Body Mass Index (BMI), like most human phenotypes, is substantially heritable. However, BMI is not normally distributed; the skew appears to be structural, and increases as a function of age. Moreover, twin correlations for BMI commonly violate the assumptions of the most common variety of the classical twin model, with the MZ twin correlation greater than twice the DZ correlation. This study aimed to decompose twin correlations for BMI using more general skew-t distributions.N = 7,086) from the community-based Washington State Twin Registry were included. We used latent profile analysis (LPA) to decompose twin correlations for BMI into multiple mixture distributions. LPA was performed using the default normal mixture distribution and the skew-t mixture distribution. Similar analyses were performed for height as a comparison. Our analyses are then replicated in an independent dataset.Same sex MZ and DZ twin pairs (A two-class solution under the skew-t mixture distribution fits the BMI distribution for both genders. The first class consists of a relatively normally distributed, highly heritable BMI with a mean in the normal range. The second class is a positively skewed BMI in the overweight and obese range, with lower twin correlations. In contrast, height is normally distributed, highly heritable, and is well-fit by a single latent class. Results in the replication dataset were highly similar.Our findings suggest that two distinct processes underlie the skew of the BMI distribution. The contrast between height and weight is in accord with subjective psychological experience: both are under obvious genetic influence, but BMI is also subject to behavioral control, whereas height is not. Body Mass Index (BMI) is one of the more intensively studied phenotypes in the genetic epidemiology literature. A 2012 meta-analysis identified 88 individual effect sizes from twin studies . First, The primary goal of this study was to use latent profile analysis (LPA) to decompose the twin correlations and corresponding biometric components for BMI into multiple distributions using a large sample of twins. LPA estimates statistical models in multiple classes determined by the structure of the data. Our analysis differs from typical applications of LPA to twin studies in several ways. First, it has recently become possible to loosen the classical method of decomposing variables into normally distributed latent profiles (or classes); recent developments using the software Mplus allow dependent variables to be decomposed into more general skew-t distributions , making Second, the usual strategy in LPA-based models of twin studies has been to estimate the classes on the individual phenotypes, and subsequently estimate the twin correlations for the estimated classes , 11. HerThe study was approved by the IRB at the University of Virginia (SBS #2014036900).This study included a sample of 7,086  twin pairs from the community-based Washington State Twin Registry within a cross-sectional study design. Twins include same-sex male and female twin pairs aged 18–97 years, reared together. Participants were recruited from Washington State driver’s license and identification card applications . All twikg/m2. These measures were collected from responses to the survey questions “What is your current height?” in feet and inches and “What is your current weight?” in pounds. In a sample of twins (n = 144 pairs) participating in an ongoing funded study  of 0.70 and 0.58 for men and women; a small proportion of the variance attributed to the shared family environment  and non-shared environment and measurement error (1 - rMZ = 0.29 and 0.27 for men and women). These findings are consistent with previous reports of twin studies of BMI [We began by estimating twin correlations and a classical twin (ACE) model for BMI in men and women. In men, the MZ twin correlation , indicating that excessive skewness and kurtosis need to be taken into account when modeling the distribution of BMI. For both men and women, the two-class skew-t distribution mixture models had lower BICs than the one- and three-class skew-t mixture models, suggesting that the two-class solutions were better fit for both gender. The entropy value probabilities were 0.723 and 0.783 for men and women, respectively, suggesting adequate latent class separation (18). For men, the three-class skew-t distribution mixture model had zero individuals in one of the latent classes, suggesting that a third latent class was not needed. Similar results were found for women: one of the latent classes had less than 2% of the participants, suggesting that the three-class skew-t distribution mixture model was not a good fit.The fit statistics of the mixture models under normal and skew-t distributions for BMI are presented in M ~ 22kg/m2 for both men and women), and a skewed distribution, with mean BMI in the overweight range (M ~ 29kg/m2 for both men and women). We subsequently labeled the latent profile with mean BMI in the normal range as the "normal" class, and the other one the "overweight" class.The number of individual twins in each latent class stratified by their measured BMI is presented in rMZ = 0.79 and rDZ = 0.28 for men; rMZ = 0.71 and rDZ = 0.29 for women) are indicative of high heritability, no effect of family environment, a relatively small effect of non-shared environment, and a modest violation of the ACE model. In contrast, the smaller twin correlations in the overweight class  suggest a large non-shared environmental effect and a severe violation of the ACE model. These twin correlations are illustrated in The twin correlations in the normal class  than those in the normal class (mean age = 34 to 40 years old).df = 5.80, SE = 0.51, p < .0001 for men; df = 6.27, SE = 0.47, p < .0001 for women) deviated from that of the normal distribution (df ≥ 30), whereas the skew parameters were non-significant . These results suggested the need to account for excessive kurtosis but not excessive skewness for height. Subsequent multi-class mixture models were only estimated using normal distributions.The fit statistics of the mixture models under normal and skew-t distributions for height are presented in For both men and women, although the two-class mixture models showed improvements in BIC over the one-class mixture model, more than 90% of the participants were estimated to belong to one class (94% for men and 95% for women), suggesting that the one-class solution was a sufficient fit to the current data for both men and women. Descriptive statistics of height by sex and zygosity are presented in The twin correlations of height by sex and zygosity are also illustrated in SD = 3.9, range = 40–56) years at the time height and weight data was collected.We applied the same statistical analyses to decompose the twin correlations and corresponding biometric components for BMI into multiple distributions in another large independent cohort of twins for a replication study. This cohort included 13,553  twin pairs from the National Academy of Sciences-National Research Council Twin Registry (NAS-NRC Twin Registry ). Twins M = 21.9, variance = 6.0, skew = 0.98, kurtosis = 2.21) and DZ  twins. With rMZ = 0.80 and rDZ = 0.42, twin correlation is consistent with the heritability of 0.76 for BMI; a small proportion of the variance attributed to the shared family environment (2rDZ − rMZ = 0.04) and non-shared environment and measurement error (1 - rMZ = 0.20).The distribution of BMI is positively skewed for MZ (M ~ 21kg/m2) that is relatively normally distributed, and an overweight class (M ~ 24kg/m2) that is positively skewed (rMZ = 0.81 and rDZ = 0.24) are indicative of high heritability, whereas those in the overweight class (rMZ = 0.56 and rDZ = -0.05) suggest a large non-shared environmental effect and a severe violation of the classical twin model .Results showed that the two-class skew-t distribution mixture model was the best fit for the intraclass correlation model . The twoy skewed . The twiin model . Twins iThe current study presents a unique investigation of the skewed nature of BMI using latent profile analyses in a large sample of twin pairs. The BMI latent classes were estimated by including the twin correlations in the optimization process; twin pairs were therefore assigned to be in the same latent class. Our use of the skew-t distribution in the estimation accounts for the excessive skewness and kurtosis of the skewed distribution of BMI . The findings show that the skewness of the BMI distribution can be decomposed into a relatively normally distributed component within the normal BMI range, and a positively skewed component in the overweight and obese range of BMI, with the former more highly correlated in twin pairs than the latter. Twin correlations in the first class conformed to the assumptions of the classical twin model; those in the second class did not. Similar results were obtained when the analyses were replicated using another large sample of twin pairs.Our analyses suggest that two distinct processes underlie the skew of the BMI distribution and the tendency for twin correlations for BMI to violate the assumptions of the classical twin model. The first process is relatively normally distributed, in the normal range of BMI, and highly heritable. The second process is positively skewed, in the overweight and obese range of BMI, with lower twin correlations, especially for DZ twins. The near-zero DZ twin correlations represent a severe violation of the assumptions of the classical ACE twin model, and for that matter any genetically based twin model, none of which predict correlations of zero in DZ twins. In contrast to BMI, height is normally distributed, highly heritable, does not violate the classical twin model, and is well-fit by a single latent class. This contrast between height and weight is in accord with human psychological and physiological experience of height and BMI: both are under obvious genetic influence, but BMI is also subject to individual behavioral and environmental control, whereas height is not .Our finding that twins in the overweight class are more likely to be older than those in the normal class is consistent with the positive shift in BMI with age in other samples , 6. It iA few limitations of this study should be noted. First, height, weight and zygosity were all self-reported. Second, participants were primarily Caucasians, which limits generalizability of our findings to other race/ethnic groups. Future research should replicate our findings among populations of different race/ethnic composition. Third, the cross-sectional nature of our data limits our ability to examine potential differential causal relationships between BMI, genetic, behavioral, and environmental factors. Future research should make use of longitudinal studies to examine such associations, and investigate how components of BMI change or stabilize across the lifespan.Deeper understanding of the nature of these weight classes will require more information at the genetic, environmental, and phenotypic level. It would be interesting to know, for example, whether either candidate genes or polygenic risk scores are more correlated with one distribution than the other. Similarly, one could investigate how dietary or activity behaviors, or food and activity environments, differ between the classes, or how the classes change or stabilize across the lifespan.We have explored statistical models of twin development that emphasize reciprocal effects of differences in phenotype and individual behavior, demonstrating that such processes have the result of systematically depressing DZ twin correlations . To dateS1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.


To this aim, methylation studies were carried out by using genomic DNA isolated from saliva samples of 58 OSCC patients and 90 healthy controls. The frequency of the CT/AC and TT/AA genotypes was significantly higher in patients than in controls. Whereas no difference in global DNA methylation levels was observed between patients and controls, a higher frequency of methylation at both p16 and MGMT gene promoters was detected in patients compared with controls. A significant association between MTHFR gene polymorphisms and p16 and MGMT gene promoter methylation was found. The frequency of p16 and MGMT methylation was around 60% in patients with either the CT/AC or TT/AA genotype. Our results suggest that hypermethylation of cancer-related genes may be affected by MTHFR polymorphisms.Genetic polymorphisms of the methylenetetrahydrofolate reductase (MTHFR) enzyme may influence DNA methylation. Alterations in DNA methylation patterns of genes involved in the regulation of the cell cycle, DNA repair, cell adherence and metastasis process are known to contribute to cancer development. In this study, the influence of the MTHFR C677T and A1298C gene polymorphisms on global DNA methylation and site-specific methylation on  Head and neck cancer (HNC), representing the sixth most common cancer worldwide ,2, encomOral SCC (OSCC) is one of the most common types of HNC, with a considerable incidence of new cases every year. OSCC more frequently affects men than women (M:F = 2:1). The probability of developing OSCC increases with the period of exposure to risk factors, represented by a diet low in fresh fruits and vegetables, poor vitamin intake, alcohol consumption, and abuse of tobacco smoking ,4. MoreoIn recent years, hereditary factors as well as age-related mutagenic and epigenetic changes have been involved in the development of OSCC ,7.Epidemiological studies have shown that deficiency of specific nutrients, such as folate, may increase the risk of OSCC ,9. FolatMTHFR) affect the levels of available endogenous folates. MTHFR catalyzes the reduction of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, the methyl group donor for homocysteine remethylation to methionine, which in turn is converted to S-adenosyl-l-methionine (SAM), the main donor of methyl groups in different methylation reactions [MTHFR C677T polymorphism affects the enzyme’s thermal stability, resulting in the reduced activity of this enzyme [In this regard, it has been shown that gene polymorphisms of the enzyme 5,10-methylenetetrahydrofolate reductase .Genotyping of patients and controls for p = 0.00015), while the frequency of the MTHFR C1298 mutated allele was similar between cases and controls (0.23 vs. 0.28). In particular, the TT677 genotype was significantly more frequent in patients than in control subjects , while the CT677 genotype frequency only tended to be significantly higher in patients than in controls . The CC677 wild-type genotype was significantly less frequent in patients than in control subjects .The frequency of the T677 mutated allele was significantly higher in patients than in controls . The AC1298 heterozygous genotype was present in around one-third of the recruited patients and controls , while the CC1298 homozygous was mutated only in around 10% of the recruited subjects . No significant differences were found in the genotype distribution for the MTHFR genotypes in OSCC patients and healthy subjects is shown in The distribution of p > 0.05). Instead, the analysis of site-specific methylation revealed that about half of cases exhibited either a p16 or MGMT promoter region methylated with a significantly higher frequency in comparison to the controls . The promoter region of the MGMT gene was methylated in a higher number of patients in comparison with that of the p16 gene  calculation , 20 patients with the MTHFR CC/AA, CC/AC, CC/CC, and CT/AA genotypes, and in a second group, R (risk), 38 patients with the CT/AC and TT/AA genotype, which are known to be genetic determinants for alterations of MTHFR enzyme activity. We also evaluated whether the MTHFR genotype, no significant differences were observed for global DNA methylation between the two groups. The mean content of methylated cytosine was about 3.6% in both groups.After stratification of patients based on the p16 methylation was significantly higher in group R  than in group N where it was not observed (20/20 unmethylated subjects)  was higher in comparison to group N , even if this difference only tended to statistical significance   . Similar= 0.056) .Several studies suggested that aberrant methylation of DNA has an important role in the development of several cancers, such as colorectal cancer, renal cancer and esophageal squamous cell cancer ,27,28. Researchers have proved that DNA methylation is related to age, diet, and other environmental factors ,30,31. IMTHFR C677T polymorphism with the risk for the development of several human cancers has been reported [MTHFR C677T polymorphism, having major effects on MTHFR enzyme activity, was associated with an increase of oral cancer risk [The association of the reported ,35, althreported . Indeed,cer risk , while ocer risk ,39. In oMTHFR A1298C polymorphism in cancer risk is less investigated, and in certain cancers it seems to play a protective function [MTHFR genotypes were present only in 35% of controls. However, these differences were not significant. Interestingly, the frequency of the CT/AC genotype was higher than that of TT/AA (34.5% vs. 31%). The frequencies of these latter genotypes were significantly higher in patients than in controls. Noteworthy, among patients there were no subjects carrying the wild-type genotype for both polymorphisms (CC/AA). Although further studies should be carried out to better clarify the involvement of MTHFR polymorphisms, these data suggest the possible involvement of MTHFR C677T and A1298C in cancer development. The role of the function . In our To characterize the functional mechanism by which MTHFR polymorphisms may contribute to the development of OSCC in relation to the DNA methylation status, we also investigated the impact of these polymorphisms on both global and site-specific DNA methylation. Conflicting results on global DNA hypomethylation and the risk of cancer have been reported. Studies investigating DNA methylation alterations in HNC tissues showed the loss of global DNA methylation when compared to their matched normal adjacent tissues ,42,43. IInterestingly, Subbalekha and collaborators  describeUsually, LINE-1 hypomethylation measurements, which estimate a limited part of the genome, are employed to evaluate global DNA methylation. In our study, we used the capillary electrophoresis method which is able to detect the methylation of the whole genome, giving a more accurate evaluation than LINE-1 methylation. However, a limitation of this study is that the levels of cancer-specific methylation reported here may be underestimated due to the inability to distinguish the signal coming from the small percentage of tumor cells in saliva samples.p16 and MGMT promoter regions. Promoter hypermethylation in the major genes involved in cell cycle, DNA damage repair, and cancer-related signaling pathways has been extensively studied in human cancers, including ESCC [p16 negatively regulates the G1-S transition in the cell cycle and has been found frequently methylated in precursor lesions of the esophagus, and thus p16 function inactivation by hypermethylation is believed to be involved in the early stages of esophageal carcinogenesis [MGMT is the major gene in the pathway of DNA repair and has been frequently found to be silenced by CpG island hypermethylation in many cancers, including esophageal adenocarcinoma [Of note, significant differences were observed when evaluating site-specific methylation on the ing ESCC ,47,48. Aogenesis . Also thogenesis , MGMT isarcinoma .p16 and MGMT promoter methylation in patients diagnosed with OSCC than normal controls. About 50% of cases showed methylation of at least one gene and one-fifth had both p16 and MGMT gene promoters methylated. In contrast, the concomitant methylation of p16 and MGMT gene promoters was not observed in the controls. Overall, these results corroborate the concept that DNA methylation plays a major role in oral cancer development.Our study showed a higher frequency of MTHFR C677T polymorphism may influence the DNA methylation status [MTHFR TT677 genotype have a high risk of DNA hypermethylation in cancer tissues. However, only few studies carried out on a Chinese population evaluated the association of the MTHFR C677T polymorphism with aberrant CpG island hypermethylation of cancer-related genes, such as p16 and MGMT, in patients with HNC. The authors showed that the aberrant hypermethylation of p16, MGMT, and hMLH1 promoter genes was associated with the clinical characteristics of esophageal squamous cell cancer, and individuals carrying the MTHFR CT677 or TT677 genotype had a higher frequency of hypermethylation in the MGMT gene in cancer tissues [Recently, there has been growing interest to identify factors that can affect the patterns of DNA methylation. The MTHFR is an important enzyme in the one-carbon metabolism pathway that regulates the availability of methyl groups for methylation reactions, and several experimental and epidemiologic studies showed that the n status ,23,52. I tissues ,53.p16, MGMT and hMLH1, and the MTHFR C677T polymorphism in ESCC [MTHFR C677T and A1298C gene polymorphisms. Indeed, to our knowledge, this is the first study evaluating the effects of MTHFR genotypes at genetic loci C677T and A1298C on p16 and MGMT gene promoter methylation in OSCC. On the other hand, another study failed to find a relationship between aberrant DNA methylation of genes, such as  in ESCC . The disp16 gene promoter was methylated in around 60% of patients having either the CT/AC or TT/AA genotype, while it was unmethylated in patients having either the MTHFR CC/AA, or CC/AC, or CC/CC, or CT/AA genotype, and this difference was highly significant. The frequency of MGMT promoter methylation was also found to be higher, but not statistically significant, in patients with the CT/AC and TT/AA genotypes. The lack of significance for MGMT could be due to the small sample size analyzed. Interestingly, we found that the MTHFR polymorphisms may have an important role in OSCC carcinogenesis, probably due to their influence on gene-specific methylation processes.Overall, our findings suggest that the The potential mechanism by which a reduction in MTHFR activity induced by C677T and A1298C polymorphisms may affect site-specific methylation has not been investigated yet. It has been previously hypothesized that some genes could have different dosage sensitivity to alterations in methyl-donor availability in comparison to others, and the dosage sensitivity could also be dependent on the tissue type and gene-environment interactions . MTHFR polymorphisms. In addition, given the small sample size, we could not make any association with the tumor staging.However, further studies on a larger population also evaluating other possible factors acting on the one-carbon metabolism, such as the folate levels, are needed for a more extensive understanding of the regulation of the methylation process by MTHFR polymorphism and gene promoter hypermethylation, the novelty of the present study is the use of salivary samples to obtain data. In fact, based on the recently developed molecular detection methods, circulating tumor DNA can now easily be extracted from serum, plasma, saliva, broncho-alveolar lavage fluid, and urine [Even if similar previous studies have documented the association of nd urine . Moreovend urine . TherefoFifty-eight  patients with oral squamous cell carcinoma (OSCC), who referred for diagnosis to the Division of Odontostomatology at Polyclinic Hospital University of Messina, were recruited for this study. Twenty-two of them were smokers. No alcohol use was reported among patients.® DNA Self-Collection kit  from all participants, before any interventions occurred. In the same time period, 90 healthy subjects , matched with patients for age, gender, and smoking habit, were recruited on a voluntary basis among staff of Polyclinic Hospital University. Saliva samples were collected with the OrageneAll subjects gave their informed consent for inclusion before they participated in the study. The study was conducted in accordance with the Declaration of Helsinki, and the protocol was approved by the Ethics Committee of the Polyclinic University of Messina .MTHFR C677T and A1298C polymorphisms was carried out by a Real-Time PCR allelic discrimination technique, using Pre-designed TaqMan SNP Genotyping Assays .Genomic DNA was purified from saliva using Oragene DNA kit (Genotek’s), according to manufacturer’s instructions. Genotyping for For the analysis of global DNA methylation the extracted DNA was hydrolyzed by 90% formic acid. After hydrolysis, samples were evaporated and the dry residue containing free bases was dissolved in ultrapure water and immediately analyzed by capillary electrophoresis as described previously . The perp16 and MGMT genes was determined by methylation-specific PCR (MSP) after sodium bisulfite modification of DNA using a commercial kit from Sigma, according to the manufacturer’s instructions. Each MSP reaction was carried out in triplicate. The sequences of primer pairs used in MSP are shown in The methylation at the promoter region of MTHFR genotypes as well as methylation status among patients and controls were examined by using the χ2 test or Fisher’s exact test, where appropriate. A value of p < 0.05 was considered statistically significant.Statistical analysis was performed with SPSS statistical program version 13.0 . Differences in the distribution of


Background: Heart failure (HF) is a complex syndrome and is recognized as the ultimate pathway of cardiovascular disease (CVD). Studies using nutritional strategies based on dietary patterns have proved to be effective for the prevention and treatment of CVD. Although there are studies that support the protective effect of these diets, their effects on the prevention of HF are not clear yet. Methods: We searched the Medline, Embase, and Cochrane databases for studies that examined dietary patterns, such as dietary approaches to stop hypertension (DASH diet), paleolithic, vegetarian, low-carb and low-fat diets and prevention of HF. No limitations were used during the search in the databases. Results: A total of 1119 studies were identified, 14 met the inclusion criteria. Studies regarding the Mediterranean, DASH, vegetarian, and Paleolithic diets were found. The Mediterranean and DASH diets showed a protective effect on the incidence of HF and/or worsening of cardiac function parameters, with a significant difference in relation to patients who did not adhere to these dietary patterns. Conclusions: It is observed that the adoption of Mediterranean or DASH-type dietary patterns may contribute to the prevention of HF, but these results need to be analyzed with caution due to the low quality of evidence. Heart Failure (HF) is considered a complex clinical condition that compromises the heart’s ability to deliver oxygen properly to tissues. Generally, HF results from structural and/or functional dysfunction of the heart, which compromises its ability to fill itself with blood or to eject blood .Although improvements in the prevention and treatment of HF in the last decades  have beeAccording to estimates by the World Health Organization, about three-quarters of deaths from cardiovascular disease (CVD) can be prevented by measures of prevention and control of risk factors that involve changes in lifestyle . The mosSince the 1990s, with the transition from diets based on specific nutrients to an approach based on dietary patterns in CVD, promising new data emerged from randomized trials. A dietary approach to nutritional interventions in CVD had proved to be an effective strategy resulting in strong and tangible results, indicating that synergistic effects of food combined into a dietary pattern provide the maximum benefit obtainable from nutrition .The optimal dietary pattern to reduce CVD is one that emphasizes whole grains, fruits and vegetables, legumes, nuts, fish, poultry, and moderate dairy and heart-healthy vegetable oil intake; this pattern will likely reduce the CVD risk by about one-third. This healthy dietary pattern needs also to be low in refined grains, added sugars, trans fats, sugar sweetened beverages, and red and processed meats . DietaryMoreover, adherence to these healthy patterns has been shown to be associated with lower CVD risk and markers  such as Although there is evidence that supports the protective effect of certain healthy dietary patterns in CVD, to our knowledge, there is no systematic review or meta-analysis that investigates the protective effect of dietary patterns specifically on primary prevention of HF. Thus, our interest is justified to investigate systematically this effect and to assess the magnitude of the relationship between six dietary patterns and primary prevention of HF.This systematic review was conducted according to The Cochrane Collaboration  and the We found studies by searching the following data sources: Medline (accessed by Pubmed), Embase, and Cochrane Central Register of Controlled Trials database up to 20 May 2017. No filter or limitation was used during the search. References lists from selected studies were manually scanned to identify any other relevant studies.In order to obtain a high-sensitivity strategy, the following search terms were used, both as free-text and subject headings: “dietary pattern”, “diet Mediterranean”, “dash diet”, “diet vegetarian”, “diet Paleolithic”, “diet fat-restricted”, “diet, carbohydrate-restricted” and “heart-failure”.To assess comprehensively the magnitude of adherence to dietary patterns and primary prevention of HF, studies with healthy or unhealthy adult subjects were included. The studies included data comparing groups regarding adherence to dietary patterns . There was no restriction of study designs included. Experimental studies, those that included children, those that did not present dietary patterns, and those in which patients initially had HF were excluded.Titles and abstracts were evaluated independently by two reviewers (S.R.S and K.S.A). The Kappa index was calculated to assess the agreement between the two reviewers and any discrepancy was resolved by consensus or by a third reviewer (G.C.S.). The reviewers were not blind to author, institutions, or manuscript journals. Articles that did not provide sufficient information from the title and abstract were included for further evaluation, and reading was done in full. Data extraction and analysis were performed by the same two reviewers. For each study, information on publication data, population characteristics, intervention and comparison group, results, and limitations were extracted. The authors of the abstract were contacted by e-mail to provide more information about their research.Methodological quality was explored using an approach similar to that recommended by Cochrane Collaboration in risk assessment . Qualityp = 0.02). A total of 1119 studies were found, and after removal of the duplicates, 1050 were screened by title and abstract. After reading the titles and abstracts, only 14 studies were included in the systematic review . The agrThe study by Larsson et al.  was exclA total of 188.470 participants were included in this review. Four RCTs ,23,24,25Most studies included patients without previous cardiovascular disease and only two studies were performed in patients after the first infarction. There were no studies that evaluated the adherence of the low-carb and low-fat diets on the incidence of HF or worsening of the parameters of cardiac function. The follow-up period ranged from one to 21 years. Two RCTs presented a clear description of sequence generation and concealment of allocation. All studies described the losses and exclusions as well as the blindness of the outcome assessor. Regarding blinding of the researchers and participants it was considered as an uncertain risk because it is not possible to blind them. All observational studies performed the analysis describing the confounding factors. Two studies were pointed out with high risk of bias in the item on baseline differences of the participants; both were abstracts .The RCTs included in the review differed in relation to the Mediterranean diet applied to the participants.In the Papadaki et al.  and FitóIn the RCT conducted by Lorgeril et al. , no scorIn the study proposed by Tuttle et al. , the maiIn the study by Wirth et al.  and in bChrysohoou et al.  describep < 0.002). However, when assessing only the outcome of HF development, there was no significant difference (p < 0.25). Lorgeril et al. [p = 0.0001). Regarding HF outcomes, six patients in the intervention group and 11 in the control group developed HF; however, statistics of this comparison are not shown.In two randomized clinical trials, which assessed Mediterranean diet and incidence of HF, the population consisted of individuals who had already presented with their first ischemic event. Tuttle et al.  did not ce p < 0.. LorgeriA secondary arm pre-specified in PREDIMED (Prevención con Dieta Mediterránea), presented by Papadaki et al. , evaluatn = 310) or one of two traditional Mediterranean diets (TMD + virgin olive oil or TMD + nuts). After one year of intervention, both TMD decreased pro-brain natriuretic peptide (BNP), with changes reaching significance versus control group (p < 0.05). Oxidized low density lipoprotein (LDL) decreased in both TMD groups, and the decrease in the TMD + olive oil group was significantly higher than that in the control group (p = 0.004). Changes in lipoprotein (a) after TMD + olive oil were significantly lower than that in the control group (p = 0.038).During analysis of the studies, one RCT that evaluated adherence to the Mediterranean diet with HF-related markers in individuals at high cardiovascular risk was found . ParticiRegarding the cohorts that evaluated the adherence of the Mediterranean diet, three consisted of healthy individuals ,28,29 anIn the German cohort composed of 24,008 healthy middle-aged participants, it identified 209 cases of HF in 8.2 years. After multivariate adjustment, the association between adherence to the Mediterranean diet and HF was not significant, with a HR of 0.82 (95% CI 0.64 to 1.05) .p = 0.004) [In the postmenopausal women’s cohort, 1648 cases of HF were recorded, 244 of which were in the quartile with the highest adherence to the diet. The high adherence to the Mediterranean diet, compared to the groups with lower adherence, was associated with a lower risk of developing HF, with a relative risk of 0.79  .p = 0.007) and for HF, incidence was 0.69  [The male cohort, composed of 37,308 healthy individuals, identified 146 deaths from HF and 1269 cases of HF during a follow-up of 10.9 years. Adherence to the diet was inversely associated with risk of HF development, and the multivariate relative risk of HF mortality was 0.55  .p < 0.001). In addition, better adherence was associated with a lower probability of developing ventricular dysfunction after ischemic event, with an OR of 0.93 (95% 0.88–0.99), cardiac remodeling, with an OR of 0.90 (95% 0.78–1.03), and recurrent events during the two-year follow-up, with an OR of 0.88 (95% 0.80–0.98), even after adjusting for several confounding factors as well as the use of angiotensin-converting enzyme inhibitors [The fourth cohort, with 1000 individuals of both sexes with coronary artery disease, obtained a total of 459 patients who had systolic and diastolic ventricular dysfunction with a reduced ejection fraction (EF < 40%). The score on adherence to the Mediterranean diet was lower in individuals who developed ventricular dysfunction compared to that of the rest of the patients  [The elderly cohort, composed of 61% women, identified 1380 cases of HF in 21 years of follow-up. After adjusting for demographic and lifestyle variables, the DASH pattern was not associated with the incidence of HF, the highest quintile adhering to this diet presenting a relative risk of 1.05  .p < 0.001) [p = 0.006), that is a 22% lower rate of HF incidence [The cohort with postmenopausal Swedish women obtained 443 cases of HF in seven years of follow-up. Women who had a higher adherence to the DASH diet had a 37% lower rate of HF, i.e., a relative risk of 0.63  . The Swencidence .2 in systolic volume. When further adjusted for models 2  and 3 (adjusted for race/ethnicity), the associations remained significant. Regarding the ejection fraction in model 3, the increase of one point in the adherence questionnaire was associated with an increase of 0.04%, but was marginally significant (p = 0.08) [A cross-sectional study was conducted to evaluate adherence to the DASH diet and its relation to markers of cardiac function in patients from four ethnic groups without cardiovascular disease. Adherence to the DASH diet was significantly associated with diastolic volume and systolic volume when adjusted according to model 1 , smoking, high density lipoprotein (HDL), LDL, diabetes mellitus, systolic blood pressure, use of diabetes medications, antihypertensive, alcohol consumption, and physical activity). The increase of one point in the adherence questionnaire was associated with an increase of 0.31 mL in the final diastolic volume and increases of 0.12 mL/m = 0.08) .The RCT performed by Anderson et al.  investigAn abstract of a cross-sectional study by Pai et al. , who evaOur results showed that adhering to a Mediterranean or DASH type diet may be advantageous for primary prevention of HF. The results from RCTs showed a protective effect of the Mediterranean diet on the incidence of HF or worsening of cardiac function parameters for patients with previous cardiovascular disease. No studies with low-carb and low-fat diets were found.Tuttle et al.  did not In this sample of adults at high cardiovascular risk, by Papadaki et al. , the MedCohort studies also showed a clearer result on the efficacy of the Mediterranean diet in both patients with coronary artery disease (CAD) and in previously healthy patients. In addition, the three cohorts showed that the group that adhered strongly to the Mediterranean diet had a lower risk of developing HF when compared to those who adhered more superficially.Although there are differences between the studies in relation to the standard Mediterranean diet, mainly in relation to the type of addition oil which varied between olive oil and canola oil, the basis of the diet was similar when considering natural foods, whole grains, and fish consumption.The beneficial effects of olive oil have been attributed to its high content of oleic acid, a type of monounsaturated fatty acid (MUFA), as it protects against oxidative damage . In addiIt is well known that the inclusion of fish in the diet while excluding red meat, is capable of promoting benefits to human health, especially because fish is lean meat. However, the results of studies that try to point out improvements achieved by the increase in fish consumption and the attempt to correlate this consumption with the increase in omega-3 intake are still inconclusive .For the DASH diet, the results are still conflicting regarding its effectiveness for prevention of HF, even though all included studies evaluated the diet according to the same score. The cohort conducted by Del Gobbo et al.  presenteLeft ventricular (LV) function, particularly decreased end-diastolic volume, has been described as an independent risk factor for the future development of HF and cardiac death ,49,50. DThe DASH-sodium trial conducted by Sacks et al.  strengthIn addition to reducing sodium levels, the DASH dietary pattern involves increased intakes of whole grains, dietary fiber, unsaturated fatty acids, and plant proteins compared to typical Western diets . It alsoRegarding markers of cardiac function, two studies evaluated the effects of Mediterranean  and PaleOther types of diets have also been tested for primary prevention of HF. The vegetarian diet showed a positive effect on the reduction of LV diastolic dysfunction and a lower prevalence of LV hypertrophy. Finally, the RCT performed by Anderson et al.  on PaleoConsuming large amounts of vegetables and fruits, which characterizes most vegetarian diets, also brings beneficial effect to the cardiovascular system. Herbal foods provide the body with a variety of antioxidants which inhibit oxidation of LDL cholesterol, raise HDL cholesterol, and reduce total cholesterol concentrations in the circulation .It is known that chronic non-communicable diseases, such as cardiovascular diseases, are complex and multifactorial diseases and not simply developed by deficiency or excess of a certain nutrient. It is believed that regular consumers of whole foods are at a markedly reduced risk of ischemic heart disease, diabetes, and some cancers, sustaining the idea that food synergies play an important role in the prevention of chronic diseases .There is considerable evidence of food synergies and their influence on health. The synergistic effect is not surprising given the complexity of food patterns and food .We acknowledge that data on adherence to dietary patterns and the risk of developing HF are still incomplete. Due to heterogeneity of the studies, it was not possible to make the meta-analysis. Some quality parameters of the included RCTs were flawed, with omission of important information about the generation of sequence and allocation of subjects in the intervention groups. Moreover, regarding the Mediterranean diet studies, the methods used are heterogeneous with respect to the adopted pattern, with different scores being used, which can influence the adherence results. Although the dietary patterns identified may have had some common basic characteristics, each diet has been influenced by different cultural contexts.The effect of the investigated dietary patterns in the primary prevention of HF in adult individuals is not totally clear. It is possible that both DASH and Mediterranean diets may contribute to reduced incidence of HF and a decreased morbidity and mortality in adult patients, but these results need to be analyzed with caution due to the quality of evidence found. There is a need for further investigation on different dietary patterns to elucidate their relationship to primary prevention of HF.


Educational achievement was related to cardiovascular disease in some western populations, but prospective evidence on the relationship between education and the risk of incident coronary heart disease (CHD) in Asians is scarce. We aimed to explore this association and the potential modifying effect of major CHD risk factors related to education in middle-aged and older Chinese adults. We included 18,551 participants with mean age 62.8 years at baseline between September 2008 and June 2010, and followed the population until October 2013. Cox proportional hazard models and log-binomial regression models were used for multivariate analyses. Compared with participants with low education, the adjusted hazard ratio (HR) and 95% confidence interval (95% CI) for incident CHD in those with high education was 0.89 . In addition, such inverse association between education and the risk of incident CHD was more evident among individuals who were more than 60 years, physically active, overweight, and hypertension. Besides, decreases in the relative index of inequality with high education versus low education were from 0.83 to 0.76 for hypertension, from 0.85 to 0.82 for diabetes. In conclusion, education was inversely associated with the risk of incident CHD among middle-aged and older Chinese adults. Coronary heart disease (CHD) accounts for the greatest proportion of CVD, and it’s now contributing to large and rising burdens of disease mortality and disability, which increased rapidly in China2. Although some important risk factors for CHD are identified, such as overweight, obesity, hypertension, dyslipidemia, and diabetes, efforts are needed to better understand other risk factors of CHD and to develop prevention strategies.Cardiovascular disease (CVD) has become the leading cause of death worldwide3. Social inequalities in risk factors account for more than half of inequalities in CVD4. SES is usually measured by education, income, occupation, etc. The relationship between these SES measures and health outcome was the strongest for education5. As the most basic component of SES, education is prior to occupation and income and stable throughout life after young adulthood6. Low education is associated with large increases in CHD risk among men and women in high-income countries12, whereas studies on the effects of education on CHD risk in low-income and middle-income countries are more scarce13. In addition, most of these studies focused on the association between education and mortality or risk factors of CHD12. Accumulating data showed that higher education was inversely associated with the mortality or risk factors of CHD14, but only a few studies have investigated the relationship between education and the risk of incident CHD. Furthermore, education is also known to be associated with other important CHD risk factors, such as hypertension, hyperlipidemia, overweight and diabetes16. However, it remains unclear whether such risk factors would modify the association between education and the risk of incident CHD.Socioeconomic status (SES) is the root of many inequalities in healthTherefore, we aimed to (1) verify the association between education and the risk of incident CHD, and (2) investigate whether the relationship between education and the risk of incident CHD was modified by different characteristics or health status, as well as the underlying changes of CHD risk factors in China.Baseline characteristics of the 18,551 participants  are shown in Table Table P for interaction >0.05). However, the inverse association between education and risk of incident CHD was more pronounced in participants who were older than 60 years, physically active, overweight and hypertension.We further performed stratified analysis according to age, current smoking, current drinking, physical activity, overweight, hypertension, and hyperlipidemia Fig. . No signTable To the best of our knowledge, this is the first prospective study to examine the association between education and the risk of incident CHD in Chinese population. The findings from this study indicated that higher education was independently associated with a reduced risk of incident CHD, even after adjustment for a variety of potential confounders. Such relationship was more obvious among participants who were more than 60 years, physically active, overweight and hypertension. The inequalities of hypertension and diabetes increased between low and high education during the follow-up years.17, which was complex and changeable. A cohort study in Sweden reported that neighborhood socioeconomic environment was associated with the incidence of CHD18. Loucks et al. also reported a significant inverse association between life-course socioeconomic position and incidence of CHD19. Additionally, several studies examined the relationship between education and mortality from CHD7, however, the endpoint was not CHD incidence. Because of the different measures of SES and endpoint, these studies may not easily be compared. Besides, China has the examination-oriented education pattern20, socialist system, distinct cultures and values, which are different from western countries21. The upper classes in China are more likely to be married, and have better interpersonal relations, stronger power and lower stress24.Some epidemiological studies have investigated the association between SES and CHD, but the measures of SES in those studies were different, such as occupation, social support, neighborhood socioeconomic environment, and life-course socioeconomic position. For instance, the socioeconomic indicator in the Whitehall II study was occupational class9. In addition, a study from the United States indicated an inverse relationship between education and risk of CHD12. These studies supported the finding of the present study that education might be inversely associated with the risk of incident CHD. However, some potential confounders were not fully adjusted in these studies, such as diet, physical activity, stress and family history of CHD, which might mediate the relationship of education with the risk of incident CHD. In our study, we have adjusted for diet, physical activity, and family history of CHD, which did not alter the significant association between education and CHD incidence.A population-based prospective study of 5,084 participants aged between 35 and 74 reported that low education was associated with high risk of incident CHD in women after adjustment for potential confounders25, which was found to be inversely associated with all-cause mortality, especially for CHD mortality28. Secondly, strong and consistent evidence indicated that parental SES, childhood and early-life factors would contribute to the elevated risk of incident CHD30. Almost all the participants in our study were born before 1970s, when high level of education was extravagant. Thus, participants with higher education often had a higher socioeconomic gradient during their childhood, which decreased the risk of incident CHD through their lifetimes31. Thirdly, education also plays an important role in CVD by shaping employment opportunities, which are major determinants of economic resources. Individuals with high education tended to experience lower rates of unemployment, which is strongly associated with CVD32. Finally, educational inequalities also exist in financial and physical access to health care, and health-care use and quality. These factors might lead to inequalities in the diagnosis and treatment of predisposing factors of CHD, and the screening of CHD4. In the present study, use of medication for secondary prevention after hypertension, hyperlipidemia and diabetes was higher in high educational patients than low educational patients.Although the underlying mechanisms between education and CHD were unclear, several plausible mechanisms might be involved in the association. Firstly, it is widely recognized that education can lead to improved health by increasing health knowledge and healthy lifestyle behaviorset al. previously found that the inverse relationship between education and CHD was more pronounced in elderly people33, which was in line with our finding. According to the theory of cumulative advantage of education on health, education differentials in CHD incidence would be smaller at younger ages than at older ages34. Moreover, higher educational individuals with overweight or hypertension are more likely to improve their unhealthy behaviors or choose better medical care to reduce the risk of incident CHD35.In the stratified analysis, we found that the inverse association between education and the risk of incident CHD risk was more pronounced in individuals who were more than 60 years old, physically active, overweight, and hypertension, although no interaction was observed. Pekka 36. In addition, above risk factors are common predisposing factors for CHD. It is widely acknowledged that hypertension and diabetes are strong risk factors for development of CHD, and may link education to incident CHD37.Most of the previous studies on the association between education and CHD did not include the corresponding changes in CHD risk factors. In contrast, we found that high education was related to favorable changes at 5-year follow-up compared with low education in the RIIs of hypertension and diabetes. Inequalities in the prevalence of hypertension and diabetes have increased over time between low and high education. These changes might be caused by different lifestyles  and health care (usage of drug) between low and high educationThe strengths of our study are the relatively large sample size, the prospective design, validated CHD ascertainment, and the ability to adjust for a large number of potential confounders. There are also some limitations that should be taken into consideration. Firstly, we did not collect the information on income, which is also an important indicator of SES. However, education, the facet of socioeconomic status, has been reported as more determinant of health status, particularly cardiovascular conditions. It is the only indicator that remains relatively consistent through adult life. Secondly, education was only classified into two groups, including high (≥9 years) and low (≤8 years) educational level. In participants’ studenthood in China, only individuals with economic strength and strong learning ability had access to receiving high school education and above (≥9 years), which were considered as luxuries and high degrees. Thus, we select 8 years of education as the cut off.In conclusion, education was inversely associated with the risk of incident CHD in China, and this association was modified by different CHD risk factors. To our knowledge, this is the first study to investigate the association between education and the risk of incident CHD in China. Our study findings have implication for targeted preventive care of CHD. Taking targeted measures to prevent CHD for participants with low education or improving educational inequalities may reduce the disease burden caused by CHD or the health inequalities caused by social inequalities.38. In brief, a total of 27,009 retired employees of the Dongfeng Motor Corporation completed baseline questionnaires, took medical examinations, and provided fasting blood samples between September 2008 and June 2010. The follow-up survey was conducted in 2013. Among 25,978 individuals (96.2% of those at baseline) who completed the first follow-up, we excluded individuals with self-reported CHD, abnormal electrocardiogram, stroke, or cancer at baseline , as well as those with missing data on education and other covariates (n = 715), resulting in a final study sample of 18,551 individuals. Informed consent was provided by every participant. Ethics approvals were obtained from the Ethics and Human Subject committee of Tongji Medical College, Huazhong University of Science and Technology and Dongfeng General Hospital. All the methods in the present study were carried out in accordance with the approved guidelines.Data for the present analyses were based on the Dongfeng-Tongji Cohort Study, and detailed information on the design and method of this study has been reported previously39. Dietary factors included vegetable and fruit intake 40. BMI was calculated as weight in kilograms divided by height squared in meters and rounded to the nearest tenth. The individuals with BMI equal to or over 24 kg/m2 were categorized into overweight group, and others were categorized into normal group41. Hypertension was defined as a systolic blood pressure (SBP) ≥140 mm Hg, or a diastolic blood pressure (DBP) ≥90 mm Hg, or current use of antihypertensive medication, or self-reported physician diagnosis of hypertension42. High blood cholesterol was defined as total serum cholesterol (CHOL) >5.72 mmol/L or triglycerides (TG) >1.70 mmol/L or use of lipid-lowering medication, or self-reported physician diagnosis of hyperlipidemia43. Diabetes mellitus was considered as a fasting glucose (GLU) level >7.0 mmol/L or receiving anti-diabetic medications, or self-reported physician diagnosis of diabetes44.Semi-structured questionnaires were used to collected baseline data by trained interviewers. Questionnaires included information on socio-demographic characteristics , lifestyle factors , self-reported personal and family medical history, medications, stress status and occupational history. Physical activity was defined as those who took exercise more than 20 minutes per day and at least 3 times per week over half of a yearInformation on the educational attainment was assessed via questionnaire. We derived a two-group classification of education: low education and high education. Low education (8≤ years) included illiteracy, primary school and junior high school; high education (≥9 years) included high school, bachelor and master or above.46. The cases of CHD were divided into fatal CHD and non-fatal CHD according to the case definition46. We define fatal CHD events if the participant died with coronary heart disease as an underlying cause, which were ascertained by death certificates with International Classification of Diseases (ICD) codes (ICD-9 410-414 and ICD-10 I20-I25). Non-fatal events included non-fatal MI, stable or unstable angina, and coronary revascularization.All participants were covered by the Dongfeng health-care service system with specific identification number to retrieve medication information. Incident CHD cases were confirmed by review of medical insurance documentation, hospital records of the Dongfeng Central hospital and other affiliated hospitals, and death certificates. Among participants with suspicion of CHD, all the medical information were retrieved and discussed by a group of physicians in the Dongfeng Central hospital. The incidence caused by CHD included first occurrence of non-fatal myocardial infarction , fatal CHD, stable angina (SA), unstable angina (UA), and coronary revascularization, which were defined based on the diagnosis criteria of the World Health Organization by searching clinical symptoms, myocardial enzyme, ECG and coronary angiography 47. Analyses were performed using SAS version 9.4 . All statistical tests were two-tailed and considered to be significant at P value less than 0.05.Distributions of baseline characteristics of the study population were compared using One-way analysis of variance (ANOVA) for continuous variables and Chi-square tests for categorical variables. Continuous variables were described as mean ± standard deviation and categorical variables were presented as proportion. Cox proportional hazard models were used to evaluate the association between education and the risk of incident CHD calculated with HR and 95% CI, after adjustment for potential confounders. The low education was regarded as the reference group. We also evaluated the association of education with fatal and non-fatal CHD separately. We fitted three proportional hazard models. In Model 1 we only adjusted for age. Model 2 was adjusted for age, smoking status , alcohol drinking status , physical activity (yes or no), diet frequency categories (including vegetables and fruits), and stress (yes or no). Model 3 was further adjusted for BMI, waist circumference, hypertension, diabetes, hyperlipidemia, and family history of CHD. We also conducted analyses in different subgroups stratified by age, smoking status, drinking status, physical activity, overweight, hypertension and hyperlipidemia. Besides, the association between education and all-cause mortality was explored with age-adjusted cox proportional hazard models. Furthermore, age-standardized prevalence rate was used as the absolute measures, while RII was used as the relative effect size. Comparison of the degree of inequality in risk factors was carried out by calculating the RII, which is interpreted as an odds ratio, and 95% CI. The RII takes into account both the population size and the relative socioeconomic position of the groups. It was estimated by employing a log-binomial regressionSupplementary Information


Trichoderma reesei is a saprophytic fungus implicated in the degradation of polysaccharides present in the cell wall of plants. T. reesei has been recognized as the most important industrial fungus that secretes and produces cellulase enzymes that are employed in the production of second generation bioethanol. A few of the molecular mechanisms involved in the process of biomass deconstruction by T. reesei; in particular, the effect of sugar transporters and induction of xylanases and cellulases expression are yet to be known.T. reesei 69957-sugar transport system (Tr69957) is capable of transporting xylose, mannose, and cellobiose using a T. reesei 69957-sugar transport system in Saccharomyces cerevisiae. The deletion of Tr69957 in T. reesei affected the fungal growth and biomass accumulation, and the sugar uptake in the presence of mannose, cellobiose, and xylose. Molecular docking studies revealed that Tr69957 shows reduced protein–ligand binding energy for interactions towards disaccharides in comparison with monosaccharides. Furthermore, the deletion of Tr69957 affected the gene expression of cellobiohydrolases (cel7a and cel6a), β-glucosidases (cel3a and cel1a), and xylanases (xyn1 and xyn2) in the cultures of parental and mutant strains in the presence of cellobiose and sugarcane bagasse (SCB).In our study, we characterized a novel sugar transporter, which was previously identified by our group through in silico analysis of RNA-seq data. The novel Tr69957 of T. reesei can transport cellobiose, xylose, and mannose, and can affect the expression of a few genes encoding enzymes, such as cellulases and xylanases, in the presence of SCB. We showed for the first time that a filamentous fungus (T. reesei) contains a potential mannose transporter that may be involved in the degradation of cellulose.The transporter The online version of this article (10.1186/s13068-018-1084-1) contains supplementary material, which is available to authorized users. Trichoderma reesei is one of the most studied microorganisms and is known for its ability to degrade lignocellulose , which was supplemented with 5 mM uridine in the case of the pyr4 deletion strain. To perform gene expression assays, 106 cells/mL of each specific strain was inoculated into 25 mL of Mandels–Andreotti medium . The PDB file for the model was uploaded to the iGEMDOCK software along with all the ligands, and the docking was performed using standard settings, with a population size of 200 and a total of 70 generations. The post-docking parameters were analyzed and the best docked pose for each ligand was visualized by The PyMOL Molecular Graphics System, Version 1.8.6.0 Schrödinger, LLC (https://pymol.org/2/).To perform the docking analysis, structure of all the ligands was downloaded from the PubChem database . Considering all the parameters provided by the I-TASSER platform, the chosen model for the Tr69957  . Phylogenetic analysis of Tr69957 with other fungal transporters indicated that Tr69957 shows high homology with disaccharide transporters, including the maltose permeases of T. harzianum, T. gamsii, Penicillium roqueforti, and S. cerevisiae  Fig. b was conTr69957, we performed an analysis in which an S. cerevisiae strain EBY.VW4000, which lacks 20 distinct sugar transporters , cellobiose, xylose, and mannose. The drop-out assay clearly showed that the expression of Tr69957 was able to restore the growth of EBY.VW4000 on cellobiose, mannose, and xylose, suggesting that Tr69957 is able to transport these three different sugars . The consumption of cellobiose, mannose, and xylose by the S. cerevisiae EBY.VW4000 and complemented strain expressing the Tr69957 fused to the GFP was also evaluated. Under the three analyzed conditions, the yeast that does not contain the Tr69957 was unable to transport the three carbon sources after 5 days of growth.To characterize the transporter se genes ), was coTr69957 completely restored the consumption of cellobiose, xylose, and mannose after 24 h (mannose) and 36 h (cellobiose and xylose) of growth  as well as Δ69957 (6698 bp) mutant strains, respectively  cellobiose, lactose, PDA, d-(+)-raffinose, l-arabinose, raffinose, glycerol, d-xylose, or xylitol as the sole carbon-source. The Δ69957 mutant strain exhibited a decreasing radial growth in the presence of mannose, xylose, cellobiose, lactose, and fructose  with bound d-glucose, outward-occluded conformation], determined by Deng et al. [Considering all the parameters provided by the I-TASSER platform, the chosen model for the g et al. . The preTr69957 was used for the docking analysis and the results were post-analyzed based on the total free energy, as well as the Van der Waals, hydrogen, and electrostatic interactions  embedded in a lignin matrix. The hydrolysis of SCB releases a range of sugars, such as mannose, xylose, cellobiose, and fructose that can be used by the fungus for energy production. As demonstrated in Fig. 69957 mutant strain showed an impaired growth in the presence of all these sugars released from SCB hydrolysis. In this way, for the next set of experiments for characterization of Tr69957, we selected three out of the four sugars that presented significant variations during the growth conditions of the mutant strain.The early gene expression experiments performed to characterize the Tr69957 resulted in reduced fungal biomass accumulation in the presence of mannose  lacks the hexose transport activity due to the deletion of 20 hexose transporter genes and is unable to grow on glucose, fructose, mannose, or galactose as the sole carbon source. The hexose transport-deficient S. cerevisiae strain (EBY.VW4000) has been an important tool for characterizing new hexose transporters of other fungi, such as HXTB [A. niger, and the TBHXT1 [Tuber borchii. Using this S. cerevisiae strain, we can prevent interference from other sugar transporters that were already described in T. reesei. The yeast experiments suggest that Tr69957 has the capability of transporting a few sugars, including cellobiose, mannose, and xylose. Therefore, Tr69957 can transport more than one sugar, which is not surprising because the filamentous fungi are able to internalize a wide variety of mono- and oligosaccharides and it has already been shown that transporters expressed by them can often transport more than one type of sugar [For better demonstration of the phenomenon that Y.VW4000  was used as HXTB  and RhtA as HXTB  from A. e TBHXT1  transporof sugar .Tr69957 probably participated in the transport of cellobiose, mannose, xylose, lactose, galactose, and fructose in T. reesei, as △Tr69957 strain displayed growth defects on these carbon sources. However, the docking results showed that Tr69957 exhibits lower protein-binding energy in interactions involving disaccharides compared to the monosaccharides, suggesting a better orientation and affinity between these molecules.Our data showed that d-glucose can competitively inhibit d-xylose transporters as they share similarity in their structures [A. niger sugar transporters, which suggest that if the binding occurs without subsequent conformational change, then the alternative substrate will be transported across the membrane [The absence of transporter specificity can be explained by the similarity in the structures of some of the sugar molecules. Furthermore, some of these proteins can act as transporters as well as nutrient sensors . Substraructures . Howevermembrane .Conversely, the absence of specificity can be advantageous when fungi encounters complex lignocellulosic material, which contains multiple hexoses and pentoses . The funTr69957 exhibits more affinity towards cellobiose than towards mannose and xylose, the presence of these sugars can cause reduction of cellobiose uptake. Similar results were observed in MstG and MstH A. niger sugar transporters. Both transporters showed high specificity for glucose. However, some sugars like galactose, fructose, and xylose were able to inhibit MstG glucose transport by 39, 29, and 25%, respectively; in the MstH transformants, the presence of mannose and fructose reduced the uptake of labeled glucose by 70 and 50%, respectively [A. niger, substrate specificity of hexose transporters HxtB and HxtC showed a reduction in glucose uptake by 70–80% in presence of galactose, fructose, and mannose [T. reesei cellobiose transporter mutant strain. In comparison with the WT, cellobiose in the culture supernatant of STP1 mutant was consumed at a much faster rate [69957 mutant strain containing only cellobiose as the sole source of carbon  Strategy used to confirm deletion of Tr69957 by Southern blot. The figure shows the promoter region (in green) and terminator (in purple) of the gene, with the substitution of the ORF by the pyrG selection marker. A specific DNA probe was created to bind the portion of the gene (in blue), confirming its presence in the analyzed genetic content. (B) Southern blot showing the parental fungus (QM6aΔtmus53Δpyr4) used as control and four deleted candidates that did not present detection of the gene portion equivalent to 3391 bp as in the parental and presented the region corresponding to 6698bp, confirming the deletion.Additional file 4. Oligonucleotides used for evaluation of gene expression.Additional file 5. Oligonucleotides used for complementation in S. cerevisiae.Additional file 6. SCORING FUNCTIONS: the global accuracy for the five designed candidates. The chosen structure was the Model 1, which was predicted based on the 4ZWB.A (human GLUT3 glucose transporter (N45T mutant) with bound d-glucose, outward-occluded conformation), determined by Deng et al. [g et al. , since iAdditional file 7. LOCAL ACCURACY: the divergence of the modeling represented by the distance (Å) between the protein designed and the template used for the construction of it. RES: residue number; SS: predicted secondary structure ; COV: alignment coverage; BFP: predicted normalized B-factor; RSQ_1: Residue-Specific Quality of the template (the estimated deviation of the residue on the template from the built protein)).Additional file 8. INTERACTION PROFILE: the detailed amount of energy required for each atom in the protein to interact with each one of the ligands, through hydrogen bonding (H) and Van der Waals contact (V).


CO2 was reduced most significantly, decreased by 16% and 22% with compliance rates of 0.3 and 0.7, respectively.Unknown remaining time of signal phase at a signalized intersection generally results in extra accelerations and decelerations that increase variations of operating conditions and thus emissions. A cooperative vehicle-infrastructure system can reduce unnecessary speed changes by establishing communications between vehicles and the signal infrastructure. However, the environmental benefits largely depend on drivers’ compliance behaviors. To quantify the effects of drivers’ compliance rates on emissions, this study applied VISSIM 5.20  to develop a simulation model for a signalized intersection, in which light duty vehicles were equipped with a cooperative vehicle-infrastructure system. A vehicle-specific power (VSP)-based model was used to estimate emissions. Based on simulation data, the effects of different compliance rates on VSP distributions, emission factors, and total emissions were analyzed. The results show the higher compliance rate decreases the proportion of VSP bin = 0, which means that the frequencies of braking and idling were lower and light duty vehicles ran more smoothly at the intersection if more light duty vehicles complied with the cooperative vehicle-infrastructure system, and emission factors for light duty vehicles decreased significantly as the compliance rate increased. The case study shows higher total emission reductions were observed with higher compliance rate for all of CO At signalized intersections, traffic flows approach from different directions. The unknown remaining time of the signal phase generally results in extra accelerations and decelerations. These unnecessary accelerations and decelerations cause longer travel times, more complex driving modes, and increased emissions. To optimize the traffic flow and reduce harmful vehicle emissions, various cooperative vehicle-infrastructure systems (CVIS) have been developed in numerous countries. The cooperative vehicle-infrastructure system is an intelligent system that can collect the information provided by traffic infrastructure and vehicles, such as signal phases, and the vehicle’s real-time speed and position . Using t2 emissions compared with a fixed-time control scheme [In recent years, researchers have studied the environmental benefits of cooperative vehicle-infrastructure systems at signalized intersections. Bhavsar et al. studied the energy consumption reduction of plug-in hybrid electric vehicles (PHEVs) equipped with connected vehicle technology (CVT). The results indicated that the energy savings of CVT-supported PHEVs can increase by 31% to 35% when the PHEVs receive signal timing and headway data . Most stl scheme . Li et al scheme ,12.Some studies aimed at the impacts of market penetration rates of controlled vehicles. Yang et al. proposed an eco-cooperative adaptive cruise control algorithm that can compute the fuel-optimum trajectory through a signalized intersection. Simulation analyses demonstrated that for single-lane approaches, the higher the market penetration rate was, the larger fuel consumption that was saved . KatsaroIn summary, numerous studies have shown that implementing cooperative vehicle-infrastructure systems can reduce emissions and save fuel at signalized intersections. However, notably, the effectiveness of a cooperative vehicle-infrastructure system is directly affected by drivers’ compliance behaviors with the guidance information; this implies that the environmental benefits associated with the system largely depend on the drivers’ rates of compliance with the guidance information. So far, a very limited number of studies have analyzed relationships between emissions and driver compliance rates. Even some studies considered the impact from market penetration rates of vehicles controlled by cooperative vehicle-infrastructuresystems; they assumed that the driver compliance rate is 1. However, there is a difference between applying market penetration rate and drivers’ compliance rate. Market penetration rate is influenced by the development level of intelligent transportation and traffic policies. Market penetration rate cannot reflect the drivers’ different attitudes to traffic guidance when they are in different external conditions. In this context, this study strived to quantify the emissions under different driver compliance rates by using a VSP-based emission modeling approach that entailed simulating a signalized intersection with a cooperative vehicle-infrastructure system.First, in the data collection, an infrastructure data set and a traffic data set were collected at a real-world intersection through a field survey. These data were used to establish the simulation platform. A vehicle operations data set and an emission data set were selected using the MapInfo from an existing vehicle emission database for calculating emission rates.Second, in the traffic simulation, a simulation platform was developed using VISSIM. The model was calibrated and validated using the collected data. Moreover, the model was validated by comparing the observed and simulated VSP distributions. The VSP distributions were calculated using the emission model. Subsequently, a cooperative vehicle-infrastructure control strategy was implemented using the COM external program interface in VISSIM. With the control strategy, the guidance information could be provided to vehicles. Scenarios were designed and simulated. After running the traffic simulation, the simulated data under different compliance rates were obtained.Third, the emission model was developed. Based on the simulation data and the emission rates, the VSP distributions, emission factors, and total emissions were estimated for different compliance rates.Finally, a comparative analysis was conducted based on the calculation results to quantify the effects of drivers’ compliance rates on emissions at the signalized intersection equipped with the cooperative vehicle-infrastructure system.The collected data include infrastructure data, traffic data, vehicle operation data, and emission data. These data were used for the simulation platform development, model calibration, model verification, and emission rates calculation.A signalized intersection of Fuchengmen Avenue and Xisi Avenue in Beijing was selected as the test bed to develop a simulation model in this research. At this intersection, geometry, signal, and traffic flow data have been collected. Geometry data include the size of the intersection and the lanes of each approaching direction. As shown in Two types of vehicles run through the intersection: light duty vehicles and buses. Light duty vehicles are defined as cars with the gross vehicle weight ratings less than 8500 lbs. Light duty vehicles were selected for implementing the cooperative vehicle-infrastructure control. The vehicles randomly arrive at the signalized intersection. There is no platooning of vehicles with coordinated signals. The capabilities behind harmonic speed profiling with connected vehicle systems are enough.Vehicle operating data, including second-by-second speed, acceleration, and geographic information, and emission data at the study intersection, were obtained from similar intersections in the vehicle emission database established by the Transportation Environmental Lab of Beijing Jiaotong University. The influence area of the signal at the intersection was defined as a circle with a radius of 150 m. The vehicle operating data in this area were selected using the buffer function of MapInfo [Finally, a total of 40,000 second-by-second records of the vehicles’ operating and emission data at the intersection were collected, comprising 30,000 records of light duty vehicles and 10,000 records of buses. The vehicle operating and emission data were used to validate simulation model and estimate average emission rates for each VSP bin.To estimate emissions under different compliance rates, a simulation model of the intersection was developed using VISSIM. The cooperative vehicle-infrastructure system was developed and embedded in the model. The external program interface enabled adjusting drivers’ compliance rates.Providing signal status in real time;Collecting instantaneous vehicle operating data such as speed;Providing speed advice to drivers on the basis of the signal status and vehicle operating conditions.On the basis of the field data, this study developed a simulation model by using VISSIM . The coo2, NOx, HC, and CO emissions were estimated using observed and simulated data. All of the mean absolute percentage errors were found to be within 10%, signifying that the simulation model was suitable for estimating the emissions.The simulation model was calibrated using an approach based on genetic algorithms to find optimal parameter combination so that the model could accurately present the actual running states of the vehicles. Driving behavior parameters in VISSIM that are sensitive to the traffic volume and speed are calibrated, such as maximum look ahead distance, the number of observed vehicles, average standstill distance, additive part of safety distance, and waiting time before diffusion. Each parameter was divided into four levels. After using the Genetic Algorithm, the optimal parameter combination could be found. . After tThe COM interface in VISSIM was used to implement the algorithm of the cooperative vehicle-infrastructure control. In this algorithm, drivers are guided with speed advice at the intersection. Restricted by regular routes and stations, buses are not provided with speed advice. Therefore, the cooperative vehicle-infrastructure system works only for light duty vehicles. When light duty vehicles enter the influencing area (a circle of radius 150 m), their instantaneous locations and speeds are sent to the control center. According to the real-time signal status, the control center determines the guidance speed and provides advice to the vehicles . The guiTo simplify the model, the vehicle waiting time is assumed to be less than one signal cycle length at the intersection;If the effect of the leading vehicle is considered, the earliest time at which vehicle A can arrive at the stop line should be calculated using Equation (1)A can arrive at the stop line, B arrives at the stop line, and It should be noted that:A total of 11 guidance compliance rates were entered into the developed simulation model to study their effects on emissions. The minimum compliance rate is 0, which indicates that no vehicle accepts the guidance information. The maximum compliance rate is 1, which indicates that all light duty vehicles accept the guidance information. The compliance rates were set at intervals of 0.1. The market penetration rates is 1. Considering the simulation randomness and sample size, the simulation was run five times with a unique random number seed for each run under each compliance rate. Each run of the simulation simulated 1200 s of events. The output data included the vehicle type, vehicle number, speed, acceleration, and simulation time. Based on the simulation outputs, the VSP bins and vehicle emissions were estimated and analyzed.A VSP-based approach is employed to estimate emissions. The steps for emissions estimations are as follows.v is the instantaneous speed in m/s, and a is the acceleration in m/s2. For light duty vehicles, A = 0.156461, B = 0.002002, C = 0.000493, and m = 1.4788. For buses, A = 0.746718, B = 0, C = 0.002176, and m= 9.06989.As one of the parameters closely related to vehicle operating conditions, the VSP indicates the ratio of the vehicle’s output power over its mass. The VSP establishes an informative relationship between traffic emissions and driving activities, so the VSP-based model is a current trend of the emission models. MOVES, one of the VSP-based models, is a novel emissions model developed by EPA which can be used to calculate the emission factors of vehicles accurately. Based on the instantaneous speed and acceleration obtained from simulation, the VSP values for light duty vehicles and buses were calculated using Equation (2) :(2)VSP=. In this equation, the VSP data are arranged in 1 kW/ton intervals :(3)VSPbix, and CO2 for light duty vehicles and buses were obtained. Examples of emission rates of CO2 in each VSP bin for light duty vehicles and buses are illustrated in The average emission rates for each VSP bin were estimated using the vehicle operating data and emission data obtained from similar intersections in the vehicle emission database. The emission rates of CO, HC, NOi is the type of emission, i (g/km), j is the VSP bin, j for emission type i (g/s), j, T is the total travel time (s), and L is the total travel distance (km).Emission factors, which are defined as the amounts of emissions produced per kilometer, were calculated using Equation (4) .(4)EFi=i in an approaching direction of the intersection (kg); i (g/km), respectively; L is a travel distance equal to 300 m at the studied intersection.Based on the quantities of light duty vehicles and buses, the total emissions were then estimated using Equation (5):Based on the simulation platform and the emission model, the VSP distributions, emission factors, and total emissions under different compliance rates were calculated using the instantaneous speed and acceleration obtained from simulation. A comparative analysis was conducted to quantitatively evaluate the effects of drivers’ compliance behaviors on emissions at the studied intersection.VSP distributions of light duty vehicles and buses were calculated for 11 compliance rates ranging from 0 to 1 using Equations (2) and (3). The VSP distributions under compliance rates of 0, 0.3, 0.6, and 0.9 are illustrated in 2). Therefore, it can be concluded that if the compliance rate is high, then the frequencies of braking and idling are low and light duty vehicles run more smoothly at the intersection.Results showed that the VSP distributions of light duty vehicles were different for different compliance rates, as shown in Buses had no regular pattern in the VSP distributions under different compliance rates, as shown in Emission factors under different compliance rates were estimated using Equation (4), and the results are shown in 2, NOx, HC, and CO exhibited a downward trend for light duty vehicles with the increase of the compliance rate. Of the four emission types, the percentage of HC emission factor was reduced most significantly. However, the emission factors for buses did not show notable differences under different compliance rates of light duty vehicles. The fluctuation of emission factors for buses is within 3%.The results in 2, NOx, HC, and CO were 27%, 16%, 30%, and 19%, respectively. In order to analyze the variation of emission factors more clearly, the reduction percentages of the emission factors under compliance rates ranging from 0.1 to 1 were estimated. The variations for light duty vehicles were quite significant and the reduction percentages of the emission factors for buses were low . Figure 2, NOx, HC, and CO emissions. Among the four emission types, CO2 was reduced most significantly. Specifically, CO2 emissions were observed to decrease by 16% and 22% under compliance rates of 0.3 and 0.7, respectively.To demonstrate the environmental benefits more intuitively, total emissions in the southbound approach of the intersection were estimated using Equation (5). Traffic volumes were collected at the signalized intersection of Fuchengmen Avenue and Xisi Avenue through the field survey. The volumes of light duty vehicles and buses through the approach were 429 pc/h (passenger car per hour) and 9 vehicles/h, respectively. The total vehicles emissions per hour were estimated for compliance rates of 0, 0.3, and 0.7 . In addiA higher compliance rate decreases the proportion of VSP bin = 0, which means that the frequencies of braking and idling are lower, and light duty vehicles run more smoothly at the intersection if more light duty vehicles comply with the cooperative vehicle-infrastructure system.2, NOx, HC, and CO for light duty vehicles decreased significantly as the compliance rate increased. When the compliance rate was 1, the emission factors had minimums. The reduction percentages of CO2, NOx, HC, and CO were 27%, 16%, 30%, and 19%, respectively.Emission factors of CO2, NOx, HC, and CO emissions. CO2 was reduced most significantly, decreased by 16% and 21% with compliance rates of 0.3 and 0.7, respectively.Higher total emission reductions were obtained with higher compliance rate for all of COThis study investigated the effects of drivers’ compliance rates on emissions at a signalized intersection equipped with a cooperative vehicle-infrastructure system. A VISSIM-based simulation model of a signalized intersection was developed. Based on the simulation results under different compliance rates of light duty vehicles, the VSP distributions, emission factors, and total emissions were estimated. The effects of different compliance rates on emissions were analyzed by comparing the estimation results. The main findings are summarized as follows:The findings of this study provide insights for transportation practitioners to implement emission reduction policies in cities and promote environmental sustainability. In the future, the analyses could be conducted at other types of signalized intersection using different signal phases or groups of intersections with cooperative vehicle-infrastructure systems. New studies on providing guidance strategies for buses and the vehicles platoons in cooperative vehicle-infrastructure systems will be conducted.


Inocybe and Cortinarius, which form ectomycorrhizas with Dryas octopetala or the dwarf alpine Salix species. The scope of the present study was to combine Favre's historical dataset with more recent data, either with the “SwissFungi” database or with data from major studies of the French and German Alps, and with the data from novel high-throughput DNA sequencing techniques of soils from the Swiss Alps. Results of the latter application revealed, that problems associated with these new techniques are manifold and species determination remains often unclear. At this point, the fungal taxa collected by Favre and deposited as exsiccata at the “Conservatoire et Jardin Botaniques de la Ville de Genève” could be used as a reference sequence dataset for alpine fungal studies. In conclusion, it can be postulated that new improved databases are urgently necessary for the near future, particularly, with regard to investigating fungal communities from alpine regions using new techniques.Historical datasets of living communities are important because they can be used to document creeping shifts in species compositions. Such a historical data set exists for alpine fungi. From 1941 to 1953, the Swiss geologist Jules Favre visited yearly the region of the Swiss National Park and recorded the occurring fruiting bodies of fungi >1 mm  in the alpine zone. Favre can be regarded as one of the pioneers of alpine fungal ecology not least because he noted location, elevation, geology, and associated plants during his numerous excursions. However, some relevant information is only available in his unpublished field-book. Overall, Favre listed 204 fungal species in 26 sampling sites, with 46 species being previously unknown. The analysis of his data revealed that the macrofungi recorded belong to two major ecological groups, either they are symbiotrophs and live in ectomycorrhizal associations with alpine plant hosts, or they are saprotrophs and decompose plant litter and soil organic matter. The most frequent fungi were members of  The Swiss geologist Jules Favre was one of the first mycologists who explored the macrofungal communities in the alpine zones. Employed as a curator at the Natural History Museum of Geneva, he visited the region of the Swiss National Park (SNP) every summer between the years 1941 and 1957 making daily excursions to sample and record the fruiting bodies of fungi larger than 1 mm and easily visible to the naked eye . He published the list of fungi occurring in the alpine zone (above timberline) in 1955 Favre, . HoweverDryas octopetala, Salix spp., Kobresia myosuroides, and Bistorta vivipara have been conducted in alpine regions of the Alps and Scandinavia. Investigations followed either a classical way by collecting fruiting bodies .The main source of information was the book of Favre . Becausehttp://www.nationalpark.ch) was founded in 1914 and, is the oldest National Park of the Alps. It is located in the southeast of Switzerland and has 170 km2 of natural landscape including subalpine forests, alpine grassland, and nival peaks up to 3,174 m of height. The SNP is situated on a mosaic of various bedrocks, but dominated by dolomitic limestone and only occasionally interrupted by various other limestone formations. Some parts consist of acidic bedrock formed by granite gneiss of crystalline base or of sandstones, shales, and psephites of the Permian . What Favre (https://map.geo.admin.ch).The SNP . According to that “Index,” many genus names have changed since Favre's descriptions, e.g., Rhodophyllus changed to Entoloma, Geophila to Psilocybe, and the genus Hygrophorus has been splitt into several genera, e.g., Hygrophorus s.s., Hygrocybe, Gliophorus, or Cuphophyllus  and some major studies from the Swiss  or from the village of S-charl (during 7 years) Figure . His earFrom S-charl most of the excursions lead Favre into the nearby valleys Val S-charl 16x) and Val Sesvenna (13x)  and Murtaröl (11), and association with the acidophilic S. herbacea in Ils Lajets (17) The genera of fungi with the highest number of recorded species were by Favre  were Inos Tables , 4. A ras Tables . The higLichenomphalina spp. Other lichenized fungi were not recorded, although known to be abundant in the alpine zone. Overall, 95 ectomycorrhizal, 105 saprotrophic, and 4 lichenised taxa were recorded, resulting in a total of 204 taxa , and some were recorded only once.Overall, Favre  recordedts Table , whereasDryas as well as with all of the four dwarf willows .From the whole range of the ECM species found by Favre  22% werews Table . Thirty ls Table . About ols Table  , and Bistorta vivipara (Inocybe dulcamara). However, only B. vivipara is known to host ECM fungi, whereas J. communis hosts preferentially arbuscular mycorrhizal (AM) fungi and L. procumbens ericoid fungi  (Tables Inocybe (20 newly described taxa) and Cortinarius (20), followed by Entoloma (3), and Hebeloma, Arrhenia, and Clitocybe (each 2). With so many type specimens assigned, all sampling sites of Favre  Tables , S2. Theof Favre   have been included recently in mycological surveys and phylogenetic analyses by Cripps et al. . A smaller portion is only associated with the four Salix species . Dominant genera are Inocybe with 99 species (49%) and Cortinarius with 54 species (26%). Besides the “SwissFungi” database, which included 130 species, the compilation of the French Alps from Kühner and Lamoure . About half of these fungi are generalists  and associated with es Table . A small Lamoure  contribuSalix herbacea and S. helvetica, but also Alnus viridis and Pinus mugo at the lower margin of the site. Investigation of the soils resulted in 2,390 fungal operational taxonomic units  in total. More than two thirds of the OTUs were assigned to the Ascomycetes, about 25% to the Basidiomycetes, and 5% remained unclassified. In the Basidiomycetes, a large group were Agaricales, with 120 OTUs, whereas Russulales, Boletales, and Cantharellales were less abundant. The majority of the classified Agaricales found were saprotrophic and not typical alpine species . The genera Cortinarius, Inocybe, and Entoloma, with 5 to 6 OTUs each, were not particularly abundant, and some of their identified members were not typically known as alpine species .The forefield of the “Damma” glacier  or with Salix herbacea . Based on the DNA identification of ECM root tips from the three ECM plants D. octopetala, Salix polaris and Bistorta vivipara in Svalbard (Norway), Botnen et al.  associated with n et al.  came to Notwithstanding the extraordinary pioneering achievement of Favre , his worCortinarius aureomarginatus , C. croceus, and C. diasemospermus , INSDC , or UNITE . Thus, we propose an enhanced effort to sequence type material from fungal collections according to accepted standards, which include reliable sequence data combined with a correct taxonomic name as well as collection and voucher information (Schoch et al., Overall, we are convinced that there is an urgent need to improve databases with sequences from historic type material. As stated by Nilsson et al. , preciseThe book of Favre  represenIB, FG, and BS analyzed the data of Favre . FG and 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.