There is evidence that HIV prevention programs for sex workers, especially female sex workers, are cost-effective in several contexts, including many western countries, Thailand, India, the Democratic Republic of Congo, Kenya, and Zimbabwe. The evidence that sex worker HIV prevention programs work must not inspire complacency but rather a renewed effort to expand, intensify, and maximize their impact. The PLOS Collection “Focus on Delivery and Scale: Achieving HIV Impact with Sex Workers” highlights major challenges to scaling-up sex worker HIV prevention programs, noting the following: sex worker HIV prevention programs are insufficiently guided by understanding of epidemic transmission dynamics, situation analyses, and programmatic mapping; sex worker HIV and sexually transmitted infection services receive limited domestic financing in many countries; many sex worker HIV prevention programs are inadequately codified to ensure consistency and quality; and many sex worker HIV prevention programs have not evolved adequately to address informal sex workers, male and transgender sex workers, and mobile- and internet-based sex workers. Based on the wider collection of papers, this article presents three major clusters of recommendations: (i) HIV programs focused on sex workers should be prioritized, developed, and implemented based on robust evidence; (ii) national political will and increased funding are needed to increase coverage of effective sex worker HIV prevention programs in low and middle income countries; and (iii) comprehensive, integrated, and rapidly evolving HIV programs are needed to ensure equitable access to health services for individuals involved in all forms of sex work. Reflecting on the "Focus on Delivery and Scale: Achieving HIV Impact with Sex Workers" collection, David Wilson outlines action-oriented recommendations to increase the reach, intensity, and impact of HIV prevention interventions targeted towards sex work. HIV prevention programs for sex workers, especially female sex workers, are cost-effective.There are opportunities to further increase the impact of HIV prevention programs for sex workers and to adapt interventions to a changing context.Many sex worker HIV prevention programs are insufficiently guided by understanding of epidemic transmission dynamics, situation analyses, and programmatic mapping; receive limited domestic financing in many countries; are inadequately codified to ensure consistency and quality; and have not evolved adequately to address informal sex workers, male and transgender sex workers, and mobile and internet-based sex workers.We recommend increasing our understanding of HIV epidemic transmission dynamics, improving situation analyses and programmatic mapping, increasing domestic financing for sex worker HIV prevention programs where feasible, delivering well-codified, comprehensive programs using “Science of Delivery” principles and developing more effective models to reach informal sex workers, male and transgender sex workers, and mobile and internet-based sex workers.Given their marginalization, concerted efforts must be made to ensure sex workers have equitable access to HIV prevention, care, and treatment services, as well as wider health services, particularly for STIs, mental health, and addictions.The PLOS Collection “Focus on Delivery and Scale: Achieving HIV Impact with Sex Workers” amplifies evidence that HIV prevention programs targeting sex workers, particularly female sex workers, are cost-effective –20. ThesThe PLOS Collection emphasizes evidence that sex worker HIV prevention programs are robust and cost-effective. However, it must not inspire complacency. Rather, the Collection should inspire renewed determination to expand, intensify, improve the quality of, and maximize the impact of sex worker HIV prevention programs . The ColThe Collection also highlights major challenges in existing sex worker HIV prevention programs, noting the following: sex worker HIV prevention interventions are insufficiently guided by understanding of epidemic transmission dynamics, situation analyses, and programmatic mapping; sex worker HIV and STI services receive limited domestic financing in many countries; many sex worker HIV prevention programs are inadequately codified to increase consistency and quality; and many sex worker HIV prevention programs have not evolved adequately to address informal sex workers, male and transgender sex workers, and mobile- and internet-based sex workers.These findings underscore the importance of four principles of effective sex worker HIV prevention programs: (i) strengthened programmatic and contextual understanding, especially of the heterogeneity of sex work; (ii) intensified program implementation and delivery of comprehensive, proven interventions; (iii) improved program monitoring and evaluation, including greater use of real-time monitoring for continuous program improvement and constant strategic and tactical adaptations and rigorous program evaluation, to refine understanding of what works, under what circumstances and how; and (iv) predictable, sustained financing for sex worker HIV prevention programs, including increased domestic financing where feasible.The Collection also delineates the core elements of effective sex worker HIV prevention programs ,32–35, wBased on the gaps identified in the “Focus on Delivery and Scale: Achieving HIV Impact with Sex Workers” Collection and the wider literature, there are discrete clusters of action-orientated recommendations to increase the reach, intensity, and impact of HIV prevention interventions targeted towards sex work .Wherever there is sex work, there is a need to invest in programs to reduce the transmission of STIs as a core public health priority. However, the distribution, scale, and intensity of STI prevention programs targeted towards sex work must be guided by better epidemic analysis, including analysis of HIV transmission dynamics and an understanding of geographic and population heterogeneity. The overall investment in sex worker HIV prevention programs relative to total national spending on HIV prevention should be guided by the fraction of total HIV infections that are due to sex work. In some cases, sound analysis of an HIV epidemic within a country has been undertaken and has led to investment in HIV reduction programs for sex workers. For example, in India a sound epidemic analysis has led to appropriate investments in HIV reduction interventions targeted towards sex work ,36–38—InUnderstanding the contribution of sex work to a local HIV epidemic is, however, only part of the solution, as political commitment to address the health needs of sex workers is critical. In some contexts where HIV transmission sources are well characterized and sex work plays a major role, national programs do not prioritize sex worker HIV interventions . Many coSituation analyses and programmatic mapping are the foundation for high quality HIV programs. While knowing the HIV prevalence and contribution of sex work to an epidemic at a national level is important for advocacy and broad allocative funding choices, understanding the local context is critical for program management and service delivery. Situation analyses and programmatic mapping are usually the first step in an iterative process that is typically undertaken by effective HIV programs that target HIV in the context of sex work. Effective sex worker HIV prevention programs typically undertake methodical situation analyses to develop typologies of sex work, such as seaters (sex workers who operate from a fixed place) or roamers (sex workers who are mobile), or bar-based, street-based, or home-based, and to identify and map areas where sex workers live and meet clients –44. ProgThe coverage, quality, comprehensiveness, and impact of HIV prevention interventions for sex workers are variable, and consistency may be improved using implementation science –9,48–50.Many HIV prevention programs focused on sex workers have been internationally financed, including the first sex work programs in the Democratic Republic of the Congo, Kenya, Cameroon, and Zimbabwe –58. Sex The experience of providing effective health programs for sex workers is also relevant for the wider AIDS debate, without decrying the immense clinical benefits of HIV treatment and the potential for population-level HIV prevention impact, HIV treatment alone is insufficient. Access to treatment requires comprehensive services for sex workers, including actions to overcome stigma and discrimination. Treatment adherence among sex workers has powerful behavioral determinants and is reinforced by sex worker friendly services, peer support, and a supportive policy and legal context. Consistent condom use, reinforced by client programs, offers sex workers additional protection from HIV-infected clients who may or may not be on treatment and virally suppressed. Sex worker-friendly services, including STI and sexual health care, can reduce other STIs, which increase HIV susceptibility. Group solidarity and empowerment creates a powerful impetus for HIV prevention and treatment adherence and is cost-effective .The assertion that sex worker HIV prevention programs are effective applies to formal, self-acknowledged, professional sex workers. However, it is less true for informal non-self-identifying sex work and is even less true for transactional sex . While transactional sex involves the exchange of gifts, this exchange may be delayed and the link not necessarily acknowledged by either party . For thiThe HIV response must develop and evaluate intervention models for informal sex work. It must also develop and evaluate better “hotspot”  models to address a gradation of formal and informal sex work and transactional sex. This is urgently needed for several reasons: (i) as HIV epidemics mature and clients perceive formal sex workers and formal sex work venues as “higher risk,” formal networks may disperse and fragment and become harder to define and reach, a process intensified by the increasing role of mobile phones and internet in sexual solicitation ,67. ThesMobile and internet solicitation is replacing physical venues in some contexts and changing the nature of sex work as it lowers the barriers to entry into sex work and creates larger, more dispersed and fragmented and sometimes more part-time forms of sex work –71. As vMale and transgender sex workers are at great risk of stigma, discrimination, gender-based violence, and HIV infection, yet receive fewer programs and services –75. The Sex workers face a greater burden of HIV and STI infection  and a grThe “Focus on Delivery and Scale: Achieving HIV Impact with Sex Workers” Collection challenges AIDS researchers and practitioners to initiate a new generation of comprehensive sex worker HIV prevention programs for a changing sex work context and provides a framework and tools to do so, while sounding several cautions, particularly concerning informal sex workers, male and transgender sex workers, and mobile- and internet-based sex workers. It underscores the need for concerted effort to ensure marginalized communities of sex workers receive equitable access to HIV and wider health services.


Female genital cutting (FGC) is described by the World Health Organization as ‘all procedures that involve partial or total removal of the female external genitalia or other injury to the female genital organs for non-medical reasons’ . Its praWith increased societal stigma and grave misinformation, funding for FGC support structures has largely ignored sensitive issues surrounding FGC, for example, the social and economic structures that propagate abuse against women, such as poverty . WithoutFGC exemplifies a severe form of discrimination against women, stemming from historical inequalities between the sexes . AlthougDespite the obvious health implications of FGC, awareness of these issues is poor. A recent case, which clearly demonstrated the lack of awareness of FGC in British medical practice, led to the prosecution of an obstetrician in the UK . This obFGC has been criminalized in 25 African countries  – with lDespite failures in domestic policy and societal awareness, there are opportunities to advocate for FGC globally. Despite various United Nations–supported gatherings relating to FGC , this heOverall, global society has aimed to tackle FGC in multiple ways – with some gaining more successes than others. Despite the current problems revolving around poor societal awareness, ineffective legal proceedings, or poor political backing, there are many lessons we can learn to improve our support mechanisms for FGC patients worldwide. More specifically, stronger political backing, which often equates with increased financial funding, will enable us to support the broader causes of FGC by focusing on behavioural change in communities, and structural and economic barriers within society. With these structures put in place, additional health programmes and supportive policies will enable us to tackle FGC both domestically and globally – with the ultimate aim of removing FGC as a global health burden affecting the most vulnerable girls and women worldwide.Nina Al-Saadi Imperial College London London, United Kingdom Email: na2811@ic.ac.ukHarun Khan Imperial College London London, United KingdomSameera Auckburally Imperial College London London, United KingdomAdam Al-Saadi St George's University of London London, United KingdomTasnia Khan Imperial College London London, United Kingdom


The objectives of this study were to identify and assess the impact of capacity-building biosafety initiatives and programs that have taken place in the broader Middle East and North Africa (BMENA) region between 2001 and 2013, to highlight gaps that require further development, and to suggest sustainable ways to build cooperative regional biosafety opportunities.A cross-sectional study was conducted with two aspects (1) thorough desktop review of literature for all biosafety/biosecurity-related activities in the study countries, such as seminars, conferences, workshops, policy documents, technology transfer, sustained scientific endeavors between countries, etc. and (2) an online survey of scientists in countries in the region to get first-hand information about biosafety and biosecurity initiatives and gaps in their country.n = 1295) initiatives were focused on raising general awareness among the scientific community about biosafety/biosecurity/biocontainment. The most frequent areas of interest were biorisk management in biomedical and biotechnology laboratories 13% (n = 239), followed by living modified organisms (LMOs) 9.17% (n = 168). Hands-on training accounted for 2.67% (n = 49) of initiatives. Online survey results confirmed desktop review findings; however, the response rate was 11%.A total of 1832 initiatives of biosafety/biosecurity were recorded from 97 web links; 70.68% ( Recent advances in biotechnology have provided a quantum leap in the application of biological sciences in all fields, including health, agriculture, environment, and energy development. Biotechnology tools and protocols are globally available and are increasingly being used and seen as potential investments. However, with rapid advancement, widespread use, and investment, comes the possibility of exploitation of research and risk of misuse of the research outcomes for nefarious purposes. The most concerning issue remains the lack of expertise and awareness of risk management systems for the mitigation of such risks in the biological sciences, despite extensive and well-developed relevant engineering and scientific methods for containment, detection, diagnosis, treatment, and recovery as well as human behavior and performance science in other fields, such as aviation, nuclear power, petrochemicals, etc. This concern has been raised globally and has intensified since the events of 9/11 and the anthrax-in-the-mail terrorism of 2001. This has led to international focus on means for combating bioterrorism, especially in countries with the backdrop of disturbed geopolitical situations .Scientific Engagement Defining Gaps and Creating Opportunities for Cooperative Research and Global Security in the Broader Middle East and North Africa Region” to assess the overall impact of global biosecurity capacity-building initiatives undertaken in recent years in the BMENA region and to then apply the knowledge gained from this assessment to suggest sustainable ways to build cooperative regional biosafety opportunities.This study was undertaken to identify and assess the impact of capacity-building biosafety initiatives and programs that have taken place in broader Middle East and North Africa (BMENA) region between 2001 and 2013, and to highlight gaps that require further development. The project had two components: (a) a review of biosafety and biosecurity initiatives in the region and (b) assessment of the impact of these initiatives on scientists at local/regional levels. This study was part of a larger project titled “The project co-PIs, based in Pakistan and the U.S., worked closely with two Regional Coordinators in North Africa (Morocco and Egypt) to recruit a project coordinator in each of the 24 countries participating in the project. A cross-sectional descriptive study was designed consisting of two steps: (1) a desktop review/literature search for all biosafety- and biosecurity-related initiatives that were reported from the BMENA region between 2001 and 2013 using commercial search engines and (2) building a survey instrument in the commercially available SurveyMonkey (SM) platform and administering it independently to scientists in the study countries.A database was created according to the type of initiatives each study country in the region had, the scope of the activity, institute(s) involved, and type of funding/donors, etc. Great care was taken to develop a sampling process that would protect the confidentiality of survey participants and their responses. A cross-sectional online survey was conducted throughout the region. The survey questions sought to collect broad observations about overall capacity needs or gaps in biorisk management systems in the BMENA study countries. It included questions about technically skilled human resources in various aspects of biosafety management systems, availability of sustainable training programs, and challenges such as lack of human resources, monetary funds, specific bio risk management skills, etc. Questions were also developed to assess capacity for biorisk management and oversight regulation, such as availability of national/institutional biosafety committees and scientist/professionals for regulatory compliance assistance. Institutional Review Board approval was obtained through the Aga Khan University in Karachi, Pakistan. Survey subjects were identified by the country coordinators.via email.The following member countries as per World Health Organization (WHO) definition of BMENA region were included in the study: Afghanistan, Algeria, Bahrain, Egypt, Iran, Iraq, Israel, Jordan, Kuwait, Lebanon, Libya, Mauritania, Morocco, Oman, Palestine, Pakistan, Qatar, Saudi, Sudan, Syria, Tunisia, Turkey, United Arab Emirates (UAE), and Yemen. Members of biosafety associations of these countries were invited by the study country coordinators to participate in the survey. In countries without biosafety associations, members of other scientifically related associations were invited. Thirty members of these various associations from each country were identified by random selection to receive the survey questionnaire Data from desktop review were entered and analyzed using Microsoft Excel (Windows XP 2008). Results were analyzed as categorical data. The biosafety initiatives reported from member countries were categorized as per the following headings: (a) general awareness sessions: activity conducted to raise general knowledge/education of biosafety and biosecurity among the scientific community; (b) human resource development: activity that translated into hands-on training of the scientists in a particular aspect of biosafety and biosecurity with an aim to transfer technology and to provide expertise at local or regional levels; (c) institutional capacity building: activity that resulted in development of network/association/foundation that would foster the biosafety and biosecurity development at the local/regional levels; (d) scientific collaborations between the member countries or with international communities which included initiatives such as regional conferences/meetings; and (e) sustainable collaborative projects at regional levels such as infectious disease surveillance, formal laboratory biosafety and biosecurity training, responsible science/bioethics and scientific cooperation, student exchange, etc. Additional information where available was recorded regarding the duration of activity, funding support, and feedback from participants of the activity.The survey was kept open for a duration of 10 working days, and responses were directly downloaded from SM and analyzed/graphed in Microsoft Excel. The survey results were analyzed as ordinal data, responses to various survey questions were recorded as ordered categories (excellent to poor). Frequencies of the responses were generated as percentages of excellent, good, average, and poor categories of resource availability.n = 239), followed by awareness of genetically modified organisms (GMOs) 9.17% (n = 168). Very little data were available for initiatives for other areas of significant biosafety concern, such as animal health sciences, chemical industry, or radiological industry.A total of 1832 biosafety/biosecurity initiatives were recorded from 97 web links that were reviewed, after removing the duplicate, 60 web links (1–60) were used for analysis. It was encouraging to note that every country listed in the BMENA region had some initiatives related to biosafety/biosecurity between 2001 and 2013 . The topics most commonly addressed in these awareness sessions included basic biosafety, biosecurity, biocontainment, and biorisk management in biomedical and biotechnology laboratories (n = 339)  were in the category of hands-on training. Very few initiatives were identified using key words “student exchange program, regional training centers, and disease surveillance” against collective term BMENA, representing the region , the International Council for Life Scientists (ICLS), and CRDF Global. United States sponsored/funded biosafety activities organized in 2001–2013 in the BMENA region included the Department of State Cooperative Biosafety Engagement Program (CBEP) and the American Association for the Advancement of Science-Center for Science, Technology, and Security Policy (AAAS-CSTSP).n = 179), followed by Pakistan (n = 162), Morocco (n = 109), Iraq (n = 68), Iran (n = 66), and Egypt (n = 60). This involved organization and hosting of national and international conferences, seminars, and training workshops on general awareness aspects of biosafety and biosecurity . Twenty-eight recipients reviewed the survey, and 23 agreed to participate. This 11% participation rate was less than desired, although we expected a high response rate to be difficult from the countries in this study. Generally online survey responses would be expected to be in the 15–20% range, but cultural differences, possible fear of responding , and lack of experience with this type of information gathering might account for the lower rate. As a result, hard copies were posted to the country coordinators to send to their colleagues, but the response rate to the requested hard copies survey was also very poor. Iraq, Afghanistan, Sudan, Yemen, Algeria, and Syria had requested hard copies so that potential respondents would have access to all survey questions to determine if the survey was too dangerous or risky for participation. In the main release, 18% of respondents viewed the survey but did not answer the questions.Although the response rate was lower than desired, the respondents were people with relevant background. Fifty-two percent of the respondents had worked as laboratory directors/managers in their fields and 70% of these were directly involved in biorisk management activities of their institutes. The survey results are reflective of the desktop review. Seventy-one percent of the respondents marked yes to the question about initiatives on general awareness on biosafety and biosecurity in their country. Of the respondents, 52% felt that biosafety initiatives in their countries were organized by NGOs, including professional/scientific societies, while 23.8% believed they were organized by the government ministries in their country. Fifty-two percent responded in the negative to the questions asking if the international initiatives were demand driven and sustainable.excellent where the technical expertise was easily available and considered highly reliable by the local scientists; good where technical expertise was available but was not very actively sought by local scientists ; average was defined as very limited or scarce local expertise and that was not very credible ; and unavailability of any local expertise was rated as poor.Table Survey results were in accordance with the desktop review findings of biosafety initiatives during the 2001–2013 time period. We found initiatives to be largely focused on general awareness or on introductory courses on biosafety with limited human resource training and technology transfer opportunities. Consequently, there was a dearth of skilled human resources in the region as evident from survey results.Significant advancements in the field of biotechnology in the late 1990s and early 2000s raised international concerns for biosafety related to biodiversity, particularly pertaining to risk assessment and risk mitigation regarding the impacts of new research products. These concerns were more pronounced in the agriculture field with development of Bt. cotton (GMOs) and its impacts on natural plants and the environment. These international concerns, at the forefront in the United Nations Conference on Environment and Development (UNCED), were formally recognized in what became the first Earth Summit, held in Rio de Janeiro in June 1992 .Another trend noticeable from the desktop review was that countries with actively functional NGOs working as Biosafety Associations, and/or those with institutes with biosafety missions, had far more initiatives on general awareness of biosafety-related issues than those without such associations. These countries hosted biosafety programs, reflecting the heightened concern in the region. Future initiatives fostering private–public partnership are strongly recommended for successful and sustainable outcomes. Successful public/private partnership models that currently exist include the International Council for the Life Sciences (a U.S.-based non-profit agency), working in collaboration with the Royal Scientific Society of Jordan (RSS), the Biosafety and Biosecurity International Consortium , and the Moroccan Biosafety Association (MOBSA) that have developed successful sustainable projects since 2005.Conducting the online survey related to biosafety and biosecurity issues was the most complicated endeavor in this study. Obtaining statistically significant survey data in this field, and in this region, is very difficult. The participants who did respond were relevant professionals actively involved in biorisk management activities within their respective countries; thus, the responses were considered credible and are reported herein. However, the biggest challenge, and hence a limitation of this study, was the low response rate of the online survey. Regional diversity, including differences in culture, languages, dialects, and most importantly sociopolitical unrest and perception that participation in an internationally funded survey might be harmful, resulted in a sense of fear in participants despite statements declaring that all data were confidential and anonymous.Promoting trust between funders, regional scientists, and cooperative partners, and improving open communication about intentions and objectives for the bioengagement activity  in the region is of utmost importance. Bioengagement programs can greatly benefit by incorporating such efforts into mainstream national health and science programs, such as global aid programs focused on public health: malaria, soil and water parasitism, tuberculosis, vector-borne viruses, and HIV/AIDS. These programs over the years have gained the trust of scientific individuals and national governments; these programs can be used as a bridge in strengthening regional initiatives.There has been a concerted effort to enhance the general awareness of biosafety and biosecurity in the life sciences in the BMENA region over the last decade. Our study findings suggest that to date, efforts have largely been focused on raising general awareness among the broad scientific community. Also, countries with actively functional Biosafety Associations and other scientifically related associations had far more such initiatives than those without such associations, but much duplication of efforts and inefficiencies of scale have been seen over the past decade. Continuing international donor interest providing opportunities for future assistance in the development of technical expertise can lead to development of local guidelines related to issues unique to the region. Given the differences across the region, local solutions are important. Country-level analyses of the local capacity-building needs are, therefore, recommended.This study provides considerable vital information for those planning biorisk management initiatives in the region. Risk assessment and mitigation in life sciences research should be made known to a broader scientific audience, as much can be gained from similar expertise in other disciplines such as engineering, chemistry, and health physics. Efforts by donor nations and agencies to sustainably support these associations and provide biosafety trainings to their broad scientific communities may be the most efficient and effective way to build cooperative regional biosafety opportunities.EK and MC conceived the idea for this project. Literature search was conducted by NA and AH, and was reviewed and analyzed by EK, JC, AE-G, KT, and MC. The survey software was developed by HG. EK analyzed survey results and overall developed the manuscript. All authors reviewed the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Acacia thorn-prick-mediated origin of mycetoma infections, but no studies have investigated effects of soil type and Acacia geographic distribution on mycetoma case distributions. Here, we map risk of mycetoma infection across Sudan and South Sudan using ecological niche modeling (ENM). For this study, records of mycetoma cases were obtained from the scientific literature and GIDEON; Acacia records were obtained from the Global Biodiversity Information Facility. We developed ENMs based on digital GIS data layers summarizing soil characteristics, land-surface temperature, and greenness indices to provide a rich picture of environmental variation across Sudan and South Sudan. ENMs were calibrated in known endemic districts and transferred countrywide; model results suggested that risk is greatest in an east-west belt across central Sudan. Visualizing ENMs in environmental dimensions, mycetoma occurs under diverse environmental conditions. We compared niches of mycetoma and Acacia trees, and could not reject the null hypothesis of niche similarity. This study revealed contributions of different environmental factors to mycetoma infection risk, identified suitable environments and regions for transmission, signaled a potential mycetoma-Acacia association, and provided steps towards a robust risk map for the disease.In 2013, the World Health Organization (WHO) recognized mycetoma as one of the neglected tropical conditions due to the efforts of the mycetoma consortium. This same consortium formulated knowledge gaps that require further research. One of these gaps was that very few data are available on the epidemiology and transmission cycle of the causative agents. Previous work suggested a soil-borne or  Acacia distributions on mycetoma in Sudan. Here, we investigated risk factors associated with mycetoma infections in Sudan using ecological niche modeling (ENM), integrating mycetoma case records, Acacia records, and geospatial data summarizing soil, land-surface temperature, and greenness. ENMs calibrated in endemic districts were transferred across Sudan, and suggested that greatest risk was in a belt across central Sudan. Mycetoma infections occur under diverse environmental conditions; we found significant niche similarity between Acacia and mycetoma. Model predictions were amply corroborated by a preliminary assessment of a much larger mycetoma case-occurrence data base. Our results revealed contributions of different environmental factors to mycetoma risk, raised hypotheses of a causal mycetoma-Acacia association, and provide steps towards a robust predictive risk map for the disease in Sudan.WHO has recognized mycetoma as one of the neglected tropical diseases (NTDs) worldwide. Studies indicate infections from soil or possibly mediated by thorn pricks, but no detailed studies have investigated effects of soil type and  Mycetoma is a chronic, devastating, inflammatory disease of the subcutaneous tissues that spread to involve the skin, deep structures and bones, and is characterized by deformity, destruction and disability especially in late stages Acacia thorns, to the point that Acacia thorns have been found embedded in mycetoma lesions during surgery Madurella mycetomatisThe organisms causing mycetoma are geographically distributed worldwide, but are particularly common in tropical and subtropical areas, in the so-called ‘mycetoma belt,’ which includes Mexico, Venezuela, Mauritania, Senegal, Chad, Ethiopia, Sudan, Somalia, Yemen, and India M. mycetomatis DNA on Acacia thorns and in soil samples Madurella, based on the observation that M. mycetomatis is phylogentically closely related to dung-inhabiting fungi Studies to date suggest a soil-borne or thorn-prick-mediated origin of mycetoma infections Acacia-mycetoma associations based on overlap of the ecological niche of mycetoma infections with that of trees of the genus Acacia.Mycetoma ranks among the most neglected diseases worldwide, to the point that it was omitted even by major neglected tropical disease (NTD) initiatives across the globe www.ncbi.nlm.nih.gov/); we also used mycetoma data deposited in the GIDEON database (http://www.gideononline.com/). Studies were selected if they described positive mycetoma cases, and were referred to specific geographic locations that could be georeferenced precisely. When geographic references were textual in nature, latitude-longitude coordinates were assigned via reference to electronic gazetteers ; 11 records were obtained by georectification and georeferencing of et al. 2002 Occurrence records for mycetoma cases were obtained from published scientific literature via the PubMed database  to test contributions of the trees to a robust mycetoma model for Sudan and South Sudan Acacia occurrences to include only Sudan and South Sudan. All duplicate records and records lacking georeferences were excluded from analysis.Occurrence records were obtained for http://www.isric.org) to summarize chemical and physical soil characteristics (Supplementary file S-1). Soil data were obtained for each of 2 depths for each variable: 0–5 cm and 5–15 cm. Soil variables represented a collection of updatable soil property and class maps of the world at 1 km resolution produced using model-based statistical methods, including 3D regressions with splines for continuous properties and multinomial logistic regression for classes To characterize environmental variation across Sudan and South Sudan, 8-day composite Land Surface Temperature and monthly Normalized Difference Vegetation Index (NDVI) data were drawn from Moderate Resolution Imaging Spectroradiometer (MODIS) satellite imagery at 1 km spatial resolution. We also used 10 variables from the World Soil Information site (https://reverb.echo.nasa.gov/reverb/) as described in greater detail elsewhere LST and NDVI data were downloaded for 2005–2011 from the Land Processes Distributed Active Archive Center data holdings, using the NASA Reverb Echo data portal  for mycetoma and Acacia were assumed to include all of Sudan and South Sudan, based on their wide geographic distributions. We calibrated ENMs across a subset of the study region corresponding to known endemic districts; models were then transferred across all of Sudan and South Sudan for interpretation; for comparison, we also calibrated models across all of Sudan and South Sudan , although the model transfer approach should be more rigorous E = 5%) error rates The Grinnellian fundamental ecological niche is defined by the set of coarse-grained, non-interactive environmental conditions under which a species is able to maintain populations without immigrational subsidy E = 5% omission threshold.To test the ability of the ENM algorithm to predict occurrences accurately across unsampled areas of Sudan and South Sudan, we used a partial receiver operating characteristic (ROC) approach As a further, and more rigorous, test of model predictivity, we derived a preliminary view of mycetoma case data archived in the Mycetoma Research Center, in Sudan, based on cases from 1991–2014. In view of the large scale of this data resource, we selected and georeferenced ∼10% of the overall data archive at random; we eliminated cases lacking geographic references and removed records from duplicate localities, which left 158 localities for this preliminary analysis. We assessed the relationship of these data to the best of our model predictions via a one-tailed cumulative binomial probability calculation that assessed the probability of obtaining the observed level of correct prediction by chance alone, given the background expectation of correct prediction based on the proportional coverage of the region by the prediction Acacia and mycetoma. We first reassessed the accessible area (M) for both species Acacia is widely distributed and grows in a wide range of habitats M as all of Sudan and South Sudan, or alternatively as only the known mycetoma-endemic districts  had the highest partial AUC ratios, and thus appeared to perform best. Mycetoma ENM predictions indicated a band of highest environmental suitability in central Sudan between 11°S and 17°N latitude (We developed models of mycetoma cases based on (1) ENMs calibrated in endemic districts, then transferred to all of Sudan and South Sudan , and (2)latitude . Howeverlatitude : mycetomAcacia was able to reject the null hypothesis of niche similarity  of the other species. Acacia is distributed broadly across Sudan and South Sudan, whereas mycetoma infections were found only in central Sudan, but these results suggest that range difference does not reflect niche differentiation between the two .Neither of the tests comparing niches of mycetoma and P<<0.05).The coincidence between model predictions and the independent additional case data from the Mycetoma Research Center was impressive , such thKnown since the 1600s We used ENM to identify suitable sites for mycetoma infections based on environmental predictors, including dimensions thought to be associated with mycetoma cases in previous studies in Sudan Acacia thorns Acacia distributions in models improved predictions, indicating possible relevance of an Acacia-mycetoma association.Several recent studies have attempted to understand modes of entry and transmission of mycetoma Acacia may thus prove to play some role as a determinant of mycetoma distributional patterns across Sudan and South Sudan, although our results are correlational only and do not provide a direct test of this association. Our background similarity tests between ENMs for Acacia and mycetoma could not reject the hypothesis of similarity of the niches of the two species, thus at least not providing evidence against an association, and our models had greatest predictive power regarding mycetoma cases when Acacia distributions were included as environmental predictors. The important question remaining, however, is how the causative agent contacts humans, penetrates the skin, and initiates infections.Madurella mycetomatis DNA in 17 of 74 soil samples and in one of 22 thorn samples M. mycetomatis in both soil and thorn samples is of unclear importance, although perhaps culture methods are relatively insensitive or ineffective. In sum, then, our results revealed contributions of different environmental factors to mycetoma risk, identified areas suitable for mycetoma emergence, farther raised the possibility of a mycetoma-Acacia association, and provided steps towards a robust predictive risk map for the disease.Previous studies confirmed presence of Text S1The variables of the soil characteristics used in model calibration for mycetoma and Acacia spp. in Sudan. Data downloaded from the World Soil Information (http://www.isric.org). Each variable is available in 2 depths (0–5 cm and 5–15 cm).(DOC)Click here for additional data file.Text S2Potential mycetoma distribution based on occurrences across all of Sudan. These models were calibrated across all of Sudan directly based on all records collected from scientific literature and environmental variables for all of Sudan.(DOC)Click here for additional data file.


Europe's agriculture is highly specialized, dependent on external inputs and responsible for negative environmental impacts. Legume crops are grown on less than 2% of the arable land and more than 70% of the demand for protein feed supplement is imported from overseas. The integration of legumes into cropping systems has the potential to contribute to the transition to a more resource-efficient agriculture and reduce the current protein deficit. Legume crops influence the production of other crops in the rotation making it difficult to evaluate the overall agronomic effects of legumes in cropping systems. A novel assessment framework was developed and applied in five case study regions across Europe with the objective of evaluating trade-offs between economic and environmental effects of integrating legumes into cropping systems. Legumes resulted in positive and negative impacts when integrated into various cropping systems across the case studies. On average, cropping systems with legumes reduced nitrous oxide emissions by 18 and 33% and N fertilizer use by 24 and 38% in arable and forage systems, respectively, compared to systems without legumes. Nitrate leaching was similar with and without legumes in arable systems and reduced by 22% in forage systems. However, grain legumes reduced gross margins in 3 of 5 regions. Forage legumes increased gross margins in 3 of 3 regions. Among the cropping systems with legumes, systems could be identified that had both relatively high economic returns and positive environmental impacts. Thus, increasing the cultivation of legumes could lead to economic competitive cropping systems and positive environmental impacts, but achieving this aim requires the development of novel management strategies informed by the involvement of advisors and farmers. Crop production in Europe is highly specialized and currently relies on a very small number of crop species, raising questions about the sustainability of farming  nitrogen supply via biological nitrogen fixation (BNF), reducing the demand for external nitrogen fertilizers, (ii) positive pre-crop benefits through a combination of residual nitrogen and break-crop effects  is considered suitable for sandy soils with low soil pH, whereas faba bean (Vicia faba L.) is typically grown on clay soils with sufficient rainfall, soybean (Glycine max (L.) Merr.) requires appropriate daylength to flower and sufficient growing degree-days to mature, and pea (Pisum sativum L.) is suitable for many different soil types. Therefore, the design of cropping systems needs to be site-specific and based on regional expert knowledge.By introducing legumes into the cropping system, farmers also introduce more complexity into their planning. To maximize the benefits of legumes while minimizing potential threats, a number of issues need to be taken into account. The preceding crop in a rotation should have a high ability to take up available nitrogen in order to reduce residual soil N levels and allow BNF of the legumes to be optimized. Legume crops need to be kept several years apart in the rotation because of their susceptibility to soil-borne diseases. The following crop should take up the residual N of the legume quickly, in order to minimize opportunities for nitrate leaching and nitrous oxide emission. Legume crops, their management and their effects on other crops in the rotation are specific to different climatic and edaphic conditions Döring, , e.g., nAlthough the positive effects of legumes on subsequent crops, including yield benefits and reductions in nitrogen fertilizer demand, soil tillage and biocide applications are well known , (ii) calculate the impact of crop production activities , and (iii) assess and compare cropping systems with and without legumes. Following Bergez et al.  for the food market. In Västra Götaland, soils with high clay contents were selected for this study, because they represent the major cropping areas with both arable and mixed farms. Arable farmers use 80–100% of their land for cereals, and legumes and other break crops play minor roles (1.7 and 1.5% of the arable land). In forage-oriented farms, perennial grasses with a relatively low share of legumes (< 20%) are common.In Brandenburg, soil types are classified according to LELF . Arable Trifolium L.) (sole cropped or in mixture with grasses) were the most widely tested legumes. Wheat, barley and oilseed rape were the most common non-legume crops  was based on the most common crops grown.Crop rotations were generated with the previously described static and rule-based rotation generator  crop sequence suitability , (ii) maximum frequency of a crop in the rotation , (iii) minimum break between the same crop (4 years for pea), and (iv) maximum frequency of crops of the same crop type . Duplicates and multiples were removed and rotations were filtered by soil type. Agronomists checked the plausibility of the results in comparison to the existing sequences.The crop production activities (CPA) covered the crop, preceding crop and site-specific crop management. CPA were designed by collecting management data in a structured survey among 2–4 experienced agronomists in each case study in 2013. The data included inputs , outputs  and management characteristics  for each crop. In order to consider rotational effects, pre-crop types were defined according to their ability to influence the yield of the following crop through N from residue decomposition. Besides the effect on yield, differences in fertilization and agro-chemical applications were considered for each pre-crop type. The following types were separated, in the order of increasing N residue levels and positive effect on yield of subsequent crops: (i) spring cereals, winter cereals and maize; (ii) grain legumes, rapeseed and grass (< 10% legumes); (iii) forage legumes in pure stands and in mixtures with grass (>30% legumes).We selected five indicators to assess the economic and environmental performance of cropping systems. Nitrogen is not only important in crop production, it also contributes to environmental pollution of the atmosphere and water bodies, so nitrous oxide emission and nitrate leaching were selected as indicators. Nitrogen fertilizer use and nitrogen fertilizer efficiency were selected as indicators of resource use efficiency. Gross margins provided the standard indicator for economic performance of crops. All impacts were calculated on a per-hectare and per-year basis.LP is the leaching probability during the winter (mean winter precipitation divided by the water holding capacity at rooting depth and a crop-specific leaching coefficient) and the Nsurplus is calculated as:NmanureP is the plant-available N content in solid and liquid manure and Nmineralizationis calculated as a function of the total organic N content  and a region-specific mean annual mineralization rate modified by the pre-crop specific N supply level. Ndfs is the nitrogen derived from soil and is calculated as:Nitrate leaching is calculated as nitrate-N based on the soil type, preceding crop and crop management as a function of the soil leaching probability and the nitrogen surplus. Adapted from Gäth and Wohlrab  and BachNuptake is the nitrogen accumulated by the crop and Nfixation is BNF of grain and forage legumes calculated as a function of the yield, the N content, the crop-specific ratio of above and below ground plant N, the percentage of N derived from the atmosphere (%Ndfa) depending on soil mineral N content using minimum and maximum %Ndfa values  from the revenues (yield multiplied by the product price). Labor costs, subsidies, interest and insurance were not taken into account. Data for the economic analysis were obtained from regional statistics KTBL, .arable, including only grain crops, or forage, including at least one forage crop, and the groups were further divided into those with legumes or without legumes. The dataset was analyzed by plotting environmental impacts against economic impacts of all single cropping systems and by calculating the mean impact and difference between arable and forage systems with and without legumes.Cropping systems were generated for each land capability and evaluated using the impact indicators. The evaluated systems were grouped as either Trade-offs were analyzed between economic and environmental impacts using multi-criteria analysis based on five indicators: (i) Gross margin, (ii) nitrate leaching, (iii) nitrous oxide emission, (iv) N fertilizer use, and (v) N fertilizer efficiency. To allow relative comparisons, impacts were normalized for each land capability and arable and forage systems separately by dividing the impact of each single cropping system by the overall mean. Trade-offs were analyzed for selected cropping systems:current farming without legumeseconomic-environmental optimized systems without legumeseconomic-environmental optimized systems with legumesCurrent farming without legumes represents the most common cropping system per land capability based on the crop proportions and expert knowledge. Economic-environmental optimized cropping systems with and without legumes are selected from the large range of generated systems according to the criteria:Gross margin is equal to or 50€ lower than the system with the highest gross marginNitrate leaching is equal to or lower than the system with the highest gross marginNitrous oxide emission is equal to or lower than system with the highest gross marginThe selected cropping systems were compared in radar charts to identify trade-offs and to evaluate the performance from a systems perspective.Generated rotations were compared by agronomists and advisors with current and plausible cropping patterns. Algorithms for nitrate leaching were already validated by Bachinger and Zander  using HEIntegration of legumes affected the economic and environmental performance of cropping systems. The environmental performance was evaluated in terms of reduced leaching and nitrous oxide loss and lower fertilizer use, and economic performance was evaluated in terms of higher gross margin. On average across all regions, arable cropping systems with legumes had 18% less nitrous oxide emission, 24% less N fertilizer use, no increase in nitrate leaching and no decrease in gross margin  of pea and common bean in the two regions  and 9–23% lower nitrous-oxide emissions than that without legumes  win-win situations where legume cultivation is economically attractive and increases environmental benefits, or (b) situations where legumes are not economically attractive but increase environmental benefits.In the case of win-win situations, legumes are already economically attractive and provide environmental benefits. The opportunities for forage legumes are easier to utilize because of (i) their generally better economic and environmental performance compared to annual grain legumes, (ii) their high quality feed value for livestock (relatively high prices), and (iii) their relatively simple integration into existing temporary grassland. The perennial nature of forage legumes offers opportunities for climate change mitigation and adaptation  and where grain yields are relatively high. However, in most situations, grain legumes are still not competitive and there are multiple reasons why European farmers do not grow these crops. The major drivers are economic forces favoring cost-effective production systems and gains from international trade over the economic benefits of diversified production systems  design validation, (ii) plausibility checking of outputs and (iii) end-user validation, as proposed by Bockstaller and Girardin  and descThe major sensitivity of the framework was the quantification of pre-crop effects that were estimated by experts and supported by information from field experiments , and are used here to make only relative comparisons between cropping systems. Nevertheless, static approaches are often used for nitrous oxide emission  competitive but their environmental impact is considered desirable, constraints of low yield stability, weed and disease infestation, require innovative solutions from on-farm and on-station experimentation, use of crop management tools and advances in plant breeding. In such situations, policy support could be justified.All authors contributed to manuscript writing and read and approved the final manuscript. MR, GB, FS, CW, PZ, and JB designed the study. MR, PZ, and JB performed the modeling. RW, AP, IT, GB, MR, and JB provided data for the case studies and were involved in the validation of the results and interpretation.The work was financed by the German Federal Ministry of Food and Agriculture; the Brandenburg Ministry of Sciences, Research and Cultural Affairs; the Scottish Government RESAS Strategic Research Programme; the EU FP7 project “Legume Futures” (Grant 245216 CP-FP), and the FACCE-ERA-NET+ project Climate-CAFE (Grant PTJ-031A544). The publication of this article was funded by the Open Access fund of the Leibniz Association.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


To compare the proliferation and differentiation of osteoblasts derived from different locations in adolescent idiopathic scoliosis (AIS) patients.From January 2011 to May 2012, a total of 16 AIS patients  were recruited in the study, with age from 11 to 17 years. All patients received posterior spinal fusion and instrumentation, as well as thoracoplasty. Trabecular bone chips from ribs, lamina and iliac crest were harvested in each patient intraoperatively for osteoblast culture. P2 cells were used for the experiment. Proliferation and differentiation of osteoblasts were investigated by using MTT method and alkaline phosphatase (ALP) activity, respectively. One-way analysis of variance (ANOVA) was used for comparison of proliferation and differentiation among osteoblasts derived from different locations.For MTT assay, the proliferation of lamina osteoblast  was slightly lower, whereas the proliferation of rib osteoblast  was slightly higher, than that of ilium osteoblast ; however, there was no statistical difference among the 3 types of osteoblasts (P>0.05). ALP activity assay showed that the differentiation of both lamina  and rib  osteoblasts were slightly higher than that of ilium osteoblast , but without statistical significance (P>0.05).There was no difference in the proliferation and differentiation of osteoblasts derived from rib, spinal lamina and iliac crest in patients with AIS. The results of the current study indicate that local and rib bones harvested during surgery may have comparable osteogenic activities to the iliac bone grafts for spinal fusion in AIS patients, potentially eliminating the need for harvesting autologous iliac bone.


The recent era is witnessing evaluation of medicinal and nutritional value of fruits and fruit juices for the management and prevention of brain diseases like headache stress, anxiety, hypertension, and Alzheimer's and Parkinson's diseases by the scientists and researchers worldwide. Fruits possess various chemicals such as antioxidants and polyphenols, which reduce and balance the effect of hormone in brain responsible for brain disease. Natural remedy is cheap, easily available, nontoxic, and easy to prepare and provides good mental health as compared to other remedies. The main objective of this review is to acknowledge medicinal benefits of fruits for the cognition and management of brain disease.  The potential of food as prophylactic and therapeutic agent versus diseases has now begun to be acknowledged. The fascinating facts have been revealed about effect of dietary factors on certain molecular systems and mechanisms that take care of mental activity in recent years. A diet that is high in omega-3 fatty acids is earning accolades for reinforcing cognitive activity in humans  and upreNutraceutical is hybrid of nutrition and pharmaceutical and was introduced in 1989 by Stephen L. DeFelice, founder and chairperson of the Foundation of Innovation Medicine, and defined as “Food, or parts of food, that provide medical or health benefits, including the prevention and treatment of disease” , 7. FuncThere are numerous evidences regarding effect of general physical condition on children's cognitive ability and school performance , 11. TheA high risk of many mental disorders has been observed, such as attention-deficit disorder, dyslexia, dementia, depression, bipolar disorder, and schizophrenia when humans are omega-3 fatty acids deficient –22. SincFruits and fruit juice were used for cognition and brain health. Keservani and coworkers reported the use of various fruits for brain health because of medicinal value of fruits , 25. Kes Rubus genus of Rosaceae family, hybrids of these species in the Rubus subgenus, and hybrids between the Rubus and Idaeobatus subgenera provide blackberry  red and dark colored berries offer health benefits which are documented too. Nevertheless, researchers around the globe persist to express interest in this region. There has been initial research with focus on the phytochemical makeup of new and much exotic fruits and their comparison with known relatives, berries. A review written by Dr. Miller and Shukitt-Hale of the United States Department of Agriculture (USDA) human nutrition center at Tufts University proposes that intake of blueberries, strawberries, blackberries, and other berry fruits has a valuable effect on the brain and could assist in checking age-linked memory loss and other alterations. The group pleads that the berries possess neuroactive phytochemicals that act as antioxidants as well as anti-inflammatory agents .Berries are famous for their accrual of antioxidant parts  and for being the fruits offering maximum antioxidant capacity in usually consumed foods . Total p Vaccinium . Species in the section Cyanococcus are the usual fruits advertised as blueberries and are inhabitant to North America   is cultivated at large scale in Iran. This breed of orange is midseason, of red color, rich in dissolved solids, high in flavor, and seedless to some extent and bears fruits on the tree in a robust manner. The tree is strong, extremely prolific, and immune towards cold  . A commoGlobally grape  is one o Punica (its roman name) for pomegranate. The French call pomegranate as grenade whereas the Spanish utter it granada, and verbatim it means seeded (granatus) apple (pomum). The pomegranate (Punica granatum) is a deciduous shrub or stout tree, with height of 5–8 m (16 and 26 ft) [The Carthage where the finest pomegranates  were recd 26 ft) . The edid 26 ft) . The mosd 26 ft) .The neuroprotective features of pomegranate polyphenols were assessed in an animal model of Alzheimer's disease. Transgenic mice with Alzheimer's mimicking pathogenesis treated with pomegranate juice showed 50-percent fewer deposition of soluble amyloid-beta and few hippocampal amyloid accumulation than mice consuming sugar water, proposing that pomegranate juice can be neuroprotective. Animals too showed better learning of water maze accomplishments and swam faster than control animals . The preThe inhibition of serum angiotensin converting enzyme (ACE) and reduction in systolic blood pressure in hypertensive patients were observed by intake of pomegranate juice as revealed by a small clinical trial. Ten hypertensive individuals  were provided 50 mL/day pomegranate juice having 1.5 mmol total polyphenols for two weeks. Two of 7 men patients were diabetic too and two were hyperlipidemic as well. Seven of 10 subjects (70%) perceived a 36-percent mean reduction in serum ACE function and a tiny, but superb, 5-percent lowering of systolic blood pressure . The dieThe fruit and fruit juice would impart beneficial effect on brain functioning. The chemical contents have properties to alleviate the harmful effect of disease by reducing the oxidation stress or by other mechanisms. The influences of dietary factors upon neuronal process and synaptic flexibility have exhibited few of the essential underlying mechanisms for the role of diet in maintaining brain health and mental activity. The more investigations are warranted to elucidate the role of phytoconstituents in therapeutics of brain disorders.


There is growing public interest in laser therapy to treat onychomycosis, where traditional pharmaceutical options are long-term, expensive, unsuccessful, and suited to a limited demographic. Recent reviews highlighting the potential of laser therapies to offer effective, convenient, short duration treatment regimens and the need for further detailed research have not demonstrated the effectiveness of different laser types and treatment modalities. This systematic review identifies, critically appraises, synthesizes and presents the best available evidence for the effectiveness of laser treatments on onychomycosis of the nails in adults living in the community. The specific review question addressed was the following: Can laser treatment of onychomycotic nails produce outcomes comparable to the current ‘gold standard’ treatment of oral terbinafine over a minimum 12 week treatment period, for adults living in the community?This systematic review is based on the development and publication of an a priori protocol where population, intervention comparator and outcomes, and inclusion and exclusion criteria are clearly defined. A three step search strategy for published and unpublished studies in English language, in the date range 1/1/1985 to 30/6/2013 resulted in nine studies being critically appraised by two independent reviewers using the Joanna Briggs Institute Meta Analysis of Statistics, Assessment and Review Instrument (MAStARI). Seven papers were included for data extraction and synthesis. The primary outcome was cure or clinical response.There was a weak association that the neodymium-doped yttrium aluminium garnet (Nd:YAG) 1064nm laser for the treatment of onychomycosis in adults could produce clear nail growth and a mycological cure in a 12 week and 16 week period and a clinical cure at 16 weeks post treatment.These outcomes are comparable to reported results from treatment by Terbinafine. However, the evidence is tenuous at best.All the included studies reported achieving clear nail growth, and in some clinical presentations the client preference for improved nail aesthetics is paramount.Therefore the current gold standard of oral Terbinafine to achieve a cure, or Nd:YAG 1064nm laser therapy to achieve clear nail growth and improved nail aesthetics should be considered the frontline therapies, with specific choice of therapy based upon practitioner expertise and patient preference.


Hormone determination is an accurate and non-destructive method that provides demographic information concerning populations of long lived in which sex and maturity are impossible to assess by non-lethal, visual/external examination. We discuss the questions that have already been addressed with these methods, the advantages and disadvantages of them, and the demographic knowledge that can be derived for marine megafauna using hormone determination. Sex, age and sexual maturation are key biological parameters for aspects of life history and are fundamental information for assessing demographic changes and the reproductive viability and performance of natural populations under exploitation pressures or in response to environmental influences. Much of the information available on the reproductive condition, length at sexual maturity and sex determinations of endangered species has been derived from direct examination of the gonads in dead animals, either intentionally or incidentally caught, or from stranded individuals. However, morphological data, when used alone, do not provide accurate demographic information in sexually monomorphic marine vertebrate species . Hormone determination is an accurate and non-destructive method that provides indirect information about sex, reproductive condition and sexual maturity of free-ranging individuals. Correlations between sex steroid concentrations and biochemical parameters, gonadal development and state, reproductive behaviour and secondary external features have been already demonstrated in many species. Different non-lethal approaches , with intrinsic advantages and disadvantages when applied on free-ranging organisms, have been proposed to asses sex, growth and reproductive condition. Hormone determination from blood samples will generate valuable additional demographic information needed for stock assessment and biological conservation. Biological parameters describing key aspects of life history, such as sex, age structure, the dynamics of growth and sexual maturation (recruitment for breeding), are generally considered fundamental information for assessing demographic changes, reproductive viability and the performance of natural populations under exploitation pressures or in response to environmental influences, for establishing the accurate conservation status of free-ranging populations and for developing conservation and sustainable fisheries management strategies . HoweverMeasurement of hormone concentrations provides an alternative methodology that can be used for understanding physiological cycles, functional aspects of reproductive endocrinology and the relationship between the hormone dynamics and the reproductive and nesting behaviour  that are, in general, highly conserved across taxa, including elasmobranchs ; two exaIn this review, we reinforce the contribution and relevance of physiological studies as a complement to the more standard demographic data of many threatened and endangered populations of marine vertebrate species that are needed for accurate management and conservational strategies. We give particular attention to those long-lived, monomorphic marine species in which sex and maturity are impossible to assess by non-lethal, visual/external examinations. We discuss the questions that have already been addressed and the demographic knowledge derived from analytical determination of hormone concentrations in marine megafauna. We briefly review the more invasive techniques and procedures that have generated demographic data and population parameters in marine vertebrates, specifically those focused on establishing sex and reproductive status. We then summarize the information derived from each method, as well as the advantages and disadvantages of these techniques, with concluding remarks focusing on the need for wide implementation of hormone determination as a valuable, precise and non-lethal tool for those marine species for which demographic data are lacking.At present, the value  that a live free-living animal has in the wild is generally accepted ; howeverConsiderable effort in the study of reproductive endocrinology (the relationship between hormone concentrations and reproductive behaviour) has been focused on adult seabirds  have been evaluated during growth (differences between pre-maturity stages) and related to the process of sexual maturity (elevated E and T levels), providing information about the sex steroid changes related to puberty and sexual maturity. These findings complement the well-known relationship of the sex steroids with other reproductive conditions, such as the pre-ovulatory stage, mating process and spawning cycle (Carcharodon carcharias), draughtboard shark  and winter (Leucoraja ocellata) and thorny (Amblyraja radiate) skates] have emphasized that understanding the endocrine dynamics  involved during puberty and maturation would provide the opportunity to create an accurate, validated, non-lethal tool for reproductive information, such as the determination of size at sexual maturity, in free-living organisms (Sphyrna tiburo), differences in serum androgen levels are evident (P < 0.05) between stages of maturity in males ; nevertheless, quantitative correlations between the increase in hormone concentrations and gonadal maturity observed (testis size) were not evaluated (r > 0.79), the percentage of mature spermatocysts (r > 0.74) and testis mass (r > 0.79) throughout the process of sexual maturity, while female skates show a significant correlation between the E concentration and ovarian mass (r > 0.82), shell gland mass (r > 0.80) and average follicle size , green sea turtles (Chelonia mydas), Kemp's ridley sea turtles (Lepidochelys kempii) and hawksbill sea turtles (Eretmochelys imbricata), due to the lack of morphological differences between sexes at these age stages. The T and E concentrations showed significant differences between sexes in these species, facilitating the use of the E:T ratio for sex determination of the immature sea turtles  and energetic cues (e.g. body condition and lipid reserves) that stimulate gonadal development, migration, nesting behaviour and reproduction in seabirds (both sexes) and sea turtles . The E concentration is closely related to the maturation of the ovarian follicles and vitellogenesis , 2003.Pygoscelis adeliae) and Emperor penguins (Aptenodytes forsteri), the increase in plasma T concentration observed in males at the time of copulation has been related to the period of testicular development and spermatogenesis (Diomedea melanophris), grey-headed (D. chrysostoma) and wandering albatross (D. exulans), the highest levels of sex steroids in plasma can be found during the pre-laying period. In these species of seabirds, sexual differences in the dynamics of circulating P concentration can be found during the breeding cycle (In Adélie (Eudyptes chrysolophus) and Gentoo penguins (Pygoscelis papua) by variation of the plasma steroids, and concluded that delays in breeding should be related to behavioural or ecological factors instead of an immature physiological state, because late breeder male and female penguins were physiologically mature at 5 years and 1 year, respectively, prior to their first breeding event. Immature wandering albatross, which also exhibit deferred sexual maturity, have shown a significant increase in testicular size and plasma T concentration with age; even when no differences in the size of the gonads were found, immature individuals that had not yet bred, older than 5 years, showed lower concentrations of T than breeders. Albatross 10 years old were the group of immature birds (not breeders) that did not show differences in the concentration of T with respect to adults , common dolphins (Delphinus delphis) and finless porpoise (Neophocaena phocaenoides)] to assess reproductive stage (juveniles), status (pregnancy and lactation), seasonality, sex and maturity , the faecal ratio of androgen to estrogen concentration is an accurate approach for sex determination; this has been validated with photo-identified individuals  direct examination of gonads and hard body parts; (ii) mark–recapture monitoring and photo-identification; and (iii) visual examination of external morphology Table . The anaMark–recapture and sequential measurements of the same individuals at time intervals  and photThe examination of gonadal condition by histological methods and the creation of indices derived from them  have been widely used to assess the reproductive biology, reproductive condition, size at maturity and fecundity rate and for sex determination of sharks, penguins, sea turtles and marine mammals , it is possible that ultrasonography may ultimately replace the laparoscopic procedure (Laparoscopic (sea turtles and avian species) and cloacascopic examinations (penguins) are sexing techniques used in live and free-ranging immature sea turtles, chicks and adult penguins without reproductive behaviour (e.g. copulation), and have also been used to determine the reproductive condition and maturity status of sea turtles (Rhincodon typus), the sex of 85% of the individuals observed off the coast of Australia (n = 325) was identified, while diving with the animal, by clasper presence, and sexual maturity was assessed in the total of males found (100%), confirming that clasper condition criterion is a visual, non-invasive approach to assess demographic parameters for this species  and comparison of phenotypic characteristics  are non-invasive, discriminant methods to assess demographic parameters, such as reproductive stage and age/sex determination Table . In shar species . NeverthSoftness of the plaston has been suggested as a visual criterion to identify active male breeders from immature sea turtles; nevertheless, the T concentration measured in plasma has not shown a significant correlation with the percentage relative plastron softened area, and additional studies are needed . LaparosSpheniscus magellanicus) vs. bill length in adults; Despite the great sex classification power of the discriminant analysis obtained from biometric data on seabirds , laparoscopy (sea turtles), laparotomy and cloacascopy (seabirds) are the most accurate methods for determining the sex and reproductive condition of the animals, and they have been considered as the reference method (gold standard) to evaluate less-invasive approaches, such as the hormone concentrations. The main disadvantages of the classical direct approaches are logistical complications  and their invasive or lethal characteristics when applied to threatened or endangered species. Photo-identification  and biometric (seabirds), phenotypic (sharks) and behavioural data (seabirds) are non-invasive, external validation methods of sex and reproductive stages, useful only when the information is available (e.g. known size at maturity and clasper condition); they require monitoring studies of the population through the breeding season  and expensive, long time series (e.g. 25 years of life history data for cetaceans). Using phenotypic characteristics of sea turtles is difficult for immature individuals and when adults are together with sub-adults. The large discriminant accuracy of biometric data when used for sexing seabirds is limited by considerable intra-specific  and inter-specific variation.The measurement of steroid hormone concentrations has shown great accuracy as a discriminant (71–100% accuracy) of sex and reproductive status (when data could be compared), with consistent accuracy between samples (e.g. amniotic fluid and plasma) and studies . There aIn addition to the use of hormone concentrations as a sexing method, the most recent studies on sharks  and age/size at sexual maturity is centred on a few populations , habitats (nesting grounds) and reproductive stages (adults) in some species. Consequently, key demographic parameters used to establish policies and conservation strategies remain unknown. The information derived from sex steroid concentrations in different stages and size classes (


The twin forces of payors seeking fair pricing and the rising costs of developing new medicines has driven a closer relationship between pharmaceutical companies and diagnostics companies, because stratified medicines, guided by companion diagnostics, offer better commercial, as well as clinical, outcomes. Stratified medicines have created clinical success and provided rapid product approvals, particularly in oncology, and indeed have changed the dynamic between drug and diagnostic developers. The commercial payback for such partnerships offered by stratified medicines has been less well articulated, but this has shifted as the benefits in risk management, pricing and value creation for all stakeholders become clearer. In this larger healthcare setting, stratified medicine provides both physicians and patients with greater insight on the disease and provides rationale for providers to understand cost-effectiveness of treatment. This article considers how the economic value of stratified medicine relationships can be recognized and translated into better outcomes for all healthcare stakeholders.  For many years, pharmaceutical companies have struggled with declining productivity, increasing development costs and attrition of traditional blockbuster medicines. The impact being that as clinical failure rates increased, so did competition from generics and biosimilars. Healthcare authorities were also introducing increased price scrutiny and seeking ways to limit high payments for high-volume products. etc.) that is used to identify patients who will benefit by improved safety or response to a stratified medicine], can help to achieve premium pricing, improve market uptake, market size and market share in such a way that blockbuster revenues are retained [Stratified medicines  offer one potential solution to these challenges, while also offering considerable benefits in terms of patient outcomes. The use of biomarkers to identify patients in clinical studies has been shown to both reduce the size and duration of clinical studies, with considerable development cost benefits as well as a higher likelihood of success / reduced attrition rate . The traretained ,3,4,5. Iretained . Despite a wide disparity in relative markets and pricing, the relative economic contributions of the companion diagnostic test to a drug-diagnostic co-development programme, in terms of the impact to the net present value (NPV) of the associated stratified medicine is substantial . The joiSeveral articles and economic models ,11,12 haWhile it is recognized that the pharma partner (Rx) and the diagnostic partner (Dx) conduct their business on vastly different scales  with a Dx Product Profile  , with thThe economic value of the relationship will also incorporate the contributions of the diagnostic to shorter approval times, premium pricing and clearing reimbursement review hurdles in all payor settings.In Vitro Companion Diagnostic Devices” [In 2011, multiple product approvals and commercial launches provided clear examples of the benefits of companion diagnostic development and triggered updated regulatory guidance. In the US, the Food and Drug Administration (FDA) issued Draft Guidance on “Devices”  which suDevices”  and XalkDevices” . Both prIn Europe, the European Medicines Agency (EMA) released a White Paper on the use of biomarkers in drug development  and the Pricing for these targeted therapeutics has been established in context of their narrowly defined patient populations, with the strategic intent to expand markets post-approval. Xalkori  is priced at $115,200 per annum   and was Well ahead of projected view timelines, the FDA also approved Zelboraf  for the treatment of BRAF V600E mutation-positive, inoperable or metastatic melanoma, and co-approved the Cobas 4800 BRAF V600 Mutation Test, a diagnostic test developed by Roche to identify patients eligible for treatment. The pivotal phase III trial was run in a selected patient population of 675 patients with previously untreated, metastatic melanoma with the BRAF V600E mutation and showing a tremendous clinical benefit over chemotherapy .With a prevalence of BRAF mutation in melanoma of about 42% (COSMIC database ) and V60Based on the data for progression free survival from the pivotal phase III trials, a 6 months treatment period can be assumed, resulting in costs of Zelboraf treatment in the range of $56,400 ($9.400 per month) , dependiWhile the higher prices of targeted drugs may conflict with the interests of pricing agencies, there are signs that the agencies may be more amenable to a clearly defined patient population. In its rejection of Yervoy —a human monoclonal antibody for malignant melanoma approved by the FDA in March 2011 —the UK NAlthough regulatory review, pricing and payment committees may all be aware of stratified medicine, there is not yet an integrated system for the process. As an example of the number of agencies involved, we look to Germany’s approach to stratified medicines .Germany, a key pharmaceutical and diagnostic market in Europe with nearly 90% of the population being part of the public insurance system, is struggling to contain healthcare costs and to introduce new technologies clinically- and cost-effectively. A dedicated federal committee  includes representatives from payers, physicians, hospitals and wider (lay) society, and is taking public system reimbursement decisions based on input from companies, medical societies, and governmental institutions. Such government institutions include an independent organization charged with health technology assessment (HTA) called the Institute for Quality and Efficiency in Healthcare (IQWIG), founded in 2004. Based on requests from GBA, therapies as well as diagnostic tests are evaluated by IQWIG. This process may last 12 months or longer and allows companies to engage with IQWIG at certain step and results in a reimbursement recommendation to GBA. For diagnostic tests the complexity is even higher since final reimbursement may only be approved by the 16 individual German Bundesländer (regions).Whereas the demonstration of health economic benefit was only introduced to the IQWIG process in 2008, it has now become even more critical for reimbursement of therapeutics due to a recent German health reform , based oCompanion diagnostic tests on the other hand still lack such a value-based approach. As with other diagnostics, they are covered within the DRG (diagnosis related group) fee schedule in the inpatient  setting and a code-based fee schedule in the outpatient (ambulatory) setting, respectively, a technical procedural cost which does not consider clinical or economic value.As Germany is just one of the five major EU markets, the complexities of satisfying EU healthcare stakeholders are complex, evolving and ultimately multiplying. Some headway is being made in this respect through both Europe-wide harmonization of drug and diagnostic regulations, and also many global harmonization activities that include the use of companion diagnostics and stratified medicines .When personalized medicine strategies were originally developed over a decade ago, there was little commercial enthusiasm for stratifying patients for therapeutic response, but changes in the clinical and commercial environment also make a strong economic case for stratified medicine. Stratified medicines impact the economic relationship of both large pharma and companion diagnostic test developers by establishing economic risk-sharing in test development and trial operations, introducing regulatory strategy for diagnostic developers and building designated markets for final products. The joint diagnostic and drug product also creates new commercial opportunities for strategic pricing and favorable reviews by cost-effectiveness committees, both at the state and private levels. Beyond the co-developer relationship, stratified medicines impact healthcare providers at the points of testing, prescribing and payment and deliver more valuable outcomes to patients when they are receiving the appropriate care.predictive medicine, as opposed to reactive medicine, with the undoubted benefits of earlier treatment translating to better long-term outcomes. In this respect, it may well be feasible to prolong life, but the challenge of future healthcare is to ensure that quality of life remains commensurate with this longevity.Finally, more timely access to safe, highly-effective patient-stratified therapies moves healthcare towards an era of


The extent to which the latitudinal gradient in species richness may be paralleled by a similar gradient of increasing functional or phylogenetic diversity is a matter of controversy. We evaluated whether taxonomic richness (TR) is informative in terms of ecological diversity  and phylogenetic diversity (AvPD) using data on 531 mammal species representing South American old autochthonous , mid-Cenozoic immigrants  and newcomers . If closely related species are ecologically more similar than distantly related species, AvPD will be a strong predictor of ED; however, lower ED than predicted from AvPD may be due to species retaining most of their ancestral characters, suggesting niche conservatism. This pattern could occur in tropical rainforests for taxa of tropical affinity (old autochthonous and mid-Cenozoic immigrants) and in open and arid habitats for newcomers. In contrast, higher ED than expected from AvPD could occur, possibly in association with niche evolution, in arid and open habitats for taxa of tropical affinity and in forested habitats for newcomers. We found that TR was a poor predictor of ED and AvPD. After controlling for TR, there was considerable variability in the extent to which AvPD accounted for ED. Taxa of tropical affinity did not support the prediction of ED deficit within tropical rainforests, rather, they showed a mosaic of regions with an excess of ED interspersed with zones of ED deficit within the tropics; newcomers showed ED deficit in arid and open regions. Some taxa of tropical affinity showed excess of ED in tropical desert areas (hystricognaths) or temperate semideserts (xenarthrans); newcomers showed excess of ED at cold-temperate latitudes in the Northern Hemisphere. This result suggests that extreme climatic conditions at both temperate and tropical latitudes may have promoted niche evolution in mammals. During the past decade, interest in the latitudinal diversity gradient has moved away from the exclusive elucidation of species richness patterns to a more integrative consideration of phylogenetic and functional differences among species. Functional and evolutionary components of diversity offer complementary information about how species differences influence patterns of community assembly and species coexistence , 2. UntiThe latitudinal gradient in species richness may be paralleled by a similar gradient of increasing functional diversity towards the tropics if tropical species occupy more specialized niches than temperate species , 12. On At present, two hypotheses have been proposed to account for geographical variability in the association between functional and phylogenetic diversity. The tropical niche conservatism hypothesis , 20, 21 It has been suggested that tropical niche conservatism is important for mammal richness gradients ; howeverOn the other hand, niche evolution enables the invasion of new habitats and climatic regimes that had previously limited the distribution of a clade, thus reducing competition and creating additional opportunities for speciation , 27. ConIn this study we evaluated the relationships between taxonomic richness (TR), ecological diversity (ED) and phylogenetic diversity (AvPD) in mammal taxa with different histories of immigration in South America, and for which ecological data at species level were available. We recognized autochthonous xenarthrans and marsupials as living representatives of the oldest mammal lineages in South America, dating back to 65 mya , 33. MidSpatial patterns in phylogenetic diversity will parallel patterns in functional diversity if more closely related species are more similar in their ecology than less closely related species, e.g., –15. We eIf closely related species are ecologically more similar than expected based on their phylogenetic relationships, this will result in lower ED than predicted by AvPD . SeveralIf closely related species are ecologically more dissimilar than expected based on their phylogenetic relationships, this will result in higher ED than predicted by AvPD. Several factors, including environmental limitation, competition, high environmental heterogeneity and niche evolution may account for the occurrence of an excess of ED , 13.In the palaeoclimatic history of the Cenozoic in America, arid climates and open habitats arose more recently than environmental conditions similar to those of present-day tropical forests [We evaluated regional variability in the association between evolutionary and functional components of mammalian species diversity in order to address the extent to which ED and AvPD are positively associated, and to identify which regions show higher or lower ED than predicted by AvPD. We predicted three patterns of association between ED and AvPD, as follows: forests . If the  forests ), we wouCuscomys ashaninka and Salinoctomys loschalchalerosorum) for which no information of ecology or body size was available.We compiled data on the geographical distribution, functional attributes and phylogeny of 531 extant mammal species inhabiting North and South America, including marsupials , xenarthrans (N = 29), artiodactyls (N = 29), carnivorans (N = 76), hystricognaths (N = 191) and primates (N = 128). The taxonomic list and the geographical distribution of species were taken from NatureServe , excludiWe compared the list of species in NatureServe  with theWe divided the New World into equal-area grid cells of 110 km x 110 km using an equal area Mollweide projection in ArcGis 9.2 ; coastalThroughout the present study, we used the expression “ecological diversity” as an approximation to functional diversity, to mean the variety of attributes related to what organisms “do” in their habitat, so as to adopt a more flexible definition of functional diversity that does not require the association of attributes with ecosystem function . To asseData on ecological attributes were compiled to represent different kinds of resource used by species and what species do to acquire them  on the basis of 80 published sources, including books, articles and public databases . These aThis further justified taking different mammal groups separately for the analysis of diversity patterns.Missing data at species level were replaced with data of congeneric species  for each cell of the grid map using the Gower index (GI) ; the mean Gower index (ED) indicates the mean dissimilarity between pairs of coexisting species .i) for different types of attributes (i) used for each mammal group are given in Tables H-M in Before the application of the Gower index, we applied weights to different attributes to ensure that all ecological aspects contributed equally to ED calculation ; Table G(j-k) of mammal species, the Gower Index (GIjk), which ranges from 0 (complete similarity between species pairs) to 1 (complete dissimilarity between species pairs), was calculated as:For any pair ijk is the partial similarity coefficient of attribute i for the j-k pair of species.Sijk is the weight of attribute i for the j-k pair of species. Wijk = 0 if species j and k cannot be compared for attribute i because either the value of this attribute for species j (Xij) or for species k (Xik) is unknown (i.e. non-available data for any attribute in any species that is compared).WFor quantitative and ordinal attributes:s (sensu ):Sijk=0s (sensu ):ijk formula is the same as for symmetric binary attributes, but:jk for each combination of species pairs. Then we summed up the values of all combinations of species pairs and divided this number by the total number of species pairs in each cell, to produce the mean GI. The mean Gower index was our estimation of ecological diversity (ED) per cell. It has the property of being independent of the number of species being compared, so it can be used to compare values across grid cells with different species richness.Scv) as,meanGI is the mean Gower Index for each cell and GIstd is its standard deviation.We also estimated the coefficient of variation in ED  according to the mammal phylogeny of . SpeciesFor the calculation of AvPD, we adopted a pairwise distance approach similar to the estimation of Average Taxonomic Distance , adaptedTaxonomic richness (TR) was the total number of species  or genera (primates) present in each cell of the grid map.For each mammal group, the overall associations between TR, ED, and AvPD were examined using bivariate scatterplots. We used the Pearson Correlation Coefficient (r) with p-corrected for spatial autocorrelation  to evalucv and AvPD to assess geographical patterns on a continental scale. We used Geographically Weighted Regression analysis (GWR) to assess regional variability in the extent to which AvPD accounted for variation in ED, while controlling for differences in TR. To analyze geographical variability in model predictions and diversity associations, we mapped AvPD beta coefficients, local R2 and residuals obtained from GWR as applied in SAM software [2 value obtained, the better the capacity of AvPD to account for ED.We mapped TR, ED, EDsoftware , using aSafi et al.  analysedcv, AvPD and TR  in most of the mammal groups  , arid regions of northeast Brazil, northeastern coastal regions of South America , at the transition between the Neotropical and Nearctic regions, i.e., México and southern USA  and in temperate regions at high latitudes in North America (artiodactyls and carnivorans) Fig –8f. In t2 > 0.40) for some mammal groups in some arid regions  showed excess of ED for carnivorans, hystricognaths and partially also for artiodactyls Fig –10e; theArtiodactyls and carnivorans distinguished the temperate high latitudes in North America as having higher ED than predicted from AvPD and TR Fig  and 10d.Our study on South American mammals supports that species richness captures only a small amount of information about the other components of diversity , 56, 57;cv indicated where high disparity is a common phenomenon between pairs of coexisting species, and thus these areas may be of particular interest for the conservation of diversity in different mammal groups and ecosystem function: northern North America (carnivorans), tropical rainforest in the Amazon Basin (xenarthrans), the Peruvian coast (hystricognaths), and Uruguayan savannas and Caatinga .The general lack of congruence between high TR and high ED or AvPD regions reinforces the idea that different components of biological diversity must be considered in the establishment of conservation priorities , 59–61. On a continental scale, TR patterns confirmed the classical latitudinal gradient of increasing richness towards the tropics in South America; however, artiodactyls and carnivorans also showed high richness in temperate latitudes in North America, which is probably associated with the coexistence of species of different biogeographic affinity , and wiThe geographical variation in ED and AvPD showed considerable variability across different mammal groups in North and South America, which explains the weak overall statistical associations between the different components of diversity, as previously found in other studies , 63, 64.Hystricognaths, artiodactyls, and carnivorans also showed high AvPD outside the tropics. In artiodactyls and hystricognaths the occurrence of low ED and AvPD in high richness regions may reflect major radiations of only few lineages and in situ speciation, with no further successful colonization by other lineages of the same taxon . OverallOur study suggested that the general expectation of a positive relationship between ecological diversity and phylogenetic diversity might be overstated . When thTaxa of tropical affinity  highlighted the tropics as a complex mosaic of regions with excess of ED interspersed with zones of deficit of ED, thus rejecting our original prediction . This fiArtiodactyls and carnivorans are representative of newcomer lineages that migrated into South America during the GABI , 38. As The completion of the Antarctic ice sheet in Late Miocene promoted the general cooling and drying of the Patagonian region and the formation of the Argentinean pampas, in association with a major shift in vegetation . This leCtenomys species that appeared to be ecologically similar, although many aspects of their ecology are still unknown. Our results suggest that stronger selection pressures for hystricognaths might have occurred in the extreme desert regions of tropical South America than in southern South America.The general cooling and drying of southern South America was also associated with the appearance of evolutionary novelty  and theMarsupials, the remaining mammal group of tropical affinity that extend into extra-tropical latitudes, did not show an increase (or excess) in ED towards high latitudes in southern South America. Thus, our study did not support the idea of the predominance of niche evolution at high latitudes in South America. In contrast, we did find support for niche evolution in carnivorans and artiodactyls at high latitudes in North America. Interestingly, carnivorans and artiodactyls also suggested that niche evolution could possibly have been involved in the occupation of cold and humid Andean forest at high latitudes in southern South America, which suggests that cold-temperate humid forests along the southern Andes may represent “new” environmental conditions for those mammal lineages with affinity for open habitat conditions.We conclude that different components of diversity are not surrogates for one another, since a decoupling between functional and phylogenetic components of diversity might have been promoted by the occurrence of niche evolution in well-delimited regions, i.e., in association with arid environments at tropical latitudes and cold-temperate climates at extra-tropical latitudes. On the other hand, South American mammals suggest that niche conservatism is not predominant over niche evolution at tropical latitudes and thus cannot be considered the primary, single explanation to account for the latitudinal diversity gradient.S1 Dataset(XLS)Click here for additional data file.S1 File(DOC)Click here for additional data file.S2 File(DOC)Click here for additional data file.S3 File(DOC)Click here for additional data file.