Aromatic amino acid biosynthesis impacts root hair development and symbiotic associations in Lotus japonicus.

Legume roots can be symbiotically colonized by arbuscular mycorrhizal (AM) fungi and nitrogen-fixing bacteria. In Lotus japonicus, the latter occurs intracellularly by the cognate rhizobial partner Mesorhizobium loti or intercellularly with the Agrobacterium pusense strain IRBG74. Although these symbiotic programs show distinctive cellular and transcriptome signatures, some molecular components are shared. In this study, we demonstrate that 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase 1 (DAHPS1), the first enzyme in the biosynthetic pathway of aromatic amino acids (AAAs), plays a critical role in root hair development and for AM and rhizobial symbioses in Lotus. Two homozygous DAHPS1 mutants (dahps1-1 and dahps1-2) showed drastic alterations in root hair morphology, associated with alterations in cell wall dynamics and a progressive disruption of the actin cytoskeleton. The altered root hair structure was prevented by pharmacological and genetic complementation. dahps1-1 and dahps1-2 showed significant reductions in rhizobial infection (intracellular and intercellular) and nodule organogenesis and a delay in AM colonization. RNAseq analysis of dahps1-2 roots suggested that these phenotypes are associated with downregulation of several cell wall-related genes, and with an attenuated signaling response. Interestingly, the dahps1 mutants showed no detectable pleiotropic effects, suggesting a more selective recruitment of this gene in certain biological processes. This work provides robust evidence linking AAA metabolism to root hair development and successful symbiotic associations.

Cyber sexual harassment against women and impact on health. A cross-sectional study in a representative population sample.

BACKGROUND: This study focuses on the frequency of cyber sexual harassment (CSH) and examines its link with five health impact indicators (self-perceived health, medication, suicidal ideation, suicide attempts and use of health services). METHODS: Analyzing the data provided by the 2019 Macro-survey on violence against women in Spain, two items of which refer to CSH. It was conducted in Spain on a representative sample of 9568 women aged over 16 years old. RESULTS: A total of 9.15% of the women surveyed had experienced CSH at some point in their lives. Being under 25 years old, having higher education, not being in a relationship, having no religious beliefs and having a certified disability are sociodemographic characteristics associated with a higher risk of CSH. Women who have experienced other forms of gender-based violence also show an increased risk. Female victims of CSH reported higher rates of suicidal ideation (20% versus 9.79% in non-victims of CSH) and suicidal attempts (7.20% versus 1.74% in non-victims of CSH). CONCLUSIONS: These findings have significant implications for the design of preventive health polices, which should incorporate strategies to address CSH as part of the continuum of multiple interrelated forms of gender violence that affect women and girls throughout their lives.

Systematic review of healthcare interventions for reducing gender-based violence impact on the mental health of women with disabilities.

PURPOSE: Women with disabilities are more exposed to violence. The health sector has a key role in all three levels of prevention of violence against women. The objective of this paper was to review the interventions for preventing gender-based violence and reducing its impact on the mental health of women with any form of disability. METHOD: Relevant studies were identified through conducting searches in PubMed, Scopus, CINAHL, PsyInfo, Social Services Abstracts, and PILOTS. Two reviewers analyzed and selected studies. A qualitative synthesis was made. RESULTS: 3149 references were obtained, among which eight articles describing nine interventions from the USA and the UK. Most were intended for women with mental/intellectual disability and assessed intimate partner or sexual violence. Only one study showed high methodological quality. They were found to be particularly effective as regards improvement of the skills acquired by participants, but the results as regards improved mental health are not consistent. CONCLUSION: Our review shows very little evidence of effective interventions. Further studies are required with higher internal validity and female sample groups with diverse disabilities. CLINICAL RELEVANCE: Gender-based violence is a highly prevalent problem for women with disabilities, and in addition to being a public health challenge is a violation of human rights. Health care systems and policymakers should take a key role in all three levels of prevention of violence against women with disabilities. Interventions with longer follow-up times are required. It is also important for interventions to be designed in consultation with people with disabilities.

A Promiscuity Locus Confers Lotus burttii Nodulation with Rhizobia from Five Different Genera.

Legumes acquire access to atmospheric nitrogen through nitrogen fixation by rhizobia in root nodules. Rhizobia are soil-dwelling bacteria and there is a tremendous diversity of rhizobial species in different habitats. From the legume perspective, host range is a compromise between the ability to colonize new habitats, in which the preferred symbiotic partner may be absent, and guarding against infection by suboptimal nitrogen fixers. Here, we investigate natural variation in rhizobial host range across Lotus species. We find that Lotus burttii is considerably more promiscuous than Lotus japonicus, represented by the Gifu accession, in its interactions with rhizobia. This promiscuity allows Lotus burttii to form nodules with Mesorhizobium, Rhizobium, Sinorhizobium, Bradyrhizobium, and Allorhizobium species that represent five distinct genera. Using recombinant inbred lines, we have mapped the Gifu/burttii promiscuity quantitative trait loci (QTL) to the same genetic locus regardless of rhizobial genus, suggesting a general genetic mechanism for symbiont-range expansion. The Gifu/burttii QTL now provides an opportunity for genetic and mechanistic understanding of promiscuous legume-rhizobia interactions. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Distinct signaling routes mediate intercellular and intracellular rhizobial infection in Lotus japonicus.

Rhizobial infection of legume roots during the development of nitrogen-fixing root nodules can occur intracellularly, through plant-derived infection threads traversing cells, or intercellularly, via bacterial entry between epidermal plant cells. Although it is estimated that around 25% of all legume genera are intercellularly infected, the pathways and mechanisms supporting this process have remained virtually unexplored due to a lack of genetically amenable legumes that exhibit this form of infection. In this study, we report that the model legume Lotus japonicus is infected intercellularly by the IRBG74 strain, recently proposed to belong to the Agrobacterium clade of the Rhizobiaceae. We demonstrate that the resources available for L. japonicus enable insight into the genetic requirements and fine-tuning of the pathway governing intercellular infection in this species. Inoculation of L. japonicus mutants shows that Ethylene-responsive factor required for nodulation 1 (Ern1) and Leu-rich Repeat Receptor-Like Kinase (RinRK1) are dispensable for intercellular infection in contrast to intracellular infection. Other symbiotic genes, including nod factor receptor 5 (NFR5), symbiosis receptor-like kinase (SymRK), Ca2+/calmodulin dependent kinase (CCaMK), exopolysaccharide receptor 3 (Epr3), Cyclops, nodule inception (Nin), nodulation signaling pathway 1 (Nsp1), nodulation signaling pathway 2 (Nsp2), cystathionine-ß-synthase (Cbs), and Vapyrin are equally important for both entry modes. Comparative RNAseq analysis of roots inoculated with IRBG74 revealed a distinctive transcriptome response compared with intracellular colonization. In particular, several cytokinin-related genes were differentially regulated. Corroborating this observation, cyp735A and ipt4 cytokinin biosynthesis mutants were significantly affected in their nodulation with IRBG74, whereas lhk1 cytokinin receptor mutants formed no nodules. These results indicate a differential requirement for cytokinin signaling during intercellular rhizobial entry and highlight distinct modalities of inter- and intracellular infection mechanisms in L. japonicus.

Intimate partner violence during pregnancy and risk of fetal and neonatal death: a meta-analysis with socioeconomic context indicators.

OBJECTIVE: The objective of the study was to summarize the results from observational studies examining the risk of fetal and neonatal death (perinatal death) as a function of the experience of intimate partner violence during pregnancy and examine the influence of socioeconomic context indicators on this association. DATA SOURCES: Bibliographic searches were conducted in PubMed, EMBASE, CINAHL, and LILACS until March 2019. STUDY ELIGIBILITY CRITERIA: We considered observational studies that provided data on the association between intimate partner violence during pregnancy and perinatal death. STUDY APPRAISAL AND SYNTHESIS METHODS: Information collected included study characteristics, type, and prevalence of intimate partner violence and the reported association between intimate partner violence and perinatal death. Quality of the included studies was assessed using the Newcastle-Ottawa scale. Two reviewers independently conducted all review procedures; disagreements were resolved by a third reviewer. Meta-analyses were conducted based on the specific type of intimate partner violence (physical, psychological, sexual, unspecified) and also based on any type of intimate partner violence, considering 1 effect size per study, regardless of the type of intimate partner violence analyzed. Meta-regression analyses were performed to assess the possible effects of socioeconomic context. The proportion of deaths attributable to the exposure of intimate partner violence based on the crude data from the 3 cohort studies available also was calculated. RESULTS: Seventeen studies were included. The random-effects model showed a statistically significant increase in the odds of perinatal death among women exposed to unspecified intimate partner violence (odds ratio, 3.18; 95% confidence interval, 1.88-5.38), physical intimate partner violence (odds ratio, 2.46; 95% confidence interval, 1.76-3.44), and any type of intimate partner violence during pregnancy (odds ratio, 2.89; 95% confidence interval, 2.03-4.10). Meta-regression analysis showed stronger associations in countries with higher gross domestic product (odds ratio, 1.03; 95% confidence interval, 1.02-1.04) and a higher percentage of health expenditure (odds ratio, 1.27; 95% confidence interval, 1.09-1.46). The proportion of deaths attributable to exposure to intimate partner violence in cohort studies was attributable proportion, 60%; 95% confidence interval, 15-81%. CONCLUSION: Pregnant women who experience intimate partner violence during pregnancy may be about 3 times more likely to suffer perinatal death compared with women who do not experience intimate partner violence. It should be a priority to include intimate partner violence screenings or other detection strategies in pregnancy monitoring or family-planning programs because these could help avoid preventable perinatal deaths.

Morphotype of bacteroids in different legumes correlates with the number and type of symbiotic NCR peptides.

In legume nodules, rhizobia differentiate into nitrogen-fixing forms called bacteroids, which are enclosed by a plant membrane in an organelle-like structure called the symbiosome. In the Inverted Repeat-Lacking Clade (IRLC) of legumes, this differentiation is terminal due to irreversible loss of cell division ability and is associated with genome amplification and different morphologies of the bacteroids that can be swollen, elongated, spherical, and elongated-branched, depending on the host plant. In Medicago truncatula, this process is orchestrated by nodule-specific cysteine-rich peptides (NCRs) delivered into developing bacteroids. Here, we identified the predicted NCR proteins in 10 legumes representing different subclades of the IRLC with distinct bacteroid morphotypes. Analysis of their expression and predicted sequences establishes correlations between the composition of the NCR family and the morphotypes of bacteroids. Although NCRs have a single origin, their evolution has followed different routes in individual lineages, and enrichment and diversification of cationic peptides has resulted in the ability to impose major morphological changes on the endosymbionts. The wide range of effects provoked by NCRs such as cell enlargement, membrane alterations and permeabilization, and biofilm and vesicle formation is dependent on the amino acid composition and charge of the peptides. These effects are strongly influenced by the rhizobial surface polysaccharides that affect NCR-induced differentiation and survival of rhizobia in nodule cells.

Transcriptome analysis of the differential effect of the NADPH oxidase gene RbohB in Phaseolus vulgaris roots following Rhizobium tropici and Rhizophagus irregularis inoculation.

BACKGROUND: Reactive oxygen species (ROS) are generated by NADPH oxidases known as respiratory burst oxidase homologs (RBOHs) in plants. ROS regulate various cellular processes, including the mutualistic interactions between legumes and nitrogen-fixing bacteria or arbuscular mycorrhizal (AM) fungi. Rboh is a multigene family comprising nine members (RbohA-I) in common bean (Phaseolus vulgaris). The RNA interference-mediated silencing of RbohB (PvRbohB-RNAi) in this species diminished its ROS production and greatly impaired nodulation. By contrast, the PvRbohB-RNAi transgenic roots showed early hyphal root colonization with enlarged fungal hypopodia; therefore, we proposed that PvRbohB positively regulates rhizobial infection (Rhizobium tropici) and inhibits AM colonization by Rhizophagus irregularis in P. vulgaris. RESULTS: To corroborate this hypothesis, an RNA-Seq transcriptomic analysis was performed to identify the differentially expressed genes in the PvRbohB-RNAi roots inoculated with Rhizobium tropici or Rhizophagus irregularis. We found that, in the early stages, root nodule symbioses generated larger changes of the transcriptome than did AM symbioses in P. vulgaris. Genes related to ROS homeostasis and cell wall flexibility were markedly upregulated in the early stages of rhizobial colonization, but not during AM colonization. Compared with AM colonization, the rhizobia induced the expression of a greater number of genes encoding enzymes involved in the metabolism of auxins, cytokinins, and ethylene, which were typically repressed in the PvRbohB-RNAi roots. CONCLUSIONS: Our research provides substantial insights into the genetic interaction networks in the early stages of rhizobia and AM symbioses with P. vulgaris, as well as the differential roles that RbohB plays in processes related to ROS scavenging, cell wall remodeling, and phytohormone homeostasis during nodulation and mycorrhization in this legume.

Root stem cell niche maintenance and apical meristem activity critically depend on THREONINE SYNTHASE1.

Indeterminate root growth depends on the stem cell niche (SCN) and root apical meristem (RAM) maintenance whose regulation permits plasticity in root system formation. Using a forward genetics approach, we isolated the moots koom1 ('short root' in Mayan) mutant that shows complete primary RAM exhaustion and abolished SCN activity. We identified that this phenotype is caused by a point mutation in the METHIONINE OVERACCUMULATOR2 (MTO2) gene that encodes THREONINE SYNTHASE1 and renamed the mutant as mto2-2. The amino acid profile showed drastic changes, most notorious of which was accumulation of methionine. In non-allelic mto1-1 (Arabidopsis thaliana cystathionine gamma-synthetase1) and mto3-1 (S-adenosylmethionine synthetase) mutants, both with an increased methionine level, the RAM size was similar to that of the wild type, suggesting that methionine overaccumulation itself did not cause RAM exhaustion in mto2 mutants. When mto2-2 RAM is not yet completely exhausted, exogenous threonine induced de novo SCN establishment and root growth recovery. The threonine-dependent RAM re-establishment in mto2-2 suggests that threonine is a limiting factor for RAM maintenance. In the root, MTO2 was predominantly expressed in the RAM. The essential role of threonine in mouse embryonic stem cells and in RAM maintenance suggests that common regulatory mechanisms may operate in plant and animal SCN maintenance.

Terminal Bacteroid Differentiation Is Associated With Variable Morphological Changes in Legume Species Belonging to the Inverted Repeat-Lacking Clade.

Medicago and closely related legume species from the inverted repeat-lacking clade (IRLC) impose terminal differentiation onto their bacterial endosymbionts, manifested in genome endoreduplication, cell enlargement, and loss of cell-division capacity. Nodule-specific cysteine-rich (NCR) secreted host peptides are plant effectors of this process. As bacteroids in other IRLC legumes, such as Cicer arietinum and Glycyrrhiza lepidota, were reported not to display features of terminal differentiation, we investigated the fate of bacteroids in species from these genera as well as in four other species representing distinct genera of the phylogenetic tree for this clade. Bacteroids in all tested legumes proved to be larger in size and DNA content than cultured cells; however, the degree of cell elongation was rather variable in the different species. In addition, the reproductive ability of the bacteroids isolated from these legumes was remarkably reduced. In all IRLC species with available sequence data, the existence of NCR genes was found. These results indicate that IRLC legumes provoke terminal differentiation of their endosymbionts with different morphotypes, probably with the help of NCR peptides.

Legume NADPH Oxidases Have Crucial Roles at Different Stages of Nodulation.

Plant NADPH oxidases, formerly known as respiratory burst oxidase homologues (RBOHs), are plasma membrane enzymes dedicated to reactive oxygen species (ROS) production. These oxidases are implicated in a wide variety of processes, ranging from tissue and organ growth and development to signaling pathways in response to abiotic and biotic stimuli. Research on the roles of RBOHs in the plant's response to biotic stresses has mainly focused on plant-pathogen interactions; nonetheless, recent findings have shown that these oxidases are also involved in the legume-rhizobia symbiosis. The legume-rhizobia symbiosis leads to the formation of the root nodule, where rhizobia reduce atmospheric nitrogen to ammonia. A complex signaling and developmental pathway in the legume root hair and root facilitate rhizobial entrance and nodule organogenesis, respectively. Interestingly, several reports demonstrate that RBOH-mediated ROS production displays versatile roles at different stages of nodulation. The evidence collected to date indicates that ROS act as signaling molecules that regulate rhizobial invasion and also function in nodule senescence. This review summarizes discoveries that support the key and versatile roles of various RBOH members in the legume-rhizobia symbiosis.

PvRbohB negatively regulates Rhizophagus irregularis colonization in Phaseolus vulgaris.

Plant NADPH oxidases (RBOHs) regulate the early stages of rhizobial infection in Phaseolus vulgaris and affect nodule function in Medicago truncatula. In contrast, the role of RBOHs in the plant-arbuscular mycorrhizal (AM) symbiosis and in the regulation of reactive oxygen species (ROS) production during the establishment of the AM interaction is largely unknown. In this study, we assessed the role of P. vulgaris Rboh (PvRbohB) during the symbiosis with the AM fungus, Rhizophagus irregularis. Our results indicate that the PvRbohB transcript is significantly up-regulated in the mycorrhized roots of P. vulgaris. Further, the PvRbohB promoter was found to be active during the invasion of R. irregularis. Down-regulation of PvRbohB transcription by RNAi (RNA interference) silencing resulted in diminished ROS levels in the transgenic mycorrhized roots and induced early hyphal root colonization. Interestingly, the size of appressoria increased in PvRbohB-RNAi roots (760 ± 70.1 µm) relative to controls (251 ± 73.2 µm). Finally, the overall level of mycorrhizal colonization significantly increased in PvRbohB-RNAi roots [48.1 ± 3.3% root length colonization (RLC)] compared with controls (29.4 ± 1.9% RLC). We propose that PvRbohB negatively regulates AM colonization in P. vulgaris.

Determinate primary root growth as an adaptation to aridity in Cactaceae: towards an understanding of the evolution and genetic control of the trait.

BACKGROUND AND AIMS: Species of Cactaceae are well adapted to arid habitats. Determinate growth of the primary root, which involves early and complete root apical meristem (RAM) exhaustion and differentiation of cells at the root tip, has been reported for some Cactoideae species as a root adaptation to aridity. In this study, the primary root growth patterns of Cactaceae taxa from diverse habitats are classified as being determinate or indeterminate, and the molecular mechanisms underlying RAM maintenance in Cactaceae are explored. Genes that were induced in the primary root of Stenocereus gummosus before RAM exhaustion are identified. METHODS: Primary root growth was analysed in Cactaceae seedlings cultivated in vertically oriented Petri dishes. Differentially expressed transcripts were identified after reverse northern blots of clones from a suppression subtractive hybridization cDNA library. KEY RESULTS: All species analysed from six tribes of the Cactoideae subfamily that inhabit arid and semi-arid regions exhibited determinate primary root growth. However, species from the Hylocereeae tribe, which inhabit mesic regions, exhibited mostly indeterminate primary root growth. Preliminary results suggest that seedlings of members of the Opuntioideae subfamily have mostly determinate primary root growth, whereas those of the Maihuenioideae and Pereskioideae subfamilies have mostly indeterminate primary root growth. Seven selected transcripts encoding homologues of heat stress transcription factor B4, histone deacetylase, fibrillarin, phosphoethanolamine methyltransferase, cytochrome P450 and gibberellin-regulated protein were upregulated in S. gummosus root tips during the initial growth phase. CONCLUSIONS: Primary root growth in Cactoideae species matches their environment. The data imply that determinate growth of the primary root became fixed after separation of the Cactiodeae/Opuntioideae and Maihuenioideae/Pereskioideae lineages, and that the genetic regulation of RAM maintenance and its loss in Cactaceae is orchestrated by genes involved in the regulation of gene expression, signalling, and redox and hormonal responses.

A collection of novel Lotus japonicus LORE1 mutants perturbed in the nodulation program induced by the Agrobacterium pusense strain IRBG74.

The Lotus japonicus population carrying new Lotus retrotransposon 1 (LORE1) insertions represents a valuable biological resource for genetic research. New insertions were generated by activation of the endogenous retroelement LORE1a in the germline of the G329-3 plant line and arranged in a 2-D system for reverse genetics. LORE1 mutants identified in this collection contributes substantially to characterize candidate genes involved in symbiotic association of L. japonicus with its cognate symbiont, the nitrogen-fixing bacteria Mesorhizobium loti that infects root nodules intracellularly. In this study we aimed to identify novel players in the poorly explored intercellular infection induced by Agrobacterium pusense IRBG74 sp. For this purpose, a forward screen of > 200,000 LORE1 seedlings, obtained from bulk propagation of G329-3 plants, inoculated with IRBG74 was performed. Plants with perturbed nodulation were scored and the offspring were further tested on plates to confirm the symbiotic phenotype. A total of 110 Lotus mutants with impaired nodulation after inoculation with IRBG74 were obtained. A comparative analysis of nodulation kinetics in a subset of 20 mutants showed that most of the lines were predominantly affected in nodulation by IRBG74. Interestingly, additional defects in the main root growth were observed in some mutant lines. Sequencing of LORE1 flanking regions in 47 mutants revealed that 92 Lotus genes were disrupted by novel LORE1 insertions in these lines. In the IM-S34 mutant, one of the insertions was located in the 5´UTR of the LotjaGi5g1v0179800 gene, which encodes the AUTOPHAGY9 protein. Additional mutant alleles, named atg9-2 and atg9-3, were obtained in the reverse genetic collection. Nodule formation was significantly reduced in these mutant alleles after M. loti and IRBG74 inoculation, confirming the effectiveness of the mutant screening. This study describes an effective forward genetic approach to obtain novel mutants in Lotus with a phenotype of interest and to identify the causative gene(s).

Relationship between Sexual Violence and the Health of Spanish Women-A National Population-Based Study.

BACKGROUND: Health consequences are likely to be different when sexual violence is analysed independently from other types of violence. It is also likely that different health consequences will result in the cases of partner or ex-partner sexual violence, non-partner sexual violence and sexual harassment. METHODS: This study is based on the 2019 Macro-survey of Violence against Women conducted by the Spanish Ministry of Equality on a sample of 9568 women aged 16 years or older. Odds ratios were calculated, and multinomial logistic regression analyses were performed. RESULTS: The present study estimates that 4 out of 10 surveyed women had experienced some form of sexual violence in their lifetime. Sexual harassment is the most frequently reported form of this violence, while intimate partner sexual violence is the form with the most unfavourable sociodemographic characteristics and the worst health impact indicators, such as a greater likelihood of suicidal behaviour. CONCLUSIONS: Sexual violence is a widespread, under-studied problem with negative health impacts. Women exposed to intimate partner violence are the most vulnerable and at risk. It is advised that responses and comprehensive care plans be developed that place special emphasis on the protection of victims' mental health.

The prevalence of intimate partner violence against women with disabilities: a systematic review of the literature.

PURPOSE: Although systematic reviews have already been conducted on violence and disability, the evidence is not conclusive in Intimate Partner Violence (IPV). This study examined the prevalence of IPV in women with disabilities, taking account of the different types of violence and disability. METHODS: We performed a search in five databases. We included observational studies that analysed the frequency of IPV in women with disabilities compared to women without. Two independent reviewers selected and assessed studies. We made a qualitative synthesis according to the type of IPV analysed in relation to disability. RESULTS: We identified 26 articles. The frequency and risk of IPV were greater in women with disabilities than in those without. All of the articles that studied financial violence, 81.3% of those that studied physical violence, 78.5% of those that studied psychological violence, 75% of those that studied physical/sexual violence, 73.3% of those that studied sexual violence and 50% of those that studied any type of violence found a significant association whit disability. CONCLUSIONS: Women with disabilities are at higher risk of IPV. Rehabilitation centers and its professionals must have a leading role in the identification of these situations and this should be considered in IPV prevention plans.Implications for rehabilitationPeople with disabilities are at higher risk of multiple types of violence.Violence against women with disabilities therefore involves two public health issues interacting simultaneously.Rehabilitation centers and its professionals should have a leading role in the identification of IPV in women with disabilities.IPV prevention plans should consider rehabilitation centers and its professionals as a main component of interventions in women with disabilities.

Characteristics and effectiveness of interventions to reduce cyberbullying: a systematic review.

Background: This paper presents the results from a systematic review on the effectiveness of interventions to reduce Cyberbullying (CB) as a function of their outcomes and main characteristics; and an analysis of the level of completeness to which the characteristics of these interventions are described. Methods: Systematic searches were conducted in PubMed, Scopus, ERIC and Psycinfo databases on February 14, 2022. In addition, relevant publications were hand searched for relevant studies. We considered interventions that provided data on CB prevalence changes in populations between primary school and college age. Results: In total, 111 studies were retained for further screening from 3,477 results. Following rigorous screening, 43 reports including 46 studies and information from 36 different interventions were included in our systematic review. Results shows that most of the interventions measuring reductions in global CB, cyberperpetration/victimization, cybervictimization and cyberperpetration were effective or partially effective. While the interventions measuring reductions in cyber-bystanding were not effective. Multicomponent interventions showed higher effectiveness than single-component interventions. After completion of the TIDieR check-list, included interventions were considered to offer an insufficient level of detail for a number of the analyzed items in relation to "how well planned," "intervention modifications" and "tailoring." Conclusion: Given the aforementioned, it is critical to increase the number of studies and the quality of interventions targeting CB and the level of detail of its description in order to obtain more robust outcomes about how to reduce its prevalence and facilitate the replication of the effective interventions. Systematic review registration: https://archive.org/details/osf-registrations-wn5u4-v1, Identifer DOI: 10.17605/OSF.IO/WN5U4.

Single-cell analysis identifies genes facilitating rhizobium infection in Lotus japonicus.

Legume-rhizobium signaling during establishment of symbiotic nitrogen fixation restricts rhizobium colonization to specific cells. A limited number of root hair cells allow infection threads to form, and only a fraction of the epidermal infection threads progress to cortical layers to establish functional nodules. Here we use single-cell analysis to define the epidermal and cortical cell populations that respond to and facilitate rhizobium infection. We then identify high-confidence nodulation gene candidates based on their specific expression in these populations, pinpointing genes stably associated with infection across genotypes and time points. We show that one of these, which we name SYMRKL1, encodes a protein with an ectodomain predicted to be nearly identical to that of SYMRK and is required for normal infection thread formation. Our work disentangles cellular processes and transcriptional modules that were previously confounded due to lack of cellular resolution, providing a more detailed understanding of symbiotic interactions.

Using Hyper as a molecular probe to visualize hydrogen peroxide in living plant cells: An updated method.

Reactive oxygen species (ROS) are highly reactive reduced oxygen molecules that play a myriad of roles in animal and plant cells. In plant cells the production of ROS results from aerobic metabolism during respiration and photosynthesis. Therefore mitochondria, chloroplasts, and peroxisomes constitute an important source of ROS. However, ROS can also be produced in response to many physiological stimuli such as pathogen attack, hormone signaling, abiotic stresses or during cell wall organization and plant morphogenesis. The study of ROS in plant cells has been limited to biochemical assays and use of fluorescent probes, however, the irreversible oxidation of the fluorescent dyes prevents the visualization of dynamic changes. We have previously reported that Hyper 1 is a biosensor for H2O2 and consists of a circularly permutated YFP (cpYFP) inserted into the regulatory domain of the Escherichia coli hydrogen peroxide (H2O2) sensor protein OxyR rendering it an H2O2-specific quantitative probe (Bilan & Belousov, 2018; Hernandez-Barrera et al., 2015). Herein we describe an updated protocol for using the improved new version of Hyper 2 and Hyper 3 as a dynamic biosensor for H2O2 in Arabidopsis with virtually unlimited potential to detect H2O2 throughout the plant and under a broad range of developmental and environmental conditions (Bilan et al., 2013).

Correlation Between Opioid Drug Prescription and Opioid-Related Mortality in Spain as a Surveillance Tool: Ecological Study.

BACKGROUND: Opioid drug prescription (ODP) and opioid-related mortality (ORM) have increased in Spain. However, their relationship is complex, as ORM is registered without considering the type of opioid (legal or illegal). OBJECTIVE: This ecological study aimed to examine the correlation between ODP and ORM in Spain and discuss their usefulness as a surveillance tool. METHODS: This was an ecological descriptive study using retrospective annual data (2000-2019) from the Spanish general population. Data were collected from people of all ages. Information on ODP was obtained from the Spanish Medicines Agency in daily doses per 1000 inhabitants per day (DHD) for total ODP, total ODP excluding those with better safety protocols (codeine and tramadol), and each opioid drug separately. Rates of ORM (per 1,000,000 inhabitants) were calculated based on deaths registered (International Classification of Diseases, 10th Revision codes) as opioid poisoning by the National Statistics Institute, derived from the drug data recorded by medical examiners in death certificates. Opioid-related deaths were considered to be those that indicated opioid consumption (accidental, infringed, or self-inflicted) as the main cause of death: death due to accidental poisoning (X40-X44), intentional self-inflicted poisoning (X60-X64), drug-induced aggression (X85), and poisoning of undetermined intention (Y10-Y14). A descriptive analysis was carried out, and correlations between the annual rates of ORM and DHD of the prescribed opioid drugs globally, excluding medications of the least potential risk of overdose and lowest treatment tier, were analyzed using Pearson linear correlation coefficient. Their temporal evolution was analyzed using cross-correlations with 24 lags and the cross-correlation function. The analyses were carried out using Stata and StatGraphics Centurion 19. RESULTS: The rate of ORM (2000-2019) ranged between 14 and 23 deaths per 1,000,000 inhabitants, with a minimum in 2006 and an increasing trend starting in 2010. The ODP ranged between 1.51 to 19.94 DHD. The rates of ORM were directly correlated with the DHD of total ODP (r=0.597; P=.006), total ODP without codeine and tramadol (r=0.934; P<.001), and every prescribed opioid except buprenorphine (P=.47). In the time analysis, correlations between DHD and ORM were observed in the same year, although not statistically significant (all P≥.05). CONCLUSIONS: There is a correlation between greater availability of prescribed opioid drugs and an increase in opioid-related deaths. The correlation between ODP and ORM may be a useful tool in monitoring legal opiates and possible disturbances in the illegal market. The role of tramadol (an easily prescribed opioid) is important in this correlation, as is that of fentanyl (the strongest opioid). Measures stronger than recommendations need to be taken to reduce off-label prescribing. This study shows that not only is opioid use directly related to the prescribing of opioid drugs above what is desirable but also an increase in deaths.