Streptococcaceae, Lactobacillaceae, and Prevotellaceae were positively related to finishing weight, whereas the microbial taxa associated with intestinal inflammation and damage exhibited opposite effects. In addition, interactions of these microbial species were found to be linked with finishing weight for the first time. Gut microbial functional annotation analysis indicated that CAZymes, such as glucosidase and glucanase could significantly affect finishing weight, given their roles in increasing nutrient absorption efficiency. Kyoto Encyclopedia of Genes and Genomes (KEGG) Orthologies (KOs) and KEGG pathways analysis indicated that glycolysis/gluconeogenesis, phosphotransferase system (PTS), secondary bile acid biosynthesis, ABC transporters, sulfur metabolism, and one carbon pool by folate could act as key factors in regulating finishing weight. Additionally, SCFA levels, especially acetate and butyrate, had pivotal impacts on finishing weight. Finishing weight-associated species Prevotella sp. RS2, Ruminococcus sp. AF31-14BH and Lactobacillus pontis showed positive associations with butyrate concentration, and Paraprevotella xylaniphila and Bacteroides sp. OF04-15BH were positively related to acetate level. Taken together, our study provides essential knowledge for manipulating gut microbiomes to improve finishing weight. The underlying mechanisms of how gut microbiome and SCFAs modulate pigs’ finishing weight required further elucidation.Finishing weight is a key economic trait in the domestic pig industry. Evidence has linked the gut microbiota and SCFAs to health and production performance in pigs. Nevertheless, for Diannan small ear (DSE) pigs, a specific pig breed in China, the potential effect of gut microbiota and SCFAs on their finishing weight remains unclear. Herein, based on the data of the 16S ribosomal RNA gene and metagenomic sequencing analysis, we found that 13 OTUs could be potential biomarkers and 19 microbial species were associated with finishing weight. Among these, carbohydrate-decomposing bacteria of the families  Domesticated pigs account for a high proportion of meat production in the world and also serve as an important animal model for basic biomedical research studies . The swiRuminococcaceae and Lachnospiraceae could generate a high concentration of butyrate, which is responsible for the higher feed intake and body weight of broilers  and the top six pigs with the highest body weight (high group) for both 16S rRNA gene sequencing and metagenomic sequencing. The normal distribution of finishing weight for all DSE pigs was displayed in Supplementary Figure S1Supplementary Figure S1p < 0.01). All DSE pigs were healthy and not fed with probiotics, prebiotics, antibiotics, and anti-coccidial drugs throughout the whole experimental period. The fresh feces of the DSE population were collected directly from the rectum. All samples were stored in the 2-ml sterilized plastic sampling tubes and then dipped into liquid nitrogen immediately. After long-distance transportation, samples were stored in a refrigerator (−80°C) until processing.One hundred and two Diannan small-ear (DSE) pigs  with similar weaning weights were grouped into different commercial pens and bred under the same nutritional and management conditions until the finishing stage (345 ± 3 days). At the finishing stage, all DSE pigs were fed with same diet two times 1 day using the compound feeds containing 14–16% of crude protein, 0.25–0.60% of sodium chloride, 0.60–1.50% of calcium, 8% of coarse fiber, 3,100–3,200 kJ of digestible energy, and 0.70% of lysine, and offered an The QIAamp fast DNA stool Mini Kit  was selected to extract microbial DNA based on the manufacturer’s guidelines. The concentration and quality of stool DNA were assessed by spectrophotometer Nanodrop 2000  and 1.5% agarose gel electrophoresis, respectively. The specific primer 338F (5′-ACTCCTACGGGAGGCAGCA-3′) and 806R (5′-GGACTACHVGGGTWTCTAAT-3′) was used to amply the hypervariable regions (V3–V4) for 16S rRNA gene. The PCR amplification was accomplished by following steps: initial 94°C denaturation step for 5 min, subsequently, 30 cycles of 95°C for 30 s, 55°C for 25 s, and 72°C for 25 s followed by a final extension step for 10 min at 72°C. All amplicons were sequenced using the paired-end method on a MiSeq platform  following the standard protocols. The quality control process of raw data was performed by using QIIME (v.1.9.1), including removing primers, barcodes, and low-quality sequences . FLASH . Before To identify the biomarker characteristics among low and high finishing weight groups, linear discriminant analysis effect size LEfSe;  was perfvia MOCAT  DNA library was established for each sample, and sequencing was implemented on the Illumina HiSeq 4000 platform. Quality control, adapter trimming, and low-quality read removal of sequenced data were proceeded by software fastp  according to the approaches previously described with small modifications . A totalp value of more than 0.05 were excluded, which allows us to only focus on those species that strongly co-occurred and were more likely to affect each other. The major modules in the interaction network were calculated and visualized using Gephi software . After that, Spearman’s correlation analysis was used to test for the correlations between finishing weight phenotypic value and modules.To construct the interaction network of all finishing weight associated species, the pair-wised Spearman correlations analysis was performed between these species. Spearman’s correlation coefficient of less than 0.65 and adjust t-test (unpaired) with FDR correction between low and high finishing weight pigs. Interactive Tree Of Life (iTOL) was used to visualize the differential metagenomic species, and the other charts were finished by R software.The differential metagenomic species and function capacities for the gut microbiome between low and high finishing pigs were detected by the Wilcoxon test with FDR correction. Statistical comparisons of SCFAs in stool samples were done using the two-sided Student’s p < 0.01). In addition, the same tendency was also observed in the Chao index  and richness (Chao) indices for alpha-diversity were compared to discover the difference for the gut microbial community. As shown in p < 0.01).Principal Coordinate Analysis (PCoA) was performed to observe the change of gut microbial community structures between two experimental populations. The distribution of microbiota between the two groups was clustered separately along principal coordinates . The anaFirmicutes, Bacteroidetes, and Proteobacteria were the top three dominant phyla . For low finishing weight group, Desulfovibrio piger and Desulfovibrio sp. An276 from the family Desulfovibrionaceae were the most abundant species. In addition, three species belong to the family Clostridiaceae, including Clostridium perfringens, Clostridium sp. CAG:349, and Clostridium paraputrificum were abundant in pigs with low finishing weight. Three species from the member of family Corynebacteriaceae, named Corynebacterium freneyi, Corynebacterium halotolerans, and Corynebacterium humireducens were also enriched in the low finishing weight group. Conversely, we found that Streptococcus equinus, Streptococcus salivarius, and Streptococcus lutetiensis (members of the family Streptococcaceae) were augmented in high finishing weight group, followed by Lactobacillus reuteri, Lactobacillus pontis, Lactobacillus plantarum, and Lactobacillus equigenerosi. Moreover, Prevotella sp. RS2, Ruminococcus sp. AF31-14BH, Paraprevotella xylaniphila, and Bacteroides sp. OF04-15BH were also abundant in the high finishing weight group.At the species level, 16S rRNA gene sequencing possesses large limitations in both accuracy and resolution. Herein, we performed metagenomic sequencing for 12 individuals to further identify the differential species between low and high finishing weight groups. A total of 19 species showed different enrichments among two groups identified by the Wilcoxon rank-sum test with FDR correction using metagenomic sequencing data . Among these, 11 CAZymes especially glucosidase and glucanase , lytic cellulose monooxygenase , and Glucosyltransferase  were significantly enriched in high finishing weight group. While regarding low finishing weight pigs, we found that mannosyltransferase , peptidoglycan lytic transglycosylase, and levansucrase had significantly higher abundances.Twenty-one CAZymes with significantly different abundances were detected (p < 0.05). We found that one carbon pool by folate, limonene, and pinene degradation, and tryptophan metabolism were more active in the gut microbial communities of the low finishing weight group. Meanwhile, in pigs with high finishing weight, gut microbiota was more capable of operating glycolysis/gluconeogenesis, degradation of aromatic compounds, phosphotransferase system (PTS), glutathione metabolism, and secondary bile acid biosynthesis.A total of nine differential enriched KEGG pathways were detected in the gut microbiome of pigs with distinct finishing weights (p < 0.05). Eight KOs mainly related to ABC transporters , Fatty acid degradation , and Sulfur metabolism  were plentiful in low finishing weight group. The other 15 KOs were more abundant in high finishing weight group, most of which were associated with glycolysis/gluconeogenesis , carbohydrate metabolism , lipid metabolism:, and the biosynthesis of secondary metabolites .On the other hand, 23 KOs showed distinct abundances in pigs with low and high finishing weights (p < 0.05 and p < 0.01). No significant change was found in the levels of propionic acid between the two groups  in feeds and possesses beneficial roles in intestinal health by regulating the immune-inflammatory system , which were involved mainly in glucose metabolism and lipid accumulation  for children who consumed high carbohydrate and saturated fat diets was found in a recent investigation  as a product  and butyrate-producer  through regulating host appetite and feeding behavior  bacteria (Streptococcus was associated with lipid and bile acid homeostasis in the gut (Desulfovibrio) has been demonstrated to be associated with inflammatory bowel diseases in mice  mice/minipigs’ intervention experiment and to obtain more mechanistic understandings into the cross-talk between gut microbes and host and its effects on growth performance of DSE pigs by multi-omics sequencing analysis are needed. In addition, supplementing the microbial species from Lactobacillus spp., Streptococcus spp., and Prevotella spp., or SCFAs  into the basal diet might be a valuable strategy to improve the finishing weight of DSE pigs.Our study, although limited by a small pig population, offers important information that upholds the potential of intestinal flora manipulation in improving the finishing weight of DSE pigs. Nevertheless, future studies aiming to validate the positive or negative role of certain bacterial species in regulating finishing weight In summary, the present study detected some key microbial taxa related to finishing weight. We also performed functional annotation analysis and identified several finishing weight-associated CAZymes, KOs, and KEGG pathways. Furthermore, we highlighted the important role of SCFAs in finishing weight, especially acetate and butyrate. Hence, our investigation provides essential insights into the microbe-SCFAs interplay in affecting finishing weight and hints that modulating the intestinal bacterial communities could be an effective scheme to ameliorate finishing weight for DSE pigs and possibly other pig breeds.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: NCBI BioProject—PRJNA910336.The animal study was reviewed and approved by Animal Care and Use Committee (ACUC) in Hunan Agricultural University .ML and YY conceived and designed the experiments, supervised the experiment progress, and revised the manuscript. YL, PP, LY, and HZ helped to collect the samples. QL performed the experiments, analyzed the data, and wrote the manuscript. PH, HM, and HW revised the manuscript. All authors contributed to the article and approved the submitted version.This study was supported by grants from the Natural Science Foundation of Hunan Province (No. 2022JJ40171), National Natural Science Foundation of China (No. 32002166), Breeding and Promotion of Lancang black pig (2021kjc-js072), Key Science and Technology Project of Yunnan Province (202202AE090032), Consulting research program of Yunnan Province pig industry development strategy (2022YNZH7-2), and the Science and technology innovation Program of Hunan Province (2021RC3091).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Although there are very effective treatment approaches for Attention Deficit Hyperactivity Disorder (ADHD) available, the clinical management has its limits making new treatment modalities a necessity. Evidence suggests that low frequency repetitive transcranial magnetic stimulation (rTMS), improves hyperactivity, impulsivity, and attention deficit of individuals with ADHD.The aim of this case study is to present the effectiveness of rTMS on hyperactivity, impulsivity, and attention deficit of an individual with ADHD.This is a case study of a 22 year old male diagnosed with ADHD. The protocol applied was 2 weeks of daily rTMS sessions. This involved repetitive TMS to the right dorsomedial prefrontal cortex  to treat attention deficit, hyperactivity and impulsivity. Assessments were conducted using Adult ADHD Questionnaires, the Jasper and Goldberg questionnaire and WHO self- reported scale at baseline, 1 month and up to 3 months follow up.The patient showed overall improvement in scores in both ADHD scales with scores dropping more than 50 % in both scales from pre treatment to 3 months follow up. No side effects were recorded during therapyOur findings suggest that the use of rTMS therapy in individuals with ADHD, is an efficacious and safe therapeutic treatment option.


Attention Deficit Hyperactivity Disorder (ADHD) is a common neurodevelopmental disorder often diagnosed between the ages of 7 and 10. The estimated prevalence of ADHD in adults is usually 2-5%, which means that in up to half of people this diagnosis persists into adulthood.The aim of this sub-study was to determine whether there are significant differences in ADHD symptomatology between individuals who have taken or are taking prescription drugs and those who have never taken them.Data collection was performed by STEM / MARK in January 2019 through the European National Panel. Respondents completed a demographic questionnaire focusing on the history of ADHD and a standardized ADHD Self-Report Scale (ASRS) questionnaire for the symptomatology of ADHD in adulthood.Of the 1,518 respondents, 3% reported being diagnosed with ADHD / hyperkinetic disorder during their lifetime. According to the ASRS assessment, 119 respondents were classified as suspected ADHD. Overall, men scored higher symptoms of ADHD. The results also show that the group that states that taking medication has a statistically significantly higher average score in ASRS than unmedicated individuals. 6 individuals are taking medication to this day.The data collected confirm our main hypothesis that ADHD symptomatology has an impact on the daily functioning of individuals in adulthood. Unlike peers with reduced or no ADHD symptoms, these people are far more likely to have time management issues, need to plan their day carefully, and yet often experience problems such as late arrivals due to a lack of anticipation.Financial support was provided by grants : GA ČR – 18 -112 47 S, Progres Q06 1LF


To explore and compare the cognitive processing weakness of children with Attention deficit hyperactivity disorder (ADHD) and comorbid reading disabilities (RD) (ADHD+RD) and children with ADHD only using the Das-Naglieri Cognitive Assessment System (DN:CAS).n = 30) and ADHD group (n = 58) as well as the corresponding subgroups. Clinical data on gender, age, grade, IQ scores, and DN:CAS processing scores were compared between both groups/subgroups. Spearman's correlation test was used for correlation analysis of results of interest.Eighty-eight children with ADHD who visited the hospital for the first time from September 2021 to November 2021 and had a Full scale intelligence quotient (IQ) of ≥85 on the Wechsler Intelligence Scale for Children revised in China (C-WISC) were selected . Based on comorbidity with RD and the subtypes of ADHD , these children were divided into the ADHD+RD group (P > 0.05). Children in the ADHD-C+RD subgroup had lower scores in Planning processing of DN:CAS than those in the ADHD-C subgroup (P = 0.040). However, there were no significant difference between the ADHD-I+RD subgroup and ADHD-I subgroup in Planning scores of DN:CAS assessment; The grade of ADHD-C+RD and ADHD-I+RD subgroups were positively correlated with the Planning scores of DN: CAS . The grade of ADHD-C subgroup was positively correlated with the Planning and Simultaneous processing scores of DN: CAS .No differences in age, grade, male-to-female ratio, verbal IQ, performance IQ, and full scale IQ were observed between the ADHD+RD group and ADHD group as well as the corresponding subgroups (Our study confirmed that children of ADHD-C with comorbid RD have a more severe Planning processing weakness compared to children with ADHD-C only. Among the children of ADHD-C+RD, ADHD-I+RD and ADHD-C, such a Planning processing impairment may improve with increasing educational skills. Attention deficit hyperactivity disorder (ADHD) is a neurodevelopmental disorder characterized by attention deficit, hyperactivity, impulsivity, and cognitive dysfunction. According to the different clinical manifestations, it can be divided into Inattention dominant type (ADHD-I), Hyperactivity/Impulse dominant type (ADHD-H) and Combined type (ADHD-C) , 2. It oDas and Naglieri developed the PASS theory of intelligence in the 1990s based on the view of functional regions of brain operation proposed by A.R. Luria and study of cognitive psychology . The PASThere were many previous studies in children with ADHD, RD, and ADHD+RD by using DN:CAS. Most of the studies found that children with ADHD demonstrated relatively low scores on the Planning and Attention scales of the DN:CAS, but average scores on the Simultaneous and Successive scales –17. SomeIn our previous study, we found that the Verbal IQ, Performance IQ, and Full scale IQ scores of C-WISC of children with ADHD and comorbid SLD were significantly lower than those of children with ADHD only . In geneA total of 58 children with ADHD and 30 children with ADHD+RD were recruited in this study. All the children were enrolled from the Developmental Behavioral Pediatrics Clinic of the First Hospital of Jilin University from September 2021 to November 2021. ADHD was diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) criteria. Detailed clinical information should be collected for the diagnosis of ADHD, and the Vanderbilt ADHD scales , 23 inc. ChildreThe VARS was developed by Mark Wolraich of the University of Oklahoma according to the diagnostic criteria for ADHD in the DSM-IV. The toolkit is suitable for children aged 6–12 years. It consists of two subscales: Vanderbilt ADHD Diagnostic Parent Rating Scale (VADPRS) and Vanderbilt ADHD Diagnostic Teacher Rating Scale (VADTRS). Both subscales include behavior and performance assessment. For VADPRS, the section on behavior includes 47 items, which were divided into attention deficit (items 1–9), hyperactivity/impulsivity (items 10–18), oppositional defiance (items 19–26), conduct disorder (items 27–40), and anxiety/depression (items 41–47). For the VADTRS, the section on behavior includes 35 items, which are divided into attention deficit (items 1–9), hyperactivity/impulsivity (items 10–18), oppositional defiance (items 19–28), and anxiety/depression (items 29–35). Each item was graded at four levels: never (0), occasionally (1), often (2), and always (3). The performance section of both VADPRS and VADTRS contains eight items, which can quickly assess children's learning ability and interpersonal communication. Each item is rated as excellent (1), above average (2), average (3), somewhat problematic (4), or problematic (5). In 1998 and 2003, Professor Wolraich and other scholars conducted psychometric analysis on VADTRS and VADPRS, which showed that the scale had good reliability and validity , 23.WISC  was usedPRS is widely used to screen learning disabilities in China. It comprises the following five functional areas of verbal and non-verbal types: auditory comprehension and memory, language, time and orientation judgment, movement, and social behavior. The questionnaire has good reliability (retest correlation coefficients over 0.80) and fair validity  . The heaDCCC  applies DN:CAS is an intelligence test that operationalizes the PASS theory. The assessment system was suitable for individuals aged 5–17 years and included four subscales: Planning, Attention, Simultaneous processing, and Successive processing.Planning is involved in the cognitive process of executive function . The plan is modified as needed to stay consistent with the original goal. Planning assessment involves matching numbers and planned codes and planned connections.Attention processing includes expressive attention, number detection, and receptive attention. It involves focusing on cognitive activities, detecting specific stimuli, and inhibiting responses to distractive stimuli.Simultaneous processing includes non-verbal matrices, verbal spatial relations, and figure memory. This assessment requires the child to perceive the relationship between the components of the item and to integrate the separate elements into a complete pattern or concept of interrelation using verbal or non-verbal content.Successive processing assessment includes three tasks—word series, sentence repetition, and sentence questioning—which require individuals to understand and grasp information presented in a specific order.In this study, the Chinese version of the DN:CAS , which ht-test was used for comparison between groups. For categorical data, χ2 test was used for analyzing the difference between groups. The correlations among the DN:CAS scores and grade were detected using the Spearman's correlation test. P-value <0.05 was considered statistically significant.SPSS 22.0 statistical software  was used for all data analyses. Student's Detailed demographic data, IQ scores of C-WISC and the proportion of each ADHD subtype are available in n = 1; ADHD-H, n = 2) included in this study, it was not statistically analyzed.Due to the small number of ADHD-H subtypes   .P = 0.040)  .P > 0.05)  .P > 0.05)  .P = 0.030 and r = 0.508, P = 0.044, respectively). The grade of ADHD-C group was positively correlated with the Planning and Simultaneous processing scores of DN: CAS   .n = 1) and ADHD-H (n = 2) subgroups, the three children were not included in the statistics. No statistical differences in age, sex ratio, grade (years of schooling), Verbal IQ, Performance IQ and Full scale IQ scores were observed in all comparison. DN:CAS assessment was used to investigate the cognitive processing characteristics in ADHD+RD and ADHD children, and the differences between groups. The results show that ADHD-C+RD had lower scores in Planning processing when compared to ADHD-C. However, we found no significant differences between ADHD-I+RD and ADHD-I in all subscales of DN:CAS. Interestingly, years of schooling of children in ADHD-C+RD and ADHD-I+RD were positively correlated with their Planning processing of DN:CAS. In addition, the years of schooling were positively correlated with Planning and Simultaneous processing scores of DN: CAS in children with ADHD-C. Such correlations were not found in the ADHD-I.Eighty eight ADHD children with equal or <85 points of Full scale IQ according to the C-WISC were enrolled in our study. They were divided into six subgroups which are ADHD-C+RD, ADHD-C, ADHD-I+RD, ADHD-I, ADHD-H+RD and ADHD-H according to the presence of comorbid RD and the subtypes of ADHD. Due to the small number of ADHD-H+RD  as the point of interest and conducted correlation analysis with the cognitive processing of each subgroup. The Planning scores of ADHD-C+RD, ADHD-I+RD and ADHD-C children were positively correlated with their grade, whereas no such correlation was present in the ADHD-I subgroup. Some studies found that the relationship between cognitive processing and reading increased with age. However, no unified answer explains why the relationship is increased. One possible hypothesis is that the skills required for reading become more complex with age, increasing the involvement of the four PASS processes , 34. We This study had several limitations. First, since the Chinese language has pictographic characters and the English language has phonetic characters, we could not determine whether the cognitive processing characteristics of Chinese children with RD are similar to those of English children with RD. This limitation may weaken the comparability between the results of this study and previous studies to some extent. Second, due to the lack of a completely healthy control group, we could not determine the cognitive gap between children with ADHD+RD and their healthy peers. Therefore, our results can only be limited to the ADHD group. Third, this study excluded children with a Full scale IQ <85 on the C-WISC. Although the diagnoses of both RD and ADHD require adequate intelligence, Full scale IQ between 70 and 85 points is allowed. Moreover, quite a number of children with RD have a Full scale IQ <85. The results of this study could not represent the cognitive processing ability of all children with ADHD+RD. Subsequently, we will investigate whether children with lower Wechsler intelligence have different cognitive processing characteristics.Our study confirmed that children of ADHD-C+RD have more severe Planning processing weakness compared to children with ADHD-C. In contrast, there were no obvious differences between ADHD-I+RD and ADHD-I. But such impairment may improve with increasing educational skills among the children of ADHD-C+RD, ADHD-I+RD, and ADHD-C. And this conjecture should be identified and assessed with stronger evidence.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Ethical Committee of Jilin University First Hospital. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin. Written informed consent was obtained from the individual(s), and minor(s)' legal guardian/next of kin, for the publication of any potentially identifiable images or data included in this article.ZZ acquired the clinical data, reviewed the literature, and drafted the article. TW designed the study, supervised the initial drafting, and critically revised the article. JF, YX, and FJ analyzed the clinical data and critically revised the article. All authors approved the final version of the article.This work was supported by the National Natural Science Foundation of China (Grant Number: 81973054), Key Scientific and Technological Projects of Guangdong Province (Grant Number: 2018B030335001), and the Joint Fund Bethune Medical Special Project of Jilin Province (Grant Number: 20200201507JC).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Children diagnosed with attention deficit hyperactivity disorder (ADHD) require early intervention and sustained treatment. This study used a game-based digital treatment planning NeuroWorld DTx to analyze the impact on attention and cognitive function in children with ADHD. Thirty children diagnosed with ADHD were recruited and subjected to a four-week NeuroWorld DTx digital treatment. To assess the impact of the digital therapeutic therapy on children’s attention, we used the comprehension attention test (CAT) and Korean ADHD Rating Scale (K-ARS). Clinical global impression (CGI) and the Korean-child behavior checklist (K-CBCL 6–18) were used to examine the degree of improvement in ADHD. After four weeks, significant differences in the sensitivity and response style indices were noted, as compared with the baseline in the CAT test; in the case of K-ARS and CGI, a moderate decrease in ADHD was confirmed. The study achieved better results for the “total behavior problems” belonging to the K-CBCL assessment. Game-based digital therapy intervention can be a treatment method that elicits interest and satisfaction in children with ADHD and can be used as an adjunct to drug therapy to improve the quality of life and strengthen attention in children with symptoms of ADHD. The COVID-19 pandemic has, among its many direct and indirect effects on society, led to a mental health crisis among students . PsychiaThe symptomatic treatment of ADHD is currently based on drug treatment, which significantly improves the core symptoms ,12. Howep > 0.05) and sex  of the two groups were not significantly different. Detailed information regarding the study participants is shown in Thirty children aged six to thirteen years participated in this study. The number of study participants was calculated using the software G*power (version 3.1.2) ,22,23, wThe experimental group received NeuroWorld DTx and ADHD medication treatments, whereas the control group received the ADHD medication treatment alone. This helped verify the effect of NeuroWorld DTx. The two groups were designed before and after the experiment, as shown in This study was conducted from 19 January 2022 to 15 April 2022 at the Department of Mental Health Medicine, Keimyung University, Dongsan Hospital. In consideration of the ethical content of the subject, approval from the Institutional Bioethics Review Committee was obtained prior to data collection and evaluation. The study was performed after receiving approval (IRB No. 2021-10-080). The research participation agreement and description included information regarding the research purpose, participation procedure, participation risks and benefits, confidentiality, and not using the collected data for purposes other than research. After obtaining informed consent from both the participants and their guardians, the data collection and experiments were initiated. These data were kept obligatory, in accordance with the Bioethics Act.Attention was measured using the comprehension attention test (CAT), developed to evaluate various comprehensive types of attention and task execution functions for children and adolescents with ADHD ,25. The The degree of ADHD was measured using the attention deficit hyperactivity disorder rating scale, based on the DSM-IV ADHD diagnostic criteria ,30. The The overall clinical impression scale was used to evaluate the outcome of the worsening or improvement of symptoms before and after treatment ,33. ThisA tool standardized by K-CBCL 6–18, developed to evaluate the level of problem behaviors and social adaptation of children, was used for the parents of children and adolescents aged 6–18 years ,37. ThisIn this study, a tablet PC installed with NeuroWorld DTx, an AI-based attention and working memory improvement training program, was provided to each child participant. NeuroWorld DTx is a game-based program designed to enable home participation and therapy for attention deficit and impulsivity. NeuroWorld DTx is a game-based cognitive therapy software developed by Author and head of research center in Woorisoft Co., Ltd.,  of J.H.S. It is designed to enable home participation and therapy as an adjunct treatment for ADHD such as attention deficit and impulsivity. This study uses an unverified digital therapeutic agent is currently in the R&D stage. Therefore, considering that the participants in the study were vulnerable children between the ages of 6 and 13, the necessity and procedures of the program were explained to the guardians of the children and their prior consent was obtained. It can be used as an adjunct treatment for ADHD. NeuroWorld DTx explores the possibility of digital therapy as a mobile-based functional game for children with ADHD and evaluates whether digital therapy can be expected to improve attention and reduce ADHD symptoms. Notably, NeuroWorld DTx controls the excessive use of digital media to prevent side effects by automatically restricting the usage time to 30 min a day. NeuroWorld DTx comprises six items, four sets of attentiveness and two sets of working memory, as shown in p < 0.05 using SPSS for Windows 23.0 . The mean and standard deviation of the basic data were calculated and analyzed using a paired t-test to verify the homogeneity of the baselines for the two groups. In addition, the test results obtained in week 4 for the experimental and control groups were then compared with the baseline. The analysis of covariance (ANCOVA) was performed on the CAT and CGI scales. Korean ADHD Rating Scale (K-ARS) and Korean-Child Behavior Checklist (K-CBCL) were measured using the repeated measures ANOVA. The homogeneity test for the general characteristics of the experimental and control groups was conducted using Pearson and Fisher’s exact tests. Further, the t-test was employed as the homogeneity test for the dependent variable, when the normal distribution was satisfied, whereas the Mann–Whitney U test was used when the normal distribution was not satisfied [The statistical processing of data in this study was analyzed at a significance level of atisfied ,40.The NeuroWorld content satisfaction survey, which was used by both participants and parents as a training program, was conducted through a direct questionnaire. The percentages were calculated and analyzed. The efficacy evaluation was analyzed using modified intention-to-treat (ITT) and per-protocol (PP) analysis; however, in principle, the final decision of the study results was based on the modified ITT analysis results.The modified ITT analysis group was assigned to the NeuroWorld DTx group after randomization, and data collection on the primary efficacy endpoint was possible after baseline. Among the modified ITT analysis participants, those without significant violations  were used for the PP analysis under the study protocol procedure. The last observation carried forward (LOCF) method was used; this approach analyzes the data measured at the last visit by replacing the data measured at the last visit with the corresponding time point if dropouts occurred at a certain point in time for the efficacy endpoint. The safety evaluation involved checking whether all the data were coded and stored on an external hard drive separate from the computer for all the participants enrolled in this study and those who received NeuroWorld DTx at least once. To protect the confidentiality of the data, the personally identifiable information of the collected data was coded, the collected data were not used for purposes other than research, and the survey data were crushed and discarded after the data retention period, according to the Bioethics Act .p = 0.164), auditory selective attention task ; Flanker task , and sustained attention to response task . The attention task response style index includes the visual selective attention task , auditory selective attention task , Flanker task , and the sustained attention to response task . After four weeks of program intervention, Attention Task_Sensitivity Factor was 2 or higher in both the experimental and control groups in Visual Selective Attention Task, Auditory Selective Attention Task, and Sustained Attention to Response Task except for Flanker Task, and for Auditory Selective Attention Task, four weeks compared to baseline in the experimental group Afterwards, there was a significant increase , although no significant difference between the experimental and control groups was observed. Among the Attention Task_Sensitivity Factors, only the experimental group was significant in the Auditory Selective Attention Task . There was no statistically significant difference between the groups in the ANCOVA analysis with baseline correction in the Attention Task_Sensitivity Factor change rate. The Attention task_Response Style Index showed a significant difference (p = 0.020) between the experimental (0.70 ± 0.25) and control (0.57 ± 0.28) groups owing to the baseline-corrected ANCOVA in the Flanker task. In the Response Style Index change rate, the number of cases in which the Flanker Task index worsened was 14 (93.3%) in the control group compared to 8 (53.3%) in the experimental group. There was a statistically significant difference (p = 0.020) between the experimental group (−1.70 ± 29.14) and the control group (−25.43 ± 23.42) in the Flanker Task’s Response Style Index change rate. Detailed results are presented in In the CAT, the attention task sensitivity factor and attention task response style index were analyzed. The attention task sensitivity factor includes the visual selective attention task , K-ARS-Attention Deficit , and K-ARS-Hyper Activity  are presented herein. After four weeks of intervention, the total K-ARS score, Attention Deficit, and Hyper Activity of the experimental group were 12.27 ± 8.39, 7.27 ± 3.63, and 5.00 ± 5.11, respectively, which showed a statistically significant decrease compared to baseline . However, they were not significant in the control group . For additional details, please refer to For the ADHD rating scale, the total score, inattention, and hyperimpulsivity scores were evaluated as the rate of change, as compared with the baseline, at two weeks and four weeks after the experiment. The results for K-ARS . Furthermore, significant difference between the two groups in the rate of change (p= 0.023) was observed. For the experimental group, the measured severity of the symptoms  was significantly lower after four weeks of intervention . The control CGI-S score increased considerably but was not significant . The difference between the two groups was also significant . The degree of symptom improvement (CGI-I) was significantly lower in the experimental group (3.60 ± 0.63) than in the control group (4.07 ± 0.46) (p = 0.029). For additional details, please refer to The clinical global impression (CGI) was used to evaluate the degree of worsening or improvement of the symptoms before and after treatment. The severity of symptoms (CGI-S) was significantly lower in the experimental group (3.67 ± 0.62) than in the control group (4.53 ± 0.64) after four weeks of intervention (p = 0.963), K-CBCL_Internalizing Disorder , and K-CBCL_Externalizing Disorders  are presented herein. Statistically significant difference between the experimental group (15.66 ± 10.12) and the control group (6.68 ± 7.20) was observed in the Total Behavior Problems score change rate (p = 0.009). The experimental group showed a significant decrease  in the K-CBCL total problem behavior score. By contrast, the control group’s K-CBCL total problem behavior score increased significantly . There was no significant difference between the two groups . As a result of measuring the internalization score in the internalization and externalization rate of change, the experimental group showed a significant decrease . The control group had an increased score , and there was no difference between the two groups . In addition, although the externalization score decreased significantly for the experimental group , the control group showed a significantly increased score . There was no difference between the two groups .For the K-CBCL problem behavior and social adaptation, the scores and change rates of total problem behavior, internalization, and externalization in K-CBCL were compared . The resAfter applying NeuroWorld DTx, a satisfaction survey was conducted among the children and parents . The chiThis study investigated the effects of digital therapy on the attention function and symptoms of children diagnosed with ADHD. Clinical efficacy and safety were evaluated prospectively, on a single-center and standard-of-care basis. When combined with drug therapy, NeuroWorld DTx treatment led to an improved treatment effect in children with ADHD, as compared with the baseline. In this study, game-based digitization using NeuroWorld DTx was used. In the treatment regimen, the results of the four tests could summarize the attention of the study participants. CAT was analyzed using two indicators: sensitivity coefficient and response style. There was no significant difference in the scores between the experimental and control groups. However, four weeks after auditory simple selection attention, the sensitivity value of the experimental group showed a significant increase compared with the baseline, and the interference selection attention showed improvements in terms of the response style index. In the case of the attention deficit hyperactivity disorder (K-ARS), there was no significant difference in the K-ARS score and rate of change at two weeks, as compared with the baseline. However, after four weeks, a significant decrease in the degree of attention deficit hyperactivity disorder was confirmed in the experimental group, relative to the baseline. Significant effects in terms of the degree of severity and improvement (CGI) were also confirmed when NeuroWorld DTx and drug treatment were combined. In the case of the K-CBCL results, there were significant differences in the total scores for problem behavior, internalization, and externalization in the experimental group after four weeks of contrast intervention, as compared with the baseline. These results confirm that NeuroWorld DTx relieves the problem behavior syndrome. The improvement in ADHD symptoms after NeuroWorld DTx treatment achieved in this study was related to the existing digital therapy intervention, which is also valid for a previous case .Previous digital therapy intervention studies have shown that digital intervention can be applied to standard care without additional safety concerns and can help reduce the barriers to access, which are inherent in other forms of behavioral or non-pharmacological interventions . In partChildren with ADHD symptoms are usually preschoolers or young children. They exhibit a lack of motivation to solve problems, and their language is not fully developed. Thus, they are highly dependent on the environment and objects ,48. AccoTo prevent the negative effects of continuously using smartphones and digital media, the learning method selected in this study stipulated a usage time. This control is effective for preventing the abuse of functional game contents. Active intervention is considered to control the safety of these tools .A typical cognitive training program using digital media in the treatment process for children with ADHD symptoms is Endeavor Rx, which is a game software developed in the United States for the ADHD treatment of children aged 8–12 years . EndeavoNevertheless, this study has certain limitations. First, the participants included 30 children diagnosed with ADHD; here, the ADHD severity was considered as heterogeneous because the study was conducted on both patients who were previously diagnosed with ADHD and those who were diagnosed with ADHD for the first time. Second, only the short-term effects of digital therapy were analyzed. Although the analyses in this study afforded statistically significant results, observational studies over longer periods are required to show clinically significant results. Therefore, owing to the small number of participants and the short study period, there are limitations to various experimental factors. Hence, future studies considering the various long-term effects of digital therapy are required. Third, in this study, the age of children participating in the experiment was designated as between 6 and 13 years. Generally, 6 to 12 years of age is classified as childhood in the field of pediatric psychiatry, and those aged 13 to 18 are classified as adolescent. Therefore, in pediatric research, in general, the distribution of subjects for children’s studies is set from 6 to 12 years old, and the age range of participants in this study is also set from 6 to less than 13 years old, so the variables according to age change are not shown in detail. Therefore, in future, it is necessary to subdivide the age group and conduct a study to analyze the effectiveness according to age. Despite these limitations, NeuroWorld, which was used as an adjuvant therapy for drug treatment, showed a significant effect when combined with standard treatment. In addition, both the children and parents were satisfied with the NeuroWorld DTx digital therapy intervention and expressed their intention to re-participate. These findings suggest that the NeuroWorld Digital Therapy Program induces positive behavioral changes in children with ADHD. This also indicates its potential for use as an alternative treatment approach. In particular, it can be used as a therapeutic agent because of its excellent accessibility to children who miss the treatment period, despite the need for continuous treatment, owing to the spread of the COVID-19 pandemic. Therefore, in the future, digital adjuvant therapy and drug therapy can be used in parallel to improve the quality of life of children with ADHD.This study investigated the effect of a digital adjuvant therapy, NeuroWorld DTx, on the attention and working memory of children diagnosed with ADHD, as well as possibility of combining this approach with conventional drug treatment. Based on the CAT change rate, it was confirmed that the participants of the experimental group showed improved sensitivity and response style indexes, as compared with the control group. The rate of change in the K-ARS total score, inattention, and hyperimpulsivity scores significantly decreased for the experimental group after four weeks. In addition, when NeuroWorld DTx and the standard treatment were combined, a significant decrease in the CGI-S severity evaluation was noted, thereby suggesting the effectiveness of NeuroWorld DTx. NeuroWorld DTx was found to be effective as a digital therapy adjuvant therapy along with drug treatment; thus, it shows potential for use as a digital adjuvant therapy. Current research indicates that drug therapy is the optimal treatment for children with ADHD. However, convenient and effective treatment requires the development of alternative methods such as NeuroWorld DTx. Future research should focus on controlled trials with a large number of children, which will help evaluate NeuroWorld DTx’s effects on a variety of children with ADHD symptoms.


Enset is an important source of food and is consumed by about 25 million people as a staple or co-staple food crop mainly in southern parts of Ethiopia. Large numbers of enset landraces exist in different administrative zones of Ethiopia with a wide range of altitudes and agroclimatic zones. However, limited information is available on the diversity, distribution, and utilization pattern corresponding to the diverse ethnolinguistic as well as sociocultural communities of the country. Hence, this study was devised to explore and document the richness of farmers’ tradition and practice on the diversity and distribution of enset landraces on the farm level and selection pattern for different purposes regarding the production, utilization, and conservation of enset genetic resources.The study was conducted in four major enset-growing administrative zones of Ethiopia, namely Hadiya, Kembata-Tembaro, Gurage, and Silte. A total of 240 farm households were surveyed using individual interviews, 18 key informant interviews, 36 focus group discussions with 5 participants, and direct on-farm field observations for data collection. Considering that enset has a rich cultural background and indigenous knowledge, ethnobotanical research approach was applied to data collection and analysis. The Shannon–Weaver, Simpson, Pielou, and Jaccard’s similarity indices were used to evaluate the diversity and similarity of the landraces as well as using descriptive statistics in SPSS Ver. 24. Preference in direct matrix ranking was also used to compute and rank the enset landraces most preferred by the people in the context of specific use value in the study area.A total of 282 farmer-named enset landraces have been identified, with a range from 2 to 32 on individual homegardens. The largest number of landraces was found in the Hadiya Zone (86), while the lowest was scored in the Silte Zone (57). The Shannon diversity index (H') ranged from 3.73 (Silte) to 3.96 (Hadiya). Similarly, landraces revealed a very narrow range of variances in Simpson’s 1-D diversity index, and it ranged from 0.963 (Silte) to 0.978 (Hadiya). Likewise, the similarity index ranged from 0.24 to 0.73 sharing 16–47 landraces in common. Of the 282 landraces, 210 (74.5%) were recorded in more than one zones, whereas 72 (25.5%) had narrow distribution being restricted to a single zone.Farmers have established long-term practices and experiences in cultivation, utilization, and conservation of a diverse group of enset landraces to fill their domestic and market purposes in each zone. The variation is likely to be related to agroclimatic differences, ethnicity factors, food cultures, and historical backgrounds. Therefore, to facilitate on-farm conservation as well as sustainable utilization of the enset genetic resources, farmers need to be supported by different stakeholders for all their worth and also in crop improvement programs. Ensete ventricosum (Welw.) Cheesman] is a large perennial monocarpic herbaceous plant, similar to the banana in form, in the family Musaceae within the monocot order of Zingiberales [E. ventricosum is domesticated, and the corm (short underground stem) and pseudostem (thick and soft midrib) are processed and consumed as a staple and co-staple food in the south and southwestern parts of Ethiopia [Enset [iberales . E. ventEthiopia . Enset iEthiopia . It is nEthiopia , 5. MoreEthiopia , 6, 7 wiEthiopia . This diEthiopia –11.qocho, the juice from the scrapings known as bulla, and the boiled corm known as amicho), rate of maturation, disease and drought tolerance, forage quality, medicinal value, ease of scraping, quality of corm, and productivity [Farmers have managed the diversity of enset landraces for centuries with limited or no research influences from outside , 13 beinuctivity , 20, 21.Understanding the diversity and distribution of enset is crucial for sustainably managing genetic resources and crop improvement efforts. Yemataw and co-workers  showed tShigeta  describeqocho, bulla, and amicho. Furthermore, some enset varieties are used traditionally to cure bone fractures, birth problems, and diarrhea in humans [The high genetic diversity of enset warrants conservation, as it provides resilience to the enset farming system and thus food security for farming communities , 16, 18.n humans , 32.Enset landraces are grown in homegardens with different local names and often with wide distribution and varietal diversity with implications to genetic diversity. For the sustainable utilization and on-farm conservation of its genetic resources as well as future improvement of the crop, understanding the sociocultural, ethnobotanical knowledge, farmers’ selection criteria, and retention practices of enset landrace diversity in different ethnolinguistic communities is vital. However, limited documentation . The study woredas and kebeles were selected from each administrative zone based on enset diversity where prior information was obtained from the departments of agriculture of the respective zones and woredas  were selected purposefully based on enset frequency of occurrence and production level. From each woreda, three kebeles were also chosen purposefully according to the information obtained from the agricultural office of each woreda. Therefore, a total of 36 kebeles were selected for data collection. From each kebele, six to seven individual farmers were selected randomly that make a total of 240 households (60 household heads from each zone) in the whole study sites.For this study, a multistage sampling method was performed for the selection of individual enset-growing farmers in each zone. Of the four administrative zones, 12 woreda official was informed of the study's objectives and wrote supporting letters to notify their respective kebele administrative offices. After obtaining the kebele leaders' permission, the investigator, local elders, and the agricultural extension workers of each kebele had a comprehensive discussion about the study's objectives and a schedule for the fieldwork and interview sessions. Following verbal informed consent of each informant, interviews and discussions were conducted to gather indigenous knowledge  held by knowledgeable informed volunteers and participants about the on-farm diversity, use patterns, and traditional management practice of the enset crop.The Microbial, Cellular, and Molecular Biology Department (Addis Ababa University) initially reviewed the study proposal. Following the approval, a supporting letter was written to the zonal administrative offices of the study area adhering to the existing national guidelines. As a result, each district/Both primary and secondary data collection methods were conducted to assess and document farmers’ local knowledge regarding on-farm diversity, distribution, and utilization of enset crops in the study area. Two rounds of data collection and field observation were conducted .kebeles in local languages  and in the Amharic official language. The principal investigator can also communicate and understand three of the above-listed local languages; hence this made our work easier and the communication very smooth.To develop semi-structured interviews, different kinds of discussion were conducted initially with three to four elder enset farmers in each zone to generate needed information to be collected in the study area. In-depth individual interviews were conducted together with trained agricultural extension workers, who are working closely with the communities in the respective selected woreda agricultural experts, agricultural extension workers of kebeles, elderly farmers, and local leaders. For the focus group discussions, from each of the selected woredas, about five participants were involved together with the members of the local administration, community elders, agricultural extension workers, and other members of participating communities. Additional data were also collected through, preference in direct matrix ranking by involving 12 key informants (three from each zone). Secondary data were also reviewed from the reports of the agricultural office of each zone, different books, research articles, and journals.The farmers were also asked about their perception of names and naming systems. To obtain the detailed local knowledge of farmers in each zone, 4–5 key informants were also selected based on prior information obtained from woreda (district) with the help of knowledgeable senior farmers. Moreover, some minor dialect variations in naming landraces within the same ethnic group were not considered different and were disregarded in landrace authentication. However, landraces having the same names, but originating from different ethnic groups or zones, were documented as its. The collected survey data were analyzed using descriptive statistics  in SPSS Ver. 24. The landrace richness, distribution, and abundance per homegarden were also calculated using Microsoft Excel 2010. Richness was computed to show the total number of landraces per homegarden based on data recorded in each administrative zone as this is a simple applicable biodiversity index to use and compare diversity in enset landraces. Abundance was determined as the total number of individual enset plants of each landrace per homegarden. Preference in direct matrix ranking was conducted to analyze the most preferred enset landraces, in the context of the four specific use values for the seven enset landraces. Twelve key informants participated in the arrangement of the values by giving the most favored enset landraces a score of 10, the least preferred enset a score of 1, 0 for the uses not known, and the others a score that fell somewhere in between. Based on the total scores obtained for each landrace, these values were then summed for all respondents and ranked.All listed landraces throughout the collection sites were checked for known synonyms or local names that refer to the same or different landraces in each study zone and ith landrace.Diversity and similarity indices of species can be quantified in different ways. In this study, the diversity indices were calculated from the number of landraces existing in 60 farmers’ homegardens within each zone. The Shannon and Weaver  and SimpEven though Shannon’s index takes into account the evenness of the abundance of landraces, evenness (equitability) can also be computed separately. It is a measure of the proportion of the observed diversity for the maximum diversity expected and was calculated through the Pielou index  as the rD) is a measure of diversity. It measures the probability that two individuals randomly selected from an area will belong to the same species [Simpson’s diversity index ( species  and hencD) = 1 − {Σn (n − 1)/N (N − 1)} The value of this index ranges between 0 and 1; the greater the value, the greater the diversity, 1 represents infinite diversity and 0, no diversity. The index was computed for all study zones.b) diversity [a is the number of landraces at zone A, b is the number of landraces at zone B, and J is the number of landraces common to both locations. Sorenson's similarity coefficient ranges in value from 0 (no similarity) to 1 (complete similarity).Sorenson similarity index was employed to assess differentiation or beta  and internal features (leaf and midrib anatomy and fiber structure) of the enset plants, and they give distinct vernacular names for each landrace. Each local farmer in the studied area was observed cultivating a diverse of enset landraces in his or her homegarden, which shows a considerable variation in the number of enset landraces on individual homegardens. It ranges from two to thirty-two in this study Table . In comp.  describe. [. , who des. [.  and Yema. [.  stated tThe Shannon diversity index (H′) ranged from 3.73 (Silte) to 3.96 (Hadiya), this signifies the existence of a high richness of enset landraces in the study zones. Even though zones varied in richness, they revealed a very narrow range of variances in Simpson’s 1-D and evenness indices. The Simpson’s 1-D ranged from 0.963 (Silte) to 0.978 (Hadiya) and evenness indices ranged from 0.89 to 0.92. All these results specify the presence of high enset landraces diversity in these four zones (Table The similarity among pairs of zones (taking two zones at a time) concerning farmers-named landraces was evaluated using Sorenson’s similarity index Table . GeneralAgade, Astara, Bededete/Badade, Gimbo/Gimbuwa, Heniwa/Hiniba/Enba, Kasete, Manduluqa/Mande, Mariye, Merza, Mesmesia, Moche, Separa/Sebera, Torora/Xorore, Weshemeja and Zobira  were widely distributed in all four zones. These were ra Table . Similarra Table . Likewisra Table . But theGimbo, Hiniba, and Separa were relatively high in abundance at all four study zones. Agade, Bedededa, and Zobira were also other most frequent enset landraces in three out of the four zones  and its different growth stages vary from one ethnic group to another. Enset is called wessa in Hadiya and Kembata-Tembaro, wesse in Silte, and aset in Gurage. Moreover, each growth (transplanting) stage has a distinct name by which it is identified. The Hadiya and Kembata-Tembaro farmers share almost the same local names for all sucker stages. These are known as dubbo, simma, ero/kiniba, and balwesa, but in Silte 1 -and 2-year-old suckers are called bosho and daporo, respectively, and the other two stages are nearly similar to the Hadiya and Kembata-Tembaro zones  when parts are cut. The origin of certainly cultivated enset is evident from the name. One such example in this study is Kembat which may be originated from Kembata; however, its name in Kembata-Tembaro and Hadiya is called Disho , but this landrace in Hadiya is given the name Meqelwesa, meaning placental enset, which is related to use characters. Similarly, landraces Soqido is salt (taste of boiled corm or amicho) in Hadiya while in Silte and Gurage it is Kemele meaning Ape (maybe the color of the pseudostem or petiole)   Table . In genenstance, , 15, andnstance,  reportednstance, , 13, andnstance,  stated tnstance, , banana nstance, , sweet pnstance, , cassavanstance, , and comnstance, , 43. Oure et al.  and Abera) Table . MoreoveMeqelwesa or Qeqile-qey which was rarely used as food. This landrace is one of the most traditionally preferred medicinal enset landraces recommended for human and cattle ailments  tasty and medicinal value, and Sisqella, Bededede, and Gimbo stood first to third, respectively for their fiber quality  were identified as having narrow distribution in the study zones. However, in some cases the same kinds of enset are known with alternative local names used as medicines for different problems among the study communities  quoted by many farmers to remove the delayed placenta and for aborification (used to cause/facilitate abortion) purposes  production except landraces Gishira, Dare, and Bedededa. In the same manner, the most chosen part of enset for medicinal use was corm but the landrace Meqelwesa (in Hadiya) or Qeqile-qey (in Kembata) were all part used as traditional medicine. In some cases, farmers also used cooked qocho or porridge prepared from bulla to treat different health problems in the study zones  for duplicates identification and clarification of synonymies, and to facilitate their on-farm conservation as well as sustainable utilization of enset farming communities and also in its improvement programs. A new study shows that frequent severe drought events led to an increase in enset production areas in Ethiopia. Indigenous staples are “saviors” during difficult times. This is why national investment in their conservation, improvement, and value addition is necessary for a changing climate.


There was limited evidence to suggest a causal effect of genetic liability to ADHD  or ASD  on Alzheimer’s disease. Similar causal effect estimates were identified as direct effects, independent of educational attainment  and IQ . Genetic liability to Alzheimer’s disease was not found to have a causal effect on risk of ADHD or ASD . We found limited evidence to suggest a causal effect of genetic liability to ADHD or ASD on Alzheimer’s disease; and vice versa.Few studies suggest possible links between attention deficit hyperactivity disorder (ADHD), autism spectrum disorder (ASD) and Alzheimer’s disease but they have been limited by small sample sizes, diagnostic and recall bias. We used two-sample Mendelian randomization (MR) to estimate the bidirectional causal association between genetic liability to ADHD and ASD on Alzheimer’s disease. In addition, we estimated the causal effects independently of educational attainment and IQ, through multivariable Mendelian randomization (MVMR). We employed genetic variants associated with ADHD , ASD , Alzheimer’s disease , educational attainment ( Attention deficit hyperactivity disorder (ADHD) and autism spectrum disorder (ASD) are lifelong neurodevelopmental conditions associated with large societal costs –3. It haADHD is characterised by difficulties in several areas of neurocognitive functioning, including memory, attention and inhibitory control . In lateIn the case of ASD, it has been suggested that abnormal synaptogenesis, connectivity, and hyper plasticity of the brain identified recently in ASD, could be protective against Alzheimer’s disease . Adding Beyond these hypotheses, little is currently known on the possible associations between ADHD, ASD and Alzheimer’s. There is only a small number of case-control studies on ADHD, suggesting a higher frequency of ADHD symptoms in patients with dementia with Lewy body and a higher risk of dementia in adults with ADHD –17. HoweThe release of the latest ADHD and ASD GWASs provides a unique opportunity to investigate the possible causal associations between genetic liability to ADHD, ASD and Alzheimer’s, through a method that overcomes limitations of observational studies: Mendelian randomization. Mendelian randomization is an instrumental variables (IVs) approach using genetic variants as proxies for exposures to investigate the causal effects of these exposures on health outcomes . As geneFor these reasons, we aimed to investigate the bidirectional causal associations between genetic liability to ADHD, ASD and Alzheimer’s using two-sample Mendelian randomization. Next, we performed multivariable Mendelian randomization to estimate the direct causal effects of genetic liability to ADHD and ASD on risk of Alzheimer’s, independent of educational attainment and IQ to explore the possibility that the causal effect of ADHD and ASD on Alzheimer’s are being masked by IQ or educational attainment.A.must be strongly associated with the exposure,B.independently of any confounders of the exposure-outcome association, andC.are associated with the outcome only via their effect on the exposure .MR relies on strict assumptions that the genetic variants should satisfy in order to be considered valid instruments and therefore yield unbiased causal effect estimates. Specifically, the genetic variants:In the context of the present study, we applied two-sample MR , a methoWe used GWAS summary data from the latest publicly available GWASs on the exposures of interest. Table r2 < 0.01 within a 10,000 kb window, p < 5 × 10−08) and corresponding log odds ratios and standard errors were extracted from the publicly available datasets. An exception was ASD, where only two independent variants were identified. Therefore, in order to increase the power of our study, we relaxed the significance threshold (p < 5 × 10−07) for instrument selection. A similar approach for ASD instrument selection has been used in a previous study [r2 < 0.01 within a 10,000 kb window).For each exposure of interest independent genetic variants were identified  method. IVW can be used to summarize the causal effects of multiple genetic variants, as it is equivalent to fitting a weighted linear regression of the gene-outcome associations on the gene-exposure associations, with the intercept term constrained to zero , 29. ThuIn order to explore the validity of MR assumptions, we compared the estimated total causal effects obtained from univariable MR to the effects obtained using the MR-Egger regression and the Weight median estimator , 31. Bri2), the number of genetic variants used and the sample size. An F-statistic smaller than 10 indicates the presence of weak instrument bias and that causal effect estimates are likely to be influenced.In addition, as the validity of MR results depends largely on the strength of the genetic instruments, we used F-statistic to ensure that weak instrument bias will not affect our results . F-statiThe influence of each genetic variant on the outcome, was assessed via a leave-one out analysis, in which genetic variants are systematically removed and causal effects of the remaining SNPs on the outcome are re-estimated . FinallyMVMR is an extension of MR that can be utilized in cases that multiple exposures seem to be strongly genetically related and are considered to have possible causal effects on an outcome. MVMR allows the estimation of the direct effects of each exposure on the outcome by entering the exposures within the same model. Detailed information on the method can be found in the original publication .The main analyses conducted in the present study used summary data from Phase 3 of the Alzheimer’s GWAS, which included proxy-cases of Alzheimer’s disease in UK Biobank Table . This miConsidering the above limitations, we conducted a second round of analyses using summary data of the meta-analysis of Korologou- Linden et al.  which coAll statistical analysis was performed in R 3.6.1, TwoSampleMR and MendelianRandomization packages were used for the harmonization of datasets and estimation of causal effects. All statistical tests were two-sided.Instrument strength as estimated by the F-statistic did not indicate weak instrument bias as it ranged from 30 to 51 for ADHD, 25 to 35 for ASD and 30 to 945 for Alzheimer’s instruments.P = 0.39) on Alzheimer’s disease. Horizontal pleiotropy was unlikely to bias the result, as suggested by the MR-Egger intercept term . Moreover, MR-Egger and weighted median estimators yielded similar results to the IVW estimator for ADHD , and leave-one out analyses did not identify any SNP as influential . MR-Egger and Weight median estimators were directionally consistent with the IVW , and leave-one out analysis did not point out any genetic variant as influential , a total of 643 and of 180 variants were available for inclusion in the ADHD-EA-AD and ADHD-IQ-AD multivariable analyses, respectively.P = 0.76) (Table P = 0.29)  Table . A simil9) Table .Table 3DThe results were comparable—in univariable and multivariable MR analyses—to the ones obtained from the MR analyses using Phase 1 of the Alzheimer’s disease GWAS which did not include proxy cases (Supplementary Tables P = 0.70) on Alzheimer’s disease. MR-Egger and weighted median estimators produced directionally comparable results to the IVW estimator (Table P = 0.58). No considerable heterogeneity was observed between the genetic variants  and leave-one out analyses did not identify any SNP as influential . MR-Egger and weighted median estimators were directionally consistent with the IVW for the genetic liability of Alzheimer’s on ASD (Table P = 0.53) and by Cochrane’s Q statistic . Moreover, leave-one out analysis did not point out any genetic variant as influential  (Table P = 0.99)  Table . In addi9) Table .Table 5DThe results were comparable to the ones obtained from the MR analyses using Phase 1 of the Alzheimer’s disease GWAS which did not include proxy cases .Investigating the possible associations of ADHD and ASD with Alzheimer’s disease is an important area of research that can have important implications for the affected individuals, their families and policy makers. It is important therefore, for future GWAS studies to offer subtype specific data in order to explore any possible differential associations of ADHD and ASD subtypes with Alzheimer’s. In addition, although longitudinal designs for this question could be difficult, the availability of registry data  can offer a unique opportunity to investigate the associations between ADHD, ASD and Alzheimer’s disease as well as the possible role of educational attainment through their extensive clinical and academic records.This is the first study to investigate the possible causal associations between genetic liability to ADHD, ASD and Alzheimer’s disease using the largest sample sizes publicly available for each phenotype, within a Mendelian randomization framework. We found limited evidence to suggest total and direct effects of genetic liability to ADHD and ASD on risk of Alzheimer’s disease. We hope that this will be an important step towards encouraging future research into possible differential associations of ADHD and ASD subtypes and risk of Alzheimer’s disease as well as utilizing longitudinal data.Supplementary material


Rye is the only cross-pollinating small-grain cereal. The unique reproduction biology results in an exceptional complexity concerning genetic improvement of rye by breeding. Rye is a close relative of wheat and has a strong adaptation potential that refers to its mating system, making this overlooked cereal readily adjustable to a changing environment. Rye breeding addresses the emerging challenges of food security associated with climate change. The systematic identification, management, and use of its valuable natural diversity became a feasible option in outbreeding rye only following the establishment of hybrid breeding late in the 20th century. In this article, we review the most recent technological advances to improve yield and yield stability in winter rye. Based on recently released reference genome sequences, SMART breeding approaches are described to counterbalance undesired linkage drag effects of major restorer genes on grain yield. We present the development of gibberellin-sensitive semidwarf hybrids as a novel plant breeding innovation based on an approach that is different from current methods of increasing productivity in rye and wheat. Breeding of new rye cultivars with improved performance and resilience is indispensable for a renaissance of this healthy minor cereal as a homogeneous commodity with cultural relevance in Europe that allows for comparatively smooth but substantial complementation of wheat with rye-based diets, supporting the necessary restoration of the balance between human action and nature. Secale cereale L.) entered its European growth habitat as a weedy grass in wheat/barley agricultural systems and evolved as a secondary crop by initially mimicking the visual appearance of Triticum spp. and Hordeum vulgare [Rye , the staple crop for an estimated 35% of the human population, which will continue to play a critical role to ensure an adequate and affordable intake of calories and proteins in diets until 2050 [T. aestivum approximately 3–4 million years ago [In recent decades, European agriculture has experienced higher, more pronounced and more widespread climate variability as compared to the long-term average . In 2022til 2050 . Rye divears ago  and is cears ago ,13. The ears ago . In factears ago . These rears ago  to a chaThe systematic identification, management, and use of valuable natural diversity in outbreeding rye only became a feasible option following the establishment of hybrid breeding late in the 20th century. The rationale and main driver for hybrid breeding is the systematic exploitation of economic or standard heterosis that refers to the superiority of a hybrid over standard commercial check varieties. In rye, hybrid breeding is a success story in terms of the selection of favorable alleles for grain yield. In sharp contrast to the competitiveness of hybrid compared to line breeding in wheat , the keyper se performance and is designed to measure alterations in highly heritable traits such as plant height, thousand-grain weight, and disease resistance with high sensitivity  systems ,46,47,48Both commercially used CMS systems differ in their impact on hybrid rye breeding. The G cytoplasm belongs to a plasmotype that is common in the Central European rye gene pool . Its invRf) genes. Major Rf genes enabling grain production in CMS-based hybrids are available for both the G and the P cytoplasm, and have been mapped on the distal end of the long arm of chromosome 4R [Rfp genes originating from non-adapted genetic resources is mainly monogenic dominant-inherited [Rf genes on the short arm of chromosome 1RS and the long arm of chromosome 3R [In hybrid progenies from selective mating, the CMS effect of the seed-parent genotype needs to be counteracted by nuclear alleles from the pollen parent that restore pollen production. Such genetic factors, thus, are designated restorer-of-fertility  Tulasne, the species of greatest concern [Ergot is a disease of cereals and grasses caused by fungi in the genus  concern . At anthC. purpurea virulence factors have been reported, RNAseq analysis of in planta-expressed fungal genes identified more than 400 highly expressed transcripts including an elevated frequency of genes encoding putative effectors that might be involved in repelling the fungal attack or interfering with host-defense interactions [Rf genes is the most effective approach for minimizing ergot infestation in hybrid rye. Because of the toxicity of ergot alkaloids for humans and animals, the European Commission has significantly lowered the thresholds for ergot since 1 January 2022. Supplementary to the previous threshold of 0.05% ergot sclerotia in unprocessed rye, harvested grain may not contain more than 500 µg/kg of total ergot alkaloids in rye milling products for human consumption [Noteworthy in this context is a protective effect provided by ergot on cereal rye that probably acted against human exploitation, as heavily affected seed and grain samples have been, and still are, disposed of. This example of conditional host-pathogen interaction in which the consequence may fall on different parts of a continuum from parasitism to mutualism depending on the specific situation might explain why grasses are commonly susceptible and rather tolerant to ergot and no effective resistance mechanisms have evolved over time . Indeed,ractions . Althougractions , specifisumption . From 1 Rfp1 only [Rfp1 originates from a weedy rye population from the Fertile Crescent [Rfp1 in rye have been reported influencing seed development through male gametogenesis or post-fertilization, due to factors deposited by the sperm cells in the egg and/or central cells, as well as by genomic imprinting [Arabidopsis thaliana, with a much larger influence of the paternal genome on F1 seed size than previously appreciated [1 seed size has been observed in this study as well. These results from fundamental research have implications for breeding research on the seed biology of hybrid rye and open new avenues for genetic approaches to compensate for paternal contributions to seed size in rye hybrids with strong ergot defense. Susceptibility to ergot upon artificial inoculation is a trait assessed for registered rye varieties since 2008 in the descriptive variety list of the German Federal Plant Variety Office. The description of rye varieties listed in 2022 for their resistance and yield traits demonstrates genetic gain for ergot defense in highly productive hybrids already at the level of open-pollinating cultivars . Howeverfp1 only , the devCrescent ,65,70,78Crescent . This reCrescent . Paternaprinting . Recentlreciated . NotablyRfp genes while simultaneously minimising the costs of restoration in terms of grain yield. Particularly promising for this purpose are genes that cause parent-of-origin effects on F1 seed size such as MET1, the primary maintenance DNA methyltransferase in Arabidopsis thaliana. Crosses between Arabidopsis genotypes carrying a complete loss-of-function and the wild-type MET1 allele revealed that hypomethylated maternal genomes result in significantly larger F1 seeds [SECCE2Rv1G0093370 [MET1. In total, 6 SNP markers on the rye 600k SNP-array [SECCE2Rv1G0093370 between elite inbred lines and germplasm collections. Subsequently, an ingenious cross-validation strategy [SECCE2Rv1G0093370. Superior SNP alleles leading to hypomethylated maternal genomes as measured by significantly larger F1 seeds are attractive means to offset the undesired paternal inherited effects of Rfp1.Our steadily growing knowledge of the genetic code of grain size and weight control ,82 and kF1 seeds . In rye,G0093370  shares 5NP-array  provide This drafted option for selection with markers and advanced reproductive technology (SMART) breeding  in rye eRfp1 on grain yield but may result in insufficient quantities of pollen to combat the fungus adequately in farmers’ fields, as wet pollen agglutinates and distributes over short distances only. In fact, multi-environmental data on natural ergot infection provided by the Polish Research Centre for Cultivar Testing (COBORU) confirm this assumption  of them become available. The current breeding scheme is two generations shorter than the standard scheme [per se performance are selected from the base population and crossed to two heterozygous testers representing the ‘Carsten’ and ‘Petkus’ pool. The testcrosses are subsequently evaluated in 3–4 target environments with respect to their yield performance to identify the GCA of the selected genotypes  . RRS is  testers . A greatd scheme , and thuenotypes D. Based 2 are used to establish further testcrosses to achieve robust performance data on yield potential, stability, and adaptation with the second GCA test in 8 to 10 highly divergent environments in terms of soil fertility and precipitation . Recently published reference genome sequences ,42 and hKnowledge of genomic regions controlling complex traits is the key to our understanding of mechanisms behind yield architecture and to using them in marker-assisted crop improvement programs. The novel catalog of SNP variations associated with yield and grain weight holds potential for the clear-cut and systematic genetic improvement of yield potential in rye hybrids with strong ergot defense. Hybrid breeding offers a pragmatic strategy to evaluate the influence of single genomic loci on grain yield in rye. Progenies segregating for superior (S) or inferior (I) SNP alleles can be used to establish near-isogenic genotype bulks (NIB) of homozygous elite F3 lines of the seed-parent pool, that will serve as pollinators in crosses with male-sterile single-cross testers from the opposite pool between isolation walls . In the Although the development of perfect markers that address the functional sequence polymorphism of a gene governing grain yield would be the consistent final step, it needs to be considered that the necessary functional characterization of a candidate gene by transposon tagging, transformation for overexpression, knockdown analysis, or genetic modification based on novel plant breeding techniques is still in its infancy, i.e. not available or published in rye. The extension of the molecular toolbox available for rye by sophisticated technologies probably is a long-term perspective. In contrast, the proposed International Functional Genomics Project on Rye (IFGPR) for the development of highly replicable rye genetic stocks in the public domain  appears Dominant dwarf 1 (Ddw1), identified five decades ago by Vladimir Kobylyansky in the germplasm collection at the N.I. Vavilov Institute of Plant Genetic Resources [Estestvennyj Mutant 1 , has been used in breeding of open pollinating cultivars for the genetic improvement of plant height [Ddw1 has not been used in hybrid breeding programs due to theoretical considerations [The introgression of major dwarfing genes for increasing yield potential is a proven concept in wheat and rice ,130,131.esources  and initt height , a quantt height ,134,135.t height ,138,139.t height ,140,141 erations ,134,135.erations . Ddw1 in hybrid rye, novel markers [Ddw1 into the seed-parent pool. Homozygous semidwarf BC3S2 single-cross testers enabled the development of semidwarf P-type hybrids in 2020 for the first time ever. Subsequently conducted performance trials in European and Canadian target environments for rye cultivation revealed that the semidwarfs were distinct from existing conventional hybrid rye varieties and expressed the genetically reduced plant height uniformly , the semidwarfs revealed a pronounced lodging resistance as a result of their shorter and sturdier stems . In viewproducts . SemidwaDdw1 belongs to the group of gibberellin (GA)-sensitive dwarfing genes [Eragrostis tef (Zucc.) Trotter) and finger millet (Eleusine coracana Gaertn) have been reported as a result of chemically induced GA deficiency [GA2ox2/6 and an increased drought tolerance compared to the wild-type plants. In semidwarf rye the gene ScGA2ox12 co-segregating with the GA-sensitive dwarfing gene Ddw1 is up-regulated as well [Ddw1 mutant alters the GA content in a favourable manner. Notably, the integration of the Ddw1 target interval in the wheat reference genome sequence indicated perfect micro-colinearity between the Ddw1 locus and a 831 kb segment on chromosome 5A, which resides inside of a 11.21 Mb interval carrying the GA-sensitive dwarfing gene Rht12 in wheat [Rht12 overgrowth mutants with a range of intermediate height phenotypes taller than the original Rht12 stock [ng genes . Noteworng genes . Positivficiency . In riceficiency . In accoficiency . The ect as well , supportin wheat . The kno12 stock  as novelDdw1 on the line per se performance in thousand-grain weight and flowering time have been reported [Ddw1 on the testcross performance of genetically divers rye hybrids, and in a reasonable number of environments, were lacking until recently. Indeed, extensive phenotyping of conventional experimental hybrids representing a high level of genetic diversity in two years with contrasting precipitation patterns improved our understanding of rye’s adaptation to drought stress [Ddw1 as an option to tap currently unused potential for optimized dry matter allocation to the rye grain. However, it may be misleading to transfer knowledge from conventional to semidwarf genotypes without empirical data. For instance, in semidwarf Ddw1 genotypes the head has proven to be supplied with assimilates essentially via the flag leaf (F) and F-1 [Ddw1 on the performance of rye in target environments of rye cultivation. The enhanced implementation of Ddw1 in hybrid rye breeding serves as a viable example of breeders’ options to target allelic combinations of genes controlling phenology and plant architecture. In this way, further known major genes such as VRN, PPD, EPS [Concerns have been raised about a potentially negative impact of genetically reduced culm length in rye on photosynthesis and yield performance ,135,142.reported . Howevert stress . This co and F-1 , in cont and F-1 . The comPPD, EPS , and genPPD, EPS  are wortDdw1 in a practical hybrid rye breeding program is paradigmatic for specific genes and adaptive alleles that govern important agronomic traits in rye. The promising performance in target environments of rye cultivation triggers an enhanced development of semidwarf hybrids and may initiate a new era of physiological rye breeding that aims to raise the yield potential of rye closer to its biological limit. Indeed, the rich genetic diversity of rye, together with a sophisticated bulked segregant phenotyping strategy of testcross performance in multi-environmental field trials, argue for a stronger utilization of rye in research directed to the identification of valuable alleles controlling yield and yield components for Triticeae improvement programs. For this purpose, further investment in resequencing is promising for an improved understanding of the genetics of adaptive traits in rye.Despite current achievements, major challenges in rye production remain for advancing rye from its current status as a healthy minor cereal to a trend-setting crop with increasing relevance for grain production in a changing climate. Therefore, improved breeding efforts are strategically important for enhancing the competitiveness of rye in agricultural production systems. In rye, hybrid breeding serves as a cutting edge technology to select crossing partners on a rational basis for breeding improved cultivars with different complex but complementary traits to achieve cumulative gene action for grain yield and yield stability. The identification of traits that may allow rye to capture its most limiting resources more efficiently receives strong support from two high-quality rye genome assemblies ,42 and pHybrid breeding had obvious advantages and positive side effects in rye. In contrast to many valuable crops with a relatively small acreage that became orphans for which breeding was no longer carried out , hybrid