The more times people fail the more likely they are to give up, however little is known about the neural mechanisms underlying this impact of repeated failure on decision making. Here we have used a visual shape discrimination task with computer-controlled feedback combined with functional magnetic resonance imaging (fMRI) to investigate the neural circuits involved. The behavioral task confirmed that the more times subjects experienced failure the more likely they were to give up, with three successive failures being the key threshold and the majority of subjects reaching the point where they decided to quit and try a new stimulus set after three or four failures. The fMRI analysis revealed activity changes in frontal, parietal, temporal, limbic and striatal regions, especially anterior cingulate cortex (ACC), posterior cingulate cortex (PCC) and temporal parietal junction (TPJ) associated with the number of previous failures experienced. Furthermore, their parameter estimates were predictive of subjects’ quitting rate. Thus, subjects reach the point where they decide to quit after three/four failures and this is associated with differential changes in brain regions involved in error monitoring and reward which regulate both failure detection and changes in decision-making strategy. Reinforcement learning (RL) is a process whereby the estimation of state/action values is improved through trial and error so that behavior becomes more advantageous23458To behave adaptively, animals and humans need the capacity for reinforcement learning to internally monitor responses and to evaluate external reinforcement or feedback111314151618192042122The “three strikes” phenomenon is well known and has often been reported in research related to decision making or sports psychology24Trial-by-trial analysis revealed that whether subjects decided to continue or quit discrimination of a specific stimulus set was influenced by the number of times they received negative feedback. We used a logistic regression model to predict whether subjects continued or gave up after receiving different numbers of negative feedbacks. This simplified model used one factor (the number of negative feedbacks) and could be expressed asR2 = 0.286, Nagelkerke R2 = 0.451, accuracy of prediction = 84.2%. Based on the model, we drew a ROC curve with an area under the curve of 0.8 < S = 0.869 < 0.9, suggesting that the model diagnosis was good . Using Hosmer-Lemeshow analysis to test goodness of fit we found that there was no difference between the prediction result and initial data, which also indicated that the model was good . These results showed that whether to continue or to quit was indeed influenced by the number of times subjects received negative feedback.with the following functions: Logit(P) = Am*F, where F is the number of negative feedbacks, Am is the regression coefficient of negative feedback. The indices of the model were: Cox & Snell th, 5th, 6th or 7th negative feedback was received. With respect to quitting rate, a repeated-measures ANOVA revealed a main effect of the number of negative feedbacks, F3,57), other than between the first and second time (p = 0.170). These results showed that subjects reached the point where they started to change from continuing to quitting after the third time of receiving negative feedback. To further explore the relationship between the number of times negative feedback was received and quitting rate (percentage of quitting the item), we conducted a linear regression (y = ax + b) and a quadratic polynomial curve (y = ax2 + bx + c) separately. The linear regression function was y = 25.32x − 36.16, R2 = 0.74  the decision to quit occurred between 1 and 4 failures, with most subjects doing so after 3 or 4 failures. Only 7.04% of quitting decisions occurred after 5 failures, 1.52% after 6 and 0.11% after 7 see . In the 0.74 see . The fitvals see . To illuAn analysis of confidence levels after each failure showed that they decreased following each successive failure see . ImportaTo investigate which brain regions showed activity changes associated with negative feedback in the task, we first performed a whole-brain analyses on fMRI data collected during the feedback stage. Since there were too few trials where subjects had more than 4 failures to perform an effective analysis of fMRI data we only analyzed neural changes following 1, 2, 3 and 4+ failures. This analysis revealed that a number of middle and superior frontal, temporal and occipital cortical regions and the ventral striatum showed increased activity whereas medial and dorsolateral frontal regions showed decreased activity. These neural activity changes were significantly associated with the number of times negative feedback was received see . A numbeTo explore how activity changes in the different brain regions related to the number of times negative feedback was received and quitting rate, we used the same quadratic polynomial function as for analysis of the behavioral data. Brain activations were extracted from 8 mm diameter spheres in the following ROIs: MPFC , bilateral dACC , left insula , right DLPFC , left and right VStr , left PCC , right precuneus , right amygdala , left middle temporal gyrus  and right TPJ  .F3,76), i.e. they showed the same trend as the behavioral results. Post-hoc, Bonferroni-corrected analysis results showed that the parameter estimate for the first negative feedback was significantly different from the second (p = 0.048), the third (p = 0.01) and fourth (p = 0.005) respectively in dACC . For PCC , there were no significant differences between the first, second and third feedbacks (p > 0.05) but there was for the fourth one (p = 0.037). For the TPJ  there were no differences between the first two times of negative feedback (p = 0.119) but there were between the first and the third and fourth times .Repeated-measures ANOVA was used to test whether the parameter estimates of each target region were significantly different for each of the four frequencies of negative feedback . The results showed there were main effects of negative feedback in dACC, PCC and TPJ . Additionally, anatomical images (256 × 256 × 176) with 1 × 1 × 1 mm3 resolution were obtained by a T1-weighted three-dimensional magnetization prepared rapid gradient echo (MPRAGE) sequence .During each fMRI scan an Echo-Planar imaging (EPI) sequence was used for data collection, and T2*-weighted images were recorded per run using a Siemens TRIO 3.0T MRI scanner  were modeled to create the design matrix. The regressors were then convolved with the Canonical Hemodynamic Response Function and the six realignment parameters for each subject were also included as confounding factors.Statistical analyses were performed using the general linear model in SPM8p < 0.05 FWE corrected, with clusters of ≥10 contiguous voxels.Our results primarily relate to feedback processing We investigated brain regions whose activation was associated with the process of negative feedback using one-sample t-tests. Moreover, we also investigated the contrast between positive (correct and get points) and negative feedback (f1 + f2 + f3 + f4). In addition, the contrast between the decision to stay or shift after negative feedback during both the negative feedback and choice stages was conducted. The image threshold for fMRI data significance was set to, We used the MarsBar toolbox618p < 0.001, small volume corrected, with a minimum cluster size of 10 voxels.Based on behavioral results, we could infer that subjects tended to continue with the same stimulus set after up to 2 failures (f1 + f2) but would quit after ≥3 failures (f3 + f4). Functional connectivity strengths were then analyzed to investigate changes between regions associated with behavioral choice (i.e. continue or quit). We measured functional connectivity using gPPI analysis5558How to cite this article: Zhao, W. et al. Neural circuitry involved in quitting after repeated failures: role of the cingulate and temporal parietal junction. Sci. Rep.6, 24713; doi: 10.1038/srep24713 (2016).


Notoriously, the island of Mauritius has one of the highest prevalence of diabetes in the world. Management of the disease is very important and family meals are undoubtedly beneficial to patients as they promote the development of healthy eating behaviours and food choices. This study has aimed to probe into potential relationship(s) between family meal frequency and individual dietary intake among diabetic patients and to establish whether family cohesion may be a plausible mediator of this relationship.χ2) tests, independent sample t-tests and one-way ANOVA were used to determine statistical significance. Pearson correlation was used to examine associations between family meal frequency, individual dietary intake and family cohesion. Hierarchical linear regression models were performed for the mediation analysis.A cross-sectional survey was carried out with a random sample of 384 diabetic patients. The Family Adaptability and Cohesion Evaluation Scale III was used to obtain information on two general aspects of family functioning, that is, cohesiveness and adaptability. Chi-squared (P < 0.05).Family meal frequency  was observed to be positively associated with intake of fish, raw vegetables, dried and fresh fruits, low-fat milk, cheese, yogurt, nuts and light butter and negatively associated with intake of red meat, white rice, white bread, whole egg fried, chocolates, fried cakes, burgers, chips, and fried noodles/rice. Average mediation (52.6 %) was indicated by family cohesion for the association between family meal frequency and individual dietary intake among diabetic patients. Sobel’s test further confirmed the trend towards complete mediation (z = 15.4; A strong relationship between family meal frequency and individual dietary intake among diabetic patients was recorded. The present study is one of the few studies that have examined family cohesion as a mediator of the relationship and to our best knowledge is the first work to demonstrate a trend towards complete mediation. Results obtained can be used by health professionals to devise strategies for increasing knowledge and awareness of both diabetic patients and their respective families to curd down this public health burden. The World Health Organisation (WHO) defines diabetes as a chronic metabolic disease that occurs either when the pancreas does not produce enough insulin or when the body cannot effectively use the insulin it produces . DiabeteAbout 382 million people, in 2013, have diabetes in the world and it is expected to rise up to 592 million by 2035 if serious attention is not drawn towards the problem . MauritiPharmacotherapy is normally used by most diabetic patients as the primary means of achieving optimal glycaemic control. However, most drugs are usually associated with weight gain. Therefore, medical nutrition therapy (MNT) and lifestyle intervention remains the cornerstone for the management of diabetes, especially those with NIDDM . Along wPrevious studies have focused mostly on the influence of family meals on the consumption of fruits and vegetables in adolescents. Moreover very few studies have been conducted on the relationship between family meal frequency and individual dietary intake and therefore there is no established theory to explain the association among diabetic patients in Mauritius. Family cohesion, defined as the emotional bonding that family members have towards one another, may have significant implications for the adoption of healthful eating habits . It can A self-administered cross-sectional survey was conducted to meet the objectives of the study since it is a quantitative data collection approach targeted to obtain information from a representative sample of the population. For a population of about 290,000 diabetic patients in Mauritius, the requested sample size, for ± precision with 95 % confidence interval (CI) and degree of variability, p equal to 0.5, was found to be 384. To make the sample representative of the population suffering from diabetes mellitus, participants were recruited randomly from health care centres all around the island from both rural and urban areas. Non-governmental organizations such as Mauritius Diabetes Association, T1diams, Diabetes Parent Support Group and ‘Groupement des Diabétiques’ were also targeted.The respondents selected for the study had to meet the following inclusion criteria: (1) Type I and Type II diabetic patients only were recruited; (2) patients above 12 years; (3) Participants who are mentally fit to answer questions; and (4) Participants should be of Mauritian origin and they should be able to understand English, French and Creole (vernacular) language. The exclusion criteria included patients suffering from other types of diabetes, except Type I and Type II. Diabetic patients suffering from extremely poor health conditions and having difficulties to understand and answer questions were also excluded.The questionnaire comprised of 20 main questions and divided into four distinct sections; personal data such as socio-demographic information; individual dietary intake; family meal frequency and family support and cohesion .2). The weight of participants, with their shoes removed, was recorded by the first author using a calibrated digital scale and the height was measured using a measuring tape.Section A consisted of questions based on personal and socio-demographic information such as age, sex, ethnic group, residential area, educational background, occupation, income group, marital status and number of persons in the family which are considered to be predictors of self care behaviour. Questions on types of diabetes, family history of diabetes and body mass index (BMI) were also being asked. BMI was calculated as weight (kg)/height squared  had breakfast, lunch and dinner together?” Responses were accepted as never, 1–2 times, 3–4 times and every day.http://www.eruralfamilies.org]. Family cohesion assesses degree of separation or connection of family members to the family. There are four levels of family cohesion ranging from extreme low cohesion to extreme high cohesion- disengaged, separated, connected, and enmeshed. There are four levels of adaptability - rigid, structured, flexible, and chaotic [Section D consisted of questions on family support and cohesion. The Family Adaptability and Cohesion Evaluation Scale III (FACES III) is a validated 20-item self-reporting scale which was used to obtain information on two general aspects of family functioning, that is, cohesiveness and adaptability [12  chaotic . Through chaotic . Therefo chaotic .Table 1CThe questionnaire was designed in English language and translated in Creole, a commonly spoken local language in Mauritius, or French for those who were not comfortable with English language. However, in cases where participants had language and communication issues, data was collected using face to face interviews where the questionnaire was filled out by the first author. Participation to the study was voluntary and formal consent was obtained from all the respondents. An informed consent form was provided at the beginning of each questionnaire where they were assured of the confidentiality of the data collected , 17.χ2) test, independent sample t-tests and one-way ANOVA analysis were performed to compare the various socio-demographic variables, family cohesion scores and eating behaviours of diabetic patients. Pearson correlation coefficients were used to examine the associations between family cohesion and family meal frequency. The Baron and Kenny approach as described by Welsh et al. [To demonstrate that the independent variable X  was associated with the outcome variable Y .To demonstrate that the independent variable X was associated with the mediator variable M (family cohesion).To demonstrate that the mediator variable M was associated with the outcome variable Y after controlling for X.The last step consisted of determining whether the effect of X on Y was reduced when M was included in a regression model, providing evidence that the effect of X on Y was partially mediated by M. Furthermore, if the path between X and Y disappeared completely when M was included, complete mediation by M was indicated see Fig. .Data was analysed using SPSS (version 16.0) software and Excel 2007. Chi squared . Similar regression models were fit regressing family cohesion scores on frequency of family meals as well as regressing average servings of targeted food or beverage items on family cohesion scores after controlling for frequency of family meals. Regression models regressing weekly servings of a targeted food category on both family meal frequency and family cohesion score were fit in order to provide evidence for mediation. When mediation was indicated, the percentage reduction for the association of family meals and individual dietary intake as mediated by family cohesion was calculated by subtracting the adjusted regression coefficient for family meal frequency from its unadjusted counterpart and then dividing by the unadjusted regression coefficient for frequency of family meals . For eacA total of 384 participants completed the questionnaire which was representative of the Mauritian population. Socio-demographic variables such as gender, age group, ethnic group, demographic area, marital status, educational background, occupation, income group, number of persons in family, type of diabetes and family history of diabetes are displayed in Table The BMI was calculated from the measured height and weight of the respondents and Fig. The eating behaviours and food consumption of the participants were assessed using different questions on vegetarianism, number of meals per day, skipping meals and knowledge of glycaemic index. The Food Frequency Questionnaire indicated the food intake of specific food items. It was found that 99.0 % of the subjects were non-vegetarian and only 1.0 % were vegetarian. 52.6 % consumed “3 meals” per day while the rest had “less than 3 meals”, “5-7 meals” and “more than 7 meals” with percentage occurrence of 9.4 %, 28.9 % and 9.1 % respectively . The majority of the respondents were from a disengaged family (i.e. extreme low cohesion) with an occurrence of 51.6 %. 26.6 % came from an enmeshed family, 14.1 % from a connected family and 7.8 % from a separated family see Fig. . The meaP <0.05) between BMI and the following socio-demographic variables  was recorded. However, no significance was recorded between BMI with respect to the following demographic variables; sex (P = 0.554), marital status (P = 0.913), income group (P = 0.119), number of persons in family (P = 0.104) and type of diabetes (P = 0.973).A strong statistically significant relationship (P = 0.000), occupation (P = 0.000), income group (P = 0.000) and type of diabetes (P = 0.001) with respect to number of meals per day was recorded. However no statistically significant relationship was noted for sex (P = 0.367), ethnic group (P = 0.054), demographic area (P = 0.060), marital status (P = 0.099), educational background (P = 0.126), number of persons in family (P = 0.347) and family history (P = 0.562) with respect to number of meals per day.A strong significant relationship between age group (P = 0.000) and educational background (P = 0.000) with respect to skipping meals was recorded. However, no statistically significant relationship was noted for sex (P = 0.612), ethnic group (P = 0.078), demographic area (P = 0.341), marital status (P = 0.893), occupation (P = 0.073), income group (P = 0.674), type of diabetes (P = 0.075) and family history (P = 0.088) with respect to skipping meals.A strong statistically significant relationship between age group (P = 0.001), occupation (P = 0.000), type of diabetes (P = 0.001) and family history (P = 0.008) with respect to glycaemic index was recorded. However no statistically significant relationship was found between sex (P = 0.454), ethnic group (P = 0.093), demographic area (P = 0.212), marital status (P = 0.562), educational background (P = 0.229), income group (P = 0.208), number of persons in family (P = 0.520) with respect to glycaemic index.A strong statistically significant relationship between age group (P = 0.001), beef (P = 0.000) and fish (P = 0.001) was recorded. Moreover, statistically strong relationship was found between demographic area and chicken (P = 0.000) and lamb (P = 0.000) consumption. Strong relationships were also found between income group and lamb (P = 0.001), brown bread (P = 0.001) and burger consumption (P = 0.001). Interestingly, the results also revealed strong statistically significant relationship between age group and burger (P = 0.000), chips (P = 0.000), pizza (P = 0.020), briani (P = 0.009) and fried noodles/rice consumption (P = 0.046). Strong relationships were equally found between educational background and burger (P = 0.000), chips (P = 0.000) and pizza consumption (P = 0.000). However there was no statistically significant relationship between the socio-demographic variables and the other remaining food items.A significantly strong relationship between ethnic group and chicken consumption (r = 0.850), “eat lunch together” (r = 0.895) and “eat dinner together” (r = 0.827) with respect to family cohesion score was recorded.Table r = −0.631), white rice (r = −0.610), whole egg fried (r = −0.644), gateaux-piments (r = −0.674), burger (r = −0.657), chips (r = −0.669), briani (r = −0.671) and fried noodles/rice (r = −0.677) consumption with respect to perceived family cohesion score was recorded. Moreover, strong positive association was found between egg without yolk (r = 0.659), dried fruits (r = 0.731), low-fat milk (r = 0.682), cheese (r = 0.611), yogurt (r = 0.671), light butter (r = 0.627) with respect to perceived family cohesion score. Interestingly, very strong correlations were found between fish (r = 0.839), raw vegetables (r = 0.804), fresh fruits (r = 0.807) and nuts (r = 0.877) consumption with respect to family cohesion scores.Strong negative correlation between lamb/mutton (r = 0.863) and nuts (r = 0.893) consumption with respect to family meal frequency. For those who ate lunch together, there was a strong negative association between lamb/mutton (r = −0.622), white rice (r = −0.658), white bread (r = −0.681), whole egg fried (r = −0.633), gateaux-piments (r = −0.691), burger (r = −0.666), chips (r = −0.676), briani (r = −0.681) and fried noodles/rice (r = −0.694) consumption with respect to family meal frequency. There was a statistically strong positive correlation between egg without yolk (r = 0.721), dried fruits (r = 0.709), low-fat milk (r = 0.652), yogurt (r = 0.681) and light butter (r = 0.658) consumption with respect to family meal frequency. It was also seen that there was a very strong positive association between fish (r = 0.873), raw vegetables (r = 0.875), fresh fruits (r = 0.863) and nuts (r = 0.919) consumption with respect to family meal frequency. For those diabetic participants who had dinner with their family, a strong negative correlation was found ranging from −0.612 to −0.795.For those who had breakfast together, there was a strong negative association ranging from −0.643 to – 0.716 with respect to family meal frequency. It was also noted that there was a strong positive correlation between raw vegetables ranging from 0656 to 0.788 with respect to family meal frequency. In addition, results equally demonstrated a very strong positive association between fish  and their standard error (SE) for the hierarchical linear regression models regressing average number of servings of the targeted food items  on family meal frequency in the three categories  are displayed in Table Regression coefficients (β) and their standard error (SE) for the hierarchical linear regression models regressing family cohesion scores on family meal frequency in the three categories  was carried out. The family cohesion score demonstrated strong positive associations when regressed on family meal frequency (p <0.005).Regression coefficients (β) and standard error (SE) for hierarchical linear regression models regressing average number of servings of the targeted food items on family cohesion scores was carried out. Significant positive associations for family cohesion scores and fish (β = 0.161), egg without yolk (β = 0.081), raw vegetables (β = 0.352), dried fruits (β = 0.117), fresh fruits (β = 0.340), low-fat milk (β = 0.189), cheese (β = 0.115), yogurt (β = 0.150), nuts (β = 0.211) and light butter (β = 0.090) were observed.Regression coefficients (β = −0.227), white rice (β = −0.207), whole egg fried (β = −0.226), burger (β = −0.174), chips (β = −0.176), briani (β = −0.180) and fried noodles/rice (β = −0.180).Significant negative associations were found for family cohesion scores and lamb/mutton (β) and standard error (SE) for hierarchical linear regression models regressing average number of servings of the targeted food items on family cohesion scores, after controlling for frequency of family meals. It can be deduced that significant positive associations continued to be observed for family cohesion scores and intake of fish, egg without yolk, raw vegetables, dried fruits, fresh fruits, low-fat milk, cheese, yogurt and nuts after controlling for frequency of family meals, that is, “eat breakfast together”, “eat lunch together” and “eat dinner together”.Table P = 0.000.These findings indicate a 52.6 % reduction for the association of family meal frequency and individual dietary intake of the targeted food items as mediated by family cohesion. The calculation was done by subtracting the adjusted regression coefficient for family meal frequency from its unadjusted counterpart and then dividing by the unadjusted regression coefficient for frequency of family meals. The percentage reduction obtained clearly indicated a more or less complete mediation (>50 %) for the association between family meal frequency and individual dietary intake as mediated by family cohesion. Sobel’s test, carried out using SPSS version 16.0, confirmed the trend towards complete mediation by family cohesion for the relationship between family meal frequency and individual dietary intake as indicated with z = 15.4 and The main aim of this study was to examine the relationship between family meal frequency and individual dietary intake among diabetic patients in Mauritius. In addition to the primary aim, it was also hypothesised that family cohesion may be a plausible mediator of this relationship. To our best of knowledge, no such studies have evaluated the association between family meal frequency, individual dietary intake and family cohesion among diabetic patients in Mauritius. It was hypothesised that family meal frequency would be positively associated with healthy food items intake  and negatively associated with less healthy food items intake . Moreover, an endeavour has been made to assess a series of socio-demographic variables including sex, age group, ethnic group, demographic area, marital status, educational background, occupation, income group, number of family members, type of diabetes and family history of diabetes in relation to glycaemic index, skipping meals, number of meals per day, intake of targeted food items and BMI.6 and negative associations with fast- food intake were also found by Burgess-Champoux et al. [This study has shown that 40.9 %, 28.1 % and 45.3 % of the diabetic patients had breakfast together, eat lunch and dinner together respectively. Mellin et al.  have foux et al. , which wx et al.  have demp < 0.05) of the efficacy of treatment in controlling diabetes. According to Geffken et al. [To our best of knowledge, no such studies have examined the association between family cohesion, family meal frequency and dietary intake among diabetic patients. Studies previously done on family cohesion are relatively few and none of them have really investigated on dietary consumption. Combrey et al.  have shon et al. , higher n et al.  found thHowever, only one study investigated the association between family cohesion and individual dietary intake. Welsh et al.  have shoWelsh et al.  have alsThis study has also taken into account the socio-demographic variables in relation to BMI, skipping meals, glycaemic index, number of meals per day and the average servings of the targeted food items. Our findings indicated that most of the diabetic patients actually consumed three meals per day (52.6 %) instead of the recommended number of meals, that is, five meals per day 28.9 %). It was also noted that 60.9 % of the participants did not skip meals and only 39.3 % actually did skip meals. Significant differences were found between age group and skipping meals as well as for education and skipping meals. A decline in breakfast consumption by mostly adolescents has been documented in the United Sates . The rea %. It waStatistically significant relationships were noted for age group, ethnic group, demographic area, education, occupation and family history in relation to BMI. O’Neil et al.  observedInterestingly, our results have shown that 97.4 % of the diabetic patients did not have any knowledge about glycaemic index and only 2.6 % were aware of the meaning of glycaemic index. Many studies have shown the importance of consuming a low- glycaemic index meal in order to control blood glucose levels and unfortunately in the present study most of the respondents were unaware of the situation. Yusof et al.  has founIncome was a determining variable in the consumption of certain food items. There were statistically significant relationships between income group and consumption of lamb/mutton, brown bread and burgers. Deshmukh-Taskar et al.  have fouThe specificity of the study is the use of a validated method, Family Adaptability and Cohesion Evaluation Scale III (FACES-III) by Baron and Kenny , to measEven though our findings precisely demonstrate a relationship between family meal frequency and individual dietary intake, future studies can formulate more effective evaluation tools that accurately measure the frequency of family meals and the quality of meals being served at home. Since not many studies have been conducted on these three parameters, several questions ultimately stem from the results of this study indicating a need for immediate and positive intervention within the field of family meal environment and connectedness.A decline in family meals across generations has been observed and this trend may not necessarily be due to the fact that family members do not enjoy eating together. Busy work schedules, children’s activities after school, television viewing and new methods of processing and distributing foods such as pre-packaged meals may all be contributing factors to the demise of family members sitting together for a meal. Previous research has investigated and demonstrated the numerous benefits of family meals including greater consumption of healthy foods, adolescent’s health status, glycaemic control and lower fasting total cholesterol. Our findings have shown that greater frequency of family meals was significantly associated with greater consumption of healthy foods such as fish, raw vegetables, dried fruits, fresh fruits, low-fat milk, cheese, yogurt, nuts and light butter, that is, better individual dietary intake. Thus it can be recommended that specific nutrition intervention programs encouraging the frequency of more family meals need to be targeted to the diabetic population as well as their respective families. Furthermore, the measurement of family cohesion as a plausible mediator of this relationship adds to the uniqueness of the study. We recorded a 52.6 % reduction for the association of family meal frequency and individual dietary intake as mediated by family cohesion which indicated a more or less complete mediation. Henceforth, it can be concluded that health professionals should take into consideration the connectedness of family members in order to devise innovative nutrition intervention strategies. Therapeutic and psychological interventions, involving family members, can be adopted for the management of diabetes. However, future research in this area is needed to assess other mediators such as parents’ rules at home, time spent in food preparation and availability of home-made meals.


IMAGE  included a retrospective cohort to describe real‐world treatment prior to approval of the immune checkpoint inhibitor ipilimumab. This retrospective cohort of patients, who started second‐line/subsequent treatment (index therapy) for advanced melanoma within 3 years before ipilimumab approval, was selected randomly by chart review. Collected data included treatment history, patient outcomes, and healthcare resource utilization. All patients had ≥1 year of follow‐up data. This analysis included 177 patients from Europe (69%) and North America (31%). The most common index therapies  were fotemustine (23%), dacarbazine (21%), temozolomide (14%), and platinum‐based chemotherapy (14%). Most patients (89%) discontinued index treatment during the study period; the most common reason was disease progression (59%). Among patients with tumor assessment , 2% had complete response, 5% had partial response, and 12% had stable disease on last tumor assessment. At 1‐year study follow‐up, median progression‐free survival was 2.6 months  and median overall survival was 8.8 months . During follow‐up, 95% of the patients had healthcare visits for advanced melanoma, 74% of whom were hospitalized or admitted to a hospice facility. These results provide insights into patient care with advanced melanoma in the era before ipilimumab and may serve as a benchmark for new agents in future real‐world studies.The therapeutic landscape for advanced melanoma has recently been transformed by several novel agents (immune checkpoint inhibitors and molecular‐targeted agents). The prospective, multi‐site, observational study  Melanoma poses a great clinical challenge BRAF V600 mutant population. Nivolumab The therapeutic landscape for advanced melanoma has recently been transformed by the approval of several novel agents (immune checkpoint inhibitors and molecular‐targeted agents) that are more effective than conventional therapies imumab: management of advanced melanoma in real practice; ClinicalTrials.gov Identifier: NCT01511913) study is a multi‐site, observational study evaluating real‐world treatment and patient outcomes for advanced melanoma, both prospectively and retrospectively. This study describes the results from the retrospective cohort, which was treated in the era before ipilimumab and may serve as a benchmark for new agents in future real‐world studies.The IMAGE , tumor response rate, and healthcare resource utilization among these patients.This study was conducted at sites in Europe  and North America (Canada and the United States). Data obtained from patient charts were entered by all sites into electronic case‐report forms, with monitoring for verification of the source data. Data entry was expected at a minimum frequency of every 3 months, and data were collected for each patient for ≥1 year from start of index therapy (defined as second‐line or later treatment initiated on entry into the study). Data were extracted on 15 September 2014.This study was conducted in accordance with the International Society for Pharmacoepidemiology Guidelines for Good Epidemiology Practices and applicable local regulatory requirements, and adhered to the guidelines for company‐sponsored, postauthorization, safety studies as outlined by the European Medicines Agency in the Guideline on Good Pharmacovigilance Practices (GVP)—Module VIII. The protocol was approved or acknowledged  by the Institutional Review Board or Ethics Committee at each participating site.Eligible patients had to have been previously treated for advanced disease, and study entry was defined as start of the index therapy within 3 years before the approval of ipilimumab. Index therapies therefore began between 25 March 2008 and 01 February 2012 .The retrospective cohort of patients was selected via chart review based on the following criteria: diagnosis of unresectable or metastatic melanoma, aged ≥18 years at the time of entry into the study, receipt of at least one prior therapy for unresectable or metastatic melanoma, initiation of second or subsequent therapy for unresectable or metastatic melanoma within the 3 years prior to the approval of ipilimumab, and a minimum of 1‐year follow‐up data available regardless of patient's survival status. First‐line therapy did not need to occur in the 3‐year period prior to ipilimumab approval or after the diagnosis of unresectable or metastatic melanoma. Although first‐line therapy could have occurred in the 3‐year window, a second‐line of therapy in that 3‐year period was required to qualify the patient.All retrospective cohort data were reported through the 1 year of study follow‐up. Patient demographics and baseline characteristics were reported using descriptive statistics, including mean and standard deviation (SD) for continuous variables, and count and percentage for categorical variables. Descriptive statistics were provided for index therapy, first‐observed prior melanoma therapy (defined as the first melanoma therapy prior to index therapy), and last‐observed prior melanoma therapy (defined as the last melanoma therapy prior to index therapy considering only patients with multiple prior therapies). Tumor response was based on the last (or only) tumor assessment record with nonmissing assessment date during the 1‐year study follow‐up period and was categorized as complete response, partial response, stable disease, progressive disease, or indeterminate based on response criteria applied during the study . Probabilities for PFS (defined as the time from the date that index therapy was initiated to the date of progression or death from any cause) and OS (defined as the time from the date that index therapy was initiated to the date of death from any cause) were estimated using the Kaplan–Meier product limit method. PFS and OS were reported as medians, with corresponding 2‐sided 95% confidence intervals (CIs) using the method of Brookmeyer and Crowley, and as means with SDs. Healthcare resource utilization, which included healthcare visits due to advanced melanoma and hospitalization and/or hospice facility visit, were reported using descriptive statistics.BRAF V600 mutation at baseline, 47% (17/36) were positive.A total of 177 patients , dacarbazine (21%), temozolomide (14%), and platinum‐based chemotherapy (14%)  patients had ≥1 tumor assessments during the follow‐up period, and last tumor response for these patients was complete response in 2% (3/153) and partial response in 5% (8/153) of the patients Table . A totalAlmost all patients 95%; 168/177) had a healthcare visit due to advanced melanoma during the 1‐year study follow‐up period  for advanced melanoma within 3 years before approval of ipilimumab were selected randomly by chart review.n = 750; the MELODY study), the most commonly used systemic treatments across all lines and outside the clinical trial environment were dacarbazine (51%), fotemustine (42%), and temozolomide (11%) α, and IL‐2 were also used in 14–22% of patients across all lines The findings in this study showed that a wide range of advanced melanoma therapies were used in the era before ipilimumab. The most common index therapies were fotemustine (23%), dacarbazine (21%), temozolomide (14%), and platinum‐based chemotherapy (14%), administered alone or in combination. The most common single‐agent index therapies were dacarbazine (19%) and fotemustine 18%). The treatment patterns in this study were generally consistent with those described in other real‐world studies conducted prior to the use of immune checkpoint inhibitors and molecular‐targeted agents. For example, in a larger European‐only study  and median OS was 8.8 months  at 1 year. These results were consistent with those reported historically in the melanoma literature Results from retrospective cohort analysis of the IMAGE study provide insights into the care of patients with advanced melanoma in the era before ipilimumab and may serve as a benchmark as new agents enter the melanoma treatment paradigm. These real‐world results are consistent with data from pivotal clinical trials conducted in an era when therapeutic options mirrored those available to physicians during our study. The majority of patients in this study had received prior systemic therapy, most commonly chemotherapy. We expect that the impact of these older treatments will not be tested after use of immune checkpoint inhibitors and/or molecular‐targeted agents in clinical trials, but evaluated instead in real‐world case series. Therefore, our data may be useful as a benchmark against which future clinical practice can be assessed. The conclusions that can be drawn from this analysis, however, are limited by the use of a pooled analysis from several countries , by the short follow‐up period (which may not completely reflect long‐term patient outcomes), and by prior therapy exposure . Despite these limitations, these results confirm the previous unmet need in advanced melanoma and provide historical information to facilitate the assessment of recent real‐world treatment patterns and trends in advanced melanoma.Mark R. Middleton, M.D., Ph.D.: Consulting/advisory role: GSK, BMS, Amgen, Merck, Roche , Clovis, Immunocore (uncompensated); research funding : GSK, AZ, Eisai, Clovis, BMS, Amgen, Roche, Merck, Vertex, Immunocore, Pfizer, Medimmune. Stéphane Dalle, M.D., Ph.D.: Research funding: Roche. Joel Claveau, M.D.: Amgen (consultant), BMS, GSK, Roche, Merck, Novartis (consultant and investigator for those five). Pilar Mut, M.D.: None. Sigrun Hallmeyer, M.D.: Speaker and consultant for BMS, consultant for TPI, and consultant for Cardinal Health. Patrice Plantin, M.D.: None. Martin Highley, M.D.: Attendance at advisory board for Merck. Srividya Kotapati, Pharm.D.: Employment: BMS; research funding: BMS. T. Kim Le, M.P.H., M.S.: Employment: BMS. Jane Brokaw, Ph.D.: Employment: BMS. Amy P. Abernethy, M.D., Ph.D.: Employment: Flatiron Health, Inc; research funding: BMS, DARA Biosciences, GSK, Celgene, Helsinn, Dendreon, Kanglaite, BMS, and Pfizer.


Over the past several thousand years, arid and semiarid China has experienced a series of asynchronous desertification events in its semiarid sandy and desert regions, but the precise identification of the driving forces of such events has remained elusive. In this paper we identify two rapid desertification events (RDEs) at ~4.6 ± 0.2 ka BP and ~3.3 ± 0.2 ka BP from the JJ Profile, located in the eastern Mu Us Sandy Lands. These RDEs appear to have occurred immediately following periods marked by persistently frequent and intense fires. We argue that such fire patterns, directly linked to an uncontrolled human use of vegetation as fuel, played a key role in accelerating RDEs by ensuring that the land surface was degraded beyond the threshold required for rapid desertification. This would suggest that the future use of a massive and sustained ecological program of vegetation rehabilitation should reduce the risk of destructive fire. More reliable modern research shows that RDEs during the past 10 ka occurred asynchronously in different sandy lands, or even different parts of the same sandy lands101012111678The arid and semiarid region of North China consists principally of sandy lands, deserts and loess deposits, and lies at the junction of four principal atmospheric circulatory systems . Under g1Here we report on a Mid to Late Holocene microcharcoal record used as a proxy for fire characteristics in the Mu Us Sandy Lands  in an atArtemisia, Caragana, Salixand Hippophae.The Mu Us Sandy Lands are surrounded by the Yellow River to the west, north and east, and are adjacent to the neighboring Chinese Loess Plateau (CLP) to the southeast . The rego44.594′N, 110o10.044′E; 1159 m above sea level) is located in the second terrace of the Tuwei River, a branch of the Yellow River, on the eastern edge of the Mu Us Sandy Lands. The profile is 740 cm deep; from the base upwards it is stratigraphically divided into paleosol, eolian sand, weakly-developed paleosol and gramineous layers . Additionally, four grain-size groups , and two shape types (sub-round (R) and sub-long (L)), were identified. Here, total MC   . The MCLn of 4.8 . The PC total MC .According to the assemblages, three distinct periods of fire strength versus vegetation cover are identified. Before 4.6 ± 0.2 ka BP (Stage I), the relation between fire and vegetation evinces a ‘bow’ shape pattern. Between 4.6 ± 0.2 ka BP and 3.3 ± 0.2 ka BP (Stage II), this relation exhibits a gentle, similarly increasing trend. After 3.3 ± 0.2 ka BP (Stage III), the relation between fire and vegetation follows a similar pattern to Stage II, though with a sharp increase in distances traveled by fires toward the end of the period .>50 μm/MC<50 μm ratio; the sub-long and sub-round shapes of grains are linked to vegetation types, and in particular grasses and woody plantsL>50 μm/MCR>50 μm ratio indicating a relatively greater proportion of grasses. At the same time, a higher PC can indicate dense vegetation cover (or vegetation mass) under a moister climate, especially when located under a single wind-blown sedimentary layer23As the residual matter from the incomplete burning of vegetation, microcharcoal is usually considered as the most reliable natural proxy for assessing the intensity and strength of fires161716171920etc.) in the JJ Profile further supported such a climatic pattern9During Stage I, the ‘bow’ shape of the vegetation cover versus fire strength time series correlated well with the reconstructed precipitation record based on pollen data from lakes Tianchi and Gonghai>50 μm/MC<50 μm, MCL>50 μm/MCR>50 μm and PC before the two RDEs exhibit particular characteristics ; and total PC decreases gradually from ~200 to ~20 grains·g−1.When looking at the records in more detail, total MC, MCeristics : both toA dry climate and frequent strong winds promote desertification. One classic explanation of the above results is the influence of nonlinear relations combined with feedback between desertification and the atmosphere122126122661410vis-à-vis millennial-scale oscillations282930However, through careful comparison, these two RDEs at 4.6 ± 0.2 ka BP and 3.3 ± 0.2 ka BP cannot be directly attributed to climate change, owing to the following four key contradictions. (i) These RDEs occur synchronously in the JJ Profile and at Daihai Lakei.e. the excessive burning of vegetation for fuel, and an accelerating land surface degradation beyond the RDE threshold. Humans exert a definitive impact upon the environment, as when they alter terrestrial vegetation patterns during the rapid development of agriculture, and when a sharp increase in the population necessitates significant changes in land use33We therefore propose that the aberrant patterns of fire intensity/strength exhibited prior to both RDEs are the direct result of human activity, http://www.ancient-origins.net). Covering over four square kilometers, this stone city, with numerous carved jade and mural fragments, indicates the existence of a very prosperous urbanized society at ~4 ka BP.The Mu Us Sandy Lands and their neighboring regions have been intensively inhabited since at least the Last Glacial Maximum3637388After considering the relations between RDEs, climate change and human activity in the Mu Us Sandy Lands, we outlined our RDE trigger models. Based simply upon the deterioration in the climate, the lands system moves towards a drier state until it reaches the RDE threshold ; this ca3638Today, the Mu Us Sandy Lands are an important agro-pastoral transition zone and a powerful ecologically-protective screen preventing the incursion of deserts from the south and east. According to this analysis, future major ecological rehabilitation should focus on improving vegetation coverage and avoiding fire hazards to reduce the risk of desertification. Once the vegetation has been seriously damaged, land degeneration will greatly increase the risk that the RDE threshold is crossed.How to cite this article: Miao, Y. et al. Human activity accelerating the rapid desertification of the Mu Us Sandy Lands, North China. Sci. Rep.6, 23003; doi: 10.1038/srep23003 (2016).


Fresh fruits of “Regina” cv. were characterized by the highest average content of phenolic acids and anthocyanins: 4.987 mg·g−1 and 0.636 mg·100 g−1. The pattern of mean contents of biologically active substances analyzed in air-dried fruits was similar. Significant differences in abilities to neutralize the DPPH radical to diphenylpicrylhydrazine by extracts made of examined wild strawberry fruits were also indicated.Chemical analyses carried out in 2011–2013 aimed at evaluating the contents of flavonoids, free phenolic acids, tannins, anthocyanins, and antioxidant activity (%) by means of DPPH radical neutralization ability in fresh and air-dried fruits of three wild strawberry cultivars. Examinations revealed differences in contents of biologically active substances determined in raw versus dried material depending on the cultivar. Mean concentrations of flavonoids and tannins were highest in raw fruits of “Baron von Solemacher” cv., which amounted to 1.244 mg·g Fragaria vesca L. is a well-known and valuable plant species; however, its cultivation is still much poorly spread [ Fragaria vesca are derived from a wild species, which can be found in woods and grasslands in Europe, Western Asia, North America, and temperate areas in Chile [ Fragaria vesca ssp. vesca have been cultivated for centuries in European gardens. Their widespread temperate growing range, self-compatibility, and long history of cultivation, coupled with selection for favorable recessive traits such as day neutrality, nonrunnering, and yellow-fruited forms, offer extensive genotypic diversity [Fragaria spp. is unusual in what is called the fruit actually originates from the expansion of the flower base (the receptacle) as a pseudocarp, with the real one seeded fruits (achenes) on the epidermal layer [Fragariae fructus) with unforgettable flavor are one of the raw materials achieved from wild strawberry [y spread . Currentin Chile . Fragariiversity . Despiteiversity . Their niversity , 4. Theriversity . Fragarial layer –7. Wild al layer –13. Red,rawberry .Small fruit breeding programs are currently used to acquire new cultivars improved for specific agronomic (yield and size), qualitative , and sensorial (colour and aroma) characteristics, all combined to increased disease resistance and plant adaptability. Nowadays, besides all these parameters, it is necessary to look for the specific bioactive components well known for their effect on human health. This aspect is now highly requested by the consumer . Flavono2 area (2.0 × 3.75 m) at 40 × 25 cm spacing  in three replicates. The agrotechnical experiment was carried out on dusty soil characterized by good abundance in nutrients and neutral reaction. Soil under wild strawberry cultivation was prepared according to commonly accepted recommendations applying manure (40 kg·ha) for the forecrop (onion). Phosphorus, potassium, and magnesium fertilizers were used before seedling planting at the following amounts: 80 kg·ha—P2O5; 100 kg·ha—K2O. Starter rate (N—30 kg·ha) was applied, when the seedling was taken the roots. The wild strawberry plantation was regularly manually deweeded. First harvest was in second growing years. The fruit harvest was carried out at the full of fruiting stage in years 2011–2013. Fruits were harvested once in the early morning, in June every year.The study material consisted of fruits collected from three cultivated wild strawberry cultivars: “Baron von Solemacher,” “Yellow Wonder,” and “Regina” originating from agrotechnical experiments carried out at Department of Vegetable and Medicinal Plants, University of Life Sciences, Lublin . Seeds of tested cultivar were sown manually on March 5th, 2010, into boxes filled with a substrate (peat substrate) and covered with thin sand layer. After emergence and forming 2-3 true leaves, the seedlings were transferred into boxes at 5 × 3.5 cm spacing. Plants were set into their permanent place on June 20th, 2010, in plots of 7.5 m Fragaria vesca are presented in Weather conditions during growth and studies upon−1) [−1) [−1) [Directly after the harvest, part of material was subject to laboratory analyses as raw, while another part was dried out. The drying process was performed in a drying facility at 40°C till the moment of a constant air-dried fruit weight achievement. Raw material was subject to determinations of dry matter (%) by means of drier method , weight −1) , sum of −1) [−1) , tannins−1) [−1) , anthocy−1) [−1) , and ant−1) [−1) . BiochemDry matter determination was carried out by means of drier method according to Charłampowicz . AliquotThe loss after drying was determined by means of gravimetric method according to Polish Pharmacopoeia VII . Samples−1), and 1 mL of methenamine (5 g·L−1) were then added and the mixture was maintained for 30 min under reflux on a water bath. The hydrolysate was filtered through cotton wool into a volumetric flask of 100 mL, then placed in a flask together with the cotton pellet and 20 mL of acetone, and refluxed for 10 min. Next, 20 mL of solution was dispensed into a separatory funnel with 20 mL of water and extracted with ethyl acetate in 15 mL portions 3 times with 10 mL. The combined organic layers were washed twice with 40 mL of water, filtered into a volumetric flask of 50 mL, and supplemented with ethyl acetate. To determine flavonoid content, 2 samples were prepared: to 10 mL of the stock solution 2 mL of a solution of aluminum chloride (20 g·L−1) was added, supplemented with a mixture (1 : 19) of acetic acid (1.02 kg·L−1) and methanol (25 mL). To prepare the comparative solution, stock was supplemented with 10 mL of a mixture (1 : 19) of acetic acid (1.02 kg·L−1) and methanol (25 mL). After 45 min, the absorbance of the solutions was read at λ = 425 nm on HITACHI U-2900 spectrophotometer using the reference solution for comparison. Samples were analyzed in 3 replicates. The total content of flavonoids (mg·g−1) was expressed as quercetin QE equivalent according to the following formula:X are total flavonoids (mg·g−1); A is the absorbance of the solution being studied; k is the convection factor for quercetin and equal to 8.750; m is the sample with the raw material (g) which was the amount of fresh and dry material.Total flavonoids were estimated according to the spectrophotometric method of Christ and Müller  after thλ = 490 nm. The total phenolic acid content, expressed as acid equivalent weight of caffeic acid (CAE) in the fruit, was calculated from the equation obtained from the calibration curve of caffeic acid . Samples were analyzed in triplicate.Total phenolic acids estimation was carried out according to Arnov method , which cThe amount of tannin estimation was determined using Pharmacopoeia procedure . The con3) and heated in water bath for 1 hour. The obtained extract was hydrolyzed with 20 mL n-butanol, and then two portions of 10 mL n-butanol were added as a solution. Anthocyanin extract was rinsed in 50 mL flask with n-butanol. The absorbance was measured immediately at 533 nm [P are total anthocyanins (mg·100 g−1); A is absorbance at 533 nm; V is value of butanol phase (50 mL); F is coefficient for delphinidin chloride ; m is mass of sample to be examined (mg).Samples of raw material (1.0 g) were extracted with 50 mL HCl (1 mol·dmt 533 nm . The perμg·mL−1 concentration. The absorbance measurements were made at λ = 517 nm wavelength using spectrophotometer HITACHI U-2900.Antioxidant activity (%) was evaluated on a base of the ability to neutralize the DPPH radicals by means of spectrophotometry according to Chen and Ho : to do tAll reagents and solvents were of analytical grade chemicals from Merck  or Sigma Chemical Co.  and POCH .Achieved results from laboratory experiments were statistically processed by means of variance analysis method and Tukey's confidence intervals at 5% confidence level.Determinations related to the chemical composition of three wild strawberry cultivars fruits here presented were preceded with the evaluation of general physicochemical parameters, that is, dry matter content in raw fruits and weight loss after drying-moisture content in dried fruits. Data presented in Among compared cultivars, “Baron von Solemacher” fruits were characterized by the highest concentration of the component , while “Yellow Wonder” contained the lowest level of the parameter (28.22%). Significant differences between cultivars in subsequent years of study were observed. Considering the water content in dried material, fruits of studied wild strawberry cultivars slightly differed from each other and statistical analysis did not reveal any significant differences. Regardless of the cultivar, moisture content of air-dried fruits oscillated around 11.10%, on average .−1  and 1.245 mg·g−1 . On the other hand, the lowest quantities of analyzed compounds were recorded in fruits of Yellow Wonder cv. . Significant differences in flavonoids contents over the years of study for both analyzed types of material were observed. The dried fruits of all studied genotypes were characterized by over twice as high flavonoids amounts as compared to raw fruits.Different levels of flavonoids in raw and dried fruits of three wild strawberry cultivars are presented in −1 in raw and 4.987 mg·g−1 in dried material  and from 192.52 mg·100 g−1 to 236.90 mg·100 g−1  of anthocyanins. The lowest levels of analyzed substances for all studied genotypes were recorded in 2012.Anthocyanins were another group of substances analyzed in wild strawberry fruits . Fruits Significant differences in the ability to neutralize the free DPPH (diphenylpicrylhydrazyl) radical by extracts made of examined wild strawberry fruits were recorded .Extracts prepared from dried fruits revealed definitely highest ability (23.93%) as compared to raw material (13.11%). When comparing studied cultivars, the highest neutralizing capacity was shown by extracts made of Regina cv. fruits 14.27%  and 24.60% (dried fruits). The free radicals were worse reduced by extracts prepared from fruits of Yellow Wonder cv. and Baron von Solemacher cv.Fragaria vesca has traditionally been a popular delicious fruit for its flavor,taste, fresh use, freezing, and processing. Morphological, biometric, and agronomical characteristics have been widely used to describe wild strawberry cultivars. In recent years, cultivated berries have become very attractive for consumers because of potentially beneficial phytochemicals contained in these fruits. The importance of flavonoids and other phenolics has been suggested to play a preventive role in the development of cancer and heart disease [ p-coumaric, gallic acids), flavonoids , proanthocyanidins, and anthocyanins  [ disease . Conside disease , 25. Folyanidin) –28. Due yanidin) , 30. Meayanidin) . However−1 and 6.09%, respectively. Raw fruits of “Regina” cv. were characterized by the highest average concentrations of phenolic acids and anthocyanins: 4.987 mg·g−1 and 444.25 mg·100 g−1, respectively. According to Antal et al. [−1 (Fragaria moschata) to 165 mg·100 g−1 (Vaccinium myrtillus). Huang et al. [ Fragaria x ananassa Duch. genus. Antal et al. [−1. The differences in polyphenol content of strawberry fruit from the literature may be due to the different conditions during the growth of plants , the length of the growing season, and harvest date. They may also be caused by using various analytical procedures or methods identifying the active ingredients by scientists in various centers. Identification of chromatographic methods usually has a lower content of active substances, because its task is to determine the minimum content and separation and identification of individual compounds in the raw material. Commonly used spectrophotometric procedures are based on similar assumptions, so after taking into account differences methods are comparable to those applied in this paper. During the three years of the research, the highest contents of analyzed biologically active substances were recorded in 2013, while the lowest were recorded in 2012.Secondary metabolites content in berry fruits varies among species and cultivars, but it can also be affected by growth conditions including environmental factors and cultivation techniques , 31–34. l et al. , the quag et al.  found hig et al. , the amol et al.  evaluateIn a previous investigation, we found that fresh wild strawberry fruits possess high amounts of bioactive compounds. However, fresh fruits are not available all year round, being harvested in Poland only in June–September. Therefore, it is important to find a proper substitute that could be used when fresh berries are not available. It was decided to prepare dried fruits, to determine the contents of some important bioactive compounds and their antioxidant potential therein, and to compare them with the same parameters in fresh fruits. Dried fruits have a greater nutrient density, greater fiber content, increased shelf life, and significantly greater phenol antioxidant content compared to fresh fruits. The quality of the antioxidants in the processed dried fruit is the same as in the corresponding fresh fruit. Phenols in dried fruit may be important antioxidants . Data prAs a conclusion, our results clearly demonstrate that considerable variation exists in the phenolic compounds among wild strawberry genotypes. The obtained results allowed the identification of cultivars which were characterized by the highest content of investigated biologically active substances and the highest ability to free radicals elimination. Fruits of these cultivars were characterized by the highest prohealth properties. The results from the performed experiments also have practical application during the breeding program preparation in order to obtain new cultivars of this species. Therefore, dried fruits are good source of important bioactive compounds and more dried fruits should be recommended to be added to the diet by dieticians and nutritionists.


Background: Serum lutein (L) and zeaxanthin (Z) positively correlate with macular pigment optical density (MPOD); hence, the latter is a valuable indirect tool for measuring L and Z content in the macula. L and Z have been attributed antioxidant capacity and protection from certain retinal diseases but their uptake within the eye is thought to depend on genetic, age, and environmental factors. In particular, gene variants within beta-carotene monooxygenase (BCMO1) are thought to modulate MPOD in the macula.Objectives: To determine the effect of BCMO1 single nucleotide polymorphisms (SNPs) rs11645428, rs6420424, and rs6564851 on MPOD in a cohort of young healthy participants of Caucasian origin with normal ocular health.Design: In this cohort study, MPOD was assessed in 46 healthy participants  with a mean age of 23.8 ± 4.0 years (range 19–33). The three SNPs, rs11645428, rs6420424, rs6564851 that have established associations with MPOD were determined using MassEXTEND (hME) Sequenom assay. One-way analysis of variance was performed on groups segregated into homozygous and heterozygous BCMO1 genotypes. Correlations between body mass index (BMI), iris color, gender, central retinal thickness (CRT), diet, and MPOD were investigated.Results: Macular pigment optical density neither significantly varied with BCMO1 rs11645428 , rs6420424  nor rs6464851 homozygous or heterozygous genotypes , in this young healthy cohort. The combination of these three SNPs into triple genotypes based on plasma conversion efficiency did not affect MPOD . There was a significant negative correlation with MPOD and CRT  but no significant correlation between BMI, iris color, gender, and MPOD.Conclusion: Our results indicate that macular pigment deposition within the central retina is not dependent on BCMO1 gene variants in young healthy people. We propose that MPOD is saturated in younger persons and/or other gene variant combinations determine its deposition. L), zeaxanthin (Z), and meso-zeaxanthin (MZ) and has protective functions including those as an antioxidant and short-wavelength (blue) light filter within the central retina  are all positively correlated in healthy samples  to determine whether the single SNP genotypes or their combinations can explain the variation in MPOD in a younger cohort.The critical role of gene variants in the determination of MPOD has only recently received attention among the scientific community –18, with1 action , 22 withntration . There iand MPOD , 17 but enotypes . The objParticipants were recruited in the study according QUT’s human ethics approval. Participants were included if they were aged between 19 and 33 years, female or male, of normal general and ocular health and excluded if they had a color vision deficiency or were taking supplements containing L or Z. Of the 48 participants who volunteered, one was excluded due to protanopic congenital color vision deficiency and one due to zeaxanthin supplementation. This resulted in a sample of 46 participants eligible for further testing. All recruitment and experiments were conducted in accordance with the QUT Human Research Ethics Committee  and the tenets of the declaration of Helsinki.To determine the general health and well-being of the study cohort the familial and personal medical history was taken and the BMI was calculated . BecauseA macular pigment densitometer  was used to perform heterochromatic flicker photometry (HFP). The protocol followed the standardized protocol for measuring MPOD by HFP  and as pBCMO1 gene of each participant, a 2 mL saliva sample was collected with Oragene™ self-collection kits . Samples were stored at −80°C before manual DNA purification, which was performed according to the manufacturer’s protocol and Australian Genome Research Facility (AGRF)  guidelines. BCMO1 SNPs rs11645428 (SNP1), rs6420424 (SNP2), and rs6564851 (SNP3) were genotyped using a commercial genotyping service provided by the AGRF. The method used employed matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to genotype using a homogenous MassEXTEND™ assay .To determine the SNPs within the post hoc analysis where necessary, were performed. In addition, a Pearson correlation was performed to determine the relationship between MPOD, gender, ethnicity, central retinal thickness (CRT), BMI, and iris color. A p-value of 0.05 was considered as statistically significant.All statistical analysis was performed using the software SPSS version 20 . Initial screening indicated all data met the assumptions of the statistical tests. To determine whether there was a significant difference between the three SNP genotypes, either singly or in combination, and MPOD, a one-way analysis of variance (ANOVA) and Genotype frequencies were determined for rs11645428, rs6420424, and rs6564851 . For the BCMO1 SNP rs6420424, the mean MPOD for the homozygous A allele was 0.50 ± 0.2 D.U., the homozygous G allele was 0.51 ± 0.2 D.U., and the heterozygous GA allele, 0.47 ± 0.2 D.U. . The mean MPOD for the homozygous T, G, and heterozygous GT allele for the BCMO1 SNP rs6564851 were 0.50 ± 0.2 D.U., 0.49 ± 0.2 D.U., and 0.47 ± 0.2 D.U., respectively .For BCMO1 SNPs on MPOD, we investigated triple genotype combinations based on BCMO1 plasma conversion efficiency as it has been investigated in a mixed cohort of participants over the age of 50 years .While there was no significant effect of single 50 years . The tri50 years , which wp = 0.3), BMI (p = 0.7), iris color (p = 0.8), and smoking status (p = 0.5) whereas CRT was negatively correlated . The results of 36 participants who completed the dietary questionnaire demonstrated that participants with the lowest dietary score  had on average a lower MPOD (0.30 D.U. ± 0.21) compared to those participants with the highest score  who had an MPOD of 0.55 D.U. ± 0.13. The remaining participants with scores 2 (n = 8), 3 (n = 15), and 4 (n = 5) had an average MPOD of 0.5 D.U. ± 0.15, 0.46 D.U. ± 0.15, and 0.48 D.U. ± 0.1, respectively.A Pearson correlation showed that MPOD was not significant correlated with gender , there was no significant difference in MPOD between the three BCMO1 mediates macular pigment uptake. The observation that BCMO1 SNP genotypes correlated with MPOD variation in older healthy participants but not in the younger cohort in the current study, indicates that age may have an effect on macular pigment transport and/or deposition mechanisms.Feigl et al.  demonstrBCMO1 SNP rs11645428 was correlated with baseline MPOD prior to supplementation. However, after supplementation of L and Z, this polymorphism was no longer significantly affecting MPOD. This may indicate that the threshold for a BCMO1 effect on MPOD had been surpassed with the increased levels of L and Z, similar to a young cohort with a healthy diet as apparent in our current study.The point of MPOD saturation or maximal MPOD in a given cohort has been observed in numerous supplementation studies , 27 but L and Z accumulate in the macula, serving as a “sink”  that were previously associated with carotenoid metabolism and MPOD  the SNP rs838879 affected MPOD in such way that participants with the homozygous G allele had on average higher MPOD compared to the homozygous A and heterozygous GA allele. The number of participants with the GG genotype, however, was low and further studies are needed to confirm whether SCARB1 rs838879 GG is associated with higher MPOD.The carotenoids a “sink”  to the ea “sink” –38 have a “sink” , 18, 39.a “sink” . In partn uptake . We alsoand MPOD , 17, 41.We found a significant negative correlation between CRT and MPOD as previously reported  with higBCMO1 SNP genotypes do not explain variations in MPOD, whereas they have previously been shown to play a role in MPOD in older participants (In summary, this is the initial demonstration that in a mixed gender cohort of young participants aged ≤33 years, icipants . We propThe 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.


Elsholtzia rugulosa, a shrub species in southwestern China. We monitored the pollinating honey bee Apis cerana, as well as two occasional visitors – Vespa velutina and a Bombus sp. – and an herbivorous beetle Oxycetonia jocunda. Behavior experiments using Y-tubes showed that honey bees were attracted primarily by floral scent while hornets and bumble bees were repelled by both the flowers and leaves. Analysis via gas chromatography-mass spectrometry revealed that these tissue types differed in their compositions and relative amounts of volatile compounds. When the plants were damaged, the average relative amounts of Elsholtzia ketone rapidly increased in the flowers and leaves. Furthermore, herbivorous beetles were strongly repelled by damaged tissues, suggesting a potential defense signaling function by these compounds. Our findings again demonstrate that scents have multiple functions in the interactions among plants and insects.Although the roles of volatile compounds have been examined separately in plant–herbivore or plant–pollinator interactions, few studies have focused on how plant scents can attract effective pollinators, repel ineffective pollinators, and defend against attacks by insect herbivores. We explored the functional significance of volatile compounds that impart a strong odor to  Some flowering plants bring pollinators into contact with reproductive structures by providing olfactory or visual signals, and by rewarding visitors with nectar, pollen, and oils23567891112Hemipepesis (Hymenoptera)12171819Volatile compounds help plants communicate and interact with their surrounding environment. These volatiles, released from leaves, flowers, and fruits into the atmosphere and from roots into the soil, provide a reproductive advantage by attracting pollinators and seed dispersers121314Elsholtzia rugulosa have a strong odor and are medicinally important in ChinaApis cerana) and seed production increases in parallel with the number of pollinator visits to attain higher yields. Field observations have shown that hornets (Vespa velutina) and bumble bees (Bombus sp.) are also occasional floral visitors to these flowers (unpublished data). We visually confirmed that their heads and bodies did not make effective contact with the anthers and stigmas during visiting flowers occasionally, the longer tongues of hornets and bumble bees were unsuitable for pollination of E. rugulosa. At the study site, the few hornets and bumblebees caught in the mist net were lacking E. rugulosa pollen. No reports have been made on either the constitution of the unusual sharp odor in leaves and flowers of E. rugulosa or the significance of its function. However, whole-plant studies have examined the chemical components of its essential oilsPlants of Apis cerana bees significantly preferred the branch containing Elsholtzia rugulosa flowers over the empty control branch     . They al n = 42) . However n = 35) .By contrast, the hornets and bumblebees were strongly repelled by the branches containing flowers or leaves over the empty control branch , and theP = 0.05; leaves, 24.76 ± 4.96, P = 0.02). Principal component analysis demonstrated that the first two components explained 65.64% and 19.39% of the total variation .Apis cerana (n = 37), occasional visitors Vespa velutina (n = 18) and Bombus sp. (n = 15), and beetles (n = 20) were captured and kept in individual containers (20 cm2) for up to 20 min prior to the start of the experiments. The insects were placed at the entrance and allowed to walk down the Y-tube to select one of the branches.To test the importance of floral scent as an attractant, we performed insect behavior experiments in the laboratory with a glass Y-tube 12−1 with a flow meter. Sampling periods were 3 to 4 h long. Empty cooking bags placed in close proximity to the inflorescences were used as controls. After the fragrance sampling was completed, adsorbed volatiles were eluted from the Super-Q with 1.5 mL of dichloromethane , then sealed in glass vials and stored at –20 °C. Scent samples were collected during the sunniest time of the day, between 12:00 and 16:00, which coincided with when the floral visitors normally are present. Leaves were damaged in a manner that mimicked beetle attack in the fieldScents from intact and damaged inflorescences, and from normal and damaged leaves, were collected outdoors by methods utilizing dynamic headspace adsorption−1. The GC oven temperature began at 50 °C and was increased by 5 °C min−1 to 100 °C, held for 10 min, then increased by 5 °C min−1 to 280 °C and held for 5 min. The MS interface was 280 °C and the ion trap was activated at 150 °C. Mass spectra were taken at 70 eV (in EI mode) with a scanning speed of 1 per scan from m/z 35 to 550. All compounds were compared against the Wiley NIST 05 mass spectral database and Wiley 7.The volatiles were analyzed on a Hewlett-Packard 6890 Series GC System coupled to a Hewlett-Packard 5973 Mass Selective Detector and an Agilent 7683 Series Automatic Liquid Sampler. An HP-5MS column  was used. Constant helium gas flow was controlled electronically at 1.4 mL minThe results from each behavioural experiment were compared using a binomial test to establish if wasps showed overall preference. All analyses were performed using SPSS 16 , with measured variables presented as mean ± SE.How to cite this article: Zhang, F.-P. et al. Multiple functions of volatiles in flowers and leaves of Elsholtzia rugulosa (Lamiaceae) from southwestern China. Sci. Rep. 6, 27616; doi: 10.1038/srep27616 (2016).


Bistorta vivipara bulbil ecotypes from a subarctic elevational gradient (temperature range: ±3°C) in northern Sweden to disentangle effects of local ecotype, temperature, and biotic and abiotic properties of soil origin on plant growth. We found partial evidence for local adaption to temperature. Although soil origin affected plant growth, we did not find support for local adaptation to either abiotic or biotic soil properties, and there were no interactive effects of soil origin with ecotype or temperature. Our results indicate that ecotypic variation can be an important driver of plant responses to the direct effects of increasing temperature, while responses to covariation in soil properties are of a phenotypic, rather than adaptive, nature.Predicting how plants will respond to global warming necessitates understanding of local plant adaptation to temperature. Temperature may exert selective effects on plants directly, and also indirectly through environmental factors that covary with temperature, notably soil properties. However, studies on the interactive effects of temperature and soil properties on plant adaptation are rare, and the role of abiotic versus biotic soil properties in plant adaptation to temperature remains untested. We performed two growth chamber experiments using soils and  Other studies have suggested that plant adaptation to temperature may trade-off against adaptation to insect herbivory. Notably, plants growing at higher elevation would need to allocate fewer resources to herbivore defence because the lower temperatures would moderate herbivore pressure [Local adaptation of plants to environmental conditions is receiving increasing interest, particularly in the context of climatic change ,2. Pattepressure .Few studies, however, have considered interactions with belowground factors ,7 and we°C, which makes this system powerful for comparisons with predicted future climate change scenarios [We used an elevational gradient in subarctic Sweden to test local plant adaptation to temperature and soil properties. Previous studies in this system have shown that an elevational increase of approximately 500 m corresponds with a decrease in growing season temperature of about 3cenarios . Along tcenarios , which ccenarios , with pocenarios ,15. To fBistorta vivipara, a viviparous, perennial, bulbil-producing plant species commonly found in (sub)arctic and alpine habitats [B. vivipara bulbils were collected from three elevations (i.e. three temperature habitats) and seedling responses were tested in two fully reciprocal experiments , using climate-controlled growth chambers to mimic conditions of each temperature habitat. We tested the following hypothesis: local adaptation of B. vivipara to temperature is coupled with adaptation to associated soil properties, due in part to co-adaptation of plants and soil organisms. To diagnose local adaptation, we used two non-independent criteria [We tested patterns of local adaption for habitats . Soils acriteria ,2: (i) t3.3.1B. vivipara (syn: Polygonum viviparum) bulbil ecotypes used in this study were collected from an elevational gradient on the northeast facing slope of Mount Suorooaivi (1193 m), approximately 20 km southeast of Abisko, Sweden  [°C, 12.3°C and 9.7°C, respectively; the average minimum temperature corresponded to 5.0°C, 4.1°C and 3.8°C (a). Mean annual precipitation is approximately 310 mm . Parent soil material is composed of salic igneous rocks and quartic and phyllitic hard schists. The vegetation typically consists of mosaics of two co-dominant types: meadow and heath, which occur across the gradient. Meadow vegetation is diverse and dominated by forbs, graminoids and sedges, while heath vegetation is less diverse and dominated by deciduous and evergreen dwarf-shrubs and Betula nana. Bistorta vivipara typically occurs in meadow vegetation [Soils and 8°49′ E) ,15. The nd 3.8°C a. Mean a3.2ca 48 m with the maximum distance between the two plots furthest apart being ca 140 m. Because of high spatial heterogeneity over short distances in micro-topography, hydrology and soil fertility in these plant communities [°C) for six weeks until they were further processed for our experiments; these freezing conditions are well within the range of what these soils experience during the winter.Soils for our experiments were collected in August 2012 at 450, 700 and 900 m elevations. Soils were collected to 10 cm depth adjacent to five existing 1×1 m meadow plots at each elevation established earlier in 2012. The mean distance between plots within elevations was munities , it is emunities . After c3.3°C overnight before being passed through a 4 mm mesh sieve to remove plant matter and stones. Soil pH was determined on a subsample of fresh soil (2.5 g dry weight) after shaking for 12 h in 40 ml deionized water. Gravimetric moisture content was determined after drying  and soil organic matter content was determined after combustion in a muffle furnace . A subsample of soil was dried , ground and analysed for total carbon (C) and nitrogen (N) by dry combustion using a FLASH 2000 Organic Elemental Analyzer  and phosphorus (P) with nitric–perchloric acid digestion analysed by inductively coupled plasma [et al. [ω9, 16 : 1ω7c, 16 : 1ω7t, i17 : 0, a17 : 0, 17 : 1ω8, cy17 : 0, 17 : 0, 18 : 1ω7, cy19 : 0 as indicators for bacteria and 18 : 2ω6 as an indicator for fungi.In July 2012, we collected three to six soil cores from each of the five 1×1 m meadow plots at each elevation  for analysis of abiotic and biotic soil properties. Cores were bulked within plots and kept at 4d plasma . For eacd plasma  with dif [et al. . We used3.4Bistorta vivipara is a viviparous perennial knotweed commonly found in (sub)arctic and alpine habitats [B. vivipara normally reproduces asexually by bulbils (vegetative reproduction organs), and this should serve to limit gene flow. Notably, the potential for local adaptation is highest for ‘intermediate’ levels of gene flow [Bistorta vivipara clones can be widespread [B. viviparapopulations in arctic and alpine zones is rather high, with many clones being unique to single populations [B. vivipara a good candidate species to address our hypothesis. At each elevation, B. vivipara bulbils were collected within 25 m of the plots. Bulbils were collected between mid-August and early September 2012 from approximately 200 individuals per elevation. These bulbils were stored at 4°C for a maximum of six weeks, after which they were surface-sterilized (1% sodium hypo-chloride solution for 1 min) [habitats ,23–25. Vhabitats ,27, and habitats ,28, but ene flow ,30. Bistulations ,31,32. Ar 1 min) , and ger3.5B. vivipara to temperature is coupled with adaptation to associated soil properties. This experiment included three fully crossed factors: soil origin , bulbil ecotype  and temperature , with five replicates of each treatment combination . Soils were processed by removing large stones and roots and then homogenized per elevation, because we were interested in local adaptation to soil properties related to elevation, rather than small-scale within-site heterogeneity [B. vivipara seedlings.The soils and bulbil ecotypes were then used in two growth chamber experiments. Experiment 1 addressed whether local adaptation of ogeneity . For eac°C for the 450 m elevation, 4.1 and 12.3°C for the 700 m elevation, and 3.8 and 9.7°C for the 900 m elevation. Weekly average maximum and minimum temperatures were derived from equations of the polynomial trend lines of regressions as shown in figure 1a. Diurnal temperature cycles in the growth chambers were set by hourly increments between minimum and maximum values. Minimum temperature values lower than 5°C were set to 5°C owing to climate chamber system constraints. Relative humidity was set to 90% in all chambers, and photosynthetically active radiation (PAR) and hours of daylight were set in each growth chamber to represent conditions measured from mid-June to mid-September 2012 in the vicinity of the elevational gradient used in our study . Each of the five replicates of each treatment combination was randomly assigned to a position in a block within each chamber. All pots were isolated in individual saucers to prevent cross-contamination through water movement. Pots were rotated systematically between chambers (i.e. entire temperature treatments moved between chambers) on a weekly basis to minimize effects of any uncontrolled sources of variation among chambers. Pots were watered as necessary throughout the experiment to eliminate moisture as a limiting factor.For each soil origin × ecotype combination, five replicate pots were placed in each of three growth chambers, whose conditions were set to mimic temperature conditions as observed at 450, 700 and 900 m along the elevational gradient during the 2012 growing season . Averageγ-irradiation (25 kGy), which is widely used for killing soil biota [B. vivipara seedlings as described for experiment 1. The experiment was then run following approaches and methodology described for experiment 1.Experiment 2 addressed whether local adaptation to temperature is due in part to co-adaptation of plants and soil organisms; it differentiated between the effects of abiotic and biotic soil properties using sterilized soils and living soil inocula. By comparing sterilized soils to reinoculated soils, the effects of abiotic versus biotic soil factors on plant growth can be effectively separated ,35. The il biota . We prep°C for at least 72 h.For both experiments, plants were grown for 12 weeks, following temperature and light conditions characteristic for the Abisko growing season (3.6n=5). To assess significant differences among means, Tukey's HSD post hoc tests with Bonferroni corrections (p=0.05) were run for (i) ‘local versus foreign’ comparisons, and (ii) ‘sympatric versus allopatric’ comparisons [post hoc tests. For all data analyses, data were transformed when necessary to meet the assumptions for parametric testing. All statistical analyses were performed in SPSS .The effects of ecotype, temperature and soil/inoculum origin, and their interactions, on total plant biomass were tested using three-way ANOVA. Biomass of the two seedlings per pot was averaged, and pots served as the unit of replication . Local versus foreign contrast (figure 2a): at the 450 m temperature, biomass of the 450 m ecotype was higher than for the 700 and 900 m ecotypes; at the 700 m temperature, biomass of the 450 and 700 m ecotypes was higher than the 900 m ecotype; and at the 900 m temperature, biomass of the 450 m ecotype was higher than the 900 m ecotype. Sympatric versus allopatric contrast : for the 450 and 700 m ecotypes, biomass was highest at temperature treatments corresponding to their elevation of origin; and there was no temperature effect on the ecotype from 900 m.There was a significant interactive effect between ecotype and temperature on plant biomass, but there were no interactions of soil origin with ecotype or temperature . Furtherfigure 24.2c): at the 450 m temperature, the 450 m ecotype had significantly higher biomass than did the 700 and 900 m ecotypes; similarly, at the 700 m temperature, the 700 m ecotype had a significantly higher biomass than did the two other ecotypes; and at the 900 m temperature there were no significant biomass differences between the three ecotypes (b). Plant biomass was not affected by inoculum origin, or by its interaction with ecotype and/or temperature (d). Sympatric versus allopatric contrast : for the 450 and 700 m ecotypes, biomass was highest at temperature treatments corresponding to their elevation of origin; and there was no temperature effect on the ecotype from 900 m. Biomass of the 450 and 700 m ecotypes differed significantly from that of the 900 m ecotype, but not from each other.Plant biomass patterns in experiment 2 were generally similar to those observed in experiment 1. Local versus foreign contrast c: at theecotypes b. Plant perature d. Sympat5.B. vivipara to shifting conditions with elevation, largely manifested by adaptation to temperature. Ecotypes from the lowest elevation performed best at local temperatures , and at temperatures representing low elevation the local ecotype performed best . Similar patterns were found for the mid-elevation ecotype and temperatures. These patterns could not solely be explained by the more favourable temperature conditions at lower elevations . Notabl limited ,24,39. MContrary to our hypothesis, we found that plant adaptation to temperature was independent of soil conditions. Overall, plants performed best in low elevation soils, probably because of increased soil fertility at lower elevations ,40. HoweB. vivipara to changing future temperatures would largely depend on the genetic variability in temperature optimum, and would not be impeded by soil legacies. As such, our study sheds new light on the relative importance of genetic adaptation and phenotypic plasticity for plant performance under shifting climatic conditions.Understanding local adaptation of plants to environmental conditions is important in predicting plant performance and population dynamics under future climate change scenarios . Our res