be investigated more intensively, as they define the transmission mechanisms between the policies and projects that produce cultural experiences and their final impacts on the economy, culture or society.Although the ways of generating value are very diverse, the main ways in which we should focus in future research are those that materialise through the improvement of citizens' health and wellbeing, those that are generated by a greater commitment to the community , and those that materialise in the processes of urban regeneration with social impacts through “placemaking” processes and economic value generation, placing high added value activities in new refurbished urban spaces or through real state or tourism impacts. Another possible area of research improvement would be to connect individual preferences with cultural experiences by testing their effects on socio-economic impacts. This research will be further developed in the future with the AU Culture application that tries to measure individual impacts of cultural participation (see the Resources of the MESOC project).If we look at levels, it seems that the Smart dimension is the one that has contributed the most to growth, with an impact that is twice that of the Creative dimension and almost three times that of the Heritage dimension. These differences probably have to do with the times and circumstances we are living in. Since the 2000s, cities have invested in technology to enhance their competitiveness. One of the thematic objectives of EU Cohesion Policy during the 2014–2020 period was to enhance access to, and the use and quality of information and communications technology, including developing products and services and strengthening applications. The EU eGovernment Action Plan (2016–2020) currently sets out concrete actions to accelerate the implementation of existing legislation and the related uptake of online services. The digital transition is reshaping public services, and it is clear that its impact is very significant. Nine out of ten cities report that their services have improved as a result of digitalisation ESPON . ChangesWithin the scope of the Creative Cities strategy, we can identify large European capitals that are also major centres of creativity and culture such as Paris, London, Madrid, Amsterdam or Copenhagen. Finally, the Heritage strategy is more prevalent in cities with significant historical and artistic heritage such as Rome, Lisbon or Ljubljana. The reason why the Heritage City strategy has a lower impact on the model is probably because the way to capitalise the impacts is either through tourism, an activity with low average productivity, or through the increase of real estate value, an activity that has gone through a crisis during the period considered.In conclusion, the provision of contexts that increase citizens’ cultural experiences has clearly improved the performance of European cities and this study suggests a series of conceptual and empirical mechanisms that can help explain and measure the socioeconomic impacts of these processes.


Recent haplotype sharing analyses in specific European populations have revealed fine-scale genetic differentiation that echoes history. An equivalent understanding across the whole European continent would place these insights into a wider context and extend understanding to underdescribed regions. Here, we leverage haplotype data from 5,500 European-ancestry individuals from the UK Biobank (UKBB) in a methodological approach to update and expand the European genetic landscape, extending coverage to regions in the southeast of Europe, identifying communities of high haplotype sharing that may be of interest to genetic mapping, such as Malta. Together, our results highlight European communities with diverse ancestries sampled within the UKBB and demonstrate the potential for insights to be made with other non-European ancestry communities using this dataset. Haplotype-based analyses have recently been leveraged to interrogate the fine-scale structure in specific geographic regions, notably in Europe, although an equivalent haplotype-based understanding across the whole of Europe with these tools is lacking. Furthermore, study of identity-by-descent (IBD) sharing in a large sample of haplotypes across Europe would allow a direct comparison between different demographic histories of different regions. The UK Biobank (UKBB) is a population-scale dataset of genotype and phenotype data collected from the United Kingdom, with established sampling of worldwide ancestries. The exact content of these non-UK ancestries is largely uncharacterized, where study could highlight valuable intracontinental ancestry references with deep phenotyping within the UKBB. In this context, we sought to investigate the sample of European ancestry captured in the UKBB. We studied the haplotypes of 5,500 UKBB individuals with a European birthplace; investigated the population structure and demographic history in Europe, showing in parallel the variety of footprints of demographic history in different genetic regions around Europe; and expand knowledge of the genetic landscape of the east and southeast of Europe. Providing an updated map of European genetics, we leverage IBD-segment sharing to explore the extent of population isolation and size across the continent. In addition to building and expanding upon previous knowledge in Europe, our results show the UKBB as a source of diverse ancestries beyond Britain. These worldwide ancestries sampled in the UKBB may complement and inform researchers interested in specific communities or regions not limited to Britain. Recent studies utilizing haplotype- (or “chunk”-) based approaches  have dem2At the same time as improvements into the study of genetic history with haplotype-based coancestry information , 10, 11,171)What sample of European genetics is found in the UKBB dataset?2)Utilizing chunk-based haplotype methods, how does the genetic structure found within this large European sample match and inform on previous European population genetics research?3)What is the diversity of demographic histories captured in this sample of European haplotypes?Motivated by recent haplotype-based analyses of specific European populations –4, 7, 23Investigating the genetic landscape of Europe, we subsetted individuals form the UKBB  who sampSI Appendix, Supplementary Data 2), with samples from most regions forming a primary cluster reflective of their common ancestry. Some regions appear more heterogenous such as Germany or Malta , suggestive that a substantial fraction of individuals in the dataset have a recent genetic ancestry that does not match most individuals with the same place of birth label, possibly due to modern economic or other recent migration within the continent. Due to this heterogeneity in birth-location label versus principal component (PC) coordinates, using a separate PCA, we projected our European sample to western Eurasian references from the Human Origins dataset . This suggests a community of individuals with shared common ancestry that is not private to one European country but is more common in samples from Eastern Europe. To further explore the possibility of genetic communities not well captured by country-of-birth-based labels, we sought to cluster individuals based on degrees of haplotype sharing.Interestingly, there appears to be a common cluster of individuals projected onto the same PC space who collectively do not match the ancestry of individuals with their reported place of birth. A substantial number of individuals of Hungarian, Czech, or Russian birthplaces appear to be members of this cluster. Indeed, these individuals separate together on PC six , visualized  the structure captured in the pbwt coancestry matrix by performing PCA , and compared clusters to Human Origins references . Adapting an approach using the “non-negative-least-squares” function . Finally, we also assessed intercluster distance using total-variance-distance estimates (Dataset S2) and FST (Dataset S3) using “ADMIXTOOLS2”  value of 0.0006 (Dataset S3) and more limited dispersal in PC space , France, Switzerland, the British Isles, and Ireland. Differentiation is low within this branch, as evidenced from an average within-branch fixation index  from the others. All 17 Icelandic individuals sampled are branched with Norwegians, which we attribute to the small sample of Icelandic individuals. In the nnls analysis we observe that the Netherlands and Denmark clusters show evidence of haplotype sharing consistent with their geographic proximity.ST and total variance distance (TVD) values (Datasets S2 and S3). The majority of Estonian individuals are placed in the “Mixed Scand.” cluster that also groups Swedish, Norwegian, and Finnish individuals who project between Finland and the rest of Scandinavia.The CE Europe branch contains the following three subbranches: NE Europe , CE Europe , and Finland as an outgroup. Consistent with previous observations , 35 FinlIndividuals from the Baltic countries Latvia and Lithuania are clustered together. Estonian individuals project intermediately between Finnish and Baltic–Russian individuals in PC space , suggestThe CE Europe branch bridges northeastern and western Europe in PC space , containSI Appendix, Fig. 4.3).The two clusters of northern Balkans membership include 246 of the 448 individuals from the Balkans/SE Europe region . Cluster membership is predominantly from countries north of Albania/Greece/North Macedonia—suggesting a north/south divide on the peninsula that is echoed in PCA . The “NWLeiden clustering yields three broad groups of clusters with southern European membership, as follows: one grouping individuals born around the eastern Mediterranean , the Iberia Peninsula , and finally two outgroup clusters .SI Appendix, Fig. 4.3).Most sampled Greek individuals form a single cluster that in the nnls analysis is modeled as a mixture of haplotypes from neighboring southern clusters such as “Italy” and “Turkey,” but as well haplotypes from the north from NE Balkans. We observe a smaller cluster grouped with the “Greece” cluster on the dendrogram  that conn = 200). In PCA, these are distributed along a gradient along PC seven, forming three clusters. The first contains Maltese individuals who projected separately from the rest with British samples. The other two project away from these Maltese samples, with one equidistant between the British–Maltese individuals and another of sole Maltese membership in PC space . Our Leiden clustering groups these latter 2 groups of samples into 1 cluster of 79 individuals and the former into clusters of predominant British membership. These results suggest recent western European gene flow into Malta, with the intermediate PCA cluster of Maltese individuals consistent with first-generation offspring of ancestral Maltese and western European parents. Nevertheless, we identify a core cluster of Maltese membership who project onto the same PC space as Human Origin references and with who we investigate the Maltese demographic history below.We report to our knowledge the largest sample of dense genome-wide genotypes from the small Mediterranean archipelago of Malta  and found strong evidence that this cluster indeed reflects a community with Ashkenazi Jewish ancestry.Finally, we observe that Leiden clustering identifies a cluster whose membership includes the individuals who separate out along PC six of the PLINK-based PCA. As describe above, these individuals have a mixture of European birthplaces, with a slight bias toward central or eastern Europe. Previous work has shown evidence that the British Jews have been sampled by the UKBB . To test sharing  . Complementing these IBD-based approaches, we estimate inbreeding coefficients (ROHF and SNPF) to explore the evidence of small-effective population size versus endogamy  and focusing on subbranches of related clusters , and the inbreeding coefficient that is measured by the observed versus the number of expected homozyotes (SNPF) for the Finnish clusters  from those characterized by large numbers of short IBD-segments . For individuals within the same cluster, we plotted the per-individual mean total length of IBD versus the mean number of IBD segments shared , showingclusters B. Generaclusters , 39, as analysis  and averclusters .e) and low levels of haplotype sharing , there are notable exceptions. Maltese genetic differentiation is expanded upon in IBD-segment analysis , we conclude that these represent a distinct community or population clustered with other Spanish individuals who nevertheless exhibit elevated haplotype sharing consistent with isolation. These individuals project from other Spanish individuals in PC six  and when projected on top of Human Origin references project toward “Spanish North” or “Basque” references .Within SE Europe, both Turkey and Cyrpus exhibit elevated haplotype sharing , as welle  and evide . The IBDe . Albaniain Spain . In a foe 10 generations ago  or number of segments .Continuing previous observations of elevated haplotype sharing in island populations, we confirm previous signatures of isolation in island communities in northern Britain and expand with more results. We observe increased haplotype sharing of the British archipelago communities of Orkney, the Channel Islands, and the Isle of Man  that is ions ago , as wellions ago , there iions ago . We obsee 30 generations ago that has expanded within the past 10 generations . Worldwide ancestries have been shown to be captured by the UKBB , and thiMalta cluster, and another projecting in between the former two. We therefore already identify recent migrants to the island who do not represent the historical ancestry there. The collective genetic profile of the Malta cluster is also consistent with previous literature, where Maltese uniparental data . Our results, in the context of disease mapping work highlights the potential of this isolate community in genetic mapping efforts. As similarly argued recently by Borg et al.  consists of Albanian, Kosovan, and some North Macedonians. Whether this is a specific immigrant community sampled by happenstance in the UKBB or a feature of the region is unclear, and expanded sampling within this region may further elucidate how varied the haplotype landscape of this region is.We improve sample coverage in SE Europe where sampling has been lower than other European regions in previous analyses –54. Our SI Appendix, Supplementary Data S7, to summarize, we show using f-statistics . The demographic profile from haplotype sharing and estimated historical Ne of individuals within this cluster matches well with a proposed demographic model from a previous study of European Ashkenazi Jews .Elsewhere in S European genetics, we have studied a genetic community of individuals that we denote as Mixed European. Although we discuss in more detail in atistics  that theazi Jews  with a bazi Jews , which mOverall, our work has demonstrated a utility of large cosmopolitan biobank studies in providing an informative continental sample of European genotypes. We have leveraged this to expand the map of the European genetic landscape and show genetic signatures of interest to geneticists. The use of joint analysis of both chunk- and IBD-based haplotype analysis shows that they complement each other to interrogate the population structure and demographic profile of different communities on the continent. This work raises the possibility of similar analyses of non-European communities within the UKBB, as well as highlighting the need for focused genomic analysis on European regions not typically captured by large datasets to provide fine-scale insights into their genetic history and current stratification of genetic variation.SI Appendix, Supplementary Data 1.To select a European sample of genotypes from the UKBB, multiple filtering steps were carried out by firstly selecting individuals based on phenotype codes provided in the UKBB. Phenotype codes are indicated with a F.X coding, where X is a numerical code. Further data coding within each of these phenotypes are indicated with a C.X coding, where X is a numerical code. We first selected individuals with an ethnicity (F.21000) that was either “White” (C.1), “White British” (C.1001), “White Irish” (C.1002), “any other white background” (C.1003), or “other ethnic group” (C.6). The proportions of these ethnicity labels in each European country of birth group label and Leiden cluster are shown in n = 5), Azerbaijan , Georgia , and Kazakhstan ) and also excluding United Kingdom (C.354) and Irish (C.356) birthplaces. In addition to these individuals, we separately identified individuals with a British or Irish birthplace (F.1647), selecting from England (C.1), Wales (C.2), Scotland (C.3), Northern Ireland (C.4), and the Republic of Ireland (C.5). Furthermore, we identified additional individuals with a geographic birth coordinate  that corresponded to the island(s) of Orkney , Isle of Man . Lastly, we randomly down-sampled countries to 200 individuals if their sample size was larger to retain comparable sample sizes between regions.These individuals were further filtered based on a birthplace (F.20115) from Europe (C.300-C.400), excluding countries from central Asia or Caucus .This left 5,500 individuals with a common set of markers that were non-strand-ambiguous SNPs with missingness of <5%, MAF of >2%, and an HWE k ancestral components (from 2 to 7) over 10 replicate runs, choosing the replicate with the highest log-likelihood and the lowest cross-validation error.To explore the sampled population structure of Europe from 5,500 individuals from the UKBB, we initially performed PCA using PLINK , 29 and P >1e-6. We additionally pruned for linkage disequilibrium using the plink command –indep-pairwise 1000 50 0.2, leaving a final common SNP count of 46,173 variants. We performed PCA, projecting UKBB Europeans onto the genetic variation of the Human Origins West Eurasians in PLINK using the –within and –pca-cluster-names in conjunction with the –pca command.Comparing the ancestries of our European sample from the UKBB, we coanalyzed these with population references from the Human Origins dataset . We seleUsing the dataset of 5,500 Europeans and 482,591 markers, we phased haplotypes with Beagle v5.1  using deUsing the phased genotypes from Beagle, we calculated the coancestry matrix for each autosome with the pbwt  utility’With the pbwt paint chunkcount coancestry matrix as the input, we constructed a network graph using the statistical computing language R  and the fs .Visualizing haplotype sharing, we applied PCA to the pbwt paint chunk count coancestry matrix with scripts provided by the authors of fs . We addifs  implemenfs  to the tP “source” clusters, where we treated every other Leiden cluster as a potential source. For each target cluster of N individuals, we recorded a X matrix that has N rows and P columns, recording the summed chunklengths that each source cluster donates to each target cluster individual. We record a Y matrix that records the mean summed chunklength estimates between each source cluster. Using the same nnls adaption as used by Leslie et al. . This approach appears to identify the closest neighbors of haplotype identity, as well as recent admixture , and thus does not capture the deep relatedness between all Europeans  , detecti program , selectiF), namely, ROHF and SNPF  in ROH of >1.5Mb. We detected ROH using PLINK with the following parameters: –homozyg-window-snp 50; –homozyg-snp 50; –homozyg-kb 1500; –homozyg-gap 1000; –homozyg-density 50; –homozyg-window-missing 5; –homozyg-window-het 1. No linkage disequilibrium pruning was performed.SNPF is a SNP-based measure of inbreeding in the most recent generation and is an estimate of ISF (SNPF using PLINK’s –het command that is an implementation of:O(HOM) is the observed number of homozygous SNPS, E(HOM) is the expected number of homozygous SNPs given the HWE, and N is the total number of SNPs. We calculated SNPF separately for groups of clusters, as follows: NW Europe , N Britain & Ireland , England & Wales , CE Europe , E Europe , Finland 1 (Finland), Finland 2 (Mixed Scand.), S Europe , SW Europe , Mixed European (Mixed European), and Malta .e of FIS . We calcSupplementary FileSupplementary FileSupplementary FileSupplementary File


Limosilactobacillusfermentum 332 on quality characteristics in fermented sausage were explored in terms of physicochemical characteristics, volatile flavor components, and Quorum sensing (QS). The results showed that the pH of fermented sausage decreased from 5.20 to 4.54 within 24 h with the inoculation of L.fermentum 332. Lightness and redness were significantly improved, and hardness and chewiness were significantly increased after the addition of L.fermentum 332. With the inoculation of L.fermentum 332, the thiobarbituric acid reactive substance content decreased from 0.26 to 0.19 mg/100 g and total volatile basic nitrogen content decreased from 2.16 to 1.61 mg/100 g. In total, 95 and 104 types of volatile flavor components were detected in the control and fermented sausage inoculated with starter culture, respectively. The AI-2 activity of fermented sausage inoculated with L.fermentum 332 was significantly higher than that of the control and positively correlated with viable count and quality characteristics. These results provide support for further research on the effect of microorganisms on the quality of fermented food.The effects of  A great variety of fermented meats constitutes an important part of cultural patrimony. Fermented meat products have a unique flavor and texture and long shelf life under natural or artificial conditions, which are promoted by microorganisms2. Meat protein is decomposed by microorganisms and enzymes to produce a large number of amino acids, which improves the nutrition and flavor of fermented meat3. For fermented meat, four flavor development methods are used: protein degradation, lipid oxidation, the Maillard reaction, and the use of microorganisms4. However, traditional natural fermentation is difficult to normalize fermented meat quality and ensure safety. Therefore, to improve product quality and reduce production difficulties by artificial inoculation of microorganisms, which can better ensure the safety and stability of fermented meat.Fermented meats are produced worldwide due to their sensory characteristics and convenience5. Lactiplantibacillusplantarum and Streptococcusxylosus in fermented sausage promoted protein and fat decomposition while inhibiting the growth of pathogenic and spoilage bacteria to prevent odor and rancidity, maintain sausage quality, and improve flavor6. Lactobacillussake, for example, can promote lipid hydrolysis, inhibit lipid autoxidation, and improve fermented flavor development7. In terms of flavor development, LAB uses carbohydrates as the energy source to produce organic acids. Moreover, LAB can promote protein decomposition, further positively affect amino acid metabolism, and promote the development of fermented meat flavor8. The addition of LAB is beneficial to increasing the content of free fatty acids in fermented meat products, particularly to promote the release of unsaturated fatty acids and provide basic materials for flavor development10. Therefore, LAB plays an important role as a functional starter, resulting in unique flavor and safe fermented meats.Lactic acid bacteria (LAB) is a microorganism that is widely used in the production of fermented meat products. LAB in fermented meats can inhibit the growth and reproduction of pathogenic and spoilage bacteria while also lowering the content of harmful substances like nitrite11. Moreover, biofilm formation of L.sanfranciscensis12, cell adhesion of L.acidophilus13, and environmental tolerance of gram-positive bacteria14 can be controlled by QS systems. AIs are classified into several types, with AI-2 being the only signal molecule produced by both gram-negative and gram-positive bacteria and used for intraspecific or interspecific communication15. AI-2 is produced by S-ribosylhomocysteinase (LuxS), an enzyme found in many bacterial species that has been proposed to allow interspecies communication. The luxS gene has been identified in food-borne LAB, including Lactobacillus spp.16 and Bacillus spp.17, as well as food-borne pathogens18. Our previous research has shown that the LuxS/AI-2 QS system can regulate LAB metabolism and influence physiological activities20. Therefore, AI-2 might affect the quality of fermented food by regulating the metabolism of LAB. However, the QS and AI of LAB are seldom investigated in fermented foods. The knowledge of the QS of LAB during meat fermentation is rather preliminary and based on empirical observations. The relationship between AI-2 and the quality characteristics of fermented sausage has not been reported previously.Quorum sensing (QS) is a cell density-dependent type of communication mechanism mediated by QS molecules. QS molecules, also called autoinducers (AIs), enable bacteria to collectively regulate gene expression, thereby coordinating various activitiesLimosilactobacillusfermentum 332, previously isolated from fermented food with good fermentation potential for meat products and high AI-2 activity, was used as the starter for fermented sausage in this study. The changes in the activity of the signal molecule AI-2 and the quality of fermented sausage during the manufacturing process were investigated. The potential relationship between LAB AI-2 and flavors in fermented sausage was investigated preliminary, laying the theoretical groundwork for the directional improvement of fermentation strain production performance and product quality in the food industry.L.fermentum 332, was significantly higher than that in control on days 1, 5, and 11 (p < 0.05). The LAB viable count was highest during the fermentation (day 1) period, then decreased during the drying (day 5) and mature periods (day 11). As shown in Fig. L.fermentum 332 (control). The pH of control was the lowest on day 5, then slightly increased to 4.99 on day 11. During the initial stage of fermentation, optimal temperature and humidity were required for the growth of microorganisms. L.fermentum 332 was grown and metabolized rapidly under optimal conditions, producing a large amount of lactic acid and other acids. Therefore, the pH decreased rapidly, and the acidity increased. After, the microorganisms in the meat interact with the enzymes to further decompose the proteins to produce free amino acids, ammonia, and basic amines, which slightly increase the pH value22. The pH of the inoculated group was significantly lower than that of the control during the 1–11 days of fermentation (p < 0.05). These results were consistent with the results of other studies that reported the pH of the fermented sausage inoculated with LAB was lower than the pH of the control sausage28. The result showed that L.fermentum 332 has strong acidifying property, which effectively decreases the fermentation time. Meanwhile, whether the acidity change of fermented sausage caused by inoculation of L.fermentum 332 was related to the QS needed further study.The change in LAB viable count is shown in Fig. L*), redness (a*), and yellowness (b*). L* indicates the brightness of the sausage. The higher L* value indicates a brighter color of the sausage. a* indicates the bright red degree of sausage. The sausage products show attractive rose colors when the L* and a* of sausage are higher. The color changes of fermented sausage are presented in Table L* of the two groups of fermented sausage showed a downward trend. On days 1 and 5, the L* of the fermented sausage inoculated with starter culture was significantly higher than that of the control (p < 0.05). On day 11, the L* of the fermented sausage inoculated with starter culture was lower than that of the control (p < 0.05). A similar result was reported29. The addition of L.fermentum 332 reduced the aw of the fermented sausage and dried the surface of the fermented sausage. a* of fermented sausage showed an upward trend. During the mature stage (day 11), a* value of the fermented sausage inoculated with starter culture was significantly higher than that of the control (p < 0.05). No significant difference was found in b* between the two groups. b* is an important variable related to the lipid29. The results indicated that the addition of L.fermentum 332 was beneficial in improving the color of the sausage. Previous studies pointed that the LAB had the effect of improving color in fermented sausages31.Sausage color is crucial for the marketability of the meat product because it influences its appearance and acceptability. The color indexes include lightness . This may be because L.fermentum 332 increased the degradation of proteins. The reduction of sulfhydryl content caused an increase in hardness34. The result indicated that the addition of L.fermentum 332 changed the texture characteristics of the fermented sausage.Texture indices included hardness, elasticity, adhesiveness, and chewiness, all of which were affected by the meat's ripening time. For control and inoculated sausages, hardness, adhesiveness, and chewiness increased significantly with ripening time Table . The incp < 0.05). With the addition of L.fermentum 332, the TBARS decreased from 0.255 to 0.186 mg/100 g on day 11. It has been reported that the TBARS values were between 0.40 and 3.90 mg MDA/kg for vacuum-packed sausages35. The organic sausages did not exceed the value of 3.0 mg/kg, which was used as an indicator of meat oxidative rancidity36. In contrast to our findings, adding acid whey and probiotic strains to the experimental model fermented sausage had no effect (p > 0.05) on TBARS values after 0, 90, and 180 days of storage when compared to the organic sample with sea salt36. The disparity is most likely due to the different antioxidant activities of different strains.The TBARS value is the most commonly used to assess lipid oxidation in meat products. Carbonyls, aldehydes, and hydrocarbons are the main TBARS components that contribute to off-aromas and flavors in meat products. The TBARS of the two fermented sausage groups increased, as shown in Fig. L.fermentum 332 addition on TVBN in fermented sausage was shown in Fig. p < 0.05). Therefore, the L.fermentum 332 inoculation delayed the increase of TVBN and improved the quality of the fermented sausage.TVBN is the major product of protein decomposed by bacteria in meat, in which protein provides rich nutrition for microorganism growth. The effect of 37. Aldehydes, ketones, esters, acids, alcohols, and other substances would be produced during the fermentation of sausage. The quality of fermented sausage is affected by various components. The effect of adding L.fermentum 332 on volatile flavor components in fermented sausage is shown in Table 39. These compounds are typically formed as a result of protein and lipid oxidation, amino acid metabolism, and carbohydrate catabolism40. There were 95 different types of volatile flavor substances detected in the control, and 104 different types of volatile flavor substances were detected in the fermented sausage inoculated with starter culture. The proportion of esters and alcohol in the two groups' sausages was significantly higher than the proportion of other volatile flavor components.The main sources of volatile flavor components are lipid oxidative decomposition, protein degradation metabolism, and carbohydrate decompositionp < 0.05), while aldehydes, ketones, acids, olefins, and other types had relatively lower contents. Because of the addition of L.fermentum 332, the content of alcohol and ester in the fermented sausage inoculated with starter culture was significantly higher than in the control (p < 0.05). Esters are created by esterifying alcohols and acids. The majority of them have fruit and flower fragrances and contribute significantly to the flavor formation of meat products. As shown in Supplementary Tables L.fermentum 332 increased the content of ethyl caproate and ethyl octanoate in fermented sausage, and both were significantly higher than the control (p < 0.05). The alcohol types in the fermented sausage inoculated with starter culture were significantly higher than those in the control. Aldehydes are flavor compounds found in fermented sausage. N-hexanal, the basic product of linoleic acid oxidation, was found in the fermented sausage inoculated with starter culture. It has a grassy odor and reflects the degree of fat oxidation, but it was not detected in the control. Therefore, the addition of L.fermentum 332 increased the amount of flavor substances in the fermentation stage.On day 1 (fermentation stage), 62 different volatile flavor compounds were detected in the control culture and 65 different volatile flavor compounds in the fermented sausage inoculated with starter culture. Alcohols and esters had significantly higher types and contents than other types (On day 5 (drying stage), 54 types of volatile flavor compounds were detected in the control and 66 types of volatile flavor compounds in the fermented sausage inoculated with starter culture. The variety of volatile flavor compounds was lower in the drying stage than in the fermentation stage, which could be attributed to environmental changes.p < 0.05), indicating a higher degree of lipid oxidation. Esters are volatile compounds that contribute to the distinct flavor of fermented meat41. Esters accounted for 21.31% of the total volatile substances in the fermented sausage inoculated with starter culture, and the number of species was 25. The proportion of esters in total volatile substances was 18.1% in the control, and the number of species was 23. The types and contents of olefins in the fermented sausage inoculated with starter culture were greater than in the control culture (p < 0.05). Therefore, adding L.fermentum 332 improved the flavor of the fermented sausage.On day 11 (mature stage), a total of 67 types of volatile flavor compounds were detected in the control and 73 types of volatile flavor compounds in the fermented sausage inoculated with starter culture. The types and concentrations of aldehydes in the fermented sausage inoculated with starter culture were significantly higher than those in the other processing stages (L.fermentum 332 to the fermented sausage significantly decreased aw compared with that of the control (p < 0.05). aw of the inoculated group decreased from 0.830 to 0.707, and that of the control decreased from 0.832 to 0.732 after 11 days of fermentation. The differences were due to the evaporation of the surface moisture and water migration inside the sausages during the fermentation42. Our results were consistent with other reported results32. Therefore, the addition of L.fermentum 332 can effectively reduce the aw of fermented sausage, thereby extending the shelf life of the product.As shown in Fig. 43. The AI-2 activity of LAB strains, which isolated from fermented meat was detected11. Moreover, the addition of nitrate increased the luxS gene expression. The AI-2 activity of fermented sausage, which inoculated L.fermentum 332, was significantly higher than that in the control on days 1, 5, and 11 (p < 0.05). The AI-2 activity was highest in the fermentation period (day 1), then decreased during the drying period (day 5) and aging period (day 11). The change in AI-2 activity was similar to the change in LAB viable count. This indicated that the activity of AI-2 was higher in the fermented sausage inoculated with starter culture fermented sausage and consistent with the viable count of the strain. QS not only related to bacterial density but also affected by the surrounding media44.The change of AI-2 activity during the fermentation of sausage is shown in Fig. p < 0.05). This may be because the higher LAB viable count number caused the increase in AI-2 activity, which accelerated the acid production L.fermentum 332.The results showed the AI-2 activity and acid production of fermented sausage inoculated with starter culture were both significantly higher than those of the control (p < 0.05). The addition of L.fermentum 332 improved the color of the sausage over the control. This may be because L.fermentum 332 decreased the pH, and promoted nitrite to combine with myoglobin to form nitromyoglobin. Meanwhile, the hardness and chewiness of fermented sausage inoculated with starter culture were significantly higher than those of the control (p < 0.05). At this point, the fermented sausage inoculated with starter culture displayed increased AI-2 activity, which is involved in many physiological metabolic processes of LAB20. The increased AI-2 activity might promote the metabolism of L.fermentum 332, influencing the color and texture of fermented sausage.In the fermentation stage, the brightness and redness of the starter control were significantly higher than that of the control, according to the analysis of the potential correlation between AI-2 activity and color changes . This showed that adding L.fermentum 332 increased the types of volatile flavor components. Therefore, AI-2 might take part in the formation of flavor substances. However, the distinct relationship between the volatile flavor components and AI-2 activity warrants further investigation.The potential correlation between AI-2 activity and volatile flavor component changes was investigated, and it was discovered that the types of volatile flavor substances in the fermented sausage inoculated with starter culture (where AI-2 activity was higher) were significantly higher than those in the control  glycerol as frozen (80 °C) stocks. Before use, it was transferred at least three times in MRS broth  at 37 °C. Vibrioharveyi BB170 is a directionally mutated strain with an AI-2 receptor that can be used to measure AI-2 activity45. V.harveyi BB170 (ATCC BAA-1117) was cultured at 30 °C with shaking after being transferred at least three times in an autoinducer bioassay (AB) medium 46.47. The sausage's ingredients were as follows: mutton hindleg meat (70%), mutton tail fat (30%), salt (2.5%), glucose (0.5%), sugar (1%), NaNO2 (0.01%), ascorbic acid (0.05%), pepper powder (0.2%), ginger powder (0.2%), spice powder (0.1%), corn starch (1%), and lactalbumin powder (0.5%). The ingredients were thoroughly mixed and filled into pig casings. The sausage diameter was approximately 3 cm, and length was approximately 20 cm. Control was fermented sausage without starter culture and the treatment was fermented sausage inoculated with L.fermentum 332 . For 24 h, fermentation was carried out at 30 °C and 95% relative humidity (day 1). This was known as the fermentation period. For four days, the sausages were placed in a 15 °C and 75–85% relative humidity environment (day 5). This stage was regarded as the drying period. Then, the sausages were transferred to an environment of 10 °C and 65% relative humidity for 6 days (day 11). This stage was regarded as the maturation period. After preparation, the samples were packed and stored at − 20 °C until further analyses. The sausages in both groups were sampled at various fermentation times  to determine AI-2 activity, LAB viable count, physicochemical characteristics, and volatile flavor components.Fresh mutton hindleg meat and tail fat were obtained from a local commercial processor , and sausages were made with modifications to the method previously described48.Plate counts were used to determine LAB viable counts according to the method previously describedThe sausage samples were homogenized with 10 times the mass of potassium chloride solution, and the filtrate was collected to measure the pH value using a PB-10 pH meter .The sausage color was assessed using a TCP2 chromometer . The lightness (L*), redness (a*), and yellowness (b*) values of each sample were measured.3, and the texture was assessed using a QTS texture analyzer . The hardness (g), elasticity (mm), and chewiness (g) values of each sample were measured.The sausage sample was cut into 1 × 1 × 1 cmTo determine the degree of lipid oxidation, the TBARS of sausage samples was quantified. Shaking for 30 min, a 10-g minced sausage sample was mixed with 50 mL of 7.5% trichloroacetic acid (containing 0.1% ethylenediaminetetraacetic acid). Following that, 5 mL of the supernatant was filtered and mixed with 5 mL of 0.02 mol/L thiobarbituric acid solution at 90 °C for 40 min. 5 mL of chloroform was added after the mixed solution had cooled. A multifunctional microplate reader was used to measure absorbance at 532 and 600 nm . The following equation was used to calculate the TBARS value:49 with slight modifications. Of note, 5 g of the sausage sample was blended with 25 mL of distilled water and equilibrated for 30 min at room temperature. Filter paper was used to filter the solution. By adding 5 mL of 10 g/L magnesia, a 10-mL filtrate was made alkaline and distilled for 5 min. A control of 10 mL of distilled water was also used. The distillate was collected in an Erlenmeyer flask with 10 mL of 20 g/L boric acid aqueous solutions and a mixed indicator made by dissolving 0.1 g of methyl red and 0.5 g of bromocresol green into 100 mL of 95% ethanol. Titration with 0.01 mol/L hydrochloric acid solution was performed on the mixed solution. The TVBN content was calculated using the following equation:The TVBN content was determined using the method previously describedaw) was measured using an HD-3A water activity meter .Water activity , and extraction was performed for 40 min at 60 °C. After extraction, the fiber was inserted into the injection port and desorbed for 3 min at 250 °C. A gas chromatography/mass spectrometry system was used to analyze volatile compounds . The protocol was carried out exactly as previously described51. As an internal standard, 2-methyl-3-heptanone was used. Volatile compounds were identified using mass spectra obtained from the NIST MS Search 2.0 library database.Volatile flavor components were assessed using the method previously describedg for 10 min, and the pH value was adjusted to 7.0. Next, the supernatant was filtered using a bacterial filter  for sterilization. The supernatant was stored at − 80 °C until further analyses. AI-2 activity was evaluated using V.harveyi BB170 as described previously52. V.harveyi BB170 was grown in AB medium at 30 °C with shaking. The resulting cells were diluted in fresh AB medium  to OD595 nm = 0.7–1.2. Diluted V.harveyi BB170 was mixed with fermented sausage supernatant in a 100:1 (v/v) ratio. The mixture was shaken and cultured at 30 °C. The luminescence of the samples was quantified using a VICTOR X Light Luminescence Plate Reader .Minced fermented sausage and sterile distilled water were mixed at a ratio of 1:1 (w/v). The supernatant was collected after centrifugation at 12,000×p < 0.05) between the two groups of fermented sausages.All tests were repeated at least three times. Results are expressed as the mean ± standard error. Data analysis was performed using SPSS 1.0 software . A t-test was used to compare significant differences (Supplementary Table S1.Supplementary Table S2.