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Melaku Tadege,1 Azmeraw Misganaw,2 Zemenay Truneh,1 Awoke Seyoum Tegegne3 1Department of Statistics, Injibara University, Injibara, Amhara, Ethiopia; 2Department of Statistics, Mettu University, Mettu, Oromia, Ethiopia; 3Department of Bio-Statistics, Bahir Dar University, Bahir Dar, EthiopiaCorrespondence: Awoke Seyoum Tegegne, Email bisrategebrail@yahoo.comIntroduction: Currently, diabetes is a global health problem and it affects many people, especially in the developing continents. As patients’ living conditions improve and the science of medicine advances, the longevity of such patients has increased greatly. Therefore, the purpose of this study was to identify predictors for the association of the longevity of people with diabetes in Buno Bedele and Illubabor Zones, South-west Ethiopia.Methods: The study applied a retrospective cohort study design approach. In particular, long rank tests for longevity experience and Cox semi-parametric regression were implemented to compare and investigate the predictors associated with the longevity of patients with diabetes.Results: Among all the patients who participated in this study, 56.9% were females and the rest were males. From the Cox regression result, age (AHR = 1.0550, 95% CI: (1.0250, 1.0860), p-value = 0.001), female patients (AHR = 0.2200, 95% CI: (0.0390, 0.5290)), rural patients (AHR = 0.2200, 95% CI: (0.1000, 0.4890), p-value = 0.001), the existence of fasting blood glucose complication (AHR = 1.2040, 95% CI: (1.0930, 1.4460), p-value = 0.001), the existence of blood pressure (AHR = 1.2480, 95% CI: (1.1390, 1.5999), p-value = 0.0180), treatment type, Sulfonylureas (AHR = 4.9970, 95% CI: (1.4140, 17.6550), p-value = 0.0120), treatment type, Sulfonylurea and Metformin (AHR = 5.7200, 95% CI: (1.7780, 18.3990), p-value = 0.0030) were significantly affected the longevity of people with diabetes.Conclusion: The findings of the current study showed that the patient’s age, sex of patients, residence area, the existence of complications, existence of pressure, and treatment type were major risk factors related to the longevity of people with diabetes. Hence, health-related education should be given to patients who come to take treatment to have better longevity for people with diabetes. More attention should be given to aged patients, male and urban patients, patients under complication treatment, and patients under treatment with single-treatment medication.Keywords: DMD, mortality, Log rank, Cox proportional hazards model, longevity

Hiwot Yisak,1 Melaku Tadege,1 Birhanie Ambaw,1 Amien Ewunetei2 1Department of Public Health, College of Health Sciences, Debre Tabor University, Debre Tabor, Ethiopia; 2Department of Pharmacy, College of Health Sciences, Debre Tabor University, Debre Tabor, EthiopiaCorrespondence: Hiwot Yisak Tel +251-913728949Email hyisak@yahoo.comBackground: Underweight, wasting, and stunting are the commonest nutritional disorders among children, especially in developing countries. The aim of this study was to assess the prevalence and determinant factors of underweight, wasting, and stunting among school-age children in 2019.Methods: A cross-sectional study was conducted in the five special districts of South Gondar Zone, among 314 school-age children. WHO AnthroPlus software was used to build Z-scores from anthropometric measurement. The data were analyzed by SPSS Version 20. The degrees of association were assessed using adjusted odds ratio (AOR) and 95% confidence interval during multivariable logistic regression. A P-value less than 0.05 was considered to be statistically significant.Results: Of the total study participants, 232 (77.3%) were from public schools. The mean±standard deviation (SD) of height of children was 132.9± 9.8 cm, and the mean±SD weight of children was 27.7± 5.8 kg. The prevalence of stunting, wasting, and underweight was 11%, 6.3%, and 11.4%, respectively. Students who ate their breakfast rarely were 8-times more likely to be underweight than those who ate their breakfast always (AOR=7.9, 95% CI=4.8– 14.8). Those who were sick in the past 2 weeks were more likely to be underweight than their counterparts (AOR=7.3, 95% CI=2.8– 14.4). Those who never consume milk or milk products were 6.5 (AOR=6.5, 95% CI=1.7– 23) times more likely to be stunted than those who consumed this always. Sickness in the past 2 weeks prior to data collection was significantly associated with thinness (AOR=6 0.9, 95% CI=4.1– 10.1).Conclusion: The overall prevalence of wasting, stunting, and underweight was a mild public health problem in the study area.Keywords: wasting, stunting, underweight, school-age children, South Gondar, Ethiopia

Desalegn Tesfa,1 Wallelign Aleminew,2 Melaku Tadege,1 Alemayehu Digssie,1 Bedilu Abebe,1 Sofonyas Abebaw,1 Belayneh Kefale3 1Department of Public Health, College of Health Sciences, Debre Tabor University, Debre Tabor, Ethiopia; 2Department of Health Education, University of Gondar, Gondar, Ethiopia; 3Department of Pharmacy, College of Health Sciences, Debre Tabor University, Debre Tabor, EthiopiaCorrespondence: Desalegn Tesfa Department of Public HealthCollege of Health Sciences, Debre Tabor University, P.O. Box 031, Debre Tabor, EthiopiaTel +251-918027124Email desalegntesfa50@gmail.comBackground: Happiness is the most indicator of good quality of life particularly for pregnant women. Reduced happiness of pregnant women can result in reduced maternal health services utilization and can result in adverse newborn and maternal outcomes. Assessing the level of happiness and its factors are important for pregnant women and the next generation. Hence, the current research paper aimed to assess the level of happiness and its associated factors among pregnant women who attended antenatal care follow-up in South Gondar Zone Hospitals.Methods: Institution-based cross-sectional study was carried out from September 2018 to February 2019 among 1606 expectant pregnant women in the last 6 months. The Oxford Happiness Questionnaire (OHQ) was used as a measure of the degree of happiness and used four as average person score. Besides, bivariate, multivariable logistic analyses were deployed to identify associations.Results: A total of 1606 pregnant women were participating in this study. The prevalence of low level of pregnancy happiness in this study was 24% (95% CI: 20.7 − 26.6). Mothers who exposed to unintended pregnancy (AOR; 1.94, CI 95% 1.86– 4.52), reported intimate partner violence (AOR; 2.42, 95% CI 1.67– 3.54) and being in the third trimester (AOR; 1.89 95% CI 1.19– 3.01) were the most predictor variables with the low level of happiness.Conclusion: Approximately one-fourth of the pregnant women had a low level of pregnancy happiness. Mothers who were exposed to unintended pregnancy encountered intimate partner violence, who were in the third trimester were the most significant variables associated with a low level of happiness. Governmental and non-governmental organizations work on women’s reproduction rights and the zonal health department with local health offices should educate on the prevention of intimate partner violence and unintended pregnancy and its consequences and also empower women to cope with changes and challenges of pregnancy and childbirth.Keywords: happiness, pregnancy, women, level, associated factors

Multimedia content delivery via the cellular infrastructure increases fast due to the very high volumes of mobile video traffic generated by the billions of end devices populating the mobile data network. A critical mass of mobile video content requests refers to the consumption of the same popular video content, which is consumed by different end terminals spanning small geographical regions. Such content requests place a great burden on the backhaul of content-agnostic cellular networks, which fail to exploit the correlation of video requests to decongest their backhaul links. This creates redundant retransmissions while fetching the same video content from a central server to the network edge, using the bandwidth-limited backhaul at peak-time periods. With the integration of multi-access edge computing (MEC) capabilities in 5G mobile cellular networks, mobile network operators can place popular video content closer to the network edge at off-peak time periods, predicting user requests exhibiting a high correlation for a given time interval over smaller geographical regions. In this paper, we investigate popular content placement in multi-tier heterogeneous cellular networks where the edge network infrastructure can cooperate to create content delivery (and placement) clusters to effectively serve correlated video requests. To this end, we model the cooperative content placement problem using the multiple knapsack problem (MKP) formulation and present an exact (optimal) bound-and-bound strategy to solve it. The performance of the proposed strategy is evaluated in-depth using extensive system-level simulations and is compared against that of other state-of-the-art algorithms. Valuable design guidelines and key performance trade-offs are discussed, paving the way towards cluster-based cooperative caching in MEC-enabled cellular network setups.

Introduction: Cardiovascular diseases are a class of heart and blood vessel-related illnesses. In Sub-Saharan Africa, including Ethiopia, preventable heart disease continues to be a significant factor, contrasting with its presence in developed nations. Therefore, the objective of the study was to assess the prevalence of death due to cardiac disease and its risk factors among heart patients in Ethiopia., Methods: The current investigation included all cardiac patients who had cardiac surgery in the country between 2012 and 2023. A total of 1520 individuals were participated in the study. Data collection took place between February 2022 and January 2023. The study design was a retrospective cohort since the study track back patients' chart since 2012. Machine learning algorithms were applied for data analysis. For machine learning algorithms comparison, lift and AUC was applied., Results: From all possible algorithms, logistic algorithm at 90%/10% was the best fit since it produces the maximum AUC value. In addition, based on the lift value of 3.33, it can be concluded that the logistic regression algorithm was performing well and providing substantial improvement over random selection. From the logistic regression machine learning algorithms, age, saturated oxygen, ejection fraction, duration of cardiac center stays after surgery, waiting time to surgery, hemoglobin, and creatinine were significant predictors of death., Conclusion: Some of the predictors for the death of cardiac disease patients are identified as such special attention should be given to aged patients, for patients waiting for long periods of time to get surgery, lower saturated oxygen, higher creatinine value, lower ejection fraction and for patients with lower hemoglobin values., (© 2024. The Author(s).)

The milestone of compound leaf development is the generation of separate leaflet primordia during the early stages, which involves two linked but distinct morphogenetic events: leaflet initiation and boundary establishment for leaflet separation. Although some progress in understanding the regulatory pathways for each event have been made, it is unclear how they are intrinsically coordinated. Here, we identify the PINNATE-LIKE PENTAFOLIATA2 (PINNA2) gene encoding a newly identified GRAS transcription factor in Medicago truncatula. PINNA2 transcripts are preferentially detected at organ boundaries. Its loss-of-function mutations convert trifoliate leaves into a pinnate pentafoliate pattern. PINNA2 directly binds to the promoter region of the LEAFY orthologue SINGLE LEAFLET1 (SGL1), which encodes a key positive regulator of leaflet initiation, and downregulates its expression. Further analysis revealed that PINNA2 synergizes with two other repressors of SGL1 expression, the BEL1-like homeodomain protein PINNA1 and the C2H2 zinc finger protein PALMATE-LIKE PENTAFOLIATA1 (PALM1), to precisely define the spatiotemporal expression of SGL1 in compound leaf primordia, thereby maintaining a proper pattern of leaflet initiation. Moreover, we showed that the enriched expression of PINNA2 at the leaflet-to-leaflet boundaries is positively regulated by the boundary-specific gene MtNAM, which is essential for leaflet boundary formation. Together, these results unveil a pivotal role of the boundary-expressed transcription factor PINNA2 in regulating leaflet initiation, providing molecular insights into the coordination of intricate developmental processes underlying compound leaf pattern formation., Competing Interests: Conflict of interest statement. The PINNA2 gene investigated in this study has been the subject of an international Patent Cooperation Treaty (PCT) application with the patent number PCT/CN2023/072361. The authors affirm that they have no additional competing interests to declare., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

Molecular genetic understanding of flowering time regulation is crucial for sorghum development. GRAIN NUMBER, PLANT HEIGHT AND HEADING DATE 7 (SbGhd7) is one of the six classical loci conferring photoperiod sensitivity of sorghum flowering. However, its functions remain poorly studied. The molecular functions of SbGhd7 were characterized. The gene regulatory network controlled by SbGhd7 was constructed and validated. The biological roles of SbGhd7 and its major targets were studied. SbGhd7 overexpression (OE) completely prevented sorghum flowering. Additionally, we show that SbGhd7 is a major negative regulator of flowering, binding to the promoter motif TGAATG(A/T)(A/T/C) and repressing transcription of the major florigen FLOWERING LOCUS T 10 (SbFT10) and floral activators EARLY HEADING DATE (SbEhd1), FLAVIN-BINDING, KELCH REPEAT, F-BOX1 (SbFKF1) and EARLY FLOWERING 3 (SbELF3). Reinforcing the direct effect of SbGhd7, SbEhd1 OE activated the promoters of three functional florigens (SbFT1, SbFT8 and SbFT10), dramatically accelerating flowering. Our studies demonstrate that SbGhd7 is a major repressor of sorghum flowering by directly and indirectly targeting genes for flowering activation. The mechanism appears ancient. Our study extends the current model of floral transition regulation in sorghum and provides a framework for a comprehensive understanding of sorghum photoperiod response., (© 2024 The Authors New Phytologist © 2024 New Phytologist Foundation.)

Introduction: Living in poverty, especially in low-income countries, are more affected by cardiovascular disease. Unlike the developed countries, it remains a significant cause of preventable heart disease in the Sub-Saharan region, including Ethiopia. According to the Ethiopian Ministry of Health statement, around 40,000 cardiac patients have been waiting for surgery in Ethiopia since September 2020. There is insufficient information about long-term cardiac patients' post-survival after cardiac surgery in Ethiopia. Therefore, the main objective of the current study was to determine the long-term post-cardiac surgery patients' survival status in Ethiopia., Methods: All patients attended from 2012 to 2023 throughout the country were included in the current study. The total number of participants was 1520 heart disease patients. The data collection procedure was conducted from February 2022- January 2023. Machine learning algorithms were applied. Gompertz regression was used also for the multivariable analysis report., Results: From possible machine learning models, random survival forest were preferred. It emphasizes, the most important variable for clinical prediction was SPO2, Age, time to surgery waiting time, and creatinine value and it accounts, 42.55%, 25.17%,11.82%, and 12.19% respectively. From the Gompertz regression, lower saturated oxygen, higher age, lower ejection fraction, short period of cardiac center stays after surgery, prolonged waiting time to surgery, and creating value were statistically significant predictors of death outcome for post-cardiac surgery patients' survival in Ethiopia., Conclusion: Some of the risk factors for the death of post-cardiac surgery patients are identified in the current investigation. Particular attention should be given to patients with prolonged waiting times and aged patients. Since there were only two fully active cardiac centers in Ethiopia it is far from an adequate number of centers for more than 120 million population, therefore, the study highly recommended to increase the number of cardiac centers that serve as cardiac surgery in Ethiopia., (© 2024. The Author(s).)

Alfalfa ( Medicago sativa L.) forage quality is adversely affected by lignin deposition in cell walls at advanced maturity stages. Reducing lignin content through RNA interference or antisense approaches has been shown to improve alfalfa forage quality and digestibility. We employed a multiplex CRISPR/Cas9-mediated gene-editing system to reduce lignin content and alter lignin composition in alfalfa by targeting the COUMARATE 3-HYDROXYLASE (MsC3H) gene, which encodes a key enzyme in lignin biosynthesis. Four guide RNAs (gRNAs) targeting the first exon of MsC3H were designed and clustered into a tRNA-gRNA polycistronic system and introduced into tetraploid alfalfa via Agrobacterium -mediated transformation. Out of 130 transgenic lines, at least 73 lines were confirmed to contain gene-editing events in one or more alleles of MsC3H . Fifty-five lines were selected for lignin content/composition analysis. Amongst these lines, three independent tetra-allelic homozygous lines ( Msc3h-013, Msc3h-121 , and Msc3h-158 ) with different mutation events in MsC3H were characterized in detail. Homozygous mutation of MsC3H in these three lines significantly reduced the lignin content and altered lignin composition in stems. Moreover, these lines had significantly lower levels of acid detergent fiber and neutral detergent fiber as well as higher levels of total digestible nutrients, relative feed values, and in vitro true dry matter digestibility. Taken together, these results showed that CRISPR/Cas9-mediated editing of MsC3H successfully reduced shoot lignin content, improved digestibility, and nutritional values without sacrificing plant growth and biomass yield. These lines could be used in alfalfa breeding programs to generate elite transgene-free alfalfa cultivars with reduced lignin and improved forage quality., Competing Interests: 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. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Wolabu, Mahmood, Chen, Torres-Jerez, Udvardi, Tadege, Cong, Wang and Wen.)

Plant-specific WUSCHEL-related homeobox (WOX) family transcription factors play critical roles in maintaining meristems and lateral organ development. The WUS clade member STF/LAM1 physically interacts with the intermediate clade member WOX9. This interaction contributes to their antagonistical functions on leaf blade outgrowth by competing for the same cis-elements in the promoter of their common target in M. truncatula and N. sylvestris. Here, we identified the main interaction domains of STF and MtWOX9 in Medicago, shedding light on the mechanism of WOX gene function. The middle domain of STF and MtWOX9 are both critical for the interaction, while the conserved motif of STF in the C-terminal domain is also required. Deletion of the middle domain of STF partially rescued the leaf blade phenotypes of the stf null mutant, indicating that the middle domain plays an essential role during leaf blade expansion. This finding provides a new insight that the versatility of WOX function is not only caused by the conserved DNA binding and repression domains but also by the middle domain that recruits different partners., (© 2024 Scandinavian Plant Physiology Society.)

Plant lateral organs are often elaborated through repetitive formation of developmental units, which progress robustly in predetermined patterns along their axes. Leaflets in compound leaves provide an example of such units that are generated sequentially along the longitudinal axis, in species-specific patterns. In this context, we explored the molecular mechanisms underlying an acropetal mode of leaflet initiation in chickpea pinnate compound leaf patterning. By analyzing naturally occurring mutants multi-pinnate leaf1 (mpl1) that develop higher-ordered pinnate leaves with more than forty leaflets, we show that MPL1 encoding a C2H2-zinc finger protein sculpts a morphogenetic gradient along the proximodistal axis of the early leaf primordium, thereby conferring the acropetal leaflet formation. This is achieved by defining the spatiotemporal expression pattern of CaLEAFY, a key regulator of leaflet initiation, and also perhaps by modulating the auxin signaling pathway. Our work provides novel molecular insights into the sequential progression of leaflet formation., (© 2023. The Author(s).)

Plant organ size is an important agronomic trait tightly related to crop yield. However, the molecular mechanisms underlying organ size regulation remain largely unexplored in legumes. We previously characterized a key regulator F-box protein MINI ORGAN1 (MIO1)/SMALL LEAF AND BUSHY1 (SLB1), which controls plant organ size in the model legume Medicago truncatula. In order to further dissect the molecular mechanism, MIO1 was used as the bait to screen its interacting proteins from a yeast library. Subsequently, a KIX protein, designated MtKIX8, was identified from the candidate list. The interaction between MIO1 and MtKIX8 was confirmed further by Y2H, BiFC, split-luciferase complementation and pull-down assays. Phylogenetic analyses indicated that MtKIX8 is highly homologous to Arabidopsis KIX8, which negatively regulates organ size. Moreover, loss-of-function of MtKIX8 led to enlarged leaves and seeds, while ectopic expression of MtKIX8 in Arabidopsis resulted in decreased cotyledon area and seed weight. Quantitative reverse-transcription PCR and in situ hybridization showed that MtKIX8 is expressed in most developing organs. We also found that MtKIX8 serves as a crucial molecular adaptor, facilitating interactions with BIG SEEDS1 (BS1) and MtTOPLESS (MtTPL) proteins in M. truncatula. Overall, our results suggest that the MIO1-MtKIX8 module plays a significant and conserved role in the regulation of plant organ size. This module could be a good target for molecular breeding in legume crops and forages., (© 2023 Scandinavian Plant Physiology Society.)

Anthocyanin and catechin production in tea (Camellia sinensis) leaves can positively affect tea quality; however, their regulatory mechanisms are not fully understood. Here we report that, while the CsMYB75- or CsMYB86-directed MYB-bHLH-WD40 (MBW) complexes differentially activate anthocyanin or catechin biosynthesis in tea leaves, respectively, CsMYBL2a and CsMYBL2b homologs negatively modified the light- and temperature-induced anthocyanin and catechin production in both Arabidopsis and tea plants. The MBW complexes activated both anthocyanin synthesis genes and the downstream repressor genes CsMYBL2a and CsMYBL2b. Overexpression of CsMYBL2b, but not CsMYBL2a, repressed Arabidopsis leaf anthocyanin accumulation and seed coat proanthocyanin production. CsMYBL2b strongly and CsMYBL2a weakly repressed the activating effects of CsMYB75/CsMYB86 on CsDFR and CsANS, due to their different EAR and TLLLFR domains and interactions with CsTT8/CsGL3, interfering with the functions of activating MBW complexes. CsMYBL2b and CsMYBL2a in tea leaves play different roles in fine-tuning CsMYB75/CsMYB86-MBW activation of biosynthesis of anthocyanins and catechins, respectively. The CsbZIP1-CsmiR858a-CsMYBL2 module mediated the UV-B- or cold-activated CsMYB75/CsMYB86 regulation of anthocyanin/catechin biosynthesis by repressing CsMYBL2a and CsMYBL2b. Similarly, the CsCOP1-CsbZIP1-CsPIF3 module, and BR signaling as well, mediated the high temperature repression of anthocyanin and catechin biosynthesis through differentially upregulating CsMYBL2b and CsMYBL2a, respectively. The present study provides new insights into the complex regulatory networks in environmental stress-modified flavonoid production in tea plant leaves., (© 2023 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Alfalfa (Medicago sativa L.) is a perennial flowering plant in the legume family that is widely cultivated as a forage crop for its high yield, forage quality and related agricultural and economic benefits. Alfalfa is a photoperiod sensitive long-day (LD) plant that can accomplish its vegetative and reproductive phases in a short period of time. However, rapid flowering can compromise forage biomass yield and quality. Here, we attempted to delay flowering in alfalfa using multiplex CRISPR/Cas9-mediated mutagenesis of FLOWERING LOCUS Ta1 (MsFTa1), a key floral integrator and activator gene. Four guide RNAs (gRNAs) were designed and clustered in a polycistronic tRNA-gRNA system and introduced into alfalfa by Agrobacterium-mediated transformation. Ninety-six putative mutant lines were identified by gene sequencing and characterized for delayed flowering time and related desirable agronomic traits. Phenotype assessment of flowering time under LD conditions identified 22 independent mutant lines with delayed flowering compared to the control. Six independent Msfta1 lines containing mutations in all four copies of MsFTa1 accumulated significantly higher forage biomass yield, with increases of up to 78% in fresh weight and 76% in dry weight compared to controls. Depending on the harvesting schemes, many of these lines also had reduced lignin, acid detergent fibre (ADF) and neutral detergent fibre (NDF) content and significantly higher crude protein (CP) and mineral contents compared to control plants, especially in the stems. These CRISPR/Cas9-edited Msfta1 mutants could be introduced in alfalfa breeding programmes to generate elite transgene-free alfalfa cultivars with improved forage biomass yield and quality., (© 2023 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

The plant-specific family of WUSCHEL (WUS)-related homeobox (WOX) transcription factors is key regulators of embryogenesis, meristem maintenance, and lateral organ development in flowering plants. The modern/WUS clade transcriptional repressor STENOFOLIA/LAMINA1(LAM1), and the intermediate/WOX9 clade transcriptional activator MtWOX9/NsWOX9 antagonistically regulate leaf blade expansion, but the molecular mechanism is unknown. Using transcriptome profiling and biochemical methods, we determined that NsCKX3 is the common target of LAM1 and NsWOX9 in Nicotiana sylvestris. LAM1 and NsWOX9 directly recognize and bind to the same cis-elements in the NsCKX3 promoter to repress and activate its expression, respectively, thus controlling the levels of active cytokinins in vivo. Disruption of NsCKX3 in the lam1 background yielded a phenotype similar to the knockdown of NsWOX9 in lam1, while overexpressing NsCKX3 resulted in narrower and shorter lam1 leaf blades reminiscent of NsWOX9 overexpression in the lam1 mutant. Moreover, we established that LAM1 physically interacts with NsWOX9, and this interaction is required to regulate NsCKX3 transcription. Taken together, our results indicate that repressor and activator WOX members oppositely regulate a common downstream target to function in leaf blade outgrowth, offering a novel insight into the role of local cytokinins in balancing cell proliferation and differentiation during lateral organ development., (© American Society of Plant Biologists 2022. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Root nodule formation in many leguminous plants is known to be affected by endogen ous and exogenous factors that affect formation, development, and longevity of nodules in roots. Therefore, it is important to understand the role of the genes which are involved in the regulation of the nodulation signaling pathway. This study aimed to investigate the effect of terpenoids and terpene biosynthesis genes on root nodule formation in Glycine max . The study aimed to clarify not only the impact of over-expressing five terpene synthesis genes isolated from G. max and Salvia guaranitica on soybean nodulation signaling pathway, but also on the strigolactones pathway. The obtained results revealed that the over expression of GmFDPS , GmGGPPS , SgGPS , SgFPPS , and SgLINS genes enhanced the root nodule numbers, fresh weight of nodules, root, and root length. Moreover, the terpene content in the transgenic G. max hairy roots was estimated. The results explored that the monoterpenes, sesquiterpenes and diterpenes were significantly increased in transgenic soybean hairy roots in comparison with the control. Our results indicate the potential effects of terpenoids and terpene synthesis genes on soybean root growth and nodulation. The study provides novel insights for understanding the epistatic relationship between terpenoids, root development, and nodulation in soybean.

Tea (Camellia sinensis) is concocted from tea plant shoot tips that produce catechins, caffeine, theanine, and terpenoids, which collectively determine the rich flavors and health benefits of the infusion. However, little is known about the integrated regulation of shoot tip development and characteristic secondary metabolite biosynthesis in tea plants. Here, we demonstrate that MYB transcription factors (TFs) play key and yet diverse roles in regulating leaf and stem development, secondary metabolite biosynthesis, and environmental stress responses in tea plants. By integrating transcriptomic and metabolic profiling data in different tissues at a series of developmental stages or under various stress conditions, alongside biochemical and genetic analyses, we predicted the MYB TFs involved in regulating shoot development (CsMYB2, 98, 107, and 221), epidermal cell initiation (CsMYB184, 41, 139, and 219), stomatal initiation (CsMYB113 and 153), and the biosynthesis of flavonoids (including catechins, anthocyanins, and flavonols; CsMYB8 and 99), caffeine (CsMYB85 and 86), theanine (CsMYB9 and 49), carotenoids (CsMYB110), mono-/sesquiterpenoid volatiles (CsMYB68, 147, 148, and 193), lignin (CsMYB164 and 192), and indolic compounds (CsMYB139, 162, and 198), as well as the MYB TFs that are likely involved in hormone signaling-mediated environmental stress and defense responses. We characterized the functions of some key MYBs in regulating flavonoid and carotenoid biosynthesis for tea quality and flavor. This study provides a cross-family analysis of MYBs in tea alongside new insights into the coordinated regulation of tea plant shoot development and secondary metabolism, paving the way towards understanding of tea quality trait formation and genetic improvement of quality tea plants., (© 2022 Society for Experimental Biology and John Wiley & Sons Ltd.)

Nyctinastic leaf movement of Fabaceae is driven by the tiny motor organ pulvinus located at the base of the leaf or leaflet. Despite the increased understanding of the essential role of ELONGATED PETIOLULE1 ( ELP1 )/ PETIOLE LIKE PULVINUS ( PLP ) orthologs in determining pulvinus identity in legumes, key regulatory components and molecular mechanisms underlying this movement remain largely unclear. Here, we used WT pulvinus and the equivalent tissue in the elp1 mutant to carry out transcriptome and proteome experiments. The omics data indicated that there are multiple cell biological processes altered at the gene expression and protein abundance level during the pulvinus development. In addition, comparative analysis of different leaf tissues provided clues to illuminate the possible common primordium between pulvinus and petiole, as well as the function of ELP1. Furthermore, the auxin pathway, cell wall composition and chloroplast distribution were altered in elp1 mutants, verifying their important roles in pulvinus development. This study provides a comprehensive insight into the motor organ of the model legume Medicago truncatula and further supplies a rich dataset to facilitate the identification of novel players involved in nyctinastic movement.

Soil-transmitted helminthiasis (STHs) and undernutrition are common health problems in developing countries. Several reports showed that STH and undernutrition are often associated. The main aim of this study was to determine the association of STH and undernutrition among schoolchildren in Mettu town, Southwest Ethiopia. A cross-sectional study design was employed. To collect socio-demographic data, semi-structured questionnaire and physical observation were used. Kato-Katz technique and Anthropometric measurements were also considered to see STH infection and determine the nutritional statuses of the study participants respectively. Then, the data generated from the study was managed using Chi-square test and logistic regression analysis to determine the association of demographic variables with infections of helminthes and assess the risk factors for nutritional status of the study participants respectively. As a result, among the 392 study schoolchildren, 331 (84.4%) children were positive for different species of STH and undernutrition accounted 32.6%. Ascaris lumbricoides (39.0%), Trichuris trichiura (32.9%) and hookworm (28.1%) are the predominant STH identified from the study participants. Age, maternal educational and occupation status, and fingernail status of children were found significantly associated (p < 0.05) with the risk of getting STH. Bivariate logistic regression analysis showed that, age (AOR 2.18, 95% CI 1.53, 6.59), maternal illiteracy (AOR 0.13, 95% CI 0.91, 0.34) and maternal occupation (AOR 1.67, 95% CI 1.08, 5.91) were major co-founding factors for the prevalence of STH among study participants. In addition, children with T. trichiura infection were more likely (P < 0.01) to suffer from undernutrition (AOR 0.52, 95% CI 0.31, 0.83). Thus, the findings revealed the high prevalence of STH and it has significant association with undernutrition among school age children in the study area. Anti-helminthic mass drug administration and maternal health education should be anticipated to curve the tragedy., (© 2022. The Author(s).)

In legumes, many endogenous and environmental factors affect root nodule formation through several key genes, and the regulation details of the nodulation signaling pathway are yet to be fully understood. This study investigated the potential roles of terpenoids and terpene biosynthesis genes on root nodule formation in Glycine max . We characterized six terpenoid synthesis genes from Salvia officinalis by overexpressing SoTPS6, SoNEOD, SoLINS, SoSABS, SoGPS , and SoCINS in soybean hairy roots and evaluating root growth and nodulation, and the expression of strigolactone (SL) biosynthesis and early nodulation genes. Interestingly, overexpression of some of the terpenoid and terpene genes increased nodule numbers, nodule and root fresh weight, and root length, while others inhibited these phenotypes. These results suggest the potential effects of terpenoids and terpene synthesis genes on soybean root growth and nodulation. This study provides novel insights into epistatic interactions between terpenoids, root development, and nodulation in soybean root biology and open new avenues for soybean research., Competing Interests: 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., (Copyright © 2021 Ali, Miao, Hou, Darwish, Alrdahe, Ali, Benedito, Tadege, Wang and Zhao.)

Homeobox transcription factors are key regulators of morphogenesis and development in both animals and plants. In plants, the WUSCHEL-related homeobox (WOX) family of transcription factors function as central organizers of several developmental programs ranging from embryo patterning to meristematic stem-cell maintenance through transcriptional activation and repression mechanisms. The Medicago truncatula STENOFOLIA (STF) gene is a master regulator of leaf-blade lateral development. Here, the crystal structure of the homeodomain (HD) of STF (STF-HD) in complex with its promoter DNA is reported at 2.1 Å resolution. STF-HD binds DNA as a tetramer, enclosing nearly the entire bound DNA surface. The STF-HD tetramer is partially stabilized by docking of the C-terminal tail of one protomer onto a conserved hydrophobic surface on the head of another protomer in a head-to-tail manner. STF-HD specifically binds TGA motifs, although the promoter sequence also contains TAAT motifs. Helix α3 not only serves a canonical role as a base reader in the major groove, but also provides DNA binding in the minor groove through basic residues located at its C-terminus. The structural and functional data in planta reported here provide new insights into the DNA-binding mechanisms of plant-specific HDs from the WOX family of transcription factors., (open access.)

Background. Substances mainly khat, alcohol and cigarette are used during pregnancy in Ethiopia. However, to this date, there is no pooled evidence about the burden of adverse neonatal outcomes among the substance users during pregnancy in the country. Methods . Eligible primary studies were accessed from 4 international data bases (Google Scholar, Science Direct, Scopus, and PubMed). The required data were extracted from these studies and then exported to stata version 14 for analysis. Subgroup analyses were conducted for evidence of heterogeneity. Results . A total of 2298 neonates were included from 7 studies. Among these neonates, 530(23.06%) were those whose mothers used substance during pregnancy (exposed group) whereas 1768 neonates were those whose mothers didn't use substance during pregnancy (controls group). The pooled prevalence of adverse neonatal outcome among the exposed mothers was 38.32% (95% CI: 29.48%, 47.16%; I 2  = 76.3%) whereas it was 16.29% (95% CI: 9.45%, 23.13%) among the controls. Adverse neonatal outcome was most burdensome among cigarette smokers 45.20% (95% CI: 37.68%, 52.73%; I 2  = .00%) when compared with khat chewers 34.00% (95% CI: 20.87%, 47.13%) and alcohol drinkers 38.47% (95% CI: 17.96%, 58.98%). Low birth weight 42.00% (95% CI: 18.01%, 65.99%; I 2  = 91.8%) was the most common adverse birth outcome. Conclusion . It was found that adverse neonatal outcomes were much more burdensome among antenatal substance users than the controls. Therefore, mothers should be enabled to quit using substance before pregnancy. Besides, strict comprehensive screening of every pregnant mother should be made at antenatal care clinics for early identification and management of antenatal substance use., Competing Interests: Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2021.)

The growth of leaves and biosynthesis of characteristic secondary metabolites are critically important for tea production and quality control. However, little is known about the coordinated regulation of leaf development and catechin biosynthesis in tea plants. Here, we reported that TCP TFs are involved in both catechin biosynthesis and leaf development. An integrated analysis of catechin profiling and CsTCP expression in different tissues of plants under various environmental conditions at different developmental stages indicated significant correlations between the transcript levels of CIN-type TCPs and catechin production. CIN-type CsTCP3 and CsTCP4 and PCF-type CsTCP14 interacted with the MYB-bHLH-WD40 repeat (MBW) complex by forming a CsTCP3-CsTT8 heterodimer and modulating the transactivation activity of the promoters of anthocyanin synthase (CsANS1) and anthocyanidin reductase (CsANR1). Four types of microRNA/target modules, miR319b/CsTCP3-4, miR164b/CsCUC, miR396/CsGRF-GIF, and miR165b/HD-ZIPIII ones, were also identified and characterized for their functions in the regulation of the development of tea plant shoot tips and leaf shape. The results of these modules were reflected by their different expression patterns in developing buds and leaves that had distinctly different morphologies in three different tea plant varieties. Their roles in the regulation of catechin biosynthesis were also further verified by manipulation of microRNA319b (miR319b), which targets the transcripts of CsTCP3 and CsTCP4. Thus, CsTCPs represent at least one of these important groups of TFs that can integrate tea plant leaf development together with secondary metabolite biosynthesis. Our study provides new insight into shoot tip development and catechin production in tea plants and lays a foundation for further mechanistic understanding of the regulation of tea plant leaf development and secondary metabolism.

The size of leaf and seed organs, determined by the interplay of cell proliferation and expansion, is closely related to the final yield and quality of forage and crops. Yet the cellular and molecular mechanisms underlying organ size modulation remain poorly understood, especially in legumes. Here, MINI ORGAN1 (MIO1), which encodes an F-box protein SMALL LEAF AND BUSHY1 (SLB1) recently reported to control lateral branching in Medicago truncatula, was identified as a key regulator of organ size. We show that loss-of-function of MIO1/SLB1 severely reduced organ size. Conversely, plants overexpressing MIO1/SLB1 had enlarged organs. Cellular analysis revealed that MIO1/SLB1 controlled organ size mainly by modulating primary cell proliferation during the early stages of leaf development. Biochemical analysis revealed that MIO1/SLB1 could form part of SKP1/Cullin/F-box (SCF) E3 ubiquitin ligase complex, to target BIG SEEDS1 (BS1), a repressor of primary cell division, for degradation. Interestingly, we found that MIO1/SLB1 also played a key role in pulvinus development and leaf movement by modulating cell proliferation of the pulvinus as leaves developed. Our study not only demonstrates a conserved role of MIO1/SLB1 in the control of organ size in legumes, but also sheds light on the novel function of MIO1/SLB1 in leaf movement., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

In most legumes, two typical features found in leaves are diverse compound forms and the pulvinus-driven nyctinastic movement. Many genes have been identified for leaf-shape determination, but the underlying nature of leaf movement as well as its association with the compound form remains largely unknown. Using forward-genetic screening and whole-genome resequencing, we found that two allelic mutants of Medicago truncatula with unclosed leaflets at night were impaired in MtDWARF4A (MtDWF4A), a gene encoding a cytochrome P450 protein orthologous to Arabidopsis DWARF4. The mtdwf4a mutant also had a mild brassinosteroid (BR)-deficient phenotype bearing pulvini without significant deficiency in organ identity. Both mtdwf4a and dwf4 could be fully rescued by MtDWF4A, and mtdwf4a could close their leaflets at night after the application of exogenous 24-epi-BL. Surgical experiments and genetic analysis of double mutants revealed that the failure to exhibit leaf movement in mtdwf4a is a consequence of the physical obstruction of the overlapping leaflet laminae, suggesting a proper geometry of leaflets is important for their movement in M. truncatula. These observations provide a novel insight into the nyctinastic movement of compound leaves, shedding light on the importance of open space for organ movements in plants., (© 2021 The Authors. New Phytologist © 2021 New Phytologist Foundation.)

Plant-specific WOX family transcription factors play important roles ranging from embryogenesis to lateral organ development. The WOX1 transcription factors, which belong to the modern clade of the WOX family, are known to regulate outgrowth of the leaf blade specifically in the mediolateral axis; however, the role of WOX1 in compound leaf development remains unknown. Phylogenetic analysis of the whole WOX family in tomato (Solanum lycopersicum) indicates that there are 10 members that represent the modern, intermediate, and ancient clades. Using phylogenetic analysis and a reverse genetic approach, in this study we identified SlLAM1 in the modern clade and examined its function and tissue-specific expression pattern. We found that knocking out SlLAM1 via CRISPR/Cas9-mediated genome editing led to narrow leaves and a reduced number of secondary leaflets. Overexpression of tomato SlLAM1 could rescue the defects of the tobacco lam1 mutant. Anatomical and transcriptomic analyses demonstrated that floral organ development, fruit size, secondary leaflet initiation, and leaf complexity were altered due to loss-of-function of SlLAM1. These findings demonstrate that tomato SlLAM1 plays an important role in the regulation of secondary leaflet initiation, in addition to its conserved function in blade expansion., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

Food handlers are important sources of intestinal parasitic infection to the public and mass catering service areas. Several reports worldwide particularly in developing countries showed a high prevalence of intestinal parasitic infections among these groups of individuals. In an attempt to determine the prevalence of intestinal parasites, a cross-sectional study was carried out among food handlers in food establishment areas of Mettu town, Southwest Ethiopia. To collect sociodemographic characteristics of the study participants, a structured questionnaire and physical observation were employed. Stool specimens collected from each food handlers participated in the study were then examined using light microscopy of the wet mount followed by formol-ether concentration methods to see infection status. A total of 139 food handlers were enrolled in the study. Majority of them were females 89 (64%), and 124 (89.2%) of them had not taken any training related to food handling and preparation. Sixty-two (44.6%) of the study participants were diagnosed with one or more intestinal parasites. The chi-square test showed that intestinal parasite infection was found associated with personal hygiene ( p  ≤ 0.0001), nail trimming status ( p  ≤ 0.0001), and hair cover status ( p  ≤ 0.040). The multivariable logistic regression indicated that the risk to be infected with intestinal parasites were related with older age ( p  ≤ 0.032), food handlers who had no hand wash practices ( p  ≤ 0.033), who had no food handling and preparation training ( p  ≤ 0.005), poor personal hygiene ( p  ≤ 0.0001), who had not taken regular medical checkup ( p  ≤ 0.008), and whose nail was not trimmed ( p  ≤ 0.0001). The most abundant intestinal parasite identified was G. lamblia (24, 26.7%) followed by E. histolytica/dispar (22, 24.4%), and A. lumbricoides (15, 16.7%). Twenty-eight (45.2%) study individuals were also found with mixed infections. The study thus revealed a high prevalence of intestinal parasitic infection among apparently healthy food handlers in food establishments of the study area. Mass drug administration for all food handlers working in food establishment areas of the town coupled with health education and training should be initiated urgently., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2021 Solomon Yeshanew et al.)

WOX family transcription factors regulate multiple developmental programs. The intermediate clade transcriptional activator WOX9 functions together with the modern clade transcriptional repressor WOX genes in embryogenesis and meristems maintenance, but the mechanism of this interaction is unclear. STF and LAM1 are WOX1 orthologs required for leaf blade outgrowth in Medicago truncatula and Nicotiana sylvestris, respectively. Using biochemical methods and genome editing technology, here we show that WOX9 is an abaxial factor and functions antagonistically to STF and LAM1 to regulate leaf blade development. While NsWOX9 ectopic expression enhances the lam1 mutant phenotype, and antisense expression partially rescues the lam1 mutant, both overexpression and knockout of NsWOX9 in N. sylvestris resulted in a range of severe leaf blade distortions, indicating important role in blade development. Our results indicate that direct repression of WOX9 by WUS clade repressor STF/LAM1 is required for correct blade architecture and patterning in M. truncatula and N. sylvestris. These findings suggest that controlling transcriptional activation and repression mechanisms by direct interaction of activator and repressor WOX genes may be required for cell proliferation and differentiation homeostasis, and could be an evolutionarily conserved mechanism for the development of complex and diverse morphology in flowering plants., (© 2020 The Authors New Phytologist © 2020 New Phytologist Foundation.)

Key Message: A 3-ketoacyl-CoA synthase involved in biosynthesis of very long chain fatty acids and cuticular wax plays a vital role in aerial organ development in M. truncatula. Cuticular wax is composed of very long chain fatty acids and their derivatives. Defects in cuticular wax often result in organ fusion, but little is known about the role of cuticular wax in compound leaf and flower development in Medicago truncatula. In this study, through an extensive screen of a Tnt1 retrotransposon insertion population in M. truncatula, we identified four mutant lines, named wrinkled flower and leaf (wfl) for their phenotype. The phenotype of the wfl mutants is caused by a Tnt1 insertion in Medtr3g105550, encoding 3-ketoacyl-CoA synthase (KCS), which functions as a rate-limiting enzyme in very long chain fatty acid elongation. Reverse transcription-quantitative PCR showed that WFL was broadly expressed in aerial organs of the wild type, such as leaves, floral organs, and the shoot apical meristem, but was expressed at lower levels in roots. In situ hybridization showed a similar expression pattern, mainly detecting the WFL transcript in epidermal cells of the shoot apical meristem, leaf primordia, and floral organs. The wfl mutant leaves showed sparser epicuticular wax crystals on the surface and increased water permeability compared with wild type. Further analysis showed that in wfl leaves, the percentage of C20:0, C22:0, and C24:0 fatty acids was significantly increased, the amount of cuticular wax was markedly reduced, and wax constituents were altered compared to the wild type. The reduced formation of cuticular wax and wax composition changes on the leaf surface might lead to the developmental defects observed in the wfl mutants. These findings suggest that WFL plays a key role in cuticular wax formation and in the late stage of leaf and flower development in M. truncatula.

Key Message: PvArf regulate proline biosynthesis by physically interacting with PvP5CS1 to improve salt tolerance in switchgrass. The genetic factors that contribute to stress resiliency are yet to be determined. Here, we identified three ADP-ribosylation factors, PvArf1, PvArf-B1C, and PvArf-related, which contribute to salinity tolerance in transgenic switchgrass (Panicum virgatum L.). Switchgrass overexpressing each of these genes produced approximately twofold more biomass than wild type (WT) under normal growth conditions. Transgenic plants accumulated modestly higher levels of proline under normal conditions, but this level was significantly increased under salt stress providing better protection to transgenic plants compared to WT. We found that PvArf genes induce proline biosynthesis genes under salt stress to positively regulate proline accumulation, and further demonstrated that PvArf physically interact with PvP5CS1. Moreover, the transcript levels of two key ROS-scavenging enzyme genes were significantly increased in the transgenic plants compared to WT, leading to reduced H 2 O 2 accumulation under salt stress conditions. PvArf genes also protect cells against stress-induced changes in Na + and K + ion concentrations. Our findings uncover that ADP-ribosylation factors are key determinants of biomass yield in switchgrass, and play pivotal roles in salinity tolerance by regulating genes involved in proline biosynthesis.

Male sterility is an important tool for plant breeding and hybrid seed production. Male-sterile mutants are largely due to an abnormal development of either the sporophytic or gametophytic anther tissues. Tapetum, a key sporophytic tissue, provides nutrients for pollen development, and its delayed degeneration induces pollen abortion. Numerous bHLH proteins have been documented to participate in the degeneration of the tapetum in angiosperms, but relatively little attention has been given to the evolution of the involved developmental pathways across the phylogeny of land plants. A combination of cellular, molecular, biochemical and evolutionary analyses was used to investigate the male fertility control in Medicago truncatula. We characterized the male-sterile mutant empty anther1 (ean1) and identified EAN1 as a tapetum-specific bHLH transcription factor necessary for tapetum degeneration. Our study uncovered an evolutionarily conserved recruitment of bHLH subfamily II and III(a + c)1 in the regulation of tapetum degeneration. EAN1 belongs to the subfamily II and specifically forms heterodimers with the subfamily III(a + c)1 members, which suggests a heterodimerization mechanism conserved in angiosperms. Our work suggested that the pathway of two tapetal-bHLH subfamilies is conserved in all land plants, and likely was established before the divergence of the spore-producing land plants., (© 2020 The Authors. New Phytologist © 2020 New Phytologist Trust.)

Background: After prematurity, intrauterine growth restriction (IUGR) is the second leading cause of perinatal mortality. IUGR has significant consequences in fetal, neonatal, and adult life. Currently, Ethiopia lacks information on IUGR's prevalence and its determinants. This study aimed to assess the proportion of IUGR at birth and its associated factors., Methods: A cross-sectional study was carried out among women who give birth in four hospitals of south Gonder zone from November 2018 to February 2019. Multi-stage sampling was applied to select the required samples. IUGR was assessed using a standardized cutoff percentile/mean for each measurement. Data were collected by trained MSc clinical midwives. Bi-variable and multivariable logistic analyses were deployed to identify the association., Results: A total of 803 maternity women were participating in this study with a response rate of 95%. The proportion of IUGR 23.5% (95% CI: 20.7-26.6), low birth weight 13.3%, small-for- gestational-age 19.7%,and preterm birth 23.16%. Women who was unable to read and write, (AOR; 2.46, 95% CI: 1.02-5.92), total family size ≥7 (AOR; 1.67, 95% CI: 1.04-2.66), maternal mid-upper arm circumference (MUAC) < 23 cm (AOR; 2.10, 95% CI: 1.39-3.01), body mass index (BMI) < 18.5 kg/m 2 (AOR; 2.57, 95% CI: 1.72-3.83), altitude > 3000 m (AOR; 1.89 95% CI: 1.19-3.01), small placental size (< 350 g) (AOR; 2.42, 95% CI: 1.67-3.54) and small-for-gestational-age (AOR; 1.94, 95% CI:1.86-4.52) were the most predictors of IUGR., Conclusions: IUGR was a major public health concern in this study. Women who were unable to read and write, small-for-gestational-age, maternal BMI < 18.5 kg/m 2 , family size ≥7, maternal MUAC < 23 cm, small placental size, and altitude > 3000 m were found the most predictor variables. Strengthen female education, nutritional intervention before and during pregnancy, and routine maternity care is critical. Further clinical follow-up research is essential which includes maternal, fetal, and placental gens., Competing Interests: Competing interestsWe declare that there is no competing interest with anyone else., (© The Author(s) 2020.)

DELAY OF GERMINATION1 (DOG1) is a primary regulator of seed dormancy. Accumulation of DOG1 in seeds leads to deep dormancy and delayed germination in Arabidopsis. B3 domain-containing transcriptional repressors HSI2/VAL1 and HSL1/VAL2 silence seed dormancy and enable the subsequent germination and seedling growth. However, the roles of HSI2 and HSL1 in regulation of DOG1 expression and seed dormancy remain elusive. Seed dormancy was analysed by measurement of maximum germination percentage of freshly harvested Arabidopsis seeds. In vivo protein-protein interaction analysis, ChIP-qPCR and EMSA were performed and suggested that HSI2 and HSL1 can form dimers to directly regulate DOG1. HSI2 and HSL1 dimers interact with RY elements at DOG1 promoter. Both B3 and PHD-like domains are required for enrichment of HSI2 and HSL1 at the DOG1 promoter. HSI2 and HSL1 recruit components of polycomb-group proteins, including CURLY LEAF (CLF) and LIKE HETERCHROMATIN PROTEIN 1 (LHP1), for consequent deposition of H3K27me3 marks, leading to repression of DOG1 expression. Our findings suggest that HSI2- and HSL1-dependent histone methylation plays critical roles in regulation of seed dormancy during seed germination and early seedling growth., (© 2020 The Authors. New Phytologist © 2020 New Phytologist Trust.)

In species with compound leaves, the positions of leaflet primordium initiation are associated with local peaks of auxin accumulation. However, the role of auxin during the late developmental stages and outgrowth of compound leaves remains largely unknown. Using genome resequencing approaches, we identified insertion sites at four alleles of the LATERAL LEAFLET SUPPRESSION1 (LLS1) gene, encoding the auxin biosynthetic enzyme YUCCA1 in Medicago truncatula. Linkage analysis and complementation tests showed that the lls1 mutant phenotypes were caused by the Tnt1 insertions that disrupted the LLS1 gene. The transcripts of LLS1 can be detected in primordia at early stages of leaf initiation and later in the basal regions of leaflets, and finally in vein tissues at late leaf developmental stages. Vein numbers and auxin content are reduced in the lls1-1 mutant. Analysis of the lls1 sgl1 and lls1 palm1 double mutants revealed that SGL1 is epistatic to LLS1, and LLS1 works with PALM1 in an independent pathway to regulate the growth of lateral leaflets. Our work demonstrates that the YUCCA1/YUCCA4 subgroup plays very important roles in the outgrowth of lateral leaflets during compound leaf development of M. truncatula, in addition to leaf venation., (© 2020 The Authors. New Phytologist © 2020 New Phytologist Trust.)

Organ size is a major agronomic trait that determines grain yield and biomass production in crops. However, the molecular mechanisms controlling organ size, especially in legumes, are poorly understood. Using forward genetic approaches in a Tnt1 insertion mutant population of the model legume Medicago truncatula, we identified SMALL LEAF AND BUSHY1 (SLB1), which is required for the control of organ size and lateral branching. Loss of function of SLB1 led to reduced leaf and flower size but increased lateral branch formation in M. truncatula. SLB1 encodes an F-box protein, an orthologue of Arabidopsis thaliana STERILE APETALA (SAP), that forms part of an SKP1/Cullin/F-box E3 ubiquitin ligase complex. Biochemical and genetic analyses revealed that SLB1 controls M. truncatula organ growth and lateral branching by modulating the stability of BIG SEEDS1 (BS1). Moreover, the overexpression of SLB1 increased seed and leaf size in both M. truncatula and soybean (Glycine max), indicating functional conservation. Our findings revealed a novel mechanism by which SLB1 targets BS1 for degradation to regulate M. truncatula organ size and shoot branching, providing a new genetic tool for increasing seed yield and biomass production in crop and forage legumes., (© 2020 The Authors New Phytologist © 2020 New Phytologist Trust.)

Background: In the globe, 1.3 million deaths and around 50 million non-fatal injuries were reported. From all deaths, 90% occur in developing countries. Ethiopia is considered as one of the worst countries in the globe where road traffic accident causes a lot of fatalities and injuries of road users every year. The main objective of the study was to identify the main predictors of fatal car accident., Methods: The retrospective research design was applied. 255 records were taken from Finote Selam traffic police office, northwest part of Ethiopia from September 2009 to January 2018. The statistical analysis was performed by using SPSS version 23 software. Chi-square for association test and ordinal logistic regression for predictor identification were used., Results: Age of drivers were the responsible causes of fatal road traffic accident (p-value = 0.033). The more experienced drivers decreased the occurrence of fatal traffic accidents. In addition, increasing vehicle service year reduced the occurrence of accidental death. Besides, the occurrence of fatal car accident in autumn season was 0.44 times less than that of in summer season. Additionally, drivers' educational level was played a crucial role in a road traffic accident. For instance, drivers whose educational level was below 12th grade were the most responsible factor for car accident deaths. What is more, it was seen that drivers who drove their vehicles could minimize the occurrence of fatal traffic accident (p-value = 0.010)., Conclusion: In conclusion, fatal road traffic accidents happened due to drivers' lack of sufficient driving experience and low educational level. In addition, driving on weekends and driving on summer season were disclosed as responsible for fatal car accident. Moreover, drivers with younger age and those who drove a new vehicle likely caused fatal car accident. However, drivers who drive their vehicles seemed to be less responsible for fatal car accident than that of employed.

Compound leaves show more complex patterns than simple leaves, and this is mainly because of a specific morphogenetic process (leaflet initiation and arrangement) that occurs during their development. How the relevant morphogenetic activity is established and modulated to form a proper pattern of leaflets is a central question. Here we show that the trifoliate leaf pattern of the model leguminous plant Medicago truncatula is controlled by the BEL1-like homeodomain protein PINNATE-LIKE PENTAFOLIATA1 (PINNA1). We identify PINNA1 as a determinacy factor during leaf morphogenesis that directly represses transcription of the LEAFY (LFY) orthologue SINGLE LEAFLET1 (SGL1), which encodes an indeterminacy factor key to the morphogenetic activity maintenance. PINNA1 functions alone in the terminal leaflet region and synergizes with another determinacy factor, the C2H2 zinc finger protein PALMATE-LIKE PENTAFOLIATA1 (PALM1), in the lateral leaflet regions to define the spatiotemporal expression of SGL1, leading to an elaborate control of morphogenetic activity. This study reveals a framework for trifoliate leaf-pattern formation and sheds light on mechanisms generating diverse leaf forms.

The formation of nitrogen-fixing nodules on legume hosts is a finely tuned process involving many components of both symbiotic partners. Production of the exopolysaccharide succinoglycan by the nitrogen-fixing bacterium Sinorhizobium meliloti 1021 is needed for an effective symbiosis with Medicago spp., and the succinyl modification to this polysaccharide is critical. However, it is not known when succinoglycan intervenes in the symbiotic process, and it is not known whether the plant lysin-motif receptor-like kinase MtLYK10 intervenes in recognition of succinoglycan, as might be inferred from work on the Lotus japonicus MtLYK10 ortholog, LjEPR3. We studied the symbiotic infection phenotypes of S. meliloti mutants deficient in succinoglycan production or producing modified succinoglycan, in wild-type Medicago truncatula plants and in Mtlyk10 mutant plants. On wild-type plants, S. meliloti strains producing no succinoglycan or only unsuccinylated succinoglycan still induced nodule primordia and epidermal infections, but further progression of the symbiotic process was blocked. These S. meliloti mutants induced a more severe infection phenotype on Mtlyk10 mutant plants. Nodulation by succinoglycan-defective strains was achieved by in trans rescue with a Nod factor-deficient S. meliloti mutant. While the Nod factor-deficient strain was always more abundant inside nodules, the succinoglycan-deficient strain was more efficient than the strain producing only unsuccinylated succinoglycan. Together, these data show that succinylated succinoglycan is essential for infection thread formation in M. truncatula, and that MtLYK10 plays an important, but different role in this symbiotic process. These data also suggest that succinoglycan is more important than Nod factors for bacterial survival inside nodules., (© 2019 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Emerging evidence shows that the recent pandemic of coronavirus disease 19 (COVID-19) is characterized by coagulation activation and endothelial dysfunction. This increases the risk of morbidity, mortality and economic loss among COVID-19 patients. Therefore, there was an urgent need to investigate the extent and risk factors of thromboembolism among COVID-19 patients. English-language based databases (PubMed, Cumulative Index to Nursing and Allied Health Literature, EMBASE, and Cochrane library) were exhaustively searched to identify studies related to prevalence of thromboembolism among hospitalized COVID-19 patients. A random-effects model was employed to estimate the pooled prevalence of thromboembolism. The pooled prevalence of thrombotic events was computed using STATA 16.0 software. Heterogeneity analysis was reported using I 2 . A total of 19 studies with 2,520 patients with COVID-19 were included. The pooled prevalence of thrombotic events of hospitalized patients with COVID-19 was 33% (95% CI: 25-41%, I 2 = 97.30%, p < 0.001) with a high degree of heterogeneity across studies. Elevated D-dimer hospitalized in the intensive care unit and being under mechanical ventilation were the most frequently associated factors for the development of thrombotic events. The pooled prevalence of thrombotic events in COVID-19 patients was 33%. The prevalence of thrombotic event is variables on the basis of study design and study centers. Several risk factors such as, elevated D-dimer, hospitalized in the intensive care unit and being under mechanical ventilation, were the most frequently reported risk factors identified. Therefore, healthcare professionals should consider these risk factors to optimally manage thromboembolism in COVID-19 patients.

Carotenoids are a group of natural tetraterpenoid pigments with indispensable roles in the plant life cycle and the human diet. Although the carotenoid biosynthetic pathway has been well characterized, the regulatory mechanisms that control carotenoid metabolism, especially in floral organs, remain poorly understood. In this study, we identified an anthocyanin-related R2R3-MYB protein, WHITE PETAL1 (WP1), that plays a critical role in regulating floral carotenoid pigmentation in Medicago truncatula Carotenoid analyses showed that the yellow petals of the wild-type M. truncatula contained high concentrations of carotenoids that largely consisted of esterified lutein and that disruption of WP1 function via Tnt1 insertion led to substantially reduced lutein accumulation. WP1 mainly functions as a transcriptional activator and directly regulates the expression of carotenoid biosynthetic genes including MtLYCe and MtLYCb through its C-terminal acidic activation motif. Further molecular and genetic analyses revealed that WP1 physically interacts with MtTT8 and MtWD40-1 proteins and that this interaction facilitates WP1's function in the transcriptional activation of both carotenoid and anthocyanin biosynthetic genes. Our findings demonstrate the molecular mechanism of WP1-mediated regulation of floral carotenoid pigmentation and suggest that the conserved MYB-basic-helix-loop-helix-WD40 regulatory module functions in carotenoid biosynthesis in M. truncatula , with specificity imposed by the MYB partner., (© 2019 American Society of Plant Biologists. All rights reserved.)

Plant lateral organ development is a complex process involving both transcriptional activation and repression mechanisms. The WOX transcriptional repressor WOX1/STF, the LEUNIG (LUG) transcriptional corepressor and the ANGUSTIFOLIA3 (AN3) transcriptional coactivator play important roles in leaf blade outgrowth and flower development, but how these factors coordinate their activities remains unclear. Here we report physical and genetic interactions among these key regulators of leaf and flower development. We developed a novel in planta transcriptional activation/repression assay and suggest that LUG could function as a transcriptional coactivator during leaf blade development. MtLUG physically interacts with MtAN3, and this interaction appears to be required for leaf and flower development. A single amino acid substitution at position 61 in the SNH domain of MtAN3 protein abolishes its interaction with MtLUG, and its transactivation activity and biological function. Mutations in lug and an3 enhanced each other's mutant phenotypes. Both the lug and the an3 mutations enhanced the wox1 prs leaf and flower phenotypes in Arabidopsis. Our findings together suggest that transcriptional repression and activation mediated by the WOX, LUG and AN3 regulators function in concert to promote leaf and flower development, providing novel mechanistic insights into the complex regulation of plant lateral organ development., (© 2019 The Authors. New Phytologist © 2019 New Phytologist Trust.)

Objective: The aim of this study was to determine the major risk factors of antiretroviral therapy dropout. The retrospective cohort research design was applied. 1512 HIV patients were included from Mettu Karl Hospital in Illubabor Zone, southwest part of Ethiopia from September 2005 to January 2018. Kaplan-Meier comparison and log-logistic regression accelerated failure time model were used., Results: From the log-logistic regression result, the risk of dropout for patients with primary education status was 10.58% greater as compared to illiterate (p < 0.0110). The probability of dropout for patients with marital status separated was about 16.82% higher than those patients with marital status divorced (p < 0.0070). Being merchant, farmer and daily labour had a greater risk of dropout as compared to a housewife. Most of the HIV/AIDS patients on ART were dropout in a short period due to patients separated marital status, primary education, CD4, being merchants, farmer and daily labour. Investigation on the cause of antiretroviral therapy dropout from a number of AIDS clinics in the country is highly appreciated.

The formation and maintenance of the shoot apical meristem (SAM) are critical for plant development. However, the underlying molecular mechanism of regulating meristematic cell activity is poorly understood in the model legume Medicago truncatula. Using forward genetic approaches, we identified HEADLESS (HDL), a homolog of Arabidopsis WUSCHEL, required for SAM maintenance and leaf development in M. truncatula. Disruption of HDL led to disorganized specification and arrest of the SAM and axillary meristems, resulting in the hdl mutant being locked in the vegetative phase without apparent stem elongation. hdl mutant leaves are shorter in the proximal-distal axis due to reduced leaf length elongation, which resulted in a higher blade width/length ratio and altered leaf shape, uncovering novel phenotypes undescribed in the Arabidopsis wus mutant. HDL functions as a transcriptional repressor by recruiting MtTPL through its conserved WUS-box and EAR-like motif. Further genetic analysis revealed that HDL and STENOFOLIA (STF), a key regulator of M. truncatula lamina outgrowth, act independently in leaf development although HDL could recruit MtTPL in the same manner as STF does. Our results indicate that HDL has conserved and novel functions in regulating shoot meristems and leaf shape in M. truncatula, providing new avenues for understanding meristem biology and plant development.

Objective: The purpose of this study was to identify the major risk factors, which contributed to shortened survival time to death of HIV patients on antiretroviral therapy. Six-hundred HIV patients were included from two hospitals and six health centers record from January 2003 to December 2017. Kaplan-Meier and Cox proportional hazard model were implemented., Results: From the Kaplan-Meier, log-rank test result indicated that there was a significant difference between tuberculosis comorbidity (P = .000), occupation (P = .027), and WHO clinical stage (P = .012) on the survival experience of patients at 5% statistical significance level. From the Cox regression result, the risk of death for patients who lived with tuberculosis was about 2.872-fold times higher than those patients who were negative. Most of the HIV/AIDS patients on antiretroviral therapy were died in a short period due to tuberculosis comorbidity, began with lower amount of CD4, being underweight, merchant, and being on WHO clinical stage IV.