Risk assessment of venous thromboembolism in inflammatory bowel disease by inherited risk in a population-based incident cohort

Inflammatory bowel disease(IBD),including Crohn’s disease(CD)and ulcerative colitis(UC),is a chronic inflammatory disease of the digestive tract with increasing prevalence globally.Although venous thromboembolism(VTE)is a major complication in IBD patients,it is often underappreciated with limited tools for risk stratification.AIM To estimate the proportion of VTE among IBD patients and assess genetic risk factors(monogenic and polygenic)for VTE.METHODS Incident VTE was followed for 8465 IBD patients in the UK Biobank(UKB).The associations of VTE with F5 factor V leiden(FVL)mutation,F2 G20210A prothrombin gene mutation(PGM),and polygenic score(PGS003332)were tested using Cox hazards regression analysis,adjusting for age at IBD diagnosis,gender,and genetic background(top 10 principal components).The performance of genetic risk factors for discriminating VTE diagnosis was estimated using the area under the receiver operating characteristic curve(AUC).RESULTS The overall proportion of incident VTE was 4.70%in IBD patients and was similar for CD(4.46%),UC(4.49%),and unclassified(6.42%),and comparable to that of cancer patients(4.66%)who are well-known at increased risk for VTE.Mutation carriers of F5/F2 had a significantly increased risk for VTE compared to non-mutation carriers,hazard ratio(HR)was 1.94,95%confidence interval(CI):1.42-2.65.In contrast,patients with the top PGS decile had a considerably higher risk for VTE compared to those with intermediate scores(middle 8 deciles),HR was 2.06(95%CI:1.57-2.71).The AUC for differentiating VTE diagnosis was 0.64(95%CI:0.61-0.67),0.68(95%CI:0.66-0.71),and 0.69(95%CI:0.66-0.71),respectively,for F5/F2 mutation carriers,PGS,and combined.CONCLUSION Similar to cancer patients,VTE complications are common in IBD patients.PGS provides more informative risk information than F5/F2 mutations(FVL and PGM)for personalized thromboprophylaxis.

Genetic susceptibility loci of lung cancer are associated with malignant risk of pulmonary nodules and improve malignancy diagnosis based on CEA levels

Objective:The heightened prevalence of pulmonary nodules(PN)has escalated its significance as a public health concern.While the precise identification of high-risk PN carriers for malignancy remains an ongoing challenge,genetic variants hold potentials as determinants of disease susceptibility that can aid in diagnosis.Yet,current understanding of the genetic loci associated with malignant PN(MPN)risk is limited.Methods:A frequency-matched case-control study was performed,comprising 247 MPN cases and 412 benign NP(BNP)controls.We genotyped 11 established susceptibility loci for lung cancer in a Chinese cohort.Loci associated with MPN risk were utilized to compute a polygenic risk score(PRS).This PRS was subsequently incorporated into the diagnostic evaluation of MPNs,with emphasis on serum tumor biomarkers.Results:Loci rs10429489G>A,rs17038564A>G,and rs12265047A>G were identified as being associated with an increased risk of MPNs.The PRS,formulated from the cumulative risk effects of these loci,correlated with the malignant risk of PNs in a dose-dependent fashion.A high PRS was found to amplify the MPN risk by 156%in comparison to a low PRS[odds ratio(OR)=2.56,95%confidence interval(95%CI),1.40−4.67].Notably,the PRS was observed to enhance the diagnostic accuracy of serum carcinoembryonic antigen(CEA)in distinguishing MPNs from BPNs,with diagnostic values rising from 0.716 to 0.861 across low-to high-PRS categories.Further bioinformatics investigations pinpointed rs10429489G>A as an expression quantitative trait locus.Conclusions:Loci rs10429489G>A,rs17038564A>G,and rs12265047A>G contribute to MPN risk and augment the diagnostic precision for MPNs based on serum CEA concentrations.

Evaluating the potential of(epi)genotype‑by‑low pass nanopore sequencing in dairy cattle:a study on direct genomic value and methylation analysis

Background Genotype-by-sequencing has been proposed as an alternative to SNP genotyping arrays in genomic selection to obtain a high density of markers along the genome.It requires a low sequencing depth to be cost effective,which may increase the error at the genotype assigment.Third generation nanopore sequencing technology offers low cost sequencing and the possibility to detect genome methylation,which provides added value to genotype-by-sequencing.The aim of this study was to evaluate the performance of genotype-by-low pass nanopore sequencing for estimating the direct genomic value in dairy cattle,and the possibility to obtain methylation marks simultaneously.Results Latest nanopore chemistry(LSK14 and Q20)achieved a modal base calling accuracy of 99.55%,whereas previous kit(LSK109)achieved slightly lower accuracy(99.1%).The direct genomic value accuracy from genotype-by-low pass sequencing ranged between 0.79 and 0.99,depending on the trait(milk,fat or protein yield),with a sequencing depth as low as 2×and using the latest chemistry(LSK114).Lower sequencing depth led to biased estimates,yet with high rank correlations.The LSK109 and Q20 achieved lower accuracies(0.57-0.93).More than one million high reliable methylated sites were obtained,even at low sequencing depth,located mainly in distal intergenic(87%)and promoter(5%)regions.Conclusions This study showed that the latest nanopore technology in useful in a LowPass sequencing framework to estimate direct genomic values with high reliability.It may provide advantages in populations with no available SNP chip,or when a large density of markers with a wide range of allele frequencies is needed.In addition,low pass sequencing provided nucleotide methylation status of>1 million nucleotides at≥10×,which is an added value for epigenetic studies.

A novel multimodal prediction model based on DNA methylation biomarkers and low-dose computed tomography images for identifying early-stage lung cancer

Objective:DNA methylation alterations are early events in carcinogenesis and immune signalling in lung cancer.This study aimed to develop a model based on short stature homeobox 2 gene (SHOX2)/prostaglandin E receptor 4gene (PTGER4) DNA methylation in plasma,appearance subtype of pulmonary nodules (PNs) and low-dose computed tomography (LDCT) images to distinguish early-stage lung cancers.Methods:We developed a multimodal prediction model with a training set of 257 individuals.The performance of the multimodal prediction model was further validated in an independent validation set of 42 subjects.In addition,we explored the association between SHOX2/PTGER4 DNA methylation and driver gene mutations in lung cancer based on data from The Cancer Genome Atlas (TCGA) portal.Results:There were significant differences between the early-stage lung cancers and benign groups in the methylation levels.The area under a receiver operator characteristic curve (AUC) of SHOX2 in patients with solid nodules,mixed ground-glass opacity nodules and pure ground-glass opacity nodules were 0.693,0.497 and 0.864,respectively,while the AUCs of PTGER4 were 0.559,0.739 and 0.619,respectively.With the highest AUC of0.894,the novel multimodal prediction model outperformed the Mayo Clinic model (0.519) and LDCT-based deep learning model (0.842) in the independent validation set.Database analysis demonstrated that patients with SHOX2/PTGER4 DNA hypermethylation were enriched in TP53 mutations.Conclusions:The present multimodal prediction model could more efficiently distinguish early-stage lung cancer from benign PNs.A prognostic index based on DNA methylation and lung cancer driver gene alterations may separate the patients into groups with good or poor prognosis.

Impact of cognition-related single nucleotide polymorphisms on brain imaging phenotype in Parkinson’s disease

Multiple single nucleotide polymorphisms may contribute to cognitive decline in Parkinson’s disease. However, the mechanism by which these single nucleotide polymorphisms modify brain imaging phenotype remains unclear. The aim of this study was to investigate the potential effects of multiple single nucleotide polymorphisms on brain imaging phenotype in Parkinson’s disease. Forty-eight Parkinson’s disease patients and 39 matched healthy controls underwent genotyping and 7 T magnetic resonance imaging. A cognitive-weighted polygenic risk score model was designed, in which the effect sizes were determined individually for 36 single nucleotide polymorphisms. The correlations between polygenic risk score, neuroimaging features, and clinical data were analyzed. Furthermore, individual single nucleotide polymorphism analysis was performed to explore the main effects of genotypes and their interactive effects with Parkinson’s disease diagnosis. We found that, in Parkinson’s disease, the polygenic risk score was correlated with the neural activity of the hippocampus, parahippocampus, and fusiform gyrus, and with hippocampal-prefrontal and fusiform-temporal connectivity, as well as with gray matter alterations in the orbitofrontal cortex. In addition, we found that single nucleotide polymorphisms in α-synuclein(SNCA) were associated with white matter microstructural changes in the superior corona radiata, corpus callosum, and external capsule. A single nucleotide polymorphism in catechol-O-methyltransferase was associated with the neural activities of the lingual, fusiform, and occipital gyri, which are involved in visual cognitive dysfunction. Furthermore, DRD3 was associated with frontal and temporal lobe function and structure. In conclusion, imaging genetics is useful for providing a better understanding of the genetic pathways involved in the pathophysiologic processes underlying Parkinson’s disease. This study provides evidence of an association between genetic factors, cognitive functions, and multi-modality neuroimaging biomarkers in Parkinson’s disease.

Evaluation of polygenic risk score for risk prediction of gastric cancer

Genetic variations are associated with individual susceptibility to gastric cancer.Recently,polygenic risk score(PRS)models have been established based on genetic variants to predict the risk of gastric cancer.To assess the accuracy of current PRS models in the risk prediction,a systematic review was conducted.A total of eight eligible studies consisted of 544842 participants were included for evaluation of the performance of PRS models.The overall accuracy was moderate with Area under the curve values ranging from 0.5600 to 0.7823.Incorporation of epidemiological factors or Helicobacter pylori(H.pylori)status increased the accuracy for risk prediction,while selection of single nucleotide polymorphism(SNP)and number of SNPs appeared to have little impact on the model performance.To further improve the accuracy of PRS models for risk prediction of gastric cancer,we summarized the association between gastric cancer risk and H.pylori genomic variations,cancer associated bacteria members in the gastric microbiome,discussed the potentials for performance improvement of PRS models with these microbial factors.Future studies on comprehensive PRS models established with human SNPs,epidemiological factors and microbial factors are indicated.

Genome-wide associations, polygenic risk, and Mendelian randomization reveal limited interactions between John Henryism and cynicism

BACKGROUND John Henryism(JH)is a strategy for dealing with chronic psychological stress characterized by high levels of physical effort and work.Cynicism is a belief that people are motivated primarily by self-interest.High scores on the JH scale and cynicism measures correlate with an increased risk of cardiovascular disease.High cynicism is also a hallmark of burnout syndrome,another known risk factor for heart disease.AIM To evaluate possible interactions between JH and cynicism hoping to clarify risk factors of burnout.METHODS We analyzed genetic and psychological data available from the Database of Genotypes and Phenotypes for genome-wide associations with these traits.We split the total available samples and used plink to perform the association studies on the discovery set(n=1852,80%)and tested for replication using the validation set(n=465).We used scikit-learn to perform supervised machine learning for developing genetic risk algorithms.RESULTS We identified 2,727,and 204 genetic associations for scores on the JH,cynicism and cynical distrust(CD)scales,respectively.We also found 173 associations with high cynicism,109 with high CD,but no associations with high JH.We also produced polygenic classifiers for high cynicism using machine learning with areas under the receiver operator characteristics curve greater than 0.7.CONCLUSION We found significant genetic components to these traits but no evidence of an interaction.Therefore,while there may be a genetic risk,JH is not likely a burnout risk factor.

Study on Inheritance Mechanism of Tomato Fruit Firmness

[Objective] This study aimed to investigate the inheritance mechanism of tomato fruit firmness.[Method] Two tomato cultivars significantly different in fruit firmness were selected for investigation of the inheritance mechanism of tomato fruit firmness using combination analysis of six generations （P1,P2,F1,F2,B1 and B2）.[Result] The results indicated that the heredity of tomato fruit firmness was consistent with the additive-dominant model controlled by one pair of major genes; the additive effect （d）,dominant effect （h） and degree of dominance （h/d） of major genes were 17.37,-7.96 and-0.46,respectively,showing positive additive effect and incompletely dominant-negative effect; the hereditability of major gene effect in B1,B2 and F2 generation was 88.59％,45.81％ and 85.62％,respectively.[Conclusion] The heredity of fruit firmness was controlled by one pair of major gene,showing significant additive effect and dominant effect.

β－苯乙胺N－取代的反应中重排反应的机理研究

以ＰＯＬＹＧＥＮ软件中的ＣＨＡＲＭｍ程序和集团坐标轮换法，对６个化合物的结构进行计算机分子模拟，根据所得化合物的能量最低构象，计算了键合能、电子总能量、偶极矩、胺基氮原子电荷以及氧原子电荷等值并作为结构参数对反应的反应热进行估算，同时对β－苯乙胺Ｎ－取代反应中的重排反应（反应式（１））进行机理研究，结果认为重排产物Ｎ－（２－苯乙基）Ｎ－（２－羟乙基）－对硝基苯胺４是由Ｎ－（２－苯乙基）－２－（对硝基苯氧基）乙胺３经分子内亲核取代反应所得，此反应可能为Ｓｍｉｌｅｓ重排反应。

Genetic Analysis on Plant Height in Rice in Different Growing Seasons

[Objective] The aim was to carry out the genetic analysis on plant height of rice（Oryza sativa L.）cultivated in different seasons.[Method] Three rice parents with great difference in plant height including CB1（83.1 cm）,CB4（105.5 cm）and CB7（115.6 cm）were chosen to construct two parental combinations：CB1×CB4 and CB7×CB4,and the corresponding filial generations P1,F1,P2,B1,B2 and F2 were obtained.The 6 populations were planted in middle and late seasons respectively to measure their height traits.The Akaike＇s information criterion（AIC）of the mixed major gene and polygene model was used to indentify the existence of major genes affecting quantitative traits in B1,B2,F2 populations.When the major genes existed,the genetic effects of the major genes and polygenes and their genetic variance were estimated through segregation analysis.[Result] One additive major gene plus additive-dominance polygenes was the most fitted genetic model for the trait in all B1,B2,F2 populations in two planting seasons.The heritability values of the major genes varied from 38.63% to 78.53% and those of polygenes varied from 1.72% to 36.04%,and the total heritability values were 45.52-92.93%.The additive effect d value of the two genetic populations under two planting seasons was-4.56,-9.16,-7.19,and-9.38,respectively,as suggested that additive effect of the major genes would decrease the express of the plant height trait.[Conclusion] The heritability of plant height trait was affected by planting seasons and the combinations clearly as a whole.

Genetic Analysis of Embryo Production Frequency in Wheat × Maize Cross

A DH population derived from C49S-87/01Y1-1069 was used to study the inheritance of wheat haploid embryo production frequency（EPF） in wheat × maize cross with the mixed major gene and polygene inheritance model of quantitative traits. The results showed that the EPF of wheat × maize cross was controlled by two dominant epistatic genes and polygene with gene effects of 1.95 for the first major gene, 6.69 for the second one and 2.80 for the polygene. The inheritability of major genes was as high as 72.09%, suggesting that the differences in EPF among wheat materials were mainly influenced by genotype. However, non-genetic factors were still important, especially for wheat materials with low EPF.

2,4-二芳基-1,3-二硫戊环化合物的定量构效关系研究

目的 为进一步设计和研究 2 ,4-二芳基 - 1,3 -二硫戊环类化合物提供可参考依据。方法 对合成的 68个 2 ,4-二芳基 - 1,3 -二硫戊环化合物以 POL YGEN软件中的 CHARMm程序对其活性化合物的结构进行计算机分子模拟。根据所得化合物的能量最低构象 ,计算了其 VDW体积、偶极矩、总键能、总键角能、总非正则能以及硫原子电荷等值 ,将其结果作为结构参数分别与这些化合物的 i NOS抑制活性进行相关分析。结果 以逐步回归法建立了相关性方程 :- log IC50 =2 .2 847+ 0 .10 5 2 X3+ 0 .0 5 89X6 - 0 .0 0 44 X1 8( n=16,R=0 .919466,S=0 .7112 2 5 ,F=2 1.8760 5 )。结论 二硫戊环类化合物的 i NOS抑制活性与分子总键角能、分子总范德华能呈正相关 ,与二硫戊环 2位芳基相连

Analysis of Flesh Thickness of Summer Squash by Using Mixed Major Genes and Polygene Inheritance Model

[Objective]The aim was to study the inheritance of flesh thickness,so as to provide a theoretical basis for breeding of thicker-flesh summer squash variety.[Method]Using mixed major genes and polygene inheritance model,a joint analysis of six generations from two crosses q-1×23-4G（Cross 1）and q-1×A-7（Cross 2）was conducted to investigate the inheritance of the flesh thickness of summer squash.[Result]The results showed that the flesh thickness was controlled by D-2 model（a pair of additive major genes plus additive-dominant polygene）;genetic effect of flesh thickness in the two crosses was mainly the dominant effect of polygene.Heritability values of the genes of F2 generation were relatively low,while the environmental effect was great.[Conclusion]To flesh thickness,the method of individual selection on high separated generations was effective in the breeding of summer squash with thick flesh.

Analysis of Fruit Length Using Mixed Major Gene and Polygene Inheritance Model in Summer Squash

[Objective]The aim was to provide scientific basis for breeding on the fruit length of summer squash.[Method]Inheritance of the fruit length of summer squash in P1,P2,F1,B1,B2 and F2 from two crosses q-1 ×23-4G（cross 1） and q-1 ×A-7（cross 2） was investigated by the mixed major gene and polygene inheritance model of quantitative traits.[Result]The results showed that the fruit length was controlled by D2 model;Genetic effect of fruit length of cross 1 was mainly additive effect,but genetic effect of fruit length of cross 2 was mainly dominant effect.Heritability values of the genes of F2 were relatively high and the environmental effect was relatively low.[Conclusion]The fruit length breeding of summer squash should be selected in early generation selection.

Analysis of Fruit Width Using Mixed Major Gene and Polygene Inheritance Model in Summer Squash

[Objective] The aim was to provide scientific basis for breeding on the fruit width of summer squash.[Method] Inheritance of the fruit width traits of summer squash in P1,P2,F1,BC1,BC2 and F2 from two crosses q-1×23-4G（cross 1）and q-1×A-7（cross 2）was investigated by the mixed major gene and polygene inheritance model of quantitative traits.[Result] The fruit width was controlled by B-1 model.Heritability values of the major genes of F2 were relatively high and the environmental effect was relatively low.[Conclusion] The fruit width breeding of summer squash should be selected in early generation selection.

Genetic Analysis of Cryotolerance in Cotton During the Overwintering Period Using Mixed Model of Major Gene and Polygene

The joint analysis of the mixed genetic model of major gene and polygene was conducted to study the inheritance of cryotolerance in cotton during the overwintering period.H077（G.hirsutum L.,weak cryotolerance） and H113（G.barbadence L.,strong cryotolerance） were used as parents.Cryotolerance of six generation populations including P1,P2,F1,B1,B2,and F2,from each of the two reciprocal crosses H077×H113 and H113×H077 were all investigated.The results showed that cryotolerance in cotton during the overwintering period was accorded with two additive major genes and additivedominance polygene genetic model.For cross H077×H113,the heritabilities of major genes in B1,B2,and F2 were 83.62,76.84,and 90.56%,respectively;and the heritability of polygene could only be detected in B2,which was 7.76%.For cross H113×H077,the heritabilities of major genes in B1,B2,and F2 were 67.42,68.95,and 83.40%,respectively;and the heritability of polygene was only detected in F2,which was 6.51%.In addition,the whole heritability in F2 was always higher than that in B1 and B2 in each cross.Therefore,for the cryotolerance breeding of perennial cotton,the method of single cross recombination or single backcross should be adopted to transfer major genes,and the selection in F2 would be more efficient than that in other generations.

Interaction of major genes predisposing to hepatocellular carcinoma with genes encoding signal transduction pathways influences tumor phenotype and prognosis

Studies on rodents and humans demonstrate an inherited predisposition to hepatocellular carcinoma （HCC）. Analysis of the molecular alterations involved in the acquisition of a phenotype resistant or susceptible to hepatocarcinogenesis showed a deregulation of G1 and S phases in HCC of genetically susceptible F344 rats and a G1-S block in lesions of resistant Brown norway （BN） rats. Unrestrained extracellular signal-regulated kinase （ERK） activity linked to proteasomal degradation of dual-specificity phosphatase 1 （DUSP1）, a specific ERK inhibitor, by the CKS1-SKP2 ubiquitin ligase complex occurs in more aggressive HCC of F344 rats and humans. This mechanism is less active in HCC of BN rats and human HCC with better prognosis. Upregulation of iNos cross-talk with IKK/NF-KB and RAS/ERK pathways occurs in rodent liver lesions at higher levels in the most aggressive models represented by HCC of F344 rats and c-Myc-TGF-α transgenic mice. iNOS, IKK/NF-κB, and RAS/ERK upregulation is highest in human HCC with a poorer prognosis and positively correlates with tumor proliferation, genomic instability and microvascularization, and negatively with apoptosis. Thus, cell cycle regulation and the activity of signal transduction pathways seem to be modulated by HCC modifier genes, and differences in their efficiency influence the susceptibility to hepatocarcinogenesis and probably the prognosis of human HCC.

Genetic Analysis of Stripe Disease Resistance in Rice Restorer Line C224 Using Major Gene plus Polygene Mixed Effect Model

The inheritance of stripe disease resistance in a rice restorer line C224 was analyzed using the mixed effect model of major gene plus polygene for quantitative traits.In addition,the resistance was investigated in seven crosses of C224 with maintainer lines.The results showed that the stripe resistance of C224 was controlled by two major genes with additive-dominance-epistasis effects plus polygenes with additive-dominance effects （E-1 model）.These two genes had additive effects of-12.47% and-24.75%,respectively,showing negative dominance effects.There were significant epistasis and interaction effects between the two major genes.The heritability of the two major genes was 92.12%,while that of polygenes was 2.74%,indicating that the stripe resistance had dominant major gene effect.Of the seven crosses,five displayed high or medium resistance to the stripe disease.

The power of a healthy lifestyle for cancer prevention:the example of colorectal cancer

Objective:We aimed to directly compare the estimated effects of adherence to a healthy lifestyle with those of risk predisposition according to known genetic variants affecting colorectal cancer(CRC)risk,to support effective risk communication for cancer prevention.Methods:A healthy lifestyle score(HLS)was derived from 5 lifestyle factors:smoking,alcohol consumption,diet,physical activity,and body adiposity.The association of lifestyle and polygenic risk score(PRS)(based on 140 CRC-associated risk loci)with CRC risk was assessed with multiple logistic regression and compared through the genetic risk equivalent(GRE),a novel approach providing an estimate of the effects of adherence to a healthy lifestyle in terms of percentile differences in PRS.Results:A higher HLS was associated with lower CRC risk(4,844 cases,3,964 controls).Those adhering to all 5 healthy lifestyle factors had a 62%(95%CI 54%-68%)lower CRC risk than those adhering to≤2 healthy lifestyle factors.The estimated effect of adherence to all 5 compared with≤2 healthy lifestyle factors was as strong as the effect of having a 79 percentile(GRE 79,95%CI 61-97)lower PRS.The association between a healthy lifestyle and CRC risk was independent of PRS level but was particularly pronounced among those with a family history of CRC in≥1 first-degree relative(P-interaction=0.0013).Conclusions:A healthy lifestyle was strongly inversely associated with CRC risk.The large GRE indicated that CRC risk determined by polygenic risk may be offset to a substantial extent by adherence to a healthy lifestyle.

Genetic segregation analysis of unsaturated fatty acids content in the filial generations of high-linolenic-acid rapeseed(Brassica napus)

Alpha linolenic acid(ALA)is an essential polyunsaturated fatty acids that can improve human health.Rapeseed is the second largest oil crop in the world but the ALA content in its seed fatty acids is only about 10%.Two rapeseed germplasms YH25005 and R8Q10 with high content(up to 21%)ALA were developed by intervarietal crossing.They were used as the maternal parent(P_(1))when crossed with a low ALA parent SW(P2)to produce the seeds of the F1 hybrid,F2,and backcrosses to P1(BCP1)and P2(BCP2).A multigeneration joint segregation analysis was conducted to determine major gene t polygene effects of the content of three major unsaturated fatty acids including oleic(OA),linoleic(LA)and ALA.The results showed that,although some genes favorable to ALA accumulation were not allelic in R8Q10 and YH25005,all the inheritances of OA,LA and ALA in YH25005×SW and R8Q10×SW followed a genetic model of‘two pairs of additive major genes t additive-dominant polygenes’.It is suggested that the contents of OA,LA and ALA are closely related and the major genes in the parent SW containing loss-of-function mutations in FAD2 and FAD3 loci had strong effect to reduce ALA and elevate OA.However,total genetic effect of the polygenes was greater than that of the two major genes,especial on ALA content.It indicated that it is necessary to employ a larger F2 population to find the plant that accumulate enough minor-effect polygenes for high ALA content.The results are useful for high ALA rapeseed breeding and future work of gene mapping.