Olea europaea L. subsp. europaea var. europaea), the most important oil tree crop of the Mediterranean basin, has been poorly evaluated up to now. A deep sequence analysis of fragments of four genes, OeACP1, OeACP2, OeLUS and OeSUT1, in 90 cultivars, revealed a wide range of polymorphisms along all recognized allele forms and unexpected allele frequencies and genotype combinations. High linkage values among most polymorphisms were recorded within each gene fragment. The great sequence variability corresponded to a low number of alleles and, surprisingly, to a small fraction of genotype combinations. The distribution, frequency, and combination of the different alleles at each locus is possibly due to natural and human pressures, such as selection, ancestrality, or fitness. Phylogenetic analyses of allele sequences showed distant and complex patterns of relationships among cultivated olives, intermixed with other related forms, highlighting an evolutionary connection between olive cultivars and the O. europaea subspecies cuspidata and cerasiformis. This study demonstrates how a detailed and complete sequence analysis of a few gene portions and a thorough genotyping on a representative set of cultivars can clarify important issues related to sequence polymorphisms, reconstructing the phylogeny of alleles, as well as the genotype combinations. The identification of regions representing blocks of recombination could reveal polymorphisms that represent putatively functional markers. Indeed, specific mutations found on the analyzed OeACP1 and OeACP2 fragments seem to be correlated to the fruit weight.Gene sequence variation in cultivated olive ( Olea europaea L. subsp. europaea var. europaea) is the most important oil tree crop of the Mediterranean basin. The species is characterized by an unusually rich germplasm, represented by a high number of traditional varieties, vegetatively propagated over 1000s of years , followed by empirical local selection, probably associated to episodic varietal substitution, leading to the current varietal framework  genes (SUT1) (LUS)  synthesis and desaturation, as well as in other metabolic pathways  genes , a gene s (SUT1)  and anot1) (LUS)  were chopathways . The funmulation , meanwhiic acids . The SUThe plant . Differehe plant . In olivcterized . The LUS cyclase , is invo cyclase . Up to n cyclase .Polymorphic fragments of these genes were deeply analyzed within a large set of olive varieties, with the aims to: (i) understand the level of genomic variation within the chosen fragments; (ii) assign the alleles to each olive varieties; (iii) understand evolutionary and phylogenetic relationships among haplotypes, and (iv) verify potential correlations between detected polymorphisms and phenotypic traits.O. europaea subsp. cuspidata from Nepal and subsp. cerasiformis from the Canary Islands were used to clarify the ancestry and the phylogenetic relationships of the cultivar allele groups, whereas one sample of the related species Olea paniculata, belonging to the subgenus Paniculatae .In order to obtain polymorphic E. coli XL1 blue strain. DNA from 10 colonies for each genotype was amplified using the ExTaq (Takara) following the manufacturer instructions for the reaction and PCR amplification. Direct sequencing in both directions of the PCR products was performed on an ABI 3130 Genetic Analyzer  using the ABI Prism BigDye Terminator v.3.1 Ready Reaction Cycle Sequencing Kit (Applied Biosystems). To evaluate the presence of polymorphisms, sequences were aligned by BioEdit 7.1.7 (BioEdit1) and by Geneious 6.1.6 (Geneious2). Afterward, new allele-specific primers were designed and applied to amplify distinct alleles at each locus in all 90 cultivars . Amplifications were performed on a thermal cycler PCR System 9600 , using the following cycling conditions: initial denaturation at 95°C for 5 min, followed by 50 cycles of 95°C for 30 s, 59°C for 30 s and 72°C for 90 s, with a final elongation at 72°C for 10 min. Amplicons of different lengths were obtained. Considering that different fragment lengths prevent direct sequencing, in order to identify and distinguish the two alleles at each of the 20 diploid genotypes, amplicons of each locus were cloned, by using pGEM-T Easy Vector (Promega) and In order to verify whether the obtained alleles effectively belong to a single locus, a co-segregation analysis was performed by genotyping 94 individuals of a F1 intervarietal cross progeny (Leccino × Dolce Agogia), through length and SNP markers at each locus heterozygous in at least one parental variety (data not shown).O. europaea subspecies cuspidata and cerasiformis and the related species O. paniculata.The same cloning strategy was also applied to obtain at least one allele for each sample of the 34O. europaea subsp. europaea var. sylvestris5,6, in order to find the position of these genes on the chromosome.The obtained sequences were used as queries in a BLAST search on the genome of the cultivars Farga 3 and LeccAt each locus, the number of SNPs, indels, SSRs and repeats was calculated. The frequency of polymorphisms was obtained by the ratio between number of polymorphisms and total fragment length. Linkage analysis among polymorphisms was evaluated through MapMaker/EXP Version 3.0b .2 parameters and computing a non-linear regression line. The 180 fragments sequenced for each gene were analyzed, as well as the haplotypes inferred by the software. Due to the nature of the analyzed fragments, in this manuscript the terms ‘allele’ and ‘haplotype’ have been used as synonymous. The same software was used to detect the degree of intragenic recombination (IR) in real and inferred haplotypes at the four loci using genotypic data with the r2 parameter. The same software was used for haplotype reconstruction which was implemented by PHASE with 1,000 iterations, thinning intervals equal to 10 and 1,000 burn-in iterations, and either forcing or not the program to use the naive Gibbs sampler (GS options)  among polymorphisms at each locus. The degree of LD was estimated by calculating roptions) . The twoO. paniculata sample, used as an out-group, and one allele for each of the two subspecies cuspidata and cerasiformis.Median joining phylogenetic networks based on SNP and length markers for each allele were obtained by Network 4.6.11 software , includiOeACP1–OeACP2 for 56 genotypes, were analyzed by following the same parameters through the same software.The evolutionary history was inferred using the Neighbor-Joining method by MEGA7 software . The optTo assess whether signatures of positive selection or demographic processes were present in each of the four candidate loci, three neutrality tests were separately performed   on exonsOeACP genes, a subset of 56 varieties from the Olive Varietal Collection established in Zagaria  was phenotyped for FA composition and fruit weight. Fruit samples were collected from two plants per cultivar, for 2 years and two dates/year. Methods of analysis refer to 7.Following the neutrality test results, to verify the presence of any genotype/phenotype correlation for Furthermore, Box and Whisker plots were chosen to graphically show the correlations between genotype and phenotypic traits . For eacOeACP1 and OeACP2 genes, separately. This software uses a Bayesian model that accounts for the linkage present between polymorphic sites within aligned sequences. For each gene, BAPS carried out a genetic mixture analysis to determine the best partition of the data into K groups. The mixture analysis was run with 100 iterations, the input number of reference genotypes from each group was 10; the input number of iterations for reference genotype was 100, the minimum size of a group was 10. Subsequently, BAPS carried out a genetic admixture analysis to estimate admixture proportions within each genotype, taking into consideration the most likely number of K groups identified previously. TASSEL genotype alignments were first filtered using minimum counts of 56 sequences, with a minimum SNP frequency of 0.05 , including all sites for each fragment.Finally, TASSEL was used to establish genotype/phenotype relationships by using the association analyses with general linear model (GLM) and mixed lineage model (MLM), the latter only for significant correlations by using kinship matrix obtained by the same software. A Q matrix was built by using the Bayesian model-based approach and the “Clustering with linked loci” module, as implemented in BAPS 6.0 , inferriOeSUT1, and eight SSRs, the latter exclusively present in the OeACP2 fragment.Combined approaches, including cloning and direct sequencing performed by means of locus- and allele-specific primers, allowed us to distinguish all alleles at the four loci on 90 olive cultivars. Nucleotide diversity in the gene fragments is summarized in Table OeACP gene fragments, as detailed below.Single nucleotide polymorphism frequency was 7.9 times higher than length polymorphisms . The variation density was one polymorphism every 19.5 bp (only one every 21.9 bp for SNPs), one every 45.9 bp on exons and one every 17 bp on introns. The 17 SNPs detected in coding regions included 11 transitions and six transversions, leading to amino acid changes only in the OeACP1 allele fragments, with a maximum length of 990 bp, revealed 29 SNPs and four indels, giving a frequency of one polymorphism every 30 bp. A high degree of linkage was observed: on the 33 polymorphisms, only two SNPs at positions 407 and 503 and one indel at position 537 were unlinked, whereas the remaining (90.9%) were grouped in three linkage groups.The analysis of the OeACP2 allele fragments, with a maximum length of 1,363 bp, showed higher variability than that found in the other loci. The 127 polymorphisms included 112 SNPs, seven indels and eight SSRs, with a frequency of one polymorphism every 10.7 bp. The OeACP2 fragments carried insertions and various microsatellite regions, made up of mono-, di-, and tri-nucleotide motifs. As for OeACP1, a high linkage degree was observed: 104 polymorphisms (81.9%) were linked in five linkage groups.The In this gene fragment (555 bp), ranging from the seventh exon to a portion of the ninth exon, only 10 nucleotide mutations were detected, with a frequency of one every 55.5 bp. Four SNPs were unlinked, whereas the remaining were grouped into two linkage groups.OeSUT1 allele fragments (maximum length of 914 bp) revealed 23 SNPs, two indels of 1 and 19 bp, respectively, and a motif of 166 bp that in some cultivars was repeated twice in a head-tail arrangement, generating a tandem repeated sequence. The alignment of the isolated gene fragments with the SUT1 sequence of other plant species – i.e., Populus trichocarpa (HM749897.1), Spinacia oleracea (X67125.1), Vitis vinifera (HQ323258.1), or Verbascum phoeniceum (FJ797307.1) – showed that the first exon of the olive gene was divided into two portions by a tandem repeated sequence made up of a 166 bp motif. By aligning this sequence with the collection of repetitive DNA elements of Dfam database (Dfam8), it was not identified as a part of a transposable element. The frequency of polymorphisms was one every 35.2 bp. Fifty percent of polymorphisms were linked into two linkage groups. No linked SNPs were found among the four gene fragments.Overall, the sequence analysis of the OeACP1, 12.2% in OeACP2, 38.9% in OeLUS, and 24.4% in OeSUT1). The co-segregation analysis of alleles confirmed that all four genes effectively belong to four distinct loci (data not shown).The list of alleles found at each locus, along with the corresponding GenBank accession numbers, is reported in Supplementary Table OeACP1. The allele named 1A was clearly distinct from the others, due to two deletions at positions 306 and 802  and a 12 bp insertion at position 485, joined with 23 linked SNPs. Three out of the four indels were linked. The other alleles belonged to the same group called 3 , and were distinguished among each other by a few polymorphisms. Three out of four polymorphisms found in exon two generated three amino acid changes  were found in our sample set. Out of these, the most frequent genotype was 2D-4A (18.9%), derived from the combination of the most frequent alleles , and only 12 of the possible 28 genotypes were observed, the most common being C–C, shared by the 35% of cultivars Figure .OeSUT1 alleles were divided into three groups: long (L), medium (M), and small (S). Length differences between S and M alleles were due to a 19 bp indel, whereas L alleles contained a duplication of a 166 bp motif in respect to M alleles. SNPs in the exon sequences did not produce any amino acid change. The M group represented 60% of total alleles and, within this group, M2 showed the highest frequency. The S alleles represented 27% of the total, and the two L alleles only the remaining 13%. No random combinations of OeSUT1 alleles were observed, in fact, only 22 out of the possible 120 (18.3%) genotypes were identified. The most frequent genotypes were M2-S4 and M2-M2. Taken together, they represented more than 44% of the observed ones  , each representing a ‘haplotype block.’ In OeACP2, the IR for calculated haplotypes was 0.252, while the IR obtained with real haplotypes was 0.023, corresponding to a calculated minimum number of 19 recombination events versus the true five ones. The latter were located at 140–147, 147–151, 318–348, 348–470, and 851–1,167 bp. Interestingly, one of them included the last part of the second exon (from 318 to 337 bp). All the other OeACP2 portions represented haplotype blocks. In OeSUT1, the IR value was four times higher in calculated versus true haplotypes, and the only RE was located at 259–566 bp, in the intron region containing the 166 bp repeated motif , whose separation from the other alleles of group 2 was sustained by a bootstrap value of 100% , linking the sequences at each locus, are shown in Figure OeACP1-1A, OeACP2-4A, OeLUS-C, OeSUT1-M2)  and for OeACP2 SNP 869  with fruit weight  percentage  and OeACP2 (p = 0.047) .When the functional contribution of polymorphisms at each OeACP1 and OeACP2 sequences of 56 cultivars, in order to verify if the cultivars that grouped in the same genetic cluster showed also a relationship with the fruit fresh weight. Almost all cultivars carrying both or only one of the two alleles OeACP1-3D and OeACP2-4F are included in three clusters  and showed a medium-high fruit weight  and single nucleotide polymorphisms (one every 21.9 bp) was high compared to that found in other plant species, in which SNP frequency was detected by the same sequencing method. The values detected in olive were higher than that observed not only in perennial woody species, such as  species , but witultivars . Interesultivars . This diultivars .The frequency of indels and microsatellites along the introns of the analyzed fragments of the four loci resulted quite low compared to total polymorphisms . Due to the high SNP frequency and the low indel frequency, their ratio resulted higher than in other plant species: 12.4 against 6.4 in apple , 3.6 in OeSUT1 gene sequences respect to other species. The first exon was interrupted by an intron, not recognized as a part of a transposable element . Several hypotheses can be conceived to explain this evidence: (i) a different level of ancestrality may have affected the chance of combination; (ii) the complex varietal diversification process  could haO. paniculata species, was probably due to the existence of unsampled or extinct ancestral sequences. The high affinity between the allele clusters showed that the different allele forms derived from each other through single or multiple nucleotide mutations. Moreover, the relationships established in our material may be considered as an evidence of introgression of alleles in cultivated olives from other subspecies. Interesting is the case of OeACP1-1A allele, highly widespread among cultivars of different geographical origin, that appeared before all other alleles, including those carried by different subspecies, and for which a different introgression process could be hypothesized. In OeACP2, a close affinity with subsp. cuspidata can be deduced for alleles of group 4, whereas group 2 alleles derived from subsp. cerasiformis. A similar frame can be seen in OeLUS, in which the two subspecies are distinctly linked to different allele groups. It is interesting to highlight that cerasiformis subspecies is characteristic to the extreme western side of Mediterranean, while cuspidata subspecies spreads from north-east and south of Africa to the south of Asia, two completely different ways of diffusion.The use of single-copy genes to reconstruct phylogenies of the olive complex, has been limited by the high allele diversity, coupled to numerous ancient admixture events, because very different topologies may be supported by different loci . In thisAs interpreted for the distribution of lupeol synthase alleles in wild Mediterranean olives , the recOeACP2-2L, which is more present in the cultivars of the West-Central Mediterranean and characterizes a large number of Spanish cultivars. The same result was obtained by specific Neighbor-Joining analysis.This also explains why in the set of cultivars analyzed, which included the main commercial ones, it was not possible to group the varieties according to their geographical origin. Despite this, some alleles or allelic associations have been reasonably regionalized, see the French cultivars and the case of OeACP2) were specific to single varieties , thus representing powerful markers, exploitable not only for genotyping purposes but also for olive oil DNA testing, parentage analysis and association mapping studies.Due to the strong association, only 21% on total polymorphisms resulted informative, but these were able to discriminate a vast majority of the analyzed cultivars. Some polymorphisms , and other six double-purpose (oil and table olives) varieties . Box plot graphs highlighted also three SNPs in OeACP2 sequences as significantly related to the percentage of palmitoleic acid. When each relationship was tested by TASSEL, only the correlation between fruit weight and both SNPs in 4F and 3D alleles remained within the significance range, when using GLM analysis. Box and Whisker plots, produced on the basis of the allele estimates output provided by TASSEL, underlined that when the estimate values were >0.5 the effect of allele’s genotypic class clearly emerged. In particular, homozygous genotypes were associated with lower phenotypic values.This low recombination level, together with a very high and significant values of Tajima test, encouraged us to investigate the occurrence of any genotypic/phenotypic correlation. Box and Whisker plots showed a good correlation between fruit weight and two specific polymorphisms, the ACP genes are characterized, in the upstream and downstream sequences, by regulatory sequences that could play a major role in affecting the trait and, in an oil crop as olive, genes involved in oil accumulation may have undergone to a strong selective pressure, shaping the distribution, frequency and combination of the different alleles. Further functional genomic studies are needed to validate these SNPs as effective putative causal polymorphisms.The lack of strong marker/phenotype correlation may be due to numerous factors. Fruit weight and FAs content are quantitative traits presumably controlled by multiple loci and the effect of one over the others is still totally unknown. Acyl carrier protein is a cofactor and a structural protein that serves for the plastidial condensation of the acyl chains C16, but it represents a member of the FAs synthesis chain that involves different actors . ACP genThis research showed how an in-depth gene sequence analysis may help to clarify important issues related to allele variation and apportioning of variability in a perennial cultivated species.The polymorphisms found could be applied for the discrimination and evaluation of the olive genetic resources, in studies on the evolution and domestication of the crop, or as a starting point for further functional genomic experiments.NC, LB, MC, RM, and VS conceived and designed the experiments. NC, VS, FA, RM, SM, and CR performed the experiments. RM, LL, NC, LB, and FA analyzed the data. LB and MC contributed reagents, materials, and analysis tools. LB, NC, RM, LL, SM, FA, and MC wrote the manuscript. All the authors read and approved the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Traumatic head injuries occur frequently in Taiwan, having catastrophic consequences for the victims, their families, and society as a whole. However, little is known about the risk factors at the population level in Taiwan. The primary aim of this study was to obtain more information on these variables and their relationships. Another aim was to analyze the effects of independent variables such as sex, age, residency, pre-existing conditions, mechanisms of injury, associated injuries, and severity on the probability of in-hospital death. Using the 2007–2008 total admissions claim dataset from Taiwan’s National Health Insurance system, total admissions due to acute head injury were selected for further analysis. The obtained data included patient demographics and trauma hospitalization rate. A total of 99,391 patients were admitted with head injury, 48,792 of which had moderate-to-severe head injury. There were 4935 cases recorded as in-hospital mortality and the standardized in-hospital mortality rate was 10.7 deaths per 100,000 person-years. The mortality rate increased with age. After adjustments, male sex, age older than 54 years, living in a rural area, lower monthly income, a Charlson comorbidity index greater than one, being a pedestrian hit by a motor vehicle, fall from a height, and having significant chest, abdominal, or lower extremity injury increased the risk of death during admission. This population-based analysis provides information about the incidence rate and death rate for admissions in Taiwan due to acute head injury and the factors that affect in-hospital mortality. Our results that highlight the risk factors for adverse outcome can help us prevent or improve rural area trauma care of head injury patients in the future. Trauma is a devastating cause of morbidity and mortality in Taiwan and other countries around the world. Head injury is still one of the leading causes of death in modern society ,2,3. AppIn Taiwan, the National Health Insurance (NHI) system covers nearly 99% of the population and this study retrieved the entire claim admissions dataset for those who were coded with discharged diagnostic numbers defined by the International Classification of Diseases, 9th revision, Clinical Modification (ICD-9-CM). Therefore, this study included all NHI trauma patients who were hospitalized and discharged between 1 January 2007 and 31 December 2008 and is population-based. We captured all the trauma hospitalizations of patients defined as any admission claim coded with at least one ICD-9-CM diagnosis between 800.00 and 959.99, except 905.00 to 909.99, which indicate “late effects of injuries, poisonings, toxic effects and other external causes: 930.00 to 939.99, which include “effects of a foreign body entering through an orifice”, and 958, “complications”. In Taiwan, computed tomography (CT) for patients with traumatic head injury is generally covered by NHI and moderate-to-severe head injury was mainly diagnosed by radiological findings on CT, as in other studies . This re2) is higher in urban areas (2635 persons/km2) than in rural areas (230 persons/km2). According to the level of monthly income, injured patients were classified into four groups: monthly income less than USD $660, between USD $660 and $1320, more than USD $1320, and dependents, which are those who were injured but not employed, including those insured (NHI) by a spouse, other family member, or social welfare. During the study period, the average Taiwan monthly living cost was USD $595/person. Pre-existing conditions were defined as the medical co-morbidities of the injured patients who were diagnosed and recorded with ICD-9-CM codes in the same admission data file [. We used Charlson’s comorbidity index to quantify the PECs [According to capacity and services, hospitals in Taiwan were classified into three levels: medical centers (MC), regional hospitals (RH), and local hospitals (LH). Taiwan’s definition for medical centers and regional hospitals includes capacity and services. Empirically, the patients were stratified into nine age groups: 0–14, 15–24, 25–34, 35–44, 45–54, 55–64, 65–74, 75–84, and >84 years of age. The patient area of residency was classified into two groups: urban (including suburban) and rural areas. The average population density  and severe head injury , as shown in ICD E-codes in the claim data were also used to classify the mechanism of injury . A patiep < 0.05. All the data analyses were generated by using SPSS software, version 20.0 . Institutional board review was waived because this study involved secondary data analysis of a well-encrypted database.The percentage of sex, four categories of monthly income, four categories of PECs, and mortality rates were compared to understand the differences between all patients with head injuries and the patients with moderate-to-severe head injuries. Means, medians, and standard deviations of continuous variables, such as age, monthly income, Charlson comorbidity index, and ICD/ISS were analyzed using analysis of variance (ANOVA). The relative odds of death in the admission course were estimated by using binary logistic regression with the backward conditional stepwise method. Statistical significance was set at During 2007–2008, 476,241 cases were retrieved as trauma admissions. There were 50,599 cases of mild head injury in which the AIS in the head region was 1 or 2. Among these cases, 151 deaths occurred and the in-hospital mortality rate was 0.3%. There were 9666 cases of moderate head injury (AIS head of 3) and 126 deaths were recognized; the in-hospital mortality rate was 1.3%. There were 39,126 cases of severe head injury  and 4809 deaths were recognized; the in-hospital mortality rate was 12.3%. The raw hospitalization rate for all head injuries in Taiwan was 215.3 per 100,000, and for moderate-to-severe head injury, 105.9 per 100,000. After calculations using the U.S. 2000 standard population, the standardized incidence of all head injury was 220.6 per 100,000 in Taiwan and the standardized incidence of moderate-to-severe head injury was 110.5 per 100,000. p < 0.01). Among the patients who had in-hospital mortality, 2038 were registered in rural areas. The rural population yielded a higher raw hospitalization death rate for patients with moderate-to-severe head injury than the urban population . Monthly distribution of case numbers during 2007–2008 is shown in In Taiwan, 6,935,205 people resided in rural areas in 2008. During the study period, 41,570 rural people were admitted with head injury and 18,108 of these rural people had moderate-to severe head injury. Thus, focusing on moderate-to-severe head injury, the patients who resided in rural areas had a higher hospitalization rate than those residing in urban areas .We included characteristics of the patients and their injuries, including age, sex, rural residency, level of monthly income, Charlson comorbidity index, mechanism of injury, and ICD/ISS in a logistic regression to estimate the adjusted relative odds of in-hospital fatality for the patients with moderate-to-severe head injury. The results are shown in As an indicator of PECs, higher Charlson comorbidity index scores were associated with a higher risk of death. The relative odds of dying in the group with a Charlson comorbidity index of two was 1.52 times higher than those for the group with a Charlson comorbidity index of zero (95% CI: 1.34–1.72) and the relative odds of dying in the group with a Charlson comorbidity index more than two was 2.5 times higher (95% CI: 2.16–2.90). Several mechanisms of injury did not significantly affect the probability of death. Compared with falls on the same level, only pedestrians hit by a motor vehicle and falls from a height had a higher probability of in-hospital death. Furthermore, being associated with another significant injury significantly increased the risk of mortality; the mortality rate in the group associated with a significant abdominal, chest, or lower extremity injury was 3.41, 3.41, and 1.37 times higher, respectively, than the group without a significant injury.Head injury continues to be a major health problem around the world in both developed and developing countries ,2,3,4. TAmong those aged 15 to 74 years, the most common mechanism of injury is motorcycle-related traffic accident because motorcycles are the most popular transportation vehicles in Taiwan. Motorcycle riders and passengers should be the target population for injury prevention, even in the age group between 65 to 74 years. For those older than 65 years, falls caused more than two-thirds of the injuries and hospitalization rates in this age group. This deserves more attention because trauma does not only affect the younger population; therefore, a focus on injury prevention programs should be aggressively advocated for the elderly. For the people younger than 15 years old in Taiwan in the study by Tsai et al. in 2004, the incidence rate was higher in the age groups of 4 to 9 and 10 to 14 years. The main cause of pediatric head injury was traffic injury followed by falls. Of all pediatric traffic injuries, motorcycle-related injury had the highest incidence, followed by pedestrian and bicycle-related injury .In the United States, head injuries cause about 2.5 million emergency department (ED) visits, 280,000 hospitalizations, and 50,000 deaths annually . In TaiwOur regression model showed that only pedestrians hit by a motor vehicle and those who had a fall from a height were associated with a higher probability of mortality; other causes of head injury were not. This result might be caused by the lack of helmet protection and pedestrians hit by a motor vehicle is a well-known dangerous mechanism of injury . In TaiwThis study had some limitations. First, this is a retrospective study including all the NHI claim data about injury-related hospitalizations during the study period. However, the accuracy of NHI claim data is assured given the severe penalty for fraud and erroneous claiming. Second, this dataset does not include the Glasgow Coma Scale (GCS), which may be misleading for assessing the severity of head injury. The NHI covers brain CT for head injury in an emergency setting, and the severity score ICD/ISS in this study was basically determined by brain CT. The third limitation is that nearly 10% of all admissions were recorded as traffic accidents without further information about the mechanisms, and one-quarter of admissions were recorded without any information about the mechanism of injury. This is reasonable because there was a higher proportion of trauma patients who were found injured without any witnesses or other information source compared to in the United States . BecauseIn conclusion, this epidemiological study that focused on traumatic head injury hospitalizations in Taiwan is the first population-based study providing abundant objective information about the severity of the damage caused by head injury in our society. Although the government and medical institutes have attempted to mitigate the damage caused by head injury for decades, head injury still requires public attention to alleviate the impact on human health and society. Our results that highlight the risk factors for adverse outcome can help us to prevent or improve rural area trauma care for head injury patients in the future.


An intracellular ion channel may have a central role in the release of cytokines by macrophages. Related research article Plesch E, Chen C-C, Butz E, Scotto Rosato A, Krogsaeter EK, Yinan H, Bartel K, Keller M, Robaa D, Teupser D, Holdt LM, Vollmar AM, Sippl W, Puertollano R, Medina D, Biel M, Wahl-Schott C, Bracher F, Grimm C. 2018. Selective agonist of TRPML2 reveals direct role in chemokine release from innate immune cells. eLife7:e39720. doi: 10.7554/eLife.39720When a pathogen enters our body, innate immune cells called macrophages respond by making and releasing cytokines, small proteins which recruit other agents of the immune system to help combat the infection . Many ofIon channels are gate-like proteins that control many biological processes by enabling ions to pass through membranes. For example, a channel called TRPML2 belongs to a family that allows the movement of positive ions, such as calcium ions . This grIn contrast, TRPML2 is less well studied and characterized at a molecular level. It is mostly found in immune cells, and the expression of TRPML2 in a macrophage increases when the cell encounters lipopolysaccharides. In addition, macrophages from mice in which the gene for TRPML2 is deactivated fail to secrete CCL2 in response to lipopolysaccharides . Now, inPlesch and collaborators went through a library of small molecules to identify several agonists of TRPML2, testing each candidate on human embryonic kidney cells that express this channel in the plasma membrane. These molecules were designed so they could also reach the ion channels that were present deep within the cell, in the endosomes and lysosomes. An increase in calcium ions inside the cell meant that a small molecule was ‘turning on’ the channel. The compounds that showed promising effects were then more rigorously tested on a new set up which involved tearing open a single cell with a pipet, and then using a second pipet to record ion currents in experimentally enlarged lysosomes and endosomes, thanks to a method known as patch clamping . Thus, tBy treating cells with ML2-SA1, the researchers – who are based in Italy, Germany and the United States – show that when the TRPML2 channels are open, macrophages can secrete CCL2 and recruit more cells. TRPML2 is inactive at acidic pH, so it is unlikely that CCL2 is transported in structures called lysosomes, which are profoundly acidic: indeed, the cells did not release any lysosomal proteins. Early and late endosomes, on the other hand, are mildly acidic and express TRPML2, which makes them more probable vehicles for CCL2. However, carriers known as Golgi vesicles cannot be excluded .When TRPML2 opens, positive ions, and in particular calcium ions, can move across the membrane; the resulting changes in ion composition inside and outside the endosomes may promote all aspects of CCL2 trafficking, involving both fission and fusion of vesicles and the secretion of CCL2 from the cell itself .Having a selective TRPML2 agonist will help to find molecules that decrease the activity of the channel. These drugs could then be used to treat immune diseases that are associated with an enhanced release of CCL2, such as psoriasis, multiple sclerosis, rheumatoid arthritis and atherosclerosis .


How frequently and when do suicidal ideation, plan, and attempt emerge in adolescence and young adulthood?In a cross-sectional, epidemiological study of 1180 adolescents and young adults from a random community sample in Germany, 10.7%, 5.0%, and 3.4% of participants reported lifetime suicidal ideation, plan, and attempt, respectively. The cumulative incidence of suicidal behavior increased after age 10 years more strongly among female than male participants, and the transition from ideation to action occurred mostly during the same year.Adolescence and young adulthood are critical developmental periods for the onset of suicidal behavior, with a short time for targeted interventions to disrupt the ideation-to-action pathway. This cross-sectional epidemiological study of a random community sample of adolescents and young adults in Dresden, Germany, examines the prevalence and timing of suicidal ideation, planning, and attempt. Suicidal behavior is a leading cause of death among adolescents and young adults. In light of the ideation-to-action framework, the delineation of frequency and temporal characteristics of such behavior during this developmental period is crucial.To provide lifetime and 12-month prevalence estimates of suicidal behavior, including ideation, plan, and attempt, in adolescents and young adults of the general population, and to provide information about age at onset, temporal characteristics of suicidal behavior, including duration (number of years between onset and last occurrence) and frequency (number of episodes), and transition patterns across suicidal behaviors.A cross-sectional epidemiological study was conducted in a random community sample of 1180 adolescents and young adults aged 14 to 21 years assessed in 2015 to 2016 in Dresden, Germany. Data analysis was performed from October 2018 to March 2019.Lifetime and 12-month suicidal behavior  were assessed with a standardized diagnostic interview  by trained clinical interviewers. The onset, frequency, and duration of suicidal behavior were assessed by questionnaire.P = .04), for plan , and, among those older than 14 years, for attempt . Of those with suicidal ideation, 66.0% reported persistent or recurrent ideation over more than 1 year with 75.0% reporting more than 1 episode. Of the participants with lifetime suicidal ideation, 47.0% reported a suicide plan and 23.9% reported a suicide attempt. The transition to suicide plan or attempt occurred mainly in the year of onset of suicidal ideation or plan; of those who transitioned, 74.9% transitioned from ideation to plan, 71.2% transitioned from ideation to attempt, and 85.4% transitioned from plan to attempt in the same year.Of the 1180 participants , 130 participants , 65 participants , and 41 participants  reported lifetime suicidal ideation, plan, and attempt, respectively. Any lifetime suicidal behavior was reported by 138 participants . Age-specific cumulative incidence estimates indicated an increase in suicidal behavior during adolescence, starting at age 10 years (<1%), increasing slightly until the age of 12 years (2.2%), and then increasing sharply thereafter until age 20 years (13.5%). There were different patterns among female and male participants for ideation, plan, and attempt, with an overall higher incidence among female participants for ideation (hazard ratio, 1.51; 95% CI, 1.02-2.22; There is an urgent public health need for timely identification of suicidal behavior in adolescents and young adults to terminate persistent or recurrent suicidal tendencies and to interrupt the ideation-to-action transition. In addition to the suggested use of advanced multivariate approaches to improve prediction ,17 a thorough descriptive temporal characterization of suicidal behavior is necessary to improve the identification of critical pathways.Previous research18 cross-national research in adult samples illustrated that approximately one-third of those with suicide ideation continue to make a plan, and less than 30% make an attempt.3 The transition from ideation to plan or attempt occurs often during the first year of onset of ideation.19 There is mixed evidence on whether the age at onset and duration or persistence of ideation or plan increases the risk for transition to more severe types of suicidal behavior.24Considering the ideation-to-action transition,25 have examined the temporal characteristics of suicidal ideation, plan, and attempt, such as onset, duration, frequency, and transition patterns, in adolescence and young adulthood. To our knowledge, only 1 study4 has examined the distribution of age at onset and speed of transition in adolescents, pointing to very limited information about the pathways of suicidal behavior in this critical age group. Therefore, the goal of the present study is to provide (1) lifetime and 12-month prevalence estimates of suicidal behavior, including ideation, plan, and attempt; (2) information about age at onset; (3) temporal characteristics of suicidal behavior, including duration and frequency; and (4) information about transition across suicidal behaviors among adolescents and young adults from a regional general population sample in Germany.Only a few epidemiological studies26 a community-based sample of 1180 adolescents and young adults aged 14 to 21 years from Dresden, Germany, was assessed via face-to-face standardized diagnostic interview by trained clinical interviewers. Participants received €50 as an incentive. Detailed information on sampling and study procedures can be found elsewhere.26In the cross-sectional epidemiological Behavior and Mind Health (BeMIND) study,STROBE) reporting guideline.The BeMIND study was performed in accordance with the ethical standards of the 1964 Declaration of Helsinki and its later amendments, and the study protocol was approved by the ethics committee of the Technische Universität Dresden, Germany. Individuals were eligible to participate if they gave their written informed assent or consent, including written informed consent of all legal guardians for individuals younger than 18 years. This study follows the Strengthening the Reporting of Observational Studies in Epidemiology  is the same as the proportion of eligible participants among those with known eligibility, the estimated overall response proportion was 24.8% (American Association for Public Opinion Research formula RR3).27 The main reasons for nonparticipation were lack of time or interest, failure to contact, and arranging suitable appointments. Participation rates were higher among female participants and those with higher education.26Briefly, the sample was drawn randomly, stratified by sex and age, from the population registry of the city of Dresden, Germany, in 2015. Individuals were invited via letter (with a maximum of 2 reminder letters). Individuals were excluded if they had insufficient German language skills, were permanently institutionalized, or lived outside Dresden during the field phase. Of the 6321 invited individuals, 14.1% were known to be ineligible. Of the remaining individuals, 1180 participated in the study, resulting in a response (participation) proportion of 21.7% (American Association for Public Opinion Research formula RR1).32 with the following questions: “Have you ever thought over a period of days or weeks about killing yourself, ie, to attempt suicide?” (ideation); “Have you ever made a specific plan for killing yourself?” ; and “Have you ever attempted suicide?” .Lifetime suicidal behavior was assessed in all participants using an updated version  of the fully standardized computer-assisted Munich-Composite International Diagnostic InterviewParticipants who reported lifetime suicidal behavior were also assessed by questionnaire about age at onset, age at last occurrence, and frequency . Suicidal behavior in the past 12 months was considered to be present if the age at last occurrence for ideation, plan, or attempt was reported to have occurred in the past 12 months. Duration of suicidal behavior was defined by the time (in years) between age at onset and age at last occurrence. Time between onset of ideation, plan, and attempt was calculated as the difference between the ages at onset (in years).33 was used. Data were weighted to improve representativeness for sex and age,26 but frequencies are reported unweighted. Robust 95% CIs were calculated by using the first-order Taylor-series linearization method.34 Tests were done at the 2-sided α = .05 level. Age cohort differences (14-17 vs 18-21 years) and sex differences were examined using logistic regression analyses (odds ratios [ORs] including an F test); for count data (frequency and duration), negative binominal regression (incidence rate ratios including an F test) was used. Odds ratios were used to examine associations between duration and frequency of ideation or plan and transition.Data analysis was conducted from October 2018 to March 2019. Stata statistical software version 14.2 (StataCorp)35 This was addressed by survival analyses with event indicators  and the corresponding exit time . The Kaplan-Meier method was used to estimate age-specific cumulative lifetime incidence rates for any suicidal behavior, ideation, plan, and attempt until the age of 21 years, separately for female and male participants.36Because any participant could progress into suicidal behavior after observation and until the highest age observed (21 years), our data are right-censored.36 For ideation-to-plan, ideation-to-attempt, and plan-to-attempt transition, the time  was defined as the age difference in years between the reported ages at onset. Hence, equal age at onset reports  were treated as 0 years between ideation and plan. Age at onset of ideation and plan were used explanatorily as covariates in Cox regression analyses to examine associations with transition stratified for sex and age cohort. The proportional-hazards assumption was tested using Schoenfeld residuals.37 Once the proportional-hazards assumption was violated, separate Cox regressions were calculated for 2 age intervals before and after the (observed) interception of the curves. Only years with at least 20 individuals at risk are reported.Sex differences in cumulative incidence curves were analyzed using Cox regression analyses and quantified with hazard ratios (HRs), stratified for age.Missing values for any of the items assessing suicidal behavior (4 participants) were conservatively coded as no suicidal behavior. Two participants did not report an age at last occurrence for ideation or plan; thus, we conservatively imputed the age at onset. Similarly, 3 participants did not report an age at last occurrence for attempt; thus, the age of the most recent plan was used here as an approximation.Overall, 1180 adolescents and young adults  aged 14 to 21 years (mean [SD] age 17.9 [2.3] years) participated in the study; 97.1% reported being German citizens. Most (65.1%) lived with their parents, 12.5% lived alone, 5.4% lived with a partner, and 17.1% lived with other people. Most of the participants (76.4%) reported currently receiving or having received a high school or A-level education, 18.6% reported having a middle-school education (secondary school), and 5.0% reported attending other institutions  or having a low education level .P = .04) and plan  but not attempt . There were no significant differences between those aged 14 to 17 years vs those aged 18 to 21 years in terms of ideation , plan , or attempt . In the past 12 months, 54 participants  reported any suicidal behavior . A significant sex difference was found only for plan, indicating a higher rate among female participants  vs male participants  .Lifetime suicidal ideation, plan, and attempt were reported by 130 participants , 65 participants , and 41 participants , respectively . Any lifP = .07). The incidence curves for ideation revealed similar age patterns (P = .04). For plan, incidence was rare until age 13 years (<1%), then it increased slightly among male participants and more sharply among female participants until age 18 years  (P = .08). However, as the proportional-hazards assumption was violated , separate Cox regressions by age indicated that the cumulative incidence of attempt was similar among female and male participants before the interception point , but differed thereafter , indicating a higher incidence for female than male participants.The estimated cumulative incidence at age 21 years was 13.5% for any suicidal behavior: 12.7% for ideation, 6.6% for plan, and 4.0% for attempt  E and F.  < .001) G and H. P = .01)  and high frequency  of suicidal behavior . ApproxiP = .002) and for reporting both plan and attempt  compared with male participants. The number of plan episodes was associated with the occurrence of attempt . No association was found between duration of ideation or plan as well as frequency of ideation and the ideation-to-action transition (eTable 2 in the Two percent of the sample (26 participants) reported all types of suicidal behavior. Only ideation was reported by 59 participants (5.1%), only plan (if ideation) was reported by 39 participants (3.1%), and only attempt was reported by 8 participants (0.8%). Approximately one-half of those with ideation reported a plan , and nearly one-quarter  reported an attempt; of those who reported an attempt, 23.5% reported an unplanned and 76.5% a planned attempt . OverallThe observed time for transition ranged from 0 years  to 7 years for ideation to plan , from 0 to 4 years for ideation to attempt , and 0 to 1 year for plan to attempt .P = .003). No significant sex differences emerged for ideation-to-attempt  and plan-to-attempt  transitions.The estimated transition probability was highest in the same year . For theP = .04); that is, those with a later onset of ideation had overall a higher risk for a plan. No associations were found between age at onset of ideation  and age at onset of plan  in the transition to attempt. No association between age at onset and transition patterns were found.Age at onset of ideation was associated with plan  are as follows. First, there was a high lifetime prevalence (11.5%) and estimated cumulative incidence until age 21 years (13.5%) for any suicidal behavior. Second, suicidal behavior typically increased after the age of 10 years and most sharply at age 13 to 14 years, with different onset patterns by sex and for ideation, plan, and attempt. Third, suicidal behavior was frequently persistent and recurrent. Fourth, transition to more serious suicidal behavior was considerable: a plan was reported by 47.0% and an attempt by 23.9% of those with ideation. Approximately 17.0% to 35.1% of participants experienced an ideation-to-action transition within the same year. With these findings, the current study extends the knowledge on suicidal behavior derived by the limited number of previous studies in the critical age span of adolescence.41 including Germany.40 However, our estimates are comparable to those of studies using similar assessment methods  among participants aged 13 to 18 years from the United States4 and adults worldwide.42 The lower estimates for suicidal behavior in the present study compared with other studies could be explained by sampling procedures. Previous studies were mostly based on school-based samples that include more individuals with lower education and lower socioeconomic status,40 which seem to be associated with suicidal behavior, although results differ by type of suicidal behavior, sex, and age.43 In the study by Brunner and colleagues,39 a representative sample of 5759 ninth-grade students was studied between 2004 and 2005 in the Rhein-Neckar district in Germany, resulting in higher prevalence estimates . In contrast to previous studies, the current study revealed a female preponderance in prevalence for ideation and plan only, rather than higher rates for female than male participants in general.43 To our knowledge, no consistent findings exist for differences in prevalence by age in adolescents.43 In the present study, no age cohort differences were found, indicating that suicidal behavior is of relevance throughout adolescence.The prevalence rates in the current study are in the lower range of estimates from previous studies of adolescents around the world,44 likely as a result of both retrospective bias and real episodes in adulthood. The incidence curves in the present study are nearly identical to, although somewhat lower than, the ones presented by Nock and colleagues4 for US participants aged 13 to 18 years. Adding to the limited information about sex differences so far,4 female participants showed a higher incidence of ideation and plan during adolescence compared with male participants and for attempt from age 14 years onward in the present study. The sample size for male participants reporting an attempt was small and no new cases of plan and attempt were observed in male participants after ages 15 and 16 years. Therefore, sex differences have to be interpreted with caution. Available statistics45 clearly demonstrate a male preponderance for completed suicides, with sex difference emerging in adolescents.The occurrence of suicidal behavior throughout adolescence and young adulthood is characterized by the cumulative incidence and age at onset distribution patterns for suicidal behavior, demonstrating the high-risk period for first onset during adolescence and underlining the need for sensitive assessment throughout development. More specifically, the time from age 12 to 16 years appears to be the first crucial risk phase, with a particularly high incidence at ages 13 to 14 years. Adult samples usually report a considerable higher median age at onset, mostly in the mid-20s, for ideation, plan, and attempt,47 are available using self-reports with different time intervals for adult samples. Some studies48 used data-driven approaches, such as latent-class analyses based on dimensional measures, and found different patterns in girls and boys aged 12 to 15 years.20 The present study demonstrates both high duration  and high frequency (a median of 3 episodes over days or weeks) of suicidal behavior. Sex differences were evident only in terms of a higher number of plan episodes among male participants.Regarding temporal characteristics of suicidal behavior, so far only a few studies4 from the United States, our point estimates were higher for transition probabilities for ideation-to-plan  and lower for ideation-to-attempt . Results regarding the speed of transition are similar to previous studies indicating that the first year after onset is most critical.4 Female participants showed a higher risk for ideation-to-plan transition compared with male participants, which differs from previous findings based on similar assessments.4 The current study adds to the literature that a higher number of episodes of planning are associated with attempt. Also, age at onset of ideation seems to be relevant for the ideation-to-plan transition, with a later onset associated with a higher transition risk. With regard to timing of transition, more fine-grained studies are needed to understand the concrete pathways of suicidal behavior .Compared with the results by Nock and colleagues26 The participation rate was low. However, low participation is a general trend in recent epidemiological studies,51 especially among adolescents,53 and is not necessarily associated with the validity of the result.51 In the current study, participation bias could have had an effect on the prevalence estimates, although rates are similar to those of other studies using a similar design.1 Compared with the general population of Dresden, a smaller number of individuals with lower education participated in the study,26 which was previously associated with more suicidal behavior, although as pointed out already, the results differed by type of suicidal behavior, sex, and age,43 and this could have led to an underestimation of prevalence estimates. The sample also differed slightly from the general population in that there were more female participants. Sample weights were used to adjust for sex and age differences. The limited sample size in the current study is associated with comparatively high random error, specifically in risk estimates close to the age of 21 years and transition probabilities. Previous studies55 indicated that suicidal behavior is more common among nonresponders, so there might be an underestimation of the prevalence in the current study. Moreover, as with other epidemiological studies,54 the current study did not assess currently institutionalized individuals , who might have a higher risk for suicidal behavior. Hence, we assume that suicidal behavior is even more prevalent in the general population than our results suggest. In contrast, nonresponder analyses for the present study indicated that participation tended to be higher among those with more mental health problems,26 which could be associated with reported suicidal behavior. The presence of a mental disorder  during an interview about suicidal behavior could have had an effect on the recall of suicidal episodes, although so far little is known about the direction of this effect across different psychopathologic disorders.The present study is one of a few epidemiological studies that examined lifetime suicidal behavior in a population-based sample. Given the focus on adolescents and young adults, all results are restricted up to age 21 years and to a region  in the eastern part of Germany. Here, the population density and mean age are similar to those in most larger cities in Germany, with a lower proportion of migrant individuals.58 On the one hand, reviews60 found that individuals can recall especially adverse experiences with sufficient accuracy. On the other hand, the test-retest reliability of lifetime suicide attempt seems to be limited, with one-third of cases not reporting the attempt 4 years later32 or reporting it inconsistently over time.54 Previous studies 30 analyzing the reliability of the Composite International Diagnostic Interview time-related items, however, found generally high test-retest reliability for age at onset reports. In addition, face-to-face assessment of suicidal behavior might have resulted in fewer reports of suicidal behavior61 and, thus, in underestimated prevalence or cumulative incidence. For duration and frequency, observed data were used and censoring was not taken into account, which might have led to an underestimation of duration and frequency. By defining duration as the number of years between onset and last occurrence, it remains unknown whether suicidal behavior persisted throughout the years or rather showed a recurrent pattern with years of remission in between, which would have led to an overestimation of duration.Information assessed in the study including age at onset reports was based on retrospective reports, which might be affected by recall bias.62 may be promising new tools for recognition and intervention.Adolescence and young adulthood are critical developmental periods for the onset of suicidal behavior, with a small time frame for recognition and targeted interventions to disrupt the ideation-to-action pathway. The present study implicates, on the one hand, the need for immediate short-term interventions to prevent escalating suicidal behavior, ideally around ages 12 to 16 years, and, on the other hand, longer-term monitoring and possibly interventions to prevent or diminish the impact of persistent or recurrent patterns of suicidal behaviors across adolescence. Mobile technologies


Callosobruchus maculatus activity and economic damage on cowpea. A 15-d study was conducted with six treatments of 25, 50, and 100 C. maculatus adults in 500 and 1000 mL jars using acoustic probes inserted through stoppers sealing the jars. Acoustic activity as a result of locomotion, mating, and egg-laying was measured by identifying sound impulses with frequency spectra representative of known insect sounds, and counting trains (bursts) of impulses separated by intervals of <200 ms, that typically are produced only by insects. By the end of the first week of storage in all treatments, oxygen levels declined to levels below 4%, which has been demonstrated to cause mortality in previous studies. Concomitantly, insect sound burst rates dropped below an acoustic detection threshold of 0.02 bursts s−1, indicating that the insects had ceased feeding. Statistically significant relationships were obtained between two different measures of the acoustic activity and the residual oxygen level. Based on the experimental results, a simple equation can be used to estimate the time needed for oxygen to decline to levels that limit insect feeding damage and thus grain losses in hermetic storage containers of different insect population levels and various volumes.Acoustic monitoring was applied to consider hermetic exposure durations and oxygen levels required to stop adult  Vigna unguiculata (L.) Walp.) is an economically important indigenous protein source in the semi-arid tropical and subtropical world, namely Asia, Africa, southern Europe, and Central and South America   and (2) are statistically significant, with  < 0.05] . The regC. maculatus infesting hermetically sealed cowpea decreased over time as the oxygen level decreased. As noted in Jalinas [2 decreased below 4% during hermetic storage. From an economic perspective, insect quiescence in this study is similar to insect mortality in other studies of oxygen deprivation because the insects no longer cause damage to the stored product, even if they briefly continue to remain alive. Quiescence in response to oxygen deprivation was achieved within one week of sealing infested cowpea grain in the hermetic containers, and in a similar study examining mortality in response to oxygen deprivation in hermetic containers, the oxygen level declined from 19.2% to 2.3% within five to seven days, resulting in the death of 50 and 200 C. maculatus adults in 210 and 850 mL jars, respectively [In this study, we assessed how the activity of three different population levels of  Jalinas , reduced Jalinas  are expeectively . The speectively .C. maculatus as O2 levels decreased. Similar studies on hermetic storage have shown that the depletion rate of oxygen depends on the duration of storage, size of storage container, the developmental stage of the insects, and the level of infestation present [C. maculatus in this study also depended on the initial population level and the available oxygen. This was clearly demonstrated by the individual oxygen consumption rates for the same population level in 1000 mL treatments, which contained double the initial oxygen volume of 500 mL treatments. This scenario was also observed in a previous study with S. oryzae [The results of this study show that the initial infestation level and initial quantity of oxygen available dictated the susceptibility and survival duration of  present ,41. Indi. oryzae , whereby2 is needed during the lifetime of C. maculatus adults (egg to adult emergence) [t4%. While the insects lived for about five days under these low oxygen conditions [Other studies have assessed the individual insect consumption rates at normoxic conditions and found that approximately 8 mL Oergence) ,42,43. Onditions , the avaC. maculatus produced sound impulses with a broad range of amplitudes, spectral features, and temporal patterns, and which were comparable with those observed for other stored product pests such as Prostephanus truncatus (Coleoptera: Bostrichidae), Sitophilus zeamais (Coleoptera: Curculionidae), Acanthoscelides obtectus (Coleoptera: Chrysomelidae: Bruchinae), and Sitophilus oryzae (Coleoptera: Curculionidae) [The low oxygen levels under hermetic conditions result in the suppression of feeding, growth, development, and population expansion ,14,43,44ionidae) ,30,31,37C. maculatus observed in this study are similar to those observed in S. oryzae during hermetic storage [−1, which has previously been determined to be the threshold below which a low likelihood of infestation is predicted acoustically [.The acoustic patterns of  storage . An impostically  and, in stically . This ret4%, can be estimated for different values of initial available oxygen (AO) volume and different numbers of insects per treatment, tN, by considering the ranges of individual oxygen consumption (IOC) found in The increasing use of hermetic technologies such as PICS bags as an inexpensive, effective way to control storage insect pests in Africa and Asia ,47,48 wat4%, consider the treatment where oxygen was reduced at the lowest rate, with 25 insects in a 1000 mL hermetic container, in which t4% was measured as 11 d , yields t4% estimates of 22, 5.6, and 9.0 d, respectively. Similarly, consider the treatment where oxygen was reduced at the highest rate, with 100 insects in a 500 mL hermetic container in which t4% was measured as 2.6 d , yields t4% estimates of 2.99, 0.75, and 1.21 d, respectively. In both cases, the actual measurement occurred within the overall range of IOC measurements. Now consider that the initial AO volume of a 100 kg PICS bag is 1149 mL O2, and a typical count of insects in one of these bags could be 800 (8 adults per kg). Typical values of t4% expected for a PICS bag would then be 1149/(800 · IOC), or 8.45, 3.50, and 2.18 d, respectively.As an example of how effectively the approximation reproduces the measured value of  as 11 d B. Calculas 2.6 d A. ApplyiIOC), available oxygen (AO), and starting population level (tN) before hermetic storage could be beneficial in understanding how fast oxygen is depleted to levels where insects can no longer damage the grain. Acoustic activity cessation and overall quiescence of insects during hermetic storage occurs at oxygen levels below 4% and is a good indicator of the safety level of stored grain. Knowledge of the dynamics within the hermetic enclosures heightens the appeal to use hermetic storage as opposed to conventional non-hermetic storage methods.Overall, the relationship between individual insect oxygen consumption of the insects (


We investigate the interocular symmetry of fixation, optic disc, and corneal astigmatism in bilateral high myopia, and evaluate the predictive relationships between them.We enrolled 202 cases with bilateral high myopia. Fixation, in terms of the bivariate contour ellipse area (BCEA), was evaluated with the Macular Integrity Assessment microperimetry. Optic disc features, including orientation, tilt, and rotation, were evaluated with ultrawide-field retinal photographs. Corneal topography was performed with Pentacam. Interocular symmetry of fixation, optic disc, and corneal astigmatism was assessed, and the predictive relationships between these parameters were investigated.P < 0.05). No interocular symmetry was observed in the direction of the fixation ellipse. In both eyes, the corneal steep meridian more often was consistent with the optic disc orientation than inconsistent .Axial length differences between the two eyes were: ≥0 to ≤1 mm, 67.8%; 1 to ≤2 mm, 20.3%; 2 to ≤3 mm, 9.4%; and >3 mm, 2.5%. Axial length, 95% BCEA, and magnitude of corneal astigmatism showed good interocular symmetry, whereas the optic disc tilt, rotation, and axis of corneal astigmatism (mirror axes) showed less symmetry (all As different parameters presented different degrees of symmetry, cautions are needed when including both eyes or only one lateral eye in cases of bilateral high myopia for clinical investigations. The optic disc orientation, to some extent, may indicate the steep meridian of the cornea.Our study provided evidences for selection of eye laterality in clinical investigations of highly myopic eyes. High myopia is a major worldwide concern today, especially in East Asia.5Several studies have investigated variations in optic disc features, such as tilt, rotation, and parapapillary atrophy, in highly myopic eyes.611Previously, we identified an axial length–dependent reduction in fixation stability of highly myopic eyes using the Macular Integrity Assessment (MAIA) microperimetry system .Such great variations in these anatomical or functional parameters of highly myopic eyes may inevitably increase the interocular asymmetry in patients with bilateral high myopia. We normally expect eyes and visual systems to behave somewhat symmetrically, and accordingly, we include both eyes of patients for many clinical investigations. However, this may not be correct, especially in highly myopic eyes, which show great structural viability due to abnormal elongation of the eyeballs. Studies have reported a consistent pattern of asymmetry between our eyes. The Sydney Childhood Eye Study found only a low level of interocular correlation in the thickness of the retinal nerve fiber layer (RNFL), with differences of up to 17 μm between the two eyes of normal children.Additionally, we assume that if the eyeball is an ideal optical system, certain kinds of predictive relationships may exist between the cornea astigmatism median and optic disc orientation or the direction of the macular fixation ellipse, to maintain the coordination of eye function. A previous study showed that an abnormally shaped optic disc can be correlated with abnormal corneal astigmatism and that the steep meridian of corneal astigmatism may reflect the orientation of the longest disc diameter.www.clinicaltrials.gov (accession number NCT03062085).The Shanghai High Myopia Study is a hospital-based prospective cohort study that continuously includes highly myopic patients scheduled for cataract surgery in the Eye and Ear, Nose, and Throat (EENT) Hospital of Fudan University since October 2015. All subjects underwent detailed preoperative examinations and postoperative follow-ups. This investigation is a subproject of the Shanghai High Myopia Study. Individuals with bilateral axial lengths >26 mm were selected for this study; excluded were those whose eyes had severe myopic fundus pathologies, such as choroidal neovascularization, macular hole, or severe macular atrophy, a history of other anterior segment ocular diseases, trauma, or a systemic condition that affected their visual performance. Ultimately, 202 subjects were eligible for this study. The institutional review board of the EENT Hospital of Fudan University approved the study protocol. All procedures were performed in accordance with the tenets of the Declaration of Helsinki. All participants were informed of the study objectives, and each signed an informed consent form to participate in this study. This study was registered at Preoperative examinations included assessment of visual acuity, tonometry, measurement of axial length , funduscopy, B scans, and macular scans with optical coherence tomography . Corneal topography was performed with the Pentacam system , and the steep meridian of the anterior corneal astigmatism was recorded. Follow-up included assessment of visual acuity, manifest refraction, tonometry, funduscopy, retinal photography, MAIA microperimetry, and an OCT macular scan.2) of the ellipse containing most of the fixation positions registered during the measurement procedure, and the fixation ellipse angle, the orientation of the longest ellipse diameter. BCEA normally is calculated by considering 63% or 95% of the fixation points. Because these two parameters are similar, only 95% BCEA was used for further analysis in this study.Fixation was assessed using the MAIA microperimetry system. The patients were asked to stare at the fixation stimulus, which consisted of a red circle with a diameter of 1°. The eye trackers within the MAIA system detected fixation loss as a misalignment of the directions of the central fixation stimulus and gaze, and recorded the points of fixation. The device then automatically calculated two parameters of fixation: the BCEA, which refers to the area  with standard settings. The measurements of optic disc tilt and rotation were obtained as described previously.t-test was used to compare continuous variables and the χ2 test was used to compare categorical variables of two eyes. Intraclass correlation coefficients (ICC) and Pearson's coefficients were computed to measure the interocular agreement or correlation. The relationships between the cumulative distribution functions were assessed with the Kolmogorov–Smirnov test. P < 0.05 was considered statistically significant.The agreement between two observers on the optic disc orientation, tilt ratio, and degree of rotation was evaluated with the Bland–Altman method, which plots their mean results against their differences . The limt-test, all P > 0.05), except that the left eyes had significantly higher optic tilt ratios than the right eyes . The characteristics of the subjects are shown in P < 0.05). The optic disc tilt, rotation and steep corneal meridian (axis of the right eye and mirror axis of the left eye) showed low to moderate correlations , indicating interocular asymmetry increased. The findings were similar with Pearson's correlation analysis .We firstly investigated interocular symmetry. The distributions of the differences in axial length between the two eyes were: ≥0 to ≤1 mm, 67.8% (137/202); 1 to ≤2 mm, 20.3% (41/202); 2 to ≤3 mm, 9.4% (19/202); and >3 mm, 2.5% (5/202). analysis . HoweverP = 0.003; left eye, P = 0.021; cornea vs disc, both eyes, P < 0.001).From the cumulative distribution function curves shown in χ2 test, right eye, P < 0.001; left eye, P = 0.029). However, no such consistency was detected between the steep corneal meridian and fixation ellipse angle in either eyes . In addition, the proportion of the angles between the steep corneal meridian and optic disc orientation that were <10° was 14.4% (29/202) in the right eye and 11.9% (24/202) in the left eye, and the proportion of the angles between the steep corneal meridian and fixation ellipse angle that were <10° was 13.9% (28/202) and 9.4% (19/202), respectively. If the angle range was set at <45°, the percentage was 55.9% (113/202) and 50.5% (102/202), respectively, for the former, and 49.5% (100/202) and 46.5% (94/202), respectively, for the latter.We then evaluated the predictive relationships between the direction of the fixation ellipse, orientation of the longest disc diameter, and steep corneal meridian. We defined 0° to 45° and 135° to 180° as the horizontal zone and 45 to 135° as the vertical zone. If the steep corneal meridian and optic disc orientation or the fixation ellipse angle were located in the same zone, their directions were considered consistent. The steep corneal meridian more often was consistent with the optic disc orientation than inconsistent in the right and left eyes,  and mirror (mirror axes) symmetry. McKendrick et al.Fixation is a recently proposed concept in the study of ocular function. In healthy eyes, small fixational eye movements in the right and left eyes are not absolutely symmetrical, and this can be quantified using the BCEA measured with MAIA microperimetry.The structural and functional symmetry in highly myopic eyes has important clinical significance. High myopia is an oculopathy that causes a series of anatomic and pathologic changes, which may consequently induce greater asymmetry between the right and left eyes. This possibility challenges the reliability of studies that enroll only one lateral of eyes or those that mixed the data of two eyes. Bias also will be introduced that impairs the reference values for ocular parameters taken from normative databases. Bilateral asymmetry in ocular metrics also might affect the occurrence of certain ocular diseases. There is asymmetry in the timing of progression of age-related macular degeneration, as reported by the researchers in the Beaver Dam Eye Study.The predictive relationships between the anterior and posterior poles of the eyeball also are important, because they affect the coordination of the optical system, as in architectonics, in which a correspondence between the roof and ground floor structures is required. A previous study proposed that the steep corneal meridian is associated with the orientation of the longest disc diameter;To conclude, in patients with bilateral high myopia, different parameters presented different degrees of symmetry. Therefore, cautions are needed in interpreting data from clinical investigations, which include both eyes or only one lateral eye in cases of bilateral high myopia. The optic disc orientation may, to some extent, indicate the steep meridian of the cornea.Supplement 1Click here for additional data file.


Xerocomus chrysenteron , Clitocybe nebularis, Lepista nuda and Lactarius semisanguifluus . Poisonous mushrooms included Amanita phalloides , Boletus satanas (n = 3), Amanita muscaria (n = 2) and Amanita pantherina (n = 2). There were no fatalities and 80% of the patients were discharged within 24 h. Mushroom poisoning does not appear to be a common reason for emergency consultation and most presentations were of minor severity and related to edible species . Nevertheless, poisonous mushrooms and severe complications were also recorded. Collaboration with a poison centre and/or mycologists is of great importance, especially in high risk cases.The consequences of mushroom poisoning range from mild, mostly gastrointestinal, disturbances to organ failure or even death. This retrospective study describes presentations related to mushroom poisoning at an emergency department in Bern (Switzerland) from January 2001 to October 2017. Gastrointestinal disturbances were reported in 86% of the 51 cases. The National Poisons Information Centre and mycologists were involved in 69% and 61% of the cases, respectively. Identification of the mushroom type/family was possible in 43% of the cases. The most common mushroom family was Boletaceae (n = 21) and the most common mushrooms  Of the estimated 5000 existing mushroom species, only 200–300 have been established to be safely edible, while 50–100 are known to be poisonous to humans and the toxicity profile of most other species has not been investigated ,5. PoisoTox Info Suisse provides 24-h medical advice nationwide in cases of (suspected) poisoning. In recent decades, there has been a steady growth in mushroom-related calls, with an autumnal peak seen each year [Tox Info Suisse, the Swiss Mushroom Control Association  offers checkpoints for a voluntary assessment of self-harvested mushrooms, in order to check edibility. In 2014, 4504 harvests were checked by the French-speaking part of VAPKO; 37.9% of those contained inedible, 12.5% poisonous and 1.7% lethal mushroom species [In Switzerland, the National Poisons Information Centre ach year . In 2016ach year . The mosach year . In addi species .Despite possible regional differences, only a study from the French speaking part of Switzerland has investigated the epidemiology and clinical characteristics of patients presenting with mushroom poisoning at an emergency department (ED) in Switzerland , and theThis was a retrospective single centre study of patients presenting at the ED of Bern University Hospital  with signs/symptoms related to mushroom exposure or intoxication during a 17 year period (from 1 January 2001 to 31 October 2017). The ED of the Inselspital is both a primary care facility  and a tertiary referral centre for other hospitals in the region and provides a catchment area for approximately two million people—with about 46,000 (2017) emergencies admissions a year (≥16 years of age).Patients admitted to the ED are registered in a specific clinical program: The program Qualicare  was used for the period from 1 January 2001 to 31 May 2012 and a second program—E.care —from 1 June 2012 to 31 October 2017. These databases were used to identify all patients presenting with (suspected) mushroom poisoning. Cases were retrieved using a comprehensive full-text search algorithm with mushroom poisoning, related terms and a large number of specific mushroom names, including their German translation, as search terms. Each identified case was reviewed by one of the authors of the study. We included each patient requiring medical evaluation at the ED for mushroom poisoning, regardless of the circumstances of exposure  and the severity of the complaints. We excluded patients with no history of mushroom consumption and cases where the complaints could be clearly attributed to a cause other than mushroom poisoning based on the available final diagnosis in the medical report and the documented evaluation of the physician assessing the patient.Tox Info Suisse, treatment provided, type of discharge from the ED , length of hospital stay, and outcome . As in previous studies [The following parameters were extracted (if available) from the charts of included patients: age, sex, type of transport to the ED , date of admission and discharge, mushroom species and how they were obtained , circumstances of exposure , symptoms, interval from ingestion to onset of symptoms, laboratory findings (including detection of α-amanitin in urine), consultation of a mycologist or  studies ,9, we di studies , i.e., p studies . Data weDuring the study period (1 January 2001 to 31 October 2017), 62 patients were admitted to the ED of Inselspital due to (suspected) mushroom poisoning. Eleven patients were excluded because mushroom consumption could not be confirmed, or the complaints could clearly be attributed to a cause other than mushroom poisoning. Finally, 51 patients were included in our study. The mean (±SD) age of the study population was 48.4 ± 16.9 years (median 48 years), with a range from 15–79 years. Twenty-nine patients (56.9%) were female and 22 (43.1%) male. The annual number of presentations over the study period ranged from zero to ten patients, with a peak in 2008 . The seaTox Info Suisse, air rescue service or internal allocation. Mycologists were involved in 60.8% (n = 31) of the cases and Tox Info Suisse was contacted for 35 patients (68.6%) ( (68.6%) . The exaIn 74.5% of the cases (n = 38), the mushrooms were self-harvested and in 94.1% of the cases (n = 48) exposure was accidental. Intentional ingestion included recreational use in two cases  and a suicide attempt in one case . In most cases, the symptoms of mushroom poisoning were gastrointestinal disturbances; further reported symptoms included neurological, psychiatric, and cardiovascular disorders .Among the 48 symptomatic patients (94.1%), 35 patients (68.6%) presented with early symptoms  and 13 patients (25.5%) with delayed symptoms  .Amanita phallloides); 31.4% of the cases (n = 16) had normal laboratory parameters and in 15.7% (n = 8) no laboratory data were available . One of these patients (ID 14), developed acute renal failure on the 4th day and elevated liver enzymes with a peak on the 6th day , despite normal laboratory parameters during the first three days (Boletus satanas (In 52.9% of the cases (n = 27), laboratory abnormalities were found with various deviations from the normal range . Elevatevailable . In 37.3vailable . In the ree days . The acu satanas . After f satanas .N-acetylcysteine, and four  with penicillin G. Seven patients received no treatment, either because they were asymptomatic, or due to spontaneous resolution of their symptoms. Gastric and jejunal tubes were used in one case due to prolonged diarrhoea and vomiting (In 84.3% of the cases (43 patients), treatment included intravenous fluids . Symptomatic therapy included metoclopramide , butylscopolamine , meclozine/pyridoxine, domperidone, ondansetron, esomeprazole and paracetamol . Twenty-nine patients received activated charcoal (56.9%); 16 patients  were treated with silibinin, 12 patients  with vomiting . All patAlthough the total number of cluster patients was 37, data were not available for ten of those patients who did not consult our ED. Therefore, 27 of the cluster patients are described in our study . In three quarters of the cases, the mushrooms were self-harvested. A mycologist and/or Tox Info Suisse [Amanita phalloides, Boletus satanas, Amanita muscaria and Amanita pantherina. Among the main mushroom toxins categories, gastrointestinal irritants were the most common in our study, followed by neurotoxins and protoplasmic poisons. Boletaceae was the most commonly identified mushroom family in our study, as in the report from o Suisse  and the o Suisse , which mo Suisse ,29, mostA check list of some important points regarding patient history and management in cases of (suspected) mushroom poisoning is contained in the supplement  or only moderate (ID 19) transaminase elevation. Despite this, those patients were hospitalised for several days, most probably to assure regular follow-ups in order to exclude a delay onset hepatic and/or multisystem organ failure. Although not routinely available, analytical methods such as urinary ELISA can be used to detect amatoxin in cases of suspected amatoxin poisoning. This method has good sensitivity up to 100%), especially if performed within 36 h after ingestion ,36, alth00%, espeoisoning , ID 14, Amanita phalloides poisoning includes administration of activated charcoal for gastrointestinal decontamination and silibinin [N-acetylcysteine, similarly to paracetamol intoxication [Amanita phalloides ingestion received activated charcoal, silibinin and N-acetylcysteine. Despite the current recommendation, penicillin G was administered in combination with silibinin in four cases in our study, which might be because older guidelines were applied . Furthermore, since some patients with suspected poisoning received silibinin as precautionary measure while awaiting the analytical results, an in-house test could contribute to avoiding unnecessary administration of drugs and reducing length of hospital stay.Treatment of ilibinin , a milk ilibinin ,38,39 anilibinin . The comilibinin . In casexication ,40. In oSimilarly to other studies ,4,29, inTox Info Suisse/mycologists. The authors of the study in Italy [Identification of the specific mushroom(s) involved can contribute to appropriate diagnostic and treatment. In cases of mushroom poisoning, the exact mushroom species is successfully identified in the range of 5–27% of patients ,31, within Italy  also repTox Info Suisse also shows marked seasonal fluctuation [Similarly to our findings, analysis of mushroom poisoning data from ctuation  and mostctuation ,13,16,29Tox Info Suisse/mycologist before the analytical results were available, since the exact time a mushroom specialist was contacted is not known. As a result, it is also not possible to describe how the patient management was adjusted  based on the mushroom specialist consultation in each case. Finally, our data may not be generalised to other regions in Switzerland, inter alia due to differences in local weather. The main strengths of our study include its long duration of 17 years. In addition, we frequently consulted a mycologist and Tox Info Suisse and this contributed to our high rate of identification of the mushrooms.The limitations of our study include the retrospective design. Although we aimed to identify all mushroom poisoning cases during the study period using the extensive search of our emergency unit specific software, it is possible that some cases were missed, e.g., if a patient was falsely diagnosed or did not mention the mushroom consumption, or that in some cases the symptoms were attributable to another cause that remained unrecognized despite thorough diagnostic at the ED. Moreover, although it is to assume that the patients remained free of symptoms if there was no re-presentation following their discharge, no information was available regarding their further outcome after leaving the ED. It is also possible that in some cases the treatment did not contribute to the clinical improvement . Thus, it is not possible to provide an accurate evaluation of the undertaken therapeutic measures, Furthermore, it is possible that the diagnosis of mushroom poisoning was made after discharge from the ED or that patients from peripheral hospitals were directly admitted to a ward, thus resulting in cases that could not be found in our ED database, and it is also possible that some patients developed symptoms or complications after being discharged. Moreover, we cannot report in how many cases identification or exclusion of amatoxin-containing mushrooms was provided by the Based on our findings, ED consultations due to mushroom poisoning do not seem to be very common and mostly include mild symptoms associated with gastrointestinal irritants, usually with favorable outcome. However, although less common, cases of poisonings with potential lethal mushrooms and severe complications were also recorded. Collaboration with the local poison centre and a mycologist is of great importance for the identification of the mushroom(s) involved and decisions on patient management, especially in cases of high risk. The data of our study can contribute to the development of preventive strategies, such as medical personnel training and information material for the public  on the correct handling of and potentially life-threatening mushrooms, especially during the high risk season for mushroom poisoning.


Type 2 diabetes mellitus (T2DM) is a common, chronic disease that is closely associated with anthropometric indices related to obesity. However, no study published to date has simultaneously examined the associations of T2DM with anthropometrics, bone mineral density (BMD), and body composition variables. The present study aimed to evaluate the associations of T2DM with anthropometrics, BMD and body composition variables and to identify the best indicator of T2DM in Korean adults.The data used in this study were obtained from the Korea National Health and Nutrition Examination Survey conducted from 2008 to 2011. A total of 7,835 participants aged from 40 to 90 years were included in this study. A binary logistic regression analysis was performed to examine the significance of differences between the groups with and without T2DM, and the areas under the receiver operating characteristic (AUCs) curves were calculated to compare the predictive power of all variables.In men, waist-to-height ratio (WHtR) displayed the strongest association with T2DM (adjusted odds ratio (OR) = 1.838 [1.513–2.233], adjusted p<0.001), and waist circumference (WC) and WHtR were the best indicators  of T2DM among all the variables. In women, left leg (LL) and right leg (RL) fat displayed strong negative associations with T2DM , and WC and WHtR were excellent indicators  of T2DM among all the variables. In particular, the WHtR in men and LL and RL fat in women exhibited the strongest associations with T2DM, and the predictive power of the WC and WHtR was stronger than BMD, fat, and muscle mass variables in both men and women. Additionally, the predictive power of the WC and WHtR was stronger in women than in men.Of the anthropometric indices, BMD, and body fat and muscle variables, the best indicators of T2DM were WC and WHtR in both Korean men and women. The results of the present investigation will provide basic information for clinical studies of patients with T2DM and evidence for the prevention and management of T2DM. Diabetes is a common, chronic disease worldwide, and the number of patients continues to increase due to reduced physical activity and increased obesity . The WorMany studies to date have assessed the association of T2DM with anthropometric indices, bone mineral density (BMD), and body composition variables. Among the anthropometric indices, many studies have reported the strongest association between the waist-to-height ratio (WHtR) and T2DM –15. AccoThe data used in this study were obtained from the 2008–2011 Korea National Health and Nutrition Examination Survey , which is a prospective, cross-sectional, nationally representative survey study conducted by the Korea Centers for Disease Control and Prevention ,44. The Health interviews and health examinations included in the KNHANES were performed at mobile examination centers (MECs). The health interviews, which requested information such as medical conditions, housing characteristics, socioeconomic status and alcohol use, were collected by a face-to-face interview and self-administered form. Health examinations requesting information such as height, weight, blood test and BMD were also measured at MECs .The KNHANES I—VII were conducted from 1998 to 2016. BMD examinations were performed only from 2008 to 2011; therefore, we chose the KNHANES IV-2, 3 and V-1, 2 to find associations of T2DM with various bone and body composition variables. The KNHANES IV-2, 3 and V-1, 2 included 21,303 participants who completed blood tests, bone densitometry and body fat composition tests, along with a health examination. According to the Korean Diabetes Association ,46. TherDiabetes mellitus is defined as a metabolic disease characterized by hyperglycemia resulting from defects in insulin secretion, insulin action, or both . WHO and2). WHtR was calculated as WC divided by height. Blood samples were collected from the antecubital vein after an 8-hour fast. Fasting plasma glucose, total cholesterol, triglyceride , and high-density lipoprotein-cholesterol (HDL) levels in all participants were measured enzymatically using a Hitachi Automatic Analyzer 7600 . The visceral adiposity index (VAI), a gender-specific index based on WC, BMI, TG and HDL, was calculated as follows ) and women (AUC = 0.747 [0.728–0.766]).T2DM is a representative metabolic disease associated with various diseases, such as hypertension ,50, hearOur findings indicate that WC and WHtR are the best indicators of T2DM in Korean adults and showed that the power of general anthropometric indices for the identification of T2DM was better than those of BMD and body composition variable. The WC and WHtR are known as obesity indicators. Numerous studies have investigated the association of obesity indicators with T2DM and attempted to identify the best indicators of T2DM. As shown in the study by Tulloch-Reid and colleagues , BMI andIn addition, diabetes affects bone metabolism –32. We aSome studies have investigated gender differences in correlations between T2DM and obesity measures , such asThe present study had several limitations. First, cause-effect associations were difficult to determine because of the cross-sectional design. Second, our results were limited to Korean adults because we used data from the KNHANES in this study. Third, since KNHANES does not contain detailed information on medication use, a more precise analysis was not possible. Fourth, a large number of values were missing in the original data samples, resulting in the exclusion of a number of the samples during the sample selection process, and only 36.78% of samples were selected. Therefore, we were unable to conclude that the results of this paper represent the frequency of T2DM in Koreans. Fifth, the datasets from KNHANES may be affected by response bias because self-reported data from participants such as "the current use of antidiabetic medications or glucose-lowering drugs, or physician-diagnosed diabetes" are also used in the diagnosis of T2DM.In the present study, we evaluated the associations of T2DM with anthropometrics, BMD, body fat mass and muscle mass in Korean adults. The WHtR in men and LL and RL fat in women displayed the strongest associations with T2DM, and the WC and WHtR in both men and women exhibited the highest predictive power for T2DM. Moreover, age, VAI, WC and WHtR exhibited greater predictive power in women than in men. Our findings provide clinical information that may identify patients with T2DM during initial screening steps. Further studies are needed to build a model for accurate identification based on a combination of BMD, anthropometric, and fat mass data.