To adapt communications concerning vaccine prevention, we studied knowledge, beliefs and practices around meningitis risk and prevention in a young adult population in Burkina Faso in 2016, 5 years after the MenAfriVac® mass campaign and one year before the vaccine’s inclusion in the infant immunization schedule.In a representative sample of the population aged 15 to 33 years (N = 220) in Bobo-Dioulasso, Burkina Faso, study nurses administered a standardized paper questionnaire consisting of predominantly open questions, collecting information on meningitis risk factors and prevention, and on exposure to dry air and kitchen fire smoke. We identified themes and analyzed their frequency. We created a meningitis knowledge score (range 0 to 4) based on pre-defined best responses and analyzed the determinants of knowledge score levels ≥2 (basic score) and ≥3 (high score) using multivariate logistic regression.Biomedically supported facts and good practices were known by the majority of participants . Younger women aged 15–20 years had a higher frequency of low scores <2 (17.0%) compared to older women aged 21–33 years (6.3%) and men of both age groups (3.8%). Junior secondary School attendance explained the differences between the two groups of women, the gender gap for the older, but not the young women, and explained score differences among young women. Local understandings and practices for risk and prevention were commonly reported and used .This study shows a gender gap in knowledge of meningitis risk and prevention, largely due to education-level inequalities. Women below 21 years had particularly low levels of knowledge and may need interventions outside schools and perinatal care. Our study suggests a strong adherence to local understandings of and practices around meningitis risk and prevention, which should be taken into account by vaccination promotion. The African meningitis belt, which spans the sub Saharan African Sahel region from Senegal in the west to Ethiopia in the east, is characterized by a high incidence of bacterial meningitis. A long dry season combines with a meteorological phenomenon in which the Harmattan wind from the northeast brings dry air with high mineral aerosol load. As a consequence, African populations inhabiting the meningitis belt are exposed to conditions of low relative humidity and dust clouds of large and fine mineral particles . These cHaemophilus influenzae type b conjugate vaccine and 13-valent pneumococcal conjugate vaccine in the Expanded Programme on Immunization (EPI). In 2010, a serogroup A conjugate vaccine (MenAfriVac®) was introduced through mass campaigns among persons aged 1–29 years with the aim of interrupting meningococcal circulation in the population and providing individual immune protection for several years [Several vaccines against meningitis are used in the belt populations, including al years .The MenAfriVac® mass campaigns were accompanied by a large mobilization of civil society and communication efforts. The experience of recurrent meningitis epidemics with high attack rates (up to 1% of the population affected during an epidemic), high case fatality  and substantial economic burden due to treatment and frequent long-term sequelae  lead to Few scientific publications have described the populations’ beliefs with regard to meningitis and its vaccine prevention. A qualitative study in different towns in Burkina Faso before the MenAfriVac® campaign described the frequent parallel consultation of modern and traditional medicine and spiritual practice for meningitis treatment . A case-S1 Table). The number of responses were not limited and participants could give more than one response per question. We designed this questionnaire based on a pilot questionnaire administered during the second seroprevalence survey in 2013 and anthropological and public health expert knowledge. The questionnaire was developed and filled out in French. During training of the field workers and study nurses, a translation to the two most prevalent local languages (Dioula and Moré) was defined by the field team and used during the study interviews. Answers were written in a paper questionnaire and entered into a database by one data manager. Study nurses were consulted during entry in case of unclear interpretation of wording. Institutional Review Board approval was obtained from National Ethics and Health Research Committee of Burkina Faso, and the Institutional Ethics Committee of Centre Muraz, Bobo-Dioulasso, Burkina Faso.As described in detail elsewhere, we conducted a series of three seroprevalence surveys  in Bobo-Dioulasso, the second largest town of Burkina Faso . During In the present analysis, only participants aged 15 years old or older were included. Regarding beliefs and practices, we identified themes and assigned keywords, based on the free-text responses provided to open questions. During this process, all responses to a given question were listed and relevant themes identified . We created variables for the mention of each theme , which then allowed conducting statistics on the frequency of the theme.We conducted descriptive analyses on the frequency of different themes, stratifying by age (15–20 years vs. 21–33 years) and gender. Two-sided Fisher’s exact test was used to assess the significance of differences.S1 Text): persons at highest risk of meningitis: children (and “young people”); the effect of dry air on health: meningitis; mode of infection with meningitis: (droplets from) person to person; prevention: vaccination. We allocated one point for each correct response, thus the knowledge score ranged from 0 to 4. We analyzed the association of knowledge score levels ≥2 (basic score) and ≥3 (high score) with participant characteristics (education and proxies of socioeconomic status) using bivariate and multivariate logistic regression. Variables contributing at P<0.20 in bivariate analyses were included in a multivariate model and stepwise exclusion used based on their P-value, leaving only variables contributing at P<0.05. The overall contribution of variables with more than two modalities was assessed using the Wald test. Based on the hypothesis that gender and age (≥/< 21 years) interact in their effects on knowledge, we built an additional mode including a combined variable distinguishing younger women, younger men, older women and older men (M1). We then evaluated whether at least one year of junior secondary school explained the observed association between knowledge, age and gender (M2). In a third model (M3), we evaluated specifically among young women (the group with the lowest knowledge scores), whether junior secondary school education and satisfaction with own information level were associated with knowledge score. Odds ratios and exact P-values are shown. All analyses were carried out using STATA software version 14. Data is available as supporting material.With regards to meningitis pathophysiology, risk factors and prevention identified in epidemiological studies, we created a meningitis knowledge score using one pre-defined biomedical best response per question (Table 1). Recall-based and document-confirmed MenAfriVac® vaccination during the 2010 mass campaign  was found for 64.6% and 13.2% of participants, respectively. About half of participants (46.4%) were students, 72.4% declared being able to read a text in French language, and 64.6% had attended at last one year of junior secondary school (“collège”). Educational indicators showed a large gender disparity: 20.5% and 33.7%, respectively, of 15-to 20-year-old and 21-to 33-year-old women reported being illiterate, vs. 2.6% and 8.3%, respectively, among the corresponding male age groups . Attendance of at least one year of junior secondary school was reported for both female age categories as 66.0% and 46.9% respectively, vs. 84.6% and 86.8%, respectively, for men (Table 1). A gender gap was also observed for ≥1h of daily exposure to kitchen fire smoke, which was 40.4% and 33.3% among women, vs. 5.1% and 7.9% among men. By contrast, the percentage sharing dinner with >10 persons  and living in a household with an open air kitchen or one located in a hangar (52.3%) was similar among gender groups. The most commonly cited fuel for cooking was charcoal (40.9%), followed by firewood (24.1%) or combinations of firewood with other fuels (18.0%). Only 8.6% of households used gas exclusively. All households reported having a radio.Complete information was available for 200 of the 223 participants aged 15 years or older. Among them, age ranged from 15.0 to 33.4 years . There were 47 women  and 39 men (17.7%) aged 15–20 years, and 96 women (43.6%) and 38 men (17.3%) aged 21–33 years  of participants thought that information concerning meningitis risk and prevention was sufficiently accessible, but this opinion was less frequent among younger women (36.2%). The media , health care professionals and school teachers were mentioned most frequently as sources. In turn, health care professionals, the government, TV and radio were most frequently mentioned as sources that should provide information by those who considered that they did not receive enough. Younger women primarily expressed that health care professionals were in charge of providing this information.S2 Table). Half of participants reported that eating unripe mangos (49.3%) and dust (48.0%) were responsible for meningitis, according to the people in their circle. Only 5, (0.9%), mentioned person-to-person transmission. When asked about their own opinion on what caused meningitis, 58.7% mentioned dust, and 11.9% dust in the respiratory tract (Table 2). Among those citing dust, it came from wind (74.2%) or was inhaled (22.6%). Meningitis was associated with fire smoke for 1.9%, along with traffic pollution (1.3%) or unhygienic conditions (2.6%). Furthermore, 29.4% of participants thought themselves that meningitis was caused by eating unripe mangos and 2.8% mentioned contact between persons. Most participants gave only one factor, but a common combination was dust and eating unripe mangos. Younger women less frequently mentioned dust and more frequently did not know any causes.Frequently cited effects of dry air were meningitis (72.3%) and lower and upper respiratory infections  (Table 2). 2.3% mentioned avoiding eating unripe mangos and 1.4%, the application of shea butter in the nostrils. By contrast, when asked what they personally did to prevent from meningitis, only 25.9% mentioned vaccination, while half (45.9%) reported application of shea butter in the nostrils and 9.1% avoiding eating unripe mangos (Table 2). Vaccination was usually mentioned alone, but was sometimes combined with the application of shea butter in the nostrils, especially by older women. Avoiding dust exposure frequently was combined, especially with shea butter application in the nostrils, food hygiene and not eating unripe mangos. Surveying food hygiene, not eating unripe mangos or not knowing were more frequently mentioned by younger women (51%).To the question of how meningitis could be prevented, 84.5% of participants mentioned vaccination, followed by avoiding exposure to dust , and seeking professional help  (Most participants (90.6%) thought that children or “young people” (“les jeunes”) were at higher risk of meningitis, but explanations varied from higher vulnerability (rarely mentioned) to higher exposure to dust, smoke, eating unripe mangoes and poor food hygiene. Two participants mentioned gold miners and masons, due to their high dust exposure.Table 2).None of the participants declared having already refused “vaccination against meningitis” (no specific vaccine was highlighted in the question). However, seven (3%) said they knew someone who had refused, due to doubts about vaccine side effects (N = 3), lack of perception of its benefit (N = 2), fear of pain (N = 1) and lack of money (n = 1). Most participants declared making their vaccine decision themselves (68.6%), while parents decide for over one third of younger women and men. For 11.5% of women aged 21–33 years, the husband was indicated as the decision maker (S2 Table). Cough, sore eyes, cold or respiratory problems were mentioned each by about one third of participants as a consequence of kitchen fire smoke exposure, regardless of the age of the exposed person  (S3 Table). Most (81.2%) had heard of kitchen appliances that reduce smoke exposure, mainly gas stoves, “ROUMDE” and “Amelioré Foyer”. Most participants (95.9%) thought that children can be kept away from the smoke , while the lack of a person to mind the child and the child asking for their mother were mentioned as difficulties by eight participants (3.6%).Only 9 (4.1%) of participants mentioned dust or smoke in respiratory tract as a meningitis risk factor and only 3 (1.9%) of participants who had mentioned dust as a risk factor for meningitis mentioned fire and kitchen smoke as the origin of such dust (S1 Text). The score distribution was similar among the female (median 2) and male groups (median 3). However, younger women aged 15–20 years had higher a frequency of scores <2 (17.0%) than older women aged 21–33 years (6.3%) .The meningitis risk and prevention knowledge scores ranged from 0 (0.9%) to 4 (0.9%), with a median of 3 . Univariate associations were similar for achieving a high score (≥3), showing in addition an association with male gender. In parsimonious multivariate models, age ≥21 years  and at least one year of junior secondary school  both contributed significantly to having at least a basic score. Only secondary level education persisted as a determinant of a high score .In univariate analyses, achieving at least a basic score (≥2) was associated at P = 0.050 and older men OR 7.89, P = 0.062) (Table 3). A large part of the association was explained by at least one year of junior secondary school education , increasing the OR for older women to 5.53 (P = 0.006) and decreasing the OR among males. Specifically among younger women (M3), achieving at least a basic score was strongly associated only with education .In additional analyses including an age-gender term, achieving at least a basic score (≥2) tended to be substantially more likely (compared to younger women) among all other groups , but no substantial difference or trend was observed after adjusting for education, which was a significant determinant itself  (Table 3). Specifically among younger women (M3), achieving a high score was associated with both secondary level education  and the opinion of having sufficient information .Achieving a high score (≥3) was more likely among males  were known by the majority of participants, even though a gender gap which was largely explained by secondary school attendance, persisted among young women. Local understandings and practices for risk and prevention were commonly reported and used.Although respondents recognized the term “meningitis”, few people knew its biological etiologies, citing instead dust and dirt, often transported by wind, as well as consumption of unripe mangoes as causes. These responses were unsurprising in light of previous anthropological research in Africa and elsewhere around illness causation, and its relation to environmental and social factors, in particular malaria –21. In BHaemophilus influenzae type b (Hib), pneumococci and meningococci) and vaccines are not (sufficiently) available against all pathogens . These vaccines are routinely delivered only to children, although adolescents and young adults are also at risk. The link with extreme meteorological conditions is confirmed only for meningococcus and pneumococcus, and epidemics are nearly always due to meningococcus. Popular knowledge regarding the disease known under terms such as “stiff neck” therefore may mainly apply to meningococcal and pneumococcal meningitis.The reported local understandings of meningitis risk and prevention must be interpreted in light of complex biomedical explanations and complex public health measures. Meningitis, defined for public health purposes as acute bacterial meningitis, has various etiologies and occurs in various age groups, but meningitis in infants usually shows unspecific clinical presentation. Meningitis vaccine prevention is complex, as it requires several vaccines  from entering the bodies and making them ill . For exaThe link between smoke exposure and pneumonia is well established , 26, andStudy participants rarely mentioned interpersonal disease transmission, despite its etiological importance for an infectious disease such as meningitis. In Hayden et al.  only halThe educational gender gap has been targeted by organizations such as UNICEF over decades. Less is known about a gender gap for health literacy in sub-Saharan Africa, in particular infectious disease prevention apart from sexually transmittable diseases. One study in Ghana recently described a similar gender gap, in part related to educational differences, and suggested that “dominant male gender roles and differing expectations and opportunities for socio-economic advancement for men and women” may lead to the observed differences . ClosingThe additional difference between women below and above age 21, which persisted after adjustment for education, can be interpreted in at least two ways: firstly, women aged ≥21 years were old enough in 2010 to understand and retain the information provided during the MenAfriVac communication campaign, whereas the younger age group (<15 years in 2011) did not benefit from this information campaign even if they were vaccinated. This interpretation would suggest that such campaigns have long-term benefits for knowledge transmission and retention and should be repeated periodically to catch-up with younger cohorts. A second interpretation is that women may receive health information primarily from health professionals during perinatal care, while men receive it through various other channels. Women under 21 years of age in this urban population would have been less exposed to educational information from health professionals, in the absence of an early pregnancy. This interpretation implies that for infectious diseases, including sexually transmittable diseases and human papilloma virus, where infection can occur before young women and adolescents have consistent access to health care professionals, specific educational interventions are needed outside school and health care facilities.More than half of study participants—and two thirds of young women—claimed that they did not receive sufficient information about meningitis prevention; most expected this information to come from health professionals and the media. There is a crucial role to be played by public health authorities, health workers and the media in improving health literacy and promoting vaccine uptake. Such informational transfers can take place in many forms, from exchanges at health centers to radio announcements, but should, as pointed out above, include settings frequented by young women who are no longer at school and not yet seen in perinatal care. Interventions should be consistent and respectful of local explanations and logics around meningitis risk and prevention. Eighty-two percent of respondents mentioned vaccination as a means of prevention, which suggests that public health messaging around meningitis prevention does reach populations. Participants rarely reported vaccine refusal by themselves or people they know; when they did, participants reported reasons associated with vaccine hesitancy as defined by WHO : concernOur study has several limitations. By nature, information was self-reported and may be subject to social desirability bias, as participants may have perceived pressure from the interviewing nurse or from their social environment regarding expected beliefs. We sought to reduce this bias by investigating participants’ and their networks’ perceptions and practices: What do others think? What do you think? What do you do?Our results are limited to an urban and peri-urban population and cannot be extrapolated to a rural population, where the balance between access to information and the importance of local beliefs may be different. Ethnic specificities may have influenced our results; however, Bobo-Dioulasso is an economically significant town and its inhabitants come from various regions of Burkina Faso and West Africa for trade and higher studies, such that it is unlikely that our results are dominated by one single ethnicity.Our definition of the knowledge score was based on a binary categorization of responses as “correct” and “incorrect” and it included some questions to which even health care workers may have found difficult to reply to correctly. In addition, for some questions, several responses were possible. A person could get one point for giving an expected response, while also citing other incorrect aspects. Finally, the sample size was not sufficient for some stratifying analyses on gender, age and socioeconomic status.At the same time, however, our methodology, which combines a knowledge, attitude, beliefs and practice (KABP) study with a qualitative component, demonstrates the usefulness of this type of mixed approach. KABP studies have been roundly criticized for putting complex perceptions into simplistic boxes, but remain in widespread use. By adding qualitative, open-ended questions, we were able to gain access to rich explanatory data on practices and understandings related to risk and prevention of meningitis that we otherwise would not have collected.In conclusion, our study shows a gender gap in knowledge about meningitis risk and prevention, largely due to education inequalities in Burkina Faso. Women aged 20 or younger had particularly low levels of knowledge and should be targeted by interventions outside schools and perinatal care. Our study suggests a strong adherence among young adults to local understandings of and practices around meningitis risk and prevention. National and international vaccination initiatives need to take these local understandings and practices into account.S1 Text(DOC)Click here for additional data file.S1 TableOriginal in French and authors’ translation into English.(PDF)Click here for additional data file.S2 TableParticipants could provide more than one theme.(PDF)Click here for additional data file.S3 TableParticipants could provide more than one theme.(PDF)Click here for additional data file.S4 TableEstimates were obtained from bivariate logistic regression models.(PDF)Click here for additional data file.S1 Dataset(DTA)Click here for additional data file.


Diabetic kidney disease (DKD) is a kind of common microvascular complication of diabetes. This study aims to explore the possible links between blood sugar level and albuminuria, providing the exact cut point of the “risk threshold” for blood glucose with DKD.The relationship between blood glucose and albuminuria was modeled using linear and logistic regression in the REACTION study cohorts (N= 8932). Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by logistic regression model. Two-slope linear regression was used to simulate associations between blood glucose and ACR.We found that the increase in ACR was accompanied by increased HbA1c, with a turning point at 5.5%. The positive correlation remained highly significant (P<0.001) when adjusted for age, sex, marital status, education, smoking status, drinking status, BMI, waistline, SBP and DBP. In subgroup analyses including gender, obesity, hypertension, and smoking habits, the relationship was significant and stable.We determined a risk threshold for HbA1c associated with albuminuria in a Chinese population over the age of 40. HbA1c ≥ 5.5% was positively and independently associated with ACR. These results suggest the necessity of early blood glucose control and renal function screening for DKD in at-risk populations. Diabetic kidney disease (DKD) is the most common microvascular complication of diabetes and the most common cause of end-stage renal disease (ESRD) . About 3Hemoglobin A1c (HbA1c) represents the average fluctuation level of blood glucose in the body in recent three months. As a significant change in clinical practice, the American Diabetes Association’s 2010 guidelines recommended HbA1c as one of the diagnostic criteria for diabetes, and set a threshold of ≥ 6.5% . Since tThe objective of our research is to explore the correlation between blood glucose and the continuous increase in ACR values and to reveal the adverse effects of hyperglycemia on renal function prior to the diagnostic criteria for diabetes being met. We attempt to determine the optimal blood glucose threshold to provide additional clinical evidence for the prevention and early screening of diabetic kidney injury, so as to prevent the occurrence and development of DKD.We conducted a cross-sectional study of community populations from March to June 2011. Our research population was obtained from the longitudinal REACTION study, which aimed to assess cancer risk and related risk factors in Chinese diabetic population. Specific information regarding experimental design and cohort recruitment has been previously reported , 23. OriWe designed a questionnaire survey to gather information on demographic data  and living habits (smoking and drinking). Education level was categorized as elementary school or below, junior high school, technical secondary or high school, and college degree or above. Smoking and drinking habits were defined as never, former, or current, indicating who had regularly smoked or consumed alcohol during the past 6 months.Participants received a physical examination. Their height and weight were measured in units of 0.1cm and 0.1kg, and BMI (kg/m2) was calculated. Waistline was defined as the horizontal circumference through the umbilical center measured with a soft tape measure, at the end of exhalation and before the beginning of inhalation. Hipline was measured at the most convex region of the pubic symphysis and the gluteus maximus, with the legs close together and the arms resting naturally at the sides. Waist-to-hip ratio (WHR) was calculated as waistline divided by Hipline. Blood pressure was measured after at least 5 minutes of rest, and the average of three measurements was taken .2 as the unit and used the following formula: eGFR = 175 × [serum creatinine × 0.011]-1.234 × [age]-0.179 × [0.79 if female] . The level of HbA1c was detected by high-performance liquid chromatography . For the OGTT test, blood glucose levels were measured 0h before and 2h after 75g glucose orally. We collected the first morning urine sample of participants for laboratory testing. Urinary albumin levels in the samples were determined by chemiluminescence immunoassay . And creatinine levels were determined using Jaffe kinetic method , respectively. ACR was estimated as the urine albumin to creatinine ratio and is expressed as mg/g. eGFR took mL/min per 1.73 m female] .2 or the presence of persistent severely elevated albuminuria  of >30 mg/g)  criteria, including fasting blood glucose greater than or equal to 7.0 and/or OGTT 2 h greater than or equal to 11.1, or self-reported history of diabetes . We incl30 mg/g) .post hoc comparisons. Continuous variables with nonnormally distributed data are indicated as median and interquartile range (IQR), and Kruskal-Wallis test was performed to compare group differences. Categorical variables are presented as numbers (proportions), and differences between groups were performed with the χ2 test. The unadjusted and multivariate-adjusted linear and logistic regression analysis, calculating odds ratios (ORs) and corresponding 95% confidence intervals (95% CI), were used to identify risk factors for increased risk for diabetes. A 2-slope linear regression was used to model associations between HbA1c and ACR. Adjustment was then made a priori for relevant covariate: age, sex, marital status, education, smoking status, drinking status, BMI, waistline, systolic blood pressure, diastolic blood pressure, fasting plasma glucose, oral glucose tolerance test and HbA1c. To determine whether eGFR CKD, hypertension and central obesity interact with HbA1c increments in predicting ACR, product terms for (eGFR ≥ 60) × HbA1c, CKD × HbA1c, hypertension × HbA1c, central obesity × HbA1c were added into the regression model with covariate adjustments as mentioned above. All statistical analyses were performed using RStudio version 3.6.1. A two tailed p<0.05 was considered to be statistically significant.Continuous variables that satisfy a normal distribution are presented as the mean ± standard deviation (SD), one-way ANOVA was used to analyze group differences, and Bonferroni correction was performed for This research finished without patient involvement. Patients were not invited to participate in research design, data analysis and manuscript writing.A total of 8,932 participants completed the collection of demographic, physical examination, and laboratory data. Clinical characteristics of the participants are shown according to ACR category grouped by quartile in ACR was positively skewed and was logarithmically transformed before linear regression analysis. The relationships between blood glucose and log-ACR were examined using 2-slope linear regression models. As shown in We next explored the association between glucose levels and ACR in continuous using linear regression analyses, as well as ACR ≥ 30 in category using logistic regression analysis in To examine the independent effects of eGFR, CKD, hypertension, central obesity on the relationship between HbA1c and ACR, interaction terms were introduced into the linear regression model. Back-transformed coefficients from this model are presented in The subgroup analyses took into account gender, obesity, hypertension, and smoking habits, and the positive relationship between HbA1c and ACR remained stable and significant. This positive correlation was not affected by central obesity, hypertension or gender Figure 2Our study provides evidence that among blood glucose indicators, only HbA1c is independently associated with ACR. The risk threshold identified for HbA1c for the prevalence of albuminuria among a Chinese population was 5.5%. The increase in ACR was accompanied by a positive change in HbA1c, when HbA1c ≥ 5.5%. Furthermore, the correlation remained stable and significant after adjusting for sociodemographic characteristics and performing subgroup analyses based on gender, smoking habit, obesity, and hypertension. This result indicates the effect of blood sugar on renal damage may occur before the diagnosis of diabetes, emphasizing the necessity of early blood glucose control and renal function screening for DKD.At present, many studies have shown that HbA1c levels affect albuminuria. For people with diabetes, data from the FinnDiane study and DMIDS project both suggest that HbA1c influences the development of albuminuria and renal disease , 30. MeaWith respect to a glucose threshold in terms of HbA1c levels, current clinical studies have not consistently demonstrated a relationship for predicting the onset of DKD. In the diabetic population, the ADVANCE study showed no significant increase in the risk of retinopathy or renal complications with an HbA1c level below 6.5% (48 mmol/mol) . This reCurrently, there is sufficient evidence that intensive glucose therapy significantly reduces albuminuria and improves the composite end point of nephropathy. As of 2019, the ADA recommended target for T2DM glycemic control is HbA1c < 6.5% after adequate consideration of the risk of hypoglycemia. The DCCT study of a T1DM population, the UKPDS study of a T2DM population, and the ADVANCE study all showed that intensive glycemic therapy significantly improved renal clinical benefits compared to conventional glycemic control , 47. TheThe National Institute for Clinical Excellence (UK) and the Standards of Medical Care in Diabetes from ADA both recommend kidney care for all people with type 2 diabetes that includes measuring ACR or albumin levels annually , 49. BasOur analysis had the following limitations. First, the design of this study was a cross-sectional survey, which can only identify the correlation but cannot conclude cause and effect between ACR and HbA1c. Second, although we used two regression models to estimate a single point of change, the choice of the point of change may also be skewed. However, this deviation, while having a slight effect on the regression coefficient, does not affect the nonlinear conclusion. Furthermore, while we adjusted for many of the covariables associated with ACR in the multiple regression analysis, other potential factors, such as social status, personal income level, and family lifestyle may also influence the regression results and should be included in the adjustment. Third, we only used morning urine samples to measure ACR levels, but levels of ACR fluctuate with time. Therefore, combining this measurement with 24-hour urine albumin quantitative results to assess the risk of albuminuria would be more scientific and rigorous. However, there was a good correlation between on-site urine ACR samples and urine samples collected 24 hours a day. Urine ACR assessment through field samples may be a reliable alternative to epidemiological specimen collection . Fourth,Our study provides evidence for an HbA1c threshold in which the risk of albuminuria begins to increase at a cutoff point of 5.5% in a Chinese population. We found that HbA1c≥5.5% was positively and independently associated with ACR. After adjusting for the influence of risk factors, the correlation remained stable and significant. This finding underlines the need for screening renal function in at-risk populations and the importance of early glucose management to delay the development of renal complications from diabetes.xumt@mail.sysu.edu.cn with proper purposes.Data are available upon reasonable request. Main document data and additional unpublished data from the study are available by sending Email to The studies involving human participants were reviewed and approved by Ruijin Hospital Ethics Committee Shanghai JiaoTong University School of Medicine. The patients/participants provided their written informed consent to participate in this study.All authors contributed to the conception of the study, execution of data collection, data analysis or writing and revision of the paper. All authors contributed to the article and approved the submitted version.This work was supported by National Key R & D Program of China (No. 2016YFC0901204), Medical Scientific Research Foundation of Guangdong Province of China (A2019040), and National Natural Science Foundation of Guangdong, China (2019A1515011110).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.


Lavandula angustifolia L. (lavender), named LA 2019 and LA 2020, respectively, grown in central Italy was analyzed and compared by GC and GC-MS. For both samples, 61 compounds were identified, corresponding to 97.9% and 98.1% of the total essential oils. Explorative data analysis, performed to compare the statistical composition of the samples, resulted in a high level of global similarity (around 93%). The compositions of both samples were characterized by 10 major compounds, with a predominance of Linalool (35.3–36.0%), Borneol (15.6–19.4%) and 1,8-Cineole (11.0–9.0%). The in vitro antibacterial activity assay by disk diffusion tests against Bacillus subtilis PY79 and Escherichia coli DH5α showed inhibition of growth in both indicator strains. In addition, plate counts revealed a bactericidal effect on E. coli, which was particularly noticeable when using oil from the fresh lavender flowers at the highest concentrations. An in vitro antifungal assay showed that the EOs inhibited the growth of Sclerotium rolfsii, a phytopathogenic fungus that causes post-harvest diseases in many fruits and vegetables. The antioxidant activity was also assessed using the ABTS free radical scavenging assay, which showed a different antioxidant activity in both EOs. In addition, the potential application of EOs as a green method to control biodeterioration phenomena on an artistic wood painting (XIX century) was evaluated.The chemical composition of essential oils (EOs) from dried and fresh flowers of  Among the most investigated natural sources of secondary metabolites, aromatic plants play a fundamental role due to their essential oils (EOs) ,2,3 and,Lavandula genus (family Lamiaceae), a small, fragrant shrub native to the Mediterranean area, is cultivated worldwide for its essential oils, which find applications in various industrial areas, such as perfumes, pharmaceutics and cosmetics; furthermore, lavender essential oil is widely used in the food industry because of its biological properties as well as its attractive aroma  × 100, where Abs control is the absorbance of the ABTS•+ + methanol; Abs sample is the absorbance of ABTS•+ + EO. Ascorbic acid was used as a standard.The ABTS method was used based on the ability of antioxidant molecules to quench the long-lived ABTS changes . BrieflyMadonna con Bambino”, originally located on the inner wall of the S. Maria del Lago church at Pesche , A historical wood painting was chosen for the ex situ study “Microbiological analyses were carried out as follows: (i) sampling by soft and non-destructive techniques using sterile cotton swabs and sterile velvet tampons b; (ii) tThe antimicrobial activities (antagonistic properties) of the isolated bacteria were evaluated in vitro using the EO diffusion agarized nutrient method. Only the yellow-pigmented bacterial strains typically representative of biological particulates both in the air environment and the surface of the potential biodeteriorated paint were identified. The tests included morphological properties under microscopic observations and physiological and biochemical tests by a Gram reaction and Api-Systems  according to Bergey’s Manual .L. angustifolia EOs were tested using both agar wells and disk diffusion on Petri dishes containing Plate Count Agar  and inoculation of selected strains. The suspensions containing exponentially growing bacteria after an overnight broth culture, at approximately 108 CFU / mL, corresponding to an O.D. 560 nm of approximately 1.6, were used [For the assay against yellow-pigmented isolated bacteria, the ere used ,62.Paper disks with EO microdilution (10 μL/disk) were carried out in triplicate. As a control, a sterile physiological solution instead of EO dilutions were adopted. The plates were incubated at 37 °C for 48–72 h, and the halos of inhibition around the disks were measured (in mm).L. angustifolia and the antimicrobial and antioxidant activity was observed. The two samples (LA 2019 and LA 2020) were qualitatively similar and contained Borneol and Linalool in high concentrations. This is the first time that an EO from L. angustifolia with a high Borneol content has been described in Italy. The differences observed between the two samples (dried and fresh lavender) concerned only the concentration of Borneol, Linalool and 1,8-Cineole, suggesting an almost total conservation of the components during the drying process. In our experimental conditions, we presumed that the antibacterial and antifungal activity was partially influenced by the concentration of these components.In the present study, a relationship between the composition of fresh and dried EOs of flowers extracted from B. subtilis PY79 and E. coli DH5α, with a significant bactericidal effect on the latter as well as an antifungal effect on the phytopathogen fungus S. rolfisii. These activities are associated with a scavenging capacity; therefore, our results suggest that the EO can be used as potential biopreservatives in several fields by reducing and inhibiting pathogenic bacteria. Their antioxidant activity makes L. angustifolia EOs a favorable candidate for decreasing the level of microbial contamination not only in different categories such as cosmetic products  but also in potential “natural” alternatives to synthetic antioxidants/bactericides.The Borneol-rich oils harvested in central Italy were found to have antibacterial activity against both The interesting antimicrobial activities, if confirmed with further studies on the biodeteriogen microflora associated with the artworks, could be useful in establishing a new green biological strategy for the recovery of altered artworks and as an alternative to the use of biocides and toxic compounds.


About 20–40 % of autistic people experience a phenomenon of regression. Retinol binding protein 4 (RBP4) plays an important role as an inflammatory neurotrophic adipokine and is a promising mediator of the fat-brain axis. Abnormal fatty acid metabolism and lipid mediators have been reported to be related to the etiological mechanism in autism, and amelioration of impaired lipid metabolism can be recognized as a treatment strategy for autism. The purpose of this study is to explore the relationship between RBP4, lipids, and the autistic regression phenomenon, and to discuss their potentials as biomarkers for the autistic regression phenomenon.A total of 60 autistic individuals  (ASD group) and 36 healthy controls were enrolled in this case-control study. The levels of RBP4, total cholesterol (TC), high-density lipoprotein (HDLC), low–density lipoprotein (LDLC), and triglyceride (TG) were measured. Childhood Autism Rating Scale (CARS) is used to assess the severity of autism. Ethical measures were performed in compliance with the current Declaration of Helsinki and written informed consent was obtained from the parents before enrollment of the children and adolescents.P = 0.007), RBP4 (P = 0.001), and HDLC (P = 0.027). The levels of RBP4 in ASD group were positively correlated with TG , HDLC , TG/TC  and TG/LDLC , and were negatively correlated with CARS . Further logistic regression demonstrated that decreased RBP4 concentration was associated with the presentation of the autistic regression phenomenon even after the adjustment of the potential confounding factors.Compared with control subjects, autistic individuals had lower levels of TC (Serum RBP4 is associated with the autistic regression phenomenon and the severity of ASD. Further studies are needed to expound whether decreased RBP4 participates in the development of the autistic regression phenomenon. Autism spectrum disorder (ASD) is mainly manifested in the deficits of social communication and social interaction, as well as repetitive behavior, limited or restricted interest or activity . EpidemiThe exact etiology and mechanism of the autistic regression phenomenon remain unclear. Current research assumes that it may be related to the interaction between biological and environmental factors , 11, 12.Former studies have identified that plasma RBP4 levels both in the first trimester and second trimester are dose-dependently associated with increased risk of gestational diabetes mellitus (GDM) . In addiAdditionally, abnormal fatty acid metabolism and lipid mediators have been reported to be related to the etiological mechanism in autism, and amelioration of impaired lipid metabolism can be recognized as a treatment strategy of autism –21. FurtAutistic participants were 60 children and adolescents aged 3 to 14 years old, 39 males and 21females, the average age was 7.3 ± 3.2 years. Healthy control participants were 36 children and adolescents aged 3 to 14 years old, 12 females and 24males, the average age was 7.3 ± 3.1 years. Autistic participants were recruited from the outpatient department of child and adolescent psychiatry in Shanghai Mental Health Center between September 2018 and August 2019. All autistic participants were by the diagnostic criteria of autism spectrum disorders proposed by the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5). Healthy controls were recruited from the children who came to the same hospital during the same period for regular consultation but had not been diagnosed with any mental disorders. Individuals with Fragile X syndrome or other pervasive developmental disorders, obsessive-compulsive disorder, affective disorder or other neuropsychiatric disorders, neurodegenerative disorders, brain trauma or cerebrovascular disorders, metabolic disorders, or immunological disorders were excluded.This study was performed in strict accordance with the Declaration of Helsinki and other relevant national and international regulations. All procedures for this study were approved by the institutional review boards of Shanghai Mental Health Center. Written informed consent was obtained from the parents before enrollment of the children and adolescents.Child Autism Rating Scale (CARS) was applied to evaluate autistic symptoms . Total sAutistic participants were further assessed based on the following questionnaire  and wereThe venous blood was collected and the serum and cellular components were separated within 2 h after 3000 g centrifugation for 10 m. The obtained supernatant (serum) was stored at -80 °C for further analysis. The levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured by the enzymatic method on Olympus AU5400 automatic biochemical analyzer .Serum RBP4 levels were assayed in duplicate by using a sandwich enzyme-linked immunosorbent assay (ELISA) kit  according to the manufacturer’s protocol.2 test was used to compare the gender between the two groups. Mann-Whitney U test was used for statistical comparison of continuous data without normal distribution. Spearman’s rho test was used to analyze the correlations with the data of nonnormal distribution. Normally distributed data were tested using Pearson’s correlation. Logistic regression was used to analyze the risk factors for the autistic regression phenomenon, and the odds ratio (OR) and 95 % confidence interval (CI) were calculated. Crude OR and adjusted OR were estimated by bivariate and multivariate logistic regression analysis, and the crude model analyzed the crude data only, the adjusted model was adjusted for age, gender, BMI, TC, TG, LDL-C, HDL-C, CARS. Receiver operating characteristic (ROC) curve analysis was used to determine the optimum cut-off levels of RBP4 and TG to predict the autistic regression phenomenon. All statistical tests were two-tailed, and p < 0.05 was considered statistically significant.IBM SPSS Statistics was used for data analysis. The normality of data distribution was determined with the Shapiro–Wilk test. Normally distributed data were expressed as (mean ± standard deviations). The skewness distributed variables were reported as the median (interquartile range). An independent sample t-test was used to compare the normally distributed continuous data between the two groups. Χ P < 0.01), and a lower level of HDLC (P = 0.027).The demographic data and biochemical characteristics are presented in Table P < 0.01).Sixty autistic participants were subdivided into 18 participants with regression phenomenon and 42 individuals without regression phenomenon. Compared with those without regression phenomenon, individuals with regression phenomenon had lower TG and RBP4  and TG/LDHC , were negatively correlated with CARS . In ASD-NR group, the levels of RBP4 were also positively correlated with TG  and TG/LDHC , were negatively correlated with CARS . In the whole ASD group, the levels of RBP4 were positively correlated with TG , HDLC , TG/TC , TG/LDLC , were negatively correlated with CARS .The correlation of RBP4 with clinical parameters in autistic participants are shown in Table P = 0.005). In addition, the optimal cut-off value of RBP4 for the prediction of the autistic regression phenomenon was 20.6 ug/ml, with a sensitivity of 94.4 % and a specificity of 42.9 % .As shown in Fig. After the adjustment of the potential confounding factors including age, gender, BMI, CARS, and the levels of TG, TC, HDL-C, LDL-C, logistic regression demonstrated that decreased RBP4 concentration increased the risk of the autistic regression phenomenon  Table .The present study demonstrated that autistic participants had lower levels of TC, RBP4, and HDLC. Additionally, autistic participants with regression phenomenon had lower levels of TG and RBP4. Furthermore, the levels of RBP4 were positively correlated with TG and were negatively correlated with the severity of autism. Moreover, decreased RBP4 concentration increased the risk of the autistic regression phenomenon.Consistent with the present study, some researchers also found that there were significant differences in TG, TC, HDL-C, non-HDL-C, and TC/HDL ratio in ASD group and healthy volunteers regardless of their diets , 26. DifWhen compared with those without regression phenomenon, the present study demonstrated that autistic participants with regression phenomenon had lower TG. However, the present study didn’t detect the significant difference of TC or HDLC between ASD-R and ASD-NR groups. This indicates that TG may be related to the autistic regression phenomenon, whereas TC and TG may play different roles in the progression of the autistic regression phenomenon. Similarly, some researchers have found that mainly amino acid, lipid, and nicotinamide metabolism were significantly different between ASD with regression phenomenon group and ASD without regression phenomenon group . In geneFormer studies have found that Serum RBP4 levels of major depressive depression individuals were significantly lower than that of the healthy control group . HoweverThe results provide evidence that lipids and RBP4 were related to the presence and regression phenomenon of ASD. They can jointly predict the early diagnosis of ASD, guide early clinical intervention, early identification of regression phenomenon, and evaluate the prognosis of ASD.The present study observed a significant difference in the serum level of RBP4 in ASD and healthy controls for the first time as well as the correlation between RBP4 and lipids. Nevertheless, several limitations of the present study should also be considered. As the study was performed in the Chinese Han population, the results need to be confirmed in other regions and ethnicities. In addition, the relatively small sample size may cause a potential low power of our results. Besides, the age range of the subjects included in this study was 3 to 14, whether those autistic participants who are out of this age range have similar characteristics is not clear. Further study is needed to expand the age range to demonstrate the changes in lipid and RBP4 throughout the life cycle of autistic participants.In summary, autistic participants had lower levels of RBP4, TC, and HDLC. Furthermore, compared with those autistic participants without regression phenomenon, autistic participants with regression phenomenon had lower TG and RBP4. Meanwhile, the levels of RBP4 were positively correlated with TG and were negatively correlated with CARS. Moreover, decreased RBP4 concentration increased the risk of the autistic regression phenomenon. Altogether, these results suggest that the serum RBP4 in combination with lipids may be considered predictors associated with the autistic regression phenomenon. Further studies are needed to expound whether decreased RBP4 participates in the development of the autistic regression phenomenon. The results indicate that correcting lipids and RBP4 to appropriate levels may help clinicians and practitioners in early intervention of the autistic regression phenomenon. However, it is important to note that the results must be interpreted with caution. Due to the heterogeneity of the condition, as well as the huge difference in developmental trajectories, the intervention should be tailored to that particular child regardless of being in a possible regression-risk group or not.


Xenograpsus testudinatus and the sediments of its habitat. The highest number of colonies was obtained from Zobell marine agar plates with an overall number of 29 genetically distinct isolates. 16S RNA gene sequencing using the Sanger sequencing method revealed that most of the bacterial species belonged to the phylum Firmicutes and the class Bacilli. The present study indicates that hydrothermal vent bacteria and their secondary metabolites may play an important role for the reconstruction of the evolutionary history of the phylum Procaryota.Extreme environments are hostile for most organisms, but such habitats represent suitable settings to be inhabited by specialized microorganisms. A marine shallow-water hydrothermal vent field is located offshore in northeast Taiwan, near the shallow shore of the southeast of Kueishantao Island . Research on extremophilic microorganisms makes use of the biotechnological potential associated with such microorganisms and their cellular products. With the notion that extremophiles are capable of surviving in extreme environments, it is assumed that their metabolites are adapted to function optimally under such conditions. As extremophiles, they need specific culture conditions, and only a fraction of species from the original samples are recovered in culture. We used different non-selective and selective media to isolate bacterial species associated with the hydrothermal vent crab  This compound showed significant inhibition against multidrug-resistant Staphylococcus aureus with an MBC of 20 ± 0.072 mg L−1 and an MIC of 15 ± 0.172 mg L−1. The compound showed high antioxidant activity and was non-hemolytic. Saggese et al. [Bacillus pumilus (SF214) produced a protein compound with antibacterial activity. The small molecule of 3 kDa exhibited activity against Staphylococcus aureus and the other larger molecule of 10 kDa was active against the pathogen Listeria monocytogenes. A mass spectrometry analysis classified the small molecule as pumilacidin, a non-ribosomally synthesized biosurfactant that is active against Staphylococcus. Nisha et al. [Micrococcus luteus isolated from a ship hull at Cochin port, India. An EPS-associated compound from M. luteus was isolated and characterized, and the results showed antibacterial and antifungal activities against pathogens. An in vitro assay for cytotoxic activity of the pigment indicated antibacterial and antifungal activity. Furthermore, UVA rays’ potential to absorb the pigment makes it unstable when applied in sunscreens.He et al.  used pren et al.  isolatede et al.  stated ta et al.  characteXenograpsus testudinatus were Kocuria subflava, Paenibacillus silvae, Bacillus tequilensis and Micrococcus luteus; those restricted to sediments were Bacillus subterraneus, Brevibacillus parabrevis, Bacillus marisflavis, Bacillus velezensis, Jeotgalicoccus huakuii and Bacillus megaterium; those restricted to the ventral crab plate were Staphylococcus sciuri, Bacillus flexus, Bacillus halosaccharovorans, Bacillus jeotgali, Bacillus licheniformis, Bacillus amyloliquefaciens and Staphylococcus haemolyticus; and those from the carapace  were Bacillus firmus, Bacillus safensis, Bacillus iocasae, Bacillus aquimaris, Bacillus marisflavis, Bacillus vallismortis, Exiguobacterium mexicanum and Psychrobacter pulmonis. The species common to the gut and sediment were Bacillus ginseng and Bacillus aryabhattai. The bacteria found both in the sediments and on the carapace were Exiguobacterium aurantiacum, Bacillus cereus and Bacillus albus. Bacillus aerius was isolated from both the ventral plate and the carapace  and sediment samples from Kueishantao hydrothermal vents were aseptically collected and immediately processed after bringing them to the laboratory. Different media were prepared to isolate a maximum number of individual bacterial species. The media used were Nutrient agar, Luria Bertani agar, Tryptic soy agar, TCBS media, Zobell marine media, brain heart infusion agar, sea water complete and Casamino acid sea water. All the media were prepared in either sea water or distilled water. From the crab samples, swabs were prepared from the crab dorsal, ventral portion and gut and were spread on all the respective media plates. The crab samples were homogenized and serially diluted up to 105. The sediment samples were serially diluted to 105, and 100 µL from each dilution was spread on all the different media plates. The plates were incubated at 27 °C, 32 °C, 37 °C and 42 °C.Crab A single colony was inoculated in the respective medium, which showed the optimal growth of the bacteria at the optimal temperature and was kept in a shaking incubator at 150 rpm. Genomic DNA was isolated applying a GeneJET Genomic DNA Purification Kit . The PCR reaction with Taq buffer (1X), dNTP (0.2 mM), reverse and forward primers (1 µM each), 50–100 ng template DNA, Taq DNA polymerase (1.25 U) (New England biolabs) and bi-distilled water to a final volume of 25 µL was set. Universal primers, 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′-GGTTACCTTGTTACGACTT-3′) were used to amplify the 16S rRNA sequence. The PCR reaction conditions were set with initial denaturation of 94 °C for 5 min, 35 cycles of 95 °C (1 min), 55 °C (1 min), 72 °C (1:40 min) and final elongation at 72 °C (10 min), and the amplified PCR product was run in agarose gel and purified by a GeneJet PCR purification kit . The eluted DNA samples were sent for 16S rRNA sequencing.For 16S rRNA sequencing, both forward (27F) and reverse (1492R) primers were used. Sequences were obtained by Sanger sequencing. Forward and reverse sequences were analyzed and assembled using MEGAX . The ful4. Different concentrations of ZnSO4 were used to study its effect on protease activity.A protease assay was performed using skim milk powder. Skim milk agar plates were prepared using 1% skim milk powder. The bacterial colonies were inoculated in broth and allowed to grow overnight. The overnight culture was streaked on pre-prepared agar plates and incubated at 37 °C again overnight. A zone of lysis was observed. A study was performed with and without the presence of ZnSOBacillus. The number of bacterial colonies obtained from the carapace was higher than compared to other sources. Bacterial colonies differed according to sample origin. The highest number of colonies was isolated from HV crab carapace and sediments. This study was carried out to derive bacteria from extreme conditions with the ability to tolerate a wide range of pH, high temperatures, varying pressure and nutrient availability. This provides the expectation of extracting unique metabolites that protect organisms from extreme physicochemical factors. Additionally, this can be used to study the pathways adopted by such extremophiles to adapt to toxic compounds and physical conditions in such extreme environments. Microbial adaptations at the genomic, proteomic and metabolomic level are expected to be of substantial applied interest for biotechnology, medical and industrial applications.Using different cultivation methods, a maximum number of colonies were isolated, and distinct colonies were classically identified by different morphology or color. The present study is the first report of microbial diversity derived from laboratory cultures that were classified by sequencing. The sequencing results revealed that most of the bacterial species belonged to the phylum Firmicutes. The classified bacteria mostly belonged to the genus


Parus major). Using an Illumina metabarcoding approach which amplifies the 16S bacterial ribosomal RNA gene, we measured gut microbiota diversity and composition from 49 great tit nestlings, originating from 23 different nests that were located in 22 different study plots across a gradient of forest fragmentation and tree species diversity. Per nest, an average microbiome was determined on which the influence of tree species (composition and richness) and forest fragmentation (fragment area and edge density) was examined and whether this was linked to host characteristics (body condition and fledging success). We found an interaction effect of edge density with tree species richness or composition on both the microbial richness  and community structure (beta diversity: weighted and unweighted UniFrac). No significant short-term impact was observed of the overall faecal microbiome on host characteristics, but rather an adverse effect of specific bacterial genera on fledging success. These results highlight the influence of environmental factors on the microbial richness as well as the phylogenetic diversity during a life stage where the birds’ microbiota is shaped, which could lead to long-term consequences for host fitness.Despite the microbiome’s key role in health and fitness, little is known about the environmental factors shaping the gut microbiome of wild birds. With habitat fragmentation being recognised as a major threat to biological diversity, we here determined how forest structure influences the bacterial species richness and diversity of wild great tit nestlings ( During the past two decades, the gut microbiome has attracted considerable research attention because of its role in a host’s physiology and health status. The microbiota of the gastrointestinal tract plays a fundamental role in food digestion, pathogen defence, stimulation of the immune system, gut and central nervous system functioning, life-history traits and behaviour . DespiteInitially the gut microbiota is shaped by external bacteria present in the nesting environment and by parental feeding . Food-asParus major) were shown to feed their nestlings less caterpillars in small fragments compared to larger fragments  and forest fragmentation (fragment area and edge density) on the gut microbiota of wild birds. We use great tits, an insectivorous forest bird during breeding season, as a model species by sampling the gut microbiota in 49 great tit nestlings originating from 23 different nests to determine the average nest microbiome. The nests were located in 22 different forest fragments in the south of Ghent, northern Belgium, with a varying fragmentation gradient and tree species composition. Taking into account both forest composition and forest fragmentation allowed us to address the question whether alpha and beta diversity are shaped by forest structure and whether it has consequences for bird health and fitness. Previously, it has been shown that body condition of great tits is an important factor for bird fitness, especially for bird survival after fledgling . With thQ. robur), Red oak (Quercus rubra), and Beech (F. sylvatica) in monocultures, two species mixtures or three species mixtures] (We performed a gut microbiota study of 23 great tit nests in 22 study plots (30 × 30 m) located in 11 mature (>60 years) deciduous forest fragments in the south of Ghent , northern Belgium . Plots vixtures] . Plots oAs explained in n = 49) were sequenced and analysed on nest level (n = 23).For full details on the field work procedures we refer to n = 49) using the CTAB method modified from 2HPO4) and 0.5 ml phenol-chloroform-isoamyl alcohol (25:24:1). The mixture was homogenised by grinding (2×) with 0.5 g unwashed glass beads  in a bead beater  with a 30 s interval between shakings. After centrifugation , 300 μl of the supernatant was transferred to a new tube. A re-extraction from the remaining content was performed by adding 0.25 mL CTAB buffer. After homogenisation as described above, samples were centrifuged  and 300 μl of supernatant was added to the first 300 μl. An equal volume (600 μL) of chloroform-isoamyl alcohol (24:1) was added to the supernatant collected in order to remove the phenol from the samples. The mixture was further centrifuged at 16 000 g for 10 s and the aqueous phase was transferred to a new Eppendorf tube. Nucleic acids were precipitated with two volumes of PEG-6000 solution  for 2 h at room temperature. After centrifugation , the pellet was rinsed with 1 ml of ice-cold, 70% (v/v) ethanol. The pellet was dried and resuspended in 100 μl RNAse free water. The quality and the concentration of the DNA was examined spectrophotometrically . The isolated DNA from the individual samples (n = 49) were individually sequenced, but analysed on nest level (n = 23).Total community DNA was isolated from great tit faecal samples (Following the recommendations of 1) followed by initial quality filtering using Trimmomatic v0.38 by cutting reads with an average quality per base below 15 using a 4-base sliding window and discarding reads with a minimum length of 200 bp , and to normalise for a possible nestbox effect and difference in the number of individuals sampled per nest (n = 23). Microbiota alpha diversity (Chao1 richness estimator and Shannon diversity estimator) measures were calculated using the phyloseq package (R = −0.43) at a time. For all models, significant effects were determined by ANOVA (type I sum of squares). The significance of the fragmentation effects for the different tree richness or tree composition levels was assessed using the addSE packages  . Due to per nest , nestlin package  and show package . Models packages . The effphyloseq, taking Jaccard (presence/absence), Bray–Curtis , unweighted UniFrac (phylogenetic distance) and weighted UniFrac  dissimilarities into account. The effect of either tree species composition or tree species richness and forest fragmentation (fragment area or edge density) on the microbial community composition was assessed via permutational multivariate analysis of variance (PERMANOVA) using the adonis function from the vegan package  are driving the difference in microbial community composition in monocultures with varying edge densities by taking into account the interaction between the tree species richness and the edge density and (2) are linked to host characteristics by considering either average fledgling SMI per nestbox or nestbox fledging success. Significant differences were obtained using a Wald test followed by a Benjamini–Hochberg multiple hypothesis correction. For all tests, a Bird captures and handling were carried out under licence and guidelines of the Belgian Ringing Scheme and the Flemish authorities . All trapping and sampling protocols of great tits were approved and permitted by the Ethical Committee VIB (the Flanders Institute for Biotechnology) Ghent site (EC2015-023) with permissions of all site owners.Staphylococcus, Bacillus, Lactobacillus, and Carnobacterium belonging to the Class Bacilli, together with genus Burkholderia-Paraburkholderia belonging to the Class Betaproteobacteria were most prevalent in the great tit microbiome , Proteobacteria (27.58 ± 8.44%), and Actinobacteria (17.90 ± 12.38%). Bacteroidetes (1.16 ± 1.09%), Tenericutes (1.49 ± 4.33%), and other phyla (0.91 ± 1.28%) showed a lower abundance , 2. Whencrobiome .F-value = 4.157; DF = 2; p-value = 0.034) and tree species composition with edge density on the Shannon diversity  was observed . When taking the interaction between tree species and edge density into account, a significant influence was observed on the alpha diversity, indicating an edge density effect on microbial richness with a direction depending on the particular tree species richness or composition . A signiobserved . More spobserved .p = 0.041) to 37.4%  of the variation observed between the nests. The combined effects of tree species richness and edge density could explain 23.2%  to 19.7%  of the variation .Permutational multivariate analysis of variance analysis on the beta diversity showed that the interaction between tree species composition and edge density significantly affected the UniFrac dissimilarity indexes, and accounted for 36.2%  differential abundance in monocultures when the edge density varies (Lactococcus (Class: Bacilli) was show to be most prevalent in monoculture plots characterised by a small edge density. Pseudarthrobacter, Arthrobacter (Class: Actinobacteria), and Erysipelatoclostridium (Class: Erysipelotrichia) showed a higher abundance in monoculture plots characterised by a high density of edges , Serratia (Class: Gammaproteobacteria), Sphingomonas (Class: Alphaproteobacteria), Carnobacterium, Jeotgalicoccus (Class: Bacilli), and Tyzzerella 3 (Class: Clostridia), with fold changes (log2) that vary between −2.12 and −30.00  can be pathogenic in birds, these most common genera contain a multitude of beneficial bacteria , nor the beta diversity . However, previous studies have shown that Julian date can be a confounding factor influencing the microbiome in passerines  of -30, indicating that this genus is strongly associated with a reduction in fledging success. Tyzzerella 3 has previously been linked to diabetes  can be found below: The animal study was reviewed and approved by Ethical Committee VIB (the Flanders Institute for Biotechnology) Ghent site (EC2015-023).AM, LL, KV, DB, FP, DD, and EV conceived the study and participated in its design and coordination. RB and DD performed material preparation and sample collection. SVP, EG, and EV performed the sample analysis. EG, EV, and LH performed the statistical analysis. EV wrote the first draft of the manuscript. All authors commented on previous versions of the manuscript, and read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.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.


Fifty-six patients were operated and 54 (89%) had a curative resection. Thirty-three patients (54%) had pT0-2, and 18 (30%) had pN0 tumours, with two patients achieving a complete response. As of 20 December 2020, 39 patients died, 32 due to GC, one within 30 days of surgery, two due to intestinal obstruction at 5 and 3 months after surgery and four due to other causes. Five-year survival rate was 38%. We conclude that perioperative chemotherapy is feasible in public hospitals in Chile and should be offered to patients with locally advanced GC.Gastric cancer (GC) is a leading cause of cancer death in Chile. Although recommended in international guidelines since 2006, perioperative chemotherapy was not available to patients in the public health system in Chile until 2016. We conducted an observational study to assess the feasibility of this strategy in public hospitals in Chile . Patients with locally advanced, operable GC were offered to receive preoperative chemotherapy with Epirubicin + Cisplatin + Capecitabine (ECX) for three cycles followed by curative surgery. Staging included abdominal CT scan and laparoscopy if peritoneal carcinomatosis was suspected. Postoperative ECX for three cycles was recommended. Between August 2010 and March 2013, 110 patients were screened and 61 enrolled. Median age was 62 years (23–76 years) and most patients had good performance status at baseline (Eastern Cooperative Oncology Group performance status score (ECOG) 0: 42, ECOG 1: 19). Tumour site was proximal in 32 (52%) and medial and distal in 29 (48%) patients. All but four patients ( Worldwide, gastric cancer (GC) is one of the leading cancers in incidence, being fourth and seventh in males and females, respectively. GC represents the third  and fifth  most common cause of cancer deaths globally . DespiteAt the time of diagnosis, the majority of patients present locally advanced or metastatic disease, while early lesions are diagnosed in less than 10% of cases . SurgeryTo improve cure rates, different multimodality treatments have been studied, establishing adjuvant chemoradiation , adjuvanIn 2006, the Chilean national cancer action plan included GC in the list of prioritised diseases within the Explicit Guarantees System . HoweverAdditional barriers for preoperative treatments in GC were: (i) lack of reimbursement for chemotherapy costs, (ii) patients were often presented at the multidisciplinary tumour board after surgery had already been performed and (iii) surgeons were concerned about progressive disease while on chemotherapy or an increase in surgical morbidity after chemotherapy, made it nearly impossible to offer a preoperative treatment strategy to patients in the public health system in Chile.To provide data on the feasibility of perioperative chemotherapy in Chile, we conducted a prospective, observational study to evaluate the efficacy and safety of perioperative chemotherapy with the ECX regimen (Epirubicin + Cisplatin + Capecitabine) at public hospitals  – GOCCHI-2009-01 – NCT0163320).Patients referred for neoadjuvant chemotherapy were screened. Patients older than age 18 diagnosed with histologically proven, locally advanced gastric carcinoma (cT3 or 4 and/or cN+), with no evidence of distant metastasis (cM0), and with an operable tumour, were eligible for this study. The 6th Edition of the American Joint Committee on Cancer (AJCC) Cancer Staging Manual was used . The resThe protocol was approved by the Ethics Committee of the Northern Metropolitan Health Service and all patients signed the approved informed consent form prior to treatment. All patients who signed the consent form and met all eligibility criteria were enrolled in the study. The chemotherapy drugs Epirubicin and Cisplatin for the preoperative treatment were donated by LKM, and the Instituto Nacional del Cáncer (Incancer) provided Capecitabine.2 IV Day 1 + Cisplatin (LKM) 60 mg/m2 IV Day 1 + Capecitabine (Xeloda®) 1,250 mg/m2 orally (PO) Day 1–21 (ECX) every 21 days for three cycles. Treatment delays and dose reductions were standardised. Curative surgery with total or subtotal gastrectomy and D2 lymphadenectomy was planned 3–6 weeks after the last dose of chemotherapy. Postoperative chemotherapy with the same regimen was recommended, but the medication could not be provided free of charge to the patients. After completion of perioperative treatments, patients were followed at months 6 and 12 after surgery and then yearly, with clinical evaluation. Imaging studies or endoscopy was performed on suspicion of progression.Patients received preoperative chemotherapy with Epirubicin (LKM) 50 mg/mThe primary endpoint was downstaging defined as the proportion of patients having T<3 tumours or N0 disease after three cycles of preoperative chemotherapy. Initially, downstaging was planned to be done by imaging, comparing baseline CT scan with a second CT scan done before surgery, but given the limitation of staging by CT imaging, final analysis for T and N stage was done based on pathological staging. Toxicity was a secondary endpoint and was assessed at day 1 of every chemotherapy cycle and prior to surgery using the CTCAE v3.0 manual . ResponsSource data were documented in the clinical charts at the Incancer using specially designed templates to ensure the completeness of data documentation for patient eligibility and clinical visits. Other source documents like signed informed consent forms, radiology and pathology reports were included in the patient files. Anonymised data were entered on electronic Case Report Forms (eCRFs) using a Good Clinical Practice (GCP), 21 CFR Part 11 compliant platform OpenClinica©. The eCRFs were designed by the study researchers and organised into clinical events, each with a single repetition . The use of eCRFs prevents data entry errors, allows one to easily keep track of form changes, helps solve data entry discrepancies and improves overall clinical team coordination. After data entry and review, reports were extracted from OpenClinica© as spreadsheets, and Matlab  was usedp-value < 0.05). For univariate survival analysis, the Cox proportional hazards model was applied. Statistical analysis was performed using Stata v15.0.Kaplan–Meier curves for overall survival were generated. To assess statistical differences between variables, the log-rank test was used , and the median time from staging CT to enrolment was 31 days (range: 5–83 days).In approximately half of the patients, the tumour involved the proximal portion of the stomach and/or the gastro-oesophageal union, while the other half of patients had non-cardia GCs. One third of patients had the presence of signet cells reported on the diagnostic biopsy. Almost two thirds of the patients had an intestinal, and only 15% a diffuse histological type. Given that many pathologists used the classification of the WHO, papillary, tubular and mucinous adenocarcinomas were classified as intestinal type, whereas signet-ring cell carcinomas were classified as diffuse type. Mixed carcinomas, carcinomas and adenocarcinomas not otherwise specified (NOS) were classified as indeterminate type.HER-2 status by immunohistochemistry could be obtained for 23 patients at baseline. In four cases (17%), overexpression of HER2 receptors was observed, one case (4%) was equivocal and 18 cases (78%) were negative.n = 57, 93%) completed three cycles of preoperative chemotherapy: one patient refused chemotherapy after the first cycle, one patient presented a cerebrovascular accident after the second cycle and two patients progressed. One patient had progressive disease after three cycles of chemotherapy and was not submitted to surgery.Most patients  were referred from the Hospital San Borja Arriarán, other referring centres were the Hospital San José (nine patients), the Incancer (eight patients) and the Hospital Rancagua (one patient). Chemotherapy was administered for all patients at the Incancer, while surgery was performed at the referring centre. All but four patients  had a curative resection, with R0 resection being confirmed in 51 (91% of patients operated). The median time from enrolment to surgery was 16 weeks (range: 11–30 weeks). Total gastrectomy was the most frequently performed surgery and the minority of patients had a splenectomy and/or pancreatectomy. Of 54 patients with curative resection, two had less than 15 lymph nodes (LNs) resected (4%), and the majority of patients (78%) had more than 25 LNs resected. One patient died within 30 days of surgery because of septic shock .Fifty-six patients (92%) were operated on. Details of the type of surgery performed are shown in After surgical resection, 27 patients started postoperative chemotherapy (50% of resected patients). Twenty-two patients (81% of the patients who started adjuvant chemotherapy) completed the planned three cycles, and five patients stopped early, three after one cycle and two after two cycles.Toxicity for pre and postoperative chemotherapy is shown in Clinical staging and pathological staging details are shown in Pathological TNM staging revealed tumours not penetrating the serosa or beyond (ypT0–2) in 33 patients (61% of resected patients) and 18 patients (33% of resected patients) who had no LNs involved after neoadjuvant chemotherapy (ypN0). A complete pathological response (ypT0N0) was found in two patients.As of 20 December 2020, 22 patients were still alive, 32 patients died due to the disease and 7 patients died due to other causes.Twenty-three patients completed the planned 5 years of follow-up. One of them was alive with recurrence, and 22 had no evidence of disease recurrence at 5 years. Median survival was 2.7 years with 3- and 5-year survival rates of 46% and 38%, respectively . Univariet al 7] and 34This finding suggests that, if multimodality treatment is planned, giving systemic therapy first will give the benefit of more than one treatment strategy to more patients compared to primary surgery. Our cohort included a slightly higher proportion of patients with involvement of the cardias or upper third of the stomach, compared to the series of the main referring centre  with suret al [In our study, of 110 patients screened, 49 (45%) did not meet the inclusion criteria, one third because of a calculated creatinine clearance of less than 60 mL/minute. Kuhnle et al  recentlyOur study has several limitations. We could not determine the clinical response after three cycles of preoperative chemotherapy, given the difficulty for accurate TNM staging by CT scan, particularly because CT scans were performed in different centres without standardised protocols. But even with such protocols, the accuracy of radiologic T and N staging after neoadjuvant chemotherapy is low . Only haHER-2 testing was done only in a subset of 23 patients, and no complementary FISH assay could be performed for two cases with equivocal findings. Interestingly, three of four patients with HER2 immunohistochemistry (IHC) 3+ staining are still alive, thus raising the question about the prognostic impact of HER2 positivity in GC treated with perioperative chemotherapy, as these patients were excluded in the recent report by Martinez Lago .This study provides important information to the public health authorities, supporting the decision to mandate financial coverage for perioperative chemotherapy by the public and private health insurance since 2016. Additionally, it provides reassuring data for the surgical field in Chile, leading to increasing support by surgeons for this treatment strategy.Since 2020, the FLOT regimen has been available for our patients in the public sector, and we are looking forward to offering the survival benefit seen in the FLOT4 study to our patients.The PRECISO trial collected quality of life data , blood sOur study confirms with real world data, that perioperative chemotherapy is feasible in public hospitals in Chile, and should be offered as an alternative to primary surgery for patients with locally advanced GC. Observational studies like PRECISO have the potential to provide data to support the introduction of new cost-effective therapies in the standard of care in countries with health systems with limited resources.www.clinicaltrials.gov.NCT0163320 at The authors have no conflicts of interest to declare.MC acknowledges partial financial support from the National Agency for Research and Development (ANID) projects ICN09_015, PIA/ACT192015; Corporación de Fomento de la Producción  and Deutscher Akademischer Austauschdienst .AHC acknowledges Comisión Nacional de Investigación Científica y Tecnológica (CONICYT) - Fondo de Financiamiento de Centros de Investigación en Áreas Prioritarias (FONDAP) 1513001 and Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT) 1191928 from the National Agency for Research and Development (ANID), Government of Chile.


The aim of this study was to investigate the atomic composition and the proteome of the salivary proteins adsorbed on the surface of orthodontic metallic bracket. For this, the atomic composition of orthodontic metallic brackets was analyzed with X-ray Photoelectron Spectroscopy (XPS). The acquired bracket pellicle was characterized after brackets were immersed in human whole saliva supernatant for 2 hours at 37°C. Hydroxyapatite (HA) discs were used as a control. Acquired pellicle was harvested from the HA discs (n = 12) and from the metallic brackets (n = 12). Proteomics based on mass spectrometry technology was used for salivary protein identification and characterization. Results showed that most of the proteins adsorbed on the surface of orthodontic metallic brackets and on the HA discs were identified specifically to each group, indicating a small overlapping between the salivary proteins on each study group. A total of 311 proteins present on the HA discs were unique to this group while 253 proteins were unique to metallic brackets, and only 45 proteins were common to the two groups. Even though most proteins were unique to each study group, proteins related to antimicrobial activity, lubrication, and remineralization were present in both groups. These findings demonstrate that the salivary proteins adsorbed on the bracket surface are dependent on the material molecular composition. A constant concern in the orthodontic clinic is the high prevalence of active caries lesions in patients undergoing treatment. In fact, it has been noted that 30 to 70% of patients experience active caries lesions during orthodontic treatment , 2. AlthSeveral studies have looked at the factors that guide bacterial colonization on tooth surface –13. AmonPrevious studies indicated that the level of bacterial growth on the surface of the orthodontic brackets varies according to the material of the bracket -–18. In fSalivary pellicles are not limited to the surface of the tooth, they are also present on the oral mucosa , on prosThe mechanisms by which proteins adsorb to surfaces is heavily determined by physical-chemical properties of the surface . It is eThe purposes of this study were to investigate the proteome composition of the acquired protein pellicle formed on the surface of metallic orthodontic brackets, and to compare the bracket pellicle against the pellicle formed on enamel-like surfaces (HA).An X-ray Photoelectron Spectrometer (Kratos Axis Ultra Spectrometer) was used to atomically analyze the composition of orthodontic metallic brackets  and HA discs . Three brackets and three discs were cleaned by sonication, in 800 μL distilled water, for 5 min, and incubated for 2 h at 37°C in 800 μL MiliQ water. Subsequently, XPS atomic analysis was performed on the brackets and HA discs. The XPS equipment was calibrated with the same parameters for all measurements . The ins2). Subjects refrained from eating or drinking two hours before saliva collection. After collection, all samples were centrifuged at 14,000 x g for 20 min at 4°C. Following the centrifugation, whole saliva supernatant (WSS) was pooled for the next experiment. Protein concentration was measured by the bicinchoninic acid assay .Ethics approval was obtained from the Ethics Committee on Human Research at The University of Western Ontario (protocol number 16181E). All participants signed an informed consent. Clinical evaluation was performed to assure that all participants were free of tooth decay, periodontal disease or any other condition that could affect the salivary composition. A total of 10 mL of whole saliva from three healthy subjects , aged from 30–40 years old, was collected in the morning, between 9:00 AM and 10:00 AM, under masticatory stimulation using parafilm  and four HA discs  were incubated in 800 μL of distilled water for 2 h at 37˚C with gentle agitation (in 24 well culture plate). Subsequently, the same brackets and HA discs were incubated for 2 h at 37˚C in pooled WSS containing the equivalent of 800 μg of total protein (200 μg of protein/bracket or disc). Immediately after incubation, the HA discs and brackets were washed using distilled water to remove any weak binding salivary protein. Experiments were done in triplicate.Pellicle formed on the surface of brackets and HA discs was harvested by sonication in 300 μL of solution containing 80% acetonitrile, 19.9% water and 0.1% trifluoroacetic acid (TFA) for 5 min. This procedure was repeated three times. The eluted pellicle proteins from the three replicates from the same group  were pooled, respectively, and concentrated by a rotary evaporator. Samples were re-suspended in distilled water to 900 μL, and the total protein concentration was assessed by the micro bicinchoninic acid assay .Aliquots of 15 μg of harvested pellicle protein from each group were prepared. The dried samples were re-suspended in 50 μL of 4 M urea, 10 mM DTT and 50 mM ammonium bicarbonate at pH 7.8 and incubated for 1 hour at room temperature. Afterwards, 150 μL of 50 mM ammonium bicarbonate was added to all samples, followed by 3% (w/w) trypsin . Thereafter, samples were incubated for 16 h at 37°C. Finally, samples were dried in a rotary evaporator, de-salted by C-18 ZipTip® Pipette Tips , and subjected to mass spectrometry.m/z values 390–2000 MS/MS. Each survey scan (MS) was followed by automated sequential selection of seven peptides for CID, with dynamic exclusion of the previously selected ions.After trypsinization, samples were re-suspended in 97.5% distilled water/2.4% ACN/0.1% formic acid and then subjected to reversed-phase nano-liquid chromatography electrospray ionization tandem mass spectrometry (nLC-ESI-MS/MS), using a LTQ-Velos  mass spectrometer. Liquid chromatography separation was achieved using a C18 column of capillary fused silica , linked to the mass spectrometer through electrospray ionization. Peptides were eluted from the nanoflow RP-HPLC over a 65-min period, with linear gradient ranging from 5 to 55% of solvent B , at a flow rate of 300 nL/min, with a maximum pressure of 280 bar. The electrospray voltage was 1.8 kV and the temperature of the ion-transfer capillary was 300°C. The survey scan was set in the range of http://ca.expasy.org) using Proteome Discoverer 1.3 software and SEQUEST algorithm. Parameter Xcorr was used. Search results were filtered for a False Discovery Rate of 1%, employing a decoy search strategy utilizing a reverse database. Each sample was analyzed four consecutive times by the mass spectrometer. For positive identification, the same protein had to be identified in at least three runs.The acquired MS/MS spectra generated were searched against Human protein database , silicon (Si), chromium (Cr), copper (Cu), sodium (Na), boron (B), nitrogen (N), sulfur (S), niobium (Nb), and palladium (Pd) . The widA total of 356 proteins were identified in the pellicle harvested from the HA disc control group (HA), and 298 proteins from the metallic orthodontic brackets (MB) . Forty-fBased on the isoelectric point (pI) of the identified proteins, the metallic bracket pellicle had 67 proteins with pI below 6, 106 proteins had pI between 6–8, and 125 proteins had pI above 8. For the HA discs, 66 proteins had pI below 6, 128 had pI between 6–8, and 162 had pI above 8 .According to biological functions , proteinCaries lesions around and under orthodontic brackets is a common occurrence during orthodontic treatment which may cause delays in the treatment, increased costs, and irreversible loss of tooth tissue . The devPrevious studies have shown that the overall net charge of orthodontic brackets is usually neutral ; howeverInterestingly, most proteins were identified specifically to each group, suggesting specificity of the protein’s adsorption in relation to the surface of the bracket. As a result, biofilm formation  and bactStreptococccus mutans [S. mutans when compared to metallic brackets [In fact, amylase was uniquely identified in the pellicle formed on the HA discs. A previous study suggested that salivary alpha-amylase binds very selectively and with high affinity to several streptococcal species . This iss mutans . Furthers mutans . Nonethebrackets , an obseAggregatibacter actinomycetemcomitans has been suggested for cystatin SA [Other proteins with high affinity for HA surfaces are cystatins . While ntatin SA .Candida albicans [On the other hand, histatin 1 was identified on both metallic brackets and HA discs. Histatin 1 possesses strong antifungal properties against albicans . In addialbicans , 38 and albicans  have beeSurprisingly, although statherin is known for its strong affinity for the tooth surface, this protein was identified in our study in the pellicle formed on metallic brackets. Statherin shows critical function in tooth remineralization as it is the only salivary protein to inhibit both primary (spontaneous) and secondary  calcium phosphate precipitation ; thus, mDifferent mucins were found in both groups. Mucins are proteins with high carbohydrate content. Overall, mucins play a role in lubrication, non-immune protection and they can modulate the adhesion of microorganisms to the oral tissue surfaces . MUC5B aBesides confirming the expected differences in the molecular composition of metallic orthodontic brackets and HA discs, this study reaffirmed the selectivity in the binding of salivary proteins to surfaces in the oral cavity. Moreover, this first characterization of the pellicle formed on the surface of metallic brackets, suggests that the metallic surface attracts important calcium-binding proteins, such as histatin 1 and statherin, which can assist in the remineralization process, while lacking proteins, such as amylase, with higher potential to bind cariogenic pathogens. Further investigation is needed to verify the effect of the characterized bracket pellicle on bacterial adhesion. Our results support the idea of surface modulation as an important path to favor the formation of bracket pellicles enriched with proteins with selected biological functions to prevent dental caries.Selective adsorption of proteins is observed in many different surface materials. Similarly, the adsorption of salivary proteins to the surface of orthodontic metallic brackets happens in a selective fashion. Interestingly, proteins with recognized high cariogenic characteristics were not identified on the metallic bracket pellicle, suggesting a protein pellicle with lower cariogenic activity. Further improvement in oral health and reduction of caries activity during orthodontic treatment may be explored via additional modulation of the acquired bracket pellicle by bracket surface modifications.S1 Table(DOCX)Click here for additional data file.