Acute pain is a common reason for seeking prehospital emergency care. Regrettably, acute pain is often underestimated and poorly managed in this setting. The scoping review was conducted to gain insight into existing research on the topic and to make recommendations for future work.  To identify all available evidence related to acute pain assessment and management in the African prehospital setting, describe the extent of the evidence, encapsulate findings, and identify research gaps.  The scoping review considered primary and secondary research related to acute pain assessment and management of both medical and traumatic origins in all age groups in the African prehospital setting. The search strategy aimed to identify published, unpublished, and ongoing research which met the inclusion criteria. Potentially eligible studies were identified by a comprehensive search of electronic databases, trial registers, dissertation/thesis databases, grey literature databases, and conference proceedings. Screening and data extraction were conducted independently and in duplicate.  The comprehensive search identified 3823 potential studies, duplicate titles were removed, and 3358 titles/abstracts were screened. Full text of 66 potentially eligible titles was screened, 60 were excluded, and six publications met the inclusion criteria. Despite recommendations for pain assessment during general patient care, most studies reported no/limited pain assessment. In general, pain management was concluded to be insufficient and not conforming to best practice.  Only six publications addressing prehospital acute pain care in Africa could be identified, possibly indicative of a knowledge gap. Future research is indicated to enable a better understanding of the epidemiology of acute pain and barriers and enablers of acute pain care and to develop evidence-based clinical practice guidelines (CPGs) catering for all EMS systems in Africa. Additionally, educational initiatives should be implemented to improve the quality of acute pain care and to monitor quality through continuous quality improvement (CQI) programs. Acute pain  is a common reason for seeking emergency care (EC), in the prehospital and hospital emergency department (ED) setting. The prevalence of ED visits secondary to acute pain is between 38% and 91%  with preBeing a stressor, acute pain activates various body systems with the potential to result in numerous physiological and psychological adverse effects. If unalleviated, acute pain is associated with worse patient outcomes –18, thusAdequate pain management in the prehospital setting is both realistic and achievable, but improvement will require an understanding of the aforementioned pain management barriers and limitations in emergency medical services (EMS) systems, development of pain management policies/strategies , and invKnowledge and perceptions about pain should include the understanding that pain is influenced by various factors like culture, gender, age, language, context, previous experiences, level of consciousness, and cognition. . Pain asAlthough globally, pain and pain management are well-researched topics, given the diversity of EMS systems in Africa and the role of culture, gender, attitudes, and beliefs about pain, acute pain assessment, and management in the African prehospital setting are a pertinent area deserving in-depth exploration –30. FurtThe methodology behind scoping reviews allows for evaluating a broad research question with the intent to summerise research findings and to articulate what is known about a specific topic –38. ThisInclusion and exclusion criteria were considered in terms of type of participants, concept, context, and sources.The scoping review considered research in any age group, with patients managed by ECPs, physicians, and/or nurses in the prehospital setting, in Africa. Studies relating to neonates were excluded as it is beyond the scope of the review.The concept of interest was the assessment and management of acute pain of both traumatic and medical aetiology in the African prehospital setting.The context of the scoping review was the prehospital setting and only considered research conducted on the African continent. Prehospital refers specifically to care provided before or during transportation of the patient to hospital by EMS, and consequently, studies conducted in the aeromedical (helicopter and fixed-wing) setting and ground ambulance services were eligible. Studies related to interfacility transfers of critically ill and injured patients were excluded as pain assessment and management may be influenced by prior treatment and therefore should probably not be compared to pain care in the primary setting.The research designs considered for inclusion were primary research designs  and secondary research designs , whereas case reports, case series, and literature reviews were excluded.The search strategy aimed to identify published, unpublished, and ongoing research. Potentially eligible studies were identified by comprehensively searching the following electronic databases up to December 2018: MEDLINE, Science Direct, Scopus, Google Scholar, EBSCOhost , the Cochrane Central Register for Controlled Trials (CENTRAL), Web of Science , African Journals Online (AJOL), and Sabinet African Journals   International Clinical Trials Registry Platform were searched to identify relevant protocols, ongoing studies, and unpublished studies up to 29 November 2018. The ProQuest Dissertations and Theses database and Sabinet WorldCat Dissertations were searched for potentially relevant dissertations and theses (search up to December 2018). The grey literature database was searched for potentially relevant grey literature, and the ERIC ProQuest database was searched for potentially relevant conference abstracts or proceedings. A further effort was made in August 2018 to find grey literature by contacting emergency medicine (EM) leaders and EM societies in the African region as well as searching the following databases: OpenThesis, Networked Digital Library Theses and Dissertations, Agency for Healthcare Research and Quality, WHO: Global Index Medicus, OpenUCT, Scopus (conference proceedings), and Database of African Theses and Dissertation including Research (DATAD-R). The reference lists of included studies/thesis were reviewed for eligible publications. The corresponding authors of included studies were contacted to identify additional relevant studies .Search results were imported to the Covidence online software . A two-sData of the included studies were captured independently and in duplicate by two reviewers (AL and MM) on a data extraction form  were removed, after which 3358 titles/abstracts were screened. Sixty-six titles/abstracts were potentially eligible with 3292 records excluded. The full-text articles of the 66 potentially eligible titles were retrieved, and eligibility criteria were applied. Sixty articles/publications were excluded and six included  in the sOf the six included titles, four were peer-reviewed journal articles and one was a thesis dissertation. The sixth was grey literature published by the Professional Board for Emergency Care (PBEC), Health Professions Council of South Africa (HPCSA), and obtained through the authors' knowledge of the field of EC. One study utilised a mixed methods approach, three were observational descriptive research, one was an interrupted time series analysis, and the remaining were evidence-based CPGs. All six studies were written in English and published between 2012 and 2018. Five originated from South Africa (SA) and one from Rwanda .The key features of the six included papers are synthesised in n=57) to the survey indicated that 2% encountered <1 patient requiring analgesia per month, 28% encountered between 1 and 5 patients, 36% between 5 and 10, 19% between 10 and 15, 6% between 15 and 20, and 9% more than 20 patients per month.None of the included papers reported acute pain prevalence in the African prehospital setting. Nevertheless, in Phase 1 (quantitative phase) of the mixed methods study by Mulder , respondn=530), Matthews et al. [n=74, 14%, 95% CI: 11–17), fracture, amputations or dislocations , stabbing or gunshot wounds , chest pain , and nontraumatic pain including back pain . In four cases, diagnostic notes were not recorded. Participants (n=60) in the study by Vincent-Lambert and De Kock [In the review of PCRs  expressed that a pain score is not a good indicator for initiating analgesia. The patient's expression of comfort was deemed a good indicator for stopping analgesia, whereas the practitioner's opinion of the patient's pain in terms of the patient appearing comfortable and the patient requesting the practitioner to stop pain management was identified as factors contributing to the cessation of analgesia. Vincent-Lambert and De Kock [In phase 1 of the study by Mulder , respond De Kock  stated tUse age-appropriate pain scales as part of general patient careAll trauma patients should be considered candidates for pain reliefIn labour, meet the mother's pain relief expectationsAll patients who received analgesia must be reassessed every 5 minutes Observe patients for evidence of severe adverse effects like sedation, hypotension, hypoxia, and anaphylaxisPresence of severe adverse effects demonstrates the need to stop further administrationEC courses should teach nationally standardised age-appropriate pain scalesThe evidence-based CPGs for the South African prehospital setting by the Health Professions Council of South Africa (HPCSA)  recommenRespondents to the study by Vincent-Lambert and De Kock  indicaten=335) vs post-CQI: 92.6% (n=393); p=0.019).Limited results related to the nonpharmacological management of acute pain were obtained from the included papers. For paediatric burns victims, a Burnshield® dressing was applied by EMS in 22 (6.2%) children and 251 (71.1%) children at community health centres . HPCSA  CPGs recThe main pharmacological pain management recommendation of evidence-based CPGs by the HPCSA  is shownFor paediatric burns victims, parents and medical staff used paracetamol most frequently, whereas IV morphine in combination with oral paracetamol was administered if transported to burns units by ambulance . As evidFifty-one participants (85%) in the internet-based survey by Vincent-Lambert and De Kock  indicaten=335) vs post-CQI: 93.6% (n=393); p < 0.001) in trauma patients.Scott et al.  found thMorphine was specified as the method of analgesia in 68% of respondents in Phase 1 of Mulder's  study. FCox et al.  determinFrom the results of the scoping review, it is evident that high-quality research into prehospital acute pain assessment and management in Africa is significantly lacking. Despite extensive searches, only six papers addressing the topic could be identified. Furthermore, although Cox et al.  met the Both the WHO  and the Access to EMS in most low- and middle-income countries including the African continent is very limited –52. AccoDespite literature describing the necessity for and importance of research for the development of EMS systems in Africa , 49, resConsidering the limited number and methodological quality of the included research, this scoping review exposes the paucity of high-quality prehospital acute pain research in Africa. Except for the evidence-based CPG, no high-level evidence in the form of RCTs or systematic reviews and meta-analysis examining pain interventions in African prehospital setting could be identified. Furthermore, it is noteworthy that none of the guidelines adapted, adopted, or contextualised for the purposes of the CPGs  originatAs mentioned, none of the studies included in the scoping review investigated or reported the prevalence or any other noteworthy epidemiological characteristics of acute pain in the African prehospital setting. As previously stated, international studies indicate that acute pain in the prehospital setting is prevalent and often undertreated , 13, 15.In comparison to communicable diseases , primary healthcare (child immunization), and basic resources like running water, pain management would seem to be a low priority in the health systems of low- and middle-income countries, with a paucity of comprehensive data on pain and pain management –60.Continuous assessment of the severity of acute pain forms an integral part of acute pain management as it provides the basis for decision-making , 61; nevResearch which focuses on developing an understanding of the various challenges faced when assessing pain in the African prehospital setting is indicated. In addition, research should aim to determine pain assessment enablers and the development of pain assessment policies/strategies to guide practice and to ensure appropriate education for ECPs to facilitate effective pain assessment in the prehospital setting. Furthermore, to monitor the quality of prehospital acute pain care, EMS systems can incorporate acute pain assessment and management as clinical quality indicators and implement CQI programs to improve the quality of and accountability for prehospital pain care. The study by Scott et al.  is indicMulder , MatthewIncluded studies provided limited evidence on nonpharmacological pain management, making it a further aspect requiring additional investigation in the African prehospital setting. The literature review by Pak et al.  determinAn attempt was made to ensure that all unpublished literature on the topic of the scoping review was accessed by searching grey literature and contacting leaders in EM in Africa. Nevertheless, relevant unpublished articles or thesis dissertations may still have been missed. Despite the extended search, a very limited number of studies could be identified, and as a result, the implications for practice are limited but there are significant implications for research as the review clarifies research gaps and assists in directing focus.The majority (83%) of the studies included in the scoping review were conducted in the South African prehospital setting; this can most likely be attributed to the immaturity or lack of EMS systems in most African countries  as well A further drawback to the findings of the scoping review is that none of the included studies represented data on the patients' perspective of the quality of pain management (satisfaction), but then this may, as well, be attributed to the scarcity and immaturity of research in the African prehospital setting.Scoping review methodology does not generally require the critical appraisal of the quality of the included studies , 37, 38;Acute pain research in the African prehospital setting is significantly lacking, and large knowledge gaps exist. In order to fill the research gaps in the African prehospital setting and develop the profession, it is paramount that research capacity amongst members of the EC profession is built through education and training and that governments invest in the development of EMS systems and quality prehospital care.In terms of acute pain, it is recommended that research should focus on the following pertinent areas: gathering and publishing epidemiology data related to acute pain in the African prehospital setting, understanding providers' practice as well as barriers to and enablers of pain assessment and management in the African prehospital setting, identifying limitations within EMS systems and limitations in scope of practice, and developing evidence-based CPGs for pain assessment and management catering for all EMS systems in Africa.Due to the limited number of studies included in the scoping review, deducing implications for practice is problematic. Educational initiatives to improve the knowledge and understanding of pain assessment and management principles may prove beneficial to the quality of acute pain care. Additionally, if the scope of practice for each level of ECP qualification includes medication(s) appropriate to alleviate pain yet fitting for the level of qualification, suffering secondary to acute pain will be reduced and patient outcomes improved. Introducing pain assessment and management as EMS quality indicators will allow services to start evaluating pain care and allow for the development of CQI initiatives to advance patient care and outcomes.


Background: The observed rise in non-communicable diseases may be attributed to the ongoing changes of urban environment and society, as well as greater awareness of health-related issues and subsequent higher rates of diagnosis, which all contribute to the overall quality of life. The aim of the study was to test the association between adherence to the Mediterranean dietary pattern and self-reported quality of life in a cohort of Italian adults. Methods: The demographic and dietary characteristics of 2044 adults living in southern Italy were analyzed. Food frequency questionnaires (FFQs) and a Mediterranean diet adherence score were used to assess dietary intake. The Manchester Short Appraisal (MANSA) was used to assess self-rated quality of life. Multivariate logistic regression analyses were used to test the associations. Results: A significant linear trend of association was found for the overall quality of life and adherence to Mediterranean diet score. All of the components of the MANSA, with the exception of self-rated mental health, were individually associated with higher adherence to this dietary pattern. Conclusions: Adherence to a healthy dietary pattern is associated with the measures of better overall perceived quality of life. Modern society and “Westernized” lifestyle has been negatively associated with the mental and physical health of the population. The observed rise in non-communicable diseases may depend on ongoing changes of urban environment and society, as well as a greater awareness of health-related issues and subsequent higher rates of diagnosis . QualityThe Mediterranean diet appears to be protective against cardio-metabolic diseases ,9,10, neThe study sample was constituted of participants of the Mediterranean healthy Eating, Aging, and Lifestyles (MEAL) study, an observational investigation that is primarily focused on nutritional habits and their relation with a cluster of lifestyle behaviors that characterize the classical Mediterranean lifestyle. The study protocol with the rationale, design, and methods have been described in detail elsewhere . Briefly2), overweight (BMI 25 to 29.9 kg/m2), and obese (BMI > 30 kg/m2) [Data regarding demographic  and lifestyle characteristics  were collected. The educational level was categorized as (i) low (primary/secondary), (ii) medium (high school), and (iii) high (university), while occupational level was categorized as (i) unemployed, (ii) low (unskilled workers), (iii) medium , and (iv) high (skilled workers). Physical activity level was evaluated through the International Physical Activity Questionnaires (IPAQ) , which c0 kg/m2) . InformaLong and short food frequency questionnaires (FFQs) specifically developed and validated for the Sicilian population were used to collect dietary data ,34. The A score derived from the literature assessed the Mediterranean diet adherence: briefly, a scoring system (the MEDI-LITE score) was built while taking into consideration median (or mean) values that were reported in selected studies and two standard deviations to determine three different categories of consumption for each food group ,36. For The Manchester Short Assessment of Quality of Life (MANSA) , an instP values were based on two-sided tests and compared to a significance level of 5%. SPSS 17  software was used for all of the statistical calculations.The variables were examined for normality and skewness using the Kolmogorov–Smirnov test. The categorical variables are presented as frequencies of occurrence and percentages; differences between groups were tested using the Chi-squared test. The differences in background characteristics across participants grouped into quartiles of the MANSA score were tested. The relation between adherence to the Mediterranean diet and quality of life was tested through multivariate logistic regression analysis (when considering quartiles of the Mediterranean diet adherence score) adjusted for baseline characteristics . All of the reported In this study, we reported a direct association between the adherence to the Mediterranean diet and quality of life measured with a multi-domain instrument. However, another component of the MANSA score result was significantly associated with a higher adherence to the Mediterranean diet. The results are generally in line with previously published studies ,22,38,39High adherence to a Mediterranean dietary pattern has been associated with a longer lifespan and decreased risk of numerous non-communicable diseases, including cardio-metabolic disorders and certain cancers . PreviouFrom a mechanistic point of view, interactions between various foods and nutrients occur in the real-world nutrition and the overall variety of a diet might affect the health of individuals through a synergistic action of all its components. Antioxidant micro-nutrients and phytochemicals, such as polyphenols, which are highly contained in the Mediterranean diet, have been hypothesized to exert, at least in part, the potential beneficial effects on physical and mental health of individuals, leading to an overall better quality of life. There is evidence that the Mediterranean diet may also protect from depressive disorders and improve mental health through mechanisms that are related to inflammation, besides the aforementioned association with cardio-metabolic diseases and cancers. The most studied compounds that potentially mediate such effects are omega-3 polyunsaturated fatty acids (mainly derived from fish), monounsaturated fatty acids (mainly derived from olive oil), antioxidant vitamins including, but are not limited to, B-vitamins, vitamin D, vitamin A, and vitamin E (mainly derived from fruit and vegetable) and fiber , which have been reported to reduce the low-grade inflammatory status and improve the endothelial function and the profile of coagulation and inflammation biomarkers. Finally, results of recent research showed that a number of phytochemical compounds, such as polyphenols, have been associated with various health outcomes, such as a decreased risk of cardiovascular diseases , diabeteBesides the biological connection between the dietary patterns and health, a demonstration of the association between adherence to the Mediterranean diet and quality of life may be part of better cultural background or economic allowance that may lead to a cluster of factors that characterize a healthier lifestyle . On one The results of this study should be considered in light of some limitations. The most important issue that is common to all studies investigating quality of life, as outcome is the potential reverse causation. Indeed, the cross-sectional nature of the study does not allow for identifying a causal relationship. Among other limitations, even though a trained healthcare worker collects dietary information , data from a FFQ may be subjected to recall bias.In conclusion, higher adherence to the Mediterranean diet was associated with a higher quality of life in adults living in Southern Italy. The results from this study support further investigation that examines the relation between dietary factors and quality of life with better methodological design . Further evidence is needed to better understand the relation between such life aspects and to plan educational programs that are to improve both dietary aspects and the quality of life of the general population.


Gut microbiota and its relationship to animal health and productivity in commercial broiler chickens has been difficult to establish due to high variability between flocks, which derives from plenty of environmental, nutritional, and host factors that influence the load of commensal and pathogenic microbes surrounding birds during their growth cycle in the farms. Chicken gut microbiota plays a key role in the maintenance of intestinal health through its ability to modulate host physiological functions required to maintain intestinal homeostasis, mainly through competitive exclusion of detrimental microorganisms and pathogens, preventing colonization and therefore decreasing the expense of energy that birds normally invest in keeping the immune system active against these pathogens. Therefore, a “healthy” intestinal microbiota implies energy saving for the host which translates into an improvement in productive performance of the birds. This review compiles information about the main factors that shape the process of gut microbiota acquisition and maturation, their interactions with chicken immune homeostasis, and the outcome of these interactions on intestinal health and productivity. The demand for poultry products has grown exponentially in recent decades and it is estimated that production will reach 130 million tons of chicken meat in 2020, when it will become the most consumed animal meat in the world [The structure and functionality of the intestinal microbiota is crucial for the health of poultry since the process of acquisition and maturation of the gut microbiota throughout the growth cycle of the birds has a strong influence on the development of the intestinal epithelium and the modulation of the physiological functions required to maintain intestinal homeostasis , and in turn these functions are essential to optimize the efficiency of extraction and use of energy by the host . In thisIntestinal microbiota is considered a crucial organ that plays an integral role in maintaining the health of the host by modulating several physiological functions including nutrition, metabolism, and immunity. The digestive process is strongly linked to gut microbiota; nutrient absorption, feed digestibility, energy harvest and therefore productivity are influenced by microbiota composition and diversity ,4. The comics and multi-omics approaches as tools for the study of microbial communities has allowed a detailed characterization of the gut microbiota of chickens in a quick and robust fashion, without the need to cultivate the microorganisms that compose it, as well as describing the alterations suffered by the gut microbiota in function of different stress factors of infectious, environmental and genetic nature [The advent of c nature ,12. AlthThe initial colonization of the gastrointestinal tract of birds occurs naturally from the moment of hatching and can even begin before, by passing of microorganisms through the pores of the eggshell ,14. The There is consensus in the scientific community that early colonization of the intestine is of great importance for poultry health and productivity, since it can alter the morphology and physiology of the intestine and susceptibility to infectious diseases . The inoAfter the initial colonization of the intestine a succession of microorganisms is observed in which the species richness and complexity of the population structure of the microbiota increase as the birds grow, until eventually microbiota reaches a state of maturation and stabilizes. This process normally occurs in commercial broiler chickens around 3 weeks of life ,12,22,23The intensive breeding of chickens requires a rigorous control of the conditions in the commercial establishment to optimize the growth of the birds, mainly the temperature and the relative humidity . HoweverThere are many factors associated with the internal management and decision-making of intensive poultry production establishments that are relevant for the establishment of a healthy intestinal microbiota. The influence of some of these factors on productive performance of broiler chickens has been widely studied , but theThe litter on which the chickens are raised is usually composed of pine shavings rice hulls and other vegetal materials, and by feed, water, and chicken excreta which is mixed and composted with the bedding materials. The materials used as substrate for bedding can alter the morphology and physiology of the intestine and the composition of the intestinal microbiota . ThroughFaecalibacterium prausnitzii, a commensal butyrate-producing species, in the cecum of young chickens [Reusing litter during consecutive flocks is a practice that is usually applied to reduce production costs of aviculture in many countries. It has been reported that this practice can alter the microbiota in the litter itself, where the prevalence of halotolerant/alkaliphilic bacteria is increased, but also altered the intestinal microbiota of the birds, increasing the levels of chickens . Other achickens ,57. AlthThe intestinal health of poultry has broad implications for the systemic health of birds, animal welfare, production efficiency, food safety, and environmental impact. Although the association between animal performance and gut health is widely accepted, there is not a clear definition of gut health itself. The concept of gut health includes morphological integrity, physiological functions of the intestinal tract (including digestion and absorption of nutrients), tissue metabolism, and energy balance, developed barrier functions, efficient immune responses, sustained inflammatory balance and, particularly, an adequate microbiota. On the other hand, health can be defined as the overall condition of an animal at a given time. Diseases produce a decline in health condition resulting in poor productivity and reduced quality of the affected animals, even lead to the death.While much of the available data regarding the intestinal microbiota and enteric health relate to changes associated with a diseased state, there are clear examples of the importance of the microbiota in maintaining gut health and normal intestinal function. Several studies have focused on understanding the relationships between compositions of the native gut microbiota and colonization of the gut by pathogens. Gastrointestinal microbial community behaves as an anti-infectious barrier by inhibiting the pathogens’ adherence and subsequent colonization and the production of bacteriocins and of other toxic metabolites, excluding pathogenic microorganisms, fermenting complex polysaccharides, and providing energy to the host. Also, the initial interactions between gut microbial community and host innate immune system can lead to subsequent adaptive immune response, which can either be B-cell dependent or T-cell dependent . Many hoBeta-defensins are small antimicrobial peptides that are present on the intestinal epithelial surface and constitute an important part of innate immunity . These pGastrointestinal tract harbors diverse and complex microbial communities and, therefore, extensive and complex interactions occur between the microbial cells and host immune system. The fate of these interactions is partially determined at the very beginning development stage of chicken embryo. Early microbiota colonization is also essential for healthy gut development. Initial studies with germ-free chickens have shown that microbial exposure is necessary for an appropriate development and maturation of the gut immune system . Germ-frModification of gut microbiota in early life of chickens using oral antibiotics has been associated with subsequent immune dysfunction, including development of autoimmune diseases . ChangesCampylobacter or Salmonella [Campylobacter jejuni in chickens challenged at late days show up-regulation of IL-6, IL-17A, and IL-17F [C. jejuni alter the cytokine expression profile throughout the growth cycle and in turn this immune response can drive shifts in gut microbiota composition [Salmonella Typhimurium was described [Escherichia coli were shown to correlate with higher susceptibility to colonization by Salmonella [Lactobacillus isolates that were able to control Salmonella infection in chicken [The innate immune response leads to subsequent adaptive immune response and stimulates the production of Th2 cytokines . Increaslmonella . For exad IL-17F ,84. The position . Similarescribed . Howeverlmonella , and Lac chicken . These eSalmonella Typhimurium [E. coli, Shigella, and Salmonella are Proteobacteria with recognized pro-inflammatory mechanisms. Although no specific genera were significantly correlated with cytokine expression, the most abundant genera within phylum Proteobacteria were classified as Escherichia/Shigella [Faecalibacterium, were inversely correlated with the expression of pro-inflammatory cytokine  and positively correlated with anti-inflammatory (TGF-β4) cytokine expression, while other Firmicutes were positively correlated with the expression of pro-inflammatory cytokines [Bacteroides, differently correlated with anti-inflammatory cytokines according to the age of the birds [Not only pathogens but different commensal members of gut microbiota are able to alter the immune homeostasis. Withanage et al. reported that the expression of IL-6 increased in the cecal tonsils, ileum, and spleen on day 1 and 3 p.i. when one-day-old chickens were infected with himurium . Many GrShigella . Among Fytokines . The relhe birds .The productivity of poultry can be measured through different parameters including feed conversion ratio (FCR), residual feed intake (RFI), body weight gain (BWG), apparent metabolizable energy (AME) and time to achieve market weight . FCR is The relationship between the diversity of species in the intestinal microbiota and the feeding efficiency of poultry is not clear. In cattle, a less diverse rumen microbiota was found in more efficient animals, and the authors hypothesized that this relationship is associated with specialization of the microbiota in the production of output metabolites which are more relevant for the hosts’ energy metabolism, mainly short chain volatile fatty acids that can be used as source of energy and carbon for the growth of the animal . In lineEach section of the chicken gastrointestinal tract has a different profile of microorganism, though they are closely related. The results of microbiota studies are influenced by intrinsic characteristics of birds such as genetic, gender, age, breed, health status, as well as by farm conditions including type of diet, feed additives, environment, and farm management, among others. Lactobacillus salivarius, Lactobacillus aviarius, Lactobacillus crispatus, Clostridium lactatifermentans, different members of family Ruminococcaceae, Bacteroides vulgatus, Akkermansia, and Faecalibacterium, among others [Escherichia/Shigella were found to correlate negatively with growth and fat digestibility in broiler chickens [Campylobacter colonization of broiler chickens has been connected with reduced economic performance in terms of an increase in cumulative FCR [Lactobacillus correlated with high FCR whereas the genus Faecalibacterium correlated with low FCR. In more recent studies, restrictive feeding approach was followed to clarify the impact of feed intake on ileal and cecal microbiota and physiological and functional characteristics in broiler chickens with different residual feed intake as measure of feed efficiency [While in most trials there were shifts in alpha and beta diversity, these changes were driven by different microbial taxa. In the small intestine, previous studies have not detected differences in the microbiota of birds with high or low performance in jejunum samples ,99. In tg others ,90,93. Tchickens . Also, Ctive FCR ,100. In tive FCR . The autficiency ,97. TheyLactobacillus and Bacteroides regarding performance level [Acinetobacter, Anaerosporobacter, and Arcobacter were abundant in the lower efficiency group whereas Enterobacteriaceae and Faecalibacterium were prevalent in the better efficiency group. However, there is evidence that the relative abundance of some bacteria in feces does not correlate with the composition obtained from small or large intestines [The evidence described above involves birds sacrifice, thus the monitoring of changes in the microbiota over time implies the use of other birds, which introduces a greater variability in the determinations. Despite the advantages of cloacal swabbing as sampling method, the microbiota obtained from cloacal samples does not always reflect the diversity observed in intestines segments ,101. Stace level . Acinetotestines . TherefoGut microbiota and avian productivity are closely linked, and its association has been widely studied. However, the findings are sometimes contradictory or non-conclusive and it is difficult to identify specific bacterial populations that could reproducibly improve productivity and modulate the microbiota to a desired one, considering that the cause/effect relationship is still unclear. Further studies are needed to develop innovative tools and technologies that could contribute to enhance non-invasively gut microbiota monitoring.Gut microbiota is a complex and dynamic system with multifaceted relationships between gut microbial communities and host immune system components. It clearly must be studied as complex systems to produce robust conclusions. However, it is becoming more obvious that both microbiota and immunity are key drivers for productivity. The very first regulation of the immune system associated with constant sensing of microbes implies that indirect changes in this conditioning may have long-term consequences on the capacity of the host to mount defenses and develop inflammatory or immune conditions that would have impact in animal productivity. Alteration of the composition and function of the microbiota by diets, environmental conditions, or use of antibiotics or other strong antimicrobial compounds can transform our microbial allies into potential liabilities.The increasing use of technology and automation in chicken production establishments provides the possibility of monitoring in real time many variables in the poultry houses including physicochemical factors such as temperature, water pH, bed humidity and ammonium concentration in the air, and also bird factors such as determination of physiological parameters, imaging technologies to follow flock movements and infrared sensors to evaluate birds′ thermoregulatory features , all of


The lung offers one of the largest exchange surfaces of the individual with the elements of the environment. As a place of important interactions between self and non-self, the lung is richly endowed in various immune cells. As such, lung natural killer (NK) cells play major effector and immunoregulatory roles to ensure self-integrity. A better understanding of their abilities in health and diseases has been made possible over the past decade thanks to tremendous discoveries in humans and animals. By precisely distinguishing the different NK cell subsets and dissecting the ontogeny and differentiation of NK cells, both blood and tissue-resident NK populations now appear to be much more pleiotropic than previously thought. In light of these recent findings in healthy individuals, this review describes the different lung NK cell populations quantitatively, qualitatively, phenotypically, and functionally. Their identification, immunological diversity, and adaptive capacities are also addressed. For each of these elements, the impact of the mutual interactions of lung NK cells with environmental and microenvironmental factors are questioned in terms of functionality, competence, and adaptive capacities. As pulmonary diseases are major causes of morbidity and mortality worldwide, special attention is also given to the involvement of lung NK cells in various diseases, including infectious, inflammatory, autoimmune, and neoplastic lung diseases. In addition to providing a comprehensive overview of lung NK cell biology, this review also provides insight into the potential of NK cell immunotherapy and the development of targeted biologics. For this purpose, the airways are endowed with a broad armamentarium of cellular and humoral host defense mechanisms, most of which belong to the innate arm of the immune system. The complex interplay between resident and infiltrating immune cells acting in concert with secreted proteins, such as defensins, mucins, or collectins, shapes the outcome of host-pathogen, host-allergen, and host-particle interactions within the airway microenvironment. Among the initial checkpoints that encounter inhaled antigens and trigger pro-inflammatory or tolerogenic/anti-inflammatory downstream immune responses, natural killer (NK) cells play a key role.The lung is faced daily with 10,000 liter of inhaled air containing a myriad of particles, potentially recognized as non-self. This constant exposure of one of the most important interfaces , ILT-2, and the CD94:NKG2A heterodimer, which recognize primarily classical and non-classical major histocompatibility complex (MHC) class I molecules . NK cellIn addition to the description of this vast network of activating and inhibitory receptors, the knowledge of NK cell biology has improved in the past decades in terms of their maturation, diversity, and adaptive capacities , which aTo this end, this mini-review will focus only on certain areas, with the aim of describing the specific roles of NK cells in the lung based on the most recent and exciting advances in health and disease.+ lymphocytes . Notably, this fraction in the lung is significantly smaller than that of other tissues, such as the liver in which resident NK cells represent 50% of their total . These dNK cells .dim NK cells, considered herein as being mainly circulating lung NK cells, revealing that they disclose a terminally differentiated phenotype -γ when in contact with virally infected autologous macrophages in vitro, as compared to matched peripheral NK cells , have not yet been performed.Regarding the number of NK cells in the lung, one consideration that might be taken thus far is that the proportions of lung NK cells and their subpopulations do not appear to vary throughout life . By contIn addition to their number, the phenotype and function of NK cells could also provide information regarding their involvement in diseases. As natural cytotoxicity has been shown to be influenced by cigarette smoke, it has been also hypothesized that the functionality of lung NK cells  could bebright NK cells, disclosing an immature phenotype of NKG2A++KIRlow NK cells. Following unspecific stimulation, these lung NK cells produce a large amount of Th1 cytokines (IFN-γ and TNF-α). Whether this population belongs to resident or circulating NK cells has not yet been determined. The consequences of these observed variations have not been explored either, especially in terms of fibrosis promotion , and/or airway obstruction. Both quantitative and qualitative lung NK cell abnormalities have been described in patients with COPD , but theDeleterious involvement of stress-induced recognition could also play a role in the pathogenesis of pulmonary fibrosis: a possible predisposing factor involving the NKG2D/MICA-B pathway has been identified in patients with idiopathic pulmonary fibrosis . SimilarFurther studies are required to precisely determine the respective roles of the different lung NK cell subsets (resident vs. circulating ones) in these phenomena. Increasing our understanding of the interactions between lung NK cells and the (micro)environment , as wellImmune diversity, and NK cell diversity in particular, are essential to ensure effective recognition of the non-self. As an interface with the environment, the lung is the location of numerous infectious diseases, related to all types of pathogens. The impact of successive lung infections affecting individuals throughout life in terms of resident NK cell diversity and memory acquisition is one of the most challenging subjects of study to date. Unfortunately, no study of this kind is yet available in humans.in vitro infection  of non-small-cell lung carcinoma (NSCLC) suggested that NK cell density correlated with overall survival , 47. HowApart from these lung residency considerations, it is also important to note that modulations of educated KIRs by intra-tumoral NK cells is a key element of tumor immune surveillance. Interestingly, the exposure of NK cells to exogenous MHC-I in mice led to upregulation of the activating receptors NKp46 and NKG2D and to downregulation of Ly49C/I inhibitors  leading to a control of tumor growth . Thus, i+ NK cell progenitors reside in the bone marrow, there is a less clear understanding of the mechanisms controlling seeding of NK cells within the tissues. Whether seeding of these cells into organs generates tissue-specific NK cell maturation, or whether predefined common lymphoid progenitors with specific developmental and homing characteristics (In recent years, progress has been made in the characterization of NK cells in the lung; however, the concept of tissue resident NK cells has only recently been widely accepted, especially with the identification of residency markers, such as CD49a. These cells show important differences with the circulating NK cells in terms of phenotype and functions, which likely reflect the impact of the local micro-environment in shaping the tissue-specific characteristics of resident NK cells. The question of the ontogeny of tissue-resident NK cells remains complex and only partially explained , especiaeristics , 56 wouleristics –59 couldAll authors were involved in reading bibliography and writing the article. All co-authors reviewed the article. BH drew the 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. The reviewer FV and handling editor declared their shared affiliation at the time of review.


Trypanosoma brucei. Immune responses elicited by pathogens or systemic infections in the choroid plexus and CVOs are briefly outlined. From the choroid plexus trypanosomes can seed into the ventricles and initiate accelerated infiltration of T cells and parasites in periventricular areas. The highly motile trypanosomes may also enter the brain parenchyma from the median eminence, a CVO located at the base of the third ventricle, by crossing the border into the BBB-protected hypothalamic arcuate nuclei. A gate may, thus, be provided for trypanosomes to move into brain areas connected to networks of regulation of circadian rhythms and sleep-wakefulness, to which other CVOs are also connected. Functional imbalances in these networks characterize human African trypanosomiasis, also called sleeping sickness. They are distinct from the sickness response to bacterial infections, but can occur in common neuropsychiatric diseases. Altogether the findings lead to the question: does the neglect in reporting microbe attacks to CVOs reflect lack of awareness in investigations or of gate-opening capability by microbes?Circumventricular organs (CVOs), neural structures located around the third and fourth ventricles, harbor, similarly to the choroid plexus, vessels devoid of a blood-brain barrier (BBB). This enables them to sense immune-stimulatory molecules in the blood circulation, but may also increase chances of exposure to microbes. In spite of this, attacks to CVOs by microbes are rarely described. It is here highlighted that CVOs and choroid plexus can be infected by pathogens circulating in the bloodstream, providing a route for brain penetration, as shown by infections with the parasites  Naegleria fowleri as remarkable exceptions  do not traverse the BBB but induce inflammation in the surrounding parenchyma if trapped in vessels  through the blood, with the olfactory route used by certain amoebas such as ceptions . Invasio the BBB , 3, whil vessels –7.Another hematogenous route for parasite entry into the brain is via the choroid plexus (CP) and circumventricular organs (CVOs) though this route is largely neglected. These structures are devoid of a BBB, but a blood- cerebrospinal fluid (CSF) barrier is formed by tight junctions between the CP epithelial cells and between specialized tanycytes which outline the CVOs .Trypanosoma brucei (T. b.), the etiological agents of African trypanosomiasis, that could cause distinct brain homeostatic imbalances by their localization in the CVOs.The present overview highlights that parasites can target the CP and CVOs. The role of CP as initiator of brain infections and inflammation is briefly outlined. Focus is then on the “minute” CVOs as sites of direct parasite attacks. This is exemplified by the extracellular parasites The term CVO was coined to designate structures located around the third and fourth brain ventricles including, in mammals, the pineal gland, subcommissural organ, median eminence (ME), neurohypophysis, and organum vasculosum of the lamina terminalis (OVLT), as well as area postrema (AP) and subfornical organ (SFO) Figure  13). Th. Th13). The location of CVOs at the interface between brain tissues, bloodstream and CSF indicates a role in interactions between these compartments and in homeostatic regulations. The OVLT, AP, and SFO have sensory functions, responding to blood-borne signals, while the neurohypophysis/ME and pineal gland are secretory, releasing neurohormones into blood vessels , 21. Of The CP, where the CSF is formed, does not contain neurons and, although originally considered as a CVO , is ofteA role for the CP in initiating T cell invasion of the brain parenchyma was suggested by experimental autoimmune encephalomyelitis (EAE), a most studied paradigm of brain inflammation. In EAE, Th1 and Th17 cells can enter the brain parenchyma crossing the BBB at postcapillary venules by first traversing endothelial cells and BM to enter perivascular spaces, where antigen-presenting cells may be located , 25. T cT. b. brucei (Lymphocytic choriomeningitis virus (LCMV) is the prototype of a non-cytolytic virus that causes immune-mediated brain diseases . In adul. brucei , as descKnowledge on the activation of CVOs during systemic infections, which is raising increasing interest, has been based on the effects of the bacterial cell wall product lipopolysaccharide (LPS) . This moSigns of microglia activation in OVLT, SFO, ME, and AP are observed even in normal, unchallenged, adult mice, and this may reflect a continuous exposure to molecules from the bloodstream . InteresToxoplasma gondii tachyzoites and pseudocysts were seen in the CP epithelium and stroma in the majority of AIDS patients deceased during acute necrotizing stages of cerebral toxoplasmosis, and in 20% of the patients with healed lesions  have shown infiltration of white blood cells in the brain parenchyma, preferentially around the third ventricle and in the white matter, i.e., “leukoencephalitis”  can represent early signs of HAT  and theyT. b. infection have been documented in experimental models  . Orexinl models , 59.However, in the early stage of trypanosome infection, proinflammatory molecules from the CVOs could potentially affect the function of orexin neurons through both humoral and neural pathways. The OVLT, AP, SFO are directly connected with the LH , and oreIt is also important to note that the distributed system of regulation of sleep architecture and sleep-wake cycle has key centers in the hypothalamus and brainstem in proximity to the third and fourth ventricles , and thEarly trypanosome invasion of the Arc from the ME has been observed in infected rats . DividinT. congolense-infected goats a failure in luteinizing hormone release is due to a disruption in secretion of the gonadotropin-releasing hormone into ME vessels  synthesized by the parasites . Of thesisturbed . Interesisturbed .These data sets indicate that trypanosome attack to the CVOs elicits immune signaling mechanisms that in intensity, timing or quality differ from those of systemic bacterial infections.To our knowledge, infections of CVOs with microbes have not been reported in humans. Experimental observations on viruses are rare, but include LCMV , bat rab- Do trypanosomes have peculiar properties to promote their spread to the CVOs, or is the lack of information on direct infection of the CVOs by other microbes a consequence of lack of proper studies? Systematic neuropathological studies and application of new imaging techniques  are need- Does the targeting of trypanosomes to the CVOs elicit a spectrum of immune signals distinct in intensity or quality from those elicited by bacterial LPS in systemic infections?- Could the CVOs, defined as “windows of the brain”  implyingThe ring formed by CVOs around the third and fourth ventricles can be seen as a hub where systemic immune responses and homeostatic brain networks interact . These nAll authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.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.


Meat and meat products have been described as a very good source of angiotensin I converting enzyme (ACEI)-inhibitory peptides. The generation of bioactive peptides can occur through the action of endogenous muscular enzymes during processing, gastrointestinal digestion, or by using commercial enzymes in laboratory or industry under controlled conditions. Studies of bioavailability are necessary in order to prove the positive health effect of bioactive peptides in the body as they should resist gastrointestinal digestion, cross the intestinal barrier, and reach blood stream and target organs. However, in order to better understand their effect, interactions, and bioavailability, it is necessary to consider food matrix interactions and continue the development of quantitative methodologies in order to obtain more data that will enable advances in the field of bioactive peptides and the determination of their influence on health. Bioactive peptides derived from food proteins can exert different effects after their absorption in the human body, such as prevention of diseases or physiological modulation. Physiological properties such as antihypertensive, antioxidant, antithrombotic, or hypocholesterolemic activity in the cardiovascular system ,2; minerThe activity of angiotensin I-converting enzyme (ACEI) inhibitors has been extensively studied over the last decade. The main reason for this interest is the relevance of hypertension in the development of cardiovascular diseases, which is the most important public health problem of this century. In this respect, different synthetic drugs are available on the market for the treatment of hypertension but their numerous side effects have focused researchers’ interest on the search of alternative non-toxic and naturally generated peptides for controlling blood pressure .ACEI is a dipeptidyl carboxypeptidase enzyme that participates in the renin–angiotensin system (RAS) and converts angiotensin-I into the vasoconstrictor angiotensin-II by cleaving two amino acids at the same time, thus inactivating the vasodilator bradykinin. The role of ACEI inhibitors is to maintain the balance between the vasoconstrictive and salt-retentive effects of angiotensin-II and vasodilator effects of bradykinin . Thus, tIn this article, the generation of ACEI-inhibitory peptides from meat and meat products and their identification by empirical and in silico approaches have been reviewed, as well as the latest studies on bioavailability from the point of view of their health relevance. A discussion about current limitations and challenges to be overcome in order to advance in the state-of-the-art of this field is also included.cis/trans configurations of bonds which could influence their access/binding to the ACEI complex [ACEI-inhibitory peptides are usually small peptides with sizes comprising between 2 and 20 amino acids. Their function depends on the protein source, hydrolysis conditions, degree of hydrolysis, molecular mass, and amino acid composition as well as the position of amino acids in the peptide sequences. In this respect, ACEI-inhibitory peptides have been described that have hydrophobic and branched-chain amino acids in their structure. According to the literature, the type of amino acids located in the three positions close to the C-terminal end of the ACEI-inhibitory peptide is important for activity. The presence of aromatic, positively-charged, and basic amino acids in these positions is important for competitive binding to the ACEI active site. In fact, milk-derived tripeptides containing prolines might have different  complex ,12,13,14 complex .ACEI-inhibitory peptides obtained from food sources are inactive within the intact parent protein but can exert their activity once they are released by hydrolysis. Different ways of generating ACEI-inhibitory peptides have been utilised, as shown in 50) values of 46.56 and 256.41 µM, respectively [Gastrointestinal digestion (GI) is the last step for the generation of bioactive peptides from foods. After food ingestion, gastrointestinal peptidases such as pepsin, trypsin, or chymotrypsin are the main proteases responsible for the generation of multiple peptides, including bioactive sequences. In the laboratory, gastrointestinal digestion can be simulated using specific commercial enzymes and controlled conditions of pH and temperature. Thus, a simulated gastrointestinal digestion of raw pork meat using pepsin and pancreatin indicated that the physiological digestion of pork proteins could generate peptides with biological activity . These rectively , using tectively . Later, ectively . A recenectively .On the other hand, GI simulation has also been used in studies of bioavailability of certain peptide sequences in order to demonstrate whether they could exert a positive health effect in the body, as they should resist further GI digestion, cross the intestinal barrier, and reach the blood stream and target organs.Finally, necessary treatments of meat before consumption such as cooking could facilitate the later generation of bioactive peptides due to denatured proteins being more susceptible to be hydrolysed by the enzymes of the intestinal tract.Aspergillus oryzae, and Neutrase and Alcalase from Bacillus subtilis and Bacillus lincheniformis, respectively, have been the most used in the generation of bioactive peptides. In addition, proteases from plant origin such as bromelain and papain have been described as interesting enzymes for the hydrolysis of meat proteins by their contribution to meat tenderisation. These enzymes show a wider specificity in comparison with other enzymes such as trypsin or pepsin, cleaving peptide bonds from a wide variety of regions and frequently acting as either endopeptidases, or as exopeptidases hydrolysing amino acids from N- and C-terminal sites. In fact, the activity and hydrolytic specificity of many commercial peptidases is not clearly defined by manufacturers and thus, the degree of hydrolysis and final content of peptides is difficult to predict [The most used methodology for the generation of bioactive peptides is the hydrolysis of proteins with commercial enzymes. Proteases from different sources such as of microbial, plant, or animal origin, have been used for the hydrolysis of food proteins. In meat and meat products, Flavourzyme from  predict .Several studies have reported the generation and identification of ACEI-inhibitory peptides resulting from hydrolysates of pork ,21,22, cBioactive peptides can be generated through the action of endogenous enzymes in ageing and curing processes as well as in combination with microbial peptidases such as in fermentation processes. Proteolysis by endogenous proteases is the most important phenomena occurring in the ageing of meat that influences its final characteristics with endogenous peptidases as main figures. Broadly speaking, endopeptidases such as calpains and cathepsins are first responsible for the hydrolysis of proteins into large fragments and oligopeptides, which affect the texture of meat during ageing and the initial steps of curing processes. Later, the activity of exopeptidases such as aminopeptidases and carboxypeptidases will generate small peptides and free amino acids, responsible for the characteristic flavour of dry-cured products. Some of the generated small peptides have also been described as bioactive peptides, exerting activities such as ACEI-inhibitory activity and antihypertensive, antioxidant, antilisteria, dipeptidyl peptidase IV (DPP-IV) inhibitory, and anti-inflammatory activity.Lactobacillus sakei, Lactobacillus curvatus, Lactobacillus plantarum and Lactobacillus casei alone or in combination with staphylococci, Kocuria, yeast, or moulds, exert proteolysis through the action of endo- and exopeptidases. In general, these fermentation processes are involved in the liberation of small peptides and free amino acids that not only affect flavour development but also contribute to the generation of bioactive peptides [Dry-fermented sausages are elaborated using shorter processes with microorganisms such as lactic acid bacteria (LAB) (as a starter), yeasts, or moulds that are responsible for fermentation followed by ripening/drying. Lactic acid bacteria such as peptides .The presence of ACEI-inhibitory peptides naturally generated during the processing of meat products such as dry-cured hams or dry-fermented sausages has also been described ,31,32,33Traditionally, empirical approaches have been the method of choice for the identification of bioactive peptides from food matrices. However, it is very challenging when the objective is to generate specific peptide sequences that are able to exert certain activity. Then, the experimental design can be simplified by using bioinformatics for computer simulation in silico.Empirical approaches used for the identification of bioactive peptides including ACEI-inhibitory peptides in complex sample matrices such as meat and meat products involve: (1) the release of the bioactive sequences from the parent protein; (2) preliminary in vitro assays to screen for bioactivity; (3) purification and separation through the use of high-resolution techniques, such as chromatography; (4) additional in vitro assays to determine the most active fractions; (5) identification of peptides by mass spectrometry (MS) techniques; (6) selection and synthesis of potential bioactive peptides; and (7) in vitro and in vivo confirmation of the bioactivity . A schemo-aminobenzoylglycyl-p-nitrophenylalanylproline. However, the inhibitory activities of these peptides on ACEI activity do not always correlate with antihypertensive effects. In this regard, SHRs are the animal model most frequently used to verify the in vivo efficacy of ACEI-inhibitory peptides. Some studies have evaluated the effects on SBP of SHRs after oral administration of meat hydrolysates or peptide extracts showing ACEI-inhibitory activity [In vitro ACEI-inhibitory activity is typically measured by monitoring the conversion of a specific substrate by ACEI in the presence and absence of inhibitors. Spectrophotometric and chromatographic methods have been commonly used to measure the hydrolysis of substrates such as Hippuryl-His-Leu (HHL) or the fluorogenic activity ,35,36,37activity ,42,43,44activity , whereasactivity . Additioactivity ,48,49.In addition to empirical approaches, the use of in silico analyses that combine bioinformatics tools and peptide databases has been increasingly used as a cost- and time-effective alternative. This predictive strategy enables to obtain biological and chemometric information on peptide sequences to be obtained through a series of steps: (1) selection of proteins of interest with known amino acid sequences by predicting their potential as precursors of novel bioactive peptides; (2) in silico protein digestion by selected proteolytic enzymes; (3) in silico identification and characterisation of peptides; (4) bioactivity prediction using a combination of sequence biochemical properties and databases of known bioactive peptides; (5) peptide synthesis; and (6) in vitro or in vivo confirmation of the bioactivity . Figure In silico approaches have shown that some bovine, porcine and chicken proteins such as collagen, connectin, and myosin are good sources of ACEI-inhibitory peptides, which could be released from the parent protein through the action of determined enzymes ,53,54. MAt the end, empirical and in silico approaches converge in the need for the confirmation of both the identity of the generated/predicted peptide sequences and their activity in the complexity of the matrix see .Bioavailability studies are necessary to assess whether the bioactive peptide can reach its target site in active form and sufficient quantity to exert health effects in the organism. The action of gastrointestinal enzymes, intestinal absorption, cellular uptake, and action of blood plasma peptidases can modify the structure of ACEI-inhibitory peptides or hydrolyse them leading to a loss, maintenance, or gain of bioactivity ,56.The use of digestion models and LC-MS techniques combined with in silico and in vitro/in vivo approaches have enabled evaluation of the stability of ACEI-inhibitory peptides from beef, chicken and pork meat in gastrointestinal digestion as well as the identification and quantification of the resulting products ,57,58,59The bioavailability and bioaccessibility of bioactive peptides can also be affected by processing/storage conditions and food matrix-peptide interactions that can lead to peptide modifications with changes in its native structure and activity . SeveralCurrently, with the basis of knowledge for the identification of bioactive peptides already clear, it is necessary to continue the research in bioactive peptides to achieve a better understanding of their effect, interactions, and bioavailability. In this sense, several authors have established the need for serious consideration of food matrix interactions, especially when the objective is to use the bioactive peptides as a functional ingredient . IncreasOn the other hand, increased effort on the development of quantitative methodologies for a better understanding of hydrolysis, bioactivity, and/or bioavailability is necessary. Data such as the quantity of specific naturally generated peptides in the original food and the dose of a bioactive peptide needed to exert an effect in vivo, as well as the final sequences and amount present in bloodstream and target organ after GI digestion are key data for advancing in the field of bioactive peptides and their health influence. In fact, determining the quantity of ACEI-inhibitory peptides in the meat sample that are able to reach the target site in the human system is of fundamental importance in bioavailability studies to better understand the effects and mechanisms of action of these peptides. The main limitation for quantitation is the nature of sample: small peptides often comprise fewer than four amino acids at low abundance, and there is high complexity of the matrix . CurrentMeat and meat products have been described as a very good source of ACEI-inhibitory peptides. With proteins being a major constituent of meats, the generation of bioactive peptides from meat proteins has been described as occurring either through the action of endogenous muscular enzymes during processing, during GI digestion, or by using commercial enzymes in the laboratory or in industrial processes under controlled conditions. The identification of ACEI-inhibitory peptides has been traditionally done using empirical approaches, although currently there is an increasing interest in in silico approaches based on bioinformatics as they are less time-consuming and cheaper methodologies. However, despite the identification of bioactive peptides being clear, there is an increasing need to study food matrix interactions, especially when the objective is to use the bioactive peptides as a functional ingredient. The quantitation of these peptides for a better understanding of their health influence and bioavailability is necessary to advance in this field.


Behavioural activation (BA) aims to increase positive response-contingent environmental reinforcement and help caregivers to engage in pleasant and constructive activities, and therefore improve psychological and physical health among family caregivers of people with dementia (PWD). However, knowledge of the effectiveness of BA in this population remains limited. The current study applied a systematic review and meta-analysis in order to determine the effectiveness of BA among family caregivers of PWD. Literature was searched in PubMed, Medline, CINAHL, Cochrane, Embase and PsycINFO published from March 1988 to March 2018. Seven Randomized Control Trials (RCT)s evaluating the effects of BA in family caregivers of PWD were eligible to be included in this review. Cochrane’s guideline was used in order to measure risk of bias and extract data. A random effects model was used to pool the effect size. Family caregivers of PWD receiving BA that only for caregivers demonstrated a statistically significant reduction in depression . BA also had a positive impact on interlukin-6, negative affect of caregiving, relationship satisfaction, dysfunctional thoughts and distress related to neuropsychiatric symptoms of PWD for family caregivers. The available evidence suggests that future studies are needed to focus on better ways of administering BA to family caregivers of PWD, to improve their physical and psychological health. Meanwhile, more RCTs to investigate the effects of BA on psychological and physical health for family caregivers of PWD is needed.


The software was optimized using synthetic datasets to represent the great diversity of virus genomes. The application was then validated with next generation sequencing data of hundreds of viruses. User time is minimal and it is limited to the time required to upload the data.Genome Detective is an easy to use web-based software application that assembles the genomes of viruses quickly and accurately. The application uses a novel alignment method that constructs genomes by reference-based linking of http://www.genomedetective.com/app/typingtool/virus/.Available online: Bioinformatics online. In the rapidly expanding field of genomics, our ability to produce data far exceeds our capacity to analyze and extract meaningful information. Analysis of viral data is particularly challenging given the high variability of viruses and their deviation from reference genomes, the increasing speed of identification, the continuous emergence of new viruses and the relative scarcity of viral fragments in metagenomic samples  data and/or high-throughput Sanger data. In response to this need, we have developed Genome Detective, a web-based bioinformatics pipeline to accurately and quickly identify, assemble and classify all known viruses present in NGS and Sanger sequencing data.Genome Detective accepts unprocessed paired-end or single reads generated by NGS platforms in FASTQ format and/or processed FASTA sequences. For FASTQ files, low-quality reads are filtered and adapters trimmed with Trimmomatic  of the hits identified by DIAMOND. However, some reads that represented the same viral species were assigned to buckets at different taxonomic ranks. We solved this problem by either distributing the reads from the node downwards, or collapsing them upwards, by comparing the number of reads identified at each node of the taxonomy tree versus in all descendant nodes. In addition, given that metagenomic studies are accelerating  , which ide novo consensus sequence by selecting the detailed report  graphics are available for viral species, genome images, alignment viewer, nt and amino acid similarity measures and read counts. In addition, the user can produce a bam file with BWA  using thd report  and acceWe first validated Genome Detective using a synthetic virus dataset (NCBI SRA: SRR3458562-SRR3458569), originally prepared to optimize laboratory-based virus extraction procedures, in which viruses were carefully selected to cover the range of naturally occurring diversity . These iWe compared our assignment results with IVA , which ide novo contigs. Despite the use of only RefSeq for the identification of virus species, sensitivity and specificity were maintained due to the use of both nt and amino acid similarity scores. We found that for large NGS and metagenomic datasets, Virus Detective substantially reduces computational cost without compromising the quality of the result. However, the construction of de novo whole genomes from metagenomic samples depends on the number of reads, the virus genome size and read length. Our pipeline also allows detailed displays of data and results. Furthermore, Genome Detective is linked to our popular virus-specific typing tools , a Royal Society Newton Advanced Fellowship (TdO), the VIROGENESIS project receives funding from the European Union’s Horizon 2020 Research and Innovation Program (under Grant Agreement no. 634650) and the National Institutes of Health Common Fund, grant number U24HG006941. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. We would like to acknowledge the contribution of Annelies Kroneman, Harry Vennema, Roel Standaert, Pieter Libin and Kristof Theys.Conflict of Interest: none declared.Supplementary InformationClick here for additional data file.Supplementary TableClick here for additional data file.


The fate of PCDDs, PCDFs, and PCBs in sewage sludges after different management techniques — such as agricultural application, land restoration, and marine disposal — was studied. Changes observed in the concentrations, in the ratio between PCDD and PCDF levels, and in the isomeric distribution suggest the influence of the sewage sludge on the sludge-treated areas (soils and sediments). Whereas land application techniques seem to produce no serious environmental consequences, marine disposal practices produce considerable increases in the levels of contamination in marine sediments.


Chronic obstructive lung disease (COLD) is a group of airway diseases, previously known as emphysema and chronic bronchitis. The heterogeneity of COLD does not allow early diagnosis and leads to increased morbidity and mortality. The increasing number of COLD incidences stresses the need for precision medicine approaches that are specific to the patient. Metabolomics is an emerging technology that allows for the discrimination of metabolic changes in the cell as a result of environmental factors and specific genetic background. Thus, quantification of metabolites in human biofluids can provide insights into the metabolic state of the individual in real time and unravel the presence of, or predisposition to, a disease. In this article, the advantages of and potential barriers to putting metabolomics into clinical practice for COLD are discussed. Today, metabolomics is mostly lab-based, and research studies with novel COLD-specific biomarkers are continuously being published. Several obstacles in the research and the market field hamper the translation of these data into clinical practice. However, technological and computational advances will facilitate the clinical interpretation of data and provide healthcare professionals with the tools to prevent, diagnose, and treat COLD with precision in the coming decades. Haemophilus influenzae, Streptococcus pneumoniae and sometimes Staphylococcus aureus that is difficult to treat [Chronic Obstructive Lung Disease (COLD) develops as a chronic and persistent inflammatory response to environmental stimuli. It is characterized by airflow limitation and affects the 10%–15% of the adult population globally, smoking being the most important risk factor for this disease [to treat ,3,4. Morto treat .COLD is a very heterogeneous disease and therefore requires advanced and specialized methods for diagnosis . TraditiSeveral metabolomic studies in recent years have aimed to identify and validate specific biomarkers that would help in prognosis, as well as facilitating the early and precise diagnosis of the disease, considering the great heterogeneity of the pathogenesis of COLD . The ideThere are two main analytical strategies used in metabolomics studies for the identification and evaluation of biomarkers, the untargeted and the targeted metabolomics analysis. The main differences between the two strategies are the level of sample preparation required, the number of the metabolites detected, and the level of the quantification of the metabolites. The selection of the appropriate strategy in each case is mostly based on these objectives . More spOn the other hand, targeted metabolomics aims to analyze specific metabolites and fully quantify their changes. It analyses a group of a small number of metabolites that are chemically characterized, predetermined, and involved in known metabolic pathways, providing their accurate identification and quantification. In targeted metabolomics, only the metabolites of interest are retained during sample preparation, while the rest of the biological species are removed. During the statistical analysis, itis determined how well the selected metabolites contribute to the separation of the groups under investigation, e.g., between the control and the phenotype of interest. As expected, data analysis and biological interpretation is simpler in studies that use targeted metabolomics compared to those using untargeted metabolomics. Moreover, targeted methods have a greater selectivity and sensitivity than untargeted methods, as they can be performed only if a standard curve of the metabolite is available. Finally, they perform full and accurate quantification of the selected metabolites by using internal and chemical standards for each of the metabolite in the study .Most of the metabolomic data obtained the past few years on COLD were based on untargeted metabolomics studies aimed at the identification of unknown metabolites, serving as biomarkers. In particular, untargeted metabolomic studies sought to identify specific biomarkers for COLD by measuring differences in metabolites between healthy individuals and individuals with COLD. More specifically, several studies showed that aminoacid metabolism, lipid metabolism, energy metabolism, and oxidative stress pathways were disturbed in COLD patients compared to healthy controls. One of the biggest clinical studies, with respect to the number of individuals involved, included a control group of 66 healthy individuals and three groups of 70, 64 and 44 Global Initiative for Chronic Obstructive Lung Disease (GOLD) patients with different stages of COLD  . In thisSimilarly, in 2018, studies using HPLC-MS showed altered aminoacid metabolism, including decreased levels of the aminoacids arginine, proline, alanine, and phenylalanine. Moreover, the free carnitine to acylcarnitine ratio which is important for energy metabolism was decreased, while lysophosphatidylcholine, which is involved in lipid metabolism, was increased in the serum of COLD patients compared with healthy individuals ,17. ConsCOLD is in many cases affected by environmental and genetic factors, a fact that makes the discovery of its pathogenesis even more complicated. According to the World Health Organization (WHO), several environmental factors cause COLD, such as air pollution, occupational dust, and chemicals, and frequent lower respiratory infections during childhood. However, tobacco smoke is the primary cause of COLD. Therefore, all the recent COLD studies assessing the effect of environmental factors on the metabolic profile of patients are focused on tobacco smoke. For example, 37 healthy smokers, 41 COLD-smokers, and 37 non-smokers participated in a study aimed at discovering serum metabolic markers associated with early-onset COLD . It was The only known genetic factor affecting COLD is ZZ-a1-antitrypsin deficiency which was associated with changes in the pyruvate metabolism mediators including acetoin, propionate, acetone, and propane-1,2 diol in the EBC from a group of ZZ-a1-antitrypsin-deficient COLD patients .In addition to the establishment of specific biomarkers necessary for prognosis and accurate diagnosis of COLD, the analysis of the effect of different treatments on metabolites of COLD patients is also of critical importance. COLD patients receiving standard therapy in combination with doxycycline treatment presented disturbed levels of formate, citrate, imidazole, L-arginine, lactate, and fatty acids compared with COLD patients receiving only the standard therapy measured in serum by NMR . MoreovePhenotyping of COLD has been challenging for both researchers and clinicians due to the heterogeneity of the disease. Thus, some studies have been focused on identifying different metabolic biomarkers between different COLD phenotypes and pathological profiles . For exaN-acetylcysteine, guanosine, and lipoprotein only in type M COLD. However, there is some inconsistency in the metabolic profiles between studies on phenotype E and M COLD. For example, guanosine appears perturbed in both phenotypes in one case [Another characteristic example of discriminating different COLD phenotypes based on metabolomics is two studies on COLD patients with phenotype E and phenotype M. COLD phenotypes E and M are two out of the three phenotypes that were identified according to the dominance of emphysema and the presence of bronchial wall thickening based on high-resolution computed tomography (HRCT). Phenotype E exhibits apparent emphysema without bronchial wall thickening, while phenotype M exhibits apparent emphysema with bronchial wall thickening. These studies showed disturbed levels of ADP, choline, glycine, threonine, proline, tyrosine, ornithine, L-alanine, L-valine, L-leucine, fructose, pyruvate, and isopropyl alcohol in the serum measured by NMR in both phenotypes ,32. L-thone case . MoreoveCOLD is a chronic disease with various symptoms of the respiratory tract but also with a divergent metabolic fingerprint. A great number of COLD patients suffer from cachexia or metabolic syndrome, which has been strongly correlated to hospitalization and morbidity. The metabolic pressure in chronic diseases has been supported by several researchers stressing the need to find tools that can detect nutritional deficiencies, and metabolism disruptions that are related to the disease ,34.Untargeted metabolomic profiling of COLD patients has indicated several potential metabolite biomarkers that participate in aminoacid metabolism, lipid metabolism, and energy metabolism, as well as oxidative stress. From the pathways mentioned, the most commonly affected among the studies was the metabolism of aminoacids . Levels However, as already mentioned, there is some variability and inconsistency between the results of different metabolomic studies on COLD that hampers the validation of biomarkers. This could be explained due to different measurement techniques and standards used in each study, different sample preparation, handling and processing conditions, different quality control samples and internal standards, and different number of replicates used in each study that are rarely mentioned , differeThe major problem of modern medical science, according to the medical doctor Seth J. Baum, lies on the lack of communication between clinicians and researchers leading to the poor interpretability of human studies . In the As has been discussed elsewhere, biomarker discovery and validation should follow a multistep approach using untargeted metabolomics for the initial screening followed by quantitative analysis through targeted metabolomics and biological interpretation of the results  Figure . SpecifiImprovements and major future steps must be taken in the field of metabolomics in general, regarding more reliable and precise metabolite identification, data analysis and interpretation of the biological relevance of the biomarkers identified. As already mentioned, metabolite identification in untargeted studies remains the main bottleneck in data analysis and interpretation . In contThe goal of precision medicine is to provide tailor made treatment for all, considering the inter-individual variation in treatment response. Pharmacometabolomics is an emerging and rapidly evolving field regarding the identification of metabolic biomarkers or networks that will facilitate the treatment selection and appropriate dose. It was first introduced by Clayton et al., who developed a predictive model based on the metabolic changes of rats after treatment compared to the pre-treatment profile . Today tThere is accumulating evidence which suggests that nutritional deficiency plays a key role in COLD patients which partially explains the metabolic imbalances that are observed in the metabolomic profile of these patients . AlthougThe outstanding advantage of metabolomics is that the clinical doctor will be able to provide a personalized diagnosis and treatment according to the patient’s metabolic profile and not the symptomatology . There iOverall, even if great progress has been made in the field of metabolomics for COLD diagnosis and treatment, and a big range of metabolites are suggested as potential biomarkers for COLD most of them are still not validated. Common metabolic patterns of COLD patients include changes in the aminoacid metabolism and the ratios of certain aminoacid groups such as the BCAA/AAA ratio. These metabolic biomarkers have been associated with clinical symptoms, such as weight loss and reduced % FEV1, contributing to the selection of the right specialized treatment for each clinical case. However, large and longitudinal studies with a thorough analysis of the demographic variables of patients and the metabolic profiles will facilitate the validation of these biomarkers. The majority of the metabolomic studies were research-based, performing untargeted analysis of COLD patients and ending up with a large amount of metabolomic data that was difficult for scientists and clinicians to use. Tools of precision medicine, including HER and computational analysis, will help the management and analysis of data, and pharmacometabolomics will bring precision medicine closer to clinical practice.


There is evidence of a strong correlation between inequality in health care access and disparities in chronic health conditions. Equal access to health care is an important indicator for overall population health, and the urban road network has a significant influence on the spatial distribution of urban service facilities. In this study, the network kernel density estimation was applied to detect the hot spots of health care service along the road network of Shenzhen, and we further explored the influences of population and road density on the aggregate intensity distributions at the community level, using spatial stratified heterogeneity analyses. Then, we measured the spatial clustering patterns of health care facilities in each of the ten districts of Shenzhen using the network K-function, and the interrelationships between health care facilities and hypertension patients. The results can be used to examine the reasonability of the existing health care system, which would be valuable for developing more effective prevention, control, and treatment of chronic health conditions. Further research should consider the influence of nonspatial factors on health care service access. China has experienced a dramatic demographic change and epidemiologic transition in the past three decades . AlthougIn modern cities, health care facilities are part of the public infrastructure, providing beneficial functions for urban residents, including pre-hospital emergency medical care, primary care services, and community health care services. Based on China’s hierarchical administrative structure, the health care system is a service network composed of clinics, community health centers (CHCs), and hospitals, including primary hospitals, secondary hospitals and tertiary hospitals . This muThere is evidence of a strong correlation between inequality in health care access and disparities in various health outcomes, including low birth weight, maternal mortality, circulatory disease, cardiovascular events, late-stage cancer diagnosis, and others. Hypertensive disease is regarded as the most common non-communicable disease (NCD) in China, affecting over 200 million patients , but hypHealth care accessibility refers to the ease or difficulty of reaching services from a given location ,13, whicIn a GIS environment, health care facilities, just like many other geographical phenomena such as car crashes on a road, residential locations, crimes and disease outbreak sites, can be abstracted as points for spatial analysis. Point pattern analysis (PPA) has been widely applied to investigate the global or local spatial distribution patterns of point events ,21. In tDistance-related inequality in health care service use is potentially a significant public health issue, and the interrelation between spatial accessibility to health care services and the disparities in chronic diseases is particularly interesting in China. Therefore, this paper aims to fulfill two research objectives in a highly urbanized area of China: Shenzhen. The first objective is to measure the spatial distribution characteristics of health care facilities using network-based analysis methods. The second objective is to explore the interrelationship between health care facilities and hypertension patients in the road network space. The next section of this paper describes the materials and methods. 2). Shenzhen has become a modern metropolis with well-developed secondary and tertiary industries, and is one of the fastest-growing cities in the world. By the end of 2014, after the reclamation of some coastal wetlands, Shenzhen had a total population of approximately 20 million and the municipality covered an area of 1991.64 km2, including urban and rural areas [Shenzhen is located in the south of Guangdong province, bordering Dongguan in the north, Huizhou in the north and northeast, Hong Kong across the Shenzhen River in the south, Lingdingyang and the Pearl River in the west, and Mirs Bay in the east . The stual areas . The total areas . Shenzheal areas . Data on hypertension inpatients were provided by Shenzhen Center for Health Information, with 10,395 cases recorded in 2013. We obtained data for primary hypertension , hypertension heart disease (ICD-10 I11), hypertension renal disease (ICD-10 I12), hypertension heart and renal disease (ICD-10 I13), and secondary hypertension (ICD-10 I15). The hypertension patients were mapped to point events using geocoding tools  based onData on the health care facilities were obtained from the Urban Planning, Land and Resources Commission of Shenzhen Municipality, including 344 hospitals, 751 CHCs, 2035 clinics, and 5519 pharmacy stores that were abstracted as point locations for spatial analysis. We also included the bed numbers of the hospitals to represent their capacity to accommodate patients. Network kernel density estimation (NetKDE) is a nonparametric method that aims to examine the first-order properties of spatial data , which eandwidth ,38. ThusThe network K-function method uses the shortest-path distance to test the complete spatial randomness (CSR) hypothesis that points are independently and identically distributed according to the uniform distribution over the network, or points follow the homogeneous binomial point process on the bounded network . We applFor a set of points, t events ,33,40. TThe geographical detector method  is a spaThe value of the q-statistic is between We considered the influence of non-spatial factors of health care facilities in the analysis. Here, the intensity of hospitals was weighted by the bed numbers of each health care facility, representing the importance of the point event. According to the service capacity standards of CHCs and Shenzhen health statistics, CHCs, clinics, and pharmacy stores were assigned with weights of 20, 10, and 1, respectively. The computation of NetKDE was implemented in the ESRI ArcGIS 10.5 environment, using Microsoft Visual C# 2016. The choice of search bandwidth is an essential issue in KDE; a larger bandwidth may lead to unexpected generalizations, and a smaller bandwidth could over-emphasize local variation. Previous studies suggested that a 300 m bandwidth was suitable for the study of activities of urban residents ,44, whicIn the visualization of density surface, we applied Jenks classification based on the statistical characteristics of the NetKDE values. We further extracted the isolines of different classification numbers and then calculated those weight means of the isolines. After several rounds of experiments with different numbers of classification, we set the number of classes to five because there is no significant variation in the weight means of different numbers of classification.The sum of the NetKDE values of the health care facilities was aggregated at the community level. Spatial stratified heterogeneity analyses were further applied to explore whether population density and road density simultaneously affect the aggregate intensity of health care facilities, as estimated by NetKDE methods. The average population density and road density per square kilometer was 29,132.10 and 7350.59 m. The Spatial Analysis along Networks (SANET) toolbox  was applThe results of the network-auto K-function indicate that hospitals tend to be in a clustered distribution in Baoan a, DapengThe spatial distribution of the CHCs exhibits a different pattern. For the analysis of the spatial cluster pattern of the CHCs, The network cross K-function was applied to analyze the relationship between hypertension patients and health care facilities across 10 districts of Shenzhen. The results were used to investigate whether the configuration of one type of point could influence the distribution of another kind of point event. We first examined the interrelationship between hospitals and hypertension patients in the road network space across the Shenzhen Municipality. We further examined the effects of CHCs, clinics, and pharmacy stores on the spatial distribution characteristics of hypertension patients in the road network space, as shown in This study analyzed the spatial distribution characteristics of health care facilities in the road network space and their relationship with hypertension patients of Shenzhen using network-based spatial analysis. NetKDE was applied to detect health care service hot spots, and weighting of different types of health care facilities could be useful to obtain a comprehensive understanding of the intensity distribution of health care facilities. We further explored the influences of population density and road density on the aggregate intensity of different types of health care facilities at the community level, by using spatial stratified heterogeneity analyses. Then, the network K-function method was applied to measure the spatial clustering patterns in each of the 10 districts of Shenzhen along the road network. The spatial distribution of health care facilities displays distinct patterns in the 10 districts of Shenzhen, as well as in the interrelationships between health care providers and hypertension patients. Although the number of hospitals is relatively small, there is a stronger clustering pattern relationship between hospitals and hypertension inpatients along the road network. Due to the limitations of the data, directions of hypertension inpatient’s travel are not considered in this study and road segments are treated equally without being further distinguished. However, road segments are different in their traffic flow capacity, directionality, and so on, which may have an impact on the directions of travel of hypertension inpatients, subsequently affecting the density estimation of health care facilities and their interrelationships with hypertension inpatients. When the data becomes available, it may be beneficial to incorporate them into the analyses.In China, public hospitals dominate health care delivery, and patients can visit hospitals directly for all health care needs. This may lead to increasingly long lines and waiting times, which in turn, increase costs and decrease access to health care even though the hospitals are located close to residential areas. Strengthening primary health care could be a cost-effective way to detect, diagnose, treat, and manage hypertension patients, to alleviate the burden on higher-level health services and mitigate medical costs to both individuals and the health systems. However, given the difficulty in collecting data at an individual level, we cannot confidently assert that strengthened primary health care services would affect the health-seeking behavior of hypertension individuals. Nevertheless, our findings confirm the importance of primary health care facilities, which require long-term commitment from the department of urban planning and public health ,45. In this study, we investigated the interrelationship between hypertension patients and health care facilities in the road network space. However, the use of health care services is also influenced by nonspatial factors that include many demographic and socioeconomic variables , and which also interact with spatial accessibility . It is n