The purpose of this study was to investigate the association between smoking and health-related quality of life among Chinese individuals aged 40 years and older.Using a stratified multistage sampling method, data from 1,543 adults aged 40 years and older were obtained from a household survey conducted in eight provinces in China. The health-related quality of life was quantified based on the utility index obtained using a standardized instrument entitled “The European Five-Dimensional Health Scale (EQ-5D-5L).” Descriptive statistics were used to summarize the demographic characteristics and social factors of the sample according to smoking status. An instrumental variable (IV) probit model was used to estimate the association between smoking status and health-related quality of life.p < 0.01). For smokers, the average probability of having a higher quality of life was 11.65% lower than when they did not smoke.Of the 1,543 participants, 485 (31.43%) were smokers and 1,058 (68.57%) were non-smokers. Smoking was negatively associated with the probability of having a higher quality of life (These findings suggest that smoking reduces health-related quality of life among Chinese individuals aged 40 years and older. Anti-smoking programs should consider this factor. The tobacco epidemic is the greatest but preventable risk factor for human health. Approximately 8 million people worldwide die from smoking each year, and more than 80% of the 1.3 billion tobacco users worldwide live in low- and middle-income countries . China'sIn addition to health threats, smoking can also directly affect health-related quality of life (HRQoL) –7. As a It seems to be a common belief that smoking can help relieve stress and promote relaxation, thus creating the illusion that smokers have a much higher quality of life than non-smokers. However, accumulating evidence suggests that HRQoL is better among non-smokers and former smokers than among current smokers –10. The The association between smoking and HRQoL may have different manifestations in different countries, where the cultural context may be at play. Most related studies have been conducted in western countries, including European countries –21, the The challenges in studying the association between smoking status and HRQoL are sample self-selection and sampling bias. However, the possible endogeneity of the relationship between smoking status and quality of life has rarely been considered. Whether to smoke is a self-selective behavior that can be influenced by many factors. The omitted variables that may affect both smoking and HRQoL will make the results less credible.Therefore, the objective of this study was to explore the association between smoking and HRQoL among Chinese individuals aged 40 years and older using an instrumental variable (IV) probit model.N = 1,543) were Chinese individuals aged 40 years and older recruited in a household survey conducted in China between November 1, 2019, and October 30, 2020. A stratified multistage sampling method was used to select participants from 24 primary health care facilities. These 24 primary health care facilities were selected as follows: firstly, 8 provinces were selected in the east, middle, and west of China: Hebei, Heilongjiang, Shandong, Henan, Hubei, Sichuan, Guizhou, and Shaanxi. Then 2–4 primary health care institutions, including township health centers and community health service centers, were randomly selected in each province. From the areas of 24 primary health care facilities, ~100 households were randomly selected. To be eligible, participants from the 100 households had to live in local communities for at least 6 months, have a minimum age of 40 years, and be willing to participate in this study.Participants  general household information, including household type, total household income, and expenditure; (2) basic personal information of household members, including gender, age, and education level; (3) smoking, smoking-related knowledge and chronic diseases of household members; and (4) self-care ability and quality of life of household members.In this study, the investigators were designated by each investigation unit, and then the investigators were uniformly trained by the research team. The means of household inquiries were adopted by the investigators. Besides, each survey unit identified a contact person who was responsible for survey organization, implementation, quality control, and unified reporting to the subject group. Finally, the research team organized and coded the questionnaires in a unified manner. This study was approved by the ethics committee of the Capital Institute of Pediatrics, Beijing (ID: SHERLL2020017). And the study was conducted following the ethical principles of the Declaration of Helsinki.The dependent variable was “quality of life utility index.” The HRQoL was quantified based on the utility index obtained using a standardized instrument entitled “The European Five-Dimensional Health Scale (EQ-5D-5L).” The EQ-5D is easy to operate and easy to understand by the survey subject and has good reliability and validity. Therefore, it has been widely used in various research fields in many countries  and has The measure of smoking status came from responses to the following question: “Do you smoke now?” In the current study, the concept of smoking was defined as “having smoked at least 1 day in the past 30 days.” Response options were (1) No, (2) Yes, and (3) Have quit smoking. To facilitate statistical analysis, smoking status was dummy coded: (1) smoking: participants who smoke now; (2) non-smoking: participants who had never smoked or had quit smoking.Based on prior knowledge , covariaWhether the increase in cigarette prices reduced the number of cigarettes smoked (PRS) was used as an instrumental variable.The variables used in the regression analysis are listed and defined in Descriptive statistics were used to summarize the demographic characteristics and social factors of the sample by smoking status. Characteristics of smokers were compared with those of non-smokers using chi-square tests for categorical variables and Kruskal–Wallis one-way ANOVA tests for continuous variables. Descriptive data are presented as the mean for continuous variables and as percentages for categorical variables.To investigate the association between smoking status and HRQoL, an instrumental variable (IV) probit model was used to control for potential endogeneity problems. Whether the increase in cigarette prices had an effect on the number of cigarettes smoked (PRS) was used as an instrumental variable. It was chosen because it is expected to be correlated with smoking behavior but not directly affect quality of life, thus satisfying the instrumental variable exogeneity requirement. Covariates included age, gender, educational level, marital status, logarithm of annual household income, occupation, family size, health status, and province. The province variable was entered into the model as a dummy variable to give the province a specific intercept to capture sample clustering.The parameter estimates from the IV probit were further used by marginal analysis to estimate the average treatment effect on the treated (ATET) of smoking on HRQoL. ATET is the estimated average difference of the treatment and control potential outcomes in the treated population. ATET is useful when there is interest in the quantification of the treatment effect in observational studies in which no definite parameter can be used. Therefore, ATET was calculated to obtain more intuitive and practical results.The instrumental variable (IV) probit model is constructed below as:Indexi refers to the quality of life utility index of the respondents and Smokei, the independent variable of interest, is binary. β1 is the coefficient of interest, which provides the estimated effect of smoking on HRQoL. Z is a vector of demographic and socioeconomic variables. I represents the instrumental variable. γ, π1 and π2 are the vectors of parameters for the control variables that need to be estimated. μi and αi are normally distributed error terms in the equation and i denotes an individual respondent.Here, p < 0.05 were considered statistically significant.All statistical analyses were performed using Stata . Values of p = 0.005). The proportion of smokers among males was 54.41%, whereas the proportion among females was 7.76% (p < 0.001). The mean value of the logarithm of family income for smokers was 11.03 compared to 11.14 for non-smokers (p = 0.006). There was no significant difference concerning educational status (p = 0.268), marital status (p = 0.358), occupations (p = 0.098), and family size (p = 0.394) between smokers and non-smokers, whereas there were significant differences in the distribution of provinces (p < 0.001). Among the participants with two or fewer chronic diseases, 30.91% were smokers and 69.09% were non-smokers (p = 0.009).The characteristics of subjects are summarized in The middle column in p < 0.01). As shown in The right column in In Our study indicated that smoking led to a lower probability of having a higher quality of life. For smokers, the average probability of having a higher quality of life was 11.65 percent lower than when they did not smoke.These compelling findings confirm previous findings reported from other countries such as the UK , 32, USAThe reasons for the observed negative association between smoking and HRQoL can be attributed to the following aspects. First, smoking increases the risk of non-communicable diseases, including cancers and cardiovascular and respiratory diseases . Adults Besides smoking status that was previously discussed, age and gender were found to be independent variables of a lower HRQoL. This finding suggests that smoking intervention programs might be targeted for specific populations, such as men in particular age groups who are current smokers, in order to better improve HRQoL.Popular belief has it that quitting will decrease HRQoL—because individuals believe it interferes with relationships or produces a loss of smoking related pleasure (such as reducing stress or promoting relaxation). However, the current study indicated that smoking did not improve HRQoL as one would expect. This result contributes to the knowledge of the association between smoking and HRQoL. Knowledge of this association is useful for two reasons: (1) to assist the economic evaluation of cessation programs by providing a more direct measure of health outcomes than the cessation itself; (2) to provide a good reason for individuals to quit smoking.The strengths of this study are as follows. First, it was based on a large and nationally representative sample of middle-aged and older Chinese individuals. We were able to examine the association between smoking and HRQoL and control many potential confounding factors. The large sample size enabled sufficient power for statistical inference. Second, we used the IV probit regression model to address selection bias. The estimated results of the IV probit regression model were higher than the estimated results of the probit regression model, indicating that the probit model might underestimate the effect of smoking on HRQoL because of selection bias.There are several limitations to the present study. First, we relied on self-report measures, which may be subject to recall bias and social desirability effects. Second, the study was cross-sectional in design, thus making it hard to obtain any conclusions regarding exact cause-and-effect relationships. Longitudinal data may be needed to further explore the causal relationship between smoking and HRQoL. Third, the generalizability of our results to other populations is limited because we focused on China, and other countries may be different due to ethnic differences.Findings from the current study suggest that for smokers, the average probability of having a higher quality of life was 11.65% lower than when they did not smoke. Emphasizing that smoking will lead to a lower quality of life may help guide smokers to consciously quit smoking. Therefore, it is necessary for anti-smoking campaigns to clearly point out the negative effect of tobacco use on HRQoL.15107100093@163.com.The datasets presented in this article are not readily available because data involves personal privacy issues. Requests to access the datasets should be directed to Xi Cheng, XC: conceptualization, data curation, writing - original draft, and writing - review & editing. CJ: conceptualization, funding acquisition, investigation, supervision, and writing - review & editing. All authors contributed to the article and approved the submitted version.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Zero and low alcohol products, particularly beer, are gaining consideration as a method to reduce consumption of ethanol. We do not know if this approach is likely to increase or decrease health inequalities. The aim of the study was to determine if the purchase and consumption of zero and low alcohol beers differs by demographic and socio-economic characteristics of consumers. Based on British household purchase data from 79,411 households and on British survey data of more than 104,635 adult (18+) respondents, we estimated the likelihood of buying and drinking zero (ABV = 0.0%) and low alcohol (ABV > 0.0% and ≤ 3.5%) beer by a range of socio-demographic characteristics. We found that buying and consuming zero alcohol beer is much more likely to occur in younger age groups, in more affluent households, and in those with higher social grades, with gaps in buying zero alcohol beer between households in higher and lower social grades widening between 2015 and 2020. Buying and drinking low alcohol beer had less consistent relationships with socio-demographic characteristics, but was strongly driven by households that normally buy and drink the most alcohol. Common to many health-related behaviours, it seems that it is the more affluent that lead the way in choosing zero or low alcohol products. Whilst the increased availability of zero and low alcohol products might be a useful tool to reduce overall ethanol consumption in the more socially advantageous part of society, it may be less beneficial for the rest of the population. Other evidence-based alcohol policy measures that lessen health inequalities, need to go hand-in-hand with those promoting the uptake of zero and low alcohol beer. Alcohol use is a leading risk factor for ill-health and premature death ,2. The WPreviously, we have shown that the introduction of new lower strength beers resulted in British households purchasing fewer grams of ethanol overall . From 20Although there is some limited grey literature on the topic , we do nWe undertook binary logistic cross-sectional regression analyses to estimate the likelihood of drinking or buying zero alcohol beer or low alcohol beer by socio-demographic characteristics of respondents using online survey data and household purchase data.We used two data sources: (1) household purchase data that capture numbers of adults in the household, age of the main shopper, social grade group of the household, household income, level of area-based residential deprivation, and region of Great Britain ; and (2) survey data that captures for each respondent, sex, age, social grade, level of area-based residential deprivation, and region of Great Britain. These are described in detail below.We used data from Kantar Worldpanel’s (KWP) household shopping panel for the years 2015 to 2020, the years for which we had data. The shopping panel comprises approximately 30,000 British households at any one time, recruited via stratified sampling, with targets set for region, household size, age of main shopper, and occupational group. Households record all off-trade purchases from all store types, including Internet shopping, brought back into the home using barcode scanners. To be included in KWP’s final datasets, households must meet quality control criteria (meeting thresholds for data recording and purchasing volume or spend (based on household size) every four weeks), with some 90–95% of households included . In our The data included the truncated postcode of each household . Alcohol purchases are recorded daily. For each individual purchase, the data includes the type and volume of the purchase using 19 drink categories, the brand, and the alcohol by volume (ABV). The volume purchased was combined with ABV to calculate grams of alcohol purchased.We grouped households into: (I) four groups of the age of the main shopper ; (II) four occupation-based social grade groups (AB (‘highest’), C1, C2, DE (‘lowest’))with AB including higher and intermediate managerial, administrative or professional workers, C1 supervisory or clerical and junior managerial, administrative or professional workers, C2 skilled manual workers, and DE semiskilled and unskilled manual workers, state pensioners, casual and lowest grade workers, and unemployed with state benefits only based on the National Readership Survey ; (III) fWe classified three beer groups: zero-alcohol beer with an ABV = 0.0%; low-alcohol beer with an ABV > 0.0% and ≤3.5%; and all other beer with an ABV > 3.5%.We used data from Kantar WorldPanel (KWP) Alcovision survey for the years 2015–2018, the years for which we had data. Alcovision is an ongoing cross-sectional online timeline follow-back (TLFB) diary survey of the previous week’s alcohol consumption, with an annual sample of approximately 30,000 individuals aged 18+ years in Great Britain. In the diary-based approach, participants provide detailed data on the consumption of alcohol during the previous seven days for each day. Each drink by named brand is recorded, with the volume consumed and whether the consumed drink was purchased on-trade  or off-trade . Participants complete the survey only once, without repeated surveys. Quota samples based on age, sex, social grade group based on occupation, and geographic region are drawn from Kantar’s managed access panel. Invitations to participate are sent out on set dates and timed such that completion dates of the survey occurs every month, and each day of the year is represented in the data. Weights based on age-sex groups, social class, and geographical region are constructed using UK census data to ensure representativeness of British adults, with 18–34-year-olds oversampled.p = 0.462).Drink diaries were completed by 104,635 respondents  during the four years from 2015 to 2018, with an average of 512 diaries per week, (SD = 173), a rate which remained stable over the four-year period . The number of drinks consumed by brand is recorded separately for on-and off-trade, with information given on serving sizes in millilitres (mL). Drinks are categorized into 19 categories. For beer products, the brand-specific ABVs from household purchase data were used . There wWe grouped respondents into (I) male or female; (II) four age groups ; (III) the same four occupation-based social grade groups as for the household purchase data (AB (‘highest’), C1, C2, DE (‘lowest’)), based on the National Readership Survey , ScotlanWe classified three beer groups: zero-alcohol beer with an ABV = 0.0%; low-alcohol beer with an ABV > 0.0% and ≤3.5%; and all other beer with an ABV > 3.5%.We undertook separate binary logistic regression analyses for the purchase data and the survey data, using both univariate  and multivariate  analyses. For the purchase data, the dependent variables were binary variables at the level of the household, had or had not ever purchased any of zero or low alcohol beer on any day of purchase of any alcohol product. For the survey data, the dependent variables were binary variables at the level of the respondent, had or had not consumed any of zero or low alcohol beer during the previous week. For the purchase data, we added date of first alcohol purchase and number of days between first and last purchase of any alcohol as covariates. For the survey data, we added date of completing the survey as a covariate.For all analyses, we report odds ratios with 95% confidence intervals. All analyses were performed with SPSSv26  .Based on the household purchase data, for any day that a household made an alcohol purchase, 0.92% (95% CI = 0.91 to 0.94) of households reported at least one purchase of zero alcohol beer, 2.17% (95% CI = 2.16 to 2.18) of low alcohol beer and 29.32% (95% CI = 29.26 to 29.38) of all other beer, Households that were more likely to purchase zero alcohol beer were heavier purchasers of alcohol overall, and of beer with an ABV > 3.5%, were of younger age groups (highest odds for 35–44 year olds) and were relatively affluent . There was no consistent pattern by region of Great Britain.Households that were more likely to purchase low alcohol beer were also heavier purchasers of alcohol overall, and of beer with an ABV > 3.5%, and were more likely to be middle-aged (45–64 years old) than from a younger or older age group. There was no consistent pattern by household income, social grade, level of residential deprivation, or region of Great Britain.For purchase of zero alcohol beer, we also examined differences based on the social grade over time . The difBased on the survey data, 0.34% (95% CI = 0.33 to 0.36) of respondents reported consuming at least one drink of zero alcohol beer during the previous week, 1.57% (95% CI = 1.54 to 1.60) of low alcohol beer and 34.3% (95% CI = 34.0 to 34.6) of all other beer. The proportion for all other beer declined for the first two and a half years and then plateaued, Respondents that were more likely to have consumed zero alcohol beer during the previous week were heavier drinkers of alcohol, consumers of beer with ABV > 3.5%, men, of younger age groups and from ‘higher’ social grades, and, converse to the purchase data, from more deprived residential areas. There was no consistent pattern by region of Great Britain.Respondents that were more likely to have consumed low alcohol beer during the previous week were heavier drinkers of alcohol, consumers of beer with ABV > 3.5%, men, of younger age groups, from ‘higher’ social grades, and from more deprived residential areas. There was no consistent pattern by region of Great Britain.Although there were increasing purchases of zero alcohol beer and increasing consumption of low alcohol beer over time, we found relatively few occasions of purchases and consumption of zero and low alcohol beer throughout the time periods surveyed. For every purchase of zero alcohol beer, there were 32 purchases of beer with an ABV of 3.5% or more, and for every purchase of low alcohol beer, there were nearly 14 purchases of beer with an ABV of 3.5% or more. The differences in consumption during the previous week were even higher. For every consumption of at least one zero alcohol beer during the previous week, there were 101 consumptions of at least one beer with an ABV of 3.5% or more, and for every consumption of at least one low alcohol beer, there were nearly 22 consumptions of at least one beer with an ABV of 3.5% or more.Zero alcohol beer was more likely to be bought and drunk by those who generally bought and drank the most alcohol, those who bought and drank beer with an ABV > 3.5%, men, those with younger ages, and those with higher incomes and higher social grades. There were differences between purchase and consumption by area of residential deprivation. Respondents from less deprived areas  were more likely to purchase zero alcohol beer, but less likely to report recent consumption of zero alcohol beer . With univariate analyses, those who lived in the north of England were more likely to buy zero alcohol beer, with this difference no longer present in the multivariate analysis. Based on household level purchase data, results suggest the difference between higher and lower social grades increased over time between 2015 and 2020, favouring higher social grades.Low alcohol beer was more likely to be bought and drunk by those who generally bought and drunk the most alcohol, those who bought and drunk higher strength beer (ABV > 3.5%), men, those with younger ages, those with higher social grades, and those who lived in most deprived residential areas, and with no clear pattern by region of Great Britain.Thus, although the data were not consistent across all socio-economic measures studied, it seemed that the purchase and consumption of zero and lower alcohol beers was greater amongst younger and more, rather than less, affluent households and respondents, and that the difference favouring the more affluent increased over time. We also found that buying and drinking zero and low alcohol beer was more common amongst those who bought and drunk the most alcohol overall and the most beer, as opposed to those who bought and drank lower amounts of alcohol overall and of beer. Given that we have previously demonstrated that the purchase of zero and lower alcohol beers amongst such heavier buyers of alcohol is associated with buying less grams of alcohol overall, and thus likely health benefit , the datBeyond the qualitative studies mentioned in the introduction ,15, we aOur analyses have several important strengths and limitations. We obtained purchase data from a large number of households and survey data from a large number of respondents. Although relying on compliance at the household level, purchase data based on product bar codes, and verified via digital receipts, is, of itself, objective. The same cannot be said for the survey data which are based on subjective reports of drinking, with such subjective reports tending to underestimate consumption as measured by sales or other recorded data . The timAlthough quality control and compliance for the purchase data are regularly monitored by Kantar, with households only included in the final data set if they adhere to pre-assigned quality control criteria (meeting thresholds for data recording and purchasing volume or spend (based on household size) every four weeks), the data have limitations. Heavy drinkers, particularly male drinkers or those with no fixed address or living in communal establishments, are likely to be under-represented in household panel data ,32 and tAs with all survey-based research on alcohol, data cannot claim representativeness . RepreseThe data sets did not include underage drinkers. Finally, the data is just from one jurisdiction (Great Britain) and the findings may not, necessarily, be representative of other jurisdictions.Although there is a growing trend , the preA key finding from our study is that younger and more socio-economically advantaged households and respondents were more likely to purchase and consume zero alcohol beer in particular. As such, policymakers should be careful in simply prioritising the increased availability of zero and low alcoholic products, as they may be disproportionally beneficial for already better-off populations and could potentially increase health inequalities. Rather, promoting the purchase and consumption of zero and low alcohol products, that may lead to less alcohol consumption , needs t


Not all the coffee produced goes to the roasting stage, because non-compliant green coffee beans are usually discarded by roasters and the silverskin of the coffee is usually removed and discarded. In the present work, non-compliant green coffee beans and coffee silverskins were fully characterized from a chemical point of view. In addition, enzyme-assisted extraction was applied to recover a fraction rich in proteins and polyphenols, tested for antimicrobial, antityrosinase, and antioxidant activities. Non-compliant green coffee beans showed higher amounts of polyphenols, flavanols, flavonoids, and caffeine than coffee silverskins (which were richer in tannins). The enzymatic extraction of non-compliant coffee green beans produced extracts with a good protein content and with a consistent quantity of polyphenols. The extract showed antioxidant, antityrosinase, and antimicrobial activity, thus representing a promising strategy to recover defective green coffee beans. The antioxidant and antimicrobial activity of coffee silver skins is lower than that of non-compliant coffee green beans extracts, while the antityrosinase activity is comparable. Coffea canephora  and Coffea arabica (“Arabica”). Most of the world coffee production is made up of Arabica (69–74%), with the remaining 25–30% made up of Robusta [Global coffee production reached 170.94 million 60-kg bags in 2019, primarily produced in South America, and in Brazil (61.7 million 60-kg bags), followed by Vietnam and Colombia. The leading importer is the United States ($5.7 billion), followed by Germany and France ($3.3 and $2.8 billion) [Roasters select coffee green beans (CGBs) to eliminate those that do not meet the required quality standards (non-compliant). The quantity of CGBs discarded depends on the selection criteria of the suppliers and the quality standards applied. For example, these defective beans make up about 20% of total coffee production in Brazil and are separated from non-defective beans before being marketed . Then, tGreen seeds of Arabica coffee contain 46–53% insoluble polysaccharides , 8.5–12% proteins, 15–18% lipids, and 3–4.5% minerals .+, Mg2+, and Ca2+), with a low fat content [CSS contains 50–60% dietary fiber, 16–19% protein, 1.6–3.3% fat, and about 7% ash . The CSS content . Additio content . Consequ content . CSS ext content ) have al content . The pro content . The pac content .Overall, the recovery of coffee by-products has focused, up to now, mainly on the polyphenol and fibrous components. At the same time, protein content and further valorization have been much less studied. As mentioned above, peptides derived from the protein fraction could have interesting biofunctional properties. These peptides could also be synergized with polyphenols to obtain extracts with improved properties.A mild environmentally friendly approach for protein recovery, still little studied on coffee by-products, could use proteases to obtain a simultaneous extraction and hydrolysis of proteins from coffee by-products, possibly with simultaneous extraction of the polyphenols present. This approach would be a rather innovative application on coffee byproducts, as up to now, only enzymes that act on the carbohydrate fraction have been used to hydrolyze spent coffee ground . Only onTherefore, the aim of the present work was to apply for the first time a protease-assisted extraction to recover the protein fraction of the two most important by-products of the coffee industry, the defective CGB and the CSS. With this approach, peptide/polyphenol mixtures were obtained, which were characterized by their composition. As the target applications of these extracts are their possible use as antioxidant compounds in active food packaging films, as anti-aging compounds to be included in cosmetic formulations, and as a feed additive with antimicrobial properties, their antioxidant, anti-tyrosinase, and antimicrobial properties were also tested. The possible correlation between the observed biofunctional properties and the composition of the extracts will be discussed, with the general aim of exploring enzyme-assisted extraction as a possible way of valorizing coffee by-products.The general objective of the work was to characterize two coffee by-products (CGB and CSS), to evaluate the enzyme-assisted extraction technique for the simultaneous recovery of proteins and polyphenols and to evaluate some bioactive properties of the selected extracts. The experimental scheme of the present work is shown in Coffea arabica L.) beans, n-CGBs, from industrial selection processing , were obtained by subjecting coffee lots from Central and South America, Asia, and Eastern Africa to electronic sorting by means of color mapping as described in [C. arabica L.) silverskin, CSS, from the industrial roasting process, was supplied by illycaffè S.p.A.  in flakes of sizes from 2 to 5 mm with a bulk density of 0.18 g/cm3 and a particle density of 0.71 g/cm3. n-CGBs were sampled in September 2018, while CSSs were collected in October 2018.The raw materials were selected among the most interesting from the point of view of industrial waste management, and for the possible contents of proteins and bioactive molecules. Non-compliant green coffee (ribed in . Coffee n = 3) to determine the macronutrient composition of the selected food by-products. Proximal analysis was performed as described in [A proximal analysis was performed in triplicate (ribed in . Brieflyv/v) methanol by overnight shaking at room temperature and subsequently centrifuged at 5000 rpm for 10 min at room temperature. Obtained methanolic samples were used to assess total polyphenol [The phenolic compound and caffeine contents were determined in the raw materials and extracts obtained to understand their composition and to investigate potential bioactivities. The by-products, CGB and CSS, were dried at 80 °C in an oven up to 48 h and ground with an Ultra-Turrax blender . Aliquots of dried samples (0.40 g dry wight (DW) CSS and 0.46 g DW CGB, corresponding to 0.5 g (fresh weight) FW) were treated with in 5 mL of 95%  ± SD.All assay procedures were performed in duplicate in two technical replicates each. The results are expressed as the mean  under supercritical conditions. This supercritical fluid extraction (SFE) process has the advantage over traditional solvents that the extract can be used in food or medical and pharmaceutical products subject to high product quality standards. The substance dissolved in the fluid gas is separated isobarically and isothermally. This process is used when, for example, the food product needs to be freed from extractable substances, as applied for the degreasing of green coffee beans. For this purpose, the extraction sleeve of the extraction unit was filled with 3 kg  of ground coffee beans and extracted at a pressure of 300 bar and 50 °C. The fat content of the ground green coffee beans was then removed. The fat content of the ground green coffee beans was determined analytically in advance to calculate the expected extract quantity. The degreasing process in the extraction plant was continued (approximately 7 h) until the calculated extract quantity was approximately achieved.Defatting. COB. licheniformis, trypsin, pepsin, papain, and a combination of protease from B. licheniformis and papain), finally obtaining peptides.Protease-assisted extraction was performed to recover the protein fraction (and co-extracted polyphenols) from coffee silverskin, raw, and defatted green coffee beans. Protease-assisted extraction was performed as described in . In shorB. licheniformis, papain, trypsin, and mix) or 10 mM hydrochloric acid (pepsin).Reaction medium: 10 mM phosphate buffer (for protease from w/w or v/w).Enzyme/substrate ratio: 1% (Hydraulic module: 1 to 5 (1 part of sample and 5 parts of reaction medium) for CGB, 1 to 10 for CSS.Hydrolysis time: 2 h.○B. licheniformis: pH 6.5–8.5, T 60 °C.Protease from ○Trypsin: pH 7–9, T 37 °C.○Pepsin: pH 2–4, T 37 °C.○Papain: pH 6–7, T 65 °C.○B. licheniformis and papain: pH 6.5–7, T 62.5 °C.Mix of protease from pH and temperature for each enzyme:The reaction was carried out under the following conditions:As a control, the extraction was carried out using the same time, pH, and temperature conditions but without the enzyme.Polyacrylamide gel electrophoresis in denaturing conditions was performed to study the protein profile (and protein integrity) in the raw materials and the obtained extracts. The solutions obtained from the aqueous, and enzyme-assisted extraction were analyzed directly. Electrophoresis was performed as described in a previous work  and accoTotal amino acid analysis was performed to determine the nutritional value of proteins in the raw materials and extracts obtained, in terms of essential amino acids . The free amino acid content was determined in the extracts obtained to study the intensity of proteolysis during the extraction process. Total amino acids were determined after acid hydrolysis, as previously described , with slFor the analysis of free amino acids, 10 µL of sample were derivatized with the AccQ Tag Fluor reagent kit and analyzed with RP-UPLC/ESI-MS as previously described .The degree of racemization was determined to evaluate the quality of the proteins in the raw materials and extracts obtained. A high degree of racemization decreases the nutritional value of the proteins, and is an indication of microbial activity or severe processing conditions. The degree of racemization was determined by GC-MS after acid hydrolysis and derivatization, as previously described . BrieflyCoffea as the database.Protein identification was performed to identify the main proteins in the raw materials. The electrophoretic bands were gel digested with trypsin after reduction with dithio-threitol and alkylation with iodoacetamide and analyzed by liquid chromatography coupled with mass spectrometry , as described in a previous work . ProteinThe possible bioactivity of the extracts was determined by studying their antioxidant and antityrosinase activity. The extracts were resuspended in MilliQ water at a 200 mg/mL final concentration before biological activity determination.The extract’s in vitro antioxidant activity was measured using the 20-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) method with minor modifications . The resAntityrosinase activity was assessed by an optimized tyrosinase enzyme inhibition assay . The kint = 0 h. As pathogen, an exponential culture of E. coli F4 and Streptococcus suis  was used, at a dose of 106–107 CFU/g feed. The suspension was incubated under stirring for 3 h at 37 °C, to mimic the time effect in the stomach of the monogastric animal. Dilution series and bacterial counts were performed at the start (t = 0 h) of the experiment and after 3 h of incubation. Results are expressed as the relative kill-off related to a feed without extract.Antimicrobial activity was determined to evaluate the potential application for inclusion in feed for monogastrics. Antimicrobial extracts are highly interesting for this application, as they are potential alternatives for conventional antibiotics, which are still commonly used in today’s livestock production practices. The antimicrobial activity of the extracts was therefore determined in a monogastric stomach simulation model. A 20% feed suspension was made, analogous to monogastric livestock animals’ in vivo stomach conditions. To this suspension, the extract was added. The pH was controlled and set to pH 3.50 with HCl 1 N as is the case in vivo for the control and the trial group. A pathogenic strain was added to the blank and test suspensions at p < 0.05To highlight significant differences, statistical analysis was performed using IBM SPSS Statistic software (v. 26) . The ANOVA single factor test was applied to analyze the centesimal analysis data, with the significance threshold set at 0.05, with the aim of comparing the composition of the two raw materials (CGB and CSS). The Kruskal–Wallis test was applied to analyze data from the analysis of phenolic compounds and caffeine, with a significance threshold set at p < 0.05).The aim was to compare the content of these compounds between the two raw materials (CGB and CSS), and between the two extracts (degreased CGB and CSS) obtained with enzyme-assisted extraction. Regarding the protein extraction efficiency, the homogeneity of the variance of the data was verified with the Levene test and the Kruskal–Wallis test was applied to evaluate statistically significant differences between the samples  and coffee silver skins (CSSs). All samples belonged to the t-test, p < 0.05). However, it must be considered that the presence of caffeine (being a nitrogenous compound) can affect this result in both by-products. In addition, in CSS, the heat-induced reactions to proteins  may have transformed the proteins into pyrazines and other non-protein nitrogenous compounds. Both factors can lead to an overestimation of the actual protein content determined with the Kjeldahl method. All mean values of proximal analysis were not significantly different (p < 0.05) between CGB and CSS.The water content of the CSS was found to be higher than that of CGB. As a matter of fact, it was possible to store the CGB at room temperature, while the CSSs needed to be stabilized to avoid the formation of mold . The crude protein content was found to be comparable between the two by-products . To our knowledge, this is the first time that the degree of protein hydrolysis has been determined in these by-products. The DH% was very low for both raw materials (1.9 ± 0.1 for the CGB and between 0.4 and 0.9% for the CSS). This means that a negligible amount of free amino acids and peptides are formed by peptide bond hydrolysis. The low DH% for CGB confirms the protein integrity observed in gel electrophoresis. For CSS, the low DH% together with the absence of clearly defined protein bands on the gel may indicate the presence of protein-derived high molecular weight (MW)products (polymerization or aggregation phenomena), or extensive degradation, leading to a complex mixture of degraded nitrogen compounds.To further investigate the presence of possible protein hydrolysis, the degree of hydrolysis (DH%) of the samples was determined with OPA , confirming the high protein integrity also observed with SDS-PAGE and DH%. On the contrary, CSS showed higher values of d-Glu and, even more, d-Asp (close to 10%). The high temperatures used for roasting can explain this increased amount of d-Glu and d-Asp, as aggressive heat treatments can induce a certain degree of racemization. Consistently, very low racemization values were found for green coffee [As a further indicator of the protein quality of coffee by-products, the degree of racemization was also determined in all samples. Following a drastic heat treatment, or very acidic or alkaline pH conditions, a fraction of the amino acids, normally present in food in the n coffee . Going iThe real protein content  was found to be 8.8%  for CGB, and between 7.1 and 7.2% for the CSS. This is quite different from the values determined by Kjeldahl for both matrices see  above, iThe nutritional value of the proteins, calculated as an amino acid score, was found to be 0.5 for CGB (limiting amino acid Met) and between 0.4 and 0.5 for CSS (limiting amino acid Lys). Lys is sensitive amino acid, which easily degrades under roasting conditions, as it can be glycated during the Maillard reaction. The calculated amino acids scores are consistent with those of plant-based proteins.The by-product polyphenols and caffeine contents are reported in Overall, both matrices, and particularly CGB, showed a promising composition for consideration for further valorization.Initially, the CGB by-products underwent a deep pretreatment. First, CGB surfaces were scraped in order to improve the hygienic conditions of the sample. Then, to ensure even extraction, the beans were also ground. To prevent fat separation, the temperature during grinding was kept low, and thus the sample could not be ground to the desired fineness. Thus, to eventually improve further processability, CGBs also underwent alternative processing, including a defatting step of the grains. The details of both pretreatments are reported in the experimental section. In this way, pretreated non-defatted CGB and pretreated defatted CGB were obtained. As far as CSSs were concerned, no extensive pre-treatment, apart from grinding, was applied.Bacillus licheniformis protease, papain, pepsin, trypsin, and a mixture of Bacillus licheniformis protease and papain. The same extractions were performed precisely in the same conditions used for every enzyme extraction  but without the addition of the enzymes, in order to evaluate the efficiency of simple aqueous extraction as compared to the use of enzymes.Enzyme-assisted aqueous extraction (EAE) was applied to extract the protein fraction from these raw materials. EAE exploits the action of proteolytic enzymes that breaks down peptide bonds, releasing peptides and amino acids that have improved solubility compared to whole proteins. Several enzymes were tested: Extraction efficiencies were calculated as the amount of proteins extracted related to the initial amount of protein in the feedstock . While tBacillus licheniformis protease, which improved the extraction yield by 50%. Overall, both with and without enzymes, the protein extraction efficiencies appeared to be quite low, never exceeding 37%. The lower extraction yields obtained for CSS compared to CGB are consistent with the polymerization and degradation of proteins during roasting, leading to high-molecular-weight insoluble compounds.The above data show that fat removal was not helpful, as the protein extraction efficiencies were always lower for defatted CGB. Comparing the extraction efficiency in the presence and in the absence of the proteolytic enzymes, it is evident that the two processes gave very similar results in both raw and defatted CGB. Additionally, in a previous study, the increase in protein extractability using alkaline or neutral proteases was small . This miThe protein profile of the extracted fractions is shown in For CGB, the protein profile was found to be the same in raw c and defIn the case of aqueous extraction at acid pH (CPE), the two most intense bands disappeared. This could be due to the lower solubility of these proteins at the low pH (2) used for this extraction. Accordingly, the extraction performed at the same pH with pepsin (PE) also led to protein precipitation and consistently extracted less protein, producing very poor fractions with only low-molecular-weight polypeptides.Bacillus licheniformis protease + papain, discrete bands could be observed after extraction, indicating a non-complete hydrolysis, and thus a low enzyme efficiency. Only when using Bacillus licheniformis protease did the protein bands disappear, leaving low-molecular-weight polypeptides, and thus indicating that the proteolytic activity was effective. However, as already said above, this proteolytic efficiency did not translate to an increased protein extraction in the case of CGB.As per the other enzymes used, in the case of papain, the enzymatic extraction yielded protein profiles very similar to the extraction performed in the same conditions without the enzyme, confirming that in this case, the activity of the enzyme was very limited. Additionally, in the case of trypsin and the mix In the case of CSS b, all thDue to the comparable extraction yield and protein content of the AE extracts and respective EAEs, together with the similar protein profile on SDS-PAGE, the total amino acid profile of the obtained extracts was characterized only for the EAE extracts . Since tFirst, the amino acid content was used to calculate the actual amount of protein in the extracts , Ile (20–31% lower in extracts), Lys (25–46% higher in extracts), and Cys (19–53% higher in extracts). On the other hand, CSS extract was different to the raw material, with a lower amount of Pro, Val, Leu, and Phe, and higher amounts of Lys and Tyr.Bacillus licheniformis protease provided a higher AAS .The amino acid score of the EAE extracts was calculated using human milk protein as a reference and was approximately 0.5–0.6 for the CGB extracts (limiting amino acid Ile) and approximately 0.6–0.7 for the CSS extracts. While for CGB, the amino acid score was comparable for the different enzymes, for CSS, the The free amino acids content was also measured to further investigate the proteolytic action of the enzymes used for EAE , while very little information is available on amino acids, peptides, and proteins.Bacillus licheniformis protease from extracted defatted CGB. The choice of this sample containing this specific enzyme as an extracting adjuvant was due to the consideration that usually, bioactive properties are due to low-molecular-weight peptides and not to whole proteins. From the gel electrophoresis  [w/w, gallic acid) [B. licheniformis protease, which may have generated peptides with antityrosinase activity. Anyway, none of the identified peptides are yet known for any potential bioactivity  . Some diic acid) , the valic acid) . Even coic acid) . Howeveractivity . There iE. coli F4 and Streptococcus suis, was determined and expressed as the relative kill-off related to a feed without extract. The results of the antimicrobial activity test at two dosages  are presented in The antimicrobial activity of the extracts was also determined in a monogastric stomach simulation model. The antimicrobial activity against two pathogens, Streptococcus suis, the Gram-positive pathogen, at the highest inclusion level. Nanoemulsions containing oil and aqueous extract of green coffee beans have previously shown antibacterial activity against S. aureus, S. epidermidis, and E. coli [S. aureus) [S. aureus, P. aeruginosa, E. coli) [S. aureus, S. epidermidis, E. coli, and K. pneumoniae, thus indicating a potential antimicrobial effect [The defatted CGB extract showed promising antimicrobial activity against both the Gram-negative and Gram-positive pathogens. An increase in the inclusion level only gave rise to a marginal improvement in activity. This contrasts with the CSS extract, where a 10 times higher inclusion level caused an important increase in antimicrobial activity. The CSS extract showed promising antimicrobial activity against  E. coli , and sub aureus) . Thus, tE. coli) . Howeverl effect . TherefoB. licheniformis) produced extracts with a good protein content  and also with a consistent amount of polyphenols. The CGB extract showed antioxidant, antityrosinase, and antimicrobial activities, due to their polyphenol and peptide contents. This is a promising strategy to recover defective coffee green beans by producing extracts with interesting activities for the cosmetic or feed industry. CSS extracts are less promising than CGBs, as the quantity and quality of proteins are lower; the antioxidant and antimicrobial activity is lower than that in CGB extracts, while the antityrosinase activity is comparable to that of CGB. Therefore, a valorization path oriented more towards producing bio-based composites would be more suitable for CSS. More studies are needed to correlate the observed biological properties with the actual polyphenol and peptide composition.In the present work, a complete characterization of two by-products of the coffee industry, i.e., defective coffee green beans (CGBs) and coffee silver skins (CSSs), was carried out. The quantity and quality of the protein fraction, and the contents of polyphenols, flavanols, flavonoids, tannins, and caffeine were assessed. Except for the tannins, CGBs showed a greater amount of all these compounds than CSSs. Furthermore, the protein fraction of the CGB is very well conserved, while a strong degradation was observed for the CSS. Consistently, enzyme-assisted and conventional protein extraction from the two matrices yielded better results for CGB rather than CSS. The enzyme-assisted extraction of CGB (using the protease from


The present study was conducted to estimate the (co)variancecomponents for birth and weaning weight (BW and WW) in 8142 Awassi sheepbetween 2015 and 2017. Estimates were calculated with single-traitanalysis by the average information restricted maximum likelihood (AI-REML)method, using a derivative-free algorithm by fitting six differentunivariate animal models. The negative of the log-likelihood function(LogL), Akaike information criterion (AIC), and Bayesian informationcriterion (BIC) tests were used for selecting the best fitted model. Inaddition, the goodness of fit between the two models was compared with thelikelihood ratio test (LRT). Depending on the models,  Awassi sheep is one of the mostcommon breeds grown in Turkey . In addition, it is recognized asan important genetic resource for the sheep industry in more than 30countries in the Middle East and all over the world .Consumers in the Middle East prefer Awassi mutton due to its taste. Falconerand Mackay (1996) have reported that many nongenetic factors play animportant role in different quantitative characteristics. Birth weight (BW)and weaning weight (WW), which are quantitative characters, are importantselection criteria for prenatal and postnatal growth performance inlivestock breeding. In sheep breeding, the first 3-month weights duringweaning of sheep are important production characteristics . Many researchers reported significant effects oflambing year, sex, and maternal age on BW and WW .The yields to be improved by selection are affected by the geneticperformance of the individual and environmental factors. In addition toreducing the effects of environmental factors to increase the degree ofaccuracy in selection, accurate estimation of the components that make upthe genetic structure is required. In farm animals with long maternallifespans, such as sheep, the phenotype of the offspring is determined notonly by their genetic potential and environment, but also by the environmentprovided by their dams. The maternal effect on the offspring has been knownsince the beginning of animal breeding, and therefore it has been studiedincreasingly with developed software programs in recent years.Direct additive, maternal additive, and maternal permanent environmentalvariances are considered important factors that determine the lamb's geneticgain and therefore performance . Inthis connection, several studies have been done on heritability estimates and itsconstituent elements .The aims of this study were to estimate the variance and covariancecomponents, to compare different models for the BW–WW measurements of theAwassi lambs, and to reveal the importance of including maternal effects todesign true breeding programs for genotypic selection of Awassi lambs. Forthis purpose, direct additive, maternal genetic and the maternal permanentenvironmental effects were calculated by applying six animal models.22.1The study was carried out in Osmaniye province, districts, and villagesbetween Awassi is warm and very well adopted to arid conditions; it is one ofTurkey's indigenous breeds with a high ability to adapt to differentenvironments. Awassi sheep are raised in southeastern Anatolia, and the highmilk production is one of the remarkable traits of the breed. Newborn lambswere kept together with their dams for three months (90 d) until weaningage. The birth weight (BW) and weaning weight (WW) at 3 months of agewere recorded as growth traits. BW was taken within 24 h of the birth of lamb,and WW was taken on 3-month weights with digital hand weighing scales(up to 10 g sensitivity). The regional breeders kept their lambs insimilar feeding and management conditions. Except for the winter period,they apply a feeding with a nomadic system based on the pasture and stubblegrazing. They feed their animals mainly with hay and a small amount of grainin winter.The material of this study was obtained from the Awassi Sub-Project in whichelite herds were created in Osmaniye province within the scope of the nationalcommunity-based animal improvement project. The records collected from 249rams, 2416 sheep, and 8142 lambs between 2015–2017 years were used.2.2A preliminary least-squares analysis of variance was performed with a generallinear model (GLM) with SPSS software  to determine the maineffects that will take place in the individual animal model. The least-squares analysis showed that the effects of sex of lambs ,type of birth , year of birth , and age ofdam  were found to be statistically significant  method, using a derivative-freealgorithm by WOMBAT software . The matrix representation of sixsingle-trait animal models that used for genetic analysis were given asfollows:The total heritability . The most appropriate model forconvergence of REML solutions was assumed when the average difference inlikelihood functions in successive iterations was less than The log-likelihood (LogL) test, Akaike information criterion (AIC), andBayesian information criterion (BIC) were performed to determine the best model.These likelihood-based tests are 3The structure of the data set used in this study is presented in Table 1.According to descriptive statistics the coefficient of variation for WW ismuch lower than BW (Table 1).Least-squares means for fixed effects and standard errors for BW and WW areshown in Table 2, and they were estimated for BW and WW as Estimates of (co)variance components and genetic parameters for BWs ofAwassi lambs obtained with different models are presented in Table 3. Thedirect additive genetic variance was found to be lower in Models 2, 3, 4, 5,and 6 when compared with Model 1. Similar results have been reported forvarious goat and sheep breeds . Estimates of maternal permanent environmentalvariances in Models 4, 5, and 6 for BW were as 0.0145, 0.0073, and 0.001,respectively. The estimates of Correlations between direct additive genetic and maternal genetic effectswere calculated as In addition, the most suitable model was selected initalics as the best modelaccording to LogL, AIC, and BIC criteria for the estimation of geneticparameters for the BW and WW of Awassi lambs (Tables 3 and 4). Models 1,2, and 3 were mentioned as the best model according to BIC, AIC, and LogLcriteria, respectively. Also, Models 2, 3, and 4 were found to bemore statistically significant than Model 1 according to LRT (In Table 4 the results showed that Model 2 was selected as the bestmodel with respect to all criteria. Also, Models 2, 3, 4, 5, and 6 were found tobe significant compared to Model 1 according to LRT (In Model 4, 5, and 6 the The In present study, Tables 3 and 4 show that the estimates of maternal genetic and maternalpermanent environmental effects for WW were increased when compared with theestimates for BW. Also, Tables 3 and 4 show that the 4This research was conducted to compare different models based on predictionsof genetic parameters and to show whether maternal effects could betterexplain the genetic effects on BW and WW traits. Overestimation ofphenotypic variation, in other words, low total heritability, may be due tothe low litter sizes, inconsistency in records, and poor maintenance,feeding, and herd management in the farms. It is thought that by improvingthese factors, the reliability in estimations would be increased.Consequently, the maternal effect should be taken into consideration for thepurpose of improving accuracy in parameter estimations and thereforeincreasing the success of breeding programs.10.5194/aab-65-121-2022-supplementThe supplement related to this article is available online at: https://doi.org/10.5194/aab-65-121-2022-supplement.


June 2021 marks the 40th anniversary of the first description of AIDS. On the 30th anniversary, we defined priorities as improving use of existing interventions, clarifying optimal use of HIV testing and antiretroviral therapy for prevention and treatment, continuing research, and ensuring sustainability of the response. Despite scientific and programmatic progress, the end of AIDS is not in sight. Other major epidemics over the past decade have included Ebola, arbovirus infections, and coronavirus disease (COVID-19). A benchmark against which to compare other global interventions is the HIV/AIDS response in terms of funding, coordination, and solidarity. Lessons from Ebola and HIV/AIDS are pertinent to the COVID-19 response. The fifth decade of AIDS will have to position HIV/AIDS in the context of enhanced preparedness and capacity to respond to other potential pandemics and transnational health threats. However, the enthusiasm engendered by PreP science needs to be tempered by consideration of cost, need for rigorous adherence, rising rates of other sexually transmitted infections and thus need for continued condom use, and contraception for women. Long-acting injectables could be a major advance, but accessibility and logistics for their delivery need to be considered.Mathematical modeling and ecologic studies suggested that greatly increased delivery of ART could reduce HIV transmission at the community level. The definitive study showing that ART provided prevention benefits was the landmark HPTN 052 study  was an advocacy proposal rather than an evidence-based initiative, but these targets have become adopted as policy promising “epidemic control,” itself a concept requiring precise definition , WHO revised its International Health Regulations  to that of the broader Sustainable Development Goals   was completely unexpected; the literature on pandemic threats is voluminous. SARS in 2002–2003 was severe but not widespread; the 2009 influenza (H1N1) pandemic was widespread but not severe. It is hubristic to assume that pathogen severity and spread would always segregate, yet we were not prepared. Preparedness metrics can give false reassurance, witnessed by the lamentable response to COVID-19 in the United States in 2020. “Never again” was the mood after the Ebola epidemic in West Africa, but preparedness just seems too hard and costly. Perhaps true preparedness exists only in the military, where personnel train continuously for wars they hope will never happen.As a result of technologic advances such as whole-genome sequencing, scientific progress on COVID-19 has been breathtakingly rapid compared with early laboratory research on HIV. We hope to not see a replay of the early history of ART, with scientific advances relating to COVID-19, and specifically vaccines, not being rapidly or equitably accessible everywhere. “Vaccine nationalism” is a new term raising the specter of lower risk groups in high-income countries receiving vaccine before, for example, frontline healthcare workers in low-income settings. Healthcare workers have been disproportionately affected by Ebola and COVID-19, highlighting the need for much greater investment in infection prevention and control in healthcare settings worldwide. Attention and innovation are required to ensure maintenance of HIV and other essential public health services amid other outbreaks such as COVID-19.Although initially slow, the HIV/AIDS response over the years has been a beacon in global health for respect for individuals and their rights and for health equity. More reflection is required with regard to what the responses to HIV and Ebola have taught us and how they might be relevant to COVID-19 and other future epidemics.Although great need remains, the past decade has seen scientific and programmatic successes with regard to the HIV/AIDS priorities we defined after 30 years of AIDS. Existing interventions have been scaled up, and new tools such as PrEP and long-lasting drug preparations have been introduced. The roles of HIV testing and ART for treatment and prevention have been clarified, and the need for immediate ART for all HIV-infected persons has been proven. The global HIV/AIDS response has been sustained, financing has been maintained, and the world has kept focus on the SDGs. Mann’s judgment that “we did not separate ourselves” remains justified. We must also accept that political promises of “the end of AIDS” were hyperbole that current epidemiology does not support.The COVID-19 pandemic has exploited the fault lines of global systems and existing inequalities in a way that HIV did early on. Regrettably, the solidarity that HIV/AIDS engendered has not yet been carried over. In retrospect, the recent epidemics of Ebola in West Africa and DRC were preparation for the COVID-19 pandemic, but follow-through was lacking. The fifth decade of AIDS will take us to the SDG target date and reassessment of global health and development priorities. HIV/AIDS may not be central to global health discourse as it was earlier, but it will remain a yardstick by which to judge commitment and efforts, including, and especially in relation to, health security.https://www.who.int/emergencies/diseases/ebola/ebola-2021-north-kivu, https://www.who.int/csr/don/10-february-2021-ebola-drc/en).On February 7, 2021, the Ministry of Health of DRC reported a laboratory-confirmed case of Ebola in North Kivu Province, the most heavily affected province during the 2018–2020 outbreak in eastern Congo. The case-patient experienced symptom onset on January 25, 2021, and died in Butembo, a city of ≈1 million persons, on February 4, 2021. She was reportedly linked epidemiologically to an Ebola survivor, and genetic sequencing reportedly showed phylogenetic association with the earlier outbreak rather than a new spillover event. As of February 8, 2021, a total of 118 contacts were being investigated .Separately, on February 14, 2021, the Ministry of Health of the Republic of Guinea reported an outbreak of Ebola in the subprefecture of Gouécké, Nzérékoré Region, the first report of Ebola in Guinea since the 2014–2016 epidemic. The index case-patient, a nurse, experienced symptoms on January 18, 2021, and died on January 28, 2021. A total of 6 secondary Ebola cases were reported, 1 in a traditional practitioner who cared for the index case-patient and 5 in family members attending her subsequent funeral. Of the 7 case-patients, 5 died. As of February 15, 2021, a total of 192 contacts were being investigated, including in the capital city, Conakry (


Nephrotic syndrome (NS) results in massive proteinuria and hypoalbuminemia, which are responsible for a compensatory increase in protein synthesis in the liver. Serum cholinesterase (ChE) also increases in NS. However, its clinical value is not fully elucidated.In this study, 184 patients with NS who underwent kidney biopsy were included. The patients were divided into two groups according to serum ChE levels, as follows: hypercholinesterasemia (HC) and non-hypercholinesterasemia (NHC) groups. The clinical factors were compared between the two groups.The HC group had significantly more severe proteinuria and higher prevalence of high selective proteinuria than the NHC group. Furthermore, the prevalence of minimal change nephrotic syndrome (MCNS) was significantly higher in the HC group than that in the NHC group. Multivariate analysis revealed that the severity of proteinuria and MCNS were significantly associated with HC.In this study, HC in NS was associated with the severity of proteinuria and MCNS, and could help clinicians predict the histological diagnosis of NS.The online version contains supplementary material available at 10.1186/s12882-022-02764-0. Nephrotic syndrome (NS) is a glomerular disease characterized by severe proteinuria, hypoalbuminemia, and edema , 2. NS cCholinesterase (ChE) is one of the enzymes produced in the liver and belongs to the family of serine hydrolases . ClinicaTo resolve this issue, we investigated the clinical value of HC in NS.This study adopted a single-center retrospective observational study design, and adult patients with new-onset NS who underwent kidney biopsy at our hospital from April 2010 to March 2019 were included. We excluded patients under the age of 18 years, those with insufficient data on serum ChE, and those with decompensated liver cirrhosis. Finally, 184 subjects were included in this study. Our protocols were approved by the Kobe University Clinical Research Ethical Committee (no. B190327) and performed according to the recommendations of the Declaration of Helsinki for Biomedical Research involving human subjects. The Kobe University Clinical Research Ethical Committee approved to waive off the need for an informed consent and to use the opt-out approach in the study, because the data were retrospectively and anonymously analyzed.All clinical data were collected from the patients’ medical records. Urine and blood samples, including ChE, were measured using standard laboratory techniques . Serum ChE activity was determined using an assay with p-hydroxybenzoylcholine iodine salt as the substrate . HC was defined as elevated ChE levels above the upper limit of normal reference ranges in our hospital . All subjects were divided into two groups, as follows: the HC and non-HC (NHC) groups. Subsequently, we compared clinical characteristics between the two groups. Furthermore, in order to investigate the association of HC with proteinuria and histopathological diagnosis, all the patients were divided into six groups according to the tertiles of proteinuria levels  and the histopathological diagnosis (MCNS and non-MCNS), and we compared the prevalence of HC in each category.The histopathological diagnosis of kidney biopsy was determined in our pathology department. NS was defined as proteinuria ≥3.5 g/day or ≥ 3.5 g/g Cre and serum Alb ≤3.0 g/dL, according to the guidelines of the Japanese Society of Nephrology . As for P-values of less than 0.05 were used to denote statistical significance. All statistical analyses were performed using Statistical Package for the Social Sciences, version 25.0 .Data are expressed as mean ± standard deviation, median and interquartile range, or proportions. Differences between the two groups were tested using the unpaired t-test, Mann–Whitney U-test, and chi-square test, as appropriate. Furthermore, we conducted a logistic regression analysis for HC. Table p <  0.05) and higher prevalence of high selective proteinuria than the NHC group ; NHC: 4.3 g/day )  and MCNS ) (p <  0.05).Univariate logistic regression showed that HC was significantly associated with the following factors: age, mean blood pressure, estimated glomerular filtration rate, serum Alb, T-Chol, proteinuria severity, high selective proteinuria, and MCNS Table . After aTo clarify the association of HC with proteinuria and histopathological diagnosis, the patients were divided into six groups according to the tertiles of proteinuria levels and histopathological diagnosis (MCNS and non-MCNS), and we compared the prevalence of HC in each category. The prevalence of HC increased along with more severe proteinuria in both the MCNS and non-MCNS groups, and the prevalence was significantly higher in the MCNS group than that in the non-MCNS groups, across all tertiles of proteinuria Fig. . InteresIn this study, we demonstrated the following results: (1) the incidence of HC was approximately 40% in patients with NS, (2) the HC group had significantly more severe proteinuria and higher prevalence of high selective proteinuria than the NHC group, (3) the prevalence of MCNS was significantly higher in the HC group than that in the NHC group, (4) multivariate analysis revealed that HC was significantly associated with proteinuria severity and MCNS, and the prevalence of HC was significantly higher in the MCNS group than that in the non-MCNS group, irrespective of proteinuria severity.N = 16) than those in the healthy group [N = 19) had HC [NS can increase serum ChE levels, which is based on the results of previous studies , 14. Way) had HC . HoweverFirst, the HC group had significantly more severe proteinuria than the NHC group. The hepatic synthesis of various proteins, including Alb and ChE, is enhanced in response to the loss of Alb from the kidney among patients with NS , 8, 12. Second, our results demonstrated that the HC group had a significantly higher prevalence of high selective proteinuria than the NHC group. The results of a study have indicated that patients with highly selective proteinuria had a more intensive loss of Alb into the urine than those with less selective proteinuria, even if both have the same proteinuria severity . As statThird, histopathologically, the proportion of patients with MCNS was significantly larger in the HC group. We attempted to explain this result based on its clinical characteristics. The characteristics of MCNS include highly selective proteinuria , 17 and In daily clinical practice, the histopathological diagnosis of NS is critical information for determining the indication of treatment and subsequent prognosis. The standard method for the histopathological diagnosis is a kidney biopsy. Since it is an invasive procedure and cannot be routinely performed, an alternative diagnostic approach is also important. The histopathological diagnosis is predicted based on clinical information, such as the onset of NS and the selectivity of proteinuria; however, the SI of proteinuria may not always be easily obtained. In contrast, ChE measurement is simple and quick and can be performed in many institutions. This study demonstrated the relationship between HC and MCNS, and therefore, we considered that ChE facilitates the prediction of NS tissue types in clinical settings. T-chol also can be easily measured, and the absolute value of both T-chol and ChE was significantly higher in MCNS compared to non-MCNS in this study. However, Fig. This study has some limitations. (1) The sample size was small, and the logistic analysis had limitations. However, we believe that this study is valuable since only a few studies have investigated the clinical value of ChE in patients with NS. (2) There were no data on ChE before the onset of NS because of the retrospective observational nature of this study. (3) The number of patients with NS other than MCNS and membranous nephropathy was relatively small. To overcome these limitations, further examination is required.In conclusion, our results suggested that HC was associated with severe proteinuria and MCNS in NS and could help clinicians predict the histological diagnosis of NS.Additional file 1.


Acanthosis nigricans (AN) is a clinical sign that commonly occurs in obesity; however, its specificity and sensitivity have been controversial. It is unknown if AN severity degree can be a useful marker for cardiometabolic disorders screening. We suggest that the stratified analysis of AN severity degree in neck by Burke's scale could be a useful tool in the screening of cardiometabolic alterations in obese children.  The aim of this study was the association of AN severity degree in neck by Burke's scale with anthropometric, biochemical, and inflammatory parameters in obese school-age children from Mexico City. x2 test for frequencies and one-way ANOVA with Bonferroni posttest for continuous variables. Linear regression analysis adjusted by gender, BMI, and age was performed to evaluate the association between AN severity degree and cardiometabolic alterations. Statistical significance was set at p < 0.05.  A cross-sectional study was conducted, including 95 obese school-age children stratified by AN severity degree in neck by Burke's scale. Anthropometric and fasting biochemical measurements were determined. Variables were compared by p=0.001) and triglycerides (p=0.02) significantly increased and adiponectin significantly decreased (p=0.02). Positive associations between grade 3 AN and waist circumference, HOMA-IR, triglycerides, total cholesterol, and LDL cholesterol were observed.  As AN severity degree in neck by Burke's scale increased, diastolic blood pressure ( Our findings could be used to identify an easier clinical tool to prevent obesity progression and its complications in pediatrics. There are no similar studies. Obesity is an important public health issue in the world. In Mexico, the combined prevalence of overweight and obesity in the school-age population was 34.4% in 2012 and 32.2% in 2016, and high estimates represent an urgency in the care and prevention of complications from the early stages of life . SpecifiAcanthosis nigricans (AN) is recognized as the most frequent dermatosis in obesity. Clinically, AN lesions are described as thickened, symmetrical grayish-brown hyperpigmentation plaques. Histopathologically, papillomatosis and hyperkeratosis are observed. AN is often found in folds and areas of flexion and is most frequently located in the neck . AN is aDifferent epidemiological studies have reported AN incidence of approximately 7% in general population, but it can be present in up to 74% of obese people .Few AN prevalence studies in school-age children have been reported; therefore, AN should not be underestimated, especially in obese children, who are vulnerable to cardiometabolic disturbances . HoweverWhile several classification systems can be used to diagnose AN , 27. BurThis cross-sectional study was conducted between February and June 2016. Participants were collected using a convenience sampling method by nonprobabilistic consecutive selection, out of a total of 1,500 children registers from a database. We included 95 school-age children with obesity diagnosis from 4 recreational and social centers in Mexico City , Instituto Mexicano del Seguro Social (IMSS) insured, who reported anthropometric and biochemical parameters, blood pressure, HOMA-IR, adiponectin, leptin, and AN severity degree in neck by Burke's scale registered in the database. The sample was stratified into 5 groups according to AN severity degree in neck by Burke's scale on a 0–4 level .Patients with a history or current diagnosis of any type of cancer, thyroid disorders, psoriasis, atopic dermatitis, autoimmune diseases, endocrinological diseases, liver disease, users of supplements, antioxidants, steroids, antihistamines or diet, physical activity, or under pharmacological weight loss treatment were excluded. Children with BMI ≥98th percentile were also excluded.All participants were weighed using a digital scale , and height was measured using a portable stadiometer (Seca). Waist circumference (WC) of each participant was measured at the midpoint between the lowest rib and the iliac crest after a normal exhalation in the standing position. Obesity was determined using the calculated body mass index (BMI) and classified according to the standards reported by the Center for Disease Control  in 2000 (obesity ≥95th percentile of BMI) .Blood pressure (BP) was measured using a mercurial sphygmomanometer  with the appropriate cuff size for arm length following the North American Task Force 2004 Guidelines. The BP was measured in the right arm with the participant in a sitting position. Two BP measurements were obtained with a resting time of 5 min between each measurement. The mean of the two measurements was used in the analyses .Burke's scale was used to determine the severity of AN in the neck. When AN was not detectable on close inspection, the severity was considered grade 0. Grade 1 severity was defined as AN is clearly present on close visual inspection but not visible to the casual observer and not measurable. Grade 2 severity was defined as AN limited to the base of the skull that did not extend to the lateral margins of the neck . Grade 3 severity was defined as AN extending to the lateral margins of the neck and the posterior border of the sternocleidomastoid  that was not visible when the participant was observed from the front. Grade 4 severity was defined as AN extending anteriorly  that was visible when the participant was observed from the front .μU/mL))/405]  , 34.This study was approved by the Research and Bioethics Committee of the IMSS (registration number R-2012-785–071). Written informed consent was obtained from the parents of each participant prior to the study.Variables are reported as means and standard deviations, as well as percentages, and the normality distribution was evaluated by the Shapiro–Wilk test. Variables were compared using a chi-square test for frequencies and one-way ANOVA with Bonferroni posttest for continuous variables to assess the significance of differences in Burke's scale 0 versus 1, 2, 3, and 4. To evaluate the association of the severity of AN in the neck with WC, HOMA-IR, and the lipid profile, a linear regression analysis adjusted by sex, BMI, and age was performed.TM 3.3.2  software. Statistical significance was set at p < 0.05 [History of type 2 diabetes, overweight/obesity, age, and sex were analyzed first and variables that resulted in <10% change were not included in the subsequent analyses. Each covariate and its effect on the estimator were evaluated, and the final model included overweight or obesity, age, and sex. All analyses were conducted using Stata, SE v11.0  and Epi Infop < 0.05 .p=0.001) and triglycerides (p=0.02) were significantly increased in the groups as the severity of AN increased. Adiponectin concentration significantly decreased (p=0.02) as AN severity degree in neck by Burke's scale increased. BMI, WC, insulin, HOMA-IR, total cholesterol, LDL-C, and leptin increased as the severity of AN increased and the glucose and HDL-C decreased as the severity of AN increased, though the differences between the groups were not significant.A total of 95 children were analyzed. The participants' characteristics are listed in Linear regression analysis was performed to evaluate the association of AN severity degree in neck by Burke's scale with WC, HOMA-IR, and the lipid profile is listed in p=0.02), HOMA-IR , triglycerides , total cholesterol , and LDL-C .Our results showed that grade 3 of AN severity degree in neck by Burke's scale was associated with WC  obese, school-aged participants presented AN in the neck. Among the participants with a family history of type 2 diabetes, 66.66% had ≥ grade 1 AN in this study, which is higher than that previously reported in the San Antonio Family Diabetes Study , which fIt has been reported that up to 30% of obese patients are metabolically healthy, have insulin sensitivity like healthy individuals, normal weight, decreased visceral fat, and lower cardiometabolic risk factors ; howeverIn this study, positive associations between the presence of AN and variables associated with obesity were observed, which is consistent with previous studies that reported dyslipidemia and IR in school-age children in Mexico , 30. ComMartínez-Rojano et al. reported increased systolic and diastolic BP in pediatric patients with AN and obesity compared to patients ≤ the 90th percentile for weight, though AN severity degree was not analyzed. In our study, diastolic BP increased with an increase of AN severity degree in neck by Burke's scale . Though No statistically significant differences in fasting glucose concentrations were found between the groups in this study, which may be due to the higher fasting hyperinsulinism in participants with a higher increment of AN severity degree in neck by Burke's scale. The presence of hyperinsulinism in obese patients is usually associated with IR (measured as an increase in HOMA-IR) due to the increased insulin-like growth factor-1 (IGF-1) cell receptors in states of hyperinsulinemia. These receptors promote the epidermal growth factor receptor (EGFR), fibroblast growth factor receptor (FGFR), and the proliferation of epidermal keratinocytes and dermal fibroblasts when activated , 45. AlmThe presence of metabolic disorders is also represented by an inverse relationship between leptin and adiponectin concentrations in obese patients . It is uγ transcription factor, and NF-ƙβ in various tissues. These signals are transduced via the adiponectin receptor signaling pathway [Adiponectin has anti-inflammatory properties and downregulates the expression and release of several proinflammatory immune mediators. A leptin/adiponectin imbalance may be an important risk factor of developing type 2 diabetes and cardiovascular diseases associated with abdominal obesity . Adipone pathway , 61 and  pathway .To our knowledge, this is the first study to report that AN severity degree in neck by Burke's scale grade 3 is related to metabolic changes in obese children, suggesting that AN severity degree in neck by Burke's scale can be used as a clinical predictor for IR and hypertriglyceridemia that allows for timely treatments through changes in lifestyle, supplements, or medications .However, this study has some limitations as follows:(1) We recognize that our study group analyzed was small due to the type of sampling; however, there are no similar studies reported previously and it could be considered as a pilot, suggesting that it be carried out on a large scale in the future by increasing the size of the sample in order to determine sensitivity as a clinical tool of AN severity degree in neck by Burke's scale in obese children for screening for cardiometabolic alterations. Despite the size of the sample, our study showed statistically significant associations. (2) We did not perform an analysis of consumption frequency or 24 h dietary recall, which could infer the results; however, none of the included participants carried out any type of pharmacological or nonpharmacological treatment such as diet, physical activity , or lifeWe assume that AN severity degree in neck by Burke's scale could be used as a screening biomarker to obesity complications in vulnerable children; however, future research is required to identify its application through interventions, aimed at preventive measures for screening and early detection of obesity complications, including insulin resistance, dyslipidemia, and impaired fasting glucose, as well as the development of specialized multidisciplinary teams responsible for implementing the findings.